Biochemistry 2025

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Q.1 Subject and tasks of biological chemistry.

Ans. The subject of biological chemistry, also known as biochemistry, is the science that studies the chemical composition of
living matter, the functions of biomolecules, and the chemical transformations that underlie the vital functions of organisms. It
explores how the properties of living organisms arise from interactions among thousands of lifeless biomolecules, governed by
physical and chemical laws applicable to nonliving matter. Biochemistry aims to understand the molecular basis of life by
describing the structures, mechanisms, and chemical processes shared by all organisms, thereby revealing the molecular logic
of life1.
Subject of Biological Chemistry
• Study of chemical composition of living organisms, including plants, bacteria, viruses, animals, and humans.
• Examination of biomolecules such as proteins, lipids, carbohydrates, nucleic acids, and complex molecules.
• Understanding molecular interactions that maintain life processes.
• Analysis of metabolic pathways, energy transformations, and molecular mechanisms of cellular functions.
• Investigation of molecular basis of heredity, enzyme action, and biochemical regulation.
• Exploration of biochemical processes in health, disease, and therapy1234.
Main Tasks of Biological Chemistry
 1. Study Chemical Composition of the Human Body and Other Organisms
Investigate the molecular components and their structures, such as amino acids, proteins, carbohydrates, lipids, nucleic
acids, and complex conjugated proteins124.
2. Establish Structure-Function Relationships
Analyze how the structure of biomolecules determines their specific functions in biological systems, including enzymes,
transport proteins, and regulatory molecules124.
3. Understand Metabolic Processes and Biotransformation
Study the chemical reactions and pathways (catabolism and anabolism) that ensure exchange of substances, energy, and
information between the body and environment, including digestion, respiration, and biosynthesis135.
4. Assess Metabolic Plasticity and Renewal
Investigate the dynamic metabolic functions that underlie structural renewal and adaptation in cells and tissues1.
5. Explore Molecular Basis of Cellular Self-Reproduction and Information Transfer
Understand how genetic information encoded in DNA is transformed into proteins and how cells replicate and maintain
their functions14.
6. Develop Knowledge for Rational Therapy and Disease Prevention
Apply biochemical principles to medicine, nutrition, and pharmacology to improve health, develop diagnostics, and
design therapies169.

7. Promote Understanding of Unity and Diversity of Life

Consider the biochemical unity of living nature and the human being as part of the biosphere, reflecting evolutionary
relationships1.

2. Amino acids, chemical structure, properties. Types of isomerism.

Ans. Amino acids are the fundamental building blocks of proteins and play crucial roles in metabolism, signaling, and structure
within living organisms.
Chemical Structure of Amino Acids
• All standard amino acids share a common structure:
• A central (alpha, αα) carbon atom
• An amino group (−NH2−NH2)
• A carboxyl group (−COOH−COOH)
• A hydrogen atom
• A unique side chain (R group) that differs for each amino acid
This general structure is represented as:
H2N−CH(R)−COOHH2N−CH(R)−COOH
• The αα-carbon is typically a chiral center (except in glycine, where R = H), meaning it is attached to four different
groups2.
Properties of Amino Acids
• Amphoteric Nature: Amino acids can act as both acids and bases due to the presence of both amino and carboxyl
groups. In aqueous solutions at physiological pH, amino acids exist predominantly as zwitterions, with a positively
charged amino group (−NH3+−NH3+) and a negatively charged carboxyl group (−COO−−COO−)2.
• Solubility: The solubility of amino acids in water depends on the nature of the R group. Nonpolar R groups make amino
acids hydrophobic, while polar or charged R groups make them hydrophilic2.
• Classification by R group: Amino acids are grouped based on the properties of their side chains:
• Nonpolar, aliphatic (e.g., glycine, alanine, valine)
 • Aromatic (e.g., phenylalanine, tyrosine, tryptophan)
• Polar, uncharged (e.g., serine, threonine)
• Positively charged (basic; e.g., lysine, arginine)
• Negatively charged (acidic; e.g., aspartate, glutamate)2.
• Essential vs. Non-Essential: Some amino acids cannot be synthesized by the human body and must be obtained from
the diet (essential amino acids)2.
Types of Isomerism in Amino Acids
1. Stereoisomerism (Optical Isomerism):
• Most amino acids (except glycine) have a chiral αα-carbon and exist as two enantiomers: L- and D-forms.
• These are non-superimposable mirror images. In proteins, only the L-form is found naturally2.
• The D/L system is based on the configuration relative to glyceraldehyde.
2. Structural Isomerism:
• While all standard amino acids are αα-amino acids, some non-standard amino acids can have the amino group attached
to a different carbon (e.g., ββ- or γγ-amino acids), but these are rare in proteins.
3. Geometric Isomerism:
• Not typical for standard amino acids, but relevant in the context of peptide bonds (cis/trans isomerism) and in certain
modified amino acids.

3. Classification of amino acids by radical and degree of irreplaceability.

Ans. Classification by Radical (Side Chain) Properties
Amino acids are most commonly classified based on the chemical nature of their side chains (R groups), which profoundly
influence their chemical behavior and biological roles. The main classes are1:
• Nonpolar, Aliphatic R Groups:
• Hydrophobic, tend to avoid water.
• Examples: Glycine, Alanine, Valine, Leucine, Isoleucine, Methionine, Proline.
• Aromatic R Groups:
• Contain aromatic rings, hydrophobic but can participate in specific interactions.
• Examples: Phenylalanine, Tyrosine, Tryptophan.
• Polar, Uncharged R Groups:
• Hydrophilic, can form hydrogen bonds.
• Examples: Serine, Threonine, Cysteine, Asparagine, Glutamine.
• Positively Charged (Basic) R Groups:
• Hydrophilic, carry a positive charge at physiological pH.
• Examples: Lysine, Arginine, Histidine.
• Negatively Charged (Acidic) R Groups:
• Hydrophilic, carry a negative charge at physiological pH.
• Examples: Aspartate (Aspartic acid), Glutamate (Glutamic acid).
This classification reflects the amino acid’s polarity, charge, and ability to interact with water or other molecules, which is
central to protein structure and function1.
Classification by Degree of Irreplaceability (Essentiality)
Amino acids are also classified based on whether they can be synthesized by the human body or must be obtained from the
diet1:

4. Simple proteins, chemical structure. The levels of organization of a protein molecule.

Ans. Simple Proteins: Chemical Structure
Simple proteins are macromolecules composed exclusively of amino acids, without any non-protein (prosthetic) groups
attached. Their structure is determined by the sequence and composition of 20 standard amino acids, which are covalently
linked together by peptide bonds in a linear chain. Each peptide bond forms between the carboxyl group of one amino acid
and the amino group of the next, releasing a molecule of water (a dehydration reaction). When many amino acids are joined in
this way, the resulting polymer is called a polypeptide; if the chain is long enough and folds into a functional three-dimensional
shape, it is considered a protein1.
The chemical structure of a simple protein is thus a specific sequence of amino acid residues, each with a central alpha-carbon
bonded to an amino group, a carboxyl group, a hydrogen atom, and a unique side chain (R group). The side chains determine
the chemical properties and biological functions of the protein1.
Levels of Organization of a Protein Molecule
Protein structure is organized into four hierarchical levels:
1. Primary Structure
This is the unique linear sequence of amino acids in the polypeptide chain, held together by covalent peptide bonds. The
primary structure determines all higher levels of protein structure and function1.
2. Secondary Structure
This refers to localized, regular folding patterns within the polypeptide chain, stabilized primarily by hydrogen bonds between
backbone atoms. The most common secondary structures are the alpha-helix and beta-pleated sheet. These structures do not
involve side chain interactions, but rather the backbone of the polypeptide chain1.
3. Tertiary Structure
The tertiary structure is the overall three-dimensional shape of a single polypeptide chain. It results from interactions between
the side chains (R groups) of amino acids, including hydrogen bonds, ionic bonds, hydrophobic interactions, and disulfide
bridges. This level of structure determines the protein's functional conformation and is essential for its biological activity1.
4. Quaternary Structure
Some proteins consist of more than one polypeptide chain (subunit). The quaternary structure describes how these individual
subunits associate and interact to form a functional protein complex. The interactions are similar to those stabilizing tertiary
structure but occur between separate polypeptide chains1.
Summary:
Simple proteins are polymers of amino acids linked by peptide bonds, with their structure organized into primary (sequence),
secondary (local folding), tertiary (overall 3D conformation), and quaternary (subunit assembly) levels. Each level of
organization is critical for the protein's stability and function1.

5. Classification of simple proteins, characteristics of individual classes.

Ans. Simple proteins are those that yield only amino acids upon hydrolysis and do not contain any non-protein (prosthetic)
groups. They are composed exclusively of amino acid residues linked by peptide bonds and represent the most basic form of
proteins in living organisms.
Classification of Simple Proteins
Simple proteins are classified based on their solubility, physical properties, and biological functions. The main classes and their
characteristics are:
1. Albumins
• Solubility: Soluble in water and dilute salt solutions.
• Properties: Coagulate upon heating; have a relatively low molecular weight.
• Biological Role: Maintain osmotic pressure in blood (e.g., serum albumin), serve as transport proteins.
• Examples: Serum albumin, egg albumin (ovalbumin), lactalbumin1.
2. Globulins
• Solubility: Insoluble in pure water but soluble in dilute salt solutions.
• Properties: Coagulate by heat; higher molecular weight than albumins.
• Biological Role: Play roles in immune defense (immunoglobulins/antibodies), transport, and storage.
• Examples: Serum globulins, ovoglobulin, legumin (plant globulin)1.
3. Glutelins
• Solubility: Insoluble in water and neutral salt solutions; soluble in dilute acids or bases.
• Properties: Found mainly in plant seeds.
• Biological Role: Serve as storage proteins in plants.
• Examples: Glutenin (wheat), oryzenin (rice)1.
4. Prolamins
• Solubility: Insoluble in water and neutral salt solutions; soluble in 70–80% ethanol.
 • Properties: Rich in proline and glutamine.
• Biological Role: Storage proteins in cereal grains.
• Examples: Gliadin (wheat), zein (corn), hordein (barley)1.
5. Histones
• Solubility: Soluble in water; basic in nature due to high content of lysine and arginine.
• Properties: Do not coagulate by heat.
• Biological Role: Bind to DNA and play a structural role in chromatin organization in cell nuclei.
• Examples: Various histone types (H1, H2A, H2B, H3, H4)2.
6. Protamines
• Solubility: Soluble in water; strongly basic due to high arginine content.
• Properties: Do not coagulate by heat; have a very low molecular weight.
• Biological Role: Combine with nucleic acids, especially in fish sperm.
• Examples: Salmine (from salmon), sturine (from sturgeon)2.
Summary of Key Features
• Albumins and globulins are the most abundant simple proteins in animal tissues and fluids, with major roles in
transport, immunity, and maintaining osmotic balance.
• Glutelins and prolamins are primarily plant storage proteins, important in human nutrition and food science.
• Histones and protamines are basic proteins associated with nucleic acids, essential for DNA packaging and regulation in
cells.
This classification is fundamental in biochemistry and underpins the understanding of protein function, nutrition, and
molecular biology1.
6. Biological functions of proteins.
Ans. functions necessary for life. Their diverse roles are determined by their unique structures and the specific sequence of
amino acids in each protein molecule. The main biological functions of proteins include:
1. Catalytic Function
• Enzymes are proteins that act as biological catalysts, accelerating the rate of biochemical reactions without being
consumed in the process. Nearly all enzymes are proteins, and they are crucial for metabolism, DNA replication, energy
production, and many other cellular processes73.
2. Structural Function
• Proteins provide structural support and shape to cells and tissues. Examples include collagen in connective
tissues, keratin in hair and nails, and elastin in ligaments and skin. These proteins form the framework that maintains
the integrity and resilience of biological structures23.
3. Transport Function
• Many proteins are involved in the transport of molecules within organisms. Hemoglobin transports oxygen in the
blood, albumin carries hormones and drugs, and membrane transport proteins facilitate the movement of ions and
nutrients across cell membranes123.
4. Regulatory Function
• Proteins act as regulators of biological processes. This includes hormones (such as insulin and growth hormone), which
control metabolism, growth, and development, and transcription factors, which regulate gene expression23.
5. Protective Function
 • Some proteins protect the body from pathogens and injury. Antibodies (immunoglobulins) are proteins that identify
and neutralize foreign invaders like bacteria and viruses. Fibrinogen and thrombin are involved in blood clotting,
preventing excessive bleeding23.
6. Contractile Function
• Contractile proteins are responsible for movement. Actin and myosin in muscle fibers enable muscle contraction, while
similar proteins are involved in cell division and intracellular transport23.
7. Energy Function
• Proteins can serve as an energy source, especially when carbohydrates and fats are insufficient. During prolonged fasting
or starvation, proteins are broken down to provide energy for vital processes23.
8. Receptor and Signal Transduction Function
• Proteins function as receptors on cell surfaces, detecting and responding to chemical signals such as hormones,
neurotransmitters, and growth factors, thereby mediating communication between cells and their environment3.
9. Storage Function
• Some proteins store essential substances for later use. For example, ferritin stores iron in the liver, and casein stores
amino acids in milk for the nourishment of offspring13.
10. Participation in Genetic Information Transfer
• Proteins are involved in the storage, replication, and transmission of genetic information. Histones package DNA in the
nucleus, and various proteins participate in DNA replication, repair, and transcription13.

7. Physicochemical properties of proteins. Difference in molecular shape, molecular weight, total charge, solubility. Factors of
stability of a protein molecule in solution.
Ans. Physicochemical Properties of Proteins
Molecular Shape
Proteins exhibit two principal molecular shapes: fibrous and globular.
• Fibrous proteins are elongated, strand-like molecules, often serving structural roles (e.g., collagen, keratin). They are
generally insoluble in water and form tough, protective structures.
• Globular proteins are compact, roughly spherical, and typically soluble in water. They perform dynamic functions such
as catalysis (enzymes), transport (hemoglobin), and regulation (hormones)1.
This difference in shape is a direct result of the amino acid sequence and the way the polypeptide chain folds into secondary,
tertiary, and sometimes quaternary structures1.
Molecular Weight
Proteins are polymers of amino acids and can vary greatly in size:
• Polypeptides with molecular weights below 10,000 Da are generally called peptides.
• Proteins have higher molecular weights, often ranging from several thousand to several million Daltons, depending on
the number and type of amino acids incorporated1.
The molecular weight affects the protein’s physical properties, such as diffusion, sedimentation, and its behavior during
purification.
Total Charge
The total charge of a protein molecule depends on the pH of its environment and the nature of its amino acid side chains:
• At acidic pH, proteins carry a net positive charge.
• At alkaline pH, they carry a net negative charge.
• The isoelectric point (pI) is the pH at which the net charge is zero. At this point, proteins are least soluble and may
precipitate1.
 • The charge influences protein solubility, hydration, and stability in solution.
Solubility
Protein solubility is determined by several factors:
• Shape: Globular proteins are generally more soluble in water than fibrous proteins.
• Charge: Solubility is lowest at the isoelectric point and increases as the net charge (positive or negative) increases.
• R group properties: Amino acids with polar or charged side chains increase solubility, while nonpolar side chains
decrease it.
• Environmental conditions: pH, salt concentration, temperature, and the presence of solvents or detergents all affect
solubility1.
Factors of Stability of a Protein Molecule in Solution
Several factors contribute to the stability of proteins in solution:
• Hydration: Water molecules form a solvation shell around the protein, stabilizing it and preventing aggregation or
precipitation.
• pH: Stability is highest away from the isoelectric point; extreme pH can lead to denaturation.
• Ionic Strength: Moderate salt concentrations can stabilize proteins by shielding charges, but high concentrations can
cause "salting out" and precipitation.
• Temperature: High temperatures can disrupt non-covalent interactions, leading to denaturation.
• Presence of stabilizing/destabilizing agents: Organic solvents, heavy metals, detergents, and other chemicals can
destabilize proteins by disrupting hydrophobic, ionic, or hydrogen bonds1.

8. Protein denaturation, mechanism and factors causing protein denaturation. Meaning. Examples. Medical applications.
Ans. Protein Denaturation: Meaning, Mechanism, Factors, Examples, and Medical Applications
Meaning of Protein Denaturation
Protein denaturation is the process by which a protein loses its native three-dimensional structure-secondary, tertiary, and
quaternary organization-without the breaking of peptide (primary structure) bonds. This structural disruption leads to the loss
of the protein’s biological activity and function. If the primary structure is also destroyed, the process is called hydrolysis rather
than denaturation3.
Mechanism of Denaturation
Denaturation involves the disruption of the non-covalent interactions that maintain a protein’s higher-order structure:
• Hydrogen bonds (between backbone and side chains)
• Ionic bonds (between charged side chains)
• Hydrophobic interactions (among nonpolar side chains)
• Disulfide bridges (covalent, but can be disrupted by reducing agents)
When these interactions are disturbed, the protein unfolds or aggregates, losing its specific shape and, consequently, its
function. Denaturation does not break the peptide bonds that form the primary structure3.
Factors Causing Protein Denaturation
Physical Factors:
• Heat: Increases molecular motion, disrupting hydrogen bonds and hydrophobic interactions.
• Ultrasound: Mechanical agitation can disrupt protein structure.
• Ionizing radiation: Can break bonds and alter side chains.
Chemical Factors:
 • Acids and alkalis: Alter charge distribution, disrupt ionic bonds and hydrogen bonding.
• Organic solvents: (e.g., alcohol, acetone) Disrupt hydrophobic interactions.
• Heavy metal salts: Interact with side chains, especially sulfur-containing amino acids, leading to precipitation.
• Detergents: Disrupt hydrophobic interactions and solubilize proteins3.
Examples of Protein Denaturation
• Cooking an egg: Heat denatures ovalbumin in egg white, turning it from transparent to opaque and solid.
• Sterilization: Heat or chemicals denature microbial proteins, killing pathogens.
• Alcohol disinfection: Alcohol denatures proteins in bacterial cell walls and membranes.
• Milk curdling: Acid or enzymes denature casein, leading to coagulation.
Medical Applications of Protein Denaturation
• Disinfection and Sterilization: Alcohols, heat, and other agents denature microbial proteins, making them essential for
sterilizing medical instruments and surfaces.
• Diagnostic Tests: Protein denaturation is used in clinical tests, such as the heat coagulation test for protein in urine
(diagnosing kidney disease).
• Vaccine Production: Toxoids are vaccines made from denatured bacterial toxins (e.g., tetanus, diphtheria), which lose
toxicity but retain immunogenicity.
• Enzyme Inactivation: Controlled denaturation is used to inactivate enzymes during certain therapeutic or laboratory
procedures3.
Renaturation
If the denaturing agent is removed and the primary structure remains intact, some proteins can refold into their native
conformation and regain function-a process called renaturation.
9. Protein separation and purification methods: salting out, dialysis, electrophoresis, chromatography. Types, principle,
application.
Ans. Protein Separation and Purification Methods
Protein separation and purification are essential steps in biochemical research and clinical diagnostics. The main methods
include salting out, dialysis, electrophoresis, and chromatography. Each exploits different physicochemical properties of
proteins to achieve separation and purification.
Salting Out
Principle:
Salting out is based on the principle that protein solubility decreases at high salt concentrations. As salt is added, water
molecules preferentially interact with the salt ions, reducing the amount of water available to solubilize proteins. This causes
proteins to aggregate and precipitate out of solution.
Types:
The most common salt used is ammonium sulfate due to its high solubility and gentle effect on protein structure.
Application:
• Used as an initial step to fractionate proteins from a crude extract.
• Different proteins precipitate at different salt concentrations, allowing for selective separation12.
Dialysis
Principle:
Dialysis separates proteins from small molecules (such as salts, solvents, or metabolites) using a semipermeable membrane.
The protein solution is placed inside a dialysis bag or tube, which is then immersed in a large volume of buffer. Small molecules
diffuse out through the membrane, while proteins are retained due to their larger size.
Types:
• Standard dialysis (removal of salts and small molecules)
• Ultrafiltration (uses membranes with different molecular weight cut-offs for more selective separation)
Application:
• Removal of salts after salting out or precipitation steps.
• Buffer exchange for further purification or analysis12.
Electrophoresis
Principle:
Electrophoresis separates proteins based on their size and charge by applying an electric field to a gel matrix (commonly
polyacrylamide). Proteins migrate through the gel at rates determined by their net charge and molecular weight.
Types:
• SDS-PAGE (Sodium Dodecyl Sulfate Polyacrylamide Gel Electrophoresis): Denatures proteins and imparts a uniform
negative charge, separating them by size.
• Native PAGE: Separates proteins in their native, non-denatured state, based on both size and charge.
• Isoelectric focusing: Separates proteins based on their isoelectric point (pI).
Application:
• Analysis of protein purity and molecular weight.
• Identification and characterization of proteins.
• Clinical diagnostics (e.g., detection of abnormal hemoglobins or serum proteins)1.
Chromatography
Principle:
Chromatography separates proteins based on differences in their physical or chemical properties as they pass through a
stationary phase (solid matrix) and a mobile phase (liquid buffer).
Types and Their Principles:
• Ion-Exchange Chromatography: Separates proteins by charge. The stationary phase contains charged groups that
interact with oppositely charged protein residues. Proteins are eluted by changing the pH or salt concentration.
• Size-Exclusion (Gel Filtration) Chromatography: Separates proteins by size. The stationary phase consists of porous
beads; larger proteins are excluded from the pores and elute first, while smaller proteins enter the pores and elute later.
• Affinity Chromatography: Separates proteins by specific binding to a ligand attached to the stationary phase. Only
proteins with affinity for the ligand are retained and later eluted by adding a competing ligand or changing the buffer
conditions.
Application:
• Ion-exchange: Purification of proteins with different net charges.
• Size-exclusion: Desalting and separation of proteins from aggregates or contaminants.
• Affinity: Highly specific purification, such as isolating antibodies, enzymes, or tagged proteins12.
Summary of Applications
• Salting out: Crude fractionation of proteins from cell extracts.
• Dialysis: Removal of small molecules and buffer exchange.
• Electrophoresis: Analytical separation for purity and molecular weight assessment.
• Chromatography: High-resolution purification and isolation of specific proteins for research, diagnostics, or therapeutic
use.
Each method can be used alone or in combination, depending on the complexity of the protein mixture and the required
purity level.

10. Complex proteins, chemical structure, classification.

Ans. Chemical Structure of Complex Proteins
Complex proteins, also known as conjugated proteins, are macromolecules composed of a simple protein portion (apoprotein)
covalently or non-covalently bound to a non-protein component called the prosthetic group. The complete, functional
molecule is called a holoprotein. Upon hydrolysis, complex proteins yield both amino acids and their specific non-protein
prosthetic groups13.
• Apoprotein: The protein part, built from amino acid residues linked by peptide bonds.
• Prosthetic Group: The non-protein component, which can be organic (e.g., heme, carbohydrate, lipid) or inorganic (e.g.,
metal ion). The prosthetic group is essential for the biological function of the protein13.
The nature of the prosthetic group determines the classification and function of the complex protein.
Classification of Complex Proteins
Complex proteins are classified based on the type of prosthetic group attached to the apoprotein. The main classes include13:
1. Nucleoproteins
• Prosthetic group: Nucleic acids (DNA or RNA)
• Function: Storage, replication, recombination, and transmission of genetic information. Examples include chromatin
(DNA + histones) and ribosomes (RNA + proteins).
2. Glycoproteins
• Prosthetic group: Carbohydrates (monosaccharides or oligosaccharides)
 • Function: Structural (collagens), protective (mucins), transport (transferrin), immune response (immunoglobulins),
hormones (thyroid-stimulating hormone), enzymes, cell recognition, and adhesion. Carbohydrates are attached via O-
linked (Ser, Thr) or N-linked (Asn) glycosidic bonds.
3. Proteoglycans
• Prosthetic group: Glycosaminoglycans (large, negatively charged polysaccharides)
• Function: Provide viscosity, adhesiveness, and tensile strength to the extracellular matrix (e.g., cartilage, connective
tissue).
4. Lipoproteins
• Prosthetic group: Lipids (triacylglycerols, phospholipids, cholesterol)
• Function: Transport of lipids in blood (e.g., HDL, LDL, VLDL, chylomicrons), structural roles in membranes.
5. Phosphoproteins
• Prosthetic group: Phosphate groups (usually on Ser, Thr, or Tyr residues)
• Function: Regulation (via reversible phosphorylation), nutrition (casein in milk), structural and catalytic roles.
6. Metalloproteins
• Prosthetic group: Metal ions (e.g., Fe, Zn, Cu, Ca, Mo)
• Function: Transport (hemoglobin, transferrin), catalysis (carbonic anhydrase, ceruloplasmin), structural (ferritin), and
regulatory functions.
7. Hemoproteins
• Prosthetic group: Heme (iron-containing porphyrin ring)
• Function: Oxygen transport (hemoglobin, myoglobin), catalysis (cytochromes, catalase, peroxidase), electron transfer.
8. Flavoproteins
• Prosthetic group: Flavin nucleotides (FMN, FAD)
• Function: Redox reactions in metabolism (e.g., succinate dehydrogenase, cytochrome P450 reductase).
9. Chromoproteins
• Prosthetic group: Pigmented groups (e.g., heme, flavin)
• Function: Transport, catalysis, energy (hemoglobin, myoglobin, cytochromes, phytochromes)
11. Nucleoproteins, chemical structure, functions.
Ans. Nucleoproteins: Chemical Structure and Functions
Chemical Structure
Nucleoproteins are a class of complex (conjugated) proteins composed of two main components:
• Protein part (apoprotein): Typically, this is a basic protein such as a histone or protamine.
• Nucleic acid (prosthetic group): Either DNA or RNA is tightly bound to the protein component.
Depending on the type of nucleic acid, nucleoproteins are classified as:
• Deoxyribonucleoproteins: Contain DNA (e.g., chromatin in cell nuclei).
• Ribonucleoproteins: Contain RNA (e.g., ribosomes, some viruses).
Structural Features:
• In chromatin, DNA is wrapped around histone proteins to form nucleosomes, which further organize into higher-order
structures, ultimately forming chromosomes.
• Histones are rich in basic amino acids (lysine, arginine, histidine), which facilitate tight binding to the negatively charged
phosphate backbone of nucleic acids.
 • In ribonucleoproteins, proteins associate with RNA to form functional complexes, such as ribosomes (the site of protein
synthesis) and spliceosomes (involved in RNA processing)25.
Functions of Nucleoproteins
Nucleoproteins play fundamental biological roles, primarily related to the storage, expression, and transmission of genetic
information:
1. Storage of Genetic Information
• In the form of chromatin, nucleoproteins compact and organize DNA within the cell nucleus, making efficient use of
space and protecting genetic material from damage2.
2. Replication of Genetic Material
• Nucleoproteins are essential for the accurate duplication of DNA during cell division. Histones and other chromatin-
associated proteins regulate access to DNA and participate in the replication process2.
3. Recombination
• Nucleoproteins facilitate genetic recombination, which is crucial for genetic diversity and repair of DNA damage2.
4. Transmission of Genetic Information
• Nucleoproteins ensure the faithful transmission of genetic information from one generation to the next during cell
division and reproduction25.
5. Regulation of Gene Expression
• By modifying chromatin structure (e.g., through histone modification), nucleoproteins regulate which genes are
accessible for transcription, thus controlling gene expression patterns2.
6. Protein Synthesis
 • Ribonucleoproteins, such as ribosomes, are essential for translating messenger RNA (mRNA) into proteins, a central
process in gene expression25.
7. RNA Processing and Transport
• Ribonucleoproteins are involved in the processing of precursor RNA molecules (splicing, modification) and their
transport within the cell2.

12. The structure of nucleic acids. Primary, secondary, tertiary, the difference in the structure of DNA and RNA.
Ans. Structure of Nucleic Acids: Primary, Secondary, Tertiary; Differences Between DNA and RNA
Primary Structure
The primary structure of nucleic acids is the linear sequence of nucleotides linked by phosphodiester bonds. Each nucleotide
consists of:
• A pentose sugar (deoxyribose in DNA, ribose in RNA)
• A phosphate group
• A nitrogenous base (purines: adenine [A], guanine [G]; pyrimidines: cytosine [C], thymine [T] in DNA, uracil [U] in RNA)
The sequence of these nucleotides encodes genetic information. The backbone is formed by alternating sugar and phosphate
groups, with bases projecting from the sugar2.
Secondary Structure
The secondary structure refers to the local folding and base pairing within the nucleic acid:
• DNA: The classic form is the double helix, where two antiparallel strands are held together by hydrogen bonds between
complementary bases (A-T, G-C). The double helix is right-handed (B-form) and stabilized by base stacking interactions
and hydrogen bonds.
 • RNA: Usually single-stranded but can form regions of double helix by folding back on itself, creating structures like
hairpins, loops, and stems. Base pairing is A-U and G-C in RNA2.
Tertiary Structure
Tertiary structure is the three-dimensional arrangement of the nucleic acid molecule:
• DNA: Supercoiling and higher-order folding (as seen in chromosomes, where DNA wraps around histone proteins to
form nucleosomes and further compacts into chromatin).
• RNA: Complex three-dimensional shapes due to extensive intramolecular base pairing and folding, crucial for its
function (e.g., tRNA, rRNA)2.
Differences in Structure: DNA vs. RNA
1. Strandedness and Length
• DNA: Double-stranded, forms a long, stable double helix; much longer molecules (chromosomes can be several
centimeters long when stretched out).
• RNA: Usually single-stranded, shorter molecules; can form local double-helical regions but rarely a full double helix2.
2. Sugar Component
• DNA: Contains deoxyribose (lacks an oxygen atom at the 2' position).
• RNA: Contains ribose (has a hydroxyl group at the 2' position), making RNA more reactive and less stable than DNA2.
3. Nitrogenous Bases
• DNA: Adenine (A), Guanine (G), Cytosine (C), Thymine (T).
• RNA: Adenine (A), Guanine (G), Cytosine (C), Uracil (U) replaces thymine2.
4. Base Pairing
 • DNA: A pairs with T (two hydrogen bonds), G pairs with C (three hydrogen bonds).
• RNA: A pairs with U, G pairs with C; base pairing occurs in local regions due to folding2.
5. Location and Function
• DNA: Located primarily in the nucleus (and mitochondria), serves as the long-term storage of genetic information.
• RNA: Synthesized in the nucleus but functions mainly in the cytoplasm; acts as a messenger (mRNA), adapter (tRNA), or
structural/catalytic component (rRNA)2.
6. Stability
• DNA: More chemically stable due to the absence of the 2' hydroxyl group and double-stranded structure.
• RNA: Less stable, more easily degraded, and more reactive due to the 2' hydroxyl group2.

13. Hemoproteins, chemical structure, representatives, biological role.

Ans. Chemical Structure
Hemoproteins are complex (conjugated) proteins that contain a heme prosthetic group tightly bound to their polypeptide
(apoprotein) component. The heme group is a planar, pigmented structure composed of:
• Protoporphyrin IX ring: a large, cyclic organic molecule made of four pyrrole rings linked by methene bridges.
• Central iron atom (Fe²⁺): coordinated to the four nitrogens of the porphyrin ring and capable of binding additional
ligands above and below the plane.
The iron atom in heme is essential for the protein’s function, as it can reversibly bind small molecules such as oxygen, carbon
monoxide, or participate in redox reactions. The protein part (globin or enzyme) creates a specific environment around the
heme, modulating its reactivity and specificity1.
Representatives
Major classes of hemoproteins include:
• Hemoglobin:
• Found in red blood cells.
• Tetrameric structure (four subunits, each with its own heme group).
• Transports oxygen from the lungs to tissues and facilitates carbon dioxide transport back to the lungs.
• Variants include adult hemoglobin (HbA), fetal hemoglobin (HbF), and abnormal forms (HbS in sickle cell disease,
HbC, etc.)1.
• Myoglobin:
• Present in cardiac and skeletal muscle.
• Monomeric protein (single polypeptide with one heme group).
• Stores and facilitates the diffusion of oxygen within muscle tissue1.
• Cytochromes:
• A family of hemoproteins involved in electron transport and redox reactions.
• Key components of the mitochondrial electron transport chain (e.g., cytochrome c, cytochrome b, cytochrome
a/a3).
• Participate in cellular respiration and energy production13.
• Catalase and Peroxidase:
• Enzymes that contain heme and catalyze the breakdown of hydrogen peroxide (H₂O₂) and organic peroxides,
protecting cells from oxidative damage1.
• Other Hemoproteins:
 • Cytochrome P450 enzymes: involved in drug metabolism and biosynthesis of steroid hormones.
• Various other oxidases and peroxidases13.
Biological Role
1. Oxygen Transport and Storage
• Hemoglobin binds oxygen in the lungs and releases it in tissues; also acts as a buffer and antioxidant, and participates in
iron metabolism regulation.
• Myoglobin stores oxygen in muscles, releasing it during periods of high demand or low oxygen availability1.
2. Electron Transport and Energy Production
• Cytochromes are critical for electron transfer in the mitochondrial respiratory chain, enabling ATP synthesis through
oxidative phosphorylation3.
3. Catalysis of Redox Reactions
• Catalase and peroxidase enzymes decompose hydrogen peroxide and organic peroxides, preventing oxidative damage
and supporting cellular detoxification13.
4. Metabolism and Detoxification
• Cytochrome P450 enzymes in the liver metabolize drugs, toxins, and endogenous compounds by catalyzing oxidation
reactions3.
5. Other Roles
• Hemoproteins play roles in signal transduction, cellular defense, and regulation of various metabolic pathways.

14. Hemoglobin, chemical structure. Types and variants of hemoglobin.

Ans. Chemical Structure
Hemoglobin is a complex (conjugated) protein found primarily in red blood cells. Its structure consists of two main
components:
• Globin: The protein part, composed of four polypeptide chains (subunits). In adult hemoglobin (HbA), there are two
alpha (α) and two beta (β) chains. Each chain is arranged in a series of alpha-helix segments, forming a compact globular
structure.
• Heme: The non-protein prosthetic group, one per globin chain. Heme is a planar, pigmented molecule made of
protoporphyrin IX (a tetrapyrrole ring) with a central iron atom (Fe²⁺). The iron atom is coordinated to the four nitrogen
atoms of the porphyrin ring and can bind oxygen reversibly.
The hemoglobin molecule is a tetramer (α₂β₂ in adults), with each subunit containing its own heme group, allowing one
hemoglobin molecule to bind up to four oxygen molecules. The heme groups are located in hydrophobic pockets on the
surface of each globin subunit12.
Types and Variants of Hemoglobin
Hemoglobin exists in several forms throughout human development and in various genetic and acquired conditions:
Physiological Types
• Embryonic Hemoglobins:
• Gower 1 (ζ₂ε₂)
• Gower 2 (α₂ε₂)
• Portland I (ζ₂γ₂)
• Portland II (ζ₂β₂)
These are present in the early embryo and are replaced as development progresses.
• Fetal Hemoglobin (HbF):
 • Structure: α₂γ₂
• Predominant during fetal development; has a higher affinity for oxygen than adult forms. In adults, HbF is usually
restricted to a small subset of red cells (F-cells), but levels may be elevated in certain diseases (e.g., sickle cell
disease, β-thalassemia)1.
• Adult Hemoglobins:
• Hemoglobin A (HbA): α₂β₂ - the main form in healthy adults (>95%).
• Hemoglobin A₂ (HbA₂): α₂δ₂ - minor adult form (1.5–3.5%), with δ-chain synthesis beginning late in gestation1.
Pathological and Variant Forms
• Hemoglobin S (HbS):
• Structure: α₂βS₂ (mutation in β-chain)
• Found in sickle cell disease; causes red blood cells to assume a sickle shape under low oxygen, leading to anemia
and other complications.
• Hemoglobin C (HbC):
• Structure: α₂βC₂ (mutation in β-chain)
• Causes mild chronic hemolytic anemia; glutamic acid at position 6 of β-chain is replaced by lysine1.
• Hemoglobin D-Punjab:
• Structure: α₂βD₂
• Another variant due to a β-chain mutation.
• Hemoglobin H (HbH):
• Structure: β₄ (tetramer of β-chains)
 • Seen in some forms of α-thalassemia.
• Other Forms:
• Carboxyhemoglobin (COHb): Hemoglobin bound to carbon monoxide, preventing oxygen transport.
• Methemoglobin (MetHb): Iron in heme is oxidized to Fe³⁺, which cannot bind oxygen.
• Sulfhemoglobin: Hemoglobin irreversibly bound to sulfur, incapable of oxygen transport.
• Glycated Hemoglobin (HbA1c): Hemoglobin with glucose attached, used as a marker for long-term blood glucose
control in diabetes1.

15. Pathological types of hemoglobin, chemical structure. Sickle cell anemia, thalassemia.
Ans. Pathological Types of Hemoglobin and Their Chemical Structure
Pathological (abnormal) hemoglobins arise from mutations in the genes encoding the globin chains of hemoglobin. These
mutations typically result in single amino acid substitutions, deletions, or abnormal synthesis rates, leading to structural and
functional changes in the hemoglobin molecule. The most clinically significant pathological types include:
• Hemoglobin S (HbS):
• Chemical Structure: In HbS, the sixth amino acid in the β-globin chain is changed from glutamic acid (hydrophilic)
to valine (hydrophobic) due to a point mutation in the HBB gene. This creates a hydrophobic patch that causes
hemoglobin molecules to aggregate and form fibers under low oxygen conditions, distorting red blood cells into a
sickle shape1.
• Hemoglobin C (HbC):
• Chemical Structure: In HbC, glutamic acid at position 6 of the β-chain is replaced by lysine due to a point
mutation. This also alters hemoglobin's solubility and leads to mild chronic hemolytic anemia1.
 • Hemoglobin D-Punjab:
• Chemical Structure: Caused by a different point mutation in the β-chain, resulting in a variant hemoglobin with
altered properties1.
• Hemoglobin H (HbH):
• Chemical Structure: A tetramer of β-globin chains (β₄), typically seen in certain forms of α-thalassemia1.
• Other Forms:
• Methemoglobin: Iron in the heme group is oxidized to Fe³⁺, which cannot bind oxygen.
• Carboxyhemoglobin: Hemoglobin binds carbon monoxide instead of oxygen.
• Sulfhemoglobin: Hemoglobin irreversibly binds sulfur, rendering it incapable of oxygen transport1.
Sickle Cell Anemia
Molecular Basis:
Sickle cell anemia is caused by the production of hemoglobin S (HbS). The mutation substitutes valine for glutamic acid at
position 6 of the β-globin chain. This single amino acid change creates a hydrophobic region, promoting abnormal
polymerization of deoxygenated HbS and leading to the formation of rigid, sickle-shaped red blood cells1.
Consequences:
• Sickle-shaped cells are less flexible and can block small blood vessels, causing pain, organ damage, and increased risk of
infection.
• The abnormal cells are more prone to hemolysis, resulting in chronic anemia.
Clinical Features:
• Painful vaso-occlusive crises
• Hemolytic anemia
 • Increased risk of stroke, infections, and organ failure
Inheritance:
Sickle cell anemia is inherited in an autosomal recessive manner. Individuals with one mutated gene (heterozygotes) have
sickle cell trait, which is usually asymptomatic.
Thalassemia
Molecular Basis:
Thalassemias are a group of inherited disorders characterized by reduced or absent synthesis of one or more globin chains in
hemoglobin. The two main types are:
• α-Thalassemia:
• Caused by deletions or mutations in the genes encoding α-globin chains.
• Decreased α-chain production leads to an excess of β-chains, which can form unstable tetramers (HbH, β₄)1.
• β-Thalassemia:
• Caused by mutations in the β-globin gene, resulting in reduced (β⁺) or absent (β⁰) β-chain synthesis.
• Leads to an excess of α-chains, which precipitate and damage red blood cell precursors, causing ineffective
erythropoiesis and hemolytic anemia.
Consequences:
• Chronic anemia
• Bone marrow expansion and skeletal deformities
• Splenomegaly
• Growth retardation
Clinical Features:
 • Severity depends on the number and type of affected genes (minor, intermedia, major/Cooley's anemia).
• Severe forms require regular blood transfusions and iron chelation therapy.

16. Flavoproteins, chemical structure, representatives, biological role.

Ans. Chemical Structure
Flavoproteins are a class of complex (conjugated) proteins that contain a flavin nucleotide as their prosthetic group. The flavin
nucleotides can be either:
• Flavin mononucleotide (FMN)
• Flavin adenine dinucleotide (FAD)
These flavin groups are derived from riboflavin (vitamin B₂). The flavin prosthetic group is tightly (often covalently) bound to
the protein part (apoprotein), forming the active holoprotein. The unique isoalloxazine ring system of the flavin allows it to
participate in reversible redox (oxidation-reduction) reactions, accepting and donating electrons and protons14.
Representatives
Key examples of flavoproteins include:
• Succinate dehydrogenase (Complex II of the mitochondrial electron transport chain): Contains FAD and participates in
both the citric acid cycle and the respiratory chain.
• NADH dehydrogenase (Complex I): Contains FMN as a prosthetic group.
• Acyl-CoA dehydrogenase: Involved in fatty acid β-oxidation, contains FAD.
• Xanthine oxidase: Contains FAD, catalyzes the oxidation of xanthine to uric acid.
• Glucose oxidase: Contains FAD, used in glucose metabolism and diagnostic glucose assays.
 • L-amino acid oxidase: Contains FMN or FAD, catalyzes the oxidative deamination of amino acids.
• Cytochrome P450 reductase: Contains both FAD and FMN, essential for electron transfer to cytochrome P450
enzymes14.
Biological Role
Flavoproteins play crucial roles in a wide array of biological processes, primarily due to their ability to participate in redox
reactions:
• Biological Oxidation and Energy Production:
Flavoproteins are central to cellular respiration, especially in the mitochondrial electron transport chain, where they
facilitate the transfer of electrons from metabolic substrates to the respiratory chain, ultimately leading to ATP
synthesis346.
• Metabolism of Carbohydrates, Lipids, and Amino Acids:
Enzymes such as succinate dehydrogenase (citric acid cycle), acyl-CoA dehydrogenase (fatty acid oxidation), and various
amino acid oxidases depend on flavoproteins for their catalytic activity4.
• Detoxification and Drug Metabolism:
Flavoproteins like cytochrome P450 reductase are essential for the function of cytochrome P450 enzymes, which
catalyze the metabolism and detoxification of drugs and xenobiotics in the liver3.
• Antioxidant Defense and Removal of Free Radicals:
Flavoproteins such as glutathione reductase and NADPH oxidase are involved in the removal of reactive oxygen species,
contributing to cellular antioxidant defenses3.
• DNA Repair and Light Sensing:
Certain flavoproteins participate in DNA repair mechanisms and in photoreception (e.g., photolyases in DNA repair,
cryptochromes in circadian rhythm regulation)1.
17. Glycoproteins, chemical structure, representatives, biological role.
Ans. Chemical Structure
Glycoproteins are complex (conjugated) proteins in which the prosthetic group is a carbohydrate. The carbohydrate
component may consist of monosaccharides, oligosaccharides, or more complex sugar units such as hexoses (D-galactose, D-
mannose), deoxyhexoses (L-fucose, L-rhamnose), pentoses (D-xylose), amino sugars (N-acetyl-D-galactosamine, N-acetyl-D-
glucosamine, neuraminic acid), and uronic acids (D-glucuronic acid)1.
The carbohydrate moiety is covalently attached to the polypeptide chain via:
• O-linked glycosylation: Glycosidic bond to the hydroxyl group of serine (Ser) or threonine (Thr) residues.
• N-linked glycosylation: N-glycosyl bond to the amide nitrogen of asparagine (Asn) residues1.
Glycoproteins are found on the outer face of the plasma membrane, in the extracellular matrix, in the blood, and within
specific organelles such as the Golgi complex, secretory granules, and lysosomes1.
Representatives
Key examples of glycoproteins include1:
• Structural molecules: Collagens (in connective tissue)
• Lubricant and protective agents: Mucins (in mucus secretions)
• Transport molecules: Transferrin (iron transport), ceruloplasmin (copper transport)
• Immunologic molecules: Immunoglobulins (antibodies), histocompatibility antigens (MHC proteins)
• Hormones: Human chorionic gonadotropin (hCG), thyroid-stimulating hormone (TSH)
• Enzymes: Alkaline phosphatase, patatin
• Cell recognition and adhesion molecules: Lectins, selectins, and proteins involved in sperm-oocyte, virus-cell,
bacterium-cell, and hormone-cell interactions
 • Antifreeze proteins: Certain plasma proteins of cold-water fish
• Receptors: Proteins involved in hormone and drug action
• Protein folding regulators: Calnexin, calreticulin
• Developmental regulators: Notch and its analogs
• Hemostasis and thrombosis: Glycoproteins on platelet surfaces
Glycoproteins can be further divided by amino acid composition:
• Type I glycoproteins: Usual set of amino acids, carbohydrate content 3–4% (e.g., immunoglobulins, transferrin,
ceruloplasmin, thyroglobulin)
• Type II glycoproteins: High in serine, threonine, and proline, with 60–85% carbohydrate (e.g., glycoproteins of
submandibular gland secretion, group-specific antigens)1.
Biological Role
Glycoproteins perform a wide array of essential biological functions1:
• Structural: Provide strength and support in tissues (e.g., collagens in connective tissue)
• Protection and lubrication: Form protective barriers and lubricate surfaces (e.g., mucins in mucus)
• Transport: Bind and carry ions or molecules in the blood (e.g., transferrin for iron, ceruloplasmin for copper)
• Immunity: Serve as antibodies and as antigens for immune recognition (e.g., immunoglobulins, MHC proteins)
• Hormonal regulation: Act as hormones or hormone receptors (e.g., TSH, hCG)
• Enzymatic activity: Many enzymes are glycoproteins, contributing to metabolic processes
• Cell recognition and adhesion: Mediate cell-cell interactions, recognition, and signaling (e.g., lectins, selectins)
• Antifreeze activity: Prevent freezing in the blood of certain fish
 • Protein folding and quality control: Assist in the proper folding of newly synthesized proteins (e.g., calnexin, calreticulin)
• Regulation of development: Involved in developmental processes (e.g., Notch signaling)
• Hemostasis and thrombosis: Involved in blood clotting and platelet function
Glycoproteins are thus indispensable for structural integrity, communication, immunity, metabolism, and development in
multicellular organisms1.

18. Lipoproteins, structural and transport. Classification of transport forms.

Ans. Structural and Transport Lipoproteins
Lipoproteins are complex conjugated proteins whose prosthetic group is represented by lipids. They are essential for the
transport of hydrophobic lipids (like triglycerides, cholesterol, and phospholipids) in the aqueous environment of blood
plasma.
Structure
• Core: Hydrophobic, consisting mainly of cholesterol esters and triglycerides.
• Surface: Hydrophilic, composed of phospholipids, free cholesterol, and specific proteins called apolipoproteins.
• This amphipathic structure allows lipoproteins to solubilize and transport lipids through the bloodstream1.
Types
• Structural Lipoproteins: These are integral parts of biological membranes (transmembrane lipoproteins) and contribute
to membrane structure and function.
• Transport Lipoproteins: These are plasma lipoprotein particles responsible for the transport of lipids between organs
and tissues1.
Classification of Transport Forms of Lipoproteins
Plasma lipoproteins are classified based on their size, density, lipid composition, and apolipoprotein content. The main classes
are:
1. Chylomicrons
• Diameter: 75–1200 nm (largest)
• Function: Carry dietary triglycerides and cholesterol from the intestines to the liver, skeletal muscle, and adipose tissue.
• Apolipoproteins: ApoB-48, ApoC, ApoE1.
2. Very-Low-Density Lipoproteins (VLDL)
• Diameter: 30–80 nm
• Function: Transport newly synthesized triglycerides from the liver to adipose tissue.
• Apolipoproteins: ApoB-100, ApoC, ApoE1.
3. Intermediate-Density Lipoproteins (IDL)
• Diameter: 25–50 nm
• Function: Transitional form between VLDL and LDL; not usually detectable in fasting blood.
• Apolipoproteins: ApoB-100, ApoE1.
4. Low-Density Lipoproteins (LDL)
• Diameter: 18–28 nm
• Function: Deliver cholesterol to peripheral tissues; often referred to as "bad cholesterol" due to its association with
atherosclerosis.
• Apolipoproteins: ApoB-1001.
5. High-Density Lipoproteins (HDL)
 • Diameter: 5–15 nm (smallest)
• Function: Collect cholesterol from tissues and transport it back to the liver for excretion ("reverse cholesterol
transport"); known as "good cholesterol."
• Apolipoproteins: ApoA-I, ApoA-II, ApoC, ApoE1.

19. Metalloproteins, chemical structure, representatives, biological role.

Ans. Chemical Structure
Metalloproteins are complex (conjugated) proteins that contain one or more metal ions as their prosthetic group. The metal
ion is tightly bound to the protein, often coordinated by specific amino acid side chains (such as histidine, cysteine, aspartate,
or glutamate) or by non-protein ligands. The protein part is called the apoprotein; together with the metal ion, it forms the
functional holoprotein1.
The metal ion in metalloproteins is essential for the protein’s structure and/or biological activity. These metal ions can
participate directly in catalysis, stabilize protein structure, or facilitate the binding and transport of molecules.
Representatives
Common examples of metalloproteins and their associated metal ions include:
• Iron (Fe):
• Ferritin: Iron storage protein in the liver and other tissues.
• Hemoglobin and Myoglobin: While primarily considered hemoproteins, they are also metalloproteins due to their
iron content.
• Iron-sulfur proteins: Involved in electron transport and redox reactions.
• Zinc (Zn):
 • Alcohol dehydrogenase: Catalyzes the oxidation of alcohols.
• Carbonic anhydrase: Catalyzes the reversible hydration of carbon dioxide.
• Carboxypeptidases: Digestive enzymes.
• Copper (Cu):
• Ceruloplasmin: Major copper-carrying protein in blood; involved in iron metabolism.
• Superoxide dismutase: Protects cells from oxidative damage by catalyzing the dismutation of superoxide radicals.
• Calcium (Ca):
• Calmodulin: Calcium-binding protein that regulates various cellular processes.
• Molybdenum (Mo):
• Dinitrogenase: Involved in nitrogen fixation in bacteria.
• Cobalt (Co):
• Vitamin B12-dependent enzymes: Such as methylmalonyl-CoA mutase.
• Nickel (Ni):
• Urease: Catalyzes the hydrolysis of urea.
Biological Role
Metalloproteins have a wide range of biological functions, including1:
• Transport:
• Ferritin stores and releases iron as needed.
• Ceruloplasmin transports copper and assists in iron metabolism.
 • Catalysis (Metalloenzymes):
• Carbonic anhydrase (Zn) speeds up CO₂ hydration.
• Alcohol dehydrogenase (Zn) is key in alcohol metabolism.
• Superoxide dismutase (Cu, Zn) protects against oxidative stress.
• Dinitrogenase (Mo) is crucial for nitrogen fixation.
• Structural:
• Metal ions can stabilize protein structures, as in calmodulin (Ca).
• Reserve:
• Ferritin acts as a reserve for iron, releasing it when needed.
• Regulation:
• Calmodulin (Ca) regulates many cellular enzymes and processes by binding calcium ions.
• Electron Transport and Redox Reactions:
• Iron-sulfur proteins and cytochromes (Fe) are essential in the electron transport chain and cellular respiration.

20. Phosphoproteins, chemical structure, representatives, biological role.

Ans. Chemical Structure
Phosphoproteins are a class of complex (conjugated) proteins that contain phosphoric acid (phosphate groups) as their
prosthetic group. The phosphate group is covalently attached to the hydroxyl groups of specific amino acids-
primarily serine, threonine, and tyrosine-within the polypeptide chain. This attachment forms phosphoester bonds (O-
phosphorylation).
 • The protein part is called the apoprotein.
• The complete molecule, with phosphate groups, is the holoprotein.
• Phosphorylation is a reversible post-translational modification, often catalyzed by protein kinases (which add
phosphate) and removed by phosphatases (which remove phosphate)26.
Representatives of Phosphoproteins
Key examples of phosphoproteins include:
• Casein: The main protein in milk; rich in phosphoserine residues. It binds calcium and phosphate, aiding in their
transport and absorption2.
• Vitellin: Found in egg yolk; serves as a nutrient source for developing embryos and is rich in phosphate.
• Phosvitin: A highly phosphorylated protein in egg yolk, important for metal ion binding.
• Ovalbumin: The main protein in egg white, which also contains phosphate groups.
• Enzymes: Many enzymes are regulated by phosphorylation, such as glycogen phosphorylase and several metabolic
enzymes6.
Biological Role
Phosphoproteins serve several crucial functions in the body:
• Nutritional Role:
• Casein provides a source of essential amino acids, calcium, and phosphate for infants and young animals. The
phosphate groups allow casein to bind calcium and keep it soluble, facilitating its absorption from the digestive
tract2.
• Vitellin and phosvitin supply phosphate, amino acids, and metals for the developing embryo.
• Structural Role:
 • Phosphorylation can alter the structural conformation of proteins, affecting their stability, solubility, and
interactions with other molecules.
• Regulatory Role:
• Protein phosphorylation is a central mechanism for regulating enzyme activity, signal transduction, and cellular
processes. The addition or removal of phosphate groups can activate or deactivate enzymes and other proteins,
thus controlling metabolic pathways, cell division, growth, and differentiation26.
• This reversible modification is essential for cellular responses to hormones, growth factors, and other signals.
• Catalytic Role:
• Many enzymes require phosphorylation for activation or regulation. For example, phosphorylation of metabolic
21. Polymerase chain reaction. The principle of the method. Application in medicine.
Ans. Principle of the Method
The polymerase chain reaction (PCR) is a molecular biology technique used to amplify specific DNA sequences, making
millions to billions of copies of a particular DNA segment from a small initial sample. The method relies on the ability of DNA
polymerase to synthesize new strands of DNA complementary to the target sequence.
Key Steps in PCR:
1. Denaturation: The double-stranded DNA is heated (typically to 94–98°C) to separate it into two single strands.
2. Annealing: The reaction is cooled (usually to 50–65°C) to allow short DNA primers-synthetic oligonucleotides designed
to flank the target region-to bind (anneal) to their complementary sequences on the single-stranded DNA.
3. Extension (Elongation): The temperature is raised to the optimum for DNA polymerase (commonly 72°C), which
synthesizes new DNA strands by extending from the primers, using the original strands as templates.
These three steps constitute one cycle, and the process is repeated for 25–40 cycles, resulting in exponential amplification of
the target DNA segment.
Essential Components:
• Template DNA (containing the target sequence)
• Two primers (forward and reverse)
• DNA polymerase (commonly Taq polymerase, which is heat-stable)
• Deoxynucleotide triphosphates (dNTPs: A, T, G, C)
• Buffer solution with magnesium ions
Applications in Medicine
PCR has revolutionized medical diagnostics and research due to its sensitivity, specificity, and speed. Major applications in
medicine include:
• Infectious Disease Diagnosis:
PCR can detect the genetic material of pathogens (viruses, bacteria, fungi) directly from clinical samples, enabling rapid
diagnosis of diseases such as HIV, hepatitis, tuberculosis, COVID-19, and many others-even when only a few organisms
are present.
• Genetic and Hereditary Disease Testing:
PCR is used to identify mutations, deletions, or insertions in genes associated with inherited disorders (e.g., cystic
fibrosis, sickle cell anemia, thalassemia).
• Oncology:
Detection of cancer-associated gene mutations, minimal residual disease, and monitoring of treatment response (e.g.,
BCR-ABL fusion gene in chronic myeloid leukemia).
• Forensic Medicine:
PCR enables DNA fingerprinting for individual identification in forensic investigations, paternity testing, and disaster
victim identification.
 • Prenatal and Neonatal Screening:
Early detection of genetic diseases in fetuses and newborns using PCR-based assays.
• Transplant Medicine:
Detection of donor and recipient compatibility, and monitoring for infections or graft rejection.
• Pharmacogenomics:
Assessing genetic variations that affect drug metabolism, guiding personalized medicine.

22. Enzymes. Classification, nomenclature, code.

Ans. Definition and General Features
Enzymes are biocatalysts-primarily proteins (with rare exceptions such as catalytic RNAs)-that accelerate chemical reactions in
the body without being consumed in the process. They act by lowering the activation energy required for a reaction, thereby
increasing its rate. Enzymes are highly specific for their substrates and reactions, operate under mild physiological conditions,
and are subject to complex regulation1.
Classification of Enzymes
The international system for enzyme classification is established by the Enzyme Commission (EC) and groups enzymes into
seven main classes, based on the type of reaction they catalyze1:
1. Oxidoreductases (EC 1):
• Catalyze oxidation-reduction (redox) reactions (transfer of electrons or hydrogen atoms).
• Example: Lactate dehydrogenase.
2. Transferases (EC 2):
• Transfer functional groups (e.g., methyl, phosphate, amino) from one molecule to another.
 • Example: Alanine aminotransferase.
3. Hydrolases (EC 3):
• Catalyze hydrolysis reactions (cleavage of bonds by addition of water).
• Example: Trypsin, lipase.
4. Lyases (EC 4):
• Catalyze the addition or removal of groups to form double bonds, or the breaking of various bonds by means
other than hydrolysis and oxidation.
• Example: Fumarase.
5. Isomerases (EC 5):
• Catalyze isomerization changes within a single molecule (rearrangement of atoms).
• Example: Phosphoglucose isomerase.
6. Ligases (EC 6):
• Join two molecules together with covalent bonds, often coupled with the hydrolysis of ATP.
• Example: DNA ligase.
7. Translocases (EC 7):
• Catalyze the movement of ions or molecules across membranes or their separation within membranes.
• Example: ATP synthase1.
Nomenclature of Enzymes
Enzyme names typically end with the suffix -ase and are often descriptive of the substrate acted upon and/or the type of
reaction catalyzed. There are two naming systems1:
 • Trivial Names: Common, traditional names (e.g., trypsin, pepsin).
• Systematic Names: Specify the substrate(s) and the type of reaction (e.g., lactate dehydrogenase for the enzyme that
removes hydrogen from lactate).
For example, lactate dehydrogenase catalyzes the oxidation of lactate to pyruvate.
Enzyme Commission (EC) Code
Each enzyme is assigned a unique Enzyme Commission (EC) number, which consists of four numbers separated by periods:
• First number: Main class (type of reaction)
• Second number: Subclass (type of group or bond involved)
• Third number: Sub-subclass (more specific details)
• Fourth number: Serial number of the enzyme in this sub-subclass
Example:
ATP:glucose phosphotransferase (hexokinase)
• EC 2.7.1.1
• 2 = Transferase
• 7 = Transfers a phosphate group
• 1 = Phosphotransferase with a hydroxyl group as acceptor
• 1 = Glucose as the acceptor

23. Structural organization of enzymes. Active, allosteric site. Functional groups of enzymes.
Ans. Structural Organization of Enzymes
Enzymes are primarily proteins (with rare exceptions like catalytic RNAs) that possess a complex, hierarchical structure
essential for their catalytic function. Their organization includes:
• Apoenzyme: The protein portion of the enzyme, responsible for the enzyme's specificity and catalytic activity.
• Holoenzyme: The complete, catalytically active enzyme, which consists of the apoenzyme plus any required cofactors or
coenzymes (e.g., metal ions, organic molecules)1.
• Prosthetic Group: A tightly or covalently bound non-protein component (e.g., heme, flavin) if present.
The enzyme's structure creates a specific three-dimensional conformation, which forms the basis for the enzyme's functional
sites.
Active Site
The active site is a specialized region on the enzyme where substrate binding and catalysis occur. Key features include:
• Location: The active site is typically a pocket or groove formed by the enzyme's tertiary structure.
• Composition: It is lined with specific amino acid residues whose side chains participate in substrate binding and the
catalytic reaction.
• Function: The active site binds the substrate(s) with high specificity and orients them for the chemical reaction. It lowers
the activation energy, allowing the reaction to proceed more rapidly.
• Binding and Catalytic Sites: The active site may be subdivided into a binding site (which holds the substrate) and a
catalytic site (where the chemical transformation occurs)1.
Models of Substrate Binding:
• Lock-and-Key Model: The enzyme and substrate have complementary shapes that fit exactly.
• Induced Fit Model: The enzyme structure is flexible and molds itself around the substrate upon binding, enhancing
specificity and catalytic efficiency1.
Allosteric Site
The allosteric site is a distinct region on the enzyme, spatially separated from the active site. Its features include:
• Function: Binds regulatory molecules called allosteric effectors (activators or inhibitors).
• Mechanism: Binding of an effector at the allosteric site induces a conformational change in the enzyme, which alters the
configuration of the active site and modulates enzyme activity (either increasing or decreasing reaction velocity).
• Role in Regulation: Allosteric regulation is a key mechanism for controlling enzyme activity in metabolic pathways,
allowing feedback inhibition or activation in response to cellular needs1.
Functional Groups of Enzymes
The catalytic activity of enzymes depends on the presence of specific functional groups in the amino acid side chains at the
active site. These groups can:
• Participate in Acid-Base Catalysis: Amino acids such as glutamic acid (Glu), aspartic acid (Asp), lysine (Lys), arginine
(Arg), histidine (His), cysteine (Cys), serine (Ser), and tyrosine (Tyr) can act as proton donors or acceptors during the
reaction.
• Form Covalent Intermediates: Some side chains (e.g., Ser, Cys, His) can form transient covalent bonds with the
substrate.
• Stabilize Transition States: Through hydrogen bonding, ionic interactions, and hydrophobic effects, the active site
residues stabilize the transition state of the substrate, facilitating the reaction1.
Examples of Functional Groups:
• Carboxyl (-COOH / -COO⁻): Glu, Asp
• Amino (-NH₂ / -NH₃⁺): Lys, Arg
• Imidazole (from His): Can act as both acid and base
 • Thiol (-SH): Cys
• Hydroxyl (-OH): Ser, Tyr

24. Enzyme properties. Dependence of the rate of enzymatic reactions on pH, temperature. Enzyme specificity.
Ans. Enzyme Properties
Enzymes are biological catalysts, almost always proteins, that accelerate chemical reactions in living systems without being
consumed or permanently altered in the process. Their key properties include:
• Catalytic efficiency: Enzymes increase reaction rates by factors of 10⁶–10¹² compared to uncatalyzed reactions.
• Specificity: Enzymes are highly specific for their substrates and the type of reaction they catalyze.
• Mild conditions: They function effectively under physiological conditions-neutral pH, moderate temperatures, and
aqueous environments.
• Regulation: Enzyme activity can be finely regulated by various mechanisms, including allosteric control, covalent
modification, and changes in enzyme quantity.
• Not consumed: Enzymes are not used up during the reaction and can catalyze multiple cycles1.
Dependence of Enzymatic Reaction Rate on pH
• Optimal pH: Each enzyme has an optimal pH at which its activity is maximal. This is related to the ionization state of
amino acid residues at the active site and the substrate.
• Activity loss at extreme pH: Deviations from the optimal pH can lead to decreased activity or denaturation due to
altered charges on amino acids, disrupting enzyme structure or substrate binding.
• Examples:
• Pepsin (stomach): Optimum pH ~1.5–2
 • Trypsin (intestine): Optimum pH ~7.5–8.5
• Denaturation: Extreme pH values can irreversibly denature enzymes, destroying their catalytic activity14.
Dependence of Enzymatic Reaction Rate on Temperature
• Optimal temperature: Each enzyme has an optimal temperature (often around 37°C in humans) where activity is
highest.
• Temperature increase: Raising temperature generally increases reaction rate due to higher kinetic energy-up to the
optimum.
• Denaturation at high temperatures: Above the optimal temperature, enzyme structure is disrupted (denatured), leading
to rapid loss of activity.
• Low temperatures: Lowering temperature slows reaction rates but usually does not denature the enzyme, so activity
can be restored by returning to optimal conditions14.
Enzyme Specificity
Enzymes display several types of specificity:
• Bond specificity: Some enzymes act only on specific types of chemical bonds (e.g., peptidases on peptide bonds).
• Group specificity: Some enzymes act on substrates with particular functional groups (e.g., alcohol dehydrogenase acts
on alcohols).
• Substrate specificity: Many enzymes act only on a single substrate (e.g., urease on urea).
• Optical (stereospecificity): Enzymes often distinguish between optical isomers (L- or D- forms).
• Geometric specificity: Some enzymes recognize specific geometric configurations of substrates.
Models of specificity:
• Lock-and-key model: The enzyme’s active site is a rigid structure complementary to the substrate.
 • Induced fit model: The enzyme’s active site is flexible and molds itself around the substrate upon binding, increasing
specificity and catalytic efficiency1.

25. The mechanism of action of enzymes. Enzymatic catalysis theories.

Ans. Mechanism of Action of Enzymes
Enzymes are biological catalysts that accelerate chemical reactions by lowering the activation energy required for the reaction
to proceed. They do this without being consumed or permanently altered in the process. The general mechanism involves
several key steps:
1. Substrate Binding:
The substrate (reactant) binds to a specific region of the enzyme called the active site. This site is a three-dimensional
pocket formed by the enzyme’s tertiary structure and lined with amino acid residues that interact with the substrate
through non-covalent forces (hydrogen bonds, ionic bonds, hydrophobic interactions, van der Waals forces)1.
2. Formation of the Enzyme-Substrate Complex (ES):
The binding of the substrate to the enzyme forms a transient enzyme-substrate complex (ES), which positions the
substrate(s) in an optimal orientation for the reaction to occur1.
3. Catalysis (Transition State Stabilization):
The enzyme stabilizes the transition state of the reaction, effectively lowering the activation energy. This can involve:
• Bringing substrates into close proximity (proximity effect)
• Orienting substrates correctly (orientation effect)
• Providing acidic or basic groups for proton transfer (acid-base catalysis)
• Forming transient covalent intermediates (covalent catalysis)
• Distorting the substrate to facilitate bond breaking/forming (strain effect)1.
 4. Product Formation and Release:
After the reaction, the enzyme releases the product(s) and returns to its original state, ready to catalyze another
reaction cycle1.
The overall reaction can be summarized as:
E+S⇌ES→EP⇌E+PE+S⇌ES→EP⇌E+P
where EE is enzyme, SS is substrate, ESES is enzyme-substrate complex, EPEP is enzyme-product complex, and PP is product.
Theories of Enzymatic Catalysis
1. Lock-and-Key Model (Emil Fischer, 1894)
• Proposes that the enzyme’s active site has a specific, rigid shape that exactly fits the substrate, much like a key fits into a
lock.
• This model explains the high specificity of enzymes but does not account for the flexibility of enzyme structures or the
stabilization of the transition state1.
2. Induced Fit Model (Daniel Koshland, 1958)
• Suggests that the active site is flexible and can change its shape to fit the substrate more closely upon binding.
• The enzyme undergoes a conformational change, molding itself around the substrate, which enhances binding and
catalytic efficiency.
• This model better explains enzyme specificity, transition state stabilization, and the dynamic nature of enzyme-substrate
interactions1.

26. Enzyme activators and inhibitors. Inhibition types. Enzyme activation and inactivation mechanisms.
Ans. Enzyme Activators and Inhibitors
Enzyme activators are molecules that increase the activity of an enzyme. They often work by binding to the enzyme and
inducing a conformational change that enhances substrate binding or catalytic efficiency. Common activators include certain
metal ions (e.g., Mg²⁺, Ca²⁺, Zn²⁺) and small organic molecules. For example, ADP acts as an allosteric activator for
phosphofructokinase-1, a key regulatory enzyme in glycolysis1.
Enzyme inhibitors are substances that decrease or completely block enzyme activity. Inhibitors can be endogenous (produced
within the body) or exogenous (drugs, toxins). Inhibition can be reversible or irreversible, depending on the nature of the
interaction with the enzyme16.
Types of Enzyme Inhibition
1. Competitive Inhibition:
A competitive inhibitor resembles the substrate and competes for binding at the enzyme’s active site. This type of inhibition
can be overcome by increasing substrate concentration. The inhibitor does not affect the maximum velocity (Vmax) but
increases the apparent Km (Michaelis constant), meaning a higher substrate concentration is needed to reach half-maximal
velocity.
2. Noncompetitive Inhibition:
A noncompetitive inhibitor binds to a site other than the active site (allosteric site), causing a conformational change that
reduces enzyme activity. This inhibition cannot be overcome by increasing substrate concentration. Vmax decreases, but Km
remains unchanged.
3. Uncompetitive Inhibition:
The inhibitor binds only to the enzyme-substrate complex, locking the substrate in the enzyme and preventing the reaction.
Both Vmax and Km decrease.
4. Irreversible Inhibition:
The inhibitor forms a covalent bond with the enzyme, permanently inactivating it. Examples include certain toxins and drugs
(e.g., organophosphates inhibiting acetylcholinesterase)16.
5. Allosteric Inhibition:
Allosteric inhibitors bind to a regulatory (allosteric) site, distinct from the active site, causing a conformational change that
reduces enzyme activity. This is a key mechanism for feedback regulation in metabolic pathways1.
Enzyme Activation and Inactivation Mechanisms
1. Allosteric Regulation:
Enzyme activity is modulated by the binding of activators or inhibitors at allosteric sites. Allosteric activators increase enzyme
activity (e.g., AMP activating phosphofructokinase-1), while allosteric inhibitors decrease it (e.g., ATP or citrate inhibiting
phosphofructokinase-1)12.
2. Covalent Modification:
Enzymes can be activated or inactivated by the addition or removal of specific chemical groups, most commonly phosphate
groups (phosphorylation/dephosphorylation). Protein kinases add phosphate groups, often activating or inactivating enzymes,
while phosphatases remove them12.
3. Zymogen (Proenzyme) Activation:
Some enzymes are synthesized and secreted as inactive precursors (zymogens or proenzymes) and are activated by specific
cleavage of peptide bonds. For example, pepsinogen is activated to pepsin in the stomach by hydrochloric acid; trypsinogen is
activated to trypsin by enterokinase in the intestine14.
4. Feedback Inhibition:
The end product of a metabolic pathway inhibits an enzyme involved earlier in the pathway, preventing overaccumulation of
the product. This is a rapid and reversible mechanism of metabolic regulation12.
5. Protein-Protein Interactions:
Some enzymes are regulated through interactions with other proteins. For example, regulatory and catalytic subunits may
associate or dissociate to modulate enzyme activity (e.g., protein kinase A)1.

27. Enzyme specificity. Views.

Ans. Enzyme specificity refers to the remarkable ability of enzymes to selectively recognize, bind, and catalyze the
transformation of specific substrates or types of chemical bonds. This property distinguishes enzymes from inorganic catalysts
and underlies the precision of biochemical processes in living organisms1.
Types of Enzyme Specificity
Enzymes exhibit several distinct types of specificity:
• Bond Specificity:
The enzyme acts only on a specific type of chemical bond, regardless of the rest of the molecular structure. For example,
peptidases specifically hydrolyze peptide bonds between amino acids, regardless of which amino acids are involved1.
• Group Specificity:
The enzyme acts on molecules that have a particular functional group. For instance, alcohol dehydrogenase acts on
alcohols, oxidizing their hydroxyl groups, but does not act on other types of compounds1.
• Substrate Specificity:
The enzyme acts only on a particular substrate. Urease, for example, catalyzes the hydrolysis of urea but not of other
amides1.
• Optical (Stereospecificity):
Many enzymes can distinguish between optical isomers (enantiomers) and will act only on one form. For example, most
enzymes in the human body act only on L-amino acids, not D-amino acids1.
• Geometric Specificity:
Some enzymes recognize and act only on substrates with a specific geometric configuration, such as cis or trans isomers.
For example, fumarase acts only on the trans isomer fumarate and not on the cis isomer maleate1.
• Co-factor Specificity:
Some enzymes require a specific cofactor (such as a particular metal ion or coenzyme) to function and will not work with
substitutes1.
Views and Theories Explaining Enzyme Specificity
1. Lock-and-Key Model (Emil Fischer, 1894):
This classic model proposes that the enzyme’s active site has a specific, rigid shape complementary to the substrate, much like
a key fits into a lock. Only substrates with the correct shape can bind and be converted to product. This model explains the
high specificity of enzymes but does not account for enzyme flexibility1.
2. Induced Fit Model (Daniel Koshland, 1958):
This more modern view suggests that the enzyme’s active site is flexible and molds itself around the substrate upon binding.
The binding of the substrate induces a conformational change in the enzyme, optimizing the fit and facilitating catalysis. This
model explains both specificity and the ability of enzymes to stabilize the transition state of the reaction1.

28. Ways of regulation of enzyme activity in the cell: change in the number of enzyme molecules, availability of substrate
molecules and coenzymes, allosteric regulation. Regulation of the catalytic activity of enzymes by protein-protein interaction,
by phosphorylation, dephosphorylation, limited proteolysis.
Ans. Regulation of Enzyme Activity in the Cell
Enzyme activity in cells is tightly regulated to ensure metabolic pathways function efficiently and adapt to changing
physiological needs. Regulation can occur by altering the amount of enzyme present, the availability of substrates and
cofactors, and by directly modifying the enzyme's catalytic activity through several mechanisms.
1. Change in the Number of Enzyme Molecules
• Synthesis and Degradation:
The quantity of enzyme in the cell depends on the rates of its synthesis (gene expression) and degradation.
• Induction increases enzyme synthesis in response to specific signals (e.g., hormones, substrate presence).
• Repression decreases enzyme synthesis, often via feedback from pathway end products.
 • This regulation is relatively slow, taking hours to days to manifest, and is crucial for long-term metabolic
adaptation2.
2. Availability of Substrate Molecules and Coenzymes
• Substrate Concentration:
The rate of an enzyme-catalyzed reaction depends on substrate availability. Increased substrate concentration generally
increases reaction rate until the enzyme becomes saturated.
• Coenzymes and Metal Ions:
Many enzymes require coenzymes (organic molecules) or metal ions as cofactors for activity. Their presence or absence
can activate or inhibit enzyme function. For example, Mg²⁺ is required for kinases, and NAD⁺/NADH ratios regulate many
dehydrogenases2.
3. Allosteric Regulation
• Allosteric Sites:
Many enzymes have regulatory sites (allosteric sites) distinct from the active site. Binding of allosteric effectors
(activators or inhibitors) induces conformational changes that alter enzyme activity.
• Positive Effectors (Activators):
Enhance enzyme activity. For example, AMP is an allosteric activator of phosphofructokinase-1 in glycolysis.
• Negative Effectors (Inhibitors):
Decrease enzyme activity. ATP and citrate inhibit phosphofructokinase-1, signaling sufficient energy and reducing
glycolytic flux.
• Allosteric regulation allows rapid, reversible control of enzyme activity in response to cellular signals and metabolic
needs2.
4. Protein-Protein Interactions
• Some enzymes are regulated by association or dissociation with other protein subunits.
 • Example: Protein kinase A is inactive as a holoenzyme (regulatory and catalytic subunits together). Activation occurs
when cAMP binds regulatory subunits, releasing active catalytic subunits.
• This mechanism allows for rapid, signal-dependent modulation of enzyme activity2.
5. Phosphorylation and Dephosphorylation (Covalent Modification)
• Phosphorylation:
Addition of phosphate groups (usually to serine, threonine, or tyrosine residues) by protein kinases can activate or
inactivate enzymes.
• Dephosphorylation:
Removal of phosphate groups by phosphatases reverses the modification.
• This regulation is rapid and reversible, often under hormonal control (e.g., insulin, glucagon, epinephrine).
Example: Phosphorylation inactivates pyruvate kinase in glycolysis, inhibiting the pathway when energy is abundant21.
6. Limited Proteolysis (Zymogen Activation)
• Some enzymes are synthesized as inactive precursors (zymogens or proenzymes) and activated by specific proteolytic
cleavage.
• This is common for digestive enzymes (e.g., trypsinogen to trypsin) and blood coagulation factors.
• Activation by proteolysis is irreversible and ensures enzymes are active only when and where needed2.

29. Principles for the quantification of enzymes. Activity units.

Ans. Principles for the Quantification of Enzymes and Activity Units
Principles of Enzyme Quantification
Enzyme quantification is based on measuring the catalytic activity of an enzyme, rather than the amount of protein present.
This is because the biological function of an enzyme is directly related to its ability to catalyze a specific chemical reaction. The
standard approach is to measure either the rate of substrate consumption or the rate of product formation over time under
defined conditions.
Key principles include:
• Enzyme assays are laboratory methods designed to measure enzyme activity. They can be direct (measuring product
formation or substrate disappearance in the reaction mixture) or indirect (measuring a secondary reaction that depends
on the enzyme's activity).
• The reaction is performed under optimal and standardized conditions (temperature, pH, substrate concentration, ionic
strength) to ensure reproducibility and comparability.
• The initial reaction rate (when substrate concentration is much greater than enzyme concentration, and product
accumulation is minimal) is used to avoid complications from product inhibition or substrate depletion.
• The amount of enzyme is expressed in terms of its activity, not mass, since different enzyme preparations may have
different specific activities due to purity or presence of inhibitors.
Activity Units
Enzyme activity is expressed in standardized units that reflect the amount of substrate converted or product formed per unit
time under specified conditions.
• Enzyme Unit (U):
One unit (U) is defined as the amount of enzyme that catalyzes the conversion of 1 micromole (μmol) of substrate per
minute under specified assay conditions (e.g., temperature, pH, substrate concentration)1.
• Katal (kat):
The katal is the SI unit of enzyme activity. One katal is the amount of enzyme that converts 1 mole of substrate per
second under specified conditions.
 The relationship between units:
1 U=16.67 nkat1U=16.67nkat (nanokatal)1.
Factors to control in enzyme assays:
• Temperature
• pH
• Substrate concentration (ideally at saturation)
• Salt and buffer composition
• Presence of activators or inhibitors
Enzyme activity can be measured by various methods:
• Spectrophotometric: Monitoring changes in absorbance due to substrate or product.
• Gasometric: Measuring gas produced or consumed.
• Chromatographic: Separating and quantifying products.
• Polarimetric: Measuring changes in optical rotation.
Clinical and Research Relevance
• Enzyme activity measurement is fundamental in diagnostic enzymology (e.g., measuring ALT, AST, amylase, lipase in
serum to diagnose tissue damage or disease)1.
• Enzymes are also used as reagents in clinical chemistry for the quantification of metabolites (e.g., glucose oxidase for
glucose measurement).

30. Enzyme identification methods: chemical, рolarimetric, gasometric, chromatography, spectrophotometric

Ans. Enzyme Identification Methods: Chemical, Polarimetric, Gasometric, Chromatography, Spectrophotometric
Several methods are used to identify and quantify enzymes based on their catalytic activity or specific properties. Here is an
overview of the main approaches:
1. Chemical Methods
Chemical methods involve measuring the amount of substrate consumed or product formed during an enzyme-catalyzed
reaction using specific chemical reactions. For example, the product may react with a reagent to form a colored compound,
which can then be quantified. Classic examples include the use of the Biuret or Lowry methods for protein determination, or
colorimetric assays for enzyme activity (such as the use of dinitrosalicylic acid to detect reducing sugars in amylase assays)2.
2. Polarimetric Methods
Polarimetric methods are based on the ability of optically active substances (such as sugars or amino acids) to rotate the plane
of polarized light. Enzyme reactions that convert one optical isomer to another (e.g., glucose to fructose, or L- to D-amino
acids) can be monitored by measuring changes in optical rotation. The rate of change in optical rotation is directly proportional
to enzyme activity2.
3. Gasometric Methods
Gasometric methods measure the volume of gas produced or consumed during an enzyme-catalyzed reaction. For example,
urease catalyzes the breakdown of urea to produce ammonia and carbon dioxide; catalase breaks down hydrogen peroxide to
release oxygen. The amount of gas evolved or absorbed is measured and used to calculate enzyme activity2.
4. Chromatography
Chromatographic methods separate enzymes or their reaction products based on differences in physical or chemical properties
such as size, charge, or affinity. Types include:
• Ion-exchange chromatography: Separates proteins by charge.
• Size-exclusion (gel filtration) chromatography: Separates by molecular size.
 • Affinity chromatography: Separates based on specific binding to a ligand.
Chromatography is used both for enzyme purification and for identification by analyzing the presence and activity of enzymes
in different fractions2.
5. Spectrophotometric Methods
Spectrophotometric methods are among the most widely used for enzyme identification and quantification. They rely on
measuring the change in absorbance of light at a specific wavelength as a substrate is converted to product (or vice versa). This
can be due to the appearance or disappearance of a colored compound, or a change in absorbance in the ultraviolet or visible
range.
• Direct spectrophotometry: Measures the absorbance of the substrate or product directly.
• Coupled assays: Use an additional reaction to produce a measurable change in absorbance.
Enzyme activity is calculated from the rate of change in absorbance, using the Beer-Lambert law2.
31. Enzyme therapy.
Ans. Enzyme Therapy
Enzyme therapy refers to the medical use of enzymes as therapeutic agents to treat diseases caused by enzyme deficiencies,
metabolic disorders, or to achieve specific pharmacological effects. This approach can involve replacing missing or deficient
enzymes, supplementing digestive enzymes, or using enzymes to break down pathological substances in the body.
Principles and Types of Enzyme Therapy
1. Enzyme Replacement Therapy (ERT)
• This is the primary form of enzyme therapy, where a specific enzyme that is deficient or absent in the body is
replaced by administering it as a drug.
• ERT is especially important in treating inherited metabolic disorders (enzymopathies) where a genetic defect leads
to the absence or malfunction of a particular enzyme.
 • Example: Administration of pancreatic enzymes (e.g., Creon, Pancrease, Nutrizym, Pancrex) in pancreatic
insufficiency, or replacement of lysosomal enzymes in diseases like Gaucher's or Fabry's disease1.
2. Digestive Enzyme Therapy
• Used in gastrointestinal diseases associated with insufficient secretion of digestive juices, such as pepsin in
achylia, hypo- and anacid gastritis.
• Pancreatic enzyme preparations help in the digestion of proteins, fats, and carbohydrates in patients with
pancreatic insufficiency1.
3. Proteolytic Enzyme Therapy
• Proteolytic enzymes like trypsin and chymotrypsin are used to treat septic wounds by breaking down the proteins
of dead cells, aiding in wound cleaning and healing.
• They are also used to remove blood clots or viscous secretions in inflammatory respiratory diseases1.
4. Enzyme Therapy in Thrombolysis
• Enzymes such as fibrinolysin, streptokinase, streptodecase, and urokinase are used to dissolve blood clots in cases
of thrombosis and thromboembolism1.
5. Enzyme Therapy in Ophthalmology and Scar Treatment
• Ribonuclease and deoxyribonuclease are used as antiviral drugs in the treatment of adenoviral conjunctivitis and
herpetic keratitis.
• Hyaluronidase (lidase), which catalyzes the cleavage of hyaluronic acid, is used subcutaneously or intramuscularly
to promote the resorption of contractures and scars after burns or surgery1.
Clinical Applications and Examples
• Gastrointestinal Disorders: Supplementation with digestive enzymes (e.g., pepsin, pancreatin) for conditions like chronic
pancreatitis, cystic fibrosis, or after pancreatic surgery.
 • Wound Healing: Proteolytic enzymes to debride wounds and promote tissue repair.
• Thrombolytic Therapy: Enzymes to dissolve blood clots in myocardial infarction, pulmonary embolism, or deep vein
thrombosis.
• Genetic Enzyme Deficiencies: ERT for lysosomal storage diseases (e.g., Gaucher’s, Fabry’s, Pompe disease).
• Ophthalmology: Nucleases for viral eye infections; hyaluronidase for scar management.

32. Enzymodiagnostics. Enzymes as objects of laboratory research. Study of the isozyme spectrum. Enzymes as analytical
reagents.
Ans. Enzymodiagnostics: Principles, Laboratory Use, Isozyme Spectrum, and Enzymes as Analytical Reagents
Enzymodiagnostics: Principles and Significance
Enzymodiagnostics is the use of enzyme activity measurements in biological fluids (such as blood serum, plasma, urine) for the
diagnosis, monitoring, and prognosis of diseases. Because enzymes are highly specific to tissues and metabolic pathways,
changes in their concentration or activity in body fluids often reflect damage or dysfunction in specific organs or tissues1.
Key principles:
• Many enzymes are normally intracellular; their appearance or increased activity in blood indicates cell damage or
disease.
• The pattern and degree of enzyme elevation can help localize the affected organ and assess the severity and stage of
disease.
• Enzyme activity is measured using standardized laboratory assays, under optimal conditions of pH, temperature, and
substrate concentration1.
Enzymes as Objects of Laboratory Research
Enzymes are central to laboratory diagnostics for several reasons:
• Diagnostic markers: Enzyme activity in serum or plasma is used to detect and monitor diseases. For example:
• Myocardium: Creatine phosphokinase-MB (CPK-MB), lactate dehydrogenase (LDH)
• Liver: Alanine aminotransferase (ALT), aspartate aminotransferase (AST)
• Pancreas: Amylase, lipase
• Bone: Alkaline phosphatase
• Prostate, blood cells: Acid phosphatase
• Nerve tissue: Acetylcholinesterase
• Pathological changes: Metabolic disorders or tissue damage alter enzyme concentrations in biological fluids, making
enzyme assays indispensable for diagnosis and monitoring1.
• Types of enzymes in blood:
• Cellular enzymes: Released from damaged cells (organ-specific or non-specific).
• Secretory enzymes: Synthesized for physiological function in blood (e.g., coagulation enzymes).
• Excretory enzymes: Produced by glands and enter the blood (e.g., amylase, lipase).
Study of the Isozyme Spectrum
Isozymes (isoenzymes) are different molecular forms of the same enzyme that catalyze the same reaction but differ in amino
acid sequence, kinetic properties, and tissue distribution. Studying the isozyme spectrum is crucial in clinical diagnostics
because:
• Different tissues express characteristic isozyme patterns.
• Measuring specific isozymes allows precise localization of tissue damage.
Example: Lactate Dehydrogenase (LDH) Isozymes
• LDH is a tetramer made of two types of subunits (H and M), forming five isozymes:
• LDH1 (HHHH): Heart, erythrocytes
• LDH2 (HHHM): Heart, erythrocytes
• LDH3 (HHMM): Brain, kidney
• LDH4 (HMMM): Skeletal muscle, liver
• LDH5 (MMMM): Skeletal muscle, liver
• After myocardial infarction, LDH1 becomes predominant in serum, aiding in diagnosis1.
Other clinically important isozymes include:
• Creatine kinase (CK): CK-MB (heart), CK-BB (brain), CK-MM (muscle)
• Alkaline phosphatase (ALP): Liver, bone, intestinal, placental isoforms
Enzymes as Analytical Reagents
Enzymes are widely used as reagents in clinical chemistry for the quantitative determination of various substances in biological
samples. Their high specificity and catalytic efficiency make them ideal for analytical applications.
Examples of enzyme-based analytical assays:
• Glucose: Glucose oxidase–peroxidase method
• Urea: Urease-based assays
• Uric acid: Uricase (urate oxidase) method
• Cholesterol: Cholesterol oxidase method
• Ethanol: Alcohol dehydrogenase method
 • Triglycerides: Glycerol-3-phosphate oxidase method
• Lactic acid: Lactate oxidase method1
Enzyme-linked immunosorbent assay (ELISA):
• Uses enzymes as markers to detect antigens or antibodies in samples, widely used for infectious disease diagnosis,
hormone levels, tumor markers, and drug screening.
Methods for Enzyme Assay and Isozyme Detection
• Spectrophotometric assays: Measure changes in absorbance as substrate is converted to product.
• Electrophoresis: Separates isozymes based on charge and size for pattern analysis.
• Chromatography: Used for purification and identification of enzymes and isozymes.
• Immunochemical methods: Use antibodies specific to isozymes for detection.
Clinical Relevance
• Diagnosis: Enzyme activity profiles are essential for diagnosing myocardial infarction, hepatitis, pancreatitis, bone
diseases, muscle disorders, and certain cancers.
• Monitoring: Serial enzyme measurements monitor disease progression or response to therapy.
• Screening: Enzyme assays are used in newborn screening for metabolic disorders.

33. Immobilized enzymes. Properties. Applications.

Ans. Immobilized Enzymes: Properties and Applications
What Are Immobilized Enzymes?
Immobilized enzymes are enzymes that have been physically confined or localized in a certain defined region (usually on or
within a solid support), while retaining their catalytic activity and allowing them to be used repeatedly and continuously in
various processes4. Immobilization typically involves binding the enzyme to a carrier or trapping it within a matrix.
Methods of Immobilization
• Covalent immobilization: The enzyme is covalently bonded to a solid carrier, creating a strong and stable enzyme-carrier
complex.
• Non-covalent immobilization: The enzyme is adsorbed onto carriers, entrapped in gels, or encapsulated in
microcapsules. These interactions are weaker and may allow some enzyme dissociation over time4.
Properties of Immobilized Enzymes
• Reusability: Immobilized enzymes can be separated from the reaction mixture and reused multiple times, making
processes more economical.
• Stability: Immobilization often increases enzyme stability against changes in temperature, pH, and the presence of
inhibitors or organic solvents.
• Controlled reaction: The reaction can be easily started or stopped by adding or removing the immobilized enzyme.
• Continuous operation: Suitable for use in continuous-flow reactors, allowing for large-scale industrial processing.
• Reduced contamination: The enzyme does not mix with the product, reducing downstream purification steps.
• Potential for altered activity: Sometimes, immobilization can affect enzyme conformation, leading to changes in activity
or specificity.
Applications of Immobilized Enzymes
• Industrial biocatalysis: Used in the production of pharmaceuticals, chemicals, and food products (e.g., glucose
isomerase for high-fructose corn syrup production).
• Biosensors: Immobilized enzymes are key components in biosensors, such as glucose oxidase in blood glucose meters.
 • Medical devices: Incorporated into wound dressings and prostheses with thrombolytic (clot-dissolving) activity4.
• Waste treatment: Applied in reactors to degrade pollutants or toxins in wastewater.
• Analytical chemistry: Used as reagents in diagnostic kits and laboratory assays for the quantitative determination of
various substances.
• Extracorporeal reactors: Used in devices for blood purification (e.g., removal of toxins or drugs from blood outside the
body)4.
• Microencapsulation: Immobilized enzymes can be included in microcapsules for targeted drug delivery or controlled
release applications.
Examples
• Dressings for wound care and healing: Immobilized proteolytic enzymes help clean wounds and promote healing.
• Prostheses with thrombolytic enzymes: Reduce the risk of blood clots on artificial devices.
• Biosensors: Glucose oxidase immobilized on electrodes for continuous blood glucose monitoring in diabetic patients4.

34. Enzyme immunoassay, principle, application.

Ans. Principle
Enzyme immunoassay (EIA), also known as enzyme-linked immunosorbent assay (ELISA), is an immunological technique used
to detect and quantify specific antigens or antibodies in biological samples. The core principle relies on the highly specific
binding between an antigen and its corresponding antibody, combined with a sensitive enzymatic reaction that produces a
measurable signal.
How it works:
• An antigen (or antibody) is immobilized on a solid surface (typically a microtiter plate).
 • A specific antibody (or antigen) linked to an enzyme is added. This forms an antigen-antibody complex.
• After washing away unbound components, a substrate for the enzyme is introduced.
• The enzyme catalyzes a reaction with the substrate, resulting in a color change, fluorescence, or luminescence.
• The intensity of the signal is proportional to the amount of antigen or antibody present in the sample and can be
measured using a spectrophotometer or similar device.
Types of ELISA/EIA:
• Direct ELISA: Detects antigen with an enzyme-labeled antibody.
• Indirect ELISA: Uses a secondary enzyme-labeled antibody to detect the primary antibody bound to the antigen.
• Sandwich ELISA: Captures antigen between two antibodies (capture and detection antibodies), both specific for
different epitopes.
• Competitive ELISA: Measures the competition between labeled and unlabeled antigen for antibody binding.
Application in Medicine
Enzyme immunoassay is widely used in clinical diagnostics, research, and public health due to its sensitivity, specificity, and
versatility. Key applications include:
• Detection and quantification of hormones:
Used to measure levels of hormones such as luteinizing hormone, prolactin, testosterone, and human chorionic
gonadotropin (hCG).
• Diagnosis of infectious diseases:
Detects antibodies or antigens related to infections such as HIV, hepatitis A/B/C, influenza, and COVID-19.
• Autoimmune disease testing:
Identifies autoantibodies (e.g., anti-dsDNA, ANA) in conditions like systemic lupus erythematosus.
 • Tumor marker measurement:
Quantifies tumor markers such as prostate-specific antigen (PSA) and carcinoembryonic antigen (CEA) for cancer
diagnosis and monitoring.
• Screening of blood donations:
Ensures safety by detecting viral antigens or antibodies (e.g., anti-HIV, anti-HCV, HBsAg) in donor blood.
• Drug testing:
Used for detecting drugs of abuse (e.g., amphetamines, methamphetamines, cocaine) in biological fluids.
• Epidemiology and outbreak tracking:
Helps monitor disease outbreaks by detecting antibodies indicating past exposure (e.g., cholera, Lyme disease).

35. Vitamins. Common signs. Classification. Hypervitaminosis, hypovitaminosis. Antivitamins. Exogenous and endogenous
causes of vitamin deficiency.
Ans. Common Signs and General Features of Vitamins
• Definition: Vitamins are a group of low-molecular-weight organic compounds essential for normal physiological
functions, growth, survival, and reproduction. They are not synthesized in sufficient quantities by the human body (or
not at all) and must be obtained from the diet1.
• Function: Vitamins are required for biochemical reactions, often as coenzymes, antioxidants, or regulators of gene
expression. Their absence leads to characteristic deficiency diseases.
• Families: Many vitamins are not single substances but families of chemically related compounds (vitamers) with similar
biological activity1.
• Detection: The importance of vitamins is often revealed not by their presence, but by the symptoms that arise from
their absence1.
Classification of Vitamins
By Solubility:
• Fat-soluble vitamins: A (retinol), D (cholecalciferol), E (tocopherol), K (phylloquinone). These are stored in body tissues
and can accumulate to toxic levels1.
• Water-soluble vitamins: C (ascorbic acid), B-complex group (B1-thiamine, B2-riboflavin, B3-niacin, B5-pantothenic acid,
B6-pyridoxine, B7-biotin, B9-folate, B12-cobalamin). These are not stored in large amounts and excess is usually
excreted13.
By Function:
• Coenzyme vitamins: Most B vitamins and vitamin C act as coenzymes or their precursors.
• Prohormone vitamins: Vitamin D acts as a hormone after activation.
• Antioxidant vitamins: Vitamins E and C.
• Gene expression regulators: Vitamins A and D1.
Hypervitaminosis
• Definition: Hypervitaminosis is a condition caused by excessive intake and storage of vitamins, leading to toxicity and
adverse health effects1.
• Most Common: Fat-soluble vitamins (A and D) are most often responsible, as they are stored in the body and not readily
excreted.
• Symptoms: May include irritability, headache, skin changes, bone pain, liver damage, and in severe cases, organ failure.
For example, hypervitaminosis A can cause neurological symptoms, skin peeling, and bone changes; hypervitaminosis D
can lead to hypercalcemia and tissue calcification1.
Hypovitaminosis
 • Definition: Hypovitaminosis is a disorder caused by a deficiency of one or more vitamins, which may result from
inadequate intake, absorption, increased requirements, or increased losses1.
• Symptoms: Vary depending on the vitamin, but can include anemia (B12, folate), dermatitis (B2, B3, B6), night blindness
(A), scurvy (C), rickets/osteomalacia (D), and bleeding disorders (K)13.
• Avitaminosis: Complete absence of a vitamin, leading to the most severe form of deficiency disease1.
Antivitamins
• Definition: Antivitamins are chemical compounds that inhibit the absorption or biological action of vitamins13.
• Mechanisms:
• Structural analogs that compete with the vitamin for absorption or enzyme binding.
• Compounds that chemically modify the vitamin, making it inactive or poorly absorbed.
• Enzymes or proteins that bind or degrade vitamins, reducing their physiological effects13.
• Examples: Methotrexate (antifolate), dicumarol (antagonist of vitamin K), pyrithiamine (antithiamine), avidin (binds and
inactivates biotin)3.
Exogenous and Endogenous Causes of Vitamin Deficiency
Exogenous Causes:
• Poor dietary intake: Monotonous or unbalanced diets, low in micronutrients, especially animal-source foods.
• Malnutrition: Protein-energy malnutrition reduces vitamin intake and absorption.
• Food shortages or seasonal variation: Limited access to diverse foods.
• Social and economic factors: Poverty, illiteracy, low education.
 • Increased physiological needs: Growth, pregnancy, lactation, infections, or chronic diseases can raise vitamin
requirements1.
Endogenous Causes:
• Malabsorption: Disorders of the gastrointestinal tract, chronic diarrhea, or diseases affecting absorption (e.g., celiac
disease, Crohn's disease).
• Lack of synthesis: Failure of intestinal bacteria to synthesize certain vitamins (e.g., vitamin K, some B vitamins).
• Metabolic disorders: Enzymopathies affecting vitamin activation or utilization.
• Increased losses: Due to renal disease, parasitic infections, or increased excretion.
• Drug interactions: Use of antivitamins, certain medications, or toxins that interfere with vitamin metabolism13.

36. Water-soluble and fat-soluble vitamins. Their biotransformation in the body. Coenzyme functions of vitamins. Examples.
Ans. Water-Soluble and Fat-Soluble Vitamins
Water-Soluble Vitamins:
These vitamins dissolve in water, are not stored in large amounts in the body, and excess is excreted in urine. They must be
regularly supplied in the diet. The main water-soluble vitamins are:
• Vitamin C (ascorbic acid)
• B-complex vitamins: B1 (thiamine), B2 (riboflavin), B3 (niacin), B5 (pantothenic acid), B6 (pyridoxine), B7 (biotin), B9
(folic acid), B12 (cobalamin)23.
Fat-Soluble Vitamins:
These vitamins dissolve in fats and oils, are absorbed along with dietary fat, and can be stored in body tissues (mainly the liver
and adipose tissue). The main fat-soluble vitamins are:
 • Vitamin A (retinol)
• Vitamin D (cholecalciferol)
• Vitamin E (tocopherol)
• Vitamin K (phylloquinone)3.
Biotransformation of Vitamins in the Body
Water-Soluble Vitamins:
• Absorbed directly into the blood from the intestine.
• Not extensively stored; excess is excreted in urine.
• Many are converted into metabolically active coenzyme forms after absorption23.
Fat-Soluble Vitamins:
• Absorbed with dietary fats via the lymphatic system.
• Require bile salts for absorption.
• Stored in the liver and adipose tissue; released as needed.
• Often require metabolic activation (e.g., vitamin D is hydroxylated in the liver and kidney to its active form; vitamin A is
converted to retinal and retinoic acid)3.
Coenzyme Functions of Vitamins
Many vitamins act as coenzymes or are precursors for coenzymes, which are essential for enzyme-catalyzed reactions in
metabolism.
Examples of Water-Soluble Vitamins and Their Coenzyme Functions:
 • Vitamin B1 (Thiamine):
Converted to thiamine pyrophosphate (TPP), a coenzyme for decarboxylation of α-keto acids (e.g., pyruvate
dehydrogenase, transketolase in the pentose phosphate pathway)2.
• Vitamin B2 (Riboflavin):
Precursor of FMN and FAD, which are coenzymes for oxidation-reduction (redox) reactions (e.g., succinate
dehydrogenase in the TCA cycle)25.
• Vitamin B3 (Niacin):
Precursor of NAD and NADP, coenzymes for redox reactions in glycolysis, TCA cycle, and oxidative phosphorylation25.
• Vitamin B5 (Pantothenic acid):
Component of coenzyme A (CoA), essential for acyl-group transfer in metabolism (e.g., acetyl-CoA formation)2.
• Vitamin B6 (Pyridoxine):
Converted to pyridoxal phosphate (PLP), a coenzyme for amino acid metabolism (transamination, decarboxylation,
glycogen phosphorylase)2.
• Vitamin B7 (Biotin):
Coenzyme for carboxylation reactions (e.g., pyruvate carboxylase, acetyl-CoA carboxylase)2.
• Vitamin B9 (Folic acid):
Converted to tetrahydrofolate (THF), a coenzyme for one-carbon transfer in nucleotide and amino acid metabolism2.
• Vitamin B12 (Cobalamin):
Coenzyme for methylation reactions and isomerization of methylmalonyl-CoA to succinyl-CoA2.
• Vitamin C (Ascorbic acid):
Coenzyme for hydroxylation reactions (collagen synthesis), antioxidant, enhances iron absorption23.
Examples of Fat-Soluble Vitamins and Their Functions:
 • Vitamin A:
Retinal is a cofactor for vision (component of rhodopsin); retinoic acid regulates gene expression for growth and
differentiation3.
• Vitamin D:
After activation (hydroxylation in liver and kidney), acts as a hormone to regulate calcium and phosphate metabolism3.
• Vitamin E:
Functions as an antioxidant, protecting cell membranes from oxidative damage3.
• Vitamin K:
Coenzyme for γ-carboxylation of glutamic acid residues in clotting factors, essential for blood coagulation3.

37. Vitamin A. Chemical structure. Sources, provitamins, need. Signs of insufficiency and hypervitaminosis, biological role.
Ans. Chemical Structure
Vitamin A refers to a group of fat-soluble compounds including:
• Retinol (alcohol form, main storage and transport form)
• Retinal (aldehyde form, essential for vision)
• Retinoic acid (acid form, regulates gene expression)
• Retinyl esters (storage form in the liver)
The basic structure is a cyclohexenyl ring with an isoprenoid side chain. Retinol (C20H30O) is the most biologically active
form2.
Provitamins
 • Provitamin A are carotenoids, the most important being β-carotene and other carotenoids found in plants. These are
cleaved in the intestinal mucosa to form retinol.
• β-carotene is the most efficient provitamin A; 1 molecule yields 2 molecules of vitamin A2.
Dietary Sources
Animal sources (preformed vitamin A):
• Liver (beef, pork, fish, poultry)
• Fish liver oils (cod liver oil)
• Eggs (yolk)
• Dairy products (butter, cheese, whole milk)
Plant sources (provitamin A carotenoids):
• Carrots
• Sweet potatoes
• Spinach
• Kale
• Broccoli
• Pumpkin
• Leafy green and yellow/orange vegetables2.
Daily Requirement
• Recommended Dietary Allowance (RDA):
• Adult males: 900 µg retinol activity equivalents (RAE)/day
 • Adult females: 700 µg RAE/day
• Increased needs during pregnancy and lactation2.
Biological Role
• Vision: Retinal is a component of rhodopsin in the retina, essential for low-light (scotopic) vision. Light converts 11-cis-
retinal to all-trans-retinal, triggering a nerve impulse.
• Gene transcription: Retinoic acid regulates gene expression by binding to nuclear receptors, affecting cell growth,
differentiation, and embryonic development.
• Epithelial integrity: Maintains normal structure and function of epithelial tissues (skin, mucous membranes, respiratory,
urinary, and reproductive tracts).
• Immune function: Supports T-cell differentiation and proliferation, enhancing immune response.
• Reproduction and growth: Essential for normal reproduction and fetal development.
• Bone metabolism: Involved in bone remodeling via effects on osteoblasts and osteoclasts2.
Signs of Vitamin A Insufficiency (Hypovitaminosis)
• Night blindness (nyctalopia): Early symptom due to impaired rhodopsin regeneration.
• Xerophthalmia: Dryness of conjunctiva and cornea, progressing to Bitot's spots, keratomalacia (corneal ulceration), and
possible blindness.
• Follicular hyperkeratosis: Rough, dry skin due to abnormal keratinization.
• Impaired immunity: Increased susceptibility to infections.
• Growth retardation: In children, due to impaired cell differentiation.
• Abnormal bone growth and reproductive issues2.
Signs of Hypervitaminosis A (Excess)
• Acute toxicity: Nausea, vomiting, headache, dizziness, blurred vision, increased intracranial pressure.
• Chronic toxicity: Skin changes (dryness, peeling, itching), hair loss, cracking at mouth corners, bone pain, liver
enlargement, irritability, drowsiness, double vision, and in severe cases, liver damage and increased risk of fractures.
• Teratogenic effects: High doses during pregnancy can cause birth defects.
• Altered metabolism of other fat-soluble vitamins (D, E, K)2.

38. Vitamin D. Chemical structure. Exogenous and endogenous sources, need, transformation in the body. Causes of hyper-
and hypovitaminosis, manifestations. Biological role.
Ans. Chemical Structure
Vitamin D refers to a group of fat-soluble secosteroids. The two major forms are:
• Vitamin D2 (ergocalciferol): Derived from plant sources and fungi.
• Vitamin D3 (cholecalciferol): Synthesized in animal skin and found in animal-based foods.
Both forms share a similar steroid backbone but differ in their side chains. The active hormonal form is 1,25-
dihydroxycholecalciferol (calcitriol), which is produced in the body through sequential hydroxylation.
Exogenous and Endogenous Sources
Exogenous (Dietary) Sources:
• Animal sources: Fatty fish (salmon, mackerel, sardines, tuna), fish liver oils, egg yolk, liver, and fortified foods (milk,
cereals).
• Plant sources: Some mushrooms (exposed to UV light) provide vitamin D2.
 • Supplements: Available as D2 or D3.
Endogenous (Internal) Source:
• Skin synthesis: The main natural source. 7-dehydrocholesterol in the skin is converted to vitamin D3 (cholecalciferol)
upon exposure to ultraviolet B (UVB) rays from sunlight1.
Physiological Requirement
• Recommended Dietary Allowance (RDA):
• Infants (0–12 months): 400 IU (10 µg)/day
• Children and adults (1–70 years): 600 IU (15 µg)/day
• Adults >70 years: 800 IU (20 µg)/day
• Pregnant/lactating women: 600 IU (15 µg)/day1
Requirements may be higher in individuals with limited sun exposure, darker skin, or malabsorption syndromes.
Transformation in the Body
1. Synthesis/Intake:
• Vitamin D3 is synthesized in the skin or obtained from animal foods; D2 from plant foods.
2. Liver Hydroxylation:
• Both D2 and D3 are transported to the liver, where they are hydroxylated to 25-hydroxycholecalciferol (25(OH)D),
the main circulating form.
3. Kidney Hydroxylation:
• 25(OH)D is further hydroxylated in the kidney to 1,25-dihydroxycholecalciferol (1,25(OH)2D, calcitriol), the
biologically active hormone.
 4. Action:
• Calcitriol binds to the vitamin D receptor (VDR) in target tissues, regulating gene transcription and
calcium/phosphate metabolism1.
Causes and Manifestations of Hypovitaminosis (Deficiency)
Causes:
• Insufficient sunlight exposure (geographic, cultural, or lifestyle factors)
• Poor dietary intake (vegan diets, lack of fortified foods)
• Malabsorption (intestinal diseases)
• Chronic kidney or liver disease (impaired hydroxylation)
• Increased physiological needs (pregnancy, growth, elderly)
Manifestations:
• Children: Rickets (impaired bone mineralization, bone deformities like bowed legs)
• Adults: Osteomalacia (bone pain, muscle weakness, increased fracture risk), osteoporosis
• Other: Increased risk of infections, possible links to cardiovascular disease, autoimmune disorders, and some cancers1.
Causes and Manifestations of Hypervitaminosis (Excess)
Causes:
• Excessive supplementation (rare from diet or sun exposure)
• Disorders causing increased sensitivity to vitamin D
Manifestations:
• Hypercalcemia: High blood calcium, leading to nausea, vomiting, constipation, dehydration, muscle weakness, confusion
 • Calcification: Overcalcification of bones, soft tissues, kidneys (nephrocalcinosis), and heart
• Other: Hypertension, decreased appetite, irritability, fatigue1.
Biological Role
• Calcium and Phosphate Homeostasis:
• Increases intestinal absorption and renal reabsorption of calcium and phosphate.
• Stimulates synthesis of proteins (e.g., calbindin) involved in calcium transport.
• Bone Health:
• Stimulates bone remodeling, induces osteocalcin expression, and suppresses type I collagen synthesis.
• Essential for normal bone growth and maintenance.
• Cellular Growth and Differentiation:
• Regulates growth and differentiation of many cell types, including immune cells.
• Immune Function:
• Modulates immune response; many immune cells express VDR and can synthesize active vitamin D1.
• Other Roles:
• May play a role in prevention of certain chronic diseases (diabetes, cardiovascular disease, multiple sclerosis,
some cancers).

39. Vitamin E. Chemical structure. Sources, daily requirement. Deficiency signs, biological role.
Ans. Chemical Structure
Vitamin E is the collective name for a group of fat-soluble compounds called tocopherols and tocotrienols. The most
biologically active and common form in humans is α-tocopherol.
• Core structure: Chromanol ring system (chroman) with a long saturated phytyl side chain (in tocopherols) or an
unsaturated side chain (in tocotrienols).
• Forms: α-, β-, γ-, and δ-tocopherol (differ in the number and position of methyl groups on the chromanol ring).
α-Tocopherol is the principal form found in the human body and used in supplementation1.
Sources
Rich dietary sources of vitamin E include:
• Vegetable oils: Wheat germ oil (very high), sunflower oil, safflower oil, canola (rapeseed) oil, hazelnut oil, almond oil,
grapeseed oil.
• Nuts and seeds: Almonds, sunflower seeds, hazelnuts.
• Whole grains: Wheat germ.
• Green leafy vegetables: Spinach, kale.
• Other: Almond butter, sunflower seed kernels.
Animal products are generally poor sources of vitamin E1.
Daily Requirement
• Recommended Dietary Allowance (RDA):
• Adults (≥19 years): 15 mg α-tocopherol/day
• Pregnancy: 15 mg/day
• Lactation: 19 mg/day
 • Children: 6–15 mg/day, depending on age
1 International Unit (IU) of vitamin E is equivalent to about 0.667 mg d-α-tocopherol or 0.9 mg dl-α-tocopherol1.
Deficiency Signs
Vitamin E deficiency is rare but can occur in:
• Individuals with fat malabsorption syndromes (e.g., cystic fibrosis, chronic pancreatitis, cholestatic liver disease)
• Premature infants with low birth weight
Clinical signs and consequences:
• Neuromuscular problems: Muscle weakness, ataxia (impaired coordination), peripheral neuropathy
• Hemolytic anemia: Increased fragility and destruction of red blood cells
• Retinal degeneration: Visual disturbances, retinitis pigmentosa
• Impaired immune function
• Infertility: Impaired fertility in both sexes (the basis for the name "antisterility vitamin")
• Other: Dry skin, hair loss, leg cramps, increased risk of degenerative diseases (e.g., cancer, atherosclerosis, chronic
inflammation, cataracts)1.
Biological Role
• Antioxidant activity:
• Vitamin E is a major chain-breaking antioxidant in cell membranes. It scavenges and neutralizes free radicals
(reactive oxygen species), especially preventing lipid peroxidation in polyunsaturated fatty acids of membrane
phospholipids13.
• Membrane protection:
 • Maintains the integrity and stability of biological membranes.
• Synergy with vitamin A:
• Increases biological activity of vitamin A.
• Heme synthesis:
• Stimulates the synthesis of heme-containing proteins (hemoglobin, myoglobin, cytochromes, catalase,
peroxidase).
• Other roles:
• Supports immune function, protects against oxidative stress-related diseases, may reduce risk of some chronic
diseases.

40. Vitamin K. Chemical structure. Sources, need. Role in providing hemostasis. Antivitamins as medicines.
Ans. Chemical Structure
Vitamin K refers to a family of fat-soluble compounds with a common chemical structure: a 2-methyl-1,4-naphthoquinone ring.
The main forms are:
• Phylloquinone (Vitamin K1): Found in plants; has a phytyl side chain.
• Menaquinones (Vitamin K2): Produced by intestinal bacteria; have multiple isoprenoid side chains.
• Menadione (Vitamin K3): A synthetic, water-soluble form.
Sources
• Vitamin K1 (phylloquinone): Chiefly from green leafy vegetables (spinach, kale, broccoli, lettuce), and some plant oils.
 • Vitamin K2 (menaquinones): Synthesized by gut bacteria; also found in animal products (meat, eggs, cheese)
and fermented foods (e.g., natto).
• Menadione (K3): Synthetic, used in some supplements and animal feeds.
Note: The chief source of vitamin K for most people is synthesis by bacteria in the large intestine, though dietary intake is
also important1.
Daily Requirement
• Adequate Intake (AI):
• Adult men: 120 µg/day
• Adult women: 90 µg/day
• Pregnancy/lactation: 90 µg/day
Role in Providing Hemostasis
Vitamin K is essential for normal blood clotting (hemostasis). Its main function is as a coenzyme for the enzyme gamma-
glutamyl carboxylase, which catalyzes the carboxylation of specific glutamic acid residues on several key proteins involved in
coagulation. This process produces gamma-carboxyglutamic acid (Gla) residues, enabling these proteins to bind calcium ions,
which is critical for their biological activity.
Vitamin K-dependent coagulation factors:
• Factor II (prothrombin)
• Factor VII
• Factor IX
• Factor X
• Proteins C, S, and Z (anticoagulant proteins)
Mechanism:
• Vitamin K is converted to its active hydroquinone form (KH2), which donates electrons for the carboxylation reaction.
• After carboxylation, vitamin K is oxidized to vitamin K epoxide, which is then recycled back to its active form by vitamin K
epoxide reductase (the vitamin K cycle).
• This cycle is the target of certain anticoagulant drugs1.
Deficiency:
Leads to impaired synthesis of clotting factors, resulting in bleeding disorders-manifested as easy bruising, petechiae,
hematomas, excessive bleeding after injury or surgery, and in severe cases, life-threatening hemorrhage. In infants, deficiency
can cause hemorrhagic disease of the newborn.
Antivitamins as Medicines
Antivitamins are compounds that antagonize the action of vitamins. For vitamin K, the most clinically important antivitamins
are coumarin derivatives:
• Warfarin and dicumarol are vitamin K antagonists widely used as oral anticoagulants.
• Mechanism: They inhibit vitamin K epoxide reductase, blocking the recycling of vitamin K to its active form and
thus preventing the gamma-carboxylation of clotting factors. This reduces the synthesis of functional coagulation
proteins, resulting in anticoagulation14.
• Clinical use: Prevention and treatment of thromboembolic disorders (e.g., deep vein thrombosis, pulmonary embolism,
atrial fibrillation, prosthetic heart valves).
Note: Overdose or excessive inhibition can cause dangerous bleeding; vitamin K is used as an antidote for warfarin overdose.

41. Vitamin B1. Chemical structure. Sources, need, coherent forms. Biological role.
Ans. Chemical Structure
Vitamin B1, or thiamine, is a water-soluble vitamin composed of a pyrimidine ring and a thiazole ring linked by a methylene
bridge. Its chemical formula is C12H17N4OS.
• The biologically active coenzyme form is thiamine pyrophosphate (TPP), also called thiamine diphosphate, which is
formed by the addition of two phosphate groups to thiamine1.
Sources
Plant sources:
• Outer layers and germ of cereal grains (rice polishings, wheat germ, bran)
• Whole grains, peas, beans, nuts, prunes
Animal sources:
• Liver, pork, other meats, eggs
Note: Thiamine is sensitive to heat and can be lost during cooking and processing, especially in foods made from refined
grains1.
Daily Requirement
• Adults: Minimum requirement is 1.0 mg/day, or about 0.5 mg per 1000 kcal of energy intake
• Children: Ranges from 0.4 mg/day (infants) to 1.3 mg/day (preadolescents, 10–12 years)
• Requirement increases with high carbohydrate intake, pregnancy, lactation, and certain illnesses1.
Coenzyme (Metabolically Active) Forms
• Thiamine pyrophosphate (TPP): The main coenzyme form, also known as thiamine diphosphate
• Other forms: Thiamine, thiamine disulfide, thiamine hydrochloride, thiamine mononitrate (dietary forms)1
Biological Role
1. Coenzyme Function:
Thiamine pyrophosphate (TPP) acts as a coenzyme in several key metabolic reactions, especially in carbohydrate and amino
acid metabolism:
• Pyruvate dehydrogenase: Converts pyruvate to acetyl-CoA, linking glycolysis to the citric acid cycle
• α-Ketoglutarate dehydrogenase: Part of the citric acid cycle
• Branched-chain α-keto acid dehydrogenase: Metabolism of branched-chain amino acids
• Transketolase: In the pentose phosphate pathway, important for nucleotide synthesis and NADPH production12
2. Importance in Energy Metabolism:
Thiamine is essential for the oxidative decarboxylation of α-keto acids and for the transfer of two-carbon units, making it vital
for energy production from carbohydrates and for the functioning of the nervous system12.
3. Nervous System Function:
Thiamine is crucial for nerve conduction and neurotransmitter synthesis, explaining why deficiency primarily affects the
nervous and cardiovascular systems1.

42. Vitamin B2. Chemical structure. Sources, need. Coenzyme forms. Biological role.
Ans. Chemical Structure
Vitamin B2, also known as riboflavin, is a water-soluble vitamin with a structure consisting of an isoalloxazine ring (flavin)
attached to a ribitol (sugar alcohol) side chain.
• The flavin ring system is responsible for the vitamin’s characteristic yellow color.
• Riboflavin is the precursor for two important coenzymes: flavin mononucleotide (FMN) and flavin adenine dinucleotide
(FAD)1.
Sources
Rich dietary sources include:
• Milk and dairy products (major source in many diets)
• Eggs
• Cheese
• Liver, kidneys, lean meats
• Leafy green vegetables
• Legumes
• Mushrooms
• Almonds
• Whole and enriched grains
Riboflavin is widely distributed in both animal and plant foods, but it is sensitive to light and can be destroyed by exposure to
sunlight1.
Daily Requirement
• Adults: 1.5 to 1.8 mg/day
• Women in late pregnancy: 2.0 mg/day
• During lactation: 2.5 mg/day
• Children: 1.0 to 1.8 mg/day
• Infants: 0.6 mg/day
• Adolescents: 2.0 to 2.5 mg/day
Requirements increase with growth, pregnancy, lactation, and increased energy expenditure1.
Coenzyme Forms
Riboflavin is converted in the body to two metabolically active coenzymes:
• Flavin mononucleotide (FMN)
• Flavin adenine dinucleotide (FAD)
These coenzymes are collectively known as flavoproteins when bound to enzymes1.
Biological Role
1. Redox Reactions:
FMN and FAD act as coenzymes for a wide variety of enzymes (flavoproteins) that catalyze oxidation-reduction (hydrogen
transfer) reactions in metabolism. They are essential for:
• Electron transport chain (respiratory chain): FAD is a coenzyme for succinate dehydrogenase (Complex II).
• Fatty acid oxidation: FAD is a coenzyme for acyl-CoA dehydrogenase.
• Amino acid oxidation: FAD and FMN are coenzymes for various amino acid oxidases.
2. Energy Production:
Riboflavin is crucial for the metabolism of carbohydrates, fats, and proteins, as it is required for the function of enzymes in the
citric acid cycle and electron transport chain, both of which generate ATP14.
3. Other Functions:
• Involved in the fixation of glycogen in the liver.
• Contributes to the maintenance of normal vision and healthy skin.
• FMN is a coenzyme for cytochrome c reductase and L-amino acid oxidase.
• FAD is a coenzyme for xanthine oxidase, aldehyde oxidase, glycine oxidase, diaphorase, and others1.
Deficiency Signs:
Riboflavin deficiency impairs biological oxidation, especially in aerobic tissues. Clinical manifestations include:
• Lesions of the mouth (cheilosis), tongue (glossitis), angular stomatitis, and skin (seborrheic dermatitis)
• Oily, scaly skin rashes, especially around the nose, scrotum, vulva, and lips
• Eye symptoms (photophobia, itching, burning)
• Weakness, lassitude
• Anemia, often associated with other B vitamin deficiencies1.
43. Vitamin B3. Chemical structure. Sources, need, coenzyme forms, biological role.
Ans. Vitamin B3 (Niacin): Chemical Structure, Sources, Need, Coenzyme Forms, Biological Role
Chemical Structure
Vitamin B3 refers to two related compounds:
• Nicotinic acid (niacin): Pyridine-3-carboxylic acid
• Nicotinamide (niacinamide): Pyridine-3-carboxamide
Both share a pyridine ring structure and are water-soluble. They are collectively known as niacin and have equivalent vitamin
activity in the body2.
Sources
Animal sources:
• Liver
• Kidney
• Meat
 • Fish
Plant sources:
• Legumes (peas, beans, lentils)
• Nuts
• Certain green vegetables
• Coffee and tea
• Germ and pericarp (bran) of cereal grains
• Yeast
Note: Fruits, milk, and eggs are poor sources. Niacin can also be synthesized in the body from the essential amino acid
tryptophan, though this pathway is relatively inefficient2.
Daily Requirement
• Adults: 17–21 mg/day
• Infants: 6 mg/day
• Pre-adolescents: 17 mg/day
Requirements may increase during periods of growth, pregnancy, and lactation2.
Coenzyme Forms
Niacin is the precursor for two major coenzymes:
• Nicotinamide adenine dinucleotide (NAD)
• Nicotinamide adenine dinucleotide phosphate (NADP)
Both coenzymes are involved in reversible oxidation-reduction (redox) reactions, acting as hydrogen and electron carriers in
numerous metabolic pathways25.
Biological Role
1. Redox Reactions:
NAD and NADP function as coenzymes for many dehydrogenases and oxidoreductases, facilitating the transfer of hydrogen and
electrons. They are essential in:
• Glycolysis
• Citric acid (Krebs) cycle
• Oxidative phosphorylation
• Pentose phosphate pathway
• Fatty acid and amino acid metabolism25
2. NADP:
Especially important for biosynthetic (anabolic) reactions, such as fatty acid and cholesterol synthesis, and for maintaining
antioxidant systems (e.g., regeneration of reduced glutathione)25.
3. ADP-ribosylation:
NAD is a source of ADP-ribose for post-translational modification of proteins and for poly-ADP ribosylation, which is involved in
DNA repair mechanisms2.
4. Clinical Aspects:
• Deficiency: Causes pellagra, characterized by the "three Ds": Dermatitis, Diarrhea, and Dementia. Other symptoms
include skin lesions, gastrointestinal disturbances, and neuropsychiatric symptoms. Pellagra is most common in
populations relying heavily on untreated maize as a staple, as it is low in both niacin and tryptophan2.
 • Therapeutic use: Pharmacological doses of niacin are used to lower plasma cholesterol and triglyceride levels, but
excessive doses can cause toxicity (flushing, skin irritation, liver damage)2.

44. Biotin. Chemical structure, sources, need. Biological role.

Ans. Biotin (Vitamin B7): Chemical Structure, Sources, Requirement, and Biological Role
Chemical Structure
Biotin is a water-soluble B-complex vitamin, also known as vitamin B7 or vitamin H.
• Chemical structure: Biotin consists of a fused imidazolidone (ureido) ring and thiophene ring with a valeric acid side
chain.
• Molecular formula: C10H16N2O3S
• Active form: d-biotin
• Metabolically active form: Biocytin (biotin covalently bound to lysine) is the form in which biotin is incorporated into
enzymes1.
Sources
Dietary sources of biotin include:
• Animal sources: Liver, kidney, milk and milk products, egg yolk.
• Plant sources: Legumes, nuts, vegetables, grains, molasses.
• Others: Biotin is also synthesized by the normal bacterial flora of the intestine, which is a significant source for
humans12.
Daily Requirement
 • Adults: 25–50 µg/day
• Infants: 10–15 µg/day
• Children: 20–40 µg/day
• Requirement increases during pregnancy and lactation1.
Biological Role
• Coenzyme function:
Biotin acts as a coenzyme for several carboxylase enzymes that catalyze important carboxylation reactions (fixation of
CO₂) in metabolism:
• Acetyl-CoA carboxylase: Conversion of acetyl-CoA to malonyl-CoA (first step in fatty acid synthesis).
• Pyruvate carboxylase: Conversion of pyruvate to oxaloacetate (important in gluconeogenesis).
• Propionyl-CoA carboxylase: Conversion of propionyl-CoA to methylmalonyl-CoA (important in amino acid and
odd-chain fatty acid metabolism).
• β-methylcrotonyl-CoA carboxylase: Involved in leucine metabolism1.
• Other roles:
• Participates in deamination and decarboxylation of amino acids such as aspartic acid, serine, and threonine.
• Involved in the synthesis of fatty acids, gluconeogenesis, and amino acid catabolism.
Deficiency
• Causes:
• Rare, but may occur with prolonged consumption of raw egg whites (which contain avidin, a protein that binds
biotin and prevents its absorption), long-term antibiotic therapy, or genetic defects in biotinidase or
holocarboxylase synthase.
 • Symptoms:
• Dermatitis (especially of the extremities), pallor of skin and mucous membranes, anorexia, nausea, muscle pain,
hyperesthesia, depression, lassitude, somnolence, anemia, hypercholesterolemia, hair loss (alopecia), and
incoordination of movements1.
• In infants, can cause a rare condition called Leiner’s disease (exfoliative dermatitis).

45. Vitamin B6. Chemical structure. Sources, need, coenzyme forms, biological role.
Ans. Vitamin B6: Chemical Structure, Sources, Need, Coenzyme Forms, Biological Role
Chemical Structure
Vitamin B6 is a family of water-soluble compounds with similar activity, collectively called vitamers:
• Pyridoxine (pyridoxol)
• Pyridoxal
• Pyridoxamine
All share a pyridine ring with different functional groups:
• Pyridoxine: CH₂OH group
• Pyridoxal: CHO group
• Pyridoxamine: CH₂NH₂ group
The most important metabolically active form is pyridoxal 5-phosphate (PLP), which is formed by phosphorylation of the
vitamin in the body2.
Sources
Rich sources of vitamin B6 include:
• Animal sources: Liver, kidney, meat, fish
• Plant sources: Legumes (peas, beans, lentils), nuts, whole grains, certain green vegetables, coffee, tea
• Other: Yeast, germ and pericarp (bran) of cereal grains
Fruits, milk, and eggs are poor sources of vitamin B62.
Daily Requirement
• Adults: 2 mg/day
• Infants: 0.3–0.4 mg/day
• During pregnancy (second half): 2.5 mg/day
• Requirement increases in patients receiving certain medications (e.g., isoniazid for tuberculosis), during pregnancy, and
lactation2.
Coenzyme Forms
• Pyridoxal 5-phosphate (PLP): The main active coenzyme form
• Pyridoxamine 5-phosphate: Another active form
These coenzymes are involved in numerous enzymatic reactions, especially those related to amino acid metabolism2.
Biological Role
1. Amino Acid Metabolism:
PLP is a coenzyme for enzymes catalyzing:
• Transamination: Transfer of amino groups (essential for synthesis and breakdown of amino acids)
• Decarboxylation: Formation of neurotransmitters (serotonin, dopamine, GABA, norepinephrine, epinephrine)
 • Non-oxidative deamination
• Transsulfuration: Synthesis of cysteine from methionine
2. Neurotransmitter Synthesis:
PLP is essential for the biosynthesis of neurotransmitters, including serotonin, dopamine, GABA, and others. Deficiency can
result in neurological symptoms such as somnolence, confusion, and neuropathy2.
3. Glucose Metabolism:
PLP is a required coenzyme for glycogen phosphorylase, which is involved in glycogen breakdown.
4. Lipid Metabolism:
PLP is necessary for the biosynthesis of sphingolipids, important components of nerve tissue.
5. Hemoglobin Synthesis:
PLP is required for the synthesis and function of hemoglobin. It is involved in the reaction that produces δ-aminolevulinic acid,
a precursor of heme.
6. Gene Expression:
PLP influences gene expression, including decreasing transcription of certain genes by glucocorticoids2.
Deficiency Signs
• Dermatitis (seborrheic, skin rashes)
• Glossitis (sore, glossy, cracked tongue)
• Angular cheilitis (cracks at the corners of the mouth)
• Conjunctivitis
• Neurological symptoms: Somnolence, confusion, neuropathy, epileptiform convulsions in infants (due to impaired GABA
synthesis)
• Microcytic anemia: Due to impaired hemoglobin synthesis2
Mild deficiency may occur in women taking oral contraceptives or in patients on isoniazid therapy (which increases B6
requirement).
Therapeutic Uses
• Treatment of nausea and vomiting in pregnancy (morning sickness)
• Radiation sickness
• Muscular dystrophies
• Hyperoxaluria and prevention of recurrent oxalate kidney stones

46. Folic acid, chemical structure, sources, need, coenzyme forms, biological role.
Ans. Folic Acid (Vitamin B9): Chemical Structure, Sources, Requirement, Coenzyme Forms, Biological Role
Chemical Structure
Folic acid, also known as vitamin B9, is a water-soluble vitamin with the chemical name pteroylglutamic acid.
• Structure: It consists of three parts:
• Pteridine ring (pterin)
• Para-aminobenzoic acid (PABA)
• Glutamic acid (usually one, but can be a polyglutamate chain in natural forms)
• Formula: C19H19N7O6
Folic acid is the synthetic, oxidized form found in supplements and fortified foods; in nature, it occurs as reduced
polyglutamates (folates).
Sources
 • Rich sources: Liver, yeast, kidney, green leafy vegetables (spinach, cauliflower), beans, wheat, meat, fish, eggs.
• Fair sources: Milk, fruits.
• Bacterial synthesis: Intestinal bacteria also synthesize folate, contributing to the body's supply1.
Daily Requirement
• Adults: 400–500 µg/day
• Infants: 50 µg/day
• Children: 100–300 µg/day
• Pregnancy: 800 µg/day
• Lactation: 600 µg/day
Requirement increases during pregnancy and lactation due to increased cell division and growth1.
Coenzyme Forms
• The metabolically active form of folic acid is tetrahydrofolate (THF), also called tetrahydrofolic acid.
• In the body, folic acid is reduced to THF, which can carry and transfer single-carbon units (methyl, methylene, formyl,
formimino groups).
• Pteroylpolyglutamates are the natural coenzyme forms in tissues15.
Biological Role
• One-Carbon Metabolism: THF derivatives function as coenzymes in the transfer of single-carbon units in various
oxidation states. These are essential for:
• Synthesis of purines and pyrimidines: Needed for DNA and RNA synthesis.
 • Amino acid metabolism: Involved in the interconversion of amino acids (e.g., serine to glycine, methionine
synthesis from homocysteine).
• Methylation reactions: THF is required for the regeneration of methionine from homocysteine (with vitamin B12).
• Cell Division and Growth: Folate is critical for rapidly dividing cells (bone marrow, fetal tissues, intestinal mucosa).
• Activation of Vitamin B12: Methyl-THF is involved in converting B12 to its active coenzyme form (methylcobalamin)1.
Deficiency
• Symptoms: Glossitis, diarrhea, depression, confusion, anemia (megaloblastic or macrocytic), fatigue, mouth sores, poor
growth, and swollen tongue.
• Fetal neural tube defects: Deficiency during pregnancy can cause neural tube and brain defects in the fetus.
• Megaloblastic anemia: Due to impaired DNA synthesis, leading to large, immature red blood cells.
• Causes: Poor dietary intake, malabsorption (e.g., Crohn’s disease, celiac disease), increased requirement (pregnancy,
growth), chronic alcoholism, certain drugs (antifolates, anticonvulsants), and some genetic defects1.
Antagonists (Antifolates)
• Methotrexate, aminopterin, trimethoprim, pyrimethamine: Drugs that inhibit dihydrofolate reductase, blocking THF
formation and thus DNA synthesis. Used in cancer therapy, antibacterial, and antimalarial treatments1.

47. Vitamin B12. Chemical structure. Sources, need, coenzyme forms. Biological role.
Ans. Chemical Structure
Vitamin B12, also known as cobalamin, is a complex, water-soluble vitamin characterized by:
• A corrin ring structure similar to porphyrin but with a central cobalt ion (Co).
 • The cobalt ion is coordinated to four nitrogen atoms of the corrin ring and can form additional bonds with various
groups (e.g., cyano-, methyl-, adenosyl-, or hydroxyl- groups), resulting in different forms such as cyanocobalamin,
methylcobalamin, adenosylcobalamin, and hydroxocobalamin.
• The most common supplemental form is cyanocobalamin; the main biologically active coenzyme forms
are methylcobalamin and 5'-deoxyadenosylcobalamin2.
Sources
• Synthesized only by microorganisms (bacteria and archaea); not produced by plants or animals.
• Animal-derived foods:
• Liver, kidney (richest sources)
• Meat, fish, shellfish
• Eggs, milk, cheese
• Plant foods do not naturally contain B12 unless contaminated or fortified.
• Strict vegetarians (vegans) are at risk of deficiency unless they consume fortified foods or supplements23.
Daily Requirement
• Adults: 3 µg/day
• Infants: 0.3 µg/day
• Children: 1–2 µg/day
• Pregnancy and lactation: Increased requirement (~4 µg/day)2.
Coenzyme Forms
The metabolically active forms of vitamin B12 are:
 • Methylcobalamin:
• Coenzyme for methionine synthase, required for the remethylation of homocysteine to methionine.
• 5'-Deoxyadenosylcobalamin:
• Coenzyme for methylmalonyl-CoA mutase, which converts methylmalonyl-CoA to succinyl-CoA in the metabolism
of odd-chain fatty acids and certain amino acids25.
• Other forms: Hydroxocobalamin and cyanocobalamin (supplemental forms), aquacobalamin.
Biological Role
• DNA Synthesis and Cell Division:
• Vitamin B12 is essential for the synthesis of DNA, particularly in rapidly dividing cells such as those in bone
marrow. It is required for the conversion of ribonucleotides to deoxyribonucleotides and for thymidine synthesis2.
• Methionine and Homocysteine Metabolism:
• As a coenzyme for methionine synthase, B12 is needed for the remethylation of homocysteine to methionine,
which is also critical for the regeneration of tetrahydrofolate (THF) from methyl-THF (the "folate trap")2.
• Fatty Acid and Amino Acid Metabolism:
• 5'-Deoxyadenosylcobalamin is required for the conversion of methylmalonyl-CoA to succinyl-CoA, a step in the
catabolism of odd-chain fatty acids and certain amino acids.
• Nervous System Function:
• B12 is necessary for the maintenance of myelin sheaths around nerves, and its deficiency can lead to neurological
symptoms, including peripheral neuropathy and subacute combined degeneration of the spinal cord2.
• Red Blood Cell Formation:
 • B12 is essential for normal erythropoiesis. Deficiency leads to megaloblastic anemia, characterized by large,
immature red blood cells23.
Deficiency
• Causes:
• Poor dietary intake (especially in vegans), malabsorption (e.g., pernicious anemia due to lack of intrinsic factor,
gastric or ileal disease, post-gastric surgery), and certain genetic disorders.
• Symptoms:
• Megaloblastic (macrocytic) anemia, glossitis, stomatitis, mucosal atrophy, neurological disturbances (paresthesia,
ataxia, memory loss, subacute combined degeneration of the spinal cord), and psychiatric symptoms.
• Special Note:
• B12 deficiency can mimic folate deficiency (megaloblastic anemia), but only B12 deficiency causes neurological
symptoms23.

48. Vitamin C, chemical structure. Daily requirement, biological role, distribution in nature.
Ans. Chemical Structure
Vitamin C, also known as ascorbic acid, is a water-soluble vitamin with the chemical formula C6H8O6.
• Structurally, it is a lactone derived from glucose, containing an enediol group in the lactone ring, which is responsible for
its strong reducing (antioxidant) properties.
• The main biologically active forms are L-ascorbic acid and dehydroascorbic acid (the oxidized form, which can be
reduced back to ascorbic acid in the body)2.
Daily Requirement
 • Infants: 30 mg per day
• Adults: 75 mg per day
• Adolescents: 80 mg per day
• Pregnant women: 100 mg per day
• Lactating women: 150 mg per day
The requirement increases during infections, pregnancy, lactation, and periods of rapid growth2.
Biological Role
Vitamin C plays multiple essential roles in the body:
• Antioxidant Function:
It acts as a powerful antioxidant, protecting cells from damage by free radicals and reactive oxygen species12.
• Collagen Synthesis:
Vitamin C is a cofactor for prolyl and lysyl hydroxylases, enzymes required for hydroxylation of proline and lysine
residues during collagen synthesis. This is critical for the stability and function of connective tissue, cartilage, bone, and
teeth24.
• Wound Healing:
By supporting collagen formation, vitamin C is essential for normal wound healing and tissue repair2.
• Iron Absorption:
It enhances the absorption of non-heme iron from plant foods by reducing ferric iron (Fe³⁺) to the more absorbable
ferrous form (Fe²⁺)2.
• Synthesis of Other Molecules:
• Involved in the biosynthesis of catecholamines (e.g., norepinephrine), carnitine, and certain peptide hormones.
 • Participates in tryptophan and tyrosine metabolism2.
• Immune Function:
Supports immune defense by stimulating the activity of leukocytes and contributing to the integrity of epithelial
barriers2.
• Antioxidant Defense System:
Works synergistically with other antioxidants (e.g., vitamin E) to protect against oxidative stress1.
Distribution in Nature
Vitamin C is widely distributed in the plant kingdom and is abundant in many fruits and vegetables:
• Richest sources:
• Citrus fruits (oranges, lemons, limes)
• Amla (Indian gooseberry, one of the richest natural sources)
• Papaya, pineapple, strawberries, kiwi
• Guava, blackcurrant
• Vegetables:
• Leafy greens (spinach, cabbage)
• Broccoli, cauliflower, green peas, beans, tomatoes, potatoes
• Other:
• Germinating seeds
Note: Most animals can synthesize ascorbic acid, but humans, other primates, guinea pigs, and some bats cannot and must
obtain it from the diet2.
Vitamin C is sensitive to heat, light, and oxygen, so significant losses can occur during cooking, processing, and storage2.
Deficiency
Deficiency of vitamin C leads to scurvy, characterized by:
• Fragile capillaries and tendency to hemorrhage (petechiae, subcutaneous and internal bleeding)
• Delayed wound healing
• Swollen, bleeding gums, and loose teeth
• Poor bone and dentin formation in children
• Anemia (due to impaired iron absorption)
• General fatigue and weakness

49. Vitamin B5. Chemical structure. Sources, need. Biological role.

Ans. Chemical Structure
Vitamin B5, known as pantothenic acid, is a water-soluble vitamin.
• Structure: Pantothenic acid consists of pantoic acid and β-alanine linked by a peptide bond.
• Active form: The primary metabolically active form is coenzyme A (CoA), which is synthesized from pantothenic acid in
the body3.
Sources
Pantothenic acid is widely distributed in both plant and animal foods, hence the name (from the Greek "pantothen," meaning
"from everywhere").
• Rich sources: Liver, kidney, egg yolk, cereals, yeast, legumes3.
 • Good sources: Meats, fish, poultry, milk, whole grains, and many vegetables.
• Other: Also synthesized by intestinal bacteria, contributing to the body’s supply34.
Daily Requirement
• Adults: 5–12 mg per day (for a 2500 kcal diet)
• Infants: 1–2 mg per day
• Children: 4–5 mg per day3
Deficiency is extremely rare due to its wide distribution in foods and bacterial synthesis in the gut.
Biological Role
Pantothenic acid is essential for the synthesis and function of coenzyme A (CoA), a central molecule in metabolism. Its main
biological functions include:
• Formation of Coenzyme A:
CoA is vital for the metabolism of carbohydrates, fats, and proteins. It is required for:
• Acetyl-CoA formation: Central to the citric acid (Krebs) cycle, fatty acid synthesis and oxidation, and ketone body
metabolism.
• Succinyl-CoA formation: Important for heme synthesis and the degradation of certain amino acids and ketone
bodies.
• Lipid Metabolism:
CoA is required for β-oxidation (breakdown) and biosynthesis of fatty acids, as well as for the synthesis of cholesterol
and steroid hormones.
• Synthesis of Adrenocortical Hormones:
CoA is essential for the formation of steroid hormones from cholesterol in the adrenal cortex.
 • Activation of Amino Acids:
Some amino acids (e.g., valine, leucine) are activated to their CoA derivatives for further metabolism.
• Other Functions:
CoA is involved in acetylation reactions, which are important in the regulation of protein function and gene expression.
Deficiency:
Pantothenic acid deficiency is extremely rare in humans and has been observed only in experimental settings. Symptoms
include gastrointestinal disturbances, fatigue, sleep disturbances, neurological symptoms (staggering gait, mental changes),
and hypoglycemia3.

50. Hormones. Signs characteristic of hormones. The role of hormones in the regulation of metabolism and organ functions.
Ans. What Are Hormones?
Hormones are chemical messengers produced by specialized cells, typically in endocrine glands, that are secreted into the
bloodstream or extracellular fluid. They travel to distant target organs or tissues to regulate their structure and function,
coordinating a wide range of physiological processes across the body1.
Signs Characteristic of Hormones
• Produced in One Site, Act at Another: Hormones are synthesized in specific glands or tissues and act on target cells
located elsewhere in the body, often at a considerable distance from their site of origin1.
• Secreted in Small Quantities: They are highly potent and required only in minute amounts (often nanograms or
picograms per milliliter of blood)1.
• Transported via Blood or Extracellular Fluid: Most hormones enter the bloodstream for distribution, though some act
locally (paracrine or autocrine action)1.
• Specificity: Hormones affect only target cells that possess the appropriate receptors, ensuring precise regulation1.
 • Not Consumed in Reactions: Like enzymes, hormones are not consumed during their action and can be reused1.
• Diverse Chemical Nature: Hormones can be proteins/peptides, amino acid derivatives, steroids, or fatty acid derivatives
(e.g., eicosanoids)1.
• Regulated Secretion: Hormone release is tightly controlled by feedback mechanisms (often negative feedback), neural
stimuli, or other hormones1.
Classification of Hormones (Summary)
• By chemical structure: Steroid hormones, amino acid derivatives, peptide/protein hormones, fatty acid derivatives1.
• By site of production: Classical endocrine glands (pituitary, thyroid, adrenal, pancreas, gonads), specialized tissues
(kidney, GI tract, thymus, pineal gland)1.
• By mechanism of action: Hormones with cell surface receptors (e.g., insulin, epinephrine), hormones with intracellular
receptors (e.g., steroids, thyroid hormones)1.
Role of Hormones in Regulation of Metabolism and Organ Functions
Hormones are central to maintaining homeostasis and orchestrating the body's response to internal and external changes.
Their regulatory roles include:
1. Metabolic Regulation
• Carbohydrate Metabolism:
• Insulin lowers blood glucose by promoting glucose uptake and storage (glycogenesis), and inhibiting
gluconeogenesis and glycogenolysis25.
• Glucagon and epinephrine raise blood glucose by stimulating glycogen breakdown and gluconeogenesis25.
• Cortisol increases gluconeogenesis and reduces glucose uptake in tissues2.
• Growth hormone and thyroid hormones also modulate glucose metabolism5.
 • Lipid Metabolism:
• Insulin promotes fat storage and inhibits lipolysis26.
• Glucagon, epinephrine, and growth hormone stimulate lipolysis and mobilization of fatty acids26.
• Cortisol enhances lipolysis and fatty acid oxidation26.
• Protein Metabolism:
• Insulin stimulates protein synthesis and inhibits protein breakdown2.
• Cortisol promotes protein breakdown (catabolism) and amino acid mobilization for gluconeogenesis2.
• Growth hormone stimulates protein synthesis and cell growth1.
2. Regulation of Organ and System Functions
• Growth and Development:
• Growth hormone, thyroid hormones, sex steroids (testosterone, estrogen), and insulin-like growth
factors regulate growth, tissue development, and maturation1.
• Water and Electrolyte Balance:
• Aldosterone (adrenal cortex) controls sodium and potassium balance, affecting blood pressure and extracellular
fluid volume2.
• Antidiuretic hormone (ADH/vasopressin) regulates water reabsorption in the kidneys, maintaining plasma
osmolality1.
• Calcium and Phosphate Homeostasis:
• Parathyroid hormone (PTH), calcitonin, and vitamin D regulate blood calcium and phosphate levels, essential for
bone health and neuromuscular function1.
 • Reproductive Functions:
• Gonadotropins (FSH, LH), sex steroids, and prolactin control sexual development, gametogenesis, menstrual
cycles, pregnancy, and lactation12.
• Stress Response:
• Catecholamines (epinephrine, norepinephrine) and cortisol mediate the body's response to stress, increasing
heart rate, blood pressure, and energy availability2.
• Digestive Functions:
• Gastrointestinal hormones (gastrin, secretin, cholecystokinin) regulate digestive enzyme secretion, gastric acid
production, and gut motility2.
3. Feedback and Homeostasis
• Hormones operate via complex feedback loops (mainly negative feedback) to maintain physiological parameters within
narrow ranges, ensuring stability in the internal environment1.

51. Classification of hormones according to the place of production.

Ans. Hormones can be classified based on their site of synthesis in the body. This classification distinguishes between
hormones produced by classical endocrine glands, those produced by specialized glandular tissues, and those synthesized in
non-endocrine tissues.
1. Classical (True) Endocrine Glands
These are well-defined glands whose primary function is hormone production and secretion directly into the bloodstream.
Major classical endocrine glands include:
• Pituitary gland (anterior and posterior lobes): Growth hormone, prolactin, ACTH, TSH, FSH, LH, vasopressin, oxytocin
 • Thyroid gland: Thyroxine (T4), triiodothyronine (T3), calcitonin
• Parathyroid glands: Parathyroid hormone (PTH)
• Adrenal glands:
• Cortex: Corticosteroids (cortisol, aldosterone), androgens
• Medulla: Catecholamines (epinephrine, norepinephrine)
• Pancreas (islets of Langerhans): Insulin, glucagon, somatostatin, pancreatic polypeptide
• Ovaries: Estrogens, progesterone
• Testes: Testosterone
2. Specialized Glandular Tissues and Organs
These are tissues or organs that are not strictly endocrine glands but have specialized cells capable of producing hormones:
• Kidney: Erythropoietin (stimulates red blood cell production), renin
• Thymus: Thymosins (regulate immune function)
• Pineal gland: Melatonin (regulates circadian rhythms)
• Heart: Atrial natriuretic peptide (ANP, regulates blood pressure and volume)
• Placenta (during pregnancy): Human chorionic gonadotropin (hCG), human placental lactogen, estrogens, progesterone
3. Gastrointestinal Tract (Enteroendocrine Cells)
The GI tract contains specialized endocrine cells that secrete hormones involved in digestion and metabolism:
• Gastrin: Stimulates gastric acid secretion (stomach)
• Cholecystokinin (CCK): Stimulates pancreatic enzyme secretion and gallbladder contraction (duodenum)
 • Secretin: Stimulates bicarbonate secretion from pancreas (duodenum)
• Ghrelin: Stimulates appetite (stomach)
• Gastric inhibitory peptide (GIP), motilin, somatostatin: Various regulatory roles
4. Other Non-Endocrine Tissues with Endocrine Function
Many other tissues and organs produce hormones or hormone-like factors:
• Adipose tissue: Leptin (regulates appetite and energy balance), adiponectin
• Liver: Insulin-like growth factors (IGFs), angiotensinogen
• Skin: Vitamin D (cholecalciferol, after UV exposure)
• Endothelium: Endothelins, nitric oxide (vasoactive substances)

52. Classification of hormones by chemical structure and biological function.

Ans. Classification of Hormones by Chemical Structure and Biological Function
Classification by Chemical Structure
Hormones are classified into four main groups based on their chemical structure1:
1. Steroid Hormones
• Structure: Derived from cholesterol, containing a characteristic four-ring steroid nucleus.
• Examples: Adrenocorticosteroids (cortisol, aldosterone), sex hormones (androgens-testosterone, estrogens,
progesterone).
• Properties: Lipid-soluble, can cross cell membranes, act on intracellular receptors.
2. Amino Acid Derivatives
 • Structure: Derived from single amino acids, mainly tyrosine or tryptophan.
• Examples: Catecholamines (epinephrine, norepinephrine), thyroid hormones (thyroxine/T4, triiodothyronine/T3),
melatonin.
• Properties: Can be water-soluble (catecholamines) or lipid-soluble (thyroid hormones).
3. Peptide and Protein Hormones
• Structure: Chains of amino acids, ranging from small peptides to large proteins.
• Examples: Insulin, glucagon, parathyroid hormone (PTH), calcitonin, pituitary hormones (growth hormone, ACTH,
TSH, FSH, LH, prolactin), oxytocin, vasopressin.
• Properties: Water-soluble, act via cell surface receptors.
4. Fatty Acid Derivatives (Eicosanoids)
• Structure: Derived from polyunsaturated fatty acids, mainly arachidonic acid.
• Examples: Prostaglandins, prostacyclins, leukotrienes, thromboxanes.
• Properties: Local action (paracrine/autocrine), rapid metabolism, act via cell surface receptors.
Classification by Biological Function
Hormones can also be classified according to their principal biological effects or the systems they regulate1:
1. Metabolic Hormones
• Function: Regulate metabolism of carbohydrates, fats, and proteins.
• Examples: Insulin, glucagon, cortisol, growth hormone, thyroid hormones.
2. Growth and Development Hormones
• Function: Control growth, cell division, and differentiation.
 • Examples: Growth hormone, thyroid hormones, sex steroids (testosterone, estrogen), insulin-like growth factors
(IGFs).
3. Reproductive Hormones
• Function: Regulate sexual development, gametogenesis, menstrual cycle, pregnancy, and lactation.
• Examples: Estrogens, progesterone, testosterone, luteinizing hormone (LH), follicle-stimulating hormone (FSH),
prolactin, human chorionic gonadotropin (hCG).
4. Water and Electrolyte Balance Hormones
• Function: Maintain fluid and electrolyte homeostasis.
• Examples: Antidiuretic hormone (ADH/vasopressin), aldosterone, atrial natriuretic peptide (ANP), parathyroid
hormone (PTH), calcitonin.
5. Stress Response Hormones
• Function: Mediate the body’s response to stress (physical, emotional).
• Examples: Cortisol, catecholamines (epinephrine, norepinephrine), glucagon.
6. Digestive Hormones
• Function: Regulate secretion of digestive enzymes, acid, and gut motility.
• Examples: Gastrin, secretin, cholecystokinin (CCK), motilin, ghrelin.
7. Other Regulatory Hormones
• Function: Control circadian rhythms, immune responses, and other specialized functions.
• Examples: Melatonin (circadian rhythm), thymosins (immune function), erythropoietin (red blood cell
production).
53. Central regulation of the endocrine system: the role of liberins, statins, tropic hormones.
Ans. The endocrine system is centrally regulated by a hierarchical axis involving the hypothalamus, pituitary gland,
and peripheral endocrine glands. This system ensures precise control over hormone secretion and coordination of
physiological processes throughout the body.
1. Liberins (Releasing Hormones)
• Definition: Liberins, also known as releasing hormones or releasing factors, are peptide hormones produced by the
hypothalamus.
• Function: They stimulate the anterior pituitary gland to secrete specific tropic hormones.
• Examples:
• Corticotropin-releasing hormone (CRH or corticoliberin): Stimulates release of adrenocorticotropic hormone
(ACTH).
• Thyrotropin-releasing hormone (TRH or thyroliberin): Stimulates release of thyroid-stimulating hormone (TSH).
• Gonadotropin-releasing hormone (GnRH or luliberin): Stimulates release of luteinizing hormone (LH) and follicle-
stimulating hormone (FSH).
• Growth hormone-releasing hormone (GHRH or somatoliberin): Stimulates release of growth hormone (GH).
• Prolactin-releasing hormone (prolactoliberin): Stimulates release of prolactin.
• Melanoliberin, folliberin: Other examples with specific pituitary targets.
Liberins are secreted into the hypophyseal portal circulation and act directly on the anterior pituitary cells1.
2. Statins (Inhibiting Hormones)
 • Definition: Statins, also called inhibiting hormones or inhibiting factors, are hypothalamic peptides that suppress the
secretion of specific pituitary hormones.
• Function: They provide negative regulation to prevent overproduction of hormones.
• Examples:
• Somatostatin: Inhibits release of growth hormone (GH) and thyroid-stimulating hormone (TSH).
• Prolactin-inhibiting hormone (dopamine/prolactostatin): Inhibits release of prolactin.
• Melanostatin: Inhibits release of melanocyte-stimulating hormone (MSH).
Statins are essential for maintaining hormonal balance and preventing hypersecretion disorders1.
3. Tropic Hormones (Tropins)
• Definition: Tropic hormones are hormones secreted by the anterior pituitary that regulate the activity of other
endocrine glands.
• Function: They act as messengers, stimulating peripheral endocrine glands to produce and release their own hormones.
• Examples:
• Thyroid-stimulating hormone (TSH): Stimulates the thyroid gland to produce T3 and T4.
• Adrenocorticotropic hormone (ACTH): Stimulates the adrenal cortex to produce glucocorticoids.
• Follicle-stimulating hormone (FSH) and Luteinizing hormone (LH): Stimulate the gonads (ovaries/testes) to
produce sex hormones and gametes.
• Growth hormone (GH): Has both direct effects and stimulates the liver to produce insulin-like growth factor 1
(IGF-1).
• Prolactin (PRL): Stimulates mammary gland growth and lactation.
Tropic hormones are under the dual control of liberins (stimulation) and statins (inhibition) from the hypothalamus, and are
themselves subject to feedback inhibition by hormones produced by their target glands1.
Central Regulatory Hierarchy
1. Hypothalamus:
• Integrates neural and hormonal signals from the CNS and internal environment.
• Secretes liberins and statins into the hypophyseal portal system.
2. Anterior Pituitary (Adenohypophysis):
• Responds to hypothalamic hormones by secreting tropic hormones.
3. Peripheral Endocrine Glands:
• Respond to tropic hormones by producing the final effector hormones (e.g., cortisol, thyroid hormones, sex
steroids).
4. Feedback Regulation:
• Effector hormones from peripheral glands exert negative feedback on both the pituitary and hypothalamus,
maintaining homeostasis and preventing excessive hormone production.

54. Mechanisms of hormone action: membrane-mediated, cytosolic.

Ans. Hormones exert their effects on target cells through two principal mechanisms, determined by their chemical nature and
the location of their receptors: membrane-mediated (cell surface receptor) and cytosolic/nuclear (intracellular
receptor) mechanisms.
1. Membrane-Mediated Mechanism (Cell Surface Receptors)
Hormones:
 • Peptide/protein hormones (e.g., insulin, glucagon, growth hormone)
• Catecholamines (e.g., epinephrine, norepinephrine)
• Eicosanoids (prostaglandins, leukotrienes)
Receptor Location:
• Plasma membrane of target cells
Mechanism:
• These hormones are water-soluble and cannot cross the lipid bilayer of the cell membrane.
• They bind to specific receptors on the cell surface.
• Hormone binding triggers a conformational change in the receptor, activating intracellular signaling cascades
through second messengers.
Key Steps:
1. Hormone binds to cell surface receptor (e.g., G-protein coupled receptor, tyrosine kinase receptor).
2. Activation of second messenger systems:
• cAMP pathway: Adenylate cyclase is activated, increasing cAMP, which activates protein kinase A (PKA) (e.g.,
glucagon, epinephrine).
• cGMP pathway: Guanylate cyclase is activated, increasing cGMP, which activates protein kinase G (PKG) (e.g.,
atrial natriuretic peptide).
• Phospholipase C pathway: Generates inositol triphosphate (IP₃) and diacylglycerol (DAG), leading to increased
intracellular Ca²⁺ and activation of protein kinase C (PKC) (e.g., vasopressin, angiotensin II).
• Protein kinase activity: Some receptors have intrinsic kinase activity (e.g., insulin receptor).
 3. Cellular response:
• Activation or inhibition of enzymes
• Changes in ion channel permeability
• Altered gene expression (indirectly, via signaling cascades)
Result:
• Rapid effects (seconds to minutes), often modifying existing proteins or enzymes1.
2. Cytosolic (Intracellular/Nuclear) Mechanism
Hormones:
• Steroid hormones (e.g., cortisol, aldosterone, estrogen, testosterone)
• Thyroid hormones (T₃, T₄)
• Vitamin D, retinoic acid
Receptor Location:
• Cytoplasm or nucleus of target cells
Mechanism:
• These hormones are lipid-soluble and can diffuse across the cell membrane.
• In the cytoplasm or nucleus, they bind to specific intracellular receptors.
• The hormone-receptor complex undergoes a conformational change and translocates to the nucleus (if not already
there).
• It binds to hormone response elements (HREs) on DNA, acting as a transcription factor.
Key Steps:
 1. Hormone enters the cell by diffusion.
2. Binds to cytosolic or nuclear receptor, forming a hormone-receptor complex.
3. Complex binds to DNA at specific sites (HREs).
4. Modulation of gene transcription:
• Increases or decreases the synthesis of specific mRNAs.
• Leads to altered protein synthesis.
Result:
• Effects are typically slower (minutes to hours), but longer-lasting, as they involve changes in gene expression and
synthesis of new proteins1.

55. Tropic hormones of the pituitary gland. Chemical structure, mechanism of action, role in the regulation of the function of
the endocrine system.
Ans. Chemical Structure
Tropic hormones of the pituitary gland are peptide or glycoprotein hormones synthesized and secreted by the anterior
pituitary (adenohypophysis). They include:
• Thyroid-Stimulating Hormone (TSH, thyrotropin): Glycoprotein
• Adrenocorticotropic Hormone (ACTH, corticotropin): Peptide
• Follicle-Stimulating Hormone (FSH): Glycoprotein
• Luteinizing Hormone (LH): Glycoprotein
All these hormones are water-soluble, composed of amino acid chains (peptides or glycoproteins with carbohydrate side
chains)1.
Mechanism of Action
Tropic hormones act primarily through cell surface receptors on their target endocrine glands. Their mechanism involves:
• Binding to specific membrane receptors on target gland cells (e.g., thyroid, adrenal cortex, gonads)1.
• Activation of second messenger systems (mainly cAMP, but also others such as IP₃/DAG for some hormones):
• TSH, FSH, LH: Bind to G protein-coupled receptors, activate adenylate cyclase, increase cAMP, which activates
protein kinase A and triggers downstream signaling for hormone synthesis and secretion.
• ACTH: Binds to melanocortin 2 receptor, activates cAMP pathway, stimulating cortisol and other corticosteroid
synthesis and release1.
• Result: Stimulate growth, hormone synthesis, and secretion in their respective target glands.
Role in Regulation of the Endocrine System
Tropic hormones are central to the hierarchical regulation of the endocrine system, forming part of the hypothalamic-
pituitary-target gland axis:
1. Hypothalamic Control:
• The hypothalamus releases releasing hormones (liberins) (e.g., TRH, CRH, GnRH) that stimulate the pituitary to
secrete tropic hormones.
• It also produces inhibiting hormones (statins) (e.g., somatostatin, dopamine) that suppress pituitary hormone
release1.
2. Pituitary Secretion of Tropic Hormones:
• The anterior pituitary releases tropic hormones in response to hypothalamic signals:
 • TSH: Stimulates the thyroid gland to produce thyroxine (T4) and triiodothyronine (T3).
• ACTH: Stimulates the adrenal cortex to produce glucocorticoids (mainly cortisol).
• FSH and LH: Stimulate the gonads (ovaries and testes) to produce sex hormones (estrogen, progesterone,
testosterone) and gametes (ova, sperm)1.
3. Target Gland Response and Feedback:
• The target endocrine glands secrete their hormones, which exert physiological effects on metabolism, growth,
stress response, and reproduction.
• Negative feedback: Hormones produced by the target glands inhibit further release of both hypothalamic and
pituitary hormones, maintaining hormonal balance and homeostasis1.

56. Hormones of the posterior lobe of the pituitary gland. Chemical structure, mechanism of action, biological role.
Ans. The posterior lobe of the pituitary gland (neurohypophysis) releases two principal peptide hormones: vasopressin (also
called antidiuretic hormone, ADH) and oxytocin. These hormones are synthesized in the hypothalamus and transported to the
posterior pituitary for storage and release.
1. Chemical Structure
• Vasopressin (Antidiuretic Hormone, ADH):
• A peptide hormone composed of 9 amino acids (nonapeptide).
• Sequence: Cys-Tyr-Phe-Gln-Asn-Cys-Pro-Arg-Gly-NH₂.
• Contains a disulfide bond between the two cysteine residues, forming a ring structure.
• Oxytocin:
• Also a nonapeptide (9 amino acids).
 • Sequence: Cys-Tyr-Ile-Gln-Asn-Cys-Pro-Leu-Gly-NH₂.
• Has a similar ring structure due to a disulfide bond between cysteines.
Both hormones are synthesized as larger precursor proteins in hypothalamic neurons, cleaved to their active forms, and
transported down axons to the posterior pituitary for release1.
2. Mechanism of Action
Both vasopressin and oxytocin act via cell surface (membrane) receptors on their target cells:
• Vasopressin (ADH):
• Binds to V2 receptors in the kidney (collecting ducts), activating the adenylate cyclase–cAMP pathway, which
increases water reabsorption by inserting aquaporin-2 channels into the membrane.
• Binds to V1 receptors on vascular smooth muscle, leading to vasoconstriction via the phospholipase C–IP₃/Ca²⁺
pathway.
• Oxytocin:
• Binds to oxytocin receptors (G-protein coupled) on the smooth muscle of the uterus and mammary glands.
• Activates the phospholipase C–IP₃/Ca²⁺ pathway, increasing intracellular calcium and causing muscle contraction.
Both act rapidly and are inactivated by proteolytic enzymes in the blood and tissues1.
3. Biological Role
Vasopressin (ADH):
• Antidiuretic effect: Increases water reabsorption in the kidneys, reducing urine volume and concentrating urine.
Essential for maintaining body water balance and plasma osmolality.
• Vasoconstrictor effect: At higher concentrations, causes constriction of arterioles, increasing blood pressure.
 • Urea retention: Increases permeability of the renal medullary collecting ducts to urea, contributing to the hypertonicity
of the medullary interstitium and aiding water reabsorption.
• Glycogenolytic effect: Can increase glycogen breakdown in the liver by raising intracellular calcium.
Oxytocin:
• Uterine contraction: Stimulates powerful rhythmic contractions of the uterus during childbirth (labor), facilitating
delivery.
• Milk ejection (galactobolic effect): Causes contraction of myoepithelial cells around mammary alveoli and ducts, leading
to milk let-down during breastfeeding.
• Behavioral effects: Involved in social bonding, maternal behavior, and sexual reproduction (noted in recent research,
though primary clinical roles are above)1.

57. Thyroid hormones. Chemical structure, synthesis of iodothyronines, the need for iodide. Influence of thyroid hormones on
metabolism and body functions. Hypo- and hyperthyroidism, consequences. Endemic goiter.
Ans. Chemical Structure
Thyroid hormones are amino acid derivatives synthesized from tyrosine and contain iodine:
• Thyroxine (T4): Contains four iodine atoms.
• Triiodothyronine (T3): Contains three iodine atoms.
• Reverse T3 (rT3): An inactive isomer of T3.
T4 and T3 are produced by the follicular cells of the thyroid gland; T3 is the more biologically active hormone, binding thyroid
receptors with about 10 times higher affinity than T41.
Synthesis of Iodothyronines and Iodide Requirement
Key steps in thyroid hormone synthesis:
1. Iodide Uptake: The thyroid actively transports iodide (I⁻) from the blood into follicular cells.
2. Oxidation and Organification: Iodide is oxidized to iodine and attached to tyrosine residues in thyroglobulin, forming
monoiodotyrosine (MIT) and diiodotyrosine (DIT).
3. Coupling: Two DITs couple to form T4; one MIT and one DIT couple to form T3.
4. Release: Thyroglobulin is endocytosed, proteolytically cleaved, and T4/T3 are released into the bloodstream.
Iodide requirement:
Adequate dietary iodide is essential for thyroid hormone synthesis; deficiency impairs hormone production and leads to gland
enlargement (goiter)1.
Transport and Mechanism of Action
• Transport: In blood, T4 and T3 are mostly bound to thyroxine-binding globulin (TBG) and other plasma proteins.
• Cellular action: Both hormones enter target cells, bind to nuclear receptors, and regulate gene transcription, leading to
increased synthesis of proteins that drive metabolic processes1.
Effects of Thyroid Hormones on Metabolism and Body Functions
Metabolic effects:
• Increase basal metabolic rate: Stimulate oxygen consumption and heat production in most tissues (calorigenic effect).
• Carbohydrate metabolism: Increase glucose absorption, glycogenolysis, gluconeogenesis, and glucose utilization, often
leading to mild hyperglycemia2.
• Lipid metabolism: Stimulate lipolysis, increase plasma free fatty acids, enhance cholesterol synthesis and clearance.
• Protein metabolism: Promote both protein synthesis and degradation; excess leads to net catabolism.
 • Vitamin metabolism: Increase requirements for B vitamins and vitamin C; necessary for conversion of carotene to
vitamin A1.
Other physiological effects:
• Growth and development: Essential for normal growth, skeletal maturation, and brain development in children.
• Cardiovascular system: Increase heart rate, cardiac output, and blood flow.
• Nervous system: Enhance alertness, reflexes, and neuronal development.
• Reproductive system: Necessary for normal reproductive function.
Hypothyroidism
Causes: Autoimmune disease (Hashimoto's), iodine deficiency, thyroidectomy, congenital defects.
Manifestations:
• Fatigue, weight gain, cold intolerance, dry skin, hair loss
• Bradycardia, constipation, depression
• In children: growth retardation, delayed puberty, intellectual disability (cretinism)
• Laboratory: Low T4/T3, high TSH (primary hypothyroidism)
Consequences: Untreated, can lead to myxedema coma (life-threatening), developmental delays in children.
Hyperthyroidism
Causes: Graves' disease (autoimmune), toxic multinodular goiter, thyroid adenoma.
Manifestations:
• Weight loss despite increased appetite, heat intolerance, sweating
• Tachycardia, palpitations, anxiety, tremor
 • Muscle weakness, diarrhea, menstrual irregularities
• Laboratory: High T4/T3, low TSH
Consequences: Can cause atrial fibrillation, osteoporosis, muscle wasting, and in severe cases, thyrotoxic crisis (thyroid storm).
Endemic Goiter
Definition:
Enlargement of the thyroid gland due to chronic iodine deficiency in the diet, common in certain geographic regions (endemic
areas).
Pathogenesis:
Low iodide intake → decreased thyroid hormone synthesis → increased TSH secretion → thyroid hyperplasia and hypertrophy
(goiter formation).
Consequences:
• Goiter (visible neck swelling)
• Hypothyroidism with possible developmental and cognitive impairment in children (endemic cretinism)
• Preventable by adequate iodization of salt and dietary iodine supplementation

58. Pancreatic hormones. Chemical structure, mechanism of action, role in metabolism. Disturbance in exchange in diabetes
mellitus.
Ans. Main Pancreatic Hormones and Their Chemical Structure
The pancreas contains clusters of endocrine cells called the islets of Langerhans, which secrete several hormones:
• Insulin:
• Produced by β-cells (60% of islet cells).
 • Structure: A protein hormone composed of two polypeptide chains (A-chain: 21 amino acids, B-chain: 30 amino
acids) linked by disulfide bonds. Stored in complex with zinc5.
• Glucagon:
• Produced by α-cells (25% of islet cells).
• Structure: A single-chain polypeptide of 29 amino acids5.
• Somatostatin:
• Produced by δ-cells (10% of islet cells).
• Structure: A peptide of 14 amino acids5.
• Pancreatic Polypeptide:
• Produced by F-cells (5% of islet cells).
• Structure: A polypeptide of 36 amino acids.
Mechanism of Action
Insulin
• Receptor: Binds to a specific cell surface receptor with intrinsic tyrosine kinase activity.
• Signal Transduction:
• Binding causes receptor autophosphorylation and activation of downstream signaling pathways (notably the
PI3K/Akt pathway).
• Induces translocation of GLUT4 glucose transporters to the cell membrane in muscle and adipose tissue,
increasing glucose uptake.
• Stimulates or inhibits various enzymes via phosphorylation/dephosphorylation and gene expression changes57.
 • Main Target Tissues: Liver, muscle, adipose tissue.
Glucagon
• Receptor: Binds to G protein-coupled receptors on liver and adipose tissue.
• Signal Transduction:
• Activates adenylate cyclase, raising intracellular cAMP.
• cAMP activates protein kinase A (PKA), which phosphorylates key metabolic enzymes57.
• Main Target Tissues: Liver (primary), adipose tissue.
Somatostatin
• Receptor: Binds to G protein-coupled somatostatin receptors.
• Action: Inhibits secretion of both insulin and glucagon (paracrine effect), as well as growth hormone and various
gastrointestinal hormones5.
Role in Metabolism
Insulin
• Carbohydrate Metabolism:
• Lowers blood glucose by promoting cellular uptake, glycogen synthesis (glycogenesis), and glycolysis.
• Inhibits gluconeogenesis and glycogenolysis57.
• Lipid Metabolism:
• Promotes lipogenesis (fat storage), inhibits lipolysis (fat breakdown), reduces free fatty acids in blood, and
suppresses ketogenesis5.
• Protein Metabolism:
 • Stimulates protein synthesis and inhibits protein breakdown.
• Mineral Metabolism:
• Promotes intracellular uptake of potassium and phosphate.
Glucagon
• Carbohydrate Metabolism:
• Increases blood glucose by stimulating glycogenolysis and gluconeogenesis in the liver57.
• Lipid Metabolism:
• Stimulates lipolysis in adipose tissue, increases free fatty acids, and promotes ketogenesis in the liver.
• Protein Metabolism:
• Reduces protein synthesis, increases amino acid availability for gluconeogenesis.
Somatostatin
• General Role: Inhibits the secretion of insulin, glucagon, and various gastrointestinal hormones, thus acting as a
regulator of nutrient absorption and hormone balance5.
Disturbances in Diabetes Mellitus
Diabetes mellitus is a group of metabolic disorders characterized by chronic hyperglycemia due to defects in insulin secretion,
insulin action, or both.
Type 1 Diabetes Mellitus (Insulin-Dependent)
• Cause: Autoimmune destruction of β-cells leads to absolute insulin deficiency.
• Metabolic Disturbances:
• Marked hyperglycemia (high blood glucose).
 • Increased gluconeogenesis and glycogenolysis (unopposed glucagon action).
• Enhanced lipolysis and ketogenesis (risk of diabetic ketoacidosis).
• Protein catabolism and muscle wasting.
• Glycosuria (glucose in urine), polyuria, polydipsia, polyphagia, weight loss7.
Type 2 Diabetes Mellitus (Non-Insulin-Dependent)
• Cause: Insulin resistance in target tissues, often with relative insulin deficiency.
• Metabolic Disturbances:
• Hyperglycemia due to impaired glucose uptake and increased hepatic glucose output.
• Dyslipidemia (increased triglycerides, low HDL, high LDL).
• Less prone to ketosis than type 1.
• Chronic complications: microvascular (retinopathy, nephropathy, neuropathy) and macrovascular (atherosclerosis,
cardiovascular disease)7.
General Consequences of Insulin Deficiency or Resistance
• Carbohydrate metabolism: Impaired glucose uptake, increased hepatic glucose production.
• Lipid metabolism: Increased lipolysis, elevated free fatty acids, increased ketone body production (risk of ketoacidosis in
type 1).
• Protein metabolism: Increased proteolysis, negative nitrogen balance, muscle wasting.
• Electrolyte imbalance: Due to osmotic diuresis from glycosuria.
59. Hormones of the adrenal medulla, sources for biosynthesis, mechanism of action, influence on metabolism and functions
of organs.
Ans. Hormones and Sources for Biosynthesis
The adrenal medulla produces two major hormones:
• Epinephrine (adrenaline)
• Norepinephrine (noradrenaline)
Both are catecholamines, derived from the amino acid tyrosine. The biosynthetic pathway is:
1. Tyrosine → DOPA (by tyrosine hydroxylase)
2. DOPA → Dopamine (by DOPA decarboxylase)
3. Dopamine → Norepinephrine (by dopamine β-hydroxylase)
4. Norepinephrine → Epinephrine (by phenylethanolamine N-methyltransferase, PNMT)
The adrenal medulla is essentially a modified sympathetic ganglion, and its chromaffin cells release catecholamines directly
into the bloodstream in response to sympathetic stimulation (e.g., stress, hypoglycemia, exercise)2.
Mechanism of Action
Catecholamines act via cell surface adrenergic receptors (G protein-coupled receptors) on target cells:
• α-adrenergic receptors (α₁, α₂)
• β-adrenergic receptors (β₁, β₂, β₃)
Second messenger systems:
• cAMP pathway: β-receptors activate adenylate cyclase, increasing cAMP and activating protein kinase A, which
phosphorylates various enzymes.
• Phospholipase C pathway: α₁-receptors activate phospholipase C, producing IP₃ and DAG, leading to increased
intracellular Ca²⁺ and protein kinase C activation1.
Influence on Metabolism
Catecholamines have profound effects on fuel metabolism, rapidly mobilizing energy stores to meet acute stress or “fight-or-
flight” demands:
Carbohydrate Metabolism:
• Stimulate glycogenolysis (breakdown of glycogen) in liver and muscle, raising blood glucose.
• Stimulate gluconeogenesis in the liver.
• Inhibit insulin secretion (via α₂-receptors), reducing glucose uptake by tissues except brain and muscle during stress.
Lipid Metabolism:
• Stimulate lipolysis in adipose tissue (via β₃-receptors), increasing free fatty acids in plasma for energy use.
• Promote ketogenesis in the liver during prolonged stress or fasting.
Protein Metabolism:
• Minor direct effects, but may promote protein breakdown in muscle indirectly by increasing energy demand.
Effects on Organ Functions
Cardiovascular System:
• Increase heart rate (positive chronotropy) and force of contraction (positive inotropy) via β₁-receptors.
• Constrict blood vessels in skin, mucosa, and viscera (α₁-receptors), raising blood pressure.
• Dilate blood vessels in skeletal muscle (β₂-receptors), improving blood flow to muscles.
Respiratory System:
• Bronchodilation (β₂-receptors), improving airflow.
Ocular Effects:
 • Pupil dilation (mydriasis) via α₁-receptors.
Gastrointestinal Tract:
• Decrease motility and secretion (α and β receptors), diverting blood flow away from the gut.
Other Effects:
• Increase metabolic rate and heat production (calorigenic effect).
• Stimulate sweating (cholinergic sympathetic fibers).
• Prepare the body for acute physical activity (“fight-or-flight” response)2.

60. Hormones of the adrenal cortex, structure, mechanism of action, role in metabolism.
Ans. Chemical Structure
The adrenal cortex produces three main classes of steroid hormones, all derived from cholesterol:
• Glucocorticoids (primarily cortisol in humans)
• Mineralocorticoids (primarily aldosterone)
• Androgens (mainly dehydroepiandrosterone, DHEA, and androstenedione)
All adrenal cortical hormones share the cyclopentanoperhydrophenanthrene (steroid) nucleus characteristic of steroids, with
modifications that determine their specific activity25.
Mechanism of Action
Adrenal cortex hormones are lipid-soluble steroids and act via intracellular (cytosolic/nuclear) receptors:
• They diffuse through the plasma membrane of target cells.
• In the cytoplasm or nucleus, they bind specific steroid hormone receptors.
 • The hormone-receptor complex translocates to the nucleus, binds to hormone response elements (HREs) on DNA, and
modulates gene transcription.
• This leads to increased or decreased synthesis of specific proteins, producing the hormone’s physiological effects12.
Role in Metabolism
Glucocorticoids (Cortisol)
• Carbohydrate metabolism:
• Stimulate gluconeogenesis (formation of glucose from non-carbohydrate sources) in the liver.
• Decrease glucose uptake and utilization in peripheral tissues (anti-insulin effect), leading to increased blood
glucose.
• Protein metabolism:
• Increase protein catabolism in muscle and other tissues, releasing amino acids for gluconeogenesis.
• Enhance protein synthesis in the liver.
• Lipid metabolism:
• Stimulate lipolysis in adipose tissue, increasing free fatty acids and glycerol in plasma.
• Promote fat redistribution (e.g., central obesity in Cushing’s syndrome).
• Permissive effects:
• Enhance the effects of catecholamines and glucagon on metabolism.
• Anti-inflammatory and immunosuppressive effects:
• Suppress immune responses and inflammation at pharmacological doses25.
Mineralocorticoids (Aldosterone)
 • Electrolyte and water balance:
• Acts mainly on the distal tubules and collecting ducts of the kidney.
• Increases reabsorption of sodium and water, and promotes excretion of potassium and hydrogen ions.
• Regulates blood pressure and extracellular fluid volume.
• Regulation:
• Controlled by the renin-angiotensin-aldosterone system (RAAS), plasma potassium, and (to a lesser extent)
ACTH25.
Androgens (Adrenal Sex Hormones)
• Protein metabolism and development:
• Contribute to the development of secondary sexual characteristics, especially in females (pubic and axillary hair).
• Minor anabolic effects on protein metabolism.
• In males, testicular androgens are much more important2.

61. Sex hormones. Androgens, estrogens, gestagens, biosynthesis precursors, representatives, influence on metabolism.
Ans. Biosynthesis Precursors
All sex hormones are steroid hormones synthesized from the common precursor cholesterol. The biosynthetic pathway
proceeds via pregnenolone and progesterone, which are further converted into androgens, estrogens, and gestagens
(progestogens)41.
• Cholesterol → Pregnenolone → Progesterone
• From progesterone, pathways diverge:
 • Androgens (e.g., testosterone, androstenedione, DHEA)
• Estrogens (e.g., estradiol, estrone, estriol) via aromatization of androgens
• Gestagens (e.g., progesterone)
Androgens
Main representatives:
• Testosterone (primary male sex hormone)
• Androstenedione
• Dehydroepiandrosterone (DHEA)
• Androsterone
Sites of synthesis:
• Mainly in the Leydig cells of testes in males
• Also in ovaries (females) and adrenal cortex (both sexes)1
Functions and metabolic effects:
• Development of male reproductive organs and secondary sexual characteristics (muscle mass, deep voice, body hair)
• Anabolic effects: Stimulate protein synthesis, increase muscle and bone mass
• Promote erythropoiesis (red blood cell production)
• Stimulate libido and sexual behavior
• Regulate fat distribution (favoring visceral fat)
• Influence carbohydrate and lipid metabolism: Increase basal metabolic rate, promote glycogen synthesis, and can have
insulin-sensitizing effects14
Estrogens
Main representatives:
• Estradiol (E2) (most potent and prevalent in reproductive-age women)
• Estrone (E1)
• Estriol (E3)
Sites of synthesis:
• Ovarian follicles (granulosa cells) in females
• Also produced in small amounts by the adrenal cortex, testes, and via peripheral aromatization of androgens in adipose
tissue1
Functions and metabolic effects:
• Development of female reproductive organs and secondary sexual characteristics (breast development, fat distribution)
• Regulation of menstrual cycle (proliferative phase)
• Promote bone growth and closure of epiphyses
• Increase HDL and decrease LDL cholesterol (cardioprotective effect)
• Enhance sodium and water retention
• Stimulate protein synthesis and influence carbohydrate metabolism (increase insulin sensitivity)
• Support pregnancy and fetal development14
Gestagens (Progestogens)
Main representative:
• Progesterone
Sites of synthesis:
• Corpus luteum of the ovary (main source in non-pregnant women)
• Placenta (during pregnancy)
• Adrenal cortex (minor amounts in both sexes)
• Testes (small amounts in males)1
Functions and metabolic effects:
• Prepares and maintains endometrium for implantation and pregnancy
• Inhibits uterine contractions during pregnancy
• Regulates menstrual cycle (secretory phase)
• Supports early pregnancy
• Stimulates development of mammary glands
• Promotes fat deposition and increases basal body temperature
• Modulates immune response during pregnancy
• Influences carbohydrate and lipid metabolism: Promotes glycogen storage, increases insulin resistance, and can
increase lipolysis

62. Local and cellular hormones. Chemical nature, biological role. Biologically active peptides of the digestive tract.
Ans. Local and Cellular Hormones: Overview
Local and cellular hormones are signaling molecules that act near their site of synthesis, rather than being distributed widely
through the bloodstream like classical endocrine hormones. Their action can be classified as:
 • Paracrine: Affecting neighboring cells.
• Autocrine: Affecting the same cell that secretes them.
Chemical Nature
Local hormones include a variety of chemical classes:
• Peptides and proteins: e.g., somatostatin, gastrointestinal peptides.
• Amino acid derivatives: e.g., histamine (from histidine), serotonin (from tryptophan).
• Fatty acid derivatives: e.g., prostaglandins, leukotrienes, thromboxanes (collectively called eicosanoids).
• Gases: e.g., nitric oxide (NO).
These substances are generally not stored in large quantities but are synthesized and released quickly in response to specific
stimuli12.
Biological Role of Local and Cellular Hormones
Local and cellular hormones play crucial roles in:
• Regulation of local blood flow and vascular tone (e.g., nitric oxide, prostaglandins).
• Modulation of inflammation and immune response (e.g., histamine, prostaglandins, leukotrienes).
• Regulation of smooth muscle contraction and relaxation (e.g., prostaglandins, serotonin).
• Local control of secretion and absorption in organs (e.g., somatostatin, gastrointestinal peptides).
• Pain perception and neurotransmission (e.g., substance P, endorphins).
Their effects are typically short-lived and highly localized, allowing for fine-tuned regulation of tissue and organ function
without systemic hormonal changes126.
Biologically Active Peptides of the Digestive Tract
The digestive tract produces numerous biologically active peptides (gastrointestinal hormones), most of which act locally
(paracrine) or can have endocrine effects. Key examples include:
• Gastrin
• Source: G cells of the stomach.
• Chemical nature: Linear peptide, synthesized as a preprohormone.
• Main actions: Stimulates gastric acid secretion and promotes growth of the gastric mucosa24.
• Cholecystokinin (CCK)
• Source: I cells of the duodenum and jejunum.
• Chemical nature: Peptide hormone.
• Main actions: Stimulates secretion of pancreatic enzymes, contraction of the gallbladder, and slows gastric
emptying24.
• Secretin
• Source: S cells of the duodenum.
• Chemical nature: Peptide hormone.
• Main actions: Stimulates secretion of bicarbonate-rich pancreatic juice and bile, inhibits gastric acid secretion24.
• Gastric Inhibitory Peptide (GIP) / Glucose-dependent Insulinotropic Peptide
• Source: K cells of the duodenum and jejunum.
• Chemical nature: Peptide hormone.
• Main actions: Inhibits gastric motility and acid secretion; enhances insulin release in response to oral glucose24.
• Vasoactive Intestinal Peptide (VIP)
 • Source: Enteric neurons.
• Chemical nature: Peptide.
• Main actions: Relaxes intestinal smooth muscle, stimulates secretion of water and electrolytes, dilates intestinal
blood vessels, inhibits gastric acid secretion4.
• Somatostatin
• Source: D cells of the stomach, pancreas, and intestine.
• Chemical nature: Peptide (14 amino acids).
• Main actions: Inhibits secretion of many other GI hormones, suppresses gastric acid and pancreatic secretion,
slows gastric emptying and intestinal motility24.
• Motilin
• Source: M cells of the small intestine.
• Main actions: Stimulates gastrointestinal motility during fasting (migrating motor complex)2.
• Ghrelin
• Source: Stomach (enteroendocrine cells).
• Main actions: Stimulates hunger, increases gastric motility, and promotes growth hormone release24.

63. Nitrogen balance. Sources and ways of using amino acids in tissues. Need. Nutritional value of proteins.
Ans. Nitrogen Balance
Nitrogen balance is the difference between nitrogen intake (mainly from dietary proteins) and nitrogen loss (mainly via urine
as urea, but also in feces, sweat, skin, and hair).
 • Positive nitrogen balance: Intake > loss. Occurs during periods of growth (childhood, pregnancy), recovery from illness,
or muscle building.
• Negative nitrogen balance: Loss > intake. Seen in starvation, severe illness, trauma, burns, or inadequate protein intake.
• Nitrogen equilibrium: Intake ≈ loss. Typical in healthy adults not undergoing growth or significant tissue remodeling2.
Maintaining nitrogen balance is essential for normal growth, tissue repair, and overall metabolic health.
Sources and Utilization of Amino Acids in Tissues
Sources of amino acids:
• Dietary proteins: Digested and absorbed as amino acids.
• Tissue protein breakdown: Constant turnover of body proteins releases amino acids.
• De novo synthesis: Non-essential amino acids can be synthesized in the body (mainly in the liver), though essential
amino acids must come from the diet4.
Ways amino acids are used in tissues:
1. Protein synthesis: Formation of tissue proteins, plasma proteins, enzymes, hormones, and other functional molecules.
2. Synthesis of specialized products:
• Hemoglobin, globin, nucleoproteins.
• Biogenic amines (e.g., neurotransmitters), creatine, choline, purines, pyrimidines.
3. Energy production: Amino acids can be oxidized for energy, especially during fasting or illness.
4. Gluconeogenesis: Some amino acids (glucogenic) are converted to glucose.
5. Ketogenesis: Some amino acids (ketogenic) are converted to ketone bodies.
6. Formation of urea: Excess nitrogen is removed via the urea cycle for excretion4.
Protein Need
Protein requirements vary by age, sex, physiological state, and activity level:
• Adults: 100–130 g/day (about 14% of total calories); about 60% should be from animal sources for optimal amino acid
balance.
• For men (students): ~113 g/day (68 g animal protein)
• For women (students): ~96 g/day (58 g animal protein)
• Children: Higher per kg body weight (e.g., infants: 2.5 g/kg/day; children 1–3 years: 32 g/day)
• Pregnancy/lactation: Increased needs (pregnancy: +20 g/day; lactation: +40 g/day)2.
Nutritional Value of Proteins
Nutritional value depends on:
• Amino acid composition: Complete proteins contain all essential amino acids in adequate amounts (e.g., animal
proteins: eggs, milk, meat).
• Digestibility: How well the protein is digested and absorbed.
• Biological value: Proportion of absorbed protein that is retained for growth or maintenance. High biological value
proteins (animal sources) are more efficiently used than incomplete proteins (plant sources).
Complete proteins:
• Contain all essential amino acids (valine, leucine, isoleucine, threonine, lysine, methionine, phenylalanine, tryptophan).
• Examples: Eggs, milk, meat, fish.
Incomplete proteins:
• Lacking one or more essential amino acids.
 • Examples: Most plant proteins (legumes, cereals, nuts, seeds, fruits)2.
Reference protein:
• A theoretical protein with 100% digestibility and ideal amino acid composition, used as a standard for comparison.

64. Digestion of proteins. Proteinases, conditions of digestion in various parts of the gastrointestinal tract, absorption of
hydrolysis products.
Ans. Overview
Protein digestion is a multistage process that occurs throughout the gastrointestinal tract, involving sequential action by
various proteinases (proteolytic enzymes). The end products-mainly amino acids, dipeptides, and tripeptides-are absorbed by
the intestinal mucosa and enter the bloodstream for use by tissues.
1. Digestion in the Stomach
Key Enzymes:
• Pepsin: Secreted as inactive pepsinogen; activated by gastric hydrochloric acid (HCl) and autocatalysis.
• Type: Endopeptidase (cleaves peptide bonds within the protein chain).
• Optimal pH: 1.6–2.5.
• Action: Hydrolyzes peptide bonds, especially those formed by aromatic amino acids (phenylalanine, tyrosine,
tryptophan), producing large polypeptides and some oligopeptides.
• Rennin (Chymosin): Important in infants for milk protein (casein) digestion.
• Gastriscin, Gelatinase: Minor roles in protein digestion.
Conditions:
 • Acidic pH (due to HCl) denatures dietary proteins, making them more accessible to enzymatic hydrolysis.
• Mucus protects the stomach lining from autodigestion.
2. Digestion in the Small Intestine
Pancreatic Phase
Key Enzymes (all secreted as inactive zymogens):
• Trypsin: Activated from trypsinogen by enterokinase (from the intestinal mucosa). Cleaves peptide bonds formed by
basic amino acids (arginine, lysine).
• Chymotrypsin: Activated from chymotrypsinogen by trypsin. Cleaves bonds formed by aromatic amino acids.
• Elastase: Activated from proelastase by trypsin. Acts on peptide bonds involving neutral aliphatic amino acids.
• Carboxypeptidases A and B: Activated from procarboxypeptidases by trypsin. Exopeptidases that remove amino acids
from the carboxyl (C-) terminus of peptides.
• Collagenase: Acts on collagen proteins.
Conditions:
• Alkaline pH (due to bicarbonate from pancreatic juice) optimizes enzyme activity.
• Sequential activation: Trypsin activates other pancreatic zymogens.
Intestinal (Brush Border) Phase
Key Enzymes:
• Aminopeptidases: Exopeptidases that remove amino acids from the amino (N-) terminus.
• Dipeptidases and Tripeptidases: Hydrolyze dipeptides and tripeptides into free amino acids.
3. Absorption of Hydrolysis Products
 • Location: Mainly in the proximal small intestine (jejunum).
• Mechanisms:
• Active transport: Most L-amino acids and small peptides are absorbed via sodium-dependent active transporters.
• Dipeptides and tripeptides: Absorbed by specific peptide transporters and hydrolyzed to amino acids within
enterocytes.
• D-amino acids: Absorbed slowly, mainly by passive diffusion.
• Fate: Amino acids enter the portal vein and are transported to the liver and other tissues for protein synthesis, energy
production, or conversion to other nitrogenous compounds.

65. Gastric juice. Volume, composition. The role of hydrochloric acid and proteinases in digestion. Pathological components of
gastric juice and their definition.
Ans. Volume and Composition of Gastric Juice
• Daily Volume: The human stomach secretes approximately 2–2.5 liters of gastric juice per day1.
• Normal Composition:
• Water (major component)
• Electrolytes: Mainly hydrochloric acid (HCl), sodium, potassium, chloride ions
• Enzymes: Pepsin (main protease), rennin (in infants), gastriscin, gelatinase
• Mucus: Protects the stomach lining
• Intrinsic factor: Essential for vitamin B12 absorption
Role of Hydrochloric Acid (HCl) in Digestion
 • Activation of Pepsinogen: HCl converts inactive pepsinogen to active pepsin, the main gastric protease1.
• Optimal pH: Creates an acidic environment (pH 1.6–2.5), which is optimal for pepsin activity1.
• Protein Denaturation: Causes swelling and denaturation of dietary proteins, making them more accessible to enzymatic
hydrolysis1.
• Antimicrobial Action: Destroys many ingested pathogens.
• Regulation: Stimulates pancreatic secretion and production of local hormones, and promotes absorption of minerals
such as iron and copper1.
Role of Proteinases in Digestion
• Pepsin: The primary gastric endopeptidase, hydrolyzes peptide bonds within proteins, especially those involving
aromatic amino acids (phenylalanine, tyrosine, tryptophan)1.
• Secreted as pepsinogen and activated by HCl or autocatalytically.
• Produces large polypeptides and oligopeptides from dietary proteins.
• Rennin (Chymosin): Important in infants for milk protein (casein) digestion.
• Gastriscin and Gelatinase: Minor roles in protein digestion.
Protection of Gastric Mucosa:
• Mucus layer: Shields the stomach lining from the corrosive effects of acid and pepsin.
• Alkaline pH of mucosa and blood: Provides additional protection against self-digestion1.
Pathological Components of Gastric Juice and Their Definition
Certain abnormal components or changes in gastric juice can indicate pathology:
• Blood: Indicates gastrointestinal bleeding (e.g., gastric ulcer, cancer).
 • Bile: Presence suggests duodenogastric reflux.
• Pus: Seen in gastric infections or perforation.
• Increased or decreased acidity:
• Achlorhydria: Absence of HCl, seen in chronic gastritis, gastric cancer, or pernicious anemia.
• Hyperchlorhydria: Excess HCl, may occur in peptic ulcer disease.
• Undigested food: Suggests delayed gastric emptying or motility disorders.
• Pathological enzymes: Abnormal levels or presence of enzymes (e.g., increased pepsinogen in serum) may indicate
mucosal damage or atrophy.
Diagnostic Evaluation:
• Gastric juice analysis (pH, enzyme activity, presence of blood, bile, or pus) is used in the diagnosis of gastric and upper
GI tract diseases.

66. Replaceable and Essential Amino Acids. Biosynthesis of nonessential amino acids.
Ans. Essential vs. Nonessential Amino Acids
Essential (Indispensable) Amino Acids:
These are amino acids that cannot be synthesized by the human body in sufficient amounts and therefore must be obtained
from the diet.
Essential amino acids for humans:
• Phenylalanine
• Valine
• Threonine
 • Tryptophan
• Methionine
• Leucine
• Isoleucine
• Lysine
• Histidine (especially essential in infants and during growth)
Nonessential (Replaceable) Amino Acids:
These are amino acids that can be synthesized by the human body from metabolic intermediates or from other amino acids,
so they do not need to be supplied directly by the diet.
Nonessential amino acids include:
• Alanine
• Asparagine
• Aspartic acid
• Glutamic acid
• Serine
• Glycine
• Proline
• Cysteine (conditionally essential, synthesized from methionine)
• Tyrosine (conditionally essential, synthesized from phenylalanine)
• Glutamine
 • Arginine (conditionally essential, especially in children)
Conditionally essential amino acids are those that may become essential under certain physiological or pathological
conditions, such as rapid growth, illness, or metabolic disorders1.
Biosynthesis of Nonessential Amino Acids
Nonessential amino acids are synthesized mainly in the liver (and to some extent in other tissues) from common metabolic
intermediates, often by simple transamination or amidation reactions.
Major pathways:
• Transamination:
• The most common mechanism for synthesizing nonessential amino acids.
• An amino group is transferred from an amino acid (usually glutamate) to a keto acid, forming a new amino acid
and a new keto acid.
• Enzymes: Transaminases (aminotransferases), requiring pyridoxal phosphate (vitamin B6) as a coenzyme4.
• Examples of biosynthesis:
• Alanine: Pyruvate + glutamate ⇄ alanine + α-ketoglutarate (via alanine aminotransferase).
• Aspartate: Oxaloacetate + glutamate ⇄ aspartate + α-ketoglutarate (via aspartate aminotransferase).
• Glutamate: α-Ketoglutarate + NH₄⁺ (via glutamate dehydrogenase or by transamination).
• Glutamine: Glutamate + NH₄⁺ + ATP → glutamine (via glutamine synthetase).
• Asparagine: Aspartate + glutamine + ATP → asparagine + glutamate + AMP + PPi (via asparagine synthetase).
• Serine: Synthesized from 3-phosphoglycerate (glycolytic intermediate).
• Glycine: From serine (via serine hydroxymethyltransferase).
 • Proline: From glutamate.
• Cysteine: From serine and methionine (methionine provides the sulfur group).
• Tyrosine: From phenylalanine (via phenylalanine hydroxylase; thus, tyrosine is essential if phenylalanine is
deficient)14.

67. The transformation of non-absorbed amino acids in the large intestine with the participation of microflora. Formed
substances and their further transformation.
Ans. Overview
Not all dietary proteins and amino acids are absorbed in the small intestine; a portion reaches the large intestine, where they
are subject to further metabolism by the intestinal microflora. This microbial breakdown is significant for both local gut health
and systemic effects due to the absorption of some microbial metabolites.
Microbial Transformation of Amino Acids in the Large Intestine
1. Decarboxylation and Deamination:
• Decarboxylation: Removal of the carboxyl group from amino acids, forming biogenic amines (e.g., histidine →
histamine, tyrosine → tyramine, lysine → cadaverine, ornithine → putrescine, arginine → agmatine)24.
• Deamination: Removal of the amino group, forming ammonia (NH₃) and corresponding keto acids24.
2. Putrefaction:
• Putrefactive bacteria in the colon act on amino acids, especially those not digested/absorbed in the small intestine,
leading to the production of various putrefactive products.
Main Substances Formed
 • Ammonia (NH₃): Produced by deamination of amino acids. Ammonia is partly absorbed into the portal circulation and
transported to the liver, where it is detoxified via the urea cycle and excreted in urine2.
• Biogenic amines: Such as histamine, tyramine, cadaverine, putrescine, agmatine. These can have local effects on gut
motility and function, and in excess, may be toxic24.
• Phenol and p-cresol: Formed from the microbial degradation of tyrosine. Tyrosine can be decarboxylated to tyramine
(then reduced to p-cresol and demethylated to phenol), or deaminated to p-hydroxyphenylpropionic acid and further
metabolized to p-cresol and phenol4.
• Indole and skatole: Produced from tryptophan metabolism. Indole and skatole are responsible for the characteristic
odor of feces. Some indole is absorbed, conjugated in the liver (to indoxyl sulfate), and excreted in urine.
• Hydrogen sulfide (H₂S): Produced from cysteine and methionine by sulfate-reducing bacteria.
• Short-chain fatty acids (SCFAs): Some amino acids are also converted to SCFAs (e.g., acetate, propionate, butyrate),
though these are mainly from carbohydrate fermentation.
Further Transformation and Fate
• Absorption and Detoxification:
• Ammonia: Absorbed ammonia is rapidly transported to the liver, converted to urea (via the urea cycle), and
excreted in urine. Excess ammonia can be neurotoxic if liver function is impaired2.
• Phenol, p-cresol, indole, skatole: Absorbed into the portal blood, taken up by the liver, conjugated (usually with
sulfate or glucuronic acid), and excreted in urine.
• Local Effects:
• Biogenic amines and other products can influence gut motility, vascular tone, and local immune responses.
• Some products (like H₂S, phenol, p-cresol) are considered potentially toxic and may contribute to gut
inflammation or systemic toxicity if not efficiently detoxified.
 • Excretion:
• Non-absorbed products and bacterial biomass are excreted in feces.

68. Decarboxylation of amino acids. The role of vitamin B6. Formation of amines: histamine, serotonin, GABA. The role of
biogenic amines in the regulation of metabolism and body functions
Ans. Decarboxylation of Amino Acids
Decarboxylation is the enzymatic removal of the carboxyl group (−COOH−COOH) from amino acids, producing biogenic
amines and releasing carbon dioxide (CO2CO2). This reaction is catalyzed by amino acid decarboxylases.
• General reaction:
Amino acid→decarboxylaseBiogenic amine+CO2Amino aciddecarboxylaseBiogenic amine+CO2
Vitamin B6 (pyridoxal phosphate, PLP) is an essential coenzyme for all amino acid decarboxylases. PLP forms a Schiff base
with the amino acid, stabilizing the carbanion intermediate and facilitating the release of CO2CO226.
Formation of Key Biogenic Amines
1. Histamine
• Precursor: Histidine
• Enzyme: Histidine decarboxylase (PLP-dependent)
• Reaction:
Histidine → Histamine + CO2CO2
• Role: Histamine acts as a local hormone and neurotransmitter, involved in allergic reactions (vasodilation, increased
vascular permeability), gastric acid secretion, and neurotransmission6.
2. Serotonin (5-hydroxytryptamine)
 • Precursor: Tryptophan
• Enzyme: Tryptophan decarboxylase (PLP-dependent)
• Reaction:
5-hydroxytryptophan → Serotonin + CO2CO2
• Role: Serotonin is a neurotransmitter involved in mood regulation, sleep, appetite, and intestinal motility. It also acts as
a vasoconstrictor and modulates pain perception67.
3. Gamma-Aminobutyric Acid (GABA)
• Precursor: Glutamic acid (glutamate)
• Enzyme: Glutamate decarboxylase (PLP-dependent)
• Reaction:
Glutamate → GABA + CO2CO2
• Role: GABA is the principal inhibitory neurotransmitter in the central nervous system, reducing neuronal excitability and
preventing overexcitation (e.g., seizures). It plays a key role in regulating muscle tone and neuronal communication67.
Other Biogenic Amines
• Dopamine, norepinephrine, epinephrine: Derived from tyrosine via DOPA and dopamine (involving decarboxylation and
further modifications).
• Tyramine: From tyrosine.
• Putrescine, cadaverine, agmatine: From ornithine, lysine, and arginine, respectively, mainly in the gut.
Biological Role of Biogenic Amines in Metabolism and Body Functions
Biogenic amines have diverse and critical roles:
• Neurotransmission:
 • GABA: Inhibitory neurotransmitter, modulates neuronal excitability, prevents convulsions7.
• Serotonin: Regulates mood, sleep, appetite, pain, and gut motility.
• Dopamine: Controls movement, motivation, reward, and hormone release.
• Local Hormone Functions:
• Histamine: Mediates inflammation, allergic responses, gastric acid secretion, and acts as a neurotransmitter.
• Serotonin: Also acts as a local hormone in the gut, regulating peristalsis and secretion.
• Vascular Effects:
• Histamine: Vasodilation, increased capillary permeability (inflammation, allergy).
• Serotonin: Vasoconstriction or vasodilation depending on the tissue.
• Regulation of Smooth Muscle and Glands:
• Histamine and serotonin: Affect smooth muscle contraction and glandular secretion.
• Metabolic Regulation:
• Biogenic amines modulate hormone release, energy metabolism, and stress responses.
Deficiency of vitamin B6 impairs the synthesis of these amines, leading to neurological symptoms such as seizures (due to
decreased GABA), depression, and cognitive disturbances26.

69. Deamination of amino acids, types, biological significance.

Ans. What is Deamination?
Deamination is the process by which the amino group (–NH₂) is removed from an amino acid molecule, resulting in the
formation of ammonia (NH₃) and a corresponding keto acid. This reaction is a fundamental step in amino acid catabolism and
nitrogen metabolism in the body1.
Types of Deamination
1. Oxidative Deamination
• Definition: The removal of an amino group from an amino acid with the simultaneous oxidation of the carbon
skeleton.
• Main Enzyme: Glutamate dehydrogenase (primarily acts on glutamate).
• Reaction:
Glutamate + NAD(P)⁺ + H₂O → α-Ketoglutarate + NH₃ + NAD(P)H + H⁺
• Location: Mainly in the liver and kidney mitochondria.
• Significance: This is the primary route for the liberation of free ammonia from amino acids, especially via
glutamate, which collects amino groups from other amino acids through transamination first1.
2. Non-Oxidative Deamination
• Definition: Deamination that does not involve oxidation.
• Enzymes: Dehydratases and desulfhydrases.
• Examples:
• Serine and threonine can be deaminated by serine/threonine dehydratase to form pyruvate or α-
ketobutyrate and ammonia.
• Cysteine can be deaminated by desulfhydrase to form pyruvate, ammonia, and hydrogen sulfide1.
3. Transdeamination
 • Definition: A two-step process where the amino group from most amino acids is first transferred to α-
ketoglutarate (by transamination), forming glutamate, which then undergoes oxidative deamination to release
ammonia.
• Significance: This is the main pathway for ammonia production from amino acids in the body1.
Biological Significance of Deamination
• Ammonia Formation and Detoxification:
• Ammonia produced by deamination is toxic and must be quickly detoxified, mainly by conversion to urea in the
liver (urea cycle), which is then excreted in urine1.
• Energy Production:
• The carbon skeletons (keto acids) left after deamination can be used for:
• Gluconeogenesis (formation of glucose)
• Ketogenesis (formation of ketone bodies)
• Complete oxidation in the citric acid cycle for ATP production1.
• Link to Protein and Carbohydrate Metabolism:
• Deamination connects amino acid metabolism to central energy pathways, allowing amino acids to serve as
energy sources, especially during fasting or intense exercise1.
• Biosynthesis:
• Some deamination products serve as precursors for the synthesis of other important biomolecules (e.g.,
neurotransmitters, nucleotides)1.
• Regulation of Nitrogen Balance:
 • Deamination is essential for maintaining nitrogen balance and for the removal of excess amino acids from the
body1.

70. Transamation of amino acids. Connection with oxidative deamination, active forms of vitamin B6 in the exchange of amino
acids. Specificity of transaminases. Significance of transamination reactions. The diagnostic value of determining the activity of
transaminases.
Ans. Transamination of Amino Acids
What is Transamination?
Transamination is a reversible biochemical reaction in which the amino group (–NH₂) from an amino acid is transferred to a
keto acid, typically α-ketoglutarate, resulting in the formation of a new amino acid and a new keto acid. This process
does not release free ammonia (NH₃) and is a central mechanism in amino acid metabolism1.
General reaction:
Amino acid1+α-keto acid2↔α-keto acid1+Amino acid2Amino acid1+α-keto acid2↔α-keto acid1+Amino acid2
Connection with Oxidative Deamination
• Transamination collects amino groups from most amino acids onto glutamate.
• Glutamate is then typically deaminated by oxidative deamination (catalyzed by glutamate dehydrogenase), releasing
ammonia for urea synthesis.
• This two-step process-transamination followed by oxidative deamination-is called transdeamination and is a major
route for the removal of amino nitrogen from the body1.
Active Forms of Vitamin B6 in Amino Acid Metabolism
• Pyridoxal phosphate (PLP) is the active coenzyme form of vitamin B6 and is essential for transamination reactions.
 • PLP acts as a temporary carrier of amino groups, forming a Schiff base with the amino acid substrate, facilitating the
transfer of the amino group to the keto acid16.
Specificity of Transaminases
• Transaminases (also called aminotransferases) are the enzymes that catalyze transamination.
• Each transaminase is specific for certain pairs of amino acids and keto acids.
• Key examples:
• Alanine aminotransferase (ALT, GPT): Transfers the amino group from alanine to α-ketoglutarate, forming
pyruvate and glutamate.
• Aspartate aminotransferase (AST, GOT): Transfers the amino group from aspartate to α-ketoglutarate, forming
oxaloacetate and glutamate.
• Most amino acids can undergo transamination, but exceptions include lysine, threonine, proline, and hydroxyproline1.
Significance of Transamination Reactions
• Amino Acid Biosynthesis: Enables the synthesis of nonessential amino acids from corresponding keto acids.
• Amino Acid Catabolism: Channels amino groups from various amino acids to glutamate, which can then be deaminated
for nitrogen excretion.
• Metabolic Integration: Links amino acid metabolism with carbohydrate metabolism by interconverting amino acids and
TCA cycle intermediates.
• Urea Synthesis: Provides the nitrogen (via glutamate) for the urea cycle.
• No Free Ammonia: Unlike deamination, transamination does not produce free ammonia, thus preventing toxic
accumulation1.
Diagnostic Value of Determining Transaminase Activity
 • ALT (GPT) and AST (GOT) are present in high concentrations in the liver, heart, and other tissues.
• Elevated blood levels of these enzymes are important clinical markers:
• ALT: More specific for liver injury (e.g., hepatitis, liver necrosis).
• AST: Elevated in both liver and heart diseases (e.g., myocardial infarction, liver damage).
• Clinical significance: Measurement of serum transaminase activity is widely used in the diagnosis and monitoring of
liver diseases and myocardial infarction1.

71. Specific pathways of amino acid catabolism. Features of the exchange of serine, glycine and methionine.
Ans. General Overview of Amino Acid Catabolism
Amino acid catabolism involves the removal of the amino group (via transamination and deamination) and the breakdown of
the carbon skeleton, which is then funneled into central metabolic pathways such as glycolysis, the citric acid cycle, or used for
gluconeogenesis, ketogenesis, or biosynthesis of other compounds46.
Each amino acid has a specific catabolic pathway, often reflecting its structure and the position of its entry into central
metabolism.
Catabolism of Serine
• Main Pathway:
Serine is primarily degraded by serine dehydratase (a PLP-dependent enzyme) to yield pyruvate and ammonia:
• Serine → Pyruvate + NH₃
• Alternative Pathways:
• Serine can be converted to glycine via serine hydroxymethyltransferase (PLP-dependent), with the transfer of a
one-carbon unit to tetrahydrofolate (THF), forming N⁵,N¹⁰-methylene-THF:
 • Serine + THF → Glycine + N⁵,N¹⁰-methylene-THF + H₂O
• Fate of Products:
• Pyruvate can enter the citric acid cycle, be used for gluconeogenesis, or be converted to acetyl-CoA.
• One-carbon units transferred to THF are used in nucleotide and amino acid biosynthesis4.
Catabolism of Glycine
• Main Pathways:
1. Conversion to Serine:
Glycine can be converted back to serine (reversible reaction) by serine hydroxymethyltransferase:
• Glycine + N⁵,N¹⁰-methylene-THF + H₂O → Serine + THF
2. Glycine Cleavage System (Glycine Synthase):
Glycine is broken down to CO₂, NH₃, and a one-carbon unit transferred to THF:
• Glycine + THF + NAD⁺ → CO₂ + NH₃ + N⁵,N¹⁰-methylene-THF + NADH + H⁺
3. Oxidation to Glyoxylate:
Glycine can be oxidized to glyoxylate (by D-amino acid oxidase), which can be further metabolized to oxalate or
enter other pathways.
• Fate of Products:
• One-carbon units (as N⁵,N¹⁰-methylene-THF) are important for purine, thymidine, and methionine synthesis.
• CO₂ and NH₃ are excreted or used in other metabolic processes43.
Catabolism of Methionine
• Main Pathway:
Methionine is an essential amino acid and is catabolized via the methionine cycle and transsulfuration pathway:
 1. Activation to S-adenosylmethionine (SAM):
Methionine is first converted to SAM, the principal methyl group donor in the body.
• Methionine + ATP → S-adenosylmethionine (SAM)
2. Methyl Group Donation:
SAM donates its methyl group to various acceptors, becoming S-adenosylhomocysteine (SAH), which is hydrolyzed
to homocysteine.
3. Fates of Homocysteine:
• Remethylation:
Homocysteine can be remethylated to methionine (requires vitamin B12 and folate).
• Transsulfuration Pathway:
Homocysteine condenses with serine (via cystathionine β-synthase, PLP-dependent) to form cystathionine,
which is then cleaved to cysteine and α-ketobutyrate.
• Cystathionine → Cysteine + α-ketobutyrate
• α-Ketobutyrate is converted to propionyl-CoA, then to succinyl-CoA (a TCA cycle intermediate;
requires vitamin B12).
• Cysteine produced can be used for protein synthesis or further catabolized to sulfate and pyruvate43.
Key Features and Interconnections
• Serine and glycine metabolism are closely linked via serine hydroxymethyltransferase and one-carbon metabolism
(folate cycle).
• Methionine metabolism is linked to serine and glycine via the transsulfuration pathway and one-carbon units.
• Vitamins required:
• Vitamin B6 (PLP): Required for all transamination, deamination, and transsulfuration reactions.
 • Folate (THF): Accepts and donates one-carbon units.
• Vitamin B12: Required for remethylation of homocysteine to methionine.
Biomedical and Clinical Significance
• Defects in glycine cleavage can cause nonketotic hyperglycinemia (glycine encephalopathy).
• Homocysteine accumulation (due to vitamin B6, B12, or folate deficiency, or genetic defects) is associated with
cardiovascular disease and homocystinuria.
• Methionine, serine, and glycine are important for methylation reactions, nucleotide synthesis, and maintenance of
redox balance.

72. Phenylalanine and tyrosine metabolism. Features of tyrosine metabolism in different tissues: transformation of tyrosine in
melanocytes, thyroid gland, adrenal glands and nervous tissue. Diseases associated with metabolic disorders of these amino
acids: phenylketonuria, alkaptonuria, albinism, Parkinson's disease.
Ans. Phenylalanine and Tyrosine Metabolism
1. Catabolism Overview
• Phenylalanine is an essential amino acid. Its main fate in humans is hydroxylation to tyrosine by the
enzyme phenylalanine hydroxylase (requires tetrahydrobiopterin as a cofactor).
• Tyrosine is then further metabolized via multiple pathways, serving as a precursor for several important biomolecules.
2. Main Pathways of Tyrosine Metabolism
• General catabolic pathway:
Tyrosine → p-hydroxyphenylpyruvate → homogentisate → maleylacetoacetate → fumarylacetoacetate → fumarate +
acetoacetate
(Fumarate enters the TCA cycle; acetoacetate is a ketone body.)
Tissue-Specific Transformations of Tyrosine
A. Melanocytes (Skin)
• Tyrosine → DOPA → Dopaquinone → Melanin
• Enzyme: Tyrosinase (copper-dependent)
• Product: Melanin (pigment responsible for skin, hair, and eye color)
B. Thyroid Gland
• Tyrosine residues in thyroglobulin are iodinated to form monoiodotyrosine (MIT) and diiodotyrosine (DIT).
• Coupling reactions form the thyroid hormones triiodothyronine (T3) and thyroxine (T4).
• Function: Regulation of basal metabolic rate and development.
C. Adrenal Medulla (and Nervous Tissue)
• Tyrosine → DOPA → Dopamine → Norepinephrine → Epinephrine
• Enzymes: Tyrosine hydroxylase, DOPA decarboxylase, dopamine β-hydroxylase, phenylethanolamine N-
methyltransferase.
• Products: Catecholamines (dopamine, norepinephrine, epinephrine)-key neurotransmitters and hormones for stress
response, cardiovascular regulation, and CNS function6.
D. Nervous Tissue
• Dopamine (from tyrosine) is a major neurotransmitter.
• Further conversion: Dopamine can be converted to norepinephrine and epinephrine in certain neurons.
• Role: Regulation of movement, mood, reward, and autonomic functions6.
Diseases Associated with Phenylalanine and Tyrosine Metabolism
1. Phenylketonuria (PKU)
• Cause: Deficiency of phenylalanine hydroxylase.
• Effect: Phenylalanine accumulates, is converted to phenylpyruvate and other toxic metabolites.
• Symptoms: Severe intellectual disability, seizures, hypopigmentation, musty odor of urine.
• Treatment: Dietary restriction of phenylalanine.
2. Alkaptonuria
• Cause: Deficiency of homogentisate oxidase (in tyrosine catabolism).
• Effect: Accumulation of homogentisic acid, which is excreted in urine and darkens upon standing; deposits in connective
tissues (ochronosis).
• Symptoms: Dark urine, arthritis, connective tissue pigmentation.
3. Albinism
• Cause: Deficiency of tyrosinase in melanocytes.
• Effect: Impaired synthesis of melanin from tyrosine.
• Symptoms: Hypopigmentation of skin, hair, and eyes; increased sensitivity to sunlight; vision problems.
4. Parkinson’s Disease
• Pathophysiology: Degeneration of dopaminergic neurons in the substantia nigra (brain).
• Biochemical link: Reduced synthesis of dopamine from tyrosine.
• Symptoms: Resting tremor, rigidity, bradykinesia, postural instability.
• Treatment: Administration of L-DOPA (precursor of dopamine) to increase dopamine synthesis in the brain6.
73. Ways of formation and neutralization of ammonia. The mechanism of its toxic action.
Ans. Ways of Formation and Neutralization of Ammonia, and the Mechanism of Its Toxic Action
Ways of Ammonia Formation
Ammonia (NH3NH3) is generated in the body primarily through the metabolism of amino acids and other nitrogenous
compounds. The main sources include:
• Deamination of Amino Acids:
• Oxidative deamination (mainly of glutamate via glutamate dehydrogenase in the liver and kidney) releases free
ammonia.
• Non-oxidative deamination of certain amino acids (e.g., serine, threonine, cysteine) also produces ammonia.
• Transdeamination:
• Most amino acids transfer their amino group to α-ketoglutarate (transamination), forming glutamate, which is
then deaminated to release ammonia.
• Intestinal Bacterial Action:
• Bacterial putrefaction of proteins and amino acids in the large intestine produces significant amounts of ammonia,
which is absorbed into the portal circulation.
• Catabolism of Other Nitrogenous Compounds:
• Degradation of purines, pyrimidines, amines, and other nitrogen-containing molecules also yields ammonia.
• Other Sources:
• Deamination of amines (e.g., by monoamine oxidase), hydrolysis of glutamine and asparagine, and nucleotide
metabolism2.
Ways of Ammonia Neutralization
Ammonia is highly toxic, especially to the central nervous system, and must be rapidly detoxified:
• Urea Cycle (Ornithine Cycle):
• The primary pathway for ammonia detoxification in the liver. Ammonia is converted to urea, which is then
excreted in urine. The overall reaction is:
2NH3+CO2+3ATP+H2O→urea+2ADP+4Pi+AMP+2H+2NH3+CO2+3ATP+H2O→urea+2ADP+4Pi+AMP+2H+
• This cycle occurs mainly in hepatocytes and is the major route for nitrogen excretion in humans2.
• Glutamine Synthesis:
• In extrahepatic tissues (especially brain and muscle), ammonia is combined with glutamate to form glutamine
(via glutamine synthetase). Glutamine is a nontoxic carrier of ammonia and can be transported to the liver or
kidneys for further processing.
• In the kidney, glutamine can be deaminated to release ammonia for acid-base balance and excretion in urine2.
• Minor Pathways:
• Formation of other non-toxic compounds (e.g., asparagine) and direct excretion of small amounts of ammonia in
urine.
Mechanism of Ammonia Toxicity
Ammonia is particularly toxic to the brain. The main mechanisms of its toxic action include:
• Disruption of Cellular pH and Osmotic Balance:
• Ammonia readily crosses the blood-brain barrier and is converted to glutamine in astrocytes. Excess glutamine
increases osmotic pressure, leading to cerebral edema and neurological symptoms2.
• Depletion of TCA Cycle Intermediates:
 • Ammonia combines with α-ketoglutarate to form glutamate, depleting α-ketoglutarate, a key intermediate in the
TCA cycle. This impairs ATP production in neurons2.
• Altered Neurotransmitter Balance:
• Increased glutamate (from ammonia detoxification) can be further decarboxylated to GABA (an inhibitory
neurotransmitter), while depletion of α-ketoglutarate and glutamate can impair excitatory neurotransmission.
• The imbalance between excitatory and inhibitory neurotransmitters contributes to neurological symptoms such as
confusion, asterixis, and coma26.
• Other Effects:
• High ammonia levels can alter membrane potential, inhibit synaptic transmission, and cause astrocyte swelling.
Clinical Manifestations:
• Symptoms of ammonia toxicity include confusion, disorientation, slurred speech, bizarre behavior, and, in severe cases,
coma (hepatic encephalopathy)2.

74. Urea biosynthesis: sequence of reactions, summary equation. Hyperammonemia.

Ans. Sequence of Reactions in Urea Biosynthesis (Urea Cycle)
The urea cycle (also known as the ornithine cycle) is the principal pathway for the detoxification of ammonia in the liver,
converting toxic ammonia to urea for excretion. The cycle involves five main enzymatic steps, occurring partly in the
mitochondria and partly in the cytosol of hepatocytes:
1. Formation of Carbamoyl Phosphate (Mitochondria)
• Enzyme: Carbamoyl phosphate synthetase I (CPS-I)
• Substrates: Ammonia (NH₃), bicarbonate (HCO₃⁻), and 2 ATP
 • Product: Carbamoyl phosphate
• Reaction:
NH₃ + CO₂ + 2 ATP → Carbamoyl phosphate + 2 ADP + Pi
2. Formation of Citrulline (Mitochondria)
• Enzyme: Ornithine transcarbamoylase (OTC)
• Substrates: Carbamoyl phosphate and ornithine
• Product: Citrulline (transported to cytosol)
• Reaction:
Carbamoyl phosphate + Ornithine → Citrulline + Pi
3. Formation of Argininosuccinate (Cytosol)
• Enzyme: Argininosuccinate synthetase
• Substrates: Citrulline and aspartate (provides the second nitrogen)
• Product: Argininosuccinate
• Reaction:
Citrulline + Aspartate + ATP → Argininosuccinate + AMP + PPi
4. Cleavage of Argininosuccinate (Cytosol)
• Enzyme: Argininosuccinase (argininosuccinate lyase)
• Product: Arginine and fumarate (fumarate enters the TCA cycle)
• Reaction:
Argininosuccinate → Arginine + Fumarate
5. Formation of Urea (Cytosol)
• Enzyme: Arginase
• Product: Urea and ornithine (ornithine returns to mitochondria to continue the cycle)
• Reaction:
Arginine + H₂O → Urea + Ornithine
Summary Equation of the Urea Cycle
The overall reaction for the urea cycle is:
2NH3+CO2+3ATP+3H2O→Urea+2ADP+4Pi+AMP+2H+2NH3+CO2+3ATP+3H2O→Urea+2ADP+4Pi+AMP+2H+
• Two nitrogen atoms in urea: one from free ammonia (from deamination), one from aspartate.
• One carbon atom from CO₂ (bicarbonate).
• Energy: 3 ATP are consumed per urea molecule synthesized5.
Hyperammonemia
Definition:
Hyperammonemia is an abnormally high concentration of ammonia in the blood. It results from defects in the urea cycle,
severe liver dysfunction, or overwhelming ammonia production.
Causes:
• Inherited enzyme deficiencies in the urea cycle (e.g., CPS-I, OTC, argininosuccinate synthetase deficiency).
• Liver failure (e.g., cirrhosis, hepatitis).
• Excessive ammonia production (e.g., gastrointestinal bleeding, high protein intake, certain infections).
Consequences and Clinical Manifestations:
 • Toxicity to the CNS: Ammonia crosses the blood-brain barrier, leading to astrocyte swelling (via glutamine
accumulation), cerebral edema, and impaired neurotransmission.
• Symptoms: Lethargy, vomiting, confusion, tremors, seizures, coma, and, if untreated, death.
• Biochemical basis: Ammonia depletes α-ketoglutarate (needed for the TCA cycle), impairs ATP production, and disrupts
neurotransmitter balance5.
Significance:
The urea cycle is essential for preventing ammonia toxicity. Hyperammonemia is a medical emergency, especially in newborns
with inherited urea cycle disorders.

75. End products of nitrogen metabolism. Quantitative determination of creatinine, urea in the blood, methods of
determination.
Ans. Main End Products
Nitrogen metabolism in humans leads to the formation and excretion of several key end products:
• Urea: The principal end product, accounting for about 90% of nitrogen excreted in urine. Daily excretion: 12–20 g urea
nitrogen (12,000–20,000 mg).
• Creatinine: Formed from creatine phosphate breakdown in muscle. Excreted in urine at about 14–26 mg/kg/day in
males and 11–20 mg/kg/day in females.
• Uric Acid: End product of purine metabolism; excreted at 250–750 mg/day.
• Ammonia (NH₄⁺): Excreted as ammonium ions, about 140–1,500 mg/day; excretion increases during acidosis.
• Other minor products: Amino acids, small peptides, and products of amino acid decarboxylation1.
Quantitative Determination of Creatinine and Urea in Blood
Creatinine Determination
• Clinical significance:
Blood creatinine is a key marker of kidney (renal) function. Elevated levels indicate impaired glomerular filtration.
• Methods:
• Jaffe Reaction:
The most common method. Creatinine reacts with alkaline picrate to form a red-orange complex, which is
measured spectrophotometrically (colorimetric method).
• Principle:
Creatinine + picric acid (in alkaline medium) → creatinine-picrate complex (orange-red color)
• Measurement:
The intensity of the color is proportional to creatinine concentration and is measured at 520 nm.
• Limitations:
The method is subject to interference from non-creatinine chromogens (e.g., proteins, glucose, ketones),
but is widely used due to its simplicity.
• Enzymatic Methods:
More specific, involving creatininase, creatinase, and sarcosine oxidase, ultimately producing a measurable
change (e.g., color or absorbance) proportional to creatinine concentration.
• Advantage:
Less interference from other substances.
Urea Determination
• Clinical significance:
Blood urea (blood urea nitrogen, BUN) is also a marker of renal function and protein catabolism. Increased levels may
indicate renal impairment, dehydration, or increased protein breakdown.
 • Methods:
• Urease Method:
The most specific and widely used. Urea is hydrolyzed by the enzyme urease to produce ammonia and carbon
dioxide.
• Principle:
Urea + H₂O (urease) → 2 NH₃ + CO₂
• The ammonia produced is then quantified, commonly by:
• Colorimetric method: Ammonia reacts with hypochlorite and phenol to form indophenol (blue
color), measured spectrophotometrically.
• Berthelot Reaction: Ammonia reacts with phenol and hypochlorite in the presence of nitroprusside
catalyst to form a blue-colored compound.
• Measurement:
The intensity of the color is proportional to urea concentration.
• Other methods:
• Diacetyl monoxime method: Urea reacts with diacetyl monoxime under acidic conditions to form a colored
complex, measured spectrophotometrically.
• Enzymatic-UV method: Ammonia is measured by the decrease in absorbance of NADH at 340 nm during
the conversion of glutamate to α-ketoglutarate.

76. Biosynthesis of heme and hemoglobin in the body. Regulation of the process.
Ans. Heme Biosynthesis
Heme is a prosthetic group essential for hemoglobin, myoglobin, cytochromes, catalase, and peroxidase. Its synthesis occurs in
almost all tissues, but is most active in the liver (for cytochromes) and bone marrow (for hemoglobin).
Sequence of Heme Biosynthesis
1. Synthesis of δ-Aminolevulinic Acid (ALA)
• Location: Mitochondria
• Enzyme: ALA synthase (requires pyridoxal phosphate, vitamin B6)
• Reaction: Glycine + Succinyl-CoA → δ-ALA + CoA + CO₂
2. Formation of Porphobilinogen
• Location: Cytosol
• Enzyme: ALA dehydratase (porphobilinogen synthase)
• Reaction: 2 δ-ALA → Porphobilinogen + 2 H₂O
3. Formation of Hydroxymethylbilane and Uroporphyrinogen III
• Enzyme: Porphobilinogen deaminase and uroporphyrinogen III synthase
• Reaction: 4 Porphobilinogen → Hydroxymethylbilane → Uroporphyrinogen III
4. Conversion to Coproporphyrinogen III
• Enzyme: Uroporphyrinogen decarboxylase
• Reaction: Uroporphyrinogen III → Coproporphyrinogen III
5. Formation of Protoporphyrinogen IX and Protoporphyrin IX
• Location: Mitochondria (from this step onward)
• Enzymes: Coproporphyrinogen oxidase, protoporphyrinogen oxidase
 • Reaction: Coproporphyrinogen III → Protoporphyrinogen IX → Protoporphyrin IX
6. Insertion of Iron
• Enzyme: Ferrochelatase
• Reaction: Protoporphyrin IX + Fe²⁺ → Heme
Summary:
8 glycine + 8 succinyl-CoA + Fe²⁺ → Heme + 8 CO₂ + 8 CoA + 7 H₂O
Key vitamins/cofactors: Pyridoxal phosphate (B6), pantothenic acid (CoA), iron, vitamin C, copper.
Hemoglobin Biosynthesis
Hemoglobin is a tetrameric protein (2α, 2β chains in adults) each containing a heme group.
Steps:
1. Globin synthesis:
• Occurs on ribosomes in erythroid precursor cells (bone marrow).
• Genes for α- and β-globin chains are transcribed and translated.
2. Heme synthesis:
• As above, heme is synthesized in mitochondria and cytosol.
3. Assembly:
• Heme is inserted into each globin chain.
• Four heme-globin subunits assemble to form functional hemoglobin.
Regulation of Heme and Hemoglobin Synthesis
• Rate-limiting enzyme: δ-ALA synthase (first step) is the main regulatory point.
 • Feedback inhibition: Heme (and hemin) inhibits δ-ALA synthase by negative feedback (represses enzyme synthesis and
activity).
• Iron availability: Required for the final step (ferrochelatase); iron deficiency impairs heme synthesis.
• Erythropoietin: Stimulates hemoglobin synthesis in response to hypoxia.
• Vitamins: Pyridoxal phosphate (B6) is essential for ALA synthase; deficiencies impair heme synthesis6.
Clinical note: Defects in enzymes of the pathway cause porphyrias (disorders with accumulation of porphyrin precursors).

77. Hemoglobin catabolism. The sequence of transformations. End products of heme metabolism. Formation of bilirubin and
bilirubin glucuronide. Properties of direct and indirect bilirubin.
Ans. Sequence of Hemoglobin Catabolism
1. Destruction of Erythrocytes
• Senescent red blood cells are phagocytosed by macrophages in the spleen, liver, and bone marrow.
• Hemoglobin is released and split into globin (protein, hydrolyzed to amino acids) and heme (prosthetic group).
2. Heme Degradation
• Heme oxygenase in macrophages oxidizes heme, opening the porphyrin ring to form biliverdin (a green pigment),
releasing iron (Fe²⁺) and carbon monoxide (CO).
• Biliverdin reductase reduces biliverdin to bilirubin (a yellow pigment).
3. Transport of Bilirubin
• Unconjugated (indirect) bilirubin is hydrophobic and transported in plasma bound to albumin to the liver.
Formation of Bilirubin Glucuronide (Conjugated/Direct Bilirubin)
 4. Hepatic Uptake and Conjugation
• In hepatocytes, unconjugated bilirubin is taken up and conjugated with two molecules of glucuronic acid by UDP-
glucuronyl transferase, forming bilirubin diglucuronide (conjugated or direct bilirubin).
5. Excretion into Bile
• Conjugated bilirubin is water-soluble and actively secreted into bile canaliculi, then into the intestine.
Intestinal Transformation and End Products
6. Intestinal Metabolism
• In the intestine, bacterial enzymes deconjugate bilirubin and reduce it to urobilinogen.
• Some urobilinogen is reabsorbed (enterohepatic circulation), some is excreted in urine (as urobilin, giving urine its
yellow color), and most is oxidized to stercobilin (giving feces its brown color).
End Products of Heme Metabolism
• Bilirubin (excreted in bile)
• Urobilinogen (intestinal, some reabsorbed, some excreted in urine as urobilin)
• Stercobilin (feces)
• Iron (recycled)
• Carbon monoxide (exhaled)
Clinical Relevance
• Jaundice results from increased bilirubin in blood, classified as pre-hepatic (unconjugated), hepatic (mixed), or post-
hepatic (conjugated).
• Van den Bergh test distinguishes direct and indirect bilirubin in serum.
 •

78. Excretion of bilirubin and other bile pigments. Diagnostic value of determination of bile pigments.
Ans. Excretion of Bilirubin and Other Bile Pigments
Excretion Pathway of Bilirubin and Bile Pigments
1. Formation and Hepatic Processing
• Bilirubin is produced from the breakdown of heme in macrophages (mainly in the spleen, liver, and bone
marrow).
• Unconjugated (indirect) bilirubin, which is hydrophobic and bound to albumin, is transported to the liver.
• In hepatocytes, it is conjugated with glucuronic acid (by UDP-glucuronyl transferase) to form bilirubin
diglucuronide (conjugated or direct bilirubin), which is water-soluble and nontoxic.
2. Biliary Excretion
 • Conjugated bilirubin is actively secreted into bile canaliculi and then passes through the bile ducts into the
intestine.
3. Intestinal Transformation
• In the intestine, bacterial enzymes deconjugate bilirubin and reduce it to urobilinogen.
• Urobilinogen has several fates:
• Most is oxidized to stercobilin, which gives feces their brown color.
• Some urobilinogen is reabsorbed into the portal circulation (enterohepatic circulation); a small fraction is
excreted by the kidneys as urobilin, giving urine its yellow color.
4. Other Bile Pigments
• Biliverdin (an intermediate in heme breakdown) is reduced to bilirubin.
• Stercobilin and urobilin are the final excretory products in feces and urine, respectively1.
Diagnostic Value of Determination of Bile Pigments
Bilirubin and its metabolites are key diagnostic markers for liver and biliary tract diseases.
Types of Jaundice and Diagnostic Findings
• Prehepatic (hemolytic) jaundice:
• Increased unconjugated (indirect) bilirubin in blood.
• Urine: No conjugated bilirubin, increased urobilinogen.
• Stool: Dark brown (increased stercobilin).
• Hepatic (hepatocellular) jaundice:
• Mixed increase of both unconjugated and conjugated bilirubin.
 • Urine: Conjugated bilirubin may be present; urobilinogen variable.
• Stool: Color may be normal or pale.
• Posthepatic (obstructive) jaundice:
• Increased conjugated (direct) bilirubin in blood (due to impaired excretion).
• Urine: Conjugated bilirubin present (dark urine), urobilinogen absent or decreased.
• Stool: Clay-colored (absence of stercobilin).
Laboratory Tests
• Van den Bergh Test:
• Differentiates direct (conjugated) and indirect (unconjugated) bilirubin in serum.
• Urine analysis:
• Presence of conjugated bilirubin (direct) in urine indicates cholestasis or hepatocellular damage.
• Increased urobilinogen in urine suggests increased bilirubin turnover or impaired hepatic uptake.
• Stool color:
• Absence of stercobilin (pale or clay-colored stool) is a hallmark of biliary obstruction.
Clinical Relevance
• Hyperbilirubinemia and altered excretion of bile pigments are central to the diagnosis and classification of jaundice.
• Monitoring bilirubin fractions helps distinguish between hemolytic, hepatic, and obstructive jaundice, and guides
further investigation and management1.
79. Blood bilirubin. Reference values. Characteristics of the qualitative composition. Quantitative determination methods.
Significance in the diagnosis of pigment metabolism disorders.
Ans. Blood Bilirubin: Reference Values, Qualitative Composition, Quantitative Determination, and Diagnostic Significance
Reference Values
• Total serum bilirubin:
8–20.5 µmol/L (micromoles per liter) is considered the normal reference range for total bilirubin in blood1.
• Fractional composition:
• Indirect (unconjugated) bilirubin: ~75% of total bilirubin
• Direct (conjugated) bilirubin: ~25% of total bilirubin1
Qualitative Composition
Bilirubin in blood exists in two main forms:
• Indirect (Unconjugated) Bilirubin:
• Not water-soluble, bound to albumin in plasma
• Does not react directly with Ehrlich’s diazo reagent (indirect reaction)
• Not excreted in urine, toxic at high levels
• Direct (Conjugated) Bilirubin:
• Water-soluble, conjugated with glucuronic acid in the liver
• Reacts directly with Ehrlich’s diazo reagent (direct reaction)
• Can be excreted in urine if elevated, nontoxic1
Quantitative Determination Methods
• Van den Bergh Reaction:
• The classic method for measuring bilirubin fractions.
 • Direct (conjugated) bilirubin reacts immediately with diazo reagent (direct positive reaction).
• Indirect (unconjugated) bilirubin reacts only after addition of alcohol (indirect positive reaction).
• Total Bilirubin:
• Measured by reacting serum with diazo reagent after adding alcohol to solubilize all bilirubin forms.
• Direct Bilirubin:
• Measured by reacting serum with diazo reagent without alcohol.
• Indirect Bilirubin:
• Calculated as:
Indirect = Total bilirubin – Direct bilirubin
• Automated Methods:
• Modern clinical laboratories use automated, spectrophotometric, or colorimetric assays based on the same
principles for rapid and accurate quantification.
Diagnostic Significance
Bilirubin measurement is crucial for diagnosing and differentiating disorders of pigment metabolism and jaundice:
• Prehepatic (hemolytic) jaundice:
• Elevated unconjugated (indirect) bilirubin due to excessive breakdown of red blood cells.
• Direct bilirubin remains normal; urine bilirubin absent; stool dark1.
• Hepatic (hepatocellular) jaundice:
• Both direct and indirect bilirubin are elevated due to impaired hepatic uptake, conjugation, or secretion.
• Mixed pattern in blood; urine may contain conjugated bilirubin1.
 • Posthepatic (obstructive) jaundice:
• Elevated direct (conjugated) bilirubin due to biliary obstruction.
• Conjugated bilirubin appears in urine; stool is pale (clay-colored)1.
Other uses:
• Monitoring liver function in hepatitis, cirrhosis, and drug-induced liver injury.
• Diagnosing hereditary disorders (e.g., Gilbert’s syndrome, Crigler-Najjar, Dubin-Johnson syndromes).

80. Pyrimidine nucleotides, representatives, biological role. Biosynthesis of pyrimidine nucleotides. Formation of dehydroorotic
acid. The role of ATP in the synthesis of pyrimidine mononucleotides. The role of free pyrimidines in metabolism (UDP, CDP).
Ans. Representatives and Biological Role
Pyrimidine nucleotides are nucleotides containing a pyrimidine base-cytosine (C), uracil (U), or thymine (T)-linked to a sugar
(ribose or deoxyribose) and phosphate group. The main representatives include:
• UMP (uridine monophosphate)
• UDP (uridine diphosphate)
• UTP (uridine triphosphate)
• CMP (cytidine monophosphate)
• CDP (cytidine diphosphate)
• CTP (cytidine triphosphate)
• dUMP, dTMP, dCMP, dCTP (deoxyribonucleotides; for DNA synthesis)
Biological roles:
 • Structural: Building blocks of RNA (C, U) and DNA (C, T)
• Energy transfer: UTP is used in carbohydrate metabolism (e.g., UDP-glucose in glycogen synthesis)
• Biosynthetic precursors: CTP for phospholipid synthesis
• Regulatory: UDP and CDP derivatives act as intermediates in various biosynthetic pathways
Biosynthesis of Pyrimidine Nucleotides
Overview
Pyrimidine biosynthesis differs from purine synthesis in that the pyrimidine ring is synthesized first and then attached to
ribose-5-phosphate.
Steps:
1. Carbamoyl Phosphate Formation
• Enzyme: Carbamoyl phosphate synthetase II (cytosolic)
• Reactants: Glutamine, CO₂, 2 ATP
• Product: Carbamoyl phosphate
2. Formation of Carbamoyl Aspartate
• Enzyme: Aspartate transcarbamoylase
• Reactants: Carbamoyl phosphate + aspartate
• Product: Carbamoyl aspartate
3. Cyclization to Dihydroorotate
• Enzyme: Dihydroorotase
• Product: Dihydroorotic acid (dihydroorotate)
 4. Oxidation to Orotic Acid
• Enzyme: Dihydroorotate dehydrogenase (mitochondrial)
• Product: Orotic acid (orotate)
5. Formation of Orotidine Monophosphate (OMP)
• Enzyme: Orotate phosphoribosyltransferase
• Reactants: Orotic acid + PRPP (5-phosphoribosyl-1-pyrophosphate)
• Product: OMP
6. Decarboxylation to Uridine Monophosphate (UMP)
• Enzyme: OMP decarboxylase
• Product: UMP
7. Phosphorylation and Amination
• UMP is phosphorylated to UDP and UTP (by kinases, using ATP).
• UTP is aminated (using glutamine and ATP) to form CTP.
Summary of ATP Role
• ATP is required at multiple steps:
• Carbamoyl phosphate synthesis (2 ATP)
• Conversion of UMP to UDP/UTP (kinases)
• Amination of UTP to CTP (uses ATP as energy source)
Formation of Dihydroorotic Acid
 • Dihydroorotic acid is formed by cyclization of carbamoyl aspartate via dihydroorotase.
• It is then oxidized to orotic acid, a key intermediate in pyrimidine biosynthesis.
Role of Free Pyrimidines in Metabolism (UDP, CDP)
• UDP (Uridine diphosphate):
• UDP-glucose: Essential for glycogen synthesis and metabolism of galactose.
• UDP-glucuronate: Used in conjugation reactions for detoxification (e.g., bilirubin conjugation).
• CDP (Cytidine diphosphate):
• CDP-choline, CDP-ethanolamine: Intermediates in phospholipid (membrane lipid) biosynthesis.
• CDP-diacylglycerol: Precursor for phosphatidylinositol and cardiolipin synthesis.
These nucleotides act as activated carriers of sugars and other groups in biosynthetic pathways, and as such are crucial for
cellular metabolism and macromolecule synthesis.

81. Decomposition of purine nitrogenous bases in tissues. Gout and hyperuricemia.

Ans. Decomposition of Purine Nitrogenous Bases in Tissues
Pathway of Purine Catabolism
Purine nucleotides (AMP and GMP) from nucleic acids are degraded in tissues through the following sequence of enzymatic
reactions:
1. Nucleotide Hydrolysis:
• AMP and GMP are hydrolyzed by purine-5'-nucleotidase to their respective
nucleosides, adenosine and guanosine.
 2. Deamination and Phosphorolysis:
• Adenosine is deaminated by adenosine deaminase to form inosine.
• Inosine and guanosine are cleaved by purine nucleoside phosphorylase to yield hypoxanthine (from inosine)
and guanine (from guanosine), plus ribose-1-phosphate.
3. Further Deamination:
• Guanine is deaminated to xanthine.
4. Oxidation:
• Hypoxanthine is oxidized by xanthine oxidase to xanthine.
• Xanthine is further oxidized by the same enzyme to uric acid.
5. Excretion:
• Uric acid is the final product of purine catabolism in humans and is excreted in urine1.
Key Points:
• Molecular oxygen is reduced during xanthine oxidase reactions, producing superoxide (O₂⁻) and hydrogen peroxide
(H₂O₂) as byproducts1.
• In most mammals, uric acid is further degraded to allantoin, but in humans and higher primates, uric acid is the end
product due to lack of uricase1.
Gout and Hyperuricemia
Hyperuricemia
• Defined as an increased concentration of uric acid in the blood.
• Reference values:
 • Men: 202–417 μmol/L
• Women: 143–339 μmol/L1
• Causes:
• Primary hyperuricemia: Often due to inherited metabolic defects leading to increased purine synthesis or
decreased uric acid excretion.
• Primary metabolic gout: Excessive conversion of glycine to uric acid.
• Primary renal gout: Impaired renal excretion of uric acid.
• Secondary hyperuricemia: Due to increased purine catabolism (e.g., leukemia, polycythemia, prolonged fasting)
or renal failure1.
Gout
• Gout is a clinical syndrome characterized by chronic elevation of uric acid (hyperuricemia) with deposition of
monosodium urate crystals in joints and tissues, leading to acute inflammatory arthritis (most commonly affecting the
big toe-podagra), tophi formation, and potential kidney stones1.
• Pathogenesis:
• When uric acid levels exceed its solubility, crystals precipitate in joints and soft tissues, triggering inflammation.
• Types:
• Primary gout: Usually genetic, due to enzyme defects in purine metabolism or renal urate handling.
• Secondary gout: Associated with other diseases (e.g., increased cell turnover in leukemia, renal failure)1.
• Von Gierke's disease: A glycogen storage disease (glucose-6-phosphatase deficiency) with increased pentose
phosphate pathway activity, leading to increased purine synthesis and hyperuricemia1.
82. Carbohydrates. Classification. Biological role. The most important representatives.
Ans. Classification of Carbohydrates
Carbohydrates are organic compounds containing carbon, hydrogen, and oxygen, typically with the general formula Cₙ(H₂O)ₙ.
They are classified based on their structure and complexity:
1. Monosaccharides (Simple Sugars)
• Cannot be hydrolyzed into simpler carbohydrates.
• Classified by:
• Number of carbon atoms: trioses (3C), tetroses (4C), pentoses (5C), hexoses (6C), etc.
• Functional group: Aldoses (contain an aldehyde group, e.g., glucose) and Ketoses (contain a ketone group, e.g.,
fructose).
• Examples: Glucose, fructose, galactose, ribose.
2. Disaccharides
• Composed of two monosaccharide units linked by a glycosidic bond.
• Examples:
• Sucrose (glucose + fructose)
• Lactose (glucose + galactose)
• Maltose (glucose + glucose)
3. Oligosaccharides
• Contain 2–10 monosaccharide units.
• Found in glycoproteins and glycolipids.
4. Polysaccharides
• Polymers with more than 10 monosaccharide units.
• Homopolysaccharides: Made of one type of monosaccharide (e.g., starch, glycogen, cellulose).
• Heteropolysaccharides: Made of different types of monosaccharides (e.g., hyaluronic acid, heparin, chondroitin sulfate).
Biological Role of Carbohydrates
• Energy Source: The primary function; glucose is the main fuel for most cells, especially the brain and red blood cells.
• Energy Storage: Stored as glycogen in animals and starch in plants for later use.
• Structural Role:
• Cellulose provides structural support in plant cell walls.
• Glycosaminoglycans (e.g., hyaluronic acid, chondroitin sulfate) are key components of connective tissue and
extracellular matrix.
• Component of Nucleic Acids: Ribose and deoxyribose are part of RNA and DNA.
• Cell Recognition and Signaling: Oligosaccharides on glycoproteins and glycolipids are crucial for cell-cell recognition,
immune response, and signaling.
• Detoxification: Some carbohydrates participate in detoxification reactions (e.g., glucuronic acid conjugation).
Most Important Representatives

Group Examples Main Functions/Locations

Glucose, fructose, galactose, Energy, nucleic acids, metabolic

Monosaccharides ribose intermediates
 Group Examples Main Functions/Locations

Disaccharides Sucrose, lactose, maltose Dietary sugars, energy source

Energy storage (starch, glycogen),

Polysaccharides Starch, glycogen, cellulose structure (cellulose)

Hyaluronic acid, heparin, Connective tissue, anticoagulation,

Heteropolysaccharides chondroitin sulfate matrix structure

Key monosaccharides:
• Glucose: Central to metabolism, main blood sugar23.
• Fructose: Found in fruits, component of sucrose.
• Galactose: Component of lactose.
• Ribose: Component of RNA; deoxyribose in DNA.
Key polysaccharides:
• Starch: Plant storage carbohydrate (amylose and amylopectin).
• Glycogen: Animal storage carbohydrate (liver and muscle).
• Cellulose: Plant structural carbohydrate, not digestible by humans.
Key heteropolysaccharides:
• Hyaluronic acid, chondroitin sulfate, heparin: Components of extracellular matrix, cartilage, and anticoagulant
function3.
83. Polysaccharides of animal tissues. Classification. Biological role. Representatives.
Ans. Classification
Polysaccharides in animal tissues are classified based on their monomer composition and function:
1. Homopolysaccharides
• Composed of only one type of monosaccharide.
• Main example in animals: Glycogen.
2. Heteropolysaccharides
• Composed of two or more different types of monosaccharides (often amino sugars and uronic acids).
• Main examples: Glycosaminoglycans (GAGs), also known as mucopolysaccharides.
Biological Role
Polysaccharides in animal tissues serve several essential functions:
• Energy Storage:
• Glycogen acts as the primary storage form of glucose in animals, especially in liver and muscle, providing a readily
mobilizable energy reserve34.
• Structural and Support Functions:
• Glycosaminoglycans (GAGs) are major components of the extracellular matrix, providing structural support,
viscosity, and elasticity to connective tissues, cartilage, skin, and synovial fluid14.
• Protection and Lubrication:
 • Mucopolysaccharides (GAGs) contribute to the viscosity of mucus, synovial fluid, and serve as lubricants in joints
and other tissues.
• Cell Communication and Adhesion:
• Some polysaccharides (as part of glycoproteins and proteoglycans) are involved in cell recognition, signaling, and
adhesion processes1.
Representatives
1. Glycogen
• Type: Homopolysaccharide (glucose units)
• Structure: Highly branched polymer of α-D-glucose with α-1,4 glycosidic bonds and α-1,6 branches every 8–12 residues.
• Location: Liver (for blood glucose regulation), skeletal muscle (for local energy supply).
• Function: Rapidly mobilizable energy storage for glucose34.
2. Glycosaminoglycans (GAGs) / Mucopolysaccharides
• Type: Heteropolysaccharides (repeating disaccharide units of amino sugars and uronic acids)
• Key Examples:
• Hyaluronic acid: Provides viscosity and lubrication in synovial fluid, vitreous humor, and connective tissues.
• Chondroitin sulfate: Major component of cartilage, provides resistance to compression.
• Dermatan sulfate: Found in skin, blood vessels, and heart valves.
• Keratan sulfate: Present in cartilage and cornea.
• Heparin: Anticoagulant found in liver and mast cells.
• Function: Structural integrity of tissues, regulation of water content, shock absorption, and anticoagulation14.
3. Glycoproteins and Proteoglycans
• Glycoproteins: Proteins with short carbohydrate chains (oligosaccharides); involved in cell recognition, immunity, and
hormone function.
• Proteoglycans: Proteins with long GAG chains; key components of the extracellular matrix, contributing to tissue
resilience and hydration1.

84. Food carbohydrates, need, digestion in the gastrointestinal tract. Absorption of hydrolysis products. Digestion disorder.
Enzymopathies.
Ans. 1. Food Carbohydrates and Daily Need
• Sources: Carbohydrates are present in foods as monosaccharides (glucose, fructose), disaccharides (sucrose, lactose,
maltose), and polysaccharides (starch, glycogen, cellulose, dietary fiber).
• Dietary Need: Carbohydrates are the primary energy source for humans, typically providing 50–60% of daily caloric
intake. The recommended daily intake for adults is about 300–400 g, with a minimum of 100–130 g/day needed to
prevent ketosis and supply the brain1.
2. Digestion of Carbohydrates in the Gastrointestinal Tract
Mouth
• Salivary α-amylase begins starch digestion, hydrolyzing internal α-1,4-glycosidic bonds to produce maltose, maltotriose,
and dextrins.
• Limited action due to short oral transit and inactivation by gastric acid1.
Stomach
• No significant carbohydrate digestion occurs due to acidic pH, which inactivates salivary amylase1.
Small Intestine
• Pancreatic α-amylase continues starch breakdown to maltose, isomaltose, and oligosaccharides.
• Brush border enzymes (disaccharidases) on intestinal epithelial cells complete digestion:
• Maltase: Maltose → 2 glucose
• Sucrase: Sucrose → glucose + fructose
• Lactase: Lactose → glucose + galactose
• Isomaltase: Isomaltose → 2 glucose
• Oligosaccharidases hydrolyze short-chain oligosaccharides1.
Large Intestine
• Undigested carbohydrates (mainly dietary fiber and resistant starches) are fermented by colonic bacteria, producing
short-chain fatty acids (SCFAs), gases, and other metabolites1.
3. Absorption of Hydrolysis Products
• Monosaccharides (glucose, galactose, fructose) are absorbed mainly in the small intestine:
• Glucose and galactose: Active transport via sodium-dependent SGLT1 transporter.
• Fructose: Facilitated diffusion via GLUT5 transporter.
• All monosaccharides exit enterocytes into the portal blood via GLUT2 transporter1.
• Pentoses (e.g., ribose) are absorbed slowly by passive diffusion.
4. Disorders of Digestion and Absorption (Malabsorption)
Lactase Deficiency (Lactose Intolerance)
• Cause: Deficiency of lactase enzyme in the brush border.
 • Symptoms: Diarrhea, flatulence, abdominal cramps, distension after dairy intake.
• Mechanism: Undigested lactose is fermented by colonic bacteria, producing gas and osmotic diarrhea1.
Sucrase Deficiency
• Cause: Inherited or acquired deficiency of sucrase-isomaltase complex.
• Symptoms: Similar to lactase deficiency but triggered by sucrose-containing foods1.
Disacchariduria
• Definition: Increased excretion of disaccharides in urine due to disaccharidase deficiency or intestinal mucosal damage
(e.g., celiac disease, sprue)1.
Monosaccharide Malabsorption
• Cause: Rare, often due to absence of specific carrier proteins for glucose/galactose.
• Symptoms: Severe diarrhea and failure to thrive in infants1.
5. Enzymopathies (Enzyme Deficiencies) in Carbohydrate Digestion
• Congenital or acquired deficiencies of digestive enzymes lead to malabsorption syndromes:
• Lactase deficiency: Most common; leads to lactose intolerance1.
• Sucrase-isomaltase deficiency: Rare; leads to sucrose and isomaltose intolerance.
• Glucose-galactose malabsorption: Due to SGLT1 transporter defect; severe, requires dietary fructose as only
carbohydrate source.
• Secondary enzymopathies may occur due to intestinal diseases (e.g., celiac disease, infections) that damage the mucosa
and reduce enzyme activity1.
85. Amylase, determination in biological fluids, diagnostic value, study of activity.
Ans. What is Amylase?
Amylase is a hydrolase enzyme (EC 3.2.1.1) that catalyzes the hydrolysis of starch and glycogen into smaller sugars such as
maltose and dextrins. It is produced mainly by the pancreas (pancreatic amylase) and salivary glands (salivary amylase).
Determination of Amylase in Biological Fluids
Amylase activity is most commonly measured in serum (blood plasma) and urine. The determination is based on its ability to
hydrolyze specific substrates.
Principles of Amylase Assay
• Enzyme assays measure either the consumption of substrate or the formation of product over time.
• Direct assay: Substrate is added to the biological fluid, and the amount of product formed (e.g., maltose, glucose) is
determined.
• Indirect assay: The enzyme is extracted and then its activity is measured.
Methods of Determination
• Colorimetric Methods:
• Starch is used as a substrate. After incubation, the decrease in starch (iodine-starch complex) or the increase in
reducing sugars (e.g., maltose, glucose) is measured by color change.
• Chromogenic Substrates:
• Artificial substrates that release a colored product upon hydrolysis by amylase.
• Spectrophotometric Methods:
• The rate of substrate breakdown or product formation is measured by changes in absorbance at a specific
wavelength.
 • Enzyme Unit Definition:
• One unit (U) of amylase activity is the amount of enzyme that catalyzes the conversion of one micromole of
substrate per minute under specified conditions1.
Sample Types
• Serum (plasma): Most common for clinical diagnostics.
• Urine: Useful in some acute and chronic conditions (e.g., pancreatitis).
Diagnostic Value
Amylase is a key diagnostic enzyme in medicine, especially for pancreatic and salivary gland disorders:
• Acute Pancreatitis:
• Marked increase in serum and urine amylase activity is a classic marker.
• Levels rise within hours of onset, peak at 12–72 hours, and return to normal in 3–5 days.
• Chronic Pancreatitis and Pancreatic Insufficiency:
• May show persistently low or normal amylase due to loss of pancreatic tissue.
• Salivary Gland Diseases:
• Elevated in conditions like mumps (parotitis).
• Other Conditions:
• Increased in perforated peptic ulcer, intestinal obstruction, renal failure, and some gynecological diseases.
Interpretation:
• High serum amylase: Suggests acute pancreatitis, salivary gland inflammation, or macroamylasemia.
• High urine amylase: Supports diagnosis of acute pancreatitis or renal impairment.
 • Low amylase: May indicate chronic pancreatic insufficiency.
Study of Amylase Activity
Factors affecting assay results:
• Temperature, pH, substrate concentration, ionic strength must be controlled for reliable results1.
• Enzyme activity is expressed in units per liter (U/L) in clinical practice.
Clinical Application:
• Routine part of biochemical panels for patients with abdominal pain or suspected pancreatic disease.
• Serial measurements can monitor disease progression or response to therapy.

86. Carbohydrate reserves of the body. Structure, biological functions of glycogen. Biosynthesis, localization and regulation of
the process. Mobilization of glycogen, sequence of reactions. The mechanism of hormonal regulation of biosynthesis and
breakdown of glycogen. Glyco- and aglycogenoses.
Ans. Structure and Biological Functions of Glycogen
Glycogen is the main carbohydrate reserve in animals, found primarily in the liver and skeletal muscles. It is a highly branched
homopolysaccharide composed of α-D-glucose units linked by α-1,4-glycosidic bonds in the linear chains and α-1,6-glycosidic
bonds at the branch points (every 8–10 glucose residues)24. This structure allows rapid mobilization and synthesis, as enzymes
can simultaneously act on many non-reducing ends.
Biological functions:
• Energy reserve: Glycogen serves as a readily mobilizable glucose store to maintain blood glucose (liver) and provide
energy for muscle contraction (muscle).
• Buffer: Helps maintain blood glucose between meals and during fasting.
 • Rapid energy source: Especially important during sudden, intense muscular activity.
Biosynthesis (Glycogenesis), Localization, and Regulation
Localization:
• Liver: ~100–120 g in adults; maintains blood glucose.
• Muscle: ~300–400 g; provides energy for muscle activity.
Biosynthesis steps:
1. Glucose → Glucose-6-phosphate
• Enzyme: Hexokinase (muscle) or glucokinase (liver).
2. Glucose-6-phosphate ↔ Glucose-1-phosphate
• Enzyme: Phosphoglucomutase.
3. Glucose-1-phosphate + UTP → UDP-glucose
• Enzyme: UDP-glucose pyrophosphorylase.
4. UDP-glucose + Glycogen (primer) → Glycogen (elongated) + UDP
• Enzyme: Glycogen synthase (forms α-1,4 bonds).
5. Branching:
• Enzyme: Branching enzyme (amylo-1,4→1,6-transglycosylase) creates α-1,6 linkages2.
Regulation:
• Insulin: Stimulates glycogen synthesis by activating glycogen synthase and inhibiting glycogen phosphorylase12.
• Glucagon (liver) and adrenaline (muscle/liver): Inhibit glycogen synthesis by activating kinases that phosphorylate and
inactivate glycogen synthase.
Mobilization of Glycogen (Glycogenolysis): Sequence of Reactions
1. Glycogen (n) + Pi → Glycogen (n-1) + Glucose-1-phosphate
• Enzyme: Glycogen phosphorylase (cleaves α-1,4 bonds).
2. Debranching enzyme:
• Removes branches by transferring short glucose chains and hydrolyzing α-1,6 bonds to release free glucose.
3. Glucose-1-phosphate ↔ Glucose-6-phosphate
• Enzyme: Phosphoglucomutase.
4. Liver/kidney only:
• Glucose-6-phosphate → Glucose (enzyme: glucose-6-phosphatase), allowing glucose release into blood2.
In muscle: Glucose-6-phosphate enters glycolysis for ATP production (muscle lacks glucose-6-phosphatase).
Hormonal Regulation of Glycogen Metabolism

Hormone Effect on Effect on Mechanism

Glycogen Glycogen
Synthesis Breakdown

Activates phosphatases
(dephosphorylation), activates synthase,
Insulin Stimulates Inhibits inhibits phosphorylase12
 Hormone Effect on Effect on Mechanism
Glycogen Glycogen
Synthesis Breakdown

Activates cAMP pathway, activates kinases,

Stimulates phosphorylates and activates
Glucagon Inhibits (liver) (liver) phosphorylase, inactivates synthase12

Inhibits Similar to glucagon; acts via β-adrenergic

Adrenaline (muscle/liver) Stimulates receptors, cAMP12

• Insulin promotes glycogen storage after meals.

• Glucagon (liver) and adrenaline (muscle/liver) promote glycogen breakdown during fasting, stress, or exercise.
Glycogen Storage Diseases (Glyco- and Aglycogenoses)
• Glycogenoses: Genetic disorders caused by enzyme deficiencies in glycogen metabolism, leading to abnormal storage or
structure of glycogen.
• Examples:
• Von Gierke’s disease (Type I): Glucose-6-phosphatase deficiency-severe hypoglycemia, hepatomegaly.
• Pompe’s disease (Type II): Lysosomal α-1,4-glucosidase deficiency-cardiomegaly, muscle weakness.
• Cori’s disease (Type III): Debranching enzyme deficiency-limit dextrin accumulation.
• McArdle’s disease (Type V): Muscle phosphorylase deficiency-exercise intolerance, muscle cramps.
• Aglycogenoses: Rare, total absence of glycogen due to lack of glycogen synthase.
87. Ways of glucose oxidation in tissues. Characteristics of anaerobic breakdown of glucose: localization in the cell, prevalence
in the body, sequence of reactions, biological role.
Ans. Ways of Glucose Oxidation in Tissues
Glucose, as the central energy substrate, is oxidized in tissues through several major metabolic pathways:
Major Pathways of Glucose Oxidation
1. Glycolysis (Embden-Meyerhof-Parnas pathway)
• Converts glucose to pyruvate (aerobically) or lactate (anaerobically).
• Occurs in the cytosol of all cells1.
2. Aerobic Oxidation
• Pyruvate from glycolysis is converted to acetyl-CoA (in mitochondria), enters the citric acid (TCA) cycle, and is fully
oxidized to CO₂ and H₂O via the electron transport chain and oxidative phosphorylation12.
3. Pentose Phosphate Pathway (Hexose Monophosphate Shunt)
• Oxidizes glucose-6-phosphate to generate NADPH and ribose-5-phosphate for biosynthetic processes, not
primarily for ATP production1.
Anaerobic Breakdown of Glucose (Anaerobic Glycolysis)
Localization in the Cell
• Cytosol: All glycolytic reactions, including anaerobic steps, occur in the cytoplasm1.
Prevalence in the Body
• Universal pathway: Occurs in all tissues.
 • Essential in tissues with low or no mitochondria: Erythrocytes (no mitochondria), renal medulla, lens and cornea of the
eye, exercising skeletal muscle, some brain regions, and cancer cells rely heavily on anaerobic glycolysis1.
• Occurs during hypoxia or intense muscular activity: When oxygen supply is insufficient for aerobic metabolism.
Sequence of Reactions (Glycolysis to Lactate)
The anaerobic breakdown of glucose involves the following steps:
1. Glucose → Glucose-6-phosphate
• Enzyme: Hexokinase (or glucokinase in liver)
• ATP consumed
2. Glucose-6-phosphate → Fructose-6-phosphate
• Enzyme: Phosphoglucose isomerase
3. Fructose-6-phosphate → Fructose-1,6-bisphosphate
• Enzyme: Phosphofructokinase-1 (PFK-1)
• ATP consumed
4. Fructose-1,6-bisphosphate → Glyceraldehyde-3-phosphate + Dihydroxyacetone phosphate
• Enzyme: Aldolase
5. Dihydroxyacetone phosphate ↔ Glyceraldehyde-3-phosphate
• Enzyme: Triose phosphate isomerase
6. Glyceraldehyde-3-phosphate → 1,3-Bisphosphoglycerate
• Enzyme: Glyceraldehyde-3-phosphate dehydrogenase
• NAD⁺ reduced to NADH
 7. 1,3-Bisphosphoglycerate → 3-Phosphoglycerate
• Enzyme: Phosphoglycerate kinase
• ATP produced (substrate-level phosphorylation)
8. 3-Phosphoglycerate → 2-Phosphoglycerate
• Enzyme: Phosphoglycerate mutase
9. 2-Phosphoglycerate → Phosphoenolpyruvate
• Enzyme: Enolase
10.Phosphoenolpyruvate → Pyruvate
• Enzyme: Pyruvate kinase
• ATP produced (substrate-level phosphorylation)
11.Pyruvate → Lactate
• Enzyme: Lactate dehydrogenase
• NADH oxidized to NAD⁺ (regenerates NAD⁺ for glycolysis)1
Net Reaction:
Glucose+2 ADP+2 Pi→2 Lactate+2 ATP+2 H2OGlucose+2 ADP+2 Pi→2 Lactate+2 ATP+2 H2O
Energy Yield
• Net gain: 2 ATP per glucose (no net NADH gain, as NADH is reoxidized to NAD⁺ during lactate formation)1.
Biological Role of Anaerobic Glycolysis
• ATP production without oxygen: Provides rapid, though limited, ATP supply when oxygen is scarce or mitochondria are
absent.
 • Critical for certain tissues: Main or sole energy source for erythrocytes, exercising muscle, and some neural tissues.
• Lactate as a shuttle: Lactate produced can be transported to the liver (Cori cycle) and converted back to glucose via
gluconeogenesis.
• Survival during hypoxia: Enables tissues to survive temporary oxygen deprivation.
• Clinical relevance: Increased anaerobic glycolysis is seen in ischemia, shock, intense exercise, and many tumors
(Warburg effect)1.

88. Sources and ways of using lactic acid.

Ans. Sources of Lactic Acid
Lactic acid (lactate) is produced in the body primarily through the process of anaerobic glycolysis:
• Skeletal Muscle: During intense exercise or oxygen deficiency, pyruvate (from glycolysis) is reduced to lactate by lactate
dehydrogenase to regenerate NAD⁺, allowing glycolysis to continue5.
• Erythrocytes: Red blood cells lack mitochondria and always convert glucose to lactate, even under aerobic conditions5.
• Tissues with Low Oxygen Supply: Such as the skin, renal medulla, lens, cornea, and some brain regions, also produce
lactate.
• Rapidly Growing Tumors: Cancer cells often rely on anaerobic glycolysis (Warburg effect), leading to high lactate
production.
• Other Tissues: Under hypoxic or ischemic conditions, many tissues increase lactate production.
Ways of Using Lactic Acid
Lactic acid is not simply a waste product; it plays several important metabolic roles:
1. Conversion to Pyruvate (Oxidation)
 • Aerobic Tissues: When oxygen becomes available, lactate is transported to tissues with high oxidative capacity (heart,
liver, kidneys, skeletal muscle at rest) and is converted back to pyruvate by lactate dehydrogenase. Pyruvate then enters
the TCA cycle for complete oxidation to CO₂ and H₂O, generating ATP5.
2. Gluconeogenesis (Cori Cycle)
• Liver (mainly) and Kidneys: Lactate produced in peripheral tissues is transported via the bloodstream to the liver, where
it is converted back to glucose through gluconeogenesis (the Cori cycle). This glucose can then be released into the
bloodstream and reused by muscles and other tissues5.
• Cori Cycle:
Muscle glucose → pyruvate → lactate → blood → liver → pyruvate → glucose → blood → muscle.
3. Fuel for the Heart
• Cardiac Muscle: The heart efficiently utilizes lactate as an energy source, especially during exercise, converting it to
pyruvate and oxidizing it in the TCA cycle.
4. Other Uses
• Substrate for Amino Acid Synthesis: Pyruvate derived from lactate can be transaminated to form alanine.
• Maintenance of Redox Balance: The interconversion of pyruvate and lactate helps maintain the NAD⁺/NADH ratio in the
cytosol, which is essential for continued glycolysis under anaerobic conditions5.

89. Methods for the quantitative determination of lactate in biological fluids. Reference values, diagnostic value.
Ans. Methods of Determination
1. Enzymatic Methods (Lactate Oxidase or Lactate Dehydrogenase-Based)
• Lactate oxidase method:
 • Lactate is oxidized by lactate oxidase to pyruvate and hydrogen peroxide. The hydrogen peroxide is then detected
colorimetrically using a chromogenic substrate (e.g., peroxidase + a dye), and the intensity of the color is
proportional to lactate concentration.
• Lactate dehydrogenase (LDH) method:
• Lactate is converted to pyruvate by LDH, with simultaneous reduction of NAD⁺ to NADH. The increase in NADH is
measured spectrophotometrically at 340 nm and is directly proportional to lactate concentration1.
2. Spectrophotometric Methods
• Both enzymatic reactions can be monitored by measuring changes in absorbance at specific wavelengths, allowing for
quantitative analysis of lactate in plasma, serum, or other fluids1.
3. Point-of-Care and Automated Analyzers
• Many modern clinical laboratories and intensive care units use automated analyzers or handheld devices for rapid
lactate measurement, often based on enzymatic or biosensor technology.
Reference Values
• Normal blood lactate (venous):
0.5–2.2 mmol/L (may vary slightly by laboratory and sample type)
• Elevated levels:
• 2.2 mmol/L: Hyperlactatemia
• 4.0 mmol/L: Lactic acidosis (clinically significant)
Diagnostic Value
• Lactate is a key marker of tissue hypoxia, anaerobic metabolism, and impaired oxidative phosphorylation.
• Clinical uses:
 • Diagnosis and monitoring of lactic acidosis: Seen in shock, sepsis, severe hypoxia, cardiac or respiratory failure.
• Assessment of tissue perfusion: Elevated lactate indicates inadequate oxygen delivery or utilization.
• Prognostic marker: High lactate levels are associated with increased mortality in critically ill patients.
• Monitoring therapy: Used to assess response to interventions in shock or sepsis.
• Other uses:
• Detection of metabolic disorders, mitochondrial diseases, and some poisonings.

90. Aerobic breakdown of glucose: localization in the cell, stages of the process, the sequence of reactions of the anaerobic
stage.
Ans. Cellular Localization
• Glycolysis (Anaerobic Stage): Occurs in the cytosol of all cells.
• Pyruvate Oxidation, Citric Acid Cycle, and Electron Transport Chain (Aerobic Stages): Occur in the mitochondria.
Stages of Aerobic Glucose Oxidation
1. Glycolysis (Anaerobic Stage)
• Glucose (6C) is converted to two molecules of pyruvate (3C) through a series of ten enzyme-catalyzed reactions in
the cytosol.
• Net yield: 2 ATP (substrate-level phosphorylation) and 2 NADH per glucose.
2. Oxidative Decarboxylation of Pyruvate
• Each pyruvate is transported into the mitochondrial matrix and converted to acetyl-CoA by the pyruvate
dehydrogenase complex.
 • Products: 1 NADH and 1 CO₂ per pyruvate.
3. Citric Acid Cycle (Krebs Cycle)
• Acetyl-CoA enters the cycle and is oxidized to 2 CO₂.
• Each turn produces 3 NADH, 1 FADH₂, and 1 GTP (or ATP) per acetyl-CoA.
4. Electron Transport Chain and Oxidative Phosphorylation
• NADH and FADH₂ donate electrons to the electron transport chain (ETC) in the inner mitochondrial membrane.
• Electrons flow through complexes I-IV, reducing O₂ to H₂O.
• The energy released pumps protons, creating a proton gradient used by ATP synthase to produce ATP.
• Total ATP yield: Up to 36–38 ATP per glucose (depending on shuttle systems and cell type)145.
Sequence of Reactions in the Anaerobic Stage (Glycolysis)
Preparatory Phase:
1. Phosphorylation of Glucose
Glucose + ATP → Glucose-6-phosphate (Hexokinase)
2. Isomerization
Glucose-6-phosphate → Fructose-6-phosphate (Phosphoglucose isomerase)
3. Second Phosphorylation
Fructose-6-phosphate + ATP → Fructose-1,6-bisphosphate (Phosphofructokinase-1)
4. Cleavage
Fructose-1,6-bisphosphate → Glyceraldehyde-3-phosphate + Dihydroxyacetone phosphate (Aldolase)
5. Isomerization of DHAP
Dihydroxyacetone phosphate ↔ Glyceraldehyde-3-phosphate (Triose phosphate isomerase)
Payoff Phase (each reaction occurs twice per glucose):
6. Oxidation and Phosphorylation
Glyceraldehyde-3-phosphate + NAD⁺ + Pi → 1,3-Bisphosphoglycerate + NADH + H⁺ (Glyceraldehyde-3-phosphate
dehydrogenase)
7. Substrate-level Phosphorylation
1,3-Bisphosphoglycerate + ADP → 3-Phosphoglycerate + ATP (Phosphoglycerate kinase)
8. Mutase Reaction
3-Phosphoglycerate → 2-Phosphoglycerate (Phosphoglycerate mutase)
9. Dehydration
2-Phosphoglycerate → Phosphoenolpyruvate (Enolase)
10. Substrate-level Phosphorylation
Phosphoenolpyruvate + ADP → Pyruvate + ATP (Pyruvate kinase)
Net Reaction of Glycolysis:
Glucose+2 NAD++2 ADP+2 Pi→2 Pyruvate+2 NADH+2 ATP+2 H2O+2 H+Glucose+2 NAD++2 ADP+2 Pi→2 Pyruvate+2 NADH+2 A
TP+2 H2O+2 H+
91. Pyruvate. Sources. The role of the pyruvate dehydrogenase complex in providing the tricarboxylic acid cycle with
substrates.
Ans. Sources of Pyruvate
Pyruvate is a central metabolic intermediate formed primarily through:
• Glycolysis: The main pathway, where one molecule of glucose is converted into two molecules of pyruvate in the cytosol
of all cells2.
• Amino Acid Catabolism: Several glucogenic amino acids, such as alanine (by transamination), serine, glycine, cysteine,
threonine, and tryptophan, can be converted into pyruvate during their catabolism2.
 • Lactate Oxidation: Lactate, produced from anaerobic glycolysis, can be converted back to pyruvate by lactate
dehydrogenase2.
• Other Pathways: Pyruvate can also be generated from the decarboxylation of oxaloacetate and from the metabolism of
some dicarboxylic acids2.
Role of the Pyruvate Dehydrogenase Complex (PDH) in Supplying the TCA Cycle
Function and Reaction
• The pyruvate dehydrogenase complex (PDH) is a large, multi-enzyme complex located in the mitochondrial matrix4.
• It catalyzes the irreversible oxidative decarboxylation of pyruvate to acetyl-CoA, linking glycolysis to the tricarboxylic
acid (TCA, Krebs) cycle24:
Pyruvate+CoA-SH+NAD+→Acetyl-CoA+CO2+NADH+H+Pyruvate+CoA-SH+NAD+→Acetyl-CoA+CO2+NADH+H+
• Acetyl-CoA produced by this reaction is the essential two-carbon substrate that enters the TCA cycle by condensing with
oxaloacetate to form citrate, initiating the cycle24.
Cofactors and Vitamins Required
The PDH complex requires five cofactors, each derived from essential vitamins:
• Thiamine pyrophosphate (TPP) (from vitamin B1)1
• Lipoic acid
• Coenzyme A (from pantothenic acid, vitamin B5)
• FAD (from riboflavin, vitamin B2)
• NAD⁺ (from niacin, vitamin B3)
Significance in Metabolism
 • Gateway to Aerobic Metabolism: The PDH complex is the critical step that commits pyruvate (and thus glucose and
many amino acids) to aerobic oxidation in the mitochondria, providing acetyl-CoA for the TCA cycle and subsequent ATP
production via oxidative phosphorylation24.
• Regulation: The activity of PDH is tightly regulated by energy status (inhibited by high ATP, NADH, and acetyl-CoA;
activated by ADP, NAD⁺, and CoA-SH), ensuring acetyl-CoA is produced according to cellular energy needs4.
• Clinical Relevance: Deficiency or inhibition of PDH (e.g., by thiamine deficiency) impairs aerobic energy production,
leading to lactic acidosis and serious neurological symptoms1.

92. The Krebs cycle is a common final pathway of oxidation of metabolic products of carbohydrates, fatty acids and amino
acids. Energy balance of aerobic glucose breakdown.
Ans. The Krebs Cycle as a Metabolic Hub
The Krebs cycle (also called the citric acid cycle or tricarboxylic acid cycle, TCA) is the central pathway where the oxidative
catabolism of carbohydrates, fatty acids, and amino acids converges. Regardless of the original nutrient:
• Carbohydrates are broken down to pyruvate via glycolysis, then converted to acetyl-CoA.
• Fatty acids undergo β-oxidation to yield acetyl-CoA1.
• Amino acids are deaminated and their carbon skeletons are converted to intermediates of the TCA cycle or acetyl-CoA.
All these acetyl-CoA molecules enter the Krebs cycle, where their acetyl groups are fully oxidized to CO₂, generating reduced
coenzymes (NADH, FADH₂) that drive ATP synthesis in the electron transport chain246.
Stages of Aerobic Glucose Breakdown
1. Glycolysis (Cytosol)
• Glucose → 2 Pyruvate
 • Net yield: 2 ATP (substrate-level phosphorylation), 2 NADH
2. Oxidative Decarboxylation of Pyruvate (Mitochondria)
• Pyruvate → Acetyl-CoA + CO₂ + NADH (via the pyruvate dehydrogenase complex)
3. Krebs Cycle (Mitochondrial Matrix)
• Acetyl-CoA (2C) + Oxaloacetate (4C) → Citrate (6C)
• Through a series of reactions, citrate is converted back to oxaloacetate, releasing 2 CO₂, and generating:
• 3 NADH
• 1 FADH₂
• 1 GTP (or ATP)
• 2 CO₂ (per acetyl-CoA)
4. Electron Transport Chain & Oxidative Phosphorylation (Inner Mitochondrial Membrane)
• NADH and FADH₂ donate electrons to the ETC, reducing O₂ to H₂O and creating a proton gradient that drives ATP
synthesis.
Energy Balance of Complete Aerobic Glucose Oxidation
Per 1 molecule of glucose:

Stage ATP (or NADH FADH₂ ATP Yield (approx.)

GTP)

Glycolysis 2 2 0 2 + (2 × 2.5) = 7
 Stage ATP (or NADH FADH₂ ATP Yield (approx.)
GTP)

Pyruvate → Acetyl-
CoA 0 2 0 2 × 2.5 = 5

2 + (6 × 2.5) + (2 × 1.5) = 2 + 15 + 3 =
Krebs Cycle (2 turns) 2 (GTP) 6 2 20

Total 4 10 2 32 ATP

• NADH: Each yields ~2.5 ATP via the ETC.

• FADH₂: Each yields ~1.5 ATP via the ETC.
Grand Total:
• 32–38 ATP per glucose (variation depends on shuttle systems and cell type)24.
• If the glycerol phosphate shuttle is used (muscle), yield is ~36 ATP.
• If the malate-aspartate shuttle is used (liver, heart), yield is up to 38 ATP.
Summary: Why the Krebs Cycle Is the Common Final Pathway
• Carbohydrates, fatty acids, and amino acids are all funneled into the Krebs cycle as acetyl-CoA or as cycle
intermediates.
• The cycle completely oxidizes these substrates to CO₂, generating high-energy electrons stored in NADH and FADH₂.
• These electrons are used in the electron transport chain to produce the majority of cellular ATP.
• The cycle also supplies intermediates for biosynthetic (anabolic) pathways, making it amphibolic26.
Key Points
• Krebs cycle is the metabolic crossroad for energy production from all major nutrients.
• Energy yield from aerobic glucose oxidation: up to 32–38 ATP per glucose molecule.
• The cycle is tightly regulated by energy status (ATP/ADP, NADH/NAD⁺ ratios) and substrate availability246.

93. Pentose phosphate pathway of glucose oxidation. Prevalence in the body, localization. Role in providing anabolic processes
in the body, detoxification of xenobiotics.
Ans. Overview and Localization
The pentose phosphate pathway (PPP), also known as the phosphogluconate pathway or hexose monophosphate (HMP)
shunt, is an alternative oxidative pathway for glucose-6-phosphate metabolism. It operates in the cytosol of cells and consists
of two phases:
• Oxidative phase: Produces NADPH and ribulose-5-phosphate.
• Non-oxidative phase: Interconverts sugars of various lengths, producing ribose-5-phosphate and recycling intermediates
back to glycolysis.
Prevalence in the Body
• The PPP is active in all cells, but especially prominent in tissues with high anabolic and detoxification demands:
• Liver
• Adipose tissue
• Adrenal cortex
• Mammary glands
 • Testes and ovaries
• Erythrocytes (red blood cells)
• Lens and cornea of the eye
• Rapidly dividing cells (e.g., bone marrow, skin, intestinal mucosa)1.
It is less active in skeletal muscle, where glycolysis is the dominant pathway1.
Sequence and Key Reactions
Oxidative Phase
1. Glucose-6-phosphate dehydrogenase (G6PD):
• Glucose-6-phosphate + NADP⁺ → 6-phosphoglucono-δ-lactone + NADPH
2. Lactonase:
• 6-phosphoglucono-δ-lactone + H₂O → 6-phosphogluconate
3. 6-phosphogluconate dehydrogenase:
• 6-phosphogluconate + NADP⁺ → ribulose-5-phosphate + CO₂ + NADPH
End products: 2 NADPH, 1 ribulose-5-phosphate, 1 CO₂ per glucose-6-phosphate oxidized1.
Non-Oxidative Phase
• Transketolase and transaldolase (require thiamine pyrophosphate) catalyze the interconversion of pentose phosphates
and glycolytic intermediates (fructose-6-phosphate, glyceraldehyde-3-phosphate)1.
Role in Anabolic Processes
1. Generation of NADPH:
• NADPH is a key electron donor for reductive biosynthesis (anabolism), including:
 • Fatty acid synthesis (liver, adipose, lactating mammary gland)
• Cholesterol and steroid hormone synthesis (adrenal cortex, gonads)
• Synthesis of some amino acids and nucleotides13.
2. Provision of Ribose-5-Phosphate:
• Ribose-5-phosphate is required for nucleotide and nucleic acid synthesis (DNA, RNA, ATP, NAD⁺, FAD, CoA), especially in
rapidly dividing cells1.
Role in Detoxification of Xenobiotics
1. NADPH for Antioxidant Defense:
• In erythrocytes and the lens, NADPH maintains reduced glutathione (GSH) via glutathione reductase. GSH is essential
for neutralizing hydrogen peroxide and other reactive oxygen species, protecting cells from oxidative damage13.
• G6PD deficiency impairs this defense, leading to hemolytic anemia under oxidative stress.
2. NADPH for Cytochrome P450 System:
• In the liver (microsomal system), NADPH provides reducing equivalents for cytochrome P450 monooxygenases, which
hydroxylate and detoxify drugs, hormones, and other xenobiotics, making them more water-soluble for excretion3.
3. NADPH in Phagocytosis:
• In leukocytes, NADPH is used by NADPH oxidase to generate superoxide radicals and hydrogen peroxide during the
respiratory burst, which are essential for killing engulfed pathogens3.

94. Exogenous and endogenous sources of glucose, ways of using glucose in the body.
Ans. Exogenous Sources of Glucose
 • Dietary Carbohydrates: The primary exogenous source of glucose is the digestion of dietary carbohydrates, including:
• Polysaccharides: Starch (from grains, potatoes, legumes) and glycogen (from animal sources)
• Disaccharides: Sucrose (table sugar), lactose (milk sugar), maltose (from starch digestion)
• Monosaccharides: Glucose, fructose, galactose (in fruits, honey, some vegetables)
• Digestion and Absorption: These carbohydrates are broken down by digestive enzymes into monosaccharides, mainly
glucose, which is absorbed in the small intestine and enters the bloodstream3.
Endogenous Sources of Glucose
• Glycogenolysis: Breakdown of glycogen stored in the liver (and, to a lesser extent, in muscle) releases glucose into the
blood, especially between meals and during fasting13.
• Gluconeogenesis: Synthesis of glucose from non-carbohydrate precursors, mainly in the liver (and kidney), including:
• Amino acids: Especially alanine and glutamine (from muscle protein breakdown)
• Lactate: Produced by anaerobic glycolysis in muscles and red blood cells (Cori cycle)
• Glycerol: From triglyceride breakdown in adipose tissue
• Propionate: Particularly important in ruminants
• Other Pathways: Some gluconeogenesis also occurs from intermediates such as pyruvate and certain TCA cycle
metabolites13.
Ways of Using Glucose in the Body
Glucose is a central metabolic substrate with several key fates:
1. Energy Production
 • Glycolysis: Glucose is converted to pyruvate, generating ATP and NADH. Under anaerobic conditions, pyruvate is
reduced to lactate3.
• Aerobic Oxidation: Pyruvate enters mitochondria, is converted to acetyl-CoA, and is fully oxidized in the TCA (Krebs)
cycle, producing CO₂, water, and large amounts of ATP via oxidative phosphorylation36.
• Pentose Phosphate Pathway: Glucose-6-phosphate is oxidized to produce NADPH (for reductive biosynthesis and
antioxidant defense) and ribose-5-phosphate (for nucleotide synthesis)3.
2. Storage
• Glycogenesis: Glucose is stored as glycogen in the liver (for blood glucose regulation) and muscle (for local energy use).
Glycogen can be rapidly mobilized when needed13.
3. Biosynthesis
• Precursor for Other Molecules: Glucose provides carbon skeletons for the synthesis of:
• Non-essential amino acids (via glycolytic and TCA intermediates)
• Fatty acids and triglycerides (via acetyl-CoA, especially in liver and adipose tissue)
• Nucleotides (via the pentose phosphate pathway)
• Glycoproteins and glycolipids (for cell membranes and signaling)
4. Other Specialized Functions
• Lactate Production and Recycling: In tissues with low oxygen (e.g., red blood cells, exercising muscle), glucose is
converted to lactate, which is then recycled to glucose in the liver (Cori cycle)3.
• Brain Function: Glucose is the primary and essential energy source for the brain under normal conditions6.
• Red Blood Cells: Glucose is the exclusive energy source for erythrocytes, as they lack mitochondria3.
95. Hormonal regulation of blood glucose levels.
Ans. Blood glucose homeostasis is tightly regulated by a complex interplay of hormones, ensuring a stable supply of glucose to
tissues-especially the brain-despite fluctuations in dietary intake, fasting, or stress. The main hormones involved
are insulin, glucagon, adrenaline (epinephrine), glucocorticoids (cortisol), growth hormone, and thyroid hormones.
1. Insulin (Pancreatic β-Cells)
Role: The only hormone that lowers blood glucose.
• Stimulus: Secreted in response to elevated blood glucose (postprandial state).
• Mechanism of action: Binds to cell surface receptors, triggering the translocation of GLUT4 transporters to the
membrane in muscle and adipose tissue, increasing glucose uptake. Activates key enzymes for glycolysis, glycogenesis,
and lipogenesis, and inhibits gluconeogenesis and glycogenolysis23.
• Metabolic effects:
• Increases glucose uptake (muscle, adipose)
• Stimulates glycogen synthesis (liver, muscle)
• Enhances glycolysis and fatty acid synthesis
• Inhibits glycogen breakdown and gluconeogenesis
• Promotes protein synthesis
2. Glucagon (Pancreatic α-Cells)
Role: Main hormone that raises blood glucose during fasting.
• Stimulus: Secreted in response to low blood glucose (fasting, between meals).
• Mechanism of action: Binds to G protein-coupled receptors on hepatocytes, activating adenylate cyclase and increasing
cAMP, which activates protein kinase A and stimulates glycogenolysis and gluconeogenesis23.
 • Metabolic effects:
• Stimulates glycogen breakdown (glycogenolysis) in the liver
• Activates gluconeogenesis (formation of glucose from non-carbohydrate sources)
• Inhibits glycogen synthesis and glycolysis in the liver
• Promotes lipolysis in adipose tissue
3. Adrenaline (Epinephrine, Adrenal Medulla)
Role: Rapidly increases blood glucose during acute stress ("fight or flight" response).
• Stimulus: Stress, hypoglycemia, exercise.
• Mechanism of action: Acts via adrenergic receptors, activating cAMP pathways in liver and muscle, stimulating
glycogenolysis; also stimulates glucagon secretion and inhibits insulin release23.
• Metabolic effects:
• Promotes glycogenolysis in liver and muscle
• Stimulates gluconeogenesis in the liver
• Inhibits insulin secretion, stimulates glucagon release
• Increases lipolysis in adipose tissue
4. Glucocorticoids (Cortisol, Adrenal Cortex)
Role: Increases blood glucose during prolonged stress and fasting.
• Stimulus: Stress, ACTH stimulation.
• Mechanism of action: Binds to intracellular receptors, modulating gene expression to increase gluconeogenic enzymes
and reduce glucose utilization by peripheral tissues23.
 • Metabolic effects:
• Stimulates gluconeogenesis in the liver (from amino acids and glycerol)
• Increases protein catabolism in muscle (providing amino acids for gluconeogenesis)
• Reduces glucose uptake/utilization in peripheral tissues ("anti-insulin effect")
• Promotes lipolysis
5. Growth Hormone (Anterior Pituitary)
Role: Increases blood glucose, especially during growth and fasting.
• Mechanism of action: Reduces glucose uptake in tissues, increases lipolysis, and stimulates gluconeogenesis13.
• Metabolic effects:
• Decreases glucose uptake in muscle and adipose tissue
• Increases lipolysis (fat breakdown)
• Stimulates gluconeogenesis
6. Thyroid Hormones (T3, T4)
Role: Modulate carbohydrate metabolism, generally increasing blood glucose.
• Mechanism of action: Increase basal metabolic rate, stimulate glycogenolysis, gluconeogenesis, and intestinal glucose
absorption13.
• Metabolic effects:
• Enhance glucose absorption from the gut
• Stimulate glycogenolysis and gluconeogenesis
• Increase insulin degradation
96. Physiological and pathological hyperglycemia, hypoglycemia, causes. Diabetes mellitus and steroid. Typical metabolic
disorders in diabetes. Diagnostics
Ans. Physiological and Pathological Hyperglycemia
Physiological Hyperglycemia
• Definition: A temporary, moderate increase in blood glucose that occurs in normal physiological states.
• Causes:
• After meals (postprandial): Due to absorption of dietary carbohydrates.
• Stress, excitement, pain, or physical exertion: Due to increased secretion of counter-regulatory hormones
(adrenaline, cortisol, growth hormone).
• Pregnancy: Mild increase due to hormonal changes.
• Mechanism: Rapidly corrected by increased insulin secretion and enhanced glucose uptake/storage.
Pathological Hyperglycemia
• Definition: Persistent elevation of blood glucose above normal fasting levels (typically >7.0 mmol/L fasting; >11.1
mmol/L at any time).
• Causes:
• Diabetes mellitus (most common)
• Endocrine disorders: Cushing's syndrome (excess glucocorticoids), acromegaly (excess growth hormone),
pheochromocytoma (excess catecholamines), thyrotoxicosis (excess thyroid hormones).
• Pancreatic diseases: Pancreatitis, pancreatic tumors.
 • Liver disease: Impaired glucose metabolism.
• Drugs: Glucocorticoids, thiazide diuretics, some antipsychotics.
Physiological and Pathological Hypoglycemia
Physiological Hypoglycemia
• Definition: A transient drop in blood glucose, usually mild and self-limited.
• Causes:
• Fasting or prolonged exercise: Increased glucose utilization.
• Neonates and infants: Higher metabolic rate and lower glycogen reserves.
Pathological Hypoglycemia
• Definition: Abnormally low blood glucose (<3.0 mmol/L in adults), which can cause neuroglycopenic symptoms.
• Causes:
• Insulin excess: Insulinoma, overdose of insulin or oral hypoglycemics.
• Severe liver disease: Impaired gluconeogenesis/glycogenolysis.
• Adrenal insufficiency: Cortisol deficiency.
• Hypopituitarism: Deficiency of counter-regulatory hormones.
• Sepsis, renal failure, malnutrition, alcohol abuse.
Diabetes Mellitus and Steroid (Secondary) Diabetes
Diabetes Mellitus
• Definition: A group of chronic metabolic diseases characterized by persistent hyperglycemia due to defects in insulin
secretion, insulin action, or both.
 • Types:
• Type 1 (IDDM): Absolute insulin deficiency due to autoimmune destruction of pancreatic β-cells. Early onset,
rapid progression.
• Type 2 (NIDDM): Insulin resistance and relative insulin deficiency. Associated with obesity, older age, slow
progression.
• Other specific types: Genetic defects, pancreatic diseases, endocrinopathies, drug-induced.
Steroid (Secondary) Diabetes
• Definition: Hyperglycemia/diabetes caused by chronic exposure to glucocorticoids (endogenous or exogenous).
• Mechanism: Glucocorticoids increase gluconeogenesis, decrease peripheral glucose uptake, and antagonize insulin
action, leading to hyperglycemia5.
Typical Metabolic Disorders in Diabetes Mellitus
Carbohydrate Metabolism
• Decreased glucose uptake by insulin-dependent tissues (muscle, adipose).
• Increased gluconeogenesis and glycogenolysis in the liver.
• Persistent hyperglycemia and glycosuria (glucose in urine).
• Polyuria, polydipsia, polyphagia (classic triad).
Lipid Metabolism
• Increased lipolysis in adipose tissue (due to low insulin), leading to elevated free fatty acids in plasma.
• Increased hepatic ketogenesis: Excess acetyl-CoA is converted to ketone bodies, leading to ketoacidosis (mainly in type
1 DM).
 • Hypertriglyceridemia and increased risk of atherosclerosis.
Protein Metabolism
• Increased proteolysis: Muscle wasting and weight loss.
• Negative nitrogen balance.
Electrolyte and Water Balance
• Dehydration: Due to osmotic diuresis from glycosuria.
• Electrolyte imbalances: Loss of sodium, potassium, and other ions in urine.
Diagnostics of Diabetes Mellitus
Diagnostic Criteria (WHO/ADA)
• Fasting plasma glucose (FPG): ≥7.0 mmol/L (126 mg/dL)
• 2-hour plasma glucose (OGTT): ≥11.1 mmol/L (200 mg/dL) after 75 g oral glucose
• Random plasma glucose: ≥11.1 mmol/L (200 mg/dL) with symptoms
• HbA1c: ≥6.5% (48 mmol/mol)
Other Laboratory Findings
• Glycosuria: Glucose in urine (when blood glucose >10 mmol/L, renal threshold).
• Ketonuria: Presence of ketone bodies in urine (type 1 DM, diabetic ketoacidosis).
• Elevated plasma triglycerides and cholesterol.
• Decreased C-peptide (type 1 DM).
Additional Tests
 • Oral glucose tolerance test (OGTT): For borderline or gestational diabetes.
• Autoantibodies: (Type 1 DM) e.g., anti-GAD, islet cell antibodies.
• Urine microalbumin: Early marker of diabetic nephropathy.

97. Blood glucose, sources, reference values. Methods for the quantitative determination of glucose.
Ans. Sources of Blood Glucose
1. Exogenous Sources
• Dietary carbohydrates: The primary source, including starches, disaccharides (sucrose, lactose), and monosaccharides
(glucose, fructose, galactose), which are digested and absorbed in the small intestine as glucose7.
• Absorption: After digestion, glucose enters the bloodstream via the portal vein.
2. Endogenous Sources
• Glycogenolysis: Breakdown of glycogen stored in the liver releases glucose into the blood, especially during fasting or
between meals3.
• Gluconeogenesis: Synthesis of glucose from non-carbohydrate precursors (amino acids, lactate, glycerol) in the liver
and, to a lesser extent, in the kidneys3.
• Conversion from other monosaccharides: Galactose and fructose (from dietary sources) can be converted to glucose in
the liver7.
Reference Values for Blood Glucose
• Fasting blood glucose (venous plasma):
3.3–5.5 mmol/L (60–99 mg/dL) is considered the normal reference range for adults37.
 • Postprandial (2 hours after eating):
Should not exceed 7.8 mmol/L (140 mg/dL).
• Impaired fasting glucose:
5.6–6.9 mmol/L (100–125 mg/dL).
• Diabetes mellitus (diagnostic threshold):
Fasting ≥7.0 mmol/L (≥126 mg/dL), or random/postprandial ≥11.1 mmol/L (≥200 mg/dL)3.
Methods for the Quantitative Determination of Glucose
1. Enzymatic Methods (Most Common in Clinical Practice)
• Glucose Oxidase Method:
• Principle: Glucose is oxidized by glucose oxidase to gluconic acid and hydrogen peroxide. The hydrogen peroxide
reacts with a chromogen in the presence of peroxidase, producing a colored compound measured
spectrophotometrically.
• Advantages: High specificity, sensitivity, and widely used in automated analyzers.
• Hexokinase Method:
• Principle: Glucose is phosphorylated by hexokinase and ATP to form glucose-6-phosphate, which is then oxidized
by glucose-6-phosphate dehydrogenase, reducing NAD(P)⁺ to NAD(P)H. The increase in absorbance at 340 nm is
proportional to glucose concentration.
• Advantages: Highly specific, considered the reference method in many laboratories.
2. Chemical Methods (Less Common Today)
• Folin-Wu, Somogyi-Nelson, and o-toluidine methods:
• Based on reduction of copper or formation of colored complexes with glucose.
 • Limitations: Prone to interference from other reducing substances; less specific than enzymatic methods.
3. Point-of-Care and Bedside Testing
• Glucometers:
• Use glucose oxidase or dehydrogenase-based test strips for rapid, semi-quantitative measurement of capillary
blood glucose (fingerstick).
• Essential for diabetes self-monitoring.

98. Gluconeogenesis. Connection with transamination, sequence of reactions. Hormonal regulation.

Ans. What Is Gluconeogenesis?
Gluconeogenesis is the metabolic process by which glucose is synthesized from non-carbohydrate precursors.
• Main sites: Liver (major) and kidney cortex (minor, more prominent during prolonged fasting).
Key substrates:
• Lactate (from anaerobic glycolysis, mainly muscle and red blood cells)
• Amino acids (especially alanine and glutamine)
• Glycerol (from triglyceride breakdown)
• Propionate (in ruminants)
Connection with Transamination
• Transamination reactions convert amino acids into α-keto acids.
• Glucogenic amino acids (e.g., alanine, glutamine) are converted to intermediates (pyruvate, oxaloacetate) that feed
directly into the gluconeogenic pathway.
 • Key example:
• Alanine + α-ketoglutarate ⇄ pyruvate + glutamate
• (Catalyzed by alanine aminotransferase, with pyridoxal phosphate as a coenzyme.)
Sequence of Key Reactions in Gluconeogenesis
Start with pyruvate, lactate, or an amino acid-derived intermediate:
1. Pyruvate Carboxylation
• Pyruvate + CO₂ + ATP → Oxaloacetate + ADP + Pi
• Enzyme: Pyruvate carboxylase (mitochondria; requires biotin)
2. Oxaloacetate to Phosphoenolpyruvate (PEP)
• Oxaloacetate + GTP → PEP + CO₂ + GDP
• Enzyme: PEP carboxykinase (mitochondria or cytosol)
3. Reverse Glycolysis to Fructose-1,6-bisphosphate
• PEP is converted by glycolytic enzymes (in reverse) through 2-phosphoglycerate, 3-phosphoglycerate, 1,3-
bisphosphoglycerate, glyceraldehyde-3-phosphate, and fructose-1,6-bisphosphate.
4. Dephosphorylation of Fructose-1,6-bisphosphate
• Fructose-1,6-bisphosphate → Fructose-6-phosphate + Pi
• Enzyme: Fructose-1,6-bisphosphatase (key regulated step)
5. Isomerization and Dephosphorylation of Glucose-6-phosphate
• Fructose-6-phosphate → Glucose-6-phosphate (phosphohexose isomerase)
• Glucose-6-phosphate → Glucose + Pi
 • Enzyme: Glucose-6-phosphatase (only in liver/kidney)
Glycerol and Other Substrates
• Glycerol → Glycerol-3-phosphate → Dihydroxyacetone phosphate (DHAP)
• DHAP enters the pathway at the triose phosphate stage.
Hormonal Regulation of Gluconeogenesis
• Insulin:
• Inhibits gluconeogenesis.
• Suppresses the expression/activity of key gluconeogenic enzymes (PEP carboxykinase, fructose-1,6-
bisphosphatase, glucose-6-phosphatase).
• Glucagon:
• Stimulates gluconeogenesis (mainly in the liver).
• Increases cAMP/PKA signaling, activating gene transcription of gluconeogenic enzymes and inhibiting glycolytic
enzymes.
• Cortisol (glucocorticoids):
• Stimulates gluconeogenesis by increasing the supply of substrates (via proteolysis and amino acid mobilization)
and upregulating gluconeogenic enzymes.
• Adrenaline:
• Increases gluconeogenesis during stress (stimulates glycogenolysis and provides substrates for gluconeogenesis).

99. Ways of formation and use of pyruvic acid and acetyl-CoA in the body. The value of processes.
Ans. Formation of Pyruvic Acid (Pyruvate)
Pyruvate is a central intermediate in metabolism and is formed in the body by several key pathways:
• Glycolysis: The main source; glucose is converted to two molecules of pyruvate through a series of cytosolic reactions in
virtually all tissues. This is the endpoint of glycolysis under both aerobic and anaerobic conditions1.
• Amino Acid Catabolism: Several glucogenic amino acids (e.g., alanine, serine, glycine, cysteine, threonine) are converted
to pyruvate during their breakdown. Alanine, for example, is transaminated to pyruvate1.
• Lactate Oxidation: Lactate, produced by anaerobic glycolysis (especially in muscle and red blood cells), is converted back
to pyruvate by lactate dehydrogenase1.
• Decarboxylation of Oxaloacetate: Oxaloacetate can be decarboxylated to pyruvate, either spontaneously or via
oxaloacetate decarboxylase1.
Fates and Uses of Pyruvic Acid
Pyruvate serves as a metabolic crossroad and can be utilized in several ways1:
• Aerobic Conditions:
• Oxidative Decarboxylation to Acetyl-CoA: In the presence of oxygen, pyruvate enters mitochondria and is
converted to acetyl-CoA by the pyruvate dehydrogenase (PDH) complex. This is the main route for energy
production from carbohydrates1.
• Anaerobic Conditions:
• Reduction to Lactate: In the absence of oxygen (e.g., in vigorously contracting muscle or erythrocytes), pyruvate is
reduced to lactate to regenerate NAD⁺, allowing glycolysis to continue1.
• Transamination:
• Formation of Alanine: Pyruvate can accept an amino group to form alanine, which is important in the glucose-
alanine cycle (especially between muscle and liver)1.
 • Gluconeogenesis:
• Formation of Glucose: In the liver and kidney, pyruvate can be used to synthesize glucose via gluconeogenesis,
especially during fasting or starvation1.
• Carboxylation:
• Formation of Oxaloacetate: Pyruvate can be carboxylated to oxaloacetate by pyruvate carboxylase, a key step in
gluconeogenesis and anaplerosis (replenishment of TCA cycle intermediates)1.
Formation of Acetyl-CoA
Acetyl-CoA is a two-carbon molecule that plays a central role in metabolism. It is formed by:
• Oxidative Decarboxylation of Pyruvate:
• Pyruvate Dehydrogenase Complex (PDH): Located in the mitochondrial matrix, this multi-enzyme complex
catalyzes the irreversible conversion of pyruvate to acetyl-CoA, producing NADH and CO₂. This reaction requires
five coenzymes: thiamine pyrophosphate (TPP), lipoic acid, coenzyme A (from pantothenic acid), FAD (from
riboflavin), and NAD⁺ (from niacin)134.
• β-Oxidation of Fatty Acids:
• Fatty acids are broken down in mitochondria to yield acetyl-CoA, which can enter the TCA cycle2.
• Catabolism of Ketogenic Amino Acids:
• Some amino acids (e.g., leucine, lysine) are degraded to acetyl-CoA2.
Fates and Uses of Acetyl-CoA
Acetyl-CoA is a key metabolic intermediate with multiple crucial roles123:
• Entry into the Citric Acid (Krebs) Cycle:
 • Acetyl-CoA condenses with oxaloacetate to form citrate, starting the TCA cycle, which is the main pathway for
energy (ATP) production in aerobic cells14.
• Fatty Acid Synthesis:
• In the cytosol, acetyl-CoA is the precursor for fatty acid and cholesterol synthesis (after being exported from
mitochondria as citrate)2.
• Ketone Body Formation:
• In the liver, excess acetyl-CoA is converted to ketone bodies (acetoacetate, β-hydroxybutyrate, acetone), which
can be used as fuel by extrahepatic tissues during fasting or diabetes2.
• Synthesis of Steroids and Other Compounds:
• Acetyl-CoA is the starting material for cholesterol, steroid hormones, and some neurotransmitters2.
• Acetylation Reactions:
• Acetyl-CoA donates acetyl groups for the acetylation of proteins and other molecules, affecting their function and
regulation3.
Significance of These Processes
• Central Role in Energy Metabolism:
• Both pyruvate and acetyl-CoA are at the crossroads of carbohydrate, fat, and protein metabolism, ensuring
metabolic flexibility and integration12.
• ATP Production:
• The conversion of glucose and fatty acids to acetyl-CoA and its oxidation in the TCA cycle is the main source of
cellular ATP124.
• Biosynthetic Precursor:
 • Pyruvate and acetyl-CoA provide carbon skeletons for the synthesis of amino acids, fatty acids, glucose
(gluconeogenesis), and other essential biomolecules12.
• Metabolic Adaptation:
• These intermediates allow the body to adapt to varying energy demands, fasting, feeding, exercise, and different
physiological states12.
• Clinical Relevance:
• Defects in pyruvate metabolism (e.g., pyruvate dehydrogenase deficiency) or acetyl-CoA utilization (e.g.,
mitochondrial disorders) lead to severe metabolic diseases, lactic acidosis, and energy deficits13.

100. Biological membranes. Chemical structure, properties, functions. Proteins and lipids of membranes.
Ans. Chemical Structure of Biological Membranes
Biological membranes are primarily composed of a lipid bilayer interspersed with various proteins and, in many cases,
carbohydrates attached to lipids and proteins. The main components are:
• Lipids (40–50%):
• Phospholipids (glycerophospholipids and sphingophospholipids) are the primary structural elements, forming a
bilayer with hydrophilic heads facing outward and hydrophobic tails inward.
• Cholesterol is embedded within the bilayer, modulating fluidity and stability.
• Glycolipids (e.g., cerebrosides, gangliosides) are present mainly in the outer leaflet, contributing to cell
recognition and signaling16.
• Proteins (40–60%):
 • Integral (transmembrane) proteins span the bilayer and participate in transport, signaling, and enzymatic
functions.
• Peripheral proteins are attached to membrane surfaces, often involved in signaling or structural support35.
• Carbohydrates:
• Present as oligosaccharide chains covalently linked to proteins (glycoproteins) or lipids (glycolipids), forming the
glycocalyx on the cell surface, which is important for cell recognition and protection5.
Properties of Biological Membranes
• Fluid Mosaic Model: Membranes are dynamic structures where lipids and proteins can move laterally, providing
flexibility and allowing the formation of specialized domains.
• Selective Permeability: The hydrophobic core restricts passage of polar and charged molecules, while allowing diffusion
of small, nonpolar substances. Specific proteins mediate transport of ions and larger molecules.
• Asymmetry: The lipid and protein composition differs between the inner and outer leaflets, crucial for functions like
signaling and membrane trafficking.
• Self-Sealing and Flexibility: The bilayer can reseal if disrupted and allows for membrane fusion, budding, and vesicle
formation.
• Electrical Properties: The membrane can maintain ion gradients and membrane potential, essential for nerve impulse
transmission and muscle contraction6.
Functions of Biological Membranes
• Barrier Function: Separates the cell from its environment and maintains distinct internal compartments (organelles).
• Transport: Regulates the movement of ions, nutrients, and waste products via channels, carriers, and pumps.
• Signal Transduction: Membrane proteins act as receptors for hormones, neurotransmitters, and other signals, initiating
intracellular responses.
 • Cell Recognition and Communication: Glycoproteins and glycolipids are involved in immune recognition, tissue
organization, and cell-cell communication.
• Structural Support and Shape: Anchors the cytoskeleton and extracellular matrix, maintaining cell shape and enabling
movement.
• Energy Conversion: Mitochondrial and chloroplast membranes house the machinery for ATP synthesis and
photosynthesis, respectively26.
Membrane Lipids
• Phospholipids:
• Glycerophospholipids (e.g., phosphatidylcholine, phosphatidylethanolamine, phosphatidylserine) are the main
membrane lipids, forming the bilayer structure.
• Sphingophospholipids (e.g., sphingomyelin) are abundant in nerve tissue and myelin sheaths16.
• Cholesterol:
• Modulates membrane fluidity and mechanical stability, reduces permeability to small molecules, and is a
precursor for steroid hormones and bile acids16.
• Glycolipids:
• Include cerebrosides and gangliosides, especially in nerve tissue, contributing to membrane stability and cell
recognition16.
Membrane Proteins
• Integral (Transmembrane) Proteins:
• Span the lipid bilayer; involved in transport (channels, carriers), signal transduction (receptors), and enzymatic
activity.
 • Peripheral Proteins:
• Loosely attached to the membrane surface; function in signaling, maintaining cell shape, and anchoring the
cytoskeleton35.
• Glycoproteins:
• Proteins with attached carbohydrate chains, important for cell-cell recognition, immune response, and as
receptors5.

101. Classification and properties of lipids in the human body. Representatives, biological role.
Ans. Classification of Lipids
Lipids are a diverse group of hydrophobic or amphipathic organic compounds, unified by their insolubility in water and
solubility in non-polar solvents. The main classes of lipids in the human body are1:
1. Simple Lipids
• Glycerides (Acylglycerols): Esters of glycerol and fatty acids. Includes mono-, di-, and triacylglycerols (triglycerides),
which are the main storage form of fat in adipose tissue.
• Waxes: Esters of long-chain fatty acids with long-chain alcohols. Found in skin, hair, and protective coatings.
2. Complex Lipids
• Phospholipids: Contain fatty acids, alcohol (glycerol or sphingosine), phosphoric acid, and a nitrogenous base (e.g.,
choline, ethanolamine, serine). Subclasses:
• Glycerophospholipids (e.g., phosphatidylcholine, phosphatidylethanolamine)
• Sphingophospholipids (e.g., sphingomyelin)
• Glycolipids: Lipids with carbohydrate groups, mainly found in nerve tissue.
 • Cerebrosides
• Gangliosides
• Sulfatides
• Lipoproteins: Complexes of lipids and proteins for lipid transport in blood.
3. Derived Lipids
• Steroids: Compounds with a cyclopentanoperhydrophenanthrene ring system.
• Cholesterol: Key membrane component and precursor for steroid hormones, bile acids, and vitamin D.
• Fatty acids: Saturated and unsaturated, including essential fatty acids (linoleic, linolenic, arachidonic acids).
Properties of Lipids
• Hydrophobicity: Insoluble in water, soluble in organic solvents (e.g., ether, chloroform).
• Structural diversity: Range from simple fatty acids to complex molecules like phospholipids and steroids.
• Physical state: Varies with saturation; saturated fats are solid at room temperature, unsaturated fats are liquid.
• Isomerism: Unsaturated fatty acids exist as cis (natural) or trans (industrial) isomers, affecting melting point and
biological effects.
• Amphipathic nature: Some lipids (e.g., phospholipids) have both hydrophilic and hydrophobic regions, enabling
membrane formation.
Biological Functions of Lipids
• Structural: Phospholipids and cholesterol form the basis of biological membranes.
• Energy storage: Triglycerides are the main energy reserve, yielding more than twice the energy per gram as
carbohydrates or proteins.
 • Insulation and protection: Subcutaneous fat insulates the body; fat around organs provides mechanical protection.
• Regulatory: Steroid hormones (e.g., cortisol, sex hormones) and eicosanoids (prostaglandins, leukotrienes) regulate
numerous physiological processes.
• Metabolic: Cholesterol derivatives (bile acids, vitamin D) are essential for digestion and mineral metabolism.
• Transport: Lipoproteins enable the transport of hydrophobic lipids in the aqueous environment of blood.
• Lubrication and waterproofing: Waxes protect skin and hair, prevent dehydration.
Essential Fatty Acids
• Linoleic, linolenic, and arachidonic acids are essential for human health, as they cannot be synthesized by the body and
must be obtained from the diet.
• They are precursors for eicosanoids (prostaglandins, leukotrienes, thromboxanes), which have roles in inflammation,
immunity, and vascular function1.

102. Digestion, absorption of lipid digestion products. Factors required for digestion. The role of the bile salts.
Ans. Digestion of Lipids
1. Initial Digestion:
 • Mouth and Stomach: Lipid digestion begins minimally in the mouth and stomach through the action of lingual and
gastric lipases, which are more active in infants and on short/medium-chain triglycerides. However, their contribution in
adults is minor.
• Main Digestion in Small Intestine: The bulk of lipid digestion occurs in the small intestine.
2. Emulsification:
• Challenge: Dietary fats are hydrophobic and form large droplets, making them inaccessible to water-soluble digestive
enzymes.
• Solution: In the duodenum, bile salts (produced in the liver from cholesterol and stored in the gallbladder) are secreted
into the intestine. Bile salts act as detergents, reducing surface tension and breaking large fat droplets into
smaller emulsified droplets.
• Result: Emulsification increases the surface area for pancreatic enzymes to act1.
3. Enzymatic Hydrolysis:
• Pancreatic Lipase: The main enzyme, hydrolyzes triglycerides at positions 1 and 3, yielding 2-
monoacylglycerols and free fatty acids.
• Colipase: A cofactor secreted as a proenzyme and activated in the intestine; it binds to both the lipid and lipase,
facilitating enzyme action.
• Phospholipase A2: Hydrolyzes phospholipids to lysophospholipids and fatty acids (requires calcium ions).
• Cholesterol Esterase: Hydrolyzes dietary cholesterol esters to free cholesterol and fatty acids1.
Absorption of Lipid Digestion Products
1. Mixed Micelle Formation:
• Products: Long-chain fatty acids, 2-monoacylglycerols, cholesterol, lysophospholipids, and fat-soluble vitamins combine
with bile salts to form mixed micelles.
 • Structure: Hydrophobic core (lipids) and hydrophilic exterior (bile salts), allowing solubility in the aqueous intestinal
environment.
• Importance: Mixed micelles transport lipid digestion products to the brush border of enterocytes (intestinal cells)1.
2. Uptake by Enterocytes:
• Mechanism: Lipid components of micelles diffuse across the enterocyte membrane.
• Bile salts: Remain in the intestinal lumen and are reabsorbed in the ileum (enterohepatic circulation).
• Medium-chain fatty acids: Can be absorbed directly without micelles and enter the portal blood, bound to albumin.
3. Re-esterification and Chylomicron Formation:
• Inside Enterocytes: Long-chain fatty acids and monoacylglycerols are re-esterified to form triglycerides; cholesterol is re-
esterified; phospholipids are resynthesized.
• Chylomicrons: Newly synthesized triglycerides, cholesterol esters, and phospholipids are packaged with apolipoproteins
into chylomicrons, which are secreted into the lymphatic system and then enter the bloodstream1.
Factors Required for Lipid Digestion
• Bile Salts: For emulsification and micelle formation.
• Pancreatic Enzymes: Lipase, colipase, phospholipase A2, cholesterol esterase.
• Colipase: Activates pancreatic lipase.
• Calcium Ions: Required for phospholipase A2 activity.
• Peristalsis: Mechanical mixing aids emulsification.
The Role of Bile Salts
 • Emulsification: Bile salts are amphipathic molecules that reduce surface tension, breaking up fat into small droplets,
increasing the area for enzyme action.
• Micelle Formation: Essential for the solubilization of lipid digestion products, enabling their transport to the intestinal
mucosa for absorption.
• Absorption of Fat-Soluble Vitamins: Micelles also carry vitamins A, D, E, K for absorption.
• Enterohepatic Circulation: Most bile salts are reabsorbed in the ileum and recycled to the liver.
Without bile salts, fat digestion and absorption are severely impaired, leading to steatorrhea (fatty stools) and deficiencies
of fat-soluble vitamins1.

103. Resynthesis of lipids in the intestinal wall.

Ans. After digestion, dietary lipids are absorbed by the enterocytes (intestinal epithelial cells) of the small intestine, where they
undergo resynthesis (re-esterification) into complex lipids before entering the lymphatic and then the circulatory system.
1. Absorption of Lipid Digestion Products
• Products absorbed:
• Long-chain fatty acids
• 2-monoacylglycerols (from triglyceride digestion)
• Cholesterol
• Lysophospholipids
• Fat-soluble vitamins
• Mechanism:
These products are incorporated into mixed micelles with bile salts in the intestinal lumen, which transport them to the
 brush border of enterocytes. The lipid components diffuse into the enterocytes, while bile salts remain in the lumen to
be reabsorbed later1.
2. Resynthesis (Re-esterification) of Lipids in Enterocytes
Once inside the enterocyte, the lipid digestion products are reassembled into their complex forms:
A. Triacylglycerol (Triglyceride) Resynthesis
• Pathway:
1. Activation of fatty acids:
• Free long-chain fatty acids are activated to fatty acyl-CoA by acyl-CoA synthetase (requires ATP and CoA).
2. Esterification:
• 2-monoacylglycerol (from digestion) is acylated by fatty acyl-CoA to form diacylglycerol, then further
acylated to form triacylglycerol (TAG).
• This is known as the monoacylglycerol pathway, predominant in the intestine.
B. Cholesterol Esterification
• Cholesterol is esterified by acyl-CoA:cholesterol acyltransferase (ACAT) using fatty acyl-CoA to form cholesterol esters.
C. Phospholipid Resynthesis
• Lysophospholipids are reacylated to form phospholipids by acyltransferases.
3. Assembly and Secretion of Chylomicrons
• Formation:
 • Newly synthesized triacylglycerols, cholesterol esters, and phospholipids are packaged together with
apolipoproteins (mainly apoB-48) to form chylomicrons within the enterocyte’s endoplasmic reticulum and Golgi
apparatus.
• Secretion:
• Chylomicrons are exocytosed from the basolateral membrane of enterocytes into the lymphatic system (lacteals),
and then enter the bloodstream via the thoracic duct13.
4. Fate of Medium-Chain Fatty Acids
• Medium-chain fatty acids (from some dietary sources, e.g., milk) are absorbed directly into the portal blood without re-
esterification, bound to albumin, and transported to the liver1.

104. Deposition and mobilization of fats in adipose tissue: physiological significance, hormonal regulation. The role of insulin,
adrenaline, glucagon.
Ans. Physiological Significance
Adipose tissue serves as the body’s main energy reservoir, storing energy in the form of triacylglycerols (triglycerides). The
deposition (storage) and mobilization (breakdown) of fats are crucial for:
• Energy balance: Storing excess energy after meals and releasing it during fasting, exercise, or stress.
• Thermal insulation and protection: Fat cushions organs and insulates the body.
• Endocrine function: Adipose tissue secretes hormones (adipokines) that regulate metabolism.
Deposition (Storage) of Fats
Process:
• After a meal, dietary carbohydrates and fats are converted into fatty acids and glycerol in the liver and adipose tissue.
 • These are re-esterified to form triacylglycerols (TAGs) and stored in adipocytes.
Key steps:
• Uptake of glucose: Insulin stimulates glucose uptake into adipocytes, providing glycerol-3-phosphate for TAG synthesis.
• Uptake of fatty acids: Insulin also activates lipoprotein lipase (LPL) on capillary endothelium, releasing fatty acids from
circulating chylomicrons and VLDL for uptake by adipocytes.
• TAG synthesis: Fatty acids are esterified with glycerol-3-phosphate to form TAGs, which are stored in lipid droplets.
Mobilization (Breakdown) of Fats
Process:
• During fasting, exercise, or stress, stored TAGs are hydrolyzed to release free fatty acids (FFAs) and glycerol.
Key steps:
• Activation of hormone-sensitive lipase (HSL): This enzyme hydrolyzes TAGs to FFAs and glycerol.
• Release into blood: FFAs bind to albumin and are transported to tissues for oxidation; glycerol is sent to the liver for
gluconeogenesis.
Hormonal Regulation
Insulin (Anabolic Hormone)
• Promotes fat deposition:
• Stimulates glucose uptake (via GLUT4) and conversion to glycerol-3-phosphate.
• Activates lipoprotein lipase, increasing fatty acid uptake from circulating lipoproteins.
• Inhibits hormone-sensitive lipase, suppressing lipolysis.
• Stimulates fatty acid and TAG synthesis.
 • Net effect: Increases storage of TAGs in adipose tissue and decreases fat breakdown245.
Adrenaline (Epinephrine) and Glucagon (Catabolic Hormones)
• Promote fat mobilization:
• Bind to adrenergic and glucagon receptors on adipocytes, activating adenylate cyclase and increasing cAMP.
• cAMP activates protein kinase A (PKA), which phosphorylates and activates hormone-sensitive lipase.
• HSL hydrolyzes stored TAGs, releasing FFAs and glycerol.
• Adrenaline is especially important during acute stress and exercise; glucagon acts mainly during fasting245.
• Net effect: Increase lipolysis (fat breakdown) and release of FFAs for energy use by other tissues.

105. Transport forms of lipids, classification, composition. Apoproteins, types, role. Heterogeneity of lipid components of
lipoproteins. Atherogenicity.
Ans. Transport Forms of Lipids: Lipoproteins
Lipids are transported in the blood as lipoproteins-complex particles with a hydrophobic core (nonpolar lipids) and a
hydrophilic surface (phospholipids, free cholesterol, and apoproteins)1237.
Classification of Plasma Lipoproteins
Class Diameter Main Lipid Main Function
(nm) Content Apoproteins

Transport dietary
Triglycerides ApoB-48, ApoC, triglycerides from intestine
Chylomicrons 75–1200 (dietary) ApoE to tissues

Triglycerides ApoB-100, Transport liver-synthesized

VLDL 30–80 (endogenous) ApoC, ApoE triglycerides to tissues

ApoB-100, Transitional form between

IDL 25–50 Intermediate ApoE VLDL and LDL

Deliver cholesterol to
peripheral tissues ("bad
LDL 18–28 Cholesterol esters ApoB-100 cholesterol")

Reverse cholesterol
Phospholipids, ApoA-I, ApoA- transport to liver ("good
HDL 5–15 cholesterol II, ApoC cholesterol")

• Chylomicrons: Largest, lowest density, richest in triglycerides.

• VLDL: Rich in endogenous triglycerides.
• IDL: Transitional, not usually detected fasting.
• LDL: Rich in cholesterol esters, main carrier of cholesterol to tissues.
 • HDL: Rich in protein, involved in cholesterol removal from tissues.
Composition of Lipoproteins
• Core: Nonpolar lipids (triglycerides, cholesterol esters)
• Surface: Phospholipids, free cholesterol, apoproteins (apolipoproteins)
• Relative content: Chylomicrons and VLDL are triglyceride-rich; LDL is cholesterol-rich; HDL is protein- and phospholipid-
rich23.
Apolipoproteins (Apoproteins): Types and Roles
Apolipoproteins are the protein components of lipoproteins, essential for structure, receptor binding, and enzyme
activation137.

Apolipoprotein Main Lipoprotein(s) Function(s)

Activates LCAT (lecithin-cholesterol acyltransferase);

ApoA-I HDL cholesterol efflux from tissues

ApoA-II HDL Modulates HDL metabolism

ApoA-IV Chylomicrons, HDL Involved in chylomicron assembly

ApoB-48 Chylomicrons Structural protein, required for chylomicron formation

ApoB-100 VLDL, LDL Structural protein, binds LDL receptor

 Apolipoprotein Main Lipoprotein(s) Function(s)

ApoC-I VLDL, HDL Modulates lipoprotein metabolism

Chylomicrons, VLDL,
ApoC-II HDL Activates lipoprotein lipase (LPL)

Chylomicrons, VLDL,
ApoC-III HDL Inhibits LPL

Chylomicrons, VLDL,
ApoE HDL Mediates remnant uptake by liver (binds LDL receptor)

ApoD HDL Role in lipid metabolism (less well defined)

Key roles:
• Structural: ApoB-48 (chylomicrons), ApoB-100 (VLDL, LDL)
• Enzyme activation/inhibition: ApoC-II (activates LPL), ApoC-III (inhibits LPL)
• Receptor binding: ApoB-100 (LDL receptor), ApoE (remnant receptor)
• Cholesterol transport: ApoA-I (HDL, LCAT activation)
Heterogeneity of Lipid Components
• Lipid composition varies by class and function:
• Chylomicrons/VLDL: Triglyceride-rich, deliver fatty acids to tissues.
 • LDL: Cholesterol ester-rich, delivers cholesterol to cells.
• HDL: Phospholipid and protein-rich, collects excess cholesterol.
• Lipoprotein particles are heterogeneous within each class, with variations in size, density, and apoprotein composition,
affecting their metabolic fate and atherogenic potential3.
Atherogenicity
• LDL ("bad cholesterol"): High levels are strongly associated with atherosclerosis and cardiovascular disease. LDL delivers
cholesterol to artery walls, promoting plaque formation.
• HDL ("good cholesterol"): Involved in reverse cholesterol transport, removing cholesterol from tissues and atheromas,
and returning it to the liver for excretion. High HDL is protective.
• VLDL and remnants: Also considered atherogenic, especially when present in excess.
• Atherogenicity is influenced by:
• Particle size and density: Small, dense LDL is more atherogenic.
• Apolipoprotein content: ApoB-containing particles (chylomicron remnants, VLDL, LDL) are more atherogenic.
• Oxidative modification: Oxidized LDL is particularly damaging to vascular endothelium3.

106. Phospholipids, chemical structure, representatives, biological functions: structural role. Phospholipids as a source of
arachidonic acid for the biosynthesis of prostaglandins.
Ans. Chemical Structure of Phospholipids
Phospholipids are a major class of complex lipids and key components of biological membranes. Their general structure
includes:
• Backbone:
 • Glycerol (in glycerophospholipids) or sphingosine (in sphingophospholipids).
• Fatty Acids:
• Two fatty acid chains (usually at positions 1 and 2 of glycerol), which can be saturated or unsaturated.
• Phosphate Group:
• Attached to the third carbon of glycerol.
• Alcoholic (Polar) Head Group:
• Such as choline, ethanolamine, serine, or inositol, linked to the phosphate group.
Generalized structure of a glycerophospholipid:
Glycerol backbone + 2 fatty acids + phosphate + polar head group4.
Main Representatives
• Phosphatidylcholine (Lecithin):
• Contains choline as the head group; abundant in cell membranes and lipoproteins.
• Phosphatidylethanolamine (Cephalin):
• Contains ethanolamine; found in nervous tissue and cell membranes.
• Phosphatidylserine:
• Contains serine; important for membrane structure and signaling.
• Phosphatidylinositol:
• Contains inositol; involved in signal transduction.
• Sphingomyelin:
 • A sphingophospholipid with sphingosine backbone and choline head group; major component of myelin sheath in
nerve tissue47.
Biological Functions and Structural Role
• Structural Role in Membranes:
• Phospholipids are the primary building blocks of all biological membranes, forming a bilayer in which hydrophobic
fatty acid tails face inward and hydrophilic head groups face outward.
• They provide membrane fluidity, flexibility, and selective permeability.
• The specific composition of phospholipids influences membrane properties and the function of embedded
proteins47.
• Other Functions:
• Reservoir for signaling molecules: Phospholipids serve as precursors for second messengers (e.g.,
phosphatidylinositol bisphosphate for IP₃ and DAG).
• Lipoprotein structure: They form the surface monolayer of lipoproteins, essential for lipid transport in blood.
• Cell recognition and signaling: Some phospholipids are involved in cell signaling and apoptosis (e.g.,
phosphatidylserine externalization during apoptosis).
Phospholipids as a Source of Arachidonic Acid for Prostaglandin Biosynthesis
• Arachidonic acid (20:4, ω-6) is a polyunsaturated fatty acid commonly esterified at the second (sn-2) position of
membrane phospholipids, especially phosphatidylcholine and phosphatidylethanolamine45.
• Release of Arachidonic Acid:
• Upon stimulation (e.g., by hormones, cytokines, or injury), phospholipase A₂ hydrolyzes the sn-2 bond of
phospholipids, releasing free arachidonic acid into the cytosol.
 • Biosynthesis of Prostaglandins:
• Free arachidonic acid is the substrate for cyclooxygenase (COX) and lipoxygenase enzymes, leading to the
production of eicosanoids: prostaglandins, thromboxanes, and leukotrienes.
• Prostaglandins are potent local hormones (autacoids) involved in inflammation, pain, fever, vascular tone, and
many other physiological processes5.

107. Membrane proteins and lipids: structural and functional heterogeneity. Properties and functions of membranes.
Ans. Structural and Functional Heterogeneity of Membrane Proteins and Lipids
Membrane Lipids
• Diversity of Lipid Types: Biological membranes are primarily composed of a lipid bilayer containing various lipids:
• Phospholipids: The most abundant, including glycerophospholipids (e.g., phosphatidylcholine,
phosphatidylethanolamine, phosphatidylserine) and sphingophospholipids (e.g., sphingomyelin)1.
• Glycolipids: Such as cerebrosides and gangliosides, especially abundant in nervous tissue and involved in cell
recognition13.
• Cholesterol: Modulates membrane fluidity and stability, and is more abundant in animal cell membranes16.
• Heterogeneity and Asymmetry:
• Different membranes (plasma membrane, mitochondrial, ER, myelin, etc.) have distinct lipid compositions and
ratios, reflecting their specialized functions36.
• The two leaflets of the bilayer are asymmetric: for example, phosphatidylserine and phosphatidylethanolamine
are more common on the cytoplasmic side, while phosphatidylcholine and sphingomyelin are more abundant on
the extracellular side1.
 • The presence of unsaturated fatty acids (cis configuration) in phospholipids introduces kinks, increasing
membrane fluidity and affecting protein function1.
Membrane Proteins
• Types of Membrane Proteins:
• Integral (transmembrane) proteins: Span the lipid bilayer, often with multiple membrane-spanning domains.
They function as channels, transporters, receptors, and enzymes.
• Peripheral proteins: Loosely attached to the membrane surface, often via interactions with integral proteins or
lipid head groups. They are involved in signaling, structural support, and enzymatic activity4.
• Lipid-anchored proteins: Covalently attached to lipids within the bilayer.
• Heterogeneity:
• Each membrane has a unique set of proteins, reflecting the specific functions of the cell or organelle.
• Proteins can form complexes, be glycosylated (glycoproteins), or interact with cytoskeletal elements, adding
further functional diversity4.
• Membrane proteins differ in structure (alpha-helical, beta-barrel), size, and the number of subunits, contributing
to the diversity of membrane functions24.
Properties of Biological Membranes
• Fluid Mosaic Model: Membranes are dynamic, with lipids and proteins able to move laterally within the layer, giving
flexibility and allowing for membrane remodeling and the formation of microdomains (rafts)16.
• Selective Permeability: The hydrophobic core restricts passage of polar and charged molecules, while allowing small
nonpolar molecules to diffuse. Specific proteins mediate the transport of ions, nutrients, and waste16.
• Asymmetry: Lipid and protein composition differs between the inner and outer leaflets, crucial for functions such as cell
signaling and apoptosis1.
 • Self-Sealing: Membranes can reseal after minor damage, essential for vesicle trafficking and cell integrity.
• Electrical Properties: The membrane can maintain ion gradients and membrane potential, vital for nerve impulse
transmission and muscle contraction3.
Functions of Biological Membranes
• Compartmentalization: Separate the cell from its environment and create distinct organelles within eukaryotic cells,
allowing specialized functions16.
• Transport: Regulate movement of substances via passive diffusion, facilitated diffusion, active transport, endocytosis,
and exocytosis.
• Signal Transduction: Membrane proteins act as receptors for hormones, neurotransmitters, and other signals, initiating
intracellular signaling cascades4.
• Cell Recognition and Communication: Glycoproteins and glycolipids are involved in immune recognition, tissue
organization, and intercellular communication4.
• Structural Support: Membranes anchor the cytoskeleton and extracellular matrix, maintaining cell shape and enabling
movement.
• Energy Conversion: Mitochondrial and chloroplast membranes house the machinery for ATP synthesis and
photosynthesis, respectively6.
• Enzymatic Activity: Many membrane proteins function as enzymes, catalyzing key metabolic reactions at the membrane
surface4.

108. Connection of fatty acid oxidation with the tricarboxylic acid cycle and the respiratory chain. Biological role.
Ans. Connection of Fatty Acid Oxidation with the Tricarboxylic Acid Cycle and the Respiratory Chain: Biological Role
1. Overview: Fatty Acid Oxidation (β-Oxidation)
 • Location: Mitochondrial matrix of liver, heart, muscle, kidney, and other tissues13.
• Process: Fatty acids are activated to acyl-CoA and transported into mitochondria (via the carnitine shuttle), where they
undergo β-oxidation-a cyclic sequence that shortens the fatty acid by two carbons per turn, generating acetyl-CoA,
NADH, and FADH₂1.
2. Connection to the Tricarboxylic Acid (TCA) Cycle
• Acetyl-CoA Production: Each cycle of β-oxidation produces one acetyl-CoA1.
• Entry into TCA Cycle: Acetyl-CoA condenses with oxaloacetate to form citrate, entering the TCA cycle12.
• TCA Cycle Role: The acetyl group is fully oxidized to CO₂, generating additional NADH and FADH₂2.
3. Connection to the Respiratory Chain (Electron Transport Chain, ETC)
• NADH and FADH₂ Production: Both β-oxidation and the TCA cycle produce NADH and FADH₂13.
• Electron Donation: NADH and FADH₂ donate electrons to the ETC (located in the inner mitochondrial membrane)3.
• NADH: Enters at Complex I, yielding ~2.5–3 ATP per NADH.
• FADH₂: Enters at Complex II, yielding ~1.5–2 ATP per FADH₂3.
• Oxygen as Final Acceptor: Electrons are ultimately transferred to oxygen, forming water.
• ATP Synthesis: The energy released during electron transfer is used to pump protons, creating a proton gradient that
drives ATP synthesis via ATP synthase (oxidative phosphorylation)3.
4. Integrated Pathway Summary
Fatty acid → β-oxidation → Acetyl-CoA + NADH + FADH₂ →
• Acetyl-CoA → TCA cycle → CO₂ + NADH + FADH₂
• NADH/FADH₂ → ETC → ATP (energy) + H₂O
5. Biological Role
• Major Energy Source: Fatty acid oxidation is the primary energy source during fasting, prolonged exercise, and between
meals, especially in heart, liver, and skeletal muscle1.
• High ATP Yield: Complete oxidation of one palmitic acid (16C) yields 106–131 ATP molecules, far more than glucose1.
• Metabolic Integration: Links lipid, carbohydrate, and protein metabolism; acetyl-CoA from fatty acids can also be used
for ketone body synthesis during prolonged fasting or diabetes.
• Homeostasis: Provides energy when glucose is scarce, spares glucose for tissues like the brain and red blood cells, and
prevents hypoglycemia1.
6. Key Regulatory Points
• Supply of Fatty Acids: Controlled by hormone-sensitive lipase in adipose tissue (regulated by insulin, glucagon,
adrenaline)1.
• Entry into Mitochondria: Carnitine palmitoyltransferase I (CPT I) is inhibited by malonyl-CoA, preventing simultaneous
fatty acid synthesis and oxidation1.
• TCA Cycle Activity: Regulated by energy status (ATP/ADP, NADH/NAD⁺ ratios)2.

109. Oxidation of higher fatty acids. Localization of the process. The sequence of reactions.
Ans. Oxidation of Higher Fatty Acids: Localization and Sequence of Reactions
Localization of Fatty Acid Oxidation
• Primary Site: The oxidation of higher (long-chain) fatty acids occurs mainly in the mitochondrial matrix of cells.
• Tissues: This process is especially active in the liver, heart, skeletal muscle, kidney, brain, lungs, testes, and adipose
tissue13.
 • Prerequisite: Fatty acids must first be activated and transported into mitochondria before oxidation can proceed.
Sequence of Reactions: β-Oxidation Pathway
The main pathway for the oxidation of higher fatty acids is β-oxidation, which systematically removes two-carbon units as
acetyl-CoA from the carboxyl end of the fatty acid chain.
1. Activation of Fatty Acids (Cytosol)
• Enzyme: Acyl-CoA synthetase (thiokinase)
• Reaction: Fatty acid + CoA + ATP → Acyl-CoA + AMP + PPi
• Location: Outer mitochondrial membrane or cytosol
• Purpose: Converts free fatty acids to their active form (acyl-CoA), which can be metabolized1.
2. Transport into Mitochondria (Carnitine Shuttle)
• Long-chain acyl-CoA cannot cross the inner mitochondrial membrane directly.
• Steps:
• Carnitine acyltransferase I (outer membrane): Transfers acyl group from CoA to carnitine, forming acyl-carnitine.
• Translocase: Transports acyl-carnitine across the inner membrane.
• Carnitine acyltransferase II (inner membrane): Transfers acyl group back to CoA, regenerating acyl-CoA in the
matrix; carnitine returns to the cytosol1.
3. β-Oxidation Spiral (Mitochondrial Matrix)
Each cycle shortens the fatty acyl-CoA by two carbons, releasing one acetyl-CoA, one NADH, and one FADH₂. The cycle repeats
until the fatty acid is completely degraded.
The four steps of each β-oxidation cycle:
 1. Dehydrogenation (Oxidation)
• Enzyme: Acyl-CoA dehydrogenase
• Reaction: Acyl-CoA → trans-Δ²-enoyl-CoA
• Cofactor: FAD (produces FADH₂)
• Purpose: Introduces a double bond between the α and β carbons1.
2. Hydration
• Enzyme: Enoyl-CoA hydratase
• Reaction: trans-Δ²-enoyl-CoA + H₂O → L-β-hydroxyacyl-CoA
• Purpose: Adds water across the double bond, forming a hydroxyl group on the β-carbon1.
3. Second Dehydrogenation (Oxidation)
• Enzyme: β-hydroxyacyl-CoA dehydrogenase
• Reaction: L-β-hydroxyacyl-CoA → β-ketoacyl-CoA
• Cofactor: NAD⁺ (produces NADH)
• Purpose: Oxidizes the hydroxyl group to a keto group1.
4. Thiolytic Cleavage
• Enzyme: β-ketothiolase
• Reaction: β-ketoacyl-CoA + CoA → Acetyl-CoA + (Fatty acyl-CoA shortened by 2C)
• Purpose: Cleaves the β-ketoacyl-CoA, releasing acetyl-CoA and a new acyl-CoA (two carbons shorter)1.
The process repeats until the entire fatty acid is converted to acetyl-CoA units.
Overall Reaction Example: Palmitic Acid (C16)
For palmitoyl-CoA (16 carbons):
• 7 cycles of β-oxidation produce:
• 8 acetyl-CoA
• 7 NADH
• 7 FADH₂
• 7 H₂O
• 7 H⁺
• Each acetyl-CoA enters the TCA cycle for further oxidation and ATP production1.

110. Biosynthesis of triacylglycerols in the liver and adipose tissue. Dependence of the biosynthesis rate on the nutritional
rhythm and food composition.
Ans. Pathway of Triacylglycerol (TAG) Biosynthesis
1. Sites of Synthesis
• Liver: Major site for TAG synthesis, especially after carbohydrate-rich meals. TAGs synthesized here are packaged into
VLDL for export to other tissues.
• Adipose Tissue: Main storage site for TAGs. Synthesis increases after meals for energy storage.
2. Precursors and Key Steps
• Glycerol-3-phosphate: Backbone for TAG synthesis.
 • Liver: Can be formed from glycerol (via glycerol kinase) or from glucose (via glycolysis: dihydroxyacetone
phosphate → glycerol-3-phosphate).
• Adipose tissue: Lacks glycerol kinase, so relies on glucose uptake (stimulated by insulin) and glycolysis for glycerol-
3-phosphate production.
• Fatty acyl-CoA: Activated form of fatty acids, produced by acyl-CoA synthetase.
3. Sequence of Reactions
1. Glycerol-3-phosphate + Fatty acyl-CoA → Lysophosphatidic acid (by acyltransferase)
2. Lysophosphatidic acid + Fatty acyl-CoA → Phosphatidic acid (by acyltransferase)
3. Phosphatidic acid → Diacylglycerol (DAG) (by phosphatidic acid phosphatase)
4. DAG + Fatty acyl-CoA → Triacylglycerol (TAG) (by diacylglycerol acyltransferase)
• In summary:
Glycerol-3-phosphate + 3 Fatty acyl-CoA → Triacylglycerol + 3 CoA
4. Fate of Synthesized TAGs
• Liver: TAGs are packaged into VLDL and secreted into the blood for transport to adipose and other tissues.
• Adipose tissue: TAGs are stored in lipid droplets for future energy needs.
Dependence of TAG Biosynthesis Rate on Nutritional Rhythm and Food Composition
Nutritional Rhythm
• Fed State (Postprandial):
• High carbohydrate intake: Increases glycolysis, raising glycerol-3-phosphate and acetyl-CoA levels, boosting fatty
acid and TAG synthesis.
 • Insulin secretion: Stimulates glucose uptake (adipose), activates lipogenic enzymes, and increases TAG synthesis
and storage.
• Fasting State:
• Low insulin, high glucagon: TAG synthesis decreases, and lipolysis (breakdown) predominates in adipose tissue.
Food Composition
• Carbohydrate-rich Diet:
• Provides glucose for both glycerol-3-phosphate and acetyl-CoA (for fatty acid synthesis), strongly stimulating TAG
synthesis in both liver and adipose tissue.
• Fat-rich Diet:
• Increases supply of fatty acids for TAG synthesis, but if carbohydrates are low, glycerol-3-phosphate availability
(especially in adipose tissue) can limit TAG formation.
• Protein-rich Diet:
• Amino acids can be converted to glucose (gluconeogenesis), indirectly supporting TAG synthesis if carbohydrate
intake is low.
Hormonal Regulation
• Insulin: Major stimulator of TAG synthesis.
• Increases glucose uptake (via GLUT4 in adipose tissue), upregulates lipogenic and glycolytic enzymes, and
promotes fatty acid and TAG synthesis.
• Glucagon and Catecholamines: Inhibit TAG synthesis and stimulate lipolysis during fasting or stress.
111. The relationship of lipid and carbohydrate metabolism. Diagram of the conversion of glucose into fats. The role of the
pentose phosphate pathway of glucose metabolism in lipid synthesis.
Ans. Lipid and carbohydrate metabolism are tightly interconnected. Carbohydrates can be converted into lipids when energy
intake exceeds immediate needs, and lipids can be oxidized to provide energy when carbohydrates are scarce. This metabolic
flexibility is crucial for energy storage, mobilization, and biosynthesis.
Conversion of Glucose into Fats (Lipogenesis)
Pathway Diagram and Steps
1. Glucose Uptake and Glycolysis
• Glucose enters cells and is metabolized via glycolysis to produce pyruvate.
2. Pyruvate to Acetyl-CoA
• Pyruvate enters mitochondria and is converted to acetyl-CoA by the pyruvate dehydrogenase complex.
3. Citrate Shuttle
• Acetyl-CoA combines with oxaloacetate to form citrate, which is transported out of the mitochondria into the
cytosol.
4. Cytosolic Acetyl-CoA Formation
• In the cytosol, citrate is cleaved by ATP-citrate lyase to regenerate acetyl-CoA and oxaloacetate.
5. Fatty Acid Synthesis
• Acetyl-CoA is converted to malonyl-CoA by acetyl-CoA carboxylase (rate-limiting step, requires biotin and ATP).
• Fatty acid synthase complex elongates the chain, using acetyl-CoA and malonyl-CoA, with NADPH as a reducing
agent, to form palmitate (C16:0).
6. Triglyceride (TAG) Synthesis
 • Fatty acids are esterified with glycerol-3-phosphate (derived from glycolysis) to form triglycerides, which are
stored in adipose tissue or exported as VLDL from the liver.
Summary Diagram:
text
Glucose
↓ (glycolysis)
Pyruvate
↓ (pyruvate dehydrogenase)
Acetyl-CoA (mitochondria)
↓ + Oxaloacetate → Citrate (TCA cycle)
Citrate (transported to cytosol)
↓ (ATP-citrate lyase)
Acetyl-CoA (cytosol)
↓ (acetyl-CoA carboxylase)
Malonyl-CoA
↓ (fatty acid synthase, NADPH required)
Fatty acids
↓ (esterification with glycerol-3-phosphate)
Triacylglycerols (fats)
The Role of the Pentose Phosphate Pathway (PPP) in Lipid Synthesis
 • NADPH Production:
The PPP (hexose monophosphate shunt) oxidizes glucose-6-phosphate to ribulose-5-phosphate, generating NADPH in
the process. NADPH is essential for reductive biosynthesis, especially for fatty acid and cholesterol synthesis in the
cytosol6.
• Biosynthetic Support:
• Fatty acid synthesis: Each cycle of fatty acid chain elongation by fatty acid synthase requires NADPH.
• Cholesterol synthesis: Multiple steps in the pathway require NADPH as a reducing agent36.
• Ribose-5-phosphate Production:
The PPP also provides ribose-5-phosphate for nucleotide synthesis, supporting cell growth and division.
Integration and Regulation
• Fed State:
• High glucose and insulin levels promote glycolysis, PPP (for NADPH), and lipogenesis.
• Insulin stimulates acetyl-CoA carboxylase and fatty acid synthase, and increases glucose uptake into adipose
tissue.
• Fasting State:
• Lipolysis and fatty acid oxidation are favored; lipogenesis is suppressed.
Key Points
• Glucose can be converted into fatty acids and triglycerides when energy intake exceeds immediate needs.
• The pentose phosphate pathway is critical for supplying NADPH for fatty acid and cholesterol biosynthesis.
• Lipogenesis is hormonally regulated, mainly stimulated by insulin and suppressed by glucagon and adrenaline23.
112. Nutritional and endogenous cholesterol, structure, biological role. Cholesterol biosynthesis: stages, process regulation,
excretion of cholesterol from the body.
Ans. Nutritional and Endogenous Cholesterol
Sources
• Nutritional (Exogenous) Cholesterol:
• Obtained from animal-based foods (e.g., egg yolk, meat, dairy, shellfish).
• Absorbed in the intestine, incorporated into chylomicrons, and transported via lymph and blood to tissues and the
liver21.
• Endogenous Cholesterol:
• Synthesized de novo in the body, primarily in the liver (major site), but also in the intestine, adrenal cortex, and
reproductive organs.
• Endogenous synthesis accounts for the majority of cholesterol in the body1.
Structure of Cholesterol
• Steroid Structure:
• Cholesterol is a sterol (steroid alcohol) with a characteristic structure: four fused hydrocarbon rings
(cyclopentanoperhydrophenanthrene nucleus), a hydrocarbon tail, a hydroxyl group at C3, and a double bond
between C5 and C6.
• Amphipathic Nature:
• The hydroxyl group is polar (hydrophilic), while the rest of the molecule is nonpolar (hydrophobic)21.
Biological Role of Cholesterol
1. Structural Role:
 • Essential component of all animal cell membranes, modulating fluidity, permeability, and membrane protein
function12.
2. Precursor for Biosynthesis:
• Bile acids: Required for fat digestion and absorption.
• Steroid hormones: Precursor for adrenal corticosteroids, sex hormones (testosterone, estrogen, progesterone).
• Vitamin D: 7-dehydrocholesterol in the skin is converted to vitamin D3 upon UV exposure.
• Other molecules: Ubiquinone (coenzyme Q), dolichol (glycoprotein synthesis)1.
3. Lipoprotein Component:
• Present as free cholesterol and cholesterol esters in plasma lipoproteins, involved in lipid transport.
Cholesterol Biosynthesis: Stages
Location: Cytosol and endoplasmic reticulum, mainly in the liver.
Main Stages:
1. Synthesis of Mevalonate from Acetyl-CoA
• Acetyl-CoA → Acetoacetyl-CoA → HMG-CoA
• HMG-CoA → Mevalonate (via HMG-CoA reductase, the rate-limiting and regulatory step)1.
2. Formation of Activated Isoprenoid Units
• Mevalonate → Mevalonate-5-phosphate → Mevalonate-5-pyrophosphate → Isopentenyl pyrophosphate (IPP)
• IPP isomerizes to dimethylallyl pyrophosphate (DMAPP).
3. Synthesis of Squalene
• Condensation of six isoprene units (5C each) → Squalene (30C)
 4. Cyclization to Lanosterol and Conversion to Cholesterol
• Squalene → Squalene epoxide → Lanosterol → Cholesterol
• Multiple steps involving demethylation and isomerization1.
Regulation of Cholesterol Biosynthesis
• HMG-CoA Reductase:
• The key regulatory enzyme, subject to:
• Feedback inhibition by cholesterol (end-product).
• Hormonal regulation: Insulin and thyroid hormones stimulate, while glucagon and glucocorticoids inhibit its
activity.
• Phosphorylation: AMP-activated protein kinase inactivates HMG-CoA reductase during low energy states
(high AMP).
• Gene expression: Regulated by sterol regulatory element-binding proteins (SREBPs)1.
• Dietary Influence:
• High dietary cholesterol suppresses endogenous synthesis; low intake increases synthesis.
Excretion of Cholesterol from the Body
• Bile Acids:
• The major route. Cholesterol is converted to bile acids in the liver, secreted into bile, and excreted in feces. Some
bile acids are reabsorbed (enterohepatic circulation), but a portion is lost daily.
• Neutral Sterols:
• Unmodified cholesterol is also excreted in bile and eliminated in feces.
 • Minor Routes:
• Small amounts are converted to steroid hormones or vitamin D and excreted as their metabolites12.

113. The role of lipoproteins in cholesterol transport. Hypercholesterolemia is a risk factor for atherosclerosis. Biochemistry of
gallstone disease.
Ans. Lipoproteins and Cholesterol Transport
Cholesterol, being insoluble in water, is transported in the blood as part of lipoprotein complexes. These complexes have a
hydrophobic core (cholesteryl esters and triglycerides) and a hydrophilic surface (phospholipids, free cholesterol, and
apoproteins)23.
Major Classes of Plasma Lipoproteins and Their Roles:

Lipoprotein Main Apoproteins Function

Cholesterol
Role

ApoB-48, ApoC, Transport dietary cholesterol and

Chylomicrons Minor ApoE triglycerides from intestine to tissues

ApoB-100, Carry endogenous triglycerides and some

VLDL Some ApoC, ApoE cholesterol from liver to tissues

Main carrier of cholesterol to peripheral

LDL Major ApoB-100 tissues (“bad cholesterol”)
 Lipoprotein Main Apoproteins Function
Cholesterol
Role

Collects excess cholesterol from tissues

and returns it to the liver (“good
HDL Major ApoA-I, ApoA-II cholesterol”)

• LDL delivers cholesterol to cells via receptor-mediated endocytosis. Excess LDL cholesterol can deposit in arterial walls,
contributing to atherosclerosis.
• HDL is involved in reverse cholesterol transport, removing excess cholesterol from tissues and atheromas and delivering
it to the liver for excretion or recycling23.
Hypercholesterolemia as a Risk Factor for Atherosclerosis
Hypercholesterolemia is defined as an abnormally high level of cholesterol in the blood, particularly elevated LDL cholesterol3.
• Atherosclerosis is a chronic disease characterized by the accumulation of cholesterol-rich plaques in arterial walls,
leading to narrowing, reduced elasticity, and risk of thrombosis.
• Risk factors: Elevated LDL, low HDL, smoking, hypertension, diabetes, family history, and age all increase the risk.
• Pathogenesis: LDL infiltrates the arterial intima, becomes oxidized, and is taken up by macrophages, forming foam cells
and fatty streaks. Over time, this leads to plaque formation, arterial narrowing, and potential for rupture and
thrombosis3.
• Clinical impact: Atherosclerosis underlies most cases of coronary artery disease, myocardial infarction, and ischemic
stroke.
Biochemistry of Gallstone Disease
Gallstone disease (cholelithiasis) often involves cholesterol stones, which form when bile becomes supersaturated with
cholesterol13:
• Cholesterol in bile: Cholesterol is secreted into bile by the liver, usually solubilized by bile acids and phospholipids.
• Stone formation: If the ratio of cholesterol to bile acids and phospholipids is too high, cholesterol precipitates and forms
crystals, which can aggregate into stones.
• Contributing factors:
• High cholesterol secretion (hypercholesterolemia, obesity, high-fat diet)
• Low bile acid synthesis or secretion (liver disease, rapid weight loss)
• Impaired gallbladder motility (stasis)
• Clinical consequences: Gallstones can obstruct the bile ducts, causing pain, inflammation (cholecystitis), jaundice, and
pancreatitis.

114. Biosynthesis of higher fatty acids, process localization, preparatory stage, sequence of reactions.
Ans. Localization of Fatty Acid Biosynthesis
• Cellular Location: The biosynthesis of higher (long-chain) fatty acids occurs in the cytosol of cells.
• Main Tissues:
• Liver (primary site in humans)
• Adipose tissue
• Mammary glands (during lactation)
• Kidney, brain (to a lesser extent)
Preparatory Stage: Generation of Precursors and Reducing Power
1. Source of Acetyl-CoA:
• Acetyl-CoA is generated in mitochondria from the oxidation of carbohydrates (via glycolysis and pyruvate
dehydrogenase), fatty acids, and some amino acids.
• Citrate Shuttle: Since the mitochondrial membrane is impermeable to acetyl-CoA, it is first converted to citrate
(by condensation with oxaloacetate), transported to the cytosol, and then cleaved by ATP-citrate lyase to
regenerate acetyl-CoA and oxaloacetate in the cytosol1.
2. Source of NADPH:
• NADPH is required as the reducing agent for fatty acid synthesis.
• Main sources: Pentose phosphate pathway and the malic enzyme (malate → pyruvate + NADPH)12.
Sequence of Reactions in Fatty Acid Biosynthesis
1. Carboxylation of Acetyl-CoA to Malonyl-CoA (Committed, Regulatory Step)
• Enzyme: Acetyl-CoA carboxylase (ACC)
• Cofactor: Biotin
• Reaction:
Acetyl-CoA+CO2+ATP→Malonyl-CoA+ADP+PiAcetyl-CoA+CO2+ATP→Malonyl-CoA+ADP+Pi
• Regulation: ACC is activated by citrate and insulin, and inhibited by long-chain acyl-CoA, glucagon, and
phosphorylation1.
2. Fatty Acid Synthase (FAS) Complex Reactions
• Type: Multienzyme complex with multiple functional domains, including an acyl carrier protein (ACP).
 • Main Steps:
1. Priming: Acetyl group from acetyl-CoA is transferred to the FAS complex (to the ACP and then to a cysteine
residue).
2. Loading: Malonyl group from malonyl-CoA is transferred to ACP.
3. Condensation: Acetyl group (on cysteine) and malonyl group (on ACP) condense, releasing CO₂ and forming a 4-
carbon β-ketoacyl-ACP.
4. Reduction: β-keto group is reduced to a hydroxyl group (using NADPH).
5. Dehydration: Removal of water forms a trans double bond.
6. Second Reduction: Double bond is reduced to a saturated acyl group (using NADPH).
7. Translocation: The growing acyl chain is transferred back to the cysteine residue, and a new malonyl group is
loaded onto ACP.
8. Cycle Repeats: Steps 3–7 repeat, each time extending the chain by two carbons, until a 16-carbon saturated fatty
acid (palmitate) is formed1.
• Termination: After seven cycles, the 16-carbon palmitoyl group is hydrolyzed from the enzyme complex,
releasing palmitic acid (palmitate) as the primary product.
3. Further Modifications (Elongation and Desaturation)
• Elongation: Palmitate can be further elongated in the endoplasmic reticulum by elongase enzymes using malonyl-CoA.
• Desaturation: Introduction of double bonds by desaturase enzymes (requires O₂, NADH, and cytochrome b₅)1.

115. Ketogenesis, process localization, use of ketone bodies as energy precursors. Ketonemia and ketonuria. Causes of ketosis
Methods for the determination of ketone bodies in urine.
Ans. Ketogenesis: Process and Localization
• Definition:
Ketogenesis is the metabolic process by which the liver converts excess acetyl-CoA, primarily from fatty acid β-oxidation,
into ketone bodies: acetoacetate, β-hydroxybutyrate, and acetone.
• Site:
Mitochondria of hepatocytes (liver cells) are the exclusive site of significant ketone body synthesis1.
• Sequence of Reactions:
1. Condensation of two acetyl-CoA to form acetoacetyl-CoA (enzyme: thiolase).
2. Acetoacetyl-CoA + acetyl-CoA → HMG-CoA (enzyme: HMG-CoA synthase).
3. HMG-CoA is cleaved to acetoacetate and acetyl-CoA (enzyme: HMG-CoA lyase).
4. Acetoacetate is:
• Reduced to β-hydroxybutyrate (enzyme: β-hydroxybutyrate dehydrogenase, using NADH).
• Decarboxylated spontaneously to acetone (non-enzymatic).
Use of Ketone Bodies as Energy Precursors
• Export and Utilization:
Ketone bodies are released from the liver into the blood and transported to extrahepatic tissues (muscle, heart, kidney,
brain during prolonged fasting), where they are converted back to acetyl-CoA and oxidized in the TCA cycle for ATP
production1.
• Significance:
• Spares glucose: During fasting, starvation, or diabetes, ketone bodies provide an alternative fuel, especially for
the brain.
 • Efficient energy source: Heart and skeletal muscle preferentially use ketone bodies when available.
Ketonemia, Ketonuria, and Causes of Ketosis
• Ketonemia:
Elevated levels of ketone bodies in the blood. Normal: <1 mg/100 mL; ketosis: much higher levels1.
• Ketonuria:
Excretion of ketone bodies in urine, occurs when blood levels exceed the renal threshold.
• Ketosis:
A metabolic state characterized by accumulation of ketone bodies in blood and tissues, with increased urinary excretion.
Causes of Ketosis
1. Starvation or prolonged fasting:
Depletion of carbohydrate reserves increases lipolysis and fatty acid oxidation.
2. Uncontrolled diabetes mellitus:
Insulin deficiency leads to enhanced lipolysis and massive hepatic ketogenesis.
3. High-fat, low-carbohydrate diets:
Promote fatty acid oxidation and ketone body formation.
4. Severe exercise, postabsorptive state, or certain hormonal imbalances:
Increased demand for alternative fuels.
5. Prolonged anesthesia or certain metabolic disorders.
Methods for Determination of Ketone Bodies in Urine
• Qualitative Tests:
 • Rothera’s Test:
Detects acetoacetate and acetone using sodium nitroprusside and ammonium sulfate; a purple ring indicates a
positive result.
• Gerhardt’s Test:
Uses ferric chloride to detect acetoacetic acid; a burgundy color indicates presence.
• Acetest Tablets:
Commercially available tablets containing sodium nitroprusside; color change indicates presence of ketone
bodies.
• Quantitative Methods:
• Spectrophotometric assays or enzymatic methods can measure concentrations of specific ketone bodies in urine
or blood.

116. Indicators characterizing the state of lipid metabolism in the body. Content and methods of determination in blood.
Ans. Key Indicators in Blood
To assess lipid metabolism, several laboratory indicators are routinely measured in blood. These include:
• Total cholesterol
• Triglycerides (TG)
• High-density lipoprotein cholesterol (HDL-C)
• Low-density lipoprotein cholesterol (LDL-C)
• Very-low-density lipoprotein cholesterol (VLDL-C)
• Free fatty acids (FFA)
 • Phospholipids
• Lipoprotein(a) [Lp(a)]
• Apolipoproteins (ApoA-I, ApoB, etc.)
• Ketone bodies (in certain metabolic states)
• Chylomicrons (postprandial or in lipid metabolism disorders)
These indicators provide information on the balance between lipid intake, synthesis, transport, utilization, and excretion, as
well as the risk for atherosclerosis and other metabolic diseases1.
Content (Reference Ranges) in Blood
• Total cholesterol: 3.9–6.5 mmol/L
• Triglycerides: 0.55–1.65 mmol/L (fasting)
• HDL-C: >1.0 mmol/L (men), >1.2 mmol/L (women)
• LDL-C: <3.0 mmol/L (optimal)
• VLDL-C: 0.2–1.7 mmol/L (calculated or measured)
• Free fatty acids: 0.2–0.8 mmol/L (fasting)
• Phospholipids: 2.5–3.5 mmol/L
• Lipoprotein(a): <0.3 g/L (varies by lab)
• Apolipoprotein A-I: 1.0–1.8 g/L
• Apolipoprotein B: 0.6–1.3 g/L
Reference ranges may vary by laboratory and population.
Methods of Determination
1. Enzymatic Colorimetric Methods
• Total cholesterol: Cholesterol esterase and oxidase convert cholesterol to cholestenone and hydrogen peroxide, which
reacts with a chromogen to produce a measurable color.
• Triglycerides: Lipase hydrolyzes triglycerides to glycerol, which is measured after further enzymatic reactions.
• HDL-C and LDL-C: After precipitation or selective inhibition of other lipoproteins, HDL or LDL cholesterol is measured
enzymatically.
• VLDL-C: Usually calculated (TG/2.2 in mmol/L) or measured by ultracentrifugation.
2. Electrophoresis and Ultracentrifugation
• Used for detailed lipoprotein fractionation and research purposes.
3. Immunoturbidimetric and Nephelometric Methods
• For apolipoproteins and Lp(a); based on antigen-antibody reactions.
4. Free Fatty Acids
• Enzymatic methods using acyl-CoA synthetase and acyl-CoA oxidase, producing hydrogen peroxide measured
colorimetrically.
5. Phospholipids
• Measured by enzymatic hydrolysis to release choline, which is then quantified.
6. Ketone Bodies
• Nitroprusside reaction (Rothera’s test) for acetoacetate and acetone; specific enzymatic or spectrophotometric methods
for β-hydroxybutyrate.
Clinical and Diagnostic Value
 • Hyperlipidemia (elevated cholesterol, LDL, TG): Indicates increased risk for atherosclerosis, coronary artery disease, and
pancreatitis.
• Low HDL: Associated with higher cardiovascular risk.
• High free fatty acids: Seen in uncontrolled diabetes, fasting, or stress.
• Elevated ketone bodies: Marker of ketosis/diabetic ketoacidosis.
• Apolipoprotein levels: ApoB (LDL marker) and ApoA-I (HDL marker) provide additional risk stratification for
cardiovascular disease.
• Lipoprotein(a): Independent risk factor for atherosclerosis.

117. Polyunsaturated fatty acids - essential nutritional factors. The role of polyenoic acids as a source of eicosanoids.
Education, biological role, participation of prostaglandins and leukotrienes in the regulation of metabolism and body functions.
Ans. Essential Nature of Polyunsaturated Fatty Acids
Polyunsaturated fatty acids (PUFAs) are fatty acids containing two or more double bonds. The most important nutritionally
essential PUFAs are:
• Linoleic acid (C18:2, ω-6)
• α-Linolenic acid (C18:3, ω-3)
These are termed essential fatty acids because:
• Humans and other mammals cannot synthesize them de novo; they lack the desaturase enzymes to introduce double
bonds beyond the Δ9 position of the fatty acid chain.
• They must be obtained from the diet (mainly from plant oils, nuts, seeds, and fish).
Other important PUFAs, such as arachidonic acid (C20:4, ω-6), can be synthesized from linoleic acid but become essential if
dietary linoleic acid is insufficient15.
Role of Polyenoic Acids as Eicosanoid Precursors
Polyenoic acids (PUFAs with multiple double bonds), especially arachidonic acid, are the main precursors for eicosanoids-a
group of potent, short-lived signaling molecules that include:
• Prostaglandins
• Thromboxanes
• Leukotrienes
Eicosanoid Formation
• Arachidonic acid is released from membrane phospholipids by the action of phospholipase A₂.
• It is then metabolized by:
• Cyclooxygenase (COX) pathway: Produces prostaglandins and thromboxanes.
• Lipoxygenase (LOX) pathway: Produces leukotrienes and other related compounds5.
Biological Role of Eicosanoids
Eicosanoids act as local hormones (autacoids) and participate in the regulation of numerous physiological and pathological
processes:
Prostaglandins
• Regulate inflammation: Mediate fever, pain, and swelling.
• Control vascular tone: Cause vasodilation or vasoconstriction, regulate blood flow.
 • Influence platelet function: Some prostaglandins inhibit platelet aggregation (e.g., prostacyclin), others promote it (e.g.,
thromboxane A₂).
• Modulate gastric secretion: Protect gastric mucosa by inhibiting acid secretion and promoting mucus production.
• Affect smooth muscle: Involved in uterine contractions, bronchial tone, and gastrointestinal motility5.
Leukotrienes
• Mediate allergic and inflammatory responses: Potent bronchoconstrictors (important in asthma), increase vascular
permeability, attract immune cells.
• Participate in immune modulation: Influence leukocyte chemotaxis and activation5.
Participation in Metabolic and Body Function Regulation
• Homeostasis: Eicosanoids help maintain homeostasis by rapidly responding to local stimuli.
• Immunity and Inflammation: Central in both the initiation and resolution of inflammation.
• Cardiovascular Health: Influence blood pressure, clot formation, and vascular health.
• Reproductive System: Regulate ovulation, menstruation, and labor.
• Central Nervous System: Modulate pain perception and fever.

118. Hormonal regulation of the metabolism of carbohydrates, fats and amino acids by insulin. Influence of the nutritional
rhythm on hormonal status.
Ans. Insulin: Central Anabolic Hormone
Insulin is secreted by β-cells of the pancreatic islets in response to increased blood glucose, typically after a meal. It is the key
anabolic hormone regulating the metabolism of carbohydrates, fats, and amino acids.
Effects of Insulin on Metabolism
1. Carbohydrate Metabolism
• Stimulates glucose uptake in muscle and adipose tissue by promoting the translocation of GLUT4 transporters to the
cell membrane.
• Increases glycolysis by activating key enzymes (glucokinase, phosphofructokinase, pyruvate kinase).
• Promotes glycogen synthesis in liver and muscle by activating glycogen synthase and inhibiting glycogen phosphorylase.
• Inhibits gluconeogenesis by repressing the synthesis and activity of enzymes like pyruvate carboxylase, PEP
carboxykinase, fructose-1,6-bisphosphatase, and glucose-6-phosphatase.
• Lowers blood glucose by enhancing cellular uptake and storage, and by suppressing hepatic glucose output12.
2. Lipid Metabolism
• Stimulates fatty acid and triglyceride synthesis in liver and adipose tissue by:
• Activating acetyl-CoA carboxylase and fatty acid synthase.
• Increasing glucose uptake (providing glycerol-3-phosphate for triglyceride synthesis).
• Stimulating lipoprotein lipase (LPL) in adipose tissue, increasing fatty acid uptake from circulating lipoproteins.
• Inhibits lipolysis in adipose tissue by suppressing hormone-sensitive lipase, reducing free fatty acid release.
• Suppresses ketogenesis in the liver by favoring fatty acid storage over oxidation16.
3. Amino Acid and Protein Metabolism
• Stimulates amino acid uptake into cells.
• Promotes protein synthesis by increasing ribosomal activity and gene expression for protein synthesis.
• Inhibits proteolysis (protein breakdown).
 • Promotes synthesis of tissue proteins and enzymes1.
Influence of Nutritional Rhythm on Hormonal Status
Fed (Postprandial) State
• High glucose and amino acid levels after a meal stimulate insulin secretion.
• Insulin predominates, promoting:
• Glucose uptake and storage as glycogen.
• Lipogenesis (fatty acid and triglyceride synthesis).
• Protein synthesis and inhibition of protein breakdown.
• Result: Anabolic processes are favored; blood glucose is lowered and nutrients are stored12.
Fasting (Postabsorptive) State
• Low blood glucose suppresses insulin secretion and stimulates counter-regulatory hormones (glucagon, adrenaline,
cortisol, growth hormone).
• Glucagon predominates, promoting:
• Glycogenolysis and gluconeogenesis in the liver (raising blood glucose).
• Lipolysis in adipose tissue (increasing free fatty acids for energy).
• Ketogenesis in the liver during prolonged fasting.
• Result: Catabolic processes are favored; energy stores are mobilized to maintain blood glucose and supply energy to
vital organs126.
Circadian and Meal-Related Rhythms
• Insulin secretion follows the nutritional rhythm: peaks after meals, lowest during fasting and overnight.
 • Hormonal balance shifts between anabolic (insulin-dominated) and catabolic (counter-regulatory hormone-dominated)
states, coordinating metabolism with nutrient availability4.

119. Biological oxidation. The main stages of the unification of the energy material. Oxidative processes are the main sources
of NADH. Intramitochondrial and extramitochondrial sources of NADH.
Ans. 1. Biological Oxidation: Definition and Overview
Biological oxidation refers to the enzyme-catalyzed processes by which cells extract energy from nutrients through the transfer
of electrons (or hydrogen atoms) to molecular oxygen or other acceptors. This process is fundamental for ATP synthesis and
cellular energy metabolism14.
2. Main Stages of the Unification of Energy Material
The catabolic breakdown of carbohydrates, fats, and proteins proceeds through a series of unified stages, ultimately leading to
the production of ATP:
Stage 1: Digestion (Primary Metabolism)
• Location: Gastrointestinal tract.
• Process: Large macromolecules (proteins, polysaccharides, lipids) are broken down into their monomeric units (amino
acids, monosaccharides, fatty acids, glycerol).
• Energy: Minimal ATP production; most energy is lost as heat1.
Stage 2: Formation of Common Intermediates (Secondary Metabolism)
• Location: Cytosol and mitochondrial matrix.
• Process: Monomers are converted to a limited set of key metabolites, mainly acetyl-CoA and a few carboxylic acids (e.g.,
pyruvate, oxaloacetate, malate).
 • Energy: Some ATP and reducing equivalents (NADH, FADH₂) are generated1.
Stage 3: Complete Oxidation in the Citric Acid (Krebs) Cycle (Tertiary Metabolism)
• Location: Mitochondrial matrix.
• Process: Acetyl-CoA enters the Krebs cycle, where it is fully oxidized to CO₂. The cycle’s main function is to concentrate
metabolic energy as reduced coenzymes (NADH, FADH₂)14.
• Outcome: The specificity of the original substrate is lost; all fuels are funneled into a common pathway.
Stage 4: Terminal Oxidation (Electron Transport Chain & Oxidative Phosphorylation)
• Location: Inner mitochondrial membrane.
• Process: NADH and FADH₂ donate electrons to the electron transport chain (ETC), which ultimately reduces O₂ to H₂O.
The energy released is used to generate a proton gradient, driving ATP synthesis by ATP synthase14.
• Outcome: ATP is produced; water and CO₂ are the final waste products.
3. Oxidative Processes as Main Sources of NADH
NADH is the principal carrier of electrons (reducing equivalents) from catabolic pathways to the mitochondrial electron
transport chain.
Major NADH-Producing Processes:
• Glycolysis (Cytosol):
• Oxidation of glyceraldehyde-3-phosphate to 1,3-bisphosphoglycerate (enzyme: glyceraldehyde-3-phosphate
dehydrogenase) produces NADH24.
• Pyruvate Dehydrogenase Complex (Mitochondria):
• Oxidative decarboxylation of pyruvate to acetyl-CoA produces NADH24.
 • Citric Acid Cycle (Mitochondria):
• Several dehydrogenase reactions produce NADH:
• Isocitrate dehydrogenase
• α-Ketoglutarate dehydrogenase
• Malate dehydrogenase24.
• β-Oxidation of Fatty Acids (Mitochondria):
• Each cycle produces one NADH per turn34.
• Amino Acid Catabolism:
• Certain dehydrogenase steps in amino acid breakdown (e.g., glutamate dehydrogenase) produce NADH4.
4. Intramitochondrial and Extramitochondrial Sources of NADH
A. Intramitochondrial Sources (Within Mitochondria):
• Pyruvate dehydrogenase complex
• Citric acid cycle dehydrogenases
• β-oxidation of fatty acids
• Amino acid oxidation
• Malate dehydrogenase (mitochondrial form)
These NADH molecules directly feed electrons into the mitochondrial electron transport chain4.
B. Extramitochondrial (Cytosolic) Sources:
• Glycolysis:
 • Glyceraldehyde-3-phosphate dehydrogenase reaction produces NADH in the cytosol24.
• Other cytosolic dehydrogenases:
• e.g., lactate dehydrogenase, alcohol dehydrogenase.
Note: NADH cannot cross the inner mitochondrial membrane directly. Instead, its reducing equivalents are shuttled into the
mitochondria via:
• Malate-aspartate shuttle: Active in liver, kidney, heart.
• Glycerol-3-phosphate shuttle: Active in skeletal muscle and brain14.
These shuttles transfer electrons from cytosolic NADH to mitochondrial NAD⁺ or FAD, enabling their use in oxidative
phosphorylation.

120. Shuttle enzyme-substrate systems for the transfer of hydrogen from the cytoplasm to mitochondria. The value of the
process.
Ans. Why Are Shuttle Systems Needed?
During glycolysis in the cytoplasm, NADH is produced (notably by glyceraldehyde-3-phosphate dehydrogenase). However,
the inner mitochondrial membrane is impermeable to NADH. For the electrons (reducing equivalents) from cytosolic NADH to
be used in the mitochondrial electron transport chain (ETC) for ATP production, they must be transferred into the
mitochondria by shuttle systems1.
Main Shuttle Systems
1. Malate-Aspartate Shuttle
• Location: Predominant in liver, kidney, and heart.
• Mechanism:
 • In the cytosol, NADH reduces oxaloacetate to malate (via cytosolic malate dehydrogenase).
• Malate is transported into the mitochondrial matrix via the malate-α-ketoglutarate transporter.
• In the matrix, malate is oxidized back to oxaloacetate (via mitochondrial malate dehydrogenase), regenerating
NADH inside the mitochondria.
• Oxaloacetate is transaminated to aspartate, which is transported back to the cytosol, where it is converted back to
oxaloacetate, completing the cycle.
• ATP Yield: Each cytosolic NADH yields about 3 ATP (or 2.5 ATP, depending on the system)1.
2. Glycerol 3-Phosphate Shuttle
• Location: Predominant in skeletal muscle and brain.
• Mechanism:
• In the cytosol, NADH reduces dihydroxyacetone phosphate (DHAP) to glycerol 3-phosphate (via cytosolic glycerol
3-phosphate dehydrogenase).
• Glycerol 3-phosphate is oxidized back to DHAP by a mitochondrial membrane-bound glycerol 3-phosphate
dehydrogenase, transferring electrons to FAD (forming FADH₂), which then passes electrons to ubiquinone (CoQ)
in the ETC.
• ATP Yield: Each cytosolic NADH yields about 2 ATP (or 1.5 ATP, as FADH₂ enters the ETC at Complex II, bypassing Complex
I)1.
Other Shuttle Systems (Less Prominent)
• α-Glycerophosphate Shuttle
• Lactate Shuttle
• Glutamate Shuttle
 • These play minor or tissue-specific roles and are variations or related to the main two shuttles above1.
Value and Biological Significance of Shuttle Systems
• Efficient Energy Production: They enable the energy stored in cytosolic NADH (from glycolysis) to be used for ATP
synthesis in mitochondria.
• Tissue Specificity: Different tissues use different shuttles based on their metabolic needs. For example, the malate-
aspartate shuttle is more energy-efficient and is used in tissues with high aerobic capacity (heart, liver), while the
glycerol 3-phosphate shuttle is used in tissues requiring rapid ATP generation (muscle, brain)1.
• Maintaining Redox Balance: Shuttles help regenerate NAD⁺ in the cytosol, which is essential for continuous glycolytic
flux.
• Metabolic Flexibility: Allow cells to adapt to varying energy demands and oxygen availability.

121. Catabolism and anabolism and their relationship in metabolism. Endergonic and exergonic reactions in metabolism. ATP
and other macroergs. ATP is a universal chemical form of energy storage in the cell. ADP-ATP cycle. Oxidative and substrate
phosphorylation. Use of ATP.
Ans. Metabolism is the sum of all chemical reactions in the cell, divided into two complementary processes:
• Catabolism: The degradative phase. Large, complex molecules (carbohydrates, fats, proteins) are broken down into
smaller, simpler molecules (e.g., CO₂, H₂O, NH₃, lactic acid). Catabolic pathways release energy, some of which is
captured in the form of ATP, NADH, NADPH, and FADH₂, while the rest is lost as heat16.
• Anabolism: The biosynthetic phase. Small, simple precursors are built up into larger, more complex molecules (lipids,
polysaccharides, proteins, nucleic acids). Anabolic pathways require energy, which they obtain from ATP and reducing
equivalents (NADPH, NADH, FADH₂) produced during catabolism16.
Relationship:
Catabolic pathways supply the energy and building blocks required for anabolic processes. The energy released from
catabolism is conserved in high-energy molecules (mainly ATP) and reducing equivalents, which are then used to drive
endergonic (energy-consuming) anabolic reactions1.
Endergonic and Exergonic Reactions in Metabolism
• Exergonic reactions: Release free energy (ΔG < 0). These are typically catabolic reactions, such as the oxidation of
glucose, fatty acids, or amino acids. The energy released is harnessed for cellular work1.
• Endergonic reactions: Require an input of free energy (ΔG > 0). These are typically anabolic reactions, such as synthesis
of proteins, nucleic acids, and complex carbohydrates1.
• Coupling: In metabolism, exergonic and endergonic reactions are often coupled, so the energy released from exergonic
reactions is used to drive endergonic processes, frequently via ATP1.
ATP and Other Macroergs
• ATP (Adenosine Triphosphate): The primary "energy currency" of the cell. It consists of adenine, ribose, and three
phosphate groups. The bonds between the phosphate groups (especially the terminal two) are "macroergic" (high-
energy) bonds1.
• Other macroergs: Include GTP, CTP, UTP, creatine phosphate, and acetyl-CoA, which also store and transfer energy in
specific metabolic contexts1.
ATP is not a long-term energy store (that role belongs to glycogen and fat), but rather a universal, rapidly mobilizable energy
carrier that shuttles energy to where it is needed in the cell1.
ATP as a Universal Chemical Form of Energy Storage
• ATP is constantly synthesized and consumed in all living cells. It acts as a universal intermediary, capturing energy from
exergonic reactions and providing it for endergonic processes such as biosynthesis, active transport, muscle contraction,
and cell signaling1.
 • ATP’s high-energy phosphate bonds are hydrolyzed to release energy, converting ATP to ADP (adenosine diphosphate)
or AMP (adenosine monophosphate)1.
The ADP-ATP Cycle
• Cycle: ATP is hydrolyzed to ADP (and Pi), releasing energy for cellular work. ADP is then re-phosphorylated to ATP using
energy from catabolic reactions (e.g., oxidative phosphorylation, substrate-level phosphorylation)1.
• Continuous turnover: The average human recycles their body weight in ATP daily, due to this rapid cycling1.
Oxidative and Substrate-Level Phosphorylation
• Substrate-level phosphorylation: ATP is formed directly by the transfer of a phosphate group from a high-energy
substrate to ADP. This occurs in glycolysis (e.g., 1,3-bisphosphoglycerate to 3-phosphoglycerate) and in the citric acid
cycle (succinyl-CoA to succinate)2.
• Oxidative phosphorylation: ATP is synthesized from ADP and Pi, driven by the transfer of electrons from NADH and
FADH₂ to O₂ via the electron transport chain, creating a proton gradient across the inner mitochondrial membrane. ATP
synthase uses this gradient to phosphorylate ADP12.
Use of ATP
ATP is used for:
• Biosynthetic reactions: Protein, nucleic acid, lipid, and polysaccharide synthesis.
• Active transport: Pumping ions and molecules across membranes (e.g., Na⁺/K⁺-ATPase).
• Mechanical work: Muscle contraction, ciliary and flagellar movement.
• Cell signaling: Phosphorylation of proteins and secondary messenger systems.
• Heat production: In brown adipose tissue and during uncoupled respiration12.
122. Structural organization of the electron and proton transport chain. ATP synthetase, ATP synthesis. The relationship
between oxidation and phosphorylation.
Ans. 1. Structural Organization of the Electron and Proton Transport Chain (ETC)
The electron transport chain (ETC) is located in the inner mitochondrial membrane and is composed of a series of protein
complexes and mobile carriers that transfer electrons from reduced coenzymes (NADH, FADH₂) to molecular oxygen, coupled
with the translocation of protons (H⁺) across the membrane.
Main Components of the ETC:
• Complex I (NADH:ubiquinone oxidoreductase, NADH dehydrogenase):
• Accepts electrons from NADH, transfers them to ubiquinone (CoQ), and pumps 4 protons from the matrix to the
intermembrane space per electron pair.
• Contains FMN and iron-sulfur (Fe-S) clusters3.
• Complex II (Succinate:ubiquinone oxidoreductase, succinate dehydrogenase):
• Accepts electrons from FADH₂ (produced in the TCA cycle), transfers them to CoQ. Does not pump protons3.
• Ubiquinone (Coenzyme Q, Q):
• A lipid-soluble mobile carrier that shuttles electrons from Complexes I and II to Complex III within the
membrane3.
• Complex III (Ubiquinol:cytochrome c oxidoreductase, cytochrome bc₁ complex):
• Transfers electrons from reduced CoQ (QH₂) to cytochrome c, pumps 4 protons per electron pair3.
• Cytochrome c:
• A small, water-soluble protein that carries electrons from Complex III to Complex IV3.
• Complex IV (Cytochrome c oxidase):
 • Transfers electrons from cytochrome c to molecular oxygen (O₂), reducing it to water, and pumps 2 protons per
electron pair3.
Proton Gradient:
• As electrons flow through Complexes I, III, and IV, protons are pumped from the mitochondrial matrix to the
intermembrane space, creating an electrochemical (proton-motive) gradient across the inner membrane3.
2. ATP Synthase (Complex V) and ATP Synthesis
ATP synthase is a large enzyme complex embedded in the inner mitochondrial membrane, composed of two main parts:
• F₀ subunit: Forms a proton channel spanning the membrane, allowing protons to flow back into the matrix.
• F₁ subunit: Projects into the matrix and contains the catalytic sites for ATP synthesis3.
Mechanism of ATP Synthesis:
• The proton-motive force (electrochemical gradient of H⁺) generated by the ETC drives protons through the F₀ channel of
ATP synthase.
• This proton flow causes rotation of the F₀ subunit and conformational changes in the F₁ subunit, catalyzing the
phosphorylation of ADP to ATP (rotational catalysis/Boyer’s binding change mechanism)3.
• For every full rotation, three ATP molecules are synthesized3.
3. The Relationship Between Oxidation and Phosphorylation (Oxidative Phosphorylation)
• Oxidation: Refers to the transfer of electrons from NADH and FADH₂ through the ETC to O₂, which is reduced to H₂O3.
• Phosphorylation: Refers to the synthesis of ATP from ADP and inorganic phosphate (Pi), catalyzed by ATP synthase3.
• Coupling: The two processes are tightly coupled. The energy released by electron transfer is used to pump protons and
establish the proton gradient, which in turn powers ATP synthesis3.
 • Chemiosmotic Theory: Proposed by Peter Mitchell, this theory explains that the energy from electron transfer is
conserved by pumping protons across the membrane, and ATP synthesis is driven by the return flow of protons through
ATP synthase3.
Regulation:
• The rate of oxidative phosphorylation is controlled by the availability of ADP (acceptor control) and the proton gradient.
If ADP is limited, electron flow and oxygen consumption decrease3.
Uncoupling:
• If the proton gradient is dissipated by uncouplers (e.g., dinitrophenol), electron transport and oxygen consumption
continue, but ATP synthesis is diminished or stopped, and energy is released as heat3.

123. Chemiosmotic theory of oxidative phosphorylation by Mitchell-Skulachev.

Ans. Essence of the Chemiosmotic Theory
The chemiosmotic theory, formulated by Peter Mitchell (1961) and further developed by Vladimir Skulachev, explains how the
energy from electron transfer in the mitochondrial respiratory chain is used to synthesize ATP during oxidative
phosphorylation.
Key Concepts:
• Electron Transport and Proton Pumping:
As electrons are transferred through the complexes of the electron transport chain (ETC) in the inner mitochondrial
membrane (Complexes I, III, and IV), the energy released is used to pump protons (H⁺) from the mitochondrial matrix
into the intermembrane space1.
• Generation of Proton-Motive Force:
This proton pumping creates an electrochemical gradient across the inner membrane, consisting of:
 • A chemical (pH) gradient (matrix becomes more alkaline)
• An electrical gradient (matrix becomes more negative relative to the intermembrane space)
Together, these form the proton-motive force (PMF)1.
• ATP Synthesis Driven by Proton Flow:
The inner mitochondrial membrane is impermeable to protons except via the enzyme ATP synthase (Complex V).
Protons flow back into the matrix through ATP synthase, and the energy released by this flow is used to convert ADP and
Pi into ATP1.
• Coupling of Oxidation and Phosphorylation:
The process of electron transfer (oxidation) is thus tightly coupled to ATP synthesis (phosphorylation) by the proton
gradient1.
Mitchell-Skulachev Model: Mechanistic Details
• Respiratory Chain Complexes as Proton Pumps:
Complexes I, III, and IV act as proton pumps, translocating protons from the matrix to the intermembrane space as
electrons pass through the ETC1.
• Proton-Motive Force (Δp):
The energy stored in the proton gradient (Δp) is the sum of the membrane potential (Δψ) and the pH gradient (ΔpH).
This energy is harnessed by ATP synthase1.
• ATP Synthase Function:
ATP synthase consists of two main components:
• F₀ subunit: Forms a proton channel across the membrane.
• F₁ subunit: Catalyzes ATP synthesis in the matrix.
As protons flow through F₀, conformational changes in F₁ drive the phosphorylation of ADP to ATP (rotational
catalysis)1.
 • Regulation:
The rate of oxidative phosphorylation is regulated by the availability of ADP (acceptor control) and the proton gradient.
If the gradient is dissipated (e.g., by uncouplers), electron transport continues but ATP synthesis ceases, and energy is
released as heat1.
Experimental and Clinical Significance
• Experimental Support:
• Artificially imposed pH gradients can drive ATP synthesis even in the absence of electron flow, confirming the
central role of the proton gradient.
• Uncouplers (e.g., dinitrophenol) collapse the proton gradient, abolishing ATP synthesis without stopping electron
flow1.
• Clinical Relevance:
• Defects in the ETC or ATP synthase, or the presence of uncouplers, lead to impaired ATP production and various
mitochondrial diseases.

124. Terminal phase of biological oxidation. Mitochondrial electron transport chain. Mechanism of oxidative phosphorylation.
Ans. Terminal Phase of Biological Oxidation
The terminal phase of biological oxidation is the final stage of cellular respiration where the energy stored in reduced
coenzymes (NADH, FADH₂), produced during glycolysis, the citric acid (Krebs) cycle, and β-oxidation, is used to synthesize ATP.
This occurs in mitochondria and is known as oxidative phosphorylation14.
Mitochondrial Electron Transport Chain (ETC)
The ETC is a series of protein complexes and mobile carriers embedded in the inner mitochondrial membrane. Its main
function is to transfer electrons from NADH and FADH₂ to molecular oxygen (O₂), reducing it to water (H₂O), while
simultaneously pumping protons (H⁺) from the mitochondrial matrix into the intermembrane space, creating a proton
gradient14.
Key Components and Sequence:
1. Complex I (NADH-CoQ reductase/NADH dehydrogenase):
• Accepts electrons from NADH, transfers them to ubiquinone (CoQ), and pumps 4 protons into the intermembrane
space.
2. Complex II (Succinate-CoQ reductase/succinate dehydrogenase):
• Accepts electrons from FADH₂ (from succinate oxidation), transfers them to CoQ. Does not pump protons.
3. Ubiquinone (Coenzyme Q):
• Lipid-soluble carrier, shuttles electrons from Complexes I and II to Complex III.
4. Complex III (CoQ-cytochrome c reductase/cytochrome bc₁ complex):
• Transfers electrons from CoQH₂ to cytochrome c, pumps 4 protons.
5. Cytochrome c:
• Small, soluble protein, transfers electrons from Complex III to Complex IV.
6. Complex IV (Cytochrome c oxidase):
• Transfers electrons from cytochrome c to O₂, reducing it to H₂O, and pumps 2 protons.
Summary of Proton Movement:
• Complexes I, III, and IV act as proton pumps, generating an electrochemical (proton-motive) gradient across the inner
mitochondrial membrane1.
Mechanism of Oxidative Phosphorylation
The chemiosmotic theory (Mitchell-Skulachev) explains how the energy from electron transfer is used to synthesize ATP1:
1. Proton Gradient Formation:
• Electron flow through the ETC drives protons from the matrix to the intermembrane space, creating a proton-
motive force (difference in pH and electric potential).
2. ATP Synthesis by ATP Synthase (Complex V):
• ATP synthase is a large enzyme complex with two main parts:
• F₀ subunit: Forms a proton channel across the membrane.
• F₁ subunit: Catalyzes ATP synthesis in the matrix.
• Protons flow back into the matrix through F₀, causing conformational changes in F₁ that catalyze the
phosphorylation of ADP to ATP (rotational catalysis)1.
3. Coupling of Oxidation and Phosphorylation:
• The processes of electron transport (oxidation) and ATP synthesis (phosphorylation) are tightly coupled; ATP
synthesis depends on the proton gradient generated by the ETC.
• The rate of oxidative phosphorylation is regulated by the availability of ADP (acceptor control)1.
4. Stoichiometry:
• Each NADH yields about 2.5–3 ATP; each FADH₂ yields about 1.5–2 ATP, reflecting their entry points into the chain
and the number of protons pumped1.
Clinical and Functional Notes
• Uncouplers (e.g., dinitrophenol) dissipate the proton gradient, allowing electron transport and oxygen consumption to
continue without ATP synthesis, resulting in heat production.
 • ETC inhibitors (e.g., cyanide, CO) block electron flow, halting both ATP synthesis and oxygen consumption, which is
fatal1.
• Inherited mitochondrial disorders affecting ETC complexes can lead to severe energy deficiency, especially in tissues
with high energy demands (muscle, brain)1.

125. Respiratory control. Dissociation of respiration and phosphorylation. Hypoenergetic states.

Ans. Respiratory Control
Respiratory control refers to the regulation of the rate of mitochondrial respiration (oxygen consumption and electron
transport) by the availability of ADP and inorganic phosphate (Pi). This is a key mechanism ensuring that ATP production
matches the cell's energy demand.
• Mechanism:
• When ADP levels are high (i.e., more ATP is being used), the electron transport chain (ETC) and oxidative
phosphorylation accelerate to replenish ATP.
• When ADP is low (ATP demand is low), ETC activity slows because the proton gradient is not being dissipated by
ATP synthase, leading to a buildup of proton-motive force and inhibition of further electron flow.
• Biological significance:
• This coupling ensures efficient energy production, prevents wasteful oxidation of substrates, and protects cells
from excessive reactive oxygen species (ROS) generation25.
Dissociation of Respiration and Phosphorylation (Uncoupling)
Dissociation (uncoupling) of respiration and phosphorylation occurs when electron transport and oxygen consumption
proceed without the synthesis of ATP. This can happen due to:
• Chemical uncouplers:
 • Compounds like 2,4-dinitrophenol (DNP), FCCP, and CCCP increase the permeability of the inner mitochondrial
membrane to protons, dissipating the proton gradient without driving ATP synthase2.
• Physiological uncoupling:
• Uncoupling proteins (e.g., UCP1 in brown adipose tissue) allow protons to re-enter the mitochondrial matrix
without ATP synthesis, generating heat instead (non-shivering thermogenesis).
• Consequence:
• Oxygen consumption and substrate oxidation increase, but ATP is not produced; energy is released as heat.
• This can protect against excessive ROS but, if uncontrolled, leads to energy deficiency despite high oxygen
consumption2.
Hypoenergetic States
Hypoenergetic states are conditions in which ATP production is insufficient to meet cellular energy demands. They can result
from:
• Defects in respiratory control or coupling:
• Uncoupling (chemical or genetic) leads to inadequate ATP despite active respiration.
• Inhibition of the ETC or ATP synthase:
• Poisons like cyanide, CO, or oligomycin block electron flow or ATP synthesis, halting energy production.
• Mitochondrial dysfunction:
• Genetic or acquired defects in mitochondrial enzymes or membrane integrity reduce ATP output.
• Clinical consequences:
• Manifest as muscle weakness, neurological deficits, lactic acidosis (due to increased anaerobic glycolysis), organ
dysfunction, and in severe cases, death2.
126. Microsomal oxidation. The main enzymes of the microsomal electron transport chain. Biological role.
Ans. What is Microsomal Oxidation?
Microsomal oxidation refers to a set of oxidative reactions that occur in the smooth endoplasmic reticulum (ER) of cells,
especially in the liver. These reactions are catalyzed by a group of enzymes collectively known as the microsomal electron
transport chain. The system is crucial for the metabolism of a wide variety of endogenous compounds (such as steroids, fatty
acids, and bilirubin) and exogenous substances (drugs, toxins, and carcinogens).
Main Enzymes of the Microsomal Electron Transport Chain
The key enzyme system is the cytochrome P450 monooxygenase system, which consists of:
1. Cytochrome P450 enzymes (CYPs)
• A large family of heme-containing monooxygenases.
• Catalyze the insertion of one oxygen atom into a substrate (RH) and the other into water:
RH+O2+NADPH+H+→ROH+H2O+NADP+RH+O2+NADPH+H+→ROH+H2O+NADP+
• There are over 150 forms of CYPs, grouped into families with diverse substrate specificities1.
2. NADPH-cytochrome P450 reductase
• A flavoprotein containing FAD and FMN.
• Transfers electrons from NADPH to cytochrome P450, enabling the monooxygenase reaction1.
3. Cytochrome b5 and NADH-cytochrome b5 reductase
• Cytochrome b5 can participate in electron transfer, especially in fatty acid desaturation.
• NADH-cytochrome b5 reductase transfers electrons from NADH to cytochrome b51.
Mechanism of Microsomal Oxidation
• Electron flow:
NADPH (or NADH) → NADPH-cytochrome P450 reductase (FAD, FMN) → cytochrome P450 (heme) → substrate
oxidation1.
• Monooxygenase reaction:
Incorporates one atom of molecular oxygen into the substrate (hydroxylation), the other is reduced to water.
Biological Role
1. Detoxification (Biotransformation) of Xenobiotics
• Phase I reactions:
• Hydroxylation, oxidation, reduction, and dealkylation of drugs, toxins, and carcinogens to make them more polar
and easier to excrete.
• Phase II reactions:
• The products of phase I are further conjugated with glucuronic acid, sulfate, glutathione, or other groups to
enhance solubility and elimination1.
2. Metabolism of Endogenous Compounds
• Steroid hormone synthesis and metabolism
• Bile acid synthesis
• Fatty acid and eicosanoid metabolism
• Vitamin D activation and inactivation
3. Regulation of Drug Activity
• Influences drug half-life, activity, and toxicity.
 • Responsible for drug-drug interactions via induction or inhibition of specific CYP isoforms.
4. Fatty Acid Desaturation
• Cytochrome b5 and its reductase are important for the introduction of double bonds into fatty acids1.
127. Prooxidant processes. Formation of reactive oxygen species, representatives, mechanism of the damaging action of
biomolecules and structures.
Ans. Prooxidant Processes and Formation of Reactive Oxygen Species (ROS)
Prooxidant processes are metabolic or chemical reactions that lead to the formation of highly reactive oxygen-containing
molecules, collectively known as reactive oxygen species (ROS). These are primarily generated during normal cellular
metabolism, especially in the mitochondria during oxidative phosphorylation, as well as in microsomal oxidation and during
immune responses.
Main Sources of ROS:
• Mitochondrial electron transport chain (Complexes I and III)
• Microsomal cytochrome P450 system
• Enzymatic reactions (e.g., xanthine oxidase, NADPH oxidase in phagocytes)
• Ionizing radiation, drugs, toxins, and environmental pollutants
Key Representatives of Reactive Oxygen Species
1. Superoxide anion (O₂- ⁻):
• Formed by the univalent (one-electron) reduction of molecular oxygen.
• Highly reactive and can initiate further ROS formation.
2. Hydrogen peroxide (H₂O₂):
 • Produced by dismutation of superoxide (via superoxide dismutase) or directly by oxidases.
• Less reactive than superoxide but can cross membranes and generate more dangerous radicals.
3. Hydroxyl radical (- OH):
• Generated from H₂O₂ via the Fenton reaction (in the presence of Fe²⁺ or Cu⁺).
• Extremely reactive, capable of damaging almost all types of biomolecules.
4. Singlet oxygen (¹O₂):
• An excited form of oxygen with high reactivity, produced during photochemical reactions.
5. Peroxyl and alkoxyl radicals (ROO- , RO- ):
• Formed during lipid peroxidation chain reactions.
6. Hydroperoxyl radical (HO₂- ):
• A protonated form of superoxide, also reactive.
7. Nitric oxide (NO- ) and peroxynitrite (ONOO⁻):
• Reactive nitrogen species (RNS) that interact with ROS, amplifying damage.
Mechanism of Damaging Action on Biomolecules and Structures
ROS are highly reactive and can cause oxidative damage to all classes of biomolecules, leading to cellular dysfunction and
death.
1. Lipid Peroxidation
• Target: Polyunsaturated fatty acids in membrane phospholipids.
• Process: Initiation by hydroxyl radicals leads to formation of lipid radicals, propagation of chain reactions, and formation
of lipid peroxides and aldehydes (e.g., malondialdehyde).
 • Consequence: Loss of membrane integrity, increased permeability, impaired membrane-bound enzymes and receptors.
2. Protein Oxidation
• Target: Amino acid side chains, peptide backbone.
• Process: Oxidation of thiol groups, aromatic residues, and formation of protein carbonyls.
• Consequence: Enzyme inactivation, altered structural proteins, increased proteolysis, aggregation.
3. DNA Damage
• Target: DNA bases and sugar-phosphate backbone.
• Process: Hydroxyl radicals cause strand breaks, base modifications (e.g., 8-oxoguanine), cross-linking.
• Consequence: Mutations, impaired replication and transcription, carcinogenesis, cell death.
4. Carbohydrate and Other Molecule Damage
• Target: Glycosaminoglycans, monosaccharides.
• Process: Fragmentation, cross-linking, loss of function.
5. Cellular and Tissue Effects
• Membrane damage: Increased permeability, cell lysis.
• Mitochondrial dysfunction: Impaired ATP synthesis, apoptosis.
• Activation of cell death pathways: Necrosis, apoptosis, or autophagy.
Clinical and Biological Significance
• Aging: Accumulation of oxidative damage is implicated in aging.
• Disease: ROS are involved in the pathogenesis of atherosclerosis, neurodegenerative diseases (Alzheimer’s, Parkinson’s),
diabetes, cancer, chronic inflammation, ischemia-reperfusion injury.
 • Immune defense: Controlled ROS production in phagocytes is essential for killing pathogens (respiratory burst).
Antioxidant Defense Systems
Cells possess enzymatic and non-enzymatic antioxidant systems to neutralize ROS:
• Enzymatic: Superoxide dismutase (SOD), catalase, glutathione peroxidase.
• Non-enzymatic: Vitamin E, vitamin C, glutathione, carotenoids, flavonoids, uric acid.

128. Antioxidant defense system. Antioxidant enzymes. Low molecular weight antioxidants.
Ans. The antioxidant defense system protects cells and tissues from damage caused by reactive oxygen species (ROS) and
other free radicals. This system includes both specialized enzymes and a range of low molecular weight (non-enzymatic)
antioxidants.
Antioxidant Enzymes
These enzymes catalytically remove or neutralize reactive oxygen species, preventing oxidative damage:
1. Superoxide Dismutase (SOD)
• Function: Converts superoxide anion (O₂- ⁻) into hydrogen peroxide (H₂O₂) and oxygen.
• Reaction:
2O2−−+2H+→H2O2+O22O2−−+2H+→H2O2+O2
• Location: Present in cytosol (Cu/Zn-SOD), mitochondria (Mn-SOD), and extracellular spaces.
2. Catalase
• Function: Decomposes hydrogen peroxide into water and oxygen, preventing the formation of the highly toxic hydroxyl
radical.
 • Reaction:
2H2O2→2H2O+O22H2O2→2H2O+O2
• Location: Mainly in peroxisomes.
3. Glutathione Peroxidase (GPx)
• Function: Reduces hydrogen peroxide and organic hydroperoxides to water or corresponding alcohols, using reduced
glutathione (GSH) as an electron donor.
• Reaction:
2GSH+H2O2→GSSG+2H2O2GSH+H2O2→GSSG+2H2O
• Location: Cytosol and mitochondria; contains selenium as a cofactor.
4. Glutathione Reductase
• Function: Regenerates reduced glutathione (GSH) from its oxidized form (GSSG), using NADPH.
• Reaction:
GSSG+NADPH+H+→2GSH+NADP+GSSG+NADPH+H+→2GSH+NADP+
5. Other Enzymes
• Peroxidases: Catalyze the reduction of peroxides.
• Ceruloplasmin: Acts as an antioxidant by binding copper and scavenging free radicals.
Low Molecular Weight (Non-Enzymatic) Antioxidants
These molecules directly scavenge ROS or regenerate other antioxidants:
1. Vitamin E (α-Tocopherol)
• Lipid-soluble antioxidant found in cell membranes.
 • Protects polyunsaturated fatty acids from lipid peroxidation by scavenging lipid radicals.
2. Vitamin C (Ascorbic Acid)
• Water-soluble antioxidant in plasma and cytosol.
• Regenerates vitamin E from its oxidized form and neutralizes a variety of ROS.
3. Glutathione (GSH)
• Tripeptide (γ-glutamylcysteinylglycine) present in high concentrations in cells.
• Directly scavenges ROS and acts as a substrate for glutathione peroxidase.
4. Carotenoids (e.g., β-Carotene, Lycopene)
• Quench singlet oxygen and scavenge peroxyl radicals.
• Found in vegetables and fruits (e.g., carrots, tomatoes).
5. Flavonoids and Polyphenols
• Plant-derived antioxidants that neutralize free radicals and chelate metal ions.
6. Uric Acid
• Endogenous antioxidant in plasma, scavenges singlet oxygen and peroxyl radicals.
7. Bilirubin
• Product of heme degradation with antioxidant properties.
8. Trace Elements
• Selenium: Essential for glutathione peroxidase activity.
• Zinc, Copper, Manganese: Cofactors for SOD isoforms.
Classification by Mode of Action
• Preventive antioxidants: Reduce the rate of chain initiation (e.g., catalase, peroxidases, metal ion chelators).
• Chain-breaking antioxidants: Interrupt propagation of free radical chain reactions (e.g., vitamin E, SOD, uric acid).
Clinical Importance
• Antioxidant defenses are critical for protecting against oxidative damage implicated in aging, atherosclerosis, cancer,
diabetes, neurodegenerative diseases, and inflammatory conditions.
• Deficiency or dysfunction in antioxidant systems increases susceptibility to oxidative stress and related diseases12.

129. Minerals as micronutrients. Sources and need. General functions of minerals.

Ans. What Are Minerals?
Minerals are inorganic elements essential for the body's biochemical and physiological processes. As micronutrients, they are
required in small amounts compared to macronutrients (proteins, fats, carbohydrates), but are vital for health, growth, and
metabolism. Minerals are classified as macrominerals (needed in larger amounts, e.g., calcium, magnesium, sodium,
potassium, phosphorus, chloride, sulfur) and trace elements or microminerals (needed in minute amounts, e.g., iron, zinc,
copper, iodine, selenium, manganese, fluoride, chromium, molybdenum, cobalt)1.
Sources of Minerals
• Dietary Sources:
• Animal products: Meat, fish, eggs, milk, cheese (rich in calcium, phosphorus, iron, zinc, iodine, selenium).
• Plant products: Vegetables, fruits, whole grains, legumes, nuts, seeds (sources of magnesium, potassium, iron,
manganese, copper).
• Water: Some minerals (e.g., fluoride, calcium, magnesium) are also present in drinking water.
 • Bioavailability:
• The fraction of ingested mineral that is absorbed and utilized depends on the mineral's chemical form, presence
of enhancers (e.g., vitamin C for iron) or inhibitors (e.g., phytates, oxalates), and physiological state (e.g., growth,
pregnancy, illness)1.
Need for Minerals
• Daily Requirement:
• The amount needed varies by mineral, age, sex, physiological state, and health status.
• Requirements are defined as Recommended Dietary Allowances (RDAs) or Dietary Reference Intakes (DRIs),
which are set to meet the needs of nearly all healthy individuals1.
• For example:
• Calcium: 1000–1300 mg/day (adults)
• Iron: 8–18 mg/day (adults, higher for women)
• Zinc: 8–11 mg/day
• Iodine: 150 µg/day
• Selenium: 55 µg/day
General Functions of Minerals
Minerals perform diverse and essential roles in the body, including:
• Structural Functions:
• Calcium and phosphorus are main components of bones and teeth.
• Magnesium is part of bone structure and stabilizes ATP.
• Regulation of Fluid and Electrolyte Balance:
• Sodium, potassium, and chloride maintain osmotic balance, acid-base status, and are crucial for nerve impulse
transmission and muscle contraction.
• Cofactors for Enzymes:
• Many minerals act as essential cofactors for enzymes (e.g., zinc for DNA/RNA polymerases, magnesium for
kinases, selenium for glutathione peroxidase, copper for cytochrome oxidase, iron for catalase and peroxidase)6.
• Oxygen Transport and Storage:
• Iron is a component of hemoglobin and myoglobin, essential for oxygen transport and storage.
• Copper is required for iron metabolism and hemoglobin synthesis.
• Hormone Synthesis and Action:
• Iodine is required for thyroid hormone synthesis.
• Zinc is necessary for insulin storage and release.
• Antioxidant Defense:
• Selenium is a component of antioxidant enzymes (e.g., glutathione peroxidase).
• Zinc and copper are cofactors for superoxide dismutase.
• Transmission of Nerve Impulses and Muscle Function:
• Calcium, potassium, sodium, and magnesium are essential for nerve conduction and muscle contraction,
including heart rhythm.
• Immune Function:
• Zinc, selenium, iron, and copper support immune cell development and function.
130. The specific role of sodium, potassium, chlorine ions in the vital activity of the organism. Potassium, sodium, blood
chlorides. Hypo- and hypernatremia, hypo- and hyperkalemia.
Ans. Physiological Roles of Na⁺, K⁺, and Cl⁻ Ions
Sodium (Na⁺)
• Principal extracellular cation.
• Maintains extracellular fluid (ECF) volume and osmotic pressure: Sodium is the main determinant of ECF osmolarity
and thus regulates blood volume and blood pressure.
• Nerve impulse transmission: Na⁺ influx is essential for the depolarization phase of action potentials in nerves and
muscles.
• Acid-base balance: Participates in bicarbonate reabsorption in the kidneys.
• Nutrient absorption: Co-transport with glucose and amino acids in the intestine and renal tubules.
Potassium (K⁺)
• Principal intracellular cation.
• Maintains intracellular fluid (ICF) volume and osmotic pressure.
• Resting membrane potential: High intracellular K⁺ and low extracellular K⁺ are critical for the resting potential of all
excitable cells.
• Repolarization in action potentials: K⁺ efflux restores the negative membrane potential after depolarization.
• Enzyme activation: K⁺ is a cofactor for many enzymes, especially in protein and carbohydrate metabolism.
• Acid-base balance: K⁺ is exchanged with H⁺ in the kidneys during acid-base regulation.
Chloride (Cl⁻)
• Principal extracellular anion.
• Maintains electroneutrality: Balances positive charges of Na⁺ and K⁺ in ECF and ICF.
• Osmotic balance: Contributes to ECF osmolarity.
• Acid-base balance: Involved in the chloride shift (exchange with bicarbonate in red blood cells) and gastric acid (HCl)
production in the stomach.
• Nerve function: Participates in inhibitory neurotransmission (GABA and glycine receptors).
Potassium, Sodium, and Blood Chlorides: Normal Levels

Ion Serum Reference Range

Sodium 135–145 mmol/L

Potassium 3.5–5.1 mmol/L

Chloride 98–107 mmol/L

Disorders: Hypo- and Hypernatremia, Hypo- and Hyperkalemia

Hyponatremia (↓ Na⁺ in blood)
• Definition: Serum Na⁺ < 135 mmol/L.
• Causes: Excess water intake, SIADH, diuretics, adrenal insufficiency, vomiting, diarrhea, heart/kidney/liver failure.
• Symptoms: Nausea, headache, confusion, seizures, coma (due to cerebral edema).
Hypernatremia (↑ Na⁺ in blood)
 • Definition: Serum Na⁺ > 145 mmol/L.
• Causes: Water loss (dehydration, diabetes insipidus), excessive Na⁺ intake.
• Symptoms: Thirst, weakness, irritability, confusion, seizures, coma (due to cellular dehydration).
Hypokalemia (↓ K⁺ in blood)
• Definition: Serum K⁺ < 3.5 mmol/L.
• Causes: Diuretics, vomiting, diarrhea, hyperaldosteronism, insulin excess.
• Symptoms: Muscle weakness, cramps, arrhythmias, paralysis, ECG changes (flattened T waves, U waves).
Hyperkalemia (↑ K⁺ in blood)
• Definition: Serum K⁺ > 5.1 mmol/L.
• Causes: Renal failure, hypoaldosteronism, tissue breakdown (hemolysis, rhabdomyolysis), acidosis, certain drugs.
• Symptoms: Muscle weakness, paresthesia, life-threatening cardiac arrhythmias (peaked T waves, widened QRS,
asystole).

131. Calcium, magnesium and phosphorus. Biological role. Regulation of calcium and phosphorus metabolism. The role and
mechanism of hormonal control. Involvement of vitamin D.
Ans. Biological Role
Calcium (Ca²⁺)
• Structural: Major component of bones and teeth (as hydroxyapatite).
• Cellular signaling: Essential for muscle contraction, neurotransmitter release, hormone secretion, and blood clotting.
• Enzyme cofactor: Activates many enzymes, including those in blood coagulation.
 • Membrane stability: Stabilizes cell membranes and affects permeability.
Phosphorus (as Phosphate, PO₄³⁻)
• Structural: With calcium, forms hydroxyapatite in bones and teeth.
• Energy metabolism: Component of ATP, GTP, creatine phosphate.
• Nucleic acids: Essential for DNA, RNA, and phospholipids in cell membranes.
• Buffering: Important in acid-base balance as part of the phosphate buffer system.
Magnesium (Mg²⁺)
• Enzyme cofactor: Required for >300 enzymes, especially those using ATP (e.g., kinases).
• Nucleic acid stability: Stabilizes DNA and RNA structures.
• Neuromuscular function: Modulates nerve impulse transmission and muscle contraction.
• Bone: ~60% of body Mg is in bone, contributing to structure.
Regulation of Calcium and Phosphorus Metabolism
Key Hormones:
• Parathyroid hormone (PTH)
• Calcitonin
• Vitamin D (Calcitriol, 1,25-dihydroxycholecalciferol)
Parathyroid Hormone (PTH)
• Source: Parathyroid glands.
• Stimulus: Low plasma Ca²⁺.
 • Actions:
• Bone: Stimulates osteoclasts → bone resorption → ↑ Ca²⁺ and phosphate release.
• Kidney: Increases Ca²⁺ reabsorption, decreases phosphate reabsorption (↑ Ca²⁺, ↓ PO₄³⁻ in blood).
• Vitamin D activation: Stimulates 1α-hydroxylase in kidney → ↑ active vitamin D (calcitriol) production.
• Net effect: ↑ plasma Ca²⁺, ↓ plasma phosphate.
Calcitonin
• Source: Thyroid gland (parafollicular C-cells).
• Stimulus: High plasma Ca²⁺.
• Actions:
• Bone: Inhibits osteoclast activity → ↓ bone resorption.
• Kidney: Promotes urinary excretion of Ca²⁺ and phosphate.
• Net effect: ↓ plasma Ca²⁺ (minor role in humans).
Vitamin D (Calcitriol)
• Source: Synthesized in skin (from 7-dehydrocholesterol via sunlight), activated in liver (25-hydroxylation) and kidney (1α-
hydroxylation).
• Actions:
• Intestine: Increases absorption of Ca²⁺ and phosphate.
• Bone: Stimulates bone resorption (with PTH), but also promotes bone mineralization by maintaining Ca²⁺ and
phosphate levels.
• Kidney: Increases reabsorption of Ca²⁺ and phosphate.
 • Net effect: ↑ plasma Ca²⁺ and phosphate.
Mechanism of Hormonal Control
• Negative feedback: Low Ca²⁺ → ↑ PTH → ↑ Ca²⁺ (restored); high Ca²⁺ → ↑ calcitonin → ↓ Ca²⁺.
• PTH and vitamin D act synergistically to increase plasma Ca²⁺, but have opposite effects on phosphate (PTH decreases,
vitamin D increases).
• Vitamin D synthesis is tightly regulated by PTH and plasma phosphate.

132. Methods for determination of serum calcium, diagnostic value.

Ans. Methods for Determination of Serum Calcium
1. Colorimetric Methods
• Principle: Calcium in serum reacts with specific chromogenic reagents to form a colored complex, the intensity of which
is measured spectrophotometrically.
• Common reagents:
• o-Cresolphthalein complexone (OCPC): Calcium forms a purple complex with OCPC in alkaline medium.
• Arsenazo III: Forms a blue-purple complex with calcium, suitable for automated analyzers.
• Procedure: Serum is mixed with the reagent, and absorbance is measured at a specific wavelength. The calcium
concentration is calculated by comparing to a standard curve.
2. Atomic Absorption Spectrophotometry (AAS)
• Principle: Measures the absorption of light by calcium atoms vaporized in a flame; highly specific and accurate.
• Use: Reference method in clinical laboratories, especially for research or when precise measurement is required.
3. Ion-Selective Electrode (ISE) Method
• Principle: Measures the activity of ionized (free) calcium directly using a calcium-selective electrode.
• Significance: Provides the physiologically active fraction of calcium, important in critical care settings.
4. Titrimetric Methods (Rarely Used Clinically)
• Principle: Calcium is titrated with a standard solution of a chelating agent (e.g., EDTA) using an indicator such as
murexide.
• Use: Mainly for research or in laboratories without automated equipment.
Diagnostic Value
• Normal Reference Range:
• Total serum calcium: 2.15–2.55 mmol/L (8.6–10.2 mg/dL)
• Ionized calcium: 1.12–1.32 mmol/L (4.5–5.3 mg/dL)
(Values may vary slightly by laboratory and population.)
• Clinical Significance:
• Hypercalcemia (↑ calcium): May indicate hyperparathyroidism, malignancy (bone metastases, multiple
myeloma), vitamin D intoxication, sarcoidosis, prolonged immobilization, or thiazide diuretic use.
• Hypocalcemia (↓ calcium): May result from hypoparathyroidism, vitamin D deficiency, chronic kidney disease,
acute pancreatitis, or massive transfusion (citrate binding).
• Ionized calcium: More accurately reflects the physiologically active calcium status, especially in critically ill
patients, those with abnormal plasma proteins, or acid-base disturbances.
• Diagnostic Use:
• Routine screening for metabolic, endocrine, renal, and bone disorders.
 • Monitoring of patients with parathyroid, kidney, or malignancy-related calcium disturbances.
• Evaluation of neuromuscular symptoms (tetany, muscle cramps, paresthesia), cardiac arrhythmias, or
unexplained bone disease.

133. Iron, sources, need, absorption, transport proteins, deposition, biological role.
Ans. Sources of Iron
• Dietary Sources:
• Heme iron: Found in animal products such as red meat, poultry, and fish; more efficiently absorbed.
• Non-heme iron: Found in plant foods like legumes, grains, nuts, leafy green vegetables, and fortified cereals; less
efficiently absorbed.
• Other factors:
• Vitamin C, gastric acid, and certain amino acids enhance non-heme iron absorption.
• Phytates (in grains), polyphenols (in tea/coffee), calcium, and some proteins can inhibit iron absorption64.
Need for Iron
• Recommended Dietary Allowance (RDA):
• Adult men: ~8–10 mg/day
• Adult women: ~18 mg/day (higher due to menstruation)
• Pregnant women: up to 27 mg/day
• Children and infants: higher per kg body weight due to growth
• Increased requirements:
 • Growth, pregnancy, lactation, menstruation, blood loss, chronic diseases6.
Absorption of Iron
• Site:
• Mainly in the duodenum and upper jejunum.
• Mechanism:
• Heme iron: Absorbed intact via heme transporter, then broken down in enterocytes to release Fe²⁺.
• Non-heme iron: Absorbed as Fe²⁺ after reduction from Fe³⁺ (by duodenal cytochrome b and vitamin C); taken up
by divalent metal transporter 1 (DMT1).
• Regulation:
• Absorption increases with body need and decreases with adequate stores.
• Hepcidin (a liver-derived hormone) inhibits iron absorption by degrading ferroportin, the iron exporter on
enterocytes.
Transport Proteins
• Transferrin:
• The main plasma protein that binds and transports Fe³⁺ in the blood to tissues, especially bone marrow for
erythropoiesis.
• Each transferrin molecule carries two Fe³⁺ ions.
• Other proteins:
• Ferroportin: Exports iron from enterocytes and macrophages into plasma.
• Lactoferrin: Binds iron in milk and neutrophil granules, with antimicrobial functions5.
 • Ceruloplasmin: Oxidizes Fe²⁺ to Fe³⁺, facilitating transferrin binding5.
Deposition (Storage) of Iron
• Ferritin:
• The primary intracellular iron storage protein, found in liver, spleen, bone marrow, and other tissues.
• Stores iron in a soluble, non-toxic form and releases it when needed.
• Hemosiderin:
• An insoluble, partially degraded form of ferritin, accumulates in conditions of iron overload.
• Total body iron stores:
• Adult: 3–4 g; about two-thirds in hemoglobin, the rest in storage and enzymes5.
Biological Role of Iron
• Oxygen transport and storage:
• Essential component of hemoglobin (in red blood cells) and myoglobin (in muscles), enabling oxygen transport
and storage5.
• Enzyme cofactor:
• Required for cytochromes (electron transport chain), catalase, peroxidase, and other enzymes involved in cellular
respiration and detoxification5.
• Cell growth and differentiation:
• Iron is critical for DNA synthesis and cell division.
• Immune function:
• Supports proliferation and maturation of immune cells.
 • Antioxidant defense:
• Involved in the activity of enzymes like catalase and peroxidase that neutralize reactive oxygen species.

134. Copper, sources, need, absorption. Transport in the bloodstream. Biological role.
Ans. Sources of Copper
• Dietary sources:
• Rich sources: Liver, shellfish (especially oysters), nuts, seeds (sunflower, sesame), legumes, whole grain products,
and dark chocolate.
• Other sources: Meat, fish, potatoes, green leafy vegetables, and dried fruits.
• Bioavailability:
• Animal-based copper is generally more bioavailable than plant-based copper due to less interference from
phytates and fiber.
Daily Need for Copper
• Recommended Dietary Allowance (RDA):
• Adults: 0.9 mg/day (900 µg/day)
• Children: 0.3–0.9 mg/day, depending on age
• Pregnancy/Lactation: Slightly increased needs
• Increased requirements:
• Periods of rapid growth (infancy, adolescence), pregnancy, and lactation.
Absorption of Copper
 • Site:
• Mainly absorbed in the small intestine (duodenum and upper jejunum).
• Mechanism:
• Copper is absorbed as Cu⁺ and Cu²⁺ ions, with the help of specific transporters.
• Enhancers: Amino acids and organic acids can increase absorption.
• Inhibitors: High intakes of zinc, iron, or phytates can decrease copper absorption.
• Efficiency:
• Normally, 30–60% of dietary copper is absorbed, but absorption increases during deficiency.
Transport in the Bloodstream
• After absorption:
• Copper enters the portal circulation, bound to albumin and amino acids.
• Hepatic processing:
• In the liver, copper is incorporated into ceruloplasmin (the main copper-carrying protein in plasma).
• Transport proteins:
• Ceruloplasmin: Carries about 90–95% of plasma copper; also functions as a ferroxidase, facilitating iron
metabolism.
• Albumin and transcuprein: Carry the remaining copper in plasma.
• Cellular uptake:
• Cells take up copper via specific transporters (e.g., CTR1).
Deposition and Storage
 • Liver: Main storage site; copper is stored bound to metallothionein and other proteins.
• Other tissues: Brain, heart, kidneys, and muscles also store copper in smaller amounts.
Biological Role of Copper
• Enzyme cofactor: Copper is essential for the activity of several key enzymes:
• Cytochrome c oxidase: Terminal enzyme in the mitochondrial electron transport chain (cellular respiration).
• Ceruloplasmin: Oxidizes Fe²⁺ to Fe³⁺, aiding iron transport by transferrin.
• Superoxide dismutase (SOD): Protects cells from oxidative damage (antioxidant defense).
• Lysyl oxidase: Required for collagen and elastin cross-linking (connective tissue strength).
• Tyrosinase: Involved in melanin synthesis (skin and hair pigmentation).
• Dopamine β-monooxygenase: Converts dopamine to norepinephrine (neurotransmitter synthesis).
• Iron metabolism: Copper is crucial for iron absorption, mobilization, and incorporation into hemoglobin.
• Immune function: Supports the development and maintenance of immune cells.
• Nervous system: Important for normal brain and nervous system development and function.
Clinical Notes
• Deficiency:
• Can lead to anemia (hypochromic, microcytic), neutropenia, bone abnormalities, impaired immune function, and
neurological symptoms.
• Menkes disease (genetic disorder) causes severe copper deficiency due to impaired absorption.
• Excess/Toxicity:
 • Rare, but can occur with genetic disorders (Wilson's disease-copper accumulation in liver, brain, eyes), or
excessive supplementation.

135. Trace elements: iodine, fluorine, manganese, specific functions.

Ans. Iodine
Biological Role:
• Essential for Thyroid Hormones: Iodine is a critical component of the thyroid hormones thyroxine (T₄) and
triiodothyronine (T₃).
• Regulation of Metabolism: These hormones control basal metabolic rate, influence protein synthesis, regulate growth
and development, and are essential for brain development and function.
• Mechanism: Iodine is actively taken up by the thyroid gland and incorporated into thyroglobulin, forming T₄ and T₃,
which are then released into the bloodstream5.
Deficiency Effects:
• Goiter: Enlargement of the thyroid gland.
• Hypothyroidism: Fatigue, weight gain, developmental delays in children.
• Cretinism: Severe deficiency during pregnancy can cause irreversible mental and physical retardation in the child5.
Fluorine (as Fluoride)
Biological Role:
• Tooth and Bone Health: Fluoride is incorporated into the hydroxyapatite crystals of teeth and bones, forming
fluoroapatite, which increases resistance to acid dissolution and dental caries (cavities).
• Enamel Strengthening: Reduces demineralization and enhances remineralization of tooth enamel.
 • Possible Role in Bone Mineralization: May help prevent osteoporosis at optimal intake levels.
Deficiency/Excess:
• Deficiency: Increased risk of dental caries.
• Excess (Fluorosis): Mottling of teeth, skeletal fluorosis (bone/joint changes).
Manganese
Biological Role:
• Enzyme Cofactor: Manganese is a cofactor for many enzymes, including:
• Superoxide dismutase (Mn-SOD): Antioxidant defense in mitochondria.
• Arginase: Urea cycle.
• Pyruvate carboxylase: Gluconeogenesis.
• Glycosyltransferases: Synthesis of glycoproteins and proteoglycans for cartilage and bone formation3.
• Bone and Connective Tissue Formation: Required for normal bone growth and cartilage synthesis.
• Metabolism: Involved in carbohydrate, amino acid, and cholesterol metabolism.
Deficiency/Excess:
• Deficiency: Rare, but can impair growth, bone formation, and reproductive function.
• Excess: Neurotoxicity (manganism) in occupational exposures.

136. Exogenous and endogenous water, sources, demand. The biological role of water.
Ans. Exogenous and Endogenous Water
Exogenous Water
• Definition: Water that enters the body from external sources.
• Main sources:
• Drinking water and beverages: The primary and most significant source.
• Water in food: Fruits, vegetables, soups, and other foods contain varying amounts of water.
• Contribution: Exogenous water generally accounts for the majority of daily water intake.
Endogenous Water
• Definition: Water produced within the body as a result of metabolic processes.
• Sources:
• Metabolic water: Produced during the oxidation of macronutrients (carbohydrates, fats, proteins) in cellular
respiration.
• For example, oxidation of 100 g of fat yields about 107 g of water; carbohydrates and proteins yield less.
• Contribution: Endogenous water forms a smaller, but physiologically important, portion of total water intake-especially
significant during fasting, hibernation, or in desert animals.
Sources and Daily Demand for Water
Sources of Water
1. Exogenous:
• Drinking water and other beverages.
• Water content in solid foods.
2. Endogenous:
 • Metabolic oxidation of nutrients.
Daily Water Requirement
• Average adult need: About 2–2.5 liters per day (can vary with age, activity, climate, health).
• Factors increasing demand: Physical activity, hot climate, fever, diarrhea, vomiting, lactation, pregnancy.
• Water losses: Occur via urine, feces, sweat, and exhaled air.
Biological Role of Water
• Universal solvent: Water dissolves electrolytes, nutrients, and waste products, enabling their transport and chemical
reactions.
• Medium for biochemical reactions: All cellular metabolism occurs in aqueous solutions.
• Thermoregulation: Water has a high heat capacity and is essential for maintaining body temperature via sweating and
evaporation.
• Transport: Water is the main component of blood and lymph, facilitating the distribution of nutrients, hormones, and
removal of wastes.
• Lubrication and protection: Water forms synovial fluid (joints), cerebrospinal fluid (brain/spinal cord), and is present in
mucus and serous fluids, protecting tissues and organs.
• Structural component: Water maintains cell turgor and tissue structure.
• Acid-base and osmotic balance: Water is vital for maintaining the correct osmotic pressure and pH in body fluids.

137. Regulation of water-salt metabolism. Structure, mechanism of action of vasopressin and aldosterone. Renin-angiotensin
system. Biochemical mechanisms of the development of renal hypertension.
Ans. Regulation of Water-Salt Metabolism
Water-salt (electrolyte) metabolism is crucial for maintaining blood volume, blood pressure, osmotic balance, and the proper
function of cells. The primary hormones involved are vasopressin (antidiuretic hormone, ADH) and aldosterone, with
regulation integrated by the renin-angiotensin-aldosterone system (RAAS).
Vasopressin (Antidiuretic Hormone, ADH)
Structure:
• Peptide hormone (nonapeptide) synthesized in the hypothalamus and released from the posterior pituitary.
Mechanism of Action:
• Target: Distal tubules and collecting ducts of the kidney.
• Receptor: Binds to V2 receptors on renal tubular cells, activating adenylate cyclase and increasing cAMP.
• Effect: Increases insertion of aquaporin-2 water channels into the apical membrane, enhancing water reabsorption.
• Physiological Result: Concentrates urine, conserves body water, and reduces plasma osmolarity.
Additional Effects:
• At higher concentrations, vasopressin causes vasoconstriction via V1 receptors, increasing blood pressure.
• Increases urea reabsorption, contributing to medullary hypertonicity and further water reabsorption1.
Regulation:
• Stimulated by: Increased plasma osmolarity, decreased blood volume/pressure.
• Inhibited by: Decreased plasma osmolarity, increased blood volume/pressure.
Aldosterone
Structure:
• Steroid hormone (mineralocorticoid) synthesized in the zona glomerulosa of the adrenal cortex.
Mechanism of Action:
• Target: Principal cells of the distal convoluted tubule and collecting duct in the kidney.
• Receptor: Intracellular mineralocorticoid receptor (acts as a transcription factor).
• Effect: Increases synthesis of epithelial sodium channels (ENaC) and Na⁺/K⁺-ATPase pumps.
• Physiological Result: Enhances sodium reabsorption and potassium excretion; water follows sodium osmotically,
expanding extracellular fluid volume and raising blood pressure2.
Regulation:
• Stimulated by: Angiotensin II (from RAAS), hyperkalemia (high plasma K⁺), ACTH (minor).
• Inhibited by: Atrial natriuretic peptide (ANP), high plasma sodium.
Renin-Angiotensin-Aldosterone System (RAAS)
Overview:
• A hormone cascade system that regulates blood pressure, fluid, and electrolyte balance.
Steps:
1. Renin release:
• Produced by juxtaglomerular cells in the kidney in response to low blood pressure, low Na⁺, or sympathetic
stimulation.
2. Angiotensinogen conversion:
• Renin cleaves liver-derived angiotensinogen to angiotensin I.
3. Angiotensin I to II:
• Angiotensin-converting enzyme (ACE, mainly in the lungs) converts angiotensin I to angiotensin II.
 4. Actions of Angiotensin II:
• Potent vasoconstrictor (raises systemic vascular resistance and BP).
• Stimulates aldosterone secretion from the adrenal cortex.
• Stimulates ADH release and thirst.
• Promotes sodium reabsorption in the proximal tubule.
Final effect:
• Increases blood volume and blood pressure by sodium and water retention, vasoconstriction, and increased thirst2.
Biochemical Mechanisms of Renal Hypertension
Renal (secondary) hypertension develops when the kidneys inappropriately activate mechanisms that increase blood
pressure, most commonly due to:
• Renal artery stenosis:
• Narrowing of the renal artery reduces renal perfusion pressure, stimulating renin release and activating RAAS,
leading to excessive vasoconstriction and sodium/water retention.
• Chronic kidney disease:
• Impaired sodium excretion increases blood volume and pressure.
• Excessive aldosterone production (Conn's syndrome):
• Leads to sodium retention, hypokalemia, and hypertension.
Biochemical features:
• Elevated plasma renin activity (in renovascular hypertension).
• Increased angiotensin II and aldosterone levels.
 • Enhanced sodium and water reabsorption, increased peripheral resistance.
• Persistent hypertension can cause further renal damage, perpetuating the cycle2.

138. Biochemistry of nervous tissue. Features of energy metabolism. Oxygen demand. Metabolism of carbohydrates, sources.
The role of glucose in the substrate and energy supply of the brain.
Ans. Features of Nervous Tissue Energy Metabolism
• High Metabolic Activity: Nervous tissue, especially the brain, is characterized by an exceptionally high metabolic rate.
Although the brain constitutes only 2–3% of body weight, it consumes about 20–25% of the body’s total oxygen at rest1.
• Intensive Blood Supply: This high metabolic demand is supported by an abundant blood supply, ensuring a constant
delivery of oxygen and nutrients1.
• Blood-Brain Barrier (BBB): The BBB restricts the entry of many substances, making the brain highly selective in its
substrate utilization. It allows glucose and certain amino acids to enter but largely excludes fatty acids and many other
metabolites1.
Oxygen Demand
• Very High Oxygen Consumption: Brain tissue has a respiration rate roughly 20 times higher than muscle tissue1.
• Dependence on Aerobic Metabolism: The brain is extremely sensitive to hypoxia; even brief interruptions in oxygen
supply can lead to loss of consciousness and, if prolonged, irreversible damage1.
• Reason: Most of the brain’s energy is required for maintaining ion gradients (Na⁺/K⁺-ATPase), generating action
potentials, synaptic transmission, and neurotransmitter cycling1.
Carbohydrate Metabolism in the Brain
Sources of Carbohydrates
 • Glucose: The principal and almost exclusive energy substrate for the adult brain under normal conditions1.
• Entry Mechanism: Glucose enters the brain via facilitated diffusion through the BBB using the GLUT1 transporter.
This process does not depend on insulin1.
• Glycogen Stores: The brain contains negligible glycogen (about 0.1% of brain mass), insufficient to support energy
needs for more than a few minutes1.
• Alternative Fuels: In prolonged fasting, starvation, or uncontrolled diabetes, the brain can adapt to use ketone bodies
(acetoacetate and β-hydroxybutyrate) as alternative energy sources1.
Pathways for Glucose Utilization
• Aerobic Glycolysis: Glucose is metabolized via glycolysis to pyruvate, which enters the mitochondria and is oxidized to
CO₂ and H₂O in the tricarboxylic acid (TCA) cycle, generating ATP13.
• Pentose Phosphate Pathway: Provides NADPH for reductive biosynthesis and antioxidant defense, and ribose-5-
phosphate for nucleotide synthesis13.
• Biosynthetic Roles: Intermediates from glucose metabolism are used for the synthesis of neurotransmitters (e.g.,
glutamate, GABA), nucleotides, and other essential molecules13.
Role of Glucose in the Substrate and Energy Supply of the Brain
• Primary Energy Source: Glucose is the main-and under normal conditions, the only-energy substrate for neurons and
glial cells13.
• Continuous Supply Needed: Because the brain cannot store significant amounts of glucose or glycogen, it is entirely
dependent on a constant supply from the bloodstream1.
• Insulin Independence: Glucose uptake into the brain is independent of insulin, unlike in muscle and adipose tissue1.
• Energy for Neuronal Activity: Most of the energy derived from glucose is used to maintain membrane potentials,
support synaptic transmission, and fuel the synthesis and recycling of neurotransmitters1.
 • Clinical Relevance: Hypoglycemia (low blood glucose) rapidly impairs brain function, leading to confusion, seizures,
coma, and potentially death if not corrected1.

139. The chemical composition of the nervous tissue. Proteins, functional classification. Neurospecific proteins of the nervous
tissue.
Ans. Chemical Composition of Nervous Tissue
Nervous tissue is composed of:
• Water: 68–83% (higher in gray matter)
• Proteins: 8–9%
• Lipids: 5–17% (notably high in white matter due to myelin)
• Minerals: 1–2%
Proteins are a major component of both gray and white matter, playing structural, enzymatic, transport, and signaling roles5.
Functional Classification of Proteins in Nervous Tissue
Nervous tissue proteins can be functionally classified as follows:
1. Structural Proteins
• Cytoskeletal proteins: Neurofilaments, microtubule-associated proteins (MAPs, including tau protein), tubulin,
actin.
• Myelin proteins: Myelin basic protein (MBP), proteolipid protein (PLP), myelin-associated glycoprotein (MAG).
• Synaptic proteins: Synapsins, synaptophysin, PSD-95.
2. Enzymatic Proteins
 • Enzymes for neurotransmitter synthesis and degradation: Choline acetyltransferase, acetylcholinesterase,
tyrosine hydroxylase, monoamine oxidase, glutamate decarboxylase.
• Metabolic enzymes: Hexokinase, phosphofructokinase, isocitrate dehydrogenase (for energy metabolism).
3. Transport Proteins
• Ion channels and pumps: Na⁺/K⁺-ATPase, Ca²⁺-ATPase, voltage-gated sodium, potassium, and calcium channels.
• Neurotransmitter transporters: e.g., dopamine transporter, serotonin transporter.
4. Receptor Proteins
• Neurotransmitter receptors: NMDA, AMPA, GABA_A, nicotinic and muscarinic acetylcholine receptors, dopamine,
serotonin, and other monoamine receptors.
5. Neurospecific Proteins
• Neuron-specific enolase (NSE): A glycolytic enzyme used as a clinical marker for neuronal injury.
• S100 proteins: Calcium-binding proteins found in glial cells, used as markers for glial activity or damage.
• Glial fibrillary acidic protein (GFAP): An intermediate filament protein specific to astrocytes, used as a marker for
astroglial cells.
• Synaptophysin: A synaptic vesicle protein, marker for synapses.
• Neurofilament proteins: Provide structural support for axons; used as markers for neuronal integrity.
• Tau protein: Microtubule-associated protein important in axonal transport and implicated in neurodegenerative
diseases.
Neurospecific Proteins of Nervous Tissue
Key neurospecific proteins include:
• Neuron-Specific Enolase (NSE):
• Found in neurons and neuroendocrine cells.
• Used clinically as a marker for neuronal damage and some tumors5.
• Glial Fibrillary Acidic Protein (GFAP):
• Specific to astrocytes (a type of glial cell).
• Marker for astrocyte activity and gliosis.
• Myelin Basic Protein (MBP):
• Major component of myelin sheath, specific for oligodendrocytes and Schwann cells.
• Marker for demyelinating diseases.
• S100 Proteins:
• Family of calcium-binding proteins, mainly in glial cells.
• S100B is used as a marker for glial activation or injury.
• Synaptophysin:
• Integral membrane glycoprotein of synaptic vesicles.
• Marker for synaptic density.
• Neurofilament Proteins:
• Intermediate filament proteins unique to neurons.
• Used as markers for axonal damage.
• Tau Protein:
 • Associated with microtubules in neurons; abnormal phosphorylation is linked to neurodegenerative diseases (e.g.,
Alzheimer’s).

140. Lipids and carbohydrates of the brain: representatives, biological role. Features of the exchange.
Ans. Carbohydrates of the Brain
Main Representatives:
• Glucose: The principal and almost exclusive carbohydrate used by the adult brain for energy.
• Glycogen: Present in very small amounts (~0.1% of brain mass), mainly in astrocytes.
Biological Role:
• Primary Energy Source: Glucose is the main substrate for ATP production in neurons and glial cells under normal
conditions.
• Neurotransmitter Synthesis: Glucose metabolism provides precursors for neurotransmitter biosynthesis (e.g.,
glutamate, GABA, acetylcholine).
• Pentose Phosphate Pathway: Supplies NADPH for antioxidant defense and ribose-5-phosphate for nucleotide synthesis.
Metabolic Features:
• High Demand: The brain consumes 20–25% of total body oxygen and a proportional amount of glucose, despite
representing only 2–3% of body mass.
• Dependence on Continuous Supply: The brain stores little glycogen and relies on a constant supply of glucose from the
blood, transported via GLUT1 (insulin-independent).
• Limited Use of Other Fuels: Fatty acids do not cross the blood-brain barrier and are not used as an energy source.
During prolonged fasting or diabetes, the brain can adapt to use ketone bodies.
 • High Hexokinase Activity: Ensures rapid phosphorylation and utilization of glucose1.
Lipids of the Brain
Main Representatives:
• Phospholipids: Phosphatidylserine, phosphatidylcholine, phosphatidylethanolamine, phosphatidylinositol,
plasmalogens.
• Sphingolipids: Sphingomyelin (major in myelin), cerebrosides, sulfatides, gangliosides.
• Cholesterol: High concentration, especially in myelin.
• Glycolipids: Cerebrosides (white matter/myelin), gangliosides (neuronal membranes).
Biological Role:
• Structural: Lipids are essential for the formation and maintenance of neuronal and glial cell membranes, and especially
for the multilayered myelin sheath that insulates axons and enables rapid nerve impulse conduction.
• Electrical Insulation: Myelin lipids provide dielectric properties, crucial for saltatory conduction.
• Signal Transduction: Membrane lipids (especially phospholipids and gangliosides) participate in cell signaling and
synaptic transmission.
• Antioxidant Protection: Gangliosides act as antioxidants and protect against lipid peroxidation; they also promote
healing after brain injury.
• Development: Synthesis of cerebrosides and gangliosides is especially high during brain development and myelination.
Metabolic Features:
• High Lipid Content: The brain is exceptionally rich in lipids (5–17% of tissue mass), with white matter (myelin) being
particularly lipid-dense.
 • Unique Lipid Composition: The brain contains little to no neutral fats (triacylglycerols); cholesterol esters are mainly
present during active myelination.
• Slow Lipid Turnover: Most brain lipids are renewed slowly, except during development.
• Cholesterol Synthesis: Active in the developing brain; in adults, synthesis is low.
• Restricted Fatty Acid Use: Free fatty acid content is very low; the brain relies on structural, not energy, roles for lipids1.

141. Free amino acid fund in the brain. Glutamic acid metabolic pathways. Formation of GABA, a role in the brain.
Ans. Free Amino Acid Pool in the Brain
• High Concentration: The concentration of free amino acids in the brain is about 8 times higher than in blood, due to
active transport systems for various amino acid types across the blood-brain barrier and intensive metabolic exchange
with the blood1.
• Dominant Amino Acids: Glutamate and aspartate, along with their derivatives (glutamine, GABA, acetyl derivatives),
make up approximately 70–75% of the total free amino acids in nervous tissue1.
• Functional Importance: These amino acids are not only building blocks for protein synthesis but also serve as key
neurotransmitters and metabolic intermediates.
Glutamic Acid (Glutamate) Metabolic Pathways in the Brain
Glutamic acid (glutamate) is central to brain metabolism and neurotransmission:
Main Pathways:
1. Transamination:
• Glutamate is rapidly formed from α-ketoglutarate (an intermediate of the TCA cycle) by transamination reactions.
 • This links amino acid metabolism with the TCA cycle, allowing for the exchange of nitrogen and carbon
skeletons15.
2. Glutamine Synthesis:
• Glutamate can be converted to glutamine by glutamine synthetase in astrocytes, serving as a temporary
neutralization of ammonia and a reservoir for neurotransmitter cycling1.
3. GABA Synthesis (GABA Shunt):
• Glutamate is decarboxylated by the enzyme glutamate decarboxylase (GAD, requires vitamin B6/PLP) to form γ-
aminobutyric acid (GABA), the major inhibitory neurotransmitter in the CNS15.
4. Glutathione Synthesis:
• Glutamate is a precursor for glutathione, a critical antioxidant in the brain1.
5. Energy Metabolism:
• Glutamate can be deaminated to α-ketoglutarate, entering the TCA cycle for energy production or anaplerosis15.
Formation of GABA and Its Role in the Brain
GABA Synthesis:
• Enzyme: Glutamate decarboxylase (GAD)
• Reaction:
Glutamate→GAD (PLP)GABA+CO2GlutamateGAD (PLP)GABA+CO2
• Localization: Occurs within GABAergic neurons; GABA cannot cross the blood-brain barrier and must be synthesized in
situ15.
GABA Shunt:
 • GABA can be transaminated to succinic semialdehyde, then oxidized to succinate, which enters the TCA cycle-a bypass
known as the "GABA shunt"5.
Role of GABA in the Brain:
• Major Inhibitory Neurotransmitter: GABA is the principal inhibitory neurotransmitter in the cerebral cortex and much of
the CNS15.
• Function: It reduces neuronal excitability by increasing chloride ion influx through GABA_A receptors or activating
GABA_B metabotropic receptors, leading to hyperpolarization and inhibition of action potentials.
• Balance with Glutamate: The balance between excitatory (glutamate) and inhibitory (GABA) neurotransmission is
crucial for normal brain function, affecting processes such as motor control, anxiety, sleep, and seizure susceptibility.
• Clinical Relevance: Altered GABAergic transmission is implicated in epilepsy, anxiety disorders, and other
neuropsychiatric conditions.

142. Biogenic amines, mechanism of formation, functions in the brain, inactivation.

Ans.
143. Biologically active peptides of nervous tissue. Role in the perception of pain and pain relief, in the regulation of autonomic
and higher functions of the nervous system.
Ans. Biologically active peptides in the nervous system-often called neuropeptides-are short chains of amino acids that
function as neurotransmitters, neuromodulators, or neurohormones. They play crucial roles in the perception and modulation
of pain, pain relief, and the regulation of both autonomic (vegetative) and higher (cognitive, emotional) nervous system
functions2.
Key Neuropeptides and Their Functions
1. Opiate-Like Peptides (Endorphins and Enkephalins)
 • Endorphins (e.g., β-endorphin) and enkephalins (methionine- and leucine-enkephalin) are produced in the brain and
spinal cord.
• Role in Pain and Pain Relief:
• Bind to opioid receptors, mimicking the effects of opiate drugs like morphine.
• Inhibit the transmission of pain signals in the central nervous system, producing analgesia (pain relief) and
euphoria.
• Released during stress, exercise, and certain emotional states, contributing to the “runner’s high” and natural pain
suppression2.
2. Substance P
• Function:
• A key neurotransmitter in the transmission of pain signals from peripheral nerves to the central nervous system.
• Involved in neurogenic inflammation and the regulation of mood and anxiety.
• Pain Perception:
• Promotes pain sensation (nociception) and is released in response to painful stimuli.
3. Somatostatin
• Functions:
• Acts as an inhibitory neuromodulator in the brain.
• Inhibits the release of many other neurotransmitters and neuropeptides.
• Modulates pain perception and is involved in the regulation of endocrine and autonomic functions4.
4. Neurohypophyseal Peptides (Vasopressin and Oxytocin)
 • Vasopressin (antidiuretic hormone, ADH):
• Involved in water balance, blood pressure regulation, and social behaviors.
• Modulates memory and stress responses.
• Oxytocin:
• Regulates social bonding, sexual behavior, childbirth, and lactation.
• Influences emotional and social behaviors, including trust and empathy2.
5. Other Neuropeptides
• Cholecystokinin (CCK):
• Present in the brain and gut; involved in satiety, anxiety, and pain modulation.
• Gastrin, Secretin, Vasoactive Intestinal Peptide (VIP):
• Modulate gastrointestinal function, but also act as neuromodulators in the brain, influencing circadian rhythms,
learning, and memory.
• Sleep Peptides:
• Some peptides induce sleep when administered experimentally, though their precise molecular nature is still
being studied2.
Role in the Perception and Modulation of Pain
• Pain Transmission:
• Substance P and related peptides facilitate the transmission of pain signals in the spinal cord and brain.
• Pain Inhibition (Analgesia):
 • Endorphins, enkephalins, and dynorphins inhibit pain pathways by activating opioid receptors, reducing
neurotransmitter release, and hyperpolarizing neurons.
• This endogenous opioid system is the body’s natural mechanism for pain control2.
Regulation of Autonomic and Higher Nervous Functions
• Autonomic Regulation:
• Neuropeptides such as somatostatin, VIP, and neuropeptide Y regulate heart rate, blood pressure, gastrointestinal
motility, and secretion.
• They modulate the activity of the sympathetic and parasympathetic nervous systems.
• Higher Functions:
• Peptides like oxytocin and vasopressin influence social behavior, emotional responses, memory, and learning.
• Some peptides (e.g., sleep peptides, memory peptides) are involved in sleep-wake cycles and memory
formation2.

144. Biochemistry of muscle tissue. Contractile and regulatory proteins of muscles: myosin, antin, actomyosin, tropomyosin,
troponin.
Ans. Overview of Muscle Tissue Biochemistry
Muscle tissue is specialized for contraction, enabling movement, posture maintenance, and vital physiological processes (e.g.,
heart beating, peristalsis). Its unique biochemical properties arise from a highly organized system of contractile and regulatory
proteins within the myofibrils of muscle fibers.
Main Contractile Proteins
1. Myosin
 • Structure: Large, fibrous protein with a long tail and globular head region; composed of heavy and light chains.
• Function: ATPase activity in the head region hydrolyzes ATP, providing energy for contraction. Myosin heads bind to actin
filaments and, through conformational changes, generate the force for muscle contraction.
2. Actin
• Structure: Exists as globular monomers (G-actin) that polymerize to form filamentous actin (F-actin).
• Function: Forms the backbone of the thin filaments. Provides binding sites for myosin heads during contraction.
3. Actomyosin
• Definition: A complex formed by the interaction of actin and myosin during muscle contraction.
• Function: The cyclic formation and breaking of actomyosin cross-bridges underlie the sliding filament mechanism of
muscle contraction.
Main Regulatory Proteins
4. Tropomyosin
• Structure: Long, double-helical protein that runs along the groove of the F-actin filament.
• Function: In resting muscle, tropomyosin blocks the myosin-binding sites on actin, preventing contraction.
5. Troponin
• Structure: A complex of three subunits:
• Troponin C (TnC): Binds calcium ions.
• Troponin I (TnI): Inhibits actin-myosin interaction.
• Troponin T (TnT): Binds to tropomyosin, anchoring the troponin complex.
 • Function: Upon binding of Ca²⁺ to TnC (during excitation), troponin undergoes a conformational change that shifts
tropomyosin away from the myosin-binding sites on actin, allowing contraction to proceed.
Mechanism of Muscle Contraction: The Sliding Filament Theory
1. Resting State:
• Tropomyosin covers myosin-binding sites on actin; troponin complex holds tropomyosin in place.
2. Excitation:
• Nerve impulse triggers Ca²⁺ release from the sarcoplasmic reticulum.
• Ca²⁺ binds to troponin C, causing a conformational change.
• Tropomyosin shifts, exposing binding sites on actin.
3. Cross-Bridge Formation:
• Myosin heads bind to exposed sites on actin, forming actomyosin cross-bridges.
4. Power Stroke:
• Myosin heads pivot, pulling actin filaments toward the center of the sarcomere (muscle contraction).
• ATP binds to myosin, causing detachment from actin and resetting the head for another cycle.

145. Sarcoplasmic proteins. Myoglobin, structure, function.

Ans. Sarcoplasmic Proteins
• Definition:
Sarcoplasmic proteins are a group of water-soluble proteins found in the sarcoplasm (the cytoplasm) of muscle cells.
They comprise about 30% of the total muscle proteins and are easily extracted with neutral salt solutions.
 • Types:
These include enzymes involved in glycolysis and other metabolic pathways, myoglobin, and various regulatory proteins.
• Functions:
• Participate in muscle metabolism (energy production, glycolysis, etc.)
• Serve as oxygen reservoirs (myoglobin)
• Play roles in ion transport and cellular signaling
Myoglobin: Structure
• Classification:
Myoglobin is a chromoprotein and a member of the hemoprotein family, similar to hemoglobin but functionally and
structurally distinct17.
• Molecular Structure:
• Globular protein consisting of a single polypeptide chain (~153 amino acids).
• Contains one heme group (prosthetic group) embedded in a hydrophobic pocket.
• The heme consists of protoporphyrin IX with a central iron atom (Fe²⁺) that can reversibly bind one molecule of
oxygen.
• The polypeptide (apoprotein) is called globin; together with heme, it forms the holoprotein myoglobin17.
• The 3D structure is predominantly α-helical (about 75% of residues), with eight α-helices labeled A–H4.
• Comparison to Hemoglobin:
• Myoglobin is a monomer (single subunit), while hemoglobin is a tetramer (four subunits).
• Myoglobin binds one O₂ molecule; hemoglobin binds four.
Myoglobin: Function
• Oxygen Storage:
Myoglobin serves as an intracellular reservoir of oxygen in skeletal and cardiac muscle, facilitating oxygen supply during
periods of high demand or low oxygen availability17.
• Oxygen Transport:
• Increases the rate of oxygen diffusion from the cell membrane to the mitochondria in muscle fibers.
• Releases oxygen when the partial pressure of O₂ in muscle falls (e.g., during intense exercise or hypoxia).
• Protective Role:
• Accumulates oxygen, protecting muscle cells from temporary oxygen starvation.
• Color of Muscle:
• Myoglobin gives muscle its characteristic red color due to the heme group17.
• Clinical Significance:
• Myoglobin is released into the blood after muscle injury (e.g., myocardial infarction), and its detection in
plasma/urine is used as a marker for muscle damage.

146. Biochemical mechanisms of muscle contraction and relaxation. The role of regulatory proteins, calcium.
Ans. 1. Overview of Muscle Contraction
Muscle contraction is a highly coordinated biochemical process involving the interaction of contractile proteins (actin and
myosin), regulatory proteins (tropomyosin and troponin), and the essential signaling role of calcium ions (Ca²⁺). The process is
powered by ATP and is reversible, allowing for both contraction and relaxation.
2. Sequence of Events in Muscle Contraction
A. Resting State
• Tropomyosin covers the myosin-binding sites on actin filaments, preventing interaction between actin and myosin.
• Troponin complex (composed of TnC, TnI, and TnT subunits) is attached to tropomyosin and actin, stabilizing this
blocked state.
B. Initiation: Calcium Release
• A nerve impulse triggers the release of Ca²⁺ from the sarcoplasmic reticulum (SR) into the sarcoplasm (muscle cell
cytoplasm).
• Increase in cytosolic Ca²⁺ is the key signal for contraction.
C. Role of Regulatory Proteins
• Calcium binds to Troponin C (TnC), causing a conformational change in the troponin complex.
• This shift moves tropomyosin away from the myosin-binding sites on actin.
D. Cross-Bridge Formation
• Myosin heads (with bound ADP and Pi) bind to the now-exposed sites on actin, forming cross-bridges.
• Power stroke: Myosin heads pivot, pulling actin filaments toward the sarcomere center; ADP and Pi are released.
• ATP binding: New ATP binds to myosin, causing it to detach from actin.
• ATP hydrolysis: Myosin hydrolyzes ATP to ADP + Pi, re-cocking the head for another cycle.
E. Cycle Repeats
• As long as Ca²⁺ and ATP are present, the cross-bridge cycle continues, resulting in muscle shortening (contraction).
3. Muscle Relaxation
 • Calcium Removal:
Ca²⁺ is actively pumped back into the sarcoplasmic reticulum by the Ca²⁺-ATPase pump (SERCA).
• Troponin-Tropomyosin Reset:
As cytosolic Ca²⁺ falls, Ca²⁺ dissociates from TnC, causing tropomyosin to return to its position blocking the myosin-
binding sites on actin.
• Cessation of Cross-Bridge Cycling:
Myosin can no longer bind to actin, and the muscle fiber relaxes.
5. The Central Role of Calcium
• Signal for Contraction:
Ca²⁺ is the essential trigger that initiates muscle contraction by binding to troponin C.
• Regulation:
The rapid release and reuptake of Ca²⁺ by the sarcoplasmic reticulum precisely control the timing and duration of
contraction and relaxation.
• ATP Dependence:
Both contraction (cross-bridge cycling) and relaxation (Ca²⁺ reuptake) require ATP.
6. Clinical Relevance
• Disorders of calcium handling or regulatory proteins (e.g., mutations in troponin or tropomyosin) can lead to muscle
weakness, cardiomyopathies, or abnormal muscle contraction.
• Measurement of troponin levels is used clinically as a marker of cardiac muscle injury.

147. Features of the energy metabolism of muscle tissue, creatiphosphate.

Ans. Features of Muscle Energy Metabolism
Muscle tissue is uniquely adapted for rapid, high-demand energy production and utilization, supporting both short bursts of
intense activity and sustained contraction. Key features include:
1. Multiple Energy Systems
• ATP: The immediate energy source for muscle contraction. However, ATP stores in muscle are very limited and can
sustain maximal contraction for only a few seconds.
• Phosphagen System (Creatine Phosphate): Acts as a rapid buffer and reservoir for ATP regeneration.
• Anaerobic Glycolysis: Provides ATP quickly by breaking down glucose to lactic acid, especially important during intense,
short-term activity when oxygen is limited.
• Aerobic Metabolism: During prolonged, less intense activity, muscles utilize aerobic oxidation of glucose, fatty acids, and
(to a lesser extent) amino acids in mitochondria, generating much larger amounts of ATP.
2. Glycogen Storage and Utilization
• Glycogen: Muscles store glycogen as a readily available source of glucose for glycolysis, especially during exercise.
• Glycogenolysis: Rapidly mobilizes glucose-1-phosphate for glycolysis during increased energy demand6.
3. Adaptability to Oxygen Availability
• Anaerobic Conditions: When oxygen is scarce (e.g., during intense exercise), muscles rely on glycolysis, producing
lactate as an end product. This allows ATP generation to continue, but less efficiently.
• Aerobic Conditions: With sufficient oxygen, pyruvate from glycolysis enters the mitochondria for complete oxidation via
the TCA cycle and oxidative phosphorylation, yielding much more ATP per glucose molecule2.
4. Fatty Acid Oxidation
• Rest and Prolonged Exercise: Muscles can oxidize fatty acids for ATP production, especially during prolonged, moderate-
intensity activity4.
Creatine Phosphate (Phosphocreatine) System
Role and Mechanism
• Creatine phosphate (CP) is a high-energy phosphate compound present in muscle cells.
• Function: Serves as a rapid, short-term energy buffer to regenerate ATP from ADP during the initial seconds of muscle
contraction.
• Reaction (catalyzed by creatine kinase):
Creatine phosphate+ADP↔Creatine+ATPCreatine phosphate+ADP↔Creatine+ATP
• Significance: This reaction allows muscles to maintain ATP levels during sudden, intense activity (e.g., sprinting, lifting),
even before glycolysis or oxidative phosphorylation can ramp up.
Features
• Speed: The creatine phosphate system provides energy almost instantaneously.
• Capacity: CP stores are limited and can sustain maximal muscle contraction for only about 5–8 seconds.
• Recovery: During rest, ATP produced by oxidative metabolism is used to regenerate creatine phosphate from creatine.
Clinical and Diagnostic Relevance
• Creatine kinase (CK): The enzyme catalyzing this reaction is a key marker for muscle damage (e.g., myocardial infarction,
rhabdomyolysis).

148. Extractive substances of muscles, creatine, creatinuria.

Ans. Extractive Substances of Muscles
Extractive substances are low-molecular-weight, water-soluble compounds found in muscle tissue. They are not structural
proteins or enzymes but are important for muscle metabolism and function. Major extractive substances include:
• Creatine and creatine phosphate (phosphocreatine)
• Creatinine
• Free amino acids
• Urea and uric acid
• Inosine, hypoxanthine, xanthine (purine derivatives)
• Lactic acid
• Other metabolic intermediates
These substances play roles in energy storage and transfer, nitrogen metabolism, and as intermediates or end-products of
muscle metabolism.
Creatine: Structure, Function, and Metabolism
Structure and Sources
• Creatine is a nitrogen-containing compound synthesized mainly in the liver, kidneys, and pancreas from the amino acids
glycine, arginine, and methionine.
• It is transported via the bloodstream to muscle tissue, where the majority (about 95%) is stored.
Function in Muscle
• Energy Buffer: In muscle cells, creatine is phosphorylated to form creatine phosphate (phosphocreatine) by the enzyme
creatine kinase.
• ATP Regeneration: During muscle contraction, phosphocreatine donates its phosphate group to ADP to rapidly
regenerate ATP, the immediate energy source for muscle contraction:
Creatine phosphate+ADP→creatine kinaseCreatine+ATPCreatine phosphate+ADPcreatine kinaseCreatine+ATP
• This system provides a quick energy reserve for short bursts of high-intensity muscle activity.
Metabolic Fate
• Non-enzymatic Conversion: Creatine and phosphocreatine are spontaneously and irreversibly converted to creatinine,
which is then excreted in the urine.
Creatinuria
Definition
• Creatinuria is the presence of an abnormally high concentration of creatine in the urine.
Causes
• Physiological: In children, pregnant women, and after intense muscle activity, some creatinuria may be observed due to
increased muscle metabolism.
• Pathological: Persistent creatinuria in adults may indicate:
• Muscle diseases (e.g., muscular dystrophy, myopathies)
• Hyperthyroidism
• Starvation or severe protein deficiency
• Impaired utilization of creatine by muscles

149. Connective tissue. The chemical composition of the connective tissue. The structure and structure of collagen and elastin,
properties, biological role. The role of vitamin C in biosynthesis.
Ans. 1. Chemical Composition of Connective Tissue
Connective tissue is a diverse group of tissues that support, protect, and give structure to other tissues and organs. Its main
components are:
• Cells:
• Fibroblasts (main producers of matrix components), chondrocytes, osteocytes, adipocytes, macrophages, mast
cells, and leukocytes.
• Extracellular Matrix (ECM):
• Fibrous proteins: Collagen, elastin, reticulin.
• Proteoglycans and glycosaminoglycans (GAGs): Hyaluronic acid, chondroitin sulfate, keratan sulfate, dermatan
sulfate, heparan sulfate.
• Glycoproteins: Fibronectin, laminin.
The ECM dominates the volume of connective tissue, with relatively few cells1.
2. Structure and Properties of Collagen
Collagen Structure
• Most abundant protein in mammals; main ECM protein in connective tissue.
• Types:
• Type I: Skin, tendon, bone, organs (major type).
• Type II: Cartilage.
• Type III: Reticular fibers.
• Type IV: Basal lamina.
• Molecular Structure:
 • Composed of three polypeptide α-chains forming a right-handed triple helix (“superhelix”).
• High glycine (every third residue), proline, and hydroxyproline content.
• Unique secondary structure: Gly-X-Y repeat, where X and Y are often proline or hydroxyproline.
• Levels of Organization:
• α-chain → triple helix → fibrils → fibers1.
Properties and Biological Role
• High tensile strength: Provides structural integrity to skin, bone, tendons, cartilage, and blood vessels.
• Resistance to stretching: Essential for mechanical support.
• Tissue repair: Collagen synthesis increases during wound healing.
• Biological role:
• Supports and connects tissues, acts as a scaffold for cell attachment, and regulates cell behavior via ECM
signaling1.
3. Structure and Properties of Elastin
Elastin Structure
• Major protein of elastic fibers in ECM.
• Produced by: Fibroblasts, smooth muscle cells, chondrocytes.
• Structure:
• Composed of tropoelastin monomers, cross-linked via desmosine and isodesmosine residues.
• Highly hydrophobic, rich in glycine, alanine, valine, and proline.
• Organization:
 • Forms a random coil structure, allowing for stretch and recoil1.
Properties and Biological Role
• Elasticity: Allows tissues (e.g., skin, lungs, blood vessels) to stretch and return to original shape.
• Resilience: Critical for normal function of arteries (withstand pulsatile blood flow), lungs (expand/contract during
breathing), and skin1.
4. Biosynthesis of Collagen and the Role of Vitamin C
Collagen Biosynthesis Steps
1. Synthesis of prepro-α chains in fibroblasts (intracellular).
2. Post-translational modifications:
• Hydroxylation of proline and lysine residues by prolyl and lysyl hydroxylases (requires vitamin C, Fe²⁺, O₂).
• Glycosylation of hydroxylysine residues.
3. Triple helix formation: Procollagen is formed and secreted.
4. Extracellular processing:
• Propeptides are cleaved to form tropocollagen.
• Collagen molecules assemble into fibrils and are stabilized by covalent cross-links (lysyl oxidase).
5. Maturation: Formation of collagen fibers1.
Role of Vitamin C (Ascorbic Acid)
• Essential cofactor for prolyl and lysyl hydroxylases.
• Function:
• Hydroxylation of proline and lysine is critical for stable triple helix formation and cross-linking of collagen fibers.
 • Deficiency (Scurvy):
• Impaired collagen synthesis leads to defective connective tissue, resulting in fragile blood vessels, poor wound
healing, bleeding gums, and bone abnormalities34.

150. Collagen. Chemical structure, biosynthesis, peculiarities of amino acid composition, posttranslational process. The
biological role of collagen. Collagen metabolism markers. Collagenase. Role in wound healing.
Ans. Chemical Structure of Collagen
• Collagen is the most abundant protein in mammals and the main structural protein of the extracellular matrix in
connective tissues1.
• Molecular Structure:
• Collagen consists of three polypeptide α-chains wound together in a right-handed triple helix (superhelix)1.
• Amino Acid Sequence: The repeating sequence is Gly-X-Y, where X is often proline and Y is often hydroxyproline1.
• Amino Acid Composition:
• Glycine: ~33% (every third residue).
• Proline and Hydroxyproline: ~15% combined (very high compared to other proteins).
• Hydroxylysine: Present, important for cross-linking.
• Low/absent: Tryptophan, cysteine; low tyrosine, methionine1.
• Types:
• Type I: Skin, tendon, bone, organs (most abundant).
 • Type II: Cartilage.
• Type III: Reticular fibers.
• Type IV: Basal lamina1.
Biosynthesis of Collagen
1. Synthesis of Preprocollagen (Intracellular):
• Collagen is synthesized as prepro-α-chains in fibroblasts (or chondroblasts, osteoblasts, etc.)1.
• Signal peptides are removed after entry into the rough endoplasmic reticulum (RER).
2. Posttranslational Modifications (Intracellular):
• Hydroxylation:
• Specific proline and lysine residues are hydroxylated to hydroxyproline and hydroxylysine by prolyl and lysyl
hydroxylases.
• Vitamin C (ascorbate) is an essential cofactor for these enzymes12.
• Iron (Fe²⁺) and O₂ are also required.
• Glycosylation:
• Some hydroxylysine residues are glycosylated with glucose or galactose.
• Triple Helix Formation:
• Three modified α-chains align and form a triple helix, stabilized by hydrogen bonds.
3. Secretion and Extracellular Processing:
• Procollagen is secreted into the extracellular space.
• Cleavage:
 • Propeptides at both ends are cleaved by specific peptidases, forming tropocollagen.
• Fibril Assembly:
• Tropocollagen molecules spontaneously assemble into collagen fibrils.
• Cross-linking:
• Lysyl oxidase (copper-dependent) forms covalent cross-links between lysine/hydroxylysine residues, stabilizing
fibrils.
Peculiarities of Amino Acid Composition
• High Glycine Content: Every third residue, allowing tight packing of the triple helix1.
• High Proline and Hydroxyproline: Impart rigidity and thermal stability.
• Hydroxylysine: Unique to collagen, important for glycosylation and cross-linking.
• Absence/Low Content: Tryptophan, cysteine, tyrosine, methionine1.
Biological Role of Collagen
• Structural Support: Main component of skin, bone, cartilage, tendons, ligaments, and blood vessels1.
• Tissue Integrity: Provides tensile strength and resistance to stretching.
• Wound Healing: Forms the scaffold for tissue repair and regeneration.
• Cellular Functions: Influences cell adhesion, migration, differentiation, and tissue morphogenesis.
• Basement Membranes: Type IV collagen is crucial for filtration barriers (e.g., in kidneys)1.
Collagen Metabolism Markers
• Markers of Collagen Synthesis:
• Procollagen type I N-terminal propeptide (PINP)
 • Procollagen type III N-terminal propeptide (PIIINP)
• Markers of Collagen Degradation:
• Hydroxyproline (in urine and blood)
• Cross-linked telopeptides (e.g., CTX, NTX)
• These markers are used clinically to assess bone turnover, fibrosis, and connective tissue diseases.
Collagenase
• Collagenase is a proteolytic enzyme that specifically cleaves the triple helix of collagen, breaking it down into smaller
peptides14.
• Source: Produced by various cells (e.g., fibroblasts, neutrophils, bacteria).
• Role: Essential for normal tissue remodeling, wound healing, and also in pathological processes (e.g., tumor invasion,
arthritis).
• Regulation: Inhibited by tissue inhibitors of metalloproteinases (TIMPs); retinoic acid inhibits collagenase activity,
preventing excessive collagen breakdown1.
Role of Collagen in Wound Healing
• Early Phase: Collagen is synthesized by fibroblasts to form a provisional matrix that supports cell migration and
angiogenesis.
• Remodeling Phase: Collagen fibers are reorganized and cross-linked to increase tensile strength of the healed tissue.
• Balance of Synthesis and Degradation: Controlled by collagenases and other matrix metalloproteinases to ensure
proper tissue repair without excessive scarring1.
151. Features of the chemical composition of the intercellular matrix of the connective tissue. Glycosaminoglycans,
proteoglycans and glycoproteins. Classification, structure, representatives, Functions.
Ans. The intercellular (extracellular) matrix (ECM) of connective tissue is a complex, dynamic network that provides structural
support, regulates cell behavior, and mediates biochemical signaling. Its main components are fibrous proteins (collagen,
elastin, laminin), glycosaminoglycans (GAGs), proteoglycans, and glycoproteins2.
Glycosaminoglycans (GAGs)
Definition:
GAGs are long, unbranched extracellular heteropolysaccharides, composed of repeating disaccharide units (typically an amino
sugar and a uronic acid)24.
Structure:
• One sugar is an N-acetylated amino sugar (N-acetylglucosamine or N-acetylgalactosamine).
• The other is usually a uronic acid (glucuronic or iduronic acid).
• GAGs are highly negatively charged due to sulfate and carboxyl groups, which attract water and cations, forming a
hydrated gel.
Classification and Representatives:

GAG Structure (Repeating Unit) Main Locations/Functions

Synovial fluid, vitreous humor, ECM;

Hyaluronic Glucuronic acid + N- lubrication, shock absorption, cell
acid acetylglucosamine (not sulfated) migration24
 GAG Structure (Repeating Unit) Main Locations/Functions

Chondroitin Glucuronic acid + N- Cartilage, bone, heart valves; resists

sulfate acetylgalactosamine (sulfated) compression, structural support24

Dermatan Iduronic acid + N-

sulfate acetylgalactosamine (sulfated) Skin, blood vessels, heart valves; flexibility

Keratan Galactose + N-acetylglucosamine Cornea, cartilage, bone; transparency,

sulfate (sulfated) structure

Glucosamine +
iduronic/glucuronic acid (highly Mast cells, liver; anticoagulant, lipoprotein
Heparin sulfated) lipase release24

Glucosamine +
Heparan glucuronic/iduronic acid Basement membranes, cell surfaces;
sulfate (sulfated) filtration, cell signaling2

Functions:
• Provide viscosity, elasticity, and resistance to compression in ECM (e.g., cartilage, synovial fluid)24.
• Regulate cell migration, proliferation, and differentiation (important in development and wound healing).
• Act as barriers to pathogen invasion and as filters (e.g., glomerular basement membrane).
• Heparin acts as an anticoagulant; hyaluronic acid as a lubricant.
Proteoglycans
Definition:
Proteoglycans are large macromolecules consisting of a core protein covalently attached to one or more GAG chains21.
Structure:
• Core protein with multiple GAG side chains (except hyaluronic acid, which is not covalently attached to protein).
• Highly hydrated, forming a gel-like matrix.
• Can aggregate (e.g., aggrecan in cartilage binds hyaluronic acid to form massive complexes).
Classification and Representatives:

Proteoglycan Main GAG(s) Location/Function

Chondroitin sulfate, keratan

Aggrecan sulfate Cartilage; resists compression, hydration2

Dermatan sulfate, chondroitin Connective tissue, regulates collagen

Decorin sulfate fibrillogenesis2

Skin, blood vessels, ECM; cell adhesion,

Versican Chondroitin sulfate migration

Syndecan Heparan sulfate Cell surfaces; cell-matrix interactions, signaling

 Proteoglycan Main GAG(s) Location/Function

Basement membranes; filtration, growth

Perlecan Heparan sulfate factor binding2

Functions:
• Provide compressive strength and resilience (cartilage, intervertebral discs)2.
• Regulate water and ion content of ECM.
• Bind growth factors, modulate cell signaling and adhesion.
• Form selective filtration barriers (e.g., glomerular basement membrane).
Glycoproteins
Definition:
Glycoproteins are proteins with one or more short, branched oligosaccharide chains covalently attached12.
Structure:
• Carbohydrate content is less than in proteoglycans (usually 3–10% of mass).
• Carbohydrates are attached via N-glycosidic (to asparagine) or O-glycosidic (to serine/threonine) linkages1.
Classification and Representatives:

Glycoprotein Location/Function

Fibronectin ECM, plasma; cell adhesion, migration, wound healing2

 Glycoprotein Location/Function

Laminin Basal lamina; cell adhesion, differentiation, filtration2

Integrins Cell membranes; connect ECM to cytoskeleton, signaling

Selectins, lectins Cell membranes; cell-cell recognition, immune response

Functions:
• Mediate cell adhesion to ECM (fibronectin, laminin).
• Guide cell migration and differentiation during development and repair.
• Serve as receptors and signaling molecules (integrins, selectins).
• Regulate immune responses and hemostasis.