Chapter 1

i) Explain about the principles of toxicology.

ii) Discuss the different xenobiotics, routes of entry and their classification.
iii) Explain the different toxic events
iv) Discuss the factors determine the toxicity
v) Explain the dose response curve
vi) Discuss the metabolism and elimination of toxicants

Chapter 2
i) Explain about animal toxicity tests and the rationale behind the use of
animals and testing.
ii) Explain the dose response curve, TD 50, ED 50 and LD 50 doses
iii) Discuss the animal testing OECD test 420, 423 and 425
iv) Discuss the alternative animal testing and different toxicity assays

Chapter 3
i) Explain about lethal synthesis.
ii) Discuss the toxicity of commonly abused drugs
iii) Explain the toxicity of other abused drugs- opioids
iv) Discuss the testing and treatment of drug abuse

 Chapter 1
- Factors that influence toxicity
- Interactions of xenobiotics
 Chapter 2
- Classification system
- Dose response curve
- Alternative tests for animal tests
- Ames’s test
 Chapter 3
- Lethal synthesis
- Paracetamol synthesis
- Screening of drug disorders
- Study about morphine

1
Chapter 1.1
1. Terms:
 Toxicology: the study of the noxious effects of chemical substances
in living organisms.
 Toxic substances: chemicals disturbing biological processes in
detrimental or even fatal ways. [toxicants]
 Dose: amount of compound per unit body weight. [mg/kg].
 Dose effect: Any chemical substance which is present in vast
amounts has the ability to be toxic. Differentiates between therapy
and toxicity.
 Xenobiotics: Excess levels of natural compounds [metabolites,
hormones] which can be harmful. [refers to the action of foreign
compounds]

2. Xenobiotics
 Chemicals not normally produced by the organisms they are foreign
to.
Types of Xenobiotics
 Pollutants: Industrial discharges
 Pesticides
 Pharmaceuticals: drugs, alcohol, tobacco
 Food substances: food toxins, microbials, toxins, allergens,
adverse reactions and additives
 Industrial chemicals: occupational exposure

Major entry routes into body:

 Ingestions: food and drink
 Inhalation: vapors, smokes, inhalants, dusts
 Contact passage of chemicals through the skin
 Other routs: injection, eye drops

Sources of Toxicants
 Contaminated food
 Polluted water

2
  Polluted air
 Pharmaceutical agents
 Drug abuse
 Contact with industrial materials
Ingested Xenobiotics:
 Naturally occurring compounds: flavors, colors
 Food additives: flavors, colors, sweeteners, preservatives
 Polluted in food: dust, microorganisms, pesticide
 Water contaminants

3. Food chains:
 A route termination in food consumed by man.
 Minamata Bay [1952-1960] – 700 cases and 70 deaths due to eating
fish contaminated with methylmercury resulting from industrial
discharge.
 Effects observed: progressive blindness, deafness, loss of
coordination, intellectual deterioration, birth defects in children.

4. Interaction between chemical and organism

 Biological effects of chemical on organism:
 Beneficial: therapeutical, pharmaceuticals
 Toxicological: harmful effects of drug, toxin and pollutants.
 Metabolism of chemical by organism:
 Biotransformation [conversion of chemical to a different form] such
as detoxification [loss of biological activity] and lethal synthesis
[conversion to a toxic form].

5. Types of toxic effect [toxic events]

 Acute Effects
 Develop in seconds, minutes, hours.
 Occur after a single massive dose.

 Chronic Effects
 Result from repeated exposures to small non (sub)- lethal doses.

3
 Exposure can stretch over a long period e.g. Years.

 Local Effects
 Occur at the site of exposure.
 Action ceases when chemical reaction with tissue is complete or
chemical is removed.

 Systemic Effects
 Toxicant is transported & metabolised in the body.
 Terminated by metabolism, binding, storage or excretion.

 Somatic
 Occurs in tissues exposed to toxic agent.
 Directly affects health of the individual exposed.

 Genetic
 Affects the germ cells [Those that generate sperm or eggs].
 Affects the health of descendants.
 Toxicants classed as mutagenic or teratogenic.
 May also result in carcinogenesis and thus adversely affect the health of
individual exposed.
 Effects may be classed as:
1) Reversible where there is no lasting effect.
2) Irreversible where the damage is permanent.

6. Factors that influence toxicity

 Genetic status [e.g. Acetylator phenotype or P450 polymorphisms]
 Body Mass [usually used to calculate dosage]
 Sex [different expression levels of DME, Hormones].
 Age/Degree of maturity
 Health & Well Being [including psychological status and immunological
status].
 Diet

4
7. Factors determining whether a substance is toxic
 Substance Itself.
 Type of Organism exposed to the toxin.
 Amount of Substance (Dose).
 Route of Exposure.
 When > 1 chemical is present
o Actions may be: Simply additive, Synergistic, Potentiative,
Antagonistic.

8. Interactions of Xenobiotics
 Additive - Combined effect = sum of each agent alone: (2 + 3 = 5) e.g.
two organophosphate insecticides.
 Synergistic - Combined effect is > the sum of each agent alone: (2 + 3
=10) e.g. ethanol and CCl4 on liver
 Potentiation - Toxic effect of a compound is greater when a non-toxic
compound is present: (0 + 3 = 10) e.g. Phenobarbital increases the
toxicity of paracetamol.
 Antagonism - Where one compound decreases the toxicity of another
(4 + 6 = 5) e.g. pyrazole decreasing methanol toxicity.

5
Chapter 1.2
1. Toxicokinetic: quantitative relationship linking the various stages of a
toxicant getting to a site of action.
2. What determines the amount of toxicants reaching the
tissues?
 Absorption [route of entry]
 Distribution [movement around body]
 Metabolism [detoxification]
 Excretion [removal from the body]

3. Absorption of toxicants
 Route of entry:
 GI tract: ability to be absorbed, pH factors in GI tract.
 Lungs: size, particularly dusts.
 Skin: ability to pass through skin layers, areas of different skin
thickness.
 Membrane barriers to get to site of action:
 Cell membrane
 Blood-brain barrier
 Placenta
 Modes to cross lipid membrane:
 Diffusion [by hydrophobicity, lipid solubility, ionizability, size, shape,
conc gradient].
 Active transport [by specific proteins, requires the hydrolysis of ATP,
more rapid than diffusion, occur against conc gradient].

4. Distribution of toxicant
 Once in bloodstream, toxicant must reach site of action.
 Influenced by the chemical itself, the mode of transport.
 Conc at the site of action depends on plasma concentration.

6
5. Factors reducing the level of free toxicant
 Extracellular proteins:
 [Albumin] – transports endogenous substances like bilirubin and fatty
acids, has broad affinity for many chemicals, sequesters toxicants
[reduced free conc].
 Intracellular proteins
 [Hemoglobin] – abundant red cell protein, has many groups which can
react with toxicants.
 [Ligandin] – found in liver, kidney tubules, intestinal mucosa, binds
lipophilic and ionic substances, a glutathione transferase, binds and cell
transport of many lipophilic substances and removal of electrophiles by
catalyzing their reaction with nucleophilic glutathione.
 [Metallothionein] – found in liver and kidney, has structure suited for
binding metals, normally binds zinc and copper, also calcium and
methyl mercury.
 Storage in adipose tissue
 Metabolism
 Blockage of one or more pathways from: inhibition of enzymes,
damage to structural organization.
 Causes of depletion of ATP:
 The action of uncouplers/ inhibitors of oxidative phosphorylation.
 Hypoxia.
 Inhibition of electron transport.
 Decreased NADH.
 Physical damage to the mitochondria.
 Inhibition of ATP supply [mechanisms]
 Reduced oxygen delivery to tissue [CO2, Nitrite]
 Inhibition of carbohydrate metabolism [Fluoroacetate]
 Inhibition of electron transport [rotenone, CO]
 Uncoupling of oxidative phosphorylation [Nitrophenol]

7
  Inhibition of ATP formation [Oligomycin]
 Structural damage to mitochondrion

 Consequences of lack of ATP

 Decrease in active transport [sodium, potassium, and calcium
pumps, reabsorption of essential amino acids and glucose in kidney
tubules]
 Decreased synthesis of proteins, carbohydrates, lipids, nucleic acids.
 Inability to maintain and regulate cellular processes [membrane
integrity of RBC = Hemolysis].
 Excretion of toxicant
 Polar compounds are easily removed, aided by metabolism
[detoxification and lethal synthesis].

8
Chapter 1.3
1. Detoxification
 Harmful chemical [ingested, absorbed, inhaled] are converted to less
harmful chemicals [more easily excreted] by using enzymes [in
endoplasmic reticulum, ‘microsomal enzymes’, cytochrome P450, in
cytosol].
 Transformation of lipophilic molecules into more acidic and water-
soluble molecule.

 Xenobiotic metabolism [characteristics]: increases solubility in

water, more acidic, shorter half-life, shorter exposure, no accumulation,
structural change.

 Metabolism
 Phase 1: Biotransformation [additional of functional groups: oxidation,
reduction, hydrolysis, hydration, dehalogenation  metabolites
excreted]
 Phase 2: Conjugations [synthetic reactions – glutathione, glucuronic
acid, sulphate, amino acid  make a more polar substance,
transferases is the enzymes facilitated in this phase]
 Phase 3: Bacterial metabolism in gut, transport out of cell, absorption
into urine.

 Phase 1 – Oxidations
 Carried out by oxidases such as mono-oxygenases.
 Found in animals, plants, microbes, present in endoplasmic reticulum 
in liver cells.

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 Mono-oxygenases  enzymes insert one of the two oxygen atoms from
one molecule of O2 into substrate. The other oxygen atom is reduced to
water.
 Cytochrome P450 catalytic cycle: broad specificity, lipophilic substrate,
more active when bound to SER, tissue specificity, and cyclical reaction.
o Reactions: hydroxylation, dealkylations, epoxide formation,
substitution of P=S by P=O, formation of sulphoxides and
sulphones, and oxidative deamination.

 Non-microsomal oxidation:
 Amine oxidation [release amine groups]
 Alcohol oxidation [alcohol to aldehyde]
 Aldehyde oxidation [aldehyde to organic acid]
 Purine oxidation
 Phase 1 – Reduction: azo reduction, nitro reduction
 Phase 1 – Hydrolyses: esterases, amidases
 Phase 1 – Dehalogenations: reductive dichlorination,
dehydrochlorination
 Phase 1 – END PRODUCTS: more polar than substrate. [further
metabolism by conjugation or if it hydrophilic  excreted directly]

 Phase 2 – Conjugation reactions

 Less lipophilic [more polar], less toxic, easier to excrete.
 Glucuronide formation
o Activation of glucuronic acid
o Conjugation to reactive group
 Sulphation
o Addition of sulphate groups to -OH groups
o Catalyzed by sulphotransferases in cytosol
o Products are very water soluble, nontoxic and easily excreted.
 Aminoacidic conjugation
o Addition of aminoacidic to -COOH.
o More stable than sulphate and glucuronide in environment

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 Glutathione conjugation
o Hydrolyzed to Cys conjugate
o Acetylated to a mercapturic acid
o Does not require activation
o Nucleophilic sulphydryl [-SH] group neutralizes toxic electrophiles

Chapter 2.1
1. Dose response curve

• Lethal Dose for 50% Mortality (LD50)

– Statistically derived single dose  death in 50% of a given population
of organisms.
– Not an exact value.
– May vary from species to species.
– Only use in a comparative sense i.e. compare toxicity of one
substance vs another

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  Therapeutic Index of drugs: LD50/ED50
TD50/ED50
 Require this value to be as high as possible.

2. Alternatives to the LD50 test

 New OECD Guidelines:
 OECD: Organisation for Economic Co-operation and Development:
Environmental Health & Safety Programme  responsible for
reviewing guidelines for chemical toxicity test considering new
scientific data.
 420 which does not require death as endpoint
 423 which requires deaths <3
 425 which requires deaths <3

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  Key to EU classification:
T* = very toxic
T = toxic
H = harmful
U = unclassified

 OECD Test 420

 Normally start with a sighting study
o Uses a single animal per dose
o Starting dose based on information from literature
o Use same doses as Test 420.

 Key outcomes
 A =  2 deaths
 B =  1 with toxicity or no death
 C- = no toxicity

13
  OECD Test 423
 Same doses as OECD 420 test
o Uses 3 animals/step [not 5]

 Key outcomes
 No further testing required
 Same dose 3 more animals
 Next higher or lower dose to 3 additional animals

 OECD Test 425

 Uses a maximum of 5 animals
 A sequential dosing method
 First animal given dose a step below LD 50
 Monitor for 48 hrs before next step

 Key outcomes
 Survives – next animal increase dose by factor of 3.2
 Dies – next animal reduce dose by similar factor
 Progression is by a series of up and down step

14
Chapter 2.2
1. Alternatives to animal tests
 In Vitro
o Isolated enzymes and organelles
o Micro-organisms
o Tissue culture cells
o Isolated organs
o Artificial systems (organs-on-chips, 3-d human cell-derived
model)
 Lower Forms of Animal and Plant Life
 Quantitative structure-activity relationship (QSAR) and other
Computational Methods (e.g. in silico)

15
  Human-patient simulators*

2. Mutagenicity assay
 In vitro tests:
o Bacterial assay for mutation e.g. Ames Test, SOS Chromo Test
o Chromosomal aberrations and clastogenicity
o Gene mutations in mammalian cells
 In vivo tests:
o Bone marrow micronucleus test
o Tests for DNA damage in various tissues e.g. COMET test
 Germ Cell Assays:
o Interaction with DNA
o Potential for inherited effects
o Assessment of heritable effects

Why test for mutagenicity

 Genetic changes are central to the development of cancer
o Genes, chromosomes, chromosomal structure
 Induction of mutation in germ cells
o Congenital abnormalizes
o Inheritable diseases
o Spontaneous abortions
 Most mutagens shown to be carcinogenic
 Chemical mutagens – test for genetic alterations in bacterial and
mammalian cells.
3. Bacterial assay for mutation
• Large population can be culture and treated
• Good understanding of bacterial genetics
• Assays are quick and cheap
• Methods are well validated
• Strains have been developed with enhanced sensitivity

AMES test [reverse mutation test]

16
Auxotrophs are a group of organisms that lost the ability to synthesize
certain substances required for their growth owing to the presence of
mutations.

 Cell wall mutation  most chemicals to enter the cell.

 Mutation (uvrB) eliminating ability to repair DNA damage.
 + plasmid (pKM101) which stimulates the conversion of DNA
damage into inheritable mutations.
 Indirect method as observing phenotype change.

SOS chromotest
 Several substrates with can be used in quantitative assays
o X Gal which gives blue color on hydrolysis = Abs 609nm
o o-nitrophenyl-b-galactopyranoside gives yellow color on
hydrolysis = Abs 405-410nm
o Methylumbelliferyl-b-D galactoside, fluorescence measured at
340/460 nm

17
4. -Galactosidase assays
 Found in many organisms including bacteria, yeast, and mammals.
 Involved in the breakdown of lactose and other galactosidase into
glucose and galactose.
 Used in applications like gene expression analysis, microbial
identification, cellular metabolism etc.
 To measure the levels of active β-galactosidase expressed in cells
transfected with plasmids expressing the lacZ gene (a reporter
construct).

 FluoroMetPlate Assay - A sensitive fluorometric assay was developed

for the enumeration of cells in microtiter plates. This assay is based on
the fluorescence enhancement of propidium iodide (PI) upon
binding with double-stranded nucleic acids.
 The lac Z gene is a gene that encodes for β-galactosidase enzyme in
the bacterium Escherichia coli (E. coli). The lac Z gene is part of the lac
operon, a cluster of genes involved in the metabolism of lactose.

5. Tests for mutagens by chromosome damage

• Chromosome Aberrations:
• Used to identify agents that cause structural changes in
chromosomes.
• Cultures of mammalian cells e.g. CHO (Chinese Hamster Ovary
Cells), fibroblasts, peripheral blood lymphocytes.

18
 • Exposed to test substances with or without biotransformation
(S9 fraction).
• Expose then arrest in metaphase with agent such as Colchicine
or Colcemid.
• Stain chromosomes and scan for deletions, fragments etc.
• Sister Chromatid Exchange:
• Cultured mammalian cells e.g. Human leucocytes, CHOs.
• Add test chemical and allow 2 rounds of replication in presence
of 5-bromodeoxyuridine (BrdU).
• Stop at 2nd metaphase with colchicine.
• Stain with Giemsa.
• Differential staining of chromatids.
• In vivo test similar, analysis of bone marrow cells.
• Mutagens greatly increase the number of SCEs within cells.
• Good correlation between frequency of SCE & mutagenicity.
• Micronucleus Test
• Micronuclei – small nuclei which are separate from & additional
to main nucleus.
• Produced during telophase of mitosis (meiosis).
• Correspond to daughter nuclei which are broken or lagging in
formation thus not included in main nucleus.
• Test - Dose mice twice in 24hr period, collect samples 30hr
later.
• Collect bone marrow peripheral blood
• Analyse erythrocytes (RBC)
• Erythroblast to erythrocytes = extrusion of the nucleus
• Micronucleus will remain in erythrocyte.
• Score 1000 cells per animal

• COMET Assay
• Single cell gel electrophoresis assay to detect DNA strand
breaks.
• Test involves expose of animals or cells to chemical.

19
 • Cell suspended on agarose on slide.
• Cell lysed  DNA exposed.
• Soak in neutral or alkaline buffer to loosen DNA mass, allows
broken DNA to unwind.
• Electrophoresis.
• Damaged DNA strand is pulled in one direction  ‘COMET’.

• Gene Mutation Test in Mammalian Cells

• Mammalian TK Test – similar idea to Ames Test.
• Use mouse lymphoma cells which are heterogeneous for TK
locus i.e. TK+/-.
• Detects point mutations, deletions, translocations, mitotic
recombination and aneuploidy.
• Cells deficient in TK are resistant to cytostatic effects of TFT
(trifluorothymidine), a pyrimidine analogue.
• Mutants cells proliferate in the presence of TFT.
• Mutational frequency derived from number of colonies in
selective medium.

6. Replacements for eye and skin sensitivity tests

 Draize test
o Alternative test: Corrositex assay
o Dye uptake of cells:
 MTT assay – yellow dye converted to blue formazan by
mitochondrial dehydrogenases thus ↓ blue ↓ cell viability.
 Trypan Blue – vital stain which is excluded from living cells i.e.
dead cells take up dye.
 Fluorogenic fluorescein diacetate – cleaved by esterases
produced in living cells to produce fluorescence compound.
 Fluorescein permeability – enters cells due to damage of cell
barrier, measure internal fluorescence.

20
  Neutral red dye uptake – taken up by lysosomes ↓ dye uptake
↓ cell viability.
 Lactate dehydrogenase leakage – increased in medium due to
damage of membrane.
 Artificial skin

Chapter 3
1. Lethal synthesis
 Inappropriate metabolism of a relatively safe xenobiotic to a more
toxic molecule.
21
  Origin is broad specificity of enzymes.
 Many enzymes show tissue specificity – localized toxin production.
 All tissue contains xenobiotic metabolizing enzymes, but most
important organ is the liver.
 Hepatotoxicity: only site of metabolism and exposure to highest
concentrations.

2. Paracetamol synthesis [non-narcotic analgesics]  acts chiefly on

peripherally.
- Widely available analgesic.
- Safe when used at therapeutic dose.
- Overdose ® hepatotoxicity and liver necrosis.
- Major route of metabolism is by conjugation.
- Remove about 60-90% of drug.
- 5% free per each pass through the liver.
- Toxicity arises from an alternative route of metabolism.
- Facilitated by P450s.
- In overdose, glutathione becomes depleted.
- P450 rxn very efficient – produces NAPQI.
- Binds to liver protein – necrosis.
- Treatment
 Inhibit P-450 using Cimetidine.
 Increase glutathione synthesis.
 Provide sulphate for conjugation using methionine.
 Provide glutathione alternative with n-acetyl cysteine.

3. Morphine [narcotic analgesics]  acts chiefly in the CNS.

 Main effects of morphine on the CNS:

22
 o Depression, leading to analgesia, respiratory depression
(decrease in sensitivity of the respiratory center to PCO 2),
depression of cough reflex, sleep).
o Excitation, leading to vomiting, miosis (pupil constriction),
convulsions (very rare).
o Changes of mood – euphoria (sense of well-being) or
dysphoria.
o Tolerance and dependence (psychological and physical)
o Smooth muscle stimulation: Gastrointestinal muscle spasm
(with constipation) and biliary tract spasm.
o Cardiovascular system: Dilation of resistance and
capacitance vessels.
o Other effects: Sweating, histamine release, pruritus,
piloerection, antidiuretic effect.

Psychosocial manifestations of mild/moderate drug disorders

 Psychological/Behavioral
o Agitation, irritability, dysphoria, difficulty in coping, mood
swings, hostility, violence, psychosomatic symptoms,
hyperventilation, generalized anxiety, panic attacks,
depression, psychosis.
 Family
o Chronic stable family dysfunction, marital problems, anxiety
and depression in family members, divorce, abuse and
violence.

23
  Social
o Alienation and loss of old friends, gravitation toward others
with similar lifestyle.
 Legal
o Arrests for disturbing the peace or driving while intoxicated,
stealing, drug dealing.
 Financial
o Borrowing or owing money, selling personal or family
possessions.

4. Screening and assessment

 Every person over 60 should be screened for alcohol and drug abuse
as part of regular physical examination. “Brown Bag Approach”
 Screen or re-screen if certain physical symptoms are present or if the
older person is undergoing major life transitions.
 Ask direct questions about concerns.
o Preface question with link to medical conditions of health
concerns
o Do not use stigmatizing terms (i.e. drug addict)

 Special assessment
 Functional Abilities
o Activities of Daily Living (ADLs)
o Instrumental Activities of Daily Living (IADLs)
o SF-36
 Comorbidities
o Physical
o Psychiatric
 Affective disorders
 Suicide risk
 Sleep Disorders
 Cognitive Impairments
o Dementia

24
  Orientation/Memory/Concentration Test
 Folstein Mini-Mental Status Exam (MMSE)
o Delirium
 Confusion Assessment Method (CAM)
o Other cognitive impairments
 Trauma from falls, MVA, accidents
 Wernicke-Korsakoff syndrome

1. Briefly elaborate paracetamol toxicology and its treatment.

2. Discuss TWO (2) opioid analogs and potential treatment.
 Fentanyl and Etorphine, are two potent opioid analogs associated
with a high risk of overdose and respiratory depression.
 Fentanyl, approximately more than 80 times more potent than
morphine, is commonly used for anesthesia [severe pain
management] but poses a significant overdose risk.
 Treatment of fentanyl overdose primarily involves naloxone
administration to reverse opioid toxicity, along with supportive care
for respiratory and cardiovascular complications.
 Etorphine, even more potent than fentanyl at approximately more
than 1000-3000 times the potency of morphine, is primarily used
with acepromazine as a large animal tranquilizer leading to
numerous overdose deaths.
 Treating etorphine overdose requires aggressive
naloxone/naltrexone administration and intensive supportive
care due to its extreme potency and rapid onset of respiratory
depression.

25
  Major concerns of using opioid treatments:
 Addiction
 Tolerance
 Neuropsychological effects:
o Psychomotor performance, cognitive function, affective
disturbance.
o Opioids lower testosterone levels in animals, heroin addicts,
methadone maintenance pts, and intrathecal opioid pts.
o Opioids anecdotally produce loss of libido and impotence in
men; amenorrhea and infertility in women.
o Low testosterone: fatigue, loss of muscle mass, mood
disturbances, osteoporosis

 Symptoms
– Nausea, vomiting, constipation
– Dizziness, sweating
– Itching, etc.

Criteria for substance use disorders

• Using more of a substance than intended or using it for longer than
you’re meant to.
• Trying to cut down or stop using the substance but being unable to.
• Experiencing intense cravings or urges to use the substance.
• Needing more of the substance to get the desired effect — also
called tolerance.
• Developing withdrawal symptoms when not using the substance.
• Spending more time getting and using drugs and recovering from
substance use.
• Neglecting responsibilities at home, work or school because of
substance use.
• Continuing to use even when it causes relationship problems.

26
 • Giving up important or desirable social and recreational activities
due to substance use.
• Using substances in risky settings that put you in danger.
• Continuing to use despite the substance causing problems to your
physical and mental health.

 Chapter 4
- Explain about lethal synthesis
- Discuss the toxicity of ethanol – drug of abuse
- Explain the testing of ethanol
- Discuss the toxicity of methanol, EG and isopropanol
 Chapter 5
- Explain about the sources of CO
- Discuss the properties and effects of CO
- Explain the action of CO toxicity and system effects
- Discuss the testing of CO poisoning and management of CO poisoning
 Chapter 6
- Discuss the toxic agents in the environment
- Discuss bioaccumulation and biomagnification
- Explain the action of few environmental toxicants’ epidemiology
and animal testing
- Discuss the factors affecting environmental toxicity
 Chapter 7
- Explain about the toxicants in food
- Discuss the different natural toxins from food sources
- Explain the different natural contaminants of food sources
- Discuss with names examples
 Chapter 8
- Discuss the role of toxic chemicals, rodenticide, heavy metals in
toxicity
- Describe the mechanism driven by the metals for causing
toxicity in major system
- Explain the toxic chemicals, chemical warfare agents and
their effects
 Chapter 9

27
 - Explain about genotoxicity and teratogenicity with at least one
example
- Discuss the classes and effects of genotoxic chemicals and
thalidomide
- Explain the mechanism of thalidomide causing teratogenicity
- Discuss the treatment and prevention of teratogenicity in general

Chapter 4
1. Explain about lethal synthesis

- Broad specificity of enzymes originates from their ability to act on

various substrates.

- Many enzymes exhibit tissue specificity, leading to localized toxin

production.

- Xenobiotic metabolizing enzymes are present in all tissues, but the

liver is the most crucial organ.

- Hepatotoxicity occurs because the liver is the primary site of

metabolism and is exposed to the highest concentrations of toxins.

- Role of the liver

- Products of digestion are transported to the liver via the portal

circulation through the hepatic portal vein.

- The liver acts as a sentry towards xenobiotics.

- Components required by the body are allowed to pass through.

- Harmful compounds are filtered out through 'First pass' elimination.

- Hepatotoxicity may occur due to 'lethal synthesis' in the liver.

28
2. Discuss the toxicity of ethanol – drug of abuse
- Ethanol is the most commonly abused substance due to its ability to
relax individuals, increase sociability, and reduce inhibitions.
- While ethanol can produce pleasurable effects, it also causes significant
physical damage.
- Ethanol is considered a xenobiotic, which undergoes metabolic
detoxification in the body.
- Ethanol toxicity can manifest both acutely and chronically, with
different physiological and clinical effects.

Acute Ethanol Toxicity

- Acute ethanol toxicity is observed as an increased response to GABA, a
- The effect of ethanol on the CNS depends on the blood alcohol
concentration (BAC), as described by the Miles scale:
o 30 mg/100 mL: Mild euphoria.
o 50 mg/100 mL: Mild uncoordination.
o 100 mg/100 mL: Difficulty in walking.
o 200 mg/100 mL: Confusion and drowsiness.
o 300 mg/100 mL: Stupor.
o 400 mg/100 mL: Deep anesthesia.
o 500 mg/100 mL: Death.
- Clinical Effects of Ethanol
- Ethanol can be detected in urine up to 12 hours after consumption,
and its consumption can lead to addiction.
- Clinical Symptoms:

29
 o Intoxication.
o Loss of motor control.
o Lethargy (fatigue).
o Aggressive behavior.
o Stupor.
o Alcohol odor on breath and from sweat glands.
o Dulling of nerve endings, slowing body functions, and impairing
performance.
- Ethanol Metabolism
- Ethanol metabolism leads to the production of ketone bodies rather
than glucose.
- The liver metabolizes ethanol at a rate of 100 mg/kg/hr., with a small
amount metabolized by the stomach wall and mucosal lining.
- Metabolic Pathway:
o Ethanol (CH3CH2OH) is converted to acetaldehyde (CH3CHO) by
alcohol dehydrogenase (in the cytosol).
o Acetaldehyde is further metabolized to acetic acid (CH 3COOH)
by aldehyde dehydrogenase (in the mitochondria).
o Acetic acid is then further metabolized by acetyl-CoA
synthetase.
o Cytosolic Isoform in the liver: Acetyl-CoA is used for cholesterol
and fatty acid synthesis.
o Mitochondrial Isoform in other cells: Acetyl-CoA enters the TCA
cycle.
- Excess alcohol consumption increases the NADH/NAD+ ratio,
leading to metabolic disturbances such as:
o Lactic Acidemia.
o Hepatic Steatosis (fatty liver).
o Ketoacidosis.
o Hypoglycemia.
o Dehydration due to suppression of ADH.

Chronic Ethanol Toxicity

30
  Chronic ethanol consumption leads to the induction of xenobiotic
metabolizing enzymes [tolerance], increasing the metabolism of
other drugs.
 Individuals may develop dependence, experiencing hangovers and
preventing withdrawal symptoms.
 Chronic ethanol abuse can result in loss of appetite and vitamin
deficiencies, particularly thiamine deficiency.
 A severe form of alcohol withdrawal that can include confusion,
tremors, and hallucinations.
 Chronic ethanol toxicity leads to liver damage, manifesting as
hepatitis, hepatomegaly, fatty liver, and cirrhosis.

3. Explain the testing of ethanol

- Breath Testing: Uses infrared spectrometry to measure the amount of

alcohol on the breath, from which blood alcohol levels are inferred.
- This method provides immediate results and is advantageous for
showing current use.
- The results can be correlated with blood level (if alcohol level is 0.040
or greater, will require a completion of a Substance Abuse Professional
Evaluation).
- However, it requires specialized equipment which are expensive,
requires maintenance and quality control.

- Blood Testing: Considered the most accurate method but is invasive.

- Blood Alcohol Concentration (BAC) is expressed as a percentage, with
specific thresholds indicating levels of impairment.
- For instance, a BAC of 0.08% is the level of presumptive driver
impairment in many states.
- Urine Testing: While urine tests have an established collection
routine, their correlation to blood levels is less accurate.
- The alcohol level in urine can be 1.3 times that in blood after peaking,
typically two hours post-drinking. T

31
- here is also a risk of in situ fermentation in urine samples, which can
lead to falsely high alcohol levels.
- Saliva Testing: Provides immediate results and represents newer
technology.
- While it is useful, especially for alcohol testing, it is less commonly used
than breath or blood tests.

4. Discuss the toxicity of methanol, EG and isopropanol

- Methanol: A cheaper alternative to ethanol, methanol's toxicity arises
from its conversion to formic acid.

- Ingestion of as little as 10 mL can cause blindness, and 30 mL can be

fatal [death].
- Methanol poisoning symptoms progress from nausea, headache, dizzy,
and abdominal pain [8-36 hrs.] to unconsciousness, coma, and death
[>36 hrs.]

- Ethylene Glycol (EG): Commonly found in antifreeze.

- Ethylene glycol is sweet-smelling and can be toxic when ingested.
- Its toxicity progresses through three stages.
o Stage 1: 30 mins to 12 hrs.  intoxication and nausea.
o Stage 2: 12 hrs. to 24 hrs.  tachycardia, pulmonary oedema

32
 o Stage 3: 1 to 3 days.  renal necrosis, acute renal failure, death
- Ethylene glycol primarily metabolized in liver by the enzyme alcohol
dehydrogenase to glycolaldehyde.
- Glycolaldehyde is further metabolized by aldehyde dehydrogenase to
glycolic acid.
- Glycolic acid is converted to glycolic acid by the action of various
enzymes including lactate dehydrogenase.
- Glyoxylic acid undergoes oxidation to form oxalic acid.
- Consequences:
- Oxalic acid binds with calcium in the blood to form insoluble calcium
oxalate crystals. The formation of these crystals leads to hypocalcemia.
- Calcium oxalate crystals can deposit in the renal tubules, causing
physical blockage, inflammation, and necrosis of renal tissues. This can
lead to acute kidney injury and renal failure.
- The metabolism of ethylene glycol produces toxic metabolites that
inhibit normal metabolic processes  buildup of organic acids.
- Calcium oxalate and aldehyde metabolites can reduce cerebral
function, leading to symptoms such as confusion, seizures, and coma.

- Isopropanol: Typically used as a disinfectant, isopropanol is more toxic

than ethanol but is often abused similarly.
- Its effects include central nervous system depression, and ingestion can
lead to symptoms such as nausea, vomiting, abdominal pain, and in
severe cases, respiratory depression and coma.

Chapter 5
1. Explain about the sources of CO.
- Motor Vehicle Exhaust: Running engines in closed spaces or with
faulty exhaust systems can lead to CO accumulation.

33
- Propane-Powered Equipment: Devices such as lifts, water heaters,
and concrete saws can produce CO.
- Combustion for Heating or Cooking: Using camping equipment or
heating systems in enclosed spaces can generate CO.
- Smoke Inhalation in Fires: CO is a significant component of smoke
from fires.
- Methylene Chloride: This industrial product, used as a paint and
adhesive remover, is slowly metabolized to CO in the liver, leading to
CO poisoning upon prolonged exposure.

2. Discuss the properties and effects of CO.

Properties:
- CO is an odorless, colorless, tasteless, and non-irritating gas, making it
undetectable without specialized equipment.
- It is a flammable gas with an auto-ignition temperature of 1128 °F and
explosive limits ranging from 12.5% to 74%.
- A natural by-product of incomplete combustion from equipment burning
carbon-based fossil fuels such as gasoline.
- CO has a vapor density of 0.968, which is slightly less than air, causing
it to rise with warm air and disperse evenly once it cools.
- The half-life of carbon monoxide (CO) is 4 to 6 hours in 21% oxygen, 80
minutes in 100% oxygen, and 22 minutes with hyperbaric oxygen
treatment.

Factors that determine the toxicity of CO

- Initial HbCO concentration
- Concentration of CO inhaled
- Length of exposure
- Activity while inhaling CO
- Body size and physiological factors
Effects:
- General symptoms: headache, nausea, vomiting
- Cardiovascular effects: chest pain, tachypnea, tachycardia,
hypotension, arrhythmia, cardiac arrest.

34
- Neurological effect: dizziness, ataxia, seizures, coma.
- Other effects: metabolic acidosis, retinal hemorrhages.
- Delayed neuropsychiatric syndrome occurs in 10-30% victims, 3—
240 days post exposure. Dementia, parkinsonism, personality changes
and cognitive changes.

Pathophysiology:
- CO is colorless, odorless, non-irritant toxic gas. CO toxicity due to
cellular hypoxia and direct cellular injury.
- Cellular Hypoxia:
- CO competes with oxygen for binding to hemoglobin with an affinity
200-250 times greater than oxygen, leading to impaired oxygen release
and cellular hypoxia.
- Direct Cellular Injury:
- Includes reoxygenation injury in the CNS, lipid peroxidation, and
free radical formation.

- CO toxicity in pregnancy:
- Fetal hemoglobin binds CO more readily than adult hemoglobin,
increasing the risk of fetal injury.

- Cellular level pathophysiology

- Extravascular Heme-Containing Proteins: About 15% of CO binds
to extravascular heme proteins.
- Cytochrome Oxidase (cytochrome aa3): CO binding affects ATP
production, causing intracellular acidosis, which can persist after
exposure.
- Cardiac and Skeletal Myoglobin: CO binding at COHb levels as low
as 2% alters tissue oxygen uptake.
- Tissue Hypoxia:
35
- Formation of Carboxyhemoglobin (COHb): CO binds with
hemoglobin to form COHb, reducing oxygen transport.
- High Affinity: Hemoglobin's affinity for CO is 250 times greater than
for oxygen, impairing oxygen release at the tissue level.
- Increased Ventilation: The body increases ventilation, leading to
further CO uptake.

- Cardiovascular Pathophysiology:
- Myocardial Depression: Caused by hypoxic stress, cytochrome aa3
dysfunction, and CO binding to cardiac myoglobin.
- Arterial Hypotension: Resulting from myocardial depression and
nitric oxide (NO)-related peripheral vasodilation.
- Loss of Consciousness (LOC): Due to reduced cerebral perfusion and
ischemic reperfusion injury.

- Neurovascular Pathophysiology:
- Nitric Oxide (NO) Increase: CO in circulation causes a significant
increase in NO in perivascular tissues, released by vascular endothelial
cells and platelets.
- Oxygen Radicals: Impaired mitochondrial function leads to oxygen
radical production, which reacts with NO to form peroxynitrite (ONOO-).
- Peroxynitrite Damage: This compound binds to perivascular tissue
proteins, causing injury and increased capillary permeability in the CNS
and pulmonary vascular beds.
- Endothelial Injury: Leads to the expression of adherence molecules
(beta 2 integrins), allowing leukocytes to bind to the injured
endothelium, reducing cerebral perfusion, and initiating CNS lipid
peroxidation.

- CO Poisoning in Pregnancy:
- Neurologic Abnormalities and Stillbirth: High incidence due to
CO’s strong binding to fetal hemoglobin, which absorbs and eliminates
CO more slowly.

36
- Hyperbaric Oxygen (HBO) Therapy: Considered safe during
pregnancy, though no established guidelines exist for its use, except
when COHb levels exceed 15%.

3. Explain the action of CO toxicity and system effects

4. Discuss the testing of CO poisoning and management of CO
poisoning Testing:

- High Level of Clinical Suspicion:

- Diagnosis often begins with a high index of suspicion, especially when
patients present with nonspecific symptoms like headache, dizziness,
nausea, or altered mental status, particularly if there's a history of
potential CO exposure.

- Measurement of serum COHb Levels:

- The measurement of carboxyhemoglobin (COHb) in the blood is critical
for confirming CO poisoning. Elevated COHb levels indicate the extent
of CO exposure.
- COHb can also be measured in exhaled breath, providing a non-
invasive method for initial assessment.
- COHb Levels:
o Normal ranges are <3% for non-smokers and <10% for
smokers.
o COHb levels do not reliably predict the severity of poisoning or
correlate well with symptoms, making them a poor indicator of
clinical outcome.
- Arterial Blood Gas (ABG): Metabolic acidosis, often due to lactate
accumulation, can be detected.
- Electrocardiogram (ECG): Used to identify ischemia and arrhythmias,
which may occur as a result of CO poisoning.

- Spectrophotometry:
- COHb levels are typically measured using spectrophotometry, a
technique that detects the specific wavelengths of light absorbed by
COHb in the blood.

37
- Limitations of Pulse Oximetry:
- Principle: Pulse oximetry measures oxygen saturation (SaO2) by
detecting light absorption at two wavelengths (660 nm for
deoxygenated hemoglobin and 940 nm for oxygenated hemoglobin).
- Limitations: It is unreliable in the presence of significant levels of
abnormal hemoglobin such as methemoglobin (MetHb),
carboxyhemoglobin (COHb).
- Pulse oximetry tends to overestimate true arterial oxygen saturation
when COHb is present because it cannot distinguish between
oxyhemoglobin (HbO2) and COHb.
- Elevated COHb levels can falsely elevate the reported SaO2, leading to
a misinterpretation of the patient’s oxygenation status.

- Comprehensive Neurological and Neuropsychological

Assessment:
- A thorough neurological examination is essential to assess for any
immediate or delayed effects of CO poisoning, including cognitive and
motor deficits.
Psychometric Testing:
- Challenges: Psychometric testing in CO poisoning is subjective and
may be influenced by factors such as the practice effect (improvement
due to repeated testing) and the gradual resolution of other toxic
effects over time.
- Persistent or delayed neurologic sequelae are more likely in patients
with abnormal psychometric test results.
- Abnormal psychometric testing could potentially indicate the need for
hyperbaric oxygen (HBO) therapy, even in cases of mild CO poisoning,
though this is still debated.

- The CO Neuropsychological Screening Battery may be employed

to evaluate cognitive function and identify subtle neuropsychological
impairments that could result from CO exposure.

38
- Imaging Studies:
- A CT scan of the brain may be conducted to exclude other conditions
that could mimic the symptoms of CO poisoning, such as stroke, brain
hemorrhage, or other structural brain abnormalities.

Management:
- 100% Oxygen ASAP: Administer immediately to reduce COHb
levels.
- Arterial Blood Gas (ABG) Analysis: Check for metabolic acidosis.
- Carboxyhemoglobin (COHb) Levels: Monitor to assess treatment
effectiveness.
- Electrocardiogram (ECG): Evaluate for ischemia and arrhythmias.
- Chest X-Ray (CXR): Rule out respiratory complications (e.g.,
pulmonary edema).
- Cardiac Enzymes: Detect myocardial injury.
- Cardiac Monitoring: Continuous monitoring for arrhythmias.
Treatment of Carbon Monoxide (CO) Poisoning
- High-Flow Oxygen:
- FiO2 ~100%: Administer norm baric oxygen to reduce COHb levels.
- Half-Life Reduction:
 21% O2: 4-6 hours
 100% O2: 40-80 minutes
 100% O2 at 2.5 atm (Hyperbaric): 15-30 minutes
- Oxygen Therapy Continuation: Continue until COHb levels
normalize.
- Consider Concomitant Injuries: Be aware of potential smoke
inhalation and burn injuries.
- Normobaric vs. Hyperbaric Oxygen Therapy:
- Hyperbaric Oxygen (HBO): Accelerates resolution of acute
symptoms.
- Evidence for HBO: Unclear effect on late complications and mortality.
- Hyperbaric Oxygen Therapy Benefits
 Speeds up CO removal from the bloodstream.

39
  Increases oxygen delivery to tissues.
 Facilitates the dissociation of CO from cytochrome oxidase.
 adhesion of leukocytes to vascular endothelium.
 Reduces CNS lipid peroxidation.

Problems in CO Poisoning
 Lack of reliable methods to predict the severity of CO poisoning.
 Difficulty comparing study results due to lack of standardized
disease staging.
 Presence of myths and inaccurate information in the literature.
 Often neglecting timely diagnosis and prevention of further CO
exposure.

Prevention of CO Poisoning
 Raise awareness about CO poisoning risks and prevention.
 Identify and address activities at risk for CO exposure.
 Educate workers on the safe use of propane-powered tools.
 Ensure proper ventilation in confined spaces to prevent CO buildup.
 Install CO detectors in industrial and domestic settings.
 Report cases to public health services for proper monitoring and
response.
Chapter 6
1. Discuss the toxic agents in the environment
- Synthetic and Natural Toxicants: One type of environmental health
threat.
- Other Environmental Health Threats:
- Physical Hazards: Floods, blizzards, landslides, radon, UV exposure.
- Chemical Hazards: Disinfectants, pesticides.
- Biological Hazards: Viruses, bacterial infections.
- Cultural or Lifestyle Hazards: Drinking, smoking, poor diet, crime in
the neighborhood.

- Types of environmental toxicants

- Carcinogens: Cause cancer.

40
- Mutagens: Cause mutations in DNA.
- Teratogens: Cause birth defects.
- Allergens: Trigger unnecessary immune responses.
- Neurotoxins: Damage the nervous system.
- Endocrine Disruptors: Interfere with hormonal functions.

2. Discuss bioaccumulation and biomagnification

- Biomagnification and Bioaccumulation
- Biomagnification refers to the increasing concentration of toxic
chemicals at each trophic level in a food chain. As predators
consume prey, they accumulate all the toxicants the prey has ingested,
leading to higher concentrations in top predators.
- Bioaccumulation occurs when an organism absorbs a toxic substance
faster than it can excrete it, leading to a buildup of the substance in its
tissues over time.

- Persistent Organic Pollutants (POPs)

- Characteristics of POPs:
- POPs break down very slowly due to their stable chemical structure,
which allows them to remain in the environment for long periods.
- These chemicals dissolve in fats [fat soluble] rather than water,
leading to their accumulation in the fatty tissues of organisms.

- Why POPs Accumulate in Cold Climates:

- In colder regions, microbial activity that would typically break down
these chemicals is greatly reduced [lack], allowing POPs to persist.
- Arctic animals, which store energy as fat for survival, are
particularly vulnerable to POP accumulation. Many of these animals are
also high on the food chain [carnivores], increasing their exposure
through biomagnification.

- Environmental Transport and Effects

41
- POPs can be transported over long distances by atmospheric currents,
from developed nations of the temperate zone up to remote areas like
the Arctic, where they are found in tissues of polar bears and seals.
- Toxicants can accumulate in water bodies like surface water,
groundwater, and through runoff and leaching of land drains into water
bodies, posing risks to human health when these water sources are
used for drinking water.

- Examples of POPs:
- DDT: A pesticide that is highly persistent and fat-soluble, leading to
bioaccumulation and biomagnification. DDT concentration increase
from plankton to fish to fish-eating birds.
- PCBs: Industrial chemicals that also persist in the environment and
accumulate in the food chain.
- Bt Toxin (in GM crops): Unlike DDT and PCBs, Bt toxin is not
persistent and breaks down more readily in the environment.

- Health and Ecological Impacts

- These can be passed up the food chain, leading to higher
concentrations in predators, which can cause severe health and
ecological impacts.
- Some toxicants degrade into simpler products that can still be
harmful. For example, DDT breaks down into DDE, which is also toxic.
- Temperature, moisture, sun exposure etc. affect the rate of
degradation.
- The high levels of POPs found in the breast milk of women in northern
regions highlight the severe impact of these chemicals on human
health, particularly for infants who rely on breast milk as their primary
food source.

42
EXTRA

- ‘The dirty dozen’ persistent organic pollutants (POPS)

43
DDT (1,1,1-trichloro-2,2-bis-(p-chlorophenyl) ethane)
- Initially used as a pesticide, DDT is toxic to a wide range of organisms,
not just the intended pests.
- Environmental Impact:
- DDT is introduced as a powder, emulsion, encapsulated, which is highly
persistent in the environment, with long-term presence in soil and
water.
- Biomagnification: DDT is lipid-soluble, leading to accumulation in the
fatty tissues of organisms and higher concentrations at each trophic
level.
- DDT has been detected in the blubber of Arctic ringed seals as early as
1970, despite being banned for decades in many countries.

- Pathway:
- DDT degrades into DDE via dehydrochlorination.
- Also, DDT undergoes reductive dechlorination to form DDD.
- DDT biochemical breakdown and removal
- Both DDE and DDD formed in the presence of NADPH and NADH, in
anaerobic conditions.
- Dechlorination is catalyzed enzymatically by cytochrome p450.
- Both DDT and DDE can be oxidized to form DDA, the primary excreted
product in animals.

- Toxic Effects of DDT

- Insects: DDT affects the peripheral nervous system, keeping sodium
ion channels open, causing random neuron firing, muscle twitching
(known as "DDT jitters"), and eventually death.
- Fish: Acts as a neurotoxin.
- Birds: Leads to eggshell thinning, which can reduce reproductive
success.
- Mammals:
- Localized liver and kidney damage.
- Neurotoxin effects.
- Teratogenic and carcinogenic effects.

44
- Biochemical Breakdown of DDT
- Detoxification:
- Insects: DDT-ase enzyme converts DDT to DDE, making it more water-
soluble.
- Mammals: DDT is dehalogenated to DDD and DDE, with DDD
accumulating in the liver [stored] and DDE being excreted in small
amounts, including through milk.
- Fish: DDT is highly resistant to metabolism, with very little excretion.

- Other Persistent Organic Pollutants (POPs)

- PCBs (Polychlorinated Biphenyls):
- Found in transformer fluids.
- Carcinogenic and mutagenic.
- Dioxins and Furans:
- Produced in high-temperature processes and herbicides.
- Highly carcinogenic, disturbing reproduction and suppressing immune
function.
- Hexachlorobenzene (HCB):
- Used in pesticides and produced during waste incineration and
metallurgical processes.
- Affects reproductive and immune functions.

- Effects of POPs on Living Organisms [how it kills]

- In Birds:
- Disrupts reproductive capabilities, including egg development and
mating behavior.
- In Mammals:
- Causes malformations in reproductive organs, infertility, and fewer
offspring.
- Acts as hormone disruptors by mimicking hormones, leading to various
physiological effects.
- Suppresses immune function, particularly in young animals.
- Likely carcinogenic.

45
3. Explain the action of few environmental toxicants’
epidemiology and animal testing
- Toxicology study effects in several major ways: wildlife toxicology
studies, human epidemiological studies, dose response studies in the
lab.
- Examples:
- In 1992 study data showed a significant decrease in men sperm’s count
over 50 years.
- Reason: Atrazine [endocrine disruptor]  chemical enter the
bloodstream and can mimic hormones. If the chemical bind to the sites
intended for hormone binding, can cause inappropriate response, thus
disrupting endocrine system. The hormone system is geared to work
with tiny concentrations of hormones; therefore, it can respond to tiny
concentrations of environmental contaminants.
- Studies also show that endocrine disruptors may cause the rise in
testicular cancer in many countries.

- Human epidemiological Studies:

- Involves the observation and analysis of individual patients. It's often
used in situations where exposure to a toxin is suspected to have
caused specific health effects.
- Advantages:
- These studies reflect real-world conditions, accounting for various
environmental, genetic, and lifestyle factors that could influence health.
- By including all real-life factors, these studies provide a broad
understanding of how environmental toxins affect health.
- Disadvantages:

46
- While epidemiological studies can identify associations between
exposure to toxins and health outcomes, they cannot definitively prove
causation.
- It may take many years to gather and analyze data, making these
studies resource-intensive and slow to produce results.

- Animal testing
- Determining Causes of Mortality in (Mass) Die-Off Events:
- Purpose: Investigates sudden, large-scale deaths in wildlife
populations to identify the cause, which could be due to toxins,
diseases or other environmental factors.
- Example: A mass die-off of birds may be linked to a toxic spill,
exposure to a harmful chemical, or an infectious disease.
- Laboratory Testing of Animals for Response to Toxicants:
- Purpose: Involves controlled experiments where animals are
exposed to specific toxicants in a lab setting to observe their
physiological, behavioral, or reproductive responses.
- Applications: Results can inform safety standards, environmental
regulations, and conservation strategies.
- Correlating Chemical Presence and Animal Presence in the
Field:
- Purpose: Field studies that examine the correlation between the
presence of specific chemicals in the environment and the
distribution, health, or population levels of wildlife.
- Approach: Researchers may measure toxin levels in soil, water, or
vegetation and assess their association with wildlife abundance or
health indicators.

4. Discuss the factors affecting environmental toxicity

47
- Individual Sensitivity: People are not equally sensitive to toxicants.
Sensitivity can vary based on factors such as:
- Sex: Biological differences between males and females can
influence how a toxicant is metabolized or its effects.
- Age: Babies, older adults, and individuals in poor health are
generally more sensitive to toxicants.
- Weight: Body mass can influence the concentration of a toxicant in
the body, with smaller individuals often being more affected by the
same dose.

- Type of Exposure:
- Acute Exposure: Involves a high level of exposure to a toxicant over a
short period.
- Can cause immediate and severe health effects, depending on the
substance and dose.
- Chronic Exposure:
- Definition: Involves lower levels of exposure to a toxicant over a long
period.
- Effects: Can lead to long-term health effects, such as cancer, organ
damage, or other chronic diseases.

Chapter 7

1. Explain about the toxicants in food

- Food safety is a critical concern, as various hazards can contaminate
food and pose risks to human health
- Physical Hazards: Physical hazards are foreign objects that can
accidentally enter food during production, processing, or handling.
Example, glass, stone, metal, wood.

48
- These hazards can cause injury, choking, or other physical harm if
ingested.
-
- Chemical Hazards: Natural Toxins. Examples:
- Mycotoxins: Such as Aflatoxins, produced by certain molds growing on
crops like grains, nuts, and fruits.
- Alkaloids: Found in certain plants, such as solanine in green potatoes,
which can cause gastrointestinal distress.
- Consumption of natural toxins can lead to acute poisoning, long-term
health effects, or even death.
-
- Residues: Pesticides and Veterinary Drugs.
- Chronic exposure to chemical residues can contribute to the
development of health issues like antibiotic resistance, hormonal
imbalances, and other toxic effects.
-
- Metals: Lead and mercury
- Heavy metals are toxic even at low concentrations and can lead to
neurological damage, developmental issues in children, and chronic
diseases.
- Toxins Formed During Food Processing:
- Acrylamide: Formed in starchy foods during high-temperature
cooking, such as frying or baking.
- Polycyclic Aromatic Hydrocarbons (PAHs): Produced when meat is
grilled or smoked at high temperatures.
- These processing-induced toxins are associated with increased risks of
cancer and other health problems.
-
- Microbiological Hazards: Bacteria, viruses, and parasites
- Bacteria:
 Salmonella: Common in raw poultry and eggs; causes foodborne illness
with symptoms like diarrhea and vomiting.
- Viruses:

49
 Hepatitis A: Spread through contaminated food or water, leading to
liver infection.
- Parasites:
 Toxoplasma gondii: Found in undercooked meat and contaminated
produce; can cause serious health issues, especially in pregnant women
and immunocompromised individuals.

2. Discuss the different natural toxins from food sources

- Natural toxins in foods: risks and types
- Natural toxins, found in both plants and animals, can pose significant
risks to human health, particularly when ingested over the long term.
- While not usually acutely toxic, these substances can lead to serious
health problems, including chronic illnesses, allergic reactions, and
other adverse effects.

- Risks of natural toxins in food:

- Can cause illness or, in rare cases, instant death. [general
toxicity]
- Carcinogenic
- Mutagenic
- Teratogenic:
- Endocrine Disrupters
- Microbial Pathogens

Categories of Natural Toxins in Foods [In detail below]

1. Endogenous Toxins of Plant Origin:

o Toxic Phenolic Substances:
 Flavonoids
 Tannins
 Coumarin
 Safrole
 Myristicin
o Cyanogenic Glycosides
50
o Glucosinolates
o Acetylcholinesterase Inhibitors
o Biogenic Amines
o Central Stimulants

2. Natural Contaminants:
o Mixing of Edible Plants with Toxic Plants
o Contamination from Toxic Substances Ingested by Animals
o Microbial Toxins

Endogenous Toxins of Plant Origin

Plants produce a variety of compounds that, while often beneficial or

harmless in moderate amounts, can act as toxins when consumed in large
quantities or by susceptible individuals.

1. Flavonoids

- Flavonoids are plant pigments widely present in human food,

particularly in fruits, vegetables, and some beverages like tea and
wine.

51
 - These compounds are polyhydroxy-2-phenylbenzo-γ-pyrone
derivatives, known for their antioxidant properties.
- A subgroup of flavonoids that includes yellow pigments like nobiletin,
tangeretin (found in citrus fruits), and 3,3ʹ,4ʹ,5,6,7,8-
heptamethoxyflavone (found in grapefruit).
- Location and Solubility:
- Flavones are typically found in the oil vesicles of fruit peels and are
apolar, meaning they are readily soluble in the oil.
- Toxicity and Health Effects:
- Mutagenicity: Flavones have been extensively studied for their
potential mutagenic effects (ability to cause mutations in DNA).
- Carcinogenicity: Among flavonoids, Quercetin is notable for being
the only one shown to be carcinogenic in mammals after oral
administration. Quercetin rich sources  apples, grapes, berries, citrus
fruits, broccoli, leafy greens, coffee, green tea.

Tannins

- Tannins, also known as tannic acids, are water-soluble polyphenols

found in a variety of plant foods, including tea, wine, and some fruits.
- Types of Tannins:  basis on degradation behavior and
botanical distribution
 Hydrolyzable Tannins: These are esters of gallic acid, digallic acid,
or ellagic acid with glucose or quinic acid. They can be broken down
(hydrolyzed) into simpler compounds.
 Condensed Tannins: These are polymers of flavonoids, specifically
leukoanthocyanidins, and do not easily break down.

- Toxicity and Health Effects:

- Tannins have been reported to cause acute liver injury, including liver
necrosis (cell death) and fatty liver disease. This toxicity is particularly

52
 concerning with excessive consumption of tannin-rich foods or
beverages.

Safrole

- Source: Found in oil of sassafras, black pepper, and certain other

plants.
- Toxic Effects:
- Safrole has been associated with the development of liver tumors in
rats.
- Due to its toxic potential, the use of safrole in food and beverages has
been prohibited in several countries, including the United States.

Coumarin

- Source: Found in cassis (blackcurrant), lavender, and lovage.

- Toxic Effects:
- Coumarin has been found to cause liver damage in animal studies,
particularly in rats.

Myristicin

- Source: Found in spices and herbs such as nutmeg, mace, black

pepper, carrot, parsley, celery, and dill.
- Toxic Effects:
- Nutmeg Toxicity:
- Myristicin can cause excitation of the central nervous system
(CNS).
- Toxic doses of myristicin can lead to tachycardia (rapid heart rate)
and failing salivation.
- Nutmeg has been abused as a narcotic due to its psychoactive effects
at high doses.

53
Cyanogenic Glycosides

- Source and Activation: Found in certain foods like cassava,

cyanogenic glycosides release cyanide when they undergo hydrolysis
through enzymatic activity.
- Toxic Effects:
- Consumption of cassava can lead to goiter because cyanide is
metabolized into thiocyanate by the enzyme rhodanase. [Thiocyanate
can inhibit iodine uptake by the thyroid gland, leading to iodine
deficiency and goiter]
- Detoxification Methods:
- Peeling, Washing, Cooking, and Fermenting: These processes can
remove or volatilize cyanide from foods, reducing their toxicity.

Goitrogens (Glucosinolates)

- Source: Found in many commonly consumed plants such as cabbage,

cauliflower, Brussels sprouts, broccoli, turnip, radish, and oil
seed meals.
- They can be considered as natural toxins, but also as antinutritive.
These compounds can inhibit iodine uptake by the thyroid gland,
leading to iodine deficiency.
- Toxic Effects:
- Glucosinolates themselves are not directly toxic, but their hydrolysis
products, including isothiocyanates and nitriles, are active agents.
Activation:
 The enzyme responsible for glucosinolate hydrolysis becomes active
when plant cells are damaged through cutting or chewing, leading
to the release of toxic compounds.
- Some isothiocyanates have been shown to be embryotoxic in rats
and cytotoxic and mutagenic in vitro.

54
- Some nitriles are precursors to N-nitroso compounds, which are
known carcinogens.

Acetylcholinesterase Inhibitors

- Source: Found in several edible fruits and vegetables, particularly in

the Solanaceae family, including potatoes, eggplants, and
tomatoes. The most active alkaloid in this group is solanine.
- Toxic Effects:
- Gastrointestinal and Neurological Symptoms: Oral ingestion of
solanine can lead to symptoms like nausea, vomiting, diarrhea, and
neurological disturbances such as headaches and confusion.
- Potato Poisoning: Potatoes, especially when green or sprouting, can
contain lethal amounts of solanine. The most potent inhibitors are
found in these conditions, and the symptoms of poisoning may be due
to the combined actions of solanine with other glycoalkaloids like
chaconine and tomatine.
- Factors Affecting Toxicity:
- Solanine levels vary based on the degree of potato maturity at harvest,
nitrogen fertilization rates, storage conditions, and exposure to light
(which causes greening).
- Most of the solanine is found in the potato skin, so peeling can remove
much of the toxin, though solanine is heat-stable and insoluble in
water, meaning toxic potatoes cannot be rendered harmless by
cooking.
- Safety Limits:

55
- 20 mg of solanine per 100 g fresh weight is considered the upper
safety limit for consumption.

Biogenic Amines

- Biogenic amines (BAs) are low molecular weight organic bases that
play significant roles in various biological processes but can also pose
health risks when present in high concentrations in food.
- Sources: naturally found in animals, plants, and microorganisms.
Common sources include fermented foods (such as cheese and
fermented vegetables), vegetables, and meats. They are particularly
high concentrated in fish and cheese.

- Formation:
- BAs are primarily formed through the enzymatic decarboxylation of
amino acids by microorganisms.
- Factors Influencing Formation:
- Microorganisms: Several bacteria such as enterococci, lactobacilli,
enterobacteriaceae, and pediococci are known to produce these
amines.
- pH
- Temperature
- Salt Concentration (Cheese)
- Availability of Free Amino Acids
- Level of Starter Culture (Cheese)
- Legal Limits and Toxicity
- General Safety: A total of 1000 mg/kg of BAs (based on histamine
intoxication and amine concentration in food) is considered dangerous
for human health.
- The toxic dose of BAs varies depending on individual sensitivity and
specific characteristics of the amine.
- Biological Activity
- Cell Proliferation

56
- Regulation of Nucleic Acid Function
- Protein Synthesis
- Toxic Effects:
- Histamine Intoxication
- Nausea
- Respiratory Distress
- Hot Flushes
- Heart Palpitation
- Headache
- Hypertension & Hypotension

Mushroom toxins

- These species are known for their high content of amatoxins, potent
compounds responsible for severe poisoning cases.
- Species Identified:
- Amanita bisporigera
- Amanita temifolia
- Amanita ocreata
- Amanita suballiacea
- Galerina autumnalis
- Lepiota brunneolilacea

- Four Categories of Mushroom Toxins

1. Neurotoxins:
o Neurological symptoms include profuse sweating,
hallucinations, depression, spastic colon, excitement,
convulsions, and coma.
2. Protoplasmic Poisons:

57
 o These toxins cause generalized destruction of cells, leading to
organ failure.
 Amatoxins fall into this category, leading to severe liver and
kidney damage.
3. Gastrointestinal Irritants:
o These toxins produce rapid, transient nausea, abdominal
cramping, vomiting, and diarrhea.
 While they may cause significant discomfort, they are less
likely to result in long-term damage compared to
protoplasmic poisons.
4. Disulfiram-like Toxins:
o These usually nontoxic compounds produce no symptoms on
their own.
o However, if alcohol is consumed within 72 hours of ingestion,
they can cause vomiting, nausea, headache, flushing, and
cardiovascular disturbances.

Phases of Mushroom Poisoning

 Phase 1:
o Symptoms typically start 6 to 24 hours post-ingestion.
 This period may be asymptomatic, leading to delayed
diagnosis.
 Phase 2 (Gastrointestinal Phase):
o Severe vomiting, abdominal cramps, nausea, and watery
diarrhea.
 Phase 3:
o Lasts about 12-24 hours
o Characterized by improved clinical symptoms but also the
beginning of liver necrosis

58
 Phase 4:
o This phase is marked by hepatic failure, encephalopathy (brain
dysfunction due to liver failure), internal bleeding, and potentially
acute renal failure.
o Without treatment, these complications can lead to death.

3. Explain the different natural contaminants of food sources

- Natural contaminants can also originate from biological systems
different from those in which they occur.
• There are three important sources:
- Raw materials of plant origin can become contaminated if they are
mixed with toxic non-nutritive plant species.
- Raw materials of animal origin, mainly fish and milk, can also
become contaminated if the animal has ingested toxic substances of
natural origin.
- Contaminants of natural origin can be the products of
microorganisms.

4. Describe with names examples

- Mixing of edible plants with toxic plants
- Example: cereal contamination
- Causative agents: Pyrrolizidine alkaloids produced by the genera
Senecio, Crotalaria, and Heliotropium.
- Cause: acute liver damage and vein lesions, liver cancer

- Natural toxins in aquatic organisms [shellfish poisoning]

- Paralytic shellfish poisoning (PSP) consumption of shellfish that
have become contaminated with a group of toxins from the ingestion
of toxic plankton, in particular toxic dinoflagellates.
- The dinoflagellates produce a complex mixture of toxins. One of
the components has been identified as saxitoxin.
 The shellfish involved are pelecypods, a family of mollusks.

59
  Symptoms  tingling and burning in face, lips, tongue, and
ultimately the whole body, and paresthesia followed by numbness,
general motor incoordination, confusion, and headache.
 Develop within 30 minutes after ingestion. Death, preceded by
respiratory paralysis, occurs within 12 hours.
 Chance of contamination and poisoning is highest during red tide
(dinoflagellate bloom*)
 Ordinary cooking destroys up to 70% of the toxin(s) and pan- frying
destroys even more, but there may still be sufficient toxin left in the
mollusks to cause serious poisoning.

- Contaminants of natural origin [Mycotoxin]

- Sources:
- Mycotoxins [fungal secondary metabolites] are typically produced by
fungi from the genera Aspergillus, Penicillium, and Fusarium.
- Health Effects:
- Acute Toxicity
- Chronic Toxicity: such as cancer, liver cirrhosis, and reproductive
issues.
- Toxic syndromes resulting from the consumption of mycotoxin-
contaminated food or feed are referred to as mycotoxicosis.
- Environmental Influence:
- The contamination of food and feed with mycotoxins is heavily
dependent on the environmental conditions that promote mold
growth and toxin production.
- Not all foods with visible mold contain mycotoxins, and conversely,
the absence of mold does not guarantee that mycotoxins are not
present.
- Mycotoxins can persist even after the molds have been killed during
food processing, due to their chemical stability.
- Aflatoxins: A Notable Mycotoxin Group
- Aflatoxins are primarily produced by Aspergillus species, specifically
A. flavus and A. parasiticus.
- These molds thrive in high temperatures and humid conditions.

60
 - Health Risks:
- Aflatoxins are potent carcinogens and have been linked to liver
cancer and cirrhosis.
- They are particularly dangerous because there is no known safe
threshold for exposure; even small amounts can be harmful.
- Commonly Contaminated Foods:
- Aflatoxins can be found in a variety of foods, including tree nuts,
peanuts, corn, and cottonseed oil.
- The major aflatoxins of concern are B1, B2, G1, and G2, with
aflatoxin B1 being the most toxic and carcinogenic.
- Detection:
- Aflatoxins can be detected using Thin Layer Chromatography (TLC),
where aflatoxins B1 and B2 fluoresce blue and G1 and G2 fluoresce
green under UV light.
- Metabolism and Excretion:
- Aflatoxin B1 is metabolized into aflatoxin M1, which is excreted in
the milk of dairy cattle that have consumed contaminated feed. This
can lead to the presence of aflatoxins in milk products.
- Toxicity:
- The lethal dose (LD50) of aflatoxins can range from 0.5 to 10 mg/kg
body weight, varying among different animal species.

Chapter 8
1. Discuss the role of toxic chemicals, rodenticide, heavy metals in
toxicity
Rodenticide: Fluoroacetate:
- Found in South African plants, used as a deterrent for herbivores.

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- Not inherently toxic but metabolized into fluorocitrate [suicide
substrate], which inhibits the citric acid (TCA) cycle, reducing cellular
respiration.
- Fluorocitrate competes with citrate for aconitase, leading to
energy deficiency and deficiency of toxic intermediates [alpha
ketoglutarate].
- Like acetyl CoA it takes part in the citrate synthase reaction.

 Effects: Hyperammonaemia = nausea → convulsions → coma → death

 Vomiting, abdominal pain, tachycardia, renal failure, muscle spasms,
CNS symptoms.

Metallic poisons
Cadmium Toxicity:
- Source: Found in alloys, paints, plastics, cigarette smoke, and certain
industrial applications.
- It is poorly absorbed from the gut (5-8%).
- Half-life in body is 7-30 years, highly toxic to mammals.
- Inhaling Cd metal and taking in Cd++ produces toxicity.

- Mechanism:

62
- Cadmium (Cd++) [similar to Zn ++] binds to sulfhydryl (-SH) groups,
inactivating enzymes, inducing oxidative stress, and causing damage to
various organs, including kidneys, lungs, heart, bones, and
reproductive organs.

- Effects: Leads to conditions like cardiovascular damage, testicular

damage, kidney damage, lung damage, osteomalacia and brittle bones,
carcinogenic leading to testicular cancer.

Kidney Damage

- The release of cadmium (Cd) into tubular cells leads to the

degeneration of the proximal convoluted tubule in the kidney.
- This process generates reactive oxygen species, which disrupt Na+/K+
ATPase enzymes critical for maintaining cellular ion balance.
- The resulting oxidative stress and enzyme disruption cause the
depolarization of microtubules, leading to structural instability within
the cells.

63
- These damaging effects culminate in irreversible nephrotoxicity,
ultimately resulting in renal failure.
Testicular Damage
- Cadmium exposure is also linked to reproductive toxicity, including
testicular necrosis and decreased sperm production.

Lung Damage
- Occupational exposure to cadmium dust or fumes can lead to lung
damage.
- Acute exposure causes chemical pneumonitis, characterized by
inflammation of lung tissue.
- Chronic exposure leads to pulmonary fibrosis, a condition where lung
tissue becomes scarred, reducing lung function.

Bone Damage:
- Cadmium exposure is historically associated with Itai-itai disease, a
painful condition caused by severe bone demineralization and fractures.

64
- This disease was first identified in Japan among people who consumed
rice irrigated with cadmium-contaminated water.
- Cadmium interferes with calcium metabolism, leading to bone loss
(osteoporosis) and increased fracture risk.

Cardiovascular Damage
- Cadmium (Cd) acts as a cardio depressant, impacting both the
mechanical and electrical functions of the heart.
- Mechanically, cadmium antagonizes calcium's role in myocardial
contraction, impairing the heart's ability to contract effectively.
- Electrically, cadmium disrupts calcium's electrical activity, leading to
prolonged conduction intervals.
- cadmium is 100 times more potent as an electrical disruptor than as a
mechanical disruptor.

Mercury Toxicity:
- Forms: Elemental mercury (Hg), inorganic mercury (Hg++, Hg+), and
organic mercury (methylmercury).
- All forms are toxic and can cross the placenta to the fetus.
- Elemental Mercury
- When mercury (Hg) is in vapor form, it is completely absorbed by the
lungs upon inhalation.
- After absorption, it easily crosses the blood-brain barrier (BBB), allowing
it to affect the central nervous system (CNS).
- Ingested mercury is not absorbed by the gastrointestinal tract and is
therefore relatively harmless.

65
- Mercury exposure typically occurs in occupational settings where
workers might inhale the vapor.

- Inorganic Mercury
- Mercuric (Hg++) and mercurous (Hg+) ions are poorly absorbed by the
gastrointestinal (GI) tract, but when absorbed, they can cause
significant damage, including diarrhea, ulceration, and
necrosis.
- Once absorbed, mercury tends to accumulate in the kidneys, where it
induces toxicity through two primary mechanisms:
o Stimulation of the immune system, particularly affecting the
glomerulus,
o Causing lipid peroxidation and oxidative stress.
- This can lead to renal failure, which may occur as quickly as 24 hours
after exposure, with the cells of the proximal convoluted tubule being
the primary targets.
- Sulphate-reducing bacteria (SRB) can convert inorganic mercury into
organic mercury compounds, which are more toxic.

- Organic Hg on CNS
- The main site of action for organic mercury is the central nervous
system (CNS).
- Organic mercury generates reactive oxygen species, leading to
oxidative stress.
- Neurons with low glutathione activity are particularly vulnerable to this
oxidative damage.
- Exposure to organic mercury can cause a range of neurological effects,
including paresthesia, paralysis, movement defects, deafness,
blindness, and insanity.

- Different forms of mercury affect various parts of neurons in

distinct ways:

66
- Methyl Mercury: Due to its lipophilicity, methyl mercury primarily
affects the myelin sheath, leading to decreased neuronal excitability.
- Divalent Mercury: This form interferes with calcium signaling, which
disrupts cholinergic processes essential for neuronal communication
and function.
- Placental Transfer: Organic mercury can cross the placenta,
impacting the developing nervous system of the fetus. Mercury also
disrupts microtubule polymerization, which can lead to defects in
neuronal migration and development.

- Mechanism of mercury toxicity: High affinity for SH groups in

enzymes, leading to inhibition of critical enzyme systems and disruption
of cellular functions.

- Enzyme Systems Inhibited

- Cadmium toxicity inhibits several critical enzyme systems, including
membrane ATPase, which disrupts cellular energy balance.
- It impairs brain pyruvate metabolism and lactate dehydrogenase
activity, hindering energy production.
- The synthesis of fatty acids is also affected, leading to metabolic
imbalances.
- These disruptions in active transport and mitochondrial metabolism
cause significant changes in membrane permeability and transport
processes, ultimately resulting in cell death.

Lead Toxicity:
- Lead toxicity is caused by both elemental lead (Pb) and its ionic form
(Pb++), with organ lead compounds being highly toxic.
- Source:
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- industrial activities such as smelting, rubber manufacturing, tinning,
and pottery production.
- Lead pipes, especially in soft-water areas, old lead-based paint,
imported lead-contaminated toys, and glazed pottery.
- Exhaust fumes from the combustion of petrol containing tetraethyl lead,
an antiknock agent, can lead to exposure to lead. Although the amount
of lead inhaled from exhaust fumes may be less than that found in food,
it is more readily absorbed through the lungs. This exposure can result
in lead-induced anemia.
 Anaemia
 Lead toxicity causes anemia by inhibiting several steps in the heme
biosynthetic pathway.
 One key enzyme affected is δ-aminolevulinic acid dehydratase,
which is responsible for converting ALA (δ-aminolevulinic acid) to
porphobilinogen.
 When this enzyme is inhibited, ALA accumulates and is excreted in
the urine.
 ALA also inhibits the release of GABA and competes for receptor
sites in the nervous system.
 Lead inhibits ferrochelatase, the enzyme necessary for incorporating
Fe²⁺ into protoporphyrin IX to produce heme.
 Instead of iron, protoporphyrin is incorporated into red blood cells,
leading to the chelation of Zn²⁺ as the cells circulate through the
body.
 The presence of Zn-protoporphyrin, which fluoresces under specific
light, serves as a diagnostic marker for lead-induced anemia.

68
- Effects:
- Kidneys:
- Affects the proximal tubules.
- Inhibits absorption mechanisms, leading to amino aciduria and
glycosuria.
- Gastrointestinal Tract:
- Causes constipation, colic, and vomiting.
- May result in liver damage.
- Sex Organs:
- Decreases reproductive function, particularly in males with occupational
exposure.
- Leads to reduced testosterone levels.
- Bones:
- Lead is deposited in hydroxyapatite, acting as a "sink" for lead.
- Unlike cadmium, lead is not osteotoxic.

69
- Bone deposition provides a means of assessing lead exposure.
- May adversely affect fetal and postnatal bone development.
- Neuromuscular System:
- Causes muscle paralysis, wasting, and motor neuron damage.
- Neurotoxic effects of Lead:
- Lead readily crosses the blood-brain barrier (BBB) via the action of Ca-
ATPase.
- Its neurotoxic effects manifest as encephalopathy and peripheral
neuropathy.
 Encephalopathy: Accompanied by cerebral edema. Leads to
degeneration of neurons, particularly in the cerebral cortex.
 Peripheral Neuropathy: Involves the degeneration
(demyelination) of peripheral motor nerves.

- In neurons, lead alters the release of Ca²⁺.

- It causes the opening of pores in mitochondria, leading to apoptosis.
- Lead interferes with the timing of cell-to-cell communication in
developing neurons.
- Disrupts Ca²⁺-dependent neurotransmitter release, affecting
neurotransmitters like acetylcholine and dopamine.
- Lead also impacts non-neuronal cells, including oligodendroglia
(involved in myelination) and astroglia (regulating the external
environment).
- Neurotoxicity results from both direct effects on neural transmission
and indirect effects caused by edema and cellular hypoxia.
- Chronic exposure to lead can result in a variety of effects on the central
nervous system (CNS) → Headache and fatigue, Memory loss in adults,
Mental retardation in children, Convulsions, Cerebral palsy, and
Blindness.

Arsenic Toxicity:

70
- Arsenic is not strictly classified as a heavy metal but is generally
grouped with metal toxicants.
- Arsenic exists in two forms: pentavalent (AsO₄³⁻) and trivalent (AsO₂⁻).
- These forms of arsenic inhibit different stages of carbohydrate
metabolism.

- Pentavalent arsenic (AsO₄³⁻) is a homologue of phosphate.

- It inhibits the enzyme glyceraldehyde 3-phosphate dehydrogenase.
- This inhibition leads to the formation of 1-arseno-3-phosphoglycerate.
- The end-products are AsO₄³⁻ and 3-phosphoglycerate.
- This process causes a bypass of the phosphoglycerate kinase reaction.
- Consequently, there is a reduction in ATP production, which is essential
for energy production.

71
- Arsenite, most toxic form of arsenic → inhibits enzymes with lipoic acid
as cofactor e.g. Pyruvate dehydrogenase, a-ketoglutarate
dehydrogenase.

- Effects and Treatment of Arsenic Poisoning

- Arsenic leads to the accumulation of pyruvate and consequently
lactate.
- Decreased energy production and the development of acidosis both
contribute to neurological damage and death.
- Treatment of arsenic poisoning involves using compounds that compete
with lipoic acid, such as 2,3-dimercaptopropanol.

2. Describe the mechanism driven by the metals for causing

toxicity in major system
- Blockage of Metabolic Pathways
- Relative or total blockage of one or more metabolic pathways may
result from the inhibition of enzymes or damage to structural
organizations, such as the mitochondrion.
- A major cause of cell death is the loss of ATP supply due to the
inhibition of oxidative phosphorylation.

- Mechanisms of Inhibition of ATP Supply

- Reduced oxygen delivery to tissues, caused by substances like carbon
monoxide (CO) or nitrite, can impair cellular respiration.
- Additionally, the inhibition of carbohydrate metabolism by agents such
as fluoroacetate disrupts the energy production process.
- The electron transport chain may be inhibited by compounds like
rotenone, antimycin A, cyanide, CO, and azide.

72
- Uncoupling of oxidative phosphorylation can be induced by agents such
as nitrophenol and organotins, leading to inefficient ATP synthesis.
- Oligomycin specifically inhibits ATP formation.
- Structural damage to the mitochondrion also compromises ATP supply.
3. Explain the toxic chemicals, chemical warfare agents and
their effects
- Categories of Hazardous Substances
- Toxics: These are poisons that usually cause systemic damage to the
body.
- Reactive: These substances chemically react with everyday materials.
- Corrosives: Corrosives are substances that can corrode materials.
- Flammables: Flammable substances are capable of burning easily.
- Compressed Gases: These are gases stored under very high pressure
in cylinders. They may or may not contain toxic or flammable gases.

- Hazardous Chemicals
- A hazardous chemical is defined as any chemical that fits into one or
more of several categories. These include physical hazards, health
hazards, simple asphyxiants, combustible dust, pyrophoric gases, and
hazards not otherwise classified.
- Physical Hazards
- Physical hazards refer to chemicals that can pose risks such as fire,
explosion, or violent reactions.
- Health Hazards
- Health hazards involve chemicals that are detrimental to health,
potentially causing both short-term (acute) and long-term (chronic)
health problems.
- According to OSHA, a health hazard is classified as any chemical that is
toxic, corrosive to the skin or eyes, a respiratory sensitizer, or has the
potential to cause cancer, birth defects, or reproductive issues.
Additionally, chemicals that attack specific organs or are harmful or
deadly when inhaled are also considered health hazards.
- Gases

73
- Industrial Toxic Thresholds
- By Inhalation
- TLV/TWA (Threshold Limit Value-Time Weighted Average): This
value represents the average concentration of a chemical to which a
worker can be exposed day-to-day without experiencing adverse health
effects.
- STEL (Short Term Exposure Limit): This limit allows for exposure to
higher concentrations than the TLV/TWA for up to 15 minutes at a time,
but such exposure should not occur more than four times per day.
- TLV-C (Threshold Limit Value-Ceiling): This threshold represents a
concentration that should never be exceeded at any time, according to
ACGIH.
- REL (Recommended Exposure Limit): Established by the National
Institute for Occupational Safety and Health (NIOSH), this is a
recommended exposure limit for chemicals.
- PEL (Permissible Exposure Limit): This limit, set by the
Occupational Safety and Health Administration (OSHA), dictates the
maximum concentration of a chemical to which workers may be
exposed.
- IDLH (Immediately Dangerous to Life or Health): This
concentration, defined by NIOSH, indicates levels of a chemical that
pose an immediate threat to life or health.
- LC50 (Lethal Concentration 50%): This value represents the
concentration of a chemical in the air that is lethal to 50% of test
animals. A concentration less than 200 mg/kg is considered highly toxic.
74
- By ingestion: This measure indicates the dose of a chemical ingested
or absorbed through the skin that is lethal to 50% of test animals. A
dose less than 200 mg/kg is considered highly toxic.
- Carcinogens: These are chemicals known to cause cancer. Exposure
levels are often not applicable or are set at levels that minimize risk,
depending on the substance.

Relative Toxicity by Inhalation

- >500 ppm: Toxic
- 101-500 ppm: Moderately Toxic
- 1-100 ppm: Highly Toxic
- <1 ppm: Super Toxic

Chemical Warfare Agents

- These are designed to harm or kill through chemical reactions.
- Types of Chemical Weapon Agents
- Nerve Agents:
o GA: Tabun
o GB: Sarin
75
 o GD: Soman
o GF: (specific agent not listed)
o VX: Methylphosphonothioic acid
- Blister Agents:
o HD: Sulphur mustard (Yperite)
o HN: Nitrogen mustard
o L: Lewisite
o CX: Phosgene oximine
- Choking Agents:
o CG: Phosgene
o DP: Diphosgene
o Cl: Chlorine
Types of Biological Weapon Agents
- Anthrax
- Botulinum Toxins
- Brucellosis
- Cholera
- Clostridium Perfringens Toxins
- Congo-Crimean Hemorrhagic Fever
- Ebola Hemorrhagic Fever
- Melioidosis
- Plague
- Q Fever
- Ricin
- Rift Valley Fever
- Saxitoxin
- Smallpox
- Staphylococcal Enterotoxin B
- Trichothecene Mycotoxins
- Tularemia
- Venezuelan Equine Encephalitis

- Blistering Agents

76
- Blistering agents cause severe tissue damage and are known for their
persistent effects.
- These agents covalently bond to nucleic acids, proteins, and
nucleotides, leading to extensive and prolonged harm to affected
tissues.

- Choking Agents
- Choking agents attack lung tissue and inhibit essential enzymes,
leading to the production of hydrochloric acid (HCl) in the lungs.
- They also increase the permeability of alveoli, which impairs respiratory
function and causes severe respiratory distress.

- Psychotomimetric Agents
- Psychotomimetric agents affect the brain, leading to an inability to
make decisions, altered vision, and hallucinations.
- They cause disorientation and can have long-lasting effects on mental
functioning.

77
- Blood Agents
- Blood agents are colorless gases that are absorbed into the blood
through breathing.
- They bind with metal-containing enzymes, inhibiting oxidative
processes within cells and disrupting essential cellular functions.

- Nerve Agents
- Nerve agents poison the nervous system and are characterized by their
high toxicity levels.
- They produce immediate effects by inhibiting the actions of
acetylcholinesterase, which disrupts normal muscular contractions and
nerve signal transmission.

- Nerve Agents
- Sarin Gas:
- Sarin is a highly toxic nerve agent that over-stimulates muscles and
vital organs. It can be inhaled or absorbed through the skin.
- As little as 100 mg can be fatal within a few minutes.
- Sarin is 500 times more toxic than cyanide, making it extremely
dangerous.
- It is also expensive and unstable, adding to its risks.
- VX Nerve Gas:
78
- VX is the deadliest nerve agent in the U.S. arsenal.
- Even a fraction of a drop absorbed through the skin is lethal.
- It is complicated and dangerous to produce, being 100 times more
deadly than sarin gas when absorbed and twice as deadly when
inhaled.
- Novichok (Soviet V-gas):
- Novichok is estimated to be five times more powerful than VX.
- According to Lev Fedorov and Vil Mirzayanov in 1992, it is described as
a "new toxic agent" with injuries that are "practically incurable," and
those affected may remain disabled for life.
- Novichok is also notably difficult to detect.

Chapter 9
1. Explain about genotoxicity and teratogenicity with at least one
example
- Genetic Toxicology
- Genetic toxicology involves assessing the harmful effects of chemicals
or physical agents on the hereditary material and related genetic
processes in living cells.
- Damage to DNA can occur at the gene or chromosomal level,
potentially leading to heritable mutations that result in genetic
disorders, congenital defects, or cancer.
- The primary targets of DNA damage are:
- Somatic Cells: Damage to these cells is detrimental to the exposed
individual.

79
- Germinal Cells: Damage to these cells can lead to heritable effects that
may be passed onto offspring.
- Mitochondria: Damage to mitochondrial DNA affects the exposed
individual and can also be passed to progeny via maternal inheritance.
- Genotoxic Classification Scheme

- Mechanisms for Genetic Damage

- Genotoxic chemicals can produce genetic damage even at subtoxic
levels.
- The types of DNA damage include:
- Single- and Double-Strand Breaks
- Crosslinks
- Chemical Additions (Adducts)
- Furthermore, DNA replication can introduce errors, such as incorrect
base substitutions, and this process may be worsened by certain
genotoxic agents.

Types of Genetic Damage

1. Base Substitution: This occurs when the correct nucleotide is
replaced by an incorrect one.
o Transition: A purine is replaced by another purine, or a pyrimidine
is replaced by another pyrimidine.
o Transversion: A purine is replaced by a pyrimidine, or a pyrimidine
is replaced by a purine.

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2. Frameshift Mutation: This involves the addition or deletion of one or
a few base pairs (not in multiples of three, which are codons) in protein-
coding regions, leading to a shift in the reading frame of the DNA
sequence.
3. Structural Chromosome Aberrations:
o For non-radiomimetic chemicals, these aberrations can arise from
errors in DNA replication on a damaged template.
o Radiomimetic chemicals can directly induce strand breaks,
contributing to structural chromosome aberrations.
4. Numerical Chromosome Changes:
o Numerical Aberrations: These involve deviations from the normal
diploid chromosome number in the nucleus.
o Monosomies and Trisomies: These arise from errors in
chromosome segregation, where there is a loss (monosomy) or
gain (trisomy) of one or more chromosomes.
o Aneuploidy: This refers to a numerical deviation from the modal
chromosome number and can result from chemical effects on
tubulin polymerization or spindle microtubule stability.
5. Sister Chromatid Exchanges (SCE):
o These exchanges can occur during the S phase of the cell cycle as a
consequence of replication errors. They are reciprocal exchanges
between sister chromatids.

2. Discuss the classes and effects of genotoxic chemicals and

thalidomide classes of Genotoxic Chemicals:
Genotoxic Chemicals
1. Probable Non-Genotoxic (Non-Mutagenic) Carcinogens:

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- These chemicals are considered tumor promoters or are negative for
genotoxicity in vivo.
- They are less likely to cause genetic damage directly.
2. Questionable Carcinogens:
- These chemicals have uncertain carcinogenic potential and may require
further investigation to determine their genotoxicity.
3. Probable Genotoxic Carcinogens:
- These chemicals are likely to cause genetic damage and have a higher
potential to be carcinogenic.
4. Mode of Carcinogenic Action Unknown:
- For these chemicals, the mode of action is not fully understood, and
their genotoxicity in vivo is unknown or unclear.
- In terms of priorities, chemicals in categories 2, 3, and 4 are considered
most important for in vitro mammalian cell tests to detect potential
genotoxicity.

2-Mercaptobenzothiazole
- 2-Mercaptobenzothiazole shows some evidence of carcinogenic activity
in male F344/N rats, with tumors including mononuclear cell leukemia,

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 pancreatic acinar cell adenomas, adrenal gland pheochromocytomas,
and preputial gland adenomas or carcinomas.
- In female F344/N rats, it is associated with adrenal gland
pheochromocytomas and pituitary gland adenomas.
- In contrast, there is no evidence of carcinogenic activity in male B6C3F1
mice, but there is equivocal evidence in female B6C3F1 mice, with
potential hepatocellular adenomas or carcinomas.
- The chemical does not produce DNA adducts and is negative for
micronucleus (MN) formation in vivo.
- Given these mixed results, 2-mercaptobenzothiazole may need to be
re-tested to confirm its carcinogenic potential.

Furfural
- Furfural is considered one of the more contentious chemicals among
those purported to be non-genotoxic.
- In the micronucleus assay (MLA) conducted by the National Toxicology
Program (NTP), the lowest positive concentration observed was 200
µg/ml with S9 mix and 400 µg/ml without S9 mix (2.1 or 4.2 mM).
However, treatments in this assay were only administered for 3 hours.
- Similarly, in the chromosomal aberration (CA) test, the lowest positive
concentration observed was 300-400 µg/ml (3-4 mM). It is important to
note that the NTP protocols for these tests involved short treatment
durations and early sampling times, which may impact the results.

Thalidomide
- Thalidomide was initially used as a mild sedative to alleviate morning
sickness during pregnancy.

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- However, it was found to be teratogenic, meaning it could induce
malformations during organogenesis.
- When taken between days 24 and 33 of pregnancy, thalidomide caused
a range of severe effects, including:
 Phocomelia: Malformation of the limbs, where the limbs are
severely shortened.
 Amelia: Absence of limbs.
 Defects in Eyes and Ears: Various abnormalities affecting
these organs.

3. Explain the mechanism of thalidomide causing teratogenicity

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- Mechanisms for the Embryotoxicity of Thalidomide
- Formation of Toxic Metabolites: Thalidomide is metabolized into
compounds that exhibit toxicity in rats and mice.
- Accumulation in Embryonic Tissues: Thalidomide tends to accumulate in
embryonic tissues, where it can cause developmental damage.
- Intercalation into DNA: The drug can insert itself into the DNA,
potentially disrupting normal cellular processes and leading to genetic
damage.
- Inhibition of Angiogenesis: Thalidomide interferes with the formation of
new blood vessels, which is crucial for normal fetal development.
- Inhibition of Genes Coding for Integrins: Thalidomide inhibits genes that
encode integrins, proteins essential for cell adhesion and tissue
development.
- Inhibition of Growth Factors: The drug affects the actions of various
growth factors that are important for normal cellular growth and
development.

- Mechanism of Action of Thalidomide

- Thalidomide acts by intercalating into specific poly G (GGGCGG) sites
within the promoter regions of certain genes.
- Thalidomide specifically binds to poly G sequences in the promoter
regions of genes.
- Only the S-form of thalidomide fits into the major groove of the DNA,
allowing it to bind effectively to poly G sequences.
- The R-form of thalidomide cannot bind due to steric hindrance,
preventing its interaction with the DNA.
- The promoters for integrin subunits (such as av and b3), insulin-like
growth factor 1 (IGF-1), and fibroblast growth factor 2 (FGF-2) contain
poly G sites and are targeted by thalidomide.

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- Current Uses of Thalidomide
- Skin Conditions in Leprosy:
- It is effective in managing skin complications associated with leprosy.
- AIDS:
- Thalidomide is used in the treatment of certain complications related to
AIDS.
- Cancer:
- It is employed in the treatment of some cancers, including multiple
myeloma and certain types of lymphoma.
- Anti-Rheumatic Drug:
- Thalidomide is used to treat specific inflammatory conditions, such as
erythema nodosum leprosum, a complication of leprosy.
- Fetal Alcohol Syndrome (FAS):
o Fetal Alcohol Syndrome is a condition caused by alcohol exposure
during pregnancy. It is characterized by:
 Infants with FAS often exhibit slowed growth both prenatally and
postnatally.

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  Distinctive facial abnormalities can include a smooth philtrum,
thin upper lip, and small eye openings.
 Individuals may experience a range of neurological issues,
including cognitive impairments, developmental delays, and
behavioral problems.

4. Discuss the treatment and prevention of teratogenicity in

general
 Avoid exposure to known teratogens during pregnancy. Use
alternative medications that are safe for pregnant women.
 Once teratogenic effects occur, treatment options are limited. Focus
is on managing and supporting developmental defects.
 Educate pregnant women about potential teratogens and monitor for
early signs of exposure.

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