Natural bioactive compounds that trigger apoptosis in medicinal plants might be a promising new s... more Natural bioactive compounds that trigger apoptosis in medicinal plants might be a promising new source of anti-cancer medications. The present study aimed to assess the anti-cancer potential of the powerful therapeutic plant Chrysanthemum indicum. The whole plant extract was extracted in n-hexane and ethanol solvents to test anticancer and antioxidative capabilities on the cancer HepG2 cell line. The cells in the untreated group had not been exposed to any of the plant extracts. Ethanol and n-hexane extracts were administered to the treated cell lines. Cell viability was estimated by employing standard methodologies for the anticancer potential. Cell death and viability were measured by MTT, trypan blue, and crystal violet tests. The rate of apoptosis was measured in all groups using an Annexin-V ELISA. Furthermore, antioxidant enzyme activity studies were carried out to investigate the scavenging activities of nitric oxide, DPHH radicals, H2O2, and superoxide, scavenging power of t...
International Journal of Applied and Experimental Biology
Azadirachta indica constituents exhibit potential therapeutic role to control cancer progression.... more Azadirachta indica constituents exhibit potential therapeutic role to control cancer progression. The potential anti-cancerous activity of neem extracts reduces cell propagation, migration, inflammation, and invasion. The current review describes the importance and effectiveness of neem extracts in different solvents against various cancers. Studies performed in different geographic locations from 2001 to 2021 were searched in Google, PubMed, Google Scholar, and NCBI. The relevant information has been retrieved, including the type of extracts, solvents, year of publication, and type of cancers. A total of 125 relevant studies published in different journals have been screened, among which the most commonly used neem part was leaf (n = 79), followed by seed (n = 18), flower (n = 10), oil (n = 10), bark (n = 10), roots (n = 6), fruit (n = 6), gum (n = 5), limonoids (n = 5), nimbolide (n = 4), stem (n = 4), xylem (n = 1), and as herb (n = 1). Similarly, the most commonly investigated c...
pncB1 and pncB2 are two putative nicotinic acid phosphoribosyltransferases, playing a role in cof... more pncB1 and pncB2 are two putative nicotinic acid phosphoribosyltransferases, playing a role in cofactor salvage and drug resistance in Mycobacterium tuberculosis. Mutations have been reported in first- and second-line drug targets, causing resistance. However, pncB1 and pncB2 mutational data are not available, and neither of their mutation effects have been investigated in protein structures. The current study has been designed to investigate mutations and also their effects on pncB1 and pncB2 structures. A total of 287 whole-genome sequenced data of drug-resistant Mycobacterium tuberculosis isolates from Khyber Pakhtunkhwa of Pakistan were retrieved (BioSample PRJEB32684, ERR2510337-ERR2510445, ERR2510546-ERR2510645) from NCBI. The genomic data were analyzed for pncB1 and pncB2 mutations using PhyResSE. All the samples harbored numerous synonymous and non-synonymous mutations in pncB1 and pncB2 except one. Mutations Pro447Ser, Arg286Arg, Gly127Ser, and delTCAGGCCG1499213>1499220 ...
Therapeutic effects of anticancer medicines can be improved by targeting the specific receptors o... more Therapeutic effects of anticancer medicines can be improved by targeting the specific receptors on cancer cells. Folate receptor (FR) targeting with antibody (Ab) is an effective tool to deliver anticancer drugs to the cancer cell. In this research project, a novel formulation of targeting drug delivery was designed, and its anticancer effects were analyzed. Folic acid-conjugated magnetic nanoparticles (MNPs) were used for the purification of folate receptors through a novel magnetic affinity purification method. Antibodies against the folate receptors and methotrexate (MTX) were developed and characterized with enzyme-linked immunosorbent assay and Western blot. Targeting nanomedicines (MNP-MTX-FR Ab) were synthesized by engineering the MNP with methotrexate and anti-folate receptor antibody (anti-FR Ab). The cytotoxicity of nanomedicines on HeLa cells was analyzed by calculating the % age cell viability. A fluorescent study was performed with HeLa cells and tumor tissue sections t...
Pectinases hydrolyze pectin and make up 25% of global food processing enzyme sales. In this study... more Pectinases hydrolyze pectin and make up 25% of global food processing enzyme sales. In this study, we aimed to purify exo-polygalacturonase (Exo-PG) by using galacturonic acid conjugated magnetic nanoparticles (MNPs) and examined its application in juice purification. The submerged fermentation was carried out in the presence of apple pectin (1%) to promote production of exo-PG from Aspergillus flavus. Maximum exo-PG activity was observed after 4 days (30 °C and pH 5.0). A single protein band (66 kDa) of purified exo-PG was observed in SDS-PAGE. Purification of exo-PG enzyme was ∼ 10 fold with a yield of 29%. The enzyme retained 98% activity in the presence of 15 % glycerol at 4 °C. The purified exo-PG using MNPs yielded a 10-12% increase in juice production as compare to without treated fruit juice. To the best of our knowledge, this is the first report of affinity purification of exo-PG enzyme, using engineered magnetic nanoparticles.
Background Theranostic nanomedicines contain a nanovehicle that has fluorescent properties and ca... more Background Theranostic nanomedicines contain a nanovehicle that has fluorescent properties and can be used for diagnostic, therapeutic and prognostic purposes. The transferrin receptor expression is 1000-fold higher in rapidly growing cancer cells as compared to the normal cells and, therefore, can be used in targeted drug delivery systems. The objective of the present study was to design a novel targeted gold nanoparticle (GNPs)-based theranostic formulation for gastrointestinal (GI) tract-related cancers. The synthesized GNPs were conjugated to transferrin and doxorubicin both separately and collectively to check their cytotoxic properties. The in vitro cytotoxicity of nanocomposites was observed against colon cancer cell line HCT-116. The doxorubicin conjugated nanocomposites showed almost the same cytotoxicity, but more effect at later hours (h). The IC50 and IC100 were 50 µg/ml and 250 µg/ml, respectively, equivalent to the doxorubicin weight for GNP theranostic nanomedicine. R...
Genomic epidemiology of SARS-CoV-2 is imperative to explore the transmission, evolution, and also... more Genomic epidemiology of SARS-CoV-2 is imperative to explore the transmission, evolution, and also pathogenicity of viruses. The emergence of SARS-CoV-2 variants of concern posed a severe threat to the global public health efforts. To assess the potential consequence of these emerging variants on public health, continuous molecular epidemiology is of vital importance. The current study has been designed to investigate the major SARS-CoV-2 variants and emerging mutations in virus structural and non-structural proteins (NSP) during the fourth wave in September 2021 from the Punjab province of Pakistan. Twenty SARS-CoV-2 positive samples have been collected from major cities were subjected to next-generation sequencing. Among the 20 whole genomes (GenBank Accession SRR16294858-SRR16294877), 2 samples failed to be completely sequenced. These genome sequences harbored 207 non-synonymous mutations, among which 19 were unique to GISAID. The genome sequences were detected: Delta 21I, 21J var...
Antibodies are good escorts for a therapeutic and diagnostic tool against many diseases as well a... more Antibodies are good escorts for a therapeutic and diagnostic tool against many diseases as well as for targeting drug delivery and in immunohistochemistry. Cysteine-capped cadmium sulfide (quantum dot) nanoparticles have wide applications in immunohistochemistry due to their unique physical, chemical, and fluorescent properties. In this study, polyclonal antibodies against transferrin (as ligand), doxorubicin (anticancer drug), and H. pylori (cancer-causing agent) were developed in rabbits and purified. Purified antibodies were labeled with cadmium sulfide (CdS) quantum dot (QD) and used as a fluorescent-labeled marker for one-step identification in diagnosis and therapeutics. Cysteine-capped CdS QDs were synthesized in the presence of taurine (antioxidant) and characterized by FTIR, VSM, DLS, and TEM. CdS nanoparticles are monodisperse with a narrow size range below 7 nm and showed an increase of almost 30 nm after conjugation to IgG. The binding of QD-labeled antibodies was observ...
Exposure to carcinogenic chemicals, Helicobacter pylori infection, and high dietary salt are the ... more Exposure to carcinogenic chemicals, Helicobacter pylori infection, and high dietary salt are the risk factors associated with gastric cancer. Mice models of gastric cancer are key to understanding the cancer mechanism, to discerning the role played by different factors, and to determining therapeutic effects of different treatments. The goal has been to find targets which are only expressed with cancer so that they can be targeted specifically without harming normal cells. One such target could be the transferrin receptor, a glycoprotein receptor that is expressed many-folds on rapidly growing cells due to the greater demand of iron. In this study, gastric cancer was developed in mice (BALB/c) with human cancer-associated risk factors by feeding them with tumor-inducing concentration of methyl nitrosourea, dietary salt, and H. pylori along with normal feed and water. Three strategies were adopted to induce gastric cancer; (1) use of N-methyl-N-nitrosourea (MNU) with high dietary salt (NaCl), ( ) infection with H. pylori (isolated from human gastric tissue), and (3) use of MNU along with high concentration of NaCl after H. pylori infection. Mice were dissected after induction, and histological study of gastric tissue was done with Hematoxylin and Eosin staining. A diagnostic probe comprising transferrin conjugated with cadmium sulfide quantum dots was prepared and characterized. It was used to study the transferrin receptor overexpression in gastric tissue of cancer-induced mice relative to the normal mice. Mice of group 3 showed the highest rate of the cancer incidence ratio (96%) along with a high expression of transferrin receptors among the three groups. Histochemical studies showed that different types of gastric cancer depend upon the cancer-induction conditions. The mouse model of group 3 has the potential to be used in the future to study the therapeutic effects of cancer medicines, and overexpression of transferrin receptors could be identified through the designed probe to be used as diagnostics.
Structural proteins of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) are potential... more Structural proteins of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) are potential drug targets due to their role in the virus life cycle. The envelope (E) protein is one of the structural proteins; plays a critical role in virulency. However, the emergence of mutations oftenly leads to drug resistance and may also play a vital role in virus stabilization and evolution. In this study, we aimed to identify mutations in E proteins that affect the protein stability. About 0.3 million complete whole genome sequences were analyzed to screen mutations in E protein. All these mutations were subjected to stability prediction using the DynaMut server. The most common mutations that were detected at the C-terminal domain, Ser68Phe, Pro71Ser, and Leu73Phe, were examined through molecular dynamics (MD) simulations for a 100ns period. The sequence analysis shows the existence of 259 mutations in E protein. Interestingly, 16 of them were detected in the DFLV amino acid (aa) motif (aa72-aa75) that binds the host PALS1 protein. The results of root mean square deviation, fluctuations, radius of gyration, and free energy landscape show that Ser68Phe, Pro71Ser, and Leu73Phe are exhibiting a more stabilizing effect. However, a more comprehensive experimental study may be required to see the effect on virus pathogenicity. Potential antiviral drugs, and vaccines may be developed used after screening the genomic variations for better management of SARS-CoV-2 infections.
Natural bioactive compounds that trigger apoptosis in medicinal plants might be a promising new s... more Natural bioactive compounds that trigger apoptosis in medicinal plants might be a promising new source of anti-cancer medications. The present study aimed to assess the anti-cancer potential of the powerful therapeutic plant Chrysanthemum indicum. The whole plant extract was extracted in n-hexane and ethanol solvents to test anticancer and antioxidative capabilities on the cancer HepG2 cell line. The cells in the untreated group had not been exposed to any of the plant extracts. Ethanol and n-hexane extracts were administered to the treated cell lines. Cell viability was estimated by employing standard methodologies for the anticancer potential. Cell death and viability were measured by MTT, trypan blue, and crystal violet tests. The rate of apoptosis was measured in all groups using an Annexin-V ELISA. Furthermore, antioxidant enzyme activity studies were carried out to investigate the scavenging activities of nitric oxide, DPHH radicals, H2O2, and superoxide, scavenging power of t...
International Journal of Applied and Experimental Biology
Azadirachta indica constituents exhibit potential therapeutic role to control cancer progression.... more Azadirachta indica constituents exhibit potential therapeutic role to control cancer progression. The potential anti-cancerous activity of neem extracts reduces cell propagation, migration, inflammation, and invasion. The current review describes the importance and effectiveness of neem extracts in different solvents against various cancers. Studies performed in different geographic locations from 2001 to 2021 were searched in Google, PubMed, Google Scholar, and NCBI. The relevant information has been retrieved, including the type of extracts, solvents, year of publication, and type of cancers. A total of 125 relevant studies published in different journals have been screened, among which the most commonly used neem part was leaf (n = 79), followed by seed (n = 18), flower (n = 10), oil (n = 10), bark (n = 10), roots (n = 6), fruit (n = 6), gum (n = 5), limonoids (n = 5), nimbolide (n = 4), stem (n = 4), xylem (n = 1), and as herb (n = 1). Similarly, the most commonly investigated c...
pncB1 and pncB2 are two putative nicotinic acid phosphoribosyltransferases, playing a role in cof... more pncB1 and pncB2 are two putative nicotinic acid phosphoribosyltransferases, playing a role in cofactor salvage and drug resistance in Mycobacterium tuberculosis. Mutations have been reported in first- and second-line drug targets, causing resistance. However, pncB1 and pncB2 mutational data are not available, and neither of their mutation effects have been investigated in protein structures. The current study has been designed to investigate mutations and also their effects on pncB1 and pncB2 structures. A total of 287 whole-genome sequenced data of drug-resistant Mycobacterium tuberculosis isolates from Khyber Pakhtunkhwa of Pakistan were retrieved (BioSample PRJEB32684, ERR2510337-ERR2510445, ERR2510546-ERR2510645) from NCBI. The genomic data were analyzed for pncB1 and pncB2 mutations using PhyResSE. All the samples harbored numerous synonymous and non-synonymous mutations in pncB1 and pncB2 except one. Mutations Pro447Ser, Arg286Arg, Gly127Ser, and delTCAGGCCG1499213>1499220 ...
Therapeutic effects of anticancer medicines can be improved by targeting the specific receptors o... more Therapeutic effects of anticancer medicines can be improved by targeting the specific receptors on cancer cells. Folate receptor (FR) targeting with antibody (Ab) is an effective tool to deliver anticancer drugs to the cancer cell. In this research project, a novel formulation of targeting drug delivery was designed, and its anticancer effects were analyzed. Folic acid-conjugated magnetic nanoparticles (MNPs) were used for the purification of folate receptors through a novel magnetic affinity purification method. Antibodies against the folate receptors and methotrexate (MTX) were developed and characterized with enzyme-linked immunosorbent assay and Western blot. Targeting nanomedicines (MNP-MTX-FR Ab) were synthesized by engineering the MNP with methotrexate and anti-folate receptor antibody (anti-FR Ab). The cytotoxicity of nanomedicines on HeLa cells was analyzed by calculating the % age cell viability. A fluorescent study was performed with HeLa cells and tumor tissue sections t...
Pectinases hydrolyze pectin and make up 25% of global food processing enzyme sales. In this study... more Pectinases hydrolyze pectin and make up 25% of global food processing enzyme sales. In this study, we aimed to purify exo-polygalacturonase (Exo-PG) by using galacturonic acid conjugated magnetic nanoparticles (MNPs) and examined its application in juice purification. The submerged fermentation was carried out in the presence of apple pectin (1%) to promote production of exo-PG from Aspergillus flavus. Maximum exo-PG activity was observed after 4 days (30 °C and pH 5.0). A single protein band (66 kDa) of purified exo-PG was observed in SDS-PAGE. Purification of exo-PG enzyme was ∼ 10 fold with a yield of 29%. The enzyme retained 98% activity in the presence of 15 % glycerol at 4 °C. The purified exo-PG using MNPs yielded a 10-12% increase in juice production as compare to without treated fruit juice. To the best of our knowledge, this is the first report of affinity purification of exo-PG enzyme, using engineered magnetic nanoparticles.
Background Theranostic nanomedicines contain a nanovehicle that has fluorescent properties and ca... more Background Theranostic nanomedicines contain a nanovehicle that has fluorescent properties and can be used for diagnostic, therapeutic and prognostic purposes. The transferrin receptor expression is 1000-fold higher in rapidly growing cancer cells as compared to the normal cells and, therefore, can be used in targeted drug delivery systems. The objective of the present study was to design a novel targeted gold nanoparticle (GNPs)-based theranostic formulation for gastrointestinal (GI) tract-related cancers. The synthesized GNPs were conjugated to transferrin and doxorubicin both separately and collectively to check their cytotoxic properties. The in vitro cytotoxicity of nanocomposites was observed against colon cancer cell line HCT-116. The doxorubicin conjugated nanocomposites showed almost the same cytotoxicity, but more effect at later hours (h). The IC50 and IC100 were 50 µg/ml and 250 µg/ml, respectively, equivalent to the doxorubicin weight for GNP theranostic nanomedicine. R...
Genomic epidemiology of SARS-CoV-2 is imperative to explore the transmission, evolution, and also... more Genomic epidemiology of SARS-CoV-2 is imperative to explore the transmission, evolution, and also pathogenicity of viruses. The emergence of SARS-CoV-2 variants of concern posed a severe threat to the global public health efforts. To assess the potential consequence of these emerging variants on public health, continuous molecular epidemiology is of vital importance. The current study has been designed to investigate the major SARS-CoV-2 variants and emerging mutations in virus structural and non-structural proteins (NSP) during the fourth wave in September 2021 from the Punjab province of Pakistan. Twenty SARS-CoV-2 positive samples have been collected from major cities were subjected to next-generation sequencing. Among the 20 whole genomes (GenBank Accession SRR16294858-SRR16294877), 2 samples failed to be completely sequenced. These genome sequences harbored 207 non-synonymous mutations, among which 19 were unique to GISAID. The genome sequences were detected: Delta 21I, 21J var...
Antibodies are good escorts for a therapeutic and diagnostic tool against many diseases as well a... more Antibodies are good escorts for a therapeutic and diagnostic tool against many diseases as well as for targeting drug delivery and in immunohistochemistry. Cysteine-capped cadmium sulfide (quantum dot) nanoparticles have wide applications in immunohistochemistry due to their unique physical, chemical, and fluorescent properties. In this study, polyclonal antibodies against transferrin (as ligand), doxorubicin (anticancer drug), and H. pylori (cancer-causing agent) were developed in rabbits and purified. Purified antibodies were labeled with cadmium sulfide (CdS) quantum dot (QD) and used as a fluorescent-labeled marker for one-step identification in diagnosis and therapeutics. Cysteine-capped CdS QDs were synthesized in the presence of taurine (antioxidant) and characterized by FTIR, VSM, DLS, and TEM. CdS nanoparticles are monodisperse with a narrow size range below 7 nm and showed an increase of almost 30 nm after conjugation to IgG. The binding of QD-labeled antibodies was observ...
Exposure to carcinogenic chemicals, Helicobacter pylori infection, and high dietary salt are the ... more Exposure to carcinogenic chemicals, Helicobacter pylori infection, and high dietary salt are the risk factors associated with gastric cancer. Mice models of gastric cancer are key to understanding the cancer mechanism, to discerning the role played by different factors, and to determining therapeutic effects of different treatments. The goal has been to find targets which are only expressed with cancer so that they can be targeted specifically without harming normal cells. One such target could be the transferrin receptor, a glycoprotein receptor that is expressed many-folds on rapidly growing cells due to the greater demand of iron. In this study, gastric cancer was developed in mice (BALB/c) with human cancer-associated risk factors by feeding them with tumor-inducing concentration of methyl nitrosourea, dietary salt, and H. pylori along with normal feed and water. Three strategies were adopted to induce gastric cancer; (1) use of N-methyl-N-nitrosourea (MNU) with high dietary salt (NaCl), ( ) infection with H. pylori (isolated from human gastric tissue), and (3) use of MNU along with high concentration of NaCl after H. pylori infection. Mice were dissected after induction, and histological study of gastric tissue was done with Hematoxylin and Eosin staining. A diagnostic probe comprising transferrin conjugated with cadmium sulfide quantum dots was prepared and characterized. It was used to study the transferrin receptor overexpression in gastric tissue of cancer-induced mice relative to the normal mice. Mice of group 3 showed the highest rate of the cancer incidence ratio (96%) along with a high expression of transferrin receptors among the three groups. Histochemical studies showed that different types of gastric cancer depend upon the cancer-induction conditions. The mouse model of group 3 has the potential to be used in the future to study the therapeutic effects of cancer medicines, and overexpression of transferrin receptors could be identified through the designed probe to be used as diagnostics.
Structural proteins of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) are potential... more Structural proteins of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) are potential drug targets due to their role in the virus life cycle. The envelope (E) protein is one of the structural proteins; plays a critical role in virulency. However, the emergence of mutations oftenly leads to drug resistance and may also play a vital role in virus stabilization and evolution. In this study, we aimed to identify mutations in E proteins that affect the protein stability. About 0.3 million complete whole genome sequences were analyzed to screen mutations in E protein. All these mutations were subjected to stability prediction using the DynaMut server. The most common mutations that were detected at the C-terminal domain, Ser68Phe, Pro71Ser, and Leu73Phe, were examined through molecular dynamics (MD) simulations for a 100ns period. The sequence analysis shows the existence of 259 mutations in E protein. Interestingly, 16 of them were detected in the DFLV amino acid (aa) motif (aa72-aa75) that binds the host PALS1 protein. The results of root mean square deviation, fluctuations, radius of gyration, and free energy landscape show that Ser68Phe, Pro71Ser, and Leu73Phe are exhibiting a more stabilizing effect. However, a more comprehensive experimental study may be required to see the effect on virus pathogenicity. Potential antiviral drugs, and vaccines may be developed used after screening the genomic variations for better management of SARS-CoV-2 infections.
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