DESCRIPTION The research on Human glutathione s-transferase a1-1 with naturally occurring compoun... more DESCRIPTION The research on Human glutathione s-transferase a1-1 with naturally occurring compounds may provide valuable insights and their role as chemotherapeutic agents. Inhibition study of selenium-glutathione peroxidase with mercaptans using in silco approach provides new insight into the mechanism of glutathione peroxidase. I have identified the ellagic acid analogues (CID 56611575, CID 46229200 and CID 10003463) as a novel potent CK2 inhibitor. The biological activities of these compounds in terms of IC50 were calculated based on generated QSAR model.
Mycobacterium tuberculosis (M.tb.) enoyl-acyl carrier protein (ACP) reductase (InhA) is validated... more Mycobacterium tuberculosis (M.tb.) enoyl-acyl carrier protein (ACP) reductase (InhA) is validated as a useful target for tuberculosis therapy and is considered an attractive enzyme to drug discovery. This study aimed to identify the novel inhibitor of the InhA enzyme, a potential target of M.tb. involved in the type II fatty acid biosynthesis pathway that controls mycobacterial cell envelope synthesis. We compiled 80 active compounds from Ruta graveolens and citrus plants belonging to the Rutaceae family for pharmacokinetics and molecular docking analyses. The chemical structures of the 80 phytochemicals and the 3D structure of the target protein were retrieved from the PubChem database and RCSB Protein Data Bank, respectively. The evaluation of druglikeness was performed based on Lipinski’s Rule of Five, while the computed phytochemical properties and molecular descriptors were used to predict the ADMET of the compounds. Amongst these, 11 pharmacokinetically-screened compounds were...
Objectives: To investigate whether inhalable microparticles containing two anti-tuberculosis agen... more Objectives: To investigate whether inhalable microparticles containing two anti-tuberculosis agents, isoniazid and rifampicin, evoke host-defence strategies in macrophages in addition to targeting the incorporated drugs. Methods: Microparticles were prepared by spray-drying a homogeneous solution of drugs and poly (lactic acid) (PLA; apparent viscosity 1.1 cP). Four parts PLA and three parts rifampicin were dissolved in dichloromethane. One part isoniazid was dissolved in methanol. The two solutions were mixed in the ratio 22: 3 at which none of the solutes precipitated. These were administered as ‘nose-only ’ inha-lations to mice or exposed to cultured J774 mouse macrophages. Targeting to lung macrophages was investigated by transmission electron microscopy. Reactive oxygen species (ROS) were estimated by a cytochrome c assay and flow cytometry. Reactive nitrogen intermediates (RNI) were assayed using Griess reagent. Cytokines in culture supernatants were estimated by ELISA. Result...
Modern-day agriculture is evolving from the traditional production of raw food products to advanc... more Modern-day agriculture is evolving from the traditional production of raw food products to advances in novel food engineering technologies that ensure purity and functionality including health-specific products. Plants are the ultimate source of food and nutrition. Nutraceuticals are the functional foods which can provide health and medicinal benefits or can be used for prevention and treatment of various diseases along with providing basic nutrition. Nutraceuticals can be purified food nutrients, dietary supplements, herbs, cereals, milk, soups, or herbal products to genetically engineered foods enriched with vitamins and essential minerals. The components may also include phytochemicals, probiotics, vitamins, antioxidants, and essential minerals that are derived from plant and/or microbial sources. Recently, nanoparticle pharmaceutical drug delivery systems came into picture. These nanocarriers can also be used to enhance the potential of nano-formulated nutraceuticals. Presently,...
AIMS Macrophage is known to readily engulf any particulate material they encounter, including inv... more AIMS Macrophage is known to readily engulf any particulate material they encounter, including invading microbes and nano- or micro-particles. While recent studies show that some microparticles (MP) are immunogenic even without drug-cargo, the mechanism underlying this phenomenon is yet unclear. Phagocytosis induces NADPH oxidase-2 (NOX-2) mediated ROS generation that is reported to regulate antibacterial autophagy. We therefore, investigated the role of NOX-2 derived ROS in phagosomal maturation and autophagy induction in response to phagocytic uptake of two kinds of polymeric biodegradable and biocompatible microparticles: yeast-derived β-glucan particles (YDGP) and poly-(D, L-Lactic Acid) microparticles (PMP). MAIN METHODS J774A.1 macrophage was exposed to polymeric particles and the immune responses: ROS, phagosomal maturation and autophagy induction, were examined by assays including NBT, DCFH-DA, NADPH-Oxidase activity, Lysotracker and Acridine Orange. Further, the LC3 and NOX-...
Leptospira is a zoonotic pathogen causing significant morbidity and mortality both in animals and... more Leptospira is a zoonotic pathogen causing significant morbidity and mortality both in animals and humans. Although several surface proteins have been identified as vaccine candidate, they failed to induce sterilizing immunity and cross protection against different serovars. Thus, identification of highly immunogenic antigens that are conserved among pathogenic serovars would be first step towards development of universal vaccine for Leptospirosis. Here we used reverse vaccinology pipeline to screen core genome of pathogenic Leptospira spp.in order to identify suitable vaccine candidates. Based on properties like sub cellular localization, adhesin, homology to human proteins, antigenicity and allergenicity, 18 antigenic proteins were identified and were further investigated for immunological properties. Based on immunogenicity, Protegenicity, Antigenicity, B-cell and promiscuous T-cell epitopes, 6 Potential Vaccine Candidates (PVCs) were finally selected which covered most of the aff...
International Journal of Health Sciences and Research, 2014
Background and Aims: Antituberculosis drug-induced hepatotoxicity (anti-TB-DIH) is a growing prob... more Background and Aims: Antituberculosis drug-induced hepatotoxicity (anti-TB-DIH) is a growing problem of serious concern in many countries around the world. Drug metabolizing enzymes have been proposed to play an influential role in the pathogenesis of anti-TB-DIH. Therefore, the main aim of this study was to investigate the possible association of GSTM1, GSTT1 and CYP2E1 gene polymorphisms with anti-TB-DIH. Methods: A prospective case-control study was conducted in 244 North Indian TB patients receiving anti-TB treatment. These patients were prospectively followed up both clinically and biochemically, before and during the treatment period, for the diagnosis of anti-TB-DIH. The frequencies of GSTM1, GSTT1 and CYP2E1 gene polymorphisms were determined from genomic DNA, utilizing multiplex polymerase chain reaction and restriction fragment length polymorphism methods respectively. Logistic regression analysis was used to determine the association between gene polymorphisms and anti-TB...
We investigated the preclinical efficacy and safety/tolerability of biodegradable polymeric parti... more We investigated the preclinical efficacy and safety/tolerability of biodegradable polymeric particles containing isoniazid (INH) and rifabutin (RFB) dry powder for inhalation (DPI) as an adjunct to oral first-line therapy. Mice and guinea pigs infected with Mycobacterium tuberculosis H37Rv (Mtb) were treated with ∼80 and ∼300 μg of the DPI, respectively, for 3-4 weeks starting 3, 10, and 30 days post-infection. Adjunct combination therapy eliminated culturable Mtb from the lungs and spleens of all but one of 52 animals that received the DPI. Relapse-free cure was not achieved in one mouse that received DPI + oral, human-equivalent doses (HED) of four drugs used in the Directly Observed Treatment, Short Course (DOTS), starting 30 days post-infection. Oral doses (20 mg/Kg/day, each) of INH + RFB reduced Mtb burden from ∼106 to ∼103 colony-forming units. Combining half the oral dose with DPI prevented relapse of infection four weeks after stopping the treatment. The DPI was safe in rod...
Macrophage responses to infection with Mycobacterium tuberculosis (MTB) and treatment with solubl... more Macrophage responses to infection with Mycobacterium tuberculosis (MTB) and treatment with soluble isoniazid (INH) plus rifabutin (RFB) versus microparticles containing equivalent amounts of drugs were compared. It was investigated whether macrophages driven to alternative activation upon infection with MTB could be rescued to display the classical activation phenotype. It was established that microparticles sustain high levels of drugs in cytosol of macrophages for longer period as compared to soluble drugs. Microparticles co-localized with intracellular bacteria, and induced a variety of innate bactericidal responses, including induction of free radicals, alteration of mitochondrial membrane potential and apoptosis. The data strongly suggest that additional benefit may be derived from the nature of the drug delivery system, which fulfils Koch's dictum 'stimulate the phagocyte' for curing tuberculosis.
β-glucan particles (GP) are polymeric carbohydrates, mainly found as components of cell wall fung... more β-glucan particles (GP) are polymeric carbohydrates, mainly found as components of cell wall fungi, yeast, bacteria and also in cereals such as barley and oat, and have been recently shown to have application in macrophage-targeted drug delivery. The aim of this study was to prepare and characterize GP containing a large payload of Rifabutin (RB), an anti-tuberculosis drug effective against MDR-TB at lower MIC than Rifampicin. GP were prepared from yeast cells by acidic and alkaline extraction were either spray dried or lyophilized, prior to RB loading and alginate sealing. The FTIR and (13)C-NMR spectra of the GP confirmed a β-(1→3) linked glucan structure, with a triple-helical conformation. The spray dried GP exhibited better characteristics in terms of uniformity, size range (2.9 to 6.1 µm) and more than 75 % particles were below 3.5 μm. The RP-HPLC analysis of spray dried GP revealed drug entrapment and drug loading up to 81.46 ± 4.9 % and ~40.5 ± 1.9 %, respectively, as compar...
Beta-glucans are carbohydrates (glucose polymers) found in the cell walls of fungi, yeast, algae,... more Beta-glucans are carbohydrates (glucose polymers) found in the cell walls of fungi, yeast, algae, lichens, and plants such as oats and barley. Beta-glucans bind to glucan receptor on phagocytic cells and modify these cells to become "immunologically active" by generating a variety of innate immune responses. Particulate beta-glucan has been specifically shown to engage dectin-1 receptor, which leads to the recruitment and activation of nicotinamide adenine dinucleotide phosphate oxidase-2 (NOX-2) and release of antimicrobial reactive oxygen species (ROS). Here, we report that yeast-derived beta-glucan particles (YDGP) of a specific size are easily phagocytosed by macrophages and induce the production of ROS. Furthermore, the present work also demonstrates that phagosomal maturation, appearance of acidic vesicular compartments (AVO), and light chain protein-3 (LC3) accumulation within murine macrophages, occur at early and delayed time points after the phagocytic uptake of ...
DESCRIPTION The research on Human glutathione s-transferase a1-1 with naturally occurring compoun... more DESCRIPTION The research on Human glutathione s-transferase a1-1 with naturally occurring compounds may provide valuable insights and their role as chemotherapeutic agents. Inhibition study of selenium-glutathione peroxidase with mercaptans using in silco approach provides new insight into the mechanism of glutathione peroxidase. I have identified the ellagic acid analogues (CID 56611575, CID 46229200 and CID 10003463) as a novel potent CK2 inhibitor. The biological activities of these compounds in terms of IC50 were calculated based on generated QSAR model.
Background and Aims: Antituberculosis drug-induced hepatotoxicity (anti-TB-DIH) is a growing prob... more Background and Aims: Antituberculosis drug-induced hepatotoxicity (anti-TB-DIH) is a growing problem of serious concern in many countries around the world. Drug metabolizing enzymes have been proposed to play an influential role in the pathogenesis of anti-TB-DIH. Therefore, the main aim of this study was to investigate the possible association of GSTM1, GSTT1 and CYP2E1 gene polymorphisms with anti-TB-DIH. Methods: A prospective case-control study was conducted in 244 North Indian TB patients receiving anti-TB treatment. These patients were prospectively followed up both clinically and biochemically, before and during the treatment period, for the diagnosis of anti-TB-DIH. The frequencies of GSTM1, GSTT1 and CYP2E1 gene polymorphisms were determined from genomic DNA, utilizing multiplex polymerase chain reaction and restriction fragment length polymorphism methods respectively. Logistic regression analysis was used to determine the association between gene polymorphisms and anti-TB...
DESCRIPTION The research on Human glutathione s-transferase a1-1 with naturally occurring compoun... more DESCRIPTION The research on Human glutathione s-transferase a1-1 with naturally occurring compounds may provide valuable insights and their role as chemotherapeutic agents. Inhibition study of selenium-glutathione peroxidase with mercaptans using in silco approach provides new insight into the mechanism of glutathione peroxidase. I have identified the ellagic acid analogues (CID 56611575, CID 46229200 and CID 10003463) as a novel potent CK2 inhibitor. The biological activities of these compounds in terms of IC50 were calculated based on generated QSAR model.
Mycobacterium tuberculosis (M.tb.) enoyl-acyl carrier protein (ACP) reductase (InhA) is validated... more Mycobacterium tuberculosis (M.tb.) enoyl-acyl carrier protein (ACP) reductase (InhA) is validated as a useful target for tuberculosis therapy and is considered an attractive enzyme to drug discovery. This study aimed to identify the novel inhibitor of the InhA enzyme, a potential target of M.tb. involved in the type II fatty acid biosynthesis pathway that controls mycobacterial cell envelope synthesis. We compiled 80 active compounds from Ruta graveolens and citrus plants belonging to the Rutaceae family for pharmacokinetics and molecular docking analyses. The chemical structures of the 80 phytochemicals and the 3D structure of the target protein were retrieved from the PubChem database and RCSB Protein Data Bank, respectively. The evaluation of druglikeness was performed based on Lipinski’s Rule of Five, while the computed phytochemical properties and molecular descriptors were used to predict the ADMET of the compounds. Amongst these, 11 pharmacokinetically-screened compounds were...
Objectives: To investigate whether inhalable microparticles containing two anti-tuberculosis agen... more Objectives: To investigate whether inhalable microparticles containing two anti-tuberculosis agents, isoniazid and rifampicin, evoke host-defence strategies in macrophages in addition to targeting the incorporated drugs. Methods: Microparticles were prepared by spray-drying a homogeneous solution of drugs and poly (lactic acid) (PLA; apparent viscosity 1.1 cP). Four parts PLA and three parts rifampicin were dissolved in dichloromethane. One part isoniazid was dissolved in methanol. The two solutions were mixed in the ratio 22: 3 at which none of the solutes precipitated. These were administered as ‘nose-only ’ inha-lations to mice or exposed to cultured J774 mouse macrophages. Targeting to lung macrophages was investigated by transmission electron microscopy. Reactive oxygen species (ROS) were estimated by a cytochrome c assay and flow cytometry. Reactive nitrogen intermediates (RNI) were assayed using Griess reagent. Cytokines in culture supernatants were estimated by ELISA. Result...
Modern-day agriculture is evolving from the traditional production of raw food products to advanc... more Modern-day agriculture is evolving from the traditional production of raw food products to advances in novel food engineering technologies that ensure purity and functionality including health-specific products. Plants are the ultimate source of food and nutrition. Nutraceuticals are the functional foods which can provide health and medicinal benefits or can be used for prevention and treatment of various diseases along with providing basic nutrition. Nutraceuticals can be purified food nutrients, dietary supplements, herbs, cereals, milk, soups, or herbal products to genetically engineered foods enriched with vitamins and essential minerals. The components may also include phytochemicals, probiotics, vitamins, antioxidants, and essential minerals that are derived from plant and/or microbial sources. Recently, nanoparticle pharmaceutical drug delivery systems came into picture. These nanocarriers can also be used to enhance the potential of nano-formulated nutraceuticals. Presently,...
AIMS Macrophage is known to readily engulf any particulate material they encounter, including inv... more AIMS Macrophage is known to readily engulf any particulate material they encounter, including invading microbes and nano- or micro-particles. While recent studies show that some microparticles (MP) are immunogenic even without drug-cargo, the mechanism underlying this phenomenon is yet unclear. Phagocytosis induces NADPH oxidase-2 (NOX-2) mediated ROS generation that is reported to regulate antibacterial autophagy. We therefore, investigated the role of NOX-2 derived ROS in phagosomal maturation and autophagy induction in response to phagocytic uptake of two kinds of polymeric biodegradable and biocompatible microparticles: yeast-derived β-glucan particles (YDGP) and poly-(D, L-Lactic Acid) microparticles (PMP). MAIN METHODS J774A.1 macrophage was exposed to polymeric particles and the immune responses: ROS, phagosomal maturation and autophagy induction, were examined by assays including NBT, DCFH-DA, NADPH-Oxidase activity, Lysotracker and Acridine Orange. Further, the LC3 and NOX-...
Leptospira is a zoonotic pathogen causing significant morbidity and mortality both in animals and... more Leptospira is a zoonotic pathogen causing significant morbidity and mortality both in animals and humans. Although several surface proteins have been identified as vaccine candidate, they failed to induce sterilizing immunity and cross protection against different serovars. Thus, identification of highly immunogenic antigens that are conserved among pathogenic serovars would be first step towards development of universal vaccine for Leptospirosis. Here we used reverse vaccinology pipeline to screen core genome of pathogenic Leptospira spp.in order to identify suitable vaccine candidates. Based on properties like sub cellular localization, adhesin, homology to human proteins, antigenicity and allergenicity, 18 antigenic proteins were identified and were further investigated for immunological properties. Based on immunogenicity, Protegenicity, Antigenicity, B-cell and promiscuous T-cell epitopes, 6 Potential Vaccine Candidates (PVCs) were finally selected which covered most of the aff...
International Journal of Health Sciences and Research, 2014
Background and Aims: Antituberculosis drug-induced hepatotoxicity (anti-TB-DIH) is a growing prob... more Background and Aims: Antituberculosis drug-induced hepatotoxicity (anti-TB-DIH) is a growing problem of serious concern in many countries around the world. Drug metabolizing enzymes have been proposed to play an influential role in the pathogenesis of anti-TB-DIH. Therefore, the main aim of this study was to investigate the possible association of GSTM1, GSTT1 and CYP2E1 gene polymorphisms with anti-TB-DIH. Methods: A prospective case-control study was conducted in 244 North Indian TB patients receiving anti-TB treatment. These patients were prospectively followed up both clinically and biochemically, before and during the treatment period, for the diagnosis of anti-TB-DIH. The frequencies of GSTM1, GSTT1 and CYP2E1 gene polymorphisms were determined from genomic DNA, utilizing multiplex polymerase chain reaction and restriction fragment length polymorphism methods respectively. Logistic regression analysis was used to determine the association between gene polymorphisms and anti-TB...
We investigated the preclinical efficacy and safety/tolerability of biodegradable polymeric parti... more We investigated the preclinical efficacy and safety/tolerability of biodegradable polymeric particles containing isoniazid (INH) and rifabutin (RFB) dry powder for inhalation (DPI) as an adjunct to oral first-line therapy. Mice and guinea pigs infected with Mycobacterium tuberculosis H37Rv (Mtb) were treated with ∼80 and ∼300 μg of the DPI, respectively, for 3-4 weeks starting 3, 10, and 30 days post-infection. Adjunct combination therapy eliminated culturable Mtb from the lungs and spleens of all but one of 52 animals that received the DPI. Relapse-free cure was not achieved in one mouse that received DPI + oral, human-equivalent doses (HED) of four drugs used in the Directly Observed Treatment, Short Course (DOTS), starting 30 days post-infection. Oral doses (20 mg/Kg/day, each) of INH + RFB reduced Mtb burden from ∼106 to ∼103 colony-forming units. Combining half the oral dose with DPI prevented relapse of infection four weeks after stopping the treatment. The DPI was safe in rod...
Macrophage responses to infection with Mycobacterium tuberculosis (MTB) and treatment with solubl... more Macrophage responses to infection with Mycobacterium tuberculosis (MTB) and treatment with soluble isoniazid (INH) plus rifabutin (RFB) versus microparticles containing equivalent amounts of drugs were compared. It was investigated whether macrophages driven to alternative activation upon infection with MTB could be rescued to display the classical activation phenotype. It was established that microparticles sustain high levels of drugs in cytosol of macrophages for longer period as compared to soluble drugs. Microparticles co-localized with intracellular bacteria, and induced a variety of innate bactericidal responses, including induction of free radicals, alteration of mitochondrial membrane potential and apoptosis. The data strongly suggest that additional benefit may be derived from the nature of the drug delivery system, which fulfils Koch's dictum 'stimulate the phagocyte' for curing tuberculosis.
β-glucan particles (GP) are polymeric carbohydrates, mainly found as components of cell wall fung... more β-glucan particles (GP) are polymeric carbohydrates, mainly found as components of cell wall fungi, yeast, bacteria and also in cereals such as barley and oat, and have been recently shown to have application in macrophage-targeted drug delivery. The aim of this study was to prepare and characterize GP containing a large payload of Rifabutin (RB), an anti-tuberculosis drug effective against MDR-TB at lower MIC than Rifampicin. GP were prepared from yeast cells by acidic and alkaline extraction were either spray dried or lyophilized, prior to RB loading and alginate sealing. The FTIR and (13)C-NMR spectra of the GP confirmed a β-(1→3) linked glucan structure, with a triple-helical conformation. The spray dried GP exhibited better characteristics in terms of uniformity, size range (2.9 to 6.1 µm) and more than 75 % particles were below 3.5 μm. The RP-HPLC analysis of spray dried GP revealed drug entrapment and drug loading up to 81.46 ± 4.9 % and ~40.5 ± 1.9 %, respectively, as compar...
Beta-glucans are carbohydrates (glucose polymers) found in the cell walls of fungi, yeast, algae,... more Beta-glucans are carbohydrates (glucose polymers) found in the cell walls of fungi, yeast, algae, lichens, and plants such as oats and barley. Beta-glucans bind to glucan receptor on phagocytic cells and modify these cells to become "immunologically active" by generating a variety of innate immune responses. Particulate beta-glucan has been specifically shown to engage dectin-1 receptor, which leads to the recruitment and activation of nicotinamide adenine dinucleotide phosphate oxidase-2 (NOX-2) and release of antimicrobial reactive oxygen species (ROS). Here, we report that yeast-derived beta-glucan particles (YDGP) of a specific size are easily phagocytosed by macrophages and induce the production of ROS. Furthermore, the present work also demonstrates that phagosomal maturation, appearance of acidic vesicular compartments (AVO), and light chain protein-3 (LC3) accumulation within murine macrophages, occur at early and delayed time points after the phagocytic uptake of ...
DESCRIPTION The research on Human glutathione s-transferase a1-1 with naturally occurring compoun... more DESCRIPTION The research on Human glutathione s-transferase a1-1 with naturally occurring compounds may provide valuable insights and their role as chemotherapeutic agents. Inhibition study of selenium-glutathione peroxidase with mercaptans using in silco approach provides new insight into the mechanism of glutathione peroxidase. I have identified the ellagic acid analogues (CID 56611575, CID 46229200 and CID 10003463) as a novel potent CK2 inhibitor. The biological activities of these compounds in terms of IC50 were calculated based on generated QSAR model.
Background and Aims: Antituberculosis drug-induced hepatotoxicity (anti-TB-DIH) is a growing prob... more Background and Aims: Antituberculosis drug-induced hepatotoxicity (anti-TB-DIH) is a growing problem of serious concern in many countries around the world. Drug metabolizing enzymes have been proposed to play an influential role in the pathogenesis of anti-TB-DIH. Therefore, the main aim of this study was to investigate the possible association of GSTM1, GSTT1 and CYP2E1 gene polymorphisms with anti-TB-DIH. Methods: A prospective case-control study was conducted in 244 North Indian TB patients receiving anti-TB treatment. These patients were prospectively followed up both clinically and biochemically, before and during the treatment period, for the diagnosis of anti-TB-DIH. The frequencies of GSTM1, GSTT1 and CYP2E1 gene polymorphisms were determined from genomic DNA, utilizing multiplex polymerase chain reaction and restriction fragment length polymorphism methods respectively. Logistic regression analysis was used to determine the association between gene polymorphisms and anti-TB...
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Papers by Rolee Sharma