In silico Screening of Food and Drug Administration-approved Compounds against Trehalose 2-sulfotransferase (Rv0295c) in Mycobacterium tuberculosis: Insights from Molecular Docking and Dynamics Simulations.

BACKGROUND: Tuberculosis (TB) remains a prominent global health challenge, distinguished by substantial occurrences of infection and death. The upsurge of drug-resistant TB strains underscores the urgency to identify novel therapeutic targets and repurpose existing compounds. Rv0295c is a potentially druggable enzyme involved in cell wall biosynthesis and virulence. We evaluated the inhibitory activity of Food and Drug Administration (FDA)-approved compounds against Rv0295c of Mycobacterium tuberculosis, employing molecular docking, ADME evaluation, and dynamics simulations. METHODS: The study screened 1800 FDA-approved compounds and selected the top five compounds with the highest docking scores. Following this, we subjected the initially screened ligands to ADME analysis based on their dock scores. In addition, the compound exhibited the highest binding affinity chosen for molecular dynamics (MD) simulation to investigate the dynamic behavior of the ligand-receptor complex. RESULTS: Dihydroergotamine (CHEMBL1732) exhibited the highest binding affinity (-12.8 kcal/mol) for Rv0295c within this set of compounds. We evaluated the stability and binding modes of the complex over extended simulation trajectories. CONCLUSION: Our in silico analysis demonstrates that FDA-approved drugs can serve as potential Rv0295c inhibitors through repurposing. The combination of molecular docking and MD simulation offers a comprehensive understanding of the interactions between ligands and the protein target, providing valuable guidance for further experimental validation. Identifying Rv0295c inhibitors may contribute to new anti-TB drugs.

Differential proteome analysis of rat plasma after diisopropyl fluorophosphate (DFP) intoxication, a surrogate of nerve agent sarin.

Diisopropyl fluorophosphate (DFP), a surrogate of nerve agent sarin, is an organophosphorus (OP) compound which inhibits neuronal enzyme acetylcholinesterase (AChE). Exposure of this compound leads to a wide range of toxic symptoms and survivors may exhibit long term neurotoxicity related to cognitive and memory defects. Due to ease of availability and similar mechanism of action to other highly toxic nerve agent, DFP is widely used as model compound to trace changes associated with nerve agent exposures. Proximal fluids are widely used for the elucidation of biomarkers for exposure to toxic substances and to study the mechanism of toxicity. Using a rat model of OP intoxication, the present study was carried out to elucidate proteomic changes in plasma associated with DFP intoxication. Rats were exposed to a single dose (0.5 LD50) of DFP and their plasma proteome was studied, one day post exposure by two dimensional gel electrophoresis - mass spectrometry (2DE-MS). Some of the milestone changes were validated by Western blot analysis. A total 15 proteins showed significant fold changes in expression with respect to control after 1 day of DFP intoxication. Most of the proteins showing changes in expression at initial stages were related to immunogenic function, acute phase response, blood coagulation, and stress response. Experiments reported here demonstrate that 0.5 LD50 DFP intoxication leads to AChE inhibition, modulation of immunogenic function, and generation of stress at an early stage. Although, some proteins and their putative functional ramifications indicated similarity with those observed in our previous plasma proteome study, neurodegenerative changes were not observed in plasma of 0.5 LD50 DFP treated animals.

Expression profile of CXCL12 chemokine during M. tuberculosis infection with different therapeutic interventions in guinea pig.

Mycobacterium indicus pranii (MIP) already established as an immune-modulator in mycobacterial infections generates immune response by acting on CXC chemokines. In the present study, the immunomodulatory effect of MIP in conjunction with chemotherapy against M.tb infection was evaluated by colony forming units (CFUs) following aerosol infection to guinea pig and by measuring CXCL12 chemokine expression using q-PCR and in situ RT-PCR. Different experimental groups included, infection (Rv), immunoprophylaxis (RvMw), chemotherapy (RvCh) and combination of immunoprophylaxis+chemotherapy (RvChMw) group and normal healthy (NH) group. In the combination of immunoprophylaxis+chemotherapy (RvChMw) group, the CFU counts reduced significantly (p<0.001) at 4th week of infection as compared to other treated groups (RvMw and RvCh group). The expression of CXCL12 was recorded in all the treated groups of animals. The study demonstrated suppressed expression of CXCL 12 in both immunoprophylaxis as well as chemotherapy groups (6th and 8th week) that become elevated in immunoprophylaxis plus chemotherapy group (10th week), at which time point no CFUs were detected in RvCh and RvChMw group. The findings indicate that the expression of CXCL12 is associated with good response to anti - tubercular treatment. Thus, prior immunization with MIP appears to show good immunomodulatory effect to release CXCL12 chemokine during infection and also correlates with enhanced effect to chemotherapy.

From the Cover: Proteome Profile of Different Rat Brain Regions After Sarin Intoxication.

Sarin is an organophosphorus (OP) chemical warfare agent which irreversibly inhibits acetylcholinesterase. Acute toxicity after sarin exposure is because of hyper activation of the nicotinic and muscarinic receptor. Survivors of sarin exposure often develop long-term neuropathology referred as OP ester-induced chronic neurotoxicity. However, the exact mechanism of chronic neurotoxicity is yet unknown. We studied proteomic changes in rat brain regions after 0.5 LD50 dose of sarin and investigated some milestone changes associated with long-term CNS injury. We used two-dimensional gel electrophoresis/mass spectrometry approach to identify early proteomic changes and traced expression of selected proteins for longer time points. This study shows changes in chaperone function, endoplasmic reticulum stress, and defect in cytoskeleton functions at earlier stages. Predictive interaction analysis demonstrated putative role of Parkinson's disease-related proteins after sarin exposure. Our results clearly indicated neurodegenerative changes which started after 2.5 h and showed prominence after 3-month postexposure. The study also unmasks changes in proteins related to movement and cognitive function. The markers for astrocytosis (GFAP) and neurodegenerative changes (alpha-synuclein and amyloid precursor protein) exhibited altered expression in brain. This is the first proteomic study among survivors of sarin exposure in animal model. Some of the early changes, including those involved in neurodegeneration, movement, and cognitive function, defects in chaperone function and cytoskeleton, were shown to persist for a longer period. The study provides a preliminary framework for further validation of major mechanisms of sarin toxicity is suggested here and opens new avenues for elucidation of therapeutic intervention.

Botanical and Chemical Fingerprinting of Medicinal Roots of Justicia gendarussa Burm f.

BACKGROUND: Justicia gendarussa Burm f. of family Acanthaceae is medicinally important herb used in the treatment of inflammatory disorders, asthma, hepatic injuries, pathogenic infection and also shows antiproliferative activity against various cancer cell lines. MATERIALS AND METHODS: Pharmacognostical evaluation (macro-microscopy, physicochemical analysis and preliminary phytochemical analysis), high-performance thin layer chromatography (HPTLC) fingerprinting and chemical profiling by gas chromatography-mass spectrometry (GCMS) of dried roots of J. gendarussa were done according to quality standard procedures. RESULTS: Microscopic analysis revealed the compact arrangement of cells in cork region and thin-walled cortex beneath epidermis. Parenchymatous cells with xylem vessel were observed in the roots of J. gendarussa. Physicochemical studies revealed loss on drying (10.474%), total ash (2.990%), acid-insoluble ash (0.099%), water-soluble ash (1.528%), alcohol-soluble extractive value (0.564%) and water-soluble extractive value (4.11%) of the raw drug. Preliminary phytochemical analysis of ethanolic extract of J. gendarussa showed the presence of alkaloid, steroid, flavonoid, phenol, carbohydrate, saponin and quinone. Rf, color of the spots and densitometric scan were recorded by HPTLC fingerprinting using toluene: ethyl acetate: formic acid (5.0:4.0:1.0). On photodocumentation, six spots were visualized under 254 nm, nine spots under 360 nm and six spots at 620 nm. Identification of components in ethanolic extract of J. gendarussa was done by GC-MS. GC-MS results in the presence of oleic acid, 9,12-octadecadienoic acid, 6,9,12-octadecatrienoic acid and estra-1, 3,5 (10)-trein-17-ß-ol in ethanolic extract of J. gendarussa. CONCLUSION: These specific identities will be useful in identification and authentication of the raw drug in dried form. SUMMARY: Transverse section and powder of dried roots of Justicia gendarussa were examined microscopically. Microscopic observations showed the presence of well-developed cork and cortex. Presence of xylem vessels and parenchymatic rays were observed in transverse section. Parenchymatous cell and sclereids with vessel elements were found in powder microscopyPhysicochemical studies revealed loss on drying (10.474%), total ash (2.990%), acid-insoluble ash (0.099%), water-soluble ash (1.528%), alcohol-soluble extract (0.564%) and water-soluble extract (4.11%)Preliminary phytochemical analysis of ethanolic extract of J. gendarussa showed the presence of alkaloid, steroid, flavonoid, phenol, carbohydrate, saponin and quinoneHigh-performance thin layer chromatography fingerprinting showed different peaks at different wavelengthChemical profiling of medicinal roots of J. gendarussa by gas chromatography. mass spectrometry revealed the presence of oleic acid, 9,12.octadecadienoic acid, 6,9,12-octadecatrienoic acid and estra-1,3,5 (10)-trein-17-ß-ol as bioactive compound. Abbreviations Used: TLC: Thin layer chromatography; HPTLC: High performance thin layer chromatography; GCMS: Gas chromatography-mass specrtometry; QSIMP: Quality standard of indian medicinal plant; LOD: Loss on drying; TA: Total ash; AIA: Acid insoluble ash; WSA: Water soluble ash; ASE: Alcohol soluble extractive; WSE: Water soluble extractive.

Effect of Trigonella foenum graecum L on the Activities of Antioxidant Enzyme and Their Expression in Tissues of Alloxan-Induced Diabetic Rats.

Diabetes is a life-threatening metabolic disorder. This study was undertaken to evaluate the antihyperglycemic and antioxidative potential of seed powder of Trigonella foenum-graecum L in alloxan (55 mg/kg) induced diabetic rats. The results obtained showed that extensive oxidative stress is generated in tissues of diabetic rats as evidenced by increased production of hydrogen peroxide, increased accumulation of malondialdehyde (MDA) and 4-hydroxynonanal (4HNE) and decreased activities of superoxide dismutase (SOD), glutathione peroxidase (GPx), and catalase (CAT) in tissues of diabetic rats. It was observed that the transcription of genes of SOD, GPx, and CAT was also significantly decreased when compared with control. Treatment of Trigonella for 15 days to diabetic rats showed hypoglycemic effect and improved the altered levels of H2O2, MDA, and 4HNE, the activities of SOD, GPx, and CAT as well as transcription of these genes in the liver and the brain of diabetic rats.

Organophosphate pesticides-induced changes in the redox status of rat tissues and protective effects of antioxidant vitamins.

Organophosphates (OPs) pesticides are among the most toxic synthetic chemicals purposefully added in the environment. The common use of OP insecticides in public health and agriculture results in an environmental pollution and a number of acute and chronic poisoning events. Present study was aimed to evaluate the potential of monocrotophos and quinalphos to effect the redox status and glutathione (GSH) homeostasis in rat tissues and find out whether antioxidant vitamins have some protection on the pesticide-induced alterations. The results showed that these pesticides alone or in combination, caused decrease in the levels of GSH and the corresponding increase in the levels of GSSG, decreasing the GSH/GSSG ratio. The results also showed that NADPH/NADP(+) and NADH/NAD(+) ratios were decreased in the liver and brain of rats on exposure with mococrotophos, quinalphos, and their mixture. These pesticides, alone or in combination, caused alterations in the activities of GSH reductase and glucose-6-phosphate dehydrogenase in the rat tissues. However, the expression of the GSH recycling enzymes did not show significant alterations as compared to control. From the results, it can be concluded that these pesticides generate oxidative stress but their effects were not synergistic when given together and prior feeding of antioxidant vitamins tend to reduce the toxicities of these pesticides.

Chloroquine efficacy studies confirm drug susceptibility of Plasmodium vivax in Chennai, India.

BACKGROUND: Assessing the Plasmodium vivax burden in India is complicated by the potential threat of an emerging chloroquine (CQ) resistant parasite population from neighbouring countries in Southeast Asia. Chennai, the capital of Tamil Nadu and an urban setting for P. vivax in southern India, was selected as a sentinel site for investigating CQ efficacy and sensitivity in vivax malaria. METHODS: CQ efficacy was evaluated with a 28-day in vivo therapeutic study, while CQ sensitivity was measured with an in vitro drug susceptibility assay. In both studies, isolates also underwent molecular genotyping to investigate correlations between parasite diversity and drug susceptibility to CQ. Molecular genotyping included sequencing a 604 base pair (bp) fragment of the P. vivax multidrug resistant gene-1 (Pvmdr1) for single nucleotide polymorphisms (SNPs) and also the amplification of eight microsatellite (MS) loci located across the genome on eight different chromosomes. RESULTS: In the 28-day in vivo study (N=125), all subjects were aparasitaemic by Day 14. Passive case surveillance continuing beyond Day 28 in 22 subjects exposed 17 recurrent infections, which ranged from 44 to 148 days post-enrollment. Pvmdr1 sequencing of these recurrent infections revealed that 93.3% had identical mutant haplotypes (958M/Y976/1076L) to their baseline Day 0 infection. MS genotyping further revealed that nine infection pairs were related with ≥ 75% haplotype similarity (same allele at six or more loci). To test the impact of this mutation on CQ efficacy, an in vitro drug assay (N=68) was performed. No correlation between IC50 values and the percentage of ring-stage parasites prior to culture was observed (r(sadj): -0.00063, p = 0.3307) and the distribution of alleles among the Pvmdr1 SNPs and MS haplotypes showed no significant associations with IC50 values. CONCLUSIONS: Plasmodium vivax was found to be susceptible to CQ drug treatment in both the in vivo therapeutic drug study and the in vitro drug assay. Though the mutant 1076 L of Pvmdr1 was found in a majority of isolates tested, this single mutation did not associate with CQ resistance. MS haplotypes revealed strong heterogeneity in this population, indicating a low probability of reinfection with highly related haplotypes.

A sturgeon-derived bioactive compound beneficially modulates nuclear receptors controlling metabolic functions in patients with metabolic syndrome.

The aim of the present study was to test the possible effects of a novel sturgeon-derived compound  (LD-1227) on inflammatory markers related to metabolic nuclear receptors in patients with metabolic syndrome. The study population consisted of 76 patients with metabolic syndrome and 30 healthy subjects who were maintained to their current treatments and randomly supplemented: A) LD-1227 (n=38) or B) placebo (n=38) as compared to C) healthy controls (n=30). LD-1227 or placebo (water-soluble starch) were given daily at breakfast and dinner for three months. Levels of hs-CRP, IL-6, TNF-α, leptin and adiponectin/ resistin index were assayed at the entry, 1 month and 3 months afterwards. At the end of the study period, as compared to B group, LD-1227-treated patients showed a significant improvement of all parameters tested, irrespective of the presence of diabetes. In particular, levels of adiponectin and adiponectin/ resistin index significantly increased following LD-1227 administration. Although the metabolic syndrome remains a multifaceted condition requiring a complex approach, LD-1227 could be a potential safe therapeutic tool to be integrated into a wider treatment and preventive medicine schedule strategy.

Evaluation of DNA damage and cytotoxicity induced by three commonly used organophosphate pesticides individually and in mixture, in rat tissues.

Organophosphate pesticides are among the most widely used synthetic chemicals for controlling a wide variety of pests. Chlorpyrifos (CPF), methyl parathion (MPT), and malathion (MLT) are among the most extensively used organophosphate (OP) pesticides. The main target of action of OP compounds is the central and peripheral nervous system, although it has also been postulated that these compounds in both acute and chronic intoxication, disturb the redox processes and thus induce oxidative stress. The excessive generation of reactive oxygen species (ROS) causes damage to all vital macromolecules including lipids, proteins, and DNA. This study was aimed to investigate the genotoxicity and cytotoxicity of CPF, MPT, and MLT when given singly or in combination. The DNA damage was measured by alkaline single-cell gel electrophoresis or comet assay and expressed as DNA damage index. The results showed that both acute and chronic exposure with CPF, MPT, and MLT, caused significantly marked DNA damage in rat tissues namely, liver, brain, kidney, and spleen, when measured 24 hour posttreatment. It was also observed that MPT caused highest level of DNA damage and brain was maximally affected by these OP compounds. When these pesticides were given in mixture, the damage was not the sum of damage caused by individual pesticide, confirming that these pesticides do not potentiate the toxicity of each other. When the DNA damage was measured 48 and 72 hour posttreatment, the damage was partially repaired. Pesticide exposure also caused histopathological changes in rat tissues.

Genotoxic Potential of Reactive Oxygen Species (Ros), Lipid Peroxidation and DNA Repair Enzymes (Fpg and Endo III) in Alloxan Injected Diabetic Rats.

Diabetes is a chronic metabolic syndrome due to insulin deficiency and is associated with increased oxidative stress in vivo. Oxidative stress including, increased production of reactive oxygen species (ROS) in vivo, can lead to cellular biomolecule damage. Such damage has been suggested to contribute to the pathogenesis of diabetes mellitus. Genotoxicity induced by ROS in diabetic rats, was estimated by measuring DNA single strand breaks and double strand breaks by standard comet assay/ single cell gel electrophoresis (SCGE). To find out whether DNA lesions were caused due to oxidative stress, combination of bacterial DNA repair enzymes which convert base damage to breaks are used. Formamidoaminopyrimidine glycosylase (Fpg) and Endonuclese (Endo III)] recognize oxidized purines and oxidized pyrimidines, respectively, were used in modified comet assay. Significant increase in DNA strand breaks in terms of DNA damage index were observed in diabetic rat lymphocytes in modified comet assay. The involvement of oxidative stress was also examined by estimation of thiobarbituric acid reactive substances (TBARS) in diabetic rats. The levels of TBARS, reactive oxygen species (ROS) namely, hydrogen peroxide, superoxide and nitrate/nitrite anion were also increased in diabetic rats that further shows the involvement of oxidative stress in ROS induced DNA damage. The results of the present study show genotoxic potential of ROS in diabetic rats.

Evaluation of micronuclei induction capacity and mutagenicity of organochlorine and organophosphate pesticides.

The genotoxic and mutagenic effects of two commonly used organochlorine pesticides, lindane (LND) and endosulfan (ENS), and two commonly used organophosphate pesticides, chlorpyrifos (CPF) and monocrotophos (MCP) were assessed using in vivo mouse bone marrow micronucleus test and in vitro Ames Salmonella/ microsome mutagenicity test. The results showed that these pesticides alone or in combination, induced significantly high frequency of micronuclei (MN) formation that increased with concentration of pesticides. All these four pesticides produced significant increase in the frequencies of micronucleated-polychromatic erythrocytes (MN-PCE) and decrease infrequencies of PCE in dose-dependent manner. The results indicate the suppression of proliferative activity of the bone marrow and increase in the extent of cell death. ENS and MCP showed mutagenic potential in Salmonella/ microsome assay. ENS induced mutagenic and nontoxic response only in TA98 tester strain of S.typhimurium at the dose of 500 µg/plate and in the absence of metabolic activation. MCP showed weak mutagenic and nontoxic effect only in TA100 tester strain at the dose of 5000 µg/plate in both assays, with or without metabolic activation when compared with negative control. MCP was toxic in TA98 tester strain at the dose of 5000 µg/plate in absence of metabolic activation while reduction in toxicity was seen on addition of S9 mixture. The study clearly showed the genotoxic potential of all these four pesticides and mutagenic response of endosulfan and monocrotophos.

Inferring the evolutionary history of Indian Plasmodium vivax from population genetic analyses of multilocus nuclear DNA fragments.

The human malaria parasite Plasmodium vivax is globally widespread, causing high malaria morbidity. As P. vivax is highly endemic to India, and previous reports indicate genetic homogeneity in population samples, we tested the hypothesis of no genetic structuring in Indian P. vivax. Further, based on the reports of increasing incidence of Plasmodium falciparum infection in comparison with P. vivax in recent years in India, it was important to understand whether reduction in population size has resulted in decrease in P. vivax infection rate in India. For this, we utilized recently developed putatively neutral markers from chromosome 13 of P. vivax to score single nucleotide polymorphisms in 126 P. vivax isolates collected from 10 different places in India. The overall results indicated that Indian P. vivax bears high nucleotide diversity within population samples but moderate amount of genetic differentiation between population samples. STRUCTURE analysis grouped 10 population samples into three clusters based on the proportion of the genetic ancestries in each population. However, the pattern of clustering does not correlate with sampling locations in India. Furthermore, analyses of past demographic events indicated reduction in population size in majority of population samples, but when isolates from all the 10 samples were considered as a single population, the data fit to the demographic equilibrium model. All these observations clearly indicate that Indian P. vivax presents complex evolutionary history but possesses several features of being a part of ancestral distribution range of this species.

Redox imbalance in rat tissues exposed with organophosphate pesticides and therapeutic potential of antioxidant vitamins.

Organophosphate pesticides are among the most widely used synthetic chemicals for controlling domestic and agricultural pests. Present study was aimed to evaluate the potential of chlorpyrifos, parathion and malathion, to disturb glutathione homeostasis in rat tissues and to find out whether the pre-feeding of antioxidant vitamins has some ameliorating effect on the pesticide-induced alterations. The results showed that these pesticides, alone or in combination, caused decrease in the levels of GSH and the corresponding increase in the levels of GSSG, decreasing the GSH/GSSG ratio. The results also showed NADPH/NADP(+) and NADH/NAD(+) ratios were also decreased in the rat tissues on pesticide exposure. These pesticides, alone or in combination, caused increase in the activities of glutathione reductase and glucose-6-phosphate dehydrogenase in all the rat tissues studied. The findings show that these pesticides generate oxidative stress and prior feeding of mixture of antioxidant vitamins tend to reduce the toxicities of these pesticides.

The effect of lovastatin on oxidative stress and antioxidant enzymes in hydrogen peroxide intoxicated rat.

Oxidative stress has been linked to the development of many diseases and hastens the progression of cardiovascular diseases. Since lovastatin is used worldwide as a cholesterol lowering drug, the present study was undertaken to evaluate the antioxidant property of lovastatin against H(2)O(2) induced oxidative stress in rats. Four study groups of rats of four animals each were treated with DMSO (control), H(2)O(2) (OS), lovastatin (L) and H(2)O(2) + lovastatin (OSL). On the 15th day the animals were sacrificed, and the liver and heart tissues were analyzed for oxidative stress biomarkers and anti-oxidant enzymes. Results of the OSL-group showed a reduction in thiobarbituric acid reactive substances in liver (42.7%) and heart tissue (8%) compared with the control group. An increase was observed in the activity of the antioxidant enzymes, catalase (34.6% in liver and 33.3% in heart) and glutathione peroxidase (50.5% in liver and 34.7% in heart). A commensurate increase in the activity of G6PDH was observed indicating an enhanced requirement of NADPH. The ratio GSH:GSSG in liver (1.05) and heart (0.84) was satisfactorily regulated compared to the control group (1.01 in liver and 0.93 in heart). These results suggest that lovastatin possesses antioxidant activity and reduces oxidative stress.

Regional alterations of JNK3 and CaMKIIα subunit expression in the rat brain after soman poisoning.

Calcium/calmodulin-dependent protein kinase II (CaMKII) and c-Jun N-terminal kinases (JNKs) exert numerous and diverse functions in the brain. However, their role in nerve agent poisoning is poorly understood. In the present study, rats were exposed to soman (80 µg/kg) subcutaneously to study the changes in the protein levels of calcium/calmodulin-dependent protein kinase II alpha subunit (CaMKIIα) and JNK3 and activities of acetylcholinestarase (AChE) and CaMKII in the rat brain. Western blot analysis revealed that significant changes were found in both the protein kinases expression. Immunoreactivity levels of neural specific JNK3 isoform increased from 2.5 hours to 30 days after soman exposure in cerebral cortex, hippocampus, striatum and thalamus regions and decreased in the case of cerebellum. CaMKIIα expression levels were also increased from 2.5 hours to 30 days after soman exposure in cerebral cortex, hippocampus, thalamus and down regulated in cerebellum. AChE activity remained inhibited in plasma and brain up to 3 days post exposure. CaMKII activity was increased in cerebrum and decreased in cerebellum. Results suggest that altered expression of both the protein kinases play a role in nerve agent-induced long-term neurotoxic effects.

Identification of differentially displayed proteins in cerebrospinal fluid of Parkinson's disease patients: a proteomic approach.

BACKGROUND: Clinical proteomics has been widely used to identify differentially displayed proteins in blood and cerebrospinal fluid (CSF) to understand the molecular and cellular events leading to Parkinson's disease (PD). The close connection between CSF and the brain offers reliable and reproducible way to assess the majority of changes in the brain proteome profile directly into CSF throughout the course of neurodegeneration. METHODS: We identified the differentially displayed proteins in CSF of PD patients as compared with controls using two-dimensional polyacrylamide gel electrophoresis (2-D PAGE) and mass spectrometry. RESULTS: Comparative 2-D PAGE electrophoretograms of CSF of PD patients with case controls and/or neurological controls revealed significant differential display of six protein spots. The differentially displayed proteins were identified as serum albumin precursor, serum albumin chain-A, hemoglobin beta fragment, mutant globin, proline rich repeat 14 (PRR 14) and serum transferrin N-terminal lobe. Although the level of hemoglobin beta fragment and mutant globin was attenuated, serum albumin precursor, serum albumin chain-A, PRR 14 and serum transferrin N-terminal lobe were augmented in PD patients as compared with case controls. The level of serum albumin chain-A, PRR 14 and serum transferrin N-terminal lobe was not significantly altered when compared with neurological controls. CONCLUSIONS: The results obtained thus suggest that differential display of CSF serum albumin precursor, serum albumin chain-A, PRR 14 and serum transferrin N-terminal lobe could be associated with neuronal dysfunction and hemoglobin/globin with the onset/progression of PD in humans.

Chlorpyrifos-induced DNA damage in rat liver and brain.

Chlorpyrifos (O,O'-diethyl-O-3,5,6-trichloro-2-pyridyl phosphorothionate, CPF) is a broad spectrum organophosphate pesticide used to control a variety of pests. The present study was undertaken to test the in vivo genotoxic potential of CPF in rats, using the single cell gel electrophoresis (or comet) assay. The rats were administered 50 mg and 100 mg CPF/kg body weight daily for 1, 2, and 3 days as well as 1.12 mg and 2.24 mg CPF/kg body weight for 90 days. The level of DNA damage was estimated by scoring 100 cells per animal, dividing into five types: types 0, I, II, III, and IV. The results clearly indicate that exposure to CPF, acutely or chronically, caused a dose-dependent increase in DNA damage in the liver and brain of rats. From the present study, it can be concluded that CPF exhibits genotoxic potential in vivo.

Functional restoration using basic fibroblast growth factor (bFGF) infusion in Kainic acid induced cognitive dysfunction in rat: neurobehavioural and neurochemical studies.

Neurogenesis occurs in dentate gyrus of adult hippocampus under the influence of various mitogenic factors. Growth factors besides instigating the proliferation of neuronal progenitor cells (NPCs) in dentate gyrus, also supports their differentiation to cholinergic neurons. In the present study, an attempt has been made to investigate the neurotrophic effect of bFGF in Kainic acid (KA) induced cognitive dysfunction in rats. Stereotaxic lesioning using (KA) was performed in hippocampal CA3 region of rat's brain. Four-weeks post lesioning rats were assessed for impairment in learning and memory using Y maze followed by bFGF infusion in dentate gyrus region. The recovery was evaluated after bFGF infusion using neurochemical, neurobehavioural and immunohistochemical approaches and compared with lesioned group. Significant impairment in learning and memory (P < 0.01) observed in lesioned animals, four weeks post lesioning exhibited significant restoration (P < 0.001) following bFGF infusion twice at one and four week post lesion. The bFGF infused animals exhibited recovery in hippocampus cholinergic (76%)/ dopaminergic (46%) receptor binding and enhanced Choline acetyltransferase (ChAT) immunoreactivity in CA3 region. The results suggest restorative potential of bFGF in cognitive dysfunctions, possibly due to mitogenic effect on dentate gyrus neurogenic area leading to generation and migration of newer cholinergic neurons.

Effect of chlorpyrifos on thiobarbituric acid reactive substances, scavenging enzymes and glutathione in rat tissues.

The present study showed that exposure of chlorpyrifos, O,O'-diethyl-O-3,5,6-trichloro-2-pyridyl phosphorothionate (CPF), a widely used pesticide in rats caused significant inhibition of acetylcholinesterase (AChE) activity in different tissues viz., liver, kidney and spleen. CPF exposure also generated oxidative stress in the body, as evidenced by increase in thiobarbituric acid reactive substances (TBARS), decrease in the levels of superoxide scavenging enzymes viz., superoxide dismutase (SOD), catalase (CAT) and glutathione peroxidase (GPx) in liver, kidney and spleen at all doses. Malondialdehyde levels were increased by 14%, 31% and 76% in liver, 11%, 31% and 64% in kidney and 32%, 75% and 99.9% in spleen when 50 mg, 100 mg and 200 mg/kg body wt. CPF was administered for three days. SOD and CAT activities were decreased in liver, kidney and spleen, while GPx activity showed slight increase in kidney at 50 mg and 100 mg dose, and decreased on further increase in dose of CPF. Liver and spleen showed dose-dependent decrease in GPx activity. The levels of reduced glutathione (GSH) was decreased, while oxidized glutathione (GSSG) was increased, thus a marked fall in GSH/GSSG ratio was observed in all tissues. A maximum decrease of 83% was observed in liver, followed by kidney and spleen, which showed 78% and 57% decrease, respectively in group given 200 mg/kg CPF. The levels of glucose-6-phosphate dehydrogenase (G6PDH) and glutathione reductase (GR) were also decreased in liver and kidney, while spleen showed increase at lower doses, but decrease at high dose of CPF. The data provide evidence for induction of oxidative stress on CPF exposure.