Author Report for: Sharma SNo contact information in database for Sharma S
Mudgal R, Sharma S, Singh S, Ravichandiran V Ref: Front Neurol, 14:1130575, 2023 : PubMed ESTHER: Mudgal_2023_Front.Neurol_14_1130575 PubMedSearch: Mudgal 2023 Front.Neurol 14 1130575 PubMedID: 37153653 Abstract Imidacloprid (IMI) is not only a neurotoxic agricultural pesticide but also a possible food contaminant. The aims of this study were to (1) explore the relationship between recurrent IMI administration and neuronal toxicity in mice and (2) evaluate the potential neuroprotective effect of ascorbic acid (AA), a substance with significant free radical scavenger and having property to block the inflammatory pathways. Mice were categorized as naive controls (administered vehicles for 28 days); the IMI-treatment animal group (administered po 45-mg/kg body weight of IMI per day for 28 days); and the IMI + AA treatment animal group (administered the same IMI dose + 200 mg/kg of AA orally for 28 days). On day 28, memory losses were assessed using the Y-maze and novel target identification behavioral tests. Mice were sacrificed 24 h after the final IMI treatments, as well as hippocampus tissues, were utilized to determine histological assessments, oxidative stress biomarkers, and Heme oxygenase-1 (HO-1) and nuclear factor erythroid 2-related factor 2 (Nrf2) gene expression levels. The findings demonstrated that IMI-treated mice had substantial impairment of spatial and non-spatial memory functions, as well as reduced antioxidant enzyme and acetylcholinesterase activity. The AA neuroprotective action was achieved through the suppression of the HO-1 expression as well as the stimulation of Nrf2 expression in hippocampal tissues. In summary, recurrent IMI exposure causes oxidative stress and neurotoxicity in mice, and the administration of AA significantly reduces the IMI toxicity possibly by the activation of the HO-1/Nrf2 pathway. Title: Structural and Functional Characterization of Mycobacterium tuberculosis Homoserine Transacetylase Sharma S, Jayasinghe YP, Mishra NK, Orimoloye MO, Wong TY, Dalluge JJ, Ronning DR, Aldrich CC Ref: ACS Infect Dis, :, 2023 : PubMed ESTHER: Sharma_2023_ACS.Infect.Dis__ PubMedSearch: Sharma 2023 ACS.Infect.Dis PubMedID: 36753622 Gene_locus related to this paper: myctu-metx Abstract Mycobacterium tuberculosis (Mtb) lacking functional homoserine transacetylase (HTA) is compromised in methionine biosynthesis, protein synthesis, and in the activity of multiple essential S-adenosyl-l-methionine-dependent enzymes. Additionally, deficient mutants are further disarmed by the toxic accumulation of lysine due to a redirection of the metabolic flux toward the lysine biosynthetic pathway. Studies with deletion mutants and crystallographic studies of the apoenzyme have, respectively, validated Mtb HTA as an essential enzyme and revealed a ligandable binding site. Seeking a mechanistic characterization of this enzyme, we report crucial structural details and comprehensive functional characterization of Mtb HTA. Crystallographic and mass spectral observation of the acetylated HTA intermediate and initial velocity studies were consistent with a ping-pong kinetic mechanism. Wild-type HTA and its site-directed mutants were kinetically characterized with a panel of natural and alternative substrates to understand substrate specificity and identify critical residues for catalysis. Titration experiments using fluorescence quenching showed that both substratesacetyl-CoA and l-homoserineengage in a strong and weak binding interaction with HTA. Additionally, substrate inhibition by acetyl-CoA and product inhibition by CoA and O-acetyl-l-homoserine were proposed to form the basis of a feedback regulation mechanism. By furnishing key mechanistic and structural information, these studies provide a foundation for structure-based design efforts around this attractive Mtb target. Title: Screening of Multitarget-Directed Natural Compounds as Drug Candidates for Alzheimer's Disease Using In Silico Techniques: Their Extraction and In Vitro Validation Srivastava S, Sharma S, Deep S, Khare SK Ref: ACS Omega, 8:38118, 2023 : PubMed ESTHER: Srivastava_2023_ACS.Omega_8_38118 PubMedSearch: Srivastava 2023 ACS.Omega 8 38118 PubMedID: 37867692 Abstract Alzheimer's disease (AD) is a neurodegenerative disorder that impairs neurocognitive function. Acetylcholinesterase (AChE) and beta-site APP cleaving enzyme 1 (BACE1) are the two main proteins implicated in AD. Indeed, the major available commercial drugs (donepezil, rivastigmine, and galantamine) against Alzheimer's are AChE inhibitors. However, none of these drugs are known to reverse or reduce the pathophysiological condition of the disease since there are multiple contributing factors to AD. Therefore, there is a need to develop a multitarget-directed ligand approach for its treatment. In the present study, plant bioactive compounds were screened for their AChE and BACE1 inhibition potential by conducting molecular docking studies. Considering their docking score and pharmacokinetic properties, limonin, peimisine, serratanine B, and withanolide A were selected as the lead compounds. Molecular dynamics simulations of these protein-ligand complexes confirmed the conformational and energetically stabilized enzyme-inhibitor complexes. The inhibition potential of the lead compounds was validated by in vitro enzyme assay. Withanolide A inhibited AChE (IC(50) value of 107 microM) and showed mixed-type inhibition. At this concentration, it inhibited BACE1 activity by 57.10% and was stated as most effective. Both the compounds, as well as their crude extracts, were found to have no cytotoxic effect on the SH-SY5Y cell line. Title: Repurposing of digoxin in pain and inflammation: An evidence-based study Patel S, Gururani R, Jain S, Tripathi N, Paliwal S, Sharma S Ref: Drug Dev Res, :, 2022 : PubMed ESTHER: Patel_2022_Drug.Dev.Res__ PubMedSearch: Patel 2022 Drug.Dev.Res PubMedID: 35315525 Abstract In recent years, the drug repositioning strategy has gained considerable attention in the drug discovery process that involves establishing new therapeutic uses of already known drugs. In line with this, we have identified digoxin a cardiac glycoside, as a potent inhibitor of soluble epoxide hydrolase (sEH) enzyme employing in silico high throughput screening protocols and further confirmed using in vitro cell-free sEH inhibitory assay and in vivo preclinical studies in rodents for its repurposing in hyperalgesia, inflammation, and related disorders. Oral administration of digoxin at dose 0.2 mg/kg significantly reduced (p < .0001) the allodynia in mice induced by using hot plate (3.6 +/- 1.9) and tail-flick test (7.58 +/- 0.9). In addition, digoxin at a dose of 0.2 mg/kg showed marked reduction (94%, p < .0001) in acetic acid-induced abdominal contraction in rats. Further, digoxin also demonstrated antipyretic activity (37.04 +/- 0.2, p < .0001) and showed notable reduction (0.60 +/- 0.06) in carrageenan-induced paw edema in rats. Also, the histopathological evaluation revealed that digoxin treatment attenuated the edema, neutrophil infiltration, and alveolar septal thickening in lung tissue. These findings are novel and highlight the newer insights towards repurposing digoxin as a new lead in the treatment of hyperalgesia, inflammation, and related disorders. Title: Repositioning of tubocurarine as analgesic and anti-inflammatory agent: Exploring beyond myorelaxant activity Patel S, Shukla J, Jain S, Paliwal V, Tripathi N, Paliwal S, Sharma S Ref: Biochemical Pharmacology, 205:115248, 2022 : PubMed ESTHER: Patel_2022_Biochem.Pharmacol_205_115248 PubMedSearch: Patel 2022 Biochem.Pharmacol 205 115248 PubMedID: 36113566 Abstract BACKGROUND AND PURPOSE: Tubocurarine (d-TC), a non-depolarizing competitive blocker of nicotinic acetylcholine receptors is extensively utilized for the relaxation of skeletal muscles. Drug repositioning is a forthright approach to reduce the cost and speed up drug development process. Herein, we have attempted to evaluate the analgesic and anti-inflammatory activity of d-TC for its possible repurposing in pain and inflammation-related issues. EXPERIMENTAL APPROACH: We examined the soluble epoxide hydrolase inhibitory (sEHI) activity of d-TC employing in silico high throughput screening protocols, in vitro cell-free sEH inhibitory assay, and in in vivo rodent models for its repositioning in pain and inflammation-related disorders. KEY RESULTS: In molecular docking study, d-TC displayed impressive hydrogen bonding interactions within the cavity of sEH enzyme with good docking score. d-TC also exhibited notable sEH inhibitory activity (IC(50) 3.72 nm) at the in vitro assay. Oral absorption capability of d-TC (0.1 and 0.2 mg/mL) was determined using an in vitro everted intestinal sac model employing rat ileum tissue that revealed significant oral absorption of d-TC. Besides, in vivo studies revealed that oral administration of d-TC (0.1 and 0.2 mg/kg) in rodents significantly attenuated hyperalgesia (cold plate test, tail immersion test and formalin test) and inflammation (estimation of rectal temperature, acetic acid induced pleurisy test and cotton pellet-induced granuloma test) induced in robust preclinical models. Conclusion and implications These findings are novel and warrant immediate efforts to reposition d-TC as a new therapeutic candidate in the management of hyperalgesia, inflammation, and associated conditions. Title: Mechano-chemical and biological energetics of immobilized enzymes onto functionalized polymers and their applications Sharma T, Xia C, Sharma A, Raizada P, Singh P, Sharma S, Sharma P, Kumar S, Lam S, Nadda AK Ref: Bioengineered, 13:10518, 2022 : PubMed ESTHER: Sharma_2022_Bioengineered_13_10518 PubMedSearch: Sharma 2022 Bioengineered 13 10518 PubMedID: 35443858 Abstract Enzymes of commercial importance, such as lipase, amylase, laccase, phytase, carbonic anhydrase, pectinase, maltase, glucose oxidase etc., show multifunctional features and have been extensively used in several fields including fine chemicals, environmental, pharmaceutical, cosmetics, energy, food industry, agriculture and nutraceutical etc. The deployment of biocatalyst in harsh industrial conditions has some limitations, such as poor stability. These drawbacks can be overcome by immobilizing the enzyme in order to boost the operational stability, catalytic activity along with facilitating the reuse of biocatalyst. Nowadays, functionalized polymers and composites have gained increasing attention as an innovative material for immobilizing the industrially important enzyme. The different types of polymeric materials and composites are pectin, agarose, cellulose, nanofibers, gelatin, and chitosan. The functionalization of these materials enhances the loading capacity of the enzyme by providing more functional groups to the polymeric material and hence enhancing the enzyme immobilization efficiency. However, appropriate coordination among the functionalized polymeric materials and enzymes of interest plays an important role in producing emerging biocatalysts with improved properties. The optimal coordination at a biological, physical, and chemical level is requisite to develop an industrial biocatalyst. Bio-catalysis has become vital aspect in pharmaceutical and chemical industries for synthesis of value-added chemicals. The present review describes the current advances in enzyme immobilization on functionalized polymers and composites. Furthermore, the applications of immobilized enzymes in various sectors including bioremediation, biosensor and biodiesel are also discussed. Title: Donepezil-Inspired Multitargeting Indanone Derivatives as Effective Anti-Alzheimer's Agents Singh JV, Thakur S, Kumar N, Singh H, Mithu VS, Bhagat K, Gulati HK, Sharma A, Sharma S, Bedi PMS Ref: ACS Chem Neurosci, :, 2022 : PubMed ESTHER: Singh_2022_ACS.Chem.Neurosci__ PubMedSearch: Singh 2022 ACS.Chem.Neurosci PubMedID: 35195392 Abstract In continuous efforts to develop anti-Alzheimer's agents, we rationally designed and synthesized a series of multitargeting molecules by incorporating the essential molecular features of the standard drug donepezil. Among the series, compound 4b showed multitargeting properties to act as an anti-Alzheimer's agent, which is better tolerable in vivo than donepezil. Acetylcholinesterase (AChE) inhibition data showed that compound 4b inhibits the enzyme with a half-maximal inhibitory concentration (IC(50)) value of 0.78 microM and also showed DNA protection, which was confirmed through the DNA nicking assay, suggesting the protective effect of 4b against oxidative DNA damage. Compound 4b also showed 53.04% inhibition against Abeta(1-42) aggregations, which was found comparable to that of the standard compound curcumin. Molecular dynamics simulations were performed to check the stability of compound 4b with the enzyme AChE, which showed that the enzyme-ligand complex is stable enough to block the hydrolysis of acetylcholine in the brain. Its higher LD(50) cutoff value (50 mg/kg) in comparison to donepezil (LD(50): 25 mg/kg) made it safer, suggesting that it can be used in further clinical experiments. To evaluate its anti-Alzheimer property, a mice model with melamine-induced cognitive dysfunction was used, and Morris water maze and Rotarod tests were performed. A significant improvement in memory was observed after the treatment with compound 4b and donepezil. The study postulated that the introduction of important structural features of donepezil (dimethoxyindanone moiety as ring-A) embarked with terminal aromatic ether (ring-B and ring-C) made 4b a multitargeting molecule that offers a way for developing alternative therapeutics in the future against Alzheimer's disease (AD). Title: Therapeutic Potential of Reserpine in Metabolic Syndrome: An Evidence Based Study Verma K, Paliwal S, Sharma S Ref: Pharmacol Res, :106531, 2022 : PubMed ESTHER: Verma_2022_Pharmacol.Res__106531 PubMedSearch: Verma 2022 Pharmacol.Res 106531 PubMedID: 36336214 Abstract Reserpine is as old as the scientific diagnosis of hypertension. For many years' clinicians have used it for the treatment of high blood pressure, but with the passage of time and introduction of new anti-hypertensive drugs, the usage of reserpine has gone down drastically most probably due to poorly understood mechanism of action and multiple misleading adverse effects precisely due to high dosing of reserpine. With an aim to elucidate the specific mechanism of action, we screened reserpine against various targets associated with regulation of blood pressure. Surprisingly reserpine showed remarkable inhibitory potential for soluble epoxide hydrolase an enzyme responsible for pathophysiology of not only hypertension but also hyperlipidemia, diabetes and inflammation collectively known as metabolic syndrome. The in-silico, in-vitro and in-vivo results showed that reserpine has the ability to treat metabolic syndrome effectively by inhibiting soluble epoxide hydrolase. Title: N-Acetylcysteine Ameliorates Neurotoxic Effects of Manganese Intoxication in Rats: A Biochemical and Behavioral Study Chopra D, Sharma S, Sharma N, Nehru B Ref: Neurochem Res, :, 2021 : PubMed ESTHER: Chopra_2021_Neurochem.Res__ PubMedSearch: Chopra 2021 Neurochem.Res PubMedID: 33950473 Abstract Clinical and experimental evidences reveal that excess exposure to manganese is neurotoxic and leads to cellular damage. However, the mechanism underlying manganese neurotoxicity remains poorly understood but oxidative stress has been implicated to be one of the key pathophysiological features related to it. The present study investigates the effects associated with manganese induced toxicity in rats and further to combat these alterations with a well-known antioxidant N-acetylcysteine which is being used in mitigating the damage by its radical scavenging activity. The study was designed to note the sequential changes along with the motor and memory dysfunction associated with biochemical and histo-pathological alterations following exposure and treatment for 2 weeks. The results so obtained showed decrease in the body weights, behavioral deficits with increased stress markers and also neuronal degeneration in histo-pathological examination after manganese intoxication in rats. To overcome the neurotoxic effects of manganese, N-acetylcysteine was used in the current study due to its pleiotropic potential in several pathological ailments. Taken together, N-acetylcysteine helped in ameliorating manganese induced neurotoxic effects by diminishing the behavioral deficits, normalizing acetylcholinesterase activity, and augmentation of redox status. Title: A combinatorial approach to screen structurally diverse acetylcholinesterase inhibitory plant secondary metabolites targeting Alzheimer's disease Choudhir G, Sharma S, Hariprasad P Ref: J Biomol Struct Dyn, :1, 2021 : PubMed ESTHER: Choudhir_2021_J.Biomol.Struct.Dyn__1 PubMedSearch: Choudhir 2021 J.Biomol.Struct.Dyn 1 PubMedID: 34351840 Abstract Alzheimer's disease (AD) is a form of Dementia known to diminish the brain's function by perturbating its structural and functional components. Though cholinesterase inhibitors are widely used to treat AD, they are limited by numbers and side effects. Hence, present study aims to identify structurally diverse Acetylcholinesterase (AChE) inhibitory plant secondary metabolites (PSM) by employing high throughput screening and computational studies. AChE inhibitory activity was performed using 390 crude extracts from 63 plant parts belongs to 58 plants. The lowest IC(50) value was recorded by acetone extract of Cyperus rotundus rhizome at 0.5 mg/ml, followed by methanol extract of Terminalia arjuna bark (0.95 mg/ml) and water extract Acacia catechu stem (0.95 mg/ml). A virtual library containing 487 PSM belongs to 18 plants found positive for AChE inhibition (IC(50)>=5 mg/ml) was prepared. Through ADMET analysis, 78 PSM fulfilling selected drug-likeness parameters were selected for further analysis. Molecular docking studies of selected PSM against AChE recorded a wide range of binding energy from -3.40 to -10.90 Kcal/mol. Further molecular dynamics simulation studies also recorded stabilized interactions of AChE-ligand complexes in the term of RMSD, RMSF, Rg, SASA, and hydrogen bond interaction. MMPBSA analysis revealed the binding energy of selected PSM ranging from -123.757 to -261.697 kJ/mol. Our study demonstrated the potential of 12 PSM (Sugiol, Margolone, 7-Hydroxy-3',4'-(Methylenedioxy) flavan, Beta-cyprone, Ethenone, Isomargolonone, Serpentine, Cryptolepine, Rotundone, Strictamin, Rotundenol and Nootkatone) as AChE inhibitors. Further in vitro and in vivo experimental evaluations with pure PSM could be beneficial for therapeutic uses. Communicated by Ramaswamy H. Sarma. Title: A clinical perspective of soluble epoxide hydrolase inhibitors in metabolic and related cardiovascular diseases Verma K, Jain S, Paliwal S, Sharma S Ref: Curr Molecular Pharmacology, :, 2021 : PubMed ESTHER: Verma_2021_Curr.Mol.Pharmacol__ PubMedSearch: Verma 2021 Curr.Mol.Pharmacol PubMedID: 34544352 Abstract Epoxide hydrolase (EH) is a crucial enzyme responsible for catabolism, detoxification, and regulation of signaling molecules in various organisms including human beings. In mammals, EHs are classified according to their DNA sequence, sub-cellular location, and activity into eight major classes: soluble EH (sEH), microsomal EH (mEH), leukotriene A4 hydrolase (LTA4H), cholesterol EH (ChEH), hepoxilin EH, paternally expressed gene 1 (peg1/MEST), EH3 and EH4. The sEH, an alpha/beta-hydrolase fold family enzyme is an emerging pharmacological target in multiple diseases namely, cardiovascular disease, neurodegenerative disease, chronic pain, fibrosis, diabetes, pulmonary diseases, and immunological disease. It exhibits prominent physiological effect that includes anti-inflammatory, anti-migratory and vasodilatory effects. Its efficacy has been documented in several kinds of clinical trials and observational studies. This review specifically highlights the development of soluble epoxide hydrolase inhibitors (sEHIs) in the clinical setting for the management of metabolic syndrome and related disorders such as cardiovascular effects, endothelial dysfunction, arterial disease, hypertension, diabetes, obesity, heart failure, and dyslipidemia. In addition, limitations and future aspects of sEHIs have also been highlighted which will help the investigators to bring the sEHI to the clinics. Title: Genetic polymorphisms in the mEH gene in relation to tobacco smoking: role in lung cancer susceptibility and survival in north Indian patients with lung cancer undergoing platinum-based chemotherapy Walia HK, Singh N, Sharma S Ref: Future Oncol, :, 2021 : PubMed ESTHER: Walia_2021_Future.Oncol__ PubMedSearch: Walia 2021 Future.Oncol PubMedID: 34672683 Abstract Aim: Epoxide hydrolase is involved in oxidative defenses and is responsible for the activation of carcinogens. The relationship between EPHX1 polymorphisms (Tyr(113)His and His(139)Arg) and overall survival (OS) and lung cancer (LC) risk was investigated. Methods: The study comprised 550 cases and 550 controls. Genotyping and statistical analysis were applied. Results: The variant genotypes of EPHX1 polymorphisms exhibited no association with LC risk. The Tyr(113)His polymorphism exhibited twofold increased odds of lymph node invasion (p = 0.04). The Tyr/His genotype is a risk factor for smokers. Subjects carrying the combined genotype for His(139)Arg showed better median survival time (MST) and the heterozygous genotype revealed better MST in the case of small-cell lung cancer (SCLC; 11.30 vs 6.73 months; log-rank test: p = 0.02). The heterozygous genotype (His139Arg) had longer MST in patients receiving cisplatin/carboplatin and irinotecan (11.30 vs 7.23; log-rank test: p = 0.007) Conclusion: The Tyr(113)His polymorphism is associated with LC risk in smokers and is a potential prognostic factor for OS in patients with SCLC after irinotecan. Title: Biomarkers for the toxicity of sublethal concentrations of triclosan to the early life stages of carps Dar OI, Sharma S, Singh K, Sharma A, Bhardwaj R, Kaur A Ref: Sci Rep, 10:17322, 2020 : PubMed ESTHER: Dar_2020_Sci.Rep_10_17322 PubMedSearch: Dar 2020 Sci.Rep 10 17322 PubMedID: 33057045 Abstract Accumulation, contents of protein, non-enzymatic antioxidant glutathione (GSH and GSSG), lipid peroxidation product (melondialdehyde-MDA) and organic acids (fumarate, succinate, malate and citrate), and activities of neurological (acetylcholinesterase-AChE), detoxification (glutathione S-transferase-GST) and metabolic (lactate dehydrogenase-LDH, aspartate transaminase-AST and alanine transaminase-ALT) enzymes were recorded in the hatchlings of Cyprinus carpio, Ctenopharyngodon idella, Labeo rohita and Cirrhinus mrigala after 7 and 14 days exposure and 10 days post exposure (recovery period) to sublethal concentrations (0.005, 0.01, 0.02 and 0.05 mg/L) of triclosan, a highly toxic and persistent biocide used in personal care products. Accumulation was maximum between 7-14 days at 0.01 mg/L for C. carpio and L. rohita but at 0.005 mg/L for C. idella and C. mrigala. No triclosan was observed at 0.005 mg/L in C. carpio and C. mrigala after recovery. Significant decline in protein, glutathione and acetylcholinesterase but increase in glutathione S-transferase, lactate dehydrogenase, aspartate transaminase, alanine transaminase, melondialdehyde and organic acids over control during exposure continued till the end of recovery period. Integrated biomarker response (IBR) analysis depicted higher star plot area for glutathione and glutathione S-transferase during initial 7 days of exposure, thereafter, during 7-14 days of exposure and the recovery period, higher star plot area was observed for acetylcholinesterase, aspartate transaminase, alanine transaminase and organic acids. Higher star plot area was observed for protein in all the species throughout the study. The study shows that L. rohita is most sensitive and glutathione, acetylcholinesterase, aspartate transaminase and alanine transaminase are the biomarkers for the toxicity of sublethal concentrations of TCS. Title: Differential effects of alprazolam against methylphenidate-induced neurobehavioral alterations Dutt M, Dharavath RN, Kaur T, Chopra K, Sharma S Ref: Physiol Behav, 222:112935, 2020 : PubMed ESTHER: Dutt_2020_Physiol.Behav_222_112935 PubMedSearch: Dutt 2020 Physiol.Behav 222 112935 PubMedID: 32413536 Abstract BACKGROUND: In the previous decade, abuse of several types of prescription drugs, particularly anxiolytics, opioid analgesics, and stimulants has increased significantly worldwide. Methylphenidate (MPH) and Alprazolam (ALZ) are extensively used drugs for the treatment of attention deficit hyperactivity disorder (ADHD) and anxiety disorders, respectively. However, these drugs have a high risk of being misused or abused alone, and their combination in some peculiar cases has shown their deleterious effects. In this study, we evaluated the extent of damage both these drugs (MPH and ALZ) may cause in the brain at different dosages. METHODS: Female Wistar rats were administered with MPH (10, 20, 40mg/kg) and ALZ (5, 10, 20mg/kg) alone and in combination. Following the treatment, neurobehavioral studies were conducted, and later brain tissue was removed for studying the extent of oxidative stress and inflammation in the hippocampus and cortex region of the brain. Further histopathological parameters, along with neurotransmitter levels, were also assessed. RESULTS: Both MPH and ALZ, in combination, enhanced oxidative stress, inflammation, and neurobehavioral alterations in a dose-dependent manner. These toxic effects were associated with histopathological alterations and neurotransmitters levels CONCLUSIONS: In this study, it is found that the combination of psychostimulant (MPH) and depressant (ALZ) tends to enhance toxicity in the brain, and their long-term usage is a significant public health concern. Therefore, their co-administration should be strictly monitored by medical practitioners, and under compulsive circumstances, their use must be restricted to lower doses. Title: Diapocynin, an NADPH oxidase inhibitor, counteracts diisopropylfluorophosphate-induced long-term neurotoxicity in the rat model Putra M, Gage M, Sharma S, Gardner C, Gasser G, Anantharam V, Thippeswamy T Ref: Annals of the New York Academy of Sciences, :, 2020 : PubMed ESTHER: Putra_2020_Ann.N.Y.Acad.Sci__ PubMedSearch: Putra 2020 Ann.N.Y.Acad.Sci PubMedID: 32037612 Abstract Organophosphate (OP) nerve agents are a threat to both the military and civilians. OP exposure causes cholinergic crisis and status epilepticus (SE) because of irreversible inhibition of acetylcholinesterase that can be life-threatening if left untreated. OP survivors develop long-term morbidity, such as cognitive impairment and motor dysfunction, because of oxidative stress and progressive neuroinflammation and neurodegeneration, which act as disease promoters. Current medical countermeasures (MCMs) do not mitigate these pathologies. Therefore, our goal was to target these disease promoters using diapocynin (DPO), an NADPH oxidase inhibitor, in addition to MCMs, in a rat diisopropylfluorophosphate (DFP) model. The DFP-intoxicated rats were treated with DPO (300 mg/kg, oral, six doses, 12-h intervals) or vehicle 2 h following behavioral SE termination with diazepam. The DPO treatment significantly rescued DFP-induced motor impairment and attenuated epileptiform spiking during the first 72 h after DFP exposure in severely seizing rats despite no difference in epileptiform spike rate between the vehicle and DPO groups in mild SE rats. DPO significantly reduced DFP-induced reactive astrogliosis, neurodegeneration, GP91(phox) , glutathiolated protein, serum nitrite, and proinflammatory cytokines and chemokines, such as interleukins (ILs) IL-1alpha, IL-6, IL-2, IL-17A, leptin, and IP-10, in the hippocampus. Collectively, these data support a neuroprotective role of DPO in an OP-induced neurotoxicity model. Title: Interaction of Synthesized Nitrogen enriched Graphene Quantum Dots with Novel Anti-Alzheimer's Drugs: Spectroscopic Insights Sharma S, Singh N, Nepovimova E, Korabecny J, Kuca K, Satnami ML, Ghosh KK Ref: J Biomol Struct Dyn, 38:1822, 2020 : PubMed ESTHER: Sharma_2020_J.Biomol.Struct.Dyn_38_1822 PubMedSearch: Sharma 2020 J.Biomol.Struct.Dyn 38 1822 PubMedID: 31096863 Abstract Alzheimer's disease (AD) is considered as one of widespread dementia with no approved diagnosis, cure or prevention. Currently, only symptomatic relief can be provided upon administration of anti-AD drugs generally belonging to a category of anticholinesterases and antagonists of N-methyl-D-aspartate (NMDA) receptors. In present investigation, a sensing platform has been designed for studying recently developed anti-AD drugs viz., PC-25 (N-(2-{4-[(4-bromophenyl)methyl]piperazin-1-yl}ethyl)-7-methoxy-1,2,3,4-tetrahydr oacridin-9-amine trihydrochloride), PC-37 (7-methoxy-N-(2-{4-[(3-methylphenyl)methyl]piperazin-1-yl}ethyl)-1,2,3,4-tetrahyd roacridin-9-amine trihydrochloride) and PC-48 (N-(2-{4-[(3-bromophenyl) methyl]piperazin-1-yl}ethyl)-7-methoxy-1,2,3,4-tetrahydroacridin-9-amine trihydrochloride) and two known standard tacrine (THA) and donepezil drugs for their estimation of in vitro potency towards AD using spectroscopic method. Anti-AD drugs have been accounted for individually with highly fluorescent nitrogen doped graphene quantum dots (NGQDs). The designed anti-AD drugs exerted the efficacy related to cholinergic hypothesis of AD. While the enzyme action is sensed by interacted species of NGQDs and acetylcholine, the fluorescence of NGQDs is quenched by the hydrolyzing action of acetylcholinesterase (AChE) enzyme but the lost fluorescence is recovered back upon addition of anti-AD drugs. These alterations in fluorescence of NGQDs are expected to have biological relevance akin to sensing. Moreover, these results advocate that out of all the drugs tested PC-37 displayed maximum inhibition efficiency. Our investigations suggest that synthesized drugs have the AD treating potential and can be entrusted in the near future for AD treatment. The validation parameters such as LOD, LOQ and recovery (%) were calculated in the range of 2.87 mM, 9.58 mM and 85-96%, respectively. Title: Mining of potential dipeptidyl peptidase-IV inhibitors as anti-diabetic agents using integrated in silico approaches Sharma S, Srivastava S, Shrivastava A, Malik R, Almalki F, Saifullah K, Alam MM, Shaqiquzzaman M, Ali S, Akhter M Ref: J Biomol Struct Dyn, 38:5349, 2020 : PubMed ESTHER: Sharma_2020_J.Biomol.Struct.Dyn_38_5349 PubMedSearch: Sharma 2020 J.Biomol.Struct.Dyn 38 5349 PubMedID: 31813365 Abstract The Dipeptidyl peptidase-IV (DPP-IV) family of receptors possesses a large binding cavity that imparts promiscuity for number of ligand binding which is not common to other receptors. This feature increases the challenge of using computational methods to identify DPP-IV inhibitors, therefore using both pharmacophore and structure based screening seems to be a reliable approach. Mining of novel DPP-IV inhibitors by integrating both of these in silico techniques was reported. Pharmacophore model (Model_008) obtained from structurally diverse reported compounds was used as a template for screening of MolMall database followed by structure based screening against PDB ID: 5T4E. After Absorption, Distribution, Metabolism and Excretion (ADME) analysis of shortlisted compounds, consensus docking and Molecular Mechanics/Generalized Born Surface Area (MM/GBSA) studies were carried out. The results of the docking studies obtained were comparable to that of the reference ligand. Out of nine hits identified, only one hit (ID MolMall-20062) was available which was procured through exchange program. Molecular Dynamic (MD) simulation studies of the procured hit revealed its good selectivity and stability in DPP-IV binding pocket and interactions observed with important amino acids viz., Trp629, Lys544 and Arg125. Biological testing of the compound MolMall-20062 showed promising DPP-IV inhibition activity with IC50: 6.2microM. Compound MolMall-20062 could be taken as a good lead for the development of DPP-IV inhibitors.Abbreviations: ADME: Absorption, Distribution, Metabolism and Excretion; ChEBI: Chemical Entities of Biological Interest; DPP-IV: Dipeptidyl peptidase IV; DISCOtech: Distance Comparisons; HTVS: High Throughput Virtual Screening; MD: Molecular Dynamics; MM-GBSA: Molecular Mechanics-Generalized Born Surface Area; OGTT: Oral Glucose Tolerance Test; PBVS: Pharmacophore Based Virtual Screening; PDB: Protein Data Bank; RMSD: Root Mean Square Deviation; ROC: Receiver Operating Characteristics; SP: Standard Precision; SBVS: Structure Based Virtual Screening; VS: Virtual Screening; XP: Extra Precision. Title: Effect of electromagnetic radiation on redox status, acetylcholine esterase activity and cellular damage contributing to the diminution of the brain working memory in rats Sharma S, Shukla S Ref: Journal of Chemical Neuroanatomy, :101784, 2020 : PubMed ESTHER: Sharma_2020_J.Chem.Neuroanat__101784 PubMedSearch: Sharma 2020 J.Chem.Neuroanat 101784 PubMedID: 32205214 Abstract Behavioral impairments are the most pragmatic outcome of long-term mobile uses but the underlying causes are still poorly understood. Therefore, the Aim of the present study to determine the possible mechanism of mobile induced behavioral alterations by observing redox status, cholinesterase activity, cellular, genotoxic damage and cognitive alterations in rat hippocampus. This study was carried out on 24 male Wistar rats, randomly divided into four groups (n = 6 in each group): group I consisted of sham-exposed (control) rats, group II-IV consisted of rats exposed to microwave radiation (900 MHz) at different time duration 1 h, 2 h, and 4 h respectively for 90 days. After 90 days of exposure, rats were assessing learning ability by using T-Maze. A significantly increased level of malondialdehyde (MDA) with concomitantly depleted levels of superoxide dismutase (SOD), catalase (CAT) and redox enzymes (GSH, GPX, GR, GST, G-6PDH) indicated an exposure of mobile emitted EMR induced oxidative stress by the depleted redox status of brain cells. The depletion in the acetylcholinesterase (AChE) level reveals altered neurotransmission in brain cells. Resultant cellular degeneration was also observed in the radiation-exposed hippocampus. Conclusively, the present study revealed that microwave radiation induces oxidative stress, depleted redox status, and causes DNA damage with the subsequent reduction in working memory in a time-dependent manner. This study provides insight over the associative reciprocity between redox status, cellular degeneration and reduced cholinergic activity, which presumably leads to the behavioral alterations following mobile emitted electromagnetic radiation. Title: In silico strategies for probing novel DPP-IV inhibitors as anti-diabetic agents Sharma S, Srivastav S, Singh G, Singh S, Malik R, Alam MM, Shaqiquzamman M, Ali S, Akhter M Ref: J Biomol Struct Dyn, :1, 2020 : PubMed ESTHER: Sharma_2020_J.Biomol.Struct.Dyn__1 PubMedSearch: Sharma 2020 J.Biomol.Struct.Dyn 1 PubMedID: 32248758 Abstract Identification of new DPP-IV inhibitors by integrating validated in silico approach is being presented herein. Novel hits were identified by combining pharmacophore and structure based virtual screening of ZINC and Knowledge Base in house database followed by ADME profiling, consensus docking studies. Six potential hits were identified and analysed for their synthetic accessibility score, novelty analysis and pan assay interference compounds filtration. Out of six, three hits viz., ZINC25060187, ZINC53746227 and KB-10 were analysed for stability studies using Molecular Mechanics/Generalized Born Surface Area (MM/GBSA) and Molecular Dynamics (MD) simulation. The simulation studies of the identified hits revealed that these hits have good selectivity and stability in DPP-IV binding pocket. Important interactions with amino acids viz., Tyr547, Glu205 and Glu206 similar to co-crystallized ligand were also observed. One of the hits viz., KB-10 was synthesized and evaluated for its biological potential. The compound KB-10 showed good DPP-IV inhibition in both in vitro and in vivo studies with IC50: 22.69 microM. This study supports the fact that these techniques hold potential for efficient screening of compounds with unknown affinity for DPP-IV that could serve as candidates for therapeutic development. Title: New coumarin-benzotriazole based hybrid molecules as inhibitors of acetylcholinesterase and amyloid aggregation Singh A, Sharma S, Arora S, Attri S, Kaur P, Kaur Gulati H, Bhagat K, Kumar N, Singh H and Mohinder Singh Bedi P <1 more author(s)> Singh A, Sharma S, Arora S, Attri S, Kaur P, Kaur Gulati H, Bhagat K, Kumar N, Singh H, Vir Singh J, Mohinder Singh Bedi P (- 1) Ref: Bioorganic & Medicinal Chemistry Lett, 30:127477, 2020 : PubMed ESTHER: Singh_2020_Bioorg.Med.Chem.Lett_30_127477 PubMedSearch: Singh 2020 Bioorg.Med.Chem.Lett 30 127477 PubMedID: 32781220 Abstract A novel series of triazole tethered coumarin-benzotriazole hybrids based on donepezil skeleton has been designed and synthesized as multifunctional agents for the treatment of Alzheimer's disease (AD). Among the synthesized compounds 13b showed most potent acetylcholinesterase (AChE) inhibition (IC(50) = 0.059 microM) with mixed type inhibition scenario. Structure-activity relationship revealed that three-carbon alkyl chain connecting coumarin and triazole is well tolerable for inhibitory potential. Hybrids obtained from 4-hydroxycoumarin and 1-benzotriazole were most potent AChE inhibitors. The inhibitory potential of all compounds against butyrylcholinesterase was also evaluated but all showed negligible activity suggesting that the hybrid molecules are selective AChE inhibitors. 13b (most potent AChE inhibitor) also showed copper-induced Abeta(1-42) aggregation inhibition (34.26% at 50 microM) and chelating properties for metal ions (Cu(2+), Fe(2+,) and Zn(2+)) involved in AD pathogenesis along with DNA protective potential against degenerative actions of OH radicals. Molecular modelling studies confirm the potential of 13b in blocking both PAS and CAS of AChE. In addition, interactions of 13b with Abeta(1-42) monomer are also streamlined. Therefore, hybrid 13b can act as an effective hit lead molecule for further development of selective AChE inhibitors as multifunctional anti-Alzheimer's agents. Title: Clitoria ternatea Mediated Synthesis of Graphene Quantum Dots for the Treatment of Alzheimer's Disease Tak K, Sharma R, Dave V, Jain S, Sharma S Ref: ACS Chem Neurosci, 11:3741, 2020 : PubMed ESTHER: Tak_2020_ACS.Chem.Neurosci_11_3741 PubMedSearch: Tak 2020 ACS.Chem.Neurosci 11 3741 PubMedID: 33119989 Abstract The main purpose of the present study was to synthesize graphene quantum dots (GQDs)from the flowers of Clitoria ternatea with the help of one-pot microwave-assisted green synthesis for the treatment of Alzheimer's disease. Further, the synthesized graphene quantum dots show a particle size of 10 nm +/-1.3, a PDI of 0.354 +/- 1.8, and a potential of -46 +/- 0.4, indicating the good stability of the quantum dots. With the help of scanning electron microscopy (SEM) and transfer electron microscopy (TEM) examination, the surface microscopic behavior of the synthesized quantum dots was determined. The presence of functional groups in the quantum dots was determined by Fourier-transform infrared spectroscopy (FTIR) study, the chemical state information on the sample was determined with the help of X-ray photoelectron spectroscopy (XPS), and the surface area of the dots was determined with the help of a surface area analyzer. With the help of a radial arm maze and water morris maze assay, the learning and memory capacity of the quantum dots was assessed, and the results show that the ctGQDs significantly decreased the transfer latency to reach the baited arm in 10.37 +/- 1.65 s or to the hidden platform in 18.42 +/- 0.99 s in 7 days. The synthesized quantum dots show more inhibition of the acetyl cholinesterase enzyme, i.e., 86.32 +/- 1.52%, as compared to that of pure donepezil, i.e., 72.46 +/- 2.21%. ctGQDs considerably increased the level of glutathione and protein and decreased the level of lipid peroxide and nitric oxide. The histopathological image of ctGQDs shows more preservation of small pyramidal cell and treats the disorganization of the cells. These results suggest that the quantum dots significantly crossed the blood-brain barrier since they were small in size and were effective in reducing Alzheimer-like symptoms in rodents, and thus, it can be concluded that Clitoria ternatea flowers can be used as an adjuvant in the treatment of Alzheimer's. Title: The effect of pyridostigmine on small intestinal bacterial overgrowth (SIBO) and plasma inflammatory biomarkers in HIV-associated autonomic neuropathies Robinson-Papp J, Nmashie A, Pedowitz E, George MC, Sharma S, Murray J, Benn EKT, Lawrence SA, Machac J and Morgello S <4 more author(s)> Robinson-Papp J, Nmashie A, Pedowitz E, George MC, Sharma S, Murray J, Benn EKT, Lawrence SA, Machac J, Heiba S, Kim-Schulze S, Navis A, Roland BC, Morgello S (- 4) Ref: J Neurovirol, 25:551, 2019 : PubMed ESTHER: Robinson-Papp_2019_J.Neurovirol_25_551 PubMedSearch: Robinson-Papp 2019 J.Neurovirol 25 551 PubMedID: 31098925 Inhibitor(s) related to this paper: Pyridostigmine Abstract Small intestinal bacterial overgrowth (SIBO) is common among patients with HIV-associated autonomic neuropathies (HIV-AN) and may be associated with increased bacterial translocation and elevated plasma inflammatory biomarkers. Pyridostigmine is an acetylcholinesterase inhibitor which has been used to augment autonomic signaling. We sought preliminary evidence as to whether pyridostigmine could improve proximal gastrointestinal motility, reduce SIBO, reduce plasma sCD14 (a marker of macrophage activation and indirect measure of translocation), and reduce the inflammatory cytokines IL-6 and TNFalpha in patients with HIV-AN. Fifteen participants with well-controlled HIV, HIV-AN, and SIBO were treated with 8 weeks of pyridostigmine (30 mg PO TID). Glucose breath testing for SIBO, gastric emptying studies (GES) to assess motility, plasma sCD14, IL-6, and TNFalpha, and gastrointestinal autonomic symptoms were compared before and after treatment. Thirteen participants (87%) experienced an improvement in SIBO following pyridostigmine treatment; with an average improvement of 50% (p = 0.016). There was no change in gastrointestinal motility; however, only two participants met GES criteria for gastroparesis at baseline. TNFalpha and sCD14 levels declined by 12% (p = 0.004) and 19% (p = 0.015), respectively; there was no significant change in IL-6 or gastrointestinal symptoms. Pyridostigmine may ameliorate SIBO and reduce levels of sCD14 and TNFalpha in patients with HIV-AN. Larger placebo-controlled studies are needed to definitively delineate how HIV-AN affects gastrointestinal motility, SIBO, and systemic inflammation in HIV, and whether treatment improves clinical outcomes. Title: Mobile phone induced cognitive and neurochemical consequences Sharma A, Sharma S, Shrivastava S, Singhal PK, Shukla S Ref: Journal of Chemical Neuroanatomy, :101684, 2019 : PubMed ESTHER: Sharma_2019_J.Chem.Neuroanat__101684 PubMedSearch: Sharma 2019 J.Chem.Neuroanat 101684 PubMedID: 31553920 Abstract With the rapid advances in technology, extensive use of mobile phones has increased the risk of health problems. This study was performed to find out the effect of mobile phone frequency on male Wistar rats. Animals were divided into two groups (n = 6 in each group). Group one was considered as control and group two (experimental group) was exposed to microwave radiation (2100 MHz) for 4 hours/day (5 days/week) for 3 months. Exposure of microwave radiation frequency showed significant alterations in cholinesterase activity, muscular strength, learning ability and anxiety. MWR exposure was also associated with significant alteration in the oxidative defense system and hippocampus degeneration. Histopathological observations clearly depicted the neural degeneration. Thus, it can be concluded that MWR significantly affects the central nervous system and may lead to many severe illnesses. This study may reveal a platform to understand its toxic effect and can further be used for amendment in current guidelines of mobile radiation. Title: The protective effect of alpha-lipoic acid against bisphenol A-induced neurobehavioral toxicity Khan J, Salhotra S, Ahmad S, Sharma S, Abdi SAH, Banerjee BD, Parvez S, Gupta S, Raisuddin S Ref: Neurochem Int, 118:166, 2018 : PubMed ESTHER: Khan_2018_Neurochem.Int_118_166 PubMedSearch: Khan 2018 Neurochem.Int 118 166 PubMedID: 29908256 Abstract Bisphenol A (BPA), a well-known xenoestrogen, is ubiquitously utilized in manufacturing of polycarbonated plastics. Convincing evidence suggests that BPA induces neurotoxicity and certain behavioral deficits. alpha-Lipoic acid (ALA) supplementation has shown protective effect against heart and liver diseases, diabetes, and neurological debility associated with aging. We studied the neuromodulatory effect of ALA against neurotoxicity of BPA in vitro in C8-D1A mouse astrocyte cell line and in vivo in C57BL/6J male mice. In vitro ALA (100muM) protected cells from BPA (30muM)-induced reactive oxygen species generation and increased activity of glial fibrillary acidic protein. ALA showed reduction in cell death in astrocytes treated with BPA. In vivo ALA (50mg/kg) increased the neurospecific acetylcholinesterase activity and decreased the monoamine oxidase activity altered by BPA exposure (10mg/kg, per os x 30 days). In addition to neuroprotective effects, ALA also showed protective effects against BPA-induced oxidative stress. We observed that ALA significantly replenished the declined neurobehavioral and cognitive performances, decreased muscle coordination and alerted short-term recognition memory in mice exposed to BPA. Our results suggest that ALA has a promising role in modulating BPA-induced neurotoxicity in C8-D1A mouse astrocyte cells as well as neurochemical and neurobehavioral deficits in C57BL/6J male mice and its antioxidant and free radical scavenging activities may in part be responsible for such an effect. Title: Rational Approaches, Design Strategies, Structure Activity Relationship and Mechanistic Insights for Esterase Inhibitors Singh H, Singh JV, Kaur N, Sanduja M, Singh G, Bedi PMS, Sharma S Ref: Mini Rev Med Chem, 18:837, 2018 : PubMed ESTHER: Singh_2018_Mini.Rev.Med.Chem_18_837 PubMedSearch: Singh 2018 Mini.Rev.Med.Chem 18 837 PubMedID: 28782481 Abstract BACKGROUND: Esterase is an enzyme that splits esters into an acid and alcohol. Varieties of esterases are present in human body to control diverse set of cellular processes and execute their specific functions. It can be seen that any increase in metabolites produced by these enzymes lead to severe pathological conditions like Alzheimer disease, hypercholesterolemia etc. Objective: Numerous esterase inhibitors have been developed and reported by the researchers around the globe, but not systematically summarized yet. Therefore, this assemblage focuses on various reported esterase inhibitors during recent past with detailed account of the design strategies employed for the synthesis of novel drug entities. The article also highlights the structure activity relationship along with mechanistic insights revealed during the biological evaluation of inhibitors as esterase inhibition. The interactions with the amino acid residues responsible for esterase inhibitory potential of molecules have also been discussed. This compilation will be of great interest for the researchers working in the area of esterase inhibitors. CONCLUSION: Rivastigmine derivatives (44-53), tacrine-piperazine hybrid (136), coumarin-benzofuran derivative (169), coumarin-benzylpiperidine hybrid (181) and phenylcinnamide derivative (220) found to be exerting cholinesterase inhibition with IC50 below the range of 1 nM. Whereas, flavone (258) has displayed anticholesterol esterase potential below 1 nM. Benzil like derivative, (273) has also been designed and reported to possess remarkable inhibitory potential (IC50 < 1 nM) against carboxylesterase. These representative results place them in forefront as potential future drug candidates to further develop potent and specific esterase inhibitors. Title: Involvement of glucose related energy crisis and endoplasmic reticulum stress: Insinuation of streptozotocin induced Alzheimer's like pathology Biswas J, Gupta S, Verma DK, Gupta P, Singh A, Tiwari S, Goswami P, Sharma S, Singh S Ref: Cell Signal, 42:211, 2017 : PubMed ESTHER: Biswas_2017_Cell.Signal_42_211 PubMedSearch: Biswas 2017 Cell.Signal 42 211 PubMedID: 29126976 Abstract The present study was conducted to correlate the cellular and molecular alterations in Alzheimer's pathology employing streptozotocin (STZ) induced experimental rat model. The STZ was administered in rat brain bilaterally by intracerebroventricular route using stereotaxic surgery followed by donepezil dosing. The Alzheimer's related pathological marker like acetylcholinesterase (AChE) activity, tau phosphorylation and amyloid aggregation were observed after STZ administration. STZ treatment showed decreased glucose and glucose transporters (GLUT) level along with augmented level of calcium in both cortical and hippocampal regions of rat brain. Increased calcium level may correlate with endoplasmic reticulum (ER) stress and significantly increased expression of ER stress markers like GRP78, GADD and caspase-12 were observed in STZ treated rat brain. Cellular communication was also affected by STZ administration as observed by increased expression connexin 43. With this view the activation of astrocytes and microglia was also assessed and observed by augmented GFAP and cd11b expression which were partially inhibited with donepezil treatment. The significantly increased level of degenerating neurons, caspase-3 and DNA fragmentation was also observed in rat brain regions which were not inhibited with donepezil treatment and validating the clinical observations. In conclusion, study indicated the STZ induced occurrence of Alzheimer's pathology. Further, STZ administration also caused depleted glucose level, inhibited mitochondrial activity, augmented calcium levels, ER stress, altered cellular communication and neuronal death which were partially attenuated with donepezil treatment. Title: Bioresolution of racemic phenyl glycidyl ether by a putative recombinant epoxide hydrolase from Streptomyces griseus NBRC 13350 Saini P, Kumar N, Wani SI, Sharma S, Chimni SS, Sareen D Ref: World J Microbiol Biotechnol, 33:82, 2017 : PubMed ESTHER: Saini_2017_World.J.Microbiol.Biotechnol_33_82 PubMedSearch: Saini 2017 World.J.Microbiol.Biotechnol 33 82 PubMedID: 28378221 Gene_locus related to this paper: strgg-b1vn62 Abstract In order to produce enantiomerically pure epoxides for the synthesis of value-added chemicals, a novel putative epoxide hydrolase (EH) sgeh was cloned and overexpressed in pET28a/Escherichia coli BL21(DE3). The 1047 bp sgeh gene was mined from Streptomyces griseus NBRC 13350 genome sequence. The recombinant hexahistidyl-tagged SGEH was purified (16.6-fold) by immobilized metal-affinity chromatography, with 90% yield as a homodimer of 100 kDa. The recombinant E. coli whole cells overexpressing SGEH could kinetically resolve racemic phenyl glycidyl ether (PGE) into (R)-PGE with 98% ee, 40% yield, and enantiomeric ratio (E) of 20. This was achieved under the optimized reaction conditions i.e. cell/substrate ratio of 20:1 (w/w) at pH 7.5 and 20 degrees C in 10% (v/v) dimethylformamide (DMF) in a 10 h reaction. 99% enantiopure (R)-PGE was obtained when the reaction time was prolonged to 12 h with a yield of 34%. In conclusion, an economically viable and environment friendly green process for the production of enantiopure (R)-PGE was developed by using wet cells of E. coli expressing recombinant SGEH. Title: Chlorpyrifos pollution: its effect on brain acetylcholinesterase activity in rat and treatment of polluted soil by indigenous Pseudomonas sp Sharma S, Singh PB, Chadha P, Saini HS Ref: Environ Sci Pollut Res Int, 24:381, 2017 : PubMed ESTHER: Sharma_2017_Environ.Sci.Pollut.Res.Int_24_381 PubMedSearch: Sharma 2017 Environ.Sci.Pollut.Res.Int 24 381 PubMedID: 27722883 Abstract The study was aimed to evaluate the levels of chlorpyrifos (CPF) pollution in agricultural soil of Punjab, India, its detrimental effects on acetylcholinesterase (AChE) activity in rat brain and bioremediation of soils polluted with CPF using indigenous and adapted bacterial lab isolate. The analysis revealed that soil samples of Bathinda and Amritsar regions are highly contaminated with chlorpyrifos showing 19 to 175 mg/kg concentrations of CPF. The non-targeted animals may get poisoned with CPF by its indirect dermal absorption, inhalation of toxic fumes and regular consumption of soiled food grains. The study indicated that even the lowermost concentrations of CPF, 19 and 76 mg/kg of soil found in the Amritsar and Bathinda regions respectively can significantly inhibit the AChE activity in rat brain within 24 h of its treatment. This represents the antagonistic effect of CPF on AChE which is a prime neurotransmitter present in all living beings including humans. In light of this, an attempt was made to remediate the polluted soil, a major reservoir of CPF, using Pseudomonas sp. (ChlD), an indigenous bacterial isolate. The culture efficiently degraded 10 to 100 mg/kg chlorpyrifos supplemented in the soil and utilized it as sole source of carbon and energy for its growth. Thus, this study provides a detailed insight regarding the level of CPF pollution in Punjab, its detrimental effects on mammals and bio-based solution to remediate the sites polluted with CPF. Title: Annotation of gene sequence and protein structure of brinjal EDS1 Sharma S, Jaiswal S, Archak S Ref: Bioinformation, 13:54, 2017 : PubMed ESTHER: Sharma_2017_Bioinformation_13_54 PubMedSearch: Sharma 2017 Bioinformation 13 54 PubMedID: 28584443 Abstract Enhanced Disease Susceptibility1 (EDS1) is a nucleo-cytoplasmic protein, known to be a key regulator of plant basal defense and effector-triggered immunity. Sequence of a single copy brinjal EDS1 gene (SmEDS1) was mined from draft brinjal genome assembly. The extracted sequence was found to be incomplete and polished with the help of transcriptome sequence data. Full-length SmEDS1 gene is 4.5kb long having three exons that coded for 1.8kb mRNA. SmEDS1 protein is a 602 amino acid long protein consisting of Lipase3 and EP domain regions. Predicted tertiary structure of SmEDS1 using homology modelling had a mass of 68.8kD and was made of 10 strands, 26 alpha helices, five 310 helices and 43 beta turns. Phylogenetic analysis based on protein sequence grouped the species in clades defined by botanical family suggesting that EDS1 protein has evolved through the speciation process. Phylogenetic tree based on EDS1 structures grouped Solanum species of American origin (tomato, wild tomato and potato) together but brinjal EDS1 (Asiatic origin) occupied a unique position. In silico information generated in this study is expected to be the first step toward cloning and expression analysis of SmEDS1 gene. Title: Benzoflavones as cholesterol esterase inhibitors: Synthesis, biological evaluation and docking studies Singh H, Singh JV, Gupta MK, Singh P, Sharma S, Nepali K, Bedi PM Ref: Bioorganic & Medicinal Chemistry Lett, 27:850, 2017 : PubMed ESTHER: Singh_2017_Bioorg.Med.Chem.Lett_27_850 PubMedSearch: Singh 2017 Bioorg.Med.Chem.Lett 27 850 PubMedID: 28117203 Abstract A library of forty 7,8-benzoflavone derivatives was synthesized and evaluated for their inhibitory potential against cholesterol esterase (CEase). Among all the synthesized compounds seven benzoflavone derivatives (A-7, A-8, A-10, A-11, A-12, A-13, A-15) exhibited significant inhibition against CEase in in vitro enzymatic assay. Compound A-12 showed the most promising activity with IC50 value of 0.78nM against cholesterol esterase. Enzyme kinetic studies carried out for A-12, revealed its mixed-type inhibition approach. Molecular protein-ligand docking studies were also performed to figure out the key binding interactions of A-12 with the amino acid residues of the enzyme's active site. The A-12 fits well at the catalytic site and is stabilized by hydrophobic interactions. It completely blocks the catalytic assembly of CEase and prevents it to participate in ester hydrolysis mechanism. The favorable binding conformation of A-12 suggests its prevailing role as CEase inhibitor. Title: Neurokinin-1 receptor inhibition reverses ischaemic brain injury and dementia in bilateral common carotid artery occluded rats: possible mechanisms Kaur J, Sharma S, Sandhu M Ref: Inflammopharmacology, 24:133, 2016 : PubMed ESTHER: Kaur_2016_Inflammopharmacology_24_133 PubMedSearch: Kaur 2016 Inflammopharmacology 24 133 PubMedID: 27380492 Abstract BACKGROUNDS: Increase in SP release as a function of hypoxia of the rat carotid body is a tissue response to ischemia that leads to neurogenic inflammation and cognitive deficits. Substance P-mediated inflammation is reported to attenuate the neuroprotective PPAR-gamma. This study was undertaken to investigate the effect of aprepitant, a substance P-NK1 receptor antagonist in bilateral carotid artery occlusion (BCCAO)-induced ischaemic brain injury and vascular dementia. Title: Are PECTIN ESTERASE INHIBITOR Genes Involved in Mediating Resistance to Rhynchosporium commune in Barley? Marzin S, Hanemann A, Sharma S, Hensel G, Kumlehn J, Schweizer G, Roder MS Ref: PLoS ONE, 11:e0150485, 2016 : PubMed ESTHER: Marzin_2016_PLoS.One_11_e0150485 PubMedSearch: Marzin 2016 PLoS.One 11 e0150485 PubMedID: 26937960 Abstract A family of putative PECTIN ESTERASE INHIBITOR (PEI) genes, which were detected in the genomic region co-segregating with the resistance gene Rrs2 against scald caused by Rhynchosporium commune in barley, were characterized and tested for their possible involvement in mediating resistance to the pathogen by complementation and overexpression analysis. The sequences of the respective genes were derived from two BAC contigs originating from the susceptible cultivar 'Morex'. For the genes HvPEI2, HvPEI3, HvPEI4 and HvPEI6, specific haplotypes for 18 resistant and 23 susceptible cultivars were detected after PCR-amplification and haplotype-specific CAPS-markers were developed. None of the tested candidate genes HvPEI2, HvPEI3 and HvPEI4 alone conferred a high resistance level in transgenic over-expression plants, though an improvement of the resistance level was observed especially with OE-lines for gene HvPEI4. These results do not confirm but also do not exclude an involvement of the PEI gene family in the response to the pathogen. A candidate for the resistance gene Rrs2 could not be identified yet. It is possible that Rrs2 is a PEI gene or another type of gene which has not been detected in the susceptible cultivar 'Morex' or the full resistance reaction requires the presence of several PEI genes. Title: Genomic Resource of Rice Seed Associated Bacteria Midha S, Bansal K, Sharma S, Kumar N, Patil PP, Chaudhry V, Patil PB Ref: Front Microbiol, 6:1551, 2016 : PubMed ESTHER: Midha_2016_Front.Microbiol_6_1551 PubMedSearch: Midha 2016 Front.Microbiol 6 1551 PubMedID: 26793183 Gene_locus related to this paper: 9mico-a0a147dug2, 9micc-a0a147e820, 9mico-a0a147eme3, micte-a0a147ewf8, micte-a0a147f3r1, micte-a0a147f975, micte-a0a147f9m3, micte-a0a147f9s1, 9burk-a0a147gqr5, 9burk-a0a147h2a8, 9burk-a0a147h429, 9sphn-a0a147hsu0, 9sphn-a0a147i6k5, 9sphn-a0a147is40, 9sphn-a0a147it61, 9sphn-a0a147ejn4, 9burk-a0a147gmx1, 9psed-a0a147gux9, 9sphn-a0a147i2i3, 9sphn-a0a147i7r1, 9sphn-a0a147i964, 9sphn-a0a147ikp1, 9rhiz-a0a175ree3, 9sphn-a0a147eck7, 9sphn-a0a147iln2 Title: Alzheimer's disease like pathology induced six weeks after aggregated amyloid-beta injection in rats: increased oxidative stress and impaired long-term memory with anxiety-like behavior Sharma S, Verma S, Kapoor M, Saini A, Nehru B Ref: Neurol Res, :1, 2016 : PubMed ESTHER: Sharma_2016_Neurol.Res__1 PubMedSearch: Sharma 2016 Neurol.Res 1 PubMedID: 27431920 Abstract OBJECTIVES: Amyloid-beta (Abeta) peptide deposition into insoluble plaques is a pathological hallmark of Alzheimer's disease (AD), but soluble oligomeric Abeta is considered to be more potent and has been hypothesized to directly impair learning and memory. Also, evidences from some clinical studies indicated that Abeta oligomer formation is the major cause for early AD onset. However, the biochemical mechanism involved in the oligomer-induced toxicity is not very well addressed. So, thise present study was undertaken to study the effects of single intracerebroventricular (icv) injection of protofibrillar Abeta 1-42 on the behavioral and biochemical profile in rats. Title: Ascorbyl palmitate synthesis in an organic solvent system using a Celite-immobilized commercial lipase (Lipolase 100L) Sharma S, Kanwar K, Kanwar SS Ref: 3 Biotech, 6:183, 2016 : PubMed ESTHER: Sharma_2016_3.Biotech_6_183 PubMedSearch: Sharma 2016 3.Biotech 6 183 PubMedID: 28330255 Abstract Ascorbyl palmitate was synthesized using a Celite-immobilized commercial lipase (Lipolase 100L) in dimethylsulfoxide (DMSO) as an organic solvent system. Lipase immobilized by surface adsorption onto Celite 545 matrix and subsequently exposed to 1 % glutaraldehyde showed 75 % binding of protein. The Celite-bound lipase was optimally active at 75 degreesC and pH 8.5 under shaking and showed maximum hydrolytic activity toward p-NPP as a substrate. The bound lipase was found to be stimulated only in the presence of Al(3+) and EDTA. All surfactants (Tween-20, Tween-80 and Triton X-100) had an inhibitory effect on lipase activity. The optimization of various reaction conditions of ascorbyl palmitate was achieved considering one factor at a time. The esterification of ascorbic acid and palmitic acid was carried out with 1 M ascorbic acid and 2.5 M palmitic acid in DMSO at 75 degreesC for 18 h under shaking (120 rpm). Molecular sieves had an important effect on the ester synthesis resulting in an enhanced yield. The by-product (H(2)O) produced in the reaction was scavenged by the molecular sieves (20 mg/ml) added in the reaction mixture which enhanced the ester yield to 80 %. The characterization of synthesized ester was done through FTIR spectroscopy. Title: Use of ligand-based pharmacophore modeling and docking approach to find novel acetylcholinesterase inhibitors for treating Alzheimer's Dhanjal JK, Sharma S, Grover A, Das A Ref: Biomed Pharmacother, 71:146, 2015 : PubMed ESTHER: Dhanjal_2015_Biomed.Pharmacother_71_146 PubMedSearch: Dhanjal 2015 Biomed.Pharmacother 71 146 PubMedID: 25960230 Abstract Alzheimer's disease is a neurological disorder in which the patient suffers from memory loss and impaired cognitive abilities. Though the main cause of the disease is not yet known, depletion of neurotransmitter at synaptic junctions, accumulation of insoluble beta amyloid plaques and neurofibrillary tangles are the main pathologies associated with it. The FDA approved drugs for alzheimer's belong to the category of acetylcholinesterase inhibitors. But most of the drugs have been observed to be associated with adverse side effects. In this study, we have developed a pharmacophore (responsible for interaction with acetylcholinesterase active site) based on the already existing drugs and drug candidates. This pharmacophore was used to search for novel AChE inhibitors with altogether different chemical scaffold using high throughput virtual screening and docking studies. Finally, we have reported two compounds, OPA and OMT, which possess high affinity for catalytic site of AChE enzyme and thus, can be considered as potential AChE inhibitors for the symptomatic treatment of Alzheimer's. Title: Effect of Licofelone-A Dual COX/5-LOX Inhibitor in Intracerebroventricular Streptozotocin-Induced Behavioral and Biochemical Abnormalities in Rats Kumar A, Sharma S, Prashar A, Deshmukh R Ref: Journal of Molecular Neuroscience, 55:749, 2015 : PubMed ESTHER: Kumar_2015_J.Mol.Neurosci_55_749 PubMedSearch: Kumar 2015 J.Mol.Neurosci 55 749 PubMedID: 25204299 Abstract The present study was designed to investigate the effect of licofelone-a dual cyclooxygenase/5-lipoxygenase (COX/5-LOX) inhibitor in intracerebroventricular streptozotocin (ICV-STZ)-induced cognitive deficit and biochemical abnormalities in rats. ICV-STZ is a widely used model of sporadic Alzheimer's disease. In this study, STZ was administered intracerebroventricular (ICV)-bilaterally 3 mg/kg in rats. The STZ-injected rats were treated with different doses of licofelone (2.5, 5, and 10 mg/kg, p.o.) for 21 days. Cognitive functions were assessed by using Morris water maze and passive avoidance task. Levels of malondialdehyde (MDA), nitrite, reduced glutathione (GSH), and acetylcholinesterase (AChE) activity were determined to check oxidative stress and cholinergic function. Cytokine levels (IL-1beta and TNF-alpha) were also determined as markers of neuroinflammation. Administration of STZ caused a significant increase in AChE activity and cognitive dysfunction. Increased oxidative stress and the proinflammatory cytokine levels were also observed following STZ administration in rats. Licofelone treatment attenuated STZ-induced cholinergic hypofunction and cognitive deficit in rats. In addition, licofelone attenuated STZ-induced oxidative stress and elevated cytokine levels. The cognitive enhancement following licofelone administration in STZ rats may be due to its ability to restore cholinergic functions or its antioxidant activity. These observed results suggest the therapeutic potential of dual COX/5-LOX inhibitors in neurodegenerative disorders associated with oxidative stress and cognitive impairment. Title: Enhancement of stability of a lipase by subjecting to three phase partitioning (TPP): structures of native and TPP-treated lipase from Thermomyces lanuginosa Kumar M, Mukherjee J, Sinha M, Kaur P, Sharma S, Singh TP, Gupta MN Ref: Sustain Chem Process, 3:14, 2017 : PubMed ESTHER: Kumar_2015_Sustain.Chem.Process_3_14 PubMedSearch: Kumar 2015 Sustain.Chem.Process 3 14 Gene_locus related to this paper: humla-1lipa Inhibitor(s) related to this paper: Laurate Abstract Background The lipase enzyme converts long chain acyltriglycerides into di- and monoglycerides, glycerol and fatty acids. The catalytic site in lipase is situated deep inside the molecule. It is connected through a tunnel to the surface of the molecule. In the unbound state under aqueous conditions, the tunnel remains closed. The tunnel can be opened when the enzyme is exposed to a lipid bilayer or a detergent or many hydrophobic/hydrophilic surfaces. Results In the present study, the lipase was subjected to three-phase partitioning (TPP) which consisted of mixing in tert-butanol and ammonium sulphate to the solution of lipase in the aqueous buffer. The enzyme formed an interfacial precipitate between the tert-butanol rich and water rich phases. The stability of the enzyme subjected to TPP was found to be higher (Tm of 80 C) than the untreated enzyme (Tm of 77 C). The activity of the enzyme subjected to TPP (3.3 U/mg) was nearly half of that of the untreated one (5.8 U/mg). However, the activity of the treated enzyme was higher (17.8 U/mg) than the untreated one (8.6 U/mg) when a detergent was incorporated in the assay buffer. Conclusions The structure determination showed that the substrate binding site in the treated enzyme was more tightly closed than that of the untreated protein. Title: Discovery of a marine-derived bis-indole alkaloid fascaplysin, as a new class of potent P-glycoprotein inducer and establishment of its structure-activity relationship Manda S, Sharma S, Wani A, Joshi P, Kumar V, Guru SK, Bharate SS, Bhushan S, Vishwakarma RA and Bharate SB <1 more author(s)> Manda S, Sharma S, Wani A, Joshi P, Kumar V, Guru SK, Bharate SS, Bhushan S, Vishwakarma RA, Kumar A, Bharate SB (- 1) Ref: Eur Journal of Medicinal Chemistry, 107:1, 2015 : PubMed ESTHER: Manda_2015_Eur.J.Med.Chem_107_1 PubMedSearch: Manda 2015 Eur.J.Med.Chem 107 1 PubMedID: 26560048 Abstract The screening of IIIM natural products repository for P-gp modulatory activity in P-gp over-expressing human adenocarcinoma LS-180 cells led to the identification of 7 natural products viz. withaferin, podophyllotoxin, 3-demethylcolchicine, agnuside, reserpine, seseberecine and fascaplysin as P-gp inducers. Fascaplysin (6a), a marine-derived bis-indole alkaloid, was the most potent among all of them, showing induction of P-gp with EC50 value of 25 nM. P-gp induction is one of the recently targeted strategy to increase amyloid-beta clearance from Alzheimer brains. Thus, we pursued a medicinal chemistry of fascaplysin to establish its structure-activity relationship for P-gp induction activity. Four series of analogs viz. substituted quaternary fascaplysin analogs, D-ring opened quaternary analogs, D-ring opened non-quaternary analogs, and beta-carbolinium analogs were synthesized and screened for P-gp induction activity. Among the total of 48 analogs screened, only quaternary nitrogen containing analogs 6a-g and 10a, 10h-l displayed promising P-gp induction activity; whereas non-planar non-quaternary analogs 9a-m, 13a-n, 15a-h were devoid of this activity. The P-gp induction activity of best compounds was then confirmed by western-blot analysis, which indicated that fascaplysin (6a) along with 4,5-difluoro analog of fascaplysin 6f and D-ring opened analog 10j displayed 4-8 fold increase in P-gp expression in LS-180 cells at 1 muM. Additionally, compounds 6a and 6f also showed inhibition of acetylcholinestease (AChE), an enzyme responsible for neuronal loss in Alzheimer's disease. Thus, fascaplysin and its analogs showing promising P-gp induction along with AChE inhibition at 1 muM, with good safety window (LS-180: IC50 > 10 muM, hGF: 4 muM), clearly indicates their promise for development as an anti-Alzheimer agent. Title: Gallic acid-based alkyl esters synthesis in a water-free system by celite-bound lipase of Bacillus licheniformis SCD11501 Sharma S, Kanwar SS, Dogra P, Chauhan GS Ref: Biotechnol Prog, 31:715, 2015 : PubMed ESTHER: Sharma_2015_Biotechnol.Prog_31_715 PubMedSearch: Sharma 2015 Biotechnol.Prog 31 715 PubMedID: 25737230 Abstract Gallic acid (3, 4, 5- trihydroxybenzoic acid) is an important antioxidant, anti-inflammatory, and radical scavenging agent. In the present study, a purified thermo-tolerant extra-cellular lipase of Bacillus licheniformis SCD11501 was successfully immobilized by adsorption on Celite 545 gel matrix followed by treatment with a cross-linking agent, glutaraldehyde. The celite-bound lipase treated with glutaraldehyde showed 94.8% binding/retention of enzyme activity (36 U/g; specific activity 16.8 U/g matrix; relative increase in enzyme activity 64.7%) while untreated matrix resulted in 88.1% binding/retention (28.0 U/g matrix; specific activity 8.5 U/g matrix) of lipase. The celite-bound lipase was successfully used to synthesis methyl gallate (58.2%), ethyl gallate (66.9%), n-propyl gallate (72.1%), and n-butyl gallate (63.8%) at 55(o) C in 10 h under shaking (150 g) in a water-free system by sequentially optimizing various reaction parameters. The low conversion of more polar alcohols such as methanol and ethanol into their respective gallate esters might be due to the ability of these alcohols to severely remove water from the protein hydration shell, leading to enzyme inactivation. Molecular sieves added to the reaction mixture resulted in enhanced yield of the alkyl ester(s). The characterization of synthesised esters was done through fourier transform infrared (FTIR) spectroscopy and (1) H NMR spectrum analysis. Title: Beneficial Effect of Protein Tyrosine Phosphatase Inhibitor and Phytoestrogen in Dyslipidemia-Induced Vascular Dementia in Ovariectomized Rats Verma A, Sharma S Ref: J Stroke Cerebrovasc Dis, 24:2434, 2015 : PubMed ESTHER: Verma_2015_J.Stroke.Cerebrovasc.Dis_24_2434 PubMedSearch: Verma 2015 J.Stroke.Cerebrovasc.Dis 24 2434 PubMedID: 26324516 Abstract BACKGROUND: Estrogen deficiency and increase in protein tyrosine phosphatase (PTPase) activity may be a key mechanism in postmenopausal dyslipidemia-induced vascular dysfunction and dementia. Thus, the present study has been designed to investigate the effect of biochanin A (BCA, a phytoestrogen) and sodium orthovanadate (SOV), an inhibitor of PTPase in dyslipidemia-induced vascular dementia in ovariectomized rats. Title: Toxic effects of lead exposure in rats: involvement of oxidative stress, genotoxic effect, and the beneficial role of N-acetylcysteine supplemented with selenium Sharma S, Raghuvanshi BP, Shukla S Ref: J Environ Pathol Toxicol Oncol, 33:19, 2014 : PubMed ESTHER: Sharma_2014_J.Environ.Pathol.Toxicol.Oncol_33_19 PubMedSearch: Sharma 2014 J.Environ.Pathol.Toxicol.Oncol 33 19 PubMedID: 24579807 Abstract This study was carried out to investigate the in vivo protective role of N-acetylcysteine (NAC) per se, along with selenium (Se), against lead-induced hepatic, nephritic-oxidative, and neuronal-oxidative damage in rats. Lead acetate at a dose of 50 mg/kg body weight administered intraperitoneally for 3 days was preferred as the source of lead. Various oxidative stress markers such as reduced glutathione content, lipid peroxidation, superoxide dismutase, and catalase were measured to determine the degree of oxidative damage and healing due to NAC (50 mg/kg body weight administered orally) and Se (0.5 mg/kg body weight administered orally) and were studied along with the activities of enzymes such as transaminases (aspartate aminotransferase/alanine aminotransferase), delta-aminolevulinic acid dehydratase, delta-aminolevulinic acid synthase, and acetyl cholinesterase activity. The genotoxic effect of lead also was studied in terms of DNA damage using comet assay. The effect of lead was studied in blood biochemical variables such as cholesterol, triglycerides, urea, uric acid, and creatinine. Our data suggest that supplementation of Se with NAC can improve the lead-induced biochemical oxidative stress in blood and tissue, the burden of lead on the body, and molecular alterations by recoupment in mean DNA damage. Title: Organic Solvent Tolerant Lipases and Applications Sharma S, Kanwar SS Ref: ScientificWorldJournal, 2014:625258, 2014 : PubMed ESTHER: Sharma_2014_ScientificWorldJournal_2014_625258 PubMedSearch: Sharma 2014 ScientificWorldJournal 2014 625258 PubMedID: 24672342 Abstract Lipases are a group of enzymes naturally endowed with the property of performing reactions in aqueous as well as organic solvents. The esterification reactions using lipase(s) could be performed in water-restricted organic media as organic solvent(s) not only improve(s) the solubility of substrate and reactant in reaction mixture but also permit(s) the reaction in the reverse direction, and often it is easy to recover the product in organic phase in two-phase equilibrium systems. The use of organic solvent tolerant lipase in organic media has exhibited many advantages: increased activity and stability, regiospecificity and stereoselectivity, higher solubility of substrate, ease of products recovery, and ability to shift the reaction equilibrium toward synthetic direction. Therefore the search for organic solvent tolerant enzymes has been an extensive area of research. A variety of fatty acid esters are now being produced commercially using immobilized lipase in nonaqueous solvents. This review describes the organic tolerance and industrial application of lipases. The main emphasis is to study the nature of organic solvent tolerant lipases. Also, the potential industrial applications that make lipases the biocatalysts of choice for the present and future have been presented. Title: Neuroprotective role of Indirubin-3'-monoxime, a GSKbeta inhibitor in high fat diet induced cognitive impairment in mice Sharma S, Taliyan R Ref: Biochemical & Biophysical Research Communications, 452:1009, 2014 : PubMed ESTHER: Sharma_2014_Biochem.Biophys.Res.Commun_452_1009 PubMedSearch: Sharma 2014 Biochem.Biophys.Res.Commun 452 1009 PubMedID: 25234596 Abstract Recent studies have highlighted that diabetes mellitus (DM) is a strong risk factor for Alzheimer's disease (AD). Insulin resistance and/or hyperinsulinemia is one of the main characteristics of type 2 DM. Numerous epidemiological studies have demonstrated that insulin resistance contributes to AD pathogenesis. However the molecular mechanisms of association between these still remain elusive. Among the various possible mechanisms, the GSK-3beta activity has been reported to be impaired in insulin-resistance, type 2 DM and AD. Thus, the present study was designed to explore the neuroprotective role of GSK3 beta inhibitor, Indirubin-3'-monoxime (IMX) in insulin resistance induced cognitive impairment. Further, we have explored the possible molecular mechanism involved in cognitive impairment associated with insulin resistance. The mice subjected to high fat diet exhibited characteristic features of insulin resistance viz. increased serum glucose, triglycerides, cholesterol, insulin levels and impaired spatial learning and memory ability along with reduced brain insulin level, elevated oxidative stress and acetylcholinesterase (AChE) activity. The observed changes occurred concurrently with reduced brain derived neurotrophic factor. In contrast, the mice treated with IMX showed a significant reduction in plasma glucose, triglycerides, cholesterol, insulin levels and improvement in learning and memory performance, attenuated the oxidative stress and AChE activity. Moreover, IMX dose dependently augment the brain insulin and BDNF levels in HFD fed mice. Based upon these findings it could be suggested that GSK3 beta inhibition could prove to be beneficial in insulin resistance induced cognitive deficit and this neuroprotection could be the result of enhanced BDNF based synaptic plasticity. Title: Assessment of Serum Cholinesterase in Rural Punjabi Sprayers Exposed to a Mixture of Pesticides Dhalla AS, Sharma S Ref: Toxicol Int, 20:154, 2013 : PubMed ESTHER: Dhalla_2013_Toxicol.Int_20_154 PubMedSearch: Dhalla 2013 Toxicol.Int 20 154 PubMedID: 24082509 Abstract Serum cholinesterase (SChE) activity is considered as a biomarker and is also taken as an exposure index to assess the low level, chronic residue exposures among sprayers. Thus, cholinesterase activity was studied in the professional rural Punjabi sprayers of Bathinda district in Punjab. This study was made to estimate the irregularities in the level of cholinesterase according to multiple pesticides used by sprayers, exposure periods, age, and body mass index (BMI) of the sprayers. The data generated was statistically analyzed by applying Student's 't' test and one-way analysis of variance. A positive correlation was found between SChE activity and years of exposure and a significant reduction in SChE activity was observed in younger population. Again, a positive correlation was seen between BMI and SChE inhibition. Title: Knowledge, attitude and practices of pesticide use and acetylcholinesterase depression among farm workers in Nepal Atreya K, Kumar Sitaula B, Overgaard H, Man Bajracharya R, Sharma S Ref: Int J Environ Health Res, 22:401, 2012 : PubMed ESTHER: Atreya_2012_Int.J.Environ.Health.Res_22_401 PubMedSearch: Atreya 2012 Int.J.Environ.Health.Res 22 401 PubMedID: 22273496 Abstract Assessing erythrocyte acetylcholinesterase AChE activity in farm workers across agricultural seasons can be used to monitor risks of pesticide exposure We surveyed a total of 403 households in Nepal and adopted the Test-mate ChE Cholinesterase Test System to monitor AChE activity across season on the 127 individuals of the sampled households The study aims to i document knowledge and practices of pesticide use among farmers and ii present the relationship between farmers reported acute health symptoms and erythrocyte acetylcholinesterase depression We found low levels of pesticide use hygiene and high levels of individuals knowledge on the local environmental impacts of pesticide use Safety measures taken against potential risks of pesticides exposure were inadequate Exposure to organophosphates significantly reduced AChE activity across season but was not sufficient enough to claim clinical symptoms whereas exposure to the pyrethroid insecticides and fungicides were sufficient enough to claim acute symptoms of poisoning. Title: Expression, purification and crystallization of human prolylcarboxypeptidase. Abeywickrema PD, Patel SB, Byrne NJ, Diehl RE, Hall DL, Ford RE, Rickert KW, Reid JC, Shipman JM and Sharma S <5 more author(s)> Abeywickrema PD, Patel SB, Byrne NJ, Diehl RE, Hall DL, Ford RE, Rickert KW, Reid JC, Shipman JM, Geissler WM, Pryor KD, SinhaRoy R, Soisson SM, Lumb KJ, Sharma S (- 5) Ref: Acta Crystallographica Sect F Struct Biol Cryst Commun, 66:702, 2010 : PubMed ESTHER: Abeywickrema_2010_Acta.Crystallogr.Sect.F.Struct.Biol.Cryst.Commun_66_702 PubMedSearch: Abeywickrema 2010 Acta.Crystallogr.Sect.F.Struct.Biol.Cryst.Commun 66 702 PubMedID: 20516604 Gene_locus related to this paper: human-PRCP Abstract Prolylcarboxypeptidase (PrCP) is a lysosomal serine carboxypeptidase that cleaves a variety of C-terminal amino acids adjacent to proline and has been implicated in diseases such as hypertension and obesity. Here, the robust production, purification and crystallization of glycosylated human PrCP from stably transformed CHO cells is described. Purified PrCP yielded crystals belonging to space group R32, with unit-cell parameters a = b = 181.14, c = 240.13 A, that diffracted to better than 2.8 A resolution. Title: Structural definition and substrate specificity of the S28 protease family: the crystal structure of human prolylcarboxypeptidase Soisson SM, Patel SB, Abeywickrema PD, Byrne NJ, Diehl RE, Hall DL, Ford RE, Reid JC, Rickert KW and Lumb KJ <2 more author(s)> Soisson SM, Patel SB, Abeywickrema PD, Byrne NJ, Diehl RE, Hall DL, Ford RE, Reid JC, Rickert KW, Shipman JM, Sharma S, Lumb KJ (- 2) Ref: BMC Struct Biol, 10:16, 2010 : PubMed ESTHER: Soisson_2010_BMC.Struct.Biol_10_16 PubMedSearch: Soisson 2010 BMC.Struct.Biol 10 16 PubMedID: 20540760 Gene_locus related to this paper: human-PRCP Abstract BACKGROUND: The unique S28 family of proteases is comprised of the carboxypeptidase PRCP and the aminopeptidase DPP7. The structural basis of the different substrate specificities of the two enzymes is not understood nor has the structure of the S28 fold been described. RESULTS: The experimentally phased 2.8 A crystal structure is presented for human PRCP. PRCP contains an alpha/beta hydrolase domain harboring the catalytic Asp-His-Ser triad and a novel helical structural domain that caps the active site. Structural comparisons with prolylendopeptidase and DPP4 identify the S1 proline binding site of PRCP. A structure-based alignment with the previously undescribed structure of DPP7 illuminates the mechanism of orthogonal substrate specificity of PRCP and DPP7. PRCP has an extended active-site cleft that can accommodate proline substrates with multiple N-terminal residues. In contrast, the substrate binding groove of DPP7 is occluded by a short amino-acid insertion unique to DPP7 that creates a truncated active site selective for dipeptidyl proteolysis of N-terminal substrates. CONCLUSION: The results define the structure of the S28 family of proteases, provide the structural basis of PRCP and DPP7 substrate specificity and enable the rational design of selective PRCP modulators. Title: Discovery of conformationally rigid 3-azabicyclo[3.1.0]hexane-derived dipeptidyl peptidase-IV inhibitors Sattigeri JA, Andappan MM, Kishore K, Thangathirupathy S, Sundaram S, Singh S, Sharma S, Davis JA, Chugh A, Bansal VS Ref: Bioorganic & Medicinal Chemistry Lett, 18:4087, 2008 : PubMed ESTHER: Sattigeri_2008_Bioorg.Med.Chem.Lett_18_4087 PubMedSearch: Sattigeri 2008 Bioorg.Med.Chem.Lett 18 4087 PubMedID: 18602260 Abstract The induction of conformationally restricted N-(aryl or heteroaryl)-3-azabicyclo[3.1.0]hexane derivatives at P(2) region of compounds of 2-cyanopyrrolidine class was explored to develop novel DPP-IV inhibitors. The synthesis, structure-activity relationship, and selectivity against related proteases are delineated. Title: Acute toxicity bioassays of mercuric chloride and malathion on air-breathing fish Channa punctatus (Bloch) Pandey S, Kumar R, Sharma S, Nagpure NS, Srivastava SK, Verma MS Ref: Ecotoxicology & Environmental Safety, 61:114, 2005 : PubMed ESTHER: Pandey_2005_Ecotoxicol.Environ.Saf_61_114 PubMedSearch: Pandey 2005 Ecotoxicol.Environ.Saf 61 114 PubMedID: 15814317 Abstract Acute toxicity tests (96 h) were conducted in flow-through systems to determine the lethal toxicity of a heavy metal compound, mercuric chloride, and an organophosphorus pesticide, malathion, to air-breathing teleost fish, Channa punctatus (Bloch) and to study their behavior. The 96-h LC50 values were determined, as well as safe levels. The results indicate that mercuric chloride is more toxic than malathion to the fish species under study. Dose- and dose-time-dependent increases in mortality rate were also observed in response to both test chemicals. Title: Identification of a hypothetical membrane protein interactor of ribosomal phosphoprotein P0 Aruna K, Chakraborty T, Nambeesan S, Mannan AB, Sehgal A, Bhalchandara SR, Sharma S Ref: J Biosci, 29:33, 2004 : PubMed ESTHER: Aruna_2004_J.Biosci_29_33 PubMedSearch: Aruna 2004 J.Biosci 29 33 PubMedID: 15286401 Gene_locus related to this paper: yeast-yfd4 Abstract The ribosomal phosphoprotein P0 of the human malarial parasite Plasmodium falciparum (PfP0) has been identified as a protective surface protein. In Drosophila, P0 protein functions in the nucleus. The ribosomal function of P0 is mediated at the stalk of the large ribosomal subunit at the GTPase centre, where the elongation factor eEF2 binds. The multiple roles of the P0 protein presumably occur through interactions with other proteins. To identify such interacting protein domains, a yeast two-hybrid screen was carried out. Out of a set of sixty clones isolated, twelve clones that interacted strongly with both PfP0 and the Saccharomyces cerevisiae P0 (ScP0) protein were analysed. These belonged to three broad classes: namely (i) ribosomal proteins; (ii) proteins involved in nucleotide binding; and (iii) hypothetical integral membrane proteins. One of the strongest interactors (clone 67B) mapped to the gene YFL034W which codes for a hypothetical integral membrane protein, and is conserved amongst several eukaryotic organisms. The insert of clone 67B was expressed as a recombinant protein, and immunoprecipitaion (IP) reaction with anti-P0 antibodies pulled down this protein along with PfP0 as well as ScP0 protein. Using deletion constructions, the domain of ScP0, which interacted with clone 67B, was mapped to 60-148 amino acids. It is envisaged that the surface localization of P0 protein may be mediated through interactions with putative YFL034W-like proteins in P. falciparum. Title: Irreversible and reversible pore formation by polymeric alkylpyridinium salts (poly-APS) from the sponge Reniera sarai McClelland D, Evans RM, Abidin I, Sharma S, Choudhry FZ, Jaspars M, Sepcic K, Scott RH Ref: British Journal of Pharmacology, 139:1399, 2003 : PubMed ESTHER: McClelland_2003_Br.J.Pharmacol_139_1399 PubMedSearch: McClelland 2003 Br.J.Pharmacol 139 1399 PubMedID: 12922926 Inhibitor(s) related to this paper: 3-Alkylpyridinium-Polymers Abstract 1. In this study, we investigated the electrophysiological actions of a high molecular weight fraction, predominantly containing two polymeric 1,3-alkylpyridinium salts (poly-APS) of 5.5 and approximately 19 kDa isolated from the marine sponge Reniera sarai. The biological properties of poly-APS are of particular interest because this preparation may be used to deliver macromolecules into the intracellular environment without producing long-term damage to cells. Poly-APS (50-0.05 micro g ml(-1)) was applied to cultured dorsal root ganglion neurones or HEK 293 cells and changes in cell membrane properties were measured using whole-cell patch-clamp recording and fura-2 Ca(2+) imaging. 2. Poly-APS (50 micro g ml(-1)) evoked irreversible depolarisations in membrane potential and reductions in input resistance. However, doses of 5 micro g ml(-1) and less produced reversible effects on these cell membrane characteristics and on Ca(2+) permeability. 3. At 0.05 micro g ml(-1), poly-APS could robust transient increases in Ca(2+) permeability without damaging the neurones or subsequently attenuating Ca(2+) entry through voltage-activated channels. 4. Bathing cells in NaCl-based extracellular medium containing 1.5 mM zinc attenuated the irreversible and reversible effects of poly-APS on membrane properties (membrane potential, input resistance and whole-cell currents). In both DRG neurones and HEK 293 cells, zinc attenuated Ca(2+) entry evoked by poly-APS. These effects of zinc were only observed if zinc was continually present during poly-APS application. However, zinc failed to attenuate the actions of poly-APS if it was applied after the sponge toxin preparation had evoked changes in membrane properties. 5. In conclusion, the pore-forming preparation poly-APS can have dose-dependent interactions with cell membranes and at low doses these can be reversible. Additionally, the interactions between poly-APS and cell membranes could be attenuated by zinc. Title: Identification of novel bovine RPE and retinal genes by subtractive hybridization Sharma S, Chang JT, Della NG, Campochiaro PA, Zack DJ Ref: Mol Vis, 8:251, 2002 : PubMed ESTHER: Sharma_2002_Mol.Vis_8_251 PubMedSearch: Sharma 2002 Mol.Vis 8 251 PubMedID: 12131876 Gene_locus related to this paper: bovin-SPG21 Abstract PURPOSE: Understanding of the specialized function of the retinal pigment epithelium (RPE) can be aided by the identification and characterization of genes that are preferentially expressed in the RPE. With this aim, we undertook a systematic effort to identify and begin characterization of such genes. METHODS: A subtracted bovine RPE cDNA library was generated through subtractive hybridization using a single-stranded circular bovine RPE cDNA library as target and biotinylated mRNA from bovine heart and liver as alternate drivers. Approximately one thousand of the resulting subtracted cDNA clones were partially sequenced and analyzed, and a non-redundant set of one hundred of these cDNAs were examined for tissue expression pattern using a mini-Northern blot procedure and for identity by sequence analysis. RESULTS: The subtraction method successfully allowed the enrichment of cDNAs that are preferentially expressed in the RPE. Out of the analyzed clones, expression of forty-five clones was verifiable by Northern blotting. Of these, a significant proportion of cDNAs were preferentially expressed in the RPE. We observed that the expression of some subtracted cDNAs was restricted to the retina and no expression was detected in the RPE. These retinal clones were obtained in addition to RPE clones presumably because the initial RPE RNA population was contaminated with a small proportion of retinal RNA. Two thirds of the identified RPE and retinal cDNAs are likely to represent novel genes because they do not have homology to known genes in the databases. CONCLUSIONS: Genes that are specifically or predominantly expressed in the RPE/retina are likely to be important for retinal function. We have identified novel cDNAs from bovine RPE and retina by subtractive hybridization. These cDNAs can be used as starting material for the identification of corresponding human genes expressed in the RPE and retina. The human genes thus identified are likely to contain good candidate genes for retinal disease. Title: Oculohypotensive effect of angiotensin-converting enzyme inhibitors in acute and chronic models of glaucoma Shah GB, Sharma S, Mehta AA, Goyal RK Ref: J Cardiovasc Pharmacol, 36:169, 2000 : PubMed ESTHER: Shah_2000_J.Cardiovasc.Pharmacol_36_169 PubMedSearch: Shah 2000 J.Cardiovasc.Pharmacol 36 169 PubMedID: 10942157 Abstract We have studied the effects of various angiotensin-converting enzyme (ACE) inhibitors on intraocular pressure (IOP) of rabbits with experimentally induced ocular hypertension and their mechanism of action. Acute ocular hypertension was induced by infusion of 5% glucose (15 ml/kg) through marginal ear vein, whereas chronic glaucoma was induced by injection of alpha-chymotrypsin into the posterior chamber of the eye. IOP was measured by tonometer. All ACE inhibitors were instilled topically in the eye in a sterile solution. The effect of ACE inhibitors also was studied on serum cholinesterase (true and pseudo) and the enzyme ACE in vitro. Enalaprilat, ramiprilat, and fosinopril produced a time-dependent decrease of IOP in both acute and chronic models of ocular hypertension in rabbits. The decrease in IOP was observed for >4 h, and the extent of decrease was comparable to that with both pilocarpine and betaxolol. Prodrugs enalapril and ramipril failed to produced any change in IOP. Losartan also produced a significant decrease in IOP in the chronic model of ocular hypertension in rabbits. All the three ACE inhibitors were found to inhibit ACE activity in aqueous humor. The enzyme cholinesterase was found to be inhibited by enalaprilat, ramiprilat, and fosinopril. However, atropine did not alter the IOP-lowering effect of enalaprilat in rabbits. Indomethacin pretreatment produced slight but significant inhibition of the IOP-lowering effect of enalaprilat in rabbits. Our data suggest that ACE inhibitors enalaprilat, ramiprilat, and fosinopril produce a significant ocular hypotensive effect in acute and chronic models of ocular hypertension in rabbits. Inhibition of ACE in aqueous humor, and in ocular tissues, resulting in reduced angiotensin II formation, could be one of the major mechanisms responsible for the IOP reduction by ACE inhibitors in rabbits. Title: A spectrophotometric method for assay of tannase using rhodanine Sharma S, Bhat TK, Dawra RK Ref: Analytical Biochemistry, 279:85, 2000 : PubMed ESTHER: Sharma_2000_Anal.Biochem_279_85 PubMedSearch: Sharma 2000 Anal.Biochem 279 85 PubMedID: 10683234 Abstract A method for assay of microbial tannase (tannin acyl hydrolase) based on the formation of chromogen between gallic acid and rhodanine is reported. Unlike the previous protocols, this method is sensitive up to gallic acid concentration of 5 nmol and has a precision of 1.7% (relative standard deviation). The assay is complete in a short time, very convenient, and reproducible. Title: Characterization of secretory acetylcholinesterase from Setaria cervi microfilariae: a potential antigen for diagnosis of human filariasis Sharma S, Rathaur S Ref: Trop Med Int Health, 4:341, 1999 : PubMed ESTHER: Sharma_1999_Trop.Med.Int.Health_4_341 PubMedSearch: Sharma 1999 Trop.Med.Int.Health 4 341 PubMedID: 10402969 Abstract Acetylcholinesterase (AChE) is released to the external medium when microfilariae (m.f.) of Setaria cervi, a bovine filarial parasite, are maintained in vitro. Intense enzyme staining at amphids, excretory pores, anal vesicle and phasmids suggest an active secretion of AChE from m.f. Excretory-secretory products of m.f. displayed two electromorphic variants of AChE when resolved by 6% nondenaturing PAGE. The two isoforms of AChE (A and B) were separated on the basis of charge by DEAE sepharose CL 6B column following gel filtration. The two isoforms showed differing kinetic properties with respect to substrate specificity and inhibitor sensitivity. Anti-Nippostrongylus brasiliensis AChE antibodies cross-reacted with the affinity purified secretory AChE in ELISA. Immunoblotting of purified AChEs with cross-reacting anti-AChE antibodies revealed the presence of an approximately 75 kD protein in the isoenzyme A and an approximately 45 kD protein in B, whereas both proteins were present in the enzyme purified via affinity chromatography on edrophonium sepharose column. Title: Secretory acetylcholinesterase of Setaria cervi microfilariae and its antigenic cross-reactivity with Wuchereria bancrofti Sharma S, Misra S, Rathaur S Ref: Trop Med Int Health, 3:46, 1998 : PubMed ESTHER: Sharma_1998_Trop.Med.Int.Health_3_46 PubMedSearch: Sharma 1998 Trop.Med.Int.Health 3 46 PubMedID: 9484968 Abstract Setaria cervi, a bovine filarial parasite, secretes acetylcholinesterase during in vitro cultivation. A significant amount of enzyme activity was detected both in culture media and somatic extracts of different developmental stages of the parasite. The microfilarial stage showed a higher level of AChE activity than adult worms, with females being considerably more active than males. The secretory enzyme from microfilariae preferentially utilized acetylthiocholine iodide as substrate and showed two electrophoretically distinct isoforms in native PAGE. Secretory enzyme was purified from the excretory/secretory products of microfilariae using edrophonium chloride linked to epoxy-activated sepharose. Analysis of purified acetylcholinesterase by SDS-PAGE revealed the existence of two proteins of 75kD and 45kD under nonreducing conditions. These secretory enzymes are antigenic and cross-reactive with Wuchereria bancrofti-infected asymptomatic microfilaraemic human sera when tested by enzyme linked immunosorbent assay and immunoblotting. The secretory AChE(s) from S. cervi microfilariae may be utilized for diagnosis of early filarial infections. Title: Changes in cholinesterase activity in the genital tract, effect of olfactory lobe lesions Sharma S, Savithramma M, Sharma KN Ref: Indian Journal of Physiology & Pharmacology, 14:29, 1970 : PubMed ESTHER: Sharma_1970_Indian.J.Physiol.Pharmacol_14_29 PubMedSearch: Sharma 1970 Indian.J.Physiol.Pharmacol 14 29 | |||||||
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