Exploration of Potential Anti-inflammatory cum Anti-Rheumatoid-arthritis Phyto-molecule through Integrated Green Approach: Network Pharmacology, Molecular Docking, Molecular dynamics, In-vitro and Ex-vivo study.

Rheumatoid arthritis (RA) and associated inflammatory complications are the most prevalent illnesses and can turn into fatal conditions if left untreated. Allopathic medicine is not satisfactory for curing RA. Scientific literature reports reveal that several phyto-compounds viz. flavonoids, saponins, and terpenoids, can heal joints and organs from auto-inflammatory rheumatoid arthritis and pain. Gene ontology, gene network analysis, molecular clustering, and literature review were used to optimise RA-specific highly expressed genes. In-silico molecular docking was performed to short-out potential phytomolecules (Neohesperidin dihydrochalcone (NHDC)) from 1000 datasets-library against RA and validate using MD simulation running at 100 ns. In-vitro anti-inflammatory assays of NHDC inhibited egg-albumin denaturation, IC50 of 47.739 ± 0.51 µg/ml. The ex-vivo MTT assay with NHDC rendered 67.209% inhibition at 100 µM against fd-FLS-cells. NHDC downregulated pro-inflammatory cytokine IL-17A production by 61.11% and 50% at 300 and 200 µM, respectively. Thus, this Studies recommend that NHDC may be highlighted as a novel multi-target PADI4 and JAK3 inhibitor with better efficacy and minimal toxicity in RA warranted to In-Vivo and clinical investigation. The current findings have uncovered remarkable genes and signalling pathways linked to RA, which could enhance our existing comprehension of the molecular mechanisms that drive its development and progression.

Identifying druggable targets from active constituents of Azadirachta indica A. Juss. for non-small cell lung cancer using network pharmacology and validation through molecular docking.

INTRODUCTION: Azadirachta indica A. Juss. is a well-known medicinal plant that has been used traditionally to cure various ailments in every corner of the globe. There are many in vitro and in vivo experimental evidences in connection with the bioactivity of the extracts of this plant. Lung cancer is the deadliest form of cancer and contributes to the most cancer related deaths. The mode of action of anticancer components of this plant is still to be established explicitly. OBJECTIVE: The objective of this study is to identify druggable targets of active constituents of A. indica A. Juss. for non-small cell lung cancer (NSCLC) using network pharmacology and validation of activity through molecular docking analysis. METHODOLOGY: Targets of all the active phytochemicals from A. indica were predicted and genes related to NSCLC were retrieved. A protein-protein interaction (PPI) network of the overlapping genes were prepared. Various databases and servers were employed to analyse the disease pathway enrichment analysis of the clustered genes. Validation of the gene/protein activity was achieved by performing molecular docking, and ADMET profiling of selected phytocompounds was performed. RESULT: Gene networking revealed three key target genes as EGFR, BRAF and PIK3CA against NSCLC by the active components of A. indica. Molecular docking and ADMET analysis further validated that desacetylnimbin, nimbandiol, nimbin, nimbinene, nimbolide, salannin and vepinin are the best suited anti- NSCLC among all the phytocompounds present in this plant. CONCLUSION: The present study has provided a better understanding of the pharmacological effects of active components from A. indica and its potential therapeutic effect on NSCLC.

Transcriptional expression of secondary resistance genes ccdB and repA2 is enhanced in presence of cephalosporin and carbapenem in Escherichia coli.

BACKGROUND: The issue of carbapenem resistance in E.coli is very concerning and it is speculated that cumulative effect of both primary resistance genes and secondary resistance genes that act as helper to the primary resistance genes are the reason behind their aggravation. Therefore, here we attempted to find the role of two secondary resistance genes (SRG) ccdB and repA2 in carbapenem resistance in E. coli (CRE). In this context influential genes belonging to secondary resistome that act as helper to the primary resistance genes like blaNDM and blaCTX-M in aggravating ß-lactam resistance were selected from an earlier reported in silico study. Transcriptional expression of the selected genes in clinical isolates of E.coli that were discretely harboring blaNDM-1, blaNDM-4, blaNDM-5, blaNDM-7 and blaCTX-M-15 with and without carbapenem and cephalosporin stress (2 µg/ml) was determined by real time PCR. Cured mutants sets that were lacking (i) primary resistance genes, (ii) secondary resistance genes and (iii) both primary and secondary resistance genes were prepared by SDS treatment. These sets were then subjected to antibiotic susceptibility testing by Kirby Bauer disc diffusion method. RESULTS: Out of the 21 genes reported in the in silico study, 2 genes viz. repA2 and ccdB were selected for transcriptional expression analysis. repA2, coding replication regulatory protein, was downregulated in response to carbapenems and cephalosporins. ccdB, coding for plasmid maintenance protein, was also downregulated in response to carbapenems except imipenem and cephalosporins. Following plasmid elimination assay increase in diameter of zone of inhibition under stress of both antibiotics was observed as compared to uncured control hinting at the reversion of antibiotic susceptibility by the-then resistant bacteria. CONCLUSION: SRGs repA2 and ccdB help sustenance of blaNDM and blaCTX-M under carbapenem and cephalosporin stress.

Sublethal effects of pulp and paper mill effluent on two commonly cultured carps: a SEM- and EDS-based hematological biomarker analysis.

Blood being a vehicle for the transport of industrial pollutants in living system, fish hematology is considered as potent biomarker. In the present study, we investigated respective sublethal effects of pulp and paper mill effluents on hematology of two commonly cultured carps, Cyprinus carpio and Ctenopharyngodon idella, using optical, scanning electron microscopy and energy-dispersive X-ray spectroscopy. Irrespective of species, results showed significant decrease in erythrocyte, hematocrit and hemoglobin contents while an increase in white blood cell counts (P < 0.05). We observed an increasing trend of MCV (170.0 ± 3.07 to 193.16 ± 2.5) and MCH (34.31 ± 1.89 to 38.71 ± 3.61) up to 28th day in C. carpio (P < 0.05), while, in C. idella, the highest percent increase in MCV (180.8 ± 2.19) and MCH (32.9 ± 0.62) was observed on seventh exposure day, which subsequently declined, respectively, to 173.1 ± 17.1 and 27.9 ± 2.45 on 28th day. Unlike C. carpio, significant and progressive MCHC declining trend (18.23 ± 0.28 to 16.13 ± 0.31) was observed in C. idella. The most commonly observed abnormalities under SEM include echinocytes, cytoplasmic blebbing, cytoplasmic ring, spherocytes, lobopodial projections and acanthocytes in red blood cells of exposed fishes. EDS further revealed the presence of aluminum, antimony, arsenic, cadmium, mercury, tungsten, zinc and titanium; some of these metals were not even detected in the effluent samples, suggesting the probable metal bio-concentration in fish tissue, and subsequent jeopardization is a major concern particularly in the industrial area. Our study further suggested the use of sensitive and specific techniques like SEM and EDS in fish hematological biomarker analysis along with the conventional approach.

NoSQL data model for semi-automatic integration of ethnomedicinal plant data from multiple sources.

INTRODUCTION: Sharing traditional knowledge with the scientific community could refine scientific approaches to phytochemical investigation and conservation of ethnomedicinal plants. As such, integration of traditional knowledge with scientific data using a single platform for sharing is greatly needed. However, ethnomedicinal data are available in heterogeneous formats, which depend on cultural aspects, survey methodology and focus of the study. Phytochemical and bioassay data are also available from many open sources in various standards and customised formats. OBJECTIVE: To design a flexible data model that could integrate both primary and curated ethnomedicinal plant data from multiple sources. MATERIALS AND METHODS: The current model is based on MongoDB, one of the Not only Structured Query Language (NoSQL) databases. Although it does not contain schema, modifications were made so that the model could incorporate both standard and customised ethnomedicinal plant data format from different sources. RESULTS: The model presented can integrate both primary and secondary data related to ethnomedicinal plants. Accommodation of disparate data was accomplished by a feature of this database that supported a different set of fields for each document. It also allowed storage of similar data having different properties. CONCLUSION: The model presented is scalable to a highly complex level with continuing maturation of the database, and is applicable for storing, retrieving and sharing ethnomedicinal plant data. It can also serve as a flexible alternative to a relational and normalised database.

Validation of novel catechol derivatives as potent Escherichia coli MetAP inhibitors using Molecular Docking and Molecular Dynamics Simulation.

BACKGROUND: Urinary tract infections (UTIs) are the most common form of nosocomial infection primarily caused by Escherichia coli. Complicated UTIs carry a higher risk of treatment failure, recurrent infections, and increased morbidity. Methionine aminopeptidase (MetAP) has gained tremendous importance as a bacterial drug target due to its role in cell growth and membrane integrity. However, the participation of metal-chelating residues and the occurrence of the enzyme in the human body complicate the process of selecting a suitable inhibitor. AIMS: This study aimed to find new molecules with more stable binding against urinary tract infection drug targets. OBJECTIVE: The objective of this study was to find new molecules with more stable binding against urinary tract infection drug targets using computational approaches. METHOD: The drug target was selected based on a literature study. Catechol derivatives were prepared and an ADME/T study was performed, followed by molecular docking and molecular dynamics. RESULT: The docking score of Met592 (-20.95) was found to be much better than that of known inhibitors (-12.88). The overall study on Rg signified that the ligand binding compels the respective proteins to become more compact and less flexible in the case of Met592. Binding free energy analysis also showed a better affinity for Met592 (-46.60) than the known inhibitor (-31.37). CONCLUSION: The increased binding score, good oral bioavailability, and better binding free energy endorse the reliability of the ligand Met592, i.e., (R, E)-4-(4-(2-(((9H-purin-6-yl)amino)methyl)- 4,5-dimethylphenyl)thiazol-2-yl)-4-aminobut-2-enoic acid, as the probable drug candidate to treat uropathogenic E. coli.

Ethnomedicinal, Phytochemical and Nutra-pharmaceutical Potentials of Indian Arrowroot (Curcuma angustifolia Roxb).

Indian Arrowroot (Curcuma angustifolia Roxb) belonging to the Zingiberaceae family is widely distributed in India and some parts of Nepal, Thailand, Bangladesh and Pakistan. It is traditionally used as medicine for treating various diseases and also used as food. Few data are available about its application in pharmacology and therapeutics. Literature search for related contents, keywords such as "Curcuma angustifolia Roxb", "traditional food", "ethnomedicine", "pharmacology", "phytochemicals", "pharmacological activities" were used in search engines including PubMed, Google Scholar, Scopus, ScienceDirect, and Semantic Scholar. Secondary metabolites found in Indian Arrowroot include essential oils, alkaloids, flavonoids, terpenoids, phytosterols, terpenes, phenols, and others. Pharmacological activities such as antioxidant, antiinflammatory, anti-proliferative, anti-ulcerogenic, hepatoprotective, and anti-cancerous activities have been shown by Indian Arrowroot (Curcuma angustifolia Roxb). The presence of nutritional value and pharmaceutical potential gained demand in the various food production industries and pharmacology research. It may play a vital role in future studies of Curcuma angustifolia Roxb as ethnomedicine and further exploitation in pharmacological studies.

Natural Flavonoids in the Prevention and Treatment of Lung Cancer: A Pharmacological Aspect.

Deadly disease cancer has many types; among them, lung cancer is responsible for the highest number of cancer mortality. Existing therapies as well as drugs for treating lung cancer are not effective and are often associated with innumerable side effects and toxicities. For these reasons, researchers have been working on developing novel anti-cancer medicines from plants and other natural sources that have a high safety profile. Natural flavonoids are a polyphenolic group of phytochemicals extracted from plants and other plant-derived compounds. Natural flavonoids are gaining popularity due to their unique and priceless medicinal properties, including anticancer properties. Several researchers have already declared that flavonoids possess the ability to treat different cancers, particularly lung cancer. The bioactivity of natural flavonoids is mainly due to their structural diversity. Natural flavonoids fight against lung cancer by regulating redox homeostasis, upregulating apoptosis, pro-apoptotic factors, and survival genes, arresting cell cycle progression, autophagy, reducing cell proliferation and invasiveness, maintaining inflammation response, downregulating anti-apoptotic factors, and targeting lung cancer signaling pathways. Flavonoids can act alone or synergistically with other agents to treat lung cancer. Due to these reasons, it is possible to use natural flavonoids as pharmaceutical leads to prevent and treat lung cancer.

Effect of acetone fraction of Ottelia alismoides on the G2/M cell cycle arrest and apoptosis in the human carcinoma cell lines.

ETHNOPHARMACOLOGICAL RELEVANCE: The North-eastern parts of India have immense therapeutic floras, Ottelia alismoides is an aquatic plant that has been in use for a long time in traditional medicine for treating diseases like cancer, tuberculosis, diabetes, febrifuge, hemorrhoids, and rubefacient. In lung and skin carcinoma cells with a high rate of proliferation and metastasis including drug resistance and non-specific target activity, generates important challenges towards their treatment strategy. Thus, finding novel therapeutic targets to treat lung and skin cancer progression is essential to enhance the patients' survival with treatment. AIM OF THE STUDY: The purpose of this study was to evaluate the apoptotic potential of acetone extract of O. alismoides (L.) Pers. (OA-AC) and to identify the compounds responsible for this effect, HRLC-MS-QTOF analysis of the extract has been undertaken along with in-silico molecular docking analysis of the identified compounds. MATERIALS AND METHODS: A549 and A431 cells were treated with acetone extract of O. alismoides (OA-AC) at 24 h and 48 h exposure and cell cycle phase distribution was evaluated and also apoptosis induction activity was evaluated by OA-EtBr staining and Mitochondrial outer membrane potential assay. Western blotting was performed for the evaluation of apoptotic protein expression. At last, the HR-LCMS of OA-AC was analyzed to identify the compounds responsible for the apoptotic activity of the extract. RESULTS: The cell cycle phase distribution analysis in A549 and A431 cells at 24hrs exposure with 10 µg/mL and 25 µg/mL of OA-AC showed a potent arrest or blockage at the G2/M phase of the cell cycle with reduced expression of cyclin B and p-Cdc2. At 48 h exposure, apoptosis was observed in these cancer cells with elevated expression of Bax, p21 and cleaved caspase 3 and reduced expression of the Bcl2. CONCLUSION: AO-EtBr staining of these cancer cells reveals that the death induced by OA-AC was apoptotic in nature with depolarization of mitochondrial membrane due to loss or damage of the mitochondrial membrane. The HRLC-MS-QTOF analysis of OA-AC depicted 14 major isolable compounds and molecular docking analysis displayed 4 compounds that might act as an inhibitor of cyclin B for G2/M phase arrest that leads to apoptotic induction in the cells.

Network analysis of atherosclerotic genes elucidates druggable targets.

BACKGROUND: Atherosclerosis is one of the major causes of cardiovascular disease. It is characterized by the accumulation of atherosclerotic plaque in arteries under the influence of inflammatory responses, proliferation of smooth muscle cell, accumulation of modified low density lipoprotein. The pathophysiology of atherosclerosis involves the interplay of a number of genes and metabolic pathways. In traditional translation method, only a limited number of genes and pathways can be studied at once. However, the new paradigm of network medicine can be explored to study the interaction of a large array of genes and their functional partners and their connections with the concerned disease pathogenesis. Thus, in our study we employed a branch of network medicine, gene network analysis as a tool to identify the most crucial genes and the miRNAs that regulate these genes at the post transcriptional level responsible for pathogenesis of atherosclerosis. RESULT: From NCBI database 988 atherosclerotic genes were retrieved. The protein-protein interaction using STRING database resulted in 22,693 PPI interactions among 872 nodes (genes) at different confidence score. The cluster analysis of the 872 genes using MCODE, a plug-in of Cytoscape software revealed a total of 18 clusters, the topological parameter and gene ontology analysis facilitated in the selection of four influential genes viz., AGT, LPL, ITGB2, IRS1 from cluster 3. Further, the miRNAs (miR-26, miR-27, and miR-29 families) targeting these genes were obtained by employing MIENTURNET webtool. CONCLUSION: Gene network analysis assisted in filtering out the 4 probable influential genes and 3 miRNA families in the pathogenesis of atherosclerosis. These genes, miRNAs can be targeted to restrict the occurrence of atherosclerosis. Given the importance of atherosclerosis, any approach in the understanding the genes involved in its pathogenesis can substantially enhance the health care system.

Colistin exposure enhances expression of eptB in colistin-resistant Escherichia coli co-harboring mcr-1.

Colistin resistance has increased due to the increasing and inappropriate use of this antibiotic. The mechanism involves modification of lipid A with phosphoethanolamine (PEtN) and/or 4-amino-4deoxy-L-arabinose (L-Ara4N). EptA and eptB catalyze the transfer of phosphoethanolamine to lipid A. In this study, gene network was constructed to find the associated genes related to colistin resistance, and further in vitro validation by transcriptional analysis was performed. In silico studies showed that eptB gene is a highly interconnected node in colistin resistance gene network. To ascertain these findings twelve colistin-resistant clinical isolates of Escherichia coli were selected in which five were harboring the plasmid-mediated mcr-1. Screening for colistin resistance was performed by broth microdilution (BMD) method and Rapid polymyxin NP test. PCR confirmed the presence of the eptA and eptB genes in all isolates and five isolates were harboring mcr-1. Transcriptional expression in five isolates harboring mcr-1, showed an enhanced expression of eptB when exposed under sub-inhibitory colistin stress. The present study for the first time highlighted genetic interplay between mcr-1 and eptA and eptB under colistin exposure.

The chimera of S1 and N proteins of SARS-CoV-2: can it be a potential vaccine candidate for COVID-19?

INTRODUCTION: Coronavirus disease 2019 (COVID-19), caused by the severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), has emerged as one of the biggest global health issues. Spike protein (S) and nucleoprotein (N), the major immunogenic components of SARS-CoV-2, have been shown to be involved in the attachment and replication of the virus inside the host cell. AREAS COVERED: Several investigations have shown that the SARS-CoV-2 nucleoprotein can elicit a cell-mediated immune response capable of regulating viral replication and lowering viral burden. However, the development of an effective vaccine that can stop the transmission of SARS-CoV-2 remains a matter of concern. Literature was retrieved using the keywords COVID-19 vaccine, role of nucleoprotein as vaccine candidate, spike protein, nucleoprotein immune responses against SARS-CoV-2, and chimera vaccine in PubMed, Google Scholar, and Google. EXPERT OPINION: We have focussed on the use of chimera protein, consisting of N and S-1 protein components of SARS-CoV-2, as a potential vaccine candidate. This may act as a polyvalent mixed recombinant protein vaccine to elicit a strong T and B cell immune response, which will be capable of neutralizing the wild and mutated variants of SARS-CoV-2, and also restricting its attachment, replication, and budding in the host cell.

Cadmium toxicity at low concentration on rabbit spermatozoa motility, morphology and membrane integrity in vitro.

In this study the effect of cadmium on various parameters of spermatozoa motility, morphology as well as on the spermatozoa membrane integrity in rabbits was analyzed in vitro, experimental concentrations ranging from 0.62 to 0.98 micro g CdCl(2)/mL. Pooled rabbit (n = 5) semen was cultured in vitro with cadmium and subsequently diluted to various experimental concentrations apart from control which received no cadmium exposure. Using computer assisted semen analysis method (CASA) we detected decrease of total motility with in the higher concentration range at Time 0. However, with increasing time (after 1 and 2 h of culture), cadmium exerted deleterious effect leading to significant motility reduction in comparison to control. A similar trend was exhibited in case of progressive motility, too. Most of the spermatozoa distance and velocity parameters detected no significant change in comparison to control at the beginning of culture (Time 0), although the toxic effect became significant (P < 0.05) with the passage of culture time (Times 1 and 2 h) in all concentrations. Analysis of spermatozoa morphology detected significant (P < 0.05) alterations at higher concentrations. At higher concentrations acrosomal changes, head without flagellum/separated flagellum, broken flagellum and other abnormalities were significantly higher (P < 0.05), while knob-twisted flagellum and small heads differed significantly (P < 0.05) in comparison to control at all concentrations. In regards to flagellum torso, flagellum ball and retention of cytoplasmic drop statistically higher values (P < 0.05) were noted at the maxium experimental concentration only. Annexin analysis for detection of spermatozoa with disordered membranes revealed higher occurrence of positive spermatozoa in cadmium exposed groups. Annexin-positive reactions suggested alterations in anterior part of head (acrosome) and in flagellum (mitochondrial segment) of spermatozoa. This paper underlines that cadmium is highly toxic for rabbit spermatozoa, as visualized by the toxic effects on parameters of spermatozoa motility, morphology and membrane integrity. The toxic effect is more drastic at higher concentrations. This study also indicates that cadmium requires a minimum one hour incubation time to exert its deletorious effects on various parameters of spermatozoa, particularly at low concentrations.

In vitro copper toxicity on rabbit spermatozoa motility, morphology and cell membrane integrity.

In this in vitro study the effects of copper sulphate on the motility, morphology and structural integrity of rabbit spermatozoa were investigated. The spermatozoa motility was evaluated by CASA method and Annexin analysis was used for detection of structural changes. For analysis of morphology samples of rabbit semen were fixed with Hancock's solution and stained with Giemsa, and for each sample at least 500 spermatozoa were evaluated. The concentration of copper in the medium varied from 3.57 to 4.85 microg CuSO4/mL. At Time 0 the highest motility was detected in the control group (57.78 +/- 3.90%). Motility in groups with copper administration was lower in comparison to control. Significant differences were detected in groups with 3.70-4.85 microg CuSO4/mL (P<0.05) at Time 0. After 1 h of incubation with copper sulphate the motility significantly decreased almost in all experimental groups. However, at Time 2 h significant increase of total motility was observed in groups with lower concentrations of copper (3.57 and 3.63 microg CuSO4/mL). After 24 and 48 h of incubation almost all the spermatozoa were dead recording no motility at all concentrations. The concentration- dependent decrease of spermatozoa motility up to 50% of control was detected for the group receiving highest copper administration (4.85 microg CuSO4/mL) at Times 1 and 2 h. Progressive motility had an identical trend to that of motility in all experimental groups, at all culture times and for all concentrations. Evaluation of distance and velocity parameters indicated that a sort of stress tolerance developed in lower concentrations (3.57 and 3.63 microg CuSO4/mL). At lower concentrations, an increase was noted for distance parameter DCL and velocity parameter VCL, indirectly confirming the significant motility and progressive motility increase. Other motility parameters (straightness index, linearity index, wobble and amplitude of lateral head displacement) revealed decrease in the group with the highest copper concentration (4.85 microg CuSO4/mL) in comparison to the control group after 2 h of incubation, only. No significant alteration was noted for these parameters in comparison to control at Times 0 and 1 h. The total percentage of morphologically abnormal spermatozoa was significantly higher (P<0.05) in the group with the highest copper concentration (46.20+/-5.54%) in comparison to control (30.60+/-2.91). Predominant morphological abnormalities were acrosomal changes, knob-twisted flagellum and small heads. Detection of spermatozoa with disordered membrane was carried out for groups with higher copper concentrations and control, using Annexin analysis. Analysis showed higher occurrence of positive spermatozoa in the copper-exposed groups. Some Annexin positive reactions from all spermatozoa were detected in the control group. In copper-exposed groups positive reaction proved alteration in anterior part of head (acrosome) and in connection segment (mid-piece) of spermatozoa. Detected data evidently confirm adverse effects of high copper sulphate concentrations in rabbit semen on parameters of spermatozoa motility, morphology and membrane integrity. This paper also indicates the lowest possible toxic concentration of copper (3.70 microg CuSO4/mL) to rabbit spermatozoa in relation to motility.

Role of Natural Phenolics in Hepatoprotection: A Mechanistic Review and Analysis of Regulatory Network of Associated Genes.

The liver is not only involved in metabolism and detoxification, but also participate in innate immune function and thus exposed to frequent target Thus, they are the frequent target of physical injury. Interestingly, liver has the unique ability to regenerate and completely recoup from most acute, non-iterative situation. However, multiple conditions, including viral hepatitis, non-alcoholic fatty liver disease, long term alcohol abuse and chronic use of medications can cause persistent injury in which regenerative capacity eventually becomes dysfunctional resulting in hepatic scaring and cirrhosis. Despite the recent therapeutic advances and significant development of modern medicine, hepatic diseases remain a health problem worldwide. Thus, the search for the new therapeutic agents to treat liver disease is still in demand. Many synthetic drugs have been demonstrated to be strong radical scavengers, but they are also carcinogenic and cause liver damage. Present day various hepatic problems are encountered with number of synthetic and plant based drugs. Nexavar (sorafenib) is a chemotherapeutic medication used to treat advanced renal cell carcinoma associated with several side effects. There are a few effective varieties of herbal preparation like Liv-52, silymarin and Stronger neomin phages (SNMC) against hepatic complications. Plants are the huge repository of bioactive secondary metabolites viz; phenol, flavonoid, alkaloid etc. In this review we will try to present exclusive study on phenolics with its mode of action mitigating liver associated complications. And also its future prospects as new drug lead.

Carbapenem nonsusceptibility with modified OprD in clinical isolates of Pseudomonas aeruginosa from India.

This study was undertaken to investigate OprD porin-mediated carbapenem nonsusceptibility in clinical isolates of Pseudomonas aeruginosa from a tertiary referral hospital of Northeast India. A total of 267 nonduplicate, consecutive clinical isolates of P. aeruginosa were obtained. Mutation and expression levels of OprD gene were determined in carbapenem-nonsusceptible carbapenemase-nonproducing isolates. Among 19 carbapenem-nonsusceptible carbapenemase-nonproducing isolates, 11 of them demonstrated variable band pattern while performing denaturing gradient gel electrophoresis with amplified products of OprD gene. Sequencing of variable band products revealed three mutation patterns in three isolates. Relevant decrease in expression of OprD gene could also be observed in them. All the three isolates exhibited a higher minimum inhibitory concentration for imipenem (64-128 µg/mL) compared to meropenem (16-64 µg/mL). Inactivating mutation and decreased expression of OprD contribute mainly to imipenem resistance as well as to meropenem.

Hypercholesterolemia causes psychomotor abnormalities in mice and alterations in cortico-striatal biogenic amine neurotransmitters: Relevance to Parkinson's disease.

The symptoms of Parkinson's disease (PD) include motor behavioral abnormalities, which appear as a result of the extensive loss of the striatal biogenic amine, dopamine. Various endogenous molecules, including cholesterol, have been put forward as putative contributors in the pathogenesis of PD. Earlier reports have provided a strong link between the elevated level of plasma cholesterol (hypercholesterolemia) and onset of PD. However, the role of hypercholesterolemia on brain functions in terms of neurotransmitter metabolism and associated behavioral manifestations remain elusive. We tested in Swiss albino mice whether hypercholesterolemia induced by high-cholesterol diet would affect dopamine and serotonin metabolism in discrete brain regions that would precipitate in psychomotor behavioral manifestations. High-cholesterol diet for 12 weeks caused a significant increase in blood total cholesterol level, which validated the model as hypercholesterolemic. Tests for akinesia, catalepsy, swimming ability and gait pattern (increased stride length) have revealed that hypercholesterolemic mice develop motor behavioral abnormalities, which are similar to the behavioral phenotypes of PD. Moreover, hypercholesterolemia caused depressive-like behavior in mice, as indicated by the increased immobility time in the forced swim test. We found a significant depletion of dopamine in striatum and serotonin in cortex of hypercholesterolemic mice. The significant decrease in tyrosine hydroxylase immunoreactivity in striatum supports the observed depleted level dopamine in striatum, which is relevant to the pathophysiology of PD. In conclusion, hypercholesterolemia-induced depleted levels of cortical and striatal biogenic amines reported hereby are similar to the PD pathology, which might be associated with the observed psychomotor behavioral abnormalities.

Molecular and in silico analysis of a new plasmid-mediated AmpC ß-lactamase (CMH-2) in clinical isolates of Klebsiella pneumoniae.

Two Klebsiella strains isolated from urine samples were positive for blaAmpC by PCR and showed sequence similarity with CMH-1 (98.6%) after sequencing. It also shares 82% similarity with ACT-1, 85% with MIR-1 and 81% with the chromosomal AmpC gene of Enterobacter cloacae. This gene was associated with the plasmid of IncK type. It has an open reading frame of 381 amino acid with four amino acid substitutions at position D144A, C189R, Q192E, and A195T as compared to CMH-1. When expressed in E.coli DH5α and E.coli strain B, this ß-lactamase conferred resistance to cefotaxime, ceftriaxone and ceftazidime. In addition, both in vitro and in silico analysis revealed that this cephalosporinase was inhibited by cefepime and carbapenem group of drugs. Therefore, this new plasmid-encoded AmpC type ß-lactamase gene was designated as CMH-2.

Screening of Natural Products and Derivatives for the Identification of RND Efflux Pump Inhibitors.

AIM AND OBJECTIVE: Overexpression of efflux pumps belonging to the Resistance Nodulation cell Division (RND) family is the most important intrinsic resistance mechanism of Pseudomonas aeruginosa. Hence, it is imperative to identify suitable efflux pump inhibitors (EPI) that can lead to increased intracellular concentration of antibiotics by blocking the pump. This study was undertaken to identify a putative plant based efflux pump inhibitor for RND efflux pump of P. aeruginosa. MATERIAL AND METHOD: Using molecular docking approach, 328 secondary plant metabolites have been screened for their inhibitory activity against cytoplasmic exporter protein MexB of MexAB-OprM efflux pump of P. aeruginosa. After the initial in silico screening, the shortlisted compounds were subjected to in vitro test for efflux pump inhibitory activity using double disc synergy test. A combinatorial library of 1000 molecules was generated from active p-coumaric acid and docked with MexB protein to find a suitable EPI with better binding efficacy compared to the p-coumaric acid. RESULTS: Preliminary screening resulted in five plant-based natural products with significant docking score and were subsequently subjected to double disc synergy test. p-Coumaric acid , amongst the five, was found to potentiate activity of ciprofloxacin in MexAB-OprM overexpressing P. aeruginosa strain. Library compound 482, i.e 4-(4-((Z)-2-carboxy-2-((Z)-2,3-dihydrobenzo[e][1,4]diazepin-1-yl)-1-(4- hydroxyphenyl)vinylamino) phenylsulfonamido)-2-hydroxybenzoic acid, a derivative of p-coumaric acid exhibited the highest docking score of -42.1030 Kcal/mol, which was much higher than parent compound (-17.9403 Kcal/mol) and also known EPI, MC-207,110 (-28.0960 Kcal/mol). CONCLUSION: p-Coumaric acid and its derivative, 4-(4-((Z)-2-carboxy-2-((Z)-2,3-dihydrobenzo[e][1,4] diazepin-1-yl)-1-(4-hydroxyphenyl)vinylamino)phenylsulfonamido)-2-hydroxybenzoic acid may be used as potential lead molecules for effective RND efflux pump inhibition in P. aeruginosa.

Curcumin as potential therapeutic natural product: a nanobiotechnological perspective.

OBJECTIVES: Nanotechnology-based drug delivery systems can resolve the poor bioavailability issue allied with curcumin. The therapeutic potential of curcumin can be enhanced by making nanocomposite preparation of curcumin with metal oxide nanoparticles, poly lactic-co-glycolic acid (PLGA) nanoparticles and solid lipid nanoparticles that increases its bioavailability in the tissue. KEY FINDINGS: Curcumin has manifold therapeutic effects which include antidiabetic, antihypertensive, anticancer, anti-inflammatory and antimicrobial properties. Curcumin can inhibit diabetes, heavy metal and stress-induced hypertension with its antioxidant, chelating and inhibitory effects on the pathways that lead to hypertension. Curcumin is an anticancer agent that can prevent abnormal cell proliferation. Nanocurcumin is an improved form of curcumin with enhanced therapeutic properties due to improved delivery to the diseased tissue, better internalization and reduced systemic elimination. SUMMARY: Curcumin has multiple pharmacologic effects, but its poor bioavailability reduces its therapeutic effects. By conjugating curcumin to metal oxide nanoparticles or encapsulation in lipid nanoparticles, dendrimers, nanogels and polymeric nanoparticles, the water solubility and bioavailability of curcumin can be improved and thus increase its pharmacological effectiveness.