Identification of DprE1 inhibitors for tuberculosis through integrated in-silico approaches.

Decaprenylphosphoryl-ß-D-ribose-2'-epimerase (DprE1), a crucial enzyme in the process of arabinogalactan and lipoarabinomannan biosynthesis, has become the target of choice for anti-TB drug discovery in the recent past. The current study aims to find the potential DprE1 inhibitors through in-silico approaches. Here, we built the pharmacophore and 3D-QSAR model using the reported 40 azaindole derivatives of DprE1 inhibitors. The best pharmacophore hypothesis (ADRRR_1) was employed for the virtual screening of the chEMBL database. To identify prospective hits, molecules with good phase scores (> 2.000) were further evaluated by molecular docking studies for their ability to bind to the DprE1 enzyme (PDB: 4KW5). Based on their binding affinities (< - 9.0 kcal/mole), the best hits were subjected to the calculation of free-binding energies (Prime/MM-GBSA), pharmacokinetic, and druglikeness evaluations. The top 10 hits retrieved from these results were selected to predict their inhibitory activities via the developed 3D-QSAR model with a regression coefficient (R2) value of 0.9608 and predictive coefficient (Q2) value of 0.7313. The induced fit docking (IFD) studies and in-silico prediction of anti-TB sensitivity for these top 10 hits were also implemented. Molecular dynamics simulations (MDS) were performed for the top 5 hit molecules for 200 ns to check the stability of the hits with DprE1. Based on their conformational stability throughout the 200 ns simulation, hit 2 (chEMBL_SDF:357100) was identified as the best hit against DprE1 with an accepted safety profile. The MD results were also in accordance with the docking score, MM-GBSA value, and 3D-QSAR predicted activity. The hit 2 molecule, (N-(3-((2-(((1r,4r)-4-(dimethylamino)cyclohexyl)amino)-9-isopropyl-9H-purin-6-yl)amino)phenyl)acrylamide) could serve as a lead for the discovery of a novel DprE1 inhibiting anti-TB drug.

Structure-activity relationship mediated molecular insights of DprE1 inhibitors: A Comprehensive Review.

Emerging threats of multi-drug resistant (MDR), extensively drug-resistant (XDR), and totally drug-resistant (TDR) tuberculosis led to the discovery of a novel target which was entitled Decaprenylphosphoryl-ß-D-ribose 2'-epimerase (DprE1) enzyme. DprE1 is composed of two isoforms, decaprenylphosphoryl-ß-D-ribose oxidase (DprE1) and decaprenylphosphoryl-D-2-keto erythro pentose reductase (DprE2). The enzymes, DprE1 and DprE2, regulate the two-step epimerization process to form DPA (Decaprenylphosphoryl arabinose) from DPX (Decaprenylphosphoryl-D-ribose), which is the sole precursor in the cell wall synthesis of arabinogalactan (AG) and lipoarabinomannan (LAM). Target-based and whole-cell-based screening played an imperative role in the identification of the druggable target, DprE1, whereas the druggability of the DprE2 enzyme is not proved yet. To date, diverse scaffolds of heterocyclic and aromatic ring systems have been reported as DprE1 inhibitors based on their interaction mode, i.e. covalent, and non-covalent inhibitors. This review describes the structure-activity relationship (SAR) of reported covalent and non-covalent inhibitors to enlighten about the crucial pharmacophoric features required for DprE1 inhibition, along with in-silico studies which characterize the amino acid residues responsible for covalent and non-covalent interactions.Communicated by Ramaswamy H. Sarma.

Sonophotocatalytic disinfection of Shigella species under visible light irradiation: Insights into its molecular mechanism, antibacterial resistance and biofilm formation.

Microbial contamination of water is one of the major sources of many diseases worldwide. Evolution of antibacterial resistance (ABR) alongside the caveats in most of the water treatment methods causes the severity of the current problem extremely vexing. This calls for an urgent need to develop new treatment methods aiming to reduce the microbial as well as ABR load in the environment. Herein, we successfully developed a visible light assisted sonophotocatalysis (SPC) using Fe/ZnO nanoparticles (NPs) for the disinfection of Shigella dysenteriae. A consortia containing S. dysenteriae and S. flexineri was also completely disinfected using SPC. Growth conditions of S. dysenteriae like growth phases and growth temperaturehad different outcomes on the overall efficacy of SPC. Compared with catalysts such as ZnO and TiO2, Fe/ZnO resulted in better disinfection. Multi-ROS production, mostly containing h+ and O2· radicals, due to the electron displacement in the catalyst and acoustic cavitation was identified as the factors behind bacterial lethality. The ROS produced was found to interfere with the metabolic activities of S. dysenteriae by causing membrane perturbation. We identified DNA damage inside the cells and the subsequent release of intracellular components. The compositional changes in the fatty acid makeup of the cells were altered as a result of SPC and few fatty acid markers indicating the stress posed by SPC were also identified. Loss of ABR in S. dysenteriae was also recorded post SPC treatment. Abatement in the biofilm forming ability of the injured bacterial cells was also recorded, proving the extremity of stress induced by SPC. Hence, the excellent efficacy of SPC in disinfecting bacteria is proposed for tertiary water treatment applications.

Microbial pretreatment of lignocellulosic biomass for enhanced biomethanation and waste management.

Biogas obtained from organic remains entails a developed technology and an appreciable methane yield, but its use may not be sustainable. The potential methane yield of various lignocellulose biomass and the operational conditions employed are inherently reviewed. Although of lower methane yields compared to conventional substrates, agricultural biomass is a cheap option. The major challenges encountered during its biogasification are its recalcitrance nature primarily due to the presence of crystalline cellulose and lignin. This necessitates an essential pretreatment step through physical, chemical or biological interventions for enhanced biomethanation potential. Various pretreatment-physical, chemical, and biological-strategies have been developed to overcome the inherent recalcitrance of lignocellulose to anaerobic degradation. Biological pretreatment approach, however, outcompete other pretreatments due to their application in milder conditions, little corrosiveness, and lower byproduct formation. Such pretreatment importantly aids in selectively reducing the lignin content and crystalline nature of the lignocellulosic biomass, which would evidently enhance the hydrolysis and production of monomers for their further anaerobic digestion (AD) for methanation. A variety of applied biological pretreatment strategies comprises microaerobic treatments, ensiling or composting, separation of digestion stages, and pretreatments using various lignocellulolytic fungi alongside. The net energy output through such approaches is substantially more and relatively inexpensive compared to other established chemical and mechanical approaches. The present review highlights the use of biological agents including bacterial, fungal and/or their enzymes which trigger biodegradation of wastes and utilization of lignocellulose for biofuel production. Additionally, the different physical, chemical, and biological pretreatment strategies for biogas yield enhancement are presented.

Fungal disease detection in plants: Traditional assays, novel diagnostic techniques and biosensors.

Fungal diseases in commercially important plants results in a significant reduction in both quality and yield, often leading to the loss of an entire plant. In order to minimize the losses, it is essential to detect and identify the pathogens at an early stage. Early detection and accurate identification of pathogens can control the spread of infection. The present article provides a comprehensive overview of conventional methods, current trends and advances in fungal pathogen detection with an emphasis on biosensors. Traditional techniques are the "gold standard" in fungal detection which relies on symptoms, culture-based, morphological observation and biochemical identifications. In recent times, with the advancement of biotechnology, molecular and immunological approaches have revolutionized fungal disease detection. But the drawback lies in the fact that these methods require specific and expensive equipments. Thus, there is an urgent need for rapid, reliable, sensitive, cost effective and easy to use diagnostic methods for fungal pathogen detection. Biosensors would become a promising and attractive alternative, but they still have to be subjected to some modifications, improvements and proper validation for on-field use.

Drug Targets for Cardiovascular-Safe Anti-Inflammatory: In Silico Rational Drug Studies.

Cyclooxygenase-2 (COX-2) plays an important role in memory consolidation and synaptic activity, the most fundamental functions of the brain. It converts arachidonic acid to prostaglandin endoperoxide H2. In contrast, if over-expressed, it causes inflammation in response to cytokine, pro-inflammatory molecule, and growth factor. Anti-inflammatory agents, by allosteric or competitive inhibition of COX-2, alleviate the symptoms of inflammation. Coxib family drugs, particularly celecoxib, are the most famous anti-inflammatory agents available in the market showing significant inhibitory effect on COX-2 activity. Due to high cardiovascular risk of this drug group, recent researches are focused on the investigation of new safer drugs for anti-inflammatory diseases. Natural compounds, particularly, phytochemicals are found to be good candidates for drug designing and discovery. In the present study, we performed in silico studies to quantitatively scrutinize the molecular interaction of curcumin and its structural analogs with COX-2, COX-1, FXa and integrin αIIbßIII to investigate their therapeutic potential as a cardiovascular-safe anti-inflammatory medicine (CVSAIM). The results of both ADMET and docking study indicated that out of all the 39 compounds studied, caffeic acid had remarkable interaction with proteins involved in inflammatory response. It was also found to inhibit the proteins that are involved in thrombosis, thereby, having the potential to be developed as therapeutic agent.

Dielectrophoretic separation of micron and submicron particles: a review.

This paper provides an overview on separation of micron and submicron sized biological (cells, yeast, virus, bacteria, etc.) and nonbiological particles (latex, polystyrene, CNTs, metals, etc.) by dielectrophoresis (DEP), which finds wide applications in the field of medical and environmental science. Mathematical models to predict the electric field, flow profile, and concentration profiles of the particles under the influence of DEP force have also been covered in this review. In addition, advancements made primarily in the last decade, in the area of electrode design (shape and arrangement), new materials for electrode (carbon, silicon, polymers), and geometry of the microdevice, for efficient DEP separation of particles have been highlighted.

Molecular techniques revealed highly diverse microbial communities in natural marine biofilms on polystyrene dishes for invertebrate larval settlement.

Biofilm microbial communities play an important role in the larval settlement response of marine invertebrates. However, the underlying mechanism has yet to be resolved, mainly because of the uncertainties in characterizing members in the communities using traditional 16S rRNA gene-based molecular methods and in identifying the chemical signals involved. In this study, pyrosequencing was used to characterize the bacterial communities in intertidal and subtidal marine biofilms developed during two seasons. We revealed highly diverse biofilm bacterial communities that varied with season and tidal level. Over 3,000 operational taxonomic units with estimates of up to 8,000 species were recovered in a biofilm sample, which is by far the highest number recorded in subtropical marine biofilms. Nineteen phyla were found, of which Cyanobacteria and Proteobacteria were the most dominant one in the intertidal and subtidal biofilms, respectively. Apart from these, Actinobacteria, Bacteroidetes, and Planctomycetes were the major groups recovered in both intertidal and subtidal biofilms, although their relative abundance varied among samples. Full-length 16S rRNA gene clone libraries were constructed for the four biofilm samples and showed similar bacterial compositions at the phylum level to those revealed by pyrosequencing. Laboratory assays confirmed that cyrids of the barnacle Balanus amphitrite preferred to settle on the intertidal rather than subtidal biofilms. This preference was independent of the biofilm bacterial density or biomass but was probably related to the biofilm community structure, particularly, the Proteobacterial and Cyanobacterial groups.

Proteomic and metabolomic profiles of marine Vibrio sp. 010 in response to an antifoulant challenge.

Vibrio spp. have the ability to form biofilms, which may contribute to the subsequent successful colonization by microfouling and macrofouling organisms. The effects of an antifouling compound, poly-ether B, on Vibrio sp. 010 were investigated using flow cytometry, proteomics, and metabolomics. A 2-D gel-based proteomic analysis was used to identify proteins responsive to poly-ether B treatment. The profiles of biofilm metabolites were analyzed by ultra-performance liquid chromatography-mass spectrometry. Poly-ether B caused a significant reduction in viability. The proteins affected by the treatment were related to nucleotide metabolism, the glyoxylate cycle, and stress responses. Metabolites such as tripeptides, fatty acids, and quorum-sensing molecules were regulated differentially. Down-regulation of proteins and metabolites potentially led to a loss in colonisation ability, thereby affecting the structure of the biofilm. These results suggest that the proteins and metabolites identified may serve as target molecules for potent antifouling compounds.

Effects of poly-ether B on proteome and phosphoproteome expression in biofouling Balanus amphitrite cyprids.

Biofouling is ubiquitous in marine environments, and the barnacle Balanus amphitrite is one of the most recalcitrant and aggressive biofoulers in tropical waters. Several natural antifoulants that were claimed to be non-toxic have been isolated in recent years, although the mechanism by which they inhibit fouling is yet to be investigated. Poly-ether B has shown promise in the non-toxic inhibition of larval barnacle attachment. Hence, in this study, multiplex two-dimensional electrophoresis (2-DE) was applied in conjunction with mass spectrometry to investigate the effects of poly-ether B on barnacle larvae at the molecular level. The cyprid proteome response to poly-ether B treatment was analyzed at the total proteome and phosphoproteome levels, with 65 protein and 19 phosphoprotein spots found to be up- or down-regulated. The proteins were found to be related to energy-metabolism, oxidative stress, and molecular chaperones, thus indicating that poly-ether B may interfere with the redox-regulatory mechanisms governing the settlement of barnacle larvae. The results of this study demonstrate the usefulness of the proteomic technique in revealing the working mechanisms of antifouling compounds.

Poly-ethers from Winogradskyella poriferorum: Antifouling potential, time-course study of production and natural abundance.

A sponge-associated bacterium, Winogradskyella poriferorum strain UST030701-295T was cultured up to 100l for extraction of antifouling bioactive compounds. Five poly-ethers were isolated and partially characterized based on nuclear magnetic resonance (NMR) spectroscopy and mass spectrometry (MS); two of them showed inhibitory effects on biofilm formation of marine bacteria and larval settlement of macro-foulers but did not produce any adverse effects on the phenotypes of zebra fish embryos at a concentration of 5µg ml(-1). The effect of culture duration on the production of the poly-ethers and the bioactivity of the relevant extracts was monitored over a period of 12 days. The total crude poly-ether production increased from day 2 to day 5 and the highest bioactivity was observed on day 3. The poly-ethers were found to be localized in the cellular fraction of the extracts, implying their natural occurrence. The potent bioactivity of these poly-ethers together with their high natural abundance in bacteria makes them promising candidates as ingredients in antifouling applications.

Hydrothermally generated aromatic compounds are consumed by bacteria colonizing in Atlantis II Deep of the Red Sea.

Hydrothermal ecosystems have a wide distribution on Earth and many can be found in the basin of the Red Sea. Production of aromatic compounds occurs in a temperature window of ∼60-150 °C by utilizing organic debris. In the past 50 years, the temperature of the Atlantis II Deep brine pool in the Red Sea has increased from 56 to 68 °C, whereas the temperature at the nearby Discovery Deep brine pool has remained relatively stable at about 44 °C. In this report, we confirmed the presence of aromatic compounds in the Atlantis II brine pool as expected. The presence of the aromatic compounds might have disturbed the microbes in the Atlantis II. To show shifted microbial communities and their metabolisms, we sequenced the metagenomes of the microbes from both brine pools. Classification based on metareads and the 16S rRNA gene sequences from clones showed a strong divergence of dominant bacterial species between the pools. Bacteria capable of aromatic degradation were present in the Atlantis II brine pool. A comparison of the metabolic pathways showed that several aromatic degradation pathways were significantly enriched in the Atlantis II brine pool, suggesting the presence of aromatic compounds. Pathways utilizing metabolites derived from aromatic degradation were also significantly affected. In the Discovery brine pool, the most abundant genes from the microbes were related to sugar metabolism pathways and DNA synthesis and repair, suggesting a different strategy for the utilization of carbon and energy sources between the Discovery brine pool and the Atlantis II brine pool.

Antibacterial and antilarval-settlement potential and metabolite profiles of novel sponge-associated marine bacteria.

In this study, we screened seven novel sponge-associated marine bacteria for their antibacterial and antilarval-settlement activity in order to find possible new sources of non-toxic or less toxic bioactive antifoulants. The anti-bacterial-growth activity of crude extracts of each bacterium was evaluated by the disk-diffusion assay. Extracts of four potent bacteria with high and broad spectra of antibacterial activity were further separated with solvents of different polarities (hexane and ethyl acetate). To evaluate their indirect inhibitive effect on larval settlement, we tested for their antibiofilm formation activity against two of the test bacteria (Vibrio halioticoli and Loktanella hongkongensis) inductive to Hydroides elegans larval settlement. About 60 and 87% of the extracts inhibited biofilm formation by V. halioticoli and by L. hongkongensis respectively. The extracts were also tested for their direct antilarval-settlement activity against the barnacle Balanus amphitrite and the polychaete H. elegans; 87% of the extracts had a strong inhibitive effect on larval settlement of both species. Extracts of two of the isolates completely inhibited larval settlement of B. amphitrite at 70 microg ml(-1) and H. elegans at 60 microg ml(-1). The organic extracts of Winogradskyella poriferorum effectively inhibited the larval settlement of both H. elegans and B. amphitrite and the biofilm formation of the two bacterial species. The metabolites present in the active crude extracts were profiled using GC MS, and the most prevalent metabolites present in all extracts were identified. This study successfully identified potential new sources of antifouling compounds.