Nexus between light and culture media on morphogenesis in Bacopa monnieri and saponin yield thereof.

Bacopa monnieri, a well-documented nootropic plant of high commercial global demand had been explored for its effect in alleviating other diseases and symptoms. This is primarily attributed to different phytocompounds present in the plant. One of the major constituents among them are saponins. However, variation in agro-climatic conditions and choice of germplasm often affect the growth rate and yield of phytocompounds that significantly impact the efficacy of the plant and its extract. Tissue culture has been attempted to improve the yield of phytocompounds but is often restricted by higher cost and scalability. Current study explores the role of commercial hydroponic media 'Leafy 200' vis-à-vis Murashige and Skoog (MS) media, under different color and intensity of lights, on plant morphogenesis, biomass and saponin yield. Blue light induced more shoot differentiation than normal white light. Statistical studies performed using fractional factorial design showed no significant variations in the yield of saponins among the extracts. The study suggests that hydroponic culture to be a sustainable solution and possible substitute to tissue culture that may be exploited for scalable cultivation of the plant.

Comparative evaluation of four triterpenoid glycoside saponins of bacoside A in alleviating sub-cellular oxidative stress of N2a neuroblastoma cells.

OBJECTIVES: To examine the neuroprotective property of triterpenoid glycoside saponins of Bacopa monnieri (L.) Wettst. bacoside A and its components against H2 O2 -induced oxidative stress on neuronal (N2a) cells. METHODS: The cytoprotective effects of individual bacoside A components were evaluated towards oxidative stressed neuronal cells. Bacoside A was screened for neuronal cell viability (MTT assay) and change in intracellular reactive oxygen species (ROS), anti-apoptotic properties and mitochondrial membrane potential (MMP) using fluorescence microscopy. KEY FINDINGS: Different bacoside A components showed decrease in N2a cell viability below 100 (%) after bacoside A concentration of 0.4 mg/ml. Further, cytoprotective effect of optimized dose of bacoside A was analysed for alleviating oxidative stressed, apoptosis and MMP in H2 O2 stressed neuronal cells. Results showed increase in MMP, and decrease in apoptotic induction, without much change in nuclear integrity in stressed neuronal cells. Results showed bacoside A3 and bacopaside II have comparatively higher cytoprotective ability whilst isomer of bacopasponin C, bacopasaponin C and mixture showed comparatively less response. CONCLUSIONS: Amongst four different bacoside A components, bacoside A3 and bacopaside II showed comparatively higher neuroprotective response analysed as higher cell viability and decreased intracellular ROS, suggesting better regulation of cyto-(neuronal) protection of N2a cells.

Zinc oxide nanoparticles (ZnO NP) mediated regulation of bacosides biosynthesis and transcriptional correlation of HMG-CoA reductase gene in suspension culture of Bacopa monnieri.

Bacopa monnieri (L.) Wettst. is a well documented nootropic plant, extensive known for alleviating symptoms of neurological disorder, along with other symptomatic relief. This property is attributed to the active phytocompounds, saponins (bacoside A) present in the plant. However, lack of stringent validation guidelines in most of the countries bring to the market, formulations differing in phytocompounds yield, thereby suggesting possible variation in therapeutic efficacy. The in-vitro suspension cultures of the Bacopa monnieri, provide an ease of scale-up, but regulating saponin yield is a stringent task. The aim of the study is to explore the effects of different concentrations (0, 0.25, 0.50, 0.75 and 1.0 ppm) of zinc oxide nanoparticles (ZnO NP) (24 nm in size), in regulating growth rate, bacoside yield and transcriptional profile of HMG CoA reductasegene in the suspension cells of Bacopa monnieri. Results showed a linear correlation between Bacoside A yield and ZnO NP concentrations with around 2 fold increase in total bacoside A concentration at 1 ppm. Also, ZnO NP supplemented suspension cells showed variation in the specific growth rate. Neuroprotective properties, analyzed using methanolic extracts of suspension cells again obtrude the extract of ZnO NP supplemented (0.75 ppm and 1 ppm) culture for better response in alleviating oxidative stress mediated damage to neuronal cells. ZnO NP supplemented system showed lower expression of HMG CoA reductasegene (the rate limiting step in bacoside A biosynthesis) but higher concentration of bacoside A, suggesting possible role of ZnO NP in isoprenoid pathway than MVA pathways.

Evaluation of physicochemical and biological properties of chitosan/poly (vinyl alcohol) polymer blend membranes and their correlation for Vero cell growth.

The blend membranes with varying weight ratios of chitosan/poly (vinyl alcohol) (CS/PVA) (1:0, 1:1, 1:2.5, 1.5:1, 1.5: 2.5) were prepared using solvent casting method and were evaluated for their potential application in single-use membrane bioreactors (MBRs). The physicochemical properties of the prepared membranes were investigated for chemical interactions (FTIR), surface morphology (SEM), water uptake, protein sorption (qe), ammonia sorption and growth kinetics of Vero cells. CS/PVA blend membrane having weight ratio of 1.5:1 had shown enhanced membrane flexibility, reduced water uptake, less protein sorption and no ammonium sorption compared to CS membrane. This blend membrane also showed comparatively enhanced higher specific growth rate (0.82/day) of Vero cells. Improved physicochemical properties and growth kinetics obtrude CS/PVA (1.5:1) as a potential surface for adhesion and proliferation with possible application in single use membrane bioreactors. Additionally, new insight explaining correlation between water holding (%) of CS/PVA (1.5:1) blend membrane and doubling time (td) of Vero cells is proposed.

Effect of media components on cell growth and bacterial cellulose production from Acetobacter aceti MTCC 2623.

Acetobacter aceti MTCC 2623 was studied as an alternative microbial source for bacterial cellulose (BC) production. Effect of media components on cell growth rate, BC production and cellulose characteristics were studied. FTIR results showed significant variations in cellulose characteristics produced by A. aceti in different media. Results have shown the role of fermentation time on crystallinity ratio of BC in different media. Further, effect of six different media components on cell growth and BC production was studied using fractional factorial design. Citric acid was found to be the most significant media component for cell growth rate (95% confidence level, R(2)=0.95). However, direct role of these parameters on cellulose production was not established (p-value>0.05).

Comparative Molecular docking analysis of DNA Gyrase subunit A in Pseudomonas aeruginosaPAO1.

Pseudomonas aeruginosa is an opportunistic bacterium known for causing chronic infections in cystic fibrosis and chronic obstructive pulmonary disease (COPD) patients. Recently, several drug targets in Pseudomonas aeruginosa PAO1 have been reported using network biology approaches on the basis of essentiality and topology and further ranked on network measures viz. degree and centrality. Till date no drug/ligand molecule has been reported against this targets.In our work we have identified the ligand /drug molecules, through Orthologous gene mapping against Bacillus subtilis subsp. subtilis str. 168 and performed modelling and docking analysis. From the predicted drug targets in PA PAO1, we selected those drug targets which show statistically significant orthology with a model organism and whose orthologs are present in all the selected drug targets of PA PAO1.Modeling of their structure has been done using I-Tasser web server. Orthologous gene mapping has been performed using Cluster of Orthologs (COGs) and based on orthology; drugs available for Bacillus sp. have been docked with PA PAO1 protein drug targets using MoleGro virtual docker version 4.0.2.Orthologous gene for PA3168 gyrA is BS gyrAfound in Bacillus subtilis subsp. subtilis str. 168. The drugs cited for Bacillus sp. have been docked with PA genes and energy analyses have been made. Based on Orthologous gene mapping andin-silico studies, Nalidixic acid is reported as an effective drug against PA3168 gyrA for the treatment of CF and COPD.