Assessment of impact of varied preconditioning factors, used in isolation and combination, for enhancement of cytokine secretions in stem cell conditioned medium

Stem cell conditioned medium exhibits a huge regenerative potential but low concentration of cytokines, highdevelopment cost, and scalability challenges are major deterrents to product advances. This research studiedimpact on vascular endothelial growth factor (VEGF) and hepatocyte growth factor (HGF) secretions afterpreconditioning human dental pulp stem cells (HDPSCs) with diverse factors, viz., Deferoxamine (250 µM),Epidermal Growth Factor (EGF) (2 ng/ml), Angiotensin I (5mM) and Insulin Transferrin Selenium (ITS) (1%),and Niacinamide (5 mM). Additionally, advantage of using “pooled population” of HDPSCs was ascertained.HDPSCs were incubated under standard culture conditions using optimized growth media. On reaching 80%–85% confluency, media was discarded and fresh serum free media with preconditioning factors, initially oneat a time and subsequently in combination, was added. Spent media, collected after 48 hours of incubation,was used for enzyme-linked immunosorbent assay testing of selected cytokines. In comparison to control,preconditioning factors, used in isolation showed varied but significant upregulation in the secretion of VEGFbut down regulation of HGF. When used in combination, VEGF secretion increased almost 10 times of controlwith no significant change in HGF level. Results demonstrated importance of choosing right preconditioningfactors and right interplay of preconditioning factors to enhance secretions. This research paves the path todevelop an effective and commercially viable conditioned medium that can be used as “biological active” forpharmaceuticals and for developing “customized” serum free medium enriched with selected cytokines

High-density culture of Vero cells in novel perfusion bioreactor system

Viruses are the obligatory intracellular parasites infecting microbes, plants, animals, and humans. They aredead outside host cell but can take-over the host’s cell machinery as soon as they are into it. Several studies oninhibitor compounds have been done for animal viruses including those that are affecting humans, but thereis inadequacy in terms of research and literature for plant viruses that are responsible for losses in crop yieldand quality loss all across the globe. This could be focal point to study plant viruses, their transmission andpathogenicity, and to establish widely used, effective, and advanced approaches for their control. The purposeof this review is to discuss various approaches to control plant viruses that have been developed and applied tocombat plant viral infections. We have divided these approaches into two categories conventional (meristemtip culture, cryotherapy, thermotherapy, and chemotherapy) and advanced (nucleic acid-based approacheslike RNA Silencing, cross-protection, transgenic plants, gene pyramiding, and protein-protein interaction).Moreover, we have discussed and compared the principles, methodologies, advantages, and disadvantages ofeach technique. The approaches have been explored to promote their application in best suited way on variousplants to control viral diseases and to improve food crops quality with increase in production.

Increased activity of goat liver plasma membrane alkaline phosphatase upon release by phosphatidylinositol-specific phospholipase C.

Mammalian alkaline phosphatase (ALP) is attached to the plasma membrane by a unique glycosylphosphatidylinositol (GPI) anchor. The influence of such a complex anchoring device on the enzyme function is not fully understood. Here, we report the effect of cleavage of the GPI anchor on the activity of goat liver plasma membrane ALP (GLPM-ALP). Phosphatidylinositol-specific phospholipase C (PI-PLC) purified from Bacillus cereus was used for the cleavage of the GPI anchor (delipidation) and hence for release of ALP from the membrane. Detergents — octyl-β-D-glucopyranoside (OG) and triton X100 (TX100) were also used for solubilization of ALP from the membrane. Resistance to solubilization by TX100 suggested the association of GPI-ALP with lipid rafts. Solubilization of GLPM-ALP with OG had no effect on the enzyme activity; however, delipidation with PI-PLC resulted in enhanced ALP activity. Kinetic analysis showed catalytic activation of PI-PLC-treated GLPM-ALP with an increase in Vmax (35%) without a significant change in Km. Moreover, this change in Vmax was observed to be independent of pH and buffer. The results suggested the implication of GPI anchor in modulating the catalytic property of GLPM-ALP, thus indicating the role of this special anchoring structure in the enzyme regulation.

Decolorisation of synthetic dyes and textile wastewater using Polyporus rubidus.

Effluent from textile industries were treated with enzyme from white rot fungi isolated from outskirts of Mumbai and identified as Polyporus rubidus in our laboratory. Decolorisation of 4 Reactive dyes commonly found in the effluents such as Reactive bue, Reactive orange, Ramazol black and Congo red was examined by treatment with enzyme from Polyporus rubidus. Treatment of effluent was done in a laboratory scale bioreactor constructed with laccase immobilized Na-alginate beads. Greater than 80% of dyes were degraded within 5 days under stationary incubation conditions. The enzyme had a maxmimum activity of 17.1U after 3 days and was found to be secreted extracellularly by Polyporus rubidus. In this study the Polyporus rubidus has been reported for the first time to have laccase activity offering a promising possibility to develop an easy and cost effective method for degradation of dangerous dyes.

Screening of natural phenolic compounds for potential to inhibit bacterial cell division protein FtsZ.

FtsZ plays an important role in bacterial cell division by polymerizing to form the Z ring at the site of cytokinesis. Phytochemicals are known to disrupt bacterial cell division through inhibition of FtsZ assembly. In the present study phytochemicals like eugenol, trans-cinnamic acid, 4-formyl cinnamic acid, naringenin and caffeic acid were were tested for their potential to inhibit cell division. Effect of these antimicrobial compounds on the growth of E. coli was determined and the inhibition of FtsZ assembly in vitro was investigated. The present study revealed trans-cinnamic acid as the most potent inhibitor of FtsZ assembly.