Phytochemical screening, antimicrobial, anti-inflammatory and anti-cancerous activities of ethanol and hexane extracts of Urochloa ramosa

Urochloa ramosa is known for its environmental benefits such as stabilization and reclamation of polluted soils, in agriculture to control root-knot nematodes infecting crops, in medicine to treat cardiovascular diseases, duodenal ulcer, hyperglycemia, nephritis and snake bites. Qualitative analyses of phytochemicals in ethanol and hexane extracts were performed by standard methods. In vitro anti-microbial assay was performed against gram positive bacteria viz., Bacillus subtilis and Staphylococcus aureus, Gram negative bacteria viz., Pseudomonas aeruginosa and Escherichia coli and fungus Candida albicans by disc diffusion method. Hexane extract of Urochloa ramosa was found to be 70% effective against Candida albicans indicating potent antifungal property. In vitro anti-inflammatory activity was performed by albumin denaturation assay, proteinase inhibition activity assay and membrane stabilization assay using various concentrations of extracts with Aspirin and Diclofenac sodium as standard. Heat induced protein denaturation was considerably prevented by ethanol and hexane extract at concentrations between 200-500 µg/ml resulting in 60 and 62% inhibition respectively. Heat induced haemolysis of erythrocyte was remarkably inhibited (59 and 68 % respectively) at concentration of 500 µg/ml. 62 and 65 % Hypotonicity induced haemolysis was also inhibited between concentration of 300-500 µg/ml in both extracts respectively. Inhibition of formation of new blood vessels by Chorioallantoic membrane (CAM) assay proved anti-angiogenic effects of extracts. Purification, characterization and structural elucidation of bioactive molecules present in ethanol and hexane extracts are needed to be explored further for assessment of better biological activites than crude extract

Classical Electron Model with Negative Energy Density in Einstein-Cartan Theory of Gravitation

Experimental result regarding the maximum limit of the radius of the electron \sim 10^{-16} cm and a few of the theoretical works suggest that the gravitational mass which is a priori a positive quantity in Newtonian mechanics may become negative in general theory of relativity. It is argued that such a negative gravitational mass and hence negative energy density also can be obtained with a better physical interpretation in the framework of Einstein-Cartan theory.Comment: 12 Latex pages, added refs and conclusion

PROBING THE BROAD-SPECTRUM THERAPEUTIC POTENTIAL OF AIP II MIMICS TO COMBAT LYSOZYME MEDIATED STAPHYLOCOCCAL INVASION ON CONTACT LENS

Objective: Molecular recognition of AIP II mimics as a global inhibitor against the AgrC variants and to undertake a real-time clinical applications to treat the lysozyme mediated (tear protein) S. aureus adherence on contact lens.Methods: Structure activity relationship of the mimic peptides against the receptor AgrC variants were studied to score the global inhibitor. Further, the activity of the mimics as inhibitors was validated through in vitro and in vivo analysis.Results: Inhibition of agr expression of interstrains by the mimic compounds gained insight to recognize a global inhibitorâ€. Further, the in vitro data were designed in such a way to provide a natural eye environment (artificial tears) to see the effect of mAIP IIa (IC50) showed a greater significance of eradicating the clinical isolate, S. aureus biofilm and various other secreted toxins.Conclusion: The mimic peptide (mAIPII a) revealed to be a potential mimic of AIPII to show a broad range inhibition of all AgrC variants without any cytotoxic effects.Â

DESIGN, SYNTHESIS AND VALIDATION OF CqsS RECEPTOR AGONIST TO MODULATE THE QUORUM SENSING CIRCUIT OF VIBRIO CHOLERAE–A MOLECULAR MIMIC THERAPY

Objective: Our earlier studies have characterized a compound from Melia dubia leaves, 4-ethyl resorcinol as an agonist of the Quorum Sensing (QS) receptor, CqsS agonist that had reverse engineered the QS circuit of V. cholerae from low-density to high-density state to effectively inhibit biofilm and enhance the production of protease to detach itself form the host tissue. So, the objective of this study was to synthesize structural derivatives of 4-ethyl resorcinol, to enhance its activity and to down regulate the host cell toxicity was validated.Methods: The antimicrobial (cell-density) and anti-virulent (protease, hemolysis, stress responds and biofilm inhibition) properties of the structural derivatives of 4-ethyl resorcinol were performed.Results: The results indicate that the compound has up surged protease expression along with a remarkable decrease in hemolytic activity at the 7th hour. The stress responds in treated culture has a higher survival rate while the bacterial cells in the control succumb to stress stating the potential of the drug to induce HCD (high cell density) condition in the LCD (low cell density) state. From CLSM analysis we state that there was significant amount of dead colonies and disrupted biofilm in the respective treated cultureConclusion: This could possibly open up a direction to curb the bacterial biofilm formation and may also turn out to be a potent drug against treating Vibrio cholerae infection at an early pointÂ

IL1B Induced Smad 7 Negatively Regulates Gastrin Expression

BACKGROUND: Helicobacter pylori elicited IL1B is one of the various modulators responsible for perturbation of acid secretion in gut. We have earlier reported that IL1B activated NFkB downregulates gastrin, a major modulator of acid secretion. However, we hypothesized that regulation of gastrin by IL1B would depend on the cell's ability to integrate inputs from multiple signaling pathways to generate appropriate biological response. PRINCIPAL FINDING: In this study, we report that IL1B induces Smad 7 expression by about 4.5 fold in gastric carcinoma cell line, AGS. Smad 7 resulted in transcriptional repression of gastrin promoter by about 6.5 fold when co-transfected with Smad 7 expression vector and gastrin-promoter luciferase in AGS cells. IL1B inhibited phosphorylation of Smad 3 and subsequently interfered with nuclear translocation of the positive Smad complex, thus occluding it off the gastrin promoter. IL1B promoter polymorphisms (-511T/-31C IL1B) are known to be associated with H. pylori associated gastro-duodenal ulcer. We observed that IL1B expressed from -31T promoter driven IL1B cDNA elicited 3.5 fold more Smad 7 than that expressed from the IL1B-31C variant in AGS cells. This differential activation of Smad 7 by IL1B promoter variants translated into differential downregulation of gastrin expression. We further analyzed Smad 7, NFkB, IL1B and gastrin expression in antral gut biopsy samples of patients with H. pylori associated duodenal ulcer and normal individuals. We observed that individuals with duodenal ulcer had significantly lower levels of IL1B, Smad 7, NFkB and corresponding higher level of gastrin expression. CONCLUSION: Pro-inflammatory cytokine IL1B repress gastrin expression by activating Smad 7 and subsequent inhibition of nuclear localization of Smad 3/4 complex. Polymorphic promoter variants of IL1B gene can modulate the IL1B expression which resulted in differential activation Smad 7 and consequent repression of gastrin expression, respectively. Analysis of H. pylori infected duodenal ulcer patient's gut biopsy samples also supported this observation

Systems genetics identifies a role for Cacna2d1 regulation in elevated intraocular pressure and glaucoma susceptibility

Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article�s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article�s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/.Glaucoma is a multi-factorial blinding disease in which genetic factors play an important role. Elevated intraocular pressure is a highly heritable risk factor for primary open angle glaucoma and currently the only target for glaucoma therapy. Our study helps to better understand underlying genetic and molecular mechanisms that regulate intraocular pressure, and identifies a new candidate gene, Cacna2d1, that modulates intraocular pressure and a promising therapeutic, pregabalin, which binds to CACNA2D1 protein and lowers intraocular pressure significantly. Because our study utilizes a genetically diverse population of mice with known sequence variants, we are able to determine that the intraocular pressure-lowering effect of pregabalin is dependent on the Cacna2d1 haplotype. Using human genome-wide association study (GWAS) data, evidence for association of a CACNA2D1 single-nucleotide polymorphism and primary open angle glaucoma is found. Importantly, these results demonstrate that our systems genetics approach represents an efficient method to identify genetic variation that can guide the selection of therapeutic targets

Enzymatic Blockade of the Ubiquitin-Proteasome Pathway

Ubiquitin-dependent processes control much of cellular physiology. We show that expression of a highly active, Epstein-Barr virus-derived deubiquitylating enzyme (EBV-DUB) blocks proteasomal degradation of cytosolic and ER-derived proteins by preemptive removal of ubiquitin from proteasome substrates, a treatment less toxic than the use of proteasome inhibitors. Recognition of misfolded proteins in the ER lumen, their dislocation to the cytosol, and degradation are usually tightly coupled but can be uncoupled by the EBV-DUB: a misfolded glycoprotein that originates in the ER accumulates in association with cytosolic chaperones as a deglycosylated intermediate. Our data underscore the necessity of a DUB activity for completion of the dislocation reaction and provide a new means of inhibition of proteasomal proteolysis with reduced cytotoxicity.National Institutes of Health (U.S.)EMBO (long term Fellowship 2008-379)Boehringer Ingelheim Fond

NF-kappaB Mediated Transcriptional Repression of Acid Modifying Hormone Gastrin

Helicobacter pylori is a major pathogen associated with the development of gastroduodenal diseases. It has been reported that H. pylori induced pro-inflammatory cytokine IL1B is one of the various modulators of acid secretion in the gut. Earlier we reported that IL1B-activated NFkB down-regulates gastrin, the major hormonal regulator of acid secretion. In this study, the probable pathway by which IL1B induces NFkB and affects gastrin expression has been elucidated. IL1B-treated AGS cells showed nine-fold activation of MyD88 followed by phosphorylation of TAK1 within 15 min of IL1B treatment. Furthermore, it was observed that activated TAK1 significantly up-regulates the NFkB subunits p50 and p65. Ectopic expression of NFkB p65 in AGS cells resulted in about nine-fold transcriptional repression of gastrin both in the presence and absence of IL1B. The S536A mutant of NFkB p65 is significantly less effective in repressing gastrin. These observations show that a functional NFkB p65 is important for IL1B-mediated repression of gastrin. ChIP assays revealed the presence of HDAC1 and NFkB p65 along with NCoR on the gastrin promoter. Thus, the study provides mechanistic insight into the IL1B-mediated gastrin repression via NFk

Pooled extracellular receptor-ligand interaction screening using CRISPR activation.

Extracellular interactions between cell surface receptors are necessary for signaling and adhesion but identifying them remains technically challenging. We describe a cell-based genome-wide approach employing CRISPR activation to identify receptors for a defined ligand. We show receptors for high-affinity antibodies and low-affinity ligands can be unambiguously identified when used in pools or as individual binding probes. We apply this technique to identify ligands for the adhesion G-protein-coupled receptors and show that the Nogo myelin-associated inhibitory proteins are ligands for ADGRB1. This method will enable extracellular receptor-ligand identification on a genome-wide scale