OPTIMIZATION OF POMEGRANATE PEEL EXTRACTS FOR THE BIOCONVERSION OF THE ELLAGITANNINS TO ELLAGIC ACID USING ASPERGILLUS NIGER, RHIZOPUS ORYZAE AND MIXED CULTURE

Objective: The present study is aimed at optimization of concentration of substrate (pomegranate peel extract) for the production of Ellagic acid using Aspergillus niger and Rhizopus oryzae. Methods: Test organisms were isolated and identified using standard microbiological techniques. Collected pomegranate peels were dried, grained and used as the substrate for solid-state fermentation. Prepared spore suspension of the test organisms was inoculated and incubated at room temperature. At regular intervals of 24 h, samples were drawn and subjected to analysis of reducing sugar, soluble proteins, hydrolysable tannins and production of extracellular tannase using standard methods. Results: The results of the present investigation demonstrated the optimum substrate concentration for the active conversion of ellagitannins to ellagic was 15g of A. niger, 20g for R. oryzae and 15g for mixed culture. Conclusion: From the current work, it was concluded that solid-state bioprocessing of fruit substrates and fruit wastes using fungi has shown to enrich phenolic antioxidants and improve phytochemical consistency

STANDARDIZATION AND APPLICATION OF PCR TARGETING CHLORELLA SPECIES ISOLATED FROM ENVIRONMENTAL SAMPLES

Objective: Identification of Chlorella species from the environment through 18s ribosomal RNA sequencing. This study was aimed to design primer targeting Chlorella and other closely related algal species targeting 18s ribosomal RNA, ITS1 region. Methods: Sanger sequencing was carried out for the identification of algae up to the genus and species level using an in-house designed primer and optimized PCR conditions. Results: Out of 2 algae samples identified phenotypically, one isolate identified as Chlorella vulgaris and other one identified as Chlorella sorokiniana based on the results of Basic Alignment Search Tool (BLAST). Conclusion: To conclude, this study provided primers with PCR conditions to characterize algal samples through molecular identification with 100% accuracy than the phenotypic method

Lactobacillus rhamnosus blocks inflammatory signaling in vivo via reactive oxygen species generation

Uncontrolled inflammatory responses in the immature gut may play a role in the pathogenesis of many intestinal inflammatory syndromes that present in newborns or children, such as necrotizing enterocolitis (NEC), idiopathic inflammatory bowel diseases (IBD), or infectious enteritis. Consistent with previous reports that murine intestinal function matures over the first 3 weeks of life, we show that inflammatory signaling in the neonatal mouse gut increases during postnatal maturation, with peak responses occurring at 2-3 weeks. Probiotic bacteria can block inflammatory responses in cultured epithelia by inducing the generation of reactive oxygen species (ROS), which inhibit NF-κB activation through oxidative inactivation of the key regulatory enzyme Ubc12. We now report for the first time that the probiotic Lactobacillus rhamnosus GG (LGG) can induce ROS generation in intestinal epithelia in vitro and in vivo. Intestines from immature mice gavage fed LGG exhibited increased GSH oxidation and cullin-1 deneddylation, reflecting local ROS generation and its resultant Ubc12 inactivation, respectively. Furthermore, prefeeding LGG prevented TNF-α-induced intestinal NF-κB activation. These studies indicate that LGG can reduce inflammatory signaling in immature intestines by inducing local ROS generation and may be a mechanism by which probiotic bacteria can prevent NEC in premature infants or reduce the severity of IBD in children

Annexin A1, formyl peptide receptor, and NOX1 orchestrate epithelial repair

N-formyl peptide receptors (FPRs) are critical regulators of host defense in phagocytes and are also expressed in epithelia. FPR signaling and function have been extensively studied in phagocytes, yet their functional biology in epithelia is poorly understood. We describe a novel intestinal epithelial FPR signaling pathway that is activated by an endogenous FPR ligand, annexin A1 (ANXA1), and its cleavage product Ac2-26, which mediate activation of ROS by an epithelial NADPH oxidase, NOX1. We show that epithelial cell migration was regulated by this signaling cascade through oxidative inactivation of the regulatory phosphatases PTEN and PTP-PEST, with consequent activation of focal adhesion kinase (FAK) and paxillin. In vivo studies using intestinal epithelial specific Nox1(–/–IEC) and AnxA1(–/–) mice demonstrated defects in intestinal mucosal wound repair, while systemic administration of ANXA1 promoted wound recovery in a NOX1-dependent fashion. Additionally, increased ANXA1 expression was observed in the intestinal epithelium and infiltrating leukocytes in the mucosa of ulcerative colitis patients compared with normal intestinal mucosa. Our findings delineate a novel epithelial FPR1/NOX1-dependent redox signaling pathway that promotes mucosal wound repair