All-trans retinoic acid ( atRA) is a potent active metabolite of vitamin A that functions as a ligand for the retinoic acid receptor (RAR). Vitamin A, provided from the diet in the form of carotenoids or retinyl esters, is also central to gut immune homeostasis where it coordinates both innate and adaptive immune responses. Secreted by B cells into the mucus layer and lumen of the intestine, sIgA binds to many commensal and pathogenic bacteria and their toxins to prevent their translocation from the gut lumen. These include the release of secretory Immunoglobulin A (sIgA), antimicrobial peptides (AMPs), and downregulation of key nutrients desired by pathogens, such as iron. In order to prevent gut dysbiosis and maintain homeostasis between gut microbes and the host, the host employs several immune-mediated mechanisms. Gut dysbiosis is characterized as expanded opportunistic microbial densities at the expense of reduced commensal microbes. A variety of human diseases are associated with an imbalance in gut microbial composition and function often termed “gut dysbiosis”. These data suggest prophylactic supplementation with a spore-forming Bacillus-containing probiotic may protect against antibiotic-induced dysregulation of host immune responses.Ĭomprised of trillions of microbes, the gut microbiota has co-evolved along with the gut to create a mutualistic relationship with the host centered on gut immune responses and tolerance. Regional responses in mRNA expression of enzymes involved with vitamin A metabolism occurred between antibiotic groups, and intestinal inflammatory markers were mitigated with the probiotic blend. The probiotic blend protected against antibiotic-induced overgrowth of gram-negative bacteria and gammaproteobacteria in the cecum which correlated with host immune responses. Fecal and cecal contents were analyzed for gut microbes, and intestinal tissue was tested for the expression of key enzymes involved in vitamin A metabolism, serum amyloid A, and inflammatory markers in the intestine. Mice were exposed to antibiotics and supplemented with or without the probiotic blend and compared to control mice. Here we aimed to test the efficacy of a non-pathogenic spore-forming Bacillus-species containing a probiotic blend provided during antibiotic therapy on host immune defenses in mice. Probiotics have been tested as a means to counteract the negative effects of antibiotic therapy, but many probiotics are also likely destroyed by antibiotics when taken together. Stable at room temperature for 72 hours once plates are prepared for qPCRġDynamic range is a property of both the assay and template concentration in the sample, as well as in the master mix, thus individual results may vary.Antibiotic therapy is necessary for the treatment of bacterial infections however, it can also disrupt the balance and function of commensal gut microbes and negatively affect the host.Formulated with UNG/dUTP to prevent contamination of carryover PCR products.Compatible with standard or fast cycling for results in less than 50 minutes.Highly reproducible threshold cycles over a broad dynamic range.Exceptional specificity with dual hot-start mechanism.Research labs, biotechnology firms and contract research organizations performing gene expression, DNA quantitation and Chromatin Immunoprecipitation When used with Dual-Lock Taq DNA polymerase, the master mix has been shown to generate consistent performance in preassembled reactions for up to 72 hours. 1 It can be used in either standard or fast cycling mode and is compatible with all qPCR instruments. PowerUp SYBR Green Master Mix provides robust results ranging from 100 ng to 0.1 pg of complementary DNA (cDNA) per reaction.
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