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      Lactobacillus plantarum TWK10 Supplementation Improves Exercise Performance and Increases Muscle Mass in Mice

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          Abstract

          Lactobacillus plantarum ( L. plantarum) is a well-known probiotic among the ingested-microorganism probiotics ( i.e., ingested microorganisms associated with beneficial effects for the host). However, few studies have examined the effects of L. plantarum TWK10 (LP10) supplementation on exercise performance, physical fatigue, and gut microbial profile. Male Institute of Cancer Research (ICR) strain mice were divided into three groups ( n = 8 per group) for oral administration of LP10 for six weeks at 0, 2.05 × 10 8, or 1.03 × 10 9 colony-forming units/kg/day, designated the vehicle, LP10-1X and LP10-5X groups, respectively. LP10 significantly decreased final body weight and increased relative muscle weight (%). LP10 supplementation dose-dependently increased grip strength ( p < 0.0001) and endurance swimming time ( p < 0.001) and decreased levels of serum lactate ( p < 0.0001), ammonia ( p < 0.0001), creatine kinase ( p = 0.0118), and glucose ( p = 0.0151) after acute exercise challenge. The number of type I fibers (slow muscle) in gastrocnemius muscle significantly increased with LP10 treatment. In addition, serum levels of albumin, blood urea nitrogen, creatinine, and triacylglycerol significantly decreased with LP10 treatment. Long-term supplementation with LP10 may increase muscle mass, enhance energy harvesting, and have health-promotion, performance-improvement, and anti-fatigue effects.

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          Extracellular MUC3 mucin secretion follows adherence of Lactobacillus strains to intestinal epithelial cells in vitro.

          Mucins are large complex glycoproteins that protect intestinal mucosal surfaces by limiting access of environmental matter to their epithelial cells. Several mucin genes have been described, including MUC3 that is a membrane associated mucin of the small intestine. Increased MUC3 mRNA transcription is induced by incubation of intestinal epithelial cells with a Lactobacillus strain known to be adherent to them. To determine whether increased epithelial cell MUC3 mucin expression in response to Lactobacillus strains results in increased extracellular secretion of MUC3 mucins and the importance of epithelial cell adherence in modulation of MUC3 mucin expression. HT29 cells grown to enhance expression of MUC3 mucins were incubated with selected Lactobacillus strains. Spent cell culture medium was collected for detection of secreted MUC3 mucins using dot blot immunoassay with a generated MUC3 antibody. Post-incubation HT29 cell RNA was collected for analysis of MUC3 expression by northern blot analysis using a MUC3 cDNA probe. In vitro binding studies using Lactobacillus strains incubated alone or coincubated with enteropathogenic Escherichia coli strain E2348/69 were used for adherence and inhibition of adherence studies, respectively. Lactobacillus strains with minimal ability to adhere to HT29 cells failed to induce upregulation of mucin gene expression. There was a direct correlation between upregulation of MUC3 mucin mRNA expression and extracellular secretion of MUC3 mucin. The same Lactobacillus strains that increased extracellular secretion of MUC3 mucin led to reduced adherence of enteropathogen E coli E2348/69 during coincubation experiments. Probiotic microbes induce MUC3 mucin transcription and translation with extracellular secretion of the MUC3 mucins. Epithelial cell adherence enhances the effects of probiotics on eukaryotic mucin expression.
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            Differential NF-kappaB pathways induction by Lactobacillus plantarum in the duodenum of healthy humans correlating with immune tolerance.

            How do we acquire immune tolerance against food microorganisms and commensal bacteria that constitute the intestinal microbiota? We investigated this by stimulating the immune system of adults with commensal Lactobacillus plantarum bacteria. We studied the in vivo human responses to L. plantarum in a randomized double-blind placebo-controlled cross-over study. Healthy adults ingested preparations of living and heat-killed L. plantarum bacteria. Biopsies were taken from the intestinal duodenal mucosa and altered expression profiles were analyzed using whole-genome microarrays and by biological pathway reconstructions. Expression profiles of human mucosa displayed striking differences in modulation of NF-kappaB-dependent pathways, notably after consumption of living L. plantarum bacteria in different growth phases. Our in vivo study identified mucosal gene expression patterns and cellular pathways that correlated with the establishment of immune tolerance in healthy adults.
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              Antioxidant properties of potentially probiotic bacteria: in vitro and in vivo activities.

              Thirty-four strains of lactic acid bacteria (seven Bifidobacterium, 11 Lactobacillus, six Lactococcus, and 10 Streptococcus thermophilus) were assayed in vitro for antioxidant activity against ascorbic and linolenic acid oxidation (TAA(AA) and TAA(LA)), trolox-equivalent antioxidant capacity (TEAC), intracellular glutathione (TGSH), and superoxide dismutase (SOD). Wide dispersion of each of TAA(AA), TAA(LA), TEAC, TGSH, and SOD occurred within bacterial groups, indicating that antioxidative properties are strain specific. The strains Bifidobacterium animalis subsp. lactis DSMZ 23032, Lactobacillus acidophilus DSMZ 23033, and Lactobacillus brevis DSMZ 23034 exhibited among the highest TAA(AA), TAA(LA), TEAC, and TGSH values within the lactobacilli and bifidobacteria. These strains were used to prepare a potentially antioxidative probiotic formulation, which was administered to rats at the dose of 10(7), 10(8), and 10(9) cfu/day for 18 days. The probiotic strains colonized the colon of the rats during the trial and promoted intestinal saccharolytic metabolism. The analysis of plasma antioxidant activity, reactive oxygen molecules level, and glutathione concentration, revealed that, when administered at doses of at least 10(8) cfu/day, the antioxidant mixture effectively reduced doxorubicin-induced oxidative stress. Probiotic strains which are capable to limit excessive amounts of reactive radicals in vivo may contribute to prevent and control several diseases associated with oxidative stress.
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                Author and article information

                Journal
                Nutrients
                Nutrients
                nutrients
                Nutrients
                MDPI
                2072-6643
                07 April 2016
                April 2016
                : 8
                : 4
                : 205
                Affiliations
                [1 ]Graduate Institute of Sports Science, National Taiwan Sport University, Taoyuan 33301, Taiwan; 1021302@ 123456ntsu.edu.tw (Y.-M.C.); 1021301@ 123456ntsu.edu.tw (Y.-S.C.); 1041302@ 123456ntsu.edu.tw (Y.-J.H.)
                [2 ]Department of Neurosurgery, Taipei Medical University-WanFang Hospital, Taipei City 11696, Taiwan; nsweili@ 123456gmail.com
                [3 ]Department of Orthopedic Surgery, Taipei Medical University-Shuang Ho Hospital, New Taipei City 23561, Taiwan
                [4 ]School of Nutrition and Health Sciences, Taipei Medical University, Taipei 11031, Taiwan
                [5 ]Department of Food Science, Fu Jen Catholic University, Taipei 24205, Taiwan
                [6 ]Department of Food and Nutrition, Providence University, Taichung 43301, Taiwan
                Author notes
                [* ]Correspondence: tytsai@ 123456mail.fju.edu.tw (T.-Y.T.); mfwang@ 123456pu.edu.tw (M.-F.W.); d301090007@ 123456gmail.com or john5523@ 123456ntsu.edu.tw (C.-C.H.); Tel.: +886-2-2905-2539 (T.-Y.T.); +886-4-2632-8001 (ext. 15041) (M.-F.W.); +886-3-328-3201 (ext. 2409) (C.-C.H.)
                [†]

                These authors contributed equally to this work.

                Article
                nutrients-08-00205
                10.3390/nu8040205
                4848674
                27070637
                68a48710-8f56-4b1d-86b8-2bbbbfb84a6c
                © 2016 by the authors; licensee MDPI, Basel, Switzerland.

                This article is an open access article distributed under the terms and conditions of the Creative Commons by Attribution (CC-BY) license ( http://creativecommons.org/licenses/by/4.0/).

                History
                : 23 January 2016
                : 01 April 2016
                Categories
                Article

                Nutrition & Dietetics
                lactobacillus plantarum twk10,exercise performance,forelimb grip strength,gastrocnemius muscles

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