Body weight, fat weight and organs weight of mice fed the experimental diets for 10 weeks.

<p>C57BL/6J mice were fed a normal diet (ND), a high-sucrose diet (HSD) or a HSD containing <i>L. gasseri</i> BNR17 (10⁹ or 10¹⁰ CFU) for 10 weeks. After measurement of body weight and feed intake, the white adipose tissue, liver, spleen and kidney were removed and weighed. Data represent the means ± SD of eight mice per group. Pairwise <i>t</i>-test:</p>*<p><i>P</i><0.05,</p>**<p><i>P</i><0.01,</p>***<p><i>P</i><0.001 <i>versus</i> the ND group;</p>#<p><i>P</i><0.05,</p>##<p><i>P</i><0.01,</p>###<p><i>P</i><0.001 <i>versus</i> the HSD group.</p

<i>L. gasseri</i> BNR17 affects mRNA expression in the liver.

<p>C57BL/6J mice were given ND, HSD, or HSD containing BNR17 (10⁹ or 10¹⁰ CFU) for 10 weeks. The liver was then removed and mRNA expression was measured by real-time RT-PCR using β-actin as a housekeeping gene. Data represent the means ± SD. Pairwise <i>t</i>-test: *<i>P</i><0.05, **P<0.01, <i>versus</i> the ND group; ^#<i>P</i><0.05, ^##<i>P</i><0.01 <i>versus</i> the HSD group.</p

<i>L. gasseri</i> BNR17 affects mRNA expression in white adipose tissue.

<p>C57BL/6J mice were given ND, HSD, or HSD containing BNR17 (10⁹ or 10¹⁰ CFU) for 10 weeks. The white adipose tissue was then removed and mRNA expression was measured by real-time RT-PCR using β-actin as a housekeeping gene. Data represent the means ± SD. Pairwise <i>t</i>-test: *<i>P</i><0.05, **<i>P</i><0.01 <i>versus</i> the ND group; ^#<i>P</i><0.05 <i>versus</i> the HSD group.</p

<i>L. gasseri</i> BNR17 affects endocrine hormones.

<p>C57BL/6J mice were given ND, HSD, or HSD containing BNR17 (10⁹ or 10¹⁰ CFU) for 10 weeks. Serum was obtained by centrifugation of whole blood and analyzed. GIP, glucose-dependent insulinotropic polypeptide; GLP, glucagon-like peptide. Data represent the means ± SD. Wilcoxon rank-sum test: *<i>P</i><0.05, **<i>P</i><0.01, ***<i>P</i><0.001 <i>versus</i> the ND group; ^#<i>P</i><0.05, ^##<i>P</i><0.01 <i>versus</i> the HSD group.</p

Primer sequences of mouse mRNA.

<p>PPARα<b>,</b> peroxisome proliferator-activated receptor α; PPARδ, peroxisome proliferator-activated receptor δ; CPT, carnitine palmitoyl-transferase; ACO, acyl CoA oxidase; UCP3, uncoupling proteins3; GLUT4, glucose transporter 4; SREBP-1c, sterol regulatory element-binding protein-1c; ACC, acetyl-CoA carboxylase; FAS, fatty acid synthetase; PPARγ<b>,</b> peroxisome proliferator-activated receptor γ; LPL, lipoprotein lipase; TNF-α, tumor necrosis factor-α.</p

<i>L. gasseri</i> BNR17 supplementation decreases high-sucrose diet-induced body weight gain and fat mass accumulation.

<p>(A) Change in body weight, (B) change in food intake, (C) representative CT scanning images of abdominal (left) and whole body (right) fat accumulation (in black) at 10 weeks (D) correspond to the volume of subcutaneous and abdominal fat, (E) representative adipose tissue-staining images in mice of four groups, (F) adipocyte mean area (µm²). Data represent the means ± SD. Pairwise <i>t</i>-test: *<i>P</i><0.05, **<i>P</i><0.01, ***<i>P</i><0.001 <i>versus</i> the ND group; ^#<i>P</i><0.05, ^##<i>P</i><0.01, ^###<i>P</i><0.001 <i>versus</i> the HSD group.</p

A <i>Salmonella typhimurium</i> ghost vaccine induces cytokine expression <i>in vitro</i> and immune responses <i>in vivo</i> and protects rats against homologous and heterologous challenges - Fig 5

<p><b>Serum bactericidal activities of rats vaccinated with STG against (a) <i>S</i>. <i>typhpimurium</i> or (b) <i>S</i>. <i>enteritidis</i>.</b> Data were expressed as means ± standard errors of the means. The asterisks indicate significant differences between serum bactericidal activities of the vaccinated and non-vaccinated groups. ** = <i>P</i> < 0.01; * = <i>P</i> < 0.05.</p

Cell viability and pro‐ and anti‐inflammatory cytokine production of STG.

<p>To determine cytotoxicity, murine macrophages (RAW 264.7) were exposed to PBS buffer, LPS purified from <i>S</i>. <i>typhimurium</i>, FKC, wild-type bacterial cells (ST) and NaOH-induced STG, respectively (bars). At 24 h post-exposure, macrophages were collected for analysis of cell viability using Cell counting Kit-8. Absorbance was measured at 450 nm and all experiments were performed in triplicate. Cytotoxic activity is expressed as the percentage of cell viability by the formula described in Materials and Methods. At given time post‐exposure with PBS buffer, LPS purified from <i>S</i>. <i>typhimurium</i>, FKC, ST and STG, respectively (bars), macrophages were collected for analysis of mRNA expression for pro- and anti-inflammatory cytokines and factor (TNF‐α, IL‐1β, iNOS, IL‐6 and IL‐10) using RT-qPCR. Data are representative of triplicate experiments with each sample run in triplicate. All statistical analyses were performed with one-way ANOVA using SPSS software (version 14.0) and data were expressed as mean ± standard error of the mean. *** = <i>P</i> < 0.001; ** = <i>P</i> < 0.01; * = <i>P</i> < 0.05 (significant difference from PBS) and ### = <i>P</i> < 0.001; ## = <i>P</i> < 0.01; # = <i>P</i> < 0.05 (significant difference from LPS).</p

The Natural Anticancer Agent Plumbagin Induces Potent Cytotoxicity in MCF-7 Human Breast Cancer Cells by Inhibiting a PI-5 Kinase for ROS Generation

<div><p>Drug-induced haploinsufficiency (DIH) in yeast has been considered a valuable tool for drug target identification. A plant metabolite, plumbagin, has potent anticancer activity via reactive oxygen species (ROS) generation. However, the detailed molecular targets of plumbagin for ROS generation are not understood. Here, using DIH and heterozygous deletion mutants of the fission yeast <em>Schizosaccharomyces pombe</em>, we identified 1, 4-phopshatidylinositol 5-kinase (PI5K) its3 as a new molecular target of plumbagin for ROS generation. Plumbagin showed potent anti-proliferative activity (GI₅₀; 10 µM) and induced cell elongation and septum formation in wild-type <em>S. pombe</em>. Furthermore, plumbagin dramatically increased the intracellular ROS level, and pretreatment with the ROS scavenger, N-acetyl cysteine (NAC), protected against growth inhibition by plumbagin, suggesting that ROS play a crucial role in the anti-proliferative activity in <em>S. pombe</em>. Interestingly, significant DIH was observed in an its3-deleted heterozygous mutant, in which ROS generation by plumbagin was higher than that in wild-type cells, implying that its3 contributes to ROS generation by plumbagin in this yeast. In MCF7 human breast cancer cells, plumbagin significantly decreased the level of a human ortholog, 1, 4-phopshatidylinositol 5-kinase (PI5K)-1B, of yeast its3, and knockdown of PI5K-1B using siPI5K-1B increased the ROS level and decreased cell viability. Taken together, these results clearly show that PI5K-1B plays a crucial role in ROS generation as a new molecular target of plumbagin. Moreover, drug target screening using DIH in <em>S. pombe</em> deletion mutants is a valuable tool for identifying molecular targets of anticancer agents.</p> </div

Protection of STG against homologous and heterologous challenges.

<p>Bacterial loads in liver, lung, spleen and kidney homogenates after (a-d) a homologous challenge with <i>S</i>. <i>typhimurium</i> or (e-h) a heterologous challenge with <i>S</i>. <i>enteritidis</i>. Results are expressed as means ± standard errors of the means. The asterisks indicate significant differences between the bacterial clearance of the vaccinated and non-vaccinated groups. ** = <i>P</i> < 0.01; * = <i>P</i> < 0.05.</p