Project description:Chemotherapy may cause diarrhoea, which may be associated with treatment delay and infections. The purpose of the study is to investigate whether oral supplementation with lactobacilli will alleviate chemotherapy related diarrhoea. Patients diagnosed with advanced colorectal cancer and who will receive chemotherapy will be randomly assigned to receive either lactobacilli or placebo during chemotherapy. The study is a prospective, multicenter, randomized, double-blind, placebo-controlled study. The primary outcome measure is frequency of moderate/severe diarrhoea. The study will also address safety and tolerability of chemotherapy, response to chemotherapy, and serum growth factor levels.
Project description:We performed RNA-seq and Ribo-seq analyses to elucidate the translation in seeds at 85 and 115 DAF. We also completed a data-independent acquisition (DIA)-based proteomic analysis, while also examining relevant lipid metabolites.
Project description:Mass spectrometry analyses for determining whether 13C615N2-labeled lysines contained in the elafin (innate peptide highly expressed in lower female genital tract and down-regulated in case of HPV infection) were found in the amino acid sequence of some proteins of Lactobacilli, supporting the fundamental role of innate secretion in the growth/survival of acid lactic bacteria (L. crispatus, L. jensenii and L. iners).
Project description:Background & Objectives: Identification of news targets for metabolic diseases treatment or prevention was required. In this context, FIAF/ANGPTL4 appeared as a crucial regulator of energy homeostasis. Lactobacilli are often considered to display beneficial effect for their hosts, acting on regulatory pathway. The aim of the present work was to study the effect of several lactobacilli strains on Fiaf gene expression in human intestinal epithelial cells (IECs) and on mice tissues to decipher the underlying mechanisms. Subjects & Methods: Nineteen lactobacilli strains have been tested on HT-29 cells for their ability to regulate Fiaf gene expression by RT-qPCR. In order to determine regulated pathways, we analyzed the whole genome transcript of IECs. We then validate in vivo bacterial effect using C57BL/6 mono-colonized mice fed with normal chow. Results: We identified one strain (L. rhamnosus CNCMI-4317, p<0,001) modulating Fiaf expression in IECs. This regulation relies potentially on bacterial surface-exposed molecules and seems to be PPAR-γ independent and PPAR-α dependent. Functional analysis revealed that most of the significant different expressed genes affected by this strain were involved in cellular function and maintenance, lymphoid tissues structure and development or lipids metabolism. The regulation of immune system and lipids and carbohydrates metabolism was also confirmed by overrepresentation of Gene Ontology terms analysis. In vivo, the strain increased FIAF protein (p<0,05) in the serum and tend to up regulate it in distal small intestine (p=0,14). Moreover, the observed induction of IL-7 (p<0,05) suggested a role on immune cells regulation. Conclusion: We showed that the strain induce Fiaf expression in human IECs and in mice intestine. This bacterial effect is accompanied by modulation of immune response and metabolism providing such a bacterial strain an interesting candidate to modulate metabolic and low grade inflammation disorders.
Project description:Short-chain fructooligosaccharides (scFOS) and other prebiotics are used to selectively stimulate the growth and activity of lactobacilli and bifidobacteria in the colon. However, there is little information on the mechanisms whereby prebiotics exert their specific effects upon such microorganisms. To study the genomic basis of scFOS metabolism in Lactobacillus plantarum WCFS1, two-colour microarrays were used to screen for differentially expressed genes when grown on scFOS as compared to glucose (not a prebiotic). A significant up-regulation (8 to 60-fold) was observed with a set of only five genes located in a single locus and predicted to encode a sucrose phosphoenolpyruvate transport system (PTS), a â-fructofuranosidase, a fructokinase, an á-glucosidase and a sucrose operon repressor. Several other genes were slightly overexpressed, including pyruvate dehydrogenase. For the latter, no detectable activity in L. plantarum under various growth conditions, has been previously reported. A mannose-PTS likely to encode glucose uptake was 50-fold down-regulated as well as, to a lower extent, other PTS. Chemical analysis of the different moieties of scFOS that were depleted in the growth medium revealed that the trisaccharide 1-kestose present in scFOS was preferentially utilized, in comparison with the tetrasaccharide nystose and the pentasaccharide fructofuranosylnystose. The main end-products of scFOS fermentation were lactate and acetate. This is the first example in lactobacilli of the association of a sucrose PTS and a â-fructofuranosidase for scFOS degradation. Keywords: comparative transcriptomics, carbohydrate metabolism
Project description:Lactobacilli are probiotics that, among other health promoting effects, have been ascribed immunostimulating and virus preventive properties. Certain lactobacilli species have been shown to possess strong IL-12 inducing properties. As IL-12 production depends on the up-regulation of type I interferons, we hypothesized that the strong IL-12 inducing capacity of L. acidophilus NCFM in murine bone marrow derived DC is caused by an up-regulation of IFN-β, which subsequently stimulates the induction of IL-12 and the dsRNA binding toll like receptor (TLR)-3. The expression of the genes encoding IFN-β, IL-12, IL-10 and TLR-3 in DC upon stimulation with L. acidophilus NCFM was measured. L. acidophilus NCFM induced a much stronger expression of ifn-β, il-12 and il-10 compared to the synthetic dsRNA ligand Poly I:C, whereas the levels of expressed tlr-3 were similar. By the use of whole genome microarray gene expression, we investigated whether other genes related to the viral defence were up-regulated in DC upon stimulation with L. acidophilus NCFM and found that various virus defence related genes, both early and late, were among the strongest up-regulated genes. The IFN-β stimulating capability was also detected in another L. acidophilus strain, but was not a property of other probiotic bacteria tested (B. bifidum and E. coli nissle).The IFN-β inducing capacity was markedly reduced in TLR-2 -/- DCs, dependent on endocytosis and the major cause of the induction of il-12 and tlr-3 in L. acidophilus NCFM stimulated cells. Collectively, our results reveal that certain lactobacilli trigger the expression of viral defence genes in DC in a TLR-2 manner through induction of IFN- β.
Project description:Lactobacilli are probiotics that, among other health promoting effects, have been ascribed immunostimulating and virus preventive properties. Certain lactobacilli species have been shown to possess strong IL-12 inducing properties. As IL-12 production depends on the up-regulation of type I interferons, we hypothesized that the strong IL-12 inducing capacity of L. acidophilus NCFM in murine bone marrow derived DC is caused by an up-regulation of IFN-β, which subsequently stimulates the induction of IL-12 and the dsRNA binding toll like receptor (TLR)-3. The expression of the genes encoding IFN-β, IL-12, IL-10 and TLR-3 in DC upon stimulation with L. acidophilus NCFM was measured. L. acidophilus NCFM induced a much stronger expression of ifn-β, il-12 and il-10 compared to the synthetic dsRNA ligand Poly I:C, whereas the levels of expressed tlr-3 were similar. By the use of whole genome microarray gene expression, we investigated whether other genes related to the viral defence were up-regulated in DC upon stimulation with L. acidophilus NCFM and found that various virus defence related genes, both early and late, were among the strongest up-regulated genes. The IFN-β stimulating capability was also detected in another L. acidophilus strain, but was not a property of other probiotic bacteria tested (B. bifidum and E. coli nissle).The IFN-β inducing capacity was markedly reduced in TLR-2 -/- DCs, dependent on endocytosis and the major cause of the induction of il-12 and tlr-3 in L. acidophilus NCFM stimulated cells. Collectively, our results reveal that certain lactobacilli trigger the expression of viral defence genes in DC in a TLR-2 manner through induction of IFN- β. Experiment Overall Design: In the experiment Lactobacillus NCFM were added to murine dendritic cells and stimulated for 4, 10 or 18 hours. These were compared to control experiment at the same timepoints. Experiments were run in triplicates except for control 10h and control 18h which were only in duplicate, giving a total of 16 arrays.
Project description:Lactobacillus acidophilus (L. acidophilus) is one of major commensal bacteria in chicken intestine. Lactobacilli have been shown to exert health-promoting and immunostimulating activities. To examine the immunostimulating effects of probiotics, chicken cecal tonsil cells and splenocytes were stimulated in vitro with DNA, peptidoglycan, and cell envelope extracted from L. acidophilus. These bacterial constituents are known to stimulate innate defence mechanisms. Several gene clusters including chemokines and their receptors, antigen processing and presentations, apoptosis related genes were identified in the present study. These differentially expressed genes are candidates for detailed hypothesis-driven investigation of genes elucidating molecular/cellular mechanisms of effects of commensal bacteria on gut immune system in chickens. Keywords: Gene expression profiling of stimulated and unstimulated cells