No abstract

The composition of the colonic microbiota of 91 northern Europeans was characterized by fluorescent in situ hybridization using 18 phylogenetic probes. On average 75% of the bacteria were identified, and large interindividual variations were observed. Clostridium coccoides and Clostridium leptum were the dominant groups (28.0% and 25.2%), followed by the Bacteroides (8.5%). According to principal component analysis, no significant grouping with respect to geographic origin, age, or gender was observed.For the past 10 years, the progress made in molecular technologies has given rise to new ways to explore the human colonic microbiota. Investigations based on 16S rRNA sequences have revealed the presence of hundreds of molecular species, the majority uncultivated and/or not yet cultivated, unique to their host and with few species shared between two individuals (5, 10, 13). Fluorescent in situ hybridization (FISH) (1) of the 16S rRNA has shown that the species diversity comprised less than 20 dominant phylogenetic groups (2-4, 7). However, the molecular analysis has so far been restricted to limited cohorts of individuals, recruited within a single geographic region or country (3,4,7,8,14). In this study, we characterized the fecal microbiota of 91 individuals from five northern European countries to provide a large-scale molecular analysis of the normal colonic microbiota in healthy humans. Multivariate data analysis was performed in order to seek a possible link between the composition of the fecal microbiota and age, gender, or geographic origin parameters.

microRNAs regulate the expression of over 60% of protein coding genes by targeting their mRNAs to AGO2-containing complexes in the cytoplasm and promoting their translational inhibition and/or degradation. There is little evidence so far for microRNA-mediated regulation of other classes of non-coding RNAs. Here we report that microRNA-9 (miR-9) regulates the expression of the Metastasis Associated Lung Adenocarcinoma Transcript 1 (MALAT-1), one of the most abundant and conserved long non-coding RNAs. Intriguingly, we find that miR-9 targets AGO2-mediated regulation of MALAT1 in the nucleus. Our findings reveal a novel direct regulatory link between two important classes of non-coding RNAs, miRs and lncRNAs, and advance our understanding of microRNA functions.

Osteoblast differentiation and bone formation (osteogenesis) are regulated by transcriptional and post-transcriptional mechanisms. Recently, microRNAs (miRNAs) were identified as novel key regulators of human stromal (skeletal, mesenchymal) stem cells (hMSC) differentiation. Here, we identified miRNA-34a (miR-34a) and its target protein networks as modulator of osteoblastic (OB) differentiation of hMSC. miRNA array profiling and further validation by quantitative RT-PCR revealed that miR-34a was upregulated during OB differentiation of hMSC, and in situ hybridization confirmed its OB expression in vivo. Overexpression of miR-34a inhibited early commitment and late OB differentiation of hMSC in vitro, whereas inhibition of miR-34a by anti-miR-34a enhanced these processes. Target prediction analysis and experimental validation confirmed Jagged1 (JAG1), a ligand for Notch 1, as a bona fide target of miR-34a. siRNAmediated reduction of JAG1 expression inhibited OB differentiation. Moreover, a number of known cell cycle regulator and cell proliferation proteins, such as cyclin D1, cyclin-dependent kinase 4 and 6 (CDK4 and CDK6), E2F transcription factor three, and cell division cycle 25 homolog A were among miR-34a targets. Furthermore, in a preclinical model of in vivo bone formation, overexpression of miR-34a in hMSC reduced heterotopic bone formation by 60%, and conversely, in vivo bone formation was increased by 200% in miR-34a-deficient hMSC. miRNA34a exhibited unique dual regulatory effects controlling both hMSC proliferation and OB differentiation. Tissue-specific inhibition of miR-34a might be a potential novel therapeutic strategy for enhancing in vivo bone formation. STEM CELLS 2014;32:902-912

It was previously shown that enhanced nisin resistance in some mutants was associated with increased expression of three genes, pbp2229, hpk1021, and lmo2487, encoding a penicillin-binding protein, a histidine kinase, and a protein of unknown function, respectively. In the present work, we determined the direct role of the three genes in nisin resistance. Interruption of pbp2229 and hpk1021 eliminated the nisin resistance phenotype. Interruption of hpk1021 additionally abolished the increase in pbp2229 expression. The results indicate that this nisin resistance mechanism is caused directly by the increase in pbp2229 expression, which in turn is brought about by the increase in hpk1021 expression. We also found a degree of cross-protection between nisin and class IIa bacteriocins and investigated possible mechanisms. The expression of virulence genes in one nisin-resistant mutant and two class IIa bacteriocin-resistant mutants of the same wild-type strain was analyzed, and each mutant consistently showed either an increase or a decrease in the expression of virulence genes (prfA-regulated as well as prfA-independent genes). Although the changes mostly were moderate, the consistency indicates that a mutant-specific change in virulence may occur concomitantly with bacteriocin resistance development.Nisin and the class IIa bacteriocins (also called pediocin-like bacteriocins) are antimicrobial peptides that are produced by lactic acid bacteria and that have the greatest potential as biopreservatives for food. One of the main target organisms in this context is Listeria monocytogenes, a food-borne pathogen that causes severe human illness as well as economic losses for the food industry.Nisin exerts its antimicrobial action by forming pores in the cytoplasmic membrane through an interaction with the peptidoglycan precursor lipid II (for a recent review, see reference 17). Enhanced nisin resistance in L. monocytogenes generally constitutes less than a 10-fold increase in the MIC. Nisin resistance in several, but not all, spontaneous mutants of L.

No abstract

SummaryThe clinical use of dendritic cells (DCs) to induce antigen-specific immune tolerance has been hampered by the lack of a widely acknowledged method for generating human regulatory DCs but even more so by the non-existence of reliable markers. Thus, we set out to find reliable markers that can be measured with simple methods to identify regulatory DCs that are applicable for future clinical studies. Human DCs were generated from peripheral blood monocytes in the presence of 1a,25-dihydroxyvitamin D3 (VD3), which gave rise to a phenotype that resembles immature DCs, with the exception of high CD14 and reduced CD1a on the cell surface. These VD3-treated DCs exert a long-lasting inefficient T cell stimulation and induce T cell hyporesponsiveness with regulatory potential. Importantly, such VD3-treated DCs were readily distinguishable from untreated DCs by low levels of interleukin-23 secretion and low expression of miR-155 upon exposure to maturation stimuli. Furthermore, VD3-treated DCs showed over-expression of miR-378. All these features can be used as robust markers for quality control of VD3-treated regulatory DCs in future clinical studies.

The aim of the study was to investigate clinical and microbial effects of probiotic candidate strains in patients with moderate gingivitis. The null hypothesis was that the clinical measurements with treatment would not differ from placebo. 47 adult patients were enrolled in a randomised placebo-controlled trial with a 4-week intervention of tablets containing a mix of Lactobacillus rhamnosus PB01, DSM 14869 and Lactobacillus curvatus EB10, DSM 32307 or placebo. Clinical examinations and samplings were done at baseline and after 2, 4 and 6 weeks. The clinical endpoints were general bleeding on probing (BOP), general plaque index (PI) and flow of gingival crevicular fluid (GCF). In addition, the concentration of selected cytokines (interleukin (IL)-1β, IL-6, IL-8, IL-10, tumour necrosis factor alpha (TNF-α)) in GCF was determined with multiplex immunoassays. The profiles of the salivary microbiome were analysed with Next Generation Sequencing (NGS) and qPCR. In contrast to the placebo group, there was a significant reduction in BOP and amount of GCF (P<0.05) after 4 weeks in the probiotic test group when compared with baseline. The general PI was less affected although there was a tendency of decreased plaque levels in the probiotic group (P=0.05-0.09). The cytokines were unaffected by the intervention as well as the salivary microbiome. The Shannon index showed no significant differences between the groups or alterations over time. The occurrence of both probiotic strains increased in saliva of the test subjects during the intervention but returned to baseline levels within 2 weeks. Although a marked improvement in gingival health was recorded in the probiotic group, the null hypothesis could not be rejected.