Health Benefit Characterization of Dominant Lactobacilli in Traditional Doogh

The aim of our study was the characterization and evaluation of isolated lactic acid bacteria(LAB

EGCG Prevents High Fat Diet-Induced Changes in Gut Microbiota, Decreases of DNA Strand Breaks, and Changes in Expression and DNA Methylation of Dnmt1

Obesity as a multifactorial disorder involves low-grade inflammation, increased reactive oxygen species incidence, gut microbiota aberrations, and epigenetic consequences. Thus, prevention and therapies with epigenetic active antioxidants, (-)-Epigallocatechin-3-gallate (EGCG), are of increasing interest. DNA damage, DNA methylation and gene expression of DNA methyltransferase 1, interleukin 6, and MutL homologue 1 were analyzed in C57BL/6J male mice fed a high-fat diet (HFD) or a control diet (CD) with and without EGCG supplementation. Gut microbiota was analyzed with quantitative real-time polymerase chain reaction. An induction of DNA damage was observed, as a consequence of HFD-feeding, whereas EGCG supplementation decreased DNA damage. HFD-feeding induced a higher inflammatory status. Supplementation reversed these effects, resulting in tissue specific gene expression and methylation patterns of DNA methyltransferase 1 and MutL homologue 1. HFD feeding caused a significant lower bacterial abundance. The Firmicutes/Bacteroidetes ratio is significantly lower in HFD + EGCG but higher in CD + EGCG compared to control groups. The results demonstrate the impact of EGCG on the one hand on gut microbiota which together with dietary components affects host health. On the other hand effects may derive from antioxidative activities as well as epigenetic modifications observed on CpG methylation but also likely to include other epigenetic elements

Changes in Human Fecal Microbiota Due to Chemotherapy Analyzed by TaqMan-PCR, 454 Sequencing and PCR-DGGE Fingerprinting

BACKGROUND: We investigated whether chemotherapy with the presence or absence of antibiotics against different kinds of cancer changed the gastrointestinal microbiota. METHODOLOGY/PRINCIPAL FINDINGS: Feces of 17 ambulant patients receiving chemotherapy with or without concomitant antibiotics were analyzed before and after the chemotherapy cycle at four time points in comparison to 17 gender-, age- and lifestyle-matched healthy controls. We targeted 16S rRNA genes of all bacteria, Bacteroides, bifidobacteria, Clostridium cluster IV and XIVa as well as C. difficile with TaqMan qPCR, denaturing gradient gel electrophoresis (DGGE) fingerprinting and high-throughput sequencing. After a significant drop in the abundance of microbiota (p = 0.037) following a single treatment the microbiota recovered within a few days. The chemotherapeutical treatment marginally affected the Bacteroides while the Clostridium cluster IV and XIVa were significantly more sensitive to chemotherapy and antibiotic treatment. DGGE fingerprinting showed decreased diversity of Clostridium cluster IV and XIVa in response to chemotherapy with cluster IV diversity being particularly affected by antibiotics. The occurrence of C. difficile in three out of seventeen subjects was accompanied by a decrease in the genera Bifidobacterium, Lactobacillus, Veillonella and Faecalibacterium prausnitzii. Enterococcus faecium increased following chemotherapy. CONCLUSIONS/SIGNIFICANCE: Despite high individual variations, these results suggest that the observed changes in the human gut microbiota may favor colonization with C. difficile and Enterococcus faecium. Perturbed microbiota may be a target for specific mitigation with safe pre- and probiotics

Vitamin E Modifies High-Fat Diet-Induced Increase of DNA Strand Breaks, and Changes in Expression and DNA Methylation of Dnmt1 and MLH1 in C57BL/6J Male Mice

Obesity is associated with low-grade inflammation, increased ROS production and DNA damage. Supplementation with antioxidants might ameliorate DNA damage and support epigenetic regulation of DNA repair. C57BL/6J male mice were fed a high-fat (HFD) or a control diet (CD) with and without vitamin E supplementation (4.5 mg/kg body weight (b.w.)) for four months. DNA damage, DNA promoter methylation and gene expression of Dnmt1 and a DNA repair gene (MLH1) were assayed in liver and colon. The HFD resulted in organ specific changes in DNA damage, the epigenetically important Dnmt1 gene, and the DNA repair gene MLH1. Vitamin E reduced DNA damage and showed organ-specific effects on MLH1 and Dnmt1 gene expression and methylation. These results suggest that interventions with antioxidants and epigenetic active food ingredients should be developed as an effective prevention for obesity—and oxidative stress—induced health risks

A PCR-DGGE fingerprinting of 16S rRNA coding regions of dominant bacteria over time.

<p>Bands that become stronger or nearly disappear following a single chemotherapeutic treatment are indicated with arrows. B Principal components analysis (PCA) based on dominant bacteria PCR-DGGE fingerprinting. The two outliers in the lower right corner of the plot are two samples of P07 following blood stem cell transplantation. C PCA illustrating the development of <i>Clostridium</i> cluster <i>IV</i> diversity in the course of chemotherapy and antibiotic treatment. Cluster <i>IV</i> diversity drops right after chemotherapy, causing a grouping of samples. Samples under antibiotic treatment (indicated as grey dots) group even closer, indicating a strong influence of antibiotics on <i>Clostridium</i> cluster <i>IV</i> diversity. A, sample of P01 before chemotherapy B, C and D, samples of P01 after chemotherapy; E, healthy control; SL, unrelated standard lane; black symbols… patients under chemotherapy and antibiotic treatment.</p

16S rRNA gene primers used for PCR-DGGE fingerprinting.

<p>16S rRNA gene primers used for PCR-DGGE fingerprinting.</p

TaqMan qPCR quantification of bacterial 16S rRNA coding regions showing lower abundance in patients undergoing chemotherapy and antibiotic treatment (P) than healthy controls (C).

<p>T₀, samples taken before a single shot of chemotherapy; T₁, 1–2 days after chemotherapy; T₂, 5–9 days after chemotherapy; Asterisk indicates a significant difference at p<0.05.</p

Heatmap showing abundances within the 454 sequencing dataset on the genus level.

<p>High throughput sequencing of samples P09 and P11 before (T₀) and after therapy (T₁) further helped to characterize the influence of a single chemotherapeutic cycle on the GI-microbiota. P11 was treated with chemotherapy alone and P09 also received antibiotic treatment.</p

Number of bands observed in PCR-DGGE fingerprinting in oncology patients before chemotherapy (T₀), immediately after chemotherapy (T₁) and 5–9 days after chemotherapy (T₂) and healthy controls averaged over all time points.

<p>Number of bands observed in PCR-DGGE fingerprinting in oncology patients before chemotherapy (T₀), immediately after chemotherapy (T₁) and 5–9 days after chemotherapy (T₂) and healthy controls averaged over all time points.</p

Phylogenetic tree showing the <i>Peptostreptococcaceae</i> found in samples from two oncology patients before and after chemotherapy.

<p>Identical sequences were grouped; the table on the right hand side shows their abundances in the 454 sequencing dataset. Sequences with >98.9% similarity to <i>Clostridium difficile</i> appeared only in samples taken immediately after chemotherapeutic cycles. Numbers indicate bootstrap values after 100 resamplings.</p