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Role of food in the stimulation of cytokine production.

To study the role of food in the stimulation of cytokine production, the effects of lactic bacteria on production of interferon alpha, beta, and gamma; interleukin 1 beta; and tumor necrosis factor alpha were evaluated in mice and humans. Yogurt bacteria induced plasma interferon alpha and beta production in mice. Yogurt intake containing 10(11) bacteria led to increased 2'-5' A synthetase activity in human blood mononuclear cells. This result may suggest an interferon action in a peripheral way. This effect was also found when subjects consumed 10(8) yogurt bacteria/d for 15 d. In an in vitro model, blood mononuclear cells cultured in the presence of yogurt bacteria produced interleukin 1 beta, tumor necrosis factor, and interferon alpha and gamma. These results suggest the involvement of a certain type of food in cytokine production under healthy conditions.

Adult↗

Novel disulphide esters of carbothioic acid as potent, non-detergent spermicides with low toxicity to Lactobacillus and HeLa cells in vitro.

BACKGROUND: The design, synthesis, characterization and evaluation of a novel series of non-detergent spermicides has led to the discovery of two unique molecules (DSE-36 and DSE-37) that were approximately 25 times more potent spermicides than nonoxynol-9 (N-9). METHODS: Normal human spermatozoa were used to assess the spermicidal activity (Sander-Cramer Assay), the effect on sperm-membrane integrity [hypo-osmotic swelling test (HOST)], supravital staining and scanning electron microscopy (SEM) and the induction of apoptosis [fluorescein isothiocyanate (FITC) Annexin-V and JC-1 labelling using flow cytometry] by the new class of compounds. HeLa and Lactobacillus cultures were used to assess the cytotoxicity of compounds and their compatibility to normal vaginal flora, respectively. RESULTS: Compounds DSE-36 and DSE-37 exhibited a strong spermicidal activity [minimum effective concentration (MEC) = 0.002%], which was approximately 25 times more potent than that of N-9 and Sapindus saponins (MEC = 0.05%). As compared with surfactants, DSE-36 and DSE-37 were found to be safer at MEC towards the growth and survival of Lactobacilli and HeLa cells in vitro and to have a milder effect on sperm plasma membrane. At EC(50) both induced apoptosis in sperm cells as characterized by increased labelling with Annexin-V and decreased polarization of sperm mitochondria. CONCLUSION: Preliminary studies have revealed that in sharp contrast to the non-specific surfactant action of N-9, DSE-36 and DSE-37 have a highly potent, mechanism-based, detrimental action on human sperm. The unique ability of these non-detergent molecules to selectively kill sperm and spare Lactobacilli and HeLa cells at MEC values much lower than that required for N-9 indicates their potential as superior ingredients for formulation into microbicidal contraceptives.

Cell Survival↗

Consumption of dairy produce and alcohol in a case-control study of breast cancer.

In a French case-control study of 1,010 breast cancer cases and 1,950 controls with nonmalignant diseases, the risk of breast cancer was found to be positively associated with frequency of cheese consumption and the level of fat in the milk consumed. A negative association was found between frequency of yogurt consumption and the risk of breast cancer. No association was found between the consumption of butter and the risk of breast cancer. The positive association between a daily consumption of alcohol and the risk of breast cancer previously reported was not altered when dairy produce consumption was taken into account.

Aged↗

Counteracting Fusarium proliferatum toxicity in broiler chicks by supplementing drinking water with Poultry Aid Plus.

To test whether Poultry Aid Plus (PAP, a commercial product for drinking water application) could reduce the stress on broiler chicks caused by Fusarium proliferatum contamination of feed, water (with or without PAP application, according to the manufacturer's instructions), and feed (experimentally infected with F. proliferatum fermented and dried corn culture material, CM) were provided to broiler chicks for 3 wk. Eight treatments consisting of a 2 (with or without PAP in water) x 4 (0, 1, 2, and 4% CM in feed) factorial design were tested in four replicate cages of six chicks each. The diet with 2% CM reduced weight gain by 23%; this reduction was preventable by PAP water application. The diet with 4% CM caused a cumulative mortality of 87.5%, which was reduced by PAP water application to 50%. The population half-life of the chicks on the diet with 4% CM was 6.5 d; this half-life was prolonged to at least 21 d by PAP water application. The PAP application also reduced the relative weight of the small intestine and promoted Lactobacillus colonization of the large intestine regardless of the level of CM in feed. Therefore, water application of PAP can be a prophylactic measure for F. proliferatum toxicity in poultry production.

Animals↗

Real-time PCR detection of lactic acid bacteria in cecal contents of eimeria tenella-lnfected broilers fed soybean oligosaccharides and soluble soybean polysaccharides.

This experiment was conducted to test whether dietary soybean meal oligosaccharides (SMO) and water-soluble polysaccharides (SMP) can assist broiler chickens in resisting Eimeria tenella, and to determine the survival of lactic acid bacteria in cecal contents postinfection. All birds received a soybean meal-free diet. The 6 experimental treatments were as follows: positive (COR) and negative (COW) control groups, 2 groups fed diets containing either 1% SMO or 0.5% SMP from 1 to 11 d of age; a vaccinated group (VAC), and an anticoccidial medicated group (ANT). Chickens of all treatments except COW were orally infected with 1000 sporulated oocysts of E. tenella on d 15. Fecal oocyst shedding was monitored per treatment group between d 5 and 13 postinfection. Lactic acid bacteria (LAB) in cecal contents were evaluated by a real-time PCR technique on d 7 postinfection. The results showed that the SMO and SMP groups had a lower number of oocysts per gram of feces during the monitoring period than the COR group. Threshold cycles were 22.21, 27.68, 13.99, 14.92, 12.97, and 14.85, for COW, COR, SMO, SMP, VAC, and ANT groups, respectively; specific PCR products were confirmed by the results of melting curve analysis and agarose gel electrophoresis. The results suggest that these LAB communities were promoted by SMO and SMP and have a competitive exclusion function when broiler chickens are infected with E. tenella.

Animal Feed↗

Favourable effect of an acidified milk (LC-1) on Helicobacter pylori gastritis in man.

OBJECTIVE: The supernatant of Lactobacillus johnsonii La1 culture was shown to be bactericidal and to have a partial, acid-independent suppressive effect on Helicobacter pylori in humans. The aim of the present study was to investigate the effect of L. johnsonii La1-acidified milk (LC-1) on H. pylori infection. DESIGN AND METHODS: Fifty-three volunteers infected with H. pylori as determined by positive 13C-urea breath test and positive serology were randomized to receive either LC-1 or a placebo 180 ml twice a day for 3 weeks. All subjects also received clarithromycin 500 mg bid during the last two weeks of acidified milk therapy. Oesophagogastroduodenoscopy and biopsies were performed at inclusion and repeated 4-8 weeks after the end of the treatment. H. pylori infection was confirmed by urease test and histology. H. pylori density and inflammation were scored using a modified Sydney classification. RESULTS: LC-1 ingestion induced a decrease in H. pylori density in the antrum (P= 0.02) and the corpus (P= 0.04). LC-1 also reduced inflammation and gastritis activity in the antrum (P= 0.02 and P= 0.01, respectively) and of activity in the corpus (P= 0.02). Clarithromycin eradicated H. pylori in 26% of the subjects; LC-1 did not improve the antibiotic effect. CONCLUSION: These results suggest that H. pylori infection and gastritis can be down-regulated by LC-1.

Animals↗

Characterization and production of amylovorin L471, a bacteriocin purified from Lactobacillus amylovorus DCE 471 by a novel three-step method.

The strongly hydrophobic bacteriocin amylovorin L471 from Lactobacillus amylovorus DCE 471 was isolated and purified to homogeneity from complex culture broth by a novel, rapid and simple three-step protocol including (i) ammonium sulphate precipitation, (ii) chloroform/methanol extraction/precipitation and (iii) reversed-phase HPLC, the only chromatographic step involved. The molecular mass of the peptide was determined to be 4876.9 Da by electrospray mass spectrometric analysis. N-terminal amino acid sequencing identified 35 amino acid residues as being identical to the N-terminal sequence of lactobin A, a bacteriocin from another L. amylovorus strain. These non-identical strains produce bacteriocins that display small differences in molecular mass and inhibitory spectrum. The amino acid sequence of amylovorin L471 shared significant homology with lactacin X, one of the two bactericidal peptides produced by Lactobacillus johnsonii VPI11088. A purified amylovorin L471 preparation permitted confirmation of the inhibitory spectrum previously established with a crude extract. It displayed a bactericidal mode of action on lactobacilli after an extremely rapid adsorption to the target cells. Two Listeria spp. were only weakly sensitive. Amylovorin L471 appears to be produced constitutively. Ethanol not only stimulated specific bacteriocin production but also prevented adsorption of the bacteriocin molecules to the producer cells upon prolonged fermentation. The latter result supports the hypothesis that the apparent inactivation of bacteriocin observed during the stationary phase of batch fermentations is due to adsorption.

Amino Acid Sequence↗

Biphasic kinetics of growth and bacteriocin production with Lactobacillus amylovorus DCE 471 occur under stress conditions.

Micro-organisms used during the production of fermented foods are subjected to several abiotic stresses. Microbial survival during these processes strongly depends on the ability of the cells to adapt and become more tolerant to the environmental conditions. Cultivation of Lactobacillus amylovorus DCE 471, a potential strain for use during type II sourdough fermentations, at low temperatures, unfavourable pH and high salt concentrations resulted in biphasic growth patterns. In addition, two separate bacteriocin peaks, as well as a dramatic change in cellular morphology, were observed. In general, an increase of the specific bacteriocin production occurred during the second growth phase. Finally, the observed sugar consumption profiles were affected by the applied fermentation temperature. Moreover, the highest bacteriocin activity occurred during maltose consumption at a low constant temperature of 28 degrees C and a constant pH of 5.4. Plate counts from both growth phases revealed the existence of two colony types. Irregular colonies were found to outnumber smoother colonies during the first growth phase, while the second growth phase was characterized by a greater number of smooth colonies. Electron microscopy was used to investigate the observed morphological switch at the single-cell level. Single, rod-shaped cells changed into elongated cells that grew in chains. Colony and cell morphology changes coincided with the biphasic growth pattern.

Bacterial Typing Techniques↗

Role of internalization in the pathogenicity of Shiga toxin-producing Escherichia coli infection in a gnotobiotic murine model.

We investigated the role of bacterial internalization in the killing caused by Shiga toxin-producing Escherichia coli (STEC) infection using a gnotobiotic murine model. A high number of internalized STEC was found in the colonic epithelial cells of STEC-infected mice by both an ex vivo assay and transmission electron microscopy. Most of these mice were killed within 10 days after infection. However, the implantation of lactic acid bacteria in such mice before infection markedly decreased the number of internalized STECs and also completely protected these hosts from killing by a STEC infection. The inhibition of such internalization by immunoglobulin also prevented the hosts from being killed. The Shiga toxin levels in these hosts indicated an inhibition of the penetration of Shiga toxins produced in the colon to the underlying tissue. These results suggested that the internalization plays an important role in the pathogenicity caused by STEC infection in a gnotobiotic murine model.

Animals↗