Hydrolysis of γ -glutamyl linkages by Fusobacterium nucleatum

The cell extracts of two human oral strains (FN2 and FN3) of Fusobacterium nucleatum displayed exceptionally high γ -glutamylpeptidase activity as determined with N-γ - l -glutamyl-2-naphthylamine as substrate. This activity was so dominant that the hydrolysis of other N -aminoacyl-2-naphthylamines progressed at a rate <10% of the former. Two major enzymes (I and II) were partially purified from FN2. I had a molecular weight of 115,000 and did not hydrolyze γ -glutamylcysteinylglycine (glutathione). II had a molecular weight of 70,000 and rapidly liberated only glutamic acid from glutathione. Strain FN3 contained several enzymes hydrolyzing γ -glu-2NA. Direct anion exchange chromatography of FN3 cell extracts separated one enzyme that liberated both glutamic acid and glycine from glutathione, one that was inactive against glutathione (but hydrolyzed γ -glu-2NA), and one that liberated only glutamic acid. Although γ -glu-2NA was a good synthetic substrate, glutathione was hydrolyzed at least 500 times faster by an enzyme present in both strains. These results indicate that the presence of γ -glutamylpeptidase activity is very characteristic of these F. nucleatum strains.Peer Reviewedhttp://deepblue.lib.umich.edu/bitstream/2027.42/41334/1/284_2005_Article_BF02094016.pd

Adverse maternal, fetal, and newborn outcomes among pregnant women with SARS-CoV-2 infection: an individual participant data meta-analysis.

INTRODUCTION: Despite a growing body of research on the risks of SARS-CoV-2 infection during pregnancy, there is continued controversy given heterogeneity in the quality and design of published studies. METHODS: We screened ongoing studies in our sequential, prospective meta-analysis. We pooled individual participant data to estimate the absolute and relative risk (RR) of adverse outcomes among pregnant women with SARS-CoV-2 infection, compared with confirmed negative pregnancies. We evaluated the risk of bias using a modified Newcastle-Ottawa Scale. RESULTS: We screened 137 studies and included 12 studies in 12 countries involving 13 136 pregnant women.Pregnant women with SARS-CoV-2 infection-as compared with uninfected pregnant women-were at significantly increased risk of maternal mortality (10 studies; n=1490; RR 7.68, 95% CI 1.70 to 34.61); admission to intensive care unit (8 studies; n=6660; RR 3.81, 95% CI 2.03 to 7.17); receiving mechanical ventilation (7 studies; n=4887; RR 15.23, 95% CI 4.32 to 53.71); receiving any critical care (7 studies; n=4735; RR 5.48, 95% CI 2.57 to 11.72); and being diagnosed with pneumonia (6 studies; n=4573; RR 23.46, 95% CI 3.03 to 181.39) and thromboembolic disease (8 studies; n=5146; RR 5.50, 95% CI 1.12 to 27.12).Neonates born to women with SARS-CoV-2 infection were more likely to be admitted to a neonatal care unit after birth (7 studies; n=7637; RR 1.86, 95% CI 1.12 to 3.08); be born preterm (7 studies; n=6233; RR 1.71, 95% CI 1.28 to 2.29) or moderately preterm (7 studies; n=6071; RR 2.92, 95% CI 1.88 to 4.54); and to be born low birth weight (12 studies; n=11 930; RR 1.19, 95% CI 1.02 to 1.40). Infection was not linked to stillbirth. Studies were generally at low or moderate risk of bias. CONCLUSIONS: This analysis indicates that SARS-CoV-2 infection at any time during pregnancy increases the risk of maternal death, severe maternal morbidities and neonatal morbidity, but not stillbirth or intrauterine growth restriction. As more data become available, we will update these findings per the published protocol

Acid production in dental plaque after exposure to probiotic bacteria

<p>Abstract</p> <p>Background</p> <p>The increasing interest in probiotic lactobacilli in health maintenance has raised the question of potential risks. One possible side effect could be an increased acidogenicity in dental plaque. The aim of this study was to investigate the effect of probiotic lactobacilli on plaque lactic acid (LA) production <it>in vitro</it> and <it>in vivo</it>.</p> <p>Methods</p> <p>In the first part (A), suspensions of two lactobacilli strains (<it>L. reuteri</it> DSM 17938<it>, L. plantarum</it> 299v) were added to suspensions of supragingival dental plaque collected from healthy young adults (n=25). LA production after fermentation with either xylitol or fructose was analyzed. In the second part (B), subjects (n=18) were given lozenges with probiotic lactobacilli (<it>L. reuteri</it> DSM 17938 and ATCC PTA 5289) or placebo for two weeks in a double-blinded, randomized cross-over trial. The concentration of LA in supragingival plaque samples was determined at baseline and after 2 weeks. Salivary counts of mutans streptococci (MS) and lactobacilli were estimated with chair-side methods.</p> <p>Results</p> <p>Plaque suspensions with <it>L. reuteri</it> DSM 17938 produced significantly less LA compared with <it>L. plantarum</it> 299v or controls (p<0.05). Fructose gave higher LA concentrations than xylitol. In part B, there were no significant differences in LA production between baseline and follow up in any of the groups and no differences between test and placebo were displayed. The salivary MS counts were not significantly altered during the intervention but the lactobacilli counts increased significantly in the test group (p<0.05).</p> <p>Conclusion</p> <p>Lactic acid production in suspensions of plaque and probiotic lactobacilli was strain-dependant and the present study provides no evidence of an increase in plaque acidity by the supply of selected probiotic lactobacilli when challenged by fructose or xylitol. The study protocol was approved by The Danish National Committee on Biomedical Research Ethics (protocol no H-2-2010-112).</p> <p>Trial registration</p> <p>NCT01700712</p

Metabolic activity of Streptococcus mutans biofilms and gene expression during exposure to xylitol and sucrose

The objective of the study was to analyse Streptococcus mutans biofilms grown under different dietary conditions by using multifaceted methodological approaches to gain deeper insight into the cariogenic impact of carbohydrates. S. mutans biofilms were generated during a period of 24 h in the following media: Schaedler broth as a control medium containing endogenous glucose, Schaedler broth with an additional 5% sucrose, and Schaedler broth supplemented with 1% xylitol. The confocal laser scanning microscopy (CLSM)-based analyses of the microbial vitality, respiratory activity (5-cyano-2,3-ditolyl tetrazolium chloride, CTC) and production of extracellular polysaccharides (EPS) were performed separately in the inner, middle and outer biofilm layers. In addition to the microbiological sample testing, the glucose/sucrose consumption of the biofilm bacteria was quantified, and the expression of glucosyltransferases and other biofilm-associated genes was investigated. Xylitol exposure did not inhibit the viability of S. mutans biofilms, as monitored by the following experimental parameters: culture growth, vitality, CTC activity and EPS production. However, xylitol exposure caused a difference in gene expression compared to the control. GtfC was upregulated only in the presence of xylitol. Under xylitol exposure, gtfB was upregulated by a factor of 6, while under sucrose exposure, it was upregulated by a factor of three. Compared with glucose and xylitol, sucrose increased cell vitality in all biofilm layers. In all nutrient media, the intrinsic glucose was almost completely consumed by the cells of the S. mutans biofilm within 24 h. After 24 h of biofilm formation, the multiparametric measurements showed that xylitol in the presence of glucose caused predominantly genotypic differences but did not induce metabolic differences compared to the control. Thus, the availability of dietary carbohydrates in either a pure or combined form seems to affect the cariogenic potential of S. mutans biofilms

Are we ready for caries prevention through bacteriotherapy?

Recent insights in medical science indicate that human biofilms play an important role in health and well-being, and have put microbiota modulation through bacteriotherapy into focus. In dentistry, bacterial interference with probiotic bacteria to support the stability and diversity of oral biofilms has gained similar interest. Investigations in vitro into metabolic activity, co-aggregation, growth inhibition, bacteriocin production, and adhesion have collectively suggested a potential role for probiotic lactobacilli and bifidobacteria to modulate the oral microbial ecology. Likewise, short-term clinical studies with intermediate microbial endpoints indicate that interference with caries-associated bacteria seems possible through probiotic dairy products, tablets, lozenges and chewing gum in various dose regimens. Few randomized controlled clinical trials with caries outcomes are available, but three studies with preschool children and the elderly have demonstrated preventive fractions between 21% and 75%, following regular intake of milk supplemented with probiotic lactobacilli. However, further large-scale trials with orally derived anti-caries candidates are needed before we can say that we are ready for bacteriotherapy as an adjunct to complement the existing evidence-based methods for preventing and controlling caries in daily practice