To determine if there is any change with age in the distribution of Bi dobacterium and Enterobacteriaceae species in human intestinal micro ora, strains were isolated from a total of 54 samples of human faeces (15 children, 3-15 years old; 17 adults, 30-46 years old; 22 elderly, 69-89 years old). The bi dobacteria were identi ed at species level using a numerical phenotypic identi cation system developed on the Internet (http:::kounou.lille.inra.fr) and DNA-DNA hyridization. The enterobacteria were identi ed using the API 20E system (Biomérieux, France). Escherichia coli was present in 93% of samples. Enterobacter and Klebsiella species were more frequently found in children while Proteus and Pro×idencia species were typically found in the elderly. The species Bi dobacterium longum was the most frequently species isolated in children and adults, whereas Bi dobacterium adolescentis was the most encountered species in the elderly. The distribution of B. adolescentis was signi cantly split between the children and the elderly age groups. There is a noticeable trend whereby Bi dobacterium pseudocatenulatu m occurs more frequently in children than in the elderly. The species Bi dobacterium infantis, Bi dobacterium gallicum and Bi dobacterium angulatum have never been isolated, while Bi dobacterium bre×e has only been isolated on one occasion.
Staphylococcus aureus gyrA and gyrB genes encoding DNA gyrase subunits were cloned and coexpressed in Escherichia coli under the control of the T7 promoter-T7 RNA polymerase system, leading to soluble gyrase which was purified to homogeneity. Purified gyrase was catalytically indistinguishable from the gyrase purified from S. aureus and did not contain detectable amounts of topoisomerases from the E. coli host. Topoisomerase IV subunits GrlA and GrlB from S. aureus were also expressed in E. coli and were separately purified to apparent homogeneity. Topoisomerase IV, which was reconstituted by mixing equimolar amounts of GrlA and GrlB, had both ATP-dependent decatenation and DNA relaxation activities in vitro. This enzyme was more sensitive than gyrase to inhibition by typical fluoroquinolone antimicrobial agents such as ciprofloxacin or sparfloxacin, adding strong support to genetic studies which indicate that topoisomerase IV is the primary target of fluoroquinolones in S. aureus. The results obtained with ofloxacin suggest that this fluoroquinolone could also primarily target gyrase. No cleavable complex could be detected with S. aureus gyrase upon incubation with ciprofloxacin or sparfloxacin at concentrations which fully inhibit DNA supercoiling. This suggests that these drugs do not stabilize the open DNA-gyrase complex, at least under standard in vitro incubation conditions, but are more likely to interfere primarily with the DNA breakage step, contrary to what has been reported with E. coli gyrase. Both S. aureus gyrase-catalyzed DNA supercoiling and S. aureus topoisomerase IV-catalyzed decatenation were dramatically stimulated by potassium glutamate or aspartate (500- and 50-fold by 700 and 350 mM glutamate, respectively), whereas topoisomerase IV-dependent DNA relaxation was inhibited 3-fold by 350 mM glutamate. The relevance of the effect of dicarboxylic amino acids on the activities of type II topoisomerases is discussed with regard to the intracellular osmolite composition of S. aureus.
Background: The consumption of dietary supplements (DS) has increased by almost 10% last decade. This trend may be true also for pregnant women; however, it was reported that pregnant women do not sufficient and correct knowledge on nutritional needs during pregnancy. Thus, we here attempted to determine the French situation of how French pregnant women take DS and its social-clinical significance using questionnaires, with special reference to vitamin D (VD) overtaking. Materials and Methods: A prospective multi-centric study was performed in three maternities in Paris. One hundred-fifty questionnaires were administered to pregnant women by midwife. Results: Thirty-four patients were interviewed at the maternity ward of the Deaconesses, 61 at the Pitié-Salpêtrière and 55 at Armand Trousseau. During pregnancy, DS started mainly during the first trimester. Frequently, doctors initially prescribe them and women tended to continue to take them thereafter. Possible VD overdosage was observed. Conclusion: We here characterized French pregnant women in terms of DS during pregnancy and also, to our knowledge, for the first time emphasized the risk of vitamin D overdose during pregnancy in France. However, since the cumulative dose throughout the pregnancy was not determined here, definite conclusion of whether women really take overdose of this supplement may not be made. Women of higher socio-professional society tended to consume more DS during pregnancy, and, thus, further monitoring may be needed especially to this fraction of pregnant women.
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