Biohydrogenation of linoleic acid by Clostridium sporogenes, Clostridium bifermentans, Clostridium sordellii and Bacteroides sp.

Abstract Several strains of Clostridium bifermentans, Clostridium sporogenes and Clostridium sordellit and one strain of Bacteroides sp. hydrogenate linoleic acid into transvaccenic acid in vitro following the same pathway. Linoleic acid (18:2; 9- cis , 12- cis ) was first isomerised into 9- cis , 11- trans -octadecadienoic acid, after which the 9- cis double bond was reduced. These species also hydrogenated linoleic acid into an octadecenoic acid in vivo when mono-associated with gnotobiotic rats. Several other species of Clostridium and Bacteroides did not hydrogenate linoleic acid.     

Detection of sickle-cell mutation by electrophoresis of partial RNA:DNA hybrids following solution hybridization

We have developed a method in which partially single-stranded (ss) DNA molecules containing a defined region of duplex RNA:DNA are electrophoretically separated in agarose gels. The partial hybrids are formed by solution hybridization with a uniform length RNA probe complementary to part of the DNA sequence of interest. Following hybridization, the RNA/DNA mixture is fractionated by agarose gel electrophoresis at high temperature to minimize intrastrand base pairing which causes mobility heterogeneity. Not requiring the steps of DNA transfer from the gel to a solid support and subsequent probing, pre-electrophoretic hybridization allows the direct identification of single-copy fragments. Conditions for the detection of single-copy genes in human DNA digested with specific restriction endonucleases were developed and applied to the diagnosis of sickle-cell disease. This method should be applicable for the analysis of DNAs of high complexity where the presence of DNA polymorphisms and interspersed repeated DNA sequences often make impossible the creation of complete RNA:DNA hybrids.     

Structural basis for the flexible recognition of α‐glucan substrates by Bacteroides thetaiotaomicron SusG

Bacteria that reside in the mammalian intestinal tract efficiently hydrolyze dietary carbohydrates, including starch, that escape digestion in the small intestine. Starch is an abundant dietary carbohydrate comprised of α1,4 and α1,6 linked glucose, yet mammalian intestinal glucoamylases cannot effectively hydrolyze starch that has frequent α1,6 branching as these structures hinder recognition and processing by α1,4‐specific amylases. Here we present the structure of the cell surface amylase SusG from Bacteroides thetaiotaomicron complexed with a mixed linkage amylosaccharide generated from transglycosylation during crystallization. Although SusG is specific for α1,4 glucosidic bonds, binding of this new oligosaccharide at the active site demonstrates that SusG can accommodate α1,6 branch points at subsite ?3 to ?2, and also at subsite+1 adjacent to the site of hydrolysis, explaining how this enzyme may be able to process a wide range of limit dextrins in the intestinal environment. These data suggest that B. thetaiotaomicron and related organisms may have a selective advantage for amylosaccharide scavenging in the gut.     

维吾尔族新发2型糖尿病患者肠道 乳杆菌属和多形拟杆菌的定量研究

目的定量研究维吾尔族新发2型糖尿病(T2DM)患者和糖耐量正常(NGT)人群肠道中乳杆菌属(Lactobacillus genus)和多形拟杆菌(Bacteroides thetaiotaomicron)的相对水平。方法严格按照纳入标准、排除标准收集维吾尔族新发T2DM患者96例,NGT 98例。提取所有研究对象的粪便细菌总DNA后,采用16SrDNA基因实时荧光定量PCR(Real-time PCR)对乳杆菌属和多形拟杆菌的水平进行定量检测;运用Pearson相关性分析乳杆菌属与研究对象的BMI、腰围、臀围、收缩压(SBP)、舒张压(DBP)、空腹血糖(FBG)、甘油三酯(TG)、总胆固醇(TC)、高密度脂蛋白(HDL-C)和低密度脂蛋白(LDL-C)的相关性。结果 16SrDNA基因Real-time PCR结果显示:(1)与新疆维吾尔族NGT组相比,乳杆菌属水平在新发T2DM中较低,差异有统计学意义(t=8.557,P=0.000)。但是多形拟杆菌在两组差异无统计学意义(t=0.524,P=0.601);(2)新疆维吾尔族上述人群肠道中乳杆菌属水平与FBG呈负相关(r=-0.334,P=0.000),腰围呈负相关(r=-0.170,P=0.018),TC呈负相关(r=-0.178,P=0.013),TG呈负相关(r=-0.157,P=0.030),收缩压呈负相关(r=-0.255,P=0.000)。结论 Lactobacillus genus水平在肠道中降低可能与2型糖尿病的发病,血糖和血脂代谢有关,其机制需进一步研究探讨。     

Single mid-chain GlcNAcβ1-6Galβ1-4R sequences of linear oligosaccharides are resistant to endo-β-galactosidase ofBacteroides fragilis

Endo--galactosidase (EC 3.2.1.103) ofBacteroides fragilis, at 250 mU ml^–1, did not cleave the internal galactosidic linkage of the linear radiolabelled trisaccharide GlcNAc1-6Gal1-4GlcNAc, or those of the tetrasaccharides Gal1-4GlcNAc1-6Gal1-4GlcNAc and Gal1-4GlcNAc1-6Gal1-4Glc. The isomeric glycans which contained the GlcNAc1-3Gal1-4GlcNAc/Glc sequence were readily cleaved.Abbreviations GlcNAc 2-acetamido-2-deoxy-d-glucose - Lact lactose - MT maltotriose - MTet maltotetraose - R _MTet chromatographic migration rate in relation to that of maltotetraose     

The use of bacteriophages of Bacteroides fragilis as indicators of the efficiency of virucidal products

The potential use of bacteriophage B40-8 of Bacteroides fragilis for the evaluation of the virucidal activity of antiseptics or disinfectants was investigated. The antiviral activity of two antiseptics and two disinfectants was evaluated according to a standard guideline. The effect of the virucidal agents was assessed on (i) viruses usually spread by direct contact with surfaces with contaminated secretions, i.e. herpes virus 1 and 2, and vaccinia virus, and (ii) viruses transmitted by the fecal-oral route, i.e. hepatitis A virus, poliovirus, adenovirus and rotavirus. The survival of B40-8 always equalled or exceeded that of the animal viruses tested. Our data suggest the use of bacteriophage B40-8 to complement the information furnished by some standardized methods in ascertaining the antiviral activity of virucidal preparations.     

分娩后宫颈分泌物中脆弱类杆菌的调查

本文介绍用脆弱类杆菌抗血清标记SpA(Bf—SpA)做协同凝集试验(CoA)调查123例正常分娩后妇女宫颈处的脆弱类杆菌,阳性率为22.7％,与常规厌氧培养法相比,SpA协同凝集试验具有特异性强、敏感性高、方法简便、快速的优点,适宜在基层单位推广应用。     

对脆弱类杆菌β-内酰胺酶及其与肠道微生态学关系的初步探讨

本文利用β-内酰胺酶作为分类学研究的方法,探讨了脆弱类杆菌与肠道菌之间的微生态学关系。临床分离菌株脆弱类杆菌55的β-内酰胺酶经离子交换层析、凝胶过滤和制备型聚丙烯酰胺凝胶电泳进行纯化,纯酶与脂质体-CPS-K佐剂混合制备抗血清,并建立IgG-ELISA和Western blotting方法,其测定结果均表明脆弱类杆菌的β-内酰胺酶不同于其它类杆菌和肠道菌的β-内酰胺酶,具有种的特异性。     

用间接免疫荧光法快速检测牙周袋内牙龈拟杆菌

我们用牙龈拟杆菌参考菌株(47A-1)制备标准免疫血清,通过问接免疫荧光法直接检测人龈下菌斑的涂片,共检查了90例正常人、75例牙龈炎患者、70例成年牙周炎患者。从记录一个荧光显微镜视野中的牙龈拟杆菌菌数发现:健康人与患者的标本间有一个明显的分界线,它具有帮助临床工作者诊断的实际意义。