双歧杆菌及肠致病性大肠杆菌粘附的细胞膜通透性研究

采用乳酸脱氢酶释放法比较研究双歧杆菌１０２７株及肠致生大肠杆菌（ＥＰＥＣ）对体外肠上皮细胞Ｌｏｖｏ细胞株粘附的细胞膜通透性,探讨它们对肠上皮细胞的不同生物学交谈２。结果表明,双歧杆菌粘附Ｌｏｖｏ细胞后,宿主细胞释放ＬＤＨ远较ＥＰＥＣ粘附的效果低,提示双歧杆菌粘附对主细胞膜通生影响不大,而ＥＥＣ则可损伤宿主细胞膜而增加其通性。因此,双歧杆菌作为生理性细菌可与肠上皮细胞和谐共生,这与ＥＰＥＣ的粘附损伤     

沙门菌、大肠杆菌和金黄色葡萄球菌的多重PCR检测

根据沙门菌invA基因、大肠杆菌phoA基因和金黄色葡萄球菌nuc基因序列,设计3对特异性引物进行多重PCR并对反应条件进行优化。结果表明3对引物能特异地扩增出284bp、622bp、484bp的目的条带;最佳反应条件为沙门菌、大肠杆菌、金黄色葡萄球菌的引物浓度分别为40nmol／L、40nmol／L、80nmol／L,Mg^2＋浓度2．4mmol／L,dNTP浓度2001μmol／L,Taq DNA聚合酶1.5u,退火温度55．0℃-57．4℃之间;在此条件下多重PCR同时检测DNA的敏感性分别是10．2pg、10．2pg、102．0pg,检测时间4h。建立的多重PCR是一种敏感、特异、准确、快速的方法,为同时检测食品中沙门菌、大肠杆菌和金黄色葡萄球菌奠定了基础。     

AgNO3对大肠杆菌和金黄色葡萄球菌的抗菌作用及机制

以大肠杆菌和金黄色葡萄球菌为模式菌,对AgNO3的抗菌效果进行研究,并对其抗菌机制作初步探讨。AgNO3对大肠杆菌的抑制生长曲线表明:2.891 mg/L的AgNO3能够完全抑制106个/mL的大肠杆菌细胞生长,AgNO3使大肠杆菌和金黄色葡萄球菌的延滞期加长,并且浓度越高,延滞期越长。另外,AgNO3对大肠杆菌和金黄色葡萄球菌脱氢酶的活性有明显影响,随着AgNO3浓度的提高,脱氢酶的活性逐渐降低。AgNO3溶液作用于细菌后,细菌表面疏水性均有不同程度地下降,且浓度越大对其影响也越明显,大肠杆菌的下降程度要大于金黄色葡萄球菌。     

金黄色葡萄球菌spa基因在大肠杆菌中受多启动子调控

利用重组ＤＮＡ技术在大肠杆菌中克隆并表达了金黄色葡萄球菌Ａ蛋白ＳｐＡ。在Ｅ。ｃｏｌｉ ＲＲ１中ＳｐＡ的表达量随培养温度的升高而增加,４２℃的表达量是２３℃的６倍,但该在热休克应答缺陷的Ｅ．ｃｏｌｉ ＣＡＧ５９７及ＣＡＧ６２７中却消失。当删除ｓｐａ基因上游的一段序列后,仍有ＳｐＡ表达,但其表达量的热休克效应在Ｅ．ｃｏｌｉＲＲ１中也消失。这表明被删除的序列后,仍有ＳｐＡ表达,但其表达量的热休克效应在Ｅ     

金黄色葡萄球菌反向线性杂交探针检测方法的建立

目的建立一种检测金黄色葡萄球菌的简单、快速、灵敏、准确的方法。方法根据金黄色葡萄球菌的耐热核酸酶nuc基因,设计一对通用引物及两条特异性探针,用生物素标记通用引物的5＇端,将两条特异性探针固定于硝酸纤维膜上,使PCR产物与探针杂交。结果建立的反向线性杂交探针方法,其检测限为2 ng/μL,检测特异性和准确性均为100%。结论建立的反向线性杂交检测方法具有较高的敏感性和特异性,可用于实验动物金黄色葡萄球菌的快速检测。     

不同类型铜合金板的杀菌特性

【目的】微生物对可接触表面的污染给公共卫生带来了极大的威胁。利用具有杀菌特性的铜及铜合金代替不锈钢等制品,可以降低消毒剂的使用和细菌的传播。【方法】通过分析3株金黄色葡萄球菌和2株大肠杆菌在铜及铜合金平板上的存活时间,对不同类型铜合金的杀菌特性进行了探索。【结果】铜合金平板的杀菌能力与其铜含量成正比;铜合金对同属细菌的杀菌能力相近,对不同属细菌则有一定差异;铜合金的杀菌效率与细菌对Cu2+抗性没有直接联系;铜合金杀菌的效率与细菌的细胞壁结构可能有很大关联。【结论】铜及铜合金是较好的杀菌材料。     

沙门菌、大肠杆菌和金黄色葡萄球菌的多重PCR检测*

根据沙门菌invA基因、大肠杆菌phoA基因和金黄色葡萄球菌nuc基因序列,设计3对特异性引物进行多重PCR并对反应条件进行优化。结果表明3对引物能特异地扩增出284bp、622bp、484bp的目的条带;最佳反应条件为沙门菌、大肠杆菌、金黄色葡萄球菌的引物浓度分别为40nmol/L、40nmol/L、80nmol/L,Mg²⁺浓度2.4mmol/L,dNTP浓度200μmol/L,TaqDNA聚合酶1.5U,退火温度55.0℃~57.4℃之间;在此条件下多重PCR同时检测DNA的敏     

金黄色葡萄球菌的侵袭和侵袭后过程

金黄色葡萄球菌粘附宿主细胞后,与宿主细胞受体发生反应,激发宿主的跨膜信号传递和细胞骨架重排,从而进入细胞。进入宿主细胞的金黄色葡萄球菌可以在胞内存活,并且繁殖,胞内存活的金黄色葡萄球菌可诱导细胞发生凋亡。在金黄色葡萄球菌的侵袭和侵袭后过程中,毒力调节因子Agr和Sar起了调节作用。     

乳酸杆菌细胞裂解物对金黄色葡萄球菌、大肠埃希菌的抑制作用

目的探讨乳杆菌DM8909裂解物在体内外对金黄色葡萄球菌、大肠埃希菌的抑制作用。方法通过对乳杆菌超声波破碎制成裂解物,分别用乳杆菌裂解物原液、裂解物稀释液、发酵上清液、乳杆菌活菌制剂进行体内、体外实验,观察乳杆菌各成分对金黄色葡萄球菌、大肠埃希菌的抑制作用。结果德氏乳酸杆菌裂解物对金黄色葡萄球菌、大肠埃希菌的抑制作用与乳杆菌活菌制剂的抑制作用相近。结论德氏乳酸杆菌裂解物在体内外对金黄色葡萄球菌、大肠埃希菌均有较强的抑制作用。     

金黄色葡萄球菌疫苗的研究进展

金黄色葡萄球菌是引发医院内感染的主要因素之一,近年来由于其多种耐药菌株的出现,发病率迅速攀升,而且通常的抗生素治疗已不可靠。国内外研究者致力于寻找能够预防金黄色葡萄球菌感染的疫苗的有效靶点,如毒素、荚膜多糖、胞外基质结合蛋白、表面多糖、表面蛋白、毒力因子表达调控蛋白等,希望研制出能有效预防金黄色葡萄球菌感染的疫苗。本文主要介绍其近年来的相关研究。     

细菌在阴离子交换树脂上的吸附研究

报道了细菌 (E .coliK12和S .aureus)在阴离子交换树脂 (SuperQ和DEAE)上的静态吸附行为。采用静态法测定 30℃以及 37℃下E .coliK12和S .aureus在SuperQ和DEAE上的吸附等温线 ,分别用Freundlich和Langmuir方程拟合。实验表明 ,用Fre undlich方程拟合 ,得到较为满意的结果 ,其相关系数最大为 0 .995 ,最小为 0 .991;Freundlich等温线指数 1/n =0 .3～ 0 .4 ;而且在30℃下比在 37℃下更有利于吸附。同样采用静态法测定了吸附量随pH值的变化趋势。结果表明 ,对E .coliK12和S .aureus在SuperQ上的吸附 ,pH >6 .0～ 7.0的吸附量较pH2 .0～ 5 .0之间的多。     

细菌持留与抗生素表型耐药机制

持留菌是细菌群体中一小部分具有表型耐药的细菌。自1944年被发现后,近几十年来因其在慢性持续性感染和生物膜感染中的重要作用而得到越来越多的重视。已有的研究结果表明,细菌持留的机理复杂,涉及的相关信号通路有毒素-抗毒素系统、细胞能量代谢及蛋白核酸合成等生理状态的降低、DNA保护修复系统、蛋白酶系统、反式翻译、外排泵系统等。虽然不同细菌的持留机理有一定的相似性和保守性,但不同细菌的持留机制也存在差异,如毒素-抗毒素系统在大肠埃希菌(Escherichia coli)中的过度激活可导致持留菌增加,但在金黄色葡萄球菌(Staphylococcus aureus)中却并无相同作用。本文从持留菌的研究历史出发,综述了当前对革兰氏阴性菌和阳性菌的持留机制方面的研究进展,同时探讨了在持留菌相关感染疾病方面的治疗策略,以期为更好地解决持留菌带来的问题,缩短治疗时间提供新的思路。     

Effects of 50 Hz rotating magnetic field on the viability of Escherichia coli and Staphylococcus aureus

This study presents results of research on the influence of rotating magnetic field (RMF) of the induction of 30?mT and the frequency of 50?Hz on the growth dynamics and cell metabolic activity of E. coli and S. aureus, depending on the exposure time. The studies showed that the RMF caused an increase in the growth and cell metabolic activity of all the analyzed bacterial strains, especially in the time interval t?=?30 to 150?min. However, it was also found that the optical density and cell metabolic activity after exposition to RMF were significantly higher in S. aureus cultures. In turn, the study of growth dynamics, revealed a rapid and a significant decrease in these values from t?=?90?min) in the case of E. coli samples. The obtained results prove that RMF (B?=?30?mT, f?=?50?Hz) has a stimulatory effect on the growth and metabolic activity of E. coli and S. aureus. Furthermore, taking into account the time of exposure, stronger influence of RMF on the viability was observed in S. aureus cultures, which may indicate that this effect depends on the shape of the exposed cells.     

Transformation and expression of a staphylococcal plasmid in Escherichia coli

Abstract A multiple antibiotic-resistant Staphylococcus aureus , was found to possess three plasmid bands in agarose gel electrophoresis. A plasmid of approximately 4.3 kb (pMC790/2) was found to code for ampicillin and tetracycline resistance and to have one Eco RI site when transformed into S. aureus RN 4220. pMC790/2 in unmodified form was transformed into a recA⁻ E. coli at a frequency of 1.2×10⁴ transformants/μg of plasmid DNA. Plasmid (pMC790/2) replicated, maintained itself stably and expressed far better in the E. coli host than in S. aureus .     

Immunomodulating effects of Toxoplasma gondii occur at an early stage of infection

Abstract An antibody to a 60-kDa protein associated with membrane-bound ribosomes (MBRP) in Staphylococcus aureus was shown to cross-react with a 64-kDa protein in Bacillus subtilis . Evidence is presented suggesting that this Bacillus protein is identical to a 64-kDa protein, possibly involved in protein secretion, described by Horiuchi et al. [Proc. Natl. Acad. Sci. USA 80 (1983) 3287–3291). Both the 60-kDa staphylococcal protein and the 64-kDa Bacillus protein were precipitated as a complex with three other proteins when immunoprecipitated with the staphylococcal MBRP antibody.     

Fuctional organization of the outer membrane of escherichia coli: Phage and colicin receptors as components of iron uptake systems

The functional interaction of outer memberane proteins of E. coli can be studied using phage and colicin receptors which are essential components of penetration systems. The uptake of ferric iron in the form of the ferrichrome complex requires the ton A and ton B functions in the outer membrane of E. coli. The ton A gene product is the receptor protein for phage T5 and is required together with the ton B function by the phages T1 anf ?80 to infect cells and by colicin M and the antibiotic albomycin, a structural analogue of ferrichrome, to kill cells. The ton B function is necessary for the uptake of ferric iron complexed by citrate. Iron complexed by enterochelin is only transported in the presence of the ton B and feu functions. Cells which have lost the feu function are resistant to the colicins B, I or V while ton B mutants are resistant to all colicins. The interaction of the ton A, Ton B, and feu functions apparently permits quite different “substrates” to overcome the permeablility barrier of the outer membrane. It was shown for ferrichrome dependent iron uptake that the complexing agent was not altered and could be used repeatedly. Only very low amounts of ³H-labeled ferrichrome were found in the cell. It is possible that the iron is mobilized in the membrane and that desferriferrichrome is released into the medium without having entered the cytoplasm. Growth on ferrichrome as the sole iron source waw used to select revertants of T5 resistant ton A mutants. All revertants exhibited wild-type properties with the exception of partial revertants. In these 4 strains, as in the ton A mutants, the ton A protein was not detectable by SDS polyacrylamide gel electrophoreses of outer membranes. Albomycin resistant mutants were selected and shown to fall into 5 categories: (1) ton A; (2) ton B mutants; (3) mutants with no iron transport defects and normal ton A/ton B functions, which might be target site mutants; (4) mutants which were deficient in ferrichrome-mediated iron uptake but had normal ton A/ton B functions. We tentatively consider that the defect might be located in the active transport system of the cytoplasmic membrane; (5) a variety of mutants with the following general properties: most of them were resistant to colicin M, transported iron poorly, and, like ton B mutants, contained additional proteins in the outer membrane. The outer membrane protein patterns of wild-type and ton B mutant strains were compared by slab gel electrophoresis in an attempt to identify a ton B protein. It was observed that under most growth conditions, ton B mutants overproduced 3 proteins of molecular weights 74,000–83,000. In extracted, iron-deficient medium, both the wild-type and ton B mutant strains had similar large amounts of these proteins in their outer membranes. The appearance of these proteins was suppressed by excess iron in both wild-type and mutant. From this evidence it is apparent that the proteins appear as a response to low intracellular iron rather than being controlled by the ton B gene. The nature of these proteins and their possible role in iron transport is disussed.     

Understanding urinary conditioning film components on ureteral stents: profiling protein components and evaluating their role in bacterial colonization

Ureteral stents are fraught with problems. A conditioning film attaches to the stent surface within hours of implantation; however, differences between stent types and their role in promoting encrustation and bacterial adhesion and colonization remain to be elucidated. The present work shows that the most common components do not differ between stent types or patients with the same indwelling stent, and contain components that may drive stent encrustation. Furthermore, unlike what was previously thought, the presence of a conditioning film does not increase bacterial adhesion and colonization of stents by uropathogens. Genitourinary cytokeratins are implicated in playing a significant role in conditioning film formation. Overall, stent biomaterial design to date has been unsuccessful in discovering an ideal coating to prevent encrustation and bacterial adhesion. This current study elucidates a more global understanding of urinary conditioning film components. It also supports specific focus on the importance of physical characteristics of the stent and how they can prevent encrustation and bacterial adhesion.