Sequences related to the major subunit gene fedA of F107 fimbriae in porcine Escherichia coli strains that express adhesive fimbriae

Abstract Porcine Escherichia coli strains isolated from cases fo postweaning diarrhea or edema disease were analysed for the presence of fedA , the major subunit gene of F107 fimbriae. The E. coli isolates were known to contain colonisation factor '8813', or to express F107, 2134P or other fimbriae, different from F4, F5, F6, and F41. PCR with fedA -specific primers, restriction enzyme digestion of the PCR product, and nucleotide sequence analysis demonstrated that 2134P pili, colonisation factor '8813' and fimbriae identified on Australian strains of the O141 serotype belong to one family of F107 fimbrial antigens.     

Isolation of K88 Antigen Variants (ab,ac, ad) from Porcine Enterotoxigenic Escherichia coli Belonging to Different Serotypes

Fimbriae isolation by means of thermal shock was applied to fifteen K88-positive (three K88ab, nine K88ac and three K88ad) Escherichia coli reference strains belonging to serotypes O8:K87, O32, O45, O138:K81, O141:K85, O147:K89, O149:K91, and O157, as well as to ten K88-positive enterotoxigenic strains isolated from porcine diarrhea in Spain, all of them belonging to the O149 serogroup. Fimbriae were removed from the bacterial cells by thermal shock at 60 C and then precipitated using ammonium sulfate. The final amount of K88 antigen and the purification degree were not related to the serogroup of the bacteria or to the antigen variant but were related to the buffer used for isolation. Phosphate buffer containing urea was shown to be more effective than Tris-HCl for isolation of K88 antigen. The molecular weights by SDS-PAGE for K88ab, K88ac, and K88ad were 28.5, 29.2, and 31.0 kDa, respectively. All enterotoxigenic E. coli strains isolated in Spain showed the K88ac variant.     

Battling for Ribosomes: Translational Control at the Forefront of the Antiviral Response

In the early stages of infection, gaining control of the cellular protein synthesis machinery including its ribosomes is the ultimate combat objective for a virus. To successfully replicate, viruses unequivocally need to usurp and redeploy this machinery for translation of their own mRNA. In response, the host triggers global shutdown of translation while paradoxically allowing swift synthesis of antiviral proteins as a strategy to limit collateral damage. This fundamental conflict at the level of translational control defines the outcome of infection. As part of this special issue on molecular mechanisms of early virus–host cell interactions, we review the current state of knowledge regarding translational control during viral infection with specific emphasis on protein kinase RNA-activated and mammalian target of rapamycin-mediated mechanisms. We also describe recent technological advances that will allow unprecedented insight into how viruses and host cells battle for ribosomes.     

微生态制剂预防老年抗生素相关性腹泻的临床研究

目的 评价预防性应用微生态制剂(金双歧)在老年人抗生素相关性腹泻中的作用.方法 2010年5月至2013年2月应用抗生素的住院老年患者55例纳入观察对象,随机分为2组,预防组28例,给予微生态制剂双歧杆菌乳杆菌三联活菌片(金双歧)4片,3次/d,连续服用7～l0d;对照组27例,腹泻患者给予给予思密达3.76 g,3次/d.观察两组发生抗生素相关性腹泻情况,并进行肠道菌群分析.结果 预防组腹泻发生率、腹泻次数、腹泻持续时间与对照组比较,差异有统计学意义(P＜0.05).两组腹泻病例的大便培养及常规检查无异常,大便涂片检查出现细菌计数减少或大便杆球菌比失调;预防组粪检结果杆球比例倒置、细菌计数减少、真菌感染例数等与对照组比较差异有统计学意义(P＜0.05).结论 双歧杆菌乳杆菌三联活菌片(金双歧)可预防和减少老年人抗生素相关性腹泻的发生,减轻腹泻症状.     

家兔腹泻生态疗法的实验观察

根据动物微生态学理论,将3株正常菌：嗜酸乳杆菌、两歧双歧杆菌、粪链球菌研制成复方生态制剂,对家免细菌性腹泻进行治疗试验。共治疗75例,治愈率为96．00％,比痢特灵对照组治愈率提高16．00％。该制剂价格低廉,无毒、副作用,使用安全可靠,是一种防治家免细菌性腹泻的较理想制剂。     

猪流行性腹泻病毒S蛋白胞内区线性B细胞表位最小基序鉴定

【背景】S蛋白是猪流行性腹泻病毒(Porcine Epidemic Diarrhea Virus,PEDV)的主要结构蛋白和免疫原性蛋白,在前期的研究中,本课题组在S蛋白的胞内区鉴定到2个包含线性B细胞表位的短肽。【目的】鉴定PEDV S蛋白胞内区线性B细胞表位的最小基序。【方法】原核表达2个短肽的每次后移1个氨基酸的系列8肽,以兔抗S蛋白血清为一抗,通过Western Blot筛选阳性反应8肽,鉴定S蛋白胞内区线性B细胞表位的最小基序。【结果】S蛋白胞内区的2个包含线性B细胞表位的短肽共享一个表位,该表位的最小基序为¹³⁷¹QPYE¹³⁷⁴。同源性分析显示该B细胞表位基序为保守性表位。【结论】确定了S蛋白胞内区线性B细胞表位的最小基序为¹³⁷¹QPYE¹³⁷⁴;S蛋白抗原表位的鉴定有助于提高对其结构和功能的理解。     

三种检测婴幼儿腹泻A组轮状病毒的方法学评价

目的对婴幼儿腹泻A组轮状病毒的三种不同检测方法进行评价。方法同时采用胶体金、琼脂糖凝胶电泳、逆转录PCR（RT-PCR）对浙江萧山医院1周320份腹泻患儿的标本进行轮状病毒检测,然后进行方法学比较。结果三种方法的阳性检出率分别为36．3％、48．1％和42．8％,经卡方检验三种方法的检出率之间差异均存在统计学意义（P〈0．05）;以琼脂糖凝胶电泳为检测的最终标准,RT-PCR的敏感性为88．9％,特异性达100％;胶体金法的敏感性71．4％,特异性达96．4％。结论胶体金法检测轮状病毒的敏感性和特异性能够满足临床上对轮状病毒快速筛检的要求;PT-PCR法的敏感性和特异性均最高,既可用于临床检测,又是轮状病毒血清G、基因P分型的重要手段,在对无菌体液和环境样本中轮状病毒的检测具有不可比拟的优势。     

猪流行性腹泻病毒地方株LJB/03分离及培养特性

从黑龙江省某猪场疑为病毒性腹泻的发病猪采集腹泻粪便样品,以RT-PCR法扩增出猪流行性腹泻病毒M基因后,采用细胞培养法进行病毒分离。对细胞培养分离物进行间接免疫荧光、免疫电镜观察、RT-PCR及ELISA法检验,其中间接免疫荧光试验可见培养细胞中存在明显的特异性绿色荧光;免疫电镜下可见大小符合预期、有囊膜、花瓣状的典型冠状病毒结构特征;RT-PCR检测证实存在PEDVM基因;间接ELISA检测中平均P/N比值为7.6;从而确认为分离到一株猪流行性腹泻病毒(Porcine epidemic diarrhea virus,PEDV),命名为PEDVLJB/03株。随后,对该分离毒株的培养特性及如何提高病毒滴度进行探索。通过摸索该分离毒株的蚀斑形成条件,建立了PEDV蚀斑形成方法,并采用该方法进行病毒的蚀斑纯化,纯化得到PEDV大蚀斑克隆株和小蚀斑克隆株。对大、小两种蚀斑克隆株的病毒滴度测定结果表明,大小蚀斑克隆株细胞感染滴度相差明显。     

葛根芩连汤对泄泻肠道湿热证小鼠肠道双糖酶活性的影响

目的探究葛根芩连汤对泄泻肠道湿热证小鼠肠道双糖酶活性的影响。方法采用“高糖高脂+高温高湿+白酒复合灌胃冰水”的方法制备泄泻肠道湿热证小鼠模型,并运用葛根芩连汤进行干预治疗。分别在干预的第0、2、4、6天无菌采集各组小鼠肠黏膜,运用DNS法测定蔗糖酶活性,ONPG法测定乳糖酶活性,观察葛根芩连汤对泄泻肠道湿热证小鼠肠道双糖酶活性的影响。结果经肠道湿热泄泻造模后,小鼠肠道前段和后段黏膜的乳糖酶和蔗糖酶活性均显著下降,与正常组小鼠相比差异有统计学意义(均P<0.01)。随着治疗时间的延长,治疗组小鼠肠道乳糖酶和蔗糖酶活性与正常组相比有显著的提高(均P<0.05),治疗6 d后,肠道前段黏膜蔗糖酶和乳糖酶活性恢复至正常组水平(P>0.05)。肠道后段乳糖酶活性在第4天时最高,后段黏膜蔗糖酶活性在第6天时最高,与正常组和自愈组相比差异有统计学意义(均P<0.05)。结论泄泻肠道湿热证造模使小鼠肠道黏膜蔗糖酶、乳糖酶等双糖酶的活性显著下降,葛根芩连汤能调节泄泻肠道湿热证小鼠肠道黏膜乳糖酶和蔗糖酶的活性,从而发挥治疗泄泻肠道湿热证的疗效。     

Predictive markers of safety and immunogenicity of adjuvanted vaccines

Vaccination represents one of the greatest public health triumphs; in part due to the effect of adjuvants that have been included in vaccine preparations to boost the immune responses through different mechanisms. Although a variety of novel adjuvants have been under development, only a limited number have been approved by regulatory authorities for human vaccines. This report reflects the conclusions of a group of scientists from academia, regulatory agencies and industry who attended a conference on the current state of the art in the adjuvant field. Held at the U.S. Pharmacopeial Convention (USP) in Rockville, Maryland, USA, from 18 to 19 April 2013 and organized by the International Association for Biologicals (IABS), the conference focused particularly on the future development of effective adjuvants and adjuvanted vaccines and on overcoming major hurdles, such as safety and immunogenicity assessment, as well as regulatory scrutiny. More information on the conference output can be found on the IABS website, http://www.iabs.org/.