Role of Cys221 and Asn116 in the zinc-binding sites of the Aeromonas hydrophila metallo-β-lactamase

The CphA metallo--lactamase produced by Aeromonas hydrophila exhibits two zinc-binding sites. Maximum activity is obtained upon binding of one zinc ion, whereas binding of the second zinc ion results in a drastic decrease in the hydrolytic activity. In this study, we analyzed the role of Asn116 and Cys221, two residues of the active site. These residues were replaced by site-directed mutagenesis and the different mutants were characterized. The C221S and C221A mutants were seriously impaired in their ability to bind the first, catalytic zinc ion and were nearly completely inactive, indicating a major role for Cys221 in the binding of the catalytic metal ion. By contrast, the binding of the second zinc ion was only slightly affected, at least for the C221S mutant. Mutation of Asn116 did not lead to a drastic decrease in the hydrolytic activity, indicating that this residue does not play a key role in the catalytic mechanism. However, the substitution of Asn116 by a Cys or His residue resulted in an approximately fivefold increase in the affinity for the second, inhibitory zinc ion. Together, these data suggested that the first zinc ion is located in the binding site involving the Cys221 and that the second zinc ion binds in the binding site involving Asn116 and, presumably, His118 and His196.Received 3 March 2003; received after revision 4 August 2003; accepted 25 August 2003     

Pathogenic enterococci: new developments in the 21st century

Enterococci, traditionally viewed as Gram-positive commensal bacteria inhabiting the alimentary canals of humans and animals, are now acknowledged to be organisms capable of causing life-threatening infections in humans, especially in the nosocomial environment. The existence of enterococci in such a dual role is facilitated, at least in part, by its intrinsic and acquired resistance to virtually all antibiotics currently in use. Beginning with the initial identification of a streptococci of fecal origin in the late 19th century, enterococci have been studied for over a century now. A number of comprehensive reviews during this time have addressed various aspects of enterococci, including classification, biology, virulence, antibiotic resistance and so on. This review specifically addresses the important advances in the field of enterococcal research that have occurred since the beginning of the 21st century. Most notable among these developments have been the insights into enterococcal genomes and pathogenicity.Received 10 April 2003; received after revision 31 May 2003; accepted 3 June 2003     

电阻抗法在细菌耐药性试验中的应用

将电阻抗法应用于医学临床细菌耐药性试验。笔者用自制的微生物电阻抗传感器和电阻抗测量装量,通过测量接种细菌的培养基电阻抗值(V值),并将细菌在培养期间内取得的所有V值描记成“阻抗曲线图谱”,因细菌培养基所产生的阻抗曲线图谱与在细菌培养基中滴加了抗菌药物所形成的阻抗曲线图谱形态有明显差别,利用这种曲线形态的差别尝试用来判断细菌的耐药性、该研究对实现医学临床快速耐药性试验,药敏检测自动化、智能化有一定的实际意义．     

Studies on Wear and corrosion resistances of carbon nitride thin films on Ti alloy

0　IntroductionArtificialarthrosiseshavebeenwidelyusedintheoperatorsclinicalsurgeriesforthepatientswhosearthrosiseswerese riouslyseverelydamaged.Butmostofartificialarthrosisescanon lybeusedforabouttenyearslongduetotheseriousabrasion .Forthisreason ,theimprovementofthelongevityofartificialarthrosisbecomesanimportantsubjectofstudy .TheartificialarthrosisesbeingmadeupofTialloyTC4(Ti 6Al 4V)andthepolythenewithultrahighmolecularweighthavegreatpotentialitiesonim provinglongevitybecauseTC4canreac…     

混凝土施工缝留置位置与处理

砼施工缝留置原则运用于结构受剪力较小且便于施工的部位,但在施工中技术人员应根据砼构件的受力分析,重点结合《规范》条款规定的构件位置正确理解留置施工缝,并检测施工缝处砼强度后继续浇筑后续砼。     

ICP-AES比较临床分离药物敏感和耐药结核分枝杆菌的金属离子

利用现在多元素痕量分析最重要的方法——电感耦合等离子体原子发射光谱(ICP—AES)分析结核分枝杆菌中的金属离子的种类和含量,结果表明,Cr^3 ,Al^3 ,Ca^2 ,Fe^2 ,Mn^2 ,Co^2 ,Cu^2 ,Ni^2 的含量较高,并发现药物敏感菌株和耐药菌株之间在金属离子种类和含量上有明显区别．这种差异在耐药中的意义有待进一步研究．     

QTL mapping of the root traits and their correlation analysis with drought resistance using DH lines from paddy and upland rice cross

A Double Haploid (DH) population, 116 plants, derived from the cross between Japonica upland rice IRAT109 and paddy rice Yuefu, planted in PVC pipe under upland ecosystem in 2001 and 2002, was used in this study. Seven root traits, including basal root thickness (BRT), total root number (RN), maximum root length (MRL), root fresh weight (RFW), root dry weight (RDW), ratio of root fresh weight to shoot fresh weight (RFW/SFW) and ratio of root dry weight to shoot dry weight (RDW/SDW), were studied. Using index of drought resistance (IDR), the ratio of yield under upland ecosystem to yield under lowland ecosystem of DH lines, as the criteria of drought resistance, and correla-tion analysis between root traits and IDR, showed that BRT, MRL and RN were significantly correlated with IDR. High IDR lines had thicker BRT, longer MRL and less RN than low IDR lines. A molecular linkage map with 94 RFLP markers and 71 SSR markers covering 1535.1 cM was pro-duced. QTLs and G譋 interactions for BRT, RN, MRL, RFW, RDW, RFW/SFW and RDW/SDW were obtained based on the constructed molecular linkage map and soft-ware QTLmapper version 1.0. A total of 18 additive QTLs and 18 pairs of epistatic QTLs associated with root traits were detected. There were nine additive QTLs and two pairs of epistatic QTLs performed significant interactions with environment. Some QTLs with high general contribution and no G譋 interaction were obtained. Two pairs of epistatic QTLs mrl3 and mrl8, brt3 and brt11a controlling MRL and BRT had high general contributions of 21.51% and 13.03% respectively. An additive QTL and a pair of epistatic QTLs controlling RFW and RDW had high general contributions of 13.50% and 25.64% respectively. Marker assisted selec-tion (MAS) for rice drought resistance based on QTL with high general contribution, low G譋 interaction and tightly linkage with IDR were also discussed.     

轴承硬度与耐磨性关系的探讨

对轴承硬度与耐磨性进行检测.结果表明,轴承零件的硬度值只要控制在HRC61以上,就能保证获得良好的耐磨性.     

基于U-I模型的感应电机定子磁链估计的仿真研究

定子磁场定向的直接转矩控制逐渐被广泛应用于高性能异步电机调速系统。定子磁链估计精度是影响控制系统动态、稳态性能的主要因素。由于定子电阻参数变化及低通滤波器的影响，尤其低速运行时，传统定子磁链估计方法的估计精度及快速性往往很差。提出了一个全新的改进U-I模型定子磁链估计方案，不仅解决了定子电阻整定补偿问题。而且解决了低通滤波器截止频率优化补偿问题．在此基础上，建立直接转矩控制方案的仿真系统，仿真结果表明提出的方案能够使异步电机在低速到高速范围内都能得到令人满意的性能。     

一种新的变压器励磁涌流抑制技术

在探讨变压器励磁涌流的产生原理及其影响因素的基础上,分析现有励磁涌流抑制技术的优缺点,提出了一种抑制变压器励磁涌流的新技术.该技术将选相位关合法和串联电阻法相结合,考虑了剩磁的影响,在应用选相位关合法的基础上,通过设置适当的电阻及其切除时间来进一步降低励磁涌流.仿真实验表明,该技术能有效降低变压器空载合闸时的励磁涌流.