Antibiotic resistance in bacteria and its future for novel antibiotic development

Since the first introduction of the sulfa drugs and penicillin into clinical use, large numbers of antibiotics have been developed and hence contributed to human health. But extensive use of antibiotics has raised a serious public health problem due to multiantibiotic resistant bacterial pathogens that inevitably develop resistance to every new drug launched in the clinic. Consequently, there is a pressing need to develop new antibiotics to keep pace with bacterial resistance. Recent advances in microbial genomics and X-ray crystallography provide opportunities to identify novel antibacterial targets for the development of new classes of antibiotics and to design more potent antimicrobial compounds derived from existing antibiotics respectively. To prevent and control infectious diseases caused by multiantibiotic resistant bacteria, we need to understand more about the molecular aspects of the pathogens' physiology and to pursue ways to prolong the life of precious antibiotics.     

A chloride-inducible acid resistance mechanism in Lactococcus lactis and its regulation

Previously, a promoter was identified in Lactococcus lactis that is specifically induced by chloride. Here, we describe the nucleotide sequence and functional analysis of two genes transcribed from this promoter, gadC and gadB . GadC is homologous to putative glutamate-γ-aminobutyrate antiporters of Escherichia coli and Shigella flexneri and contains 12 putative membrane-spanning domains. GadB shows similarity to glutamate decarboxylases. A L . lactis gadB mutant and a strain that is unable to express both gadB and gadC was more sensitive to low pH than the wild type when NaCl and glutamate were present. Expression of gadCB in L . lactis in the presence of chloride was increased when the culture pH was allowed to decrease to low levels by omitting buffer from the medium, while glutamate also stimulated gadCB expression. Apparently, these genes encode a glutamate-dependent acid resistance mechanism of L . lactis that is optimally active under conditions in which it is needed to maintain viability. Immediately upstream of the chloride-dependent gadCB promoter P _gad , a third gene encodes a protein (GadR) that is homologous to the activator Rgg from Streptococcus gordonii . gadR expression is chloride and glutamate independent. A gadR mutant did not produce the 3 kb gadCB mRNA that is found in wild-type cells in the presence of NaCl, indicating that GadR is an activator of the gadCB operon.     

新型粒子群优化算法在多序列联配中的应用

将粒子群优化算法应用于序列联配,提出了一种改进的粒子群优化算法,该算法在粒子群的进化过程中根据粒子的适应值动态地调整粒子群的惯性权重与粒子群飞行速度范围,提高了算法的收敛速度和收敛精度;针对PSO算法可能出现的早熟现象,引入重新初始化机制,增强了算法的搜索能力,实验表明该算法是有效的。     

'DNA' as contaminants in antibiotics and its capacity to transform bacteria to drug resistance

DNA/and deoxyribose sugars were detected in streptomycin (Sm), kanamycin, polymyxin, penicillin G, ampicillin, methicillin, cloxacillin and mitomycin C in small amounts/traces. Stained DNA could be feebly visualized directly in Sm run in agarose gel, which improved after its separation and concentration. These DNA materials were DNase sensitive, RNase and pronase resistant, and appeared to consist of fragments, c. less than or equal to 6 Mdal; this could repeatedly transform to SmR several recipient enterobacteria and vibrios; E. coli C600 and S. typhi 57, after such transformation revealed similar plasmid DNA bands that were absent in their wild-types. G + C mole% of plasmid and chromosomal DNA of recipient (57) along with that of Streptomyces griseus reference strain, suggested an extraneous origin for the plasmid DNA.     

植物几丁质酶及其在抗真菌病害中的应用

植物几丁质酶的研究是抗真菌基因工程的热点之一。几丁质酶能够水解真菌细胞壁的主要成分几丁质,在植物抗真菌病害反应中发挥重要的作用。介绍了几丁质酶的基本生物学特性、基因的诱导表达,并对植物几丁质酶基因在抗真菌病害基因工程中的应用进行了阐述。     

A novel role for cholecystokinin: regulation of mesenteric vascular resistance

The aim of this work was to characterize the vasoactive effect of cholecystokinin on mesenteric vasculature. The mesenteric vascular bed of 3-month-old Sprague-Dawley rats was isolated and perfused at constant flow and changes in perfusion pressure monitored. CCK peptides lacked any direct contractile or relaxing effect on the mesenteric smooth muscle. Transmural nerve stimulation (TNS, 200 mA, 0.2 ms, 8 and 16 Hz) elicited an increase in perfusion pressure reflecting contraction of the bed and CCK inhibited neurogenic contractions elicited by 8 and 16 Hz TNS. The inhibition of neurogenic contractions was blocked by the CCK2 receptor (CCK2R) antagonist, L-365,260 (10 and 100 nM), but not by the CCK1R antagonist, SR-27897. The inhibition of neurogenic contractions was reversed by the non-specific NOS inhibitor, L-NAME as well as by the specific nNOS inhibitor, S-methyl-L-thiocitrulline. In whole-mount segments of mesenteric arteries, CCK2R was detected in the adventitia, in nerve terminals, where it co-localized with synaptophysin and nNOS. CCK-8 immunoreactive fibers were also detected. These results suggest that CCK mediates vasodilatation of the mesenteric vascular bed through the release of NO via its presynaptic CCK2R. Our findings provide, for the first time, a neural mechanism by which CCK may increase mesenteric blood flow.     

A novel insulin receptor-signaling platform and its link to insulin resistance and type 2 diabetes

Insulin-induced insulin receptor (IR) tyrosine kinase activation and insulin cell survival responses have been reported to be under the regulation of a membrane associated mammalian neuraminidase-1 (Neu1). The molecular mechanism(s) behind this process is unknown. Here, we uncover a novel Neu1 and matrix metalloproteinase-9 (MMP-9) cross-talk in alliance with neuromedin B G-protein coupled receptor (GPCR), which is essential for insulin-induced IR activation and cellular signaling. Neu1, MMP-9 and neuromedin B GPCR form a complex with IRβ subunit on the cell surface. Oseltamivir phosphate (Tamiflu®), anti-Neu1 antibodies, broad range MMP inhibitors piperazine and galardin (GM6001), MMP-9 specific inhibitor (MMP-9i), and GPCR neuromedin B specific antagonist BIM-23127 dose-dependently inhibited Neu1 activity associated with insulin stimulated rat hepatoma cells (HTCs) that overly express human IRs (HTC-IR). Tamiflu, anti-Neu1 antibodies and MMP-9i attenuated phosphorylation of IRβ and insulin receptor substrate-1 (IRS1) associated with insulin-stimulated cells. Olanzapine, an antipsychotic agent associated with insulin resistance, induced Neu3 sialidase activity in WG544 or 1140F01 human sialidosis fibroblast cells genetically defective in Neu1. Neu3 antagonist 2-deoxy-2,3-didehydro-N-acetylneuraminic acid (DANA) and anti-Neu3 antibodies inhibited sialidase activity associated with olanzapine treated murine Neu4 knockout macrophage cells. Olanzapine attenuated phosphorylation of IGF-R and IRS1 associated with insulin-stimulated human wild-type fibroblast cells. Our findings identify a novel insulin receptor-signaling platform that is critically essential for insulin-induced IRβ tyrosine kinase activation and cellular signaling. Olanzapine-induced Neu3 sialidase activity attenuated insulin-induced IGF-R and IRS1 phosphorylation contributing to insulin resistance.     

Inhibition of efflux pumps as a novel approach to combat drug resistance in bacteria

Efflux mechanisms have become broadly recognized as major components of resistance to many classes of antibiotics. Some efflux pumps selectively extrude specific antibiotics, while others, referred to as multidrug resistance (MDR) pumps, expel a variety of structurally diverse compounds with differing antibacterial modes of action. There are numerous potentially beneficial consequences of the inhibition of efflux pumps in improving the clinical performance of various antibiotics, and several companies and research laboratories have initiated programs to discover and develop efflux pump inhibitors. This review will summarize recent achievements in this new, very exciting and equally challenging field.     

A novel method for quantifying overdispersion in count data and its application to farmland birds

The statistical modelling of count data permeates the discipline of ecology. Such data often exhibit overdispersion compared with a standard Poisson distribution, so that the variance of the counts is greater than that of the mean. Whereas modelling to reveal the effects of explanatory variables on the mean is commonplace, overdispersion is generally regarded as a nuisance parameter to be accounted for and subsequently ignored. Instead, we propose a method that models the overdispersion as a biologically interesting property of a data set and show how novel inference is provided as a result. We adapted the double hierarchical generalized linear model approach to create an easily extendible model structure that quantifies the influence of explanatory variables on the overdispersion of count data, and apply it to farmland birds. These data were from a study within Irish agricultural ecosystems, in which total bird species abundance and the abundance of farmland indicator species were compared on dairy and non‐dairy farms in the winter and breeding seasons. In general, overdispersion in bird counts was greater on dairy farms than on non‐dairy farms, and for total bird numbers, overdispersion was greatest on dairy farms in winter. Our code is fitted using the Bayesian package Rstan, and we make all code and data available in a GitHub repository. Within a Bayesian framework, this approach facilitates a meaningful quantification of the effects of categorical explanatory variables on any response variable with a tendency to overdispersion that has a meaningful biological or ecological explanation.     

细菌锌离子调控体系与感染北大核心CSCD

锌(Zn)是生命体不可或缺的微量元素,对细菌和宿主同等重要。细菌体内锌离子稳态的维持依赖锌离子转运和调控体系。宿主通过限制锌离子或高锌离子中毒来控制细菌感染。为了在宿主体内生存,细菌必须表达高亲和力的锌离子转运系统,如ZnuABC,以获取足够的锌离子。由于锌与细菌大量的代谢和毒性通路密切相关,在细菌建立感染的过程中尤为重要,因此通过抑制锌离子转运系统来影响锌离子的稳态,将成为一个非常有发展前途的新型抗菌策略。     

一种挖掘关键基因的新方法及其应用(英文)

近来,人们发现从疾病相关基因中寻找关键基因对疾病的诊断和治疗很重要。癌相关基因的网络是根据正常和患病的胶质瘤组织的基因表达谱建立。根据建立的基因网络和CIPHER方法,不同阈值下的正常和患病的胶质瘤表型网络被建立。根据已知的疾病和表型间的关联,另一组正常和患病的胶质瘤表型网络被建立。将两种方法建立的相应的表型网络进行比较,匹配度最大时对应的阈值及基因和表型网络被确定。在此基础上,通过打分方法得到了7个关键基因:DMBT1,ERBB2,NF2,PDGFB,AR,ARAF和TP53。文献查询发现其中5个基因与胶质瘤的形成和发展密切相关。剩下两个基因中的ARAF也间接地参与胶质瘤形成。因此,这两个基因可能在胶质瘤的形成中起重要作用。这一预测仍需要实验验证。     

A new strategy to produce active human Src from bacteria for biochemical study of its regulation

Enzymological studies of Src protein tyrosine kinase have been hindered by the lack of a suitable bacterial expression system. Poor expression of active Src appears to be due to toxicity associated with its kinase activity. To overcome this problem, we fused Src to a protein tyrosine phosphatase with an affinity tag and an appropriate thrombin cleavage site. Upon affinity purification of the fusion protein, Src was released by thrombin digestion and further purified by FPLC. This strategy has been used to produce several Src mutants that display catalytic and regulatory properties similar to those from eukaryotic expression systems. Characterization of the Src mutants confirmed that inactivation of Src by Csk through tail tyrosine phosphorylation required the Src SH3 domain.