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1.
m-AMSA, an established inhibitor of eukaryotic type II topoisomerases, exerts its cidal effect by binding to the enzyme–DNA complex thus inhibiting the DNA religation step. The molecule and its analogues have been successfully used as chemotherapeutic agents against different forms of cancer. After virtual screening using a homology model of the Mycobacterium tuberculosis topoisomerase I, we identified m-AMSA as a high scoring hit. We demonstrate that m-AMSA can inhibit the DNA relaxation activity of topoisomerase I from M. tuberculosis and Mycobacterium smegmatis. In a whole cell assay, m-AMSA inhibited the growth of both the mycobacteria.  相似文献   
2.
利用线性混合效应模型模拟杉木人工林枝条生物量   总被引:2,自引:0,他引:2  
基于福建省将乐林场45株人工杉木解析木的572组枝条生物量数据,采用线性混合效应模型方法,建立杉木人工林枝条总生物量和枝、叶生物量的预测模型,并利用独立样本数据对模型进行检验.结果表明: 线性混合效应模型比传统多元线性回归模型的拟合精度高.不同随机效应参数的组合,其混合模型的精度不同.考虑异方差结构的混合模型能够消除数据间的异方差性,其精度更高,其中,对于枝条总生物量和叶生物量模型,以指数函数作为异方差结构时的模型精度最高;对于枝生物量模型,以常数加幂函数作为异方差结构时的模型精度最高.模型检验结果表明:对于杉木人工林枝条生物量预测模型,考虑随机效应和异方差结构的线性混合模型的检验精度比传统多元线性回归模型的精度有明显提高.  相似文献   
3.
细胞因子是由免疫细胞和某些非免疫细胞经刺激而合成、分泌的一类具有广泛生物学活性的小分子蛋白质,其作为细胞间信号传递分子,主要参与调节免疫应答、免疫细胞分化发育、组织修复、介导炎症反应、刺激造血功能等。micro RNA(mi RNA)是存在于真核细胞内的一种非编码小RNA,可以调控基因转录后的表达,同时还可作为不同生理和病理状态的分子标记。许多研究表明,细胞因子相关基因的多态性与结核感染、肺结核发病易感性密切相关,而mi RNA在肺部疾病的正负调节功能与肺部疾病感染的发生、发展、转化与治疗有关。我们简要叙述了细胞因子、mi RNA与结核分枝杆菌感染三者之间的关联,以期有利于及时筛查潜伏结核感染和肺结核患者,降低结核感染率和发病率。  相似文献   
4.
In recent years, the bacterium responsible for tuberculosis has been increasing its resistance to antibiotics resulting in new multidrug resistant Mycobacterium tuberculosis (MR-TB) and extensively drug-resistant tuberculosis (XDR-TB). In this study we use several analytical techniques including NMR, FT-ICR, TOF-MS, LC–MS and UV/Vis to study the copper–capreomycin complex. The copper (II) cation is used as a carrier for the antibiotic capreomycin. Once this structure was studied using NMR, FT-ICR, and MALDI-TOF-MS, the NIH-NIAID tuberculosis cell line for several Tb strains (including antibiotic resistant strains) were tested against up to seven variations of the copper–capreomycin complex. Different variations of copper improved the efficacy of capreomycin against Tb up to 250 fold against drug resistant strains of Tb.  相似文献   
5.
Reports in recent years indicate that the increasing emergence of resistance to drugs be using to TB treatment. The resistance to them severely affects to options for effective treatment. The emergence of multidrug-resistant tuberculosis has increased interest in understanding the mechanism of drug resistance in M. tuberculosis and the development of new therapeutics, diagnostics and vaccines. In this study, a label-free quantitative proteomics approach has been used to analyze proteome of multidrug-resistant and susceptible clinical isolates of M. tuberculosis and identify differences in protein abundance between the two groups. With this approach, we were able to identify a total of 1,583 proteins. The majority of identified proteins have predicted roles in lipid metabolism, intermediary metabolism, cell wall and cell processes. Comparative analysis revealed that 68 proteins identified by at least two peptides showed significant differences of at least twofolds in relative abundance between two groups. In all protein differences, the increase of some considering proteins such as NADH dehydrogenase, probable aldehyde dehydrogenase, cyclopropane mycolic acid synthase 3, probable arabinosyltransferase A, putative lipoprotein, uncharacterized oxidoreductase and six membrane proteins in resistant isolates might be involved in the drug resistance and to be potential diagnostic protein targets. The decrease in abundance of proteins related to secretion system and immunogenicity (ESAT-6-like proteins, ESX-1 secretion system associated proteins, O-antigen export system and MPT63) in the multidrug-resistant strains can be a defensive mechanism undertaken by the resistant cell.

Electronic supplementary material

The online version of this article (doi:10.1007/s12088-015-0511-2) contains supplementary material, which is available to authorized users.  相似文献   
6.
The ability of Mycobacterium tuberculosis to resist intraphagosomal stresses, such as oxygen radicals and low pH, is critical for its persistence. Here, we show that a cytoplasmic redox sensor, WhiB3, and the major M. tuberculosis thiol, mycothiol (MSH), are required to resist acidic stress during infection. WhiB3 regulates the expression of genes involved in lipid anabolism, secretion, and redox metabolism, in response to acidic pH. Furthermore, inactivation of the MSH pathway subverted the expression of whiB3 along with other pH-specific genes in M. tuberculosis. Using a genetic biosensor of mycothiol redox potential (EMSH), we demonstrated that a modest decrease in phagosomal pH is sufficient to generate redox heterogeneity in EMSH of the M. tuberculosis population in a WhiB3-dependent manner. Data indicate that M. tuberculosis needs low pH as a signal to alter cytoplasmic EMSH, which activates WhiB3-mediated gene expression and acid resistance. Importantly, WhiB3 regulates intraphagosomal pH by down-regulating the expression of innate immune genes and blocking phagosomal maturation. We show that this block in phagosomal maturation is in part due to WhiB3-dependent production of polyketide lipids. Consistent with these observations, MtbΔwhiB3 displayed intramacrophage survival defect, which can be rescued bypharmacological inhibition of phagosomal acidification. Last, MtbΔwhiB3 displayed marked attenuation in the lungs of guinea pigs. Altogether, our study revealed an intimate link between vacuolar acidification, redox physiology, and virulence in M. tuberculosis and discovered WhiB3 as crucial mediator of phagosomal maturation arrest and acid resistance in M. tuberculosis.  相似文献   
7.
目的探讨结核分枝杆菌eis基因对巨噬细胞自噬的影响。方法将鼠巨噬细胞Raw264.7以自噬体荧光表达质粒GFP-LC3转染,将含eis基因的重组耻垢分枝杆菌MS—pmv261-eis与不含eis基因的耻垢分枝杆菌MS—pmv261分别感染宿主巨噬细胞,透射电镜下观察自噬小体形成情况,荧光显微镜下观察自噬荧光并计数,Westernblot检测如基因表达的蛋白及自噬蛋白LC3-Ⅱ的表达水平。结果结核分枝杆菌eis基因可抑制感染宿主细胞自噬小体的形成,并显著抑制自噬荧光小点形成(P〈0.05),显著降低了自噬蛋白LC3-Ⅱ表达水平。结论结核分枝杆菌e曲基因对Raw264.7细胞自噬有抑制作用。  相似文献   
8.
The principal sulfatide of virulentMycobacterium tuberculosis, sulfolipid-I (SL-I), both directly stimulates neutrophil superoxide (O 2 ) release and, at substimulatory concentrations, primes these cells for markedly enhanced oxidative responsiveness to other stimuli. The present study was undertaken to clarify the priming mechanisms by comparing cellular events following priming doses of SL-I with those following priming with N-formyl-methionyl-leucyl-phenylalanine (FMLP). We compared the involvement of the calcium cation (Ca2+), as well as membrane protein kinase C (PKC) activity and the translocation of NADPH oxidase-cytosolic cofactor effected by priming levels of the two agonists. The investigation led to two important conclusions. First, we clearly demonstrate that priming by both SL-I and FMLP results from activation of cellular processes that are not involved in direct oxidative activation. For example, whereas direct induction of O 2 generation by FMLP and SL-I required increases in intracellular Ca2+, an increase in intracellular calcium concentration ([Ca2+]i) above basal levels was not required for priming. Second, we identified key differences in the cellular responses to priming doses of SL-I and FMLP. Whereas increased membrane PKC activity caused by priming doses of FMLP was only partially blocked by chelation of intracellular Ca2+, Ca2+ chelation completely inhibited the increase in membrane PKC activity caused by SL-I. NADPH oxidase-cytosolic factor translocation to plasma membranes was completely blocked by pertussis toxin when priming doses of SL-I were used. This guanine-nucleotide-binding protein inhibitor had no effect on FMLP-dependent translocation of the oxidase cofactors. The comparative approach introduced in this report provides a valuable and novel method to discern the complex interactions of various cellular processes that regulate the state of activation of stimulated cells.  相似文献   
9.
Since 1985 mycobacterial infection has been observed occasionally among snakes and bullfrogs housed in the Wisconsin exhibit at the Milwaukee Zoo. Prospective screening of animals was initiated in September 1990, after two cases occurred in March and June 1990. Overall, of 47 animals that were housed in the exhibit from 1981 through its closure in 1995, 15 (31.9%) were diagnosed with mycobacterial infection. That includes 10 cases (of 24 animals; 40% prevalence) that occurred during the final 5 years, when all animals were actively being screened for infection. Infection was documented by culture for seven animals, histology for four animals, and both histology and culture for four animals. Species determination of nine of the 10 isolates revealed Mycobacterium marinum. Genetic fingerprinting of the eight available isolates using pulsed field gel electrophoresis (PFGE) showed that six animals (five snakes and one bullfrog) were infected with the same strain of M. marinum (strain A) and two snakes were infected with a second strain (strain B). Deaths of animals infected with strain A spanned 1992–1995, and for strain B 1990–1992. It is postulated that possible routes of transmission were inhalation of infected aerosols or ingestion of contaminated food, water, or fomites. These data suggest that in closed systems the presence of mycobacterial infection in one animal significantly increases the risk of infection for all animals. Moreover, individual pathogenic strains may persist for prolonged periods of time. Zoo Biol 21:233–241, 2002. © 2002 Wiley‐Liss, Inc.  相似文献   
10.
利用rpoB基因芯片技术进行快速分枝杆菌菌种鉴定   总被引:2,自引:0,他引:2  
利用rpoB基因芯片技术快速进行分枝杆菌菌种鉴定。以分枝杆菌rpoB基因编码序列为靶基因, 用基因芯片技术检测21种分枝杆菌标准株;8种其它细菌标准株;126株临床分离株。分枝杆菌与其它细菌标准株经PCR扩增后, 分枝杆菌标准株均扩增出360 bp DNA片段, 在其它细菌中, 除甲型溶血性链球菌和假白喉棒状杆菌出现同样片段外, 其它细菌均未见扩增。21种寡核苷酸探针除海分枝杆菌与偶然分枝杆菌的探针有交叉杂交外, 其余均为特异性杂交。对126株临床分离株进行鉴定, 89株为结核分枝杆菌, 占70.6%(89/126), 非结核分枝杆菌(NTM)占9.2%(9/98)。应用rpoB基因芯片技术鉴定分枝杆菌菌种, 是一种快速、准确的方法, 具有较高的临床应用价值。  相似文献   
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