Phylogenetic comparison of the S3 gene of United States prototype strains of bluetongue virus with that of field isolates from California.

To better define the molecular epidemiology of bluetongue virus (BTV) infection, the genetic characteristics and phylogenetic relationships of the S3 genes of the five U.S. prototype strains of BTV, the commercially available serotype 10 modified live virus vaccine, and 18 field isolates of BTV serotypes 10, 11, 13, and 17 obtained in California during 1980, 1981, 1989, and 1990 were determined. With the exception of the S3 gene of the U.S. prototype strain of BTV serotype 2 (BTV 2), these viruses had an overall sequence homology of between 95 and 100%. Phylogenetic analyses segregated the prototype U.S. BTV 2 strain to a unique branch (100% bootstrap value), whereas the rest of the viruses clustered in two main monophyletic groups that were not correlated with their serotype, year of isolation, or geographical origin. The lack of consistent association between S3 gene sequence and virus serotype likely is a consequence of reassortment of BTV gene segments during natural mixed infections of vertebrate and invertebrate hosts. The prototype strain of BTV 13, which is considered an introduction to the U.S. like BTV 2, presents an S3 gene which is highly homologous to those of some isolates of BTV 10 and especially to that of the vaccine strain. This finding strongly suggests that the U.S. prototype strain of BTV 13 is a natural reassortant. The different topologies of the phylogenetic trees of the L2 and S3 genes of the various viruses indicate that these two genome segments evolve independently. We conclude that the S3 gene segment of populations of BTV in California is formed by different consensus sequences which cocirculate and which cannot be grouped by serotype.     

Expression of a full-length nonstructural protein NS1 of bluetongue virus serotype 17 in Escherichia coli.

The relative abundance of the nonstructural protein NS1 in bluetongue virus (BTV)-infected cells, the existence of NS1 in the BTV particles and the highly conserved NS1 gene among BTV serotypes indicate the diagnostic potential of using NS1 in detecting BTV infections. In this study a NS1 gene was expressed with the T7 RNA polymerase expression system to produce a full-length NS1 protein. Sheep anti-NS1 antibodies were raised with the E. coli-produced NS1 and used to show that the NS1 proteins of the five BTV serotypes in the Unites States were immunologically indistinguishable.     

Impact of vacuum cooling on Escherichia coli O157:H7 infiltration into lettuce tissue

Vacuum cooling is a common practice in the California leafy green industry. This study addressed the impact of vacuum cooling on the infiltration of Escherichia coli O157:H7 into lettuce as part of the risk assessment responding to the E. coli O157:H7 outbreaks associated with leafy green produce from California. Vacuum cooling significantly increased the infiltration of E. coli O157:H7 into the lettuce tissue (2.65E+06 CFU/g) compared to the nonvacuumed condition (1.98E+05 CFU/g). A stringent surface sterilization and quadruple washing could not eliminate the internalized bacteria from lettuce. It appeared that vacuuming forcibly changed the structure of lettuce tissue such as the stomata, suggesting a possible mechanism of E. coli O157:H7 internalization. Vacuuming also caused a lower reduction rate of E. coli O157:H7 in stored lettuce leaves than that for the nonvacuumed condition.     

Peptide binding identifies an ERalpha conformation that generates selective activity in multiple in vitro assays

Drugs such as tamoxifen, which act at the estrogen receptor (ER), have very different in vitro and in vivo effects from those of the native hormone. Previous research has established that different ligands induce distinct conformational changes in the ER, thus affecting the interactions of the receptor with cell-specific co-activating or co-repressing proteins (cofactors) and estrogen response elements (EREs), thus potentially driving differing biological effects. Affinity-selected peptides have been used to probe the conformational changes that occur within the ER upon binding various ligands. In this study, the authors characterize the ability of several peptides to be recruited to liganded ER under cellular conditions. Approximating ER conformation via recruitment of this peptide to the ER is concluded to be a better predictor of the agonist nature of an ER ligand under these different cellular contexts than is a canonical cotransfection transactivation assay.     

‘Follow the Water’: Hydrogeochemical Constraints on Microbial Investigations 2.4 km Below Surface at the Kidd Creek Deep Fluid and Deep Life Observatory

Abstract

Microbiological and geochemical data are presented to characterize the hydrogeochemistry and to investigate extant microbial life in fracture waters 2.4?km below surface, at the Kidd Creek Observatory in Canada. Previous studies identified the world’s oldest groundwaters with mean residence times on the order of millions to billions of years trapped in fractures in Precambrian host rock here. In this study, major ion chemistry, δ¹⁸O and δ²H isotopic signatures and dissolved gases in the fracture waters are shown to be distinct from potential contamination end-members, demonstrating the fracture waters are not impacted by waters used in mining operations. A previous work on sulfur isotope signatures suggested a longstanding indigenous population of sulfate-reducing bacteria in these highly reducing fluids and sufficient sulfate to support microbial activity. Here, we report the first evidence for extant visible and cultivable microbial life at this location. Anaerobic metabolisms were investigated using the Most Probable Number (MPN) method. The fracture fluids contained extant cells at low biomass density (～10³–10⁴ cells/mL) and showed a strong response from autotrophic sulfate-reducers and alkane-oxidizing sulfate reducers. These lines of evidence provide the interpretational framework (chemical, hydrogeologic, and microbiologic) essential to the on-going genomic and metagenomic investigations at the Kidd Creek Observatory – the world’s most longstanding location for investigation of subsurface fluids and deep life at such profound depth.     

桑螵蛸挥发油的提取及对MRSA体外抑菌效应的初步研究

目的研究桑螵蛸挥发油提取物对耐甲氧西林金黄色葡萄球菌(MRSA)的体外抑菌效应。方法采用索氏提取法提取桑螵蛸中挥发油成分,在鉴定耐药基因基础上,再采用纸片扩散法(K-B法)、试管稀释法(MIC法)、吸光度值测定法来观察桑螵蛸挥发油提取物对MRSA的体外抑菌效应。结果桑螵蛸挥发油提取物对MRSA的生长表现出抑制效应。结论桑螵蛸挥发油提取物对MRSA具有一定的抑菌效应,为下一步研究其详细有效成分及开发提供数据和理论依据。     

呼伦贝尔草原辉河湿地不同淹水状态的土壤碳氮磷特征比较

碳(C)、氮(N)、磷(P)生态化学计量比是生态系统过程及其功能的重要特征。为了比较不同淹水状态下草原区河岸湿地土壤生态化学计量特征差异,探讨用土壤生态化学计量特征来表征草原河岸湿地的退化状态,以呼伦贝尔草原辉河河岸湿地为研究对象,分别选择完全退化和未退化河岸湿地,从中选取常年淹水区域(W)、年际干湿交替区域(WD)以及河岸带边缘多年未曾淹没的干燥区域(D)为观测样地,对比研究退化与未退化草原河岸湿地土壤C、N、P化学计量比在不同淹水状态下的表征。结果表明:(1)随着淹水状态的改变,不论是退化河岸湿地还是未退化河岸湿地,不同淹水状态下的土壤C/N、C/P、N/P均差异显著(P0.05);(2)退化河岸湿地的C/N、C/P和N/P的平均值分别为9.05、273.08和28.25,与未退化河岸湿地相比(7.85、95.48、11.41)值显著升高;(3)相关性分析结果显示,土壤总磷与有机碳、全氮显著正相关,与C/N、C/P、N/P负相关,这可能是限制退化河岸湿地土壤养分元素循环的主要因素。研究结果表明:呼伦贝尔草原辉河河岸湿地不同淹水状态的土壤碳、氮、磷特征差异明显(P0.05),尤其是土壤C/P和N/P。与未退化河岸湿地相比,退化河岸湿地的C/N、C/P、N/P均明显高于未退化河岸湿地(P0.05),因此可以将土壤碳、氮、磷化学计量比特征作为草原区河岸湿地退化的评价指标。