Isolation, Characterization and Phylogenetic Analysis of an Aerobic Bacterium Capable of Degrading Bensulfuronmethyl

Summary An aerobic bacterium named strain BH was isolated from soil samples based on its bensulfuronmethyl-degrading characteristics using continuous enrichment cultures. The cells of the strain were non-motile, gram-positive short rods. Colonies formed on agar medium were round, smooth, sticky, white-yellow in colour and of butyrous consistency. Analyses of nutritional utilization in Biolog microplates, conventional phenotypic characteristics and 16S rRNA gene sequencing were consistent with assigning strain BH to the genus Brevibacterium. Growth of the cells and their ability to degrade bensulfuronmethyl were simultaneously monitored under different liquid medium conditions during 7 days of incubation. They degraded bensulfuronmethyl from 100 to 70.6 mg l⁻¹ in mineral M9 medium and exhibited more effective degradation in the presence of yeast extract, completely removing an initial concentration of 100 mg l⁻¹ and at best 80% of an initial concentration of 200 mg l⁻¹. Further studies are required to determine the potential use of the isolate in the disposal of bensulfuronmethyl residues in agriculture and industry.     

植物抗盐胁迫研究进展

盐胁迫严重制约了农业生产,解析盐胁迫机理受到关注。随着抑制消减杂交和基因表达序列分析等大规模表达基因鉴定技术在抗盐研究中的应用,盐碱胁迫下功能基因的获得量迅速增加。综述了近年来在酵母、拟南芥、水稻等生物上利用基因芯片等方法进行抗盐机理研究取得的成果,并介绍了结合系统生物学的方法进行抗盐研究取得的进展,以及植物抗盐胁迫研究的发展前景。     

可溶性Jagged-1/Fc嵌合蛋白诱导淋巴结细胞向CD4+CD25+ T细胞分化

直接用可溶性Jagged-1/Fc嵌合蛋白(Jagged-1/Fc)在体外诱导小鼠淋巴结细胞向CD4 CD25 T细胞分化.通过荧光标记单克隆抗体染色结合流式细胞术,观察不同剂量Jagged-1/Fc在不同时间对淋巴结细胞向CD4 CD25 T细胞分化的影响,观察Jagged-1/Fc诱导T细胞内细胞因子的变化;藉ELISA法检测Jagged-1/Fc诱导分化的T细胞分泌TGF-β1、IL-4和IL-10的水平.结果显示,超过500.0μg/L剂量的Jagged-1/Fc使CD4 CD25 T细胞百分比明显增高,诱导时间需要4～6天,抗Jagged-1单抗能抵消Jagged-1/Fc的诱导作用,用DAPT阻断Notch信号通路的活化也能抑制Jagged-1/Fc的诱导作用,Jagged-1/Fc诱导分化的T细胞培养上清中IL-4和IL-10的水平明显增高,TGF-β1无明显变化,胞内IL-4,IL-10,IL-2和TNF-α的水平也呈增高趋势.上述结果表明,可溶性Jagged-1/Fc嵌合蛋白在体外可诱导小鼠淋巴结细胞向CD4 CD25 调节性T细胞分化.     

A profound decrease in maternal arginine uptake provokes endothelial nitration in the pregnant rat

While a specific role for nitric oxide (NO) in inducing the hemodynamic alterations of pregnancy is somewhat controversial, it is widely accepted that excess NO is generated during pregnancy. L-Arginine is the sole precursor for NO biosynthesis. Among several transporters that mediate L-arginine uptake, cationic amino acid transporter-1 (CAT-1) acts as the specific arginine transporter for endothelial NO synthase. The present study was designed to test the hypothesis that, during pregnancy, when arginine consumption by the fetus is significantly increased, compensatory changes in maternal arginine uptake affect the endothelium. Uptake of radiolabeled arginine (L-[3H]arginine) by freshly harvested maternal aortic rings from pregnant rats decreased by 65 and 30% in mid- and late pregnancy, respectively, compared with those obtained from virgin animals. This decrease was associated with a significant increase in endothelial protein nitration (the footprint of peroxynitrite generation), as shown by both Western blotting and immunohistochemistry utilizing anti-nitrotyrosine antibodies, reflecting endothelial damage. Northern blot analysis revealed that steady-state aortic CAT-1 mRNA levels did not change throughout pregnancy, whereas CAT-1 protein abundance was significantly increased, peaking at mid-pregnancy. Protein content of protein kinase C (PKC)-alpha, which was previously shown to decrease CAT-1 activity, increased significantly in the pregnant animals and was associated with a significant increase in CAT-1 phosphorylation. Intraperitoneal injection of alpha-tocopherol, a PKC-alpha inhibitor, prevented the decrease in arginine transport and attenuated protein nitration. In conclusion, aortic arginine uptake is reduced during pregnancy, through posttranslational modulation of CAT-1 protein, presumably via upregulation of PKC-alpha. The aforementioned findings are associated with an increase in protein nitration and, therefore, in selected individuals, may lead to the development of certain forms of endothelial dysfunction, like preeclampsia.     

Characterization of a bifunctional HPr kinase/phosphorylase from Leuconostoc mesenteroides SY1

The hprK gene encoding bifunctional HPrK/P (kinase/ phosphorylase) was cloned from L. mesenteroides SY1, a strain isolated from kimchi. hprK was transcribed as a monocistronic gene. His-tagged HPrH16A and HPrK/P were produced in E. coli BL21(DE3) using pET26b(+) and purified. HPrK/P phosphorylation assay with purified proteins showed that the kinase activity of HPrK/P increased at slightly acidic pHs. Divalent cations such as Mg2+ and Mn2+ and glycolytic intermediates such as fructose-1, 6-bisphosphate (FBP) and phosphoenolpyruvate (PEP) increased the kinase activity of HPrK/P, but inorganic phosphate strongly inhibited it. Kinetic studies for the kinase activity of HPrK/P showed that the apparent Km values were 0.18 and 14.57 microM for ATP and HPr, respectively. The Km value for the phosphorylase activity of HPrK/P was 14.16 microM for P-Ser-HPr (HPr phosphorylated at the serine residue).     

Purification of GFP fusion proteins with high purity and yield by monoclonal antibody-coupled affinity column chromatography

GFP has often been used as a marker of gene expression, protein localization in living and fixed tissues as well as for protein targeting in intact cells and organisms. Monitoring foreign protein expression via GFP fusion is also very appealing for bioprocess applications. Many cells, including bacterial, fungal, plant, insect and mammalian cells, can express recombinant GFP (rGFP) efficiently. Several methods and procedures have been developed to purify the rGFP or recombinant proteins fused with GFP tag. However, most current GFP purification methods are limited by poor yields and low purity. In the current study, we developed an improved purification method, utilizing a FMU-GFP.5 monoclonal antibody (mAb) to GFP together with a mAb-coupled affinity chromatography column. The method resulted in a sample that was highly pure (more than 97% homogeneity) and had a sample yield of about 90%. Moreover, the GFP epitope permitted the isolation of almost all the active recombinant target proteins fused with GFP, directly and easily, from the crude cellular sources. Our data suggests this method is more efficient than any currently available method for purification of GFP protein.     

Fungal Communities in Decaying Sapwood and Heartwood of a Conifer <Emphasis Type="Italic">Keteleeria evelyniana</Emphasis>

Fungal communities in decaying sapwood and heartwood of K. evelyniana were demonstrated through construction of four 18 S rRNA gene libraries. The 210 sequenced clones were clustered into 11 subgroups, belonging to Basidiomycota (71.9%) and to Ascomycota (22.4%) and unclassified (1 subgroup; 5.7%). The heartwood displayed higher species richness than the sapwood. Basidiomycota were dominant in either the heartwood or the sapwood. Phylogenetically diverse Homobasidiomycetes were detected in the heartwood, contrary to the sapwood, where Heterobasidiomycetes were detected. Clones close to Spongipellis unicolor dominated in the heartwood (21 of 99 clones), while those close to Hydnochaete olivacea dominated in the sapwood (41 of 111 clones). The common species between the two parts were those related to S. unicolor, Calocera cornea, Debaryomyces hansenii, Davidiella tassiana, and Nomuraea rileyi and those from Chaetothyriomycetes.     

PGK1 induction by a hydrogen peroxide treatment is suppressed by antioxidants in human colon carcinoma cells

Few protein biomarkers for oxidative stress have been reported. In this study, we attempted to identify the proteins selectively overexpressed in human colon tumor cells by treating with hydrogen peroxide as oxidative stress. A proteomic analysis followed by western blotting showed that phosphoglycerate kinase 1 (PGK1) was induced by hydrogen peroxide in a dose-dependent manner, while its expression was suppressed by a co-treatment with delphinidin, a known antioxidant. Furthermore, several antioxidants, including alpha-tocopherol, butylated hydroxytoluene (BHT), and Trolox, also inhibited the PGK1 induction caused by hydrogen peroxide. The data suggest that PGK1 might be a potential protein biomarker of intracellular oxidative status.     

Soil carbon pool and fertility under natural evergreen broadleaved forest and its artificial regeneration forests in southern Sichuan Province, China

The measurement of total soil organic matter (SOM) is not sensitive enough to detect short and medium term changes, and thus meaningful fractions of SOM should be measured. The carbon management index (CMI) was shown to be a useful technique for describing soil fertility. Soil samples were collected from natural evergreen broadleaved forest and its artificial regeneration forests of Sassafras tzumu, Cryptomeria fortunei and Metasequoia glyptostroboides in southern Sichuan Province, China, to determine soil carbon fractions, available nutrients, enzyme activity and CMI. Regression analysis was used to determine the relationship between soil carbon fractions, CMI and fertility. The results showed that the contents of soil organic carbon, water-soluble carbon, microbial biomass carbon, labile carbon, non-labile carbon, hydrolysis-N, available-P and available-K, the activity of invertase, phosphatase and catalase, and CMI were ranked with different seasons and followed the order: natural evergreen broadleaved forest > Sassafras tzumu plantation > Metasequoia glyptostroboides plantation > Cryptomeria fortunei plantation. The soil carbon fractions and CMI were significantly positively (P < 0.05) correlated with available nutrients and enzyme activity. The results indicate that soil carbon fractions and CMI could be used to evaluate the soil fertility for natural evergreen broadleaved forest and its artificial regeneration forests.