RFLP analysis of SLA haplotypes in Swiss large white and American Hampshire pigs using SLA class I and class II probes

Summary

Restriction fragment length polymorphism (RFLP) analyses of swine leukocyte antigen (SLA) class I and class II genes from Swiss Large White and American Hampshire families were performed using porcine DNA probes. Class I and class II RFLPs associated with the serologically‐defined haplotypes SLA H1, H8, H16 and H24 and with serotypes SLA 15, 16; SLA 14; and SLA 6, SB 19, were identified. Seven allelic class I RFLP patterns were observed. For genes in the SLA class II region, six allelic RFLP patterns of DQA and DQB; five allelic RFLP patterns of DRA; and seven allelic RFLP patterns of DRB were observed. The serologically‐defined H8 haplotype was subtyped based on differences in class II RFLPs.     

粪便微生物宏基因组来源GH1 β-葡萄糖苷酶的重组表达及酶学性质

[背景] β-葡萄糖苷酶是一类重要的纤维素分解酶类。目前较多可培养微生物来源的β-葡萄糖苷酶（Bgl）存在热稳定性和酸碱稳定性差及作用范围窄的问题。[目的] 从滇金丝猴粪便微生物宏基因组中挖掘新型β-葡萄糖苷基因,异源表达并研究其酶学性质,为食品等领域提供新型酶资源。[方法] 从粪便微生物宏基因组出发,扩增和异源表达β-葡萄糖苷酶基因BglRBS_26和BglRBS_9,并进行酶学性质研究。[结果] 获得GH1家族重组β-葡萄糖苷酶BglRBS_26和BglRBS_9,分子量分别为60 kD和50 kD。BglRBS_26的最适作用条件为pH 6.0、45 ℃;BglRBS_9的最适作用条件为pH 5.0、40 ℃。BglRBS_26和BglRBS_9的K_m分别为0.681 6±0.164 2 μmol/L和3.317 0±0.871 4 μmol/L。BglRBS_26具有较好的酸碱耐受性,在pH 5.0–6.0下处理1 h,剩余酶活大于110%;pH 7.0-8.0范围内,剩余酶活仍然保持在100%以上。此外,蔗糖对BglRBS_26和BglRBS_9有不同程度的激活,当反应体系中加入20%（质量体积分数）蔗糖,BglRBS_26酶活力可提高至140%;10%（质量体积分数）蔗糖可将BglRBS_9酶活力提高至180%。此外,BglRBS_26具有较好的NaCl耐受性和稳定性,其在37 ℃、2.5 mol/L的NaCl下处理1 h后保留80%活性。[结论] 从滇金丝猴粪便微生物宏基因组中获得2个新型β-葡萄糖苷酶基因BglRBS_26和BglRBS_9,并成功在大肠杆菌BL21（DE3）中表达。BglRBS_26和BglRBS_9具有较好的pH稳定性和蔗糖耐受性,在食品、发酵等行业具有潜在应用价值。     

Design and evaluation of novel 8-oxo-pyridopyrimidine Jak1/2 inhibitors

A highly ligand efficient, novel 8-oxo-pyridopyrimidine containing inhibitor of Jak1 and Jak2 isoforms with a pyridone moiety as the hinge-binding motif was discovered. Structure-based design strategies were applied to significantly improve enzyme potency and the polarity of the molecule was adjusted to gain cellular activity. The crystal structures of two representative inhibitors bound to Jak1 were obtained to enable SAR exploration.     

Detection of matrix metalloproteinase (MMP)-2 and MMP-9 in canine seminal plasma

Matrix metalloproteinases (MMPs) are a family of proteolytic enzymes that play a central role in degradation of protein components of the extracellular matrix and basement membrane. Previous studies have shown that MMP-2 and MMP-9 are present in human seminal plasma, but there is little information available on the presence of MMPs in canine seminal plasma. This study aims to investigate the presence of MMPs in canine seminal plasma and their clinical manifestation at the level of various semen parameters in canine species. Latent and active forms of MMP-2 and MMP-9 were evaluated using gelatin zymography and their association with semen parameters was examined. Results demonstrate that both latent and active forms of MMP-2 and MMP-9 are present in canine seminal plasma and the latent forms are predominant. The latent and active MMP-9 activities were elevated in the semen with unsatisfactory quality traits and proMMP-2 was inversely correlated with semen quality whereas, MMP-2 was positively correlated with semen quality traits. These findings suggest that proMMP-9 and MMP-9 activation contributes to the variation in semen, while the activation of MMP-2 improves the sperm functionality.     

Identification of cancer genomic markers via integrative sparse boosting

In high-throughput cancer genomic studies, markers identified from the analysis of single data sets often suffer a lack of reproducibility because of the small sample sizes. An ideal solution is to conduct large-scale prospective studies, which are extremely expensive and time consuming. A cost-effective remedy is to pool data from multiple comparable studies and conduct integrative analysis. Integrative analysis of multiple data sets is challenging because of the high dimensionality of genomic measurements and heterogeneity among studies. In this article, we propose a sparse boosting approach for marker identification in integrative analysis of multiple heterogeneous cancer diagnosis studies with gene expression measurements. The proposed approach can effectively accommodate the heterogeneity among multiple studies and identify markers with consistent effects across studies. Simulation shows that the proposed approach has satisfactory identification results and outperforms alternatives including an intensity approach and meta-analysis. The proposed approach is used to identify markers of pancreatic cancer and liver cancer.     

Design, synthesis, and anticancer activity of phosphonic acid diphosphate derivative of adenine-containing butenolide and its water-soluble derivatives of paclitaxel with high antitumor activity

Synthesis of adenine derivative of triphosphono-gamma-(Z)-ethylidene-2,3-dimethoxybutenolide 4 was accomplished by treatment of phosphonate 3 with 5-phosphoribosyl 1-pyrophosphate in the presence of 5-phosphoribosyl 1-pyrophosphate synthetase. It was found that triphosphonate 4 functions as an irreversible stoichiometric inactivator of the Escherichia coli ribonucleoside diphosphate reductase (RDPR). Triphosphonate 4 exhibited potent inhibitory activity against murine leukemias (L1210 and P388), breast carcinoma (MCF7), and human T-lymphoblasts (Molt4/C8 and CEM/0) cell lines. Paclitaxel ester derivatives of adenine-containing triphosphono-gamma-(Z)-ethylidene-2,3-dimethoxybutenolide 8-10 were also synthesized. Like triphosphonate 4, compound 8 exhibited inhibitory property toward RDPR. It also induced microtubule assembly similar to paclitaxel (5). The structure of the chlorodiester linker in 8 was found to account for this dual property. After treatment of MCF7 cells with compounds 4, 5, and 8, fluorescence microscope examination demonstrated the presence of nucleus shrinkage or segmentation. Bifunctional prodrug 8 exhibited higher lipophilicity than 4 and higher water-solubility than 5. Pro-dual-drug 8 exhibited more pronounced anticancer activity relative to that of the triphosphonate 4 and paclitaxel (5). In contrast, compound 9, resulting from the linkage of triphosphonate 4 and paclitaxel (5) through a diester unit, was only found to function as a highly water-soluble prodrug for paclitaxel (5). It induced microtubule assembly in vitro, but did not show inhibitory property toward RDPR. On the other hand, compound 10, an aggregate of triphosphonate 4 and paclitaxel (5), neither functioned as an inhibitor of RDPR nor exhibited microtubule assembly stimulating activity in vitro.     

Evidence of Association of Salmonellae with Tomato Plants Grown Hydroponically in Inoculated Nutrient Solution

The possibility of uptake of salmonellae by roots of hydroponically grown tomato plants was investigated. Within 1 day of exposure of plant roots to Hoagland nutrient solution containing 4.46 to 4.65 log₁₀ CFU of salmonellae/ml, the sizes of the pathogen populations were 3.01 CFU/g of hypocotyls and cotyledons and 3.40 log₁₀ CFU/g of stems for plants with intact root systems (control) and 2.55 log₁₀ CFU/g of hypocotyls and cotyledons for plants from which portions of the roots had been removed. A population of ≥3.38 log₁₀ CFU/g of hypocotyls-cotyledons, stems, and leaves of plants grown for 9 days was detected regardless of the root condition. Additional studies need to be done to unequivocally demonstrate that salmonellae can exist as endophytes in tomato plants grown under conditions that simulate commonly used agronomic practices.     

雌激素和孕激素对雌性小鼠甲状腺C细胞的影响

探讨降钙素在老年女性骨质疏松症发病中的作用 ,观察了雌、孕激素对雌性小鼠甲状腺 C细胞的影响。去卵巢小鼠 (OVX)分别肌肉注射苯甲酸雌二醇 (FB)、己烯孕酮 (HPC)和苯甲酸雌二醇加乙烯孕酮 (EB+HPC)两个月。用药剂量根据小鼠口龄和体重的不同而不同。采用免疫组织化学方法显示降钙素 (CT)阳性细胞并对其进行细胞形态计量学分析。结果显示 :(1)去卵巢小鼠与正常对照组相比 ,甲状腺 C细胞数目剧增 ,给予 EB、HPC、EB+HPC治疗的各组小鼠甲状腺C细胞数目与正常对照组水平相近。 (2 )各组小鼠甲状腺 C细胞的大小无明显改变 ,平均直径间无显著性差异 (P>0 .0 5 )。雌、孕激素缺乏可引起雌性小鼠甲状腺 C细胞增生。 C细胞增生也可能是雌激素缺乏引起的骨质疏松的后果