Interplay between Selenium,Selenoproteins and HIV-1 Replication in Human CD4 T-Lymphocytes

The infection of CD4 T-lymphocytes with human immunodeficiency virus (HIV), the etiological agent of acquired immunodeficiency syndrome (AIDS), disrupts cellular homeostasis, increases oxidative stress and interferes with micronutrient metabolism. Viral replication simultaneously increases the demand for micronutrients and causes their loss, as for selenium (Se). In HIV-infected patients, selenium deficiency was associated with a lower CD4 T-cell count and a shorter life expectancy. Selenium has an important role in antioxidant defense, redox signaling and redox homeostasis, and most of these biological activities are mediated by its incorporation in an essential family of redox enzymes, namely the selenoproteins. Here, we have investigated how selenium and selenoproteins interplay with HIV infection in different cellular models of human CD4 T lymphocytes derived from established cell lines (Jurkat and SupT1) and isolated primary CD4 T cells. First, we characterized the expression of the selenoproteome in various human T-cell models and found it tightly regulated by the selenium level of the culture media, which was in agreement with reports from non-immune cells. Then, we showed that selenium had no significant effect on HIV-1 protein production nor on infectivity, but slightly reduced the percentage of infected cells in a Jurkat cell line and isolated primary CD4 T cells. Finally, in response to HIV-1 infection, the selenoproteome was slightly altered.     

牛背最长肌高铁肌红蛋白还原酶的分离及其酶学性质

以冷鲜牛背最长肌为原料,采用硫酸铵盐析、超滤法对原料肉中的高铁肌红蛋白还原酶进行粗分离,并研究温度、pH、金属离子对高铁肌红蛋白还原酶活性的影响.结果表明:在硫酸铵饱和度65％,超滤(＞50 kDa)条件下,牛背最长肌高铁肌红蛋白还原酶可得到分离与纯化.试验范围内,牛背最长肌高铁肌红蛋白还原酶的分子量为29.0～66....     

酵母发酵法去除木糖母液中葡萄糖的研究

以木糖母液为原料,采用酵母发酵法去除母液中的葡萄糖。通过单因素分析,对影响去除效果的主要因素进行了研究。利用正交试验筛选出最佳工艺参数。结果表明:母液浓度为12%,调pH值4.5,酵母添加量2%,30℃下发酵6h,葡萄糖的去除率可达96%以上。     

A mutation in the COX5 gene of the yeast Scheffersomyces stipitis alters utilization of amino acids as carbon source, ethanol formation and activity of cyanide insensitive respiration

Scheffersomyces stipitis PJH was mutagenized by random integrative mutagenesis and the integrants were screened for lacking the ability to grow with glutamate as sole carbon source. One of the two isolated mutants was damaged in the COX5 gene, which encodes a subunit of the cytochrome c oxidase. BLAST searches in the genome of Sc. stipitis revealed that only one singular COX5 gene exists in Sc. stipitis, in contrast to Saccharomyces cerevisiae, where two homologous genes are present. Mutant cells had lost the ability to grow with the amino acids glutamate, proline or aspartate and other non-fermentable carbon sources, such as acetic acid and ethanol, as sole carbon sources. Biomass formation of the mutant cells in medium containing glucose or xylose as carbon source was lower compared with the wild-type cells. However, yields and specific ethanol formation of the mutant were much higher, especially under conditions of higher aeration. The mutant cells lacked both cytochrome c oxidase activity and cyanide-sensitive respiration, whereas ADH and PDC activities were distinctly enhanced. SHAM-sensitive respiration was obviously essential for the fermentative metabolism, because SHAM completely abolished growth of the mutant cells with both glucose or xylose as carbon source.     

改性大豆分离蛋白在黑胡椒油树脂复凝聚微胶囊制备中的应用

将大豆分离蛋白(soybean protein isolate,SPI)与木糖按不同质量比混合,于90℃下反应6 h,得到美拉德反应改性的SPI,再以改性SPI和壳聚糖为复合壁材,通过复凝聚法制备黑胡椒油树脂微胶囊,研究改性SPI对黑胡椒树脂微胶囊包埋效果、热稳定性等性质的影响。结果表明,当SPI/木糖质量比为2∶1时,黑胡椒油树脂微胶囊的包埋效率、产率及80℃下加热8 h后的保留率最高,分别为67.8%、72.07%和75.06%。热重分析表明,与天然SPI相比,改性SPI进一步提高了黑胡椒油树脂微胶囊的热稳定性,扫描电镜分析则表明改性SPI使微胶囊的微观结构更加致密;气相色谱-质谱分析表明利用改性SPI-壳聚糖制备的黑胡椒油树脂微胶囊对烯类挥发性成分的保持能力增加。本研究为拓展SPI的应用领域、提高黑胡椒油树脂微胶囊的稳定性提供参考。     

Germin-like protein Cit s 1 and profilin Cit s 2 are major allergens in orange (Citrus sinensis) fruits

Oranges are clinically relevant allergenic foods. To date, orange allergens have not been characterized in detail. The study is aimed at analyzing the sensitization profile in orange-sensitized subjects with and without clinical allergy, and to identify orange allergens. Fifty-six sensitized subjects with self-reported reactions to orange were grouped into reactors (anaphylaxis or multiple episodes of immediate reactions and/or positive challenge tests) and non-reactors (negative open food challenge tests). Allergens were characterized by IgE immunoblotting, N-terminal sequencing, IgE-inhibition assays, and mediator release assays were performed to determine the allergenic potency of orange profilin. Of 56 subjects, 23 were classified as orange allergic showing mainly an oral allergy syndrome. Of 23 subjects classified as orange allergic, 22 were sensitized to profilin, Cit s 2. In patients with mono-sensitization to profilin in vitro histamine releases up to 75% from basophils were induced using orange extract and purified plant profilins. Of the allergic patients 78% were sensitized to germin-like protein, Cit s 1. Both allergens showed retained IgE reactivity in heat-processed orange juice. Interestingly, subjects with and without clinical allergy showed a comparable sensitization profile. Profilin and germin-like proteins are major orange allergens. The potential clinical relevance of orange profilin was indicated by its strong capacity to release histamine from basophils. However, a predominant sensitization to both allergens in subjects without symptoms also indicates a high frequency of clinically insignificant sensitization.     

托盘和真空包装对冷却猪肉中MetMb还原活性的影响

在冷却肉的贮藏过程中,其肌红蛋白(Mb)被氧化为高铁肌红蛋白(MetMb),导致了冷却肉的褐变。研究了托盘包装和真空包装条件下,冷却肉自身的还原系统活性变化规律及其对肉色稳定性的影响。在2周的贮藏期内,真空与托盘包装冷却肉糜肌浆蛋白粗提液的还原活性持续下降;但真空包装组MetMb还原酶粗提液还原活力第1天到第14天活性差异不显著,托盘包装组前后差异显著。真空包装组在贮藏期间MetMb含量持续下降,但托盘包装组MetMb含量则持续上升,说明无氧条件有利于MetMb的还原。托盘包装组TBA在贮藏期间持续上生,但真空包装组TBA变化不明显。结果表明,托盘包装不利于冷却肉自身MetMb还原能力的维持,而真空包装的无氧环境,有利于MetMb的还原和肉色的保持。     

在酿酒酵母中共表达sXYLA和XKS1及木糖发酵的初步研究

为改善重组酵母发酵木糖生产乙醇的能力,将定点突变改造后的Thermus thermophilus木糖异构酶基因sXYLA克隆到酵母表达载体pYX212并用于转化酸酒酵母Saccharomyces cerevisiae YPH499进行表达研究。酶活检测表明,改造后的木糖异构酶活性是未改造的1.91倍。在此基础上将改造后具有良好特性的木糖异构酶基因sXYLA和来自酸酒酵母的木酮糖激酶基因XKS1耦联,构楚得到重组表达质粒pYX-sXYLA- XKS1,在酿酒酵母YPH499中实现组成型共表达。结果表明,在84 h时重组菌发酵液酶活达到最高,木糖异构酶为0.624 U/mg蛋白,木酮糖激酶为0.688 U/mg蛋白。以葡萄糖和木糖为混合碳源初步进行半通氧发酵,代谢产物分析表明酸酒酵母重组菌木糖的消耗为4.75 g/L,乙醇的产量为0.839 g/L,分别比出发菌提高20.9%和14.8%,为酿酒酵母利用木糖发酵乙醇奠定基础。