Production of α-ketoisocaproate via free-whole-cell biotransformation by Rhodococcus opacus DSM 43250 with L-leucine as the substrate

This work aims to produce α-ketoisocaproate (KIC) from l-leucine via the free-whole-cell biotransformation of Rhodococcus opacus DSM 43250. The effects of temperature, pH, substrate concentration, cell concentration, and rotating speed on KIC production were examined. Furthermore, the biotransformation conditions were optimized with response surface methodology (RSM). The optimal biotransformation conditions were as follows: temperature 43.7 °C, pH 8.4, l-leucine concentration 5.1 g/L, cell concentration 30.4 g/L, and rotating speed 170 rpm. The maximal KIC production predicted by RSM model reached 1275 mg/L at the optimal conditions, and in the validated experiments, the maximal KIC production reached 1264 mg/L, which was very close to the predicted KIC production. The current work provides an alternative for the production of KIC and the results obtained here are useful for the biotransformatic KIC production on a large scale.     

杭州湾滨海滩涂盐基阳离子对植物分布及多样性的影响

滨海滩涂由于其高含盐量显著影响了植物群落分布及生物多样性。目前有关滩涂含盐量与生物多样性的关系研究较多,但不同区域盐基离子组成不同,且对植物的影响也存在较大差异,以杭州湾南岸不同年代形成的滩涂为研究对象,系统监测了50个样方土壤交换性盐基阳离子的组成、分布和植物组成及多样性特征等,采用去趋势典范对应分析(DCCA)、线性回归和多元逐步回归分析了4种盐基阳离子对物种数量、分布和多样性的影响。结果表明,杭州湾南岸滩涂4种主要盐基阳离子含量(g/kg)大小顺序为Ca²⁺>Na⁺>Mg²⁺>K⁺,其中Ca²⁺占到总含量的61.97%;经DCCA分析发现4种盐基阳离子对植物群落的分布均有显著影响,但以Ca²⁺的影响程度最大;随着盐基离子含量的逐渐降低,物种数量逐渐增加,多样性指数逐渐增加,同时也发现Ca²⁺对两种多样性指数影响最大。     

Improvement of laccase production and its properties by low-energy ion implantation

Low-energy ion implantation was employed to breed laccase producing strain Paecilomyces sp. WSH-L07 and a mutant S152 that exhibited an activity of more than three times over the wild strain was obtained. The optimum substrate of both the wild and mutant laccases was 2,2′-azino-bis(3-ethylbenzothiazoline-6-sulfonate), and followed by guaiacol with optimal pH at 3.4 and 5.0, respectively, while the mutant laccase exhibited a broader active pH range. The mutant laccase had a higher optimal catalytic temperature (60–65 °C) than the wild one (55 °C), and the wild laccase deactivated rapidly when temperature increased above 55 °C. Furthermore, the mutant laccase was more stable under neutral and alkaline conditions. A thermostability experiment revealed that the mutant laccase was superior to the wild laccase. Both laccases were stable in the presence of metal ions, mildly inhibited by SDS (0.5 mM), EDTA (1 mM) and 1,4-dithiothreitol (0.5 mM), and almost completely inhibited by 0.1 mM NaN₃.     

气候变化情景下大沙鼠潜在地理分布

大沙鼠（Rhombomys opimus）是中亚地区典型的荒漠啮齿动物,其采食和掘洞行为造成了荒漠林和荒漠草原退化加剧,生态环境恶化。基于大沙鼠分布数据、气候、土壤和地形因子数据,采用MaxEnt模型预测大沙鼠在当前气候和温室气体低、中、高3种浓度排放情景下2050年和2070年的潜在适生区,分析亚洲大陆未来气候条件下大沙鼠适生面积和分布格局的变化趋势,探讨影响大沙鼠分布的主要环境因子。结果表明：模型AUC（Area Under Curve）值达到0.9以上,预测的准确性达到"极好"。经刀切法分析（Jackknife）表明,影响大沙鼠在适生区分布最主要的环境变量为温度季节性变化的标准差、土壤基本饱和度、最干季度降水量、最暖季度降水量和土壤可交换钠盐。Rcp2.6、Rcp4.5和Rcp8.5三种气候场景下2050年高适生区面积较当前分别增长15.78%、15.10%和13.44%;Rcp2.6、Rcp4.5和Rcp8.5三种气候场景下2070年高适生区面积较当前增长8.32%、13.18%和18.18%。中国大沙鼠适生区范围内,新疆所分布的大沙鼠适生区分布范围变化较大,3种情景模式下大沙鼠的适生区位置向新疆北部扩张;甘肃适生区位置向西北部扩张;内蒙西北部和阿拉善地区大沙鼠的适生区位置向四周扩张。研究揭示了未来气候下大沙鼠高适生区范围和空间变化,并得到影响其分布的主要环境变量,对其防控具有重要意义。     

hSHIP induces S-phase arrest and growth inhibition in cervical cancer HeLa cells

hSHIP,a human SH2-containing inositol-5-phosphatase,acts as a negative regulator of proliferation and survival in hematopoietic cells.Therefore,hSHIP may play a crucial role in suppression of cervical cancer HeLa cells.In this study,pcDNA3.1-hSHIP-GFP plasmid was constructed and transfected into HeLa cells with Lipofectamine2000,stably transfected HeLa cells were established and their responses were investigated by Flow cytometry,MTT,tumorigenicity in nude mice,RT-PCR and ELISA assays.The results showed tha...     

Hela细胞P16~(ink4)cDNA的克隆及序列分析

P16~(ink4)是CDK抑制蛋白,参与调控细胞G1期至S期的转换。目前发现P16`(ink4)基因损伤与多种肿瘤的发生、发展有关,可能是功能上非常重要的抑癌基因。为了研究该基因的功能,以及突变对该基因功能的影响,本文应用RT-PCR方法,从Hela细胞中克隆了P16~(ink4)cDNA。扩增得到556bp片段(包括引物两端酶切位点的16bp)克隆于M13载体,测定了其DNA序列。该序列包括了P16~(ink4)cDNA编码区全部471bp,以及3’端69bp序列。表明P16~(ink4)cDNA已成功地得到克隆。     

Biosynthesis of flower-shaped Au nanoclusters with EGCG and their application for drug delivery

Background

In the last decade, the biosynthesis of metal nanoparticles using organisms have received more and more considerations. However, the complex composition of organisms adds up to a great barrier for the characterization of biomolecules involved in the synthesis process and their biological mechanisms.

Results

In this research, we biosynthesized a kind of flower-shaped Au nanoclusters (Au NCs) using one definite component—epigallocatechin gallate (EGCG), which was the main biomolecules of green tea polyphenols. Possessing good stability for 6 weeks and a size of 50 nm, the Au NCs might be a successful candidate for drug delivery. Hence, both methotrexate (MTX) and doxorubicin (DOX) were conjugated to the Au NCs through a bridge of cysteine (Cys). The introduction of MTX provided good targeting property for the Au NCs, and the conjugation of DOX provided good synergistic effect. Then, a novel kind of dual-drug loaded, tumor-targeted and highly efficient drug delivery system (Au-Cys-MTX/DOX NCs) for combination therapy was successfully prepared. The TEM of HeLa cells incubated with Au-Cys-MTX/DOX NCs indicated that the Au-Cys-MTX/DOX NCs could indeed enter and kill cancer cells. The Au-Cys-MTX/DOX NCs also possessed good targeting effect to the FA-receptors-overpressed cancer cells both in vitro and in vivo. Importantly, the Au-Cys-MTX/DOX NCs resulted in an excellent anticancer activity in vivo with negligible side effects.

Conclusions

These results suggest that the biosynthesized Au-Cys-MTX/DOX NCs could be a potential carrier with highly efficient anticancer properties for tumor-targeted drug delivery.

     

Using automated electronic medical record data extraction to model ALS survival and progression

Background

To assess the feasibility of using automated capture of Electronic Medical Record (EMR) data to build predictive models for amyotrophic lateral sclerosis (ALS) outcomes.

Methods

We used an Informatics for Integrating Biology and the Bedside search discovery tool to identify and extract data from 354 ALS patients from the University of Kansas Medical Center EMR. The completeness and integrity of the data extraction were verified by manual chart review. A linear mixed model was used to model disease progression. Cox proportional hazards models were used to investigate the effects of BMI, gender, and age on survival.

Results

Data extracted from the EMR was sufficient to create simple models of disease progression and survival. Several key variables of interest were unavailable without including a manual chart review. The average ALS Functional Rating Scale – Revised (ALSFRS-R) baseline score at first clinical visit was 34.08, and average decline was ??0.64 per month. Median survival was 27?months after first visit. Higher baseline ALSFRS-R score and BMI were associated with improved survival, higher baseline age was associated with decreased survival.

Conclusions

This study serves to show that EMR-captured data can be extracted and used to track outcomes in an ALS clinic setting, potentially important for post-marketing research of new drugs, or as historical controls for future studies. However, as automated EMR-based data extraction becomes more widely used there will be a need to standardize ALS data elements and clinical forms for data capture so data can be pooled across academic centers.

     

Surfactant protein D attenuates nitric oxide-stimulated apoptosis in rat chondrocyte by suppressing p38 MAPK signaling

Innate immune molecule surfactant protein D (SP-D), a member of the C-type lectin protein family, plays an indispensable role in host defense and the regulation of inflammation in the lung and other tissues. Osteoarthritis (OA) is a degenerative disease of cartilage, with inflammation that causes pathologic changes and tissue damage. However, it is unknown whether there exist SP-D expression and its potential role in the pathogenesis of OA. In this study, we examined SP-D expression and explored its biological function in a sodium nitroprusside (SNP)-stimulated rat chondrocytes and surgically-induced rat OA model. We found SP-D expression in both human and rat articular chondrocytes, with higher level in normal chondrocytes compared to in OA chondrocytes. Furthermore, In vivo study demonstrated that recombinant human SP-D (rhSP-D) ameliorated cartilage degeneration in surgically-induced rat OA model. In vitro cell culture study showed that rhSP-D markedly inhibited the expression of caspase-3 as an apoptosis biomarker, and decreased phosphorylation of p38 mitogen-activated protein kinase (MAPK), which resulted in maintaining normal nuclear morphology and increasing mitochondrial membrane potential in SNP-stimulated rat chondrocytes. Collectively, these findings indicate that SP-D expresses in articular chondrocytes and suppresses SNP-stimulated chondrocyte apoptosis and ameliorates cartilage degeneration via suppressing p38 MAPK activity.