DNA末端硫修饰依赖的体外大片段DNA连接法

【背景】DNA组装技术是基因组合成中的一个关键技术。探索低成本、高效率的基因组合成技术一直是合成生物学的重要研究领域。在某些细菌如变铅青链霉菌中,DNA上有磷硫酰化修饰(简称硫修饰),而在另一些细菌如天蓝色链霉菌中存在一种含有硫修饰识别结构域(sulfur-binding domain, SBD)的识别蛋白,可以特异性识别DNA上的硫修饰,这启发了我们发展出一种新的DNA组装技术。【目的】探究在DNA末端硫修饰的连接中,T4 DNA连接酶与SBD相融合蛋白和单独的T4 DNA连接酶相比,是否有更高的连接效率。【方法】根据同源重组原理,设计硫修饰引物,扩增硫修饰的DNA片段。构建T4 DNA连接酶与SBD融合蛋白的3种表达载体T4-linker-SBD(Hga)、T4-linker-SBD(Spr)和T4-linker-SBD(Mmo),表达纯化以上3种融合蛋白。比较3组浓度梯度(2.4、0.24、0.024 mg/mL) T4 DNA连接酶与融合蛋白在2.5 kb和8.0 kb DNA片段连接上的差异。【结果】DNA末端硫修饰的2.5kb和8.0kb的两端片段均能扩增,而且3种融合蛋白...     

姜黄素通过抑制凋亡信号调节激酶1减弱PM2.5短期暴露致大鼠子宫损伤

细颗粒物(PM2.5)是空气动力学直径≤2.5 μm的颗粒物,能诱发多种疾病.已有大量的流行病学调查证实,PM2.5能够损伤生殖系统,但其致病机制不明确,相关的研究也非常有限.为研究PM2.5短期暴露对大鼠子宫的损伤,以及姜黄素(curcumin,CRC)对其保护作用,本研究将50只雌性SD大鼠随机分为生理盐水对照组、...     

长江平原区乡村景观的结构、管理及其对土壤氮磷的影响

通过实地研究主要位于宜兴市域的区域乡村景观代表性样方,评价并阐明了人口密集的长江平原区乡村景观的结构、管理与土壤全氮、全磷密度和储量的关系.景观绘图是基于1m分辨率的IKONOS影像并采用生态立地分类及绘图标准,通过直接解译和实地检验对均质景观缀块进行分类和绘图;依据区域权重分层取样方法,在生态立地缀块中随机设定取样点进行土壤或底泥取样;通过自助法对12个样方重取样,用区域多变量最优化方法计算样方的区域权重,并结合不确定性分析模型评价区域土地利用/覆被、土壤全氮和全磷储量.结果表明:在85.24×103km2的长江平原区乡村景观面积范围内,0～30cm土壤全氮、全磷储量分别为29.87Tg N和 19.79Tg P.最大的5种土地利用/覆被的类型为水田、水产养殖、非渗漏性建筑用地、旱地1年生作物和闲置水域,占区域总面积的82.9%;其土壤全氮、全磷储量分别占区域总量的82.6%和80.8%.其中平原稻田面积为38.93×103km2,占总面积的45.5%;其0～30cm土壤全氮、全磷储量分别高达15.26Tg N和9.13Tg P,分别占总储量的51%和45%.揭示了人口密集的乡村景观中土地利用/覆被方式对区域土壤全氮、全磷的影响模式.这种在小尺度下对土地管理调查,土壤全氮、全磷及其它生态特征研究方法的精确度明显优于传统的基于30～1000m分辨率遥感影像的土地覆被研究.     

不同居群不同部位头花蓼总黄酮含量研究

以原产贵州、四川的10个头花蓼收集系为材料,采用紫外分光光度法,研究了各头花蓼收集系植株不同部位总黄酮含量的变异,以便找出总黄酮含量较高的头花蓼收集系及其头花蓼植株上总黄酮含量较高的部位,为头花蓼药业提供指导。结果表明,不同部位头花蓼总黄酮含量差异很大,其含量大小依次为老茎>花>嫩叶>老叶>嫩茎。头花蓼老茎、花中总黄酮含量在不同居群间差异显著,其它部位差异不显著。四川都江堰居群老茎总黄酮含量最高,达2.761%,显著高于其它收集系。     

Long-term stability of parameters of antioxidant status in human serum

Abstract

The antioxidant status of serum or plasma can be determined using several commercially available assays. Here, four different assays, total antioxidant status (TAS), its second-generation assay (TAS2), biological antioxidant potential (BAP), and enzymatic assay using horseradish peroxidase (EAOC), were applied on human serum samples to test the temperature stability of antioxidants, upon storage of serum for 12 months. The two or three most commonly used temperatures for storage, that is, ? 20, ? 70 (or ? 80), and ? 196°C, were selected. The general conclusion is that all assays were stable at the temperatures tested. In addition, there were almost no statistically significant differences between the samples stored at different temperatures. Only the rank order of the EAOC assay was not very good in samples stored at ? 20°C. Also three components contributing to the total antioxidant capacity, uric acid, creatinine and bilirubin, showed no statistically significant differences between the temperatures. Therefore, storage at ? 20°C is sufficient to maintain a proper assay outcome of most of the total antioxidant assays, although storage at ? 70/80°C is to be preferred for longer storage times.     

The molecular size of glycans liberated by hydrazinolysis from semliki forest virus proteins

The glycans of well characterized, [6-³H]galactose-labelled glycopeptides, GC-4 from bovine IgG₁ as well as GP-V-2 and GP-V-5 from α₁-acid glycoprotein, were liberated by hydrazinolysis. Molecular weights close to the expected values were observed by gel filtration. Desialated glycans of Semliki Forest virus proteins were likewise liberated by hydrazinolysis and subjected to gel filtration. Metabolically labelled [1-³H]galactose-oligosaccharides of the mixed viral proteins revealed an apparent molecular weight of 1800. The bi-antennary glycan liberated from the reference glycopeptide GC-4 was of 1750 daltons. A mixture of [2-³H]mannose-labelled E₁-and E₂-proteins of the virus contained L-type glycans of 1800 daltons (formerly called A-type), and M-type glycans of 1200 daltons (formerly called B-type). A fraction of the E₃-glycans isolated by affinity chromatography on Concanavalin A-Sepharose showed an average molecular weight of 2150, a value intermediate between the three- and four-antennary glycans liberated from the reference glycopeptides GP-V-5 and GP-V-2. The rest of the E₃-glycans were of 1850 daltons, a value close to the bi-antennary GC-4 glycan. We suggest that the comparatively large size of the E₃-glycans and the exposed position of E₃-proteins on the viral surface may be interrelated.     

Boosting Polysulfide Redox Kinetics by Graphene‐Supported Ni Nanoparticles with Carbon Coating

Lithium–sulfur batteries have attracted extensive attention because of their high energy density. However, their application is still impeded by the inherent sluggish kinetics and solubility of intermediate products (i.e., polysulfides) of the sulfur cathode. Herein, graphene‐supported Ni nanoparticles with a carbon coating are fabricated by directly carbonizing a metal–organic framework/graphene oxide composite, which is then dispersed on a commercial glass fiber membrane to form a separator with electrocatalytic activity. In situ analysis and electrochemical investigation demonstrate that this modified separator can effectively suppress the shuttle effect and regulate the catalytic conversion of intercepted polysulfides, which is also confirmed by density functional theory calculations. It is found that Ni–C sites can chemically interact with polysulfides and stabilize the radical S₃^?? through Ni? S bonds to enable fast dynamic equilibrium with S₆^2?, while Ni nanoparticles reduce the oxidation barrier of Li₂S and accelerate ion/electron transport. As a result, the corresponding lithium–sulfur battery shows a high cycle stability (88% capacity retention over 100 cycles) even with a high sulfur mass loading of 8 mg cm^?2 and lean electrolyte (6.25 µ L mg^?1). Surprisingly, benefitting from the improved kinetics, the battery can work well at ?50 °C, which is rarely achieved by conventional Li–S batteries.     

New Lithium Salt Forms Interphases Suppressing Both Li Dendrite and Polysulfide Shuttling

Lithium–sulfur batteries (LSBs) are considered promising candidates for the next‐generation energy‐storage systems due to their high theoretical capacity and prevalent abundance of sulfur. Their reversible operation, however, encounters challenges from both the anode, where dendritic and dead Li‐metal form, and the cathode, where polysulfides dissolve and become parasitic shuttles. Both issues arise from the imperfection of interphases between electrolyte and electrode. Herein, a new lithium salt based on an imide anion with fluorination and unsaturation in its structure is reported, whose interphasial chemistries resolve these issues simultaneously. Lithium 1, 1, 2, 2, 3, 3‐hexafluoropropane‐1, 3‐disulfonimide (LiHFDF) forms highly fluorinated interphases at both anode and cathode surfaces, which effectively suppress formation of Li‐dendrites and dissolution/shuttling of polysulfides, and significantly improves the electrochemical reversibility of LSBs. In a broader context, this new Li salt offers a new perspective for diversified beyond Li‐ion chemistries that rely on a Li‐metal anode and active cathode materials.     

Protective effect of rosemary (Rosmarinus officinalis) against diethylnitrosamine-induced renal injury in rats

Abstract

Context: The kidney plays a central role in detoxification and excretion of toxic metabolites, and therefore, is susceptible to toxicity by xenobiotics.

Objective: To investigate the protective effect of Rosmarinus officinalis (rosemary) powder and its essential (volatile) oil against diethylnitrosamine (DEN)-induced renal injury in rats.

Materials and methods: Phenolic and flavonoid components were characterised in rosemary powder using HPLC-UV instrument while rosemary essential oil (E.O) was investigated via GC-MS technique. In rat model, rosemary was administrated orally (in diet) for two months. Lipid profile, antioxidant biomarkers, kidney functions and histopathological examinations were assessed.

Results: Hesperidin (4878.88?ppm) and ellagic acid (403.57?ppm) are among the major phenolic and flavonoid constituents in rosemary powder. Camphor (18.36%) and α-pinene (12.74%) represent the main E.O active ingredients. Rats treated with rosemary E.O showed a significant elevation in serum HDL (28.28%) accompanied by a decrease in LDL (115.47%). A significant decrease in serum creatinine and urea was also reported (69.72 and 109.89%, respectively). Moreover, serum glutathione peroxidise (GSH-Px) activity has been significantly increased. Kidney histopathological examinations confirmed the protective effect against DEN-induced abnormalities.

Conclusion: Rosemary (powder/E.O) was able to reduce or even prevent the severity of diethylnitrosamine-induced renal dysfunction.     

Length-weight relationship of 74 fish species caught in the continental coast of Ecuador

This work analysis the length-weight relationships for fish species captured by artisanal fishery along the continental coast of Ecuador. Fishermen used the following fishing gear: beach seine nets (5.1–31.8 mm), shrimp trawl nets (21.8–31.8 mm) and bottom gill nets (54.1–203.2 mm). Sampling took place during the years 2012 and 2014. Frequency of sampling was monthly, and it was completed systematically from Southern toward Northern Provinces. Length-weight relationship (LWR) was realized by the power model (y = a × x^b), for species with a bivariate dataset >70. A total of 59 commercial and 15 non-commercial species were registered. For 74 species, our analysis including a higher total length not reported in LWR estimates previously. The determination coefficient (r²) value was always above 0.95.