Components and proteolytic processing sites of arylsulfatase B from human placenta.

Previous studies have shown that mature arylsulfatase B purified from human sources is composed of two non-identical chains with apparent molecular masses of 43 kDa and 8 kDa. Arylsulfatase B purified from human placenta in the present study, however, included another 7 kDa component that could be detected only by carbohydrate staining on reducing SDS-PAGE employing the Tris-Tricine system. The 43 kDa and 7 kDa components contained a carbohydrate moiety, but the 8 kDa one did not, as demonstrated by periodic acid-Schiff staining, Con-A lectin blotting, endo-glycosidase treatment and in vitro phosphorylation by UDP-N-acetylglucosamine: lysosomal enzyme N-acetylglucosamine 1-phosphotransferase. The purified arylsulfatase B migrated as a single polypeptide of 58 kDa on non-reducing SDS-PAGE, indicating that the three chains are linked by disulfide bonds. In order to determine the origin of the components, N-terminal sequencing of the isolated polypeptides was performed. As a result, the 43, 7 and 8 kDa components were found to commence with Ala-41, Ala-424 and Asp-466, respectively. These results suggest that after removal of the signal peptide, human arylsulfatase B undergoes proteolytic processing on at least two sites during maturation.     

Entrapment of microbial cells and organelles with hydrophilic urethane prepolymers

Summary Microbial cells and cellular organelles were immobilized by mixing aqueous suspensions of the biocatalysts with water-miscible urethane prepolymers. Thus immobilized preparations of acetone-dried cells of Arthrobacter simplex and thawed cells of Nocardia rhodocrous showed appreciable {ie351-1} activities in the transformation of hydrocortisone into prednisolone and 4-androstene-3,17-dione to androst-1,4-diene-3,17-dione, respectively. The activities of catalase and alcohol oxidase were observed in the immobilized peroxisomes (microbodies) of a methanol-grown yeast Kloeckera sp. No. 2201. Yeast mitochondria entrapped with the prepolymer showed adenylate kinase activity. These results indicate the usefulness of the urethane prepolymers as convenient materials for entrapment of not only enzymes, but also organelles and microbial cells.     

天山林区六种灌木生物量的建模及其器官分配的适应性

灌木全株生物量估算模型的构建仍存在一定困难,对灌木生物量在器官分配上所体现的适应性研究也不够充分。以天山林区6种常见灌木为研究对象,在天山的东段、中段、西段林区分别设置样地进行群落调查,由此以全株收获法取得6种常见灌木若干标准株的全株、根、枝、叶及各径级根的生物量,将D~2H(地径平方与高度的乘积)与V(冠幅面积与高度的乘积)分别选为估测模型的自变量,通过回归分析法建立了各种灌木全株生物量的最优估算模型,然后比较了此6种灌木全株生物量在营养器官上分配差异以及根系生物量在径级上的分配差异。结果表明:(1)天山林区6种常见灌木中,小檗(Berberis heteropoda Schrenk)、忍冬(Lonicera hispida Pall.ex Roem.et Schuet.)、栒子(Cotoneaster melanocarpus Lodd.)的全株生物量约为8.48—9.01 kg,蔷薇(Rosa spinosissima L.)、绣线菊(Spiraea hypericifolia L.)、方枝柏(Juniperus pseudosabina Fisch.et Mey.)的全株生物量约为2.71—3.20 kg;(2)蔷薇、绣线菊、栒子的全株生物量最优估测模型是以V为自变量的函数,小檗、忍冬、方枝柏的全株生物量最优估测模型是以D~2H为自变量的函数,各模型R~2值均在0.850以上,且在P0.05水平上达到显著,模型模拟结果达到了较高的准确度;(3)6种灌木全株生物量在根、枝上的分配比重差异不显著,仅在叶上的分配比重有差异(P0.05);根系生物量在径级上的分配均呈现随根系径级下降而减少的规律,6种灌木在径级大于2 mm根上的分配比重存在差异(P0.05,径级大于20 mm根为P0.01水平);(4)6种灌木全株生物量在营养器官上的分配差异以及根系生物量在径级上的分配差异均体现了各物种对其生境选择的适应策略。     

大熊猫肠道芽孢杆菌的分离鉴定及部分生物学特性

【目的】分析大熊猫肠道中芽孢杆菌的种类、纤维素分解能力、抗微生物作用和常用抗生素药物敏感性。【方法】利用芽孢耐高温特性分离菌株,基于16S r RNA基因序列构建系统发育树,通过测量芽孢杆菌在刚果红纤维素培养基上的分解圈分析其纤维素分解能力,采用牛津杯法测定芽孢杆菌的抑菌能力,结合软件分析抑菌能力和进化树之间的关系,通过PCR调查芽孢杆菌的抗菌肽分布规律,最后通过药敏试验检测芽孢杆菌是否对常用抗生素敏感。【结果】共分离得到21株芽孢杆菌;进化树显示,这些芽孢杆菌分为6个类别(Category);羧甲基纤维素钠水解结果显示,所有菌株均能分解纤维素;大部分芽孢杆菌菌株对3种肠道病原菌有较强的抑制能力,聚类分析表明,菌株的抗菌能力与基于16S r RNA基因的分类有一定的关联性;66.67%(14/21)的菌株中可以检测到2个或3个抗菌肽基因;药敏试验结果显示,菌株整体药物耐受率低,仅为7.54%(19/264),但仍有少数菌株对抗生素耐受。【结论】分离菌株种类丰富,分布平均,且均具有纤维素分解能力。21株菌株都含有抗菌肽基因,代谢产物对3种肠道病原菌具有明显抑制作用。常用抗生素耐受性低,对规范临床用药具有指导性。     

SARS-CoV-2 envelope protein causes acute respiratory distress syndrome (ARDS)-like pathological damages and constitutes an antiviral target

Cytokine storm and multi-organ failure are the main causes of SARS-CoV-2-related death. However, the origin of excessive damages caused by SARS-CoV-2 remains largely unknown. Here we show that the SARS-CoV-2 envelope (2-E) protein alone is able to cause acute respiratory distress syndrome (ARDS)-like damages in vitro and in vivo. 2-E proteins were found to form a type of pH-sensitive cation channels in bilayer lipid membranes. As observed in SARS-CoV-2-infected cells, heterologous expression of 2-E channels induced rapid cell death in various susceptible cell types and robust secretion of cytokines and chemokines in macrophages. Intravenous administration of purified 2-E protein into mice caused ARDS-like pathological damages in lung and spleen. A dominant negative mutation lowering 2-E channel activity attenuated cell death and SARS-CoV-2 production. Newly identified channel inhibitors exhibited potent anti-SARS-CoV-2 activity and excellent cell protective activity in vitro and these activities were positively correlated with inhibition of 2-E channel. Importantly, prophylactic and therapeutic administration of the channel inhibitor effectively reduced both the viral load and secretion of inflammation cytokines in lungs of SARS-CoV-2-infected transgenic mice expressing human angiotensin-converting enzyme 2 (hACE-2). Our study supports that 2-E is a promising drug target against SARS-CoV-2.Subject terms: Cell death, Molecular biology     

Direct adenylation from 5′-OH-terminated oligonucleotides by a fusion enzyme containing Pfu RNA ligase and T4 polynucleotide kinase

5′-Adenylated oligonucleotides (AppOligos) are widely used for single-stranded DNA/RNA ligation in next-generation sequencing (NGS) applications such as microRNA (miRNA) profiling. The ligation between an AppOligo adapter and target molecules (such as miRNA) no longer requires ATP, thereby minimizing potential self-ligations and simplifying library preparation procedures. AppOligos can be produced by chemical synthesis or enzymatic modification. However, adenylation via chemical synthesis is inefficient and expensive, while enzymatic modification requires pre-phosphorylated substrate and additional purification. Here we cloned and characterized the Pfu RNA ligase encoded by the PF0353 gene in the hyperthermophilic archaea Pyrococcus furiosus. We further engineered fusion enzymes containing both Pfu RNA ligase and T4 polynucleotide kinase. One fusion enzyme, 8H-AP, was thermostable and can directly catalyze 5′-OH-terminated DNA substrates to adenylated products. The newly discovered Pfu RNA ligase and the engineered fusion enzyme may be useful tools for applications using AppOligos.     

黄土高原不同树种枯落叶混合分解效应

混交林中不同树种枯落物混合分解是否产生促进或抑制作用是评价种间关系和混交适宜性的重要依据之一。以黄土高原主要树种为对象,通过室内枯落叶混合分解模拟试验,结果表明:(1)沙棘、白榆、柠条和小叶杨枯落叶分解最快(周转期1 a左右),其次为旱柳、侧柏和白桦枯落叶(周转期略大于1 a),紫穗槐、辽东栎和刺槐枯落叶分解稍慢(周转期1.5 a左右),而樟子松、落叶松和油松枯落叶分解最慢(周转期略大于2 a)。(2)对于针叶树,与油松枯落叶混合,存在明显促进分解作用的是侧柏、落叶松,其次是白桦、沙棘和刺槐;与樟子松枯落叶混合,存在明显促进分解作用的是落叶松、侧柏、沙棘、白榆,其次是柠条、紫穗槐和小叶杨,而存在明显抑制分解作用的是刺槐,其次是白桦和辽东栎;与落叶松枯落叶混合,存在较明显促进分解作用的是白榆、白桦和辽东栎,存在较明显抑制作用的是刺槐;紫穗槐与侧柏枯落叶混合存在较明显的抑制分解作用。(3)对于阔叶树,与小叶杨枯落叶混合,存在较明显促进分解作用的是紫穗槐,其次是辽东栎和刺槐;与刺槐枯落叶混合,存在较明显促进分解作用的是白榆和沙棘,存在明显抑制分解作用的是柠条,其次是辽东栎和白桦;与白桦枯落叶混合,存在较明显促进分解作用的是辽东栎和紫穗槐,存在较明显抑制分解作用的是柠条;白榆与辽东栎、旱柳枯落叶混合均存在较明显的促进分解作用,而白榆与柠条枯落叶混合存在较明显的抑制分解作用;紫穗槐与旱柳、沙棘枯落叶混合均存在较明显的促进分解作用,而紫穗槐与柠条枯落叶混合有较明显的抑制作用。     

生态系统生态学研究的关键问题及趋势——从“整体论与还原论的争论”看

在生态学领域中,存在着生态系统整体论与还原论的争论。Tansley A.G.提出,生态系统是"准有机体"。Odum兄弟提出的"生态系统能量说"被广泛接受,但也受到质疑,称其为"还原论者的整体论"。基于对上述质疑的回应以及对生态系统整体论的追求,Patten B.C.等提出"生态网络理论",运用"网络‘环境子’分析"方法,试图从物理层面分析解决生物层面的"涌现性"问题。不过,这一理论也受到批判,认为其在探究符号化的现象对生态系统的动态影响时,陷入了还原论困境。Jrgensen S.E.等更进一步,提出"系统论"的生态系统生态学,试图从系统科学的角度研究生态系统的"物质-能量-信息-网络"系统。这一理论受到生态学界高度重视,但是也存在着在具体研究过程中如何平衡能量视角和生物地球化学视角的问题。由上述争论可见,生态系统生态学研究的趋势是从"物质实体"到"能量流动",再到"网络信息",最后到"开放系统"层层递进。目前面临的关键问题是:如何在更好地定义生态系统整体性的基础上,采取相应的能够体现生态系统整体性的方法,去获得更多、更好的生态系统整体性的认识。