Functional intracellular antibody fragments do not require invariant intra-domain disulfide bonds

Intracellular antibody fragments that interfere with molecular interactions inside cells are valuable in investigation of interactomes and in therapeutics, but their application demands that they function in the reducing cellular milieu. We show here a 2.7-Å crystal structure of intracellular antibody folds based on scaffolds developed from intracellular antibody capture technology, and we reveal that there is no structural or functional difference with or without the intra-domain disulfide bond of the variable domain of heavy chain or the variable domain of light chain. The data indicate that, in the reducing in vivo environment, the absence of the intra-domain disulfide bond is not an impediment to correction of antibody folding or to interaction with antigen. Thus, the structural constraints for in-cell function are intrinsic to variable single-domain framework sequences, providing a generic scaffold for isolation of functional intracellular antibody single domains.     

Lung Cancer: Are we up to the Challenge?

Lung cancer is the leading cause of cancer deaths worldwide among both men and women, with more than 1 million deaths annually. Non-small cell lung cancer (NSCLC) accounts for about 80% of all lung cancers.Although recent advances have been made in diagnosis and treatment strategies, the prognosis of NSCLC patients is poor and it is basically due to a lack of early diagnostic tools.However, in the last years genetic and biochemical studies have provided more information about the protein and gene's mutations involved in lung tumors. Additionally, recent proteomic and microRNA's approaches have been introduced to help biomarker discovery.Here we would like to discuss the most recent discoveries in lung cancer pathways, focusing on the genetic and epigenetic factors that play a crucial role in malignant cell proliferation, and how they could be helpful in diagnosis and targeted therapy.     

大鼠下丘脑室旁核与终纹床核及前额叶皮质间纤维联系的研究

分别注射辣根过氧化物酶（HRP）入大鼠的PVN和BNST,用组织化学的方法在确定注射部位准确的情况下,在PVN、BNST及PFC观察被标记的神经元或轴突末梢,探讨大鼠下丘脑室旁核（PVN）与终纹床核（BNST）及前额叶皮质间（PFC）之间是否存在投射通路;将HRP注射到PVN后,在同侧的BNST见标记的细胞体,在PFC未见标记的细胞体或轴突末梢;将HRP注射到BNST后,在同侧的PVN见标记的轴突末梢,在PFC未见标记的细胞体或轴突末梢。大鼠BNST有神经纤维投射到PVN,PFC与PVN及BNST之间没有直接的或只有极少量的纤维联系,在机体面临威胁性情境时,BNST可能激活HPA轴引发生理和行为反应,PFC是否通过与PVN或BNST的直接或间接的纤维投射实现其调节功能值得关注。     

甲氰菊酯和辛硫磷及其混剂的土壤微生物降解

The degradation rates of fenpropathrin, phoxim and their mixture in un-sterilized soil were much quicker-than those in sterilized soil, which indicated that soil microorganisms played a significant role in the degradation pro-cessin soil. The half-life (T0.5) in un-sterilized soil was 56.2 d for fenpropathrin, 57.8 d for mixed fen-propathrin, 48.2 d for phoxim, and 41.7 d for mixed phoxim. The corresponding half-life (T0.5) in sterilizedsoil was 135.1 d, 147.3 d, 123.6 d, and 126.2 d, respectively. There were no significant differences for degradation rates between single use and mixed use of fenpropathrin and nhoxirn.     

Primary structure of apolipophorin-III from the greater wax moth,Galleria mellonella

The complete amino acid sequence of apolipophorin-III (apoLp-III), a lipid-binding hemolymph protein from the greater wax moth,Galleria mellonella, was determined by protein sequencing. The mature protein consists of 163 amino acid residues forming a protein of 18,075.5 Da. Its sequence is similar to apoLp-III from other Lepidopteran species, but remarkably different from the apoLp-IIIs of insects from other orders. As shown by mass spectrometric analysis, the protein carries no modifications. Thus, all of its known physiological functions, including its recently discovered immune response-stimulating activity, must reside in the protein itself.     

Photoinhibition of photosynthesis represents a mechanism for the long-term regulation of photosystem II

The obligate shade plant, Tradescantia albiflora Kunth grown at 50 mol photons · m^–2 s^–1 and Pisum sativum L. acclimated to two photon fluence rates, 50 and 300 mol · m^–2 · s^–1, were exposed to photoinhibitory light conditions of 1700 mol · m^–2 · s^–1 for 4 h at 22° C. Photosynthesis was assayed by measurement of CO₂-saturated O₂ evolution, and photosystem II (PSII) was assayed using modulated chlorophyll fluorescence and flash-yield determinations of functional reaction centres. Tradescantia was most sensitive to photoinhibition, while pea grown at 300 mol · m^–2 · s^–1 was most resistant, with pea grown at 50 mol · m^–2 · s^–1 showing an intermediate sensitivity. A very good correlation was found between the decrease of functional PSII reaction centres and both the inhibition of photosynthesis and PSII photochemistry. Photoinhibition caused a decline in the maximum quantum yield for PSII electron transport as determined by the product of photochemical quenching (q_p) and the yield of open PSII reaction centres as given by the steady-state fluorescence ratio, F_vF_m, according to Genty et al. (1989, Biochim. Biophys. Acta 990, 81–92). The decrease in the quantum yield for PSII electron transport was fully accounted for by a decrease in F_vF_m, since q_p at a given photon fluence rate was similar for photoinhibited and noninhibited plants. Under lightsaturating conditions, the quantum yield of PSII electron transport was similar in photoinhibited and noninhibited plants. The data give support for the view that photoinhibition of the reaction centres of PSII represents a stable, long-term, down-regulation of photochemistry, which occurs in plants under sustained high-light conditions, and replaces part of the regulation usually exerted by the transthylakoid pH gradient. Furthermore, by investigating the susceptibility of differently lightacclimated sun and shade species to photoinhibition in relation to q_p, i.e. the fraction of open-to-closed PSII reaction centres, we also show that irrespective of light acclimation, plants become susceptible to photoinhibition when the majority of their PSII reaction centres are still open (i.e. primary quinone acceptor oxidized). Photoinhibition appears to be an unavoidable consequence of PSII function when light causes sustained closure of more than 40% of PSII reaction centres.Abbreviations F_o and F_o minimal fluorescence when all PSII reaction centres are open in darkness and steady-state light, respectively - F_m and F_m maximal fluorescence when all PSII reaction centres are closed in darkand light-acclimated leaves, respectively - F_v variable fluorescence - (F_m-F_o) under steady-state light con-ditions - F_s steady-state fluorescence in light - Q_A the primary,stable quinone acceptor of PSII - q_Ne non-photochemical quench-ing of fluorescence due to high energy state - (pH); q_Ni non-photochemical quenching of fluorescence due to photoinhibition - q_p photochemical quenching of fluorescence To whom correspondence should be addressedThis work was supported by the Swedish Natural Science Research Council (G.Ö.) and the award of a National Research Fellowship to J.M.A and W.S.C. We thank Dr. Paul Kriedemann, Division of Forestry and Forest Products, CSIRO, Canberra, Australia, for helpful discussions.     

甘肃金水湖国家湿地公园景观自然性的时空异质性

景观自然性体现了各景观要素的时空分布格局,对提高生态系统的整体性和稳定性有重要意义.通过解译2003年、2009年和2015年3个时相的遥感影像,利用GIS和层次分析法,研究了甘肃金川金水湖国家湿地公园景观自然性变化规律.结果表明:随着时间的变化,景观结构中景观多样性指数、均匀度指数、斑块丰富度呈增加趋势,景观优势度、景观分离度呈减小趋势;提供栖息地、植物群落维持功能、面积加权平均斑块分维数、植被覆盖度等生态功能指标呈增大趋势;人工干扰指数、人工景观平均面积等生态学干扰指标呈减小趋势,湿地公园的景观自然性指数由2003年的0.4761增加至2015年的0.7485.湿地公园保育和恢复工程的实施改变了土地覆被结构,增强了生态系统的稳定性、生态弹性和服务价值,较大程度上提高了金水湖国家湿地公园的景观自然性.     

安徽部分湖泊养殖鱼类暴发性出血病的病原分离与鉴定

目的确定引起安徽省部分湖泊养殖鱼类暴发性出血病的病原,为防治该病提供理论依据。方法随机取濒死期鲫鱼和白鲢的肌肉组织分别进行细菌和病毒分离培养,联合采用细菌表型鉴定法和16S rRNA基因序列分析法鉴定分离菌株,并使用分离菌株进行人工感染实验。结果从5尾患病鱼的肌肉组织中分离获得5株细菌,综合分离菌株的形态特征、理化特性和16S rRNA基因序列与系统发育学分析的结果,确定L1菌株为温和气单胞菌、L2菌株为维氏气单胞菌、J1、J2和L3菌株为嗜水气单胞菌。人工感染实验表明这5个气单胞菌分离株均具有较强的致病性。结论气单胞菌是该次鱼类暴发性出血病的病原,水温剧变、水质恶化和缺氧是疾病暴发的诱因,应采取改善环境、加强饲养管理、提高鱼体抵抗病力和及时杀灭病原体的综合措施防控该病。     

测定胸腺肽含量几种定量方法的比较

本文用凯氏定氮法、双缩脲法及福林－酚法测定了胸腺肽含量,并加以比较,说明三种方法对其含量测定没有显著差异．在实践中可以根据不同条件选择适当的方法进行实验．     

Cyclic biphalin analogues with a novel linker lead to potent agonist activities at mu,delta, and kappa opioid receptors

In an effort to improve biphalin’s potency and efficacy at the µ-(MOR) and δ-opioid receptors (DOR), a series of cyclic biphalin analogues 1–5 with a cystamine or piperazine linker at the C-terminus were designed and synthesized by solution phase synthesis using Boc-chemistry. Interestingly, all of the analogues showed balanced opioid agonist activities at all opioid receptor subtypes due to enhanced κ-opioid receptor (KOR) activity. Our results indicate that C-terminal flexible linkers play an important role in KOR activity compared to that of the other cyclic biphalin analogues with a hydrazine linker. Among them, analogue 5 is a potent (Ki?=?0.27, 0.46, and 0.87?nM; EC₅₀?=?3.47, 1.45, and 13.5?nM at MOR, DOR, and KOR, respectively) opioid agonist with high efficacy. Based on the high potency and efficacy at the three opioid receptor subtypes, the ligand is expected to have a potential synergistic effect on relieving pain and further studies including in vivo tests are worthwhile.