Identification of the β-Lactamase Inhibitor Protein-II (BLIP-II) Interface Residues Essential for Binding Affinity and Specificity for Class A β-Lactamases

The interactions between β-lactamase inhibitory proteins (BLIPs) and β-lactamases have been used as model systems to understand the principles of affinity and specificity in protein-protein interactions. The most extensively studied tight binding inhibitor, BLIP, has been characterized with respect to amino acid determinants of affinity and specificity for binding β-lactamases. BLIP-II, however, shares no sequence or structural homology to BLIP and is a femtomolar to picomolar potency inhibitor, and the amino acid determinants of binding affinity and specificity are unknown. In this study, alanine scanning mutagenesis was used in combination with determinations of on and off rates for each mutant to define the contribution of residues on the BLIP-II binding surface to both affinity and specificity toward four β-lactamases of diverse sequence. The residues making the largest contribution to binding energy are heavily biased toward aromatic amino acids near the center of the binding surface. In addition, substitutions that reduce binding energy do so by increasing off rates without impacting on rates. Also, residues with large contributions to binding energy generally exhibit low temperature factors in the structures of complexes. Finally, with the exception of D206A, BLIP-II alanine substitutions exhibit a similar trend of effect for all β-lactamases, i.e., a substitution that reduces affinity for one β-lactamase usually reduces affinity for all β-lactamases tested.     

基于SBE法的广东省生态景观林带美景度评价

采用美景度评价法(SBE法)对广东省生态景观林带进行评价，分析影响生态景观林带景观水平的主要因子，探讨生态景观林带景观构成要素与景观质量的关系。结果表明，不同专业、性别、学历、年龄的人群在森林审美态度上具有一致性，其评判结果能够反映森林美景度的实际情况；对53张照片评价SBE值，最大为1.4454，最小为-1.6917；对美景度影响较大的4个因子，即色彩丰富度(X6)、色泽明度(X7)、生活型(X8)、生长状况(X9)，建立的多元线性回归模型是 Y=1.902-0.346X6-0.461X7+ 0.206X8-0.584X9。     

贵州百里杜鹃风景区旅游干扰对杜鹃群落特征的影响

为揭示旅游干扰对于百里杜鹃风景区杜鹃群落特征影响,选取杜鹃纯林和杜鹃混交林两种杜鹃群落,分析干扰对群落结构及物种多样性的影响。采用典型抽样的方法进行样地设置和调查,通过旅游影响系数分析样地旅游干扰等级,并分析对比了不同旅游干扰程度和干扰方式对群落特征的影响。研究表明：1）百里杜鹃景区共有植被67科102属130种,其中杜鹃纯林共有植被97种,以马缨杜鹃、箭竹和露珠杜鹃为优势物种,杜鹃混交林共有植被115种,以马缨杜鹃、露珠杜鹃和映山红为优势种;2）两种杜鹃群落均在中等干扰水平时具有最高的Berger-Parker丰富度指数和Shannon多样性指数,中度干扰一定程度上促进了杜鹃群落物种多样性增加,不同的是两种群落的Shannon指数变化趋势不同,在杜鹃纯林中表现为中度干扰>轻度干扰>重度干扰,而在杜鹃混交林中中度干扰>重度干扰>轻度干扰,相较于杜鹃纯林,杜鹃混交林物种更为丰富,在旅游干扰下生物多样性更为稳定,抗干扰能力更强,而杜鹃纯林受旅游干扰波动显著;3）垃圾指数、踩踏指数与杜鹃混交林物种多样性指数、群落生长结构无明显相关性,而垃圾指数与杜鹃纯林Shannon多样性指数、物种数以及草本均高呈显著负相关,踩踏指数与杜鹃纯林灌木层高度、盖度呈现极显著负相关,与草本均高呈显著正相关;4）各杜鹃群落对旅游干扰有不同响应,为保持杜鹃群落稳定性,应在花季适当分流,严格控制杜鹃纯林的重度干扰。     

Sinking rates of late-stage fish larvae: Implications for larval ingress into estuarine nursery habitats

Many species spawn in oceanic waters yet their juvenile stages use nearshore and estuarine habits and the bio-physical mechanisms by which late-larvae enter these juvenile habitats may be an important bottleneck in the population dynamics of these species. To provide parameters for the development of larval ingress models, sinking rates were measured of the late-stage larvae of six fish species: Atlantic croaker (Micropogonias undulatus), spot (Leiostomus xanthurus), Atlantic menhaden (Brevoortia tyrannus), summer flounder (Paralichthys dentatus), southern flounder (P. lethostigma), and gulf flounder (P. albigutta). Species-specific differences were found in sinking rates; Atlantic croaker had the slowest sinking rates and Atlantic menhaden and the three flounder species had the fastest sinking rates. Additionally, sinking rates increased for all species as length increased. The total amount of variability explained in sinking rates was low (20–50%), indicating a large amount of variability at the level of the individual. The observed patterns in sinking rates were then combined with previous studies on the mechanisms of larval ingress to present species-specific conceptual models of ingress.     

抗对虾白斑综合症病毒的单链抗体A1的表达和生物化学特性

用噬菌体展示技术制备了抗对虾白斑综合症病毒(WSSV)的单链抗体A1。该抗体在30℃培养条件下诱导表达20h后,其蛋白表达量可达总菌体蛋白的3.67%。用亲和层析柱和SephadexG-100层析柱可将单链抗体A1纯化为一条单电泳条带,其分子量约为31.5kD。用等电聚焦电泳测定,其等电点为pH5.8。ELISA测定表明冻干的单链抗体A1在室温储藏4年后与WSSV结合仍具有较高的活力。     

Agrobacterium-mediated transformation of Guignardia citricarpa: An efficient tool to gene transfer and random mutagenesis

Guignardia citricarpa is the causal agent of Citrus Black Spot (CBS), an important disease in Citriculture. Due to the expressive value of this activity worldwide, especially in Brazil, understanding more about the functioning of this fungus is of utmost relevance, making possible the elucidation of its infection mechanisms, and providing tools to control CBS. This work describes for the first time an efficient and successful methodology for genetic transformation of G. citricarpa mycelia, which generated transformants expressing the gene encoding for the gfp (green fluorescent protein) and also their interaction with citrus plant. Mycelia of G. citricarpa were transformed via Agrobacterium tumefaciens, which carried the plasmid pFAT-gfp, contains the genes for hygromycin resistance (hph) as well as gfp. The optimization of the agrotransformation protocol was performed testing different conditions (type of membrane; inductor agent concentration [acetosyringone – AS] and cocultivation time). Results demonstrated that the best condition occurred with the utilization of cellulose's ester membrane; 200 μM of AS and 96 h as cocultivation time. High mitotic stability (82 %) was displayed by transformants using Polymerase Chain Reaction (PCR) technique to confirm the hph gene insertion. In addition, the presence of gfp was observed inside mycelia by epifluorescence optical microscopy. This technique easy visualization of the behaviour of the pathogen interacting with the plant for the first time, allowing future studies on the pathogenesis of this fungus. The establishment of a transformation method for G. citricarpa opens a range of possibilities and facilitates the study of insertional mutagenesis and genetic knockouts, in order to identify the most important genes involved in the pathogenesis mechanisms and plant–pathogen interaction.     

Fungitoxic Effects of Extracts of Azadirachta Indica against Cochliobolus Miyabeanus causing Brown Spot Disease of Rice

The oil extract from seeds of Azadirachta indica as well as water and ethanol leaf extracts of the plant were effective in reducing the radial growth of Cochliobolus miyabeanus in culture and in checking the spread of brown spot disease in rice. The oil extract was found to be the most effective, followed by ethanol leaf extract, in inhibiting the growth of the pathogen in vitro and in checking the development of the disease in vivo . Although the cold water extract from the plant leaf reduced the radial growth of the pathogen in culture, it failed to reduce the incidence and severity of the disease as effectively as the oil and ethanol extracts. The oil and ethanol extracts compared favourably with carbendazim (Bavistin) at 0.1% a.i and have the potential for control of brown spot disease of rice in the field.     

英德西南旅游区水源河流水质与浮游生物群落特征

通过野外采样及实验室分析,调查了英德市西南旅游区水源河流上、中、下游的水质状况及浮游生物的多样性。结果表明,(1)英德市西南旅游区上、中、下游河段亚硝酸盐、氨、硝酸盐、总氮、镉、铅、铬、镍、铜、锌的含量低于国家标准,铝、钙、钴、铁、钼、硒的含量超过国家标准,而总磷、砷、镁、锰的含量在部份区段已超过国家标准;(2)浮游动物种类的个体数在整条河中以桡足类最多,上游通天岩中浮游动物个体数最多,下游碧落湖浮游动物多样性最低;此次调查总共发现68种浮游植物,其中主要集中在下游碧落湖,在中游碧落洞绿藻、蓝藻、硅藻总个体数最低,上游通天岩绿藻及甲藻总个体数最多,而下游碧落湖的蓝藻和硅藻的总个体数最多：（3）总氮、总磷、铅、钙、浮游动物Shannon-Weaver指数与浮游植物Shannon-Weaver指数从上游向下游增加,而锰与镍含量则成相反趋势。文中探讨了水中元素之间、元素与生物之间、浮游动植物之间的相互关系。本文对风景区水源的生态环境的研究风景区旅游资源开发与管理提供参考。     

一种筛选对虾白斑综合症病毒中和抗体的新方法

用对虾淋巴组织的原代培养细胞与对虾白斑综合症病毒进行相互作用,洗去不能结合的病毒,然后用甲醛固定,再用ELISA法检测所吸附的病毒。当病毒与抗血清保温后,抑制病毒与细胞的吸附能力的抗血清中应含有中和抗体。此方法可以在缺少细胞的情况下,简便地筛选到中和抗体。