A deep-red xanthene-based highly sensitive fluorescent probe for detection of hypochlorite

As an oxidant, deodorant and bleaching agent, the hypochlorous acid (HClO) and hypochlorite (ClO⁻) are widely used in corrosion inhibitors, textile dyes, pharmaceutical intermediates and in our daily lives. However, excess usage or aberrant accumulation of ClO⁻ leads to tissue damage or some diseases and even cancer. Therefore, it is necessary to develop a fluorescent probe that specifically identifies ClO⁻. In this article, we synthesized a deep-red xanthene-based fluorescent probe (XA-CN). The strong electron deficient group dicyano endows the probe XA-CN deep-red fluorescent emission with high solubility, selectivity and sensitivity for ClO⁻ detection. Studies showed that the probe demonstrated turn-off fluorescence (643 nm) at the presence of ClO⁻ in dimethylsulfoxide/phosphate-buffered saline 1:1 (pH 9) solution with a limit of detection of 1.64 μM. Detection mechanism investigation revealed that the electron deficient group -CN and the hydroxyl group was oxidized into aldehyde or carbonyl groups at the presence of ClO⁻, resulting ultraviolet-visible absorption of the probe blue shifted and turned-off fluorescence. Furthermore, XA-CN was successfully used for the detection of ClO⁻ in tap water samples.     

外源胆固醇对水稻幼苗膜脂组成及抗冷力的影响

分析了水稻幼苗低温胁迫前后膜脂和膜脂脂肪酸含量变化。结果表明,经胆固醇处理的幼苗叶和根细胞膜脂中LPC、PS和PG含量比对照下降少,PA含量增加也较少。胆固醇处理的幼苗叶和根棕榈酸(16:0)增加量和亚麻酸(18:3)与IUFA减少量均明显比对照少。试验结果证明,水稻幼苗叶片和根系的抗冷力与PA含量和脂肪酸不饱和程度变化有密切关系。外源胆固醇处理水稻幼苗能阻止低温对膜脂的破坏作用,提高幼苗抗低温胁迫能力。     

河南省伏牛山南侧西峡县的森林植被及其合理利用问题

本文是在野外调查的基础上,参考有关资料和科研成果写成的,对于河南伏牛山南侧西峡县森林植被的生态环境、主要类型及其分布、与生态环境的关系、动态变化以及合理利用问题作初步论述。     

Stratified Communities of Methanogens in the Jiulong River Estuarine Sediments, Southern China

Methane is a potent greenhouse gas and produced mainly by methanogens. Few studies have specifically dealt so far with methanogens in estuarine environments. In this study, diversity and distribution of methanogens were investigated by clone library and T-RFLP analysis in a Jiulong River estuarine sediment core which contained clear sulfate–methane-transition zone. The majority of obtained sequences in clone libraries and T-RF peaks from T-RFLP analysis were assigned mainly to Methanosaeta, Methanomicrobiales and Methanosarcinales/ANME. The fragments of Methanosarcinales/ANME were most dominant group (mean 51 %) and composed largely of ANME-2a. In addition, Methanosaeta and Methanomicrobiales accounted for 21 and 28 % of all fragments. Therefore, the presence of Methanomicrobiales, Methanosaeta and ANME-2a was indicative of acetoclastic methanogenesis, hydrogenotrophic methanogenesis, and anaerobic methane oxidation in Jiulong River estuarine sediments. This study provided the important knowledge towards understanding methane cycling association of representative of methanogens involved in estuarine environments.     

Structural Basis for the Dual Recognition of IL-12 and IL-23 by Ustekinumab

Interleukin (IL)-12 and IL-23 are heterodimeric proinflammatory cytokines that share a common p40 subunit, paired with p35 and p19 subunits, respectively. They represent an attractive class of therapeutic targets for the treatment of psoriasis and other immune-mediated diseases. Ustekinumab is a fully human monoclonal antibody (mAb) that binds specifically to IL-12/IL-23p40 and neutralizes human IL-12 and IL-23 bioactivity. The crystal structure of ustekinumab Fab (antigen binding fragment of mAb), in complex with human IL-12, has been determined by X-ray crystallography at 3.0 Å resolution. Ustekinumab Fab binds the D1 domain of the p40 subunit in a 1:1 ratio in the crystal, consistent with a 2 cytokines:1 mAb stoichiometry, as measured by isothermal titration calorimetry. The structure indicates that ustekinumab binds to the same epitope on p40 in both IL-12 and IL-23 with identical interactions. Mutational analyses confirm that several residues identified in the IL-12/IL-23p40 epitope provide important molecular binding interactions with ustekinumab. The electrostatic complementarity between the mAb antigen binding site and the p40 D1 domain epitope appears to play a key role in antibody/antigen recognition specificity. Interestingly, this structure also reveals significant structural differences in the p35 subunit and p35/p40 interface, compared with the published crystal structure of human IL-12, suggesting unusual and potentially functionally relevant structural flexibility of p35, as well as p40/p35 recognition. Collectively, these data describe unique observations about IL-12p35 and ustekinumab interactions with p40 that account for its dual binding and neutralization of IL-12 and IL-23.     

阳宗海和滇池中紫色非硫细菌数量和种群结构的比较

在2002年平水期对高原湖泊阳宗海和滇池中的紫色非硫细菌(PNSB)数量和种群结构进行了比较研究。对两湖中的各因子进行t检验,发现各因子和PNSB数量都有极显著差异,富营养化湖泊滇池中的PNSB数量远远多于贫营养化湖泊阳宗海;两湖中PNSB的优势种是红假单胞菌属,也有少数的红螺菌属。两湖种群结构的区别是滇池有红微菌,而阳宗海有大量红细菌。     

Expression profile of human immune-responsive gene 1 and generation and characterization of polyclonal antiserum

Murine immune-responsive gene 1 (IRG1) plays significant roles in embryonic implantation and neurodegeneration. The expression pattern of the human IRG1 gene, however, has not yet been established, and the predicted gene sequence has been revised several times according to computed expressed sequence tags (ESTs). To determine the human IRG1 gene expression profile, human fetal tissue samples, peripheral blood mononuclear cells (PBMCs) from normal healthy subjects, and the human leukemia cell lines THP-1 and K-562 challenged with lipopolysaccharide (LPS) were subjected to RT-PCR using degenerate primers. The results indicated that the IRG1 gene is differentially expressed in human fetal PBMCs and LPS-stimulated adult PBMCs. The amplified gene fragment was cloned into the pET32a(+) vector and fusion-expressed with a His-tag in a prokaryotic system. After affinity chromatography, human IRG1h fusion proteins were isolated by SDS-PAGE and identified by mass spectrometric analysis for use as an immunogen to immunize rabbits. The titer and specificity of the purified rabbit antiserum were sufficient to measure human IRG1 gene expression in various tissues and cultures. This purified polyclonal antiserum will allow us to initiate studies to elucidate the biological roles of the human IRG1 gene.     

Generation and evaluation of anti-mouse IgG IgY as secondary antibody

Abstract

In order to evaluate the possibility of using IgY as the secondary antibody in immunoassay, specific IgY (1: 128,000) was generated by immunizing hens with mouse serum IgG purified by protein A column. IgY was extracted from egg yolk by polyethylene glycol 6000 (PEG-6000), and further purified using protein M affinity chromatography column. The purified IgY was conjugated with horseradish peroxidase (HRP) and fluorescein?isothiocyanate (FITC), in that order. The reactivity of conjugated antibodies was evaluated by ELISA, Western blot and Immunofluorescence, demonstrating that the obtained IgY was able to conjugate with enzymes, react with mouse primary IgG antibody, and subsequently amplify the antigen-antibody signals in different immune reaction conditions, in a comparable secondary effect to conventional goat anti-mouse IgG antibody. The obtained conjugated antibodies showed high stability in broad pH ranges (4–10; >70%) and high thermostability at 37?°C for 84?h (>85%). Despite the need to further consider and evaluate the industrial standardization and production process, our data provided the primary evidence that conjugated IgY antibodies can be used as a secondary antibody for broad immunological analysis.