饲料中狐狸、水貂、貉子和狗源性的五重实时荧光PCR检测方法的建立

为了快速鉴别饲料中的狐狸、水貂、貉子和狗源性成分,根据线粒体16S r DNA种间保守序列,设计合成针对狐狸、水貂、貉子和狗的特异性引物和探针,通过对荧光PCR反应体系和反应条件的优化筛选,建立了多重实时荧光PCR方法,在同一PCR反应体系中可以同时完成4种动物源性成分检测。通过对15种其他物种的源性成分的检测,结果表明所设计的引物和探针具有很好的物种特异性,且灵敏度高,狐狸、水貂、貉子和狗的DNA检出限为0.01ng。对40份样品检测,其中5份检测出貉子、狐狸和水貂源性成分。结果表明,该方法可以有效地鉴别出饲料中狐狸、水貂、貉子和狗源性成分,同时适用于相关动物产品中。     

太白山巴山冷杉林主要树种与开花秦岭箭竹的空间点格局分析

采用单变量和双变量O-ring函数对太白山大熊猫栖息地巴山冷杉林主要树种的空间分布格局、种间关联性及其与林下开花箭竹的空间关联性进行了多尺度分析.结果表明: 巴山冷杉林中,巴山冷杉数量最多,但种群结构衰退,白桦种群相对年轻,种群结构稳定,红桦种群也呈衰退趋势;3个主要树种在小尺度上呈聚集分布,随尺度增加,逐渐表现为随机分布.3个树种的空间关联性主要表现在小尺度范围内,随尺度增加,空间分布格局逐渐表现为不关联;巴山冷杉和白桦与开花秦岭箭竹在大、中尺度内呈现正相关,而红桦与开花秦岭箭竹在大、中尺度上表现出负相关.大熊猫栖息地内乔木和林下秦岭箭竹共同推动森林的动态演替和发展,进而影响秦岭大熊猫栖息地的环境变化.     

四川省汶川县生态经济系统能值分析

基于能值理论和分析方法,应用统计和实地调查数据,对四川省汶川县1982-2002年的生态经济系统进行了分析.结果表明:汶川县的经济水平较低,规模较小;主要能值投入为可再生资源;能值产出中不可再生资源能值占大部分,主要来自矿产资源和水泥生产;研究区输出大量能值,项目包括矿产资源和木材资源,大量的能值输出,特别是木材能值输出已使得研究区脆弱的生态环境出现生态问题;汶川县的人均能值消费水平较低,生活水平不高;研究区应注意经济与生态的协调发展.     

In Vitro Intracellular Trafficking of Virulence Antigen during Infection by Yersinia pestis

Yersinia pestis, the causative agent of plague, encodes several essential virulence factors on a 70 kb plasmid, including the Yersinia outer proteins (Yops) and a multifunctional virulence antigen (V). V is uniquely able to inhibit the host immune response; aid in the expression, secretion, and injection of the cytotoxic Yops via a type III secretion system (T3SS)-dependent mechanism; be secreted extracellularly; and enter the host cell by a T3SS-independent mechanism, where its activity is unknown. To elucidate the intracellular trafficking and target(s) of V, time-course experiments were performed with macrophages (MΦs) infected with Y. pestis or Y. pseudotuberculosis at intervals from 5 min to 6 h. The trafficking pattern was discerned from results of parallel microscopy, immunoblotting, and flow cytometry experiments. The MΦs were incubated with fluorescent or gold conjugated primary or secondary anti-V (antibodies [Abs]) in conjunction with organelle-associated Abs or dyes. The samples were observed for co-localization by immuno-fluorescence and electron microscopy. For fractionation studies, uninfected and infected MΦs were lysed and subjected to density gradient centrifugation coupled with immunoblotting with Abs to V or to organelles. Samples were also analyzed by flow cytometry after lysis and dual-staining with anti-V and anti-organelle Abs. Our findings indicate a co-localization of V with (1) endosomal proteins between 10–45 min of infection, (2) lysosomal protein(s) between 1–2 h of infection, (3) mitochondrial proteins between 2.5–3 h infection, and (4) Golgi protein(s) between 4–6 h of infection. Further studies are being performed to determine the specific intracellular interactions and role in pathogenesis of intracellularly localized V.     

DEPOLARIZING STIMULI AND THE RELEASE OF PHYSIOLOGICALLY ACTIVE AMINO ACIDS FROM SUSPENSIONS OF MAMMALIAN SYNAPTOSOMES

—Potassium causes increased metabolism and release of physiologically active amino acids from suspensions of mammalian synaptosomes. Sheep and rat hypothalamic synaptosomes, and rabbit, sheep and rat cortical synaptosomes show respiratory and glycolytic response to electrical stimulation and show a parallel calcium-dependent differential release of physiologically active amino acids. Attempts were made to eliminate the actions of heating and electrode products as the agents causing the observed effects and the case is made that depolarization is the trigger mechanism.     

High SEPT9_v1 Expression Is Associated with Poor Clinical Outcomes in Head and Neck Squamous Cell Carcinoma

The purpose of this work was to determine SEPT9_v1 expression levels in head and neck squamous cell carcinoma (HNSCC) and to analyze whether SEPT9_v1 expression is relevant to clinical outcomes. Recently, the SEPT9 isoform SEPT9_v1 has been implicated in oncogenesis, and methylation of the SEPT9 promoter region was reported in HNSCC. These findings led us to hypothesize that SEPT9_v1 could be differently expressed in HNSCC. To determine whether SEPT9_v1 is expressed in HNSCC, tissue microarray immunohistochemical analysis was performed using a SEPT9_v1-specific antibody. Tissue microarrays stained with a polyclonal SEPT9_v1-specific antibody was used to determine protein expression levels in HNSCC tissue samples, some with known clinical outcomes. This analysis showed that SEPT9_v1 is in fact highly expressed in HNSCC compared with normal epithelium, and high expression levels directly correlated with poor clinical outcomes. Specifically, a high SEPT9_v1 expression was associated with decreased disease-specific survival (P = .012), time to indication of surgery at primary site (P = .008), response to induction chemotherapy (P = .0002), and response to chemotherapy (P = .02), as well as advanced tumor stage (P = .012) and N stage (P = .0014). The expression of SEPT9_v1 was also strongly correlated with smoking status (P = .00094). SEPT9_v1 is highly expressed in HNSCC, and a high expression of SEPT9_v1 is associated with poor clinical outcomes. These data indicate that SEPT9_v1 warrants additional investigation as a potential biomarker for HNSCC.     

Amino acid sequence surrounding the lipoic acid cofactor of bovine kidney 2-oxoglutarate dehydrogenase complex

The 2-oxoglutarate dehydrogenase complex was succinylated using 2-oxo[5-14C]glutarate in the presence of N-ethylmaleimide to label the lipoic acid cofactor of the transuccinylase (E2) component. Following peptic digestion, 14C-lipoate-containing peptides were purified and subjected to automated Edman degradation and amino acid analysis. The amino acid sequence surrounding the lipoyllysine residue is reported.