A snake venom metalloproteinase that inhibited cell proliferation and induced morphological changes of ECV304 cells

TSV-DM, a basic metalloproteinase with a molecular weight of 110 kDa, was purified from Trimeresurus stejnegeri venom. TSV-DM degraded the A alpha chain of fibrinogen more rapidly than the B beta chain in a dose dependent manner. The cDNA of TSV-DM encode

竹叶青蛇毒金属蛋白酶结构与功能的研究

出血性的蝰科蛇毒中含有丰富的蛇毒金属蛋白酶，本论文就竹叶青（Trimeresurus stejnegeri）蛇毒中的蛇毒金属蛋白酶的结构与功能进行研究。我们用生物化学手段从竹叶青（T.stejnegeri）的粗毒中分离纯化得到一个二聚体的P-IIIb亚型蛇毒金属蛋白酶，命名为TSV-DM。同时，用分子生物学方法从竹叶青（T.stejnegeri）的毒腺cDNA文库中克隆得到3个P-III型的蛇毒金属蛋白酶的cDNAs序列， 其中一个编码TSV-DM蛋白前体，另两个编码P-IIIc亚型的出血性蛇毒金属蛋白酶前体，分别命名为stejnihagin-A 和 stejnihagin-B。 经过阴离子层析和肝素亲和层析两步层析方法，我们从竹叶青（T. stejnegeri）蛇毒中分离纯化得到TSV-DM蛋白质，非还原条件下SDS-PAGE电泳表观分子量约110 kDa，还原条件下约为55 kDa。活性检测表明，TSV-DM降解牛纤维蛋白原Aα链快于Bβ链，且呈剂量依赖关系。但不降解明胶，不诱导出血，不具有促凝或者抗凝活性，以及不诱导或者抑制血小板聚集。蛋白质N-末端测序表明TSV-DM的成熟蛋白N-末端封闭。利用肽指纹图谱确证了TSV-DM的编码cDNA。TSV-DM的cDNA序列编码622个氨基酸残基的蛋白前体，包括信号肽、前肽、金属蛋白酶区域、间隔区、去整合素样区域和富含半胱氨酸区域。TSV-DM与其他P-III型蛇毒金属蛋白酶的一级结构序列比对发现TSV-DM和诱导血管内皮细胞凋亡的P-IIIb亚型蛇毒金属蛋白酶具有高度的同源性。但是用人脐带静脉血管内皮细胞系ECV304细胞作为靶细胞检测TSV-DM的诱导血管内皮细胞凋亡活性发现，TSV-DM只能抑制ECV304细胞的增殖和诱导细胞形态从多角形的内皮细胞向成纤维细胞样的梭形状改变。电泳检测抽提的片断化DNA以及流式细胞仪检测TSV-DM处理的ECV304细胞的DNA含量变化均表明TSV-DM不能诱导ECV304细胞的凋亡。 Stejnihagin-A 和stejnihagin-B是用PCR方法从竹叶青（T.stejnegeri）毒腺cDNA文库中克隆得到的两个P-III型蛇毒金属蛋白酶前体的cDNAs。这两个cDNA序列均编码600个氨基酸残基的蛋白前体，包括信号肽、前肽、金属蛋白酶区域、间隔区、去整合素样区域和富含半胱氨酸区域。推导成熟肽的氨基酸序列分析结果表明，stejnihagin-A 和stejnihagin-B不仅在一级结构序列上和来源于黄绿烙铁头（Trimeresurus flavoviridis）的HR1b具有高度同源性，高达79%， 而且在他们的金属蛋白酶区域的第100个氨基酸残基位置上均有一保守的半胱氨酸残基。结合序列比对和进化树的分析，我们推测stejnihagin-A、stejnihagin-B和HR1b有可能组成一个新的P-III型蛇毒金属蛋白酶亚型，命名为P-IIIc亚型。

Tobacco Streak Virus (TSV) in Cotton - scoping study

This project aims to examine the possible impact of Tobacco Streak Virus (TSV) on the Australian cotton industry. TSV is transmitted by thrips, causes a disease which has had a significant impact on grain crops in Central Queensland and a preliminary study in 2007 has shown that cotton is also susceptible to field infection in this region, but many questions remain unanswered. This project aims to: • Determine the impact of TSV in “normal” seasons. • Survey New South Wales and Queensland crops and determine alternative weed and crop hosts. • Assess yield-loss in cotton due to TSV, and factors that lead to systemic infection. • Assess thrips vector species present in cotton • Provide extension material on the impact and management of TSV in cotton

重组竹叶青蛇毒纤溶酶原激活剂TSV-PA效应观察体内溶栓

】目的观察重组竹叶青蛇毒纤溶酶原激活剂一体内溶栓效应。方法利用家兔制备的脑血检 模型, 以生理盐水和尿激酶为对照, 用数字减影血管造影技术观察重组竹叶青蛇毒纤溶酶原激活剂飞体内溶 检效果。结果昭高剂量实验组喇再通时间为以内月州低剂量实验组止以内再 通。尿激酶组以刃再通时间为一以内生理盐水对照组没有观察到再通现象。实验动物后处 死解剖, 未观察到出血现象和其它异常现象。结论重组巧勺一在体内具有良好的溶检活性。

马氏珠母贝(Pinctada matensii)3个DM结构域的克隆及序列分析

Dmr(tdouble sex and Mab-3 related transcription factor)是新发现的与动物性别决定和分化相关的基因家族(Kondo et al.,2002),在多种进化地位的物种中都具有高度保守性。然而在作为动物界第二大门的贝类中,是否也存在高度保守的Dmrt基因家族,是否具有丰富的多样性?到目前为止未见任何相关报道。马氏珠母贝是我国人工培育海水珍珠的最主要母贝,在其养殖群体中有少数雌雄同体个体,并在一定条件下出现性转化。因此,克隆鉴定马氏珠母贝的Dmrt基因既可丰富D

中华鳖病毒(TSV)对鱼类细胞的感染试验

将 12种鱼类培养细胞用于中华鳖病毒 ( Trionyx sinesisvirus,TSV)的体外感染试验。结果显示 ,TSV能使鲤鱼上皮乳状瘤细胞 ( EPC)、鳗鲡性腺细胞 ( EG)和草鱼卵巢细胞 ( CO)产生病变。比较了不同培养温度对 TSV感染鱼类细胞的影响以及 TSV引起鱼类细胞病变的特征。 TSV引起 3种鱼类细胞病变 ,经显微观察 ,它们具有不同的病变特征。 TSV感染细胞后 ,其 TCID5 0 在 2 0～ 30℃范围内 ,随温度的升高而增加。超微观察表明 ,从感染的鱼类培养细胞中分

DM对大叶相思苗木生长的作用研究

研究了DM(DesertMagic,一种液体保水剂)与大叶相思苗木生长的关系.结果表明,使用DM能加速苗木的生长发育;苗高、地径、根长等生长指标明显高于对照;0～30cm土壤含水量显著提高.使用DM对培育大叶相思优质壮苗有良好的效果.

KDD, SEMMA and CRISP-DM: a parallel overview

In the last years there has been a huge growth and consolidation of the Data Mining field. Some efforts are being done that seek the establishment of standards in the area. Included on these efforts there can be enumerated SEMMA and CRISP-DM. Both grow as industrial standards and define a set of sequential steps that pretends to guide the implementation of data mining applications. The question of the existence of substantial differences between them and the traditional KDD process arose. In this paper, is pretended to establish a parallel between these and the KDD process as well as an understanding of the similarities between them.

Ciblage exosomal et effet de HLA-DM sur la présentation antigénique par le complexe majeur d'histocompatibilité de classe II

Les molécules du complexe majeur d'histocompatibilité de classe II (CMH II) sont exprimées exclusivement à la surface des cellules présentatrices d'antigènes et servent à stimuler les cellules CD4+ initiant une réponse immunitaire. Le chargement peptidique sur HLA-DR se produit dans les endosomes tardifs et les lysosomes sous l'action de HLA-DM. Cette molécule de classe II non-classique enlève les fragments peptidiques de la chaîne invariante (Ii) restés associés aux molécules de classe II (CLIP) et édite leur répertoire d'antigènes présentés. En utilisant une forme mutante de HLA-DM (HLA-DMy) qui s'accumule à la surface plasmique, nous avons observé que HLA-DMy augmente les chargements de peptides exogènes et aussi la réponse des cellules T en comparaison avec HLA-DM sauvage. Il a été démontré que des molécules chimiques, comme le n-propanol, pouvait avoir le même effet que HLA-DM en remplaçant les peptides associés aux molécules de classe II de la surface cellulaire. De plus, HLA-DMy et le n-propanol ont présenté un effet additif sur la présentation de peptides exogènes. Certaines protéines de la voie endocytique, comme HLA-DR, HLA-DM, HLA-DO et Ii sont ciblés aux compartiments multivésiculaires (MVB) et peuvent être ciblées aux exosomes. Suite à une fusion entre les MVB et la membrane plasmique, les exosomes sont relâchés dans le milieu extracellulaire. Nous avons déterminé que le motif tyrosine de HLA-DMβ et son interaction avec HLA-DR n'affectaient pas le ciblage aux exosomes, sauf la molécule HLA-DO. Cette étude nous a permis de démontrer que HLA-DMy augmente la quantité de peptides exogènes chargés sur les CPA et que HLA-DM et HLA-DMy sont incorporés dans les exosomes.

A importância dos contextos na transição para a vida pós-escolar do jovem com deficiência mental (dm): perspectivas dos técnicos, encarregados de educação e empresários

O presente trabalho apresenta uma análise aos diferentes factores que exercem influência, na fase de transição para a vida pós-escolar, dos jovens com Deficiência Mental. Esta análise incide na importância dos diversos contextos em que estes jovens estão inseridos (familiar, social, pessoal e escolar) e é aqui apresentada em três perspectivas diferentes: dos técnicos, dos encarregados de educação e dos empresários. Ao longo do trabalho são abordadas diferentes problemáticas que estes jovens experienciam na sua fase de transição da escola para o mundo do trabalho e expressas as opiniões marcadas dos diferentes grupos intervenientes no estudo. Estas opiniões constituem um conjunto identificado de problemas/soluções que acompanham a transição para a vida pós-escolar dos jovens portadores de Deficiência Mental.

Determination by MAD-DM of the structure of the DNA duplex d[ACGTACG(5-BrU)]2 at 1.46 Å and 100 K

A four-wavelength MAD experiment on a new brominated octanucleotide is reported here. d[ACGTACG(5-BrU)], C77H81BrN30O32P7, (DNA) = 2235, tetragonal, P43212 (No. 96), a = 43.597, c = 26.268 Å, V = 49927.5 Å3, Z = 8, T = 100 K, R = 10.91% for 4312 reflections between 15.0 and 1.46 Å resolution. The self-complementary brominated octanucleotide d[ACGTACG(5-BrU)]2 has been crystallized and data measured to 1.45 Å at both 293 K and a second crystal flash frozen at 100 K. The latter data collection was carried out to the same resolution at the four wavelengths 0.9344, 0.9216, 0.9208 and 0.9003 Å, around the Br K edge at 0.92 Å and the structure determined from a map derived from a MAD data analysis using pseudo-MIR methodology, as implemented in the program MLPHARE. This is one of the first successful MAD phasing experiments carried out at Sincrotrone Elettra in Trieste, Italy. The structure was refined using the data measured at 0.9003 Å, anisotropic temperature factors and the restrained least-squares refinement implemented in the program SHELX96, and the helical parameters are compared with those previously determined for the isomorphous d(ACGTACGT)2 analogue. The asymmetric unit consists of a single strand of octamer with 96 water molecules. No countercations were located. The A-DNA helix geometry obtained has been analysed using the CURVES program.