ExoU, uma proteína do sistema de secreção do tipo III de Pseudomonas aeruginosa, induz a secreção de eicosanóides por células epiteliais respiratórias. Papel dos corpúsculos lipídicos neste processo

Para pesquisar o papel de ExoU no desencadeamento de resposta inflamatória nas vias aéreas, células epiteliais respiratórias humanas (CERs) da linhagem BEAS-2B foram tratadas com AA radiomarcado e infectadas com a cepa PA103 de P. aeruginosa, que secreta ExoU, e com os mutantes PA103exoU (com deleção do gene exoU), PA103ΔUT/exoU (com deleção de exoU e complementação com o gene funcional) e PA103UT/S142A (com deleção de exoU e complementação com gene com mutagênese sítio-específica no domínio catalítico da enzima). Após 1 hora, a liberação de AA pelas culturas infectadas com as cepas produtoras de ExoU foi significativamente superior à observada em culturas infectadas pelas cepas não-produtoras ou por células controle. O tratamento das bactérias com MAFP, um inibidor de PLA2, resultou em significativa redução na liberação de AA. Células infectadas pelas cepas PA103 e PA103ΔUT/exoU secretaram PGE2 e LTB4 em concentrações significativamente maiores que as secretadas por células infectadas pelas demais cepas ou não infectadas. O tratamento com o MAFP reduziu significativamente a secreção de PGE2. A análise, por citometria de fluxo, de células infectadas e não infectadas tratadas com anticorpo anti-COX-2 mostrou que o percentual de células infectadas por PA103 marcadas foi significativamente superior ao percentual encontrado em culturas controle. Nenhuma diferença foi observada quanto ao percentual de células marcadas em culturas infectadas por PA103ΔexoU. O tratamento das culturas com NS-398 (um inibidor seletivo de COX-2) resultou na diminuição significativa da concentração de PGE2, secretada por células infectadas com PA103, mas não por células infectadas com PA103ΔexoU ou por células controle. Corpúsculos lipídicos (CLs) são domínios citoplasmáticos ricos em COX-2 e outras enzimas responsáveis pelo metabolismo do AA, sede da produção de eicosanóides. Como células infectadas pelas cepas produtoras de ExoU liberam AA livre, formulamos a hipótese de que a maior produção de eicosanóides por estas células seria dependente da indução do aumento no número dos CLs. No entanto, a análise por citometria de fluxo de células tratadas com uma sonda lipofílica com afinidade com os CLs mostrou que o percentual de células marcadas em culturas infectadas pelas cepas produtoras de ExoU foi significativamente inferior ao percentual em culturas controle ou infectadas pelas outras 2 cepas bacterianas. O tratamento das células com MAFP inibiu significativamente a redução do percentual de células contendo CLs. A análise, por citometria de fluxo, de células controle ou infectadas tratadas simultaneamente com a sonda lipofílica e com o anticorpo anti-PGE2, mostrou, em células infectadas com PA103, a redução da mediana da intensidade de marcação com a sonda lipofílica e o aumento da mediana da intensidade de marcação com o anticorpo anti-PGE2. Nossa hipótese é que a presença de ExoU nas células infectadas com a cepa PA103 resulte no metabolismo de glicerofosfolipídios presente nos CLs levando à diminuição da afinidade dos CLs pela sonda lipofílica e à síntese local de PGE2.

Reproductive biology of Black Anglerfish (Lophius budegassa) in the northwestern Mediterranean Sea

Gonadal morphology and reproductive biology of the Black Anglerfish (Lophius budegassa) were studied by examining 4410 specimens collected between June 2007 and December 2010 in the northwestern Mediterranean Sea. Ovaries and testes presented traits common among fishes of the order Lophiiformes. Spawning occurred between November and March. Size at first maturity (L50) was 33.4 cm in total length (TL) for males and 48.2 cm TL for females. Black Anglerfish is a total spawner with group-synchronous oocyte development and determinate fecundity. Fecundity values ranged from 87,569 to 398,986 oocytes, and mean potential fecundity was estimated at 78,929 (standard error of the mean [SE] 13,648) oocytes per kilogram of mature female. This study provides the first description of the presence of 2–3 eggs sharing the same chamber and a semicystic type of spermatogenesis for Black Anglerfish. This new information allows for a better understanding of Black Anglerfish reproduction—knowledge that will be useful for the assessment and management of this species.

中国产三种野生稻比较胚胎学研究

普通野生稻是热带、亚热带分布种。在我国主要分布于南方几个省区。普通野生稻与栽培稻同属AA型染色体组，与栽培稻亲缘关系最近，可能是栽培稻的祖先种。从农学和育种学角度对普通野生稻开展了不少研究工作。但对其胚胎学方面的工作，似乎无人涉及。本文针对这一情况而展开工作，主要结果如下： 1．花药还处在造孢时期，花药壁层由4-5层细胞组成，而且被胞间连丝联结，其造孢细胞间存在胞间连丝和胞质通道． 2．小孢子母细胞减数分裂时，其胞质分裂是连续型，小孢子呈左右对称排列。在小孢子和花粉粒发育过程中，有明显极性存在。在小孢予液泡化后期，小孢子核和细胞质远离小孢子萌发孔和绒毡层呈极性分布o在二核花粉粒时期，花粉粒中淀粉粒的积累也呈极性方式，即从萌发孔处开始积累，向远离萌发孔处推进。 3．花粉粒中的二精子，具有“头尾一状结构。精细胞与不规则形状的营养核存在空间上的物理联结。 4．在自由小孢子期，绒毡层内质网发生叠堆，可能是对小孢子液泡化后产生的膨压和张力的一种适应。绒毡层在小孢子液泡化中期开始退化，开始退化的绒毡层表现为内质网腔膨大，核周腔增大，核孔数增加． 5．在二胞花粉时期，内壁形成之前，出现4-5层的层状结构，这在禾本科其它植物未曾观察到。 6．普通野生稻花药开裂与双子叶植物明显不同，没有circular cell clus-ter和stomium结构。其花药开裂通过“开裂腔一来完成的，败育的花药不形成开裂腔，并建立了模型。 7．胚囊发育为蓼型，反足细胞分裂形成反足组织，反足组织含有大量淀粉粒，反足细胞直到胚乳细胞化时才退化，反足细胞在胚和胚乳发育中起着积极作用． 8．合子在受精后约6小时开始横向分裂，形成两个大小不等原胚细胞，接着顶细胞第二次纵向分裂。在原胚期，其基细胞具淀粉粒，且细胞通过胞间连丝相互联系．胚柄伸入珠心组织直至子房壁，为营养物的输入提供了有效的通道． 9．胚乳发育属核型。胚乳细胞化通过初期自由生长壁形成细胞壁，后期通过成膜体和细胞板的方式共同形成细胞壁。刚形成的胚乳细胞通过胞间连丝相连，并含有各种细胞器。 10.通过对8个居群(群体)398个子房切片观察，在普通野生稻中未发现有无融合生殖现象。

Impact of mechanisation of fisheries development in Karnataka

Karnataka, one of the maritime states on the west coast of India, has progressed quite well in marine fisheries due to its vast fisheries resources and diversification in mechanised fishing. Mechanisation programmes were started by the state from 1957-58 only. Starting with two small mechanised boats, the state has today a fishing fleet of 398 purseseiners, 731 gillnetters, 2 deepsea trawlers and about 1,500 shrimp trawlers contributing over 85% of the total marine fish landings. The marine fish production during 1987-88 up to the end of March 1988 was 1,29,659 tonnes valued at Rs.48.05 crores.

Phylogenomic study of the subfamily Caprinae by cross-species chromosome painting with Chinese muntjac paints

Chromosomal homologies have been established between the Chinese muntjac (Muntiacus reevesi, MRE, 2n = 46) and five ovine species: wild goat (Capra aegagrus, CAE, 2n = 60), argall (Ovis ammon, OAM, 2n = 56), snow sheep (Ovis nivicola, ONI, 2n = 52), red goral (Naemorhedus cranbrooki, NCR, 2n = 56) and Sumatra serow (Capricornis sumatraensis, CSU, 2n = 48) by chromosome painting with a set of chromosome-specific probes of the Chinese muntjac. In total, twenty-two Chinese muntjac autosomal painting probes detected thirty-five homologous segments in the genome of each species. The chromosome X probe hybridized to the whole X chromosomes of all ovine species while the chromosome Y probe gave no signal. Our results demonstrate that almost all homologous segments defined by comparative painting show a high degree of conservation in G-banding patterns and that each speciation event is accompanied by specific chromosomal rearrangements. The combined analysis of our results and previous cytogenetic and molecular systematic results enables us to map the chromosomal rearrangements onto a phylogenetic tree, thus providing new insights into the karyotypic evolution of these species.

非晶状体βγ-晶状体蛋白与三叶因子蛋白复合物的细胞核转运及其选择性杀伤肿瘤细胞机制的研究

非晶状体βγ晶状体蛋白与三叶因子蛋白复合物(βγ-CAT)是从大蹼铃蟾(Bombina maxima)皮肤分泌物中分离的分子量为72 kDa的天然蛋白复合物.本研究通过激光共聚焦显微镜和Westem blot分析βγ-CAT在人脐静脉内皮细胞(HUVEC)中的细胞核转运机制,以及βγ-CAT对多株肿瘤细胞(HCT116,HT29,A375,Hela,THP-1等)的细胞毒效应.结果表明:βγ-CAT的α亚基中含有典型的GTP/ATPase的保守结构模体Walker A和Walker B,体外检测到βγ-CAT具有GTP/ATP水解酶和GTP/ATP结合活性.在细胞核转运过程中,βγ-CAT的α亚基和β亚基参与形成约150kDa含有泛素化修饰信号的大分子复合物,且泛素化修饰信号和βγ-CAT的α亚基和β亚基共定位于细胞内和融合于细胞核区域的转运囊泡小体中.βγ-CAT能够选择性的杀伤肿瘤细胞,诱导肿瘤细胞脱落和发生凋亡.上述结果为进一步深入研究阡βγ-CAT的细胞核转运和调节细胞功能的分子作用机制提供思路和线索.

A phylogeny of Chinese species in the genus Phrynocephalus (Agamidae) inferred from mitochondrial DNA sequences

We investigated the phylogenetic relationships among most Chinese species of lizards in the genus Phrynocephalus (118 individuals, collected from 56 populations of 14 well-defined species and several unidentified specimens) using four mitochondrial gene fragments (12S rRNA, 16S rRNA, cytochrome b, and ND4-tRNA(LEU)). The partition-homogeneity tests indicated that the combined dataset was homogeneous, and maximum-parsimony (MP), neighbor-joining (NJ), maximum-likelihood (ML) and Bayesian (BI) analyses were performed on this combined dataset (49 haplotypes including outgroups for 2058 bp in total). The maximum-parsimony analysis resulted in 24 equally parsimonious trees, and their strict consensus tree shows that there are two major clades representing the Chinese Phrynocephalus species: the viviparous group (Clade A) and the oviparous group (Clade B). The trees derived from Bayesian, ML. and NJ analyses were topologically identical to the MP analysis except for the position of P. mystaceus. All analyses left the nodes for the oviparous group, the most basal clade within the oviparous group, and P. mystaceus unresolved. The phylogenies further suggest that the monophyly of the viviparous species may have resulted from vicariance, while recent dispersal may have been important in generating the pattern of variation among the oviparous species. (C) 2003 Elsevier Science (USA). All rights reserved.