Underlying mechanisms of schistosome/snail compatibility

说明(由申请人提供)：本建议书的目的包括 各种生化和分子方法来确定 抗性(R)与敏感(S)B的品系特异性差异。 这将导致对国防活动的更好理解 蜗牛。PI假设：(A)可能存在特定菌株的差异 在血细胞的呼吸爆发中；(B)存在敏感性差异 在S和R血细胞中对一种正常中间体的负性影响 呼吸爆发(如氯胺)和；(C)可能有菌株 诱导特定基因转录本的差异。 这项提案中的大部分工作都集中在体外试验的使用上。 用于确定孢子囊杀灭的系统，PI已经开发出 选择性地抑制O2和N依赖途径中的某些酶。 自上次续签以来，PI列出了11份手稿(出版和在 新闻/提交)和8篇评论文章涉及这一问题的各个方面 蜗牛/吸虫的关系。其他几篇没有直接关系的论文是 列出了作为展示PI在技术方面的专业知识的支持 以备将来研究之用。上一个资助期的调查结果导致了 了解到，至少在体外，氧气依赖和非依赖 机制可能在孢子囊的杀灭中起作用。这发生在中央军委之后。 化验有了很大的改进，发现过氧化氢参与了杀死 (尽管他们未能显示出R和S在ROS产生方面的差异 非CMC分析中的血细胞)。这些实验得出了这样一个前提： 抗性钉螺中孢子囊的破坏可能是由于有缺陷的 髓过氧化物酶(MPO)允许过氧化氢介导杀伤，而MO血细胞 使过氧化氢解毒，才能充分发挥其作用。 PI还发现了大量证据表明NO和过氧化氢在 孢子囊杀灭中的协同作用。这些结果将作为以下工作的基础 进一步探索这两条杀死孢子囊的途径。

DESCRIPTION (provided by the applicant): The aims of this proposal cover a variety of biochemical and molecular approaches to determine the strain-specific differences between resistant (R) vs. susceptible (S) B. glabratathat would lead to a greater understanding of defense activity in the snail. The PI hypothesizes that: (a) there can be strain-specific differences in the respiratory burst of hemocytes; (b) that sensitivity differences exist in the S and R hemocytes to negative influence of a normal intermediate of the respiratory burst (e.g. chlorinated amines) and; (c) there may be strain differences in induction of specific gene transcripts. Much of the work in this proposal centers on the use of an in vitro assay system for determining sporocyst killing, and the PI has developed ways for selectively inhibiting certain enzymes in O2 and N dependent pathways. Since the last renewal the PI lists 11 manuscripts (published and in press/submitted) and 8 review articles dealing with various aspects of this snail/trematode relationship. Several other papers not directly related are listed that serve as support for demonstrating the PI's expertise in techniques for future studies. Findings during the last funding period led to the understanding that, at least in vitro, both O2 - dependent and -independent mechanisms may play a role in sporocyst killing. This came about after the CMC assay was substantially improved, and finding that H2O2 was involved in killing (although they failed to show differences in ROS production in R and S hemocytes in non-CMC assays). These experiments led to the premise that destruction of sporocysts in resistant snails may be due to a defective myeloperoxidase (MPO) that allows H2O2 to mediate killing, whereas MO hemocytes detoxify H2O2 before it is able to exert its effect fully. The PI also has developed substantial evidence that NO and H2O2 play synergistic roles in sporocyst killing. These results will serve as a basis to explore further these two pathways in sporocyst killing.