Interaction of a plasminogen activator proteinase, LV-PA with human alpha2-macroglobulin.

Lachesis venom plasminogen activator (LV-PA) is a 33-kDa serine proteinase isolated from bushmaster (Lachesis muta muta) snake venom, which activates the fibrinolytic system in vitro. This study has examined the effect of the plasma proteinase inhibitor alpha2-macroglobulin (alpha2-M) towards LV-PA and compares it with the effect on tissue type plasminogen activator (t-PA). The proteolytic activity of LV-PA alone or previously incubated with human plasminogen (Plg) on the large molecular mass protein substrates, dimethylcasein (DMC) and fibrinogen (Fg) was completely inhibited by human alpha2-M. However, the synthetic peptides Tos-Gly-Pro-Lys-pNA and H-D-Pro-Phe-Arg-pNA (S-2302) were hydrolyzed with almost no reduction in rate. At pH 7.4 and 37 degrees C the proteinase (0.15 microM over 15 min) interacted with alpha2-M, and each mole of alpha2-M bound 2 mol of enzyme. Sodium dodecyl sulfate gel electrophoresis of reduced samples showed that the interaction of alpha2-M with either LV-PA or t-PA preincubated with Plg resulted in the formation of approximately 90 kDa fragments and high molecular mass complexes (Mr 180 kDa), generated by the incubation mixture (LV-PA or t-PA) and Plg. The data suggest that LV-PA is a direct-type PA and its fibrinolytic effect can be reduced by alpha2-M in vivo.     

Ethical Reasoning Among Registered Nurses Experienced in Dementia Care

Abstract. Registered nurses regarded as “experienced and good” in dementia care were interviewed about the feeding of a severely demented patient who showed refusal-like feeding behaviour. Not one of the twenty nurses could see herself using force against her patients. Most interviewees justified their decisions to feed a severely demented patient and answered questions about whether they would change their minds if there were certain circumstances in terms of words that could be interpreted as referring to the ethical principle of beneficence. The nurses stressed the difficulty to understand the meaning of severely demented patients' feeding behaviour and decide when force-feeding occurs. When asked to rank ethical principles of importance for the decision, however, the most common answer was that they would give priority to the ethical principle of autonomy. The nurses did not see the ethical principles as separate entities, that could be applied one by one, but tried to integrate them into a whole. The findings of this study were interpreted as indicating that principled ethics is not an adequate model to describe experienced nurses' ethical reasoning.     

人胎儿雪旺氏细胞的体外培养及其纯化研究

本实验用酶消化法从孕１４～１９周人胎儿周围神经中分离培养雪旺氏细胞，结合差速贴壁和阿糖胞苷处理进行纯化，根据Ｓ－１００蛋白免疫细胞化学染色和形态学特征，分析不同时期雪旺氏细胞的纯化程度。实验结果显示：培养４ｄ，倒置相差显微镜下观察，雪旺氏细胞呈典型的双极或三极形，端对端或旋涡状排列，Ｓ－１００蛋白免疫细胞化学染色呈阳性反应，其比例约占９８％；在２～３周内，雪旺氏细胞的纯度维持在８５％～９０％；培养３５ｄ，雪旺氏细胞约占８０％；单纯采用阿糖胞苷或差速贴壁处理，培养３５ｄ，雪旺氏细胞约占７０％；原代培养９２ｄ，传代１１代细胞生长良好。实验结果表明，多种方法联合使用较单一方法所得的纯度高。本方法快速、简便，能为雪旺氏细胞作为移植材料提供足够的来源。     

中国西部城市和农村开展面粉营养素强化的可行性分析

目的通过对甘肃农产区居民的饮食习惯、营养知识和态度的调查,从需方的角度对开展面粉营养素强化的可行性进行探讨。方法采用定性和定量调查相结合的方法,对4个地区的城市和农村的男性和女性分别进行现状和原因的调查分析。结果被调查地区多数城市(92.7%)和农村(60.7%)家庭日常饮食以面食为主,94.7%的城市家庭购买面粉,而在农村地区,多数家庭自己种植或购买小麦。城市居民较农村对营养和维生素有更多的了解和关注。农村男性对面粉营养素强化持怀疑态度,女性更加关注孩子的营养,认为目前的饮食单一,孩子可能营养不足。结论与农村地区相比,城市地区的社会经济条件和知识基础较利于开展面粉强化工作。     

Induction of mosquitocidal activity in mice immunized with Anopheles gambiae midgut cDNA

Vaccines that induce mosquito-killing (mosquitocidal) activity could substantially reduce the transmission of certain mosquito-borne diseases, especially vaccines against African malaria vectors, such as the mosquito Anopheles gambiae. To generate and characterize antimosquito immunity we immunized groups of mice with two individual A. gambiae midgut cDNAs, Ag-Aper1 (a secreted peritrophic matrix protein) and AgMuc1 (a midgut-bound mucin), and an A. gambiae midgut cDNA library from blood-fed mosquitoes. We observed significantly increased mortality among mosquitoes that fed on either the AgMuc1- or the cDNA library-immunized mice compared to that of controls, but no differences were observed among those fed on Ag-Aper1-immunized mice. Analysis of the humoral and cellular immune responses from mice showed that the induced mosquitocidal effect was associated with immune profiles characterized by elevated tumor necrosis factor alpha and gamma interferon cytokine levels and very low antibody titers. Furthermore, an additional immunization of cDNA library-immunized mice with midgut protein shifted immunity toward a Th2-type immune response, characterized by elevated antibody titers and high interleukin-5 and interleukin-10 cytokine levels; importantly, mosquitoes feeding on these mice exhibited no undue mortality. Finally, when immune sera was ingested by mosquitoes through a membrane feeder, no effect on mosquito mortality was observed, indicating that serum factors alone were not responsible for the mosquitocidal effect. Our results demonstrate that mosquitocidal immunity in mice can be consistently generated by midgut cDNA immunization and suggest this cDNA-induced mosquitocidal immunity is cell mediated.