38 resultados para isogenic

em Repositório Institucional UNESP - Universidade Estadual Paulista "Julio de Mesquita Filho"


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Toxoplasmosis is an anthropozoonotic widespread disease, caused by the coccidian protozoan parasite Toxoplasma gondii. Since there are no data regarding the genotoxicity of the parasite in vivo, this study was designed to evaluate the genotoxic potential of the toxoplasmosis on isogenic mice with normal diet or under dietary restriction and submitted to a treatment with sulfonamide (375 mug/kg per day). DNA damage was assessed in peripheral blood, liver and brain cells using the comet assay (tail moment). The results for leucocytes showed increases in the mean tail moment in mice under dietary restriction; in infected mice under normal diet; in infected, sulfonamide-treated mice under normal diet; in infected mice under dietary restriction and in infected sulfonamide-treated mice under dietary restriction. In liver and brain cells, no statistically significant difference was observed for the tail moment. These results indicated that dietary restriction and T. gondii were able to induce DNA damage in peripheral blood cells, as detected by the comet assay. (C) 2004 Elsevier B.V. All rights reserved.

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In order to study the morphological changes that occur in cells of the testes of isogenic black mouse C57BL/6/Uni into three periods during spermatogenetic used 15 mice divided into 3 groups of 5 animals with 40,50 and 60 days of age. The mice were sacrificed and weighed. Testicles were weighed and measured, and histologically processed and stained with HE, PAS and Masson Massom-H and evaluated under light microscopy. It was observed that group I with 40 days of age in the seminifcrous tubules had a lumen with sparse small amount of interstitial tubular cells. In the seminiferous epithelium type A spermatogonia, intermediate and B were identified, which occupied the compartment adbasal and intermingled with these cells in spermatocytes I in Pachytene and leptotene was observed, whereas in the adluminal compartment Golgi phase spermatids we observed the presence of acrosomal granule. In group II, the cells of the seminiferous epithelium were developed and it was observed in round spermatids cephalic hood phase plus many elongated spermatids in acrosome phase and Sertoli cells. In Group III, 60 days old, it was found that seminiferous epithelium which was of the tubules had elongated spermatids in acrosome phase and maturation, with elongated nuclei and acrosomal system typical of spermiation in the presence of sperm and residual bodies near the tubular lumen. Therefore morphological evolution of germ cell testicular spermatids can be checked and recognized in its four phases: Golgi, cap, acrosome and maturation over the age of the animal.

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Morphological investigations of the epithelial cell types that line the ductuli efferentes (DE) of black isogenic mice confirm absorption of the luminal fluid phase by endocytosis as the main function of ductuli efferentes (DE) in this species. :Furthermore, all the histochemical and ultrastructural observations on the DE epithelial histoarchitecture indicate other cellular functions such as exocytosis and probably secretion: including an aprocrine secretory process.

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Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

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Morphological and functional alterations caused by chronic alcohol ingestion in testes and accessory sex organs have been studied both in man and in laboratory animals. The aim of the present study was to examine the possible occurrence of deleterious effects of chronic alcohol ingestion on the secretory epithelium of the ventral prostate of mice. Twenty-four adult male C57BL/6J mice were divided into three groups. The alcohol-treated group was allowed to drink only 6% (v/v) ethanol, the isocaloric group received a diet of water/sucrose with a calorie content equivalent to a 6% alcohol solution and the control group received water. Both groups were fed ad libitum with solid Purina rat chow. After 120 days, animals from each group were anesthetized with ethyl ether, weighed and processed for light and transmission electron microscopy. The results demonstrated reduction in the glandular epithelium cell height and disorganization of the Golgi complex. Moreover, abundant membrane-bound structures, most likely representing cytoplasmic material, were observed, as well as accumulation of dense bodies. Statistical analysis showed that bodyweight gain was similar for both groups. In conclusion, chronic alcohol ingestion has harmful effects on the secretory epithelium cells of the ventral lobe of the prostate of mice after 120 days of treatment. (C) 2001 Harcourt Publishers Ltd.

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The specific delayed-type hypersensitivity (DTH) response was evaluated in resistant (A/SN) and susceptible (B10.A) mice intraperitoneally infected with yeasts from a virulent (Pb18) or from a non-virulent (Pb265) Paracoccidioides brasiliensis isolates. Both strains of mice were footpad challenged with homologous antigens. Pb18 infected A/SN mice developed an evident and persistent DTH response late in the course of the disease (90th day on) whereas B10.A animals mounted a discrete and ephemeral DTH response at the 14th day post-infection. A/SN mice infected with Pb265 developed cellular immune responses whereas B10.A mice were almost always anergic. Histological analysis of the footpads of infected mice at 48 hours after challenge showed a mixed infiltrate consisting of predominantly mononuclear cells. Previous infection of resistant and susceptible mice with Pb18 did not alter their DTH responses against heterologous unrelated antigens (sheep red blood cells and dinitrofluorobenzene) indicating that the observed cellular anergy was antigen-specific. When fungal related antigens (candidin and histoplasmin) were tested in resistant mice, absence of cross-reactivity was noted. Thus, specific DTH responses against P. brasiliensis depend on both the host's genetically determined resistance and the virulence of the fungal isolate.

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Durante o armazenamento de grãos de feijão (Phaseolus vulgaris L.), o rendimento pode ser reduzido devido às infestações de carunchos como os da espécie Zabrotes subfasciatus (Bohemann, 1833) (Coleoptera: Bruchidae). O ataque desse inseto afeta diretamente a qualidade dos grãos, além de facilitar a entrada de patógenos, tornando-os inviáveis para o consumo e/o comércio. Com a finalidade de buscar uma estratégia alternativa para o controle deste caruncho, avaliou-se a possível resistência de linhagens quase isogênicas contendo arcelina, linhagens selvagens contendo arcelina e cultivares comerciais de feijoeiro, em laboratório (T= 25±2° C, U.R.= 70±10% e fotoperíodo= 12h). Foram utilizados frascos contendo 10 g de grãos dos genótipos, os quais foram infestados por uma semana com sete casais do caruncho. Vinte e um dias após a infestação, os grãos foram avaliados contando-se o número de ovos viáveis. A partir de 25 dias da infestação, os grãos foram observados diariamente avaliando-se o número e o peso dos insetos emergidos, a viabilidade larval, o ciclo biológico (ovo-adulto) e o peso de grãos consumidos. Empregou-se um delineamento inteiramente casualizado, com oito repetições. Os genótipos Arc.2, Arc.3, Arc.4, Arc.3S e Ipa 6 expressaram baixos níveis de não-preferência para oviposição e foram classificados como deterrentes. Os genótipos Arc.1S e Arc.1 expressaram elevados níveis de antibiose; Arc.2, Arc.3 e Arc.4 apresentam o mesmo mecanismo, porém, em níveis inferiores.

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Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

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The mechanisms by which arthritis-provoking pathogens such as Yersinia enterocolitica interact with the human immune system to produce inflammatory synovitis are not well known. One of the immunomodulating mechanisms used against these pathogens is the polyclonal activation of lymphocytes. In this study, we investigated the extent of the B-lymphocyte activation induced in mice by a strain of Y. enterocolitica O:3 (FCF 526) isolated from a patient with arthritis, and compared it with two other strains, a virulent one (FCF 397[+]) isolated from a patient without arthritis and its plasmidless isogenic pair (FCF397[-]). Also we investigated the production of autoantibodies in mice infected with these different strains. SPF Swiss mice were infected intravenously with a suspension of Y. enterocolitica . Spleen cells were taken on days 7, 14, 21 and 28 after infection and the number of cells secreting nonspecific and specific antibodies of IgG 1 , IgG 2a , IgG 2b , IgG 3 , IgM and IgA isotypes were determined by the ELISPOT technique. The presence of autoantibodies in mouse serum was investigated by the dot-blot assay. The pattern of infection of the three bacterial strains were almost the same. We observed a general increase in the number of nonspecific Ig-secreting cells with all three strains, and the greatest increases observed were in the IgG 2a and IgG 3 isotypes. Only a small fraction of the immunoglobulins detected were antibacterial, suggesting that the rest resulted from polyclonal B cell activation. The strain isolated from the patient with arthritis (FCF526) induced the greatest production of autoantibodies, coinciding with the period in which the greatest activation of nonspecific B lymphocytes was seen. There were no signs of arthritis or inflammation in the joints of the infected animals. Based on our results, we were unable to determine whether there is an association between the arthritogenic capability of Y. enterocolitica and polyclonal activation of B cells.

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The Brazilian Agency for the Environment (IBAMA) recently adopted an alternative medium-term multiple-organ assay system with the Wistar rat strain for detection of the carcinogenic potential of pesticides. Originally, this initiation-promotion protocol was established in Japan with the isogenic Fischer 344 male rat. Among the initiating agents used in that assay, N-methyl-N-nitrosourea (MNU) rapidly induces malignant lymphoma and leukemia and early mortality of rats from different strains. This study was developed to evaluate whether the outbred Wistar rats are also similarly susceptible to MNU. Particularly, it aimed to evaluate the dose-response relationship and to register the MNUinduced pre-neoplasia and neoplasia that may develop in the lympho-hematopoietic system (LHS) of the Wistar rat within a medium-term period. Four groups of male Wistar rats were treated during 2 weeks with vehicle or with MNU (80, 160 or 240 mg/kg body weight, i.p.). After sacrifice at the 12th and 20th weeks, the thymus, spleen, bone marrow, cervical and mesenteric lymph nodes and liver were collected for analysis. At the 20th week, LHS malignant tumors and benign vascular tumors occurred only in the high- and intermediate-dose MNU-treated animals. Four animals treated with 240 mg/kg developed diffuse thymic lymphomas; two others, treated respectively with 240 mg/kg and 160 mg/kg, developed spleen hemangiomas. The present observations indicate that the Wistar strain is as susceptible as other strains to the early development of MNU-induced LHS (pre)neoplasia. Therefore, this strain seems suitable to be used as test system in bioassay protocols that adopt MNU as an initiating agent for carcinogenesis.

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Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

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Embora não haja cultivos comerciais de milho geneticamente modificado no Brasil, o efeito de híbridos de milho Bt sobre inimigos naturais e artrópodos de solo deve ser avaliado antes da liberação aos produtores. Assim, ensaios foram conduzidos durante uma safra em duas localidades. Os híbridos de milho modificado geneticamente 7590-Bt11 e Avant-ICP4 foram comparados com seus respectivos isogênicos não transgênicos. Os artrópodes foram avaliados através de observação direta nas plantas e armadilhas de alçapão. de modo geral, não se observaram diferenças entre as populações de tesourinha (Dermaptera: Forficulidae), joaninhas (Coleptera: Coccinellidae), percevejo-pirata (Coleoptera: Anthocoridae), carabídeos (Carabidae), cicindelídeos (Cicindelidae) e aranhas (Araneae). Também não houve diferença no parasitismo de ovos de Helicoverpa zea (Boddie) por Trichogramma sp. (Hymenoptera: Trichogrammatidae). Assim, milho geneticamente modificado expressando as proteínas inseticidas Cry1A(b) e VIP 3A não causa redução nas populações dos principais predadores e parasitóides.

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Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

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A biologia de Bemisia tabaci biótipo B (Genn.) foi avaliada em genótipos de feijoeiro (Phaseolus vulgaris L.) que contêm arcelina em suas sementes. Foi também realizada análise bioquímica de proteínas, em sementes e em folhas dos genótipos de feijoeiro, a fim de verificar se haveria traços de arcelina nas folhas dos materiais a serem avaliados. Os testes foram conduzidos em condições de casa de vegetação, nas épocas das águas e da seca, em dois anos consecutivos, com os seguintes genótipos: ARC 3s, ARC 5s (genótipos selvagens portadores de arcelina); ARC 1, ARC 2, ARC 3, ARC 4 (linhagens quase-isogênicas portadoras de arcelina - Empresa Brasileira de Pesquisa Agropecuária (EMBRAPA)), Porrillo 70, Bolinha e IAPAR MD 808 (genótipos sem arcelina). Os genótipos selvagens, ARC 3s e ARC 5s, apresentaram altos níveis de antibiose, com ênfase para o ARC 5s (as ninfas tiveram alta mortalidade, em torno de 90%). O prolongamento do ciclo de desenvolvimento dos insetos provenientes do genótipo ARC 5s podem sugerir uma resistência do tipo antibiose e/ou não-preferência para alimentação. A resistência dos genótipos selvagens não está relacionada com a presença de arcelina nas sementes, já que nenhum traço dessa proteína foi encontrado nas folhas destes.