Spermatogenesis is a complex process of male germ cell development from diploid spermatogonia to haploid fertile spermatozoa. Apoptosis plays a vital role in limiting cell numbers and eliminating defective germ cells. This requires novel gene products, and precise and well-coordinated programmes of gene expression. It is therefore possible that a disruption of transcription factor function would significantly impact germ cell development. The present work was undertaken to use Staput separation followed by culture of purified germ cells of rodent testis since mammalian spermatogenesis cannot yet be recreated in vitro. Specificity of separation was assessed using immunocytochemistry to identify spermatogonia, spermatocytes and spermatids. The genotoxins H2O2, doxorubicin, N-ethyl-N-nitrosourea, N-methyl-N-nitrosourea, 6-mercaptopurine, 5-bromodeoxyuridine, methyl methanesulphonate and ethyl methanesulphonate were investigated. Cells were cultured and treated with different concentrations for each agent. DNA damage and apoptosis were measured by Comet and TUNEL assay respectively. Up-regulation of expression of the transcription factors Tbpl1, FHL5 and Gtf2a1l that are important post-meiotically, were examined using RT- PCR and qPCR. Protein production was evaluated using Western blotting. Tbpl1, FHL5 and Gtf2a1l were cloned in-frame into the inducible expression vector pET/100-TOPO. The recombinant clones were induced and successful expression of the proteins in E. coli was confirmed by SDS-PAGE and Western blotting. The recombinant clones obtained were used to demonstrate genotoxin induced impairment of gene expression. Thus, Staput-isolated rodent testicular germ cells seem to be a suitable model to study genotoxicity in vitro yielding result comparable to those reported in vivo. Furthermore, the work shows that genotoxins can impair gene expression.

A avaliação da mecânica respiratória (AMR) busca descrever o comportamento mecânico do sistema respiratório, para melhor entender a sua fisiologia e a sua patofisiologia. O método quasi-estático é caracterizado pela AMR em condições nas quais a aceleração dos tecidos pode ser considerada desprezível. Esta dissertação de mestrado teve como objetivo avaliar a mecânica respiratória pelo método quasi-estático em roedores, utilizando um insuflador pulmonar para pequenos roedores (IPPR) construído no Laboratório de Engenharia Biomédica (LEB) da Escola Politécnica da Universidade de São Paulo (EPUSP). Uma extensa revisão da literatura sobre fisiologia e técnicas de AMR em humanos e roedores é apresentada. Uma documentação bastante detalhada sobre a implementação e o funcionamento do IPPR é fornecida, com o intuito de permitir que o dispositivo seja reproduzido. Testes de bancada foram executados, para avaliar o funcionamento do IPPR, e um protocolo experimental com modelo animal de doença, em ratos Wistar envenenados por Paraquat, foi realizado. Os resultados dos testes de bancada mostraram que o equipamento ainda carece de alguns ajustes, pois apresenta erros na indicação de volume, da ordem de 5%, mas que podem ser compensados na análise dos dados experimentais obtidos. No futuro, pretende-se sanar essa imprecisão, tornando o IPPR independente de um microcomputador com sistema operacional multitarefa. Os resultados da aplicação do modelo matemático exponencial, nos dados obtidos de curvas pressão-volume no protocolo experimental, trouxeram informações que estão de acordo com as características observadas no modelo animal de envenenamento por Paraquat. / The assessment of respiratory mechanics (AMR) aims to describe the mechanical behavior of the respiratory system in order to better understand its physiology and pathophysiology. The quasi-static method is characterized by the AMR under conditions where the tissue\'s acceleration can be considered negligible. The purpose of this master degree dissertation was to assess the respiratory mechanics by quasistatic method in rats using a lung insufflator for small rodents (IPPR) implemented in the Biomedical Engineering Laboratory (LEB) of Escola Politécnica of University of São Paulo (EPUSP). Abroad literature review on AMR techniques and on physiology in humans and rodents was presented. A fairly detailed documentation of the assembly and the operation of the IPPR was provided in order to allow the device to be reproduced. Bench tests were conducted to evaluate the behavior of the IPPR, and an animal disease model by Paraquat poisoning protocol was performed in Wistar rats. The bench tests results showed that the equipment still needs some tweaking, because it presents errors in volume indication of the order of 5%, but these errors can be compensated in the analysis of the experimental data. In the future, there is an intention to correct this inaccuracy turning the IPPR independent of a multitask microcomputer operating system. The results of the mathematical exponential modeling applied on the pressure-volume curves in the experimental protocol are in good agreement with the characteristics observed on Paraquat poisoning animal model.

Made available in DSpace on 2016-04-28T20:15:35Z (GMT). No. of bitstreams: 1 2009 - Raquel de Oliveira Simoes.pdf: 2148721 bytes, checksum: 7408aa4a20471b27145d83c2bea4886a (MD5) Previous issue date: 2009-01-28 / Coordena??o de Aperfei?oamento de Pessoal de N?vel Superior / The objective of this study was to describe the composition and structure of helminthes community comparing the helminthes community component and infracommunity from sympatric rodents Akodon spp. and Oligoryzomys nigripes in the Atlantic Forest and in the different fragments size and the Parque Nacional da Serra dos ?rg?os (PARNA SO reserve. One hundred and fifty-six specimens of Akodon spp. and fifty-seven of Oligoryzomys nigripes were collected in Teres?polis, being 38 rodents in 4 small fragments (30 Akodon spp. and 8 O. nigripes), 58 in 4 medium fragments (43 Akodon spp. and 15 O. nigripes), 87 in 4 large fragments (67 Akodon spp. and 20 O. nigripes) and 30 in PARNASO (16 Akodon spp. e 14 O. nigripes). The stydy was done for two years (2004- 2005). The animals were necropsied and the viscera and cavities examined to helminthes parasites. A total of eighteen helminthes species were collected: 15 from Akodon spp. and12 from O. nigripes. Nine species were common for both rodents. The rodent Akodon spp. showed a higher richness than O. nigripes. A low degree of similarity between the helminthes infracommunities from Akodon spp. and O. nigripes was detected, although these rodents species live in coexistence. The terrestrial and arboreal behavior of O. nigripes and terrestrial behavior of Akodon spp. would determine this difference between both parasite communities. Probably, with the share of soil resource boths species of rodents could disseminate or be infected by the parasite. The prevalence of Stilestrongylus aculeata and mean abundance for Stilestrongylus eta increased, respectively, in small, medium and large fragments in Akodon spp.. The rodent O. nigripes showed high prevalence rate and mean abundance values for Stilestongylus lanfrediae (over 86%) regardless of the fragments size studied. The difference in mean richness for Akodon spp. and O. nigripes in different fragments size and PARNASO were not significant. There were variations between the helminthes infracommunities for the two species of rodent in different fragments size and PARNASO. These differences may also be related to the environmental changes that can produce both positive and negative effects on the transmission of helminthes parasites. / O objetivo deste trabalho foi descrever a composi??o e estrutura da comunidade helm?ntica comparando a comunidade componente e a infracomunidade dos helmintos das popula??es de roedores simp?tricos Akodon spp. e Oligoryzomys nigripes na Mata Atl?ntica e os diferentes tamanhos de fragmentos e a reserva do Parque Nacional da Serra dos ?rg?os (PARNASO). Cento e cinq?enta e seis esp?cimes de Akodon spp. e cinq?enta e sete de O. nigripes foram coletados no munic?pio de Teres?polis, sendo 38 roedores em 4 fragmentos pequenos (30 Akodon spp. e 8 O. nigripes), 58 em 4 fragmentos m?dios (43 Akodon sp p. e 15 O. nigripes), 87 em 4 fragmentos grandes (67 Akodon spp. e 20 O. nigripes) e 30 no PARNASO (16 Akodon spp. e 14 O. nigripes). O estudo foi feito durante dois anos (2004- 2005). Os animais foram necropsiados tendo suas v?sceras e cavidades tor?cica e abdominal examinadas para recupera??o de helmintos parasitos. Um total de 18 esp?cies de helmintos foram coletados: 15 em Akodon spp. e 12 em O. nigripes. Nove esp?cies foram comuns aos dois roedores. O roedor Akodon spp. teve uma maior riqueza parasit?ria do que O. nigripes. Um baixo grau de similaridade entre as infracomunidades de helmintos de Akodon spp. e O. nigripes foi observado, embora sejam esp?cies que coexistam. O comportamento terrestre e arbor?cola de O. nigripes e apenas terrestre de Akodon spp. poderia determinar essa diferen?a entre as duas comunidades de helmintos parasitos. Provavelmente, ao compartilharem os recursos do solo, ambas as esp?cies podem se infectar ou disseminar o parasita. A preval?ncia para Stilestrongylus aculeata e a abund?ncia m?dia para S. eta aumentaram, respectivamente, nos fragmentos pequenos, m?dios e grandes no Akodon spp.. O roedor O. nigripes apresentou elevada taxa de preval?ncia e valores de abund?ncia m?dia para S. lanfrediae (acima de 86%) independente do tamanho dos fragmentos estudados. A diferen?a na riqueza m?dia de Akodon spp. e O. nigripes nos diferentes tamanhos de fragmentos e no PARNASO n?o foi significativa. Houve varia??es entre as infracomunidades de helmintos das duas esp?cies de roedores nos diferentes tamanhos de fragmentos e PARNASO, podendo essas diferen?as estarem relacionadas com as altera??es ambientais causadas pela fragmenta??o que podem produzir tanto efeitos positivos como negativos na transmiss?o dos helmintos parasitos.

Submitted by Sandra Pereira (srpereira@ufrrj.br) on 2018-08-20T14:11:07Z No. of bitstreams: 1 2010 - JOICE DOS SANTOS LIMA BERNARDO.pdf: 3739679 bytes, checksum: 8843284b9fe9600f0ceb730fdda3f94d (MD5) / Made available in DSpace on 2018-08-20T14:11:07Z (GMT). No. of bitstreams: 1 2010 - JOICE DOS SANTOS LIMA BERNARDO.pdf: 3739679 bytes, checksum: 8843284b9fe9600f0ceb730fdda3f94d (MD5) Previous issue date: 2010-04-09 / CAPES / The genus Ctenomys Blainville, 1826 has a systematic and complex although there are morphological and molecular studies on the genus, it is still difficult to identify and classify these individuals who have a high diversity of karyotypes with diploid numbers ranging from 2n = 10 to 2n = 70. However, some authors confirm the usefulness of taxonomic structures such as penis and baculum in the systematic study of rodents. The penis, as well as the baculum (penis bone), has been the subject of several studies because of its great morphological diversity. This work is based on the study of external morphology of the glans penes and baculum of Ctenomys minutus, Ctenomys flamarioni and Ctenomys torquatus to identify possible variations and compare the differences. The study was done from the penis of adult set of the species listed above. Photographs were taken, dissections, drawings with a camera lucida and diaphanization the penis. For analysis of the microstructure of the surface of the glans penes was used the technique of scanning electron microscopy (SEM). The results showed great differences in morphological structure of the baculum of the three species analyzed and showed structures similar to thorns that covered the entire surface of the glans penes with varying distribution over the glans. Our results indicate that the baculum may be an excellent marker complex taxonomic groups such as the Ctenomys. Morphological studies of the surface of the glans penes and baculum combined with genetic studies, molecular and biogeographical can provide important information to better understand the systematic, evolution and taxonomy of the genus. / O g?nero Ctenomys Blainville, 1826 possui uma sistem?tica complexa e apesar de j? existirem estudos moleculares e morfol?gicos sobre o g?nero, ainda ? dif?cil identificar e classificar estes indiv?duos que possuem uma alta diversifica??o cariot?pica com n?meros dipl?ides variando de 2n= 10 a 2n= 70. Por?m, alguns autores confirmam a utilidade taxon?mica de estruturas como p?nis e baculum no estudo da sistem?tica de roedores.O p?nis, assim como o baculum (osso peniano), tem sido alvo de diversos estudos devido sua grande diversidade morfol?gica. O presente trabalho se baseia no estudo da morfologia externa do glans penes e do baculum de Ctenomys minutus, Ctenomys flamarioni e Ctenomys torquatus com o objetivo de identificar poss?veis varia??es e comparar as diferen?as encontradas. O estudo foi feito a partir dos p?nis fixados de indiv?duos adultos das esp?cies listadas anteriormente. Foram feitas fotografias, dissec??es, desenhos com c?mara clara e diafaniza??o dos p?nis. Para an?lises da microestrutura da superf?cie do glans penes foi utilizado a t?cnica de Microscopia Eletr?nica de Varredura (MEV). Os resultados revelaram grandes diferen?as morfol?gicas na estrutura do baculum das tr?s esp?cies analisadas e revelaram estruturas semelhantes a espinhos que recobriam toda a superf?cie do glans penes com distribui??o variando ao longo da glande. Nossos resultados indicam que o baculum pode ser um excelente marcador taxon?mico em grupos complexos como ? o caso dos Ctenomys. Estudos morfol?gicos da superf?cie do glans penes e do baculum aliados a estudos gen?ticos, moleculares e biogeogr?ficos podem fornecer informa??es importantes para melhor entendermos a sistem?tica, evolu??o e taxonomia do g?nero.

There is currently a decline in large, old trees within many ecosystems where they play important ecological and economic roles. One ecosystem suffering from this decline is the bushveld savanna of South Africa. One particularly important species in decline is the ecologically, economically, and culturally significant marula tree (Sclerocarya birrea subsp. caffra). This species’ decline is characterized by a steep drop in recruitment of seedlings into the population. Rodents are known to influence plant communities across many ecosystems through herbivory of adult plants as well as predation of seeds and seedlings. This research provides a record of rodent species present in the bushveld savanna ecosystem of Balule Nature Reserve where a decline in marula recruitment is taking place. Further, it offers a summary of morphological statistics and natural history traits for these rodents. Morphological statistics can be used for species identification. Reproductive status of individuals can inform researchers about reproductive phenology and how it may be affected by environmental conditions. Finally, trapping for this study took place during an historic regional drought. Trapping success was used to develop a limited window into how reduced precipitation affects rodent abundances. Morphological traits measured coincide with those available in the literature. Reproductive activity for males and females of one species, Namaqua rock mouse (Micaelamys namaquensis ), did not overlap completely, with females not being reproductively active while males were. Individuals captured and identified as chestnut climbing mice (Dendromus mystacalis) had statistically significantly longer tails and hind feet, and were heavier than reported in reference works.

Orientador: Bruno Cesar Schimming / Resumo: Os estudos anatômicos aplicados ao conhecimento médico veterinário, como às áreas de cirurgia, anestesia e diagnóstico por imagem são cruciais para uma correta avaliação e abordagem do animal. As capivaras (Hydrochoerus hydrochaeris), os maiores roedores do mundo, são mamíferos encontrados em todo o território brasileiro. Devido a sua ampla distribuição, necessitam-se descrições detalhadas sobre a espécie, dentre elas sobre sua anatomia craniana. Este estudo visou identificar as estruturas cranianas de capivaras em peças anatômicas e imagens radiológicas. Foram utilizados oito crânios e duas cabeças descongeladas para a identificação osteológica, radiográfica e tomográfica das suas estruturas. Foram realizadas medidas craniométricas para as diferentes partes do crânio e calculados os índices cefálicos. O crânio da capivara pode ser dividido em uma parte cranial (neurocrânio) e uma parte facial (viscerocrânio). As capivaras apresentaram um crânio mais robusto e retangular, alongado rostro-caudalmente, adelgaçado na região nasal e levemente convexo na região parietal. O arco zigomático é expandido e largo, a órbita possui um formato circular, o forame infraorbital é bastante desenvolvido, os meatos acústicos externos e as bulas timpânicas são relativamente pequenos e os processos paracondilares são grandes. Estas características anatômicas são compatíveis com o hábito alimentar e com o modo de vida semi-aquático das capivaras, o que pode ser comparado com características report... (Resumo completo, clicar acesso eletrônico abaixo) / Abstract: The anatomical studies applied to veterinary medical knowledge, such as the areas of surgery, anesthesia and diagnostic imaging are pivotal for a correct evaluation and approach of the animal. The capybaras (Hydrochoerus hydrochaeris), the largest rodents of the world, are mammals found throughout the Brazilian territory. Despite their wide distribution, detailed descriptions about the species are needed, among them about their cranial anatomy. This study aimed to identify the cranial structures in skulls and radiological images in the capybaras. Eight skulls and two thawed heads were used for the osteological, radiographic and tomographic identification of their structures. Craniometric measurements were performed for the different parts of the skull and the cephalic indexes were calculated. The skull of the capybara can be divided into a cranial part (neurocranium) and a facial part (viscerocranium). The capybaras had a more robust and rectangular skull, elongated face-caudally, thinned in the nasal region and slightly convex in the parietal region. The zygomatic arch is expanded and wide, the orbit has a circular shape, the infraorbital foramen is well developed, external acoustic meatus and tympanic bullae are relatively small and the paracondillar processes are large. These anatomical characteristics are compatible with the eating habit and semiaquatic life of capybaras, which can be compared with characteristics reported for animals of similar habits. The radiographic i... (Complete abstract click electronic access below) / Mestre

The relationship between alcohol consumption and traumatic brain injury (TBI) often focuses on alcohol consumption increasing the likelihood of incurring a TBI, rather than alcohol use outcomes after TBI. This focus is in part due to the large numbers of TBI patients visiting emergency rooms notable levels of alcohol in their blood. Additionally, increases in alcohol use disorders following TBI can be predicted by previous history of alcohol use. However, studies have also shown patients without a history of an alcohol use disorder can experience increases in problem drinking after single or multiple TBIs. Due to the diffuse impact of alcohol consumption and mild TBI on the brain, it is likely that an interaction exists between TBI outcomes and problematic alcohol use after TBI. To examine the impact of mild repetitive TBI (rmTBI) on voluntary alcohol consumption, male mice were subjected to four mild TBI or sham procedures over a two week period, then offered ethanol (20% v/v) for 2, 4, 6 or 8 weeks using the two-bottle choice, drinking in the dark paradigm. Following the drinking period, mice were sacrificed and brains were extracted to examine expression of the pro-inflammatory cytokine TNF-α, a possible shared mechanism of neuronal damage. An additional cohort of mice was subjected to the same rmTBI and voluntary ethanol paradigm and tested for cognitive and behavioral deficits following the set drinking period. Results indicate there is a temporary decrease in ethanol consumption following rmTBIs compared to Sham mice in this model. Results also suggest an attenuated expression of TNF-α in rmTBI, ethanol drinking groups compared to ethanol exposed mice after the Sham procedure. The outcomes of the cognitive and behavioral tasks suggest that ethanol consumption after rmTBI can cause transient cognitive dysfunction and increased novelty preference.

<p>Landscape effects on species performance currently receives much attention. Habitat loss and fragmentation are considered major threats to species diversity. Deciduous forests in southern Sweden are previous wooded pastures that have become species-rich communities appearing as islands in agricultural landscapes, varying in species composition. <i>Actaea spicata</i> is a long-lived plant occurring in these forests. In 150 populations in a 10-km² area, I studied pre-dispersal seed predation, seed dispersal and pollination. I investigated spatio-temporal dynamics of a tritrophic system including<i> Actaea</i>, a specialist seed predator, <i>Eupithecia immundata</i>, and its parasitoids. In addition, effects of biotic context on rodent fruit dispersal and effects of flowering time and flower number on seed set, seed predation and parasitization were studied. Insect incidences of both trophic levels were related to resource population size and small <i>Eupithecia</i> populations were maintained by the rescue effect. There was a unimodal relationship between seed predation and plant population size. Seed predator populations frequently went extinct in small plant populations, resulting in low average seed predation. Parasitoids were present in large plant populations but did not affect seed predator density. Seed predators aggregated at edges, relaxing seed predation in patch interiors. Flowering phenology was unrelated to seed set and insect incidence. A higher flower number did not influence seed predation but was associated with higher seed set and a tendency for a higher parasitization rate. In the study on fruit dispersal more fruits were removed inside than outside populations. Within plant populations more fruits were removed from large aggregations. Overall, this thesis underlines the importance of plant-animal interactions during different phases of the life cycle. The spatial configuration of host plants plays an important role for the outcome of plant-animal interactions and trophic cascades.</p>

Landscape effects on species performance currently receives much attention. Habitat loss and fragmentation are considered major threats to species diversity. Deciduous forests in southern Sweden are previous wooded pastures that have become species-rich communities appearing as islands in agricultural landscapes, varying in species composition. Actaea spicata is a long-lived plant occurring in these forests. In 150 populations in a 10-km2 area, I studied pre-dispersal seed predation, seed dispersal and pollination. I investigated spatio-temporal dynamics of a tritrophic system including Actaea, a specialist seed predator, Eupithecia immundata, and its parasitoids. In addition, effects of biotic context on rodent fruit dispersal and effects of flowering time and flower number on seed set, seed predation and parasitization were studied. Insect incidences of both trophic levels were related to resource population size and small Eupithecia populations were maintained by the rescue effect. There was a unimodal relationship between seed predation and plant population size. Seed predator populations frequently went extinct in small plant populations, resulting in low average seed predation. Parasitoids were present in large plant populations but did not affect seed predator density. Seed predators aggregated at edges, relaxing seed predation in patch interiors. Flowering phenology was unrelated to seed set and insect incidence. A higher flower number did not influence seed predation but was associated with higher seed set and a tendency for a higher parasitization rate. In the study on fruit dispersal more fruits were removed inside than outside populations. Within plant populations more fruits were removed from large aggregations. Overall, this thesis underlines the importance of plant-animal interactions during different phases of the life cycle. The spatial configuration of host plants plays an important role for the outcome of plant-animal interactions and trophic cascades.

Damage to visual pathways can lead to irreversible blindness. Posterior visual pathways, located within a watershed area, are predisposed to hypoperfusion/reperfusion injury. In a novel rat model of bilateral common carotid artery occlusion (BCCAO), oxidative injury to the superior colliculus (SC), a major visual center within the watershed area was evaluated, in addition to its effects on retinal ganglion cells (RGCs). Nitrotyrosine, a footprint of peroxynitrite-mediated oxidative injury in the SC, and microtubule-associated protein 2, a dendrite marker in the retina, were assessed using immunofluorescence and confocal microscopy. Nitrotyrosine-immunoreactivity in the SC was increased 2 weeks after BCCAO compared to controls. Microtubule-associated protein 2-immunoreactivity in the central inner plexiform layer was reduced 3 weeks after BCCAO compared to controls. Global incomplete cerebral hypoperfusion/reperfusion induced oxidative injury in the SC and retrograde RGC dendritic changes. This suggests that cerebrovascular injury affecting the posterior visual pathways may contribute to vision loss in patients.