Cicatrização da menbrana timpânica na timpanocentese com laser de argônio comparado à técnica com microlanceta : estudo experimental em ratos

Introdução: Otite média secretora (OMS) e otite média aguda recorrente (OMAR) podem necessitar tratamento cirúrgico para adequada ventilação da orelha média. A abertura clássica do tímpano (timpanocentese) requer incisão por microlanceta sob controle de microscópio cirúrgico e mantém-se patente por alguns dias. Estudos recentes sugerem que a timpanocentese feita por diferentes lasers pode permanecer permeável por maior tempo, o que possibilitaria a normalização da otite média. Objetivo: Comparar a técnica de timpanocentese incisional com microlanceta à timpanocentese feita com laser de argônio. Material e métodos: Estudo experimental com 34 ratos linhagem Wistar, albinos, machos adultos pesando cerca de 300g, anestesiados com cetamina 27 mg/kg e xilazina 2,7 mg/kg, e submetidos à timpanocentese incisional com microlanceta no ouvido direito (ML-OD), e à timpanocentese mediada por laser de argônio no ouvido esquerdo (LA-OE). As timpanocentes foram avaliadas periodicamente até a cicatrização. Resultados: Não houve diferença significativa no tempo de cicatrização das timpanocenteses feitas com laser de argônio ou microlanceta. Todas timpanocenteses cicatrizaram dentro de 10 dias. Conclusão: A timpanocentese com laser de argônio apresentou patência e cicatrização semelhantes à técnica clássica com microlanceta realizada em ratos Wistar sem enfermidades de orelha média.

O estudo histológico do osso temporal do ovino : uma contribuição para a caracterização da ovelha como modelo animal para tratamento e investigação experimental em otologia

Resumo não disponível

Avaliação histomorfométrica do efeito do ultrasom pulsado nas falhas ósseas provocadas em fêmures de rato: estudo experimental

SILVA, J. S. P. Avaliação histomorfométrica do efeito do ultrasom pulsado nas falhas ósseas provocadas em fêmures de rato: estudo experimental . 2000. 85 f. Dissertação (Mestrado) – Faculdade de Medicina, Universidade de São Paulo. São Paulo, 2000.

Análise do efeito de polimorfismos não-sinônimos em genes de reparo de DNA da via BER na resposta inflamatória da meningite

In vitro and in animal models, APE1, OGG1, and PARP-1 have been proposed as being involved with inflammatory response. In this work, we have investigated if the SNPs APE1 Asn148Glu, OGG1 Ser326Cys, and PARP-1 Val762Ala are associated to meningitis and also developed a system to enable the functional analysis of polymorphic proteins. Patients with bacterial meningitis (BM), aseptic meningitis (AM) and controls (non-infected) genotypes were investigated by PIRA-PCR or PCR-RFLP. DNA damages were detected in genomic DNA by Fpg treatment. IgG and IgA were measured from plasma and the cytokines and chemokines were measured from cerebrospinal fluid samples using Bio-Plex assays. The levels of NF-κB and c-Jun were measured in CSF by dot blot assays. A significant (P<0.05) increase in the frequency of APE1 148Glu allele in BM and AM patients was observed. A significant increase in the genotypes Asn/Asn in control group and Asn/Glu in BM group was also found. For the SNP OGG1 Ser326Cys, the genotype Cys/Cys was more frequent (P<0.05) in BM group. The frequency of PARP-1 Val/Val genotype was higher in control group (P<0.05). The occurrence of combined SNPs increased significantly in BM patients, indicating that these SNPs may be associated to the disease. Increasing in sensitive sites to Fpg was observed in carriers of APE1 148Glu allele or OGG1 326Cys allele, suggesting that SNPs affect DNA repair activity. Alterations in IgG production were observed in the presence of SNPs APE1Asn148Glu, OGG1Ser326Cys or PARP-1Val762Ala. Reductions in the levels ofIL-6, IL-1Ra, MCP-1/CCL2and IL-8/CXCL8 were observed in the presence of APE1148Glu allele in BM patients, however no differences were observed in the levels of NF-κB and c-Jun considering genotypes and analyzed groups. Using APE1 as model, a system to enable the analysis of cellular effects and functional characterization of polymorphic proteins was developed using strategies of cloning APE1 cDNA in pIRES2-EGFP vector, cellular transfection of the construction obtained, siRNA for endogenous APE1 and cellular cultures genotyping. In conclusion, we obtained evidences of an effect of SNPs in DNA repair genes on the regulation of immune response. This is a pioneering work in the field that shows association of BER variant enzymes with an infectious disease in human patients, suggesting that the SNPs analyzed may affect immune response and damage by oxidative stress level during brain infection. Considering these data, new approaches of functional characterization must be developed to better analysis and interactions of polymorphic proteins in response to this context

Comparação entre critérios de seleção de precocidade sexual e a associação destes com características de crescimento em bovinos Nelore

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Dois critérios de seleção na pré-desmama em bovinos da raça Gir. 1. Estimativas de parâmetros genéticos

Dados de 16.592 animais da raça Gir, provenientes do controle de desenvolvimento ponderal da Associação Brasileira dos Criadores de Zebu, nascidos no período de 1978 a 1994, criados em diversas condições de ambiente no Brasil, foram usados para estimar parâmetros genéticos e ambientais das características ganho médio diário do nascimento à desmama (GMD) e dias para atingir 160 kg do nascimento à desmama (D160). As estimativas dos componentes de variância e herdabilidade foram obtidas pelo método da máxima verossimilhança restrita por modelos animais uni-característica. Foram utilizados dois modelos o modelo 1, com grupo de contemporâneos (GC) como efeito fixo e com efeitos genético aditivo de animal e materno, efeito de ambiente permanente materno e erro como aleatórios; e modelo 2, com os mesmos efeitos do modelo 1, mas com covariância genética entre os efeitos direto e materno igual a zero (sigmaam=0). As estimativas de herdabilidade, utilizando-se o Modelo 1, foram iguais a 0,11; 0,04; 0,11 e 0,07 para GMD (efeito direto), GMD (efeito materno), D160 (efeito direto) e D160 (efeito materno), respectivamente. Para o Modelo 2, as estimativas de herdabilidade foram: 0,12; 0,05; 0,10 e 0,05 para GMD (efeito direto), GMD (efeito materno), D160 (efeito direto) e D160 (efeito materno). A diferença entre os modelos pela não inclusão da estrutura de covariância entre os efeitos direto e materno (sigmaam=0), neste trabalho, não foi significativa. A contribuição da variância residual (como porcentagem da variação fenotípica total) foi de 76% para GMD e de 80% para D160, para os dois modelos, indicando a necessidade da melhoria do ambiente de criação, assim como a padronização e formação de lotes de manejo, o que reflete na possibilidade de melhor definição de grupo de contemporâneos.

Resposta à Seleção para Características de Desempenho em um Rebanho de Seleção de Suínos

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

Estimativas de parâmetros genéticos para os escores visuais e suas associações com peso corporal em bovinos de corte

Foram avaliados 27.523 e 21.746 registros das características conformação, precocidade e musculatura à desmana e ao sobreano, respectivamente, para estimar os componentes de covariância entre estas características e entre estas e os pesos corporais medidos nas mesmas idades. Para as análises dos dados, foram empregados modelos animais com efeitos genéticos direto e materno e efeito de ambiente permanente materno. Máxima verossimilhança restrita foi empregada para estimar os parâmetros genéticos. As estimativas de herdabilidade dos escores à desmama foram 0,13; 0,25 e 0,23 para conformação, precocidade e musculatura, respectivamente. As estimativas de herdabilidade dos escores visuais avaliados ao sobreano foram de maiores magnitudes (0,24; 0,32 e 0,27 para conformação, precocidade e musculatura, respectivamente). As estimativas das correlações genéticas entre escores medidos às mesmas idades, considerando desmana e sobreano, foram 0,67 e 0,75 entre conformação e precocidade; 0,61 e 0,71 entre conformação e musculatura; 0,95 e 0,95 entre precocidade e musculatura. As correlações genéticas estimadas entre o peso corporal à desmama e conformação, precocidade e musculatura, respectivamente, foram 0,97; 0,67 e 0, 62. As estimativas entre conformação, precocidade, musculatura ao sobreano e o peso corporal foram 0,83; 0,59 e 0,58, respectivamente. Os resultados indicam que os escores visuais podem ser utilizados como critérios de seleção. Aumento nos pesos corporais deve ser esperado como resposta correlacionada à seleção para essas características.

Interferindo com oscilações de alta frequência no hipocampo epiléptico: consequências para as crises espontâneas

Conselho Nacional de Desenvolvimento Científico e Tecnológico

Caracterização comportamental e distribuição de neurônios inibitórios em um modelo animal de autismo induzido por ácido valpróico

Autism comprises a heterogeneous group of neurodevelopmental disorders that affects the brain maturation and produces sensorial, motor, language and social interaction deficits in early childhood. Several studies have shown a major involvement of genetic factors leading to a predisposition to autism, which are possibly affected by environmental modulators during embryonic and post-natal life. Recent studies in animal models indicate that alterations in epigenetic control during development can generate neuronal maturation disturbances and produce a hyper-excitable circuit, resulting in typical symptoms of autism. In the animal model of autism induced by valproic acid (VPA) during rat pregnancy, behavioral, electrophysiological and cellular alterations have been reported which can also be observed in patients with autism. However, only a few studies have correlated behavioral alterations with the supposed neuronal hyper-excitability in this model. The aim of this project was to generate an animal model of autism by pre-natal exposure to VPA and evaluate the early post-natal development and pre-puberal (PND30) behavior in the offspring. Furthermore, we quantified the parvalbumin-positive neuronal distribution in the medial prefrontal cortex and Purkinje cells in the cerebellum of VPA animals. Our results show that VPA treatment induced developmental alterations, which were observed in behavioral changes as compared to vehicle-treated controls. VPA animals showed clear behavioral abnormalities such as hyperlocomotion, prolonged stereotipies and reduced social interaction with an unfamiliar mate. Cellular quantification revealed a decrease in the number of parvalbumin-positive interneurons in the anterior cingulate cortex and in the prelimbic cortex of the mPFC, suggesting an excitatory/inhibitory unbalance in this animal model of autism. Moreover, we also observed that the neuronal reduction occurred mainly in the cortical layers II/III and V/VI. We did not detect any change in the density of Purkinje neurons in the Crus I region of the cerebellar cortex. Together, our results strengthens the face validity of the VPA model in rats and shed light on specific changes in the inhibitory circuitry of the prefrontal cortex in this autism model. Further studies should address the challenges to clarify particular electrophysiological correlates of the cellular alterations in order to better understand the behavioral dysfunctions

Influência de fatores individuais e sociais sobre as respostas endócrina e comportamental de callithrix jacchus a desafios ambientais físicos e sociais

Regarding the growing number of human beings with physical and mental pathologies associated to different stressor agents, attempts are being made to validate animal models with a close phylogenetic resemblance to man, to study stress response. Callithrix jacchus has been widely used in biomedical research, including on stress, but there is scarce information in the literature about how individual and social factors modulate stressor response in this species. This study uses 4 approaches to investigate the response of male and female adult C. jacchus, under situations of stress, and in the first we show evidence of the importance of this animal as an experimental model in research involving the hypothalamus-pituitary-adrenal axis. And we investigate if sex and baseline cortisol levels modulate the behavioral and hormonal response to separation. In two additional approaches investigate if type of social support (co-specific parent or non-parent) and social rank interfere in behavioral and hormonal when the animal are exposure to a new environment, paired with a co-specific (F2), exposure of the animal to a new environment, isolated (F3) or during reunion (F4). Finally, we also investigated the androgen levels in the males, with a focus on the challenge hypothesis, referring to environmental responsiveness and male-male exposure to relatives and non-relatives of C. jacchus. It was observed that: (1) the baseline cortisol of the animal is predictive of cortisol reactivity at separation; (2) males and females do not show dimorphism in the response of cortisol to stressors, although the females have higher baseline levels of this hormone and exhibit higher frequencies of anxiety-related behaviors; (3) only social support provided by relatives proved to be effective in buffering the cortisol response. In behavioral terms this response was dimorphic, showing that only the male dyads displayed an attenuated response to stress; (4) the males showed differences in cortisol levels as a function of social rank and study phases, whereas in the females no such alterations were observed. The males with indefinite dominance hierarchy (IDH) had reduced cortisol in F2 and F4, while the IDH females showed an increase in F3 and F4; (5) the males of relative and non-relative dyads did not exhibit variations in androgen levels as a function of a new environment. These results, taken together, (a) corroborate the use of C. jacchus as a good animal model for stress-related studies, given that they exhibit similar behavioral and physiological alterations to those of human beings in response to stressor agents; (b) point to the importance of considering individual and social modulating factors during experiments with stressors; (c) provide more reliable comparison parameters in studies where these primates are used as animal models, and (d) show that androgens vary as a function of genetic proximity (relative or non-relative) when the animals are faced with physical and social environmental challenges, thus providing important information for studying the challenge hypothesis in this species

Diferenças funcionais das regiões hipocampais na formação da memória do tipo o quê", "quando" e "onde" em ratos

Episodic memory refers to the recollection of what, where and when a specific event occurred. Hippocampus is a key structure in this type of memory. Computational models suggest that the dentate gyrus (DG) and the CA3 hippocampal subregions are involved in pattern separation and the rapid acquisition of episodic memories, while CA1 is involved in memory consolidation. However there are few studies with animal models that access simultaneously the aspects ‗what-where-when . Recently, an object recognition episodic-like memory task in rodents was proposed. This task consists of two sample trials and a test phase. In sample trial one, the rat is exposed to four copies of an object. In sample trial two, one hour later, the rat is exposed to four copies of a different object. In the test phase, 1 h later, two copies of each of the objects previously used are presented. One copy of the object used in sample trial one is located in a different place, and therefore it is expected to be the most explored object.However, the short retention delay of the task narrows its applications. This study verifies if this task can be evoked after 24h and whether the pharmacological inactivation of the DG/CA3 and CA1 subregions could differentially impair the acquisition of the task described. Validation of the task with a longer interval (24h) was accomplished (animals showed spatiotemporal object discrimination and scopolamine (1 mg/kg, ip) injected pos-training impaired performance). Afterwards, the GABA agonist muscimol, (0,250 μg/μl; volume = 0,5 μl) or saline were injected in the hippocampal subregions fifteen minutes before training. Pre-training inactivation of the DG/CA3 subregions impaired the spatial discrimination of the objects (‗where ), while the temporal discrimination (‗when ) was preserved. Rats treated with muscimol in the CA1 subregion explored all the objects equally well, irrespective of place or presentation time. Our results corroborate the computational models that postulate a role for DG/CA3 in spatial pattern separation, and a role for CA1 in the consolidation process of different mnemonic episodes

Administração repetida de baixas doses de reserpina: um possível modelo para o estudo de déficits cognitivos e motores associados à Doença de Parkinson

Parkinson's disease (PD) is one of the most common neurodegenerative brain disorders and is characterized primarily by a progressive degeneration of dopaminergic neurons nigroestriatais. The main symptoms of this disease are motor alterations (bradykinesia, rigidity, tremor at rest), which can be highly disabling in advanced stages of the condition. However, there are symptomatic manifestations other than motor impairment, such as changes in cognition, mood and sensory systems. Animal models that attempt to mimic clinical features of PD have been used to understand the behavioral and neural mechanisms underlying neurophysiological disturbance of this disease. However, most models promote an intense and immediate motor impairment, consistent with advanced stages of the disease, invalidating these studies for the evaluation of its progressive nature. The administration of reserpine (a monoamine depletor) in rodents has been considered an animal model for studying PD. Recently we found that reserpine (in doses lower than those usually employed to produce the motor symptoms) promotes a memory deficit in an aversive discrimination task, without changing the motor activity. It was suggested that the administration of this drug in low doses can be useful for the study of memory deficits found in PD. Corroborating this data, in another study, acute subcutaneous administration of reserpine, while preserving motor function, led to changes in emotional context-related (but not neutral) memory tasks. The goal of this research was to study the cognitive and motor deficits in rats repeatedly treated with low doses of reserpine, as a possible model that simulates the progressive nature of the PD. For this purpose, 5-month-old male Wistar rats were submitted to a repeated treatment with vehicle or different doses of reserpine on alternate days. Cognitive and motor parameters and possible changes in neuronal function were evaluated during treatment. The main findings were: repeated administration of 0.1 mg / kg of reserpine in rats is able to induce the gradual appearance of motor signs compatible with progressive features found in patients with PD; an increase in striatal levels of oxidative stress and changes in the concentrations of glutamate in the striatum were observed five days after the end of treatment; in animals repeatedly-treated with 0. 1 mg/kg, cognitive deficits were observed only after the onset of motor symptoms, but not prior to the onset of these symptoms; 0.2 mg / kg reserpine repeated treatment has jeopardized the cognitive assessment due to the presence of severe motor deficits. Thus, we suggest that the protocol of treatment with reserpine used in this work is a viable alternative for studies of the progressive appearance of parkinsonian signs in rats, especially concerning motor symptoms. As for the cognitive symptoms, we suggest that more studies are needed, possibly using other behavioral models, and / or changing the treatment regimen

Caracterização comportamental endócrina das fases ontogenéticas de sagüi comum (Callithrix jacchus)

The use of animal models in biomedical research is ever increasing. Models that use primates might also have advantages in terms of low maintenance costs and availability of biological knowledge, thereby favoring their use in different experimental protocols. Many current stress studies use animal models at different developmental stages since biological response differs during ontogeny. The aims of this study were to perform a detailed characterization of the developmental stages of common marmosets (Callithrix jacchus), a very important animal model used in biomedical research. Ten subjects, 6 females and 4 males, were followed from birth to initial adult age (16 months). Behavioral and fecal collection for measurement of adrenal (cortisol) and sex (progesterone, estradiol and androgens) hormones took place twice a week during the first month of life and once a week for the remainder of the study. Behavior was observed for 30 minutes in the morning (0700-09:00h) and afternoon (12:00-14:00h). Behavioral profile showed changes during ontogeny, characterizing the 4 developmental stages and the respective phases proposed by Leão et al (2009).. Differentiation of developmental stages was considered using the onset, end, change and stabilization of the behavioral profile parental care (weaning and carrying), ingestion (solid food), affiliation (social grooming) and autogrooming, agonism (scent marking and piloerection) and play behavior and endocrine profile. Infant weaning and carrying terminated within the infantile stage and the peak of solid food ingestion was recorded in the infantile III phase. Receiving grooming was recorded earlier than grooming performed by the infant and autogrooming. The first episode of scent marking was recorded in the 4th week and it was the least variable behavior, in terms of its onset, which, in almost all animals, was between the 5th and 7th week of life. Solitary play and play with the twin started around the 7th week and play with other members of the group started 8 weeks later. Sex hormone secretion started to differ from basal levels between the 21st and 23rd week of life, in males and females, suggesting that puberty occurs simultaneously in both sexes. Basal cortisol, even at an early age, was higher in females than in males. However, cortisol was not correlated with the juvenile stage, as expected, since this stage corresponds to the transition between infancy and adult age and most behaviors are intensified by this time. The behavioral and endocrine profile of subadult animals did not differ from that of the adults. These results provide more detailed parameters for the developmental process of C. jacchus and open new perspectives for the use of experimental approaches focused on the intermediate ontogenetic phases of this species

Estudo da organização funcional do sistema circadiano por meio de ferramentas computacionais e matemáticas

Circadian rhythms are variations in physiological processes that help living beings to adapt to environmental cycles. These rhythms are generated and are synchronized to the dark light cycle through the suprachiasmatic nucleus. The integrity of circadian rhythmicity has great implication on human health. Currently it is known that disturbances in circadian rhythms are related to some problems of today such as obesity, propensity for certain types of cancer and mental disorders for example. The circadian rhythmicity can be studied through experiments with animal models and in humans directly. In this work we use computational models to gather experimental results from the literature and explain the results of our laboratory. Another focus of this study was to analyze data rhythms of activity and rest obtained experimentally. Here we made a review on the use of variables used to analyze these data and finally propose an update on how to calculate these variables. Our models were able to reproduce the main experimental results in the literature and provided explanations for the results of experiments performed in our laboratory. The new variables used to analyze the rhythm of activity and rest in humans were more efficient to describe the fragmentation and synchronization of this rhythm. Therefore, the work contributed improving existing tools for the study of circadian rhythms in mammals