O treinamento físico aeróbio crônico regular provoca importantes adaptações autonômicas e hemodinâmicas que influenciam o sistema cardiovascular. O presente trabalho teve como objetivo estudar os efeitos do treinamento físico aeróbio crônico, na citoarquitetura alveolar dos pulmões de ratos Wistar, sob os aspectos morfoquantitativo e funcional, bem como avaliar o efeito do treinamento na modulação da freqüência cardíaca (FC), da pressão arterial (PA), no perfil bioquímico (lactato, hemogasometria, glicemia e perfil lipídico: triglicérides, colesterol total, colesterol fracionado). Desta forma foram utilizados quinze ratos Wistar machos, com idade de 60 dias, que receberam ração própria para roedores e água sem restrição, e foram divididos em três grupos, sendo eles: Controle etário (CE, n=5), saudável sedentário (SS, n=5) e saudável treinado (ST, n=5). Após os registros de todos os parâmetros hemodinâmicos supracitados, os animais foram eutanasiados e fixados por perfusão com solução de formoldeído (4%), sendo o mesmo fixador instilado pela traquéia e perfundido via tronco pulmonar. Métodos estereológicos foram empregados para estimar o número total de aberturas alveolares pulmonares, bem como o volume do pulmão (Princípio de Cavalieri). A pressão arterial sistólica (PAS) foi 140,87 mmHg para o CE, 146,70 mmHg para o SS e 149,53 mmHg para o ST (p = 0,137) A pressão arterial diastólica (PAD) foi 115,83 mmHg para o CE, 107,00 mmHg para o SS e 105,39 mmHg para o ST (p = 0,0004). A pressão arterial média (PAM) foi 128,93 mmHg para o CE, 125,22 mmHg para o SS e 125,34 mmHg para o ST (p = 0,332). A FC foi 388,40 bpm para o CE, 379,83 bmp para o SS e 370,85 bpm para o ST (p = 0,381). O volume do pulmão foi 1,05 cm³ para o CE, 1,24 cm³ para o SS e 0,95 cm³ para o ST (p = 0,484). O número de aberturas alveolares pulmonares foi 21x106 para o CE, 25x106 para o SS e 38x106 para o ST (p = 0,02). Os resultados desse estudo sugerem que o treinamento físico aeróbio crônico em ratos promove um melhor estado hemodinâmico e um aumento no número de aberturas alveolares do pulmão dos animais exercitados. / The cardiovascular system is subject to important autonomic and haemodynamic adaptation by virtue of regular physical training. The aim of this study was to describe the effects of a treadmill physical training on the alveolar cytoarchitecture of the rat\'s lung from a morphoquantitative and functional point of view. Additional aims were to quantify the heart rate (HR), arterial pressure (AP), lactate, arterial blood gases, glucose, triglycerides, total and fractional cholesterol. For this purpose fifteen 60-day-old male Wistar rats were used. Animals were divided into three groups: age control (CE, n=5), sedentary (SS, n=5) and trained rats (ST, n=5). After recording the haemodynamic parameters, the animals were euthanised and perfusion-fixed with a 4 % formaldehyde solution, which was also intratracheally instillated. Design-based stereological methods were applied for estimating the total number of alveolar openings as well as the lung volume using Cavalieri\'s principle. The systolic arterial pressure (SAP) was 140.87 mmHg for the CE, 146.7 mmHg for the SS and 149.53 mmHg for the ST group (p=0.137). The diastolic arterial pressure (DAP) was 115.83 mmHg for the CE, 107 mmHg for the SS and 105.39 mmHg for the ST group (p=0.0004). The mean arterial pressure (MAP) was 128.93 mmHg for the CE, 125.22 mmHg for the SS and 125.34 mmHg for the ST group (p=0.332). HR was 388.4 bpm for the CE, 379.83 bmp for the SS and 370.85 bpm for the ST group (p=0.381). The lung volume was 1.05 cm³ for the CE, 1.24 cm³ for the SS and 0.95 cm³ for the ST group (p=0.484). The total number of alveolar openings alveolar was 21x106 for the CE, 25x106 for the SS and 38x106 for the ST group (p=0.02). The main findings of this research suggest that low-intensity treadmill physical training improves the haemodynamic parameters and increases the total number of alveolar openings of trained rats.

Neste estudo, foram abordados os aspectos quantitativos do gânglio cervical cranial (GCC) esquerdo de preás (Galea spixii spixii) machos, oriundos do criatório da Universidade Federal Rural do Semi-Árido Nordestino de Mossoró-RN. Os seguintes parâmetros foram estimados: volume do GCC, número total e tamanho dos neurônios (uni e binucleados) durante dois períodos do desenvolvimento pós-natal (maturação e envelhecimento). Para tanto, utilizamos quatro grupos etários distintos de animais: neonatos, jovens, adultos e senis. Após a eutanásia dos animais, os GCCs foram fixados (perfusão sistêmica) com solução de formoldeído (4%) em PBS, embebidos em solução de ágar (10%) e seccionados sistemática, uniforme e aleatoriamente, aplicando-se o princípio de amostragem e contagem do fractionator. Os principais resultados desta pesquisa foram: houve diferença significativa entre os grupos etários para os parâmetros: Peso Neonato: 0,00056 ; Jovem: 0,00084; Adulto: 0,00116; Senil: 0,00118. Comprimento Neonato: 1,294 ; Jovem: 2,076; Adulto: 2,304; Senil: 2,082. Largura Neonato: 0,842; Jovem: 1,028; Adulto: 1,062; Senil: 1,174. Volume ganglionar Neonato: 0,34 mm3 ; Jovem: 0,30 mm3; Adulto: 0,39 mm3; Senil: 0,63 mm3 e Volume neuronal médio de neurônios uninucleados Neonato: 2916,90 mm3; Jovem: 3550,00 mm3; Adulto: 7409,00 mm3; Senil: 6701,00 mm3. As principais conclusões deste estudo foram: (i) peso, comprimento e largura do GCC apresentaram aumento significativo de 185%, 60% e 79% respectivamente, durante o período de maturação, enquanto durante o período de envelhecimento, o peso do GCC apresentou queda de 33%, o comprimento um pequeno decréscimo de 10%, entretanto a largura, durante este mesmo período, apresentou um aumento progressivo de mais de 10%, tais alterações relacionadas com a idade refletiram no volume ganglionar, causando hipertrofia; (ii) Essa hipertrofia do GCC de preás é secundária ao desenvolvimento pós-natal (maturação e envelhecimento), aumentando em 85%, acompanhado por um aumento alométrico, da ordem de 6 vezes em relação à massa corporal, aproximadamente 622%; (iii) Há hipertrofia dos neurônios uninucleados que é secundária ao desenvolvimento pós-natal do GCC de preás, aumentando em 154% durante o período de maturação com uma pequena queda de 10% durante o envelhecimento. Finalmente, sugere-se que estudos futuros possam investigar se o tamanho dos territórios de inervação destes gânglios pode desencadear mudanças na micro e macroestrutura dos mesmos, incluindo a possibilidade de ocorrência de neurogênese, que seria estudada por meio de imunomarcadores específicos. / This study aimed at detecting whether post-natal development would exert any effect on the size and number of cranial cervical ganglion (CCG) neurons of male preas (Galea spixii spixii). Twenty left CCGs from twenty preas were harvested from the Animal Facility of the Universidade Federal Rural do Semi-Árido Nordestino, Mossoró- RN and were divided into four different age groups: newborn, young, adult and aged subjects. The following parameters were estimated using Cavalieri\'s principle and optical fractionator and planar rotator , respectively: the volume of CCG, total number and mean volume of uni and binucleate neurons. After euthanasia, the CCGs were perfusion-fixed with a 4% formaldehyde solution in PBS, agar-embedded and, SUR vibrosectioned. The ganglion volume was 0.34 mm3 (newborn), 0.30 mm3 (young), 0.39 mm3 (adult) and 0.63 mm3 (aged animals) (p= 0,012). The mean volume of uninucleate neurons was 2,917 mm3 (newborn), 3,550 mm3 (young), 7,409 mm3 (adult) and 6,701 mm3 (aged animals) (p=0,0001). The main conclusions of this study were: (i) The CCG hypertrophy - 85% - is the result of post-natal development (maturation and ageing), (ii) a 154% increase (hypertrophy) in the volume of uninucleate neurons during the maturation is followed by a 10% atrophy of them during ageing. Future studies may investigate whether the size of the CCG\'s target-organs can affect the structural foundation of CCG and, therefore, add further fuel to the notion that a post-natal neurogenesis may indeed exist in sympathetic ganglia.

A Distrofia Muscular de Duchenne (DMD) é uma doença recessiva ligada ao cromossomo ?X?, causada pela ausência da proteína distrofina que ocorre em vários tecidos, sendo caracterizada por uma severa disfunção da musculatura esquelética ocasionando morte prematura do paciente. Embora controverso, alguns autores reportaram que o GH (hormônio do crescimento) estaria implicado no desenvolvimento da doença e poderia ser utilizado no tratamento da mesma. Desta forma, neste estudo a dosagem sérica de IGF-I (Fator I de crescimento Similar à insulina) um peptídeo GH-dependente que regula as ações do hormônio do crescimento, foi realizada no intuito de verificar se existe correlação ou não entre o desenvolvimento da doença e a concentração sérica de IGF-I. As variações nos níveis deste hormônio foram demonstradas com o decorrer da idade sendo que, nos três primeiros meses todos os animais apresentaram comportamento semelhante com aumento dos níveis de IGF-I, porém no quarto mês os animais Distróficos benignos apresentaram redução média de 34% deste hormônio, enquanto ocorreu aumento de 1% no animal não Distrófico. Ainda, as células granulares adenohipofisárias relacionadas à produção do hormônio de crescimento em cães Golden Retriever com Distrofia Muscular apresentam-se maiores do que nos cães Golden Retriever não distróficos quanto aos seguintes parâmetros (eixo longo, área seccional e volume celular). À microscopia eletrônica de transmissão observamos que as células estudadas apresentam grânulos elétron-densos de parede dupla e distribuídos por todo o citosol. Estes grânulos apresentaram-se maiores nos animais Distróficos quando comparados aos animais não Distróficos. / The Duchenne Muscular Dystrophy (DMD) is a X linked recessive disease, caused by the absence of dystrophin which is found in a variety of tissues and characterized by a severe disfunction of the skeletal musculature that results in a premature death of the patient. Theoretically, the growth hormone (GH) is considered to be associated to the development of Muscular Dystrophy and that could be used in its treatment. Hence in this study, a IGF-I (Insulin like growth factor-I) seric dosage was performed to verify whether or not there might be a between the evolution of the disease and IGF´s seric concentration. IGF-I is a GH-peptide dependent that regulates the GH actions during the growth. Changes in IGF-I levels were recorded during the dog?s post-natal development. On the first trimester, all animals presented similar IGF-I levels, although in the fourth month, a stark 34% decrease was observed in the dystrophic animals whereas a 1% increase was seen in the healthy dog. Furthermore, the GRMD´s granule-containing cells were larger when compared to the healthy animals. The following parameters in this comparison: long axis, cross-sectional area and cell volume. The ultrastructural study showed electron-dense granules composed by a double membrane and homogeneously distributed through the cell. These granules were larger in the dystrophic animals then in healthy dogs.

A seleção de linhagens de frangos de corte com altas taxas de crescimento e aumento de rendimento muscular vem contribuindo para o surgimento de perdas significativas na indústria avícola em decorrência de modificações anatômicas. Dentre estas alterações, destaca-se uma miopatia que se caracteriza pelo aparecimento de graus de estrias esbranquiçadas no músculo Pectoralis major, as quais seguem a direção da fibra muscular, referida como white striping. A etiologia da white striping ainda é desconhecida, mas avaliações histológicas demonstraram que esta alteração é usualmente associada à degeneração muscular e alterações miopáticas. Problemas de qualidade da carne podem ser resultantes de mudanças estruturais, morfométricas e bioquímicas do tecido muscular, com consequente alteração nas fibras musculares. Para tanto este trabalho, conduzido através de dois experimentos, teve como objetivos caracterizar histopatologicamente a miopatia white striping em diferentes graus de severidade (NORM, MOD, SEV) no músculo Pectoralis major de frangos de corte e as características morfométricas das fibras musculares, bem como avaliar a incidência desta condição em frangos de corte de linhagem Cobb abatidos aos 42 dias de idade. No primeiro experimento, a avaliação histopatológica demonstrou uma miopatia degenerativa nos graus de severidade classificados como MOD e SEV, entretanto as amostras de grau MOD apresentaram necrose ou necrose multifocal moderada, enquanto as de grau SEV apresentaram necrose difusa e acentuada, sugerindo avanço do processo degenerativo. Não foi observada proliferação de tecido conjuntivo em nenhum dos graus de white striping analisados, caracterizando este caso como de ocorrência aguda. No segundo experimento, foram verificados menor área de tecido muscular, menor diâmentro e maior densidade de fibras nos músculos Pectoralis major de frangos acometidos com white striping (MOD e SEV) quando comparado aos peitos sem esta condição (NORM). Conforme os resultados obtidos nesta pequisa, pode-se concluir que as fibras com os graus MOD e SEV de white striping demonstraram o início de um processo de regeneração muscular, de característica hiperplásica. / Genetic selection of broiler chickens for growth rate and increase of muscle yield has contributed to the appearance of significant losses in the poultry industry caused by anatomical changes. Among these alterations stands out a myophathy characterized by the occurrence of white striations on Pectoralis major muscle, seen parallel to the direction of muscle fibers, referred as white striping. The etiology of white striping is still unknown but histological changes demonstrated that this alteration is usually associated with muscle degeneration and myopatic changes. Meat quality problems may result from structural, morphological and biochemical changes of muscle tissue with consequent alterations in muscle fibers. To this study, two experiments were conducted, and aimed to histopathologically characterize the myopathy white striping at varying degrees of severity (NORM, MOD, SEV) on broilers Pectoralis major muscle and the morphometric muscle fibers characteristics as well as evaluate the incidence of this condition on Cobb 500 broiler breast fillets slaughtered at 42 days of age. The histopathological evaluation on experiment 1 showed a degenerative myopathy in both MOD and SEV degrees, however the samples classified as MOD showed microscopically necrosis or necrosis multifocal moderate, while the classified as SEV showed necrosis diffuse and strong suggesting an advancement in the degenerative process. The proliferation of connective tissue was not observed for either of the degrees of white striping investigated, characterizing this profile as an acute occurrence. In experiment 2, small area and diameter and higher densities of muscle fiber were observed in broilers Pectoralis major muscle with white striping (MOD and SEV) compared to NORM. According to the results, we can conclude that muscle fibers with white striping showed the beginning of a muscle regeneration process with hyperplasic characteristics.

Machos adultos de P. lineatus tiveram suas gônadas processadas de acordo com as rotinas de microscopia de luz e microscopia eletrônica de transmissão. Para a cultura de células, os testículos foram digeridos enzimaticamente, a suspensão testicular foi separada por gradiente descontínuo com Percoll seguido pelo plaqueamento diferencial por adesão e as células foram cultivadas. Foi realizado o método de enriquecimento das espermatogônias por citometria de fluxo. Os testículos apresentam as regiões anterior, média e posterior com distribuição semelhante dos tipos de células, e o diâmetro nuclear das células germinativas diminui significativamente durante a espermatogênese. As espermatogônias cultivadas por 15 dias com meio para proliferação celular resultaram em grandes aglomerados celulares que foram caracterizados com o anti-Vasa, anti-GFRa1 e anti-OCT4. As culturas que receberam o meio para diferenciação celular mostraram processo de proliferação lento das espermatogônias primárias comparado com a cultura que teve o meio indicado para proliferação celular. / Adult males of P. lineatus had their gonads used according routines of light microscopy and transmission electron microscopy. For cell culture, the testes were enzymatically digested; testicular suspension was separated by discontinuous gradient with Percoll followed by adhesion differential plating and the cells were cultured. The enrichment of the spermatogonia was carried by flow cytometry. The testes present three regions with similar distribution of cell types, and nuclear diameter of germ cells decreases significantly during spermatogenesis. The spermatogonia cultured for 15 days with medium for cell proliferation, resulted in large cell agglomerates which were characterized with the antibodies anti-Vasa, anti-GFRa1 and anti-anti-OCT4. The cultures that receiving medium for cell differentiation showed slow proliferation process of primary spermatogonia compared to cell culture medium suggestive for cell differentiation.

No description available.

Fractures and fracture networks govern the mechanical and fluid flow behavior of rock masses. Tunneling and other rock mechanics applications therefore require the characterization of rock fractures based on geological data. Field investigations produce only a limited amount of data from boreholes, outcrops, cut slopes, and geophysical surveys. In tunneling, the process of excavation creates a priceless opportunity to gather more data during construction. Typically, however, these data are not utilized due to the impedance of sampling and analysis on the flow of construction, and safety concerns with sampling within unlined tunnel sections. However, the use of this additional data would increase the overall safety, quality, and cost savings of tunneling. This study deals with several aspects of the above, with the goal of creating methods and tools to allow engineers and geologists to gather and analysis fracture data in tunnels without interrupting the excavation and without compromising safety. Distribution-independent trace density and mean trace length estimators are developed using principles of stereology. An optimization technique is developed utilizing Differential Evolution to infer fracture size and shape from trace data obtained on two or more nonparallel sampling planes. A method of producing nearly bias free empirical trace length CDF's is also introduced. These new methods and tools were validated using Monte Carlo simulations. A field study was conducted in an existing tunnel allowing the above methods and tools to be further validated and tested. A relational database was developed to aid in storage, retrieval, and analysis of field data. Fracture models were built and updated using fracture data from within the tunnel. Utilization of state of the art imaging techniques allowed for remote sampling and analysis, which were enhanced by the use of 3d visualization techniques. / Ph. D.

The aims of the study were to obtain a method with which it would be possible to produce standardized wounds in the gastric mucosa, and to follow the regeneration of the acid producing parietal cells in those lesions during different experimental conditions. Quantitative methods applied to light and electron microscopy were used. Wounds were cauterized in the corpus mucosa in Sprague-Dawley rats and in addition, pyloroplasty, truncal vagotomy with pyloroplasty or ant- rectoiriy were performed. Other groups of rats with wounds were given long-term treatment with pentagastrin or cimetidine. Stimulation tests were carried out in two groups of wound operated rats. After different periods of time the animals were perfusion fixed and specimens from the wounds and normal mucosa beside the wounds were pre­pared for light and electron microscopy. By means of stereological techniques, different mucosal and cellular structures were then measur­ed. Parietal cells were found in 90 days old wounds. At this stage they were immature with large nuclei and few specialized cell organelles. In spite of this appearance they were able to respond morphologically to stimulation and to secrete acid. With further healing the morphology of the parietal cells became normal, but their volume fraction in the mucosa remained subnormal. The fraction of mucosa occupied by epithel­ial cells also stayed lower than normal. Pyloroplasty resulted in decreased cell and nuclear size of both normal and regenerating parietal cells. In the latter, there was also a de­crease in the mitochondrial volume density. If a truncal vagotomy was added to the pyloroplasty these changes disappeared and, in addition, an increase in parietal cell volume density was noticed in the normal mucosa. Antrectorny produced smaller parietal cells, and their maturation was delayed. Furthermore, mucosal thickness decreased. If pentagastrin was given to rats with wounds an increase in the number of parietal cells was noted, but maturation and morphology remained unaffected. Cimetidine treatment did not affect the parietal cell volume density in wounds or normal mucosa. However, a large increase in the secretory surface density was noticed when the effect of the last dose had ceas­ed. / <p>S. 1-45: sammanfattning, s. 47-121 utgörs av 5 uppsatser</p> / digitalisering@umu

L’exposition du fœtus à l’éthanol est reconnue comme étant la principale cause de maladies évitables lors du développement. Une forte exposition à l’alcool durant la gestation peut occasionner des dysmorphies cranio-faciales et des retards mentaux, ainsi que des troubles d’apprentissages et du comportement. Le développement du système visuel est également perturbé chez une grande majorité d’enfants qui ont été exposés à l’alcool. Lorsque les doses prises sont élevées, le système visuel peut présenter une panoplie de symptômes comme une augmentation de la tortuosité des vaisseaux rétiniens, de la myopie, de l’hypermétropie, du strabisme et une hypoplasie du nerf optique. Cependant, très peu d’études se sont penchées sur les effets de plus faibles doses sur le développement du système visuel du primate. Le singe est un excellent modèle pour étudier le système visuel car il possède plusieurs similitudes avec l’humain tant au niveau développemental qu’au niveau structurel. De plus, le singe utilisé, le Chlrocebus aethiops sabeus, possède l’avantage que des individus de cette espèce ont une consommation naturelle et volontaire à l’alcool. Une étude (Clarren et al., 1990) a suggéré qu’une faible exposition à l’alcool du fœtus du primate non humain occasionnait une diminution du nombre de cellules ganglionnaires de la rétine (CGRs). Étant donné que le corps genouillé latéral dorsal (CGLd) reçoit la plupart de ses intrants de la rétine, il est raisonnable d’assumer que les couches rétino-récipientes du CGLd devraient être aussi affectées. Nous avons alors émis l’hypothèse que le CGLd devrait également subir une diminution du nombre de neurones. Pour la première fois, nous avons utilisé une méthode stéréologique pour quantifier le nombre de cellules dans les couches parvo- (P) et magnocellulaires (M) du CGLd. Contrairement à notre hypothèse de départ, nous n’avons pas observé de diminution dans le nombre global de neurones dans le CGLd des animaux exposés à l’alcool par rapport à des sujets contrôles, ni une diminution de son volume. Nous avons toutefois observé une diminution de la taille du corps cellulaire seulement dans la population M du CGLd. Ces résultats suggèrent que le système visuel est affecté par une faible exposition à l’alcool durant son développement qui devrait se traduire sur le comportement par des déficits dans les fonctions de la voie M. / An alcohol exposition during the gestation is recognized as the leading causes of preventable developmental disorders. A heavy exposure to ethanol can lead to a litany of symptoms ranging from cranio-facial dysmorphology and mental retardation to developmental learning and behavioural disorders. The visual system development is also the target for a high percentage of children exposed prenatally to alcohol. When the concentrations are relatively high, the visual system can show signs of increased tortuosity of the retina vessels, myopia, hyperopia, strabismus and hypoplasia of the optic nerve. Few studies have examined the effects of a moderate exposure to alcohol on the development of the primate visual system. The monkey is an excellent model to study the visual system because this model owns a lot of similarities with human at the developmental and structural levels. Moreover, the monkey used for this study, Chlrocebus sabeus, naturally and voluntarily consumes alcohol. One study (Clarren et al., 1990) suggested that a low dose exposure of alcohol induce a diminution of neurons in the ganglion retinal cell population. But, no information is given about the numbers and the cell population of neurons. We hypothesized that the lateral geniculate nucleus (LGN) should show a decrease in the number of neurons after moderate prenatal exposure to alcohol. The LGN is a key region because it is situated between the region that receives visual information, the retina, and the first visual region that analyzes the visual information, the primary visual cortex (V1). We utilized a stereological method to study the effects of moderate doses of alcohol on the neuronal population of the LGN in the non-human primate. In opposition to our hypothesis, we have not observed any diminution in the number of neurons or the volume of the LGN. However, we have found a diminution in the neuronal volume of only the magnocellular (M) region. These results suggest that the visual system is affected by a low dose of alcohol that should translate into deficits in the functions of the M pathway.

Univerzita Karlova v Praze Přírodovědecká fakulta Studijní program: Vývojová biologie (P1520) Studijní obor: Vývojová biologie (1501V000) Oleksandr Chernyavskiy Zdokonalené metody pro snímání obrazových dat a analýzu tkání a buněk pomocí konfokální a multifotonové mikroskopie Improved Methods of Image Acquisition and Analysis of Tissues and Cells by Confocal and Multi-Photon Microscopy Abstrakt disertační práce Školitel: RNDr. Lucie Kubínová CSc Praha, 2015 Abstract The aim of this study was to develop methods and approaches for image acquisition with subsequent image analysis of data, obtained by confocal and two- photon excitation microscopy as well as their combination, enabling new possibilities of visualization and assessment of information on biological tissues and cell structures in 3D and their measurement. We focused on methods that exploited advantages of confocal and multi-photon excitation microscopy. Our further aim was to demonstrate the applicability of non-invasive approach for in vivo applications, usefulness and the relevance of these methods in several special biological applications with emphasis on improved image acquisition, analysis and evaluation of real biological specimens. The present work was not oriented on just one specific biological problem, but rather to methodological...