Lesões na inervação do trato urinário inferior ocasionado por traumatismo raquimedular afetam geralmente o músculo detrusor e o esfíncteres uretrais. Estas alterações acarretam problemas basicamente de incontinência urinária e aumento da pressão intravesical, decorrente deste traumatismo, trazendo consequências para o funcionamento do sistema urinário superior. Quantificar os elementos fibrosos da matriz extracelular e fibras musculares das bexigas neurogênicas hiper-reflexas comparando-as com bexigas normais. Foram utilizadas 6 amostras de bexigas neurogênicas de indivíduos que foram submetidos a cirurgia de reparação por cistoenteroplastia realizados pelo serviço de urologia do Hospital Municipal Souza Aguiar, estas amostras foram fixadas imediatamente em solução tamponada de formalina a 10%. O controle com amostras iguais as do estudo extraída de cadáveres cuja causa morte não relacionava-se ao sistema urogenital macroscópicamente. O material foi submetido as seguintes técnicas histoquímicas: H&E, van Gieson e Resorcina Fucsina resorcina de Weigert com prévia oxidação pela oxona. Imunohistoquímica: anti-elastina. A observação dos cortes corados pelo van Gieson demonstrou uma diminuição significativa do músculo liso de 13% e aumento do colágeno em 72% e as fibras do sistema elástico um aumento de 101%. Conclusão. Nas bexigas neurogênicas hiper-reflexas o músculo detrusor e os elementos fibrosos da matriz foram profundamente modificados. As fibras do sistema elástico foram as mais afetadas. / Lesions on lower urinary tract innervations caused by spinal cord injuries usually affect the detrusor muscle and urethral sphincter. Beside the smooth muscle fibers, the collagen fibers and elastic system fibers, fibrous components of the extracellular matrix of the bladder wall, are strongly related to vesicle bladder compliance. For this reason the aim of this work is to quantify the fibrous elements of the extracellular matrix and muscle fibers of the neurogenic bladder hyperreflexia. Samples of neurogenic bladder were obtained from six men who had previously undergone surgical repair. The control group samples (n=6) were similarly obtained from patients whose deaths were not related to the urogenital system. The samples were stained using the following histochemical techniques: H&E, Van Gieson, Weigert and Sirius Red. Sections stained with Sirius Red were observed under polarization light microscopy to characterize possible different kinds of collagen. Immunohistochemical technique was used to characterize and quantify the elastic system fibers. Quantification analysis was performed by stereological methods. An increase of 72% of the collagen was observed. Nevertheless, the most significant difference observed was the raising of 101% of the elastic system fibers. Contrary the smooth muscle fibers showed a decrease of 13%. In the neurogenic bladder with detrusor hyperreflexia the fibrous elements of the extracellular matrix and smooth muscle fibers were greatly modified. The elastic system fibers seem to be the most affected in this disease.

The heat affected zone (HAZ) of friction stir welded Al 7075 T7 shows diminished corrosion resistance properties when compared with the parent material. Corrosion attack in this region of the weld is primarily intergranular and is associated with the presence of precipitate free zones. Current TEM research conducted at Brigham Young University by Dr. Bin Cai finds a correlation between precipitate free zone (PFZ) width and grain boundary geometry. As both grain boundary geometry and the PFZ are associated with modes of failure in 7XXX aluminum, this paper makes a comparison of grain boundary character distributions (GBCD) in the HAZ and the parent metal via multi-section plane five-parameter stereology. The stereology is conducted in a convenient macroscopic coordinate frame, associated with the HAZ. This is the first investigation to determine the GBCD in the HAZ of friction-stirred weld material and requires multiple section plane sampling. It is discovered that aluminum here exhibits the property of non-sidedness, a long assumed but unproven characteristic. Further comparisons between the two microstructures are conducted relative to (2-dimensional) grain boundary network connectivity, recovered from EBSD data in each section plane. It is shown that the relative fraction of grain boundaries of misorientation character associated with smaller PFZ size is larger in the HAZ as compared to the parent material. A commensurate decrease in the connectivity (radius of gyration) of grain boundaries of character conducive to larger PFZ size is also found in the HAZ, relative to the parent material. Distribution of inclinations changes as a function of grain boundary geometry. Surface area per unit volume of CSL and low angle random (LAR) misorientations increases in the HAZ, while high angle random (HAR) boundaries decrease. In the case of LAR and some CSL boundaries, a reorientation occurs in which macroscopic normals of these interfaces rotate. It is anticipated that these significant changes in the GBCD within the HAZ could be important in terms of understanding the post-weld mechanical and physical properties in friction-stirred materials.

L'utilisation récente des nanoparticules (NPs) comme vecteurs pour l'imagerie et l'adressage d'agents thérapeutiques en nano-médecine nécessite la compréhension de leurs mécanismes d'internalisation et de transport au niveau des barrières biologiques. Dans ce contexte, l'objectif de cette étude est de caractériser l'interaction et la transcytose de NPs de silice fluorescentes en fonction de leur taille (15, 50 et 100 nm) dans un modèle in-vitro de barrière endothéliale pulmonaire humaine. L'internalisation et le transport trans-endothélial des NPs a été analysé quantitativement à l'échelle nanométrique par microscopie électronique à transmission (MET) combinée à de la stéréologie. Un transport trans-endothélial a été observé pour toutes les tailles de NPs. Néanmoins, l'analyse de la distribution intracellulaire révèle une tendance à l'accumulation dans les voies de dégradation cellulaires pour les NPs de 50 et 100 nm. Cette accumulation est moindre pour les NPs de 15 nm. L'internalisation des NPs a également été analysée par cytométrie en flux et MET en présence de différents inhibiteurs de l'endocytose dans le but d'identifier leurs voies d'internalisation. En fonction de la taille des NPs, les mécanismes d'endocytose varient, suggérant une dépendance du transport trans-cellulaire à certains mécanismes d'endocytose. L'internalisation des NPs de 15 nm par la voie d'endocytose cavéole dépendante pourrait ainsi expliquer l'efficacité de leur transport du côté basal. Les méthodologies développées pour l'étude du transport trans-cellulaire des NPs de silice peuvent être appliquées à l'étude de NPs synthétiques plus complexes ou de NPs biologiques, telles que les lipoprotéines de basse-densité, et ce dans un contexte pathologique. / The recent use of nanoparticles (NPs) as carriers for imaging and delivery of therapeutics agents in nanomedecine involves understanding their endocytosis and transcytosis mechanisms at biological barriers. In this context, the aim of this study was to characterize the interaction and transcytosis of fluorescent silica NPs in function of their size (15, 50, and 100 nm) in an in-vitro model of human pulmonary endothelial barrier. NPs internalization and trans-endothelial transport has been quantitatively analyzed at nanometer resolution using transmission electron microscopy (TEM) combined with stereology. Trans-endothelial transport has been observed for each size of NPs. However cellular distribution analysis shows an accumulation in the cellular degradation pathways for 50 nm and 100 nm NPs. Whereas 15 nm NPs are less accumulated. NPs uptake was also analyzed by flow cytometry and TEM in the presence of different inhibitors to decipher NPs internalization pathways. Depending on NPs size, the involved endocytosis pathways were different, suggesting a dependency of trans-cellular transport toward endocytic mechanisms. The specific internalization of 15 nm NPs by the caveolin dependant pathway could explain the efficacy of their release at the basal side. Techniques developed for the study of the trans-cellular transport of silica NPs can also be applied to more complex synthetic NPs or biological NPs, such as low-density lipoproteins, in a pathological context.

<p>Dopamine regulates movement, cognition and the rewarding effects of addictive drugs. Sex differences mediated by gonadal hormones affect each of these processes. An extensive literature suggests that estrogen augments dopaminergic function. Our laboratory found that female rats exhibit increased locomotor stimulation in response to cocaine and greater cocaine-induced dopamine overflow compared to males, sex differences that emerge in early adulthood. Currently, the underlying mechanisms for these differences are poorly understood. I hypothesized that female rats would have more dopamine neurons in midbrain regions and that ovarian hormones would exert trophic effects on dopamine neurons. Immunohistochemical and stereological techniques were used to quantitate the number of cells in the SNpc and VTA of male and female rats and mice to assess: (1) if sex differences in dopamine neuron number exist and when they emerge, (2) how gonadal hormones influence dopaminergic cell number and dopamine-mediated behaviors (3) the role of specific hormone receptors in the effects on cell number (4) the possibility that dopamine neuron number is directly linked to cocaine-stimulated behavior and electrically-stimulated dopamine release and that these responses to cocaine are mediated through gonadal hormone modulation of midbrain dopamine neuron number. I discovered sex differences in midbrain dopamine neuron number; adult female rodents have more neurons in the SNpc and VTA. We also found that gonadectomy in adulthood reduced midbrain dopamine neuron number in females and increased neuron number in males, establishing the trophic effects of estrogen in the intact midbrain and possible suppressive effects of androgens. Treatment with agonists for estrogen receptor subtypes alpha and beta and androgen receptor reversed the effects of gonadectomy on cell number in females and males, respectively. In an effort to bridge cocaine-stimulated behavior and cell number in sham ovariectomized and ovariectomized females, we discovered cocaine-stimulated behavior, dopamine release and SNpc cell density were positively correlated in intact female rats, an effect that is lost with ovariectomy. This dissertation demonstrates that estrogen is critical for the maintenance of dopaminergic cell populations that enhance behavioral responses to psychostimulants in females, thereby contributing to the observed sex differences.</p> / Dissertation

An understanding of the damage evolution prior to crack initiation in advanced structural materials is of vital importance to the fatigue community in both academia and industry. Features known as the Persistent Slip Bands (PSBs) play an integral role in this damage evolution. Therefore, PSBs have been the focus of a lot of science-based investigations over the years. However, most existing studies in this area are restricted to analysis of PSBs in single crystal face centered cubic (FCC) materials. Moreover, these studies lack a quantitative analysis of the evolution of the fatigue damage (or PSBs) as a function of the material microstructure. This is especially true for relatively modern materials such as the Advanced High Strength Structural (AHSS) steels that are gaining a lot of importance in the automotive sector. Accordingly, the objective of this research is to quantitatively characterize evolution of PSBs in three AHSS steels having different microstructures as a function of number of fatigue cycles and strain amplitude. For this purpose strain controlled interrupted fatigue tests have been performed on two dual phase steels (DP-590 and DP-980) having different relative amounts of tempered martensite and a ferritic high strength low alloy steel (HR-590). Digital image analysis and Stereology have been used for unbiased quantitative characterization of the evolution of global geometry of the PSB colonies as function of number of fatigue cycles and strain amplitude. Evolution of PSB colonies has been couched in terms of variation of PSB colony volume fraction and total surface area unit volume, and total surface area of individual PSBs per unit volume and three-dimensional angular orientation distribution of the PSBs. For this purpose, new stereological techniques have been developed for estimation of the three-dimensional angular orientation distribution. The stereological data reveal that during strain controlled in these AHSS steels, volume fraction of the PSB colonies varies linearly with the their total surface area per unit volume. Detailed analysis of the stereological data leads to a simple geometric model for evolution of the PSB colonies in the three AHSS steels, which accounts for all observed data trends.

Stroke typically occurs in one hemisphere and often results in long-term disability in the contralateral body side (paretic side). Greater reliance on the non-paretic body side is used to compensate for this disability. Meanwhile, the brain undergoes degenerative and plastic changes in both hemispheres. Many previous studies have investigated post-stroke brain plasticity, and explored how it is shaped by behavioral experiences, to better understand the mechanisms of functional recovery. However, these studies have primarily focused on neurons and synapses. Given the abundant evidence that astrocytes actively control activity and plasticity of synapses, it seems reasonable to investigate how astrocytes are involved in behavior- and injury-driven brain plasticity. The central hypothesis of these studies is that synaptic plasticity underlying motor skill learning and post-stroke motor rehabilitation is coordinated with structural and functional plasticity of perisynaptic astrocytes. This was tested in a rat model of motor learning and "re-learning" after unilateral stroke-like damage to sensorimotor cortex. In the contralesional homotopic cortex, astrocytic volume varied with lesion size, as did the number of synapses. In the remaining motor cortex of the injured hemisphere, rehabilitative training with the paretic limb increased the proportion of astrocytic membrane apposed with synapses along with density of synapses. Furthermore, the percentage of synapses with astrocytic contacts was significantly correlated with functional outcome. Training with the non-paretic limb also induced greater synaptic density than controls in peri-infarct cortex, but functional outcome was negatively correlated with this and was not correlated with astrocytic contacts with synapses. These findings suggest that plasticity of, and association between, synapses and astrocytes vary with the type of experiences. Moreover, pharmacological upregulation of astrocytic glutamate uptake, which is one of the key ways that astrocytes modulate synaptic activity, interfered with functional recovery, supporting a critical role for astrocytic glutamate uptake in functional outcome following a stroke. Taken together, these studies contribute to better understanding of how lesions and experiences affect plasticity of astrocytes and synapses. These findings suggest that post-injury experiences alter astrocytic association with synapses, and that the coordinated plasticity of astrocytes and synapses is likely to be a critical mediator to functional outcome. / text

Delayed or impaired bone healing occurs in 5-10% of all fractures, yet cost effective solutions to enhance the healing process are limited. Lithium, a current treatment for bipolar disorder, is not clinically indicated for use in fracture management, but has been reported to positively influence bone biology. The objective of this study was to identify lithium administration parameters that maximize bone healing in a preclinical, rodent femur fracture model. Using a three factor, two level, design of experiments (DOE) approach, bone healing was assessed through mechanical testing and &mu;CT-image analysis. Significant improvements in healing were found at a low dose, later onset, longer duration treatment combination, with onset identified as the most influential parameter. The positive results from this DOE screening focuses the optimization phase towards further investigation of the onset component of treatment, and forms a crucial foundation for future studies evaluating the role of lithium in fracture healing.

Delayed or impaired bone healing occurs in 5-10% of all fractures, yet cost effective solutions to enhance the healing process are limited. Lithium, a current treatment for bipolar disorder, is not clinically indicated for use in fracture management, but has been reported to positively influence bone biology. The objective of this study was to identify lithium administration parameters that maximize bone healing in a preclinical, rodent femur fracture model. Using a three factor, two level, design of experiments (DOE) approach, bone healing was assessed through mechanical testing and &mu;CT-image analysis. Significant improvements in healing were found at a low dose, later onset, longer duration treatment combination, with onset identified as the most influential parameter. The positive results from this DOE screening focuses the optimization phase towards further investigation of the onset component of treatment, and forms a crucial foundation for future studies evaluating the role of lithium in fracture healing.

There is a need for simulation methodologies for multiphase three-dimensional microstructures that can be used in numerical simulations of material behavior or in exact computation of effective properties using microstructural correlation functions. Specifically, the methodology must be able to generate verifiably realistic microstructures, with complex morphology accurately represented. Striving to address that need, the research presented here develops a general microstructure simulation toolbox for multiphase two- and three-dimensional microstructures consisting of one connected phase and one or more particulate phases. Previous work by other researchers has found successful solutions to a variety of special cases of the general problem, but most of them are intended for binary microstructures, and nearly all simulate only two-dimensional microstructures. The toolbox presented here attempts to exceed those limitations. Its framework is a Metropolis stochastic-optimization routine running a simulated-anneal schedule, with particle position coordinates defining the configuration space and a range of forms available for the ﾓenergyﾔ? function. The toolbox allows several parameterizations of the microstructure, supplying all elementary properties (phase volume fractions, mean sizes, etc.) and some non-elementary properties (distributions of elementary properties, properties relating to inter-phase distances and morphology) of microstructures as possible parameters. The toolbox is able, as one special case, to simulate realistic microstructures of uniaxially compacted mixtures of elemental Al-Ti-B powders and achieve basic microstructure-processing correlation. Statistical tests involving microstructural correlation functions bear out the realism. The toolbox is also able to generate virtual microstructures for the same system, for use in the design of experiments (which are in fact high-strain-rate impact simulations), and for evaluating hypotheses involving achievable material properties. The Al-Ti-B powder compacts are potential advanced energetic materials that, when subjected to high-strain-rate impact (which may or may not constitute shock compression), explosively release heat by anaerobic reaction according as certain incompletely understood conditions are met or not. The study of those conditions and the mechanism of reaction initiation (carried out by a collaborator) is the specific application that the simulations in this work cater to. To ensure realistic morphology in simulated Al-Ti-B microstructures, this work included reconstruction (carried out by montage serial sectioning) of large three-dimensional volumes of Al-Ti and Al-B binary compacts for two sets of powders that yielded actual 3 D Ti and B particle images. Accordingly, advancement of the experimental technique of montage serial sectioning and a quantitative characterization of the real powder microstructures also formed part of this research. While only examples from Al-Ti-B powders are used throughout this work, it is clear that the methods will apply to other similar systems.

The concept of age has undergone a shift from a non-specific measure of chronological age, to an identification of underlying biological, psychological and functional factors leading to age-related changes over time. Loss of neurons (atrophy) and cognitive decline in healthy aging fit well in to this age paradigm. The aging brain is thought to undergo functional shifts in information processing in response to atrophy, which is conceptualised as a “Compensation Hypothesis” of cognitive aging. Using behavioural (reaction time, variability measures, and accuracy on the n-back task of working memory), structural (stereological cortical volume estimates) and functional (functional Magnetic Resonance Imaging) approaches, this study documents decreased whole brain, prefrontal and dorsolateral prefrontal cortex volumes in older individuals. Further, slower, less accurate, and more variable performance on the n-back task in older participants was accompanied by a posterior-to-anterior shift in processing, confirming the Compensation Hypothesis of cognitive aging. The behavioural data combined with structural and functional findings, suggest an aging brain that neuropsychologically compensates over time by paradoxically placing further processing demands on a structurally compromised dorsolateral prefrontal cortex. This produces adequate but slower, more variable, and less accurate performance compared to younger brains; compensation occurs in age, but is not complete. Decision making research has pointed to the important role of emotion in judgement, and has implicated the orbitofrontal cortex as critical for this processing modality. The structural data in this study showed preferentially less volume in the dorsolateral prefrontal cortex, but maintained cortical volume in the orbitofrontal cortex with age. Younger individuals took longer and maintained their accuracy with increasing complexity during the n-back task, with older participants decreasing their accuracy but not to the level of chance with increasing task complexity. As such, decision making on the n-back task may have shifted with age from the pure processing power of the structurally compromised dorsolateral prefrontal cortex to increasing reliance on emotionally-guided decision making inputs mediated by the intact orbitofrontal cortex resulting in adequate but not fully compensated performance in older people. These findings are discussed in relation to evolutionary pressures on the human working memory system, Hume’s concepts of reason and the passions, and to the emerging field of neuroeconomics. / Graduate