Thesis (Ph. D.)--Drexel University, 2005. / Includes abstract and vita. Includes bibliographical references (leaves 91-101).

Thesis (Ph.D.) - University of Queensland, 2005. / Includes bibliography.

Thesis (Ph. D.)--Drexel University, 2006. / Includes abstract and vita. Includes bibliographical references (leaves 118-128).

Dissertation (Ph.D.) -- University of Texas Southwestern Medical Center at Dallas, 2006. / Vita. Bibliography: pp. 193-208.

Structural changes in the frontal and temporal lobes and in subcortical white matter tracts often occur following closed head injury (CHI). In contrast to this well delineated structural pathology, the post-traumatic cognitively-related functional changes in these and other brain regions have not been adequately described. To characterize the long-term functional neuroanatomy of CHI, the present study compared regional cerebral blood flow (rCBF) patterns in 13 severely-injured, well-recovered, unmedicated patients to those from 13 well-matched healthy controls. rCBF was measured using oxygen-15 water intravenous bolus positron emission tomography (PET) while subjects performed the Wisconsin Card Sorting Test (WCST), an indicator of prefrontal lobe functioning that involves matching stimuli to a changing sorting principle based on external feedback, and a Cued Recall Memory Test (CRMT), which involves remembering semantically-related word pairs. The neuropsychological tasks were used to provoke specific neural systems believed to be important in task performance (the prefrontal cortex in the former, the hippocampus in the latter). Subjects also performed two specially designed sensorimotor control tasks to provide measures of baseline rCBF. Given the controversy regarding the statistical analysis of PET data, a two-pronged method was utilized: 1) Statistical Parametric Mapping, the state-of-the-art technique that examines rCBF throughout the entire brain, and 2) region of interest analysis, an anatomically-based method for examining rCBF in a limited set of brain regions. Between-group rCBF differences were tested in the four tasks separately and also in the two neuropsychological tasks after subtracting baseline rCBF (i.e., rCBF activation). To characterize the relationship between cerebral perfusion and behavior, correlations were performed between performance and rCBF activation (i.e., task-control) for each group separately, and between rCBF activation and an index of current neuropsychological functioning for the CHI patients. Analyses of each task separately revealed that, compared to controls, CHI patients showed lower rCBF in anterior cingulate cortex (ACC) and subcortical areas. Analyses of rCBF activation data revealed: 1) increases in left inferior frontal gyrus (including Broca's area) and left hippocampus of CHI patients relative to control subjects during the WCST, 2) a negative correlation between task performance and the right hippocampus during the WCST in CHI patients, and 3) correlations between the hippocampus and performance during the CRMT in the CHI patients that were in the opposite direction to those found in the control subjects. These neurofunctional changes are compatible with the structural and cognitive sequelae of CHI First, given a hypothesized role of the ACC in attentional processes, reduced rCBF in this region of CHI patients may relate to the persistent and often subtle difficulties in attention after CHI, whereas rCBF diminutions in subcortical regions may relate to diffuse damage to or deafferentation of subcortical regions in this CHI sample. Second, given similar (although slightly, but not significantly, poorer) performance on the WCST by the CHI patients, increased left prefrontal cortical activity may partially reflect behavioral compensation (e.g., subvocalization to aid memory during the task) and also physiological compensation for inefficiencies in other brain areas (e.g., subcortical regions). Finally, in light of the relatively poorer task performance of CHI patients (non-significant tendency in the WCST but highly significant in the CRMT), differences between the groups in the direction of the correlations between performance/cognition and hippocampal activation may suggest disorganization of hippocampal functioning in CHI patients. This exploratory and descriptive investigation identifies brain structures with post-traumatic changes that may be important to cognition. These results may provide evidence of both behavioral and neurophysiological compensation in patients with severe CHI. / Graduate

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CoordenaÃÃo de AperfeiÃoamento de Pessoal de NÃvel Superior / O lÃtex da planta medicinal Calotropis procera (Apocynaceae) à um produto quÃmico complexo, constituÃdo de uma diversidade de molÃculas e tem sido amplamente utilizado na medicina popular sobre enfermidades dermatolÃgicas. O potencial da fraÃÃo de proteÃnas do lÃtex (PL) em induzir danos celulares foi avaliado pelos testes de MTT e LDH. AlÃm disso, PL foi utilizada no preparo de uma biomembrana associada ao poli (Ãlcool vinÃlico) (BioMem PVA/PL 0,2% e 1%). As propriedades fÃsico-quÃmica das membranas controle (PVA 1%) e teste (BioMem PVA/PL 1%) foram avaliadas atravÃs da espectroscopia de absorÃÃo na regiÃo do infravermelho com transformada de Fourier (FTIR) e anÃlise termogravimÃtrica (TGA). O efeito das proteÃnas do lÃtex na cicatrizaÃÃo foi investigado apÃs induÃÃo de feridas incisionais ou excisionais no dorso de camundongos, seguido da implantaÃÃo da BioMem PVA/PL 0,2% ou 1%. O processo cicatricial foi avaliado de acordo com os parÃmetros: anÃlise macroscÃpica (induÃÃo da neoformaÃÃo tecidual em feridas incisionais e edema, hiperemia, reduÃÃo da Ãrea e reepitelizaÃÃo em feridas excisionais); microscÃpica (degranulaÃÃo de mastÃcitos, edema, infiltrado de leucÃcitos, nÃmero de fibroblastos e colagÃnese). Marcadores e mediadores da fase inflamatÃria (MPO, nitrito, IL-1β e TNF-α) foram avaliados em feridas incisionais e excisionais. O efeito da BioMem PVA/PL sobre a densidade microvascular foi avaliado em feridas incisionais. O possÃvel efeito de PL em estimular diretamente macrÃfagos foi investigado em uma cultura de cÃlulas. PL nÃo apresentou in vitro citotoxicidade sobre neutrÃfilos humanos, visto pela baixa atividade de LDH e a alta viabilidade dos neutrÃfilos pelo teste do MTT. A anÃlises fÃsico-quÃmicas demonstraram que nÃo ocorreram interaÃÃes fortes do PVA com PL, jà que nÃo foi observada a formaÃÃo de novas bandas ou o deslocamento destas. AlÃm disso, a solubilizaÃÃo destes compostos tornou a BioMem PVA/PL termicamente mais estÃvel. Nos modelos experimentais de cicatrizaÃÃo, as anÃlises macroscÃpicas demonstraram que BioMem PVA/PL 0,2% e 1% induziram a neoformaÃÃo tecidual em feridas incisionais nos dias 2, 7 e 14 apÃs a implantaÃÃo das membranas. Entretanto, nÃo interferiu na densidade da microvascularizaÃÃo ao tecido neoformado. Em feridas excisionais, BioMem PVA/PL 0,2% induziu aumento do edema, mas nÃo interferiu na hiperemia na fase inflamatÃria. Na fase proliferativa da cicatrizaÃÃo, acelerou a reduÃÃo da Ãrea das feridas e favoreceu a reepitelizaÃÃo. A anÃlise microscÃpica de feridas incisionais e excisionais evidenciou que BioMem PVA/PL 0,2% ou 1% conferiu um estÃmulo à degranulaÃÃo de mastÃcitos, edema e migraÃÃo de leucÃcitos na fase inflamatÃria do processo cicatricial. Na fase proliferativa foi evidenciado aumento na populaÃÃo de fibroblastos e colagÃnese. A anÃlise da reepitelizaÃÃo de feridas excisionais evidenciou um aumento da espessura do epitÃlio neoformado. A implantaÃÃo da BioMem PVA/PL 0,2% aumentou os nÃveis de marcadores e mediadores da resposta inflamatÃria, tais como MPO, nitrito, IL-1β e TNF-α. MacrÃfagos cultivados de camundongos e estimulados com PL foram induzidos a liberaÃÃo de TNF- e IL-1β. A anÃlise integrada de todos os resultados sugerem que as proteÃnas do lÃtex (PL) atuam significativamente na fase inflamatÃria da cicatrizaÃÃo, o que parece influenciar diretamente as fases subseqÃentes do processo cicatricial. / The latex of the medicinal plant Calotropis procera (Apocynaceae) is a complex chemical, consisting of a variety of molecules and has been widely used in folk medicine on skin diseases. The potential of the latex proteins fraction (LP) in induce cell damage was assessed by MTT and LDH tests. Moreover, LP was used to prepare a biomembrana associated with poly (vinyl alcohol) (BioMem PVA/PL 0,2% and 1%).The physical-chemical properties of control (PVA 1%) and test (BioMem PVA/PL 1%) were assessed by infrared spectroscopy (FTIR) and thermogravimetric (TGA) analysis. The effect of laticifers proteins at healing was investigated after induction of incision or excision wounds on the back of mices, followed by implantation of BioMem PVA/PL 0,2% or 1%. Healing process was evaluated by the parameters: macroscopic analyses (induction of tissue neoformation of incisional wounds and edema, hyperemia, area reduction and re-epithelialization of excision wounds); microscopic (mast cell degranulation, edema, leucocyte infiltrate, number of fibroblasts and collagenesis). Inflammatory markers and mediators (MPO, nitrite, IL-1β e TNF-α) were evaluated in incision and excision wounds. Microvessel density was evaluated in incision wounds. The possible effect of soluble protein fraction (LP) to directly stimulate macrophages was investigated in a cell culture. PL shows no cytotoxicity in vitro in human neutrophils, since the low activity of LDH and high vibility of neutrophils by MTT test. The physico-chemical analysis showed that no strong interaction with PVA and LP occurred, since was not observed the formation of new bands or the displacement of these. Futhermore, the solubility of these compounds became the BioMem PVA/PL more thermally stable. At experimental healing models, macroscopic analyses showed that BioMem PVA/PL 0,2% e 1% leads tissue neoformation in incisional wounds at the 2, 7 and 14 days after membrane implantation. However, no effect on the microvascular density to neoformed tisse of incisional wounds were seen. At excisional wounds, BioMem PVA/PL 0,2% induce increase of edema, but not hyperemia at inflammatory phase. Moreover, accelerated the reduction in the wound area and an improved re-epithelialization at proliferative phase of wound healing. Microscopic analysis of incisional and excisional wounds showed that BioMem PVA/PL 0,2% or 1% lead a stimulus to mast cell degranulation, edema, leucocyte migration at inflammatory phase of the cicatricial process. At the proliferative phase, was evidenced an increase in the population of fibroblasts and collagenesis. Re-epithelialization of excisional wounds showed an increased thickness of the newly formed epithelium. BioMem PVA/PL 0,2% implantation increased the levels of markers and mediators of the inflammatory response, such as MPO, nitrite, IL-1β and TNF-α. Culture mouse macrophages stimulated with PL were induced to release of TNF- e IL-1β. Integrated analysis of all results suggest that PL act significantly in the inflammatory phase of healing, which seems to directly influence the subsequent phases of the healing process.

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