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Angiogeneze v okrajových částech mozkové ischemie / Angiogenesis in cerebral ischemia border zoneNulíčková, Radka January 2011 (has links)
Title of work: Angiogenesis in the border zones of cerebral ischemia The aims of work: The aim of this diploma thesis was to determinate the parameter of the angiogenesis after focal brain ischemia induced by photothrombosis in P7 rat pups. This thesis should answer the question, if there is some difference in vascular density according to the distance from the lesion and if there is some diference between the vascular density in normal brain and the brain with the lesion. Materials and Methods: Cerebral ischemia was induced in P7 rat pups with photothombic method. Intravenous administration of bengal rose dye was followed by illumination of beam of green laser over the cortex for 5 minutes at 0,5s on/off cycles. Bengal rose dye is potent photosensitive substance and the ilumination resulted in agregation of trombocytes and thrombosis. Cerebral ischemia evolved due to the thrombosis. After two months, the animals were overdosed with urethane and transcardially perfused. Then thein brain were cryoprotected and then cut into 50µm slices and prepaired for histological staining. The stained slices were the investigated under mikroskope. The microvessels in the cortex were counted and the density was estimated by stereological method. Results: Two brain with ischemic lesion and two control brain were...
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eCG e densidade vascular em úteros bovinos / eCG and vascular density of bovine uterusPinto, Joana Mona e 18 December 2009 (has links)
A dinâmica folicular, bem como a preparação do endométrio para a gestação, é dependente do estabelecimento de vascularização adequada. Alguns estudos apontam para a importância do tratamento com eCG após o emprego de protocolos de IATF, de forma a aumentar as taxas de ovulação e prenhez, e para a uma possível propriedade angiogênica, influenciando o fluxo sanguíneo e a vascularização. O objetivo do presente trabalho foi avaliar a influência do eCG na espessura e área do endométrio e miométrio, calibre e área da artéria uterina do corno ipsilateral e também na densidade vascular do endométrio e miométrio de vacas submetidas a tratamentos de sincronização (grupo controle), estimulação do folículo dominante e superovulação. Para tal, foram utilizadas 16 vacas mestiças de Nelore ciclando com escore corporal entre 2 e 3 que foram divididas em 3 grupos de acordo com tratamento hormonal: o grupo controle foi submetido apenas ao protocolo de sincronização da ovulação (n=5), o grupo estimulado recebeu 400 UI de eCG no dia 4 após início do protocolo (n=6) e o grupo superovulado recebeu 2000UI de eCG no dia 8 após início do tratamento (n=5). No dia 5 após a ovulação (p.o) foram realizados exames ultrasonográficos para mensuração do miométrio e endométrio dos cornos uterinos, bem como para análise do calibre da artéria uterina. Os animais foram abatidos, os úteros coletados no dia 6 p.o. e imediatamente fixados em paraformaldeido a 4%. Para análise da densidade vascular, foi realizada imunohistoquímica para o KDR com o intuito de marcação das células endoteliais e contagem dos vasos por estereologia. As espessuras e áreas, tanto do miométrio quanto do endométrio, não foram influenciadas pelos tratamentos. A artéria uterina nos animais superovulados apresentou maior calibre do que nos animais do grupo controle (p< 0,05). A densidade vascular do endométrio nos animais estimulados foi menor quando comparada à dos animais do grupo controle (p<0,05), e no grupo superovulado, a densidade vascular apresentou-se diminuída (p<0,05) no miométrio. Os resultados obtidos sugerem que o aumento de vascularização do endométrio após tratamentos com ECG, relatado em outros trabalhos, seja provavelmente influenciado pelo concepto e não exclusivamente pelo tratamento. Além disso, a influência do eCG no útero pode ser dependente tanto da dose quanto do estágio do ciclo estral. / Follicle dynamic, as well as the uterus preparation for gestation, is dependent of an adequate vascularization establishment. Currently, many studies point towards the importance of eCG treatment following fixed-time artificial insemination (FTAI), in a manner to improve ovulation and gestational rates. Moreover, eCG have been implicated in angiogenesis, leading to important changes in uterine blood flow and vascularization. Thus, the present study was designed to investigate the influence of eCG on the endometrial and myometrial thickness and area; on the caliber and area of the uterine artery of the ipsilateral horn; and, on endometrial and myometrial uterine vascular density of cows submitted to sincronization (control group), stimulation (stimulated group), and superovulatory (superovulated group) treatments. For that, we used 16 cows with body score between 2 and 3, randomly distributed into the 3 above described groups: the cows of the control group (n = 5) did not receive eCG, while the cows of the stimulated group (n = 6) and superovulated group 3 (n = 5) received, respectively, 400 UI and 2000 UI of eCG at days 4 and 8 after the protocol beginning. For endometrial and myometrial measurements, as well as for uterine artery caliber and area analysis, ultrasonographic evaluations were done at day 5 after ovulation. At day 6, the animals were slaughtered, the uterus were harvested and fixed in 4% paraformaldehyde. In order to analyze the vascular density, immunohistochemistry for KDR detection and blood vessels counting by stereology were performed. In all studied groups, treatments did not influence either uterine wall thickness or area. In the superovulated animals, the uterine artery presented higher caliber than in the control ones (P<0.05). Endometrial vascular density of stimulated animals in the ipsilateral uterine horn was lower when compared to that of the control (P<0.05),and in the superovulated, vascular density was lower in the myometrium (P<0.05). These results suggest that the endometrial vascularization increase following eCG treatment, described by other groups, is probably influenced by the concept, not only by the treatment. Moreover, the influence of eCG in the uterus could be dependent of both the hormonal dosis and the estrous cycle phase.
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eCG e densidade vascular em úteros bovinos / eCG and vascular density of bovine uterusJoana Mona e Pinto 18 December 2009 (has links)
A dinâmica folicular, bem como a preparação do endométrio para a gestação, é dependente do estabelecimento de vascularização adequada. Alguns estudos apontam para a importância do tratamento com eCG após o emprego de protocolos de IATF, de forma a aumentar as taxas de ovulação e prenhez, e para a uma possível propriedade angiogênica, influenciando o fluxo sanguíneo e a vascularização. O objetivo do presente trabalho foi avaliar a influência do eCG na espessura e área do endométrio e miométrio, calibre e área da artéria uterina do corno ipsilateral e também na densidade vascular do endométrio e miométrio de vacas submetidas a tratamentos de sincronização (grupo controle), estimulação do folículo dominante e superovulação. Para tal, foram utilizadas 16 vacas mestiças de Nelore ciclando com escore corporal entre 2 e 3 que foram divididas em 3 grupos de acordo com tratamento hormonal: o grupo controle foi submetido apenas ao protocolo de sincronização da ovulação (n=5), o grupo estimulado recebeu 400 UI de eCG no dia 4 após início do protocolo (n=6) e o grupo superovulado recebeu 2000UI de eCG no dia 8 após início do tratamento (n=5). No dia 5 após a ovulação (p.o) foram realizados exames ultrasonográficos para mensuração do miométrio e endométrio dos cornos uterinos, bem como para análise do calibre da artéria uterina. Os animais foram abatidos, os úteros coletados no dia 6 p.o. e imediatamente fixados em paraformaldeido a 4%. Para análise da densidade vascular, foi realizada imunohistoquímica para o KDR com o intuito de marcação das células endoteliais e contagem dos vasos por estereologia. As espessuras e áreas, tanto do miométrio quanto do endométrio, não foram influenciadas pelos tratamentos. A artéria uterina nos animais superovulados apresentou maior calibre do que nos animais do grupo controle (p< 0,05). A densidade vascular do endométrio nos animais estimulados foi menor quando comparada à dos animais do grupo controle (p<0,05), e no grupo superovulado, a densidade vascular apresentou-se diminuída (p<0,05) no miométrio. Os resultados obtidos sugerem que o aumento de vascularização do endométrio após tratamentos com ECG, relatado em outros trabalhos, seja provavelmente influenciado pelo concepto e não exclusivamente pelo tratamento. Além disso, a influência do eCG no útero pode ser dependente tanto da dose quanto do estágio do ciclo estral. / Follicle dynamic, as well as the uterus preparation for gestation, is dependent of an adequate vascularization establishment. Currently, many studies point towards the importance of eCG treatment following fixed-time artificial insemination (FTAI), in a manner to improve ovulation and gestational rates. Moreover, eCG have been implicated in angiogenesis, leading to important changes in uterine blood flow and vascularization. Thus, the present study was designed to investigate the influence of eCG on the endometrial and myometrial thickness and area; on the caliber and area of the uterine artery of the ipsilateral horn; and, on endometrial and myometrial uterine vascular density of cows submitted to sincronization (control group), stimulation (stimulated group), and superovulatory (superovulated group) treatments. For that, we used 16 cows with body score between 2 and 3, randomly distributed into the 3 above described groups: the cows of the control group (n = 5) did not receive eCG, while the cows of the stimulated group (n = 6) and superovulated group 3 (n = 5) received, respectively, 400 UI and 2000 UI of eCG at days 4 and 8 after the protocol beginning. For endometrial and myometrial measurements, as well as for uterine artery caliber and area analysis, ultrasonographic evaluations were done at day 5 after ovulation. At day 6, the animals were slaughtered, the uterus were harvested and fixed in 4% paraformaldehyde. In order to analyze the vascular density, immunohistochemistry for KDR detection and blood vessels counting by stereology were performed. In all studied groups, treatments did not influence either uterine wall thickness or area. In the superovulated animals, the uterine artery presented higher caliber than in the control ones (P<0.05). Endometrial vascular density of stimulated animals in the ipsilateral uterine horn was lower when compared to that of the control (P<0.05),and in the superovulated, vascular density was lower in the myometrium (P<0.05). These results suggest that the endometrial vascularization increase following eCG treatment, described by other groups, is probably influenced by the concept, not only by the treatment. Moreover, the influence of eCG in the uterus could be dependent of both the hormonal dosis and the estrous cycle phase.
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Experimental Studies on the Vasculature of Endogenous and Transplanted Islets of LangerhansMattsson, Göran January 2003 (has links)
<p>The blood vessels of the pancreatic islets are of crucial importance for oxygen and metabolite supply as well as dispersal of secreted hormones. In addition to this, endothelial cells have an important role in the revascularization process after islet transplantation. Previous studies have reported signs of poor engraftment of transplanted islets, presumably due to impaired revascularization. The aims of this thesis were to investigate the revascularization process of transplanted islets and to examine the role of islet endothelial cells. In this context, the lectin Bandeiraea simplicifolia was found to stain endothelium of both endogenous and transplanted pancreatic islets. By using this lectin we investigated the vascular density of both endogenous and islets transplanted syngeneically beneath the renal capsule, into the spleen or intraportally into the liver of normoglycemic C57BL/6 mice. One month post-transplantation, a time point when the grafts are assumed to be completely revascularized, the vascular density was decreased at all three implantation sites when compared to endogenous islets. Furthermore, most of the blood vessels were located in the graft connective tissue stroma. Similar results were obtained when islet transplant vascular density was determined six months post-transplantation and in cured diabetic animals after one month. In order to evaluate the function of intraportally transplanted islets, we developed a method to retrieve such islets. We treated the implantation organ (liver) first enzymatically (collagenase) and then mechanically, thereafter we could re-isolate the transplanted islets for further in vitro studies. The retrieved islets had a decreased insulin relase, insulin content and glucose oxidation rate when compared to non-transplanted control islets. To understand the role of islet endothelium in the revascularization of transplanted islets we performed angiogenesis GEArray studies on islet endothelial cells, from non-cultured, cultured and transplanted islets. We found that the islet endothelium expressed mRNA for both inhibitors and inducers of angiogenesis, and that this expression differed with time. The functional consequences of this remain to be determined. In summary, the results presented above provide a useful platform for future studies of the morphology and function of islet endothelial cells, especially with a view for elucidating changes induced by islet transplantation.</p>
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Implantation-Site Dependent Differences in Engraftment and Function of Transplanted Pancreatic IsletsLau, Joey January 2008 (has links)
<p>Transplanting pancreatic islets into the liver through the portal vein is currently the most common procedure in clinical islet transplantations for treating patients with brittle type 1 diabetes. However, most islet grafts fail within a 5-year period necessitating retransplantation. The vascular connections are disrupted at islet isolation and implanted islets depend on diffusion of oxygen and nutrients in the immediate posttransplantation period. Rapid and efficient revascularization is of utmost importance for the survival and long-term function of transplanted islets. </p><p>In this thesis, the influence of the implantation microenvironment for islet engraftment and function was studied. Islets were transplanted into the liver, the renal subcapsular site or the pancreas. Islets implanted into the liver contained fewer glucagon-positive cells than islets implanted to the kidney and endogenous islets. Intraportally transplanted islets responded with insulin and glucagon release to secretagogues, but only when stimulated through the hepatic artery. Thus, the intrahepatic grafts were selectively revascularized from the hepatic artery. The vascular density in human islets transplanted into the liver of athymic mice was markedly lower when compared to human islets grafted to the kidney. Islets implanted into their physiological environment, the pancreas, were markedly better revascularized. Insulin content, glucose-stimulated insulin release, (pro)insulin biosynthesis and glucose oxidation rate were markedly decreased in transplanted islets retrieved from the liver, both when compared to endogenous and transplanted islets retrieved from the pancreas. Only minor changes in metabolic functions were observed in islets implanted into the pancreas when compared to endogenous islets. </p><p>The present findings demonstrate that the microenvironment has a major impact on the engraftment of transplanted islets. Elucidating the beneficial factors that promote engraftment would improve the survival and long-term function of transplanted islets. Ultimately, islet transplantation may be provided to an increased number of patients with type 1 diabetes.</p>
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Experimental Studies on the Vasculature of Endogenous and Transplanted Islets of LangerhansMattsson, Göran January 2003 (has links)
The blood vessels of the pancreatic islets are of crucial importance for oxygen and metabolite supply as well as dispersal of secreted hormones. In addition to this, endothelial cells have an important role in the revascularization process after islet transplantation. Previous studies have reported signs of poor engraftment of transplanted islets, presumably due to impaired revascularization. The aims of this thesis were to investigate the revascularization process of transplanted islets and to examine the role of islet endothelial cells. In this context, the lectin Bandeiraea simplicifolia was found to stain endothelium of both endogenous and transplanted pancreatic islets. By using this lectin we investigated the vascular density of both endogenous and islets transplanted syngeneically beneath the renal capsule, into the spleen or intraportally into the liver of normoglycemic C57BL/6 mice. One month post-transplantation, a time point when the grafts are assumed to be completely revascularized, the vascular density was decreased at all three implantation sites when compared to endogenous islets. Furthermore, most of the blood vessels were located in the graft connective tissue stroma. Similar results were obtained when islet transplant vascular density was determined six months post-transplantation and in cured diabetic animals after one month. In order to evaluate the function of intraportally transplanted islets, we developed a method to retrieve such islets. We treated the implantation organ (liver) first enzymatically (collagenase) and then mechanically, thereafter we could re-isolate the transplanted islets for further in vitro studies. The retrieved islets had a decreased insulin relase, insulin content and glucose oxidation rate when compared to non-transplanted control islets. To understand the role of islet endothelium in the revascularization of transplanted islets we performed angiogenesis GEArray studies on islet endothelial cells, from non-cultured, cultured and transplanted islets. We found that the islet endothelium expressed mRNA for both inhibitors and inducers of angiogenesis, and that this expression differed with time. The functional consequences of this remain to be determined. In summary, the results presented above provide a useful platform for future studies of the morphology and function of islet endothelial cells, especially with a view for elucidating changes induced by islet transplantation.
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Implantation-Site Dependent Differences in Engraftment and Function of Transplanted Pancreatic IsletsLau, Joey January 2008 (has links)
Transplanting pancreatic islets into the liver through the portal vein is currently the most common procedure in clinical islet transplantations for treating patients with brittle type 1 diabetes. However, most islet grafts fail within a 5-year period necessitating retransplantation. The vascular connections are disrupted at islet isolation and implanted islets depend on diffusion of oxygen and nutrients in the immediate posttransplantation period. Rapid and efficient revascularization is of utmost importance for the survival and long-term function of transplanted islets. In this thesis, the influence of the implantation microenvironment for islet engraftment and function was studied. Islets were transplanted into the liver, the renal subcapsular site or the pancreas. Islets implanted into the liver contained fewer glucagon-positive cells than islets implanted to the kidney and endogenous islets. Intraportally transplanted islets responded with insulin and glucagon release to secretagogues, but only when stimulated through the hepatic artery. Thus, the intrahepatic grafts were selectively revascularized from the hepatic artery. The vascular density in human islets transplanted into the liver of athymic mice was markedly lower when compared to human islets grafted to the kidney. Islets implanted into their physiological environment, the pancreas, were markedly better revascularized. Insulin content, glucose-stimulated insulin release, (pro)insulin biosynthesis and glucose oxidation rate were markedly decreased in transplanted islets retrieved from the liver, both when compared to endogenous and transplanted islets retrieved from the pancreas. Only minor changes in metabolic functions were observed in islets implanted into the pancreas when compared to endogenous islets. The present findings demonstrate that the microenvironment has a major impact on the engraftment of transplanted islets. Elucidating the beneficial factors that promote engraftment would improve the survival and long-term function of transplanted islets. Ultimately, islet transplantation may be provided to an increased number of patients with type 1 diabetes.
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A role for toll-like receptor-4 in pulmonary angiogenesis following multiple exposures to swine barn airJuneau, Vanessa Jade 14 June 2007
Swine barn air is a heterogeneous mixture of dust, bacteria and irritant chemicals including ammonia and hydrogen sulphide. Gram-negative bacteria are commonly found in swine barn air and significantly contribute to pulmonary disease in unprotected swine barn workers, through the endotoxin moiety, lipopolysaccharide (LPS). Toll-like Receptor-4 is the ligand for LPS. It is found on many cell types including monocytes, macrophages, neutrophils, endothelial cells, and to a lesser extent, epithelial cells. The severity and outcome of acute lung injury following barn air exposures depends upon the balance between epithelial and vascular endothelial repair mechanisms, including angiogenesis. Vascular Endothelial Growth Factor (VEGF) is an endothelial mitogen produced by mesenchymal and alveolar Type II epithelial cells and by activated bronchial airway epithelial cells. Research investigating the role of cytokines in angiogenesis has shown that close proximity of immune cells and endothelial cells modulates the production of various compounds that regulate vascular function. Given that LPS is the ligand for TLR4 there appeared to be a role for TLR4 in angiogenesis, particularly following endotoxin exposure. To determine whether this was occurring, we examined whether exposure to swine barn air alters vascular density in the lungs and the role of TLR4 using a murine model. Toll-like Receptor-4 wild-type (C3HeB/FeJ) and TLR4 mutant (C3H/HeJ) mice were obtained and exposed to swine barn air for 1-, 5-, or 20-days for 8 hours/day. Wild-type animals showed a 127% increase in vascular density after 20-days barn air exposure. Vascular Endothelial Growth Factor-A protein levels were decreased by 0.62-fold after one-day swine barn air exposure in wild-type animals, indicating that VEGF-A is being used as a pro-angiogenic mitogen. Transcription of VEGF-A mRNA was increased in wild-type animals after all swine barn air exposure periods. The receptor VEGFR-1 showed increased mRNA transcription over all time points. These effects were only observed in TLR4 wild-type animals, indicating that these effects are mediated by TLR4. Further, VEGF-A and VEGFR-1 appear to be involved in the manifestation of TLR4-induced angiogenesis in the lung.
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A role for toll-like receptor-4 in pulmonary angiogenesis following multiple exposures to swine barn airJuneau, Vanessa Jade 14 June 2007 (has links)
Swine barn air is a heterogeneous mixture of dust, bacteria and irritant chemicals including ammonia and hydrogen sulphide. Gram-negative bacteria are commonly found in swine barn air and significantly contribute to pulmonary disease in unprotected swine barn workers, through the endotoxin moiety, lipopolysaccharide (LPS). Toll-like Receptor-4 is the ligand for LPS. It is found on many cell types including monocytes, macrophages, neutrophils, endothelial cells, and to a lesser extent, epithelial cells. The severity and outcome of acute lung injury following barn air exposures depends upon the balance between epithelial and vascular endothelial repair mechanisms, including angiogenesis. Vascular Endothelial Growth Factor (VEGF) is an endothelial mitogen produced by mesenchymal and alveolar Type II epithelial cells and by activated bronchial airway epithelial cells. Research investigating the role of cytokines in angiogenesis has shown that close proximity of immune cells and endothelial cells modulates the production of various compounds that regulate vascular function. Given that LPS is the ligand for TLR4 there appeared to be a role for TLR4 in angiogenesis, particularly following endotoxin exposure. To determine whether this was occurring, we examined whether exposure to swine barn air alters vascular density in the lungs and the role of TLR4 using a murine model. Toll-like Receptor-4 wild-type (C3HeB/FeJ) and TLR4 mutant (C3H/HeJ) mice were obtained and exposed to swine barn air for 1-, 5-, or 20-days for 8 hours/day. Wild-type animals showed a 127% increase in vascular density after 20-days barn air exposure. Vascular Endothelial Growth Factor-A protein levels were decreased by 0.62-fold after one-day swine barn air exposure in wild-type animals, indicating that VEGF-A is being used as a pro-angiogenic mitogen. Transcription of VEGF-A mRNA was increased in wild-type animals after all swine barn air exposure periods. The receptor VEGFR-1 showed increased mRNA transcription over all time points. These effects were only observed in TLR4 wild-type animals, indicating that these effects are mediated by TLR4. Further, VEGF-A and VEGFR-1 appear to be involved in the manifestation of TLR4-induced angiogenesis in the lung.
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Identificação da expressão do vascular endothelial growth factor (VEGF) pela contagem de células marcadas imunoistoquimicamente no omento de ratos após ligadura arterial e após ligadura venosaZart, Ronald Paulo Pinto January 2007 (has links)
O sistema cardiovascular está estrutural e funcionalmente disposto de modo “circular”. Situações de obstrução do fluxo sanguíneo determinam o aparecimento de mecanismos que visam suplantar tais interrupções e manter a circularidade íntegra. À nível molecular, o principal elemento envolvido nestes mecanismos é o Vascular Endothelial Growth Factor (VEGF). A expressão do VEGF quando há oclusão arterial está bem documentada, faltando elementos com relação à oclusão venosa. Neste estudo objetivamos verificar se a oclusão à nível arterial determina uma expressão do VEGF diferente daquela que ocorre se a oclusão acontecer à nível venoso. Para isso randomizamos dois grupos de ratos de experimentação. Em um grupo realizamos a oclusão da aorta infra-renal e em outro a oclusão da veia cava infra-renal. Posteriormente medimos a expressão do VEGF através da contagem do número de células marcadas imunoistoquimicamente no omento destes ratos. O resultado demonstrou que a expressão do VEGF, quando analisada pelo método proposto foi igual no grupo da oclusão venosa e no grupo da oclusão arterial. / The cardiovascular system is structurally and functionally circular. Situations in which there is obstruction to blood flow trigger mechanisms to bypass these blockages and maintenance the integrity of the circularity. At the molecular level the main factor involved is the Vascular Endothelial Growth Factor (VEGF). The VEGF expression associated with arterial occlusion is well documented but is lacking evidence when venous occlusion occurs. This study aimed to verify if the expression of VEGF when an occlusion occurs at venous level is the same or different from that caused at the arterial level. Two groups of rats were randomized by infra-renal aortic occlusion or inferior vena cava occlusion. VEGF was measured by counting the immunohistochemistry method marked cells at the omentum level. It was demonstrated that the VEGF expression is the same in the venous group obstruction as the arterial obstruction group.
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