Procurou-se confirmar a existência de alterações estruturais em corpos carotídeos de ratos expostos à hiperóxia que pudessem explicar a atenuação da resposta fisiológica à hipóxia nessas condições descrita na literatura. Também testamos a hipótese de haver um desvio de fluxo sanguíneo para os capilares intraglômicos em situações de hiperoxigenação hiperbárica.15 ratos machos Wistar adultos foram divididos em 3 grupos e expostos a 2,4 ATA por 6 horas, a 3,0 ATA por 6 horas e a ar ambiente (grupo controle). Os resultados obtidos através de análise histológica e morfométrica mostraram: a) nenhuma alteração da arquitetura dos corpos carotídeos, mas as células expostas à dose mais elevada apresentaram-se com citoplasma desarranjado, confirmado pela microscopia eletrônica; b) um aumento significativo da densidade volumétrica de capilares preenchidos por hemácias, mas não do estroma intersticial, no grupo exposto à dose mais elevada de O2 c) uma vasoconstricção significativa das arteríolas maiores em todas as doses de oxigênio empregadas no estudo e das arteríolas menores na dose mais elevada de O2; d) variações significativas na proporção das variantes de células glômicas no grupo exposto a menor dose de O2; e) mitocôndrias com poucas cristas, tanto nas células glômicas quanto nas terminações nervosas, embora nas primeiras apresentem-se bem deformadas; f) proliferação membranosa citoplasmática com aumento de REG e Golgi nas células glômicas e sustentaculares. Esses resultados sugerem um desvio do fluxo dos vasos mais calibrosos em direção aos capilares intraglômicos, confirmando nossa hipótese inicial e indicam que o oxigênio, dependendo da dose utilizada, exerce um efeito tóxico importante sobre os corpos carotídeos, com alterações significativas da ultraestrutura das células glômicas e terminações nervosas. / We sough to confirm the existence of structural alterations in rat carotid bodies exposed to hyperoxia that could explain the attenuation of the ventilatory hypoxic drive (HD) by hyperoxic conditions described in the literature. We also tested the hypothesis of there being a deviation of blood flow toward intraglomic capillaries in situations of hyperbaric oxygenation (HBO).15 adult male Wistar rats were divided in 3 groups and exposed to O2 at 2.4 ATA for 6 hours, at 3.0 ATA for 6 hours and to air at 1.0 ATA (control group). The results obtained through histological and morphometric analysis showed: a) no alteration in the architecture of the carotid bodies, but the cytoplasm of the cells exposed to the highest dose were disarranged, a feature confirmed by electron microscopy; b) a significant increase in volume density of capillaries filled out by red blood cells but not of interstitial stroma in the group exposed to O2 at the highest dose; c) a significant vasoconstriction of larger arterioles in all doses of oxygen employed in the study and of smaller arterioles at the highest dose of O2; d) significant variations in the proportion of glomic cell variants in the group exposed to the lowest dose of O2; e) mitochondria with few cristae, so in glomic cells as in nerve-endings, although in the former they were very deformed; f) cytoplasmic membranous proliferation with an increase of endoplasmic reticulum and Golgi apparatus in glomic and sustentacular cells. These results suggest a deviation of blood flow from more calibrated vessel toward intraglomic capillaries, confirming our initial hypothesis and indicate that oxygen, depending on the dose used, exerts an important toxic effect on rat carotid body with significant alterations of glomic cell and nerve-endings ultrastructure.

Introduction : L’arrêt cardiaque réfractaire est défini par l’absence du retour à l’activité circulatoire spontané (RACS) après 30 minutes de réanimation cardiopulmonaire médicalisé. ExtraCorporeal Membrane Oxygenation (ECMO) représente une thérapie alternative urgente dans cette population. L’hémodynamique post la réanimation cardiopulmonaire extracorporel (E-CRP) est un entité complexe et le pris en charge dans les premières heures suivant l’implantation d’ECMO n’est pas bien décrit. L’objectif de cette étude est d’évaluer l’effet de deux stratégie de débit d’ECMO dans un modèle porcin d’arrêt cardiaque réfractaire sur les conséquences métaboliques, microcirculatoires et inflammatoires.Matériels et Méthodes : l’arrêt cardiaque a été induit par la ligature l’artère intraventriculaire antérieure (IVA) chez 18 cochons. E-RCP a été initié après 40 minutes de low-flow avec un débit d’ECMO bas de 30-35 ml.kg-1.min-1 ou un débit d’ECMO standard de 65-70 ml.kg-1.min-1, avec la même pression artérielle moyenne (PAM) au niveau de 65 mmHg. Les paramètres hémodynamiques et métaboliques ont été évalués avec la clairance de lactate et le débit sanguin carotidien. Les paramètres microcirculatoires ont été évalués par la microcirculation sublinguale avec l’imagerie de SDF et NIRS. Cytokines inflammatoires ont été mesurés avec un plateforme de ELISA multiplexe. Résultats : Pas de différence entre les deux groups à H basale et à l’initiation d’ECMO (H0). La clairance de lactate était plus faible dans le groupe débit bas comparé au groupe débit standard (6.67[-10.43-18.78] vs. 47.41[19.54, 70.69] %, p=0.04). Le débit carotidien était plus bas significativement (p<0.005) dans le groupe débit bas pendant les dernières quatre heures malgré le même niveau de la pression artérielle moyenne. Pour les paramètres microcirculatoires, le flux microcirculatoire sublingual évalué par SDF et le StO2 par NIRS ont été altéré transitoirement à H3 dans le groupe débit bas. Le niveau de cytokine IL-6 était plus élevé significativement dans le groupe débit bas à la fin d’expérimentation. Conclusions : Une réanimation à objectif de débit d’ECMO bas 35 ml.kg-1.min-1 versus standard 70ml.kg-1.min-1dans les six premières heures d’un ACR réfractaire n’est pas associé à une meilleure réversion des conséquences métaboliques, microcirculatoire et inflammatoire avec un objectif de PAM à 65 mmHg dans un modèle porcin / Introduction : Refractory cardiac arrest is defined by the absence of the return of spontaneous circulation (ROCS) within 30 minutes of cardiopulmonary resuscitation (CRP) under medical supervision. ExtraCorporeal membrane oxygenation (ECMO) is an emerging alternative therapy in this population. The post extracorporeal cardiopulmonary resuscitation (ECPR) hemodynamic state is a complex entity and the critical care management in the first hours following ECMO implantation is not well defined. This study was designed to assess the effect of two veno-arterial Extracorporeal Membrane Oxygenation (ECMO) blood-flow strategies in an experimental model of ECPR (extracorporeal cardio-pulmonary resuscitation) on macrocirculatory, metabolic and microcirculatory parameters in the first six hours of ECMO initiation. Material and methods : Cardiac arrest was induced in 18 pigs by surgical ligature of the left descending coronary artery. ECPR was initiated after 40 minutes of low-flow with an ECMO blood-flow of 30-35 ml.kg-1.min-1 (low-blood-flow group, LBF) or 65-70 ml.kg-1.min-1 (standard-blood-flow group, SBF), with the same mean arterial pressure target (65 mmHg). Macrocirculatory and metabolic parameters were assessed by lactate clearance and carotid blood-flow. Microcirculatory parameters were assessed by sublingual microcirculation with Sidestream Dark Field (SDF) imaging and peripheral Near-InfraRed Spectrometry (NIRS). Inflammatory cytokine levels were measured with a multiplexed ELISA-based array platform. Results : There was no between-group difference at baseline and at ECMO initiation (H0). Lactate clearance at H6 was lower in LBF compared to SBF (6.67[-10.43-18.78] vs. 47.41[19.54, 70.69] %, p=0.04). carotid blood flow was significantly lower (p<0.005) during the last four hours despite similar mean arterial pressure levels. For microcirculatory parameters, SDF and NIRS parameters were transitorily impaired at H3 in LBF. IL-6 cytokine level was significantly higher in LBF at the end of the experiment. Conclusion: In an experimental porcine model of refractory cardiac arrest treated by ECMO, a low-blood-flow strategy during the first six hours of resuscitation was associated with lower lactate clearance and lower cerebral blood-flow with no benefits on ischemia-reperfusion parameters

A importância dada aos parâmetros de qualidade da água, principalmente nas últimas décadas justifica a realização desse trabalho, que investiga a relação de alguns parâmetros turbulentos com o coeficiente de reaeração, 'K IND.2', através de duas técnicas distintas de medida: o método ótico e a técnica fotográfica. O método ótico relaciona a intensidade de luz recebida por um diodo imerso na água com a velocidade vertical no ponto onde é realizada a medida; os dados coletados pelo método ótico constituem um sinal elétrico que contém a informação da deformação da superfície. A partir desse sinal determinam-se os seguintes parâmetros: valor RMS da tensão, função de autocorrelação, macroescala e espectro. Os resultados obtidos por esse método são funções temporais. O método fotográfico consiste em obter fotografias de partículas iluminadas sobre a superfície livre da água, com tempo de exposição conhecido. O método foi melhorado pela adição de dois disparos de flash, para permitir a determinação da variação da velocidade. As fotos são digitalizadas em um software gráfico e os valores obtidos são interpolados através do método Kriging de interpolação. O método fotográfico é uma função espacial. A partir do método fotográfico são determinados os seguintes parâmetros: valor RMS, função de autocorrelação e espectro. Os resultados obtidos permitem a comparação quantitativa entre as funções de autocorrelação e espectros, e a relação entre o coeficiente de reaeração, 'K IND.2' e o valor RMS obtidos pela sonda e pela foto / The importance given to the parameters of quality of the water, mainly in the last decades, it justifies the accomplishment of this work, that it investigates the relation of some turbulent parameters with the re-aeration coefficient, 'K IND.2', through two different techniques of measure: the optic method and the photographic technique. The optic method relates the intensity of the light received by a immersed diode in the water with the vertical speed at the point where the measure is accomplished; the data collected by the optic method are electric signs that contains the information of the deformation of the surface. Thereafter this signs, the following parameters are determined: RMS value of tension, self-correlation function, macro-scale and spectrum. The results obtained for this method they are temporal functions. The photographic method consists of obtaining pictures of illuminated particles on the free surface of the water, with time of exhibition known. The method was improved by addition of two shots of flashes, to allow the determination of the variation of the speed. The pictures, then, are digitized in a graphic software and the values obtained are interpolated through the Kriging interpolation method. The photographic method is a spatial function. With the photographic method may be determined the following parameters: RMS value, self-correlation function and spectrum. The results obtained may provide the quantitative comparison between the self-correlation function and spectrum; and also the relation between the re-aeration coefficient, 'K IND.2', and the RMS value, that are obtained through the probe and the picture

O objetivo deste trabalho foi avaliar os efeitos do monossialogangliosídio (GM1), da câmara de oxigenoterapia hiperbárica e de ambos no tratamento da lesão medular experimental em ratos. Trinta e dois ratos Wistar com lesão medular foram divididos em 4 grupos: um grupo recebeu o monossialogangliosídio (GM1), um segundo foi submetido à oxigenoterapia hiperbárica, um terceiro recebeu os dois tratamentos e um quarto não recebeu tratamento (controle). Não houve diferença significativa entre os grupos na análise histológica, em todas as variáveis (necrose, hemorragia, hiperemia e degeneração cística, p>0,06). Também não houve nenhuma diferença na comparação entre os lados direito e esquerdo nos testes funcionais (p>0,06 para todos). Não foram encontradas diferenças nos testes motores, na comparação entre os grupos após 2, 7 21 e 28 dias de lesão medular. Mas, na avaliação após 14 dias, o Grupo 3, o qual recebeu a terapia combinada, mostrou um escore BBB significantemente maior que os outros grupos (p=0,015). Na avaliação de 28 dias, houve uma tendência dos Grupos 1 (GM1) e 3 (terapia combinada) apresentarem um escore BBB maior que o do Grupo 4 (controle), embora sem significância estatística (p=0,057). Concluiu-se que, quanto aos índices motores, a utilização do GM-1 tem efeito benéfico, embora sem diferença estatisticamente significante e que o efeito benéfico do GM-1 é antecipado através da utilização concomitante da oxigênio terapia hiperbárica. / The objectives were to evaluate the effect of GM1 ganglioside, hyperbaric oxygen, and both in combination, in the treatment of experimental spinal cord lesions in rats. Thirty-two Wistar rats with spinal cord lesions were divided into four groups: one group received GM1 ganglioside, one was submitted to hyperbaric oxygen therapy, the third received both treatments, and the fourth received no treatment (control). There were no significant differences between the groups in the histological analysis, for any of the variables (necrosis, hemorrhage, hyperemia, cystic degeneration, p > 0.06). Neither were there any significant differences in the comparison of left and right sides in the functional tests (p > 0.06 for all). No significant differences were found in the locomotor ratings, in the comparison of groups at 2 days, 7 days, 21 days and 28 days after the surgical procedure. However, in the evaluation on day 14, Group 3, which received the combined therapy, showed a significantly higher BBB score than the other groups (p = 0.015). In the evaluation on day 28, there was a trend to Group 1 (GM1) and 3 (combined therapy) showed a higher BBB score than the group 4 (control), but with no significance (p=0,057). In conclusion, the is a benefit in the use of GM1 ganglioside, but with no significance and the therapeutic effect of GM1 in locomotor evaluation of rats submitted to spinal cord lesion is anticipated by hyperbaric oxygen therapy.

A anemia é a causa primária de indicação de transfusão de glóbulos vermelhos e é especialmente prevalente e esperada em centros de terapia intensiva. Os benefícios das transfusões incluem aumento da oxigenação tissular, diminuição de hemorragias, tratamento da anemia e da hipovolemia. Os poucos estudos publicados têm demonstrado variações significativas nas práticas de transfusões de glóbulos vermelhos e o melhor momento para a indicação destas transfusões ainda não foi identificado. Objetivos Descrever a população de crianças que recebeu transfusão de glóbulos vermelhos com relação à faixa etária, concentração de hemoglobina (Hb) e taxa de hematócrito (HT), uso de saturação venosa central de oxigênio (SvO2), lactato sérico arterial, procedimentos terapêuticos e presença de disfunção de múltiplos órgãos e sistemas (DMOS). Métodos Estudo retrospectivo observacional, realizado no Centro de Terapia Intensiva Pediátrico (CTIP) do Instituto da Criança da Faculdade de Medicina da Universidade de São Paulo, em 2004, com crianças que receberam transfusão de glóbulos vermelhos. Descrevemos o número de pacientes, faixa etária, motivo de internação no CTIP, presença de doença de base e de DMOS, uso de SvO2, lactato sérico arterial, Hb, HT e procedimentos terapêuticos (ventilação mecânica, drogas vasoativas e métodos de substituição renal). Resultados A transfusão de glóbulos vermelhos foi realizada em 50% dos pacientes internados. A idade mediana foi de 18 meses e o principal motivo de internação foi insuficiência respiratória (35% dos casos). Doença de base estava presente em 84% dos casos e DMOS em 47% dos casos. Foram realizadas coletas de Hb e HT em todos os casos e de SvO2 e lactato sérico arterial em 24% dos casos. A mediana de Hb pré-transfusional foi de 7,8 g/dl. Os pacientes transfundidos estavam sendo submetidos a algum procedimento terapêutico em 82% dos casos. Conclusões São realizadas transfusões de glóbulos vermelhos em todas as idades. A concentração de hemoglobina e a taxa de hematócrito são os principais dados utilizados para a indicação destas transfusões. O lactato sérico arterial e a SvO2 são pouco utilizados para se indicar a transfusão de glóbulos vermelhos. A maioria dos pacientes transfundidos recebe algum procedimento terapêutico e, em muitos casos, são realizadas transfusões em pacientes que apresentam DMOS. / Anemia is the primary cause of red blood cell transfusions and it is especially prevalent and even expected in critical care settings. The benefits of red blood cell transfusions include increase in tissue oxygenation, treatment of anemia and hypovolemia. There are substantial variations in transfusion practices and the optimal transfusion practice for various types of critically ill patients with anemia has not been established. Objectives To describe the clinical, hematological and therapeutic characteristics of children who received red blood cell transfusion in a Pediatric Intensive Care Unit (PICU). Methods Retrospective observational study, performed in the PICU of ?Instituto da Criança?, Medicine School of University of São Paulo, in 2004, with one hundred children who received red blood cell transfusion. We described the number of patients included in the study, age, reason for admission to the PICU, presence of baseline illness and multiple organ system failure (MODS), use of oxygen central venous saturation, arterial lactate, hemoglobin concentration (Hb), hematocrit (HT) and therapeutics procedures (mechanical ventilation, use of vasoactive drugs and renal replacement methods). Results The red blood cell transfusion occurred in 50% of the patients. Median age of patients was 18 months. The most common reason for admission was respiratory failure, in 35% of the cases. Baseline illness was present in 84% of the cases and MODS in 47% of the cases. Hb and HT were collected in all the cases and oxygen central venous saturation and arterial lactate were collected in 24% of the cases. The median pre-transfusion hemoglobin concentration was 7.8 g/dl. The patients were receiveing therapeutics procedures in 82% of the cases. Conclusions Red blood cell transfusions are performed in children of all ages. The hemoglobin concentration and hematocrit are the main reason to indicate the red blood cell transfusion. Arterial lactate and oxygen central venous saturation are rarely utilized. The majority of patients receive therapeutic procedures and, in many cases, patients with MODS are given red blood cell transfusions.

Procurou-se confirmar a existência de alterações estruturais em corpos carotídeos de ratos expostos à hiperóxia que pudessem explicar a atenuação da resposta fisiológica à hipóxia nessas condições descrita na literatura. Também testamos a hipótese de haver um desvio de fluxo sanguíneo para os capilares intraglômicos em situações de hiperoxigenação hiperbárica.15 ratos machos Wistar adultos foram divididos em 3 grupos e expostos a 2,4 ATA por 6 horas, a 3,0 ATA por 6 horas e a ar ambiente (grupo controle). Os resultados obtidos através de análise histológica e morfométrica mostraram: a) nenhuma alteração da arquitetura dos corpos carotídeos, mas as células expostas à dose mais elevada apresentaram-se com citoplasma desarranjado, confirmado pela microscopia eletrônica; b) um aumento significativo da densidade volumétrica de capilares preenchidos por hemácias, mas não do estroma intersticial, no grupo exposto à dose mais elevada de O2 c) uma vasoconstricção significativa das arteríolas maiores em todas as doses de oxigênio empregadas no estudo e das arteríolas menores na dose mais elevada de O2; d) variações significativas na proporção das variantes de células glômicas no grupo exposto a menor dose de O2; e) mitocôndrias com poucas cristas, tanto nas células glômicas quanto nas terminações nervosas, embora nas primeiras apresentem-se bem deformadas; f) proliferação membranosa citoplasmática com aumento de REG e Golgi nas células glômicas e sustentaculares. Esses resultados sugerem um desvio do fluxo dos vasos mais calibrosos em direção aos capilares intraglômicos, confirmando nossa hipótese inicial e indicam que o oxigênio, dependendo da dose utilizada, exerce um efeito tóxico importante sobre os corpos carotídeos, com alterações significativas da ultraestrutura das células glômicas e terminações nervosas. / We sough to confirm the existence of structural alterations in rat carotid bodies exposed to hyperoxia that could explain the attenuation of the ventilatory hypoxic drive (HD) by hyperoxic conditions described in the literature. We also tested the hypothesis of there being a deviation of blood flow toward intraglomic capillaries in situations of hyperbaric oxygenation (HBO).15 adult male Wistar rats were divided in 3 groups and exposed to O2 at 2.4 ATA for 6 hours, at 3.0 ATA for 6 hours and to air at 1.0 ATA (control group). The results obtained through histological and morphometric analysis showed: a) no alteration in the architecture of the carotid bodies, but the cytoplasm of the cells exposed to the highest dose were disarranged, a feature confirmed by electron microscopy; b) a significant increase in volume density of capillaries filled out by red blood cells but not of interstitial stroma in the group exposed to O2 at the highest dose; c) a significant vasoconstriction of larger arterioles in all doses of oxygen employed in the study and of smaller arterioles at the highest dose of O2; d) significant variations in the proportion of glomic cell variants in the group exposed to the lowest dose of O2; e) mitochondria with few cristae, so in glomic cells as in nerve-endings, although in the former they were very deformed; f) cytoplasmic membranous proliferation with an increase of endoplasmic reticulum and Golgi apparatus in glomic and sustentacular cells. These results suggest a deviation of blood flow from more calibrated vessel toward intraglomic capillaries, confirming our initial hypothesis and indicate that oxygen, depending on the dose used, exerts an important toxic effect on rat carotid body with significant alterations of glomic cell and nerve-endings ultrastructure.

A anemia é a causa primária de indicação de transfusão de glóbulos vermelhos e é especialmente prevalente e esperada em centros de terapia intensiva. Os benefícios das transfusões incluem aumento da oxigenação tissular, diminuição de hemorragias, tratamento da anemia e da hipovolemia. Os poucos estudos publicados têm demonstrado variações significativas nas práticas de transfusões de glóbulos vermelhos e o melhor momento para a indicação destas transfusões ainda não foi identificado. Objetivos Descrever a população de crianças que recebeu transfusão de glóbulos vermelhos com relação à faixa etária, concentração de hemoglobina (Hb) e taxa de hematócrito (HT), uso de saturação venosa central de oxigênio (SvO2), lactato sérico arterial, procedimentos terapêuticos e presença de disfunção de múltiplos órgãos e sistemas (DMOS). Métodos Estudo retrospectivo observacional, realizado no Centro de Terapia Intensiva Pediátrico (CTIP) do Instituto da Criança da Faculdade de Medicina da Universidade de São Paulo, em 2004, com crianças que receberam transfusão de glóbulos vermelhos. Descrevemos o número de pacientes, faixa etária, motivo de internação no CTIP, presença de doença de base e de DMOS, uso de SvO2, lactato sérico arterial, Hb, HT e procedimentos terapêuticos (ventilação mecânica, drogas vasoativas e métodos de substituição renal). Resultados A transfusão de glóbulos vermelhos foi realizada em 50% dos pacientes internados. A idade mediana foi de 18 meses e o principal motivo de internação foi insuficiência respiratória (35% dos casos). Doença de base estava presente em 84% dos casos e DMOS em 47% dos casos. Foram realizadas coletas de Hb e HT em todos os casos e de SvO2 e lactato sérico arterial em 24% dos casos. A mediana de Hb pré-transfusional foi de 7,8 g/dl. Os pacientes transfundidos estavam sendo submetidos a algum procedimento terapêutico em 82% dos casos. Conclusões São realizadas transfusões de glóbulos vermelhos em todas as idades. A concentração de hemoglobina e a taxa de hematócrito são os principais dados utilizados para a indicação destas transfusões. O lactato sérico arterial e a SvO2 são pouco utilizados para se indicar a transfusão de glóbulos vermelhos. A maioria dos pacientes transfundidos recebe algum procedimento terapêutico e, em muitos casos, são realizadas transfusões em pacientes que apresentam DMOS. / Anemia is the primary cause of red blood cell transfusions and it is especially prevalent and even expected in critical care settings. The benefits of red blood cell transfusions include increase in tissue oxygenation, treatment of anemia and hypovolemia. There are substantial variations in transfusion practices and the optimal transfusion practice for various types of critically ill patients with anemia has not been established. Objectives To describe the clinical, hematological and therapeutic characteristics of children who received red blood cell transfusion in a Pediatric Intensive Care Unit (PICU). Methods Retrospective observational study, performed in the PICU of ?Instituto da Criança?, Medicine School of University of São Paulo, in 2004, with one hundred children who received red blood cell transfusion. We described the number of patients included in the study, age, reason for admission to the PICU, presence of baseline illness and multiple organ system failure (MODS), use of oxygen central venous saturation, arterial lactate, hemoglobin concentration (Hb), hematocrit (HT) and therapeutics procedures (mechanical ventilation, use of vasoactive drugs and renal replacement methods). Results The red blood cell transfusion occurred in 50% of the patients. Median age of patients was 18 months. The most common reason for admission was respiratory failure, in 35% of the cases. Baseline illness was present in 84% of the cases and MODS in 47% of the cases. Hb and HT were collected in all the cases and oxygen central venous saturation and arterial lactate were collected in 24% of the cases. The median pre-transfusion hemoglobin concentration was 7.8 g/dl. The patients were receiveing therapeutics procedures in 82% of the cases. Conclusions Red blood cell transfusions are performed in children of all ages. The hemoglobin concentration and hematocrit are the main reason to indicate the red blood cell transfusion. Arterial lactate and oxygen central venous saturation are rarely utilized. The majority of patients receive therapeutic procedures and, in many cases, patients with MODS are given red blood cell transfusions.

<p>Type 1 diabetes mellitus affects millions of people worldwide. Islet transplantation is a minimal invasive surgical procedure that restores euglycemia and halts the progression of diabetic complications. However, despite transplantation of islets from multiple donors most patients reverse to hyperglycemia within five years. New strategies to improve long-term outcome of islet transplantation are indispensable. This thesis studied differences in the microvasculature between endogenous and transplanted pancreatic islets, and investigated means to improve islet graft revascularization and function. Islet graft microvessels were similar to endogenous islets responsive to adenosine, angiotensin II and nitric oxide (NO). Recipient hyperglycemia induced a higher basal islet graft blood flow, which also was less dependent on NO than in normoglycemic recipients. Transplantation of freshly isolated instead of cultured islets improved graft revascularization, oxygenation and function. Pretreatment of islets with vascular endothelial growth factor decreased their expression of matrix metalloproteinase-9 (MMP-9) and impaired graft revascularization. Moreover, MMP-9 pretreatment <i>per se</i> improved graft revascularization. <i>In vivo</i>, 20-25% of all endogenous rat islets was low oxygenated (pO₂ <10 mmHg). Changes in the islet mass, by means of whole-pancreas transplantation, doubled the fraction of low oxygenated islets in the endogenous pancreas of transplanted animals, whereas this fraction almost completely disappeared after a 60% partial pancreatectomy. Interestingly, oxygenation was related to metabolism, since well oxygenated islets <i>in vivo</i> had 50% higher leucine-dependent protein biosynthesis, which includes (pro)insulin biosynthesis. In intraportally transplanted islets, the low oxygenated fraction of islets was markedly increased one day post-transplantation, and the oxygenation remained low following revascularization. In summary, these data suggest that a better revascularization of transplanted islets can improve graft function. Furthermore, the oxygenation and metabolism of endogenous islets is tightly regulated. This regulation seems to be disturbed following transplantation, which may contribute to long-term islet graft failure. </p>

In the past decades, tumors have progressively been perceived as highly integrated systems in which the genetically unstable tumor cells and the genetically stable host cells cooperate to promote tumor growth. This view suggests that, beside tumor cells (that are targeted by conventional anticancer treatments such as radio- and chemotherapy), host cells within the tumor microenvironment can be targeted by antitumor therapy. Such alternative strategies are strongly supported by the need to overcome several limitations of the conventional therapies targeting tumor cells, such as collateral toxicity due to lack of tumor selectivity, limited tumor accessibility, and the selection of treatment-resistant variants. By contrast to tumor cells, the genetically stable host cells should not develop resistance to treatments. In this context, the observation that tumor growth is fundamentally dependent on the onset of a private tumor neovasculature (tumor angiogenesis) has revolutionized the field of cancer research. Several treatments have been developed aimed to prevent tumor angiogenesis (anti-angiogenic strategies) or to erase the existent tumor vasculature (anti-vascular approaches) supporting the survival and growth of thousands of tumor cells. However, although such therapies achieved cancer cure in animal models, they turned out to be rather inefficient when tested in patients. This can be attributed to differences in the angiogenic status between fast-growing animal tumors and slow-growing human tumors at the time of clinical detection. Another reading of the above-mentioned observations is that anticancer treatments could benefit from interventions aimed at increasing their efficiency. For instance, radiotherapy could benefit from tumor reoxygenation while a decrease in tumor interstitial pressure could facilitate tumor accessibility to circulating agents. In this context, the mature vasculature is an attractive target since it controls tumor blood supply and is highly accessible for therapy. Therefore, strategies aimed at exploiting its functional reactivity by inducing vasorelaxation have the potential to improve tumor perfusion/drug delivery and oxygenation/radiosensitivity. To be exploited in the clinics, such pro-vascular approaches have to fulfill essential requirements. First, they need to achieve high selectivity for tumor vessels. It should prevent systemic toxicity as well as the stealing of the blood flow towards the peripheral vasculature. Second, vasodilation has to be transient, so that the tumor should not take advantage of an increased energetic supply to grow faster. Third, the therapeutic effects have to be achieved in several tumor types and in different host strains to gain a wide therapeutic range of applicability. Finally, vasomodulation has to be achieved with interventions relevant to the clinical situation, ensuring direct therapeutic significance. However, the therapeutic exploitation of agents modulating tumor perfusion was generally hampered by confounding effects on the systemic blood pressure. In our studies, we have documented that this lack of tumor selectivity can be overcome by identifying vasomodulatory pathways that are selectively altered within the tumor microenvironment, allowing selective vasomodulatory interventions. According to the criteria detailed above, to identify a differential tumor vascular reactivity, we had to work with mice models of mature tumor vascularization. We reasoned that preexisting host arterioles in mice, if coopted, should retain architectural characteristics (such as a muscular coat) necessary for functional reactivity but also be influenced by the tumor microenvironment at both molecular and functional levels. To gain in reproducibility, this model was developed by injecting syngeneic tumor cells in the vicinity of the saphenous arteriole (i.e., a collateral branch of the femoral artery) in the rear leg of mice. With tumor growth, this arteriole was progressively included in the tumor cortex (coopted), with side branches running deeply into tumors. This model was developed using several tumors and mice strains. It provides the unique advantage to allow the easy identification and isolation of mature tumor vessels from fast-growing animal tumors. To evaluate differential vasoreactivity in those tumor-coopted vessels, we adapted pressure myography, a device initially dedicated to the study of the reactivity of coronary arterioles (see annex 1). In our hands, the unprecedented application of pressure myography to the study of small tumor vessels proved to be very efficient. Indeed, this technique not only served us to confirm that arterioles remain sensitive to vasomodulation under tumor cooption, but also allowed us to evidence two major adaptations of host vessels to the tumor microenvironment: the acquisition of an ET-1-mediated basal constrictive tone and a defect in the vasodilatory NO pathway. Furthermore, we used pressure myography to identify and characterize vasomodulatory strategies exploiting these differential reactivities. More particularly, we showed that both BQ123 (an ETA inhibitor) and ionizing radiations (that restored a functional NO pathway) promoted the vasodilation of the tumor-coopted vessels. In vivo, we verified that these strategies fulfilled the essential requirements of pro-vascular approaches: tumor selectivity, transient effects, broad range of applicability, and therapeutic significance in clinically relevant regimens. This latter study led us to further explore the effects of radiotherapy on the status of the tumor vasculature. Hence, we showed that fractionated radiotherapy induced tumor angiogenesis, thereby providing a rationale to combine radiotherapy to anti-angiogenic therapies.

Type 1 diabetes mellitus affects millions of people worldwide. Islet transplantation is a minimal invasive surgical procedure that restores euglycemia and halts the progression of diabetic complications. However, despite transplantation of islets from multiple donors most patients reverse to hyperglycemia within five years. New strategies to improve long-term outcome of islet transplantation are indispensable. This thesis studied differences in the microvasculature between endogenous and transplanted pancreatic islets, and investigated means to improve islet graft revascularization and function. Islet graft microvessels were similar to endogenous islets responsive to adenosine, angiotensin II and nitric oxide (NO). Recipient hyperglycemia induced a higher basal islet graft blood flow, which also was less dependent on NO than in normoglycemic recipients. Transplantation of freshly isolated instead of cultured islets improved graft revascularization, oxygenation and function. Pretreatment of islets with vascular endothelial growth factor decreased their expression of matrix metalloproteinase-9 (MMP-9) and impaired graft revascularization. Moreover, MMP-9 pretreatment per se improved graft revascularization. In vivo, 20-25% of all endogenous rat islets was low oxygenated (pO2 &lt;10 mmHg). Changes in the islet mass, by means of whole-pancreas transplantation, doubled the fraction of low oxygenated islets in the endogenous pancreas of transplanted animals, whereas this fraction almost completely disappeared after a 60% partial pancreatectomy. Interestingly, oxygenation was related to metabolism, since well oxygenated islets in vivo had 50% higher leucine-dependent protein biosynthesis, which includes (pro)insulin biosynthesis. In intraportally transplanted islets, the low oxygenated fraction of islets was markedly increased one day post-transplantation, and the oxygenation remained low following revascularization. In summary, these data suggest that a better revascularization of transplanted islets can improve graft function. Furthermore, the oxygenation and metabolism of endogenous islets is tightly regulated. This regulation seems to be disturbed following transplantation, which may contribute to long-term islet graft failure.