Outflow occlusion for circulatory arrest in dogs

The purpose of this study was to evaluate the possibility of producing circulatory arrest by occlusion of the pulmonary trunk as an alternative to the venous inflow occlusion through the left hemithorax. Eight healthy mongrel dogs were divided in two groups. Group I underwent 4 minutes of outflow occlusion and Group II was submitted to 8 minutes of circulatory arrest. Outflow occlusion was performed through left thoracotomy and pericardiotomy by passing a Rumel tourniquet around the pulmonary trunk. Physical examination, electrocardiography, echocardiography, blood gas analyses, hemodynamic, and oxygen transport variables were obtained before and after the procedure. The dogs from Group I did not have any clinical, electrocardiographic, echocardiographic, or hemo-dynamic abnormalities after anesthetic recover. In the Group II, only one dog survived, which had no clinical, electrocardiographic, or echocardiographic abnormalities. In this last dog, just after releasing the occlusion, it was detected increases in the following parameters: heart rate (HR), systolic, diastolic and mean arterial blood pressure (SAP; DAP; MAP), pulmonary artery pressure (PAP), pulmonary wedge pressure (PWP), central venous pressure (CVP), cardiac output (CO), systolic index (SI), cardiac index (CI), left and right ventricular stroke work (LVSW; RVSW), oxygen delivery index (DO2), oxygen consumption index (VO2), and oxygen extraction (O2 ext). Moreover, the oxygen content of arterial and mixed venous blood (CaO2; CvO2), and the arterial and mixed venous partial pressure of oxygen (PaO2; PvO2) were decreased 5 minutes after circulatory arrest. Outflow occlusion is a feasible surgical procedure for period of 4 minutes of circulatory arrest.

AT1-receptor mediated vascular damage in myocardium, kidneys and liver in rats

The systemic aspect of vascular damage induced by angiotensin II (ANG II) has been poorly explored in the literature. Considering the presence of ANG II and its specific receptor AT1, in several organs, all tissues might be potentially affected by its effects. The aims of this study were: To evaluate the early histological changes in the heart, liver and kidneys, produced by ANG II infusion, to evaluate the protective effect of losartan. Wistar rats were distributed into three groups: control (no treatment), treated with ANG II, and treated with ANG II + losartan. ANG II was continuously infused over 72 hours by subcutaneous osmotic pumps. Histological sections of the myocardium, kidneys and liver were stained and observed for the presence of necrosis. There were ANG II-induced perivascular inflammation and necrosis of the arteriolar wall in the myocardium, kidney, and liver by, which were partially prevented by losartan. There was no significant correlation between heart and kidney damage. Tissue lesion severity was lower than that of vascular lesions, without statistical difference between groups. ANG II causes vascular injury in the heart, kidneys and liver, indicating a systemic vasculotoxic effect; the mechanisms of damage/protection vary depending on the target organ; perivascular lesions may occur even when anti-hypertensive doses of losartan are used.

Viabilidade celular da fração mononuclear da medula óssea e fração vascular estromal do tecido adiposo de equinos após o processo de congelamento e descongelamento

Cinco cavalos adultos foram submetidos à coleta de medula óssea do esterno e de tecido adiposo da região glútea. As amostras foram processadas para obtenção da fração mononuclear da medula óssea e fração vascular estromal do tecido adiposo, o número de células obtidas e a viabilidade celular foram determinados. Em seguida, realizou-se o congelamento das amostras em solução contendo 20% de soro fetal bovino e 10% de dimetilsulfóxido. Depois de um mês, realizou-se o descongelamento das amostras e a viabilidade celular foi novamente mensurada. Os resultados revelaram que as técnicas utilizadas tanto para coleta de medula óssea quanto de tecido adiposo em equinos são simples, rápidas e seguras. As metodologias adotadas para o processamento das amostras foram eficientes, obtendo-se aproximadamente 95% de viabilidade celular. Após o descongelamento, a viabilidade média das amostras de células mononucleares da medula óssea foi de 86% e da fração vascular estromal do tecido adiposo de 64%. Frente à importância da terapia celular na clínica médica de equinos, concluiu-se que é necessária a realização de mais estudos, visando padronizar uma técnica de criopreservação que mantenha a integridade das células da fração mononuclear da medula óssea e da fração vascular estromal do tecido adiposo de equinos.

Culture of equine bone marrow mononuclear fraction and adipose tissue-derived stromal vascular fraction cells in different media

The objective of this study was to evaluate the culture of equine bone marrow mononuclear fraction and adipose tissue - derived stromal vascular fraction cells in two different cell culture media. Five adult horses were submitted to bone marrow aspiration from the sternum, and then from the adipose tissue of the gluteal region near the base of the tail. Mononuclear fraction and stromal vascular fraction were isolated from the samples and cultivated in DMEM medium supplemented with 10% fetal bovine serum or in AIM-V medium. The cultures were observed once a week with an inverted microscope, to perform a qualitative analysis of the morphology of the cells as well as the general appearance of the cell culture. Colony-forming units (CFU) were counted on days 5, 15 and 25 of cell culture. During the first week of culture, differences were observed between the samples from the same source maintained in different culture media. The number of colonies was significantly higher in samples of bone marrow in relation to samples of adipose tissue.

Sistema vascular e controle do desenvolvimento de perfilhos em cereais de estação fria

As causas da ausência e/ou baixa sobrevivência dos perfilhos foram estudadas pela conexão vascular entre os perfilhos e o resto da planta. Foram conduzidos ensaios em telado e câmara de crescimento com genótipos de trigo, IPF-49865 (unicolmo), EMBRAPA-16 e BR-23 (multicolmo), de aveia, UFRGS-10 e UFRGS-15 e de cevada, FM-519. Avaliou-se o desenvolvimento foliar do colmo principal e dos perfilhos primários, o número de perfilhos primários e secundários, o número de perfilhos férteis e a vascularização e conexão vascular dos perfilhos com o resto da planta. A conexão vascular poderia explicar a inibição dos perfilhos pois, todos estavam conectados vascularmente com o resto da planta.

Composição florística e estrutura do componente epifítico vascular em floresta da planície litorânea na Ilha do Mel, Paraná, Brasil

O estudo das epífitas vasculares foi realizado em uma área de 3.000 m² de floresta na planície litorânea na Ilha do Mel (25°30’ S 48°23’ W); o levantamento qualitativo foi realizado em toda a área; para o estudo quantitativo, foram sorteados 100 forófitos, divididos em intervalos de 2 m a partir do solo. Em cada intervalo, registraram-se todas as espécies epifíticas ocorrentes, sendo estimado o valor de importância epifítico a partir das freqüências nos estratos, nos forófitos e nas espécies forofíticas. No levantamento total, foram encontradas 77 espécies (16 de Pteridophyta, 53 de Liliopsida e 8 de Magnoliopsida), das quais 70 foram amostradas nos forófitos analisados. As famílias mais ricas foram Orchidaceae, Bromeliaceae e Polypodiaceae e os gêneros foram Vriesea, Encyclia e Maxillaria. A área com maior similaridade florística com este estudo localiza-se no Município de Torres (RS). As espécies mais importantes quantitativamente foram Microgramma vaccinifolia, Codonanthe gracilis, Epidendrum latilabre e E. rigidum. As espécies amostradas foram agrupadas em três categorias quanto à preferência por intervalos de altura: exclusivas, preferenciais e indiferentes. O número de ocorrências de epífitos em um mesmo forófito variou de 1 a 35, enquanto o número de espécies variou de 1 a 21 (médias 14 ± 7,6 e 10 ± 4,6, respectivamente). Os primeiros estratos (0-2 m, 2-4 m e 4-6 m) foram os mais ricos em espécies epifíticas. As espécies forofíticas com maior número de ocorrências foram Andira fraxinifolia e Ternstroemia brasiliensis, e com maior número de espécies Ocotea pulchella e Guapira opposita. Quanto à fidelidade sobre espécies forofíticas, foram encontradas espécies epifíticas exclusivas, preferenciais e generalistas, esta incluindo a maioria das espécies amostradas.

Florística e estrutura do componente epifítico vascular em floresta ombrófila mista aluvial do rio Barigüi, Paraná, Brasil

O estudo das epífitas vasculares foi realizado em floresta ombrófila mista aluvial do rio Barigüi, município de Araucária (25º34' S e 49º20' W); o levantamento qualitativo foi realizado em uma área aproximada de 8,6 ha. Para o estudo quantitativo foram sorteados 110 forófitos, divididos em três estratos (fuste baixo, fuste alto e copa). Em cada estrato foram atribuídas notas de abundância para as espécies epifíticas ocorrentes. No levantamento florístico foram encontradas 49 espécies (16 de Pteridophyta, 23 de Liliopsida e 10 de Magnoliopsida), das quais 34 foram observadas na amostragem quantitativa. As famílias e gêneros mais ricos foram: Orchidaceae, Polypodiaceae e Bromeliaceae, e Pleurothallis, Tillandsia e Oncidium. As áreas de maior similaridade florística com este estudo localizam-se no município de Curitiba (PR). As espécies mais importantes quantitativamente foram Microgramma squamulosa (Kaulf.) de la Sota, Pleopeltis angusta Humb. & Bonpl. ex Kunth, Peperomia catharinae Miq. e Polypodium hirsutissimum Raddi. Em um mesmo forófito, o número de espécies ocorrentes variou de zero a 13, enquanto a soma das notas de abundância para as epífitas variou de zero a 24. A copa apresentou a maior abundância de epífitos, seguido do fuste alto e do fuste baixo.

Community structure of vascular plants in treefall gaps and fire-disturbed habitats in the Atlantic rainforest, southern Bahia, Brazil

The effects of disturbances on plant community structure in tropical forests have been widely investigated. However, a majority of these studies examined only woody species, principally trees, whereas the effects of disturbances on the whole assemblage of vascular plants remain largely unexplored. At the present study, all vascular plants < 5m tall were surveyed in four habitats: natural treefall gaps, burned forest, and their adjacent understorey. The burned area differed from the other habitats in terms of species composition. However, species richness and plant density did not differ between burned area and the adjacent understorey, which is in accordance to the succession model that predict a rapid recovery of species richness, but with a different species composition in areas under moderate disturbance. The treefall gaps and the two areas of understorey did not differ among themselves in terms of the number of individuals, number of species, nor in species composition. The absence of differences between the vegetation in treefall gaps and in understorey areas seems to be in agreement with the current idea that the species present in treefall gaps are directly related to the vegetation composition before gap formation. Only minimal differences were observed between the analyses that considered only tree species and those that considered all growth habits. This suggests that the same processes acting on tree species (the best studied group of plants in tropical forests) are also acting on the whole assemblage of vascular plants in these communities.

Variation in nitrogen use strategies and photosynthetic pathways among vascular epiphytes in the Brazilian Central Amazon

The variation in nitrogen use strategies and photosynthetic pathways among vascular epiphyte families was addressed in a white-sand vegetation in the Brazilian Central Amazon. Foliar nitrogen and carbon concentrations and their isotopic composition (δ15N and δ13C, respectively) were measured in epiphytes (Araceae, Bromeliaceae and Orchidaceae) and their host trees. The host tree Aldina heterophylla had higher foliar N concentration and lower C:N ratio (2.1 ± 0.06% and 23.6 ± 0.8) than its dwellers. Tree foliar δ15N differed only from that of the orchids. Comparing the epiphyte families, the aroids had the highest foliar N concentration and lowest C:N ratios (1.4 ± 0.1% and 34.9 ± 4.2, respectively). The orchids had more negative foliar δ15N values (-3.5 ± 0.2‰) than the aroids (-1.9 ± 0.7‰) and the bromeliads (-1.1 ± 0.6‰). Within each family, aroid and orchid taxa differed in relation to foliar N concentrations and C:N ratios, whereas no internal variation was detected within bromeliads. The differences in foliar δ15N observed herein seem to be related to the differential reliance on the available N sources for epiphytes, as well as to the microhabitat quality within the canopy. In relation to epiphyte foliar δ13C, the majority of epiphytes use the water-conserving CAM-pathway (δ13C values around -17‰), commonly associated with plants that live under limited and intermittent water supply. Only the aroids and one orchid taxon indicated the use of C3-pathway (δ13C values around -30‰).

Calcium handling by vascular myocytes in hypertension

Calcium ions (Ca2+) trigger the contraction of vascular myocytes and the level of free intracellular Ca2+ within the myocyte is precisely regulated by sequestration and extrusion mechanisms. Extensive evidence indicates that a defect in the regulation of intracellular Ca2+ plays a role in the augmented vascular reactivity characteristic of clinical and experimental hypertension. For example, arteries from spontaneously hypertensive rats (SHR) have an increased contractile sensitivity to extracellular Ca2+ and intracellular Ca2+ levels are elevated in aortic smooth muscle cells of SHR. We hypothesize that these changes are due to an increase in membrane Ca2+ channel density and possibly function in vascular myocytes from hypertensive animals. Several observations using various experimental approaches support this hypothesis: 1) the contractile activity in response to depolarizing stimuli is increased in arteries from hypertensive animals demonstrating increased voltage-dependent Ca2+ channel activity in hypertension; 2) Ca2+ channel agonists such as Bay K 8644 produce contractions in isolated arterial segments from hypertensive rats and minimal contraction in those from normotensive rats; 3) intracellular Ca2+ concentration is abnormally increased in vascular myocytes from hypertensive animals following treatment with Ca2+ channel agonists and depolarizing interventions, and 4) using the voltage-clamp technique, the inward Ca2+ current in arterial myocytes from hypertensive rats is nearly twice as large as that from myocytes of normotensive rats. We suggest that an alteration in Ca2+ channel function and/or an increase in Ca2+ channel density, resulting from increased channel synthesis or reduced turnover, underlies the increased vascular reactivity characteristic of hypertension

Effects of ouabain on vascular reactivity

Ouabain is an endogenous substance occurring in the plasma in the nanomolar range, that has been proposed to increase vascular resistance and induce hypertension. This substance acts on the a-subunit of Na+,K+-ATPase inhibiting the Na+-pump activity. In the vascular smooth muscle this effect leads to intracellular Na+ accumulation that reduces the activity of the Na+/Ca2+ exchanger and to an increased vascular tone. It was also suggested that circulating ouabain, even in the nanomolar range, sensitizes the vascular smooth muscle to vasopressor substances. We tested the latter hypothesis by studying the effects of ouabain in the micromolar and nanomolar range on phenylephrine (PE)-evoked pressor responses. The experiments were performed in normotensive and hypertensive rats in vivo, under anesthesia, and in perfused rat tail vascular beds. The results showed that ouabain pretreatment increased the vasopressor responses to PE in vitro and in vivo. This sensitization after ouabain treatment was also observed in hypertensive animals which presented an enhanced vasopressor response to PE in comparison to normotensive animals. It is suggested that ouabain at nanomolar concentrations can sensitize vascular smooth muscle to vasopressor stimuli possibly contributing to increased tone in hypertension

Reactivity of the isolated perfused rat tail vascular bed

Isolated segments of the perfused rat tail artery display a high basal tone when compared to other isolated arteries such as the mesenteric and are suitable for the assay of vasopressor agents. However, the perfusion of this artery in the entire tail has not yet been used for functional studies. The main purpose of the present study was to identify some aspects of the vascular reactivity of the rat tail vascular bed and validate this method to measure vascular reactivity. The tail severed from the body was perfused with Krebs solution containing different Ca2+ concentrations at different flow rates. Rats were anesthetized with sodium pentobarbital (65 mg/kg) and heparinized (500 U). The tail artery was dissected near the tail insertion, cannulated and perfused with Krebs solution plus 30 µM EDTA at 36oC and 2.5 ml/min and the procedures were started after equilibration of the perfusion pressure. In the first group a dose-response curve to phenylephrine (PE) (0.5, 1, 2 and 5 µg, bolus injection) was obtained at different flow rates (1.5, 2.5 and 3.5 ml/min). The mean perfusion pressure increased with flow as well as PE vasopressor responses. In a second group the flow was changed (1.5, 2, 2.5, 3 and 3.5 ml/min) at different Ca2+ concentrations (0.62, 1.25, 2.5 and 3.75 mM) in the Krebs solution. Increasing Ca2+ concentrations did not alter the flow-pressure relationship. In the third group a similar protocol was performed but the rat tail vascular bed was perfused with Krebs solution containing PE (0.1 µg/ml). There was an enhancement of the effect of PE with increasing external Ca2+ and flow. PE vasopressor responses increased after endothelial damage with air and CHAPS, suggesting an endothelial modulation of the tone of the rat tail vascular bed. These experiments validate the perfusion of the rat tail vascular bed as a method to investigate vascular reactivity

Nonspecific blockade of vascular free radical signals by methylated arginine analogues

Methylated arginine analogues are often used as probes of the effect of nitric oxide; however, their specificity is unclear and seems to be frequently overestimated. This study analyzed the effects of NG-methyl-L-arginine (L-NMMA) on the endothelium-dependent release of vascular superoxide radicals triggered by increased flow. Plasma ascorbyl radical signals measured by direct electron paramagnetic resonance spectroscopy in 25 rabbits increased by 3.8 ± 0.7 nmol/l vs baseline (28.7 ± 1.4 nmol/l, P<0.001) in response to papaverine-induced flow increases of 121 ± 12%. In contrast, after similar papaverine-induced flow increases simultaneously with L-NMMA infusions, ascorbyl levels were not significantly changed compared to baseline. Similar results were obtained in isolated rabbit aortas perfused ex vivo with the spin trap a-phenyl-N-tert-butylnitrone (N = 22). However, in both preparations, this complete blockade was not reversed by co-infusion of excess L-arginine and was also obtained by N-methyl-D-arginine, thus indicating that it is not related to nitric oxide synthase. L-arginine alone was ineffective, as previously demonstrated for NG-methyl-L-arginine ester (L-NAME). In vitro, neither L-arginine nor its analogues scavenged superoxide radicals. This nonspecific activity of methylated arginine analogues underscores the need for careful controls in order to assess nitric oxide effects, particularly those related to interactions with active oxygen species.

Integrins in vascular development

Many growth factors and their protein kinase receptors play a role in regulating vascular development. In addition, cell adhesion molecules, such as integrins and their ligands in the extracellular matrix, play important roles in the adhesion, migration, proliferation, survival and differentiation of the cells that form the vasculature. Some integrins are known to be regulated by angiogenic growth factors and studies with inhibitors of integrin functions and using strains of mice lacking specific integrins clearly implicate some of these molecules in vasculogenesis and angiogenesis. However, the data are incomplete and sometimes discordant and it is unclear how angiogenic growth factors and integrin-mediated adhesive events cooperate in the diverse cell biological processes involved in forming the vasculature. Consideration of the results suggests working hypotheses and raises questions for future research directions.

Neuronal and endothelial nitric oxide synthase gene knockout mice

Targeted disruption of the neuronal nitric oxide synthase (nNOS) and endothelial nitric oxide synthase (eNOS) genes has led to knockout mice that lack these isoforms. These animal models have been useful to study the roles of nitric oxide (NO) in physiologic processes. nNOS knockout mice have enlarged stomachs and defects in the inhibitory junction potential involved in gastrointestinal motility. eNOS knockout mice are hypertensive and lack endothelium-derived relaxing factor activity. When these animals are subjected to models of focal ischemia, the nNOS mutant mice develop smaller infarcts, consistent with a role for nNOS in neurotoxicity following cerebral ischemia. In contrast, eNOS mutant mice develop larger infarcts, and show a more pronounced hemodynamic effect of vascular occlusion. The knockout mice also show that nNOS and eNOS isoforms differentially modulate the release of neurotransmitters in various regions of the brain. eNOS knockout mice respond to vessel injury with greater neointimal proliferation, confirming that reduced NO levels seen in endothelial dysfunction change the vessel response to injury. Furthermore, eNOS mutant mice still show a protective effect of female gender, indicating that the mechanism of this protection cannot be limited to upregulation of eNOS expression. The eNOS mutant mice also prove that eNOS modulates the cardiac contractile response to ß-adrenergic agonists and baseline diastolic relaxation. Atrial natriuretic peptide, upregulated in the hearts of eNOS mutant mice, normalizes cGMP levels and restores normal diastolic relaxation.