Cardioplegia using low volumic cardioplegic agents: morphological study in isolated rabbit hearts

INTRODUÇÃO: A solução cardioplégica farmacológica busca eliminar as conseqüências do dano isquêmico, que é o resultado do desbalanço entre a oferta e o consumo de energia durante a parada dos batimentos cardíacos, nas cirurgias cardíacas com circulação extracorpórea. OBJETIVO: Este trabalho avalia experimentalmente as alterações estruturais e ultra-estruturais em coração isolado de coelhos submetidos à parada protegida pela Solução para Cardioplegia de Baixo Volume (SCBV). MÉTODO: O estudo compreendeu um grupo controle e dois grupos experimentais. No grupo I, a parada cardíaca foi obtida pela infusão da SCBV por 2 horas. No grupo II, o experimento foi conduzido da mesma forma até a parada protegida pela SCBV por duas horas, imediatamente procedeu-se à reperfusão com solução oxigenada de Ringer Locke (RL) por uma hora. No grupo controle os corações foram perfundidos com solução oxigenada de RL por duas horas. Após os experimentos, oito amostras de parede lateral do ventrículo esquerdo foram fixadas em formaldeído 10% e glutaraldeído 2,5% para análises histológica e ultra-estrutural. RESULTADOS: As células do miocárdio, fibroblastos e células endoteliais, observadas nos grupos experimentais I e II, apresentaram marginalização da heterocromatina, compactação do nucléolo, alteração na forma das mitocôndrias, compactação das cristas e aumento da densidade da matriz mitocondrial, indicando que tanto a estrutura nuclear quanto a das organelas citoplasmáticas foram alteradas em relação às células do grupo controle. CONCLUSÃO: As modificações estruturais foram decorrentes de uma adaptação fisiológica da célula, não sendo indicativas de oncose ou apoptose, sugerindo, portanto, que a solução cardioplégica utilizada foi eficiente para a preservação das células.

Matrix metalloproteinase (MMP)-2 and MMP-9 activity and localization during ventral prostate atrophy and regrowth

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Reduced allergic lung inflammation in rats following formaldehyde exposure: Long-term effects on multiple effector systems

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Polycation-Based Gene Therapy: Current Knowledge and New Perspectives

At present, gene transfection insufficient efficiency is a major drawback of non-viral gene therapy. The 2 main types of delivery systems deployed in gene therapy are based on viral or non-viral gene carriers. Several non-viral modalities can transfer foreign genetic material into the human body. To do so, polycation-based gene delivery methods must achieve sufficient efficiency in the transportation of therapeutic genes across various extracellular and intracellular barriers. These barriers include interactions with blood components, vascular endothelial cells and uptake by the reticuloendothelial system. Furthermore, the degradation of therapeutic DNA by serum nucleases is a potential obstacle for functional delivery to target cells. Cationic polymers constitute one of the most promising approaches to the use of viral vectors for gene therapy. A better understanding of the mechanisms by which DNA can escape from endosomes and traffic to enter the nucleus has triggered new strategies of synthesis and has revitalized research into new polycation-based systems. The objective of this review is to address the state of the art in gene therapy with synthetic and natural polycations and the latest advances to improve gene transfer efficiency in cells.

Anti-angiogenic effects of pterogynidine alkaloid isolated from Alchornea glandulosa

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Arterial diffuse intimal thickening associated with enzootic calcinosis of sheep

Foram feitos estudos morfométrico, imunoistoquímico e ultra-estrutural do espessamento intimal difuso (DIT) das artérias de 7 ovinos com sinais clínicos de calcinose enzoótica espontânea causada pela ingestão da planta Nierembergia veitchii. As lesões caracterizavam-se por deposição de sais de cálcio na média como placas e estrias que, com frequência faziam saliência para a luz, criando rugosidades e irregularidades da íntima. Não foram observadas calcificações na artéria pulmonar e no sistema venoso. Microscopicamente, as calcificações das artérias restringiam-se quase que exclusivamente à média. Na imunoistoquimica, foi verificada a-actina nas células da média e nas do espessamento intimal. Receptores para 1,25(OH)2D3 foram evidenciados nos núcleos das células musculares da média, da íntima e das células endoteliais. As análises morfométricas em microscopia ótica revelaram, nas artérias, DIT irregularmente distribuido sem relação com a intensidade dos processos de calcificação subjacente nem com a espessura da média remanescente. A morfometria das alterações ultra-estruturais das células musculares lisas da média e da íntima espessada, mostrou que nessas últimas foi verificado aumento de até 318% na fração volumétrica das organelas de síntese em detrimento dos elementos contráteis, quando comparados com os valores das células da média. Essas modificações indicam a transformação das células musculares lisas de um tipo contrátil para células de tipo sintético. Entre essses dois extremos foram evidenciadas diversas formas intermediárias. Foram observadas evidências histológicas e ultra-estruturais da transformação e migração de células da média através das fenestras da lâmina elástica interna, para a íntima, onde proliferam e formam o DIT. Foi concluído que o DIT é um componente constante nas lesões arterioscleróticas nas calcinoses dos ovinos induzidas pela planta N. veitchii e que as células predominantes são células musculares lisas provenientes de predecessoras da média. Foi constatado que o DIT só excepcionalmente apresenta calcificação, vascularização, presença de células espumosas e fendas de colesterol. Não foram vistas áreas de necrose. Sugere-se que o fator indutor das alterações arteriais é o 1,25(OH)2D3 contido em N. veitchii.

Synthetic and natural polycations for gene therapy: State of the art and new perspectives

Currently, the major drawback of gene therapy is the gene transfection rate. The two main types of vectors that. are used in gene therapy are based on viral or non-viral gene delivery systems. There are several non-viral systems that can be used to transfer foreign genetic material into the human body. In order to do so, the DNA to be transferred must escape the processes that affect the disposition of macromolecules. These processes include the interaction with blood components, vascular endothelial cells and uptake by the reticuloendothelial system. Furthermore, the degradation of therapeutic DNA by serum nucleases is also a potential obstacle for functional delivery to the target cell. Cationic polymers have a great potential for DNA complexation and may be useful as non-viral vectors for gene therapy applications. The objective of this review was to address the state of the art in gene therapy using synthetic and natural polycations and the latest strategies to improve the efficiency of gene transfer into the cell.

Heme-Dependent and Independent Soluble Guanylate Cyclase Activators and Vasodilation

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Biodegradable Nanoparticles Containing Benzopsoralens: An Attractive Strategy for Modifying Vascular Function in Pathological Skin Disorders

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

Choroidal vasculature in diabetic rats

This study aims to evaluate the diabetic influence on the choroidal vessels morphology. Twenty Wistar rats were divided into a control (CG) and a diabetic group (DG). The animals had the diabetes induced by an intra-venous injection of Alloxan (42 mg/kg). Transmission electron microscopy analysis focusing the choroidal vessels was done one (T2) and twelve (T3) months after the diabetes induction. The CG rats in T3 showed vesicles and dense bodies in the endothelial and pericytic cells; the same structures were observed in the DG at T2. The DG rats in T3 had even more and intense changes than the T2DG rats. The morphological evaluation indicates that the choroidal vessels are affected in diabetes and the disease accelerates degenerative processes in the rat choroidal vasculature.

Pathophysiology of mesenteric ischemia/reperfusion: a review.

During ischemia, the cell structures are progressively damaged, but restoration of the blood flow, paradoxically, intensifies the lesions caused by the ischemia. The mechanisms of ischemia injury and reperfusion (I/R) have not been completely defined and many studies have been realized in an attempt to find an ideal therapy for mesenteric I/R. The occlusion and reperfusion of the splanchnic arteries provokes local and systemic alterations principally derived from the release of cytotoxic substances and the interaction between neutrophils and endothelial cells. Substances involved in the process are discussed in the present review, like oxygen-derived free radicals, nitric oxide, transcription factors, complement system, serotonin and pancreatic proteases. The mechanisms of apoptosis, alterations in other organs, therapeutic and evaluation methods are also discussed.

Interação de Paracoccidioides brasiliensis com células endoteliais

Paracoccidioidomycosis has a variety of clinical manifestations and Paracoccidioides brasiliensis, the causative agent, may infect many tissues, most importantly the lungs. Migration of pathogenic yeasts to the endothelial cell layer is considered a prerequisite for multiple organ invasion and dissemination of the fungus. In this study of the adhesion of P. brasiliensis to endothelial cells in vitro, we investigated whether this adhesion could represent a mechanism of dissemination. To this end, as well as using conventional optical microscopy, an alternative in vivo technique was developed, to detect the presence of fungal cells in umbilical cords embedded in paraffin wax. An experiment on the migration of P. brasiliensis through an endothelial cell monolayer was carried out, and the migration of yeast cells was greater, and took less time, in control wells with no cells. The fungus crossed the monolayer, but, compared to control wells, the migration-rate was about 30% lower. This shows that the monolayer only partially blocked migration of the fungus. In these experiments, we had great difficulty finding P. brasiliensis adhered to the cell monolayer, when it was examined at different times, suggesting that migration of the fungus across the endothelial layer is very fast, and cannot normally be observed in cell culture in vitro. Thus, P. brasiliensis can cross the endothelium rapidly and probably invades deeper tissue.

Effects of pentoxifylline and n-acetylcysteine on injuries caused by ischemia and reperfusion of splanchnic organs in rats

Aim. Occlusion and reperfusion of splanchnic arteries cause local and systemic changes due to the release of cytotoxic substances and the interaction between neutrophils and endothelial cells. This study evaluated the role of pentoxifylline (PTX) and n-acetylcysteine (NAC) in the reduction of ischemia, reperfusion shock and associated intestinal injury. Methods. Sixty rats were divided into 6 groups of 10 animals. Rats in three groups underwent mesenteric ischemia for 30 minutes followed by 120 minutes of reperfusion, and were treated with saline (SAL-5 mL/kg/ h), pentoxifylline (PTX-50 mg/kg) or n-acetylcysteine (NAC-430 mg/kg/h). The other 3 groups underwent sham ischemia and reperfusion (I/R) and received the same treatments. Hemodynamic, biochemical and histological parameters were evaluated. Results. No significant hemodynamic or intestinal histological changes were seen in any sham group. No histological changes were found in the lung or liver of animals in the different groups. There was a progressive decrease in mean arterial blood pressure, from mean of 111.53 mmHg (30 minutes of ischemia) to 44.30±19.91 mmHg in SAL-I/R. 34.52±17.22 mmHg in PTX-I/R and 33.81±8.39 mmHg in NAC-I/R (P<0.05). In all I/R groups, there was a progressive decrease in: aortic blood flow, from median baseline of 19.00 mL/min to 2.50±5.25 mL/min in SAL-I/ R; 2.95±6.40 mL/min in PTX-I/R and 3.35±3.40 mL/min in NAC-I/R (P<0.05); in the heart rate, from mean baseline of 311.74 bpm to 233.33±83.88 bpm in SAL-I/R, 243.20±73.25 bpm in PTX-I/R and 244.92±76.05 bpm in NAC-I/R (P<0.05); and esophageal temperature, from mean baseline of 33.68°C to 30.53±2.05°C in SAL-I/R, 30.69±2.21°C in PTX-I/R and 31.43±1.03°C in NAC-I/R (P<0.05). In the other hand, there was an attenuation of mucosal damage in the small intestine of the animals receiving PTX, and only in the ileum of the animals receiving NAC. No changes were found in ileum or plasma malondialdehyde levels in any group. Conclusion. PTX was more efficient in reducing histological lesions than NAC, but neither treatment prevented hemodynamic changes during splanchnic organs I/R.

Effect of 6-months of physical exercise on the nitrate/nitrite levels in hypertensive postmenopausal women

Background: Evidences have showed that the incidence of arterial hypertension is greater in postmenopausal women as compared to premenopausal. Physical inactivity has been implicated as a major contributor to weight gain and abdominal obesity in postmenopausal women and the incidence of cardiovascular disease increases dramatically after menopause. Additionally, more women than men die each year of coronary heart disease and are twice as likely as men to die within the first year after a heart attack. A healthy lifestyle has been strongly associated with the regular physical activity and evidences have shown that physically active subjects have more longevity with reduction of morbidity and mortality. Nitric oxide (NO) produced by endothelial cells has been implicated in this beneficial effect with improvement of vascular relaxing and reduction in blood pressure in both laboratory animals and human. Although the effect of exercise training in the human cardiovascular system has been largely studied, the majority of these studies were predominantly conducted in men or young volunteers. Therefore, the aim of this work was to investigate the effects of 6 months of dynamic exercise training (ET) on blood pressure and plasma nitrate/nitrite concentration (NOx-) in hypertensive postmenopausal women. Methods: Eleven volunteers were submitted to the ET consisting in 3 days a week, each session of 60 minutes during 6 months at moderate intensity (50% of heart rate reserve). Anthropometric parameters, blood pressure, NOx- concentration were measured at initial time and after ET. Results: A significant reduction in both systolic and diastolic blood pressure values was seen after ET which was accompanied by markedly increase of NOx- levels (basal: 10 ± 0.9; ET: 16 ± 2 μM). Total cholesterol was significantly reduced (basal: 220 ± 38 and ET: 178 ± 22 mg/dl), whereas triglycerides levels were not modified after ET (basal: 141 ± 89 and ET: 147 ± 8 mg/dl). Conclusion: Our study shows that changing in lifestyle promotes reducftion of arterial pressure which was accompanied by increase in nitrite/nitrate concentration. Therefore, 6-months of exercise training are an important approach in management arterial hypertension and play a protective effect in postmenopausal women. © 2009 Zaros et al; licensee BioMed Central Ltd.

Intravascular lymphomatosis in the central nervous system of dogs: Immunohistochemical investigation in two cases

We report here two postmortem cases of dogs with intravascular lymphomatosis affecting the central nervous system. Intravascular lymphomatosis is represented by an exclusively intravascular proliferation of neoplastic lymphoid cells. To characterize the origin of the neoplastic cells, we have proceeded with immunohistochemical analysis to identify B and T lymphocytes and endothelial cells. The results showed predominance of cells from the T cell lineage, and no evidence of B cell origin was found. Few cells from one dog also exhibited cytoplasmatic staining for vimentin and Von Willebrand factor. Although in one case some immunophenotype diversity was observed, the massive presence of CD3 positive cells confirmed these neoplasms as intravascular lymphomatosis of T cell origin.