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.

Corneal endothelium of the Magellanic penguin (Spheniscus magellanicus) by scanning electron microscopy

The corneal endothelium is essential for the maintenance of the corneal transparency. The aim of this study was to examine the morphology of the endothelial surface and perform morphometric analysis of the normal corneal endothelial cells of the Magellanic penguin (Spheniscus magellanicus) using scanning electron microscopy. The present work demonstrates that the corneal endothelium of the Magellanic penguin is similar to those described in other vertebrates. Copyright 2005 by American Association of Zoo Veterinarians.

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.

Densidade das células do endotélio corneano em olhos de cães à microscopia especular

The aim of this study was to examine the endothelial surface and to perform a morphometric analysis of the corneal endothelial cells in normal eyes of dogs using specular microscopy. Morphometric analysis with regard mean cell area and cell density was performed. Both eyes of ten mixed-breed, males and females, with 6 years of age, weighing about 15 kg euthanatized for reasons unrelated to this study were evaluated. Eyes were examined to determine that they did not have visible ocular disease and transported to the laboratory in moist chamber. Using a contact specular microscope the corneal endothelium was examined. Three images of the central corneal endothelium of each eye were obtained. The mean cell area and the cell density of the corneal endothelial cells were obtained using software for corneal endothelium analysis and density measurement. The mean cell area was 395 ± 36 μm 2 and the endothelial cell density was 2555 ± 240 cells/mm2. The present work demonstrates that the normal corneal endothelium of dog is similar to those described in human.

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.

The expression of aquaporins 1 and 9 in adult rat epididymis is perturbed by chronic exposure to ethanol

Aquaporins (AQPs), notably AQP-1 and AQP-9, may contribute to reabsorption of fluid and solute across the epididymis. Ethanol is related to be a toxicant affecting directly or indirectly the epididymis and the sperm motility. This study examined the expression of AQP-1 and AQP-9 in adult epididymis of the UChA and UChB 10% (v/v) ethanol-preferring rats, focusing the ethanol-induced hormonal disturbances upon the regulation of these AQPs. Chronic ethanol intake significantly decreased body weight, while UChA and UChB rats displayed a marked loss of epididymal weights. Both ethanol-consuming animals had a severe reduction of testosterone levels, whereas LH and 17β-estradiol were unchanged. Throughout the epididymis, a strong reaction to AQP-1 was observed in myoid and endothelial cells of the UChB ethanol-preferring rats, differently from a moderate intensity in the initial segment of the UChA rats. In addition, AQP-9 showed a strong immunoreaction in the apical membrane of principal cells at initial segment. In cauda epididymis, the level of AQP-9 was reduced along the microvillus projections in both UChA and UChB rats compared to controls. We conclude that chronic ethanol consumption modulates the androgen levels, thereby modifying the expression pattern of AQP-1 and 9 in the epididymis. © 2011 Elsevier Ltd.

Cimetidine-induced vascular cell apoptosis impairs testicular microvasculature in adult rats

Cimetidine, an H2 receptor antagonist used for treatment of gastric ulcers, exerts antiandrogenic and antiangiogenic effects. In the testes cimetidine impairs spermatogenesis, Sertoli cells and peritubular tissue, inducing apoptosis in the myoid cells. Regarding the importance of histamine and androgens for vascular maintenance, the effect of cimetidine on the structural integrity of the testicular vasculature was evaluated. Adult male rats received cimetidine (CMTG) and saline (CG) for 50 days. The testes were fixed in buffered 4% formaldehyde and embedded in historesin and paraffin. In the PAS-stained sections, the microvascular density (MVD) and the vascular luminal area (VLA) were obtained. TUNEL method was performed for detection of cell death. Testicular fragments embedded in Araldite were analyzed under transmission electron microscopy. A significant decrease in the MVD and VLA and a high number of collapsed blood vessel profiles were observed in CMTG. Endothelial cells and vascular muscle cells were TUNEL-positive and showed ultrastructural features of apoptosis. These results indicate that cimetidine induces apoptosis in vascular cells, leading to testicular vascular atrophy. A possible antagonist effect of cimetidine on the H2 receptors and/or androgen receptors in the vascular cells may be responsible for the impairment of the testicular microvasculature.

Caracterização imunoistoquímica de neoplasias de bexigas associadas à hematúria enzoótica bovina

Bovine enzootic hematuria is characterized by the development of hemangiomatous lesions from several types of neoplastic processes, from epithelial and mesenchymal origin. In this research the histogenesis of neoplastic lesions found in bladder of bovines with enzootic hematuria from Caparaó microregion in the state of Espírito Santo, Brazil was determined. To accomplish this objective, immunohistochemical analysis was performed with primary antibodies: anti-vimentin, anti-cytokeratin, anti-CD31 and anti-uroplakin. Neoplasms found included urothelial carcinoma, in situ carcinoma, adenocarcinoma, hemangioma, myxoma e hemangiosarcoma. Immunohistochemical staining of cytokeratin in epithelial neoplasms and vimentin in mesenchymal neoplasms was significant (p<0.05). CD31 was positive in all the vessels of all samples, however, the staining was significant (p<0.05) in the tumor endothelial cells of the vascular mesenchymal neoplasms, as in hemangiomas and hemangiosarcomas. Uroplakin III staining was uneven in several neoplastic types and showed no significant difference (p>0.05). Most neoplasms showed an atypical uroplakin staining on urothelium and, in the case of hemangiosarcomas there was no staining of the urothelium at all. The Spearman statistical analysis revealed a positive correlation (r= 0.63, p= 0.05) between CD31 and vimentin and between cytokeratin and uroplakin (rs= -0.61, p= 0.05). It was concluded that biomarkers anti-cytokeratin, anti-vimentin and anti-CD31 are important for the diagnosis of neoplasms epithelial, mesenchymal and vascular mesenchymal, respectively. It is possible to use vimentin and CD31 in association in vascular mesenchymal neoplasms and cytokeratin and uroplakin in epithelial neoplasms. The uroplakin is an effective marker, not only for tumor diagnosis, but also to evaluate the urothelial integrity.

Facial hemangiossarcoma in a blue-fronted amazon parrot (Amazona aestiva)

Here we describe the first case report of an hemangiosarcoma in a male Blue-fronted amazon parrot (Amazona aestiva). The mass was localized near the inferior portion of the right orbit. A fine needle aspiration biopsy (FNAB) was performed and a presuntive diagnosis of hemangioma/hemangiosarcoma was established. Further histopathological evaluation revealed a mesenchymal neoplasm composed basically by well formed vascular channels lined by pleomorphic endothelial cells. Thus, the final diagnosis was a well-differentiated cutaneous hemangiosarcoma.