Endothelial dysfunction in DOCA-salt hypertension - Possible involvement of prostaglandin endoperoxides

The effects of the arachidonic acid metabolism inhibitors on the acetylcholine responses of aortae from control (CR) and deoxycorticosterone acetate (DOCA)-salt hypertensive (HR) rats were investigated. The acetylcholine decreased response observed in HR [relaxation (%): CR 95.5 +/- 2.7, n = 4; HR 52.0 +/- 6.3, n = 5, p < 0.05] was restored by the cyclooxygenase inhibitor piroxicam [relaxation (%): CR 99.8 +/- 0.2, n = 4; HR 86.0 +/- 4.0, n = 5] and by the thromboxane synthetase inhibitor and the thrombox ane A(2)/prostaglandin H-2 receptor antagonist ridogrel [relaxation (%): CR 92.1 +/- 4.4, n = 7; HR 93.1 +/- 2.0, n = 7] but not by the inhibitors of thromboxane synthetase, prostacyclin synthetase, cytochrome P-450 monooxygenase, and lipoxygenase. So, endoperoxide intermediates seem to be involved in the decreased endothelium-dependent relaxation to acetylcholine in DOCA-salt hypertension. (C) 1999 Elsevier B.V. All rights reserved.

Native crystal structure of a nitric oxide-releasing lectin from the seeds of Canavalia maritima

Here, we report the crystallographic study of a lectin from Canavalia maritima seeds (ConM) and its relaxant activity on vascular smooth muscle, to provide new insights into the understanding of structure/function relationships of this class of proteins. ConM was crystallized and its structure determined by standard molecular replacement techniques. The amino acid residues, previously suggested incorrectly by manual sequencing, have now been determined as I17, I53, S129, S134, G144, S164, P165, S187, V190, S169, T196, and S202. Analysis of the structure indicated a dimer in the asymmetric unit, two metal binding sites per monomer, and loops involved in the molecular oligomerization. These confer 98% similarity between ConM and other previously described lectins, derived from Canavalia ensiformis and Canavalia brasiliensis. Our functional data indicate that ConM exerts a concentration-dependent relaxant action on isolated aortic rings that probably occurs via an interaction with a specific lectin-binding site on the endothelium, resulting in a release of nitric oxide. (C) 2005 Elsevier B.V. All rights reserved.

Atypical beta-adrenoceptor subtypes mediate relaxations of rabbit corpus cavernosum

This study was performed to characterize the beta-adrenoceptor population in rabbit isolated corpus cavernosum (RbCC) by using nonselective and selective beta-adrenoceptor agonists and antagonists in functional assays. Metaproterenol, ritodrine, fenoterol, and 8-hydroxy-5-[(1R)-1-hydroxy-2-[N-[(1R)-2-(rho-methoxy-phenyl)1-methylethyl] amino] ethyl] carbostyril (TA 2005) (3-100 nmol each) dose dependently relaxed the RbCC preparations. These relaxations were markedly reduced by N-omega-nitro-L-arginine methyl ester (L-NAME; 10 muM) and 1H-[1,2,4]-oxadiazolo-[4,3,-a]quinoxalin-1-one (ODQ) (10 muM), whereas the adenylyl cyclase inhibitor SQ 22,536 [9-(2-tetrahydrofuryl)adenine] (10 muM) had no effect. In contrast, neither L-NAME nor ODQ affected the isoproterenol-induced RbCC relaxations, but SQ 22,536 abolished this response. Sildenafil (1 muM) significantly potentiated the relaxations induced by beta(2)-agonists without affecting the isoproterenol-evoked relaxations. Rolipram (10 muM) enhanced the relaxations elicited by isoproterenol but had no effect on those induced by the selective beta(2) agonists. Propranolol and (+/-)-1-[2,3-(dihydro-7-methyl-1H-inden-4-yl)oxy]-3-[(1-methylethyl)amino]-2-butanolhydrochloride (ICI 118,551) determined a rightward shift in the concentration-response curves to isoproterenol in a noncompetitive manner with a reduction of maximum response at the highest antagonist concentration, with the slope values significantly different from unity. Propranolol and ICI 118,551 had no effect on the relaxations elicited by fenoterol, TA 2005, metaproterenol, and ritodrine. Atenolol and 1-[2-((3-carbamoyl-4-hydroxy)phenoxy) ethylamino]-3-[4-(1-methyl-4-trifluoromethyl-2-imidazolyl)-phenoxy]2-propanol methanesulfonate (CGP 20712A) (0.1-10 muM) failed to affect the relaxations induced by all tested beta-adrenoceptor agonists. Our study revealed the existence of two atypical beta-adrenoceptors in the rabbit erectile tissue. Isoproterenol relaxes the rabbit cavernosal tissue by activating atypical beta-adrenoceptors coupled to adenylyl cyclase pathway, whereas the selective beta2-adrenoceptor agonists relax the RbCC tissue through another atypical beta-adrenoceptor subtype coupled to nitric oxide release from the sinusoidal endothelium.

Lidocaine effects on corneal endothelial cell ultrastructure

The purpose of this study was to determine whether intracameral commercial lidocaine 2% induces alterations on the rabbit corneal endothelium. Forty white rabbits received different substances inside the anterior chamber: group (G)1, no substance; G2 and G3 received lidocaine 2% with preservative in aqueous solution; G4 and G5, lidocaine 2% with preservative in gel solution; G6 and G7, the anesthetic preservative (metilparahydroxybenzoate 0.1%); and G8 and G9, lidocaine 2% without preservative in aqueous solution. The animals from G2, 4, 6 and 8 were sacrificed after 1 h, and from G3, 5, 7 and 9 after 24 h after injection of the substance inside the anterior chamber. The corneas were clinically evaluated and assessed by transmission and scanning electron microscopy. G1, 2, 6, 7, 8 and 9 animals had very similar characteristics in clinical, ultrastructural and morphometric evaluations; the G3 and G4 animals showed discrete edema and one animal in G5 had intense corneal edema. We conclude that lidocaine 2% with preservative induces few ultrastructural alterations in the corneal endothelial cells.

Lead poisoning in cattle grazing pasture contaminated by industrial waste

Lead poisoning is described in a herd of 120 Nelore cows of which 35 were affected. All affected cows died after a clinical course of 2-7 d with clinical signs related to cortical neurological disturbances. The source of Pb was fumes from a car battery recycling plant which has had a failure in its filtering system. Lead concentrations in liver and kidneys of 2 cows, in soil;and in grass where the cows were held were respective 39 to 431 ppm, 147 to 431 ppm,and 245ppm. No significant gross changes were observed. Histopathology revealed of neuronal necrosis, vacuolation of the neuropil and hypertrophy of the vascular endothelium inthecerebral cortices, degeneration of the epithelial cellsofrenal proximal tubules, and hemosiderosis of kidney, spleen and liver.

Corneal endothelial cell morphology of normal dogs in different ages

Endothelial cell function is essential to maintain corneal transparency, but unfortunately the regenerative capacity of the endothelium is limited. There are only a few reports describing the effect of age on morphologic appearance of corneal endothelial cells of dogs. Studies of normal corneal endothelial cells in humans and dogs have shown a decrease in endothelial cell density (ECD) and an increase in pleomorphism and polymegethism with advancing age. The purpose of this study was to investigate the effect of age on ECD and endothelial cell morphology in dogs. A total of 30 dogs were divided into three groups (10 dogs/group) based on age: group 1 (2-12 months old), group 2 (24-72 months old), and group 3 (84 months or older). Corneas were processed for light and scanning electron microscopy. Results showed only difference in cell density between group 1 and groups 2 and 3, showing an initial decrease in cell density as the animal matured. Whereas there was significantly greater variation in cell size within the dogs in group 3 than there was within the other two groups, suggesting that there was increased polymegethism and pleomorphism with advancing age.

Differential vascular adaptive response in spontaneously hypertensive and Wistar rats: Role of nitric oxide, and prehypertensive and hypertensive states

Stress-induced vascular adaptive response in SHR was investigated, focusing on the endothelium. Noradrenaline responses were studied in intact and denuded aortas from 6-week-old (prehypertensive) and 14-week-old (hypertensive) SHR and age-matched Wistar rats submitted or not to acute stress (20-min swimming and I-h immobilization 25 min apart), preceded or not by chronic stress (2 sessions 2 days apart of 1-h day immobilization for 5-consecutive days). Stress did not alter the reactivity of denuded aorta. Moreover, no alteration in the EC50 values was observed after stress exposure. In intact aortas, acute stress-induced hyporeactivity to noradrenaline similar between strains at both age. Chronic stress potentiated this adaptive response in 6- and 14-week-old Wistar but not in 6-week-old SHR, and did not alter the reactivity of 14-week-old SHR. Maximum response (g) in intact aortas [6-week-old: Wistar 3.25 +/- 0.12, Wistar/acute 1.95 +/- 0.12*, Wistar/chronic 1.36 +/- 0.21*(+), SHR 1.75 +/- 0.11, SHR/acute 0.88 +/- 0.08*, SHR/chronic 0.85 +/- 0.05*; 14-week-old: Wistar 3.83 +/- 0.13, Wistar/acute 2.72 +/- 0.13*, Wistar/chronic 1.91 +/- 0.19*', SHR 4.03 +/- 0.17, SHR/acute 2.26 +/- 0.12*, SHR/chronic 4.10 +/- 0.23; inside the same strain: *P < 0.05 relate to non-stressed rat, (+)P < 0.05 related to acute stressed rat; n = 6-18]. Independent of age and strain, L-NAME and endothelium removal abolished the stress-induced aorta hyporeactivity. Conclusion: the vascular adaptive response to stress is impaired in SHR, independently of the hypertensive state. Moreover, this vascular adaptive response is characterized by endothelial nitric oxide-system hyperactivity in both strains. (c) 2006 Elsevier B.V. All rights reserved.

Use of specular microscopy to determine corneal endothelial cell morphology and morphometry in enucleated cat eyes

The purpose of this study was to investigate the effect of age on endothelial morphology and morphometry in cats. The corneal endothelium was studied using a contact specular microscope. A total of 18 cats (Felis catus Linnaeus, 1758) were evaluated in this study. The subjects were divided into three groups of six cats each in function of age: G1 (1 to 3 months old), G2 (5 to 12 months old), and G3 (24 to 40 months old). The examination presented data as endothelial cell density (ECD), average cell area, corneal thickness, polymegathism, and pleomorphism. Results revealed ECD decrease in corneas of normal cats with age, as well as a corresponding increase in endothelial cell area and pleomorphism. The present work suggests that the endothelial parameters evaluated change with advancing age.

Morphological and Histochemical Characterization of Gill Filaments of the Brazilian Endemic Bivalve Diplodon expansus (Kuster, 1856) (Mollusca, Bivalvia, Hyriidae)

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

Microscopic studies on the transition between the sigmoid sinus, the superior bulb of the jugular vein and the first portion of the internal jugular vein

The author studied the structure of the tissue components of the tunicae of the terminal segment of the sigmoid sinus, particularly at the level of the transition between the sigmoid sinus, the superior bulb of the jugular vein and the first portion of the human internal jugular vein; it was established that the transition between the sigmoid sinus and the first portion of the internal jugular vein occupies the whole extension of the superior bulb of the jugular vein up to the inferior third of the first portion of this vessel. These vascular walls exhibit a structure similar to that of the dura, i.e. the tunica adventitia is formed by fascicles of collagenic fibers which describe discontinuous spirals, more open proximal to the beginning of the first portion of the internal jugular vein. Approximately in the inferior third of the first portion of the internal jugular vein, there appear fascicles of smooth muscle fibers which are arranged similarly to those of the venous walls. The tunica intima of these vascular segments exhibits an endothelium resting on a network of elastic fibers which may play the role of an internal elastic lamina. From the bony border of the jugular foramen there originates a connective system whose fascicles of collagenic and elastic fibers incorporate to the wall of the internal jugular vein after describing a stretch in spiral around the vascular lumen.

Papel da regiao AV3V no choque septico experimental em ratos tratados com NaCl 7,5%

The effect of intravenous infusion of hypertonic saline (HS) on the recovery of mean arterial pressure (MAP) during septic shock was studied in sham-operated rats and in rats with electrolytic lesion in the anteroventral third ventricle (AV3V) region. Our results show that intravenous HS infusion in rats treated with endotoxin (Etx) partially restores MAP, but when we have a severe shock produced by Etx, HS was not able to reverse the hypotension. We also show that the integrity of the AV3V region is essential for the protective action of HS in endotoxin shock. It is possible that NO production contributes to the deleterious effect of endotoxin. So, the unraveling of the release of NO by the vascular endothelium and their role as regulators of vascular tone is increasing our understanding of the physiology and pathophysiology of the cardiovascular system and will therefore enhance the possibilities of preventing and treating endotoxin shock.

Ultrastructural and morphometric aspects of ageing in the retinal capillaries of rats

Age-related morphological, ultrastructural and morphometric changes in the capillaries of the superficial and deep plexuses of the rat retina were studied in animals aged from 3 to 15 months. Our results suggest that age-related morphological alterations start occurring in the retina of rats at about 12 months of age. Increased glycogen deposits, pinocytotic vesicles, residual bodies and cell debris were observed in both the endothelial and pericytic cells of 12- and 15-month-old animals. In addition, heterogeneous osmiophilic accumulations, electron-transparent spaces were observed in the basement membrane as well as projections of the basement membrane towards the neighboring cells. Morphometric examination of the two vascular plexuses studied did not show differences in the area of the endothelial or pericytic cells, basement membrane or vascular lumen between rats of different ages.

Atherosclerosis morphology and pathogenesis

Atherosclerosis is a very common and important disease being the most important cause of mortality in Brazil. Indeed, in 1995, 23.3% of deaths, all ages, in our country, were the consequence of atherosclerosis. This percentage grows to 26.3% for S. Paulo and 32.7% for Rio Grande do Sul. Morphologically, there are 3 main types of lesions: fatty streaks, fibrous plaques, and complicated lesions. Fatty streaks are inocuous and occur early in life. In some persons, with age, they change into fibrous plaques that may lead to stenosis. They also may become complicated by erosion, calcification, hemorrhage and thrombosis. Atherosclerosis is initiated by endothelial functional alterations responsible for increase in permeability to macromolecules, adhesion, and migration of monocytes-macrophages and lymphocytes plus recruitment of platelets and smooth-muscle medial cells. Adhesion molecules, cytokines, growth factors, and free radicals are locally synthesized, favoring proliferation of extracellular matrix and progression of the lesion. Experimental, clinical, and epidemiological evidence point to the importance of lipids, mainly cholesterol-rich low-density lipoprotein (LDL), as one of the most important molecules involved in the genesis and progression of atherosclerosis. Patients with a genetic disorder of cholesterol metabolism (familial hyperlipidemia), caused by a decrease in the availability of receptors for LDL, develop severe atherosclerosis early in life. A series of other factors, such as age, diabetes melitus, diet, hypertension, lack of exercise, elevated hemocysteinemia, immunological disorders, and coagulation instability, are related to the progression of atherosclerosis. All of them are capable of altering the endothelium or increasing the offer of LDL. All the above-mentioned factors are systemic; but atherosclerosic lesions are focal, located at preferential sites such as the emergence of colaterals, bifurcations, and curvatures of arteries, all areas in which the laminar flow is disturbed. In these areas shear stress is diminished favoring the prolongation of permanence time of lipid particles, cells, cytokines, growth factors, etc., in the vicinity of the endothelium. Moreover, the endothelium has sensors that act as transducers of mechanical forces in biological responses. Experimental data demonstrate that the number and quality of adhesion molecules, cytokines, and growth factors synthetized, as well as the local production of radicals, and pro and anticoagulation factors may change with shear stress favoring or not the local establishment and progression of atherosclerotic lesions.

A complexa fisiopatologia dos episódios vasooclusivos na anemia falciforme

Hemolytic anemia and vasoocclusion are the cardinal clinical features of sickle cell anemia. Vasoocclusion is a complex process involving not only the polymerization of deoxygenated sickle hemoglobin tetramers, but also interactions between sickle erythrocytes, vascular endothelium, platelets, leukocytes, and plasma proteins. The increased adherence of sickle erythrocytes to endothelium has been implicated as an early step in vasoocclusion. Other researchers have focused on leukocytes and platelets which might also contribute to disturbed blood flow. Microvascular occlusion results in acute painful crises, whereas macrovascular occlusion seems to be the cause of organ failure. The anemia results from the markedly shortened circulatory survival of sickle erythrocytes, together with a limited erythropoietic response. The erythropoiesis increases intensively, but it is not enough to balance the increased rate of erythrocytes destruction to maintain normal levels of total erythrocytes and hemoglobin concentrations; mainly by the low oxygen affinity of hemoglobin S and increased 2,3-Diphosphoglycerate. It is very difficult to separate processes leading to anemia or to vasoocclusion. Understanding the involvement of multiple blood componentes in vasoocclusion may elucidate the clinical manifestations and complications of sickle cell anemia, and may give new insights into the preventive and curative therapy.