Role of microRNAs 221/222 on Statin Induced Nitric Oxide Release in Human Endothelial Cells

Background: Nitric oxide (NO) has been largely associated with cardiovascular protection through improvement of endothelial function. Recently, new evidence about modulation of NO release by microRNAs (miRs) has been reported, which could be involved with statin-dependent pleiotropic effects, including anti-inflammatory properties related to vascular endothelium function. Objective: To evaluate the effects of cholesterol-lowering drugs including the inhibitors of cholesterol synthesis, atorvastatin and simvastatin, and the inhibitor of cholesterol absorption ezetimibe on NO release, NOS3 mRNA expression and miRs potentially involved in NO bioavailability. Methods: Human umbilical vein endothelial cells (HUVEC) were exposed to atorvastatin, simvastatin or ezetimibe (0 to 5.0 μM). Cells were submitted to total RNA extraction and relative quantification of NOS3 mRNA and miRs -221, -222 and -1303 by qPCR. NO release was measured in supernatants by ozone-chemiluminescence. Results: Both statins increased NO levels and NOS3 mRNA expression but no influence was observed for ezetimibe treatment. Atorvastatin, simvastatin and ezetimibe down-regulated the expression of miR-221, whereas miR-222 was reduced only after the atorvastatin treatment. The magnitude of the reduction of miR-221 and miR-222 after treatment with statins correlated with the increment in NOS3 mRNA levels. No influence was observed on the miR-1303 expression after treatments. Conclusion: NO release in endothelial cells is increased by statins but not by the inhibitor of cholesterol absorption, ezetimibe. Our results provide new evidence about the participation of regulatory miRs 221/222 on NO release induction mediated by statins. Although ezetimibe did not modulate NO levels, the down-regulation of miR-221 could involve potential effects on endothelial function.

A new species of Leiostracus from Bahia, Brazil (Gastropoda, Pulmonata, Orthalicidae)

A remarkable new species of pulmonate snail was recently collected in a small Atlantic Rainforest fragment near the city of Canavieiras, state of Bahia, Brazil, an area known for a high diversity of land snails. It is described herein as Leiostracus fetidus sp. nov. and can be easily identified by its color pattern of irregular brown to black axial stripes on a white to yellow background, a reddish axial band "separating" the white peristome from the rest of the shell and a broad brown spiral band surrounding the umbilical region. Other diagnostic features include a relatively small size, a proto columellar fold and two very weak folds delimiting the basal region of the aperture. This discovery is a reminder of how little this fauna is known and also an alarm for proper conservation of these forest fragments.

Ensaios de micologia: contribuição dos cogumelos patogênicos em Minas Gerais

O autor diz que, de outubro de 1941 a dezembro de 1943, insulou numerosos fungos patogênicos de lesões as mais variadas. Obteve culturas puras de 5 casos de Rinocladiose, 3 de Moléstia de Lutz, apurou um Actinomyces minutissimus, 2 de Malassezia furfur e 2 Actinomyces, ainda em estudos, etc. Uma das observações de Esporotricose merece referência especial. Tratava-se de um indivíduo, morador em Governador Valadares, que apresentava uma lesão gomosa, nodular e verrucóide, fistulosa, localizada na região umbilical, provocada pela dentada de um peixe, de espécie ignorada, que atacou o paciente quando êste se banhava, completamente despido, nas águas do Rio Doce. O ponto da dentada sangrou ligeiramente e pouco depois começavam a aparecer os nódulos, que se sucederam progressivamente sem perturbar o estado geral do paciente. As sementeiras do pus de uma dessas gomas deram culturas puras do Rhinocladium Beurmanni, única espécie que o autor até hoje insulou, em 55 casos de Esporotricose em Minas Gerais. O autor já havia registrado a moléstia pela dentada de ratos, pelos ferimentos de espinhos de roseiras, pedaços de madeira, facas de cozinha, marteladas etc. Pensa, porém, que a moléstia produzida pela dentada de peixe é a primeira vez que se faz referência na literatura. Referiu-se depois o autor aos casos de Otomicoses que lhe chegaram às mãos em 1942, no serviço do Prof. ILDEU DUARTE. Dá o resumo das observações e concluiu que se tratava de lesões produzidas pelo Aspergillus fumigatus de FRESENIUS, 1841, de acôrdo com as lesões histopatológicas e as culturas que obteve.

Contribuição ao conhecimento das paralisias periféricas, múltiplas, uni-laterais, dos nervos craneanos (Síndrome de Villaret)

O autor descreve longamente uma observação dêste raro síndromo de lesão dos troncos nervosos periféricos dos nervos craneanos. No histórico do paciente, apurou a Micose pulmonar, pelo Neogeotrichum pulmoneum e úlcera estomacal, havendo-se curado de ambas. O síndromo nervoso atual foi de origem traumática, por arma de fogo, penetrando a bala no espaço retro-parotidiano e lesando, com maior ou menor intensidade, os nervos glossofaringeu, pneumogástrico, espinhal, hipoglosso e o simpatico cervical esquerdos. Além do síndromo nervoso, teve o paciente um aneurisma da artéria carótida primitiva, que foi ligado.

Toxoplasmose congênita

We had the opportunity to study 6 cases of the congenital form of toxoplasmosis, found in a series of 1200 necropsies of fetuses and newborn babies, realized at 3 different hospitals in Rio de Janeiro, Brazil. Among the 6 cases, 4 were premature babies liveborn at the 6th-8th gestational month and 2 were stillborn (1 premature and 1 at term). In all those cases, the diagnosis was based in the detection of the parasite in tissues and in one case it was even isolated the Toxoplasma from the necrotic material found in the cranial cavity. This strain of Toxoplasma, pathogenic to pigeons, to guinea pigs and to mice, is preserved by successive transfers in mice. Some facts observed in those cases present an interest not only strictly anatomic but also have certain value for the better acknowlegment of the disease. First, we want to call the attention to the presence of a sudden high fever, during or just before pregnancy in the 4 cases in which the maternal anamnesis was perfectly studied; this fever that was preceded by a normal beginning of pregnancy, had relatively rapid remission, but in 2 cases was immediately followed by uterine bleeding and premature delivery, although the puerperium had been apparently normal. It is known that are normal the subsequent children of the mothers that delivered a baby with toxoplasmosis and that several women have normal babies before the toxoplasmotic one. We believe that the fever observed in our cases could be indicative of the beginning of maternal infection and those are the reasons why we emphasize the need of careful anamnesis, specially in the cases actually diagnosed as inapparent infection. Another fact to notice is that in 5 of our cases the event premature delivery happened always between the 6th and the 8th months of pregnancy, and the only term fetus was delivered in advanced stage of maceration. The above mentioned facts could agree with the opinion of FRENKEL (1949), when he declared that "primary infection of the pregnant mother appears more likely to be the commoner mode of fetal toxoplasmic infection", but they would disagree with WEINMAN (1952) who believes that the transmission of Toxoplasma to the fetus is more frequent through a pregnant woman with chronic disease and who says "that infection contracted during pregnancy may and probably does happen from time to time"...Still in connection with the transmission of toxoplasmosis, we want to note the verification of inflammatory lesions in the placental villi and in the umbilical cord in 3 of the 4 cases in which such organs were examined at the microscope. In the case n. 1, we found several pseudocysts of Toxoplasma in the placenta, and the fibroblasts of Wharton's jelly were particularly rich in isolated forms and in colonies of Toxoplasma; the easy multiplication of the parasite in that tissue calls the attention and even suggests its utilisation for Toxoplasma's cultivation. The confirmation of Toxoplasma in human placenta was made only recently by CRISTEN et al. (1951) and by NEGHME et al. (1952), in Chile; it is not frequent in the literature, what gives some value to our present verification. Another observation was that provided by the case n. 6. This baby, a premature one of the 6th month, was 14 days old and-died with signs of respiratory disease, the causa mortis have been pneumonia. At the necropsy, we found no gross change that suggested toxoplasmosis, except the presence of some small necrotic focuses in the cerebral nervous substance around the ventricles. As a matter of fact, there was no enlargement of spleen or liver and neither leptomeningitis nor hydrocephalus. Such focuses were attributed to possible anoxia and in fact they are extremely similar to anoxial softenings, even when they are examined at the microscope; its structure composed of a central necrotic zone, surrounded by proliferated neuroglia and by a variable deposit of calcium salts, closely simulated the anoxial softenings, when the microscopical examination is based in the common histological preparations (hematoxilin-eosin, etc.). But when we examine preparations by the Giemsa or by the periodic acid-Schiff methods, we will note the presence of Toxoplasma, with its typical aspect or a little changed by degeneration. When we describe this observation, we wish to evidence the need of the search of Toxoplasma and closed parasites, in the cases of supposed pure anoxial softenings of nervous substance, in children. The frequency with which the congenital toxoplasmosis was anatomically verified should be emphasized, although the disease had not been clinically suspected, and it should be borne in mind that the second case of toxoplasmosis reported in the world was observed in Brazil by MAGARINOS TORRES; this case was the first to be described of the generalized congenital form of the infection, i. e. with myocardial lesions and parasites in skeletal muscles and skin.

Morbidade da esquistossomose mansoni no Brasil. I - Estudo de 4.652 casos observados no Rio de Janeiro de 1960 a 1979

No período de vinte anos (1960-1979), os autores observaram na Clínica de Doenças Infecciosas e Parasitárias da Faculdade de Medicina da Universidade Federal do Rio de Janeiro, sediada no Pavilhão Carlos Chagas do Hospital São Francisco de Assis, 4.652 casos de esquistossomose mansoni procedentes de 18 Estados do Brasil. Entre os casos observados 87,18% eram da forma hepatointestinal e 12,82% da forma hepatoesplênica entre os quais 30 tinham a forma pulmonar associada. Adicionalmente 30 casos da forma aguda ou toxémica foram estudados no mesmo Serviço, em famílias residentes no Rio de Janeiro. Além da observação clínica e do estudo epidemiológico incluindo o perfil migratório de cada caso e da casuística como um todo, foram realizados exames complementares como hemograma, mielograma, dosagem de transaminases, provas de função hepática, eletroforese de proteínas séricas, biópsia hepática, radiografias de tórax e de esôfago, esplenoportografia, eletrocardiograma e estudos hemodinâmicos do sistema porta, do coração e da artéria pulmonar em grupos de pacientes selecionados.

Angiostrongylus vasorum (Baillet, 1866) Nematoda: Prostostrongylidae, em cães de Minas Gerais, Brasil

Foi identificado Angiostrongylus vasorum (Baillet, 1866) colhido da artéria pulmonar de dois cães (Canis familiaris) procedentes do município de Caratinga, Estado de Minas Gerais, Brasil. É apresentada a descrição morfológicas do parasita. Esta é a primeira referência desse parasita no Estado de Minas Gerais.

Physa Marmorata Guilding, 1828 (Pulmonata: Physidae)

A description of Physa marmorata Guilding, 1828, based on material collected at its type-locality, the Caribbean island of Saint Vincent, is presented. The shell is thin, horn-colored, surface very glossy, diaphanous. Spire acute, elevated; protoconch distinct, rounded-conical, reddish-brown; five not shouldered, broadly convex whorls with subobsolete spiral lines and thin growth lines. Aperture elongated, 1.4-2.0 times as long as the remaining shell length, narrow obovate-lunate; upper half acute-angled,lower half oval,narrowly rounded at the base, outer lip sharp, inner lip completely closing the umbilical region; a very distinct callus on the parietal wall; columellar lip with a low ridge gradually merging into the callus. ratios: shell width/shell length = 0.44 - 0.52 (mean 0.47); spire length /shell lenght = 0.33-0.41 (mean 0.39); aperture length/shell lenght = 0.59-0.67 (mean 0.62). Oral lappets laterally mucronate, foot spatulate with deeply pigmented acuminate tail. Mantle reflection with 6-10 short triangular dentations covering nearly half the right surface of the body whorl, and 4-6 covering a part of the ventral wall. Body surface with tiny dots of greenish-yellow pigment besides melanin. Renal tube tightly folded in toa zigzag course. Ovotestis diverticula acinous, laterally pressed against each other around a collecting canal. Ovispermiduct with well-developed seminal vesicle. oviduct highly convoluted, merging into a less convoluted nidamental gland which narrows to a funnel-shaped uterus and a short vagina. Spermathecal body oblong, more or less constricted in the middle and somewhat curved; spermathecal duct uniformly narrow, a little longer than be body. About 20 prostatic diverticula, simple, bifurcate or divided into a few short branches, distalmost ones assembled into a cluster. Penis long, nearly uniformly narrow; penial canal with lateral opening about the junction of its middle and lower thirds. Penial sheath with a bulbous terminal expasion the tip of which isinserted into the caudal end of the prepuce. Prepuce shouldered, much wider than the narrow portion of the penial sheath. Penial sheath/prepuce ratio about 2.08 (1.45-2.75). The main extrinsic muscles of the penial complex are a retractor, with a branch attached to the bulb, and another to the caudal end of the penial sheath; and a protractor, with a branch attached to the shoulder of the prepuce and adjoining area of the penial sheath, and another to the caudal end of the penial sheath. Egg capsule C-shaped, with 10-30 elliptical eggs (snails 10mm long) measuring about 1.10 mm (0.90-1.32) through the long axis and surrounded by an inner and an outer lamellate membranes. Jaw a simple obtusely V-shaped plate. radula will be described separately.

Physa cubensis Pfeiffer, 1839 (Pulmonata: Physidae)

A description of Physa cubensis Pfeiffer, 1839, based on 15 speciments collected in Havana, Cuba, is presented. The shell, measuring 9.0 x 4,8mm to 12.3 x 6.4mm, is ovate-oblong, thin, diaphanous, horncolored, shining. Spire elevated, broadly conical; protoconch distinct, roundish, reddish-brown. About five moderately shouldered, roundly convex whorls, penultimate whorl expanded; spiral striation subobsolete; growth line faint on the intermediate whorls, clearly visible on the body whorl, crowded here and there. Suture well impressed. Aperture elongated 2.05 - 2.67 (mean 2.27) times as long as the remaining length of the shell, narrow obovulate-lunate; upper half acute-angled, lower half oval, narrowly rounded at the base; outer lip sharp, inner lip completely closing the umbilical region; a thick callus on the parietal wall; columellar plait well marked. Ratios: shell width/shell length - 0.52-0.61 (mean 0.55); spire length/shell length = 0.27 - 0.33 (mean 0.31); aperture length/shell length = 0.67 - 0.73 (mean 0.69). Oral lappets laterally mucronate; foot spatulate with acuminate tail. Mantle relection with 6 - 8 short triangular dentations in the right lobe (columellar side) and 4 - 6 in the left lobe (near the pneumostome). Renal tube tightly folded into a zigzag course. Ovotestis, ovispermiduct, seminal vesicle, oviduct, nidamental gland, uterus and vagina as in Physa marmorata (see Paraense, 1986, Mem. Inst. Oswaldo Cruz, 81: 459-469). Spermathecal body egg-shaped or pear-shaped; spermathecal ducta uniformly narrow with expanded base, a little longer than the body. Spermiduct, prostate and vas deferens as in P. marmorata (Paraense, loc. cit.). Penis wide proximally, narrowing gradually apicad; penial canal with subterminal outlet. Penial sheath following the width of the penis and ending up by a bulbous expansion somewhat narrower than the proximal portion. Penaial sheath/prepuce ration = 1,25 - 1,83 (mean 1.49). Prepuce much wider than the bulb of the penial shealth, moderately shouldered owing to the intromission of the bulb, and with a large gland in one side of its proximal half occupating about a third of its length. Extrinsic muscles of the penial complex as in P. marmorata. Jaw a simple obtusely V-shaped plate. Radula to be described separetely.

Age-specific Prevalence of Antibodies to Hepatitis A in Children and Adolescents from Rio de Janeiro, Brazil, 1978 and 1995: Relationship of Prevalence to Environmental Factors

The age-specific prevalence of antibodies to hepatitis A virus (anti-HAV) was determined in two different population groups with low socio-economic status from Rio de Janeiro city, Brazil, whose serum samples were collected 17 years apart (Population 1, 1978; Population 2, 1995). In Population 2, analysis of the anti-HAV prevalence was also carried out with respect to environmental factors. Population 1 was composed of 520 stored sera collected from the umbilical cord of term neonates and children aged 1 month to 6 years. In population 2, 720 serum samples were collected from children and adolescents with ages ranging from 1 to 23 years. The overall prevalence rate of anti-HAV in Population 1 and Population 2 was 65.6% and 32.1%, respectively. In Population 1, the anti-HAV prevalence reached 88% at the age of 3, while in Population 2, it increased from 4.5% in children under the age of 3 to 66% in the group of adolescents over the age of 14. The low exposure to HAV infection in younger children from Population 2 could be a result of improved environmental hygiene and sanitation, as demonstrated by the presence of piped water, waste and sewage disposal systems in most houses from this population group. These findings indicate a possible change in the prevalence of hepatitis A in Rio de Janeiro

Anionic Sites, Fucose Residues and Class I Human Leukocyte Antigen Fate During Interaction of Toxoplasma gondii with Endothelial Cells

Toxoplasma gondii invades and proliferates in human umbilical vein endothelial cells where it resides in a parasitophorous vacuole. In order to analyze which components of the endothelial cell plasma membrane are internalized and become part of the parasitophorous vacuole membrane, the culture of endothelial cells was labeled with cationized ferritin or UEA I lectin or anti Class I human leukocytte antigen (HLA) before or after infection with T. gondii. The results showed no cationized ferritin and UEA I lectin in any parasitophorous vacuole membrane, however, the Class I HLA molecule labeling was observed in some endocytic vacuoles containing parasite until 1 h of interaction with T. gondii. After 24 h parasite-host cell interaction, the labeling was absent on the vacuolar membrane, but presents only in small vesicles near parasitophorous vacuole. These results suggest the anionic site and fucose residues are excluded at the time of parasitophorous vacuole formation while Class I HLA molecules are present only on a minority of Toxoplasma-containig vacuoles.

Inflammatory response of endothelial cells to hepatitis C virus recombinant envelope glycoprotein 2 protein exposure

The hepatitis C virus (HCV) encodes approximately 10 different structural and non-structural proteins, including the envelope glycoprotein 2 (E2). HCV proteins, especially the envelope proteins, bind to cell receptors and can damage tissues. Endothelial inflammation is the most important determinant of fibrosis progression and, consequently, cirrhosis. The aim of this study was to evaluate and compare the inflammatory response of endothelial cells to two recombinant forms of the HCV E2 protein produced in different expression systems (Escherichia coli and Pichia pastoris). We observed the induction of cell death and the production of nitric oxide, hydrogen peroxide, interleukin-8 and vascular endothelial growth factor A in human umbilical vein endothelial cells (HUVECs) stimulated by the two recombinant E2 proteins. The E2-induced apoptosis of HUVECs was confirmed using the molecular marker PARP. The apoptosis rescue observed when the antioxidant N-acetylcysteine was used suggests that reactive oxygen species are involved in E2-induced apoptosis. We propose that these proteins are involved in the chronic inflammation caused by HCV.

Interferon-γ inhibits group B Streptococcus survival within human endothelial cells

Endothelial dysfunction is a major component of the pathophysiology of septicaemic group B Streptococcus (GBS) infections. Although cytokines have been shown to activate human umbilical vein endothelial cells (HUVECs), the capacity of interferon (IFN)-γ to enhance the microbicidal activity of HUVECs against GBS has not been studied. We report that the viability of intracellular bacteria was reduced in HUVECs activated by IFN-γ. Enhanced fusion of lysosomes with bacteria-containing vacuoles was observed by acid phosphatase and the colocalisation of Rab-5, Rab-7 and lysosomal-associated membrane protein-1 with GBS in IFN-γ-activated HUVECs. IFN-γ resulted in an enhancement of the phagosome maturation process in HUVECs, improving the capacity to control the intracellular survival of GBS.