Functional and transcriptional induction of aquaporin-1 gene by hypoxia; analysis of promoter and role of Hif-1α.

Aquaporin-1 (AQP1) is a water channel that is highly expressed in tissues with rapid O(2) transport. It has been reported that this protein contributes to gas permeation (CO(2), NO and O(2)) through the plasma membrane. We show that hypoxia increases Aqp1 mRNA and protein levels in tissues, namely mouse brain and lung, and in cultured cells, the 9L glioma cell line. Stopped-flow light-scattering experiments confirmed an increase in the water permeability of 9L cells exposed to hypoxia, supporting the view that hypoxic Aqp1 up-regulation has a functional role. To investigate the molecular mechanisms underlying this regulatory process, transcriptional regulation was studied by transient transfections of mouse endothelial cells with a 1297 bp 5' proximal Aqp1 promoter-luciferase construct. Incubation in hypoxia produced a dose- and time-dependent induction of luciferase activity that was also obtained after treatments with hypoxia mimetics (DMOG and CoCl(2)) and by overexpressing stabilized mutated forms of HIF-1α. Single mutations or full deletions of the three putative HIF binding domains present in the Aqp1 promoter partially reduced its responsiveness to hypoxia, and transfection with Hif-1α siRNA decreased the in vitro hypoxia induction of Aqp1 mRNA and protein levels. Our results indicate that HIF-1α participates in the hypoxic induction of AQP1. However, we also demonstrate that the activation of Aqp1 promoter by hypoxia is complex and multifactorial and suggest that besides HIF-1α other transcription factors might contribute to this regulatory process. These data provide a conceptual framework to support future research on the involvement of AQP1 in a range of pathophysiological conditions, including edema, tumor growth, and respiratory diseases.

Fine structure of Henneguya hemiodopsis sp. n. (Myxozoa), a parasite of the gills of the Brazilian teleostean fish Hemiodopsis microlepes (Hemiodontidae)

A fish-infecting myxosporean, Henneguya hemiodopsis sp. n., found infecting the gills of Hemiodopsis microlepis and collected from the Poty River near the city of Teresina, Brazil, was described based on ultrastructural studies. The parasite occurred within large whitish polysporic plasmodia (up to 200 μm in diameter) containing asynchronous developmental sporogonic stages, mainly mature spores. The spores measured 19.7 ± 0.9 μm in total length (n = 30) and the ellipsoidal spore body was 10.8 ± 0.5 μm long, 3.3 ± 0.4 μm wide and 2.5 ± 0.5 μm thick. The spores were composed of two equal shell valves adhering together along the straight suture line, with each valve having equal-sized caudal tapering tails measuring 8.7 ± 0.6 μm in length. The spores were surrounded by a thin anastomosed network of microfibrils, more evident on the tails. There were two symmetric elongated bottle-like polar capsules 3.5 ± 0.3 μm long and 1.0 ± 0.2 μm wide, each with a polar filament with five to six coils. Given the morphological and ultrastructural differences from previously described parasites and the specificity of the host species, we propose a new species, named H. hemiodopsis sp. n.

Light and electron microscopy of Myxobolus sciades n. sp. (Myxozoa), a parasite of the gills of the Brazilian fish Sciades herzbergii (Block, 1794) (Teleostei: Ariidae)

A myxosporean parasite in the gill lamellae of the freshwater teleost fish, Sciades herzbergii (Ariidae) (Block, 1794), from the Poti River (Northeast of Brazil) was described by light and electron microscopy studies. Polysporic histozoic cyst-like plasmodia containing several life-cycle stages, including mature spores, were observed. The spores were pyriform and uninucleate, measuring 9.15 ± 0.39 μm (n = 50) long, 4.36 ± 0.23 μm (n = 25) wide and 2.61 ± 0.31 μm (n = 25) thick. Elongated pyriform polar capsules (PC) were of equal size (4.44 ± 0.41 μm long and 1.41 ± 0.42 μm in diameter) and each contained a polar filament with 9-10 coils obliquely arranged in relation to the axis of PC. The PC wall was composed of two layers of different electron densities. Histological analysis revealed the close contact of the cyst-like plasmodia with the basal portion of the epithelial gill layer, which exhibited some alterations in the capillary vessels. Based on the morphological and ultrastructural differences, the similarity of the spore features to those of the genus Myxobolus and the specificity of this host to previously described species, we describe a new species named Myxobolus sciades n. sp. in this study.

Molecular epidemiology of HIV-1 clades in Southern Brazil

Human immunodeficiency virus (HIV) clades B and C account for more than 60% of the HIV-1 infections worldwide. In this paper, we describe the profiles of patients infected with subtypes of HIV-1 from the state of Paraná, Southern Brazil, and correlate them with demographic and epidemiological findings. A retrospective analysis of HIV cases reported from 1999-2007 was also performed. Data from 293 patients were reviewed and 245 were older than 13 (58% female). The distribution of clades was as follows: B 140 (57%), C 67 (23%), F 24 (10%) and mosaic or unique recombinant forms (URFs) 24 (10%). Of the 48 patients younger than 13 years of age (62.5% male), vertical transmission occurred in 46 and the distribution of clades was as follows: B 14 (29%), C 24 (50%), F 7 (15%) and URFs 6 (13%). There was no significant difference in mortality between HIV-1 subtypes. In both groups, patients infected with clade C tended to have higher rates of injection drug use exposure risk.

Myxidium volitans sp. nov., a parasite of the gallbladder of the fish, Dactylopterus volitans (Teleostei: Triglidae) from the Brazilian Atlantic coast: morphology and pathology

Myxidium volitans sp. nov. (Myxozoa: Myxidiidae) parasitizing the hypertrophied green-brownish gallbladder of the teleost Dactylopterus volitans, collected in the Atlantic coast near Niterói, Brazil was described based on ultrastructural studies. The spores were fusiform, sometimes slightly crescent-shaped on average 21.7 ± 0.3 µm (mean ± standard deviation) (n = 50) long and 5.6 ± 0.4 µm (n = 30) wide. The spore wall was thin and smooth, comprising two equally-sized valves joined by a hardly visible sutural ridge. Spores containing two pyriform polar capsules (PC) (5.0 ± 0.4 × 2.3 ± 0.3 µm) (n = 30) are situated in each extremity of the spore. The PC wall was composed of hyaline layer (0.20-0.29 µm thick) and by a thin external granular layer. Each PC contains a polar filament (PF) with irregular arrangements that was projected from its apical region to the bases of PC and coiled laterally from bases to the tip of PC. Some regular striations and S-like structures in the periphery of the PFs with four-five irregular sections were observed. Based on the spore morphology, ultrastructural differences and the specificity of the host we describe this parasite as a new myxosporidian, named M. volitans sp. nov.

Targeting aquaporin function: potent inhibition of aquaglyceroporin-3 by a gold-based compound.

Aquaporins (AQPs) are membrane channels that conduct water and small solutes such as glycerol and are involved in many physiological functions. Aquaporin-based modulator drugs are predicted to be of broad potential utility in the treatment of several diseases. Until today few AQP inhibitors have been described as suitable candidates for clinical development. Here we report on the potent inhibition of AQP3 channels by gold(III) complexes screened on human red blood cells (hRBC) and AQP3-transfected PC12 cells by a stopped-flow method. Among the various metal compounds tested, Auphen is the most active on AQP3 (IC(50) = 0.8±0.08 µM in hRBC). Interestingly, the compound poorly affects the water permeability of AQP1. The mechanism of gold inhibition is related to the ability of Au(III) to interact with sulphydryls groups of proteins such as the thiolates of cysteine residues. Additional DFT and modeling studies on possible gold compound/AQP adducts provide a tentative description of the system at a molecular level. The mapping of the periplasmic surface of an homology model of human AQP3 evidenced the thiol group of Cys40 as a likely candidate for binding to gold(III) complexes. Moreover, the investigation of non-covalent binding of Au complexes by docking approaches revealed their preferential binding to AQP3 with respect to AQP1. The high selectivity and low concentration dependent inhibitory effect of Auphen (in the nanomolar range) together with its high water solubility makes the compound a suitable drug lead for future in vivo studies. These results may present novel metal-based scaffolds for AQP drug development.

Development and validation of PCR-based assays for diagnosis of American cutaneous leishmaniasis and identificatio nof the parasite species

In this study, PCR assays targeting different Leishmania heat-shock protein 70 gene (hsp70) regions, producing fragments ranging in size from 230-390 bp were developed and evaluated to determine their potential as a tool for the specific molecular diagnosis of cutaneous leishmaniasis (CL). A total of 70 Leishmania strains were analysed, including seven reference strains (RS) and 63 previously typed strains. Analysis of the RS indicated a specific region of 234 bp in the hsp70 gene as a valid target that was highly sensitive for detection of Leishmania species DNA with capacity of distinguishing all analyzed species, after polymerase chain reaction-restriction fragment length polymorfism (PCR-RFLP). This PCR assay was compared with other PCR targets used for the molecular diagnosis of leishmaniasis: hsp70 (1400-bp region), internal transcribed spacer (ITS)1 and glucose-6-phosphate dehydrogenase (G6pd). A good agreement among the methods was observed concerning the Leishmania species identification. Moreover, to evaluate the potential for molecular diagnosis, we compared the PCR targets hsp70-234 bp, ITS1, G6pd and mkDNA using a panel of 99 DNA samples from tissue fragments collected from patients with confirmed CL. Both PCR-hsp70-234 bp and PCR-ITS1 detected Leishmania DNA in more than 70% of the samples. However, using hsp70-234 bp PCR-RFLP, identification of all of the Leishmania species associated with CL in Brazil can be achieved employing a simpler and cheaper electrophoresis protocol.

Association analysis of human leukocyte antigen class II (DRB1) alleles with leprosy in individuals from São Luís, state of Maranhão, Brazil

Epidemiological studies have demonstrated that the variability of the clinical response to infection caused by Mycobacterium leprae is associated with host genetic factors. The present study investigated the frequency of human leukocyte antigen (HLA) class II (DRB1) alleles in patients with leprosy from São Luís, Maranhão, Brazil. A case-control study was performed in 85 individuals with leprosy and 85 healthy subjects. All samples were analysed via polymerase chain reaction-sequence specific oligonucleotide probes. The HLA-DRB1*16 allele showed a higher frequency in the group with leprosy [(9.41% vs. 4.12%) odds ratio (OR) = 2.41 95% confidence interval (CI) (0.96-6.08) p = 0.05], whereas the HLA-DRB1*11 allele was less frequent in the group with leprosy [(6.47% vs. 11.76%) OR = 0.51 95% CI (0.23-1.12) p = 0.09]. The frequency of HLA-DRB1* alleles between the control group and leprosy patient subgroups presenting different forms of the disease showed that the HLA-DRB1*16 (16.13% vs. 8.24%, OR = 4.10, CI = 1.27-13.27, p = 0.010) and HLA-DRB1*14 (5% vs. 3.53%, OR = 4.63, CI = 1.00-21.08, p = 0.032) alleles were significantly more frequent in patients with different clinical subtypes of leprosy. The sample size was a limitation in this study. Nevertheless, the results demonstrated the existence of a genetic susceptibility associated with the clinical forms of leprosy. The low frequency of the HLA-DRB1*11 allele should be further studied to investigate the possible protective effect of this allele.

Ultrastructural description of Ceratomyxa microlepis sp. nov. (Phylum Myxozoa): a parasite infecting the gall bladder of Hemiodus microlepis, a freshwater teleost from the Amazon River

A new ceratomyxid parasite was examined for taxonomic identification, upon being found infecting the gall bladder of Hemiodus microlepis (Teleostei: Hemiodontidae), a freshwater teleost collected from the Amazon River, Brazil. Light and transmission electron microscopy revealed elongated crescent-shaped spores constituted by two asymmetrical shell valves united along a straight sutural line, each possessing a lateral projection. The spores body measured 5.2 ± 0.4 µm (n = 25) in length and 35.5 ± 0.9 µm (n = 25) in total thickness. The lateral projections were asymmetric, one measuring 18.1 ± 0.5 µm (n = 25) in thickness and the other measuring 17.5 ± 0.5 µm (n = 25) in thickness. Two equal-sized subspherical polar capsules measuring 2.2 ± 0.3 µm in diameter were located at the same level, each possessing a polar filament with 5-6 coils. The sporoplasm was binucleate. Considering the morphometric data analyzed from the microscopic observations, as well as the host species and its geographical location, this paper describes a new myxosporean species, herein named Ceratomyxa microlepis sp. nov.; therefore representing the first description of a freshwater ceratomyxid from the South American region.

Detection of Mycobacterium leprae in saliva and the evaluation of oral sensitivity in patients with leprosy

The aim of this study was to investigate sensitivity disorders in the oral cavity related to the presence of Mycobacterium leprae in the saliva of treatment-naïve patients with leprosy in the state of Amazonas, Brazil. A cross-sectional study was conducted involving 45 subjects with leprosy. The subjects were interviewed to evaluate the sensitivity of the oral cavity. For the detection of M. leprae, saliva and slit-skin smear samples were collected. The samples were analysed using a bacteriological index (BI) protocol and the real-time quantitative polymerase chain reaction (qPCR). The results indicated that 15 of the 45 (33.3%) subjects with leprosy showed decreased oral sensitivity, which confirmed the importance of the oral cavity sensitivity evaluation. There was not a direct relationship between the presence of M. leprae in saliva and changes in oral sensitivity. Positive saliva qPCR results from six (31.6%) of 19 paucibacillary (PB) patients suggested the possibility of a new site for sample collection. Positive results using these diagnostic techniques (BI, slit-skin smear and saliva qPCR) increased to 55.5%, thus opening the possibility of combining these different techniques to increase the rate of positive diagnoses, especially in PB patients.

HLA and non-HLA genes in Behçet's disease: a multicentric study in the Spanish population.

INTRODUCTION According to genome wide association (GWA) studies as well as candidate gene approaches, Behçet's disease (BD) is associated with human leukocyte antigen (HLA)-A and HLA-B gene regions. The HLA-B51 has been consistently associated with the disease, but the role of other HLA class I molecules remains controversial. Recently, variants in non-HLA genes have also been associated with BD. The aims of this study were to further investigate the influence of the HLA region in BD and to explore the relationship with non-HLA genes recently described to be associated in other populations. METHODS This study included 304 BD patients and 313 ethnically matched controls. HLA-A and HLA-B low resolution typing was carried out by PCR-SSOP Luminex. Eleven tag single nucleotide polymorphisms (SNPs) located outside of the HLA-region, previously described associated with the disease in GWA studies and having a minor allele frequency in Caucasians greater than 0.15 were genotyped using TaqMan assays. Phenotypic and genotypic frequencies were estimated by direct counting and distributions were compared using the χ(2) test. RESULTS In addition to HLA-B*51, HLA-B*57 was found as a risk factor in BD, whereas, B*35 was found to be protective. Other HLA-A and B specificities were suggestive of association with the disease as risk (A*02 and A*24) or protective factors (A*03 and B*58). Regarding the non-HLA genes, the three SNPs located in IL23R and one of the SNPs in IL10 were found to be significantly associated with susceptibility to BD in our population. CONCLUSION Different HLA specificities are associated with Behçet's disease in addition to B*51. Other non-HLA genes, such as IL23R and IL-10, play a role in the susceptibility to the disease.

Indoor localisation with Android devices

This project analyses WiFiSLAM, an indoor positioning system for mobile phones that tries to estimate the position by analysing WiFi signals.

Educação continuada em enfermagem psiquiátrica: reflexão sobre conceitos

A preocupação com programas para capacitar o enfermeiro, para atuar em enfermagem psiquiátrica, levou-nos a este estudo, o primeiro da linha de pesquisa "Desenvolvimento do pessoal de enfermagem na área da psiquiatria e saúde mental". O objetivo deste foi fazer uma reflexão sobre os conceitos de educação continuada, em serviço e permanente existentes na literatura mais recente sobre o assunto. Para o levantamento bibliográfico dos conceitos em pauta, utilizamos as bases de dados MEDLINE e LILACS, dissertações e teses da área da enfermagem e alguns títulos clássicos sobre o assunto. Após análise dos conceitos e discussão com peritos, concluímos que a denominação educação contínua em serviço é a que guarda mais consonância com os autores e peritos consultados e a que mais atende às demandas de conhecimento existentes na área de assistência de enfermagem psiquiátrica rumo à qualidade da assistência.

Perfil do familiar cuidador de idoso fragilizado em convívio doméstico da grande Região do Porto, Portugal

Estudo descritivo com o objetivo de caracterizar o perfil do familiar cuidador, cujo método foi a aplicação dos instrumentos QPFC e WHOQOL-Bref da OMS a uma amostra de 120 familiares cuidadores de idosos dependentes, representantes da população referida de três Centros de Saúde da Região do Porto, Portugal. Os dados foram coletados durante o período de 01/2005 a 03/2005. Destacou-se, como cuidador principal, a mulher na idade em torno de 55 anos, que tem aos seus encargos, além do idoso, outros dependentes da família. As circunstâncias de cuidado, sejam pelas necessidades do idoso, ou requerimentos familiares, impunham conseqüências negativas à vida e à saúde. Contudo, ao mesmo tempo, esses cuidadores manifestavam suas percepções e sentimentos positivos da sensação confortadora, da dignificação de suas vidas, ao assumirem o papel de cuidador do idoso. Em conclusão, como enfermeiros, é essencial considerar o binômio cuidador e idoso dependente, pois exige atenção especial de cuidados da vida e saúde por parte dos serviços de saúde.

Habilidades de pensamento crítico no processo diagnóstico em enfermagem

O objetivo do estudo foi identificar habilidades de pensamento crítico no processo diagnóstico em enfermagem. Trata-se de um estudo exploratório descritivo com aplicação de caso clínico a sete discentes de enfermagem para identificação de habilidades de pensamento crítico e suas justificativas. Foi realizada análise de conteúdo para processamento dos dados. Tem-se que seis participantes citaram análise, conhecimento técnico-científico e raciocínio lógico como habilidades de pensamento crítico importantes na identificação de um diagnóstico de enfermagem prioritário; a experiência clínica foi citada por cinco participantes; o conhecimento sobre o paciente e aplicação de padrões foram habilidades mencionadas por três participantes; o discernimento e perspectiva contextual foram habilidades referidas por dois participantes. Com base nesses resultados, observou-se a utilização de habilidades de pensamento crítico relacionadas às etapas do processo de diagnóstico de enfermagem. Concluí-se que a aplicação desse processo configura uma estratégia que possibilita o desenvolvimento de habilidades específicas de pensamento crítico.