Serological evidence of Borrelia circulation among blood donors in the São Paulo state, Brazil.

OBJECTIVE: The objective of this study was to examine the Borrelia seroprevalence among blood donors in Southeast Brazil. BACKGROUND: There is evidence that Borrelia spirochetes are circulating in Brazil; however, there are no studies that characterise these bacteria and investigate their seroprevalence in the Brazilian population. Such a situation, combined with a recent outbreak of tick-borne Rocky Mountain spotted fever in the São Paulo state demonstrates the increasing role of ticks as arthropod vectors in Brazil. METHODS: For the purpose of the study, 452 blood donors from Ribeirão Preto city, São Paulo state were tested using anti-Borrelia immunoglobulin G (IgG) assay. The positive results were also confirmed by Western blot for anti-borrelia IgM/IgG. RESULTS: The anti-Borrelia IgG enzyme-linked immunosorbent assay (ELISA) showed nine positive and nine borderline reactive samples, giving a total seroprevalence of 2·0% of anti-Borrelia IgG among Brazilian blood donors. The confirmation of the seropositive and borderline samples by Borrelia Western blot was demonstrated by IgG-positive results in 16 samples (a seroprevalence of 3.5%). Anti-Borrelia IgM antibodies were also detected in one sample. CONCLUSIONS: Our results demonstrate that Borrelia-like spirochetes may be circulating among blood donors from the São Paulo State and that the ticks have an important epidemiological role as vectors of bacterial infections in this Brazilian region. These results not only alert us to possible actions that might be undertaken in order to completely characterise the aetiological agents of Lyme-like syndromes in Brazil but also the possible impact that these bacterial agents might have on haemotherapy practices.

Novel polymorphisms in the promoter region of the perforin gene among distinct Brazilian populations and their functional impact.

Cytotoxic T lymphocytes and natural killer cells play a crucial role in eliminating tumour and virus-infected cells. The perforin is a key part of the arsenal that these cells use to destroy their targets. In this study, we characterized single-nucleotide polymorphisms (SNPs) located in the promoter region of the perforin gene among distinct Brazilian ethnic groups. The study was carried out by sequencing this region in three groups: European, African and Asian descents. We demonstrated for the first time the occurrence of three new polymorphisms in the promoter region of gene PRF1: 494A/G (rs78058707), 720G/A (rs75925789) and 1176C/T (rs75183511). Three other SNPs already described in the literature 63A/G (rs35401316), 112A/G (rs10999428) and 1012C/T (rs35069510) were also detected. The SNPs are distributed differently in the ethnic groups studied. The 112G allele was observed at high frequency, especially among Asian descents (48.1%). The 1012T allele was detected only among European descents, the 494G allele only among Asian descents and 1176T allele only in African descents. Based on the association between the polymorphisms described, ten new haplotypes were originated. In functional analysis, we noticed that SNPs present in most common haplotypes cannot induce significant differences in expression levels of perforin alone. In conclusion, this study demonstrates for the first time the existence of three new polymorphisms in perforin promoter and, contrary to what was stated, the presence of these SNPs does not alter the levels of protein expression.

Evaluation of water quality in fee-fishing ponds located in Guarapiranga's sub basin, Upper Tietê River basin, São Paulo State.

This study intended to characterize the fee-fishing ponds located in Guarapiranga's sub-basin both in terms of structure and environmental conditions. The studied region was mapped and 15 out of 24 georeferenced fee-fishing ponds had their owners interviewed through semi-structured questionnaires. In addition to that, samples were collected from the seven fee-fishing ponds closest to the reservoir throughout a seasonal cycle divided in four sample collections carried out between November 2013 and August 2014, aiming at measuring the following variables in situ: water flow rates, pH, turbidity, electrical conductivity and dissolved oxygen concentration in affluents and effluents. Other samples were taken in order to assess total phosphorus (TP) and total nitrogen (TN) concentrations. Our results show a decrease in dissolved oxygen concentration along with an increase in TP and TN concentration in effluents, as compared to affluents. TP e TN concentrations exceeded the permitted values established by CONAMA 357/2005 - Class II resolution in all effluents at least during part of the year. High nutrient levels were observed in affluents, condition with which basins that have been affected by population growth are commonly met. It was verified, by interviewing the owners regarding venture management and by analyzing the collected samples, that there is significant correlation between the daily feeding and the TP and TN levels (p=0.021 and p=0.045, respectively), with the ponds where feeding is more intense having showed the highest nutrient levels in their effluents. The results highlight the importance of mitigation measures such as educating the owners on proper water management, the installation of pond aerators in conjunction with tanks the filters of which shall be composed of aquatic macrophytes in their effluents, as well as awareness campaigns aiming to reduce the usage of dough by fishermen in order to minimize detrimental impacts on Guarapiranga's sub-basin.

Human pegivirus-1 (HPgV-1) RNA prevalence and genotypes in volunteer blood donors from the Brazilian Amazon.

OBJECTIVES: The objectives of this study were to evaluate the prevalence of Human Pegivirus-1 (HPgV-1) viremia and genotype diversity among healthy blood donors from the Eastern Brazilian Amazon (city of Macapá, State of Amapá). There is little information for prevalence and circulation of HPgV-1 in this remote Brazilian region. MATERIALS AND METHODS: We conducted a study evaluating the HPgV-1 RNA prevalence and circulating genotypes in 431 volunteer blood donors originating from the Eastern Brazilian Amazon. The obtained HPgV-1 positive samples were submitted to sequencing and genotyping analysis in order to examine the genotype diversity of this virus in the Brazilian Amazon. RESULTS: Our results demonstrated a prevalence of HPgV-1 RNA in 9.5% of the tested blood donors. The phylogenetic analyses of the detected positive samples showed the presence of HPgV-1 genotypes 1, 2 and 3. The most frequently detected genotype was 2 (78.0% of the cases) represented by sub-genotypes 2A (39.0%) and 2B (39.0%). At lower rates, genotypes 1 (14.6%) and 3 (7.4%) were also detected. CONCLUSION: Our results revealed the presence of genotypes with European, Asiatic and African endemicity in Amazonian blood donors, probably due to the complex miscegenation processes that took place in this Brazilian region. More investigations, including information for the prevalence of HPgV-1 RNA in blood donors from other Latin American countries are needed to estimate the viremic rates and genotype distribution of this virus in a highly diverse continent like South America.

Functional expression of kinin B1 and B2 receptors in mouse abdominal aorta.

Previous studies have shown that the vascular reactivity of the mouse aorta differs substantially from that of the rat aorta in response to several agonists such as angiotensin II, endothelin-1 and isoproterenol. However, no information is available about the agonists bradykinin (BK) and DesArg(9)BK (DBK). Our aim was to determine the potential expression of kinin B(1) and B(2) receptors in the abdominal mouse aorta isolated from C57BL/6 mice. Contraction and relaxation responses to BK and DBK were investigated using isometric recordings. The kinins were unable to induce relaxation but concentration-contraction response curves were obtained by applying increasing concentrations of the agonists BK and DBK. These effects were blocked by the antagonists Icatibant and R-715, respectively. The potency (pD(2)) calculated from the curves was 7.0 +/- 0.1 for BK and 7.3 +/- 0.2 for DBK. The efficacy was 51 +/- 2% for BK and 30 +/- 1% for DBK when compared to 1 microM norepinephrine. The concentration-dependent responses of BK and DBK were markedly inhibited by the arachidonic acid inhibitor indomethacin (1 microM), suggesting a mediation by the cyclooxygenase pathway. These contractile responses were not potentiated in the presence of the NOS inhibitor L-NAME (1 mM) or endothelium-denuded aorta, indicating that the NO pathway is not involved. We conclude that the mouse aorta constitutively contains B(1) and B(2) subtypes of kinin receptors and that stimulation with BK and DBK induces contractile effect mediated by endothelium-independent vasoconstrictor prostanoids.

Analysis of food habits of skate Rioraja agassizii (Elasmobranchii, Rajidae) from southern Brazil.

Catches and exports of skate Rioraja agassizii place this species as "vulnerable to extinction" on the IUCN Red List; therefore, biological and ecological knowledge becomes an important instrument for its conservation control. This study described and quantified the diet composition of R. agassizii by means of stomach analysis contents in the periods 2005-2006 and 2012-2013. We analyzed and quantified stomach contents in terms of abundance (%N), weight (%M), frequency of occurrence (% FO), and index of relative importance (IRI). The results showed differences in the food rates between the periods. However, the groups of food items were the same: Teleostei fish, decapods, and mollusks. In 2005-2006, the diet consisted mainly of shrimp, however, in 2012-2013 it consisted of fish, followed by decapods, especially shrimps. The differences in diets may be attributed to shrimp abundance, which do not characterize a change in the eating habits in 2012-2013, because, in addition to fish, shrimps were also important food sources. The presence of a certain prey is more related to its availability rather than the feeding preference of skate. The amount of ingested items is associated to biological and environmental factors, so that further studies relating diet with capture area, seasonality, depth, and other factors should be conducted.

Evaluation of water quality in fee-fishing ponds located in Guarapirangas sub basin, Upper Tietê River basin, São Paulo State / Avaliação da qualidade da água de pesqueiros localizados na Sub-Bacia Guarapiranga, Alto Tietê, SP

Abstract This study intended to characterize the fee-fishing ponds located in Guarapiranga's sub-basin both in terms of structure and environmental conditions. The studied region was mapped and 15 out of 24 georeferenced fee-fishing ponds had their owners interviewed through semi-structured questionnaires. In addition to that, samples were collected from the seven fee-fishing ponds closest to the reservoir throughout a seasonal cycle divided in four sample collections carried out between November 2013 and August 2014, aiming at measuring the following variables in situ: water flow rates, pH, turbidity, electrical conductivity and dissolved oxygen concentration in affluents and effluents. Other samples were taken in order to assess total phosphorus (TP) and total nitrogen (TN) concentrations. Our results show a decrease in dissolved oxygen concentration along with an increase in TP and TN concentration in effluents, as compared to affluents. TP e TN concentrations exceeded the permitted values established by CONAMA 357/2005 - Class II resolution in all effluents at least during part of the year. High nutrient levels were observed in affluents, condition with which basins that have been affected by population growth are commonly met. It was verified, by interviewing the owners regarding venture management and by analyzing the collected samples, that there is significant correlation between the daily feeding and the TP and TN levels (p=0.021 and p=0.045, respectively), with the ponds where feeding is more intense having showed the highest nutrient levels in their effluents. The results highlight the importance of mitigation measures such as educating the owners on proper water management, the installation of pond aerators in conjunction with tanks the filters of which shall be composed of aquatic macrophytes in their effluents, as well as awareness campaigns aiming to reduce the usage of dough by fishermen in order to minimize detrimental impacts on Guarapiranga's sub-basin.

Resumo Este estudo objetivou caracterizar, em termos estruturais e ambientais pesqueiros localizados na sub-bacia da Guarapiranga. A região de estudo foi mapeada, e do total de 24 pesque-pague georreferenciados, 15 foram entrevistados com os proprietários através de questionários semi-estruturados e, nos sete pesque-pagues com tributários mais próximos da represa, foram coletadas amostras de água em um ciclo sazonal divididas em quatro coletas entre novembro de 2013 a agosto de 2014. Foram medidos in situ: vazão, pH, turbidez, condutividade elétrica e oxigênio dissolvido nos afluentes e efluentes, e também foram coletadas amostras de água para determinação da concentração de fósforo total (PT) e nitrogênio total (NT). Os resultados mostraram que há uma diminuição nas concentrações de oxigênio dissolvido e aumento nas concentrações de PT e NT nos efluentes em relação aos afluentes. Os efluentes apresentaram valores de PT e NT acima dos limites permitidos pela resolução CONAMA 357/2005 - Classe II, ao menos em um período do ano. Também foram identificados afluentes com cargas elevadas de nutrientes, característica de bacias impactadas pelo adensamento populacional. Através do questionário aplicado aos proprietários sobre o manejo dos empreendimentos e pelas análises de água verificou-se que houve correlação significativa entre o arraçoamento diário e as cargas de PT e de NT (p=0,021 e p=0,045, respectivamente), sendo que nos locais onde o arraçoamento foi mais intenso, a carga de nutrientes nos efluentes foi maior. Os resultados evidenciam a importância de medidas mitigadoras como orientações aos proprietários para o manejo adequado dos lagos, projetos de educação ambiental com os usuários para diminuir do uso de ceva para a captura dos peixes, além da instalação de aeradores artificiais nos lagos e tanques com filtros compostos de macrófitas aquáticas nos seus efluentes, minimizando os impactos na sub-bacia do Guarapiranga.

Binding affinities and activation of Asp712Ala and Cys100Ser mutated kinin B1 receptor forms suggest a bimodal scheme for the molecule of bound-DABK.

Mutant forms of kinin B(1) receptor (B(1)R) and analogs of the full agonist des-Arg(9)-bradykinin (DABK) were investigated aiming to verify the importance of selected receptor residues and of each agonist-peptide residue in the specific binding and activation. Linked by a specific disulfide bond (Cys(100)-Cys(650)), the N-terminal (N(t)) and the EC3 loop C-terminal (C(t)) segments of angiotensin II (AngII) receptor 1 (AT(1)R) have been identified to form an extracellular site for binding the agonist N(t) segment (Asp(1) and Arg(2) residues). Asp(712) residue at the receptor EC3 loop binds the peptide Arg(2) residue. By homology, a similar site might be considered for DABK binding to B(1)R since this receptor contains the same structural elements for composing the site in AT(1)R, namely the disulfide bond and the EC3 loop Asp(712) residue. DABK, Ala(n)-DABK analogs (n=Ala(1)-, Ala(2)-, Ala(3)-, Ala(4)-, Ala(5)-, Ala(6)-, Ala(7)-, Ala(8)-DABK), and other analogs were selected to binding wild-type, Asp712Ala and Cys100Ser mutated B(1)R receptors. The results obtained suggested that the same bimodal scheme adopted for AngII-AT(1)R system may be applied to DABK binding to B(1)R. The most crucial similarity in the two cases is that the N(t) segments of peptides equally bind to the homologous Asp(712) residue of both AT(1)R and B(1)R extracellular sites. Confirming this preliminary supposition, mutation of residues located at the B(1)R extracellular site as EC3 loop Asp(712) and Cys(100) caused the same modifications in biological assays observed in AT(1)R submitted to homologous mutations, such as significant weakening of agonist binding and reduction of post-receptor-activation processes. These findings provided enough support for defining a site that determines the specific binding of DABK to B(1)R receptors.

Overexpression of hsa-miR-125b during osteoblastic differentiation does not influence levels of Runx2, osteopontin, and ALPL gene expression.

Multipotent mesenchymal stromal cells (MSCs) were first isolated from bone marrow and then from various adult tissues including placenta, cord blood, deciduous teeth, and amniotic fluid. MSCs are defined or characterized by their ability to adhere to plastic, to express specific surface antigens, and to differentiate into osteogenic, chondrogenic, adipogenic, and myogenic lineages. Although the molecular mechanisms that control MSC proliferation and differentiation are not well understood, the involvement of microRNAs has been reported. In the present study, we investigated the role of miR-125b during osteoblastic differentiation in humans. We found that miR-125b increased during osteoblastic differentiation, as well as Runx2 and ALPL genes. To study whether the gain or loss of miR-125b function influenced osteoblastic differentiation, we transfected MSCs with pre-miR-125b or anti-miR-125b and cultured the transfected cells in an osteoblastic differentiation medium. After transfection, no change was observed in osteoblastic differentiation, and Runx2, OPN, and ALPL gene expression were not changed. These results suggest that the gain or loss of miR-125b function does not influence levels of Runx2, OPN, and ALPL during osteoblastic differentiation.

Projeto saúde mental brincando em família: caracterizando sociodemograficamente crianças cardiopatas hospitalizadas e familiares / Mental health project toying in family: sociodemographic characterization of hospitalized cardiopathy children and of relatives

Objetivo: caracterizar o perfil sociodemográfico e de morbidade da clientela assistida, bem como os fatores associados com a qualidade do atendimento psicológico realizado entre janeiro e dezembro de 2013. Métodos: trata-se de um estudo exploratório, com 357 crianças e seus familiares, no Hospital Ana Nery em Salvador, Bahia, Brasil. Resultados: a maioria das crianças era do sexo masculino, pardas ou negras e com idade média de cinco anos. Verificou-se que aspectos relativos aos familiares, assim como o local onde ocorreram as sessões, influenciaram na qualidade das interações estabelecidas entre familiares e acolhedores do projeto. Conclusões: essa forma de cuidar demonstra a importância de incluir aspectos psicossociais capazes de ampliar as estratégias de cuidado para além do enfoque clínico, contribuindo para promoção e reabilitação em saúde mental.

Objective: characterize the sociodemographic and morbidity profile of the assisted clientele as wellas the factors associated with the quality of psychological care performed between January and December 2013. Methods: we conducted an exploratory study with 357 children and their families at the Ana Nery Hospital in Salvador, Bahia, Brazil. Results: we observed that them a majority of children were male, mixed race or black, and the average age was five years old. Family related aspects and the location in which these sessions took place influenced the quality of interactions established between family members and Play Therapists. Conclusions: this way of caring demonstrates the importance of incorporating psycho-social aspects to extend care strategies beyondthe clinical focus, contributing to the promotion and rehabilitation in mental health.

Evaluation of water quality in fee-fishing ponds located in Guarapirangas sub basin, Upper Tietê River basin, São Paulo State

Abstract This study intended to characterize the fee-fishing ponds located in Guarapiranga's sub-basin both in terms of structure and environmental conditions. The studied region was mapped and 15 out of 24 georeferenced fee-fishing ponds had their owners interviewed through semi-structured questionnaires. In addition to that, samples were collected from the seven fee-fishing ponds closest to the reservoir throughout a seasonal cycle divided in four sample collections carried out between November 2013 and August 2014, aiming at measuring the following variables in situ: water flow rates, pH, turbidity, electrical conductivity and dissolved oxygen concentration in affluents and effluents. Other samples were taken in order to assess total phosphorus (TP) and total nitrogen (TN) concentrations. Our results show a decrease in dissolved oxygen concentration along with an increase in TP and TN concentration in effluents, as compared to affluents. TP e TN concentrations exceeded the permitted values established by CONAMA 357/2005 Class II resolution in all effluents at least during part of the year. High nutrient levels were observed in affluents, condition with which basins that have been affected by population growth are commonly met. It was verified, by interviewing the owners regarding venture management and by analyzing the collected samples, that there is significant correlation between the daily feeding and the TP and TN levels (p=0.021 and p=0.045, respectively), with the ponds where feeding is more intense having showed the highest nutrient levels in their effluents. The results highlight the importance of mitigation measures such as educating the owners on proper water management, the installation of pond aerators in conjunction with tanks the filters of which shall be composed of aquatic macrophytes in their effluents, as well as awareness campaigns aiming to reduce the usage of dough by fishermen in order to minimize detrimental impacts on Guarapirangas sub-basin.

Resumo Este estudo objetivou caracterizar, em termos estruturais e ambientais pesqueiros localizados na sub-bacia da Guarapiranga. A região de estudo foi mapeada, e do total de 24 pesque-pague georreferenciados, 15 foram entrevistados com os proprietários através de questionários semi-estruturados e, nos sete pesque-pagues com tributários mais próximos da represa, foram coletadas amostras de água em um ciclo sazonal divididas em quatro coletas entre novembro de 2013 a agosto de 2014. Foram medidos in situ: vazão, pH, turbidez, condutividade elétrica e oxigênio dissolvido nos afluentes e efluentes, e também foram coletadas amostras de água para determinação da concentração de fósforo total (PT) e nitrogênio total (NT). Os resultados mostraram que há uma diminuição nas concentrações de oxigênio dissolvido e aumento nas concentrações de PT e NT nos efluentes em relação aos afluentes. Os efluentes apresentaram valores de PT e NT acima dos limites permitidos pela resolução CONAMA 357/2005 Classe II, ao menos em um período do ano. Também foram identificados afluentes com cargas elevadas de nutrientes, característica de bacias impactadas pelo adensamento populacional. Através do questionário aplicado aos proprietários sobre o manejo dos empreendimentos e pelas análises de água verificou-se que houve correlação significativa entre o arraçoamento diário e as cargas de PT e de NT (p=0,021 e p=0,045, respectivamente), sendo que nos locais onde o arraçoamento foi mais intenso, a carga de nutrientes nos efluentes foi maior. Os resultados evidenciam a importância de medidas mitigadoras como orientações aos proprietários para o manejo adequado dos lagos, projetos de educação ambiental com os usuários para diminuir do uso de ceva para a captura dos peixes, além da instalação de aeradores artificiais nos lagos e tanques com filtros compostos de macrófitas aquáticas nos seus efluentes, minimizando os impactos na sub-bacia do Guarapiranga.

Distribution of human immunodeficiency virus type 1 subtypes in the State of Amazonas, Brazil, and subtype C identification.

Few studies have reported the molecular epidemiological characterization of HIV-1 in the Northern region of Brazil. The present study reports the molecular and epidemiological characterization of 31 HIV-1 isolates from blood donors from the State of Amazonas who donated blood between April 2006 and March 2007. Serum/plasma samples from all donors were screened for HIV antibodies by ELISA and the results confirmed by Western blot analysis. Genomic DNA was extracted from the buffy coat using the Super Quik-Gene-DNA Isolation kit. Nested PCR was performed on the env, gag, and pol regions of HIV-1 using the Gene Amp PCR System 9700. Sequencing reactions were performed using the inner PCR primers and the DYEnamic™ ET Dye Terminator Kit, and phylogenetic analysis was performed using the gag, pol, and env gene sequences. We collected samples from 31 blood donors who tested positive for HIV-1 in confirmatory experiments. The male:female ratio of blood donors was 3.4:1, and the mean age was 32.4 years (range: 19 to 61 years). Phylogenetic analysis showed that subtype B is the most prevalent among Northern Brazilian HIV-1-seropositive blood donors. One HIV-1 subtype C and one circulating recombinant form (CRF_BF) of HIV-1 were identified in the State of Amazonas. This is the first study showing the occurrence of a possible "homogenous" subtype C in this region of Brazil. This finding could contribute to a better characterization of the HIV-1 strains that circulate in the country.

Analysis of food habits of skate Rioraja agassizii (Elasmobranchii, Rajidae) from southern Brazil / Análise quantitativa dos hábitos alimentares da raia Rioraja agassizii (Elasmobranchii, Rajidae) do Sudeste e Sul do Brasil

Abstract Catches and exports of skate Rioraja agassizii place this species as “vulnerable to extinction” on the IUCN Red List; therefore, biological and ecological knowledge becomes an important instrument for its conservation control. This study described and quantified the diet composition of R. agassizii by means of stomach analysis contents in the periods 2005-2006 and 2012-2013. We analyzed and quantified stomach contents in terms of abundance (%N), weight (%M), frequency of occurrence (% FO), and index of relative importance (IRI). The results showed differences in the food rates between the periods. However, the groups of food items were the same: Teleostei fish, decapods, and mollusks. In 2005-2006, the diet consisted mainly of shrimp, however, in 2012-2013 it consisted of fish, followed by decapods, especially shrimps. The differences in diets may be attributed to shrimp abundance, which do not characterize a change in the eating habits in 2012-2013, because, in addition to fish, shrimps were also important food sources. The presence of a certain prey is more related to its availability rather than the feeding preference of skate. The amount of ingested items is associated to biological and environmental factors, so that further studies relating diet with capture area, seasonality, depth, and other factors should be conducted.

Resumo A raia-santa, Rioraja agassizii é uma espécie endêmica da qual pouco se conhece sobre sua biologia e ecologia, o que a leva a ser classificada como “vulnerável a extinção”. O objetivo foi identificar e quantificar a composição da dieta alimentar da R. agassizii através da análise do conteúdo estomacal de exemplares capturados nos períodos de 2005-2006 e 2012-2013. Foram analisados os conteúdos alimentares e quantificados em abundância (N%), peso (M%), frequência de ocorrência (FO%) e índice de importância relativa (IRI) de cada item. Os resultados mostraram que em 2012-13 a dieta foi composta por peixes, seguido de decápodes, especialmente camarões, entretanto em 2005-06 foi composta basicamente por camarões. A diferença nas dietas não quer dizer que os hábitos alimentares mudaram, pois em 2012-13 além dos peixes, os camarões também foram classificados como maior importância alimentar. Considerando que a quantidade e variedade dos itens ingeridos está relacionada, principalmente, a fatores ambientais são necessários mais estudos que relacionem áreas de captura, sazonalidade e profundidade.

Overexpression of hsa-miR-125b during osteoblastic differentiation does not influence levels of Runx2, osteopontin, and ALPL gene expression

Multipotent mesenchymal stromal cells (MSCs) were first isolated from bone marrow and then from various adult tissues including placenta, cord blood, deciduous teeth, and amniotic fluid. MSCs are defined or characterized by their ability to adhere to plastic, to express specific surface antigens, and to differentiate into osteogenic, chondrogenic, adipogenic, and myogenic lineages. Although the molecular mechanisms that control MSC proliferation and differentiation are not well understood, the involvement of microRNAs has been reported. In the present study, we investigated the role of miR-125b during osteoblastic differentiation in humans. We found that miR-125b increased during osteoblastic differentiation, as well as Runx2 and ALPL genes. To study whether the gain or loss of miR-125b function influenced osteoblastic differentiation, we transfected MSCs with pre-miR-125b or anti-miR-125b and cultured the transfected cells in an osteoblastic differentiation medium. After transfection, no change was observed in osteoblastic differentiation, and Runx2, OPN, and ALPL gene expression were not changed. These results suggest that the gain or loss of miR-125b function does not influence levels of Runx2, OPN, and ALPL during osteoblastic differentiation.

Distribution of human immunodeficiency virus type 1 subtypes in the state of Amazonas, Brazil, and subtype C identification

Few studies have reported the molecular epidemiological characterization of HIV-1 in the Northern region of Brazil. The present study reports the molecular and epidemiological characterization of 31 HIV-1 isolates from blood donors from the State of Amazonas who donated blood between April 2006 and March 2007. Serum/plasma samples from all donors were screened for HIV antibodies by ELISA and the results confirmed by Western blot analysis. Genomic DNA was extracted from the buffy coat using the Super Quik-Gene-DNA Isolation kit. Nested PCR was performed on the env, gag, and pol regions of HIV-1 using the Gene Amp PCR System 9700. Sequencing reactions were performed using the inner PCR primers and the DYEnamic™ ET Dye Terminator Kit, and phylogenetic analysis was performed using the gag, pol, and env gene sequences. We collected samples from 31 blood donors who tested positive for HIV-1 in confirmatory experiments. The male:female ratio of blood donors was 3.4:1, and the mean age was 32.4 years (range: 19 to 61 years). Phylogenetic analysis showed that subtype B is the most prevalent among Northern Brazilian HIV-1-seropositive blood donors. One HIV-1 subtype C and one circulating recombinant form (CRF_BF) of HIV-1 were identified in the State of Amazonas. This is the first study showing the occurrence of a possible "homogenous" subtype C in this region of Brazil. This finding could contribute to a better characterization of the HIV-1 strains that circulate in the country.

Functional expression of kinin B1 and B2 receptors in mouse abdominal aorta

Previous studies have shown that the vascular reactivity of the mouse aorta differs substantially from that of the rat aorta in response to several agonists such as angiotensin II, endothelin-1 and isoproterenol. However, no information is available about the agonists bradykinin (BK) and DesArg9BK (DBK). Our aim was to determine the potential expression of kinin B1 and B2 receptors in the abdominal mouse aorta isolated from C57BL/6 mice. Contraction and relaxation responses to BK and DBK were investigated using isometric recordings. The kinins were unable to induce relaxation but concentration-contraction response curves were obtained by applying increasing concentrations of the agonists BK and DBK. These effects were blocked by the antagonists Icatibant and R-715, respectively. The potency (pD2) calculated from the curves was 7.0 ± 0.1 for BK and 7.3 ± 0.2 for DBK. The efficacy was 51 ± 2 percent for BK and 30 ± 1 percent for DBK when compared to 1 æM norepinephrine. The concentration-dependent responses of BK and DBK were markedly inhibited by the arachidonic acid inhibitor indomethacin (1 æM), suggesting a mediation by the cyclooxygenase pathway. These contractile responses were not potentiated in the presence of the NOS inhibitor L-NAME (1 mM) or endothelium-denuded aorta, indicating that the NO pathway is not involved. We conclude that the mouse aorta constitutively contains B1 and B2 subtypes of kinin receptors and that stimulation with BK and DBK induces contractile effect mediated by endothelium-independent vasoconstrictor prostanoids.