Diversity of fungi associated with macroalgae from an estuarine environment and description of Cladosporium rubrum sp. nov. and Hypoxylon aveirense sp. nov.

Fungal communities associated with macroalgae remain largely unexplored. To characterize algicolous fungal communities using culture dependent methods, macroalgae were collected from different sampling sites in the Ria de Aveiro estuary, Portugal. From a collection of 486 isolates that were obtained, 213 representative isolates were selected through microsatellite-primed PCR (MSP-PCR) fingerprinting analysis. The collection yielded 33 different genera, which were identified using the ITS region of the rDNA. The results revealed that the most abundant taxa in all collections were Acremonium-like species: Alternaria, Cladosporium, Leptobacillium and Penicillium. The fungal community composition varied with macroalgae species. Through multilocus phylogenetic analyses based on ITS, tub2, tef1-α and actA sequences, in addition to detailed morphological data, we propose Cladosporium rubrum sp. nov. (type strain=CMG 28=MUM 19.39) and Hypoxylon aveirense sp. nov. (type strain=CMG 29=MUM 19.40) as novel species.

Description of Freudenthalidium gen. nov. and Halluxium gen. nov. to Formally Recognize Clades Fr3 and H as Genera in the Family Symbiodiniaceae (Dinophyceae).

The Symbiodiniaceae are a family of marine dinoflagellates known mostly for their endosymbiotic interactions with invertebrates and protists, but facultatively and exclusively free-living life histories in this family are also evident. A recent systematic revision of the Symbiodiniaceae replaced the clade-based nomenclature of seven divergent lineages of "Symbiodinium" sensu lato with one based on formally described genera. The revised taxonomy was not extended to the whole group because type species to describe a new genus for each of the remaining clades and subclades were lacking. In an effort to characterize benthic habitats of symbiodiniaceans in sediments at Heron Island (Great Barrier Reef, Australia), we isolated >100 monoclonal Symbiodiniaceae cultures. Four of these belonged to Symbiodiniaceae 'subclade' Fr3, and three to Clade H, based on nucleotide sequence similarity (ITS2, LSU, cp23S, and mtCOB), representing the first cultures of these taxa. Based on these isolates, we propose two new genera: Freudenthalidium gen. nov. and Halluxium gen. nov., circumscribing Clades Fr3 and H, respectively. Three new species are described: Freudenthalidium heronense, F. endolithicum, and Halluxium pauxillum. Kofoidian tabulations of motile cells confirm previous observations that amphiesmal vesicle arrangements are generally conserved across the family. These descriptions are an important step toward completing the systematic revision of the Symbiodiniaceae. That this contribution was enabled by isolates from an endopsammic habitat highlights the potential of discovering new symbiodiniacean species in the environment, the study of which will lead to a deeper understanding of free-living versus symbiotic life histories in this ecologically important family of dinoflagellates.

Symbiolite formation: a powerful in vitro model to untangle the role of bacterial communities in the photosynthesis-induced formation of microbialites.

Microbially induced calcification is an ancient, community-driven mineralisation process that produces different types of microbialites. Symbiolites are photosynthesis-induced microbialites, formed by calcifying co-cultures of dinoflagellates from the family Symbiodiniaceae and bacteria. Symbiolites encase the calcifying community as endolithic cells, pointing at an autoendolithic niche of symbiotic dinoflagellates, and provide a rare opportunity to study the role of bacteria in bacterial-algal calcification, as symbiodiniacean cultures display either distinct symbiolite-producing (SP) or non-symbiolite-producing (NP) phenotypes. Using Illumina sequencing, we found that the bacterial communities of SP and NP cultures differed significantly in the relative abundance of 23 genera, 14 families, and 2 phyla. SP cultures were rich in biofilm digesters from the phylum Planctomycetes and their predicted metagenomes were enriched in orthologs related to biofilm formation. In contrast, NP cultures were dominated by biofilm digesters from the Bacteroidetes, and were inferred as enriched in proteases and nucleases. Functional assays confirmed the potential of co-cultures and bacterial isolates to produce biofilms and point at acidic polysaccharides as key stimulators for mineral precipitation. Hence, bacteria appear to influence symbiolite formation primarily through their biofilm-producing and modifying activity and we anticipate that symbiolite formation, as a low-complexity in vitro model, will significantly advance our understanding of photosynthesis-induced microbial calcification processes.

Cuttlefish capsule: An effective shield against contaminants in the wild.

Increasing anthropogenic pressures in estuaries are responsible for the rise of contaminants in several compartments of these ecosystems. Species that benefit from the nursery services provided by estuaries are exposed to such contaminants (e.g. metals and metalloids). It is therefore relevant to understand if marine invertebrates that use these areas as spawning grounds accumulate contaminants in their tissues throughout embryogenesis. This study aimed to quantify As, Co, Cr, Cu, Mn, Ni, Se, Pb, V and Zn concentrations in both capsule and embryos of the common cuttlefish (Sepia officinalis) in Sado Estuary (Portugal). Moreover, embryos at their initial, intermediate and final stage of development were collected in sites subjected to different anthropogenic pressures. In general, the capsule accumulated higher element concentration throughout embryogenesis which indicates that the capsule acts as an effective barrier against contaminants uptake by the embryo. Although the capsule becomes thinner throughout embryogenesis, embryo's protection does not seem to be compromised at later development stages. Additionally, the higher concentrations of As, Cu, Se and Zn in the embryo in comparison to the capsule suggests important biological roles during the embryogenesis of this cephalopod mollusc.

Lymphocyte subset reference intervals in blood donors from northeastern Brazil.

UNLABELLED: The reference intervals for leukocytes and lymphocytes currently used by most clinical laboratories present limitations as they are primarily derived from individuals of North American and European origin. The objective this study was to determine reference values for peripheral blood B lymphocytes, T lymphocyte subsets (CD4+, CD8+, naïve, memory, regulatory, TCRαß and TCRγδ+) and NK cells from blood donors in Salvador-Bahia, Brazil. RESULTS: The proportion of included male subjects was 73.7% and the median ages of males (34) and females (35) were found to be similar. Absolute counts total lymphocytes subsets to both gender was 1,956 (1,060-4,186) cells and relative values 34%. The T CD4+ and T CD8+ lymphocytes relative values was 51% (20-62) and 24% (9-28), respectively. The most statistically significant finding observed was a higher percentage of B lymphocytes (p=0.03) in females. Commonly cited subset reference intervals were found to be consistent with values in several populations from different geographic areas.

Dengue virus 4 (DENV-4) re-emerges after 30 years in Brazil: cocirculation of DENV-2, DENV-3, and DENV-4 in Bahia.

Dengue fever (DF) is a mosquito-borne viral disease of great concern in tropical and subtropical regions of the world. One important cause of the increase in DF is rapid development and urbanization has led to proliferation of the Aedes aegypti mosquito, the vector responsible for transmission of the illness. Surveillance of dengue virus (DENV) infection in Brazil shows the predominance of DENV-1, DENV-2, and DENV-3 until 2010. This study reports the reappearance of DENV-4 in Brazil for the first time in 30 years. Serum samples were collected from individuals (n = 214) exhibiting fever and muscular pain in Bahia, Brazil, during 2011-2012. These samples were subjected to reverse transcription-polymerase chain reaction (RT-PCR)/nested PCR, which revealed that 82% of samples were positive for DENV-4; most were older age groups and exhibited a serological pattern consistent with a primary infection. The cocirculation of multiple DENV serotypes within the same city places the population at risk for a fatal form of the disease. Therefore, with the increasing incidence of severe DF cases, early diagnosis will be a priority for public health efforts in Brazil.

Bioprospecting of marine invertebrates for new natural products - a chemical and zoogeographical perspective.

Bioprospecting for new marine natural products (NPs) has increased significantly over the last decades, leading to an unprecedented discovery of new molecules. Marine invertebrates have been the most important source of these NPs, with researchers commonly targeting particular taxonomic groups, marine regions and/or molecules from specific chemical groups. The present review focuses on new NPs identified from marine invertebrates between 2000 and 2009, and performs a detailed analysis on: (1) the chemical groups of these NPs; (2) the association of particular chemical groups to specific marine invertebrate taxa; and (3) the yielding of molecules from the same chemical group from organisms occurring in a particular geographic region. Our survey revealed an increasing number of new terpenoids being discovered between 2000 and 2009, contrasting with the decreasing trend in the discovery of new alkaloids and aliphatic molecules. Overall, no particular association was identified between marine invertebrate taxa and chemical groups of new NPs. Nonetheless, it is worth noting that most NPs recorded from cnidarians and mollusks were terpenoids, while most NPs identified in echinoderms were aliphatic compounds or carbohydrates. The geographical trends observed in our study do not support the idea of particular chemical groups of new NPs being associated with marine invertebrates from any specific geographical region, as NPs from different chemical groups were commonly distributed worldwide.

Alloimmunization and clinical profile of sickle cell disease patients from Salvador-Brazil.

Sickle cell disease (SCD) is an important public health issue in Bahia, Brazil. Erythrocyte transfusions may reduce morbidity of SCD, however, they are associated with numerous risks. Among other risk categories, alloimmunization to red cell antigens may result from transfusions. The aim of this study was to compare the clinical profile of transfused adult SCD patients with and without alloantibodies. The study included 108 patients (105 homozygous SS and three with hemoglobinopathy SC), followed in the Outpatient Unit of the Hematology and Hemotherapy Center of Bahia. A retrospective review of clinical records of adult SCD patients who received at least three red blood cell transfusions from 2004 to 2007 was performed. Transfusion units were phenotypically matched for ABH-D and C,c,E,e, and K antigens. Alloimmunization developed in 56 patients (53 SS and three SC). The most prevalent alloantibodies were anti-E, anti-K, and anti-C (39.3%, 21.4%, and 16.1%, respectively). Age, sex and positive antiglobulin test displayed statistically significant differences. Prevalence of clinical complications such as leg ulcers, stroke, and others did not show differences between groups. In conclusion, alloimmunization did not significantly modify the clinical outcomes of SCD patients from Bahia, Brazil.

Ancestry informative markers and complete blood count parameters in Brazilian blood donors / Marcadores informativos de ancestralidade e parâmetros no hemograma de doadores de sangue brasileiros

A complete blood count is very useful in clinical diagnoses when reference ranges are well established for the population. Complete blood counts and allele frequencies of Ancestry Informative Markers (AIMs) were analyzed in Brazilians with the aim of characterizing the hematological values of an admixed population. Positive associations were observed between gender and neutrophils, monocytes, eosinophils, erythrocytes, hemoglobin, hematocrit, MCV, MCHC and platelet counts. No significant differences were found for age, alcohol consumption, educational status, ethnicity, smoking in respect to the complete blood count values. In general, men had higher red blood cell values, while women had higher values for white blood cells and platelets. The study of the population was highly heterogeneous with mean proportions (± SE) of African, European and Amerindian ancestry being 49.0 ± 3.0 percent, 44.0 ± 9.0 percent and 7.0 ± 9.0 percent, respectively. Amerindian ancestry showed limited contribution to the makeup of the population, but estimated ancestral proportions were statistically significant (r = 0.9838; P<0.001). These hematologic values are similar to Afro-Americans, another admixed population.

O hemograma é muito útil no diagnóstico quando o intervalo de referência é adequadamente estabelecido para população. Com o objetivo de verificar os valores hematológicos em população heterogênea foi analisado o hemograma e frequências alélica de marcadores informativos de ancestralidade de brasileiros. Foi observada associação positiva entre sexo e os valores de neutrófilos, monócitos, eosinófilos, eritrócitos, hemoglobina, hematócrito, MCV, MCHC e plaquetas (IC 95 por cento; P<0,05). E não houve diferenças entre idade, consumo de álcool, nível educacional, etnia, tabagismo e os valores do hemograma (IC 95 por cento; P>0,05). Os homens apresentaram valores maiores no eritrograma, enquanto no leucograma e plaquetograma foram as mulheres. Foi observado também que a população é altamente heterogênea e as médias proporcionais (±DP) de ancestralidade Africana, Europeia e Ameríndia estimada foram: 49,0 ± 3,0 por cento, 44,0 ± 9,0 por cento e 7,0 ± 9,0 por cento, respectivamente. A contribuição ancestral ameríndia se demonstrou pequena, mas a estimativa de proporções ancestrais foi estatisticamente significante (r = 0,9838; P<0,001). Os valores hematológicos aqui descritos são parecidos com os descritos em negros americanos, outra população heterogênea.

Evaluation of factors that can modify platelet-rich plasma properties.

OBJECTIVE: The aim of this article is to discuss a protocol for obtaining platelet-rich plasma (PRP) and evaluate which factors, derived from its preparation method and from whole blood, modify PRP cytometry and coagulation time. STUDY DESIGN: Whole blood, harvested from 50 rabbits, was centrifuged at 300g for 10 minutes. Supernatant was recentrifuged at 5000g for 5 minutes. PRP was clotted with calcium chloride. Whole blood and PRP cytometry were obtained through automatic measurement. The amount of erythrocyte- and platelet-poor plasma drawn from whole blood was measured. Hematocrit, platelet and leukocyte count, mean corpuscular volume (MCV) and mean platelet volume (MPV), mean, standard deviation, and median were also calculated at whole blood and PRP. PRP coagulation time was also analyzed. Mean values between groups were analyzed using Student t test. Correlations were evaluated using Pearson's correlation coefficient. The significance level was set at P < .05. A linear regression was performed to investigate the relationship among the correlated variables. RESULTS: From whole blood, 2.68 mL of erythrocytes and 5.72 mL of platelet-poor plasma (PPP) were removed. PRP platelet count was 2,324,080 cells/microL. Whole blood hematocrit influenced the amount of cells and PPP removed, as well as PRP platelet count. PRP platelet count was dependent on whole blood hematocrit and platelet count, and does not interfere in PRP coagulation time. A linear interaction was confirmed between the variables that presented significant Pearson correlation. CONCLUSIONS: The protocol evaluated produces a good PRP. Whole-blood parameters can predict PRP features. Whole-blood hematocrit is an important variable for PRP preparation and PRP cytometry characterization. PRP platelet count is dependent upon whole-blood platelet count.

Prevalência de porphyromonas gingivalis. e leucometria, em mestiços brasileiros com periodontite crónica severa / Porphyromonas gingivalis prevalence and leukometry in Brazilians of mixed race with severe chronic periodontitis

Objetívos: Sendo a periodontite uma doença multifatorial que se inicia e é mantida pela agressão das bactérias periodontopatogênicas, este trabalho objetivou identificar a presença de Pgingvalis (Pg) no biofilme subgengival em indivíduos com periodontite crónica severa, bem como avaliar o leucograma respectivo.Material e métodos: Oitenta e quatro voluntários não-fumantes foram selecionados para o estudo: 43 pacientes com periodontite crónica severa (grupo caso - PCS) e 41 indivíduos sem periodontite (grupo controle - NP). Parâmetros clínicos periodontais e o leucograma foram avaliados. Amostras do biofilme subgengival foram coletadas para detecção de Pg pela técnica da reação em cadeia da polimerase (PCR). Para a análise estatística foram usados os testes t-student e Mann-Whitney (p<0.05). Resultados: Foi detectada a presença de Pg no biofilme subgengival de 29 pacientes (67,4%) do grupo PCS, não sendo observado este periodontopatógeno em nenhum dos indivíduos do grupo NP O grupo PCS apresentou significantemente valores mais elevados do que o NP no número de leucócitos totais e no número absoluto de neutrófilos. Conclusões: Os resultados mostraram que na maioria dos pacientes do grupo PCS foi detectada a presença de Porphyromonas gingivalis e que esta infecção bacteriana pode refletir-se na leucometria.

Anticorpos antifosfolípides em mulheres com antecedentes de perdas gestacionais: estudo caso-controle / Antiphospholipid antibodies in women with recurrent pregnancy loss: a case-control study

OBJETIVO: determinar a prevalência de anticorpos antifosfolípides em mulheres com antecedentes de perdas gestacionais na populacão obstétrica em geral e verificar se os anticorpos antifosfolípides representam fator de risco para perdas gestacionais na populacão estudada. MÉTODOS: foi realizado um estudo caso-controle prospectivo com mulheres grávidas e não grávidas, atendidas numa maternidade pública entre marco de 2003 e junho de 2004. As mulheres foram divididas em dois grupos de acordo com o passado obstétrico; 100 mulheres com antecedentes de perdas gestacionais de acordo com a definicão estabelecida para o diagnóstico da síndrome antifosfolipídica e que não apresentassem outros fatores relacionados ao insucesso gestacional; 150 mulheres saudáveis com antecedentes de duas ou mais gestacões bem sucedidas. A determinacão do anticoagulante lúpico (AL) foi feita mediante os testes de TTPA, dRVVT de triagem e dRVVT confirmatório. Para a pesquisa dos anticorpos anticardiolipina (aCL) classes IgG e IgM foi utilizado o teste de ELISA. e os resultados semiquantitativos expressos em unidades GPL e MPL. RESULTADOS: o anticoagulante lúpico estava presente em 5 por cento das pacientes-caso e 2 por cento dos controles (p=0,27). Os anticorpos aCL IgG estavam presentes em 18 por cento das pacientes-caso e 8,7 por cento das pacientes-controle (p=0,028; OR=2,3; IC 95 por cento=1-53). Na classe IgM, 5 por cento de positividade para os casos e 1 por cento para os controles (p=0,21). CONCLUSÕES: os anticorpos antifosfolípides (AL e/ou aCL IgG e/ou IgM) foram mais prevalentes nas mulheres com perdas gestacionais (28 por cento) que na populacão obstétrica em geral (17 por cento). As mulheres com aCL IgG têm duas vezes mais chance de ter perdas gestacionais que a populacão obstétrica em geral.

Educação física: a mediação na perspectiva bioética / Physical Education - Mediation on the Perspectives of Bioethics

O presente trabalho tem como objetivo refletir sobre a existência de discriminação velada em aulas de Educação Física. Foi desenvolvido a partir de pesquisa de campo realizada em uma escola pública do Distrito Federal, tendo como base os fundamentos da Bioética. Aborda a Educação Física diante dos princípios teóricos e avalia sua prática sob perspectivas bioéticas. Essa abordagem permitiu constatar uma dicotomia conceitual dos termos da expressão educação física e a avaliação, a existência de eufemismos mascarando a discriminação. Na prática, enquanto a dicotomia leva à descaracterização do processo educativo, pelo não-atendimento a seus pressupostos, o eufemismo responde por situações de exclusão durante as aulas, corroborando com possíveis segregações sociais. Os métodos adotados na escola, a mediação do professor e as necessidades do pré-adolescente na fase crítica de sua formação podem justificar o processo de interiorização experimentado e manifestado por ele em suas respostas. O problema pode ser observado, do ponto de vista pedagógico, com uma retomada crítica dos referidos pressupostos, conceitos e métodos e, bioeticamente, mediante a visão do outro como alteridade.

A discriminaçäo nas aulas de Educaçäo Física sob o enfoque bioético: um estudo de caso no Distrito Federal / Discrimination in the Physical Education classes under a bioethical approach: a case study in the Distrito Federal - Brazil

O presente estudo foi desenvolvido com o objetivo de identificar, descrever e analisar a ocorrência de procedimentos discriminatórios nas aulas de Educaçäo Física em uma escola da rede pública do Distrito Federal. Realizou-se uma revisäo de literatura abordando conceitos, legislaçäo e aspectos culturais da prática dsportiva na escola. Fundamentou-se o trabalho em um dos princípios da bioética: a alteridade. A pesquisa de campo desenvolvida mediante técnicas de observaçäo e entrevista, envolveu duas categorias de sujeitos: professores e alunos de 5ª a 8ª séries do Ensino Fundamental e foi desenvolvida por meio de técnicas de observaçäo e entrevista. A análise dos resultados possibilitou a identificaçäo da presença sutil da discriminaçäo nas atitudes dos sujeitos. Pôde-se inferir que os alunos, socialmente desfavorecidos, näo estariam tendo acesso a oportunidades de superaçäo de sua condiçäo desprivilegiada, em funçäo da existência de tratamento diferenciado entre alunos talentosos e näo-talentosos.

Frequência dos anticorpos antifosfolípides (aFL) em portadores de lúpus eritematoso sistêmico (LES) no Estado da Bahia / Frequency of antiphospholipid antibodies in patients with systemic lupus erythematosus in the State of Bahia

Objetivo: Estudar a prevalência dos anticorpos antifosfolípides e sua associação com complicações trombóticas em diferentes grupos raciais da Bahia. Métodos: Foram estudados 102 pacientes com diagnóstico de lúpus eritematoso sistêmico (LES) acompanhados ambulatoriamente. Dos 102 pacientes estudados, 17 eram da raça negra, 24 da raça branca e 61 eram mulatos. Cada paciente foi submetido a avaliação clínica e laboratorial, esta consistindo de exames gerais e especiais para detecção de anticorpos antifosfolípides: anticoagulante lúpico (testes de coagulação), anticardiolipina e anti-VDRL (ELISA). Resultados: Tromboses foram observadas em 5,9 por cento dos pacientes, plaquetopenia em 8,8 por cento e abortamento em 12,5 por cento. A presença de uma ou mais dessas manifestações foi observada em 20,6 por cento (21/102) dos pacientes, sendo vista em 23,5 por cento dos negros, 20 por cento dos brancos e 19,7 por cento dos mulatos (p=0,9). Anticorpos antifosfolípides foram detectados em 24,5 por cento dos casos (25/102). Analisando-se cada raça separadamente, observou-se uma frequência de 5,9 por cento de positividade na raça negra (1/17), 16,7 por cento na raça branca (4/24) e 32,8 por cento em mulatos (20/61), sendo essa diferença estatisticamente significante (p<0,05). Não se demonstrou uma associação entre a positividade desses anticorpos e a presença de complicações clínicas (p=0,5) quando todo o grupo foi analisado, porém houve associação entre a presença de tromboses e anticoagulante lúpico nos mulatos (p=0,02), mas o pequeno número de pacientes incluídos nessa análise exige cautela na interpretação desses resultados. Do mesmo modo, não se pode excluir a possibilidade de que a não associação entre morbidade gestacional e a presença desses anticorpos possa ser decorrente da limitação do desenho do estudo. Conclusões: a)A prevalência de anticorpos antifosfolípides pode variar com a raça. b) Analisando o grupo de portadores de LES como um todo, não se observou a associação da presença de anticorpos antifosfolípides com complicações trombóticas, abortamento e plaquetopenia