Cl gene cluster encoding several small nucleolar RNAs: a comparison amongst trypanosomatids

Small nucleolar RNAs (snoRNAs) are small non-coding RNAs that modify RNA molecules such as rRNA and snRNA by guiding 2'-O-ribose methylation (C/D box snoRNA family) and pseudouridylation reactions (H/ACA snoRNA family). H/ACA snoRNAs are also involved in trans-splicing in trypanosomatids. The aims of this work were to characterise the Cl gene cluster that encodes several snoRNAs in Trypanosoma rangeli and compare it with clusters from Trypanosoma cruzi, Trypanosoma brucei, Leishmania major, Leishmania infantum, Leishmania braziliensis and Leptomonas collosoma. The T. rangeli Cl gene cluster is an 801 base pair (bp) repeat sequence that encodes three C/D (Cl1, Cl2 and Cl4) and three H/ACA (Cl3, Cl5 and Cl6) snoRNAs. In contrast to T. brucei, the Cl3 and Cl5 homologues have not been annotated in the Leishmania or T. cruzi genome projects (http//:www.genedb.org). Of note, snoRNA transcribed regions have a high degree of sequence identity among all species and share gene synteny. Collectively, these findings suggest that the Cl cluster could constitute an interesting target for therapeutic (gene silencing) or diagnostic intervention strategies (PCR-derived tools).

Inhibition of Trypanosoma cruzi proline racemase affects host-parasite interactions and the outcome of in vitro infection

Proline racemase is an important enzyme of Trypanosoma cruzi and has been shown to be an effective mitogen for B cells, thus contributing to the parasite's immune evasion and persistence in the human host. Recombinant epimastigote parasites overexpressing TcPRAC genes coding for proline racemase present an augmented ability to differentiate into metacyclic infective forms and subsequently penetrate host-cells in vitro. Here we demonstrate that both anti T. cruzi proline racemase antibodies and the specific proline racemase inhibitor pyrrole-2-carboxylic acid significantly affect parasite infection of Vero cells in vitro. This inhibitor also hampers T. cruzi intracellular differentiation.

Proline racemases: insights into Trypanosoma cruzi peptides containing D-proline

Trypanosoma cruzi proline racemases (TcPRAC) are homodimeric enzymes that interconvert the L and D-enantiomers of proline. At least two paralogous copies of proline racemase (PR) genes are present per parasite haploid genome and they are differentially expressed during T. cruzi development. Non-infective epimastigote forms that overexpress PR genes differentiate more readily into metacyclic infective forms that are more invasive to host cells, indicating that PR participates in mechanisms of virulence acquisition. Using a combination of biochemical and enzymatic methods, we show here that, in addition to free D-amino acids, non-infective epimastigote and infective metacyclic parasite extracts possess peptides composed notably of D-proline. The relative contribution of TcPRAC to D-proline availability and its further assembly into peptides was estimated through the use of wild-type parasites and parasites over-expressing TcPRAC genes. Our data suggest that D-proline-bearing peptides, similarly to the mucopeptide layer of bacterial cell walls, may be of benefit to T. cruzi by providing resistance against host proteolytic mechanisms.

Biological and genetic aspects of crosses between species of the genus Meccus (Hemiptera: Reduviidae Triatominae)

The degree of reproductive isolation between Meccus phyllosomus and the remaining five species of the genus Meccus, as well as between Meccus bassolsae and Meccus pallidipennis, Meccus longipennis and Meccus picturatus, was examined. Fertility and the segregation of morphological characteristics were examined in two generations of hybrids from crosses between these species. The percentage of couples with offspring (fertile) was high in the vast majority of sets of crosses, with the exception of that between ♀M. phyllosomus and ♂Meccus mazzottii. In sets of crosses involving M. bassolsae specimens, no first-generation (F1) individuals were morphologically similar to M. bassolsae, but instead shared the morphology of the other parental species. A similar phenomenon was observed in most sets of crosses involving M. phyllosomus. These results indicated that different degrees of reproductive isolation exist among the species of Meccus involved in this study. The biological evidence obtained in this study does not support the proposal that M. bassolsae is a full species. It could indicate that, on the contrary, it should be considered a subspecies of a single polytypic species. The biological evidence does support the proposal that M. phyllosomus is a full species.

A human astrocytoma cell line is highly susceptible to infection with Trypanosoma cruzi

Astrocytes play a vital role in neuronal protection, homeostasis, vascular interchange and the local immune response. Some viruses and parasites can cross the blood-brain barrier and infect glia. Trypanosoma cruzi, the aetiological agent of Chagas disease, can seriously compromise the central nervous system, mainly in immune-suppressed individuals, but also during the acute phase of the infection. In this report, the infective capacity of T. cruzi in a human astrocyte tumour-derived cell line was studied. Astrocytes exposed to trypomastigotes (1:10 ratio) produced intracellular amastigotes and new trypomastigotes emerged by day 4 post-infection (p.i.). At day 6 p.i., 93% of the cells were infected. Using flow cytometry, changes were observed in both the expression of major histocompatibility complex class I and II molecules and the chemokine secretion pattern of astrocytes exposed to the parasite. Blocking the low-density lipoprotein receptor on astrocytes did not reduce parasite intracellular infection. Thus, T. cruzi can infect astrocytes and modulate the immune response during central nervous system infection.

A decade of malaria during pregnancy in Brazil: what has been done concerning prevention and management

In Brazil, malaria remains a disease of major epidemiological importance because of the high number of cases in the Amazonian Region. Plasmodium spp infections during pregnancy are a significant public health problem with substantial risks for the pregnant woman, the foetus and the newborn child. In Brazil, the control of malaria during pregnancy is primarily achieved by prompt and effective treatment of the acute episodes. Thus, to assure rapid diagnosis and treatment for pregnant women with malaria, one of the recommended strategy for low transmission areas by World Health Organization and as part of a strategy by the Ministry of Health, the National Malaria Control Program has focused on integrative measures with woman and reproductive health. Here, we discuss the approach for the prevention and management of malaria during pregnancy in Brazil over the last 10 years (2003-2012) using morbidity data from Malaria Health Information System. Improving the efficiency and quality of healthcare and education and the consolidation of prevention programmes will be challenges in the control of malaria during pregnancy in the next decade.

Molecular findings from influenza A(H1N1)pdm09 detected in patients from a Brazilian equatorial region during the pandemic period

After the World Health Organization officially declared the end of the first pandemic of the XXI century in August 2010, the influenza A(H1N1)pdm09 virus has been disseminated in the human population. In spite of its sustained circulation, very little on phylogenetic data or oseltamivir (OST) resistance is available for the virus in equatorial regions of South America. In order to shed more light on this topic, we analysed the haemagglutinin (HA) and neuraminidase (NA) genes of influenza A(H1N1)pdm09 positive samples collected during the pandemic period in the Pernambuco (PE), a northeastern Brazilian state. Complete HA sequences were compared and amino acid changes were related to clinical outcome. In addition, the H275Y substitution in NA, associated with OST resistance, was investigated by pyrosequencing. Samples from PE were grouped in phylogenetic clades 6 and 7, being clustered together with sequences from South and Southeast Brazil. The D222N/G HA gene mutation, associated with severity, was found in one deceased patient that was pregnant. Additionally, the HA mutation K308E, which appeared in Brazil in 2010 and was only detected worldwide the following year, was identified in samples from hospitalised cases. The resistance marker H275Y was not identified in samples tested. However, broader studies are needed to establish the real frequency of resistance in this Brazilian region.

Oseltamivir-resistant influenza A(H1N1)pdm2009 strains found in Brazil are endowed with permissive mutations, which compensate the loss of fitness imposed by antiviral resistance

The 2009 pandemic influenza A virus outbreak led to the systematic use of the neuraminidase (NA) inhibitor oseltamivir (OST). Consequently, OST-resistant strains, carrying the mutation H275Y, emerged in the years after the pandemics, with a prevalence of 1-2%. Currently, OST-resistant strains have been found in community settings, in untreated individuals. To spread in community settings, H275Y mutants must contain additional mutations, collectively called permissive mutations. We display the permissive mutations in NA of OST-resistant A(H1N1)pdm09 virus found in Brazilian community settings. The NAs from 2013 are phylogenetically distinct from those of 2012, indicating a tendency of positive selection of NAs with better fitness. Some previously predicted permissive mutations, such as V241I and N369K, found in different countries, were also detected in Brazil. Importantly, the change D344N, also predicted to compensate loss of fitness imposed by H275Y mutation, was found in Brazil, but not in other countries in 2013. Our results reinforce the notion that OST-resistant A(H1N1)pdm09 strains with compensatory mutations may arise in an independent fashion, with samples being identified in different states of Brazil and in different countries. Systematic circulation of these viral strains may jeopardise the use of the first line of anti-influenza drugs in the future.

Whole-genome sequences of influenza A(H3N2) viruses isolated from Brazilian patients with mild illness during the 2014 season

The influenza A(H3N2) virus has circulated worldwide for almost five decades and is the dominant subtype in most seasonal influenza epidemics, as occurred in the 2014 season in South America. In this study we evaluate five whole genome sequences of influenza A(H3N2) viruses detected in patients with mild illness collected from January-March 2014. To sequence the genomes, a new generation sequencing (NGS) protocol was performed using the Ion Torrent PGM platform. In addition to analysing the common genes, haemagglutinin, neuraminidase and matrix, our work also comprised internal genes. This was the first report of a whole genome analysis with Brazilian influenza A(H3N2) samples. Considerable amino acid variability was encountered in all gene segments, demonstrating the importance of studying the internal genes. NGS of whole genomes in this study will facilitate deeper virus characterisation, contributing to the improvement of influenza strain surveillance in Brazil.

Os fatores de riscos alimentares para câncer colorretal relacionado ao consumo de carnes

Uma das estratégias metodológicas para realizar a prática baseada em evidências é a revisão integrativa, que neste estudo teve como objetivo buscar e sintetizar as evidências disponíveis na literatura científica sobre os fatores de riscos alimentares para o câncer colorretal relacionado ao consumo de carnes. As bases de dados LILACS, MEDLINE, CINAHL e COCHRANE Library foram consultadas e os estudos pertinentes ao consumo de carnes somaram seis. As metanálises demonstraram que a ingestão de carne vermelha está relacionada com o aumento do risco para câncer colorretal em 28% a 35%, enquanto a carne processada está associada ao risco elevado de 20% a 49%. As evidências apontam a carne vermelha, a carne processada e o total de carne consumida como fatores de risco para o desenvolvimento de pólipos e câncer colorretal. Não foi identificado estudo que indicasse a ingestão de frango e peixe como fatores de risco.

Heart failure as a predictor of functional dependence in hospitalized elderly

AbstractOBJECTIVEIdentify whether Heart Failure (HF) is a predictor of functional dependence for Basic Activities of Daily Living (BADL) in hospitalized elderly.METHODSWe investigated medical records and assessed dependence to BADL (by the Katz Index) of 100 elderly admitted to a geriatric ward of a university hospital. In order to verify if HF is a predictor of functional dependence, linear regression analyzes were performed.RESULTSThe prevalence of HF was 21%; 95% of them were dependent for BADLs. Bathing was the most committed ADL. HF is a predictor of dependence in hospitalized elderlies, increasing the chance of functional decline by 5 times (95% CI, 0.94-94.48), the chance of functional deterioration by 3.5 times (95% CI, 1.28-11.66; p <0.02) and reducing 0.79 points in the Katz Index score (p <0.05).CONCLUSIONHF is a dependency predictor of ADL in hospitalized elderly, who tend to be more dependent, especially for bathing.

Notas sobre o tipo de Trigona limao Smith (Hymenoptera, Apidae, Lestrimelitta)

O nome limao tem sido usado para espécies distintas do gênero Lestrimelitta. O material-tipo de Smith depositado no The Natural History Museum (Londres) foi examinado. A operária de L. limao é descrita e o lectótipo para Trigona limao Smith, 1863 é designado. Esta espécie encontra-se restrita aos cerrados da porção central do Brasil, de São Paulo ao Maranhão. Distingue-se de outras espécies pelos seguintes caracteres diagnósticos: vértice com pouquíssimas cerdas eretas, muito finas e curtas, restritas à região interocelar; bordo anterior do mesoscuto com cerdas eretas castanhas nas laterais; laterais do mesepisterno com cerdas eretas esparsas e restritas à porção ventral e curvatura entre porções lateral e ventral, tornando-se mais longas próximo à base da coxa média; contorno da abertura do espiráculo propodeal, aproximadamente 2,5 x mais longo do que largo; flancos do propódeo coberto por pilosidade decumbente esbranquiçada, fina e ramificada; esporão mesotibial muito reduzido; tergo metassomal 1 com cerdas enegrecidas nas porções laterais.

Três espécies novas de Lestrimelitta Friese (Hymenoptera, Apidae) da Costa Rica, Panamá e Guiana Francesa

Três espécies novas de abelhas do gênero Lestrimelitta são descritas: L. danuncia sp. nov. (da Costa Rica e Panamá), L. mourei sp. nov. (da Costa Rica) e L. glaberrima sp. nov. (da Guiana Francesa). Todas essas espécies foram identificadas anteriormente como Lestrimelitta limao Smith.

Revisão taxonômica das espécies brasileiras de abelhas do gênero Lestrimelitta Friese (Hymenoptera, Apidae, Meliponina)

Revisão taxonômica das espécies de abelhas do gênero Lestrimelitta Friese (Hymenoptera, Apidae, Meliponina) que ocorrem no Brasil. Com base em caracteres morfológicos, como aqueles relacionados com a pilosidade, o formato do espiráculo propodeal, as distâncias interorbitais e o comprimento do esporão mesotibial, são reconhecidas catorze espécies, seis das quais novas para a Ciência: L. ciliata. sp. nov., L. maracaia sp. nov., L. similis sp. nov., L. spinosa sp. nov., L sulina. sp. nov. e L. tropica sp. nov. São designados lectótipos para Trigona (Lestrimelitta) limao var. rufipes Friese, 1903 e Trigona (Lestrimelitta) limao var. rufa Friese, 1903. O macho de L. limao (Smith, 1863) é descrito pela primeira vez. É apresentada também uma diagnose das seguintes espécies: L. ehrhardti Friese, 1931; L. glaberrima Oliveira & Marchi, 2005; L. glabrata Camargo & Moure, 1989; L. monodonta Camargo & Moure, 1989; e L. nana Melo, 2003. São apresentados chaves de identificação para operárias e machos, ilustrações e mapas de ocorrência.

Dinâmica de nitrogênio em lodo de esgoto sob condições de estocagem

Este estudo teve como objetivo avaliar os teores de N-total, N-NH4+, N-NO3- e pH em colunas de lodo de esgoto anaeróbio alcalinizado e na solução percolada, simulando condições reais de armazenamento. O experimento foi desenvolvido em campo, no município de Curitiba (PR), em clima Cfb e pluviosidade de 197 mm durante o período de incubação, que correspondeu a 48 dias. Os fatores testados foram: três tipos de lodo (lodo alcalinizado coberto - LCC; lodo alcalinizado descoberto - LCD, e lodo bruto descoberto - LBD), quatro intervalos de incubação (0; 15; 27; e 48 dias) e cinco profundidades na coluna (0-5; 5-20; 20-50; 50-80; e composta 0-80 cm), com quatro repetições. O líquido percolado foi avaliado no 1º, 5º, 9º, 13º, 27º, 36º e 48º dia. A alcalinização (CaO) foi realizada à proporção de 50 % da matéria seca (MS) do lodo. A cobertura (vedação com lona plástica e tampo de amianto em LCC) teve a finalidade de conter a volatilização de gases, especialmente amônia (NH3). As condições do meio ocasionadas pela alcalinização do lodo reduziram significativamente a intensidade de mineralização de N e a nitrificação. Os lodos brutos e alcalinizados em condição de estocagem apresentaram perdas consideráveis de N-NH4+ e N-org. Os efeitos diluição, volatilização de NH3 e lixiviação organomineral conseqüentes da alcalinização (50 % MS lodo) reduziram os teores de N-total em até 50 %. O processo de alcalinização potencializou perdas de N por meio da volatilização de NH3 e lixiviação de N-NH4+. Entretanto, a cobertura reduziu as perdas de N sob a forma de NH3.