Next-Generation Sequencing for Molecular Diagnosis of Cystic Fibrosis in a Brazilian Cohort.

Cystic fibrosis (CF), an autosomal recessive genetic disease, is recognized as one of the most prevalent diseases in Caucasian populations. Epidemiological data show that the incidence of CF varies between countries and ethnic groups in the same region. CF occurs due to pathogenic variants in the gene encoding cystic fibrosis transmembrane conductance regulator (CFTR), located on chromosome 7q31.2. To date, more than 2,000 variants have been registered in the CFTR database. The study of these variants leads to the diagnosis and the possibility of a specific treatment for each patient through precision medicine. In this study, complete screening of CFTR was performed through next-generation sequencing (NGS) to gain insight into the variants circulating in the population of Rio de Janeiro and to provide patient access to treatment through genotype-specific therapies. Samples from 93 patients with an inconclusive molecular diagnosis were subjected to full-length screening of CFTR using an Illumina NGS HiSeq platform. Among these patients, 46 had two pathogenic variants, whereas 12 had only one CFTR variant. Twenty-four variants were not part of our routine screening. Of these 24 variants, V938Gfs∗37 had not been described in the CF databases previously. This research achieved a molecular diagnosis of the patients with CF and identification of possible molecular candidates for genotype-specific treatments.

Draft Genome Sequences of Leptospira interrogans Serovar Copenhageni Strains Isolated from Patients with Weil's Disease in Brazil.

Leptospirosis is a worldwide zoonosis caused by pathogenic species of Leptospira In Brazil, this disease is endemic, presenting epidemic potential in rainy seasons. Here, we announce the whole-genome sequences of two L. interrogans serovar Copenhageni strains isolated from blood samples from two icteric patients associated with severe leptospirosis in Brazil.

Genetic information on OXA-23-producing Acinetobacter baumannii strain CCBH15815, belonging to ST730/ST783, isolated from Brazil.

In Brazil, A. baumannii has been described as nosocomial pathogens causing hospital-acquired infections. Current WGS technologies have been useful in identifying of genetic features between Acinetobacter isolates. Here, we report the draft genome sequence of OXA-23 producing A. baumannii CCBH15815 clinical isolate, belonging to ST730/ST783, recovered from a 21-year-old hospitalised patient. We observed important resistance determinant genes, especially beta-lactamases-encoding genes, in an estimated genome size of 4,058,633 bp with 3839 predicted coding regions.

Bioactivity under laboratory conditions of Brevibacillus laterosporus towards larvae and adults of Chrysomya putoria (Diptera: Calliphoridae).

Brevibacillus laterosporus was tested for entomopathogenic activity towards larvae and adults of Chrysomya putoria (Diptera: Calliphoridae) under laboratory conditions. Sublethal effects related to feeding activity or development were observed, including reduction in larval weight gain, probably by inhibition of feeding, and variation in the duration of the developmental stages of the insect. Larval mortality was dose dependent following ingestion. The experiments were performed with newly emerged adults exposed to a sugar based diet containing spore suspensions. Concentrations of 1.13 × 109 CFU/ml caused 70.5% of mortality. The present study highlights the potential of B. laterosporus to control populations of C. putoria, a dipteran of medical-veterinary and sanitary importance, both in larval and adult stages.

Whole genome sequence of Mycobacterium kansasii isolates of the genotype 1 from Brazilian patients with pulmonary disease demonstrates considerable heterogeneity.

Mycobacterium kansasii is an opportunistic pathogen and one of the most commonly encountered species in individuals with lung disease. We here report the complete genome sequence of 12 clinical isolates of M. kansasii from patients with pulmonary disease in Brazil.

Whole genome sequence of Mycobacterium kansasii isolates of the genotype 1 from Brazilian patients with pulmonary disease demonstrates considerable heterogeneity

Mycobacterium kansasii is an opportunistic pathogen and one of the most commonly encountered species in individuals with lung disease. We here report the complete genome sequence of 12 clinical isolates of M. kansasii from patients with pulmonary disease in Brazil.

Next-generation sequencing virulome analysis of a Yersinia enterocolitica subsp. palearctica bioserotype 4/O: 3 ST18 isolated from human blood in Brazil

Abstract Yersinia enterocolitica is a widespread Gram-negative bacterium that causes gastrointestinal disease and other clinical manifestations in humans. Potentially pathogenic Y. enterocolitica has been isolated in Brazil, from human, environmental, food, and animal sources. Herein we report a genome sequence of Y. enterocolitica subsp. palearctica strain YE 19, serotype O:3, biotype 4, sequence type 18, with virulence determinants isolated from human blood in Rio de Janeiro in 2005. The results corroborate other findings that this strain harbors a set of virulence determinants that could play a role in host pathoadaptation and may also justify the successful dissemination of bioserotype 4/O:3 in Brazil. The presence of strains harboring all of these virulence genes in Brazil is a potential threat to young children and immunocompromised individuals, for whom yersiniosis are a significant source of morbidity and mortality. The results of a genomic data analysis will help understand the virulence of Brazilian strains and provide data for Y. enterocolitica studies worldwide.

Next-generation sequencing virulome analysis of a Yersinia enterocolitica subsp. palearctica bioserotype 4/O:3 ST18 isolated from human blood in Brazil.

Yersinia enterocolitica is a widespread Gram-negative bacterium that causes gastrointestinal disease and other clinical manifestations in humans. Potentially pathogenic Y. enterocolitica has been isolated in Brazil, from human, environmental, food, and animal sources. Herein we report a genome sequence of Y. enterocolitica subsp. palearctica strain YE 19, serotype O:3, biotype 4, sequence type 18, with virulence determinants isolated from human blood in Rio de Janeiro in 2005. The results corroborate other findings that this strain harbors a set of virulence determinants that could play a role in host pathoadaptation and may also justify the successful dissemination of bioserotype 4/O:3 in Brazil. The presence of strains harboring all of these virulence genes in Brazil is a potential threat to young children and immunocompromised individuals, for whom yersiniosis are a significant source of morbidity and mortality. The results of a genomic data analysis will help understand the virulence of Brazilian strains and provide data for Y. enterocolitica studies worldwide.

Functional diversity of the plant glycine-rich proteins superfamily.

The first plant glycine-rich proteins (GRPs) have been isolated more than 20 years ago based on their specific expression pattern and/or modulation by several biotic and abiotic factors. This superfamily is characterized by the presence of a glycine-rich domain arranged in (Gly)(n)-X repeats. The presence of additional motifs, as well as the nature of the glycine repeats, groups them in different classes. The diversity in structure as well as in expression pattern, modulation and sub-cellular localization have always indicated that these proteins, although classified as members of the same superfamily, would perform different functions in planta. Only now, two decades later, with the first functional characterizations of plant GRPs their involvement in diverse biological and biochemical processes are being uncovered. Here, we review the so far ascribed functions of plant GRPs.

Association between BanI genotype and increased phospholipase A2 activity in schizophrenia.

Phospholipases A2 (PLA2) are a family of key enzymes in the metabolism of membrane phospholipids. Several studies reported on increased blood and brain PLA2 activity in schizophrenia, which suggest a disordered phospholipid metabolism in the disease. In addition, a genetic variant of a cytosolic PLA2 gene has been reported to be associated with schizophrenia. These data indicate that variants of PLA2 encoding genes are plausible candidates for increasing the susceptibility for schizophrenia. In this study, we investigated a possible association between PLA2 activity in platelets and a polymorphic site for BanI in the PLA2 (group 4A) gene on chromosome 1q25. Seventy-five schizophrenic patients (DSM-IV) and 68 healthy controls were recruited and the PCR assays were performed. A radioenzymatic assay for the cytosolic PLA2 activity in platelets was used. The allele A2 and the genotype A2A2 were more frequent in schizophrenic patients than in controls (p<0.005 and p<0.05 respectively). When we assorted the subjects according to their genotypes, we found that PLA2 activity was significantly higher in patients with the A2A2 genotype (29.6+/-5.1 pMol/mg protein/min) than in those with the A1A2 (20.8+/-3.6 pMol/mg protein/min, p<0.001) or A1A1 genotype (15.9+/-5.1 pMol/mg protein/min, p<0.001). Also in controls, carriers of the A2 allele (A1A2 and A2A2) had higher PLA2 activity than the A1A1 group (p=0.004 for both). Our data suggest an association between BanI genotype and PLA2G4A activity in platelets and that the presence of the allele A2 may increase risk for schizophrenia through an increment of PLA2 activity.

Survey of glycine-rich proteins (GRPs) in the Eucalyptus expressed sequence tag database (ForEST)

The occurrence of quasi-repetitive glycine-rich peptides has been reported in different organisms. Glycine-rich regions are proposed to be involved in protein-protein interactions in some mammalian protein families. In plants, a set of glycine-rich proteins (GRPs) was characterized several years ago, and since then a wealth of new GRPs have been identified. GRPs may have very diverse sub-cellular localization and functions. The only common feature among all different GRPs is the presence of glycine-rich repeat domains. The expression of genes encoding GRPs is developmentally regulated, and also induced, in several plant genera, by physical, chemical and biological factors. In addition to the highly modulated expression, several GRPs also show tissue-specific localization. GRPs specifically expressed in xylem, phloem, epidermis, anther tapetum and roots have been described. In this paper, the structural and functional features of these proteins in Eucalyptus are summarized. Since this is the first description of GRPs in this species, particular emphasis has been given to the expression pattern of these genes by analyzing their abundance and prevalence in the different cDNA-libraries of the Eucalyptus Genome Sequencing Project Consortium (ForEST). The comparison of GRPs from Eucalyptus and other species is also discussed

Ocorrência de enteroparasitoses em alunos da escola pública de ensino fundamental do município de Catanduva (São Paulo, Brasil) / Survey of parasitic diseases in students at a public primary school at Catanduva city (São Paulo, Brazil)

Este trabalho objetivou levantar as parasitoses intestinais mais freqüentes na faixa etária de 6 a 11 anos, relacionando-as ao nível sócio-econômico e aos hábitos de vida das crianças de uma escola pública de primeiro grau no município de Catanduva. Iniciou-se com palestras sobre educação sanitária para pais e alunos. Posteriormente, foi aplicado um questionário sócio-econômico e colheram-se amostras de fezes de 250 crianças, sendo 138 do sexo masculino e 112 do sexo feminino. Foram feitos exames parasitológicos de fezes; as crianças, que apresentaram resultados positivos, foram tratadas. Das crianças analisadas, 12,80% estavam parasitadas, principalmente aquelas na faixa etária de 8 a 9 anos (40,62%) e 62,50% eram do sexo masculino. As 84,37% de crianças parasitadas das crianças parasitadas e 67,88% das crianças não parasitadas relataram que tinham animais de estimação. Os parasitas mais encontrados foram Giardia lamblia (43,75%); Entamoeba coli (31,25%); Enterobius vermicularis (18,75%); Endolimax nana (15,62%); ancilostomídeos (3,12%); Strongyloides stercoralis (3,12%) e Entamoeba hystolitica (3,12%).Das crianças positivas, 81,25% eram monoparasitadas e 18,7% poliparasitadas ; e 77,27% bebiam água diretamente da torneira. Concluiu-se que a incidência de parasitoses diminuiu no município de Catanduva se comparada a dos anos anteriores, mostrando ter havido melhoria na qualidade de vida e maior conhecimento das pessoas em relação à educação sanitária e saneamento básico

Allelic association analysis of phospholipase A2 genes with schizophrenia.

Several studies suggest increased activity of phospholipase A2 in schizophrenic patients. In the present study, variants of four genes coding for phospholipase A2 enzyme groups (sPLA2, cPLA2, iPLA2 and PAFAH) were analysed in a case-control sample using 240 schizophrenic patients and 312 healthy controls. No difference was observed on the allelic and genotypic distribution of cPLA2 and sPLA2 gene polymorphisms among the groups. The PAFAH variant was very rare in our population and therefore not informative. A significant allelic (chi2=5.86, P=0.0085, odds ratio=1.38, 95% confidence interval, 1.08-1.77) and genotypic (chi2=5.4, P=0.02) association with the iPLA2 gene polymorphism was found. In conclusion, our data suggested that iPLA2 may play a role as a susceptibility gene for schizophrenia in our sample; however, confirmatory studies in independent samples are needed.

Allelic association study between phospholipase A2 genes and bipolar affective disorder.

OBJECTIVES: In vivo studies demonstrating that lithium is a powerful phospholipase A2 (PLA2) inhibitor suggest that PLA2 activation, and subsequent cell signaling overactivation by increased fatty acid release may be the primary abnormality in bipolar affective disorder (BPAD), thus making PLA2 genes attractive candidates for the susceptibility to BPAD. The present study investigates polymorphisms in cytosolic phospholipase A2 (cPLA2), calcium-independent phospholipase A2 (iPLA2), and secretory phospholipase (sPLA2) genes in a Brazilian sample. METHODS: A cross-sectional study was performed with 181 unrelated DSM-IIIR BPAD subjects and 312 controls. A polymerase chain reaction-restriction fragment length polymorphism assay for BanI cPLA2 and AvrII iPLA2 polymorphisms was performed, and an ATT repeat in sPLA2 was assessed using a semiautomated genetic analyzer (ALFexpress). RESULTS: There was no significant difference observed in the allelic and genotypic distribution between the BPAD and control groups for cPLA2 (genotype: chi2 = 0.8, 2df, p = 0.6; allele chi2 = 0, 1df, p = 0.9), iPLA2 (genotype: chi2 = 1.7, 2df, p = 0.4; allele: chi2 = 0.3, 1df, p = 0.6), and sPLA2 (allele: chi2 = 3.6, 6df, p = 0.8). CONCLUSION: Our results failed to demonstrate that the studied PLA2 polymorphisms were associated with an increased risk for BPAD in our sample.

Estudo de associaçäo entre o polimorfismo serina-9-glicina do receptor dopaminérgico D3 e esquizofrenia / Study of association between the ser-9-gly polymorphism of the D3 dopaminergic receptor and schizophrenia

Duas abordagens genético-moleculares foram realizadas para investigar a possível associaçäo entre o polimorfismo serina-9-glicina no receptor dopaminérgico D3 e esquizofrenia. Na primeira análise, um grupo de 141 pacientes com esquizofrenia foi comparado a um grupo-controle de 189 indivíduos pareado para sexo e origem étnica. No outro estudo, foi realizada análise de 35 trios (pai e mäe näo afetados e paciente com esquizofrenia). Os resultados desses estudos näo apresentaram associaçäo alélica ou genotípica estatisticamente significante com esquizofrenia. Pode-se concluir que o polimorfismo serina-9-glicina no receptor dopaminérgico D3 näo é um fator de risco para desenvolver esquizofrenia na populaçäo estudada

Sugarcane expressed sequences tags (ESTs) encoding enzymes involved in lignin biosynthesis pathways

Ligninas säo compostos fenólicos encontrados nas paredes secundárias do sistema vascular vegetal e desempenham importantes papéis biológicos, reduzindo a permeabilidade da parede celular em relaçäo à água, estando também envolvidas em mecanismos de defesa contra patógenos. A via metabólica da lignina e as enzimas envolvidas na sua síntese vem sendo caracterizadas nos últimos anos. Uma série de genes que codificam diferentes enzimas envolvidas na biossíntese da lignina foram identificados em diferentes espécies de plantas. A biossíntese de lignina está acoplada ao metabolismo dos fenilpropanóides, apresentando enzimas compartilhadas com outros processos metabólicos tais como fenilalanina amônio liase (PAL), cinnamato 4-hidroxilase (C4H) and ácido caféico O-metiltransferase (COMT) assim como enzimas específicas como cinamoil-CoA redutase (CCR) e cinamil álcool dehidrogenase (CAD). Em certos tipos de mutantes de milho e sorgo, um aumento na digestibilidade foi associado a uma reduçäo no teor de lignina. Uma reduçäo na atividade de CAD e COMT, importantes enzimas envolvidas na biossíntese de lignina vem sendo demonstrada. Estas observações tem motivado diferentes grupos de pesquisa e alterarem o conteúdo ou a composiçäo da lignina em plantas modelo, assim como culturas de interesse econômico, através de engenharia genética. O principal objetivo prático destes projetos é uma otimizaçäo da utilizaçäo destas plantas levando a uma maior produçäo de papel e digestibilidade da raçäo animal. No presente trabalho foi realizado um inventário das seqüências de ESTs, que codificam enzimas envolvidas no metabolismo de lignina, presentes no Projeto Genoma de Cana-de-açúcar (SUCEST). A análise foi realizada com as enzimas chaves ferulato-5-hidroxilase (F5H), ácido caféico O-metiltransferase (COMT), cafeoil CoA O-metiltransferase (CCoAOMT), hidroxicinamato CoA ligase (4CL), cinamoil-CoA redutase (CCR) and cinamil álcool dehidrogenase (CAD). A análise comparativa destes genes com os descritos em outras espécies poderá servir para a identificaçäo de marcadores moleculares em programas de melhoramento genético, assim como em projetos que visem a manipulaçäo do metabolismo de lignina em cana-de-açúcar.

Classification, expression pattern and comparative analysis of sugarcane expressed sequences tags (ESTs) encoding glycine-rich proteins (GRPs)

Desde o isolamento da primeira proteína rica em glicina (GRP) em plantas, um grande número de novas GRPs vem sendo identificado. Seu padräo de expressäo altamente específico, embora variado, em conjunto com as diferentes localizações sub-celulares de alguns dos tipos de GRPs, claramente indica que estas proteínas encontram-se envolvidas em diversos processos fisiológicos independentes. Embora ainda sem uma clara definiçäo do papel de GRPs na célula vegetal, estudos realizados com estas proteínas têm resultado em novos e interessantes esclarecimentos da biologia celular e molecular de plantas. Promotores com regulaçäo complexa, assim como distintos mecanismos de regulaçäo da expressäo gênica tem sido demonstrados. Novas vias de endereçamento de proteínas tais como a exportaçäo de GRPs para diferentes tipos celulares, tem sido observados. Estes dados mostram que as GRPs podem constituir marcadores e/ou modelos interessantes para a compreensäo de distintos aspectos da biologia vegetal. Neste trabalho, características estruturais e funcionais deste tipo de proteínas em cana-de-açúcar (Saccharum officinarum L.) foram analisadas. Uma vez que esta é a primeira descriçäo deste tipo de proteínas, em cana-de-açúcar, especial atençäo foi dada para o padräo de expressäo destes genes, analisando-se a abundância e prevalência de cada um dos genes nas diferentes bibliotecas de cDNA do projeto Sequenciamento de ESTs de cana-de-açúcar (SUCEST). A comparaçäo das GRPs de cana-de-açúcar com as GRPs descritas em outras espécies também será discutida.