Gastroenterites e infecções respiratórias agudas em crianças menores de 5 anos, em área da região Sudeste do Brasil, 1986-1987: II - Diarréias

INTRODUCTION: The decrease morbi-mortality gastroenteritis in is related to the factor responsible largely for the fall in infant mortality and mortality from communicable diseases in developing countries. Nevertheless, diarrhea is still a considerable public health problem in these countries, especially among children under 5 years old. OBJECTIVES: To describe some aspects of the of gastroenteritis epidemiology among children up to 5 years old, resident in areas of S. Paulo county. MATERIAL AND METHOD: A probabilistic sample of children up to 5 years old was studied (n=468). The epidemiological survey was undertaken in five areas S. Paulo county (Brazil) from March 1986 to May 1987. Data were obtained through household interviews once a month over a year. RESULTS: During the follow-up 139 diarrhea episodes were registered, with a mean duration of 5.5 days. Twenty percent of the diarrhea events were followed by at least one other case in the household. The incidence of gastroenteritis was 2.78 episodes per 100 children/month. The highest incidence affected the children of up to 2 years of age. In 46.1% of the gastroenteritis episodes medical assistance was not sought, the children were treated by their mothers or not at all; 51.8% of the diarrhea events were attended to by the primary health care service, and only 2.1% were attended to by a hospital. No child died as a consequence of diarrhea. Of the therapeutical interventions used the most frequent were oral rehydration (25.2%) and oral rehydration with antibiotics (11.5%). Various socio-economic and personal background factors such as living conditions, water supply, sewarage, coverage; family income per capita and prior history of frequent gastroenteritis were associated with a higher incidence of gastroenteritis. DISCUSSION: The results seen to reflect the tendency of morbi-mortality by gastroenteritis to decrease in S. Paulo county during the 1980's when hospital treatment of diarrhea presented a considerable reduction. This tendency must be observed closely, because it will influence the changes to be mode in the kind of demand for medical care.INTRODUÇÃO: O declínio da morbi-mortalidade pelas gastroenterites é, em boa parte, responsável pela queda da mortalidade infantil e da mortalidade por doenças infecciosas nos países do terceiro mundo. Esse agravo ainda se destaca, nesses países, como importante problema de saúde pública, especialmente, entre os menores de 5 anos. OBJETIVOS: Descrever aspectos do comportamento das gastroenterites entre crianças menores de 5 anos, residentes em 5 bairros do Município de São Paulo. MATERIAL E MÉTODO: Estudou-se uma amostra probabilística (N = 468) de crianças menores de 5 anos, residentes em 5 áreas do Município de São Paulo, SP (Brasil), acompanhada durante um ano, por meio de entrevistas mensais. RESULTADOS: Durante o acompanhamento foram identificados 139 episódios de diarréia, com uma duração média de 5,5 dias, 10% dos casos prolongaram-se por 15 dias ou mais. Em 20% dos episódios havia ao menos outra pessoa na família com diarréia. A incidência foi de 2,78 casos por 100 crianças/mês, sendo mais elevada nos menores de 2 anos. Em 46,1% dos episódios de gastroenterite as crianças não demandaram assistência médica tendo sido tratadas pelas próprias mães, ou não receberam qualquer tratamento; em 51,8% dos episódios o atendimento foi feito em serviços de assistência primária à saúde e somente 2,1% dos casos necessitaram tratamento hospitalar. Nenhuma criança evoluiu para óbito. Entre as medidas terapêuticas mais utilizadas estão a reidratação oral (25,2%) e a antibioticoterapia associada à reidratação oral (11,5%); em somente 2 casos foi feita reidratação endovenosa. Alguns fatores socioeconômicos e antecedentes pessoais mostraram-se associados à ocorrência de diarréias, entre eles, as condições da habitação, saneamento básico e renda familiar "per capita" e história pregressa de diarréias freqüentes. DISCUSSÃO: Os resultados obtidos parecem refletir a tendência de diminuição da morbi-mortalidade por diarréias no Município de São Paulo, durante a década de 80, período em que houve acentuada queda nas internações hospitalares por essa causa. Tal tendência deve ser acompanhada atentamente, pois influenciará modificações nas características da demanda de assistência à saúde infantil

Glucocorticoid: Major Factor for Reduced Immunogenicity of 2009 Influenza A (H1N1) Vaccine in Patients with Juvenile Autoimmune Rheumatic Disease

Objective. To assess the immunogenicity and safety of non-adjuvanted influenza A H1N1/2009 vaccine in patients with juvenile autoimmune rheumatic disease (ARD) and healthy controls, because data are limited to the adult rheumatologic population. Method's. A total of 237 patients with juvenile ARD [juvenile systemic lupus erythematosus (JSLE), juvenile idiopathic arthritis (JIA), juvenile dermatomyositis (JDM), juvenile scleroderma, and vasculitis] and 91 healthy controls were vaccinated. Serology for anti-H1N1 was performed by hemagglutination inhibition assay. Seroprotection rate, seroconversion rate, and factor-increase in geometric mean titer (GMT) were calculated. Adverse events were evaluated. Results. Age was comparable in patients and controls (14.8 +/- 3.0 vs 14.6 +/- 3.7 years, respectively; p = 0.47). Three weeks after immunization, seroprotection rate (81.4% vs 95.6%; p = 0.0007), seroconversion rate (74.3 vs 95.6%; p < 0.0001), and the factor-increase in GMT (12.9 vs 20.3; p = 0.012) were significantly lower in patients with juvenile ARD versus controls. Subgroup analysis revealed reduced seroconversion rates in JSLE (p < 0.0001), JIA (p = 0.008), JDM (p = 0.025), and vasculitis (p = 0.017). Seroprotection (p < 0.0001) and GMT (p < 0.0001) were decreased only in JSLE. Glucocorticoid use and lymphopenia were associated with lower seroconversion rates (60.4 vs 82.9%; p = 0.0001; and 55.6 vs 77.2%; p = 0.012). Multivariate logistic regression including diseases, lymphopenia, glucocorticoid, and immunosuppressants demonstrated that only glucocorticoid use (p = 0.012) remained significant. Conclusion. This is the largest study to demonstrate a reduced but adequate immune response to H1N1 vaccine in patients with juvenile ARD. It identified current glucocorticoid use as the major factor for decreased antibody production. The short-term safety results support its routine recommendation for patients with juvenile ARD. ClinicalTrials.gov; NCT01151644. (First Release Nov 15 2011; J Rheumatol 2012;39:167-73; doi:10.3899/jrheum.110721)Fundacao de Amparo a Pesquisa do Estado de Sao Paulo (FAPESP) [2010/10749-0]Conselho Nacional de Desenvolvimento Cientifico e Tecnologico (CNPQ) [300248/2008-3, 300665/2009-1, 301411/2009-3, 300559/2009-7]Federico FoundationButantan Foundatio

Dengue Virus Type 4 Phylogenetics in Brazil 2011: Looking beyond the Veil

Dengue Fever and Dengue Hemorrhagic Fever are diseases affecting approximately 100 million people/year and are a major concern in developing countries. In the present study, the phylogenetic relationship of six strains of the first autochthonous cases of DENV-4 infection occurred in Sao Paulo State, Parana State and Rio Grande do Sul State, Brazil, 2011 were studied. Nucleotide sequences of the envelope gene were determined and compared with sequences representative of the genotypes I, II, III and Sylvatic for DEN4 retrieved from GenBank. We employed a Bayesian phylogenetic approach to reconstruct the phylogenetic relationships of Brazilian DENV-4 and we estimated evolutionary rates and dates of divergence for DENV-4 found in Brazil in 2011. All samples sequenced in this study were located in Genotype II. The studied strains are monophyletic and our data suggest that they have been evolving separately for at least 4 to 6 years. Our data suggest that the virus might have been present in the region for some time, without being noticed by Health Surveillance Services due to a low level of circulation and a higher prevalence of DENV-1 and DENV- 2

Genomic Surveillance of Yellow Fever Virus Epizootic in São Paulo, Brazil, 2016 – 2018

São Paulo, a densely inhabited state in southeast Brazil that contains the fourth most populated city in the world, recently experienced its largest yellow fever virus (YFV) outbreak in decades. YFV does not normally circulate extensively in São Paulo, so most people were unvaccinated when the outbreak began. Surveillance in non-human primates (NHPs) is important for determining the magnitude and geographic extent of an epizootic, thereby helping to evaluate the risk of YFV spillover to humans. Data from infected NHPs can give more accurate insights into YFV spread than when using data from human cases alone. To contextualise human cases, identify epizootic foci and uncover the rate and direction of YFV spread in São Paulo, we generated and analysed virus genomic data and epizootic case data from NHPs in São Paulo. We report the occurrence of three spatiotemporally distinct phases of the outbreak in São Paulo prior to February 2018. We generated 51 new virus genomes from YFV positive cases identified in 23 different municipalities in São Paulo, mostly sampled from NHPs between October 2016 and January 2018. Although we observe substantial heterogeneity in lineage dispersal velocities between phylogenetic branches, continuous phylogeographic analyses of generated YFV genomes suggest that YFV lineages spread in São Paulo at a mean rate of approximately 1km per day during all phases of the outbreak. Viral lineages from the first epizootic phase in northern São Paulo subsequently dispersed towards the south of the state to cause the second and third epizootic phases there. This alters our understanding of how YFV was introduced into the densely populated south of São Paulo state. Our results shed light on the sylvatic transmission of YFV in highly fragmented forested regions in São Paulo state and highlight the importance of continued surveillance of zoonotic pathogens in sentinel species

Caracterização de genótipos de rotavírus em creches: era pré- e pós-vacinação contra o rotavírus Characterization of rotavirus strains from day care centers: pre- and post-rotavirus vaccine era

OBJETIVOS: Em 2006, a vacina contra rotavírus foi incluída no Programa Nacional de Imunização. Este estudo teve como objetivo analisar os resultados da vigilância de genótipos de rotavírus em crianças < 5 anos com gastrenterites agudas provenientes de creches no estado de São Paulo por um período de 5 anos. MÉTODOS: Este estudo retrospectivo foi realizado em 30 creches no período de 2004 a 2008, com amostras fecais convenientes da vigilância das diarreias agudas, analisadas por ELISA, SDS-PAGE, RT-PCR e sequenciamento genético para caracterização do genótipo. RESULTADOS: Infecções por rotavírus foram detectadas em 28,3% de amostras (38/134). Os genótipos mais frequentes detectados foram: G9P[8] e G1P[8] em 2004; G1P[8] em 2005; GNTP[NT] em 2006; G2P[4] em 2007; e nenhum caso foi relatado em 2008. Infecções mistas não foram observadas. A taxa de detecção diminuiu de 65,7% (23/35) em 2004 para 50% (9/18) em 2007. CONCLUSÕES: A distribuição do genótipo variou de acordo com os anos, acompanhada pela redução no número de casos detectados. É necessário intensificar a vigilância pós-implantação da vacina contra rotavírus, visando monitorar as linhagens circulantes e sua eficácia contra possíveis genótipos emergentes.<br>OBJECTIVES: In 2006 the rotavirus vaccine was included in the Brazilian Immunization Program. The aim of this study was to report the results of a 5-year surveillance study of rotavirus strains in children < 5 years with acute gastroenteritis from day care centers in the state of São Paulo, Brazil. METHODS: This retrospective study was conducted with 30 day care centers from 2004 to 2008 with convenient surveillance fecal specimens, investigated by ELISA, SDS-PAGE, RT-PCR and gene sequencing to genotype characterization. RESULTS: Rotavirus infection was detected in 28.3% of samples (38/134). The most frequent genotypes detected were G9P[8] and G1P[8] in 2004; G1P[8] in 2005; GNTP[NT] in 2006; G2P[4] in 2007; and there were no cases in 2008. Mixed infections were not observed. Detection rate declined from 65.7% (23/35) in 2004 to 50% (9/18) in 2007. CONCLUSIONS: Genotype distribution varied according to collection year, accompanied by a reduction in detection rate. Use of rotavirus vaccine requires implementation of post-marketing surveillance to monitor rotavirus strain diversity and its efficacy against possible new emerging genotypes

Caracterização molecular de cepas de rotavírus e norovírus: um estudo de 6 anos (2004-2009)

OBJETIVO: Monitorar infecções causadas por rotavírus (RV) e norovírus (NoV) em crianças hospitalizadas < 5 anos com gastroenterite aguda provenientes do estado de São Paulo durante um período de 6 anos (2004-2009). MÉTODOS: Este estudo retrospectivo foi realizado em 61 centros médicos, utilizando amostras fecais coletadas por conveniência, analisadas por ensaio imunoenzimático, eletroforese em gel de poliacrilamida, transcrição reversa seguida de reação em cadeia pela polimerase e sequenciamento para caracterização dos genótipos. RESULTADOS: Infecções por RV e NoV foram detectadas em 29,6% (144/487) e 29,2% (26/89) das amostras, respectivamente. Os genótipos de RV detectados com maior frequência foram: G9P[8] em 2004; G1P[8] em 2005; G9P[8] em 2006; e G2P[4] durante os anos de 2007, 2008 e 2009. A taxa de detecção diminuiu de 36,3% (33/91) em 2004 para 4,2% (4/95) em 2009. NoV pertencente ao genogrupo GII foi encontrado em 61,6% (16/26) das amostras, e GI em 11,5% (3/26). Infecções mistas por NoV e RV foram observadas em 2,2% (2/89) das amostras, envolvendo as cepas GI+G9P[8] e GI+G2P[4]. CONCLUSÕES: A distribuição dos genótipos de RV variou com os anos, acompanhada pela redução no número de casos detectados. Ė necessário intensificar a vigilância pós-implantação da vacina contra RV, visando monitorar as cepas circulantes e sua eficácia contra possíveis genótipos emergentes. Os NoVs têm sido cada vez mais identificados como agentes etiológicos relevantes entre crianças hospitalizadas e exercem um papel importante na etiologia viral da gastroenterite pediátrica aguda no estado de São Paulo

Development of a Microtiter Plate Hybridization-Based PCR-Enzyme-Linked Immunosorbent Assay for Identification of Clinically Relevant Human Group A Rotavirus G and P Genotypes▿

A microtiter plate hybridization-based PCR-enzyme-linked immunosorbent assay (PCR-ELISA) has been used for the detection and identification of a variety of microorganisms. Here, we report the development of a PCR-ELISA for the identification of clinically relevant human rotavirus VP7 (G1 to G6, G8 to G10, and G12) and VP4 (P[4], P[6], P[8], P[9], and P[14]) genotypes. The G and P types of reference human and animal rotavirus strains for which specific probes were available were correctly identified by the PCR-ELISA. In addition, reference strains bearing G or P genotypes for which specific probes were unavailable, such as G11, G14, P[3], P[10], and P[11], did not display any cross-reactivity to the probes. The usefulness of the assay was further evaluated by analyzing a total of 396 rotavirus-positive stool samples collected in four countries: Brazil, Ghana, Japan, and the United States. The results of this study showed that the PCR-ELISA was sensitive and easy to perform without the use of any expensive and sophisticated equipment, the reagents used are easy to obtain commercially and advantageous over multiplex PCR since more than one type-specific probe is used and the selection of probes is more flexible

Potent Plasmablast-Derived Antibodies Elicited by the National Institutes of Health Dengue Vaccine

Exposure to dengue virus (DENV) is thought to elicit lifelong immunity, mediated by DENV-neutralizing antibodies (nAbs). However, Abs generated by primary infections confer serotype-specific protection, and immunity against other serotypes develops only after subsequent infections. Accordingly, the induction of these nAb responses acquired after serial DENV infections has been a long-sought-after goal for vaccination. Nonetheless, it is still unclear if tetravalent vaccines can elicit or recall nAbs. In this study, we have characterized the responses from a volunteer who had been previously exposed to DENV and was immunized with the live attenuated tetravalent vaccine Butantan-DV, developed by the NIH and Butantan Institute. Eleven days after vaccination, we observed an ∼70-fold expansion of the plasmablast population. We generated 21 monoclonal Abs (MAbs) from singly sorted plasmablasts. These MAbs were the result of clonal expansions and had significant levels of somatic hypermutation (SHM). Nineteen MAbs (90.5%) neutralized at least one DENV serotype at concentrations of 1 μg/ml or less; 6 of the 21 MAbs neutralized three or more serotypes. Despite the tetravalent composition of the vaccine, we observed a neutralization bias in the induced repertoire: DENV3 was targeted by 18 of the 19 neutralizing MAbs (nMAbs). Furthermore, the P3D05 nMAb neutralized DENV3 with extraordinary potency (concentration to achieve half-maximal neutralization [Neut ] = 0.03 μg/ml). Thus, the Butantan-DV vaccine engendered a mature, antigen-selected B cell repertoire. Our results suggest that preexisting responses elicited by a previous DENV3 infection were recalled by immunization. The dengue epidemic presents a global public health challenge that causes widespread economic burden and remains largely unchecked by existing control strategies. Successful control of the dengue epidemic will require effective prophylactic and therapeutic interventions. Several vaccine clinical efficacy trials are approaching completion, and the chances that one or more live attenuated tetravalent vaccines (LATVs) will be introduced worldwide is higher than ever. While it is widely accepted that dengue virus (DENV)-neutralizing antibody (nAb) titers are associated with protection, the Ab repertoire induced by LATVs remain uncharacterized. Here, we describe the isolation of potent (Neut < 0.1 μg/ml) nAbs from a DENV-seropositive volunteer immunized with the tetravalent vaccine Butantan-DV, which is currently in phase III trials