Comparative analysis of SARS-CoV-2 variants Alpha (B.1.1.7), Gamma (P.1), Zeta (P.2) and Delta (B.1.617.2) in Vero-E6 cells: ultrastructural characterization of cytopathology and replication kinetics

Abstract This study compares the effects of virus-cell interactions among SARS-CoV-2 variants of concern (VOCs) isolated in Brazil in 2021, hypothesizing a correlation between cellular alterations and mortality and between viral load and transmissibility. For this purpose, reference isolates of Alpha, Gamma, Zeta, and Delta variants were inoculated into monolayers of Vero-E6 cells. Viral RNA was quantified in cell supernatants by RT‒PCR, and infected cells were analyzed by Transmission Electron Microscopy (TEM) for qualitative and quantitative evaluation of cellular changes 24, 48, and 72 hours postinfection (hpi). Ultrastructural analyses showed that all variants of SARS-CoV-2 altered the structure and function of mitochondria, nucleus, and rough endoplasmic reticulum of cells. Monolayers infected with the Delta variant showed the highest number of modified cells and the greatest statistically significant differences compared to those of other variants. Viral particles were observed in the cytosol and the cell membrane in 100 % of the cells at 48 hpi. Alpha showed the highest mean particle diameter (79 nm), and Gamma and Delta were the smallest (75 nm). Alpha and Gamma had the highest particle frequency per field at 48 hpi, while the same was observed for Zeta and Delta at 72 hpi and 24 hpi, respectively. The cycle threshold of viral RNA varied among the target protein, VOC, and time of infection. The findings presented here demonstrate that all four VOCs evaluated caused ultrastructural changes in Vero-E6 cells, which were more prominent when infection occured with the Delta variant.

A case report of vaccine-induced immune thrombotic thrombocytopenia (VITT) with genetic analysis.

The emergence of the rare syndrome called vaccine-induced immune thrombocytopenia and thrombosis (VITT) after adenoviral vector vaccines, including ChAdOx1 nCov-19, raises concern about one's predisposing risk factors. Here we report the case of a 56-year-old white man who developed VITT leading to death within 9 days of symptom onset. He presented with superior sagittal sinus thrombosis, right frontal intraparenchymal hematoma, frontoparietal subarachnoid and massive ventricular hemorrhage, and right lower extremity arterial and venous thrombosis. His laboratory results showed elevated D-dimer, C-reactive protein, tissue factor, P-selectin (CD62p), and positive anti-platelet factor 4. The patient's plasma promoted higher CD62p expression in healthy donors' platelets than the controls. Genetic investigation on coagulation, thrombophilia, inflammation, and type I interferon-related genes was performed. From rare variants in European or African genomic databases, 68 single-nucleotide polymorphisms (SNPs) in one allele and 11 in two alleles from common SNPs were found in the patient genome. This report highlights the possible relationship between VITT and genetic variants. Additional investigations regarding the genetic predisposition of VITT are needed.

Long COVID-19 syndrome associated with Omicron XBB.1.5 infection: a case report

BACKGROUND There is interest in lingering non-specific symptoms after severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) infection, referred to as Long coronavirus disease 2019 (Long COVID-19). It remains unknown whether the risk of Long COVID-19 is associated with pre-existing comorbidities or initial COVID-19 severity, including infections due to new Omicron lineages which predominated in 2023. OBJECTIVES The aim of this case report was to characterize the clinical features of acute XBB.1.5 infection followed by Long COVID-19. METHODS We followed a 73-year old female resident of Rio de Janeiro with laboratory-confirmed SARS-CoV-2 during acute infection and subsequent months. The SARS-CoV-2 lineage was determined by genome sequencing. FINDINGS The participant denied comorbidities and had completed a two-dose vaccination schedule followed by two booster doses eight months prior to SARS-CoV-2 infection. Primary infection by viral lineage XBB.1.5. was clinically mild, but the participant subsequently reported persistent fatigue. MAIN CONCLUSIONS This case demonstrates that Long COVID-19 may develop even after mild disease due to SARS-CoV-2 in fully vaccinated and boosted individuals without comorbidities. Continued monitoring of new SARS-CoV-2 lineages and associated clinical outcomes is warranted. Measures to prevent infection should continue to be implemented including development of new vaccines and antivirals effective against novel variants.

Isolated intracranial hypertension associated with COVID-19.

BACKGROUND: Headache is a frequent complaint in COVID-19 patients. However, no detailed information on headache characteristics is provided in these reports. Our objective is to describe the characteristics of headache and the cerebrospinal fluid (CSF) profile in COVID-19 patients, highlighting the cases of isolated intracranial hypertension. METHODS: In this cross-sectional study, we selected COVID-19 patients who underwent lumbar puncture due to neurological complaints from April to May 2020. We reviewed clinical, imaging, and laboratory data of patients with refractory headache in the absence of other encephalitic or meningitic features. CSF opening pressures higher than 250 mmH2O were considered elevated, and from 200 to 250 mmH2O equivocal. RESULTS: Fifty-six COVID-19 patients underwent lumbar puncture for different neurological conditions. A new, persistent headache that prompted a CSF analysis was diagnosed in 13 (23.2%). The pain was throbbing, holocranial or bilateral in the majority of patients. All patients had normal CSF analysis and RT-qPCR for SARS-CoV-2 was negative in all samples. Opening pressure >200 mmH2O was present in 11 patients and, in six of these, > 250 mmH2O. 6/13 patients had complete improvement of the pain, five had partial improvement, and two were left with a daily persistent headache. CONCLUSIONS: In a significant proportion of COVID-19 patients, headache was associated to intracranial hypertension in the absence of meningitic or encephalitic features. Coagulopathy associated with COVID-19 could be an explanation, but further studies including post-mortem analysis of areas of production and CSF absorption (choroid plexuses and arachnoid granulations) are necessary to clarify this issue.

Atazanavir, Alone or in Combination with Ritonavir, Inhibits SARS-CoV-2 Replication and Proinflammatory Cytokine Production.

Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) is already responsible for far more deaths than previous pathogenic coronaviruses (CoVs) from 2002 and 2012. The identification of clinically approved drugs to be repurposed to combat 2019 CoV disease (COVID-19) would allow the rapid implementation of potentially life-saving procedures. The major protease (Mpro) of SARS-CoV-2 is considered a promising target, based on previous results from related CoVs with lopinavir (LPV), an HIV protease inhibitor. However, limited evidence exists for other clinically approved antiretroviral protease inhibitors. Extensive use of atazanavir (ATV) as antiretroviral and previous evidence suggesting its bioavailability within the respiratory tract prompted us to study this molecule against SARS-CoV-2. Our results show that ATV docks in the active site of SARS-CoV-2 Mpro with greater strength than LPV, blocking Mpro activity. We confirmed that ATV inhibits SARS-CoV-2 replication, alone or in combination with ritonavir (RTV) in Vero cells and a human pulmonary epithelial cell line. ATV/RTV also impaired virus-induced enhancement of interleukin 6 (IL-6) and tumor necrosis factor alpha (TNF-α) levels. Together, our data strongly suggest that ATV and ATV/RTV should be considered among the candidate repurposed drugs undergoing clinical trials in the fight against COVID-19.

Phosphatidyl Inositol 3 Kinase-Gamma Balances Antiviral and Inflammatory Responses During Influenza A H1N1 Infection: From Murine Model to Genetic Association in Patients.

Influenza A virus (IAV) infection causes severe pulmonary disease characterized by intense leukocyte infiltration. Phosphoinositide-3 kinases (PI3Ks) are central signaling enzymes, involved in cell growth, survival, and migration. Class IB PI3K or phosphatidyl inositol 3 kinase-gamma (PI3Kγ), mainly expressed by leukocytes, is involved in cell migration during inflammation. Here, we investigated the contribution of PI3Kγ for the inflammatory and antiviral responses to IAV. PI3Kγ knockout (KO) mice were highly susceptible to lethality following infection with influenza A/WSN/33 H1N1. In the early time points of infection, infiltration of neutrophils was higher than WT mice whereas type-I and type-III IFN expression and p38 activation were reduced in PI3Kγ KO mice resulting in higher viral loads when compared with WT mice. Blockade of p38 in WT macrophages infected with IAV reduced levels of interferon-stimulated gene 15 protein to those induced in PI3Kγ KO macrophages, suggesting that p38 is downstream of antiviral responses mediated by PI3Kγ. PI3Kγ KO-derived fibroblasts or macrophages showed reduced type-I IFN transcription and altered pro-inflammatory cytokines suggesting a cell autonomous imbalance between inflammatory and antiviral responses. Seven days after IAV infection, there were reduced infiltration of natural killer cells and CD8+ T lymphocytes, increased concentration of inflammatory cytokines in bronchoalveolar fluid, reduced numbers of resolving macrophages, and IL-10 levels in PI3Kγ KO. This imbalanced environment in PI3Kγ KO-infected mice culminated in enhanced lung neutrophil infiltration, reactive oxygen species release, and lung damage that together with the increased viral loads, contributed to higher mortality in PI3Kγ KO mice compared with WT mice. In humans, we tested the genetic association of disease severity in influenza A/H1N1pdm09-infected patients with three potentially functional PIK3CG single-nucleotide polymorphisms (SNPs), rs1129293, rs17847825, and rs2230460. We observed that SNPs rs17847825 and rs2230460 (A and T alleles, respectively) were significantly associated with protection from severe disease using the recessive model in patients infected with influenza A(H1N1)pdm09. Altogether, our results suggest that PI3Kγ is crucial in balancing antiviral and inflammatory responses to IAV infection.

Molecular diversity of human parvovirus B19 during two outbreaks of erythema infectiosum in Brazil

Abstract This study was conducted to provide information on the genetic diversity of human parvovirus B19 (B19V) circulating in the municipality of Niterói, Rio de Janeiro, Southeast Brazil during 1996-2006, a period with two distinct outbreaks of B19V infection: 1999-2000 and 2004-2005. A total of 27 sera from patients with erythema infectiosum and five sera from HIV-infected patients that tested positive for B19V DNA during the study period were analyzed. To genotype B19V strains, a semi-nested PCR for partial amplification of the capsid gene was performed and sequence analysis revealed that 31 sequences belonged to subgenotype 1a (G1a) of the main genotype 1 and one sequence was characterized as subgenotype 3b (G3b). The phylogenetic tree supported the division of the G1a into two well-defined clades with 1.3% of divergence. The low diversity of the G1a strains may be explained by the fact that all patients had acute B19V infection and 30/32 sera were collected during two distinct outbreaks. The G3b strain was from an HIV-infected patient who seroconverted to anti-B19 IgG antibodies in September/2005. This is the first report of G3b in the state of Rio de Janeiro.

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. (AU)

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.

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.

HIV-1 and its gp120 inhibits the influenza A(H1N1)pdm09 life cycle in an IFITM3-dependent fashion.

HIV-1-infected patients co-infected with A(H1N1)pdm09 surprisingly presented benign clinical outcome. The knowledge that HIV-1 changes the host homeostatic equilibrium, which may favor the patient resistance to some co-pathogens, prompted us to investigate whether HIV-1 infection could influence A(H1N1)pdm09 life cycle in vitro. We show here that exposure of A(H1N1)pdm09-infected epithelial cells to HIV-1 viral particles or its gp120 enhanced by 25% the IFITM3 content, resulting in a decrease in influenza replication. This event was dependent on toll-like receptor 2 and 4. Moreover, knockdown of IFITM3 prevented HIV-1 ability to inhibit A(H1N1)pdm09 replication. HIV-1 infection also increased IFITM3 levels in human primary macrophages by almost 100%. Consequently, the arrival of influenza ribonucleoproteins (RNPs) to nucleus of macrophages was inhibited, as evaluated by different approaches. Reduction of influenza RNPs entry into the nucleus tolled A(H1N1)pdm09 life cycle in macrophages earlier than usual, limiting influenza's ability to induce TNF-α. As judged by analysis of the influenza hemagglutin (HA) gene from in vitro experiments and from samples of HIV-1/A(H1N1)pdm09 co-infected individuals, the HIV-1-induced reduction of influenza replication resulted in delayed viral evolution. Our results may provide insights on the mechanisms that may have attenuated the clinical course of Influenza in HIV-1/A(H1N1)pdm09 co-infected patients during the recent influenza form 2009/2010.

Detection of the influenza A(H1N1)pdm09 virus carrying the K-15E, P83S and Q293H mutations in patients who have undergone bone marrow transplant.

The 2009 pandemic influenza A(H1N1)pdm09 virus emerged and caused considerable morbidity and mortality in the third world, especially in Brazil. Although circulating strains of A(H1N1)pdm09 are A/California/04/2009-like (CA-04-like) viruses, various studies have suggested that some mutations in the viral hemagglutinin (HA) may be associated with enhanced severity and fatality. This phenomenon is particularly challenging for immunocompromised individuals, such as those who have undergone bone marrow transplant (BMT), because they are more likely to display worse clinical outcomes to influenza infection than non-immunocompromised individuals. We studied the clinical and viral aspects of post-BMT patients with confirmed A(H1N1)pdm09 diagnosis in the largest cancer hospital in Brazil. We found a viral strain with K-15E, P83S and Q293H polymorphisms in the HA, which is presumably more virulent, in these individuals. Despite that, these patients showed only mild symptoms of infection. Our findings complement the discovery of mild cases of infection with the A(H1N1)pdm09 virus with the K-15E, P83S and Q293H mutations in Brazil and oppose other studies that have linked these changes with increased disease severity. These results could be important for a better comprehension of the impact of the pandemic influenza in the context of BMT.

Respiratory syncytial virus epidemic periods in an equatorial city of Brazil.

BACKGROUND: Characterization of the human respiratory syncytial virus (HRSV) season at the local level has important implications for appropriate decisions on the time period for administration of specific prophylaxis. OBJECTIVES: (1) To describe five consecutive epidemic periods of HRSV in an equatorial city of Brazil and (2) to show preliminary data on genomic diversity of circulating HRSV. PATIENTS/METHODS: Nasopharyngeal aspirates of 2885 children attending the emergency room and wards of a public hospital were collected and screened by indirect immunofluorescence for HRSV infections during five consecutive years (from January 2004 to December 2008). In addition, the genetic and antigenic variability of the HRSV strains isolated was evaluated by partial nucleotide sequencing of the protein G gene. RESULTS: HRSV was detected in 15·8% of the analyzed samples. HRSV seasons occurred in a restricted period of each year. The onset of each HRSV season was variable (February to May), but the end always occurred in July. From the 456 HRSV infections found, 86 cases with bronchiolitis were genotyped. Both HRSV subgroups (A and B) cocirculated during the five epidemic periods. The 58 HRSV-A strains grouped into two clades, GA2 and GA5. In respect of the HRSV-B strains, the 28 samples grouped into two clades: GB3 and BA. CONCLUSIONS: HRSV accounts for a substantial proportion of ARI in the study population. As in temperate countries, HRSV infections in this equatorial area of Brazil also cause seasonal yearly epidemics, and this has implications for prophylaxis strategies. The city of Fortaleza follows the same worldwide trend of circulation of genotypes of HRSV.

H1N1pdm influenza infection in hospitalized cancer patients: clinical evolution and viral analysis.

BACKGROUND: The novel influenza A pandemic virus (H1N1pdm) caused considerable morbidity and mortality worldwide in 2009. The aim of the present study was to evaluate the clinical course, duration of viral shedding, H1N1pdm evolution and emergence of antiviral resistance in hospitalized cancer patients with severe H1N1pdm infections during the winter of 2009 in Brazil. METHODS: We performed a prospective single-center cohort study in a cancer center in Rio de Janeiro, Brazil. Hospitalized patients with cancer and a confirmed diagnosis of influenza A H1N1pdm were evaluated. The main outcome measures in this study were in-hospital mortality, duration of viral shedding, viral persistence and both functional and molecular analyses of H1N1pdm susceptibility to oseltamivir. RESULTS: A total of 44 hospitalized patients with suspected influenza-like illness were screened. A total of 24 had diagnosed H1N1pdm infections. The overall hospital mortality in our cohort was 21%. Thirteen (54%) patients required intensive care. The median age of the studied cohort was 14.5 years (3-69 years). Eighteen (75%) patients had received chemotherapy in the previous month, and 14 were neutropenic at the onset of influenza. A total of 10 patients were evaluated for their duration of viral shedding, and 5 (50%) displayed prolonged viral shedding (median 23, range=11-63 days); however, this was not associated with the emergence of a resistant H1N1pdm virus. Viral evolution was observed in sequentially collected samples. CONCLUSIONS: Prolonged influenza A H1N1pdm shedding was observed in cancer patients. However, oseltamivir resistance was not detected. Taken together, our data suggest that severely ill cancer patients may constitute a pandemic virus reservoir with major implications for viral propagation.

The epidemiology and antigenic characterization of influenza viruses isolated in Curitiba, South Brazil

Several studies conducted all over the world have reported that the influenza virus is associated with great morbidity and mortality rates. In this study, we analyzed the incidence of the influenza virus between 2000 and 2003 in Curitiba. We studied 1621 samples obtained from outpatients and hospitalized patients of both sexes and all ages. The study was conducted at the local primary care health units (outpatients) and at the tertiary care unit (hospitalized) of the General Hospital of the Federal University in the state of Paraná, Brazil. Nasopharyngeal aspirates and, eventually, bronchoalveolar lavage were assayed for the presence of viral antigens, either by indirect immunofluorescence or cell culture. Of the samples studied, 135 (8.3 percent) were positive for influenza virus, and of those, 103 (76.3 percent) were positive for type A and 32 (23.7 percent) for type B. Additionally, positive samples were analyzed by reverse transcription followed by polymerase chain reaction and subtypes H1 and H3 were identified from this group. A high incidence of positive samples was observed mainly in the months with lower temperatures. Furthermore, outpatients showed a higher incidence of influenza viruses than hospitalized patients.

Antigenic and genomic characterization of adenovirus associated to respiratory infections in children living in Northeast Brazil

From January to December 1998, nasopharyngeal aspirates were obtained from 482 children with acute respiratory infections attended in emergence department and wards of a teaching hospital in the city of Salvador, Brazil. The samples were tested for the presence of adenovirus by isolation in tissue culture and indirect immunofluorescence assay. Eleven adenoviruses were detected by both methods in the same clinical samples. Infections by adenovirus were observed during seven months of the year without association with rainy season. Genome analysis was performed on these 11 isolates. Species C was represented by serotypes 1, 2 and 5. Within species B, only serotype 7 (Ad7) was detected. Two genomic variants of Ad1, two variants of Ad2, one of Ad5, and one of Ad7 (7h) were identified. This is the first study of molecular epidemiology of adenovirus associated to acute respiratory infections in children living in Northeast Brazil, and contributes to a better understanding of adenovirus infections in the country.

A rubella serosurvey in postpartum women in the three regions of Peru

OBJECTIVE: To determine the proportion of postpartum women aged 15-49 in Peru who are susceptible to rubella, in order to help address strategies to eliminate rubella and to prevent congenital rubella syndrome (CRS) in the country. METHODS: A cross-sectional survey was conducted during March and April 2003 in six main regional hospitals, in the three geographic regions (coast, mountain, and jungle) of Peru. For the postpartum women who provided written informed consent, a questionnaire was administered and a blood specimen was collected. Sera were tested for rubella immunoglobulin G (IgG) antibody, using a commercial enzyme-linked immunosorbent serologic assay (ELISA) kit. Univariate, bivariate, and multivariate analyses were carried out to assess risk factors for susceptibility. RESULTS: In total, 1 236 postpartum women were enrolled. The overall proportion of IgG-antibody negative women was 12.8 percent (95 percent confidence interval (CI): 10.9 percent-14.6 percent). Bivariate analysis found the following variables associated with susceptibility: living in the jungle region (odds ratio (OR) = 1.65; 95 percent CI: 1.13-2.42); age < 19 years (OR = 2.02; 95 percent CI: 1.35-3.03); being a housewife (OR = 1.69; 95 percent CI: 1.12-2.55); and having < 11 years of education (OR = 2.12; 95 percent CI :1.20-3.75). Multivariate analysis found the following variables were associated with susceptibility: living in the jungle region (OR = 1.67; 95 percent CI: 1.13-2.46); age < 19 years (OR = 1.62; 95 percent CI: 1.07-2.47); having < 4 children born alive (OR = 1.85; 95 percent CI: 1.00- 3.40); and having < 11 years of education (OR = 2.07; 95 percent CI: 1.16-3.71). CONCLUSIONS: The proportion of postpartum women at the study sites who were found to be susceptible to rubella was 12.8 percent, placing Peru among the countries facing a moderate level of risk for the occurrence of CRS cases. The findings suggest the need to also provide...

OBJETIVO: Determinar la proporción de mujeres recién paridas de 15-49 años de edad susceptibles a la rubéola en Perú, a fin de contribuir a establecer estrategias para eliminar la rubéola y evitar el síndrome de rubéola congénita (SRC) en el país. MÉTODOS: Se realizó un estudio transversal en marzo y abril de 2003 en seis hospitales regionales principales de tres regiones geográficas (costa, sierra y selva) de Perú. A las mujeres recién paridas que dieron su consentimiento informado por escrito se les aplicó un cuestionario y se les tomó una muestra de sangre. La detección serológica de anticuerpos de la clase IgG contra rubéola se realizó mediante un ensayo inmunoenzimático comercial tipo ELISA. Se realizaron análisis con una, dos y múltiples variables para evaluar los factores de riesgo de ser susceptible a la enfermedad. RESULTADOS: En total participaron 1 236 mujeres recién paridas. La proporción de mujeres negativas a anticuerpos IgG contra rubéola fue de 12,8 por ciento (intervalo de confianza de 95 por ciento [IC95 por ciento]: 10,9 por ciento a 14,6 por ciento). Según el análisis bifactorial, las variables asociadas con la vulnerabilidad a la enfermedad fueron: vivir en la región de la selva (razón de posibilidades [odds ratio, OR] = 1,65; IC95 por ciento: 1,13 a 2,42); tener menor de 19 años de edad (OR = 2,02; IC95 por ciento: 1,35 a 3,03); ser ama de casa (OR = 1,69; IC95 por ciento: 1,12 a 2,55); y tener 11 años o menos de educación (OR = 2,12; IC95 por ciento: 1,20 a 3,75). Las variables asociadas con la vulnerabilidad a la enfermedad según el análisis multifactorial fueron: vivir en la región de la selva (OR = 1,67; IC95 por ciento: 1,13 a 2,46); tener menos de 19 años de edad (OR = 1,62; IC95 por ciento: 1,07 a 2,47); tener menos de 4 niños nacidos vivos (OR = 1,85; IC95 por ciento: 1,00 a 3,40); y tener 11 años o menos de educación (OR = 2,07; IC95 por ciento: 1,16 a 3,71). CONCLUSIONES: En la zona estudiada, la proporción...

Antigenic and genomic characterization of adenovirus associated to respiratory infections in children living in Northeast Brazil.

From January to December 1998, nasopharyngeal aspirates were obtained from 482 children with acute respiratory infections attended in emergence department and wards of a teaching hospital in the city of Salvador, Brazil. The samples were tested for the presence of adenovirus by isolation in tissue culture and indirect immunofluorescence assay. Eleven adenoviruses were detected by both methods in the same clinical samples. Infections by adenovirus were observed during seven months of the year without association with rainy season. Genome analysis was performed on these 11 isolates. Species C was represented by serotypes 1, 2 and 5. Within species B, only serotype 7 (Ad7) was detected. Two genomic variants of Ad1, two variants of Ad2, one of Ad5, and one of Ad7 (7h) were identified. This is the first study of molecular epidemiology of adenovirus associated to acute respiratory infections in children living in Northeast Brazil, and contributes to a better understanding of adenovirus infections in the country.

Prevalence of antibodies against measles, mumps, and rubella before and after vaccination of school-age children with three different triple combined viral vaccines, Rio Grande do Sul, Brazil, 1996

OBJECTIVE: We evaluated the seroprevalence for measles, mumps, and rubella in school-age children (6-12 years old) before and after the administration of three triple combined viral vaccines. METHODS: In two municipal schools of Rio Grande do Sul, Brazil, 692 blood samples were collected before vaccination and 636 samples 21 to 30 days after vaccination during 1996. IgG antibody seropositivity was investigated by enzyme-linked immunosorbent assay (measles and mumps with Enzygnost [Behring, Marburg, Germany]; rubella with Rubenostika [Organon Teknica, Boxtel, the Netherlands]). The vaccines compared were: A: E-Zagreb, L-Zagreb, and Wistar RA 27/3 (Tresivac); B: Moraten, J-Lynn, and Wistar RA 27/3 (M-M-R II); and C: Schwarz, Urabe AM-9, and Wistar RA 27/3 (Trimovax). RESULTS: Before vaccination, 79.2 percent [95 percent confidence interval (CI) = 76.0 percent-82.2 percent] of the samples were positive for measles, 69.4 percent (95 percent CI = 65.8 percent-72.8 percent) for mumps, and 55.4 percent (95 percent CI = 51.6 percent-59.2 percent) for rubella. After vaccination with the A, B, and C vaccines, seropositivity was 100.0 percent, 99.5 percent, and 100.0 percent, respectively for measles; 99.5 percent, 94.5 percent, and 92.0 percent for mumps; and 92.6 percent, 91.3 percent, and 88.6 percent for rubella. CONCLUSIONS: About one-fifth (20.8 percent) of the schoolchildren who could have been vaccinated against measles at age 9 months had levels of antibodies insufficient for protection. In the sample of schoolchildren without previous vaccination against mumps and rubella, high proportions of susceptible levels were found. All vaccines were immunogenic, but vaccine A yielded a seroconversion rate of 99.5 percent for the mumps component, which was significantly higher than the other two vaccines (P < 0.01).

OBJETIVO: Se evaluó la seroprevalencia para sarampión, paperas y rubéola en niños en edad escolar (6-12 años) antes y después de la administración de tres vacunas triples antivirales combinadas. MÉTODOS: Se colectaron 692 muestras de sangre antes de la vacunación y 636 muestras entre 21 y 30 días después de la vacunación a niños de dos escuelas municipales de Rio Grande do Sul, Brasil, durante 1996. Se investigó la seropositividad de anticuerpos de la clase IgG mediante un ensayo de inmunoadsorción enzimática tipo ELISA (sarampión y paperas con Enzygnost [Behring, Marburgo, Alemania] y rubéola con Rubenostika [Organon Teknica, Boxtel, Países Bajos]). Las vacunas comparadas fueron: a) E-Zagreb, L-Zagreb y Wistar RA 27/3 (Tresivac); b) Moraten, J-Lynn y Wistar RA 27/3 (M-M-R II); y c) Schwarz, Urabe AM-9 y Wistar RA 27/3 (Trimovax). RESULTADOS: Antes de la vacunación, 79,2 por ciento (intervalo de confianza [IC] 95 por ciento: 76,0 a 82,2) de las muestras fueron positivas para sarampión, 69,4 por ciento (IC 95 por ciento: 65,8 a 72,8) para paperas y 55,4 por ciento (IC 95 por ciento: 51,6 a 59,2) para rubéola. Después de la vacunación con las vacunas A, B y C, la seropositividad fue de 100 por ciento, 99,5 por ciento y 100 por ciento, respectivamente para sarampión; de 99,5 por ciento, 94,5 por ciento y 92,0 por ciento para paperas; y de 92,6 por ciento, 92,3 por ciento y 88,6 por ciento para rubéola. CONCLUSIONES: Alrededor de un quinto (20,8 por ciento) de los escolares que pudieron haber sido vacunados contra el sarampión a los 9 meses de edad tenían niveles de anticuerpos insuficientes para protegerlos. En la muestra de escolares sin vacunación previa contra paperas y rubéola se encontró una alta proporción de niños susceptibles. Todas las vacunas fueron inmunogénicas, pero la vacuna A produjo una tasa de seroconversión de 99,5 por ciento para el componente de paperas, significativamente mayor que la de las otras dos vacunas (P < 0,01).

Etiologia de exantema em crianças em uma área endêmica de dengue / Etiology of exanthema in children in a dengue endemic area

OBJETIVO: Estudar a etiologia dos casos de exantema com ou sem febre em crianças atendidas no pronto-socorro de um hospital de uma zona endêmica para dengue. MÉTODOS: No período de 21/09/2001 a 20/09/2002, foram inscritas no estudo 95,9 por cento (71/74) das crianças atendidas no pronto-socorro do Hospital Universitário de Campo Grande (MS) que apresentassem exantema (percentual de recusa de 4,1 por cento). Após preenchimento do protocolo com os dados das crianças, as mesmas foram submetidas a exame físico seguido da coleta de amostras de sangue para realizar hemograma com contagem de plaquetas e sorologias (IgM e IgG); inicialmente para dengue, rubéola e toxoplasmose e, posteriormente, naqueles casos com resultado negativo, realizou-se sorologia para parvovirose, herpes vírus tipo 6 e sarampo. RESULTADOS: O diagnóstico laboratorial foi confirmado através da pesquisa de anticorpo IgM em 88,7 por cento dos casos investigados: dengue (77,5 por cento), herpes vírus tipo 6 (8,4 por cento), parvovirose (2,8 por cento) e diagnóstico inconclusivo em oito pacientes (11,3 por cento). Não foi evidenciada sorologia positiva (IgM) para sarampo, rubéola ou toxoplasmose naquela ocasião. As manifestações clínicas mais freqüentes nos pacientes com dengue foram: febre, prurido, prostração, mialgia e prova do laço positiva. Nos pacientes cujo diagnóstico foi dengue, a prova do laço foi positiva em 58,4 por cento (32/55) dos casos, demonstrando diferença estatisticamente significativa quando comparada com o grupo cujo diagnóstico não foi dengue. CONCLUSÕES: Nas crianças com exantema, dengue pode ser a principal enfermidade causal, atentando-se para a epidemiologia do local. É necessário um controle constante da vigilância epidemiológica e sorológica das doenças exantemáticas.

OBJECTIVE: To study the etiology of exanthema cases, with or without fever, in children seen in the emergency room of a hospital located in a region where dengue is endemic. METHODS: Enrollment took place between 21/09/2001 and 20/09/2002 and included 95.9 percent (71/74) of children presenting with exanthema at the emergency room of the Hospital Universitário de Campo Grande, MS (4.1 percent refusals). After the children had had their details taken and entered on the study protocol, they were subjected to physical examination followed by collection of blood samples for blood testing with platelet counts and serology (IgM and IgG); initially for dengue, rubella and toxoplasmosis and then, in negative cases, serology was also run for parvovirus, herpes virus type 6 and measles. RESULTS: Laboratory diagnoses were confirmed by means of IgM antibody assay in 88.7 percent of the cases investigated: dengue (77.5 percent), herpes virus type 6 (8.4 percent), parvovirus (2.8 percent) and in eight patients diagnosis was inconclusive (11.3 percent). On this occasion no positive serology (IgM) was observed for measles, rubella or toxoplasmosis. The most common clinical manifestations among the dengue patients were: fever, itching, prostration, myalgia and positive tourniquet test results. In 58.4 percent (32/55) of those cases diagnosed with dengue, the tourniquet test was positive, which was a statistically significant difference when compared with the remainder of the sample. CONCLUSIONS: When children present with exanthema, it is possible that dengue is the primary causative disease, depending on the epidemiology of the location. Constant control of epidemiological and serological surveillance of exanthematous diseases is necessary.