Influenza virus A(H3N2) strain isolated from cerebrospinal fluid from a patient presenting myelopathy post infectious.

BACKGROUND: Neurological involvement during influenza infection has been described during epidemics and is often consistent with serious sequelae or death. OBJECTIVE: To investigate the etiologic agent involved in myelopathy post influenza-like syndrome. STUDY DESIGN: This investigation focuses on virus isolation from the cerebrospinal fluid (CSF) collected from a 19-year-old male student presenting with clinical diagnosis of myelopathy post influenza-like syndrome. To achieve this goal, different cell cultures and molecular methodologies were carried out. RESULTS: Influenza virus A(H3N2) strain was isolated in MDCK cell culture; virus particles were observed under electron microscopy. Phylogenetics analyses showed that the Brazilian influenza A(H3N2) strains were closely related to the A/Perth/16/2009-like. CONCLUSION: This study demonstrates that influenza virus A(H3N2) strain was the cause of illness of the students. According to the Brazilian influenza virus sentinel surveillance data A/Perth/16/2009-LIKE (H3N2) strain has predominated during the 2010 influenza virus season in Brasília-DF.

ESTUDO RETROSPECTIVO DE 1.137 ANIMAIS SUBMETIDOS À ACUPUNTURA NA FMVZ - UNESP - BOTUCATU - SP.

As evidências da eficácia e da relação custo-benefício de uma terapia são obtidas com estudos primários descritivos, analíticos retrospectivos e prospectivos. Este trabalho mostra o levantamento retrospectivo de casos clínicos do ambulatório de Acupuntura Veterinária do Departamento de Cirurgia e Anestesiologia Veterinária (DCAV) . FMVZ . UNESP . Botucatu, durante o período de 1998 a 2009, totalizando 1.137 prontuários. Foram avaliados a eficácia da acupuntura, a evolução clínica, a necessidade ou não de terapêuticas complementares à acupuntura e o perfil do paciente atendido. Os pacientes atendidos apresentaram distúrbios comprometendo principalmente os sistemas neurológico e musculoesquelético. Esses pacientes são, em sua maioria, da espécie canina e portadores de doença crônica ou doenças de baixa resposta à terapêutica usual (alopatia). E essa prevalência provavelmente se deve à eficácia da acupuntura no tratamento de doença do disco intervertebral. A prevalência de alterações neurológicas pode se dever ao fato de que estes casos não responderam de forma satisfatória ao tratamento farmacológico. O baixo índice de adesão ao tratamento com acupuntura pode estar associado ao tipo de paciente encaminhado, em sua maioria os que não se beneficiaram do tratamento farmacológico ou cirúrgico preconizado. Quase a metade (47%) dos casos atendidos apresentaram como primeira opção a acu

Estudo retrospectivo de 1137 animais submetidos à acupuntura na FMVZ-UNESP-Botucatu-SP / Retrospective study of 1137 cases submmited to acupuncture at FMVZ-UNESP-Botucatu-SP

As evidências da eficácia e da relação custo-benefício de uma terapia são obtidas com estudos primários descritivos, analíticos retrospectivos e prospectivos. Este trabalho mostra o levantamento retrospectivo de casos clínicos do ambulatório de Acupuntura Veterinária do Departamento de Cirurgia e Anestesiologia Veterinária (DCAV) - FMVZ - UNESP - Botucatu, durante o período de 1998 a 2009, totalizando 1.137 prontuários. Foram avaliados a eficácia da acupuntura, a evolução clínica, a necessidade ou não de terapêuticas complementares à acupuntura e o perfil do paciente atendido. Os pacientes atendidos apresentaram distúrbios comprometendo principalmente os sistemas neurológico e musculoesquelético. Esses pacientes são, em sua maioria, da espécie canina e portadores de doença crônica ou doenças de baixa resposta à terapêutica usual (alopatia). E essa prevalência provavelmente se deve à eficácia da acupuntura no tratamento de doença do disco intervertebral. A prevalência de alterações neurológicas pode se dever ao fato de que estes casos não responderam de forma satisfatória ao tratamento farmacológico. O baixo índice de adesão ao tratamento com acupuntura pode estar associado ao tipo de paciente encaminhado, em sua maioria os que não se beneficiaram do tratamento farmacológico ou cirúrgico preconizado. Quase a metade (47%) dos casos atendidos apresentaram como primeira opção a acupuntura.

The evidences of efficiency and the relation of cost-benefit of therapy are obtained with revisions developed from primary studies, which includes descriptive, analytical, retrospective and prospective. This is a retrospective survey from the Acupuncture Clinic from the Faculty of Veterinary Medicine and Animal Science, Sao Paulo State University, Botucatu, Brazil from 1998 to 2009. Acupuncture efficacy, clinical evolution, the need or not of complementary therapies to acupuncture and the patient profile were evaluated in 1137 clinical charts. The main diseases presented by attended patients involve neurological and musculoskeletal problems. The majority of the patients are dogs and presents chronic diseases or low-response to usual therapies (allopathic). This prevalence is probably due to acupuncture efficacy in disc disease. The poor compliance to acupuncture treatment can be related the patient profile: animals that did not benefit from pharmacological or surgical recommended procedures. Acupuncture was the first choice for almost half (47%) of the patients.

ESTUDO RETROSPECTIVO DE 1.137 ANIMAIS SUBMETIDOS À ACUPUNTURA NA FMVZ - UNESP - BOTUCATU - SP.

As evidências da eficácia e da relação custo-benefício de uma terapia são obtidas com estudos primários descritivos, analíticos retrospectivos e prospectivos. Este trabalho mostra o levantamento retrospectivo de casos clínicos do ambulatório de Acupuntura Veterinária do Departamento de Cirurgia e Anestesiologia Veterinária (DCAV) . FMVZ . UNESP . Botucatu, durante o período de 1998 a 2009, totalizando 1.137 prontuários. Foram avaliados a eficácia da acupuntura, a evolução clínica, a necessidade ou não de terapêuticas complementares à acupuntura e o perfil do paciente atendido. Os pacientes atendidos apresentaram distúrbios comprometendo principalmente os sistemas neurológico e musculoesquelético. Esses pacientes são, em sua maioria, da espécie canina e portadores de doença crônica ou doenças de baixa resposta à terapêutica usual (alopatia). E essa prevalência provavelmente se deve à eficácia da acupuntura no tratamento de doença do disco intervertebral. A prevalência de alterações neurológicas pode se dever ao fato de que estes casos não responderam de forma satisfatória ao tratamento farmacológico. O baixo índice de adesão ao tratamento com acupuntura pode estar associado ao tipo de paciente encaminhado, em sua maioria os que não se beneficiaram do tratamento farmacológico ou cirúrgico preconizado. Quase a metade (47%) dos casos atendidos apresentaram como primeira opção a acu

Performance of indirect immunofluorescence assay, immunochromatography assay and reverse transcription-polymerase chain reaction for detecting human respiratory syncytial virus in nasopharyngeal aspirate samples.

Comparison of the use of indirect immunofluorescence assay (IFA), immunochromatography assay (ICA-BD) and reverse transcription-polymerase chain reaction (RT-PCR) for detecting human respiratory syncytial virus (HRSV) in 306 nasopharyngeal aspirates samples (NPA) was performed in order to assess their analytical performance. By comparing the results obtained using ICA-BD with those using IFA, we found relative indices of 85.0% for sensitivity and 91.2% for specificity, and the positive (PPV) and negative (NPV) predictive values were 85.0% and 91.2%, respectively. The relative indices for sensitivity and specificity as well as the PPV and NPV for RT-PCR were 98.0%, 89.0%, 84.0% and 99.0%, respectively, when compared to the results of IFA. In addition, comparison of the results of ICA-BD and those of RT-PCR yielded relative indices of 79.5% for sensitivity and 95.4% for specificity, as well as PPV and NPV of 92.9% and 86.0%, respectively. Although RT-PCR has shown the best performance, the substantial agreement between the ICA-BD and IFA results suggests that ICA-BD, also in addition to being a rapid and facile assay, could be suitable as an alternative diagnostic screening for HRSV infection in children.

Performance of indirect immunofluorescence assay, immunochromatography assay and reverse transcription-polymerase chain reaction for detecting human respiratory syncytial virus in nasopharyngeal aspirate samples

Comparison of the use of indirect immunofluorescence assay (IFA), immunochromatography assay (ICA-BD) and reverse transcription-polymerase chain reaction (RT-PCR) for detecting human respiratory syncytial virus (HRSV) in 306 nasopharyngeal aspirates samples (NPA) was performed in order to assess their analytical performance. By comparing the results obtained using ICA-BD with those using IFA, we found relative indices of 85.0 percent for sensitivity and 91.2 percent for specificity, and the positive (PPV) and negative (NPV) predictive values were 85.0 percent and 91.2 percent, respectively. The relative indices for sensitivity and specificity as well as the PPV and NPV for RT-PCR were 98.0 percent, 89.0 percent, 84.0 percent and 99.0 percent, respectively, when compared to the results of IFA. In addition, comparison of the results of ICA-BD and those of RT-PCR yielded relative indices of 79.5 percent for sensitivity and 95.4 percent for specificity, as well as PPV and NPV of 92.9 percent and 86.0 percent, respectively. Although RT-PCR has shown the best performance, the substantial agreement between the ICA-BD and IFA results suggests that ICA-BD, also in addition to being a rapid and facile assay, could be suitable as an alternative diagnostic screening for HRSV infection in children.

Identification of a bisegmented double-stranded RNA virus (picobirnavirus) in calf faeces.

To determine the incidence of rotavirus infection among dairy herds in the State of São Paulo, Brazil, 576 faecal samples obtained from calves aged 1-45 days with and without diarrhoea, reared on 63 dairy cattle farms, were analyzed. Polyacrylamide gel electrophoresis (PAGE) identified 28 samples positive for group A rotavirus, while four samples, two diarrhoeic and two non-diarrhoeic, showed a bisegmented genome with a typical picobirnavirus pattern. Electron microscopy revealed spherical virus particles with a diameter of 37 nm and without a defined surface structure. The present study is the first report of a bisegmented virus identified in cattle in Brazil.

HIV-1 diversity in Brazil: genetic, biologic, and immunologic characterization of HIV-1 strains in three potential HIV vaccine evaluation sites. Brazilian Network for HIV Isolation and Characterization.

The Brazilian Network for HIV Isolation and Characterization was established for the surveillance of HIV variability in Brazil. Here, we report characterization of HIV strains and virus-specific immune responses from 35 clinical samples collected from three potential HIV vaccine sites. Three genetic subtypes of HIV-1 were identified by heteroduplex mobility assay (HMA) B (in 82.9% of the samples), F (14.3%), and C (2.9%). Phylogenetic analysis based on the C2V3/env DNA sequence from all 25 specimens examined was 100% concordant with HMA results. Four variants of subtype B with different tetrapeptides at the tip of the V3 loop were found: the GPGR motif (North American), GWGR motif (Brazilian B"), and two minor variants, GFGR and GPGS, as previously detected. No significant association was found between HIV-1 subtypes and the mode of transmission or biologic properties of HIV-1 isolates (derived from 88.6% of the specimens). Only 5 of 16 isolates studied were neutralized by the autologous sera. Consistent with previous results, no relation between viral subtype and peptide enzyme-linked immunosorbent assay (ELISA) seroreactivity or neutralization was evident. This study also demonstrated the effectiveness of the collaborative approach followed by Brazilian scientists when addressing a complex subject such as HIV variability.

Analysis of Treponema pallidum recombinant antigens for diagnosis of syphilis by western blotting technique.

Three GST fusion recombinant antigen of Treponema pallidum, described as GST-rTp47, GST-rTp17 and GST-rTp15 were analyzed by Western blotting techniques. We have tested 53 serum samples: 25 from patients at different clinical stages of syphilis, all of them presenting anti-treponemal antibody, 25 from healthy blood donors and three from patients with sexually transmitted disease (STD) other than syphilis. Almost all samples from patients with syphilis presented a strong reactivity with GST-rTp17 antigen. Some samples were non-reactive or showed a weak reaction with GST-rTp47 and/or GST-rTp15, and apparently there was no correlation with the stage of disease. There was no seropositivity among blood donors. No sample reacted with purified GST. We concluded that due to their specificity these recombinant antigens can be used as GST fusion protein for development of syphilis diagnostic assays.

Analysis of cells in cerebrospinal fluid from patients with neurocysticercosis by means of flow cytometry.

The events of the cellular immune response in neurocysticercosis (NC) are not fully understood. Studies of the CD3, CD3/CD4, CD3/CD8, CD45/CD19, and CD45/CD56 molecules and activation-related CD69 molecule in cells from the cerebrospinal fluid (CSF) and peripheral blood (PB) of patients with NC may provide a better elucidation of the inflammatory and immunological events occurring in this disease. Seven patients with NC and 3 individuals with other disorders were evaluated by a three-color flow cytometric method. CD69 was detected in a higher percentage of cells in all CSF samples from patients, but not in PB or CSF from the control group. The percentage of CD3+ cells did not differ significantly in CSF and PB cells from patients and controls. The predominance of CD3+CD8+ cells was observed in CSF from one patient and in PB from 2 patients, who were in stage III of the disease (inflammatory process). The percentage of CD45+CD19+ cells was higher in CSF than in PB from patients who presented anti-cysticercus antibodies in CSF. The percentage of CD45+CD56+ cells in CSF was higher than in PB, but this rate was similar to reference values reported by other authors. Our data suggest that the cytometric method applied to a larger number of CSF samples may provide a better understanding of the cell-mediated immune response involved in NC.

An outbreak of group C rotavirus gastroenteritis among adults living in Valentim Gentil, São Paulo State, Brazil.

An outbreak of gastroenteritis affecting adults and children occurred in the small city of Valentim Gentil, São Paulo, Brazil, in 1993. Nineteen faecal samples (from 10 cases and 9 contracts) were examined by direct electron microscopy (DEM), immune electron microscopy (IEM), polyacrylamide gel electrophoresis (PAGE), and enzyme-linked immunosorbent assay (ELISA) for group A and C rotaviruses. DEM detected rotavirus in 6 of the 10 cases and in none of the contacts. All of the samples were negative for group A rotavirus by ELISA. Analysis by PAGE showed an electrophoretic profile suggestive of group C rotavirus in two cases. Group C rotavirus was identified by IEM in 4 of the cases and in 1 of the contacts. All of the samples were submitted to ELISA for group C rotavirus. This resulted in a total of 10 positives-7 for diarrhoeal cases and 3 for contacts. This outbreak was strongly associated with group C rotavirus. The importance of combining different diagnostic methods is emphasised.

Intrafamilial outbreak of astrovirus gastroenteritis in São Paulo, Brazil.

This report describes an intrafamilial outbreak of astrovirus-associated gastroenteritis that occurred among 5 children in a family living in the State of São Paul, Brazil. Astrovirus was detected by direct electron microscopy (DEM) and confirmed by solid phase immune electron microscopy (SPIEM) using astrovirus reference serum (T2-6-89). The rise in titre of astrovirus antibody in patients' sera was observed by IEM using acute and convalescent-phase sera. Combined use of the EM techniques to detect virus in stool and antibody rise in sera led us to conclude that this intrafamilial outbreak might have been caused by astrovirus infection. In Brazil, astrovirus may be a more common agent of gastroenteritis than in currently appreciated.

[Rotavirus, adenovirus, astrovirus, calicivirus and small round virus particles in feces of children with and without acute diarrhea, from 1987 to 1988, in the greater São Paulo]. / Rotavírus, adenovírus, astrovírus, calicivírus e "small round virus particles" em fezes de crianças, com e sem diarréia aguda, no período de 1987 a 1988, na grande São Paulo.

Between 1987 and 1988, 193 faecal specimens from children, with or without diarrhea, were submitted to enzyme immunoassay, polyacrylamide-gel electrophoresis and electronmicroscopy tests for virus detection. The positivity for Rotavirus, Adenovirus, Astrovirus, Calicivirus and Small Round Virus Particles (SRVP) was 11.3%, 3.1%, 2.1%, 1.0% and 4.1%, respectively, for the 97 children with acute diarrhea. Of the 96 children without diarrhea, 4.2% were positive for Rotavirus, 1.0% for Calicivirus and 7.3% for SRVP. Of 15 positive specimens for Rotavirus, 14 showed electrophoretic patterns proper to group A and 1 specimen of group C Rotavirus. The analysis of electrophorotypes demonstrated great heterogeneity of electrophoretic patterns and predominance of subgroup 2, "long". The association of virus, bacteria and parasites was present both in children with or without acute diarrhea.

Preliminary characterization of a new virus isolated from the spinal fluid of patients with meningitis in São Paulo, Brazil.

A total of 138 patients with the age of 4 months to 57 years were attended in different hospitals of São Paulo State with aseptic meningitis. A probable new agent was isolated from the cerebrospinal fluid of 35 of 53 specimens examined. Replication of the agent with similar characteristics was detected by CPE produced in the MDCK cell line. Virus-like particles measuring about 40 nm in diameter were observed by negative staining electron microscopy. No hemagglutinating activity was detected at pH 7.2 by using either human, guinea pig, chicken and at pH ranged 6.0-7.2 with goose red blood cells. The agent was not pathogenic to newborn or adult mice. Virus infectivity as measured by CPE was sensitive to chloroform and not inhibited by BuDR, suggesting that agent is an enveloped virus with RNA genome.

Visceral larva migrans: a seroepidemiological survey in five municipalities of São Paulo state, Brazil.

An enzyme-linked immunosorbent assay (ELISA), employing antigens from Toxocara canis larvae and the absortion of suspected sera with Ascaris lumbricoides extracts was used in a seroepidemiological study performed in five municipalities of São Paulo State, Brazil (São Paulo, Campinas, Santos, Marília and Presidente Prudente) in order to determine the frequency of antibodies to Toxocara. In 2,025 blood samples collected, 806 proceeded from male subjects and 1,219 from females; 483 samples were collected from subjects under 15 years of age and the remaining 1,542 from subjects aged 15 years or over. Among the 2,025 sera investigated, 3.60% had antibodies to Toxocara at significant levels. A moderate predominance of infection with Toxocara among male subjects (3.72%) was observed, although the difference was not statistically significant when this rate was compared with that for female (3.28%). Related to age, a higher frequency of positive results was detected among subjects under 15 years (6.41%) against the older group (2.53%). A trend of more elevated rates of infection was observed in municipalities with high demographic densities (São Paulo, Campinas and Santos). Nevertheless, such findings only appeared to be statistically significant in subjects younger than 15 years.