RESUMO
BACKGROUND: Respiratory virus-induced wheezing, such as that induced by respiratory syncytial virus (RSV) and human rhinovirus, is an important risk factor for recurrent wheezing and childhood asthma. However, no biomarkers for predicting recurrent wheezing have been identified. OBJECTIVE: We searched for predictors of recurrent wheezing using nasopharyngeal aspirates obtained from patients during the first wheezing episode who were hospitalized with an acute lower respiratory tract illness. METHODS: We enrolled 82 infants during the first wheezing episode (median age, 5.0 months) who were hospitalized for acute lower respiratory tract illness between August 2009 and June 2012 and followed these patients for 2.5 years. Nasopharyngeal aspirates and blood samples were obtained on the first day of hospitalization. Viral genomes were identified by using RT-PCR and sequencing. Levels of 33 cytokines, tryptase, IgE, anti-RSV IgE, and anti-RSV IgG were measured by using ELISAs or the Bio-Plex multiplex assay. Predictors of recurrent wheezing were examined by using a stepwise logistic regression model with backward elimination. RESULTS: Sixty percent of the patients experienced recurrent wheezing episodes. One or more viruses were detected in the nasopharynxes of 93% of the patients during the first wheezing episode. IFN-γ, IL-2, IL-9, MIP-1α, and MIP-1ß levels were significantly higher among patients with recurrent wheezing than among those without recurrent wheezing (P < .05 or .01). The stepwise model demonstrated that the MIP-1α level (odds ratio, 7.72; 95% CI, 1.50-39.77; P = .015) was the strongest independent predictor of the occurrence of recurrent wheezing. CONCLUSION: An increased MIP-1α level in nasopharyngeal aspirates from patients with acute respiratory symptoms during the first wheezing episode caused by viral infections might predict recurrent wheezing.
Assuntos
Quimiocina CCL3/metabolismo , Líquido Extracelular/metabolismo , Nasofaringe/metabolismo , Sons Respiratórios/diagnóstico , Anticorpos Antivirais/imunologia , Biomarcadores , Pré-Escolar , Citocinas/metabolismo , Feminino , Hospitalização , Humanos , Imunoglobulina E/imunologia , Imunoglobulina E/metabolismo , Imunoglobulina G , Lactente , Recém-Nascido , Masculino , Prognóstico , Recidiva , Sons Respiratórios/etiologia , Vírus Sinciciais Respiratórios/imunologia , TriptasesRESUMO
To improve detection of norovirus (NoVGI, NoVGII) and sapovirus (SaV), a simultaneous quantitative RT-PCR method was established. This triplex real-time PCR method was evaluated using a combination of optimized specific primers and probes. The performance of the developed PCR assay was equivalent to that of monoplex real-time PCR across a broad dynamic range of 10(2) -10(7) copies/assay using plasmid DNA standards. The limit of detection was 10(2) copies/assay. The quantitative value was comparable with that of monoplex real-time PCR of stool samples. Our triplex real-time PCR is useful for detection of NoV and SaV infections.
Assuntos
Reação em Cadeia da Polimerase Multiplex , Norovirus/genética , Reação em Cadeia da Polimerase em Tempo Real , Sapovirus/genética , Infecções por Caliciviridae/diagnóstico , Infecções por Caliciviridae/virologia , Humanos , Reprodutibilidade dos Testes , Sensibilidade e Especificidade , Carga Viral/métodosRESUMO
We isolated a novel influenza virus A(H1N2) strain from a pig on January 13, 2012, in Gunma Prefecture, Japan. Phylogenetic analysis showed that the strain was a novel type of double-reassortant virus derived from the swine influenza virus strains H1N1pdm09 and H1N2, which were prevalent in Gunma at that time.
Assuntos
Vírus da Influenza A Subtipo H1N1/genética , Vírus da Influenza A Subtipo H1N2/genética , Infecções por Orthomyxoviridae/veterinária , Vírus Reordenados/genética , Doenças dos Suínos/virologia , Animais , Genes Virais , Vírus da Influenza A Subtipo H1N1/isolamento & purificação , Vírus da Influenza A Subtipo H1N2/isolamento & purificação , Japão/epidemiologia , Dados de Sequência Molecular , Infecções por Orthomyxoviridae/epidemiologia , Infecções por Orthomyxoviridae/virologia , Filogenia , Vigilância em Saúde Pública , Vírus Reordenados/isolamento & purificação , Suínos , Doenças dos Suínos/epidemiologiaRESUMO
The present case provides direct evidence of human herpesvirus 6 reactivation in resected lymph node tissue in a patient with drug-induced hypersensitivity syndrome. This case clearly demonstrates that appropriate pathological evaluation of lymphadenopathy for drug-induced hypersensitivity syndrome, which mimics malignant lymphoma in clinical, radiological, and pathological findings, is required.
Assuntos
Hipersensibilidade a Drogas/complicações , Herpesvirus Humano 6/isolamento & purificação , Doenças Linfáticas/patologia , Doenças Linfáticas/virologia , Infecções por Roseolovirus/complicações , Infecções por Roseolovirus/diagnóstico , Ativação Viral , Feminino , Herpesvirus Humano 6/fisiologia , Humanos , Linfonodos/virologia , Pessoa de Meia-Idade , Infecções por Roseolovirus/virologiaRESUMO
Whole-genome sequencing of non-H(2)S-producing Salmonella enterica serovar Typhimurium and S. enterica serovar Infantis isolates from poultry meat revealed a nonsense mutation in the phsA thiosulfate reductase gene and carriage of a CMY-2 ß-lactamase. The lack of production of H(2)S might lead to the incorrect identification of S. enterica isolates carrying antimicrobial resistance genes.
Assuntos
Doenças das Aves Domésticas/microbiologia , Salmonelose Animal/microbiologia , Salmonella enterica/genética , Animais , Genoma Bacteriano , Sulfeto de Hidrogênio/metabolismo , Japão , Salmonella enterica/enzimologia , Salmonella enterica/isolamento & purificação , Salmonella enterica/metabolismo , Análise de Sequência de DNA , Sulfurtransferases/genética , beta-Lactamases/genéticaRESUMO
BACKGROUND: Respiratory syncytial virus (RSV) infection is associated with both the development and exacerbation of bronchial asthma. We examined eosinophil infiltration and the cytokine profiles of both airway and peripheral blood in antigen-sensitized mice infected with RSV to investigate the pathogenesis of exacerbations of asthma due to RSV infection. METHODS: Ovalbumin (OVA)-sensitized mice were challenged by OVA inhalation 3 times and then infected with RSV [10(5) TCID50 (50% of tissue culture infectious dose)/25 g body weight] or mock infection immediately after the last challenge. Animals from each group, namely, the control (PBS instead of OVA inhalation plus mock infection), RSV (PBS plus RSV), OVA (OVA plus mock) and OVA/RSV (OVA plus RSV) were analyzed. Analysis included evaluation of airway responsiveness to methacholine, pathological findings in the airway by hematoxylin and eosin (HE) and Luna staining, bronchoalveolar fluid (BALF) and peripheral leukocytes counts, and concentrations of multiple cytokines/chemokines in both BALF and serum. RESULTS: Airway responsiveness was significantly enhanced in the OVA and OVA/RSV groups compared with the control group. Levels of tissue and BALF eosinophils were higher in the OVA and OVA/RSV groups than in the RSV or control group. Significantly higher levels of macrophage inflammatory protein (MIP)-1α in BALF were observed in the OVA/RSV group compared with the 3 other groups. Production of serum IL-17 was also significantly elevated in the OVA/RSV group compared with the control or OVA group. CONCLUSIONS: These findings suggest that MIP-1α and IL-17 may play important roles in acute exacerbation of asthma induced by RSV in an animal model.
Assuntos
Asma/imunologia , Asma/virologia , Quimiocina CCL3/biossíntese , Interleucina-17/biossíntese , Infecções por Vírus Respiratório Sincicial/imunologia , Vírus Sinciciais Respiratórios/imunologia , Alérgenos/imunologia , Animais , Líquido da Lavagem Broncoalveolar/citologia , Líquido da Lavagem Broncoalveolar/imunologia , Linhagem Celular , Citocinas/biossíntese , Imunoglobulina E/sangue , Imunoglobulina E/imunologia , Pulmão/imunologia , Pulmão/patologia , Pulmão/virologia , Masculino , Camundongos , Ovalbumina/imunologia , Infecções por Vírus Respiratório Sincicial/metabolismoRESUMO
The cellular effects of eleven compounds including chalcone glycosides isolated from Brassica rapa L. 'hidabeni' and their synthetic derivatives were studied in rat pheochromocytoma PC12 cells. Of the compounds tested, 4'-O-ß-D-glucopyranosyl-3',4-dimethoxychalcone (A2) significantly increased the levels of the phosphorylated forms of extracellular signal-regulated kinases 1/2 (ERK 1/2), p38 mitogen-activated protein kinase (p38MAPK), and stress-activated protein kinases/Jun amino-terminal kinases (JNK/SAPK), but it did not affect Akt. Nerve growth factor (NGF), a well-known neurotrophic factor, increased the levels of phosphorylated ERK1/2, JNK/SAPK, and Akt but not p38MAPK, which may mediate marked neurite outgrowth. Signals evoked by A2 shared common characteristics with those induced by NGF; therefore, we evaluated the neuritogenic activity of A2 and found it induced only weak neurite outgrowth. However, this effect was enhanced by pre-treatment with a p38MAPK inhibitor, suggesting that the phosphorylation of p38MAPK down-regulated neurite outgrowth. From the results of this study, it was found that A2 in combination with a p38MAPK inhibitor can induce NGF-like effects. Hence, a combination of chalcone glycosides containing A2 and a p38MAPK inhibitor increases the likelihood that chalcone glycosides could be put to practical use in the form of drugs or alternative medicines to maintain neural health.
Assuntos
Chalconas/farmacologia , Neuritos/fisiologia , Proteínas Quinases p38 Ativadas por Mitógeno/antagonistas & inibidores , Animais , Chalconas/síntese química , Imidazóis/farmacologia , Sistema de Sinalização das MAP Quinases/efeitos dos fármacos , Sistema de Sinalização das MAP Quinases/fisiologia , Proteína Quinase 3 Ativada por Mitógeno/metabolismo , Neuritos/efeitos dos fármacos , Células PC12 , Piridinas/farmacologia , Ratos , Proteínas Quinases p38 Ativadas por Mitógeno/metabolismoRESUMO
To examine cytokine production in response to RSV infection, we assessed the levels of 29 cytokines released from RSV-infected human foetal lung fibroblasts. We also examined the relationships between the effects of fluticasone propionate and various signalling pathways in the cells. Twenty-four hours after infection (1MOI), RSV-infected cells released cytokines, for example proinflammatory cytokines (IL-1ß, IL-6 and TNF-α), anti-inflammatory (IL-1ra), Th1 (IFN-γ, IFN-λ1a, IL-2 and IL-12), Th2 (IL-4, IL-5, IL-10 and IL-13), granulopoiesis-inducing (G-CSF and GM-CSF), eosinophil recruitment-inducing (eotaxin and RANTES) and neutrophil recruitment-inducing cytokines (IL-8, IP-10, MCP-1 and MIP-1α). Aberrant release of most was significantly suppressed by fluticasone propionate. Twelve hours after RSV infection, increased phosphorylation of Akt, p38 MAPK, ERK1/2 and IκB-α was noted. Fluticasone propionate suppressed the phosphorylation of Akt, p38 MAPK, and ERK1/2, but not IκB-α, in virus-infected cells. TLR-4 expression was unchanged in control and RSV-infected cells, and TLR-3 and RIG-I expression was not detected. The results indicate that RSV infection induces aberrant production and release of certain cytokines through these signalling pathways in human lung fibroblasts. Overproduction and imbalance of these cytokines may be associated with the pathophysiology of RSV-induced excessive and allergic inflammation.
Assuntos
Androstadienos/farmacologia , Citocinas/metabolismo , Fibroblastos/metabolismo , Vírus Sinciciais Respiratórios/fisiologia , Transdução de Sinais , Anti-Inflamatórios/farmacologia , Linhagem Celular , Avaliação Pré-Clínica de Medicamentos , Fibroblastos/efeitos dos fármacos , Fibroblastos/imunologia , Fibroblastos/virologia , Fluticasona , Interações Hospedeiro-Patógeno , Humanos , Peptídeos e Proteínas de Sinalização Intracelular/metabolismo , Pulmão/citologia , Fosforilação , Processamento de Proteína Pós-Traducional , Receptores Toll-Like/metabolismoRESUMO
We studied the evolution of the G gene in the new genotype ON1 of RSV detected from patients with acute respiratory infection in Japan. Phylogenetic analyses and the evolutionary timescale were obtained by the Bayesian MCMC method. We also analyzed p-distance and positive selection sites. A new genotype ON1 emerged around 2001. The evolution rate was rapid (3.57 × 10(-3) substitutions/site per year). The p-distance was short and no positive selection site was found in the present strains. These results suggested that a new genotype ON1 of RSV-A emerged approximately10 years ago and spread to some countries with a high evolution rate.
Assuntos
Infecções por Vírus Respiratório Sincicial/virologia , Vírus Sincicial Respiratório Humano/genética , Vírus Sincicial Respiratório Humano/isolamento & purificação , Proteínas do Envelope Viral/genética , Sequência de Aminoácidos , Evolução Molecular , Genótipo , Humanos , Japão , Dados de Sequência Molecular , Filogenia , Vírus Sincicial Respiratório Humano/química , Vírus Sincicial Respiratório Humano/classificação , Alinhamento de Sequência , Proteínas do Envelope Viral/químicaRESUMO
Associations between the severity of respiratory signs and symptoms and the respiratory viruses identified in 214 Japanese children with acute respiratory illness (ARI) enrolled between January and December 2012 were studied. Respiratory rate, wheezing, cyanosis, and the use of accessory muscles were used as indices of respiratory severity and phylogenetic analysis of the viruses identified in these children was performed. Respiratory viruses such as respiratory syncytial virus (RSV), human rhinovirus (HRV), human parainfluenza virus (HPIV), and human metapneumovirus (HMPV) were prevalent, being detected in approximately 70% of the patients (151/214 patients). Co-detection of viruses occurred in about 9% of patients. RSV was identified more frequently in cases scored as moderate/severe than in those scored as mild (P < 0.05). Severity scores of patients with RSV were significantly higher than those of cases with HPIV. Moreover, severity scores in patients with mild disease and co-detections were higher than in those in whom only HPIV or adenovirus was detected. Phylogenetic analysis showed that many genotypes of HRV-A and -C with wide genetic divergence were associated with acute respiratory illness (ARI). On the other hand, only a limited number of genotypes of RSV were associated with ARI. HPIV and HMPV were associated with ARI at similar frequencies. These results suggest that different respiratory viruses with unique genetic characteristics can be found in patients with mild to severe ARI.
Assuntos
Infecções Respiratórias/patologia , Infecções Respiratórias/virologia , Índice de Gravidade de Doença , Viroses/patologia , Viroses/virologia , Criança , Pré-Escolar , Coinfecção/epidemiologia , Coinfecção/patologia , Coinfecção/virologia , Feminino , Genótipo , Humanos , Lactente , Japão/epidemiologia , Masculino , Dados de Sequência Molecular , Filogenia , Infecções Respiratórias/epidemiologia , Análise de Sequência de DNA , Viroses/epidemiologia , Vírus/classificação , Vírus/genética , Vírus/isolamento & purificaçãoRESUMO
OBJECTIVES: The objective of this study was to assess the need for and usefulness of training programs for Local Infectious Disease Surveillance Center (LIDSC) staff. METHODS: A structured questionnaire survey was conducted to assess the needs and usefulness of training programs. The subjects of the survey were participants of a workshop held after an annual conference for the LIDSC staff. Data on demographic information, the necessity of training programs for LIDSC staff, the themes and contents of the training program, self-assessment of knowledge on epidemiology and statistics were covered by the questionnaire. RESULTS: A total of 55 local government officials responded to the questionnaire (response rate: 100%). Among these, 95% of participants believed that the training program for the LIDSC staff was necessary. Basic statistical analysis (85%), descriptive epidemiology (65%), outline of epidemiology (60%), interpretation of surveillance data (65%), background and objectives of national infectious disease surveillance in Japan (60%), methods of field epidemiology (60%), and methods of analysis data (51%) were selected by over half of the respondents as suitable themes for training programs. A total of 34 LIDSC staff answered the self-assessment question on knowledge of epidemiology. A majority of respondents selected "a little" or "none" for all questions about knowledge. Only a few respondents had received education in epidemiology. CONCLUSION: The results of this study indicate that LIDSC staff have basic demands for fundamental and specialized education to improve their work. Considering the current situation regarding the capacity of LIDSC staff, these training programs should be started immediately.
Assuntos
Controle de Doenças Transmissíveis , Educação Continuada , Monitoramento Epidemiológico , Humanos , JapãoRESUMO
OBJECTIVE: Granulocyte-colony stimulating factor (G-CSF) and vascular endothelial growth factor (VEGF) are thought to be associated with the pathophysiology of perinatal asphyxia. To clarify any such association, we analyzed the serum levels in neonates with perinatal asphyxia treated with head cooling. STUDY DESIGN: Temporal alterations of serum G-CSF and VEGF levels were measured within 24h of birth in five neonatal cases of severe asphyxia treated with head cooling, five neonatal cases without head cooling, and four healthy neonatal cases. RESULTS: G-CSF in sera markedly increased and sustained in severely asphyxiated neonates treated with head cooling, while VEGF decreased and remained low. CONCLUSION: G-CSF and VEGF levels in sera might be associated with an early phase of brain protection after birth in severe asphyxia treated with head cooling.
Assuntos
Asfixia Neonatal/sangue , Crioterapia , Fator Estimulador de Colônias de Granulócitos/sangue , Fator A de Crescimento do Endotélio Vascular/sangue , Asfixia Neonatal/metabolismo , Asfixia Neonatal/terapia , Encéfalo/metabolismo , Temperatura Baixa , Citocinas/sangue , Cabeça , Humanos , Recém-NascidoRESUMO
BACKGROUND: Respiratory syncytial (RS) virus infection is an important exacerbating factor in acute bronchial asthma. However, the precise mechanisms responsible for viral infection-induced exacerbations of asthma are uncertain. To elucidate the role of eosinophilic inflammation in the pathogenesis of virus-induced asthma, we investigated the effects of eosinophil granule proteins on bronchial epithelial cell infected with RS virus. METHODS: Morphological changes and cytopathic effects in human type II pulmonary alveolar epithelial cells (A549) infected with RS virus and/or eosinophil granule proteins such as major basic protein (MBP), eosinophil peroxidase (EPO), eosinophil cationic protein (ECP) and eosinophil-derived neurotoxin (EDN) were observed by microscopy. Apoptosis/necrosis was evaluated by trypan blue exclusion test. We also measured 8 types of phosphorylated proteins in MBP-treated A549 cells infected with RS virus. RESULTS: Although RS virus alone did not affect the cytopathic effects of A549 cells, high concentrations of MBP or a combination of 4 granule proteins resulted in cytopathic effects. MBP or EPO, but not ECP or EDN, induced cytotoxicity and necrosis of the infected A549 cells. Furthermore, MBP induced the phosphorylation of the extracellular signal-regulated kinase (ERK) 1/2, p38 mitogen-activated protein kinase (MAPK), Jun-N-terminal protein kinase (JNK), and signal transducer and activator of transcription (STAT)3 in the infected cells. CONCLUSIONS: These results suggest that eosinophil granule proteins, specifically MBP, damage bronchial epithelial cells infected with RS virus and that the MAPK family are involved in these responses, indicating that eosinophilic inflammation might be associated with the pathophysiology of RS virus-induced acute exacerbations of asthma.
Assuntos
Brônquios/imunologia , Proteínas Granulares de Eosinófilos/imunologia , Células Epiteliais/imunologia , Infecções por Vírus Respiratório Sincicial/imunologia , Brônquios/citologia , Linhagem Celular Tumoral , Sobrevivência Celular , Humanos , Proteínas Quinases Ativadas por Mitógeno/imunologia , Fator de Transcrição STAT3/imunologiaRESUMO
OBJECTIVES: Leptospirosis is a zoonotic infection; water and soil are contaminated with Leptospira through the urine of rodents carrying the agent in the kidneys, and then the infection is established percutaneously or orally. In November 2009, the first notification of leptospirosis was submitted in Gunma Prefecture. It is important to clarify the current status of the leptospirosis epidemic in the prefecture and to enlighten residents and healthcare professionals. Hence, we examined the infection status of leptospirosis in residents living near the residence of the patient in this study. SUBJECTS: SUBJECTS were residents of Isesaki City, Gunma Prefecture, aged 40-64 years and who had undergone a specific health examination (mass health examination) for insured persons under Japan's National Health Insurance on April 23 and 26, 2010. SAMPLE SIZE: We randomly selected 100 subjects from residents who underwent the examination, and antibody titer against leptospirosis was measured. Testing method: Measurement of the antibody titer was commissioned to the National Institute of Infectious Diseases. Microscopic agglutination tests using 6 serotypes of living Leptospira that are widely distributed to the main islands of Japan were performed. RESULTS: One hundred eighty subjects underwent the examination on the 2 days, and 174 of them consented to participate in the study. Serum antibody titer was measured in 100 patients, and the results were all negative. CONCLUSION: Leptospirosis is a treatable disease when it is diagnosed; therefore, early diagnosis and treatment is important. Mild cases may be missed. Awareness of leptospirosis is low among healthcare professionals in the prefecture. Although it is a rarely infectious disease, it is important to make leptospirosis known to medical associations in the prefecture and to conduct training workshops and similar activities.
Assuntos
Leptospirose/epidemiologia , Adulto , Anticorpos Antibacterianos/sangue , Feminino , Humanos , Japão/epidemiologia , Leptospirose/imunologia , Masculino , Pessoa de Meia-Idade , Estudos SoroepidemiológicosRESUMO
Synthetic organic selenium compounds, such as ebselen, may show glutathione peroxidase-like antioxidant activity and have a neurotrophic effect. We synthesized 1,3-selenazolidin-4-ones, new types of synthetic organic selenium compounds (five-member ring compounds), to study their possible applications as antioxidants or neurotrophic-like molecules. Their superoxide radical scavenging effects were assessed using the quantitative, highly sensitive method of real-time kinetic chemiluminescence. At 166µM, the O(2)(-) scavenging activity of 1,3-selenazolidin-4-ones ranged from 0 to 66.2%. 2-[3-(4-Methoxyphenyl)-4-oxo-1,3-selenazolidin-2-ylidene]malononitrile (compound b) showed the strongest superoxide anion-scavenging activity among the 6 kinds of 2-methylene-1,3-selenazolidin-4-ones examined. Compound b had a 50% inhibitory concentration (IC(50)) at 92.4µM and acted as an effective and potentially useful O(2)(-) scavenger in vitro. The effect of compound b on rat pheochromocytome cell line PC12 cells was compared with that of ebselen or nerve growth factor (NGF) by use of the MTT [3-(4, 5-dimethyl-2-thiazolyl)-2,5-diphenyl-2H-tetrazolium bromide] assay. When ebselen was added at 100µM or more, toxicity toward PC12 cells was evident. On the contrary, compound b suppressed serum deprivation-induced apoptosis in PC12 cells more effectively at a concentration of 100µM. The activity of compound b to phosphorylate mitogen-activated protein kinase/extracellular signal-regulated protein kinase (ERK) 1/2 (MAP kinase) in PC12 cells was higher than that of ebselen, and the former at 100µM induced the phosphorylation of MAP kinase to a degree similar to that induced by NGF. From these results, we conclude that this superoxide anion-scavenger, compound b, suppressed serum deprivation-induced apoptosis by promoting the phosphorylation of MAP kinase.
Assuntos
Apoptose/efeitos dos fármacos , Azóis/farmacologia , Sequestradores de Radicais Livres/farmacologia , Compostos Organosselênicos/farmacologia , Superóxidos/metabolismo , Animais , Azóis/síntese química , Azóis/química , Relação Dose-Resposta a Droga , Sequestradores de Radicais Livres/síntese química , Sequestradores de Radicais Livres/química , Concentração Inibidora 50 , Isoindóis , Medições Luminescentes , Proteína Quinase 1 Ativada por Mitógeno/metabolismo , Proteína Quinase 3 Ativada por Mitógeno/metabolismo , Fator de Crescimento Neural/farmacologia , Compostos Organosselênicos/síntese química , Compostos Organosselênicos/química , Células PC12 , Fosforilação/efeitos dos fármacos , Ratos , Relação Estrutura-AtividadeRESUMO
BACKGROUND: Human parainfluenza virus type 1 (HPIV1) causes various acute respiratory infections (ARI). Hemagglutinin-neuraminidase (HN) glycoprotein of HPIV1 is a major antigen. However, the molecular epidemiology and genetic characteristics of such ARI are not exactly known. Recent studies suggested that a phylogenetic analysis tool, namely the maximum likelihood (ML) method, may be applied to estimate the evolutionary time scale of various viruses. Thus, we conducted detailed genetic analyses including homology analysis, phylogenetic analysis (using both the neighbor joining (NJ) and ML methods), and analysis of the pairwise distances of HN gene in HPIV1 isolated from patients with ARI in Yamagata prefecture, Japan. RESULTS: A few substitutions of nucleotides in the second binding site of HN gene were observed among the present isolates. The strains were classified into two major clusters in the phylogenetic tree by the NJ method. Another phylogenetic tree constructed by the ML method showed that the strains diversified in the late 1980s. No positively selected sites were found in the present strains. Moreover, the pairwise distance among the present isolates was relatively short. CONCLUSIONS: The evolution of HN gene in the present HPIV1 isolates was relatively slow. The ML method may be a useful phylogenetic method to estimate the evolutionary time scale of HPIV and other viruses.
Assuntos
Proteína HN/genética , Epidemiologia Molecular , Vírus da Parainfluenza 1 Humana/genética , Filogenia , Infecções Respiratórias/epidemiologia , Infecções por Respirovirus/epidemiologia , Doença Aguda , Adolescente , Adulto , Animais , Linhagem Celular , Criança , Pré-Escolar , Proteína HN/química , Humanos , Lactente , Recém-Nascido , Japão/epidemiologia , Funções Verossimilhança , Dados de Sequência Molecular , Vírus da Parainfluenza 1 Humana/isolamento & purificação , Infecções Respiratórias/virologia , Infecções por Respirovirus/virologia , Análise de Sequência de DNA , Adulto JovemRESUMO
Because little information is available on eosinophil activation and cytokine response in virus-induced wheezing, we attempted to detect respiratory viruses and measure eosinophil cationic protein (ECP), and 27 types of cytokines/chemokines in both serum and nasal secretions from children with wheezing. This study was an observational, case-control investigation of 267 subjects, who were visited and/or hospitalized with acute respiratory symptoms (with wheezing: men, 115; women, 59; mean/median age, 3.6/3.0 years) or who were visited for regular physical examination and treatment (non-symptomatic wheezing: men, 48; women, 31; mean/median, 5.0/4.7 years), and 14 control subjects (controls: men, 9; women, 5; mean/median, 3.6/3.7 years). We detected viruses in nasal secretions from 174 patients with acute exacerbations of wheezing using antigen detection kits or reverse transcription-polymerase chain reaction, followed by direct DNA sequencing analysis. We measured peripheral eosinophil counts, and serum concentrations of ECP and 27 cytokines/chemokines using a multiplex bead-based assay in patients with wheezing or non-symptomatic wheezing. We also examined nasal ECP and 27 cytokines/chemokines in patients with wheezing. Of 174 samples from wheezing exacerbations, rhinovirus was detected in 59; respiratory syncytial (RS) virus in 44; enterovirus in 17; other viruses in 19; and no viruses in 35. Serum concentrations of ECP, IL-5, IL-6, IL-1ra, and IP-10 were significantly elevated in rhinovirus-induced wheezing compared with non-symptomatic wheezing. Similarly, serum ECP, IL-5, and IP-10 were significantly higher in rhinovirus-induced wheezing than in controls. On the other hand, IL-1ra and IP-10, but not ECP and IL-5 were significantly higher in RS virus-induced wheezing than in controls. Furthermore, only IL-5 was significantly elevated in the rhinovirus group compared with the RS virus group in both serum and nasal secretions. Different cytokine profile and eosinophil activation might be involved in rhinovirus- and RS virus-induced acute exacerbation of childhood wheezing.
Assuntos
Resfriado Comum/complicações , Citocinas/metabolismo , Eosinófilos/imunologia , Sons Respiratórios/etiologia , Infecções por Vírus Respiratório Sincicial/complicações , Vírus Sincicial Respiratório Humano/isolamento & purificação , Rhinovirus/isolamento & purificação , Doença Aguda , Estudos de Casos e Controles , Pré-Escolar , Resfriado Comum/diagnóstico , Resfriado Comum/imunologia , Resfriado Comum/virologia , Proteína Catiônica de Eosinófilo/sangue , Proteína Catiônica de Eosinófilo/metabolismo , Feminino , Humanos , Masculino , Sons Respiratórios/fisiopatologia , Infecções por Vírus Respiratório Sincicial/diagnóstico , Infecções por Vírus Respiratório Sincicial/imunologia , Infecções por Vírus Respiratório Sincicial/virologia , Vírus Sincicial Respiratório Humano/genética , Vírus Sincicial Respiratório Humano/imunologia , Rhinovirus/genética , Rhinovirus/imunologiaRESUMO
RSV (respiratory syncytial virus)-induced pneumonia and bronchiolitis may be associated with hyperresponsive conditions, including asthma. Eosinophilic proteins such as MBP (major basic protein) may also be associated with the pathophysiology of asthma. To elucidate the roles of RSV infection and MBP in the pathogenesis of pneumonia with hyperresponsiveness, we investigated the effects of RSV infection and MBP on A549 (alveolar epithelial) cells. CPE (cytopathic effects) in A549 cells were observed by microscopy. Apoptosis and cell death was evaluated by flow cytometric analysis and modified MTT [3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide] assay. We also measured 15 types of cytokines and chemokines in A549 cell supernatants. Although RSV alone did not affect the CPE of A549, high concentrations of MBP resulted in cell death within 24 h. Combinations of RSV and MBP synergistically induced cell death. In A549 cells infected with RSV alone, the release of GM-CSF (granulocyte-macrophage colony-stimulating factor) was significantly enhanced compared with control cells (no infection). In the cells treated with MBP alone, the production of IL (interleukin)-2, 4, 5, 7, 10, 12, 13, 17, IFN (interferon)-γ, GM-CSF, G-CSF (granulocyte colony-stimulating factor) and MIP (macrophage inflammatory protein)-1ß was significantly increased compared with control cells. Notably, the levels of GM-CSF and IL-17 in RSV/MBP-treated cells were significantly higher than those treated with MBP alone. These results suggest that MBP synergistically enhanced the release of various cytokines/chemokines and the cell death of RSV-infected A549 cells, indicating that MBP may be closely associated with the pathophysiology of allergic reactions in bronchiolitis/pneumonia due to RSV.
Assuntos
Proteína Básica Maior de Eosinófilos/imunologia , Pneumonia/imunologia , Pneumonia/virologia , Alvéolos Pulmonares/patologia , Alvéolos Pulmonares/virologia , Infecções por Vírus Respiratório Sincicial/complicações , Linhagem Celular , Sobrevivência Celular , Quimiocinas/imunologia , Eosinófilos/imunologia , Células Epiteliais/patologia , Células Epiteliais/virologia , Humanos , Pneumonia/etiologia , Pneumonia/patologia , Alvéolos Pulmonares/citologiaRESUMO
We newly synthesized organic selenium compounds (5-membered ring compounds) including 2-selenoxo-1,3-thiazolidin-4-ones (compounds A) and 3-alkoxy-4,5-dihydro-5-selenoxo-1H-1,2,4-triazole-1-carboxylates (compounds B). To address whether these compounds show antioxidative effects, we also examined their superoxide radical (O(2) (-))-scavenging effects. Moreover, we examined the effects of compound Aa on the activation of mitogen-activated protein kinase/extracellular signal-regulated protein kinases (MAPK/ERK1/2) and suppression of hydrogen peroxide-induced cytotoxicity in rat pheochromocytoma cells (PC12 cells). We evaluated the O(2) (-)-scavenging activities of the compounds by a chemiluminescence method, and activation of ERK1/2 in PC12 cells was evaluated by Western blot analysis. At 166 µmol/L, the O(2) (-)-scavenging activities were markedly different among compounds A and B. 3-(2,6-Dimethylphenyl)-2-selenoxo-1,3-thiazolidin-4-one (compound Aa) exhibited the strongest superoxide anion-scavenging activity among compounds A and B. The concentration necessary for 50% inhibition of the activity (IC(50)) of compound Aa was 25.9 µmol/L. Compound Aa activated ERK1/2 of the PC12 cell, as did ebselen, and suppressed hydrogen peroxide-induced cytotoxicity more potently than ebselen. In addition, the toxicity of compound Aa was less than that of ebselen. From these results, it is assumed that compound Aa is a candidate drug to prevent oxidative stress-induced cell death.
Assuntos
Antioxidantes/farmacologia , Peróxido de Hidrogênio/toxicidade , Proteínas Quinases Ativadas por Mitógeno/metabolismo , Compostos Organosselênicos/farmacologia , Animais , Antioxidantes/síntese química , Proliferação de Células/efeitos dos fármacos , Sobrevivência Celular/efeitos dos fármacos , Compostos Organosselênicos/síntese química , Células PC12 , Ratos , Superóxidos/metabolismoRESUMO
Koplik spots are considered a disease-specific sign for measles, although comprehensive virological studies have not been conducted to date. In Japan, a national survey of 3023 measles and measles-suspected cases was conducted between 2009 and 2014 using polymerase chain reaction (PCR) or reverse transcription PCR (RT-PCR) to detect various rash/fever-associated viruses. Koplik spots were observed in 717 of 3023 cases (23.7%). Among these, the measles virus was detected in 202 cases (28.2%), while the rubella virus was detected in 125 cases (17.4%). Other viruses were detected in 51 cases having the spots (7.1%). In some of the cases with spots, two or three viruses, such as the rubella virus, parvovirus, and human herpesvirus type 6 were also detected. The sensitivity and specificity of Koplik spots as a diagnostic marker for measles were 48 and 80%, respectively. The results suggested that Koplik spots might appear not only in measles but also in other viral infections, such as rubella, as a clinical sign.