RESUMO
In Japan, a national project of longitudinal health care and epidemiological research (NEWS) was developed in 2014 to analyse the effects of radiation on human health for workers who responded to the Fukushima Dai-ichi nuclear emergency in 2011. In 2018, peripheral blood for chromosome translocation analysis was collected from 62 workers. Retrospective dose assessment was performed with fluorescence in situ hybridisation translocation (FISH-Tr) assay. The range of estimated doses by FISH-Tr assay was 0-635 mGy, in which 22 workers had estimated doses of more than 189 mGy. Biological dose estimates were five times higher in workers with physically measured total exposure recordings above 70 mGy. It is likely that smoking and medical exposure caused the discrepancy between estimated biological and physical total exposure doses. Thus, there is a possibility that retrospective biodosimetry assessment might over-estimate occupational exposures to workers exposed to chronic radiation during nuclear emergency work.
Assuntos
Bioensaio , Translocação Genética , Humanos , Estudos Retrospectivos , Instalações de Saúde , JapãoRESUMO
We developed a new quantification method for the measles virus (MeV) nucleoprotein (N) gene using real-time reverse transcriptase PCR. This method allowed us to quantify 10(1)-10(7) copies per reaction (corresponding to 5x10(-1)-5x10(5) copies microl(-1)) of the MeV N gene. We also quantified the MeV N gene from the throat swabs of 22 patients with measles as well as the MeV genotypes A, D3, D5, D9 and H1 in viral suspensions derived from MeV-infected cells. As a result, 3.9x10(3)-5.2x10(6) copies ml(-1) and 7.4x10(7)-2.0x10(8) copies ml(-1) of the MeV genomes (N gene) were detected in the throat swabs and viral suspensions, respectively. No other viruses (enteroviruses, respiratory syncytial virus, human metapneumovirus or mumps virus) were detected in the assay. The results suggest that this method is applicable to the detection and quantification of some genotypes of MeV.
Assuntos
Vírus do Sarampo/genética , Sarampo/virologia , Proteínas do Nucleocapsídeo/genética , Reação em Cadeia da Polimerase Via Transcriptase Reversa/métodos , Primers do DNA , Amplificação de Genes , Genes Virais/genética , Genótipo , Humanos , Vírus do Sarampo/isolamento & purificação , Dados de Sequência Molecular , Faringe/virologia , Plasmídeos , RNA Viral/genética , RNA Viral/isolamento & purificaçãoRESUMO
This report describes multiple viruses in stool specimens from oyster-associated gastroenteritis. Eleven outbreaks of oyster-associated gastroenteritis were examined for enteric viruses between January 2002 and March 2006 in Japan. Multiple norovirus genotypes were detected in all outbreaks; moreover, kobuvirus, sapovirus, and astrovirus were also detected in 6, 3, and 1 of the 11 outbreaks, respectively. Notably, multiple sapovirus genogroups were detected in the stool specimens from subjects in two oyster-associated gastroenteritis outbreaks.
Assuntos
Surtos de Doenças , Fezes/virologia , Gastroenterite/epidemiologia , Norovirus/isolamento & purificação , Ostreidae/virologia , Sapovirus/isolamento & purificação , Frutos do Mar/virologia , Animais , Infecções por Caliciviridae/epidemiologia , Infecções por Caliciviridae/virologia , Contaminação de Alimentos , Gastroenterite/genética , Gastroenterite/virologia , Genótipo , Humanos , Japão/epidemiologia , Norovirus/classificação , Ostreidae/genética , Filogenia , RNA Viral/genética , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Sapovirus/classificaçãoRESUMO
Loss of heterozygosity (LOH) plays an important role in leukemogenesis via inactivation of tumor suppressor genes. Recent studies have demonstrated that mouse thymic lymphomas (TL), a suitable model for the mechanistic study of human acute lymphoblastic leukemia, show frequent LOH on chromosomes 4 (p15/p16), 11 (Ikaros), 12 (Bcl11b), 19 (Pten), and X. To date, however, little data are available regarding the mechanism of LOH. In this study, we re-evaluated chromosomal abnormality and loss of heterozygosity in 26 TL-induced by x-rays in C57BL/6 (B6), C3H, and F1 (B6 x C3H) mice, using the quinacrine-Hoechst double-staining method and chromosome painting technique. Chromosomally abnormal cells were present in 25 TL examined (25/26, 96%). The most frequent abnormality was trisomy or partial trisomy of chromosome 15 (16/26, 62%), which is consistent with previous studies. Structural abnormalities of chromosome 11 with interstitial deletion of proximal region and chromosome 12 with translocation with deletion of distal region were newly identified in 7 (27%) and 12 (46%) cases, respectively. These results indicate that the distinct mechanism contributes to the LOH of each tumor suppressor gene on different chromosomal locations.
Assuntos
Aberrações Cromossômicas , Cromossomos de Mamíferos , Perda de Heterozigosidade/genética , Linfoma/genética , Neoplasias do Timo/genética , Animais , Coloração Cromossômica , Hibridização in Situ Fluorescente , Cariotipagem , Camundongos , Camundongos Endogâmicos C57BL , Camundongos Endogâmicos , Trissomia , Raios XRESUMO
A total of 225 oysters from China and Japan were collected during October 2005 to September 2006 and were then tested for the presence of norovirus by RT-nested PCR. The detection rate of norovirus was different between China and Japan, accounting for 14.6% (19 of 130) and 25.3% (24 of 95), respectively. In China, norovirus in oyster was detected continuously from July to February with the highest prevalence in August, October and November (each of 21%, 4 of 19). On the other hand, norovirus in Japan was found year-round with highest prevalence in March and October (each of 20.8%, 5 of 24). Norovirus strains detected were subjected to further characterization by sequence analysis. It was found that the norovirus strains belonged to only two distinct genotypes, the GII/3 (known as the Mexico virus cluster) and the GII/4 (known as the Lordsdale virus cluster). In China, the norovirus GII/4 was the most predominant, accounting for 78.9% (15 of 19). In contrast, it was interesting that both the norovirus GII/4 and the norovirus GII/3 were co-predominant with a prevalence of 50% (12 of 24) in Japan. Another interesting feature of the study was that the norovirus GII/4 strains in oysters from both countries were grouped into two distinct variant clusters known as the Farmington Hills variant and the Hunter variant. More than 102 copies of norovirus were detected in 41 of 43 oysters. This study provided additional evidence of the presence of norovirus in oysters and is also the first report to demonstrate the existence of norovirus variants in oysters.
Assuntos
Norovirus/genética , Ostreidae/virologia , Animais , China , Análise por Conglomerados , Variação Genética , Genótipo , Japão , Norovirus/classificação , Norovirus/isolamento & purificação , Ostreidae/química , Filogenia , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Estações do AnoRESUMO
In May 2004, 65 people from 18 groups of visitors to guesthouse (a traditional Japanese guesthouse) in the Nagano Prefecture, Japan developed acute gastroenteritis. Although these cases originally attributed to food poisoning, based on epidemiological and dietary surveys, there was nothing that is associated as a cause food. The same wall water was used throughout the guesthouse except in the kitchen, so testing was conducted on this water. Lordsdale variant strain of Norovirus was detected from both of the well water and the feces of patients and staff. The well supplying to the guesthouse was only 10 meters deep and fecal coliform group was also detected in the well water from the guesthouse. This suggested that the water source was contaminated by human feces.
Assuntos
Surtos de Doenças , Contaminação de Alimentos , Gastroenterite/virologia , Norovirus/isolamento & purificação , Poluição da Água , Abastecimento de Água , Doença Aguda , Humanos , JapãoRESUMO
In order to estimate biological doses after low-dose ionizing radiation exposure, fluorescence in situ hybridization (FISH) using three differentially colored chromosome painting probes was employed to detect exchange-type chromosome aberrations. A reference dose response curve was constructed using blood samples from a female donor whose lymphocytes consistently exhibited a low frequency of cells at the second mitosis under routine culture conditions. Aberration yields were studied for a total of about 155 thousand metaphases obtained from seven dose-points of gamma irradiations (0, 50, 100, 150, 200, 250 and 300mGy). In situ hybridization was performed using commercially available painting probes for chromosomes 1, 2 and 4. With the aid of an automated image-capturing method, exchange-type aberrations involving painted chromosomes were detected with considerable accuracy and speed. The results on the exchange-type aberrations (dicentrics plus translocations) at the seven dose-points showed a good fit to the linear-quadratic model (y=0.0023+0.0015x+0.0819x(2), P=0.83). A blind test proved the reproducibility of the reference dose-response relationship. In the control experiments using blood samples from another donor, the estimated doses calculated on the basis of the present reference curve were proved to be in good agreement with the actual physical doses applied. The present dose-response curve may serve as a means to assess the individual differences in cytogenetical radio-sensitivities.
Assuntos
Aberrações Cromossômicas/efeitos da radiação , Raios gama , Linfócitos/efeitos da radiação , Adulto , Relação Dose-Resposta à Radiação , Feminino , Humanos , Hibridização in Situ Fluorescente , Metáfase , Reprodutibilidade dos Testes , Translocação GenéticaRESUMO
The dicentric chromosome assay (DCA) has been regarded as the gold standard of radiation biodosimetry. The assay, however, requires a 2-d peripheral blood lymphocyte culture before starting metaphase chromosome analyses to estimate biological doses. Other biological assays also have drawbacks with respect to the time needed to obtain dose estimates for rapid decision on the correct line of medical treatment. Therefore, alternative technologies that suit requirements for triage biodosimetry are needed. Radiation-induced DNA double strand breaks in G0 lymphocytes can be detected as interphase chromosome aberrations by the cell fusion-mediated premature chromosome condensation (PCC) method. The method, in combination with fluorescence in situ hybridization (FISH) techniques, has been proposed in early studies as a powerful tool for obtaining biological dose estimates without 2-d lymphocyte culture procedures. The present work assesses the applicability of FISH-based PCC techniques using pan-centromeric and telomeric peptide nucleic acid (PNA) probes in triage mode biodosimetry and demonstrates that an improved rapid procedure of the prematurely condensed dicentric chromosome (PCDC) assay has the potential for evaluating exposed radiation doses in as short as 6 h after the collection of peripheral blood specimens.
Assuntos
Aberrações Cromossômicas/efeitos da radiação , Hibridização in Situ Fluorescente/métodos , Radiometria/métodos , Triagem/métodos , Adulto , Feminino , Humanos , Fatores de TempoRESUMO
A party of 57 people dined together in a restaurant in Hamamatsu City on December 11, 2001. The next day, 22 of them developed symptoms of acute gastroenteritis, such as diarrhea, vomiting, and fever. Examination of 4 fecal specimens from these patients by ELISA for Norovirus (Norwalk-like virus, NV) detected both genogroup I (GI) and genogroup II (GII) NV in all the 4 specimens. In addition, RT-PCR and real-time PCR methods for NV detected the NV gene. Approximately one month after the outbreak of the food poisoning (acute gastroenteritis) by NV, 4 individuals in the same party developed type A hepatitis. Both RT-PCR and real-time PCR methods for hepatitis A virus (HAV) detected the HAV gene in their fecal specimens. The party of these patients ate purple Washington clam (Saxidomus purpuratus, imported from China) steamed with red pepper. Since this food appeared to have caused the viral infections, the one with the same lot number was subjected to viral examinations, which successfully detected the NV GI, NV GII, and HAV genes. These results led to the conclusion that the clam contaminated with NV and HAV had caused the food poisoning. The DNA sequences of the NV detected in the patients and the clam had 74 to 99% homology, indicating strains of various genotypes. All the strains of HAV that were derived from the patients and the clam were genotype 1A, and these sequences had over 95% homology, but were not completely identical. This outbreak led to the demonstration of imported fishery products as a cause of type A hepatitis, and indicated the need for guiding and enlightening people on the importance of adequate cooking of bivalves.
Assuntos
Bivalves/virologia , Infecções por Caliciviridae/epidemiologia , Surtos de Doenças , Doenças Transmitidas por Alimentos/epidemiologia , Hepatite A/epidemiologia , Animais , Doenças Transmitidas por Alimentos/virologia , Gastroenterite/epidemiologia , Vírus da Hepatite A/isolamento & purificação , Humanos , Norovirus/isolamento & purificação , Frutos do Mar/virologiaRESUMO
PURPOSE: JF1/Ms mice, an inbred strain derived from Japanese wild mice, carry a germline hypomorphic mutation in the endothelin receptor type B gene (Ednrb). We observed that the JF1/Ms mice develop various spontaneous tumors at a high incidence late in life. The aim of this study was to elucidate the mechanism responsible for spontaneous tumors in these mice. Possible relevance of milk-borne mammary tumor virus and gene alterations in Ednrb to tumorigenesis was explored. METHODS: Expression and methylation status of Ednrb were quantitatively analyzed in normal and cancer tissues of mammary gland, liver, submandibular gland as well as in a cultured cell line, MW1, established from a submandibular gland adenocarcinoma. The biological effects of EDNRB were examined in the MW1 cells transfected with wild-type Ednrb. RESULTS: Transcripts of Ednrb were barely detectable, and the promoter region of Ednrb was hypermethylated in tumor tissues and the MW1 cells. In contrast, normal counterpart tissues showed positive expression of Ednrb transcripts and had unmethylated promoter regions. Treatment of the MW1 cells with 5-Aza-dC restored transcription of Ednrb to normal levels. Transfection of the MW1 cells with Ednrb1 (MW1-Ednrb1) resulted in lower growth rates and morphological changes compared with the mock-transfected MW1 cells (MW1-mock1). Furthermore, the MW1-Ednrb1 cells transplanted in syngeneic mice showed a lower proliferation rate than the MW1-mock1 cells. CONCLUSIONS: Germline mutation and subsequent promoter methylation of Ednrb may be relevant to cancer susceptibility in the JF1/Ms mice. These data indicate that Ednrb acts as a tumor suppressor, as reported in human prostate, bladder, and clear cell renal carcinomas.
Assuntos
Metilação de DNA , Mutação em Linhagem Germinativa , Camundongos Endogâmicos/genética , Neoplasias Experimentais/genética , Receptor de Endotelina B/genética , Animais , Carcinogênese/genética , Linhagem Celular Tumoral , Feminino , Genes Supressores de Tumor , Neoplasias Mamárias Experimentais/genética , Neoplasias Mamárias Experimentais/patologia , Neoplasias Mamárias Experimentais/virologia , Vírus do Tumor Mamário do Camundongo/isolamento & purificação , Camundongos , Camundongos Endogâmicos C57BL , Neoplasias Experimentais/patologia , Neoplasias Experimentais/virologia , Regiões Promotoras GenéticasRESUMO
The biological dose of nuclear workers engaged in emergency response tasks at Tokyo Electric Power Company (TEPCO) Fukushima Daiichi Nuclear Power Station was estimated in the present study. As the national core center for radiation emergency medical preparedness in Japan, the National Institute of Radiological Sciences (NIRS) received all individuals who were suspected of being overexposed to acute radiation. In the course of health examinations at NIRS, biological dosimetry was performed by the dicentric chromosome assay (DCA). Twelve individuals were examined from 21 March-1 July 2011. The results indicated that the estimated exposure doses for all individuals were lower than 300 mGy, with the mean value of about 101 mGy. These results by DCA were in accordance with those obtained by physical dosimetry based on personal dosimeter recording assessment. The results corroborate the fact that no acute radiation syndrome was observed among the workers examined.
Assuntos
Acidente Nuclear de Fukushima , Centrais Nucleares , Exposição Ocupacional/análise , Radiometria/métodos , Adulto , Cromossomos Humanos/efeitos da radiação , Humanos , Japão , Masculino , Doses de Radiação , Adulto JovemAssuntos
Infecções por Caliciviridae/transmissão , Infecções por Caliciviridae/virologia , Doenças Transmitidas por Alimentos/virologia , Gastroenterite/virologia , Norovirus , Animais , Infecções por Caliciviridae/epidemiologia , Infecções por Caliciviridae/prevenção & controle , Surtos de Doenças , Fezes/virologia , Doenças Transmitidas por Alimentos/epidemiologia , Doenças Transmitidas por Alimentos/prevenção & controle , Gastroenterite/epidemiologia , Gastroenterite/prevenção & controle , Conteúdo Gastrointestinal/virologia , Humanos , Japão/epidemiologia , Norovirus/genética , Norovirus/isolamento & purificação , Norovirus/patogenicidade , Ostreidae/virologiaRESUMO
We performed phylogenetic and cluster analysis of human rhinovirus species A (HRV-A) isolated from 76 children with acute respiratory infection in Yamagata prefecture, Japan during the period 2003-2007. Phylogenetic trees based on the nucleotide and amino acid sequences of the VP4/VP2 coding region showed that the present strains could be classified into 11 and 8 clusters, respectively. The homology among the present strains ranged from 66.6% to 100% at the nucleotide level and 84.7% to 100% at the amino acid level. The interspecies distance (mean+/-standard deviation) was calculated to be 0.235+/-0.048 at the nucleotide level and 0.076+/-0.033 at the amino acid level. In addition, the phylogenetic trees created based on the nucleotide and amino acid sequences showed that HRV-A strains belonging to some clusters were associated with both upper respiratory infection and wheezy bronchiolitis, while other strains were associated with upper respiratory infection alone. These results suggest that the present HRV-A isolates had a wide nucleotide divergence and were associated with acute respiratory infection, including upper respiratory infection and wheezy bronchiolitis, in Yamagata prefecture, Japan during the investigation period.
Assuntos
Análise por Conglomerados , Filogenia , Infecções por Picornaviridae/virologia , Infecções Respiratórias/virologia , Rhinovirus/classificação , Rhinovirus/isolamento & purificação , Adolescente , Criança , Pré-Escolar , Feminino , Humanos , Lactente , Japão , Masculino , Epidemiologia Molecular , Dados de Sequência Molecular , RNA Viral/genética , Rhinovirus/genética , Análise de Sequência de DNA , Homologia de Sequência de Aminoácidos , Homologia de Sequência do Ácido Nucleico , Proteínas Estruturais Virais/genéticaRESUMO
We report the case of a 2-y, 11-month-old boy with G1P[8] rotavirus infection accompanied by acute meningoencephalitis. Substitutions in both the VP4 and VP7 genes were found in the identified strain. A commonly circulating G1P[8] rotavirus with such mutations might be associated with the pathogenesis of CNS complications, including meningoencephalitis.
Assuntos
Meningite Viral/virologia , Infecções por Rotavirus/virologia , Rotavirus/classificação , Rotavirus/genética , Aciclovir/uso terapêutico , Antibacterianos/uso terapêutico , Antígenos Virais/genética , Antivirais/uso terapêutico , Povo Asiático , Proteínas do Capsídeo/genética , Carbenicilina/uso terapêutico , Pré-Escolar , Glicerol/uso terapêutico , Humanos , Japão/etnologia , Masculino , Meningite Viral/diagnóstico , FilogeniaRESUMO
Noroviruses (NVs) cause many cases of oyster- or clam-associated gastroenteritis in various countries. We collected 191 samples from Japanese oysters intended for raw consumption that had been harvested from the sea in two different areas between December 2001 and February 2002. To detect, quantitate, and phylogenetically analyze the NV genome in purified concentrates from the stomachs and digestive diverticula of these oysters, we amplified the NV capsid gene by reverse transcription-PCR. Phylogenetic analysis was performed by using the neighbor-joining method. We detected the NV genome in 17 of 191 oysters (9%). Phylogenetic analysis indicated genogroup I (Norwalk virus type) in 3 of the 17 oysters and genogroup II (Snow Mountain virus type) in the other 14. Both genogroups showed wide genetic diversity. To quantify the NV capsid gene in these oysters, we performed real-time PCR using genogroup-specific probes. More than 10(2) copies of the NV genome were detected in 11 of 17 oysters. The results suggested that about 10% of Japanese oysters intended for raw consumption harbored NVs, and more than 50% of those oysters in which NVs were detected had a large amount.