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1.
Artigo em Coreano | WPRIM (Pacífico Ocidental) | ID: wprim-816642

RESUMO

The 2019 novel coronavirus disease (COVID-19) outbreaks that emerged in Wuhan city, Hubei province, have led to a formidable number of confirmed cases that resulted in >5,700 deaths globally, including 143 countries in all 6 continents. The World Health Organization declared a Public Health Emergency of International Concern with a very high level of global risk assessment. Severe acute respiratory syndrome (SARS)-coronavirus-2 (SARS-CoV-2), the agent of COVID-19, has >79% nucleotide sequence homology to SARS-CoV; therefore, both belong to the genus betacoronavirus and subgenus sarbecovirus. The S1 domains of the two appeared to share the cellular receptor ACE2, but revealed a much higher S1-ACE2 binding affinity. As seen in many other human coronaviruses, SARS-CoV-2 also shows respiratory infection, but the basic reproductive number (R₀) in transmission and the clinical latency are quite dissimilar from those of SARS- or MERS-CoVs. Many scientists infer that the time point of cross-barrier transfer from bats to mediate animals or to humans should be a rather recent event based on the full-length genome analyses obtained from the very first patients. Copy-choice polymerization, which often leads to a significant genome recombination rate in most coronaviruses, predicts the continued emergence of novel coronaviruses.


Assuntos
Animais , Sequência de Bases , Quirópteros , Coronavirus , Surtos de Doenças , Emergências , Genoma , Humanos , Coronavírus da Síndrome Respiratória do Oriente Médio , Biologia Molecular , Polimerização , Polímeros , Saúde Pública , Recombinação Genética , Medição de Risco , Vírus da SARS , Síndrome Respiratória Aguda Grave , Organização Mundial da Saúde
2.
Annals of Dermatology ; : 122-129, 2020.
Artigo em Inglês | WPRIM (Pacífico Ocidental) | ID: wprim-811086

RESUMO

BACKGROUND: Loss-of-function mutations in the filaggrin gene (FLG), which encodes an epidermal protein crucial for the formation of a functional skin barrier, have been identified as a major predisposing factor in the etiopathogenesis of atopic dermatitis (AD). Recent reports of relatively low frequencies of FLG-null mutations among specific ethnic groups with AD necessitated analysis of the epigenetic regulation which may control FLG expression without altering its DNA sequence.OBJECTIVE: The study aimed to identify DNA methylation-dependent regulation of FLG expression.METHODS: Quantitative polymerase chain reaction was performed to determine the restoration of FLG mRNA expression in normal human epidermal keratinocyte (NHEK) cells after treatment with epigenetic modulating agents. Bisulfite genomic sequencing and pyrosequencing analyses of the FLG promoter region were conducted to identify the citical CpG sites relevant to FLG expression. We performed small-scale pilot study for epidermal tissues obtained from Korean patients with severe AD.RESULTS: We here show that DNA methylation in the FLG with non-CpG island promoter is responsible for the transcriptional regulation of FLG in undifferentiated NHEK cells. The methylation frequencies in a single CpG site of the FLG promoter were significantly higher in lesional epidermis than those in matched nonlesional epidermis of subjects with severe AD.CONCLUSION: Our in vitro and clinical studies point to this unique CpG site as a potential DNA methylation marker of FLG, which can be a promising therapeutic target in the complications of filaggrin-related skin barrier dysfunction as well as in AD.


Assuntos
Sequência de Bases , Causalidade , Dermatite Atópica , DNA , Metilação de DNA , Epiderme , Epigenômica , Grupos Étnicos , Expressão Gênica , Humanos , Técnicas In Vitro , Queratinócitos , Metilação , Projetos Piloto , Reação em Cadeia da Polimerase , Regiões Promotoras Genéticas , RNA Mensageiro , Pele
3.
Artigo em Inglês | WPRIM (Pacífico Ocidental) | ID: wprim-782227

RESUMO

For the past three decades, more than a thousand of genetic studies have been performed to find out the genetic variants responsible for the risk of asthma. Until now, all of the discovered single nucleotide polymorphisms have explained genetic effects less than initially expected. Thus, clarification of environmental factors has been brought up to overcome the ‘missing’ heritability. The most exciting solution is epigenesis because it intervenes at the junction between the genome and the environment. Epigenesis is an alteration of genetic expression without changes of DNA sequence caused by environmental factors such as nutrients, allergens, cigarette smoke, air pollutants, use of drugs and infectious agents during pre- and post-natal periods and even in adulthood. Three major forms of epigenesis are composed of DNA methylation, histone modifications, and specific microRNA. Recently, several studies have been published on epigenesis in asthma and allergy as a powerful tool for research of genetic heritability in asthma albeit epigenetic changes are at the starting point to obtain the data on specific phenotypes of asthma. In this presentation, we mainly review the potential role of DNA CpG methylation in the risk of asthma and its sub-phenotypes including nonsteroidal anti-inflammatory exacerbated respiratory diseases.


Assuntos
Poluentes Atmosféricos , Alérgenos , Aspirina , Asma , Sequência de Bases , Metilação de DNA , DNA , Epigenômica , Genoma , Código das Histonas , Hipersensibilidade , Metilação , MicroRNAs , Fenótipo , Polimorfismo de Nucleotídeo Único , Fumaça , Produtos do Tabaco
4.
Appl. cancer res ; 39: 1-7, 2019. ilustr.
Artigo em Inglês | LILACS (Américas), Inca | ID: biblio-1023627

RESUMO

Background: Mutations in the RAS/RAF pathway predict resistance to anti-epidermal growth factor receptor antibodies in colorectal cancer (CRC), and may be targets for future therapies. This study investigates concordance of BRAF, HRAS, KRAS, NRAS and PIK3CA mutation status in primary CRC with matched liver (n = 274), lung (n = 114) or combined liver and lung metastases (n = 14). Methods: Next generation sequencing was performed on DNA from formalin-fixed paraffin embedded CRC and matched liver and/or lung metastases, for recurrent mutations in BRAF, HRAS, KRAS, NRAS and PIK3CA and using the single-molecule molecular inversion probe method. Results: Paired sequencing results on all five genes were reached in 249 of the 402 cases (62%). The obtained number of unique reads was not always sufficient to confidently call the absence or presence of mutations for all regions of interest. The mutational status of matched pairs was highly concordant; 91.1% concordance for all five genes, 95.5% for KRAS, 99.1% for NRAS. Lung metastases more often harboured RAS mutations compared to liver metastases (71% vs. 48%, p < 0.001). Conclusions: In this large series of CRC we show that both primary tumors and corresponding metastases can be used to determine the mutational status for targeted therapy, given the high concordance rates. Next generation sequencing including a single molecule tags is feasible, however in combination with archival formalin-fixed paraffin embedded material is limited by coverage depth.


Assuntos
Humanos , Neoplasias Colorretais/genética , Proteínas ras/genética , Proteínas Proto-Oncogênicas B-raf/genética , Neoplasias Hepáticas/genética , Neoplasias Pulmonares/genética , Sequência de Bases , Neoplasias Colorretais/patologia , Sequenciamento de Nucleotídeos em Larga Escala , Neoplasias Hepáticas/secundário , Neoplasias Pulmonares/secundário , Mutação/genética
5.
Artigo em Inglês | WPRIM (Pacífico Ocidental) | ID: wprim-759593

RESUMO

BACKGROUND: The recent expansion of knowledge about various ABO alleles has led to the need for a comprehensive measure to cover the numerous polymorphisms dispersed in the ABO gene. A few studies have examined the diversity of the O allele compared to A or B subgroup alleles, resulting in antigenic changes. This study investigated the relationship between the serologic and molecular genetic characteristics of the O alleles in the Korean population. METHODS: One hundred and five samples from healthy blood group O subjects were selected randomly. The isoagglutinin titer was measured using a tube agglutination and gel microcolumn assay. The ABO alleles were analyzed by sequencing exons 6 and 7 of the ABO gene. When the origin of a heterozygous nucleotide sequence was ambiguous, it was separated into a single allele using mono-allele amplification or cloning. RESULTS: The median IgM isoagglutinin titer was eight. In contrast, the median IgG anti-A and anti-B isoagglutinin titers were 64 and 32, respectively. The IgG isoagglutinin titer showed a significant increase with age (P<0.0001). Six O alleles were observed in 105 blood group O populations by sequencing. The O01 and O02 alleles were common (0.57, 0.36). Three rare O alleles (O04, O05, and O06) and one novel non-deletional O allele were found. CONCLUSION: The distribution of isoagglutinin titers of blood group O and the genetic frequency of O alleles in this study would form the basis of the development and interpretation of ABO genotyping and serologic workup in the Korean population.


Assuntos
Aglutinação , Alelos , Sequência de Bases , Células Clonais , Clonagem de Organismos , Éxons , Imunoglobulina G , Imunoglobulina M , Biologia Molecular , Análise de Sequência
6.
Mycobiology ; : 120-125, 2019.
Artigo em Inglês | WPRIM (Pacífico Ocidental) | ID: wprim-760519

RESUMO

In 2017, small, elliptical, brownish purple spots on spears and ferns of asparagus were found in fields of Gangwon-do. The isolated fungal species was identified as an ascomycete Stemphylium vesicarium based on morphological characteristics and molecular phylogenic analyses including nucleotide sequences of the internal transcribed spacer (ITS), glyceraldehyde-3-phosphate dehydrogenase (GAPDH), and cytochrome b (cytb). A pathogenicity test revealed that S. vesicarium was the causal agent of purple spot disease on asparagus. The occurrence of purple spots caused by S. vesicarium on asparagus is the first report in Korea.


Assuntos
Ascomicetos , Sequência de Bases , Citocromos b , Gleiquênias , Coreia (Geográfico) , Oxirredutases , Virulência
7.
Artigo em Inglês | WPRIM (Pacífico Ocidental) | ID: wprim-742304

RESUMO

This study was undertaken to determine the complete mitochondrial DNA sequence and structure of the mitochondrial genome of Spirometra ranarum, and to compare it with those of S. erinaceieuropaei and S. decipiens. The aim of this study was to provide information of the species level taxonomy of Spirometra spp. using the mitochondrial genomes of 3 Spirometra tapeworms. The S. ranarum isolate originated from Myanmar. The mitochondrial genome sequence of S. ranarum was compared with that of S. erinaceieuropaei (GenBank no. KJ599680) and S. decipiens (Gen-Bank no. KJ599679). The complete mtDNA sequence of S. ranarum comprised 13,644 bp. The S. ranarum mt genome contained 36 genes comprising 12 protein-coding genes, 22 tRNAs and 2 rRNAs. The mt genome lacked the atp8 gene, as found for other cestodes. All genes in the S. ranarum mitochondrial genome are transcribed in the same direction and arranged in the same relative position with respect to gene loci as found for S. erinaceieuropaei and S. decipiens mt genomes. The overall nucleotide sequence divergence of 12 protein-coding genes between S. ranarum and S. decipiens differed by 1.5%, and 100% sequence similarity was found in the cox2 and nad6 genes, while the DNA sequence divergence of the cox1, nad1, and nad4 genes of S. ranarum and S. decipiens was 2.2%, 2.1%, and 2.6%, respectively.


Assuntos
Sequência de Bases , Cestoides , Classificação , DNA Mitocondrial , Genes vif , Genoma , Genoma Mitocondrial , Mianmar , RNA de Transferência , Spirometra
8.
Artigo em Inglês | WPRIM (Pacífico Ocidental) | ID: wprim-742302

RESUMO

In a population-based study with 4 years of follow up, we evaluated the prevalence of Coxiella burnetii in cattle on Ulleung Island, Korea. In this study, the rates of C. burnetii infection in cattle on Ulleung Island were determined by PCR and were found to be 0.3–1.0% in the period 2011–2014. All 17 C. burnetii partial 16S rRNA gene sequences from PCR-positive cattle were identical and 2 geographic representatives were included in our analysis. The nucleotide sequences of the 2 samples showed high (98.4–100%) identity with C. burnetii sequences obtained from the GenBank. In this long-term tracking study, the number of cattle positive for C. burnetii on Ulleung Island was low. To prevent the transmission of C. burnetii on Ulleung Island, control strategy should include biosecurity improvement in surveillance, livestock management, administering suitable tests before purchasing animals to detect C. burnetii shedders, and restricting movements between herds.


Assuntos
Animais , Sequência de Bases , Bovinos , Coxiella burnetii , Coxiella , Bases de Dados de Ácidos Nucleicos , Seguimentos , Genes de RNAr , Coreia (Geográfico) , Gado , Filogenia , Reação em Cadeia da Polimerase , Prevalência
9.
Artigo em Inglês | WPRIM (Pacífico Ocidental) | ID: wprim-741693

RESUMO

BACKGROUND/OBJECTIVES: Telomeres are located at the chromosomal ends and progressively shortened during each cell cycle. Telomerase, which is regulated by hTERT and c-MYC, maintains telomeric DNA sequences. Especially, telomerase is active in cancer and stem cells to maintain telomere length for replicative immortality. Recently we reported that walnut phenolic extract (WPE) can reduce cell viability in a colon cancer stem cell (CSC) model. We, therefore, investigated the effect of WPE on telomere maintenance in the same model. MATERIALS/METHODS: CD133+CD44+ cells from HCT116, a human colon cancer cell line, were sorted by Fluorescence-activated cell sorting (FACS) and treated with WPE at the concentrations of 0, 10, 20, and 40 µg/mL for 6 days. Telomere lengths were assessed by quantitative real-time PCR (qRT-PCR) using telomere specific primers and DNA extracted from the cells, which was further adjusted with single-copy gene and reference DNA (ddCt ). Telomerase activity was also measured by qRT-PCR after incubating the PCR mixture with cell protein extracts, which was adjusted with reference DNA (dCt ). Transcriptions of hTERT and c-MYC were determined using conventional RT-PCR. RESULTS: Telomere length of WPE-treated cells was significantly decreased in a dose-dependent manner (5.16 ± 0.13 at 0 µg/mL, 4.79 ± 0.12 at 10 µg/mL, 3.24 ± 0.08 at 20 µg/mL and 3.99 ± 0.09 at 40 µg/mL; P = 0.0276). Telomerase activities concurrently decreased with telomere length (1.47 ± 0.04, 1.09 ± 0.01, 0.76 ± 0.08, and 0.88 ± 0.06; P = 0.0067). There was a positive correlation between telomere length and telomerase activity (r = 0.9090; P < 0.0001). Transcriptions of both hTERT and c-MYC were also significantly decreased in the same manner. CONCLUSIONS: In the present cell culture model, WPE reduced telomere maintenance, which may provide a mechanistic link to the effect of walnuts on the viability of colon CSCs.


Assuntos
Sequência de Bases , Técnicas de Cultura de Células , Ciclo Celular , Linhagem Celular , Sobrevivência Celular , Colo , Neoplasias do Colo , DNA , Citometria de Fluxo , Humanos , Juglans , Fenol , Reação em Cadeia da Polimerase , Reação em Cadeia da Polimerase em Tempo Real , Células-Tronco , Telomerase , Telômero
10.
Artigo em Inglês | WPRIM (Pacífico Ocidental) | ID: wprim-758915

RESUMO

The clustered regularly interspaced short palindrome repeats (CRISPR)/CRISPR-associated protein 9 (Cas9) system is a versatile genome editing tool with high efficiency. A guide sequence of 20 nucleotides (nt) is commonly used in application of CRISPR/Cas9; however, the relationship between the length of the guide sequence and the efficiency of CRISPR/Cas9 in porcine cells is still not clear. To illustrate this issue, guide RNAs of different lengths targeting the EGFP gene were designed. Specifically, guide RNAs of 17 nt or longer were sufficient to direct the Cas9 protein to cleave target DNA sequences, while 15 nt or shorter guide RNAs had loss-of-function. Full-length guide RNAs complemented with mismatches also showed loss-of-function. When the shortened guide RNA and target DNA heteroduplex (gRNA:DNA heteroduplex) was blocked by mismatch, the CRISPR/Cas9 would be interfered with. These results suggested the length of the gRNA:DNA heteroduplex was a key factor for maintaining high efficiency of the CRISPR/Cas9 system rather than weak bonding between shortened guide RNA and Cas9 in porcine cells.


Assuntos
Sequência de Bases , Proteínas do Sistema Complemento , Sistemas CRISPR-Cas , DNA , Genoma , Nucleotídeos , RNA Guia , Suínos
11.
Artigo em Chinês | WPRIM (Pacífico Ocidental) | ID: wprim-774522

RESUMO

In order to understand the function of GDP-mannose pyrophosphorylase(GMPP) function and its regulation in polysaccharide biosynthesis mechanism in Dendrobium. D. huoshanense was used to clone GMPP gene. GMPP gene expression in D. huoshanense,D. officinale and D. moniliforme was also determined by qPCR. The results showed that the length of D. huoshanense GMPP gene c DNA sequence is 1 867 bp,containing 1 245 bp open reading frame(ORF),encoding 415 amino acids. Phylogenetic tree analysis showed that D. huoshanense,D. officinale and D. moniliforme are closely related with GMPP taken into consideration. Bioinformatics analysis demonstrated that GMPP sequence similarity among the three species reached as high as 99%. qPCR results indicated that GMPP genes was highly expressed in stem of D. huoshanense compared with its leaf,flower and root. According to GMPP gene expression profile in D. huoshanense,D. officinale and D. moniliforme grown in Huoshan area,it was clear that GMPP in D. huoshanense showed the highest expression level. Furthermore,our findings of GMPP gene expression profile will facilitate future researches into its polysaccharide biosynthetic mechanism.


Assuntos
Sequência de Bases , Clonagem Molecular , Dendrobium , Genética , Nucleotidiltransferases , Genética , Filogenia , Proteínas de Plantas , Genética , Polissacarídeos
12.
Artigo em Inglês | WPRIM (Pacífico Ocidental) | ID: wprim-772940

RESUMO

Enhancers activate transcription in a distance-, orientation-, and position-independent manner, which makes them difficult to be identified. Self-transcribing active regulatory region sequencing (STARR-seq) measures the enhancer activity of millions of DNA fragments in parallel. Here we used STARR-seq to generate a quantitative global map of rice enhancers. Most enhancers were mapped within genes, especially at the 5' untranslated regions (5'UTR) and in coding sequences. Enhancers were also frequently mapped proximal to silent and lowly-expressed genes in transposable element (TE)-rich regions. Analysis of the epigenetic features of enhancers at their endogenous loci revealed that most enhancers do not co-localize with DNase I hypersensitive sites (DHSs) and lack the enhancer mark of histone modification H3K4me1. Clustering analysis of enhancers according to their epigenetic marks revealed that about 40% of identified enhancers carried one or more epigenetic marks. Repressive H3K27me3 was frequently enriched with positive marks, H3K4me3 and/or H3K27ac, which together label enhancers. Intergenic enhancers were also predicted based on the location of DHS regions relative to genes, which overlap poorly with STARR-seq enhancers. In summary, we quantitatively identified enhancers by functional analysis in the genome of rice, an important model plant. This work provides a valuable resource for further mechanistic studies in different biological contexts.


Assuntos
Acetilação , Sequência de Bases , Desoxirribonuclease I , Metabolismo , Elementos Facilitadores Genéticos , Epigênese Genética , Genes de Plantas , Genômica , Métodos , Código das Histonas , Genética , Histonas , Metabolismo , Modelos Genéticos , Oryza , Genética , Regiões Promotoras Genéticas , Genética , Sequências Repetitivas de Ácido Nucleico , Genética , Análise de Sequência de DNA , Transcrição Genética
13.
Artigo em Chinês | WPRIM (Pacífico Ocidental) | ID: wprim-772011

RESUMO

OBJECTIVE@#To explore the clinical and genetic features of a patient suspected with Juvenile Parkinson's syndrome (JP).@*METHODS@#Clinical features of the patient were analyzed. Genomic DNA of the patient and his parents was extracted from peripheral blood samples and sequenced by exome capture sequencing. The nature and impact of detected mutations were predicted and validated.@*RESULTS@#The patient displayed typical features including resting tremor, bradykinesia, rigidity, but with excellent response to low dose levodopa. DNA sequencing showed that she has carried compound heterozygous mutations of the Parkin gene, namely c.1381dupC and c.619-1G>C, which were respectively inherited from his mother and father. Neither mutation was reported previously. Bioinformatic analysis predicted that both mutations are pathogenic.@*CONCLUSION@#The patient has JP caused by mutations of the Parkin gene. Exome capture sequencing is an accurate and efficient method for genetic diagnosis of such disease.


Assuntos
Adolescente , Sequência de Bases , Feminino , Humanos , Mutação , Doença de Parkinson , Ubiquitina-Proteína Ligases , Sequenciamento Completo do Exoma
14.
Artigo em Chinês | WPRIM (Pacífico Ocidental) | ID: wprim-772007

RESUMO

OBJECTIVE@#To detect potential variant of AR gene in an infant with complete androgen insensitivity syndrome.@*METHODS@#The coding regions and splicing sites of the AR gene were subjected to PCR amplification and direct DNA sequencing. Fluorescence quantitative PCR was also used to detect copy number alterations of exons 2 to 8 of the AR gene.@*RESULTS@#Deletion of exons 2 to 8 was detected in the proband, and the results were verified among the family members.@*CONCLUSION@#Hemizygotic deletion of exons 2 to 8 of the AR gene probably underlies the complete androgen insensitivity syndrome in this infant.


Assuntos
Síndrome de Resistência a Andrógenos , Genética , Sequência de Bases , Éxons , Humanos , Lactente , Masculino , Reação em Cadeia da Polimerase , Receptores Androgênicos , Genética
15.
Artigo em Chinês | WPRIM (Pacífico Ocidental) | ID: wprim-771954

RESUMO

OBJECTIVE@#To identify mutation of the PAX6 gene in a patient with congenital aniridia.@*METHODS@#DNA was extracted from peripheral blood sample of the patient and analyzed by direct PCR-Sanger sequencing.@*RESULTS@#The proband was found to harbor a heterozygous c.239T>A (p.Ile80Asn) mutation of the PAX6 gene. The same mutation was not found in his parents and 150 healthy controls.@*CONCLUSION@#A novel mutation of the PAX6 gene has been identified in a sporadic case with congenital aniridia.


Assuntos
Aniridia , Genética , Sequência de Bases , Humanos , Mutação , Fator de Transcrição PAX6 , Genética , Linhagem
16.
Artigo em Chinês | WPRIM (Pacífico Ocidental) | ID: wprim-771929

RESUMO

OBJECTIVE@#To explore the potential pathogenetic mutations of primary hypereosinophilia(HEN)by sequencing FGFR1 FLT3, MPL and JAK2 genes, and to clarify their effect on clinical manifestation and prognosis of HEN patients.@*METHODS@#The direct DNA sequencing was employed to detect the gene mutations of FGFR1, FLT3, MPL and JAK2 in HEN patients.@*RESULTS@#One deletion mutation (2654_2753del) within tyrosine kinase domain of FLT3 gene was found in a patient suffered from severe symptoms and ended with dismal outcome, which induced a premature stop codon (G885fsX888). For FGFR1, a new variation described as 1014_1019del AACAGT for nucleotide change was found in 19 cases, resulting in T339_V340del at the protein level.@*CONCLUSION@#The deletion of 6 bases in the FGFR1 gene (1014_1019del AACAGT) is first reported as non-synonymous SNP (nsSNP) site in the patients with primary hypereosinophilia. Deletion mutations in the FLT3 gene may be related with malignant clinical features and poor prognosis.


Assuntos
Sequência de Bases , Humanos , Síndrome Hipereosinofílica , Genética , Mutação , Receptores de Trombopoetina , Deleção de Sequência , Tirosina Quinase 3 Semelhante a fms
17.
Artigo em Inglês | WPRIM (Pacífico Ocidental) | ID: wprim-764239

RESUMO

Canine adenovirus type 1 (CAV-1) infection results in hepatitis in dogs. In this study, we investigated the biologic and genetic characteristics of the CAV-1 vaccine strain (CAV1V) to improve quality control about CAV vaccine. The identity of CAV1V as CAV-1 was confirmed based on its cytopathic effects and the results of hemagglutination (HA) and immunofluorescence assays, and electron microscopy. The CAV1V strain reached 10(7.5) TCID(50)/mL in MDCK cells at 4 days post-inoculation and exhibited hemmagglutination activity of 256 U using guinea pig erythrocytes. Intranuclear fluorescence in the infected cells was observed and typical adenoviruses were observed in electon microscope. CAV1V strain was identified as a CAV-1 strain by nucleotide sequence analysis. In a comparison of the nucleotide sequences of the fiber genes of several CAV strains, CAV1V showed the highest similarity (99.8%) with the GLAXO strain, which was isolated in Canada. Our biological characterization of CAV1V will facilitate quality control of the canine hepatitis vaccine.


Assuntos
Adenoviridae , Adenovirus Caninos , Animais , Sequência de Bases , Canadá , Cães , Eritrócitos , Fluorescência , Imunofluorescência , Cobaias , Hemaglutinação , Hepatite , Células Madin Darby de Rim Canino , Microscopia Eletrônica , Controle de Qualidade
18.
Artigo em Coreano | WPRIM (Pacífico Ocidental) | ID: wprim-764232

RESUMO

Noroviruses (NoV) are the major viral pathogen causing epidemic acute gastroenteritis and outbreaks of foodborne and waterborne illness. During the local festival in Chungnam province, group food poisoning occurred outbreak by NoV infections in Jan 2019. In this study, epidemiological analysis and molecular characterization were conducted such as genotyping, phylogeny. The prevalent genotypes of food poisoning events were NoV GII.3 and GII.17, and NoV GII.3 and GII.17 isolates of this study were completely matched in nucleotide sequence comparison of capsid gene region, respectively. In underground water and stream water, various multiple genotypes of noroviruses were detected including NoV GII.3, GII.8 and GI.4 in aquatic environment of the local festival site. Among 32 worker samples, various NoVs of five genotypes (GI.7, GI.8, GII.3, GII.8, GII.17) were detected in 12 samples and expected to causing NoV contaminated by exposure to groundwater. NoV genotype GII.3, which was detected from groundwater 2, was completely consistent with that of patients and workers. Therefore, groundwater within the local festival site could be main cause of food poisoning event. Because NoV outbreaks are caused by fecal to oral transmission, proper management of sewage purification facilities, groundwater and sanitary toilets is required for many visitors, and efforts are needed to maintain clean environment.


Assuntos
Sequência de Bases , Capsídeo , Surtos de Doenças , Estudos Epidemiológicos , Doenças Transmitidas por Alimentos , Gastroenterite , Genótipo , Água Subterrânea , Férias e Feriados , Humanos , Coreia (Geográfico) , Norovirus , Filogenia , Rios , Esgotos , Água
19.
Artigo em Inglês | WPRIM (Pacífico Ocidental) | ID: wprim-763368

RESUMO

PURPOSE: Canine influenza virus (CIV), H3N2, carries potentiality for zoonotic transmission and genetic assortment which raises a concern on possible epidemics, and human threats in future. To manage possible threats, the development of rapid and effective methods of CIV vaccine production is required. The plant provides economical, safe, and robust production platform. We investigated whether hemagglutinin (HA) antigen from Korea-originated CIV could be produced in Nicotiana benthamiana and lettuce, Lactuca sativa by a DNA viral vector system. MATERIALS AND METHODS: We used DNA sequences of the HA gene from Korean CIV strain influenza A/canine/Korea/S3001/2015 (H3N2) for cloning into a geminiviral expression vectors to express recombinant HA (rHA) antigen in the plant. Agrobacterium-mediated infiltration was performed to introduce HA-carrying vector into host plants cells. Laboratory-grown N. benthamiana, and grocery-purchased or hydroponically-grown lettuce plant leaves were used as host plants. RESULTS: CIV rHA antigen was successfully expressed in host plant species both N. benthamiana and L. sativa by geminiviral vector. Both complex-glycosylated and basal-glycosylated form of rHA were produced in lettuce, depending on presence of endoplasmic reticulum (ER) retention signal. In terms of rHA expression level, canine HA (H3N2) showed preference to the native signal peptide than ER retention signal peptide in the tested geminiviral vector system. CONCLUSION: Grocery-purchased lettuce leaves could serve as an instant host system for the transient expression of influenza antigen at the time of emergency. The geminiviral vector was able to induce expression of complex-glycosylated and basal-glycosylated rHA in lettuce and tobacco.


Assuntos
Sequência de Bases , Células Clonais , Clonagem de Organismos , DNA , Emergências , Retículo Endoplasmático , Hemaglutininas , Humanos , Influenza Humana , Alface , Orthomyxoviridae , Folhas de Planta , Plantas , Sinais Direcionadores de Proteínas , Tabaco
20.
Artigo em Chinês | WPRIM (Pacífico Ocidental) | ID: wprim-776813

RESUMO

OBJECTIVE@#To identify pathogenic variations of EXT1 and EXT2 genes in two Chinese pedigrees affected with hereditary multiple exostosis (HME).@*METHODS@#Genomic DNA was extracted from peripheral blood samples using a phenol-chloroform method. PCR and Sanger sequencing was conducted to amplify the exons and the flanking intronic regions of the EXT1 and EXT2 genes.@*RESULTS@#DNA sequencing has revealed a heterozygous missense variation c.812A>G (p.Tyr271Cys) in the exon 1 of EXT1 in pedigree 1, and a heterozygous frameshift variation c.1431dup (p.Ser478Leufs*43) in the exon 6 of EXT1 in the proband from pedigree 2. Both variations have co-segregated with the disease phenotype, which was also consistent with previous report.@*CONCLUSION@#Two heterozygous pathogenic variations underlying HME have been identified. The result has facilitated genetic counseling and prenatal diagnosis for the affected pedigrees.


Assuntos
Grupo com Ancestrais do Continente Asiático , Sequência de Bases , Análise Mutacional de DNA , Exostose Múltipla Hereditária , Genética , Patologia , Mutação da Fase de Leitura , Humanos , Mutação de Sentido Incorreto , N-Acetilglucosaminiltransferases , Genética , Linhagem
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