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OBJECTIVES: Macrolide-resistant Bordetella pertussis (MRBP) has been emerging and prevailing in mainland China since 2011. In this study, we aimed to investigate the genotype and macrolide resistance of circulating B. pertussis in East and Southeast Asia using genetic analyses. METHODS: A total of 302 DNA extracts from clinical specimens and isolates from 2010 to 2020 were analyzed: 145 from Vietnam, 76 from Cambodia, 48 from Taiwan, and 33 from Japan. Genotypes were determined by multilocus variable-number tandem-repeat analysis (MLVA). Macrolide-resistant A2047G mutation in B. pertussis 23S rRNA was investigated using the duplex Cycleave real-time polymerase chain reaction (PCR) assay. Whole-genome sequencing was performed on two MRBP isolates that were identified for the first time in Taiwan. RESULTS: Overall, 286 DNA extracts (95%) generated a complete MLVA genotype and 283 DNA extracts (94%) yielded a complete result for the A2047G mutation analysis. The A2047G mutation was detected in 18 DNA extracts: fourteen from Vietnam, one from Cambodia, two from Taiwan, and one from Japan. Most of them (78%) showed the genotypes MT104 and MT195, which have previously been reported in Chinese MRBP isolates. Further, the Taiwanese MRBP isolates were classified into the MT104 clade of Chinese MRBP isolates. CONCLUSION: After MRBP emerged and spread in mainland China, it may have spread to East and Southeast Asia in the 2010s. Continued surveillance targeting the A2047G mutation of MRBP is needed to prevent further spread of this emerging pathogen.
Assuntos
Bordetella pertussis , Coqueluche , Humanos , Bordetella pertussis/genética , Macrolídeos/farmacologia , Coqueluche/epidemiologia , Antibacterianos/farmacologia , Genótipo , Farmacorresistência Bacteriana , Mutação , Sudeste Asiático , Ásia OrientalRESUMO
Multilocus variable-number tandem repeat analysis (MLVA) is widely used for genotyping of Bordetella pertussis, the causative bacteria for pertussis. However, MLVA genotyping is losing its discriminate power because prevalence of the epidemic MT27 strain (MLVA-27) is increasing worldwide. To address this, we developed a single nucleotide polymorphism (SNP) genotyping method for MT27 based on multiplexed single-base extension (SBE) assay. A total of 237 MT27 isolates collected in Japan during 1999-2018 were genotyped and classified into ten SNP genotypes (SG1 to SG10) with a Simpson's diversity index (DI) of 0.79 (95% CI 0.76-0.82). Temporal trends showed a marked increase in the genotypic diversity in the 2010s: Simpson's DI was zero in 1999-2004, 0.16 in 2005-2009, 0.83 in 2010-2014, and 0.76 in 2015-2018. This indicates that the SNP genotyping is applicable to the recently circulating MT27 strain. Additionally, almost all outbreak-associated MT27 isolates were classified into the same SNP genotypes for each outbreak. Multiplexed SBE assay allows for rapid and simple genotyping, indicating that the SNP genotyping can potentially be a useful tool for subtyping the B. pertussis MT27 strain in routine surveillance and outbreak investigations.
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Bordetella pertussis/genética , DNA Bacteriano/genética , Técnicas de Genotipagem , Tipagem de Sequências Multilocus , Polimorfismo de Nucleotídeo Único , Humanos , Japão/epidemiologia , Coqueluche/epidemiologia , Coqueluche/genéticaRESUMO
BACKGROUND: The Taiwanese national 23-valent pneumococcal polysaccharide vaccine (PPV23) program in adults ≥75 years of age and the 13-valent pneumococcal conjugate vaccine (PCV13) program for children were implemented in 2008 and 2013, respectively. In this study we evaluated PPV23 vaccine effectiveness (PPV23VE) in the elderly, with regard to both direct protection from the vaccine itself and the indirect protection conferred by PCV13 immunization in children. METHODS: The incidence of invasive pneumococcal disease (IPD) in Taiwan from July 2008 to June 2016 was collected from IPD surveillance data. A comparison of IPD incidence with a nationwide vaccination registry allowed an estimation of PPV23VE by the screening and indirect cohort methods. RESULTS: The incidence of IPD in adults ≥75 years of age ranged from 13.9 per 100,000 inhabitants during the period July 2008-June 2013 to 10.4 per 100,000 inhabitants between July 2013 and June 2016 (relative risk [RR]: 0.75; 95% confidence interval [95% CI]: 0.67-0.85). According to the screening method, PPV23VE against death within 30 days of IPD onset, all IPD, and PPV23-serotype IPD was 32.5% (95% CI: 17.5-44.7%), 33.9% (95% CI: 25.2-41.5%) and 43.4% (95% CI: 34.4-51.2%), respectively. PPV23VE with the indirect cohort method was 39.0% (95% CI: 15.5-55.9%) for all PPV23 serotypes and 71.5% (95% CI: 44.2-85.4%) for 11 serotypes included in PPV23 but not in PCV13. During the period July 2008-June 2012, PPV23VE against PPV23-serotype IPD was 55.1% (95% CI: 27.2-72.3%). CONCLUSIONS: PPV23 is able to prevent IPD and 30-day fatality in adults 75 years of age and older due to a combination of direct effects from PPV23 and indirect effects from PCV13. It might confer higher protection against PPV23-serotype IPD before the introduction of PCV13 program in children.
Assuntos
Infecções Pneumocócicas/prevenção & controle , Vacinas Pneumocócicas/administração & dosagem , Streptococcus pneumoniae/imunologia , Adolescente , Adulto , Idoso , Idoso de 80 Anos ou mais , Criança , Pré-Escolar , Estudos de Coortes , Feminino , Humanos , Programas de Imunização , Masculino , Pessoa de Meia-Idade , Infecções Pneumocócicas/epidemiologia , Infecções Pneumocócicas/microbiologia , Avaliação de Programas e Projetos de Saúde , Sistema de Registros , Streptococcus pneumoniae/classificação , Streptococcus pneumoniae/genética , Taiwan/epidemiologia , Adulto JovemRESUMO
Purpose. Human-adapted Bordetella parapertussis is one of the causative agents of whooping cough; however, there are currently no genotyping systems with high discriminatory power for this bacterial pathogen. We therefore aimed to develop a multilocus variable-number tandem repeat analysis (MLVA) for human-adapted B. parapertussis.Methodology. Four highly polymorphic variable number tandem repeat (VNTR) loci in the B. parapertussis genome were selected and amplified by multiplex PCR. MLVA was performed based on the number of tandem repeats at VNTR loci. The discriminatory power of MLVA was evaluated with three laboratory reference strains and 50 human isolates of B. parapertussis.Results. Multiplex PCR-based MLVA characterized 53 B. parapertussis reference strains and isolates into 25 MLVA types and the Simpson diversity index was 0.91 (95 % confidence interval, 0.86-0.97). The three reference strains exhibited different MLVA types. Thirty-one Japanese isolates, ten French isolates and three Taiwanese isolates belonged to fourteen, nine and three MLVA types, respectively. In contrast, all five Australian isolates belonged to the same type. Two Japanese isolates collected from patients with known epidemiological links had the same type.Conclusion. Our novel MLVA method has high discriminatory power for genotyping human B. parapertussis. Regarding this organism, this genotyping system is a promising tool for epidemiological surveillance and investigating outbreaks.
Assuntos
Bordetella parapertussis/genética , Bordetella parapertussis/isolamento & purificação , Tipagem de Sequências Multilocus/métodos , Coqueluche/microbiologia , Bordetella parapertussis/classificação , Humanos , Repetições Minissatélites , Coqueluche/diagnósticoRESUMO
BACKGROUND: Streptococcus pneumoniae infections in Taiwan mostly occur in children aged 2-4 years. Because of a significant increase in the incidence of serotype 19A-related infections, the 13-valent pneumococcal conjugate vaccine (PCV13) was initially introduced in the national immunization program for children 2-5 years of age, prior to the national programs for infants. We have assessed the impact of such vaccination programs in reducing the incidence of invasive pneumococcal disease (IPD) in Taiwanese children. METHODS: We analyzed the national data on IPDs from the Taiwan Centers for Disease Control between 2008 and 2017. We calculated the incidence rates of IPD and incidence rate ratios (IRRs) between years for different serotypes to estimate the effectiveness of the vaccination programs. RESULTS: The national catch-up primary vaccination schedule successfully reduced the incidence rate of IPD from 17.8/100 000 in 2012 to 5.5/100 000 in 2017 among children aged 0-5 years. The IRR (2017 over 2012) was 0.31, corresponding to a 69% reduction. A modest herd effect was also observed, with a 37% reduction in the incidence of IPD in elderly people (≥70 years) from 2012 to 2017. The incidence of IPD caused by serotype 19A in children aged 0-5 years was reduced by 32.6-44.3% yearly from 2012 to 2017. In 2015, serogroup 15 outnumbered 19A, to become the leading serotypes in children 0-5 years old. CONCLUSIONS: Special catch-up vaccination programs starting from children 2-5 years of age with PCV13 have been highly effective in reducing the incidence of IPD, especially as caused by serotype 19A, in Taiwanese children.
Assuntos
Vacinas Pneumocócicas/uso terapêutico , Streptococcus pneumoniae/imunologia , Streptococcus pneumoniae/patogenicidade , Vacinas Conjugadas/uso terapêutico , Pré-Escolar , Feminino , Humanos , Masculino , Sorogrupo , Taiwan , VacinaçãoRESUMO
To guard genome integrity, response mechanisms coordinately execute the G2/M checkpoint in responding to stress. p38 MAPK is activated to prolong the G2 phase for completion of damage repair. Tlk activity is required for DNA repair, chromosome segregation and G2 recovery. However, the involvement of Tlk in G2 recovery differs from previous findings that Tlk overexpression delays the G2/M transition. To clarify this difference, genetic interaction experiments were performed using the second mitotic wave as model system. The results indicate that Tlk overexpression prolongs the G2 phase through p38 MAPK activation, independent of Tlk kinase activity. The results of co-immunoprecipitation, database search and RNAi screening suggest that eEF1α1 and Hsc70-5 links Tlk to Tak1. Reduced gene activities of Tlk, Hsc70-5, eEF1α1 and/or Tak1 couldn't prolong the G2 phase induced by heat shock, indicating that these proteins work together to elevate p38 MAPK activity. In contrast, a high level of wild type Tlk decreases phosphorylated p38 MAPK levels. Thus, the difference is explained by a dual function of Tlk. When under stress, inactive Tlk increases p38 MAPK activity to prolong the G2 phase, and then activated Tlk modulates activities of p38 MAPK and Asf1 to promote G2 recovery afterwards.
Assuntos
Proteínas de Drosophila/metabolismo , MAP Quinase Quinase Quinases/metabolismo , Proteínas Serina-Treonina Quinases/metabolismo , Proteínas Quinases p38 Ativadas por Mitógeno/metabolismo , Animais , Animais Geneticamente Modificados/metabolismo , Drosophila/metabolismo , Proteínas de Drosophila/antagonistas & inibidores , Proteínas de Drosophila/genética , Embrião não Mamífero/metabolismo , Olho/citologia , Olho/metabolismo , Feminino , Pontos de Checagem da Fase G2 do Ciclo Celular , Regulação da Expressão Gênica no Desenvolvimento , Proteínas de Choque Térmico HSC70/antagonistas & inibidores , Proteínas de Choque Térmico HSC70/genética , Proteínas de Choque Térmico HSC70/metabolismo , MAP Quinase Quinase Quinases/antagonistas & inibidores , MAP Quinase Quinase Quinases/genética , Mutagênese , Fosforilação , Proteínas Serina-Treonina Quinases/genética , Interferência de RNA , RNA Interferente Pequeno/metabolismo , Proteínas Quinases p38 Ativadas por Mitógeno/genéticaRESUMO
The use of pneumococcal vaccine plays an important role for prevention of invasive pneumococcal disease (IPD). However, introducing the pneumococcal vaccine into the national immunization program (NIP) is complex and costly. The strategy of progressively integrating the pneumococcal conjugate vaccine (PCV) into the NIP in Taiwan provides valuable experience for policy makers. The 7-valent PCV (PCV7) was first available in Taiwan in late 2005. PCV7 was first provided free to children with underlying diseases, those in vulnerable socioeconomic status, and those with inadequate health care resources. The catch-up immunization program with the 13-valent PCV was launched in 2013 and the national pneumococcal immunization program was implemented in 2015. Children aged 2-5 years had the highest incidence of IPD among pediatric population in Taiwan. Although the incidence of IPD caused by PCV7 serotypes has declined, the overall incidence of IPD remained high in the context of PCV7 use in the private sector. A surge of IPD caused by serotype 19A occurred, accounting for 53.6% of IPD cases among children aged ≤ 5 years in 2011-2012. After the implementation of the national pneumococcal immunization program, serogroup 15 has become the leading serogroup for IPD in children. Continued surveillance is necessary to monitor the serotype epidemiology in Taiwan.
RESUMO
BACKGROUND: Seven-valent pneumococcal conjugate vaccine (PCV) has been available in Taiwan since late 2005. A national catch-up program was launched in Taiwan in 2013, providing 1 dose of 13-valent PCV to children aged 2-5 years. Here, we report the epidemiology of invasive pneumococcal disease (IPD) in children aged ≤5 years in this setting. METHODS: We collected demographic and clinical information for pediatric patients (≤5 years) with IPD between 2008 and 2013. The incidence of IPD was estimated. The logs for PCV import into Taiwan were obtained to evaluate the impact of PCV usage on IPD epidemiology. RESULTS: The overall incidence of IPD in children aged ≤5 years was 15.9 cases per 100,000 person-years. The IPD incidence caused by 7-valent PCV serotypes decreased significantly from 10.0 cases per 100,000 person-years in 2008 to 2.3 cases per 100,000 person-years in 2013. The incidence of IPD caused by serotype 19A increased substantially from 1.7 cases per 100,000 person-years in 2008 to 10.3 cases per 100,000 person-years in 2012, followed by a significant decrease to 5.6 cases per 100,000 person-years in 2013. The significant decrease in the incidence of serotype 19A IPD occurred primarily in children aged 2-5 years. CONCLUSIONS: The 13-valent PCV catch-up program was associated with a significant decrease in serotype 19A IPD incidence in 2013, primarily in children eligible for the 13-valent PCV immunization. Continued surveillance is necessary to assess the further impact of the national catch-up program on pediatric IPD epidemiology in Taiwan.
Assuntos
Infecções Pneumocócicas/epidemiologia , Infecções Pneumocócicas/prevenção & controle , Vacinas Pneumocócicas/imunologia , Vacinação , Vacinas Conjugadas/imunologia , Pré-Escolar , Feminino , Vacina Pneumocócica Conjugada Heptavalente , Humanos , Incidência , Lactente , Recém-Nascido , Masculino , Programas Nacionais de Saúde , Sorogrupo , Streptococcus pneumoniae/classificação , Streptococcus pneumoniae/imunologia , Taiwan/epidemiologiaRESUMO
We report 2 cases of neonatal Legionella infection associated with aspiration of contaminated water used in hospitals to make infant formula. The molecular profiles of Legionella strains isolated from samples from the infants and from water dispensers were indistinguishable. Our report highlights the need to consider nosocomial legionellosis among neonates who have respiratory symptoms.
Assuntos
Infecção Hospitalar , Fórmulas Infantis , Legionella/isolamento & purificação , Legionelose/diagnóstico , Legionelose/microbiologia , Microbiologia da Água , Humanos , Recém-Nascido , Legionella/classificação , Legionella/genética , Legionelose/epidemiologia , Masculino , Vigilância da População , Taiwan/epidemiologiaRESUMO
In this study, TaqMan fluorescent quantitative real-time PCR was performed to quantify Legionella species in reservoirs. Water samples were collected from 19 main reservoirs in Taiwan, and 12 (63.2%) were found to contain Legionella spp. The identified species included uncultured Legionella spp., L. pneumophila, L. jordanis, and L. drancourtii. The concentrations of Legionella spp. and L. pneumophila in the water samples were in the range of 1.8×10(2)-2.6×10(3) and 1.6×10(2)-2.4×10(2) cells/L, respectively. The presence and absence of Legionella spp. in the reservoir differed significantly in pH values. These results highlight the importance that L. pneumophila, L. jordanis, and L. drancourtii are potential pathogens in the reservoirs. The presence of L. pneumophila in reservoirs may be a potential public health concern that must be further examined.
Assuntos
Água Potável/microbiologia , Legionella/crescimento & desenvolvimento , Microbiologia da Água , Monitoramento Ambiental/métodos , Corantes Fluorescentes , Legionella/classificação , Legionella/isolamento & purificação , Reação em Cadeia da Polimerase em Tempo Real , TaiwanRESUMO
INTRODUCTION: Serotype replacement after the introduction of seven-valent pneumococcal conjugate vaccine (PCV7) and the future availability of multivalent PCVs prompted the listing of invasive pneumococcal disease (IPD) as a notifiable disease in Taiwan in October 2007. Here, we report the national surveillance results. METHODS: The study population comprised the whole nation of Taiwan from 2008 to 2012. Restricting to cases with viable isolates, we calculated the incidence, case fatality ratio, prevalence of serotype 19A, and percentage of vaccine preventable IPD. RESULTS: 3659 cases of IPD were identified yielding an incidence of 3.2 per 100,000 population; the highest incidence was among children aged 2-4 years (21.1 per 100,000 population). The case fatality ratio was 9.2% and the highest ratio was among adults aged ≥75 years (19.0%). The percentage of PCV7 preventable IPD decreased for all age groups, especially sharply among children aged 2-4 years, from 65.8% in 2008 to 12.9% in 2012. The prevalence of serotype 19A increased from 5.5% in 2008 to 25.3% in 2012 among all Streptococcus pneumoniae, displaying a differential temporal emergence among different age groups. Serotype 19A became the most prevalent serotype among children aged <2 years in 2009, children aged 2-4 and 5-17 years in 2010, and adults aged 18-49 years in 2012. CONCLUSIONS: The incidence of IPD fluctuated during the study period, with ongoing decrease due to PCV7 vaccine serotypes and increase due to non-vaccine serotypes. Serotype 19A became the most prevalent serotype in 2010 among all S. pneumoniae.
Assuntos
Infecções Pneumocócicas/epidemiologia , Vigilância de Evento Sentinela , Streptococcus pneumoniae/classificação , Adolescente , Adulto , Idoso , Criança , Pré-Escolar , Notificação de Doenças , Humanos , Incidência , Lactente , Pessoa de Meia-Idade , Infecções Pneumocócicas/mortalidade , Sorotipagem , Taiwan/epidemiologia , Adulto JovemRESUMO
Bordetella pertussis causes pertussis, a respiratory disease that is most severe for infants. Vaccination was introduced in the 1950s, and in recent years, a resurgence of disease was observed worldwide, with significant mortality in infants. Possible causes for this include the switch from whole-cell vaccines (WCVs) to less effective acellular vaccines (ACVs), waning immunity, and pathogen adaptation. Pathogen adaptation is suggested by antigenic divergence between vaccine strains and circulating strains and by the emergence of strains with increased pertussis toxin production. We applied comparative genomics to a worldwide collection of 343 B. pertussis strains isolated between 1920 and 2010. The global phylogeny showed two deep branches; the largest of these contained 98% of all strains, and its expansion correlated temporally with the first descriptions of pertussis outbreaks in Europe in the 16th century. We found little evidence of recent geographical clustering of the strains within this lineage, suggesting rapid strain flow between countries. We observed that changes in genes encoding proteins implicated in protective immunity that are included in ACVs occurred after the introduction of WCVs but before the switch to ACVs. Furthermore, our analyses consistently suggested that virulence-associated genes and genes coding for surface-exposed proteins were involved in adaptation. However, many of the putative adaptive loci identified have a physiological role, and further studies of these loci may reveal less obvious ways in which B. pertussis and the host interact. This work provides insight into ways in which pathogens may adapt to vaccination and suggests ways to improve pertussis vaccines. IMPORTANCE Whooping cough is mainly caused by Bordetella pertussis, and current vaccines are targeted against this organism. Recently, there have been increasing outbreaks of whooping cough, even where vaccine coverage is high. Analysis of the genomes of 343 B. pertussis isolates from around the world over the last 100 years suggests that the organism has emerged within the last 500 years, consistent with historical records. We show that global transmission of new strains is very rapid and that the worldwide population of B. pertussis is evolving in response to vaccine introduction, potentially enabling vaccine escape.
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Bordetella pertussis/classificação , Bordetella pertussis/genética , Vacina contra Coqueluche/imunologia , Vacinação/métodos , Coqueluche/epidemiologia , Coqueluche/microbiologia , Adaptação Biológica , Bordetella pertussis/imunologia , Bordetella pertussis/isolamento & purificação , Análise por Conglomerados , Doenças Transmissíveis Emergentes/epidemiologia , Doenças Transmissíveis Emergentes/microbiologia , Evolução Molecular , Genoma Bacteriano , Saúde Global , Humanos , Lactente , Vacina contra Coqueluche/administração & dosagem , FilogeniaRESUMO
The aim of the present study was to investigate the epidemiology of Legionnaires' disease (LD) caused by Legionella longbeachae in Taiwan during 2006-2010. A total of six cases were identified prospectively, accounting for 1.6% of all laboratory-confirmed LD cases and 4.4% of culture-positive LD cases. All six cases occurred between April and August. The male to female ratio was 0.5. These six LD patients had a higher median age than those with LD due to Legionella pneumophila. Four of the six patients presented with pleural effusion and five survived the infection episode. Only two patients had a potential soil contact history prior to LD onset. The patients resided in divergent geographical areas without a common exposure history. The individual genomic DNA banding patterns of the six L. longbeachae isolates analyzed by pulsed-field gel electrophoresis (PFGE) were unique, supporting the hypothesis that the L. longbeachae infections occurred sporadically.
Assuntos
Legionella longbeachae/isolamento & purificação , Legionelose/epidemiologia , Adulto , Idoso , Idoso de 80 Anos ou mais , DNA Bacteriano/genética , Eletroforese em Gel de Campo Pulsado , Feminino , Humanos , Legionella longbeachae/genética , Legionelose/microbiologia , Masculino , Pessoa de Meia-Idade , Derrame Pleural , Estudos Prospectivos , Taiwan/epidemiologiaRESUMO
BACKGROUND: The soil-dwelling pathogen Burkholderia pseudomallei is the cause of melioidosis. In this study, the geographical and temporal distributions of predominant molecular patterns occurring in clinical and environmental isolates of B. pseudomallei were characterised. METHODS: A collection of 194 human and 59 soil B. pseudomallei isolates obtained in Taiwan were analysed by pulsed-field gel electrophoresis (PFGE) and multilocus sequence typing (MLST). RESULTS: Five predominant PFGE types (VI, X, XI, XV and XVI) among human isolates were identified between 2004 and 2010. Among them, types VI, X and XI correspond to ST58, and types XV and XVI correspond to ST99. The distribution of B. pseudomallei with distinct PFGE or MLST types was clustered in different towns in southern Taiwan. Clusters of B. pseudomallei have successively appeared in the town of Jiading, which is located in the Er-Ren River Basin of southern Taiwan. CONCLUSIONS: Burkholderia pseudomallei isolates were geographically and temporally clustered in Taiwan. This finding supports the contention that the Er-Ren River Basin now constitutes the highest risk area for melioidosis in Taiwan.
Assuntos
Burkholderia pseudomallei/genética , Melioidose/microbiologia , Microbiologia do Solo , Técnicas de Tipagem Bacteriana , Burkholderia pseudomallei/classificação , Burkholderia pseudomallei/isolamento & purificação , Sistemas de Informação Geográfica , Humanos , Melioidose/epidemiologia , Tipagem de Sequências Multilocus , Taiwan/epidemiologiaRESUMO
A survey for the prevalence if Burkholderia pseudomallei in soil in Taiwan found that its incidence is comparable to that in other regions of the world where melioidosis is endemic. The presence of identical genetic patterns among the clinical and environmental isolates evaluated suggested a link between the pathogens present in contaminated soil and the emergence of indigenous melioidosis.
Assuntos
Burkholderia pseudomallei/isolamento & purificação , Melioidose/epidemiologia , Microbiologia do Solo , Técnicas de Tipagem Bacteriana , Burkholderia pseudomallei/classificação , Burkholderia pseudomallei/genética , Análise por Conglomerados , Impressões Digitais de DNA , Eletroforese em Gel de Campo Pulsado , Genótipo , Humanos , Incidência , Epidemiologia Molecular , Prevalência , Técnica de Amplificação ao Acaso de DNA Polimórfico , Taiwan/epidemiologiaRESUMO
A field survey was conducted to investigate the presence of Borrelia burgdorferi sensu lato (s.l.) in six counties of Taiwan. Spirochetes were successfully isolated from one rodent ear sample out of 485 rodent ears and 53 live, fed tick (Ixodes granulatus) samples. The spirochetes were confirmed to be B. burgdorferi s.l. by real-time PCR. In addition, 23 of 113 tick samples were tested positive for Borrelia DNA according to real-time PCR. The Borrelia isolate from the rodent and the 23 Borrelia DNA samples from the ticks were identified as B. valaisiana-related genospecies by phylogenetic analysis based on flagellin gene sequences. These findings suggest that the Borrelia valaisiana-related strains are maintained in a zoonotic cycle between tick vectors and reservoir hosts in Taiwan.
Assuntos
Grupo Borrelia Burgdorferi/classificação , Grupo Borrelia Burgdorferi/isolamento & purificação , Ixodes/microbiologia , Roedores/microbiologia , Animais , Grupo Borrelia Burgdorferi/genética , Análise por Conglomerados , DNA Bacteriano/química , DNA Bacteriano/genética , Flagelina/genética , Dados de Sequência Molecular , Filogenia , Reação em Cadeia da Polimerase , Análise de Sequência de DNA , Homologia de Sequência , TaiwanRESUMO
BACKGROUND: High levels of Hepatoma Up-Regulated Protein (HURP) and Tousled-Like Kinase (TLK) transcripts are found in hepatocellular carcinoma. HURP overexpression induces anchorage-independent growth of 293-T cells and enhances a rough-eye phenotype resulting from tlk overexpression in Drosophila. In addition, both HURP and Mars, a Drosophila HURP sequence homologue, promote polymerization of mitotic spindles. Thus, the genetic interaction of mars with tlk might be required for accurate chromosome segregation. METHODS: To reveal whether chromosome fidelity was decreased, the frequency of gynandromorphy, an individual with both male and female characteristics, and of non-disjunction were measured in the progeny from parents with reduced mars and/or tlk activities and analyzed by Student's t-test. To show that the genetic interaction between mars and tlk is epistatic or parallel, a cytological analysis of embryos with either reduced or increased activities of mars and/or tlk was used to reveal defects in mitotic-spindle morphology and chromosome segregation. RESULTS: A significant but small fraction of the progeny from parents with reduced mars activity showed gynandromorphy and non-disjunction. Results of cytological analysis revealed that the decrease in chromosome fidelity was a result of delayed polymerization of the mitotic spindle, which led to asynchronous chromosome segregation in embryos that had reduced mars activity. By removing one copy of tousled-like kinase (tlk) from flies with reduced mars activity, chromosome fidelity was further reduced. This was indicated by an increased in the non-disjunction rate and more severe asynchrony. However, the morphology of the mitotic spindles in the embryos at metaphase where both gene activities were reduced was similar to that in mars embryos. Furthermore, tlk overexpression did not affect the morphology of the mitotic spindles and the cellular localization of Mars protein. CONCLUSION: Chromosome fidelity in progeny from parents with reduced mars and/or tlk activity was impaired. The results from cytological studies revealed that mars and tlk function in parallel and that a balance between mars activity and tlk activity is required for cells to progress through mitosis correctly, thus ensuring chromosome fidelity.
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Cromossomos/ultraestrutura , Proteínas de Drosophila/fisiologia , Drosophila melanogaster/metabolismo , Regulação da Expressão Gênica , Proteínas do Tecido Nervoso/fisiologia , Proteínas Serina-Treonina Quinases/fisiologia , Sequência de Aminoácidos , Animais , Ciclo Celular , Segregação de Cromossomos , Proteínas de Drosophila/biossíntese , Drosophila melanogaster/genética , Epistasia Genética , Humanos , Modelos Biológicos , Dados de Sequência Molecular , Proteínas do Tecido Nervoso/biossíntese , Proteínas Serina-Treonina Quinases/biossíntese , Estrutura Terciária de Proteína , Proteínas Associadas SAP90-PSD95 , Homologia de Sequência de Aminoácidos , Fuso AcromáticoRESUMO
The Torso (Tor) signaling pathway activates tailless (tll) expression by relieving tll repression. None of the repressors identified so far, such as Capicuo, Groucho and Tramtrack69 (Ttk69), bind to the tor response element (tor-RE) or fully elucidate tll repression. In this study, an expanded tll expression pattern was shown in embryos with reduced heat shock factor (hsf) and Trithorax-like (Trl) activities. The GAGA factor, GAF encoded by Trl, bound weakly to the tor-RE, and this binding was enhanced by both Hsf and Ttk69. A similar extent of expansion of tll expression was observed in embryos with simultaneous knockdown of hsf, Trl and ttk69 activities, and in embryos with constitutively active Tor. Hsf is a substrate of mitogen-activated protein kinase and S378 is the major phosphorylation site. Phosphorylation converts Hsf from a repressor to an activator that works with GAF to activate tll expression. In conclusion, the GAF/Hsf/Ttk69 complex binding to the tor-RE remodels local chromatin structure to repress tll expression and the Tor signaling pathway activate tll expression by modulating a dual transcriptional switch.
Assuntos
Proteínas de Ligação a DNA/genética , Proteínas de Drosophila/genética , Proteínas de Drosophila/metabolismo , Drosophila/genética , Regulação da Expressão Gênica , Receptores Proteína Tirosina Quinases/metabolismo , Proteínas Repressoras/genética , Fatores de Transcrição/metabolismo , Animais , Pegada de DNA , Proteínas de Ligação a DNA/antagonistas & inibidores , Proteínas de Ligação a DNA/química , Proteínas de Ligação a DNA/metabolismo , Drosophila/embriologia , Drosophila/metabolismo , Proteínas de Drosophila/antagonistas & inibidores , Proteínas de Drosophila/química , Embrião não Mamífero/metabolismo , Fatores de Transcrição de Choque Térmico , Histona Desacetilase 1 , Histona Desacetilases/metabolismo , Sistema de Sinalização das MAP Quinases , Proteínas Quinases Ativadas por Mitógeno/metabolismo , Interferência de RNA , Proteínas Repressoras/antagonistas & inibidores , Proteínas Repressoras/metabolismo , Elementos de Resposta , Serina/metabolismo , Fatores de Transcrição/antagonistas & inibidores , Fatores de Transcrição/química , Fatores de Transcrição/genética , Ativação TranscricionalRESUMO
In Taiwan, pertussis is a notifiable disease with a low incidence in recent years, and antimicrobial susceptibility testing for the causative agent, Bordetella pertussis, has not been reported to date. In May 2007, the Centers for Disease Control, Taiwan, was informed of a 1-month-old pertussis patient who did not respond to erythromycin treatment. In this study, we report the result of antimicrobial susceptibility testing performed for the suspected erythromycin-resistant isolate, as well as for an additional 27 B. pertussis clinical isolates that represented almost all epidemiologically unrelated isolates obtained throughout Taiwan between 2003 and 2007. All isolates were fully susceptible to azithromycin, erythromycin, clarithromycin and trimethoprim/sulfamethoxazole (MIC < or =0.047 mug ml(-1)). This result demonstrates the general susceptibility of B. pertussis to antimicrobial agents in vitro in Taiwan.
Assuntos
Antibacterianos/farmacologia , Bordetella pertussis/efeitos dos fármacos , Farmacorresistência Bacteriana , Eritromicina/farmacologia , Doenças do Prematuro , Coqueluche , Adolescente , Antibacterianos/uso terapêutico , Bordetella pertussis/genética , Criança , Pré-Escolar , Eletroforese em Gel de Campo Pulsado , Feminino , Humanos , Lactente , Recém-Nascido de Baixo Peso , Recém-Nascido , Recém-Nascido Prematuro , Doenças do Prematuro/tratamento farmacológico , Doenças do Prematuro/epidemiologia , Doenças do Prematuro/microbiologia , Masculino , Testes de Sensibilidade Microbiana , Taiwan , Coqueluche/tratamento farmacológico , Coqueluche/epidemiologia , Coqueluche/microbiologiaRESUMO
BACKGROUND: There has been a marked increase in the incidence of, and concern regarding, human Campylobacter jejuni and C. coli infections worldwide during the last decade. As the highest infectious disease control apparatus in Taiwan, we aimed to describe the character of Campylobacter isolates from infected children, as well as basic information about the patients, from December 2003 to February 2005. METHODS: A total of 894 fecal specimens were collected by several clinics and hospitals from children who had diarrhea, followed by plating onto selective media. Drug susceptibility test of the isolates from these specimens were conducted by disc diffusion method and their serotypes were also studied using commercial antisera made in Japan. RESULTS: The isolation rate of Campylobacter during these 15 months was 6.8% and was higher in winter (11.1%) than in other seasons. C. jejuni was the most prevalent (95.1%) species in northern Taiwan, comparable to other developed countries. Among the 61 Campylobacter isolates, most were resistant to tetracycline (93.4%), nalidixic acid (91.8%), ciprofloxacin (90.2%), and ampicillin (85.5%). Erythromycin-resistant isolates represented 3.3% of all isolates, suggesting that this drug may be the first choice for treatment. The serotypes of the 61 isolates were demonstrated and only 41.4% were typable. CONCLUSION: In this study, the Taiwan CDC provided an epidemiological analysis of Campylobacter infection, including the isolation rate, age, seasonal distribution, antimicrobial drug susceptibility patterns, and serotypes of the isolates from pediatric patients in northern Taiwan from 2003 to 2005.