RESUMO
The purpose of this study was to develop a small-group-focused suicide prevention program for elders with early-stage dementia and to assess its effects. This was a quasi-experimental study with a control group pretest-posttest design. A total of 62 elders diagnosed with early-stage dementia who were receiving care services at nine daycare centers in J City Korea participated in this study. The experimental group participated in the suicide prevention program twice a week for 5 weeks with a pretest and two posttests The developed suicide prevention program had a significant effect on the perceived health status, social support, depression, and suicidal ideation of elders with early-stage dementia. Nurses should integrate risk factors such as depression and protective factors such as health status and social support into a suicide prevention program. This community-based program in geriatric nursing practice can be effective in preventing suicide among elders with early-stage dementia.
Assuntos
Demência/psicologia , Avaliação de Programas e Projetos de Saúde , Características de Residência , Prevenção do Suicídio , Centros-Dia de Assistência à Saúde para Adultos/organização & administração , Idoso , Idoso de 80 Anos ou mais , Depressão/epidemiologia , Feminino , Enfermagem Geriátrica/métodos , Nível de Saúde , Humanos , Masculino , Desenvolvimento de Programas , República da Coreia/epidemiologia , Fatores de Risco , Apoio SocialRESUMO
BACKGROUND: Though the World Health Organization declared the 'elimination of leprosy as public health problem' in 2000, the disease remains endemic in many countries. Current trends in incidence of infection and disease are unclear. METHODS: Data on leprosy prevalence between 1977-2013 and data on new leprosy cases detected in the Republic of Korea between 1989-2013 were analysed by age, sex, clinical types, mode of detection, family history, disability grading and geographical distribution. RESULTS: Both prevalence and incidence have declined greatly. There has been a shift to an increased proportion of multibacillary disease, and older age groups, consistent with a dramatic decrease in infection transmission in recent decades. An increase in proportion of cases with family history of disease is consistent with these declines. There is evidence that declines in infection and disease have been greater in the north of the country, as revealed in patterns by place of birth over time. Cases in immigrants now form a substantial proportion of leprosy disease in the Republic of Korea. CONCLUSIONS: Leprosy has declined dramatically in the Republic of Korea in recent decades, and transmission of M. leprae may have effectively stopped. There remains a burden of care for individuals whose disease developed in the past, and there may be some additional newly detected cases among immigrants and among older individuals who acquired autochthonous infections decades ago.
Assuntos
Hanseníase/epidemiologia , Adulto , Idoso , Feminino , História do Século XX , História do Século XXI , Humanos , Incidência , Hanseníase/história , Hanseníase/microbiologia , Masculino , Pessoa de Meia-Idade , Mycobacterium leprae/genética , Mycobacterium leprae/isolamento & purificação , Mycobacterium leprae/fisiologia , República da Coreia/epidemiologia , Adulto JovemRESUMO
Glutamate is a commercially important chemical. It is used as a flavor enhancer and is a major raw material for producing industrially useful chemicals. A coryneform bacterium, Corynebacterium glutamicum, was isolated in 1956 by Japanese researchers as a glutamate-overproducing bacterium and since then, remarkable progress in glutamate production has been made using this microorganism. Currently, the global market for glutamate is over 2.5 million tons per year. Glutamate overproduction by C. glutamicum is induced by specific treatments-biotin limitation, addition of fatty acid ester surfactants such as Tween 40, and addition of ß-lactam antibiotics such as penicillin. Molecular biology and metabolic engineering studies on glutamate overproduction have revealed that metabolic flow is significantly altered by these treatments. These studies have also provided insight into the molecular mechanisms underlying these changes. In this chapter, we review our current understanding of the molecular mechanisms of glutamate overproduction in C. glutamicum, and we discuss the advances made by metabolic engineering of this microorganism.
Assuntos
Corynebacterium glutamicum/metabolismo , Ácido Glutâmico/biossíntese , Engenharia Metabólica , Ciclo do Ácido Cítrico/genética , Corynebacterium glutamicum/genética , Ácidos Cetoglutáricos/metabolismo , Redes e Vias Metabólicas/genética , Metilmalonil-CoA Descarboxilase/genética , Metilmalonil-CoA Descarboxilase/metabolismo , Subunidades ProteicasRESUMO
Glutamate overproduction by Corynebacterium glutamicum is triggered by treatment with penicillin or Tween 40 and is accompanied by a decrease in 2-oxoglutarate dehydrogenase complex (ODHC) activity. We have reported that de novo synthesis of OdhI, which inhibits ODHC activity by interacting specifically with the E1o subunit of ODHC (OdhA), is induced by penicillin, and that odhI overexpression induces glutamate overproduction in the absence of any triggers for glutamate overproduction. In this study, to determine the function of OdhI in glutamate overproduction by C. glutamicum, changes in OdhI levels and phosphorylation status during penicillin- and Tween 40-induced glutamate overproduction were examined by western blot. The synthesis of both unphosphorylated and phosphorylated OdhI was increased by addition of Tween 40 or penicillin and the levels of unphosphorylated OdhI, which can inhibit ODHC activity, was significantly higher than those of phosphorylated OdhI, which is unable to inhibit ODHC activity. Meanwhile, the OdhA levels were maintained throughout the culture. These results indicate that OdhI synthesis is induced by additions of penicillin and Tween 40 and most synthesized OdhI is unphosphorylated, resulting in the decrease in ODHC activity and glutamate overproduction. Similarly, in the odhI-overexpressing strain, both unphosphorylated and phosphorylated OdhI were synthesized, while the levels of OdhA were nearly constant throughout culture. Our results suggest that high level of unphosphorylated OdhI regulates glutamate overproduction by C. glutamicum.
Assuntos
Proteínas de Bactérias/metabolismo , Corynebacterium glutamicum/metabolismo , Ácido Glutâmico/metabolismo , Penicilinas/farmacologia , Polissorbatos/farmacologia , Proteínas de Bactérias/genética , Corynebacterium glutamicum/efeitos dos fármacos , Fosforilação/efeitos dos fármacosRESUMO
To end the decade-long, obstinately stagnant number of new leprosy cases, there is an urgent need for field-applicable diagnostic tools that detect infection with Mycobacterium leprae, leprosy's etiologic agent. Since immunity against M. leprae is characterized by humoral and cellular markers, we developed a lateral flow test measuring multiple host proteins based on six previously identified biomarkers for various leprosy phenotypes. This multi-biomarker test (MBT) demonstrated feasibility of quantitative detection of six host serum proteins simultaneously, jointly allowing discrimination of patients with multibacillary and paucibacillary leprosy from control individuals in high and low leprosy endemic areas. Pilot testing of fingerstick blood showed similar MBT performance in point-of-care (POC) settings as observed for plasma and serum. Thus, this newly developed prototype MBT measures six biomarkers covering immunity against M. leprae across the leprosy spectrum. The MBT thereby provides the basis for immunodiagnostic POC tests for leprosy with potential for other (infectious) diseases as well.
RESUMO
We found that penicillin-induced glutamate production by Corynebacterium glutamicum is inhibited when a de novo protein synthesis inhibitor, chloramphenicol, is added simultaneously with penicillin. When chloramphenicol was added 4 h after penicillin addition, glutamate production was essentially unaffected. (3)H-Leucine incorporation experiments revealed that protein synthesis continued for 1 h after penicillin addition and then gradually decreased. These results suggest that de novo protein synthesis within 4 h of penicillin treatment is required for the induction of glutamate production. To identify the protein(s) necessary for penicillin-induced glutamate production, proteome analysis of penicillin-treated C. glutamicum cells was performed with two-dimensional gel electrophoresis. Of more than 500 proteins detected, the amount of 13 proteins, including OdhI (an inhibitory protein for 2-oxoglutarate dehydrogenase complex), significantly increased upon penicillin treatment. Artificial overexpression of the odhI gene resulted in the decreased specific activity of the 2-oxoglutarate dehydrogenase complex and increased glutamate production without any triggers. These results suggest that the de novo synthesis of OdhI is the necessary factor for penicillin-induced glutamate overproduction by C. glutamicum. Moreover, continuous glutamate production was achieved by overexpression of odhI without any triggers. Thus, the odhI-overexpressing strain of C. glutamicum can be useful for efficient glutamate production.
Assuntos
Proteínas de Bactérias/biossíntese , Corynebacterium glutamicum/enzimologia , Ácido Glutâmico/biossíntese , Penicilinas/metabolismo , Ativação Transcricional , Antibacterianos/farmacologia , Cloranfenicol/farmacologia , Eletroforese em Gel Bidimensional , Expressão Gênica , Proteoma/análise , Coloração e Rotulagem/métodos , Trítio/metabolismoRESUMO
Recent studies have suggested that a decrease in the specific activity of the 2-oxoglutarate dehydrogenase complex (ODHC) is important for glutamate overproduction by Corynebacterium glutamicum. To further investigate the role of the odhA gene and its product in this process, we constructed the recombinant strains of C. glutamicum in which the expression of the odhA and its product could be controlled by odhA overexpression and odhA antisense RNA expression. We examined changes in glutamate production and ODHC specific activity of the constructed strains during glutamate production triggered by Tween 40 addition. The ODHC specific activity increased with odhA overexpression, resulting in dramatically reduced glutamate production despite Tween 40 addition, indicating that a decrease in the specific activity of ODHC is required for glutamate production induced by Tween 40 addition. However, odhA antisense RNA expression alone did not result in glutamate overproduction in spite of the decrease in ODHC specific activity. Rather, it enhanced glutamate production triggered by Tween 40 addition due to the additional decrease in ODHC specific activity, suggesting that odhA antisense RNA expression is effective in enhancing Tween-40-triggered glutamate overproduction. Our results suggest that a change in ODHC specific activity is critical but is not the only factor responsible for glutamate overproduction by C. glutamicum.
Assuntos
Proteínas de Bactérias/biossíntese , Corynebacterium glutamicum/metabolismo , Ácido Glutâmico/metabolismo , Complexo Cetoglutarato Desidrogenase/biossíntese , RNA Antissenso/genética , Proteínas de Bactérias/genética , Dosagem de Genes , Inativação Gênica , Complexo Cetoglutarato Desidrogenase/genéticaRESUMO
Drug resistance in Mycobacterium leprae is a significant problem in countries where leprosy is endemic. A sensitive, specific, and high-throughput reverse blot hybridization assay (REBA) for the detection of genotypic resistance to rifampicin (RIF) was designed and evaluated. It has been shown that resistance to RIF in M. leprae involves mutations in the rpoB gene encoding the -subunit of the RNA polymerase. The PCR-REBA simultaneously detects both 6 wild-type regions and 5 different mutations (507 AGC, 513 GTG, 516 TAT, 531 ATG, and 531 TTC) including the most prevalent mutations at positions 507 and 531. Thirty-one clinical isolates provided by Korea Institute of Hansen-s Disease were analyzed by PCR-REBA with RIF resistance of rpoB gene. As a result, missense mutations at codons 507 AGC and 531 ATG with 2-nucleotide substitutions were found in one sample, and a missense mutation at codon 516 TAT and ΔWT6 (deletion of 530-534) was found in another sample. These cases were confirmed by DNA sequence analysis. This rapid, simple, and highly sensitive assay provides a practical alternative to sequencing for genotypic evaluation of RIF resistance in M. leprae.
Assuntos
Antibióticos Antituberculose/farmacologia , Mycobacterium leprae/efeitos dos fármacos , Mycobacterium leprae/genética , Hibridização de Ácido Nucleico/métodos , Reação em Cadeia da Polimerase/métodos , Rifampina/farmacologia , Sequência de Aminoácidos , Sequência de Bases , RNA Polimerases Dirigidas por DNA/genética , Farmacorresistência Bacteriana , Humanos , Dados de Sequência Molecular , Mutação , Mycobacterium leprae/isolamento & purificação , Sensibilidade e EspecificidadeRESUMO
DNA-PCR and reverse transcription (RT)-PCR for the 18-kDa protein of Mycobacterium leprae were used to examine the efficacy of multi-drug therapy (MDT) in leprosy. MDT was administered for 0-24 months. Fourteen (63.6%) of 22 patients showed positive PCR results after treatment for 12 months and the positive results decreased to 30% after 24 months of MDT. These results did not correlate with the bacterial index (BI) or the IgM antibody titre for the phenolic glycolipid (PGL)-1. One-dimensional densitometric analysis of agarose gels from PCR from the longitudinal study showed a gradual reduction of the 360-bp band after 12-24 months of MDT. RT-PCR for mRNA of the 18-kDa protein successfully tracked bacterial RNA changes in the biopsies and confirmed a decrease in the RNA of M. leprae in patients after MDT for 12 months. Thus, DNA- and RT-PCR for the 18-kDa protein of M. leprae are effective in assessing the efficacy of MDT for leprosy.
Assuntos
Antígenos de Bactérias , Proteínas de Bactérias/genética , Hansenostáticos/uso terapêutico , Hanseníase/microbiologia , Mycobacterium leprae/genética , Reação em Cadeia da Polimerase/métodos , Animais , Biópsia , DNA Bacteriano/análise , Densitometria , Quimioterapia Combinada , Eletroforese em Gel de Ágar , Glicolipídeos/genética , Glicolipídeos/imunologia , Humanos , Imunoglobulina M/sangue , Hanseníase/tratamento farmacológico , Estudos Longitudinais , Camundongos , Camundongos Nus , Mycobacterium leprae/efeitos dos fármacos , Mycobacterium leprae/imunologia , RNA Mensageiro/análise , Reprodutibilidade dos Testes , Reação em Cadeia da Polimerase Via Transcriptase ReversaRESUMO
Co-culture of human skeletal muscle myoblasts and fibroblasts was performed on a dendrimer-immobilized surface. Morphological changes by cytoskeletal variation were observed, compared with culture on a conventional tissue culture surface, and preferential growth of myoblasts can be achieved by inhibition of fibroblast growth.
Assuntos
Dendrímeros/metabolismo , Fibroblastos/citologia , Fibroblastos/metabolismo , Mioblastos Esqueléticos/citologia , Mioblastos Esqueléticos/metabolismo , Adesão Celular , Proliferação de Células , Forma Celular , Células Cultivadas , Técnicas de Cocultura , Citoesqueleto/metabolismo , Dendrímeros/química , Humanos , Músculo Esquelético/citologiaRESUMO
Leprosy is an infectious, neurodegenerative disease of humans caused by Mycobacterium leprae. Despite effective control programs, the incidence of leprosy remains stubbornly high, suggesting that transmission may be more common than expected. The rationale of this work was to use bioinformatics and comparative genomics to identify potentially antigenic proteins for diagnostic purposes. This approach defined three classes of proteins: those restricted to M. leprae (class I), those present in M. leprae with orthologues in other organisms besides mycobacteria (class II), and exported or surface-exposed proteins (class III). Twelve genes (two class I, four class II, and six class III proteins) were cloned in Escherichia coli, and their protein products were purified. Six of these proteins were detected in cell extracts of M. leprae by immunoblotting. The immunogenicity of each recombinant protein was then investigated in leprosy patients by measuring the reactivity of circulating antibody and gamma interferon (IFN-gamma) responses in T-cell restimulation assays. Several class II and class III proteins were recognized by circulating antibodies. Importantly, most class II proteins elicited IFN-gamma responses that were significantly stronger than those produced by previously identified antigens. Among them, two class II proteins, ML0308 and ML2498, showed marked humoral and cellular immunogenicity, therefore providing promising candidates for the diagnosis of both tuberculoid and lepromatous forms of leprosy.