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1.
J Microbiol Immunol Infect ; 56(6): 1236-1244, 2023 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-37690869

RESUMO

BACKGROUND: Pyrazinamide (PZA) and fluoroquinolone (FQ), particularly moxifloxacin (MXF), are essential drugs in the World Health Organization (WHO) recommended short-course regimen to treat drug-susceptible tuberculosis (TB). METHODS: To understand the extent of PZA and MXF susceptibility in general TB cases in Taiwan, we conducted retrospective analyses of 385 conservative Mycobacterium tuberculosis complex (MTBC) isolates identified from 4 TB laboratories in different regions of Taiwan. The case information was obtained from the TB registry. Genotypic drug susceptibility testing (DST) was performed by sequencing drug-resistance associated genes, PZA (pncA) and FQ (gyrA, and gyrB). Phenotypic DST was determined using the Bactec MGIT 960 system or the agar proportion method. Genotyping was carried out using spacer oligonucleotide typing. RESULTS: In this study, 4.7% (18/385) cases' isolates harbored pncA mutations and 7.0% (27/385) cases' isolates harbored gyrA or gyrB mutation. Notably, pncA mutation was associated with Beijing family genotypes (P = 0.028), East African-Indian (EAI) genotypes (P = 0.047) and MDR-TB (P < 0.001). Whereas, gyrA or gyrB mutation was associated with EAI genotypes (P = 0.020) and MDR-TB (P = 0.006). In addition, a statistically significant difference was found between the favorable outcomes using active and inactive PZA (P = 0.009) in 38 case isolates with any pncA, gyrA, or gyrB mutation. CONCLUSION: We concluded that routine PZA and FQ susceptibility tests are recommended for guiding the treatment of TB.


Assuntos
Mycobacterium tuberculosis , Tuberculose Resistente a Múltiplos Medicamentos , Tuberculose , Humanos , Pirazinamida/farmacologia , Pirazinamida/uso terapêutico , Antituberculosos/farmacologia , Antituberculosos/uso terapêutico , Fluoroquinolonas/farmacologia , Fluoroquinolonas/uso terapêutico , Testes de Sensibilidade Microbiana , Taiwan , Estudos Retrospectivos , Farmacorresistência Bacteriana Múltipla , Amidoidrolases/genética , Tuberculose/tratamento farmacológico , Tuberculose/microbiologia , Tuberculose Resistente a Múltiplos Medicamentos/tratamento farmacológico , Tuberculose Resistente a Múltiplos Medicamentos/epidemiologia , Tuberculose Resistente a Múltiplos Medicamentos/microbiologia , Mutação , Moxifloxacina/farmacologia , Moxifloxacina/uso terapêutico
2.
Front Pharmacol ; 13: 853818, 2022.
Artigo em Inglês | MEDLINE | ID: mdl-35370629

RESUMO

Septicemia is a severe inflammatory response caused by the invasion of foreign pathogens. Severe sepsis-induced shock and multiple organ failure are the two main causes of patient death. The overexpression of many proinflammatory cytokines, such as TNF-α, IL-1ß, and IL-6, is closely related to severe sepsis. Although the treatment of sepsis has been subject to many major breakthroughs of late, the treatment of patients with septic shock is still accompanied by a high mortality rate. In our previous research, we used computer simulations to design the multifunctional peptide KCF18 that can bind to TNF-α, IL-1ß, and IL-6 based on the binding regions of receptors and proinflammatory cytokines. In this study, proinflammatory cytokines were used to stimulate human monocytes to trigger an inflammatory response, and the anti-inflammatory ability of the multifunctional KCF18 peptide was further investigated. Cell experiments demonstrated that KCF18 significantly reduced the binding of proinflammatory cytokines to their cognate receptors and inhibited the mRNA and protein expressions of TNF-α, IL-1ß, and IL-6. It could also reduce the expression of reactive oxygen species induced by cytokines in human monocytes. KCF18 could effectively decrease the p65 nucleus translocation induced by cytokines, and a mice endotoxemia experiment demonstrated that KCF18 could reduce the expression of IL-6 and the increase of white blood cells in the blood stimulated by lipopolysaccharides. According to our study of tissue sections, KCF18 alleviated liver inflammation. By reducing the release of cytokines in plasma and directly affecting vascular cells, KCF18 is believed to significantly reduce the risk of vascular inflammation.

3.
Sci Rep ; 11(1): 644, 2021 01 12.
Artigo em Inglês | MEDLINE | ID: mdl-33436772

RESUMO

Ephrin type-A receptor 10 (EPHA10) has been implicated as a potential target for breast and prostate cancer therapy. However, its involvement in oral squamous cell carcinoma (OSCC) remains unclear. We demonstrated that EPHA10 supports in vivo tumor growth and lymphatic metastasis of OSCC cells. OSCC cell migration, epithelial mesenchymal transition (EMT), and sphere formation were found to be regulated by EPHA10, and EPHA10 was found to drive expression of some EMT- and stemness-associated transcription factors. Among EPHA10 ligands, exogenous ephrin A4 (EFNA4) induced the most OSCC cell migration and sphere formation, as well as up-regulation of SNAIL, NANOG, and OCT4. These effects were abolished by extracellular signal-regulated kinase (ERK) inhibition and NANOG knockdown. Also, EPHA10 was required for EFNA4-induced cell migration, sphere formation, and expression of NANOG and OCT4 mRNA. Our microarray dataset revealed that EFNA4 mRNA expression was associated with expression of NANOG and OCT4 mRNA, and OSCC patients showing high co-expression of EFNA4 with NANOG or OCT4 mRNA demonstrated poor recurrence-free survival rates. Targeting forward signaling of the EFNA4-EPHA10 axis may be a promising therapeutic approach for oral malignancies, and the combination of EFNA4 mRNA and downstream gene expression may be a useful prognostic biomarker for OSCC.


Assuntos
Movimento Celular , Efrina-A4/metabolismo , Regulação Neoplásica da Expressão Gênica , Neoplasias Bucais/patologia , Proteína Homeobox Nanog/metabolismo , Receptores da Família Eph/metabolismo , Esferoides Celulares/patologia , Animais , Apoptose , Biomarcadores Tumorais/genética , Biomarcadores Tumorais/metabolismo , Carcinoma de Células Escamosas/genética , Carcinoma de Células Escamosas/metabolismo , Carcinoma de Células Escamosas/patologia , Proliferação de Células , Efrina-A4/genética , Transição Epitelial-Mesenquimal , Humanos , Masculino , Camundongos , Camundongos Endogâmicos BALB C , Camundongos Nus , Neoplasias Bucais/genética , Neoplasias Bucais/metabolismo , Proteína Homeobox Nanog/genética , Células-Tronco Neoplásicas/metabolismo , Células-Tronco Neoplásicas/patologia , Prognóstico , Receptores da Família Eph/genética , Esferoides Celulares/metabolismo , Taxa de Sobrevida , Células Tumorais Cultivadas , Ensaios Antitumorais Modelo de Xenoenxerto
4.
Immunology ; 160(1): 38-51, 2020 05.
Artigo em Inglês | MEDLINE | ID: mdl-32027025

RESUMO

First discovered on the natural killer (NK) cell, the cell surface inhibitory receptor sialic acid-binding immunoglobulin-like lectin-7 (Siglec-7) is known for regulating many important biological activities. However, the detail regulatory mechanism for Siglec-7 expression in NK cells currently remains unclear. In this study, we aimed to investigate how cell surface Siglec-7 expression is regulated and found that, in both NK cell lines and peripheral NK cells, transcription was the main regulatory step. Furthermore, when NK-92MI and peripheral NK cells were treated with DNA methyltransferase (DNMT) inhibitor, the CpG island, with 9 CpG sites, in 5' Siglec-7 promoter became noticeably hypomethylated, and Siglec-7 expression increased in both RNA transcript and surface protein. Within this CpG island, we identified both CpG 8 and CpG 9 as two key regulators responsible for Siglec-7 expression. Additionally, by using histone deacetylases (HDAC) inhibitor, butyric acid, we showed that Siglec-7 expression was also subjected to the histone modification. And a combined treatment with both 5-azacytidine and butyric acid showed an additive effect on Siglec-7 transcript expression in peripheral NK cells.


Assuntos
Antígenos de Diferenciação Mielomonocítica/genética , Epigênese Genética/imunologia , Células Matadoras Naturais/imunologia , Lectinas/genética , Antígenos de Diferenciação Mielomonocítica/metabolismo , Azacitidina/farmacologia , Ácido Butírico/farmacologia , Linhagem Celular , Ilhas de CpG/genética , Metilação de DNA/efeitos dos fármacos , Metilação de DNA/imunologia , Epigênese Genética/efeitos dos fármacos , Código das Histonas/efeitos dos fármacos , Inibidores de Histona Desacetilases/farmacologia , Histona Desacetilases/metabolismo , Humanos , Células Matadoras Naturais/efeitos dos fármacos , Células Matadoras Naturais/metabolismo , Lectinas/metabolismo , Regiões Promotoras Genéticas/genética , RNA-Seq , Transcrição Gênica/efeitos dos fármacos , Transcrição Gênica/imunologia
5.
Acta Crystallogr F Struct Biol Commun ; 76(Pt 1): 25-30, 2020 Jan 01.
Artigo em Inglês | MEDLINE | ID: mdl-31929183

RESUMO

RcnR is a transcription factor that regulates the homeostasis of cobalt and nickel in bacterial cells. Escherichia coli RcnR was crystallized with DNA that encompasses the DNA-binding site. X-ray diffraction data were collected to 2.9 Šresolution. The crystal belonged to space group P6122 or P6522, with unit-cell parameters a = b = 73.59, c = 157.66 Å, α = ß = 90, γ = 120°.


Assuntos
Cobalto/química , Proteínas de Escherichia coli/química , Níquel/química , Proteínas Repressoras/química , Cobalto/metabolismo , Cristalografia por Raios X , DNA , Escherichia coli/química , Escherichia coli/genética , Proteínas de Escherichia coli/isolamento & purificação , Modelos Moleculares , Níquel/metabolismo , Proteínas Repressoras/isolamento & purificação , Difração de Raios X
6.
Rehabil Nurs ; 45(4): 195-203, 2020.
Artigo em Inglês | MEDLINE | ID: mdl-29864046

RESUMO

PURPOSE: The aim of the study was to develop the Healthy Beat Acupunch (HBA) exercise program and evaluate its feasibility for community older adults. DESIGN: Stage I: The Delphi technique was used to consult 16 experts to develop the program. Stage II: A preexperimental, one-group, posttest-only design was used to pilot-test the program feasibility with 31 older adults. METHODS: After 4 weeks of interventions, participants evaluated the program based on four criteria (simplicity, safety, suitability, and helpfulness) and responded to five open-ended questions. FINDINGS: The developed HBA program from Stage I includes three phases with 24 motions and takes 40 minutes to complete. Program feasibility in Stage II showed average scores ranged from 8.84 ± 1.32 to 9.97 ± 0.18. CONCLUSION: Both experts and elderly participants confirmed that the HBA program was simple, safe, suitable, and helpful to older adults. CLINICAL RELEVANCE: The HBA program provides older adults with a new set of exercise options.


Assuntos
Terapia por Acupuntura/normas , Desenvolvimento de Programas/métodos , Terapia por Acupuntura/métodos , Terapia por Acupuntura/estatística & dados numéricos , Adulto , Estudos de Viabilidade , Feminino , Geriatria/métodos , Humanos , Masculino , Pessoa de Meia-Idade , Desenvolvimento de Programas/estatística & dados numéricos
7.
Immunobiology ; 224(5): 649-658, 2019 09.
Artigo em Inglês | MEDLINE | ID: mdl-31421859

RESUMO

Transforming growth factor-ß (TGF-ß) is known to function as a dual role regulatory cytokine for being either a suppresser or promoter during tumor initiation and progression. In solid tumors, TGF-ß secreted from tumor microenvironment acts as a suppresser against host immunity, like natural killer (NK) cells, to favor tumor evasion. However, besides solid tumors, the underlying mechanism of how TGF-ß regulates leukemogenesis, tumor progression, immunoediting, and NK function is still not clear in detail. In this study, we found that TGF-ß induced leukemia MEG-01 and U937 cells to become less sensitive to NK-92MI targeting by down-regulating CD48, a ligand for NK activating receptor 2B4, but not down-regulating other tumor-associated carbohydrate antigens (TACAs). In CD48-knockdown cells, cells responding to NK-92MI targeting displayed a phenotype of less NK susceptibility and cell conjugation. On the other hand, when NK cells were treated with TGF-ß, TGF-ß suppressed NK recognition, degranulation, and killing activity in time-dependent manner by regulating ICAM-1 binding capacity instead of affecting expressions of activating and inhibitory receptors. Taken together, both leukemia cells and immune NK cells could be regulated by TGF-ß through suppressing leukemia cell surface CD48 to escape from host surveillance and down-regulating NK cell surface ICAM-1 binding activity to impair NK functions, respectively. Our results suggested that TGF-ß had effect in leukemia similar to that observed in solid tumors but through different regulatory mechanism.


Assuntos
Antígeno CD48/metabolismo , Células Matadoras Naturais/imunologia , Células Matadoras Naturais/metabolismo , Leucemia/etiologia , Leucemia/metabolismo , Fator de Crescimento Transformador beta/metabolismo , Antígeno CD48/genética , Degranulação Celular , Linhagem Celular Tumoral , Citotoxicidade Imunológica/efeitos dos fármacos , Suscetibilidade a Doenças , Humanos , Vigilância Imunológica , Molécula 1 de Adesão Intercelular/imunologia , Molécula 1 de Adesão Intercelular/metabolismo , Leucemia/patologia , Proteína 1 de Membrana Associada ao Lisossomo/metabolismo , Receptores de Células Matadoras Naturais/metabolismo , Fator de Crescimento Transformador beta/farmacologia
8.
Inorg Chem ; 58(20): 13639-13653, 2019 Oct 21.
Artigo em Inglês | MEDLINE | ID: mdl-31247878

RESUMO

E. coli RcnR (resistance to cobalt and nickel regulator) is a homotetrameric DNA binding protein that regulates the expression of a Ni(II) and Co(II) exporter (RcnAB) by derepressing expression of rcnA and rcnB in response to binding Co(II) or Ni(II). Prior studies have shown that the cognate metal ions, Ni(II) and Co(II), bind in six-coordinate sites at subunit interfaces and are distinguished from noncognate metals (Cu(I), Cu(II), and Zn(II)) by coordination number and ligand selection. In analogy with FrmR, a formaldehyde-responsive transcriptional regulator in the RcnR/CsoR family, the interfacial site allows the metal ions to "cross-link" the N-terminal domain of one subunit with the invariant Cys35 residue in another, which has been deemed to be key to mediating the allosteric response of the tetrameric protein to metal binding. Through the use of mutagenesis to disconnect one subunit from the metal-mediated cross-link, X-ray absorption spectroscopy (XAS) as a structural probe, LacZ reporter assays, and metal binding studies using isothermal titration calorimetry (ITC), the work presented here shows that neither the interfacial binding site nor the coordination number of Ni(II) is important to the allosteric response to binding of this cognate metal ion. The opposite is found for the other cognate metal ion, Co(II), with respect to the interfacial binding site, suggesting that the molecular mechanisms for transcriptional regulation by the two ions are distinct. The metal binding studies reveal that tight metal binding is maintained in the variant. XAS is further used to demonstrate that His33 is not a ligand for Co(II), Ni(II), or Zn(II) in WT-RcnR. The results are discussed in the context of the overall understanding of the molecular mechanisms of metallosensors.


Assuntos
Proteínas de Escherichia coli/metabolismo , Níquel/metabolismo , Proteínas Repressoras/metabolismo , Proteínas de Escherichia coli/química , Proteínas de Escherichia coli/genética , Modelos Moleculares , Níquel/química , Proteínas Repressoras/química , Proteínas Repressoras/genética
9.
Sci Rep ; 9(1): 2317, 2019 02 19.
Artigo em Inglês | MEDLINE | ID: mdl-30783144

RESUMO

Chronic inflammation is a pivotal event in the pathogenesis of cardiovascular diseases, including atherosclerosis, restenosis, and coronary artery disease. The efficacy of current treatment or preventive strategies for such inflammation is still inadequate. Thus, new anti-inflammatory strategies are needed. In this study, based on molecular docking and structural analysis, a potential peptide KCF18 with amphiphilic properties (positively charged and hydrophobic residues) derived from the receptors of proinflammatory cytokines was designed to inhibit cytokine-induced inflammatory response. Simulations suggested that KCF18 could bind to cytokines simultaneously, and electrostatic interactions were dominant. Surface plasmon resonance detection showed that KCF18 bound to both tumor necrosis factor-α (TNF-α) and interleukin-6, which is consistent with MM/PBSA binding free energy calculations. The cell experiments showed that KCF18 significantly reduced the binding of proinflammatory cytokines to their cognate receptors, suppressed TNF-α mRNA expression and monocyte binding and transmigration, and alleviated the infiltration of white blood cells in a peritonitis mouse model. The designed peptide KCF18 could remarkably diminish the risk of vascular inflammation by decreasing plasma cytokines release and by directly acting on the vascular endothelium. This study demonstrated that a combination of structure-based in silico design calculations, together with experimental measurements can be used to develop potential anti-inflammatory agents.


Assuntos
Inflamação/tratamento farmacológico , Inflamação/metabolismo , Peptídeos/química , Peptídeos/uso terapêutico , Receptores de Citocinas/química , Humanos , Interleucina-1beta/metabolismo , Interleucina-1beta/farmacologia , Interleucina-6/metabolismo , Interleucina-6/farmacologia , Ligação Proteica , Células THP-1 , Fator de Necrose Tumoral alfa/metabolismo , Fator de Necrose Tumoral alfa/farmacologia
10.
Inorg Chem ; 57(20): 12521-12535, 2018 Oct 15.
Artigo em Inglês | MEDLINE | ID: mdl-30281299

RESUMO

Superoxide dismutases (SODs) utilize a ping-pong mechanism in which a redox-active metal cycles between oxidized and reduced forms that differ by one electron to catalyze the disproportionation of superoxide to dioxygen and hydrogen peroxide. Nickel-dependent SOD (NiSOD) is a unique biological solution for controlling superoxide levels. This enzyme relies on the use of cysteinate ligands to bring the Ni(III/II) redox couple into the range required for catalysis (∼300 mV vs. NHE). The use of cysteine thiolates, which are not found in any other SOD, is a curious choice because of their well-known oxidation by peroxide and dioxygen. The NiSOD active site cysteinate ligands are resistant to oxidation, and prior studies of synthetic and computational models point to the backbone N-donors in the active site (the N-terminal amine and the amide N atom of Cys2) as being involved in stabilizing the cysteines to oxidation. To test the role of the backbone N-donors, we have constructed a variant of NiSOD wherein an alanine residue was added to the N-terminus (Ala0-NiSOD), effectively altering the amine ligand to an amide. X-ray absorption, electronic absorption, and magnetic circular dichroism (MCD) spectroscopic analyses of as-isolated Ala0-NiSOD coupled with density functional theory (DFT) geometry optimized models that were evaluated on the basis of the spectroscopic data within the framework of DFT and time-dependent DFT computations are consistent with a diamagnetic Ni(II) site with two cysteinate, one His1 amide, and one Cys2 amidate ligands. The variant protein is catalytically inactive, has an altered electronic absorption spectrum associated with the nickel site, and is sensitive to oxidation. Mass spectrometric analysis of the protein exposed to air shows the presence of a mixture of oxidation products, the principal ones being a disulfide, a bis-sulfenate, and a bis-sulfinate derived from the active site cysteine ligands. Details of the electronic structure of the Ni(III) site available from the DFT calculations point to subtle changes in the unpaired spin density on the S-donors as being responsible for the altered sensitivity of Ala0-NiSOD to O2.


Assuntos
Amidas/metabolismo , Aminas/metabolismo , Níquel/química , Superóxido Dismutase/metabolismo , Amidas/química , Aminas/química , Escherichia coli/metabolismo , Regulação Enzimológica da Expressão Gênica , Modelos Moleculares , Conformação Proteica , Superóxido Dismutase/química
11.
Inorg Chem ; 57(20): 12588-12595, 2018 Oct 15.
Artigo em Inglês | MEDLINE | ID: mdl-30252455

RESUMO

α-Ketoglutarate (αKG) dependent oxygenases comprise a large superfamily of enzymes that activate O2 for varied reactions. While most of these enzymes contain a nonheme Fe bound by a His2(Asp/Glu) facial triad, a small number of αKG-dependent halogenases require only the two His ligands to bind Fe and activate O2. The enzyme "factor inhibiting HIF" (FIH) contains a His2Asp facial triad and selectively hydroxylates polypeptides; however, removal of the Asp ligand in the Asp201→Gly variant leads to a highly active enzyme, seemingly without a complete facial triad. Herein, we report on the formation of an Fe-Cl cofactor structure for the Asp201→Gly FIH variant using X-ray absorption spectroscopy (XAS), which provides insight into the structure of the His2Cl facial triad found in halogenases. The Asp201→Gly variant supports anion dependent peptide hydroxylation, demonstrating the requirement for a complete His2X facial triad to support O2 reactivity. Our results indicated that exogenous ligand binding to form a complete His2X facial triad was essential for O2 activation and provides a structural model for the His2Cl-bound nonheme Fe found in halogenases.


Assuntos
Cloretos/química , Ferro/metabolismo , Oxigenases de Função Mista/metabolismo , Oxigênio/metabolismo , Proteínas Repressoras/metabolismo , Absorciometria de Fóton , Substituição de Aminoácidos , Cloretos/metabolismo , Ferro/química , Ligantes , Oxigenases de Função Mista/química , Ligação Proteica , Conformação Proteica , Proteínas Repressoras/química
12.
Biochemistry ; 57(20): 2932-2942, 2018 05 22.
Artigo em Inglês | MEDLINE | ID: mdl-29708738

RESUMO

Helicobacter pylori is a human pathogen that colonizes the stomach, is the major cause of ulcers, and has been associated with stomach cancers. To survive in the acidic environment of the stomach, H. pylori uses urease, a nickel-dependent enzyme, to produce ammonia for maintenance of cellular pH. The bacteria produce apo-urease in large quantities and activate it by incorporating nickel under acid shock conditions. Urease nickel incorporation requires the urease-specific metallochaperone UreE and the (UreFGH)2 maturation complex. In addition, the H. pylori nickel urease maturation pathway recruits accessory proteins from the [NiFe] hydrogenase maturation pathway, namely, HypA and HypB. HypA and UreE dimers (UreE2) are known to form a protein complex, the role of which in urease maturation is largely unknown. Herein, we examine the nickel-binding properties and protein-protein interactions of HypA and UreE2 using isothermal titration calorimetry and fluorometric methods under neutral and acidic pH conditions to gain insight into the roles played by HypA in urease maturation. The results reveal that HypA and UreE2 form a stable complex with micromolar affinity that protects UreE from hydrolytic degradation. The HypA·UreE2 complex contains a unique high-affinity (nanomolar) Ni2+-binding site that is maintained under conditions designed to mimic acid shock (pH 6.3). The data are interpreted in terms of a proposed mechanism wherein HypA and UreE2 act as co-metallochaperones that target the delivery of Ni2+ to apo-urease with high fidelity.


Assuntos
Proteínas de Bactérias/química , Proteínas de Transporte/química , Helicobacter pylori/química , Complexos Multiproteicos/química , Proteínas de Bactérias/genética , Sítios de Ligação , Proteínas de Transporte/genética , Helicobacter pylori/genética , Helicobacter pylori/patogenicidade , Humanos , Metalochaperonas/química , Metalochaperonas/genética , Complexos Multiproteicos/genética , Níquel/química , Ligação Proteica , Domínios Proteicos
13.
Am J Geriatr Psychiatry ; 26(5): 511-520, 2018 05.
Artigo em Inglês | MEDLINE | ID: mdl-29433845

RESUMO

OBJECTIVES: To test the long-term effects of the 12-month Healthy Beat Acupunch (HBA) exercise program on the self-perceived health and sleep quality of older adults in community care centers, and to compare the effects of two delivery methods: instructor-led HBA for the first 6 months and DVD-guided HBA for another 6 months. DESIGN: Cluster-randomized controlled trial. SETTING: Eight community care centers. PARTICIPANTS: In total, 232 participants were recruited from eight community care centers, and cluster-randomized to the experimental (4 centers, N = 113) and control (4 centers, N = 119) groups. INTERVENTION: The experimental group received the instructor-led HBA program 3 times weekly for the first 6 months, followed by the DVD-guided HBA program for another 6 months. MEASUREMENTS: Self-perceived health and sleep quality were assessed using the Short Form Health Survey and the Pittsburgh Sleep Quality Index, respectively, at baseline and every 3 months for 1 year. RESULTS: The experimental group reported more favorable self-perceived physical and mental health, higher subjective sleep quality, and less daytime dysfunction than did the control group. Effect sizes of physical health and sleep quality increased from the instructor-led stage to the DVD-guided stage; the effect size of physical health showed the most significant change, increasing from 0.38 in the instructor-led stage to 0.55 in the DVD-guided stage. CONCLUSIONS: The exercise program consisting of the instructor-led class, followed by the DVD-guided class, was an effective and feasible longitudinal program for older adults in community care centers.


Assuntos
Terapia por Exercício/métodos , Exercício Físico , Nível de Saúde , Sono , Idoso , Idoso de 80 Anos ou mais , Autoavaliação Diagnóstica , Terapia por Exercício/instrumentação , Feminino , Humanos , Masculino , Saúde Mental , Autoimagem , Taiwan
14.
J Biol Chem ; 293(1): 324-332, 2018 01 05.
Artigo em Inglês | MEDLINE | ID: mdl-29150441

RESUMO

RcnR, a transcriptional regulator in Escherichia coli, derepresses the expression of the export proteins RcnAB upon binding Ni(II) or Co(II). Lack of structural information has precluded elucidation of the allosteric basis for the decreased DNA affinity in RcnR's metal-bound states. Here, using hydrogen-deuterium exchange coupled with MS (HDX-MS), we probed the RcnR structure in the presence of DNA, the cognate metal ions Ni(II) and Co(II), or the noncognate metal ion Zn(II). We found that cognate metal binding altered flexibility from the N terminus through helix 1 and modulated the RcnR-DNA interaction. Apo-RcnR and RcnR-DNA complexes and the Zn(II)-RcnR complex exhibited similar 2H uptake kinetics, with fast-exchanging segments located in the N terminus, in helix 1 (residues 14-24), and at the C terminus. The largest difference in 2H incorporation between apo- and Ni(II)- and Co(II)-bound RcnR was observed in helix 1, which contains the N terminus and His-3, and has been associated with cognate metal binding. 2H uptake in helix 1 was suppressed in the Ni(II)- and Co(II)-bound RcnR complexes, in particular in the peptide corresponding to residues 14-24, containing Arg-14 and Lys-17. Substitution of these two residues drastically affected DNA-binding affinity, resulting in rcnA expression in the absence of metal. Our results suggest that cognate metal binding to RcnR orders its N terminus, decreases helix 1 flexibility, and induces conformational changes that restrict DNA interactions with the positively charged residues Arg-14 and Lys-17. These metal-induced alterations decrease RcnR-DNA binding affinity, leading to rcnAB expression.


Assuntos
Cobalto/metabolismo , DNA Bacteriano/metabolismo , Proteínas de Escherichia coli/metabolismo , Escherichia coli/metabolismo , Níquel/metabolismo , Proteínas Repressoras/metabolismo , Sequência de Aminoácidos , Sítios de Ligação , Cátions Bivalentes/metabolismo , Proteínas de Ligação a DNA/metabolismo , Escherichia coli/genética , Proteínas de Escherichia coli/química , Espectrometria de Massas , Proteínas Repressoras/química , Relação Estrutura-Atividade , Fatores de Transcrição/metabolismo
15.
Inorg Chem ; 56(11): 6459-6476, 2017 Jun 05.
Artigo em Inglês | MEDLINE | ID: mdl-28517938

RESUMO

Escherichia coli RcnR (resistance to cobalt and nickel regulator, EcRcnR) is a metal-responsive repressor of the genes encoding the Ni(II) and Co(II) exporter proteins RcnAB by binding to PRcnAB. The DNA binding affinity is weakened when the cognate ions Ni(II) and Co(II) bind to EcRcnR in a six-coordinate site that features a (N/O)5S ligand donor-atom set in distinct sites: while both metal ions are bound by the N terminus, Cys35, and His64, Co(II) is additionally bound by His3. On the other hand, the noncognate Zn(II) and Cu(I) ions feature a lower coordination number, have a solvent-accessible binding site, and coordinate protein ligands that do not include the N-terminal amine. A molecular model of apo-EcRcnR suggested potential roles for Glu34 and Glu63 in binding Ni(II) and Co(II) to EcRcnR. The roles of Glu34 and Glu63 in metal binding, metal selectivity, and function were therefore investigated using a structure/function approach. X-ray absorption spectroscopy was used to assess the structural changes in the Ni(II), Co(II), and Zn(II) binding sites of Glu → Ala and Glu → Cys variants at both positions. The effect of these structural alterations on the regulation of PrcnA by EcRcnR in response to metal binding was explored using LacZ reporter assays. These combined studies indicate that while Glu63 is a ligand for both metal ions, Glu34 is a ligand for Co(II) but possibly not for Ni(II). The Glu34 variants affect the structure of the cognate metal sites, but they have no effect on the transcriptional response. In contrast, the Glu63 variants affect both the structure and transcriptional response, although they do not completely abolish the function of EcRcnR. The structure of the Zn(II) site is not significantly perturbed by any of the glutamic acid variations. The spectroscopic and functional data obtained on the mutants were used to calculate models of the metal-site structures of EcRcnR bound to Ni(II), Co(II), and Zn(II). The results are interpreted in terms of a switch mechanism, in which a subset of the metal-binding ligands is responsible for the allosteric response required for DNA release.


Assuntos
Cobalto/metabolismo , Proteínas de Escherichia coli/metabolismo , Ácido Glutâmico/metabolismo , Níquel/metabolismo , Compostos Organometálicos/metabolismo , Proteínas Repressoras/metabolismo , Sítios de Ligação , Cobalto/química , Proteínas de Escherichia coli/genética , Ácido Glutâmico/química , Ligantes , Modelos Moleculares , Níquel/química , Compostos Organometálicos/química , Proteínas Repressoras/genética
16.
J Am Geriatr Soc ; 65(2): 356-363, 2017 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-27879982

RESUMO

OBJECTIVES: To test the effects of a 15-month wheelchair-bound resistance band exercise program on depression and behavioral problems of wheelchair-bound older adults with dementia. DESIGN: Single-blind, cluster-randomized controlled trial with repeated measures. SETTING: Eight nursing homes in southern Taiwan. PARTICIPANTS: Wheelchair-bound nursing home older adults with dementia who participated voluntarily (N = 150) were cluster-randomized to two groups (experimental or control group); 127 completed the study (experimental: four nursing homes, n = 65; control: four nursing homes, n = 62). INTERVENTION: The resistance band exercises were conducted three times per week in 40-minute sessions in the following two sequences: volunteer-led sessions for the first 6 months (Stage I) followed by DVD-guided sessions for the next 9 months (Stage II). MEASUREMENTS: Depression, as measured using the Cornell Scale for Depression in Dementia, and behavioral problems, as measured using the Clifton Assessment Procedures for the Elderly-Behavior Rating Scale, of participants were observed at six time points at 3-month intervals: pretest, two posttests at Stage I, and three posttests at Stage II. RESULTS: By the sixth month of the study, experimental group participants were significantly less depressed and had fewer behavioral problems than control group participants (all P < .05). These small but statistically significant differences persisted throughout the 9 months of the DVD-guided sessions (all P < .05). CONCLUSION: Volunteer-led sessions followed by DVD-guided sessions of resistance band exercise is effective and practical in institutional settings.


Assuntos
Demência/psicologia , Demência/terapia , Depressão/terapia , Comportamento Problema/psicologia , Treinamento Resistido , Idoso , Idoso de 80 Anos ou mais , Pessoas com Deficiência , Feminino , Humanos , Masculino , Casas de Saúde , Método Simples-Cego , Taiwan , Cadeiras de Rodas
17.
Am J Phys Med Rehabil ; 95(11): 789-799, 2016 11.
Artigo em Inglês | MEDLINE | ID: mdl-27149585

RESUMO

OBJECTIVE: The purpose of this study was to test the effects of a 6-month Wheelchair-bound Senior Elastic Band (WSEB) exercise program on the activities of daily living (ADL) and functional fitness of wheelchair-bound older adults with cognitive impairment. DESIGN: Cluster randomized controlled trial was used. A convenience sample of 138 wheelchair-bound older adults with cognitive impairment were recruited from 8 nursing homes in southern Taiwan and were randomly assigned based on the nursing homes they lived to the experimental (4 nursing homes; n = 73) or the control group (4 nursing homes; n = 65). The experimental group performed WSEB exercises 3 times per week and 40 minutes per session for 6 months. The ADL and functional fitness (cardiopulmonary function, body flexibility, range of joint motion, and muscle strength and endurance) were examined at baseline, 3 months, and the end of 6-month study. RESULTS: The ADL and functional fitness indicators of participants in the experimental group showed significant improvements compared to the control group (all P < 0.05). CONCLUSIONS: The WSEB exercises have positive benefits for the ADL and functional fitness of wheelchair-bound older adults with cognitive impairment. It is suggested that WSEB exercises be included as a routine activity in nursing homes. TO CLAIM CME CREDITS: Complete the self-assessment activity and evaluation online at http://www.physiatry.org/JournalCMECME OBJECTIVES:: Upon completion of this article the reader should be able to: (1) Understand the risk factors for functional decline in older adults with dementia; (2) Articulate the benefits of structured activities and exercises in the older adult with dementia; and (3) Incorporate elastic band exercises into the treatment plan of wheelchair bound older adults with dementia. LEVEL: AdvancedACCREDITATION:: The Association of Academic Physiatrists is accredited by the Accreditation Council for Continuing Medical Education to provide continuing medical education for physicians. The Association of Academic Physiatrists designates this activity for a maximum of 1.5 AMA PRA Category 1 Credit(s)™. Physicians should only claim credit commensurate with the extent of their participation in the activity.


Assuntos
Atividades Cotidianas , Transtornos Cognitivos/fisiopatologia , Transtornos Cognitivos/reabilitação , Força Muscular/fisiologia , Treinamento Resistido/métodos , Cadeiras de Rodas , Idoso , Idoso de 80 Anos ou mais , Análise por Conglomerados , Estudos de Coortes , Feminino , Humanos , Masculino , Taiwan , Resultado do Tratamento
18.
J Nurs Res ; 24(2): 137-44, 2016 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-26258390

RESUMO

BACKGROUND: Studies indicate that the incidence of disease, the mortality rate, and medical costs are significantly higher in people aged 65 years and over who do not engage in physical activities than in their peers who do engage in these activities. Therefore, promoting appropriate physical activity among older adults in the community is essential to delaying the health implications of aging. PURPOSE: This pilot test was developed to assess the effectiveness of a newly developed Senior Elastic Band (SEB) exercise program on the health of older adults in community care stations. METHODS: A quasiexperimental design was used. A convenience sample of 20 participants from a community care station was recruited. The SEB intervention included three phases (warm-up, aerobic motion, and static stretching) and was conducted three times per week, 40 minutes per session for 1 month. Twelve health indicators in three categories (functional fitness, self-perceived health status, and sleep quality) were examined before and immediately after 1 month of SEB exercises. RESULTS: Participants showed improved performance at the end of the 1-month study for the following indicators: lung capacity, cardiopulmonary fitness, upper and lower body flexibilities, upper limb muscle power, lower limb muscle endurance, and self-perceived physical health status (all ps < .05). No significant differences were identified for the other indicators. CONCLUSIONS/IMPLICATIONS FOR PRACTICE: The SEB exercise program shows preliminary and promising effects on improving the health of older adults in a community care station. Healthcare professionals who work with older adults living in the community may consider the SEB exercise program as a health promotion modality to recommend and implement with this population. However, we recommend further testing the long-term effects of this program on a larger population.


Assuntos
Exercício Físico , Promoção da Saúde/métodos , Idoso , Idoso de 80 Anos ou mais , Feminino , Nível de Saúde , Humanos , Masculino , Projetos Piloto , Taiwan
19.
Int J Nurs Stud ; 55: 4-14, 2016 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-26655368

RESUMO

BACKGROUND: Physical activity holds promise for mobility-impaired older adults to prevent further disabilities and improve their health. However, staffing constraints have made it challenging to promote physical activity in long-term care facilities. OBJECTIVES: To test the feasibility and effects of 12 months Wheelchair-bound Senior Elastic Band (WSEB) group-exercises that were led by volunteers for the first six months followed by the DVD-guided for another six months on functional fitness, activities of daily living (ADL), and sleep quality of nursing home older adults in wheelchairs. DESIGN: Cluster randomized controlled trial with two groups, pre-test and post-tests. SETTINGS: Ten nursing homes, Taiwan. PARTICIPANTS: 127 participants participated voluntarily; 107 of them completed the study. INCLUSION CRITERIA: (1) aged 65 years and over, (2) using wheelchairs for mobility, (3) living in facility for at least three months, (4) cognitively intact, and (5) heavy or moderate dependency in ADL. Majority of participants were middle-old older adults (75-84 years old, 53.2%), female (51.4%), and had chronic illnesses (98.1%). METHODS: Participants were randomly assigned by facility to either the experimental (five nursing homes, n=56) or control group (five nursing homes, n=51). The WSEB program was conducted three times per week and 40 min per session in two stages: volunteer-led for the first six months (stage I) followed by the DVD-guided modality for another six months (stage II). The primary outcomes (functional fitness: lung capacity, body flexibility, range of joint motion, and muscle strength and endurance) and the secondary outcomes (ADL measured by the Barthel Index; sleep quality measured by the Pittsburgh Sleep Quality Index) of the participants were measured at three time points: pre-test, at the six-month interval, and at the end of 12 months of the study. No blinding was applied. RESULTS: All of the functional fitness indicators of the experimental group participants improved significantly (p<.05), and were all better than the control group at six-month and 12-month of the study (p<.05). No symptoms of discomfort occurred during interventions. CONCLUSIONS: Nursing home older adults in wheelchairs who received WSEB exercise training had better functional fitness, ADL, and sleep quality than those who did not. It was a feasible way of carrying out this exercise program by using the volunteer-led followed by the DVD-guided modalities. The program can be applied in institutional settings routinely.


Assuntos
Exercício Físico , Pacientes Internados , Casas de Saúde , Cadeiras de Rodas , Idoso , Análise por Conglomerados , Estudos de Viabilidade , Humanos , Taiwan
20.
J Adv Nurs ; 71(10): 2338-49, 2015 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-26059214

RESUMO

AIMS: The transtheoretical model was applied to promote behavioural change and test the effects of a group senior elastic band exercise programme on the functional fitness of community older adults in the contemplation and preparation stages of behavioural change. BACKGROUND: Forming regular exercise habits is challenging for older adults. The transtheoretical model emphasizes using different strategies in various stages to facilitate behavioural changes. DESIGN: Quasi-experimental design with pre-test and post-tests on two groups. METHODS: Six senior activity centres were randomly assigned to either the experimental or control group. The data were collected during 2011. A total of 199 participants were recruited and 169 participants completed the study (experimental group n = 84, control group n = 85). The elastic band exercises were performed for 40 minutes, three times per week for 6 months. The functional fitness of the participants was evaluated at baseline and at the third and sixth month of the intervention. Statistical analyses included a two-way mixed design analysis of variance, one-way repeated measures analysis of variance and an analysis of covariance. RESULTS: All of the functional fitness indicators had significant changes at post-tests from pre-test in the experimental group. The experimental group had better performances than the control group in all of the functional fitness indicators after three months and 6 months of the senior elastic band exercises. CONCLUSION: The exercise programme provided older adults with appropriate strategies for maintaining functional fitness, which improved significantly after the participants exercising regularly for 6 months.


Assuntos
Terapia por Exercício/métodos , Idoso , Feminino , Promoção da Saúde/métodos , Humanos , Masculino , Modelos Teóricos , Aptidão Física/fisiologia
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