RESUMEN
Healthcare in inpatient long-term care facilities (nursing homes) should not be limited to medical curative measures, but should also include strengthening social participation, autonomy, self-responsibility and joint responsibility of the residents. Prevention and rehabilitation should therefore be even more integrated into care concepts.This article first introduces various areas of prevention physical activity, nutrition, cognitive competence, psychosocial health, abuse, and freedom-removing measures and then discusses their evidence. Essential for the implementation and the success of such measures is the ability and willingness of people in need of care to engage actively in these therapies; here, appropriate and motivating information plays an important role.Subsequently, geriatric rehabilitation is referred to. In the 2013-2017 empirical study Organization and Rehabilitation for Residents in the Nursing Home to Improve Independence and Participation (ORBIT), 215 people in need of care participated in three-month therapeutic interventions, which were followed by three-months of rehabilitative care. Improvements in mobility and quality of life (Barthel index, QOL-AD) could be demonstrated compared to a control group (nâ¯= 28). The results have to be considered against the background of a worsening health and reduced functional capacities in old age. A stronger integration of prevention and rehabilitation services into long-term institutional care is functional for strengthening participation and independence - an important condition for the residents' certainty that their dignity will be respected, competence and strive for self-responsibility and self-determination.
Asunto(s)
Cuidados a Largo Plazo , Calidad de Vida , Anciano , Anciano de 80 o más Años , Atención a la Salud , Alemania , Humanos , Casas de SaludRESUMEN
In the present work the complexity in the 2D-gel protein pattern of murin lenticular alphaA-Crystallin was analyzed. An in depth study of the different protein isoforms was done combining different proteomic tools. Lens proteins of four different ages, from embryo to 100-week-old mice, were separated by large 2D-PAGE, revealing an increase in the number and intensity of the spots of alphaA-Crystallin during the process of aging. For further analyses the oldest mice were chosen. Comparison and evaluation of two different staining methods proved Imidazole-Zinc to be a good alternative to the generally used Coomassie stain. The characterization of the different alphaA-Crystallin protein species was done using nanoLC-ESI-MS/MS (liquid chromatography electrospray ionisation tandem mass spectrometry). Data interpretation was done by database searching, manual validation and a new MS/MS-interpretation tool for posttranslational modifications--the PTM-Explorer. Using this way, eight different phosphorylation sites were identified and localized; the identification of four of them was not published so far. Furthermore, quantitative N-terminal acetylation of alphaA-Crystallin and variable C-terminal truncation was observed, also not published in this extent yet. The results of the mass spectrometric analysis were validated by immunoblotting experiments using two different alphaA-Crystallin specific antibodies. In addition, a fluorescent phospho-specific stain was used to detect the protein spots including phosphorylation groups. Re-separation 2D-PAGE was done to round off the present study and explain the appearance of some of the protein spots in the gel as artifacts of the 2D-PAGE separation.
Asunto(s)
Procesamiento Proteico-Postraduccional , Proteómica/métodos , Cadena A de alfa-Cristalina/metabolismo , Envejecimiento , Secuencia de Aminoácidos , Animales , Cromatografía Liquida , Electroforesis en Gel Bidimensional/métodos , Imidazoles , Immunoblotting , Cristalino/embriología , Cristalino/crecimiento & desarrollo , Cristalino/metabolismo , Ratones , Ratones Endogámicos C57BL , Datos de Secuencia Molecular , Fosfoproteínas/análisis , Fosforilación , Colorantes de Rosanilina , Espectrometría de Masa por Ionización de Electrospray , Coloración y Etiquetado , Espectrometría de Masas en Tándem , ZincRESUMEN
Dunning R3327 AT-1 rat prostate tumor cells were transfected with a double-fusion suicide gene (CDglyTK) that coded for the cytosine deaminase from E. coli and the thymidine kinase (TK) from HSV-1. The resulting cell line AT-1/CDglyTK was incubated with 10 and 20 microg/ml 5-FC or 0.25 microg/ml GCV, or both 5-FC and GCV 96 hours before harvest. The MTS assay detected cell viabilities of 50+/-5 and 25+/-5% after 5-FC treatment, and 50+/-5% after GCV treatment. The dye exclusion and the colony-forming assay confirmed the data of the MTS assay with GCV (47+/-5 and 32+/-5%), but presented different results for the 5-FC incubation. We detected 100+/-1 and 85+/-5% viable cells after 10 microg/ml 5-FC, and 97+/-1 and 85+/-5% after 20 microg/ml 5-FC treatment, respectively. S-phase arrest in both suicide gene systems was noticeable and a significant increase in cell granularity was observed after incubation with GCV or GCV & 5-FC. This study demonstrates that 5-FC and the metabolized 5-FU act not only as genotoxic reagents, but also as RNA-directed agent, because of the recovery of the cells. On the other hand, a significant S-phase block could be observed after 24 hours incubation with GCV. This short time is enough to incorporate the genotoxic GCV metabolites in the nascent DNA to impair the cell cycle.
Asunto(s)
Citosina Desaminasa/genética , Genes Transgénicos Suicidas , Terapia Genética/métodos , Timidina Quinasa/genética , Animales , Apoptosis , Ciclo Celular , Línea Celular Tumoral , Supervivencia Celular , Citosina Desaminasa/metabolismo , Escherichia coli/genética , Flucitosina/metabolismo , Flucitosina/toxicidad , Ganciclovir/metabolismo , Ganciclovir/toxicidad , Técnicas de Transferencia de Gen , Herpesvirus Humano 1/genética , Masculino , Neoplasias de la Próstata/patología , Ratas , Proteínas Recombinantes de Fusión/genética , Sales de Tetrazolio/metabolismo , Tiazoles/metabolismo , Timidina Quinasa/metabolismo , Pruebas de Toxicidad/métodos , Ensayo de Tumor de Célula MadreRESUMEN
BACKGROUND: Chronic heart failure is a multifactorial, progressive disease of many causes and is associated with complex ventricular remodeling. Deposition of extracellular matrix proteins and sarcomeric disarray of the myocytes occur in end-stage heart failure. Ventricular assist devices (VAD), implanted as bridge to transplantation, may reverse ventricular remodeling. Although successfully weaning patients from VAD support has been reported, it is not clear to what degree reversal of remodeling occurs in unloaded failing hearts. Because collagen deposition and ultrastructural disarray are hallmarks of myocardial remodeling, we analyzed the myocardial ultrastructure and collagen content of VAD-supported hearts before and after mechanical unloading. METHODS: We used amino acid analysis to measure collagen content (4-hydroxyproline content) in 24 transplant candidates receiving VAD support. We used transmission electron microscopy to examine the ultrastructure in 6 patients receiving VAD support. RESULTS: The 4-hydroxyproline content increased significantly at VAD implantation and was not altered by mechanical unloading. The ultrastructure showed signs of persisting cardiomyopathy. CONCLUSION: Mechanical unloading does not alter the total collagen content of the supported, failing heart. Thus, structural reversal of the remodeling process associated with heart failure is not a general phenomenon in mechanically unloaded hearts.
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Insuficiencia Cardíaca/terapia , Corazón Auxiliar , Hidroxiprolina/metabolismo , Miocardio/metabolismo , Miocardio/ultraestructura , Remodelación Ventricular/fisiología , Adolescente , Adulto , Anciano , Niño , Colágeno/metabolismo , Insuficiencia Cardíaca/metabolismo , Insuficiencia Cardíaca/patología , Trasplante de Corazón , Ventrículos Cardíacos/metabolismo , Ventrículos Cardíacos/ultraestructura , Humanos , Microscopía Electrónica , Persona de Mediana EdadRESUMEN
Relapse with drug-resistant disease is the main cause of death in MYCN-amplified neuroblastoma patients. MYCN-amplified neuroblastoma cells in vitro are characterized by a failure to arrest at the G(1)-S checkpoint after irradiation- or drug-induced DNA damage. We show that several MYCN-amplified cell lines harbor additional chromosomal aberrations targeting p53 and/or pRB pathway components, including CDK4/CCND1/MDM2 amplifications, p16INK4A/p14ARF deletions or TP53 mutations. Cells with these additional aberrations undergo significantly lower levels of cell death after doxorubicin treatment compared with MYCN-amplified cells, with no additional mutations in these pathways. In MYCN-amplified cells CDK4 expression is elevated, increasing the competition between CDK4 and CDK2 for binding p21. This results in insufficient p21 to inhibit CDK2, leading to high CDK4 and CDK2 kinase activity upon doxorubicin treatment. CDK4 inhibition by siRNAs, selective small compounds or p19(INK4D) overexpression partly restored G(1)-S arrest, delayed S-phase progression and reduced cell viability upon doxorubicin treatment. Our results suggest a specific function of p19(INK4D), but not p16(INK4A), in sensitizing MYCN-amplified cells with a functional p53 pathway to doxorubicin-induced cell death. In summary, the CDK4/cyclin D-pRB axis is altered in MYCN-amplified cells to evade a G(1)-S arrest after doxorubicin-induced DNA damage. Additional chromosomal aberrations affecting the p53-p21 and CDK4-pRB axes compound the effects of MYCN on the G(1) checkpoint and reduce sensitivity to cell death after doxorubicin treatment. CDK4 inhibition partly restores G(1)-S arrest and sensitizes cells to doxorubicin-mediated cell death in MYCN-amplified cells with an intact p53 pathway.
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Quinasa 4 Dependiente de la Ciclina/antagonistas & inhibidores , Daño del ADN/efectos de los fármacos , Doxorrubicina/toxicidad , Proteínas Nucleares/metabolismo , Proteínas Oncogénicas/metabolismo , Apoptosis , Línea Celular Tumoral , Aberraciones Cromosómicas , Ciclina D1/metabolismo , Quinasa 2 Dependiente de la Ciclina/antagonistas & inhibidores , Quinasa 2 Dependiente de la Ciclina/genética , Quinasa 2 Dependiente de la Ciclina/metabolismo , Quinasa 4 Dependiente de la Ciclina/genética , Quinasa 4 Dependiente de la Ciclina/metabolismo , Inhibidor p19 de las Quinasas Dependientes de la Ciclina/metabolismo , Inhibidor p21 de las Quinasas Dependientes de la Ciclina/metabolismo , Puntos de Control de la Fase G1 del Ciclo Celular/efectos de los fármacos , Humanos , Proteína Proto-Oncogénica N-Myc , Neuroblastoma/metabolismo , Neuroblastoma/patología , Proteínas Nucleares/genética , Proteínas Oncogénicas/genética , Interferencia de ARN , ARN Interferente Pequeño/metabolismo , Proteína de Retinoblastoma/metabolismo , Puntos de Control de la Fase S del Ciclo Celular/efectos de los fármacos , Proteína p53 Supresora de Tumor/genética , Proteína p53 Supresora de Tumor/metabolismoRESUMEN
Proteome studies with small sample amounts are difficult to perform, especially when membrane proteins are the focus of interest. In our study a new method for the analysis of scarce membrane protein samples combining large gel 2-D-CTAB/SDS-PAGE with fluorescence dye saturation labelling (satDIGE) was developed, allowing a highly sensitive differential analysis of different cell states. After Triton X-114 phase partitioning, enriched membrane protein samples of T cells were labelled at cysteine residues using fluorescence dyes and separated by large gel 2D-CTAB/SDS-PAGE. For a differential analysis 3 mug protein was found to be sufficient to detect proteins in a widespread well-separated diagonal spot pattern.