A novel procedure for protein extraction from formalin-fixed paraffin-embedded tissues.

Most of the archived pathological specimens in hospitals are kept as formalin-fixed paraffin-embedded tissues (FFPE) for long-term preservation. Up to now, these samples are only used for immunohistochemistry in a clinical routine as it is difficult to recover intact protein from these FFPE tissues. Here, we report a novel, short time-consuming and cost-effective method to extract full-length, non-degraded proteins from FFPE tissues. This procedure is combined with an effective and non-toxic deparaffinisation process and an extraction method based on antigen-retrieval, high concentration of SDS and high temperature. We have obtained enough intact protein to be detected by Western blotting analysis. This technique will allow utilising these stored FFPE tissues in several applications for protein analysis helping to advance the translational studies in cancer and other diseases.

Stem cell therapy in chronic ischemic heart dysfunction with and without viability.

A growing number of clinical trials are evaluating the effects of stem cell therapy in patients with chronic ischemic heart dysfunction. As most of the clinical trials included a limited and different number of patients, various stem cell sources and several delivery approaches, results vary substantially between these studies. We analyse whether the assessment of myocardial viability may be important when evaluating effects of stem cell transplantation on parameters of left ventricular remodeling. Viability assessment could help to find the best type of stem cell and the best method of cell delivery to be used in chronic ischemic heart dysfunction.

Human CD8+ T cell blasts are more sensitive than CD4+ T cell blasts to regulation by APO2L/TRAIL.

The mechanisms responsible for the down-modulation of the activation of separated CD4(+) or CD8(+) human T cell blasts were studied using cells obtained from healthy donors. In the presence of IL-2, human CD8(+) T cell blasts were more sensitive than CD4(+) T cell blasts to regulation by APO2 ligand/TNF-related apoptosis-inducing ligand (APO2L/TRAIL), while both T cell subsets were equally sensitive to Fas/CD95 regulation. This regulation was defined as inhibition of IL-2-dependent T cell growth in the absence of cell death induction, characterized by cell cycle arrest in G(2)/M. The physiological validity of these observations was corroborated by the demonstration of intracellular FasL and APO2L/TRAIL expression in CD4(+) and CD8(+) T cell blasts, which were secreted in their bioactive form into the supernatant upon PHA, CD3 or CD59 reactivation. Additionally, the inhibition of IL-2-dependent CD4(+) or CD8(+) T cell blast growth upon CD3 or CD59 ligation was dependent, at least partially, on FasL and/or APO2L/TRAIL. These data precisely define the role of APO2L/TRAIL in the regulation of human T cell activation.

Efectividad de la intervención educativa en cuidadores de pacientes dependientes en diálisis y valoración de la carga / The effectiveness of educational intervention in carers of patients in dialysis and evaluation of the load

Los cambios demográficos y el envejecimiento paulatino dela población con insuficiencia renal crónica en tratamiento renal sustitutivo, así como las múltiples patologías asociadas determinan una población muy dependiente de un cuidador. Por todo esto, evaluamos la carga y grado de conocimientos del cuidador principal de los pacientes “dependientes” en tratamiento renal sustitutivo del Complexo Hospitalario de Ourense. Con los resultados obtenidos realizamos una intervención educativa, valorando posteriormente su efectividad. Se determinó que pacientes son dependientes en su tratamiento y autocuidados y se realizó un estudio de sus cuidadores. Se aplicó la escala sobre carga del cuidador de Zarity Zarit y la escala sobre conocimientos del cuidador principal, antes y después de la intervención educativa. En nuestra población el 24,5 % de los pacientes en HD y el 33,3 % en DP son dependientes de un cuidador. Los cuidadores no percibían sobrecarga, siendo el grado similar en ambos tipos de diálisis (hospitalaria vs domiciliaria) y además esta percepción disminuía con el tiempo como cuidador. El grado de conocimientos del cuidador aumentó significativamente tras la intervención educativa. Es importante dar una información adecuada al inicio del tratamiento y continuarla a lo largo del mismo, ya que por sí solos los cuidadores no aumentan el nivel de conocimientos

Good education improves quality of life; the demographic changes and the gradual ageing of the population with chronicrenal insufficiency in substitutive renal treatment, and the many associated pathologies determine a populationthat is highly dependent on carers. We therefore evaluate the load and knowledge of the main carers of “dependent “patients in substitutive renal treatment in the Ourense Hospital Complex. With the results, we perform an educational intervention and follow up by evaluating its effectiveness. It is determined which patients are dependent in their treatment and those who look after themselves, and a study is made of the carers. The Zarit and Zarit scale is applied to the load of the carer and the knowledge of the main care before and after the education. In our population, 24.5 % of the patients in HD and 33.3% in DP are dependent on a carer. The carers perceived no overloading, and the load was similar in both types of dialysis (hospital v home), this perception also falling with their time as carers. The carers’ knowledge increases significantly following the intervention; suitable information must be given at the beginning of the treatment, and must be continued throughout, as the carers do not increase their knowledge without help