Isolation and morphological characterization of equine mesenchymal stem cells from harvested adipose tissue and bone marrow and stably transfected with green fluorescent protein.

OBJECTIVE: To characterize the ultrastructure of mesenchymal stem cells (MSCs) that were harvested from the adipose tissue (AT-MSCs) and bone marrow (BM-MSCs) of horses and transfected with green fluorescent protein. SAMPLE: MSCs from adipose tissue and bone marrow of 6 adult female Hispano-Bretón horses. PROCEDURES: Harvested equine MSCs were cultivated and transfected with green fluorescent protein, and the immunophenotypes of the MSCs were characterized by use of anti-CD90 and anti-CD105 monoclonal antibodies. When stable transfection of MSCs was achieved, the morphological and ultrastructural characteristics of transfected and nontransfected AT-MSCs and BM-MSCs were compared with electron microscopy. RESULTS: The protocols for transfection and subsequent isolation of transfected cells with use of G418 were suitable for obtaining transfected MSCs. Transfection efficiency was 5% in AT-MSCs and 4% in BM-MSCs. Characterization of transfected and nontransfected MSCs revealed that they share immunocytochemical and morphological profiles. Expression of CD90 was significantly higher for transfected versus nontransfected AT-MSCs (97% vs 92%). Expression of CD105 was significantly lower for transfected versus nontransfected BM-MSCs (85% vs 94%). Transfected BM-MSCs had differences in organelles, compared with the other cell types, specifically including most commonly the rough endoplasmic reticulum with dilated cisternae and mitochondria. CONCLUSION AND CLINICAL RELEVANCE: These findings contribute to the knowledge base of the characteristics of equine AT-MSCs and BM-MSCs and of transfected versus nontransfected equine MSCs. The data provided a valuable starting point for researchers wishing to further study the morphological characteristics of equine MSCs.

Effect of mesenchymal stem cells combined with chondroitin sulfate in an in vitro model of osteoarthritis.

Osteoarthritis (OA) is a degenerative joint disease affecting the whole joint structure. The specific molecules responsible for the inflammatory processes involved in the development of OA have been the focus of many studies. Adipose tissue-derived mesenchymal stem cells (ASCs) constitute a promising cell-based therapy which could be used as an alternative to or in combination with drug therapies. Chondroitin sulfate (CS) plays a protective role in the joint by decreasing concentrations of pro-inflammatory cytokines and therefore has an important part in moderating chondrocyte metabolism. The aim of this study is to use an in vitro model of OA to evaluate the combined effectiveness of CS and ASCs as a treatment. We give a detailed discussion of the roles of cytokines and other key molecules involved in OA. In addition, we report the effects of treating inflamed chondrocytes with ASCs and CS on the expression of specific cartilage genes. Findings show that both treatments reduced expression of all genes associated with the pro-inflammatory cytokines we analyzed. However, we saw no increase in the expression of the specific genes encoding for cartilage matrix proteins, such as collagen type II and aggrecan. This study shows the effectiveness of combining ASCs and CS in the treatment of OA.

Access to Information and Degree of Community Awareness of Preventive Health Measures in the Face of COVID-19 in Spain.

The COVID-19 pandemic is posing a major health crisis. Spanish legislation establishes the mandatory use of masks and the implementation of hygienic measures such as hand washing and physical distancing. The aim of this study is to describe access to information and the level of community knowledge/adoption about the preventive measures proposed by the Spanish health authorities in response to the COVID-19 pandemic and to analyze the influence of socio-demographic factors in compliance among people over 18 years of age resident in Spain. An observational, descriptive and transversal study was conducted. Data was collected on sociodemographic variables, access to information and the degree of knowledge/adoption about the preventive measures: use of masks, hand hygiene and physical distancing. A total of 1811 people participated. The average age was 45.1 ± 15.1 years, predominantly female (69.3%), from an urban geographical area (74%), with a higher education level of 53.2%. Most of the respondents (57.5%) are or live with people at risk. The main access to information on preventive measures was from secondary sources (49.2%), with television being the main medium; 72.3% think that there are some difficulties in accessing information, while 8.7% of the participants do not consider the use of masks to be useful. As regards the choice of type of mask, the majority of people (44.8%) opt for the surgical variety; 88.5% of respondents believe that the physical distancing established is at least 1.5 m. This study confirmed that socio-demographic factors influence compliance with or the degree of knowledge/adoption of the preventive measures proposed to combat the COVID-19 pandemic and has made it possible to ascertain the sectors of the population with the greatest deficiencies in this respect. It shows the importance of implementing health information and education systems in the community, and it is advisable to promote specific programs aimed at men, people living in rural areas and people with a low level of education.

Isolation and characterization of exosomes from adipose tissue-derived mesenchymal stem cells.

Mesenchymal stem cells (MSCs) are the subject of intense research as they are a potential therapeutic tool for several clinical applications. The new MSCs action models are focused on the use of MSC-derived secretome which contains several growth factors, cytokines, microRNAs, and extracellular vesicles such as exosomes. Exosomes have recently emerged as a component with great potential involved as mediators in cellular communication. The isolation and identification of exosomes has made it possible for them to be used in cell-free therapies. The purposes of this study are: (i) to detect exosomes released into adipose-derived MSC conditioned cell culture medium, (ii) to identify exosome morphology, and (iii) to carry out a complete characterization of said exosomes. Moreover, it is aimed at determining which method for exosome isolation would be best to use. Precipitation has been identified as a highly useful method of exosome isolation since it provides higher efficiency and purity values than other methods. A broad characterization of the exosomes present in the MSC-conditioned medium was also carried out. This work fills a gap in the existing literature on bioactive molecules which have attracted a great deal of interest due to their potential use in cellular therapies.

Calidad de vida en pacientes con insuficiencia cardíaca crónica. Influencia del tratamiento / Quality of life in patients with chronic heart failure. The role of treatment

Introducción. La insuficiencia cardiaca crónica (ICC) es una de las principales enfermedades crónicas que afectan a la calidad de vida (CV). En nuestro medio, solo existe un estudio realizado en pacientes ambulatorios de Atención Primaria, a priori, clínicamente estables y con un grado funcional bajo. Se plantea la necesidad de medir la CV en pacientes cuyo grado funcional se prevea grave. Objetivo. Describir la calidad de vida en pacientes con insuficiencia cardiaca crónica en una unidad de insuficiencia cardiaca y analizar la asociación con factores terapéuticos. Método. Mediante un estudio transversal se midió la CV con el cuestionario 36-Item Short-Form Health Survey en 87 enfermos con ICC reclutados desde la UIC del Complejo Asistencial Universitario de León. Resultados. Los enfermos presentaron edad avanzada (71.3 años ±13.3); predominó el sexo masculino (70.6 %); el 94.5 % recibía tratamiento con diuréticos, el 74.1 % con inhibidores de la enzima de conversión de la angiotensina o antagonistas de los receptores de la angiotensina II, el 85.9 % con betabloqueantes y un 21 % portaban un desfibrilador automático implantable; la puntuación total de CV en el componente sumario físico fue de 33.2 ±10,6 y en el componente sumario mental de 48.9 ±13,1. El valor más alto se halló en el rol emocional (45.5) y el más bajo en la función física (33.1). Conclusiones. La percepción de la CV en pacientes con ICC está considerablemente alterada en todas las dimensiones, dentro de las cuales sufren un mayor deterioro las físicas que las emocionales. De todas las drogas estudiadas, la terapia con betabloqueantes es la que supuso un mayor impacto en las dimensiones físicas de la CV de los pacientes, al asociarse de forma independiente su uso con la mejora de esta

Introduction. Chronic heart failure (CHF) is one of the major chronic diseases which affect quality of life (QoL). In our field, only one study has been addressed to study the QoL in outpatient primary care of clinically stable CHF patients with a low degree of functional capacity. There is an imperative need to measure QoL in severe CHF patients. Objective. To describe the QoL of patients suffering from chronic heart failure in hospital’s Heart Failure Unit (HFU) and correlate it to therapeutic factors. Method. A QoL cross-sectional study was performed to 87 patients with CHF, using the 36-Item Short-Form Health Survey. Patients were recruited from the HFU of University Health Care Complex in Leon, Spain. Results. Patients were mostly elderly (13.3 ± 71.3 years), predominantly male (70.6 %). 94.5 % were treated with diuretics, 74.1 % with angiotensin-convertingenzyme inhibitors and/or angiotensin II receptor blockers, 85.9 % were treated with beta-blockers and 21 % had an implantable cardioverter defibrillator. The total QoL score in the physical component summary was 33.2 ± 10.6 and the mental component summary showed a score of 48.9 ± 13.1. The highest value was found in the emotional role (45.5) and the lowest in physical function (33.1). Conclusions. The perceived QoL in patients with CHF is significantly altered in all its dimensions. Patients perceive a stronger physical than emotional deterioration. Among all drugs studied, the betablocker therapy is one that represented greater benefits, impacting positively on patient's QoL physical dimensions

Calidad de vida en pacientes con insuficiencia cardiaca crónica / Quality of life in patients with chronic cardiac failure

OBJETIVO: evaluar la calidad de vida (CV) en pacientes con insuficiencia cardiaca crónica (ICC) pertenecientes a una unidad de insuficiencia cardiaca (UIC) y valorar la asociación con factores clínicos. MÉTODO: estudio descriptivo transversal en 87 enfermos con ICC pertenecientes la UIC del "Complejo Asistencial Universitario de León". Para medir la CV se utilizó el cuestionario 36-Item Short-Form Health Survey (SF-36). La comparación de variables se realizó con el testt-Student. Para evaluar la asociación independiente entre las diferentes dimensiones y las variables, se construyeron modelos de regresión lineal múltiple. RESULTADOS: la edad media de los sujetos estudiados fue de 71,3años, predominó el sexo masculino (70,6%), la etiología no-isquémica(57,6%), el grado funcional de la New York Heart Association (NYHA)III-IV (52,9%) y la IC sistólica (82,4%). La puntuación media de CV en el componente sumario físico (CSF) fue de 33,2 y en el componente sumariomental (CSM) de 48,9. En los análisis multivariantes: para el CSF, un grado funcional III-IV de la NYHA y una etiología no-isquémica, empeoraban la CV en su componente físico; el componente mental de la CV empeoraba en la IC diastólica y con etiología no-isquémica. CONCLUSIONES: la percepción de la CV en pacientes con ICC está considerablemente alterada en todas las dimensiones. Un peor grado funcional y la etiología no isquémica se asociaron independientemente con un mayor deterioro de la CV en las dimensiones físicas, y la IC diastólica y la etiología no isquémica con un mayor deterioro en las emocionales. La etiología es la variable con mayor impacto en todos los dominios

OBJECTIVE: critical patients tend to present hyperglycemia, a fact associated with an increase in morbimortality and infectious complications. According to the Spanish Society of Intensive and Critical Medicine and Coronary Units (SEMICYUC), a safe and good-quality care requires maintaining glycemic levels between80 and 150 mg/dl with insulin therapy. The objective is to obtain an estimation of the adequate maintenance of glycemic levels of patients, as well as to detect any potential for quality improvement, if necessary. METHOD: a descriptive observational study conducted in the Intensive Care Unit of the Hospital Virgen del Rocío during January, February and March, 2014. The sample size was estimated in 108 patients, recruited in 13 days randomly chosen, with all week days represented. The indicator was measured according to the following formula: (Number of patients with glycemia > 150 mg/dl and on insulin treatment / Number of patients with indication for glycemic control and glycemias> 150mg/d) X 100.RESULTS: the mean age of subjects was 64 years, with 73%male and 27% women. A 41% of the sample suffered Type 2diabetes, 1% suffered Type I diabetes, and 58% had no diabetes. A range of glycemias between 151 mg/dl and 323 mg/dl was obtained. The minimum level of the indicator required by the SEMICYUC (80%) was not reached in any of the three months measured, thus being in a sub-standard situation. CONCLUSIONS: there is no adequate maintenance of the glycemi levels in patients managed in our unit; therefore, a review of the current protocol for insulin therapy could improve the quality and safety in this aspect of care