Desarrollo de un instrumento para evaluar la complejidad física de pacientes con secuela de quemaduras: Consenso e-Delphi / Development of an instrument to evaluate the physical complexity of patients with burn sequelae: e-Delphi Consensus

INTRODUCCION: En la literatura existen escalas de valoración de cicatrices, basadas en la observación de características físicas de una porción de ellas, pero ninguna se refiere al nivel de compromiso físico que la secuela produce en el paciente desde el punto de vista del tratante o evaluador. OBJETIVO: Identificar en forma consensuada, elementos determinantes de la complejidad física del paciente con cicatrices e injertos producto de una o más quemaduras. MATERIAL Y MÉTODOS: Quince profesionales médicos, kinesiólogos y terapeutas ocupacionales, que cumplieron diferentes requisitos de selección, participaron en un consenso e-Delphi, para operacionalizar el constructo "complejidad física de un paciente con secuela de quemaduras", constituido por tres rondas de encuestas anónimas, con retroalimentación, cuyos resultados se evaluaron mediante análisis de frecuencia, considerando como aceptable una concordancia de 80% y más. RESULTADOS: Como componentes de la complejidad física del paciente con secuela de quemaduras, se identificaron las dimensiones: localización, edad, etapa de evolución de los injertos, etapa de evolución de las cicatrices, tipo de cicatrices, tipo de injertos, grado de retracción en las secuelas y tipo de deformidad. Cada dimensión, con sus respectivos ítems. CONCLUSION: El consenso fue alcanzado en una variedad de dimensiones y sus ítems específicos para identificar los principales componentes involucrados en la complejidad física del paciente con secuela de quemaduras, lo que permite incluir esta información en una escala, cuya validación biométrica podría favorecer el pronóstico de la evolución de la secuela de quemaduras, orientar la rehabilitación, mejorar la gestión administrativa y calidad de vida del paciente.

INTRODUCTION: In literature there are scales for scar assessment, based on the observation of physical characteristics of each patient´s scar but none refer to the level of physical compromise that the sequelae produces in the patient from the point of view of the treating physician or evaluator. OBJECTIVE: To identify, by consensus, elements that determining the physical complexity of patients with scars and skin grafts resulting from one or more burns.MATERIALS AND METHODS: A total of 15 health care professionals physicians, physiotherapist and occupational therapists, who met different selection requirements, participated in an e-Delphi consensus, to operationalize the construct of "physical complexity of a patient with burn sequelae". The e-Delphi panel consisted of three rounds of anonymous surveys, with feedback, and the results were evaluated by frequency analysis. An 80% or more of concordance was considered as acceptable.RESULTS: The following dimensions were identified as components of physical complexity in patients with burn sequelae; location, age, stage of skin graft progress, type of scar, type of skin graft, retraction degree of sequelae and type of deformity. Each dimension with its respective items.CONCLUSION: The consensus was reached on a variety of dimensions and their specific items to identify the main components involved in the physical complexity in patients with burn sequelae. The biometric validation of this information include on a scale could report regarding the prognosis the burn sequelae, guide rehabilitation therapy, improve the management administration and the quality of life of patient with burn sequelae.

Biochemical characterization of the bifunctional enzyme dihydrofolate reductase-thymidylate synthase from Leishmania ( Viannia ) and its evaluation as a drug target / Caracterización bioquímica de la enzima bifuncional dihidrofolato reductasa-timidilato sintasa de Leishmania ( Viannia ) y su evaluación como blanco molecular

Introduction. Dihydrofolate reductase (DHFR) has been used successfully as a drug target in the area of anti-bacterial, anti-cancer and anti-malarial therapy. Although this bifunctional enzyme is also a potential drug target for treatment of leishmaniasis, there have been no reports on its efficacy against Leishmania ( Viannia ) species . Materials and methods. The gene encoding the bifunctional DHFR and thymidylate synthase (TS) of Le. (V.) braziliensis was isolated and expressed in E. coli. The enzyme was purified and characterized. The inhibitory effects of antifolates and four aporphine alkaloids on its activity were evaluated. Results. The full-length gene consists of a 1560-bp open reading frame encoding a 58 kDa translated peptide containing DHFR and TS domains linked together in a single polypeptide chain. The recombinant DHFR-TS enzyme revealed K m and V max values of 55.35 ± 4.02 µ M (mean ± SE) and 0.02 ± 5.34 x 10 -4 µ M/min respectively for dihydrofolic acid (H 2 F). The Le. braziliensis rDHFR-TS have Ki values for antimicrobial antifolates in the µM range. Methotrexate (MTX) was a more-potent inhibitor of enzymatic activity ( Ki = 22.0 µM) than trimethoprim ( Ki = 33 µM) and pyrimethamine ( Ki = 68 µM). These Ki values are significantly lower than those obtained for the aporphine alkaloids. Conclusion. The results of the study show the inhibitory effect of antifolate drugs on enzymatic activity, indicating that Le. braziliensis rDHFR-TS could be a model to studying antifolate compounds as potential antiprotozoal drugs.

Introducción. La dihidrofolato reductasa (DHFR) se ha utilizado como blanco molecular en tratamientos antibacterianos, anticancerígenos y antipalúdicos. También, actúa como blanco molecular en Leishmania ; sin embargo, no existen reportes de la enzima bifuncional en especies de Leishmania ( Viannia ). Materiales y métodos. Se ha aislado y expresado en Escherichia coli el gen que codifica para la enzima bifuncional DHFR y la timidilato-sintasa (TS) de Leishmania braziliensis . La enzima recombinante se purificó y caracterizó, y se evaluó el efecto inhibitorio de algunos antifolatos, así como de cuatro alcaloides aporfínicos. Resultados. El gen se compone de aproximadamente 1.560 pb y codifica un péptido de 58 kDa que contiene los dominios DHFR y TS ligados en una sola cadena polipeptídica. La enzima recombinante DHFR-TS, utilizando el dihidrofolato (H2F) como sustrato, presentó valores de K m y V max de 55,35 ± 4,02 (media ± el error estándar de la media) y de 0,02 ± 5,34 x 10 -4 , respectivamente. La enzima rDHFR-TS de L. braziliensis presentó valores de Ki para los antifolatos en el rango de micras. El metotrexato fue el inhibidor más potente de la actividad enzimática ( Ki =22,0 mM) en comparación del trimetoprim ( Ki =33 mM) y la pirimetamina ( Ki =68 mM). Estos valores de Ki son significativamente más bajos en comparación con los obtenidos para los alcaloides aporfínicos. Conclusión. Los resultados muestran el efecto inhibitorio de los antifolatos sobre la actividad enzimática, lo cual indica que la rDHFR-TS de L. braziliensis podría ser un modelo para estudiar moléculas antiprotozoarias potenciales.

Clinical and experimental approaches to knee cartilage lesion repair and mesenchymal stem cell chondrocyte differentiation.

Cartilage has poor regeneration capacity due to the scarcity of endogenous stem cells, its low metabolic activity and the avascular environment. Repair strategies vary widely, including microfracture, autologous or allogenic tissue implantation, and in vitro engineered tissues of autologous origin. However, unlike the advances that have been made over more than two decades with more complex organs, including vascular, cardiac or bone tissues, similar advances in tissue engineering for cartilage repair are lacking. Although the inherent characteristics of cartilage tissue, such as the lack of vascularity and low cellular diversity, suggest that it would be one of the more simple tissues to be engineered, its functional weight-bearing role and implant viability and adaptation make this type of repair more complex. Over the last decade several therapeutic approaches and innovative techniques show promise for lasting and functional regeneration of hyaline cartilage. Here we will analyze the main strategies for cartilage regeneration and discuss our experience.

Biochemical characterization of the bifunctional enzyme dihydrofolate reductase-thymidylate synthase from Leishmania (Viannia) and its evaluation as a drug target.

INTRODUCTION: Dihydrofolate reductase (DHFR) has been used successfully as a drug target in the area of anti-bacterial, anti-cancer and anti-malarial therapy. Although this bifunctional enzyme is also a potential drug target for treatment of leishmaniasis, there have been no reports on its efficacy against Leishmania (Viannia) species. MATERIALS AND METHODS: The gene encoding the bifunctional DHFR and thymidylate synthase (TS) of Le. (V.) braziliensis was isolated and expressed in E. coli. The enzyme was purified and characterized. The inhibitory effects of antifolates and four aporphine alkaloids on its activity were evaluated. RESULTS: The full-length gene consists of a 1560-bp open reading frame encoding a 58 kDa translated peptide containing DHFR and TS domains linked together in a single polypeptide chain. The recombinant DHFR-TS enzyme revealed Km and Vmax values of 55.35 ± 4.02 µ M (mean ± SE) and 0.02 ± 5.34 x 10 -4 µ M/min respectively for dihydrofolic acid (H2F). The Le. braziliensis rDHFR-TS have Ki values for antimicrobial antifolates in the µM range. Methotrexate (MTX) was a more-potent inhibitor of enzymatic activity (Ki = 22.0 µM) than trimethoprim (Ki = 33 µM) and pyrimethamine (Ki = 68 µM). These Ki values are significantly lower than those obtained for the aporphine alkaloids. CONCLUSION: The results of the study show the inhibitory effect of antifolate drugs on enzymatic activity, indicating that Le. braziliensis rDHFR-TS could be a model to studying antifolate compounds as potential antiprotozoal drugs.

Clinical and experimental approaches to knee cartilage lesion repair and mesenchymal stem cell chondrocyte differentiation

Cartilage has poor regeneration capacity due to the scarcity of endogenous stem cells, its low metabolic activity and the avascular environment. Repair strategies vary widely, including microfracture, autologous or allogenic tissue implantation, and in vitro engineered tissues of autologous origin. However, unlike the advances that have been made over more than two decades with more complex organs, including vascular, cardiac or bone tissues, similar advances in tissue engineering for cartilage repair are lacking. Although the inherent characteristics of cartilage tissue, such as the lack of vascularity and low cellular diversity, suggest that it would be one of the more simple tissues to be engineered, its functional weight-bearing role and implant viability and adaptation make this type of repair more complex. Over the last decade several therapeutic approaches and innovative techniques show promise for lasting and functional regeneration of hyaline cartilage. Here we will analyze the main strategies for cartilage regeneration and discuss our experience.

Glucose transporter 1 and monocarboxylate transporters 1, 2, and 4 localization within the glial cells of shark blood-brain-barriers.

Although previous studies showed that glucose is used to support the metabolic activity of the cartilaginous fish brain, the distribution and expression levels of glucose transporter (GLUT) isoforms remained undetermined. Optic/ultrastructural immunohistochemistry approaches were used to determine the expression of GLUT1 in the glial blood-brain barrier (gBBB). GLUT1 was observed solely in glial cells; it was primarily located in end-feet processes of the gBBB. Western blot analysis showed a protein with a molecular mass of 50 kDa, and partial sequencing confirmed GLUT1 identity. Similar approaches were used to demonstrate increased GLUT1 polarization to both apical and basolateral membranes in choroid plexus epithelial cells. To explore monocarboxylate transporter (MCT) involvement in shark brain metabolism, the expression of MCTs was analyzed. MCT1, 2 and 4 were expressed in endothelial cells; however, only MCT1 and MCT4 were present in glial cells. In neurons, MCT2 was localized at the cell membrane whereas MCT1 was detected within mitochondria. Previous studies demonstrated that hypoxia modified GLUT and MCT expression in mammalian brain cells, which was mediated by the transcription factor, hypoxia inducible factor-1. Similarly, we observed that hypoxia modified MCT1 cellular distribution and MCT4 expression in shark telencephalic area and brain stem, confirming the role of these transporters in hypoxia adaptation. Finally, using three-dimensional ultrastructural microscopy, the interaction between glial end-feet and leaky blood vessels of shark brain was assessed in the present study. These data suggested that the brains of shark may take up glucose from blood using a different mechanism than that used by mammalian brains, which may induce astrocyte-neuron lactate shuttling and metabolic coupling as observed in mammalian brain. Our data suggested that the structural conditions and expression patterns of GLUT1, MCT1, MCT2 and MCT4 in shark brain may establish the molecular foundation of metabolic coupling between glia and neurons.

Castration induces changes in the cation-dependent mannose-6-phosphate receptor in rat epididymis: possible implications in secretion of lysosomal enzymes.

It is believed that the mammalian epididymis participates in the maturation of the sperm due to its secretory activity. High concentrations of several secreted acid hydrolases are found in the epididymal lumen. Moreover, some of these enzymes are secreted by the epididymal epithelium in an androgen-dependent fashion. In this study, we attempted to discern whether mannose-6-phosphate receptors (MPRs) regulate transport and secretion of lysosomal enzymes in the rat epididymis, and if these events are altered when the animals are subjected to hormonal manipulation. We observed that expression of cation-dependent MPR (CD-MPR) and cation-independent MPR (CI-MPR) increased significantly in caudal epididymis of castrated rats by immunoblot. This increase was corroborated by quantitation of MPRs, by binding assays. This change could be due to androgen deprivation, as a similar effect was observed after treatment with the anti-androgenic drug flutamide. Furthermore, we observed that the CD-MPR was redistributed to the apical area of the epithelium on castrated rats by immunohistochemistry, which is compatible with the redistribution of the receptors toward lighter fractions in a Percoll gradient. Consistent with a possible involvement of the CD-MPR in the secretion, we observed an increase in pro-cathepsin D levels in epididymal fluid after castration. We conclude that the CD-MPR might be regulated by hormones and that this receptor might be involved in the secretion of specific enzymes into the rat epididymis.

La legibilidad de los materiales educativos sobre la diabetes: Implicaciones para la educación de pacientes con materiales escritos / Readability of Diabetes Education Materials: Implications for reaching patients with written materials

Objetivo: Determinar el nivel educativo necesario para comprender los materiales educativos suministrados a pacientes diabéticos por organizaciones gubernamentales y NGOs. Materiales y métodos: Este es un estudio descriptivo que explora la legibilidad de 81 materiales de educación disponibles a pacientes con diabetes y distribuidos por proveedores de salud. Se utilizó 2 medidas para determinar los niveles de legibilidad de los materiales informativos para diabéticos, el SMOG Readability Formula y el Fray Graph. La muestra excluyó materiales educativos que no estuvieran en inglés y aquellos con objetivos comerciales. Para el análisis se utilizó medidas descriptivas y prueba t para muestras y se interpretó el valor de p. Resultados: Los resultados muestran que aunque los materiales provistos por organizaciones no gubernamentales son más fáciles de leer, éstos están generalmente escritos a niveles de lectura más alta de la audiencia para la cual son desarrollados. Conclusiones: Se concluye que los materiales educativos de educación en salud para diabéticos no permiten una comprensión total de su contenido, ya que están escritos utilizando vocabulario más complejo que el que posee la población que los recibe.

Objective: To ascertain the health literacy levels of diabetes patient education materials distributed by government-funded or nonprofit organization. Methods: This descriptive study explored readability levels of 81 written diabetes health education materials available from healthcare providers. The Simplified Measure of Gobbledygoop (SMOG) readability formula and the Fray Graph were utilized to determine readability levels of diabetes patient information materials. The sample size excluded materials not in English and those with commercial purposes. Data analysis included measures of central tendency. In adition used t test and p-value. Results: Results from this study show that while education materials provided by nonprofit organizations are easier to read, they are still generally above the reading level of a large portion of the population they are intended to reach. Conclusion: Results from this study suggest that the majority of diabetes patient education materials are not adequate to reach their intended population due to high readability levels.

Niveles plasmáticos de mieloperoxidasa y proteína C reactiva en conejos machos en Nueva Zelanda expuestos a una dieta hiperlipidémica / Plasma concentration of myeloperoxidase and C reactive protein in male New Zealand rabbits, exposed to a hyperlipidemic diet

La Mieloperoxidasa (MPO) y la Proteína C Reactiva (PCR) han sido implicados en la fisiopatología de la aterosclerosis. El objetivo del presente estudio fue determinar las concentraciones plasmáticas de MPO y PCR y su relación con la formación de ateromas en conejos. Se estudiaron 23 conejos machos Nueva Zelanda: Grupo 1: conejarina y verdura; Grupo 2: Huevo y conejarina. El periodo experimental duró 13 semanas. Se determinó perfil lipídico por métodos enzimáticos, MPO por ELISA y PCR por turbidimetría en 0 13va semana. Se realizó estudio histológico de aorta. Los resultados revelaron que la PCR se elevó en el grupo 2 al final del estudio (p<0,05). No se observó diferencias en MPO en el grupo 2 en el estudio. En cuanto a los ateromas se evidenciaron lesiones tipo I y II en los conejos del grupo 2. En conclusión, se encontró que la PCR y no la MPO son marcadores de aterosclerosis según nuestras condiciones experimentales.

Myeloperoxidase (MPO) and C-reactive protein (CRP) have been implicated in atherosclerosis. The objective of the present study was to determine plasma concentration MPO and CRP and its relationship of formation of aortic lesions in rabbits. 23 male New Zealand rabbits were study: Group 1: “conejarina” (commercial rabbit food) and vegetables; Group 2: egg and conejarina. The experiment lasted 13 weeks. Lipid profile was done by enzymatic methods, MPO by ELISA, and PCR by turbidimetry in weeks 0 and 13. Histological study of rabbits’ aorta was done. Results revealed that in group 2 CRP increased at final study (p <0.05). No differences were observed in MPO values in the experiment. Regarding atheroma, group 2 presented type I and II lesions. In conclusion only CRP is marker of atherosclerosis according to our experimental conditions.

Ependymal cell differentiation and GLUT1 expression is a synchronous process in the ventricular wall.

Ependymal cells appear to be totally differentiated during the first 3 weeks in the mouse brain. Early during postnatal development ependymal cells differentiate and undergo metabolic activation, which is accompanied by increased glucose uptake. We propose that ependymal cells induce an overexpression of the glucose transporter, GLUT1, during the first 2 weeks after delivery in order to maintain the early metabolic activation. During the first postnatal day, GLUT1 is strongly induced in the upper region of the third ventricle and in the ventral area of the rostral cerebral aqueduct. During the next 4 days, GLUT1 is expressed in all differentiated ependymal cells of the third ventricle and in hypothalamic tanycytes. At the end of the first week, ependymal cell differentiation and GLUT1 overexpression is concentrated in the latero-ventral area of the aqueduct. We propose that ependymal cell differentiation and GLUT1 overexpression is a synchronous process in the ventricular wall.

Hypothalamic ependymal-glial cells express the glucose transporter GLUT2, a protein involved in glucose sensing.

The GLUT2 glucose transporter and the K-ATP-sensitive potassium channels have been implicated as an integral part of the glucose-sensing mechanism in the pancreatic islet beta cells. The expression of GLUT2 and K-ATP channels in the hypothalamic region suggest that they are also involved in a sensing mechanism in this area. The hypothalamic glial cells, known as tanycytes alpha and beta, are specialized ependymal cells that bridge the cerebrospinal fluid and the portal blood of the median eminence. We used immunocytochemistry, in situ hybridization and transport analyses to demonstrate the glucose transporters expressed in tanycytes. Confocal microscopy using specific antibodies against GLUT1 and GLUT2 indicated that both transporters are expressed in alpha and beta tanycytes. In addition, primary cultures of mouse hypothalamic tanycytes were found to express both GLUT1 and GLUT2 transporters. Transport studies, including 2-deoxy-glucose and fructose uptake in the presence or absence of inhibitors, indicated that these transporters are functional in cultured tanycytes. Finally, our analyses indicated that tanycytes express the K-ATP channel subunit Kir6.1 in vitro. As the expression of GLUT2 and K-ATP channel is linked to glucose-sensing mechanisms in pancreatic beta cells, we postulate that tanycytes may be responsible, at least in part, for a mechanism that allows the hypothalamus to detect changes in glucose concentrations.