Cinco años del programa estímulo a la investigación “Lisandro Alvarado” (PEILA): respuesta institucional a la labor del investigador de la UCLA

The University Council of the Centroccidental "Lisandro Alvarado" University (UC-UCLA) on 27-07-2011 approved the request of the Scientific Humanistic and Technological Development Council (CDCHT) for the creation of the "Lisandro Alvarado" Stimulus to Research Program (PEILA). It was implemented as of 2012. During these five years this Program has recognized the work of UCLA’s researchers. The PEILA, based on exclusively academic criteria, classifies researchers into three levels, which receive resources to support their research activity. Accredited researchers are recognized annually for their publications with an additional bonus for them. The program started with 249 accredited researchers in seven areas of knowledge and by 2016 were 161 registered and although level I decreased 43%, levels II and III increased 19 and 3% respectively. The results for the period 2012-2016 show that, despite the budget crisis, the PEILA has been consolidated as an instrument for institutional recognition and stimulation of university research at UCLAEl Consejo Universitario de la Universidad Centroccidental “Lisandro Alvarado” (CU-UCLA) el 27-07-2011 aprobó la solicitud del Consejo de Desarrollo Científico Humanístico y Tecnológico (CDCHT) para la creación del Programa de Estímulo a la Investigación “Lisandro Alvarado” (PEILA). El mismo fue implementado a partir del 2012. Durante estos cinco años este Programa ha reconocido la labor del investigador de la UCLA. El PEILA con base en criterios exclusivamente académicos clasifica a los acreditados en tres niveles, los cuales reciben recursos para apoyar su actividad investigativa. Actualmente los investigadores acreditados reciben un incentivo en forma de un fondo de trabajo académico sujeto a rendición de cuentas y adicionalmente les son reconocidas anualmente sus publicaciones con una bonificación sin rendición de cuentas. El programa se inicia con 249 investigadores acreditados en siete áreas del saber y para 2016 había 161 registrados y aunque el nivel I disminuyó en 43%, los niveles II y III aumentaron en 19 y 3% respectivamente. Los resultados durante el período 2012-2016 demuestran que a pesar de la crisis presupuestaria, el PEILA se ha consolidado como instrumento de reconocimiento institucional y estímulo a la investigación universitaria en la UCL

Grado de retinopatía hipertensiva según la escala de Keith-Wagener-Baker en mujeres preeclámpticas entre 18 y 35 años estudio multicéntrico en los hospitales públicos de la ciudad de Quito de julio a diciembre de 2016

La preeclampsia es una patología característica de la gestación, alumbramiento y puerperio, observada a partir de las 20 semanas de gestación hasta las 12 semanas del puerperio, considerada como una de las causas más importantes de morbimortalidad materna a nivel mundial. La vía visual se ve afectada en diversas maneras como: empeoramiento de la retinopatía diabética, retinopatía serosa central, retinopatía hipertensiva, oclusión vascular. Métodos: El presente estudio propuso clasificar la presencia de Retinopatía Hipertensiva valorada con fondo de ojo mediante Oftalmoscopia Directa en gestantes con preeclampsia leve y severa en los servicios de Obstetricia de los Hospitales Públicos de Quito entre julio a diciembre del 2016. Se realizó oftalmoscopia directa mediante el dispositivo iExaminer (iPhone 4) de que convierte el oftalmoscopio PanOptic (Welch Allyn) en un dispositivo de imagen digital móvil. Fueron evaluadas un total de 96 pacientes con diagnóstico de preeclampsia leve y severa. Resultados: Se diagnosticó retinopatía grado I en 44,79% de las pacientes. El 17,70% de pacientes con retinopatía grado I tenían edades comprendidas entre 31 a 35 años (X2=0,047). El 30,20% de las pacientes desarrollaron retinopatía grado I en el contexto de preeclampsia severa. Los escotomas se presentaron en 28,12% y ambliopía en 9,37% de pacientes con retinopatía grado I. Desarrollaron Preeclampsia Leve el 40,62% encontrándose entre 37 y 41 semanas de edad gestacional mientras 26,04% presentó Preeclampsia severa entre las semanas 37 a 41 (X2=0,010). Conclusión: En este estudio se evidenció correlación significativa entre retinopatía y la edad de las pacientes así como con edad gestacional. La retinopatía por sí sola, no puede predecir el pronóstico materno en las pacientes con preeclampsi

Predicción computacional de la estructura terciaria de la iduronato 2-sulfato sulfatasa humana

Introduction. Hunter syndrome (MC KUSIK 309900) or mucopolysacharidosis type II is due to the deficiency of the enzyme iduronate 2 sulfate sulfatase (E.C. 3.1.6.13). This enzyme has not been crystallized, and therefore the experimental structures are not available. Objectives. A computational three-dimensional model was proposed for the iduronate 2 sulfate sulfatase enzyme.Materials and methods. A computational analysis of this enzyme used the following free internet software programs: Comput pI/MW, JaMBW Chapter 3.1.7, SWISS-MODEL, Geno3d, ProSup. Energy minimization was done with Discover 3 and Insight II version 2004.Results. A three-dimensional conformational model was proposed. The model showed 33.3% of helix structure, 7.2% beta sheet, and 59.5% random coil. RMS values (Root Mean Square) (0.78 and 0.86Å) were found when compared with other enzymes of the same family. The model presented 5 exposed N-glycosylation potential sites and an entry to the pocket that contains the amino acids of the active site. A high correlation was found between the type of mutations and the severity of the phenotype in twenty patients analyzed.Conclusion. The RMS values, as well as the high correlation between the type of mutation and the phenotype, indicated that the model predicts some aspects of the enzyme's biological behavior.Introducción. El síndrome de Hunter o mucopolisacaridosis tipo II (MC KUSIK 309900) es causado por la deficiencia de la iduronato 2-sulfato sulfatasa humana (E.C. 3.1.6.13). La enzima no ha sido cristalizada y por tanto sus estructuras no se conocen por deducción experimental.Objetivo. Proponer un modelo computacional para la estructura tridimensional de la iduronato 2-sulfato sulfatasa humana.Materiales y métodos. Se realizó un análisis computacional de esta enzima empleando programas de libre acceso en internet como Comput pI/MW, JaMBW Chapter 3.1.7, SWISSMODEL, 3D-PSSM, ProSup. Los procesos de minimización de energía se realizaron con el programa Discover 3 del paquete Insight II (2004).Resultados. Se propone un modelo tridimensional de la iduronato 2-sulfato sulfatasa humana que presenta 33,3% en hélice, 7,2% en hoja plegada y 59,5% en enrollamiento al azar (coil). Se hallaron valores de RMS (del inglés Root Mean Square) de 0,78 y 0,86Å al compararla con otras enzimas de la misma familia. El modelo revela cinco sitios potenciales de N-glicosilación y una entrada al bolsillo que contiene los aminoácidos que componen el sitio activo. Usando este modelo se encontró una buena correlación entre el tipo de mutaciones y la gravedad de la enfermedad en 20 pacientes analizados.Conclusión. Los valores de RMS y la correlación genotipo-fenotipo en los pacientes analizados sugieren el modelo puede usarse para predecir ciertos aspectos del comportamiento biológico de la enzima

Low-Scale Expression and Purification of an Active Putative Iduronate 2-Sulfate Sulfatase-Like Enzyme from Escherichia coli K12

The sulfatase family involves a group of enzymes with a large degree of similarity. Until now, sixteen human sulfatases have been identified, most of them found in lysosomes. Human deficiency of sulfatases generates various genetic disorders characterized by abnormal accumulation of sulfated intermediate compounds. Mucopolysaccharidosis type II is characterized by the deficiency of iduronate 2-sulfate sulfatase (IDS), causing the lysosomal accumulation of heparan and dermatan sulfates. Currently, there are several cases of genetic diseases treated with enzyme replacement therapy, which have generated a great interest in the development of systems for recombinant protein expression. In this work we expressed the human recombinant IDS-Like enzyme (hrIDS-Like) in Escherichia coli DH5α. The enzyme concentration revealed by ELISA varied from 78. 13 to 94. 35 ng/ml and the specific activity varied from 34. 20 to 25. 97 nmol/h/mg. Western blotting done after affinity chromatography purification showed a single band of approximately 40 kDa, which was recognized by an IgY polyclonal antibody that was developed against the specific peptide of the native protein. Our 100 ml-shake-flask assays allowed us to improve the enzyme activity seven fold, compared to the E. coli JM109/pUC13-hrIDS-Like system. Additionally, the results obtained in the present study were equal to those obtained with the Pichia pastoris GS1115/pPIC-9-hrIDS-Like system (3 L bioreactor scale). The system used in this work (E. coli DH5α/pGEX-3X-hrIDS-Like) emerges as a strategy for improving protein expression and purification, aimed at recombinant protein chemical characterization, future laboratory assays for enzyme replacement therapy, and as new evidence of active putative sulfatase production in E. coli. © 2013 The Microbiological Society of Korea and Springer-Verlag Berlin Heidelberg.Fil: Morales Álvarez, Edwin David. Universidad del Quindio; ColombiaFil: Rivera Hoyos, Claudia Marcela. Universidad del Quindio. Facultad de Medicina; ColombiaFil: Baena Moncada, Angelica Maria. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Córdoba; ArgentinaFil: Landázuri, Patricia. Universidad del Quindio. Facultad de Medicina. Centro de Investig. Biomédicas; ColombiaFil: Poutou Piñales, Raúl A.. Pontificia Universidad Javeriana; ColombiaFil: Sáenz Suárez, Homero. Universidad del Quindio; ColombiaFil: Barrera, Luis A.. Pontificia Universidad Javeriana; ColombiaFil: Echeverri Peña, Olga Y.. Pontificia Universidad Javeriana; Colombi

LA TERAPIA DE REEMPLAZO ENZIMÁTICO EN EL TRATAMIENTO DE ENFERMEDADES GENÉTICAS

The aim of enzyme reemplacement therapy (ERT), is to provide a foreign protein to a patient who is producing it abnormally. The protein to be  used may be purified from human fluids or tissues and from bacteria, yeast or mammalian cells in which the corresponding human gene has been introduced. Up to now three diseases are currently being treated worldwide with ERT, Gaucher, Fabry and the combined severe human immunodeficiency. Advanced studies are in progress for the following diseases Hurler, Hunter, Pompe, Niemann-Pick B type, Morquio A,  hereditary angioderma and penfigo vulgar. One of the problems of using ERT is the its high costs, due to both the low number of patients affected world-wide and the very expensive procedures required to obtain protein expression in mammalian cells. To overcome these problems there are two possible alternatives: expression in bacteria or in yeast, however bacteria do not glycosylate proteins and most of inborn errors of  metabolism are caused by deficiency in glycosilated proteins (GP).  Yeast glycosilate proteins, but they have different pattern of   glycosylation from mammalia

LA TERAPIA DE REEMPLAZO ENZIMÁTICO EN EL TRATAMIENTO DE ENFERMEDADES GENÉTICAS

The aim of enzyme reemplacement therapy (ERT), is to provide a foreign protein to a patient who is producing it abnormally. The protein to be  used may be purified from human fluids or tissues and from bacteria, yeast or mammalian cells in which the corresponding human gene has been introduced. Up to now three diseases are currently being treated worldwide with ERT, Gaucher, Fabry and the combined severe human immunodeficiency. Advanced studies are in progress for the following diseases Hurler, Hunter, Pompe, Niemann-Pick B type, Morquio A,  hereditary angioderma and penfigo vulgar. One of the problems of using ERT is the its high costs, due to both the low number of patients affected world-wide and the very expensive procedures required to obtain protein expression in mammalian cells. To overcome these problems there are two possible alternatives: expression in bacteria or in yeast, however bacteria do not glycosylate proteins and most of inborn errors of  metabolism are caused by deficiency in glycosilated proteins (GP).  Yeast glycosilate proteins, but they have different pattern of   glycosylation from mammalia

Implicaciones de las proteínas de choque térmico (sHsp/HSPB) en el desarrollo de enfermedades degenerativas

The small heat shock proteins (sHsp/HSPB) and their implication in the development of degenerative diseases. Stress proteins are presentin all the cells and participate in the synthesis of proteins binding their selves to the newly formed peptides to direct their folding, thusensuring their three-dimensional structure and appropriate functioning. Besides, stress proteins are able to bind to damaged peptides andproteins due to diverse types of aggressions, enabling their repair or degradation. When cells are exposed to adverse situations, a rapidincrease in concentration of stress proteins occurs. Stress protein expression had been associated to heat shocks only, but nowadays weknow that stress proteins are induced as a response to a wide array of physiological and environmental aggressions such as: viral infections,inflammations, febrile responses, cell exposure to cytotoxic compounds, pH acidification, anoxia, and heat shock. The production of thistype of molecules is a defense mechanism that allows the cell to adapt to anomalous situations and increase its survival capacity. In our studywe present a brief historical account on stress proteins, their association with some pathologies, and discuss the current state of knowledgeabout this type of molecules and the possible mechanisms involved in protein conformational disorders or proteopathies

Implicaciones de las proteínas de choque térmico (sHsp/HSPB) en el desarrollo de enfermedades degenerativas

Las proteínas de choque térmico pertenecen al grupo de proteínas de estrés y son moléculas presentes en todas las células, se unen a los péptidos nacientes para dirigir su plegamiento,  garantizando su estructura tridimensional y con ello su funcionamiento correcto. Dentro de éste grupo de moléculas se encuentran las proteínas de choque térmico pequeñas (sHsp/HSPB), también,  capaces de unirse a péptidos y proteínas dañadas por diversos tipos de agresiones, facilitando su reparación o degradación. En células expuestas a situaciones adversas se produce un rápido aumento en las concentraciones de estas proteínas. Aunque inicialmente se asoció la expresión de las proteínas de estrés a aumentos bruscos de temperatura, hoy día es conocido que existen en condiciones  fisiológicas normales y sus concentraciones se aumentan en respuesta a un amplio espectro de agresiones ambientales como: infecciones virales, inflamaciones, cuadros febriles, exposición a compuestos citotóxicos, acidificación del pH, anoxia o shock térmico. La producción de este tipo de moléculas constituye un mecanismo de defensa que permite a la célula adaptarse a condiciones anómalas y aumentar su capacidad de supervivencia. Se presenta en este trabajo, una breve reseña histórica de las proteínas de choque térmico pequeñas, su asociación con algunas patologías y se discute el estado actual del conocimiento  de este tipo de moléculas y los posibles mecanismos implicados en las enfermedades conformacionales o proteinopatías, en donde existen alteraciones en la conformación nativa de las proteínas.Palabras clave: Proteínas de choque térmico pequeñas (sHsp/HSPB), cuerpos de inclusión, enfermedades degenerativas, miopatías, mutaciones. AbstractImplication of stress proteins in degenerative diseases. Stress proteins are present in all the cells and participate in the synthesis of proteins binding their selves to the newly formed peptides to direct their folding, thus ensuring their three-dimensional structure and appropriate functioning. Besides, stress proteins are able to bind to damaged peptides and proteins due to diverse types of aggressions, enabling their repair or degradation. When cells are exposed to adverse situations, a rapid increase in concentration of stress proteins occurs. Stress protein expression had been associated to heat shocks only, but nowadays we know that stress proteins are induced as a response to a wide array of physiological and environmental aggressions such as: viral infections, inflammations, febrile responses, cell exposure to cytotoxic compounds, pH acidification, anoxia, and heat shock. The production of this type of molecules is a defense mechanism that allows the cell to adapt to anomalous situations and increase its survival capacity. In our study we present a brief historical account on stress proteins, their association with some pathologies, and discuss the current state of knowledge about this type of molecules and the possible mechanisms involved in protein conformational disorders or proteopathies. Key words: Heat shock proteins (Hsp), body inclusion, degenerative diseases, myopathies, mutations.Resumo Envolvimentos das proteínas de estresse em doenças degenerativas. As proteínas do estresse são moléculas presentes em todas as células e estão envolvidas no processo de síntese protéica, ligando-se aos peptídeos nascentes para dirigir o seu dobramento, o que garante sua estrutura tridimensional e, consequentemente, o seu bom funcionamento. Além disso, são capazes de se ligar a peptídeos e proteínas danificadas por diferentes tipos de agressão, facilitando a sua reparação ou degradação. Quando as células ficam expostas a situações adversas aumentam rapidamente as concentrações dessas proteínas. Embora no inicio associou-se a expressão das proteínas de estresse com uma mudança brusca de temperatura, atualmente é conhecido que elas são induzidas como resposta a um amplo espectro de agressões fisiológicas e ambientais, tais como: infecções virais, inflamações, febres, exposição das células a compostos citotóxicos, a acidificação do pH, anoxia ou choque térmico. A produção de tais células é um mecanismo de defesa que permite à célula se adaptar a condições anormais e aumentar a sua capacidade de sobrevivência. Neste trabalho se apresenta um breve histórico das proteínas de estresse, a sua associação com certas doenças e se discute o estado atual do conhecimento sobre este tipo de moléculas e os possíveis mecanismos envolvidos nas doenças conformacionais ou proteinopatias. Palavras chave: proteínas de choque térmico (Hsp), corpos de inclusão, doenças degenerativas, miopatias, mutaçõe

Implicaciones de las proteínas de choque térmico (sHsp/HSPB) en el desarrollo de enfermedades degenerativas

Las proteínas de choque térmico pertenecen al grupo de proteínas de estrés y son moléculas presentes en todas las células, se unen a los péptidos nacientes para dirigir su plegamiento,  garantizando su estructura tridimensional y con ello su funcionamiento correcto. Dentro de éste grupo de moléculas se encuentran las proteínas de choque térmico pequeñas (sHsp/HSPB), también,  capaces de unirse a péptidos y proteínas dañadas por diversos tipos de agresiones, facilitando su reparación o degradación. En células expuestas a situaciones adversas se produce un rápido aumento en las concentraciones de estas proteínas. Aunque inicialmente se asoció la expresión de las proteínas de estrés a aumentos bruscos de temperatura, hoy día es conocido que existen en condiciones  fisiológicas normales y sus concentraciones se aumentan en respuesta a un amplio espectro de agresiones ambientales como: infecciones virales, inflamaciones, cuadros febriles, exposición a compuestos citotóxicos, acidificación del pH, anoxia o shock térmico. La producción de este tipo de moléculas constituye un mecanismo de defensa que permite a la célula adaptarse a condiciones anómalas y aumentar su capacidad de supervivencia. Se presenta en este trabajo, una breve reseña histórica de las proteínas de choque térmico pequeñas, su asociación con algunas patologías y se discute el estado actual del conocimiento  de este tipo de moléculas y los posibles mecanismos implicados en las enfermedades conformacionales o proteinopatías, en donde existen alteraciones en la conformación nativa de las proteínas.Palabras clave: Proteínas de choque térmico pequeñas (sHsp/HSPB), cuerpos de inclusión, enfermedades degenerativas, miopatías, mutaciones. AbstractImplication of stress proteins in degenerative diseases. Stress proteins are present in all the cells and participate in the synthesis of proteins binding their selves to the newly formed peptides to direct their folding, thus ensuring their three-dimensional structure and appropriate functioning. Besides, stress proteins are able to bind to damaged peptides and proteins due to diverse types of aggressions, enabling their repair or degradation. When cells are exposed to adverse situations, a rapid increase in concentration of stress proteins occurs. Stress protein expression had been associated to heat shocks only, but nowadays we know that stress proteins are induced as a response to a wide array of physiological and environmental aggressions such as: viral infections, inflammations, febrile responses, cell exposure to cytotoxic compounds, pH acidification, anoxia, and heat shock. The production of this type of molecules is a defense mechanism that allows the cell to adapt to anomalous situations and increase its survival capacity. In our study we present a brief historical account on stress proteins, their association with some pathologies, and discuss the current state of knowledge about this type of molecules and the possible mechanisms involved in protein conformational disorders or proteopathies. Key words: Heat shock proteins (Hsp), body inclusion, degenerative diseases, myopathies, mutations.Resumo Envolvimentos das proteínas de estresse em doenças degenerativas. As proteínas do estresse são moléculas presentes em todas as células e estão envolvidas no processo de síntese protéica, ligando-se aos peptídeos nascentes para dirigir o seu dobramento, o que garante sua estrutura tridimensional e, consequentemente, o seu bom funcionamento. Além disso, são capazes de se ligar a peptídeos e proteínas danificadas por diferentes tipos de agressão, facilitando a sua reparação ou degradação. Quando as células ficam expostas a situações adversas aumentam rapidamente as concentrações dessas proteínas. Embora no inicio associou-se a expressão das proteínas de estresse com uma mudança brusca de temperatura, atualmente é conhecido que elas são induzidas como resposta a um amplo espectro de agressões fisiológicas e ambientais, tais como: infecções virais, inflamações, febres, exposição das células a compostos citotóxicos, a acidificação do pH, anoxia ou choque térmico. A produção de tais células é um mecanismo de defesa que permite à célula se adaptar a condições anormais e aumentar a sua capacidade de sobrevivência. Neste trabalho se apresenta um breve histórico das proteínas de estresse, a sua associação com certas doenças e se discute o estado atual do conhecimento sobre este tipo de moléculas e os possíveis mecanismos envolvidos nas doenças conformacionais ou proteinopatias. Palavras chave: proteínas de choque térmico (Hsp), corpos de inclusão, doenças degenerativas, miopatias, mutaçõe