Sonic hedgehog is basolaterally sorted from the TGN and transcytosed to the apical domain involving Dispatched-1 at Rab11-ARE.

Hedgehog proteins (Hhs) secretion from apical and/or basolateral domains occurs in different epithelial cells impacting development and tissue homeostasis. Palmitoylation and cholesteroylation attach Hhs to membranes, and Dispatched-1 (Disp-1) promotes their release. How these lipidated proteins are handled by the complex secretory and endocytic pathways of polarized epithelial cells remains unknown. We show that polarized Madin-Darby canine kidney cells address newly synthesized sonic hedgehog (Shh) from the TGN to the basolateral cell surface and then to the apical domain through a transcytosis pathway that includes Rab11-apical recycling endosomes (Rab11-ARE). Both palmitoylation and cholesteroylation contribute to this sorting behavior, otherwise Shh lacking these lipid modifications is secreted unpolarized. Disp-1 mediates first basolateral secretion from the TGN and then transcytosis from Rab11-ARE. At the steady state, Shh predominates apically and can be basolaterally transcytosed. This Shh trafficking provides several steps for regulation and variation in different epithelia, subordinating the apical to the basolateral secretion.

Participation of OCRL1, and APPL1, in the expression, proteolysis, phosphorylation and endosomal trafficking of megalin: Implications for Lowe Syndrome.

Megalin/LRP2 is the primary multiligand receptor for the re-absorption of low molecular weight proteins in the proximal renal tubule. Its function is significantly dependent on its endosomal trafficking. Megalin recycling from endosomal compartments is altered in an X-linked disease called Lowe Syndrome (LS), caused by mutations in the gene encoding for the phosphatidylinositol 5-phosphatase OCRL1. LS patients show increased low-molecular-weight proteins with reduced levels of megalin ectodomain in the urine and accumulation of the receptor in endosomal compartments of the proximal tubule cells. To gain insight into the deregulation of megalin in the LS condition, we silenced OCRL1 in different cell lines to evaluate megalin expression finding that it is post-transcriptionally regulated. As an indication of megalin proteolysis, we detect the ectodomain of the receptor in the culture media. Remarkably, in OCRL1 silenced cells, megalin ectodomain secretion appeared significantly reduced, according to the observation in the urine of LS patients. Besides, the silencing of APPL1, a Rab5 effector associated with OCRL1 in endocytic vesicles, also reduced the presence of megalin's ectodomain in the culture media. In both silencing conditions, megalin cell surface levels were significantly decreased. Considering that GSK3ß-mediated megalin phosphorylation reduces receptor recycling, we determined that the endosomal distribution of megalin depends on its phosphorylation status and OCRL1 function. As a physiologic regulator of GSK3ß, we focused on insulin signaling that reduces kinase activity. Accordingly, megalin phosphorylation was significantly reduced by insulin in wild-type cells. Moreover, even though in cells with low activity of OCRL1 the insulin response was reduced, the phosphorylation of megalin was significantly decreased and the receptor at the cell surface increased, suggesting a protective role of insulin in a LS cellular model.

Factores de riesgo de enfermedades prevalentes de la infancia en hogares de atención integral

RESUMEN Introducción: La Organización Mundial de la Salud manifiesta que las enfermedades prevalentes de la infancia en el 2016 causaron una mortalidad de 5 millones en menores de 5 años, a causa de neumonía y enfermedad diarreica aguda. Objetivo: Examinar los factores de riesgo modificables de enfermedades prevalentes de la infancia en los hogares comunitarios tradicionales del bienestar en Cúcuta, Colombia. Métodos: Se realizó un estudio observacional, descriptivo, transversal en el 2018, en 23 hogares del Instituto Colombiano del Bienestar Familiar en Cúcuta; la información se recolectó a partir del instrumento de identificación de factores de riesgo de enfermedades prevalentes de la infancia previo prueba inicial. Resultados: Se atendieron 276 menores, 56,8 % niños. Se identificaron factores de riesgo modificables en los hogares, en 22,0 % de estos, los infantes deben mejorar el lavado de manos; con un porcentaje similar, se demostró que se debe insistir en la limpieza del material didáctico, juguetes, ventiladores y cortinas; en 17,3 % se observaron condiciones regulares en cuanto a la infraestructura; además, 20,0 % de los menores compartían alimentos, se presentó asistencia de infantes con enfermedades en curso y ausencia de protocolos de atención a seguir en tales casos, entre otros. Conclusiones: Los factores de riesgo pueden ser intervenidos a través de la estrategia de atención integrada a las enfermedades prevalentes de la infancia desde sus componentes clínico, comunitario y organización local para su prevención, con el interés y comprensión de la responsabilidad compartida para la atención de esta problemática en salud.

ABSTRACT Introduction: The World Health Organization states that prevalent childhood diseases in 2016 caused a mortality of 5 million in children under 5 years old, due to pneumonia and acute diarrheal disease. Objective: Examine modifiable risk factors for prevalent childhood diseases in traditional community welfare homes in Cucuta, Colombia. Methods: An observational, descriptive, cross-sectional study was conducted in 2018 in 23 households of the Colombian Institute of Family Welfare in Cúcuta; the information was collected from the instrument to identify risk factors for prevalent childhood diseases after initial testing. Results: 276 children were attended, 56.8% of them were boys. Modifiable risk factors were identified in homes; in 22,0% of these, children should improve hand washing; with a similar percentage, it was shown that the cleaning of teaching materials, toys, fans and curtains should be insisted upon; in 17.3% regular infrastructure conditions were observed. In addition, 20,0% of the children shared food, there were children with ongoing diseases and absence of care protocols to follow in such cases, among others. Conclusions: The risk factors can be intervened through the strategy of integrated care for prevalent childhood diseases from its clinical, community and local organization components for its prevention, with the interest and understanding of the shared responsibility for the care of this health problem.

D-Propranolol Impairs EGFR Trafficking and Destabilizes Mutant p53 Counteracting AKT Signaling and Tumor Malignancy.

Cancer therapy may be improved by the simultaneous interference of two or more oncogenic pathways contributing to tumor progression and aggressiveness, such as EGFR and p53. Tumor cells expressing gain-of-function (GOF) mutants of p53 (mutp53) are usually resistant to EGFR inhibitors and display invasive migration and AKT-mediated survival associated with enhanced EGFR recycling. D-Propranolol (D-Prop), the non-beta blocker enantiomer of propranolol, was previously shown to induce EGFR internalization through a PKA inhibitory pathway that blocks the recycling of the receptor. Here, we first show that D-Prop decreases the levels of EGFR at the surface of GOF mutp53 cells, relocating the receptor towards recycling endosomes, both in the absence of ligand and during stimulation with high concentrations of EGF or TGF-α. D-Prop also inactivates AKT signaling and reduces the invasive migration and viability of these mutp53 cells. Unexpectedly, mutp53 protein, which is stabilized by interaction with the chaperone HSP90 and mediates cell oncogenic addiction, becomes destabilized after D-Prop treatment. HSP90 phosphorylation by PKA and its interaction with mutp53 are decreased by D-Prop, releasing mutp53 towards proteasomal degradation. Furthermore, a single daily dose of D-Prop reproduces most of these effects in xenografts of aggressive gallbladder cancerous G-415 cells expressing GOF R282W mutp53, resulting in reduced tumor growth and extended mice survival. D-Prop then emerges as an old drug endowed with a novel therapeutic potential against EGFR- and mutp53-driven tumor traits that are common to a large variety of cancers.

Asociación entre nivel y dominios de actividad física en escolares de 9 a 12 años / Association between level and domains of physical activity in schoolchildren from 9 to 12 years

RESUMEN Objetivo Analizar la relación entre el nivel y los dominios de la actividad física en escolares de 9 a 12 años de instituciones educativas públicas del Municipio de Los Patios, Cúcuta, 2017, con base en la información obtenida a través del cuestionario Physical Activity Questionnaire for Children (PAQ-C). Métodos Estudio descriptivo, transversal y analítico. Se realizó un muestreo estratificado proporcional con una muestra de 777 escolares. Se firmaron consentimientos y asentimientos informados, luego se procedió a la aplicación del cuestionario PAQ-C. Resultados El 50,3% de los escolares son niñas; el 44% presenta un nivel moderadamente activo, el dominio "tiempo libre" tiene mayor grado de asociación con un coeficiente de correlación de 0,65 y es significativo a un nivel de 0,01, seguido del dominio "hogar" con un coeficiente de correlación de 0,617 y en el dominio "escolar", 0,502. Conclusiones Se determinó que el nivel de actividad física de los escolares es moderadamente activo y que decrece con la edad. Se puede afirmar que los niños son más activos que las niñas, además, existe una correlación directa entre los dominios y el nivel general de actividad física.(AU)

ABSTRACT Objective To analyze the relationship between the level and domains of physical activity in school children from 9 to 12 years of public educational institutions of the Municipality of Los Patios, Cucuta, 2017, based on the information obtained through the questionnaire PAQ-C (Physical Activity Questionnaire for children). Methods Descriptive, transversal and analytical study. With a sample of 777 schoolchildren, a proportional stratified sampling was carried out. Consent and informed consent were signed, then the PAQ-C questionnaire was applied. Results 50,3% of schoolchildren are girls; 44% present a moderately active level, the free time domain has a greater degree of association with a correlation coefficient of 0,65 and is significant at a level of 0,01, followed by the household domain with a correlation coefficient of 0,617 and the school role 0,502. Conclusions It was determined that the level of physical activity of school children is moderately active and that it decreases with age, we can say that boys are more active than girls, in addition, there is a direct correlation between the domains and the general level of activity physical.(AU)

Estructura de un programa de ejercicio físico dirigido a escolares / Structure of a physical exercise program directed at schoolchildren

RESUMEN Objetivo Realizar una revisión temática sobre programas de ejercicio físico dirigidos a niños en el contexto mundial. Métodos La búsqueda de información se realizó en bases de datos Redalyc, Scielo, Elsevier, Pubmed, Scopus, utilizando los descriptores en salud como actividad motora, niños, adolescentes, ejercicio de calentamiento, ejercicios de estiramiento muscular y ejercicio de enfriamiento. Se seleccionaron artículos completos en idioma español e inglés publicados entre 2008 y 2019. Además, se consultaron páginas de organizaciones como la Organización Mundial de Salud (OMS), Organización Panamericana de la Salud (OPS), Ministerio de Salud y Protección Social de Colombia, el Departamento administrativo del deporte, la recreación, la actividad física y el aprovechamiento del tiempo libre, entre otros. Se organizó la información en base de datos Excel, acorde con el año de publicación, título, autores, metodología, fases y dimensiones del programa. Resultados Se analizó la información y se diseñó la estructura del programa de actividad dirigido a escolares, teniendo en cuenta las recomendaciones de la OMS y la legislación vigente en Colombia. Conclusiones La implementación de programas de ejercicio físico desde la infancia y la adolescencia promueve hábitos saludables que impactan de forma positiva la salud disminuyendo factores de riesgo de enfermedades crónicas no trasmisibles. La estructura de los mismos incluye características metodológicas como duración, frecuencia, intensidad, tipo de ejercicio y las fases de calentamiento, estiramiento, fase activa o central, y enfriamiento o vuelta a la calma.(AU)

ABSTRACT Objective To conduct a thematic review of physical exercise programs for children in the global context. Methods The search for information was carried out in databases Redalyc, Scielo, Elsevier, Pubmed, Scopus, using descriptors in health such as motor activity, children, adolescents, warm-up exercise, muscle stretching exercises and cooling exercise. Full-length articles in Spanish and English were selected and published between 2008 and 2019. In addition, pages were consulted from organizations such as the World Health Organization (WHO), Pan American Health Organization (PAHO), Ministry of Health and Social Protection of Colombia, the Administrative Department of Sports, recreation, physical activity and the use of free time, among others. The information was organized in an Excel database according to the year of publication, title, authors, methodology, phases and dimensions of the program. Results The information was analyzed and the structure of the activity program for schoolchildren was designed, taking into account the recommendations of the WHO and the legislation in force in Colombia. Conclusions The implementation of physical exercise programs from childhood and adolescence, promote healthy habits that positively impact health by decreasing risk factors of chronic non transmissible diseases. Their structure includes methodological characteristics such as duration, frequency, intensity, type of exercise and the warm-up, stretching, active or central phase, and cooling down or returning to calm phases.(AU)

[Association between level and domains of physical activity in schoolchildren from 9 to 12 years]. / Asociación entre nivel y dominios de actividad física en escolares de 9 a 12 años.

OBJECTIVE: To analyze the relationship between the level and domains of physical activity in school children from 9 to 12 years of public educational institutions of the Municipality of Los Patios, Cucuta, 2017, based on the information obtained through the questionnaire PAQ-C (Physical Activity Questionnaire for children). METHODS: Descriptive, transversal and analytical study. With a sample of 777 schoolchildren, a proportional stratified sampling was carried out. Consent and informed consent were signed, then the PAQ-C questionnaire was applied. RESULTS: 50,3% of schoolchildren are girls; 44% present a moderately active level, the free time domain has a greater degree of association with a correlation coefficient of 0,65 and is significant at a level of 0,01, followed by the household domain with a correlation coefficient of 0,617 and the school role 0,502. CONCLUSIONS: It was determined that the level of physical activity of school children is moderately active and that it decreases with age, we can say that boys are more active than girls, in addition, there is a direct correlation between the domains and the general level of activity physical.

OBJETIVO: Analizar la relación entre el nivel y los dominios de la actividad física en escolares de 9 a 12 años de instituciones educativas públicas del Municipio de Los Patios, Cúcuta, 2017, con base en la información obtenida a través del cuestionario Physical Activity Questionnaire for Children (PAQ-C). MÉTODOS: Estudio descriptivo, transversal y analítico. Se realizó un muestreo estratificado proporcional con una muestra de 777 escolares. Se firmaron consentimientos y asentimientos informados, luego se procedió a la aplicación del cuestionario PAQ-C. RESULTADOS: El 50,3% de los escolares son niñas; el 44% presenta un nivel moderadamente activo, el dominio "tiempo libre" tiene mayor grado de asociación con un coeficiente de correlación de 0,65 y es significativo a un nivel de 0,01, seguido del dominio "hogar" con un coeficiente de correlación de 0,617 y en el dominio "escolar", 0,502. CONCLUSIONES: Se determinó que el nivel de actividad física de los escolares es moderadamente activo y que decrece con la edad. Se puede afirmar que los niños son más activos que las niñas, además, existe una correlación directa entre los dominios y el nivel general de actividad física.

[Structure of a physical exercise program directed at schoolchildren]. / Estructura de un programa de ejercicio físico dirigido a escolares.

OBJECTIVE: To conduct a thematic review of physical exercise programs for children in the global context. METHODS: The search for information was carried out in databases Redalyc, Scielo, Elsevier, Pubmed, Scopus, using descriptors in health such as motor activity, children, adolescents, warm-up exercise, muscle stretching exercises and cooling exercise. Full-length articles in Spanish and English were selected and published between 2008 and 2019. In addition, pages were consulted from organizations such as the World Health Organization (WHO), Pan American Health Organization (PAHO), Ministry of Health and Social Protection of Colombia, the Administrative Department of Sports, recreation, physical activity and the use of free time, among others. The information was organized in an Excel database according to the year of publication, title, authors, methodology, phases and dimensions of the program. RESULTS: The information was analyzed and the structure of the activity program for schoolchildren was designed, taking into account the recommendations of the WHO and the legislation in force in Colombia. CONCLUSIONS: The implementation of physical exercise programs from childhood and adolescence, promote healthy habits that positively impact health by decreasing risk factors of chronic non transmissible diseases. Their structure includes methodological characteristics such as duration, frequency, intensity, type of exercise and the warm-up, stretching, active or central phase, and cooling down or returning to calm phases.

OBJETIVO: Realizar una revisión temática sobre programas de ejercicio físico dirigidos a niños en el contexto mundial. MÉTODOS: La búsqueda de información se realizó en bases de datos Redalyc, Scielo, Elsevier, Pubmed, Scopus, utilizando los descriptores en salud como actividad motora, niños, adolescentes, ejercicio de calentamiento, ejercicios de estiramiento muscular y ejercicio de enfriamiento. Se seleccionaron artículos completos en idioma español e inglés publicados entre 2008 y 2019. Además, se consultaron páginas de organizaciones como la Organización Mundial de Salud (OMS), Organización Panamericana de la Salud (OPS), Ministerio de Salud y Protección Social de Colombia, el Departamento administrativo del deporte, la recreación, la actividad física y el aprovechamiento del tiempo libre, entre otros. Se organizó la información en base de datos Excel, acorde con el año de publicación, título, autores, metodología, fases y dimensiones del programa. RESULTADOS: Se analizó la información y se diseñó la estructura del programa de actividad dirigido a escolares, teniendo en cuenta las recomendaciones de la OMS y la legislación vigente en Colombia. CONCLUSIONES: La implementación de programas de ejercicio físico desde la infancia y la adolescencia promueve hábitos saludables que impactan de forma positiva la salud disminuyendo factores de riesgo de enfermedades crónicas no trasmisibles. La estructura de los mismos incluye características metodológicas como duración, frecuencia, intensidad, tipo de ejercicio y las fases de calentamiento, estiramiento, fase activa o central, y enfriamiento o vuelta a la calma.

OCRL controls trafficking through early endosomes via PtdIns4,5P2-dependent regulation of endosomal actin.

Mutations in the phosphatidylinositol 4,5-bisphosphate (PtdIns4,5P(2)) 5-phosphatase OCRL cause Lowe syndrome, which is characterised by congenital cataracts, central hypotonia, and renal proximal tubular dysfunction. Previous studies have shown that OCRL interacts with components of the endosomal machinery; however, its role in endocytosis, and thus the pathogenic mechanisms of Lowe syndrome, have remained elusive. Here, we show that via its 5-phosphatase activity, OCRL controls early endosome (EE) function. OCRL depletion impairs the recycling of multiple classes of receptors, including megalin (which mediates protein reabsorption in the kidney) that are retained in engorged EEs. These trafficking defects are caused by ectopic accumulation of PtdIns4,5P(2) in EEs, which in turn induces an N-WASP-dependent increase in endosomal F-actin. Our data provide a molecular explanation for renal proximal tubular dysfunction in Lowe syndrome and highlight that tight control of PtdIns4,5P(2) and F-actin at the EEs is essential for exporting cargoes that transit this compartment.

Marlin-1 and conventional kinesin link GABAB receptors to the cytoskeleton and regulate receptor transport.

The cytoskeleton and cytoskeletal motors play a fundamental role in neurotransmitter receptor trafficking, but proteins that link GABA(B) receptors (GABA(B)Rs) to the cytoskeleton have not been described. We recently identified Marlin-1, a protein that interacts with GABA(B)R1. Here, we explore the association of GABA(B)Rs and Marlin-1 to the cytoskeleton using a combination of biochemistry, microscopy and live cell imaging. Our results indicate that Marlin-1 is associated to microtubules and the molecular motor kinesin-I. We demonstrate that a fraction of Marlin-1 is mobile in dendrites of cultured hippocampal neurons and that mobility is microtubule-dependent. We also show that GABA(B)Rs interact robustly with kinesin-I and that intracellular membranes containing GABA(B)Rs are sensitive to treatments that disrupt a protein complex containing Marlin-1, kinesin-I and tubulin. Finally, we report that a kinesin-I mutant severely impairs receptor transport. We conclude that Marlin-1 and kinesin-1 link GABA(B)Rs to the tubulin cytoskeleton in neurons.