Derivation of HVR1, HVR2 and HVR3 human embryonic stem cell lines from IVF embryos after preimplantation genetic diagnosis (PGD) for monogenic disorder

From 106 human blastocyts donate for research after in vitro fertilization (IVF) and preimplantation genetic diagnosis (PGD) for monogenetic disorder, 3 human embryonic stem cells (hESCs) HVR1, HVR2 and HVR3 were successfully derived. HVR1 was assumed to be genetically normal, HVR2 carrying Becker muscular dystrophy and HVR3 Hemophilia B. Despite the translocation t(9;15)(q34.3;q14) detected in HVR2, all the 3 cell lines were characterised in vitro and in vivo as normal hESCs lines and were registered in the Spanish Stem Cell Bank.Junta de Andalucía FEDER TCMR0021/06, PI246-2008Instituto de Salud Carlos III (FEDER) RD12/0019/0028, RD012/0036/0017, PI10/00964, PI11/02923, PI14/0101

Characterization of the "diabesity" gene HMG20A in pancreatic islets

Motivation: Type 2 Diabetes (T2D) accounts for 90-95% of diagnosed diabetic patients, which tendency in the next years is also expected to increase. Recent genomic wide association studies showed a correlation of an allelic variation of HMG20A with T2D in some ethnic groups. Up to date, there is no scientific evidence of the role of this gene in pancreatic tissue. But, in central nervous system, HMG20A regulates the expression of NeuroD, common in pancreas and nervous system morphogenesis. Here, our group makes an approach to characterize HMG20A in pancreatic islets, focusing on its involvement in glucose-stimulated insulin secretion (GSIS) and pancreatic islets development. We want to demonstrate that: 1) HMG20A is expressed in endocrine pancreas 2) HMG20A modifies expression of genes involved in pancreas development 3) silencing HMG20A affects expression levels of insulin secretion related genes and functionality.Methods: Qualitative expression of HMG20A is tested out in slides of pancreatic sections obtained from control mice. Co-localization with α or β cells is analyzed by immunofluorescence using anti-HMG20A, anti-insulin/glucagon antibodies and Dapi for nuclei. INS-1E cells are cultured and treated with a specific siRNA against HMG20A or a non-specific siRNA control during 72h. Genes involved in insulin secretion and endocrine pancreas development are assayed via qRT-PCR in INS1-E cells after siRNA treatment. Pdx1, Pax4, MafA and HMG20A expression levels are assessed following 2-ΔΔCt method. Finally, HMG20A silenced mouse islets and INS-1E are cultured at low glucose (2.8 mM) and high glucose medium (22 mM) following quantification of insulin secretion by ELISA.Results: Immunofluorescence confirmed co-localization of HMG20A with insulin (β-cell) and with glucagon (α-cells) producing cells in mouse pancreas. HMG20A expression diminished a 60% after treating INS1-E cells with a specific siRNA for HMG20A. Insulin secretion regulator gene, MafA, is downregulated significantly (50-60%) after HMG20A silencing. Pax4 expression significantly increased meanwhile Pdx1 showed a tendency to decrease. A 40% drop in insulin secretion is obtained in siHMG20A treated mouse islets compared to control.Conclusions: This data confirms HMG20A expression in pancreatic islets and impairment of insulin secretion when it is knocked down. Hence, concluding that HMG20A plays an important role in physiological GSIS and regulating pancreatic development related genes

Diferencias de género en las preferencias y desempeño profesional de ginecólogos y ginecólogas andaluces

En esta ponencia presentamos resultados parciales de una investigación desarrollada en Andalucía a una muestra representativa de profesionales de la ginecología que trabajan en el Sistema Sanitario Público Andaluz. El objetivo era conocer las opiniones y expectativas de los profesionales con respecto a la participación en actividades científicas y en el desempeño profesional. Los resultados muestran una fuerte incorporación de las mujeres al ejercicio de la Obstetricia y Ginecología y una desproporción en la ocupación de puestos de responsabilidad. El análisis desde la perspectiva de género y la teoría del gusto pone en relación las preferencias y el desempeño profesional con la persistencia de estereotipos y fronteras de género que se manifiestan en la dificultad que muestran las mujeres de participar como ponentes en reuniones científicas. Y muestra la dificultad de sacar conclusiones relativas a las diferencias observadas entre varones y mujeres si no se tiene en cuenta la variable edad

PAX4 preserves endoplasmic reticulum integrity preventing beta cell degeneration in a mouse model of type 1 diabetes mellitus

[Aims/hypothesis]: A strategy to enhance pancreatic islet functional beta cell mass (BCM) while restraining inflammation, through the manipulation of molecular and cellular targets, would provide a means to counteract the deteriorating glycaemic control associated with diabetes mellitus. The aims of the current study were to investigate the therapeutic potential of such a target, the islet-enriched and diabetes-linked transcription factor paired box 4 (PAX4), to restrain experimental autoimmune diabetes (EAD) in the RIP-B7.1 mouse model background and to characterise putative cellular mechanisms associated with preserved BCM. [Methods]: Two groups of RIP-B7.1 mice were genetically engineered to: (1) conditionally express either PAX4 (BPTL) or its diabetes-linked mutant variant R129W (mutBPTL) using doxycycline (DOX); and (2) constitutively express luciferase in beta cells through the use of RIP. Mice were treated or not with DOX, and EAD was induced by immunisation with a murine preproinsulin II cDNA expression plasmid. The development of hyperglycaemia was monitored for up to 4 weeks following immunisation and alterations in the BCM were assessed weekly by non-invasive in vivo bioluminescence intensity (BLI). In parallel, BCM, islet cell proliferation and apoptosis were evaluated by immunocytochemistry. Alterations in PAX4- and PAX4R129W-mediated islet gene expression were investigated by microarray profiling. PAX4 preservation of endoplasmic reticulum (ER) homeostasis was assessed using thapsigargin, electron microscopy and intracellular calcium measurements. [Results]: PAX4 overexpression blunted EAD, whereas the diabetes-linked mutant variant PAX4R129W did not convey protection. PAX4-expressing islets exhibited reduced insulitis and decreased beta cell apoptosis, correlating with diminished DNA damage and increased islet cell proliferation. Microarray profiling revealed that PAX4 but not PAX4R129W targeted expression of genes implicated in cell cycle and ER homeostasis. Consistent with the latter, islets overexpressing PAX4 were protected against thapsigargin-mediated ER-stress-related apoptosis. Luminal swelling associated with ER stress induced by thapsigargin was rescued in PAX4-overexpressing beta cells, correlating with preserved cytosolic calcium oscillations in response to glucose. In contrast, RNA interference mediated repression of PAX4-sensitised MIN6 cells to thapsigargin cell death. [Conclusions/interpretation]: The coordinated regulation of distinct cellular pathways particularly related to ER homeostasis by PAX4 not achieved by the mutant variant PAX4R129W alleviates beta cell degeneration and protects against diabetes mellitus. The raw data for the RNA microarray described herein are accessible in the Gene Expression Omnibus database under accession number GSE62846

Matemáticos y Matemáticas para el tercer milenio: de la abstracción a la realidad

Actas del congreso celebrado en San Fernando los días 7, 8, 9 y 10 de septiembre de 200

Fístula traqueo-esofágica en un perro como complicación de un cuerpo extraño esofágico y su resolución

En este artículo se describe un caso clínico de una fístula traqueo-esofágica en un perro mestizo de 3 años, como complicación de la extracción de un cuerpo extraño alojado en el esófago, así como su posterior resolución. El animal se presentó en nuestra consulta con una historia de regrugitación desde hacía una semana. En el estudio radiológico simple de la región cervical se pudo observar la presencia de una estructura irregular, de opacidad tejido ósea, compatible con un cuerpo extraño esofágico. La extracción se realizó mediante endoscopia, y tras ella se produjo, como consecuencia de la existencia de una fístula traqueo-esofágica, un enfisema subcutáneo en el área ventral del cuello. El tratamiento de esta complicación consistió en suturar la perforación traqueal y, debido a las amplias zonas de esófago desvitalizado, se optó por realizar una esofagectomía parcial, con miotomía del esófago y con refuerzo del área de anastomosis con un pedículo muscular del músculo esternotiroideo. Se practicó una gastrotomía de alimentación durante 10 días para proporcionar al animal soporte nutricional, y tras este periodo de timepo se realizó un esofagograma observándose una trayectoria normal del contraste. El animal evoluciono favorablemente aunque tiene que tomar una dieta blanda, y tras un años no se ha evidenciado ninguna complicación.

New applications for known drugs: Human glycogen synthase kinase 3 inhibitors as modulators of Aspergillus fumigatus growth [Postprint]

Invasive aspergillosis (IA) is one of the most severe forms of fungi infection. IA disease is mainly due to Aspergillus fumigatus, an air-borne opportunistic pathogen. Mortality rate caused by IA is still very high (50-95%), because of difficulty in early diagnostics and reduced antifungal treatment options, thus new and efficient drugs are necessary. The aim of this work is, using Aspergillus nidulans as non-pathogen model, to develop efficient drugs to treat IA. The recent discovered role of glycogen synthase kinase-3 homologue, GskA, in A. fumigatus human infection and our previous experience on human GSK-3 inhibitors focus our attention on this kinase as a target for the development of antifungal drugs. With the aim to identify effective inhibitors of colonial growth of A. fumigatus we use A. nidulans as an accurate model for in vivo and in silico studies. Several well-known human GSK-3β inhibitors were tested for inhibition of A. nidulans colony growth. Computational tools as docking studies and binding site prediction was used to explain the different biological profile of the tested inhibitors. Three of the five tested hGSK3β inhibitors are able to reduce completely the colonial growth by covalent bind to the enzyme. Therefore these compounds may be useful in different applications to eradicate IA.SAF2012-37979-C03-01 to A.M; BFU2012-33142 to E.A.EN

Incidencia de movimientos excitatorios tras el empleo de prpofol en la especie canina

El propofol es un anestésico intravenoso comúnmente usado en anestesia ya que proporciona una inducción suave y rápida, así como una recuperación corta y normalmente carente de efectos secundarios. Sin embargo, al igual que otros anestésicos, el propofol puede producir movimientos excitatorios tras su administración. En el presente estudio se revian un total de 70 inducciones anestésicas realizadas con propofol en cánidos sometidos a diferentes intervenciones quirúrgicas. La incidencia encontrada de movimientos excitatorios en estos casos fue del 8, 57%. Por tanto, es importante el reconocimiento de estos fenómenos, con el objetivo de proporcionar un tratamiento rápido y efectivo