Método de detección de la Telangiectasia Hemorrágica Hereditaria

Método de detección de la Telangiectasia Hemorrágica Hereditaria. Método para la detección de la Telangiectasia Hemorrágica Hereditaria (HHT) que comprende el análisis in vitro de los productos de la expresión de los genes ENG o FLT1 Y ENG o ANGPT2 en muestras biológicas de pacientes y que, además, permite sub-clasificar las variantes HHT1 y HHT2 de la enfermedad. En particular, la presente invención se refiere a un kit para la detección molecular de HHT capaz de llevar a cabo la detección mencionada anteriormente.Peer reviewedConsejo Superior de Investigaciones Científicas (España), Centro de Investigación Biomédica en Red de Enfermedades Raras (CIBERER)A1 Solicitud de patentes con informe sobre el estado de la técnic

Biomarcadores plasmáticos y alteraciones de la respuesta inmunitaria en la telangiectasia hemorrágica hereditaria (HHT)

232 p.-64 fig.-17 tab.-anexos.La Telangiectasia Hemorrágica Hereditaria (HHT) también conocida como síndrome de Rendu-Osler-Weber es una enfermedad rara caracterizada por la presencia de paredes vasculares débiles que dan lugar a la aparición de malformaciones arteriovenosas (MAVs). Las MAVs pueden presentarse en piel y mucosas dando lugar a la aparición de Telangiectasias, y aquellas presentes en la mucosa nasal dan lugar a epistaxis (sangrados nasales recurrentes). Las MAVs de mayor tamaño pueden presentarse en órganos internos como pulmón, hígado y cerebro y su evolución puede desencadenar hemorragias internas que podrían tener consecuencias fatales para el paciente si no son localizadas precozmente y tratadas. En la actualidad, el diagnóstico de HHT se realiza en base a los Criterios clínicos de Curaçao, basados en la sintomatología clínica clásica HHT y teniendo en cuenta si existe un componente hereditario. Existen 5 tipos de HHT clasificadas en base al gen o locus afectado. En más del 90% de los casos las mutaciones están localizadas en ENG (gen endoglina) o ACVRL1 (gen ALK-1), que dan lugar a la HHT1 y HHT2, respectivamente. En el 10% de los casos restantes la mutación puede localizarse en un locus del cromosoma 5 (gen sin identificar), un locus del cromosoma 7 (gen sin identificar) o bien en GDF2 (gen BMP9). Finalmente, ha sido identificado un síndrome conocido como poliposis juvenial asociado a la HHT causado por mutaciones en MADH4 (gen Smad4). Todos los genes identificados codifican proteínas pertenecientes al sistema de señalización de TGF-β1. El diagnóstico HHT viene confirmado mediante el diagnóstico genético, pero éste puede prolongarse en el tiempo en especial cuando no existe un caso índice en la familia. Bien sea por el desconocimiento de la mutación familiar o porque los individuos no cumplen los criterios de Curaçao, frecuente en edad infantil o individuos jóvenes, un diagnóstico precoz alternativo al genético sería de gran utilidad para poder iniciar el screening del paciente lo antes posible y aplicar un tratamiento en el caso de que fuese necesario, traduciéndose ello en una mejora de la calidad de vida.Es por ello que uno de los primeros objetivos del presente trabajo fue hallar un mecanismo sencillo y rápido de realizar un diagnóstico a partir de muestras sanguíneas. En consecuencia, hemos establecido una fórmula diagnóstica mediante la valoración de los niveles de endoglina soluble (sEng) y angiopoietina-2 (Ang-2), que nos permite identificar a los individuos afectos de HHT1 o HHT2 con una elevada fiabilidad y rápidamente puesto que en 6 horas se pueden realizar las valoraciones mediante técnicas de ELISA. Como se ha mencionado anteriormente, la HHT se caracteriza por la presencia de MAVs cuya localización y/o el resultado de hemorragias derivadas de las mismas, dan lugar a los síntomas clásicos que constituyen a su vez los criterios clínicos de Curaçao. Si bien, en los últimos años son varios los estudios y publicaciones que sugieren que la HHT podría estar relacionada con una deficiencia en la respuesta inmunitaria. Algunas publicaciones muestran que hay una mayor incidencia y recurrencia de enfermedades infecciosas en pacientes HHT. Se han descrito abscesos cerebrales, osteomielitis, espondilodiscitis y sepsis entre otros, normalmente causados por patógenos oportunistas siendo uno de los microorganismos más comunes el Staphylococcus aures. En base a ello, se realizó un análisis retrospectivo de las historias clínicas de una cohorte HHT para comprobar la incidencia de determinadas infecciones, de cuyos resultados podemos concluir que en la población HHT española, se observa una mayor incidencia de determinados procesos infecciosos en comparación con la población general. Además de las infecciones, la expresión de endoglina y ALK1 durante la transición monocito-macrofágo hace sospechar que la respuesta inmunitaria puede estar afectada en el paciente HHT. Por ello, el siguiente paso fue analizar los niveles de expresión génica mediante microarrays de ADN en macrófagos de pacientes HHT diferenciados a partir de monocitos de sangre periférica. De ellos se concluye que 27 genes están inducidos mientras que 185 están reprimidos, respecto a controles no HHT. Dentro del listado de genes reprimidos, comprobamos que una gran parte interviene en procesos de la respuesta inmune. Estos estudios se complementaron con la evaluación de la producción de citoquinas por parte de la fracción MNC de pacientes HHT.Para profundizar en el estudio de la respuesta inmunitaria en el paciente de HHT, se generó un ratón deficiente en endoglina en macrófagos (Engfl/flLysMCre). Tras su caracterización, se realizaron ensayos para evaluar la respuesta inmunitaria. Los resultados de supervivencia a un shock séptico sugieren que la expresión de endoglina en el macrófago es importante para la iniciación de la respuesta inmune frente a infecciones. Además de esto, hemos observado que el reclutamiento de células inflamatorias tras inducir una peritonitis y la fagocitosis, son procesos que se ven afectados en ausencia de la expresión de endoglina en macrófagos. Tras la evaluación de algunos genes diana de TGF-β en los macrófagos peritoneales, postulamos que endoglina y ALK1 modulan el sistema de señalización de TGF-β en monocitos en diferenciación y macrófagos maduros. De este modo, la haploinsuficiencia en endoglina y ALK1 dará lugar a una disfunción de la respuesta inmune, que sería la causa subyacente a la mayor incidencia de enfermedades infecciosas reportadas en la literatura y también descritas en la presente tesis. En conjunto, los datos indican que sería necesario, a nivel clínico, ampliar la sintomatología clásica vascular asociada al síndrome de HHT,m añadiendo una inmunodeficiencia o fallo de la respuesta inmune innata. Incluyendo estos factores en el seguimiento de los pacientes HHT, se podría mejorar su asesoramiento y supervisión por parte del personal clínico y, prevenir la aparición de determinadas infecciones o bien en su defecto, apaliar las complicaciones serias de infecciones severas como los abscesos cerebrales y las bacteremias.Este trabajo ha sido realizado con cargo a los siguientes proyectos de investigación: SAF2005-01090 del Ministerio de Educación y Ciencia Periodo 2005-2008. SAF2008-01218 del Ministerio de Educación y Ciencia. Periodo 2009-2011. Contrato predoctoral (CIBERER) del Instituto de Salud Carlos III (ISCIII-CB06/07/0038Peer reviewe

Different Effects of Low Selenite and Selenium-Nanoparticle Supplementation on Adipose Tissue Function and Insulin Secretion in Adolescent Male Rats

Adolescence is a period of intense growth and endocrine changes, and obesity and insulinresistance processes during this period have lately been rising. Selenium (Se) homeostasis is related to lipid metabolism depending on the form and dose of Se. This study tests the actions of low-dose selenite and Se nanoparticles (SeNPs) on white (WAT) and brown adipose tissue (BAT) deposition, insulin secretion, and GPx1, IRS-1 and FOXO3a expression in the WAT of adolescent rats as regards oxidative stress, adipocyte length and adipokine secretion. Four groups of male adolescent rats were treated: control (C), low selenite supplementation (S), low SeNP supplementation (NS) and moderate SeNP supplementation (NSS). Supplementation was received orally through water intake; NS and NSS rats received two- and tenfold more Se than C animals, respectively. SeNPs were obtained by reducing Se tetrachloride in the presence of ascorbic acid. For the first time in vivo, it was demonstrated that low selenite supplementation contributed to increased adipogenesis via the insulin signaling pathway and LCN2 modulation, while low SeNP administration prevented fat depots inWAT via the decrease in insulin signaling and FOXO3a autophagy inWAT, lowering inflammation. These effects were independent of GPx1 expression or activity in WAT. These findings provide data for dietary approaches to prevent obesity and/or anorexia during adolescence. These findings may be relevant to future studies looking at a nutritional approach aimed at pre-venting obesity and/or anorexia in adolescence.Junta de AndaluciaFEDER projects funds US-1380878Spanish Government PID2019-109371GB-I00VII Plan Propio de Investigacion y Transferencia-University of Seville 2022 2022/00000332 2022/0000027

Inflammation and oxidative stress, the links between obesity and COVID-19: a narrative review

COVID-19, an acute respiratory disease caused by SARS-CoV-2, has rapidly become a pandemic. On the other hand, obesity is also reaching dramatic dimensions and it is a risk factor for morbidity and premature mortality. Obesity has been linked to a high risk of serious-associated complications to COVID-19, due to the increased risk of concomitant chronic diseases, which highlights the health public relevance of the topic. Obese subjects have a pro-inflammatory environment, which can further exacerbate COVID-19-induced inflammation and oxidative stress, explaining the increased risk of serious complications in these patients. Another factor that favors infection in obese patients is the high expression of ACE2 receptors in the adipose tissue. The negative impact of COVID-19 in obesity is also associated with a decrease in respiratory function, the concurrence of multiple comorbidities, a low-degree chronic inflammatory state, immunocompromised situation, and therefore a higher rate of hospitalization, mechanical ventilation, in-hospital complications such as pneumonia, and death. In this review, the link between obesity and COVID-19 was analyzed, exploring the potential common mechanisms in both diseases, with special attention to oxidative stress and inflammation, due to the crucial role of both pathways in the development of the disease

Reliability and Validity Study of the Chamorro Assisted Gait Scale for People with Sprained Ankles, Walking with Forearm Crutches

Objective The aim of this study was to design and validate a functional assessment scale for assisted gait with forearm crutches (Chamorro Assisted Gait Scale—CHAGS) and to assess its reliability in people with sprained ankles. Design Thirty subjects who suffered from sprained ankle (anterior talofibular ligament first and second degree) were included in the study. A modified Delphi technique was used to obtain the content validity. The selected items were: pelvic and scapular girdle dissociation(1), deviation of Center of Gravity(2), crutch inclination(3), steps rhythm(4), symmetry of step length(5), cross support(6), simultaneous support of foot and crutch(7), forearm off(8), facing forward(9) and fluency(10). Two raters twice visualized the gait of the sample subjects which were recorded. The criterion-related validity was determined by correlation between CHAGS and Coding of eight criteria of qualitative gait analysis (Viel Coding). Internal consistency and inter and intra-rater reliability were also tested. Results CHAGS obtained a high and negative correlation with Viel Coding. We obtained a good internal consistency and the intra-class correlation coefficients oscillated between 0.97 and 0.99, while the minimal detectable changes were acceptable. Conclusion CHAGS scale is a valid and reliable tool for assessing assisted gait with crutches in people with sprained ankles to perform partial relief of lower limbs.Telefonica Chair “Intelligence in Networks” of the University of Seville, Spai

Adipose Tissue Homeostasis Orchestrates the Oxidative, Energetic, Metabolic and Endocrine Disruption Induced by Binge Drinking in Adolescent Rats

Binge drinking (BD) is the most common alcohol consumption model for adolescents, and has recently been related to the generation of high oxidation and insulin resistance (IR). White adipose tissue (WAT) is a target organ for insulin action that regulates whole-body metabolism by secreting adipokines. The present study aimed to analyse the oxidative, inflammatory, energetic and endocrine profile in the WAT of BD-exposed adolescent rats, to obtain an integrative view of insulin secretion and WAT in IR progression. Two groups of male adolescent rats were used: control (n = 8) and BD (n = 8). An intermittent i.p. BD model (20% v/v) was used during 3 consecutive weeks. BD exposure led to a pancreatic oxidative imbalance, which was joint to high insulin secretion by augmenting deacetylase sirtuin-1 (SIRT-1) pancreatic expression and serum adipsin levels. However, BD rats had hyperglycaemia and high homeostasis model assessment of insulin resistance value (HOMA-IR). BD exposure in WAT increased lipid oxidation, as well as decreased insulin receptor substrate 1 (IRS-1) and AKT expression, sterol regulatory element-binding protein 1 (SREBP1), forkhead box O3A (FOXO3a) and peroxisome proliferator-activated receptor γ (PPARγ), and adipocyte size. BD also affected the expression of proteins related to energy balance, such as SIRT-1 and AMP activated protein kinase (AMPK), affecting the adipokine secretion profile (increasing resistin/adiponectin ratio). BD altered the entire serum lipid profile, increasing the concentration of free fatty acids. In conclusion, BD led to an oxidative imbalance and IR process in WAT, which modified the energy balance in this tissue, decreasing the WAT lipogenic/lipolytic ratio, affecting adipokine secretion and the systemic lipid profile, and contributing to the progression of IR. Therefore, WAT is key in the generation of metabolic and endocrine disruption after BD exposure during adolescence in rats. (Figure presented.). Key points: Adolescent rat binge drinking (BD) exposure leads to hepatic and systemic oxidative stress (OS) via reactive oxygen species generation, causing hepatic insulin resistance (IR) and altered energy metabolism. In the present study, BD exposure in adolescent rats induces OS in the pancreas, with increased insulin secretion despite hyperglycaemia, indicating a role for IR in white adipose tissue (WAT) homeostasis. In WAT, BD produces IR and an oxidative and energetic imbalance, triggering an intense lipolysis where the serum lipid profile is altered and free fatty acids are increased, consistent with liver lipid accumulation and steatosis. BD exposure heightens inflammation in WAT, elevating pro-inflammatory and reducing anti-inflammatory adipokines, favouring cardiovascular damage. This research provides a comprehensive view of how adolescent BD in rats impacts liver, WAT and pancreas homeostasis, posing a risk for future cardiometabolic complications in adulthood.Junta de Andalucía CTS-193, USE-22 212-

Mice lacking endoglin in macrophages show an impaired immune response

24 p.-9 fig.-1 tab. Ojeda Fernández, Luisa et al.Endoglin is an auxiliary receptor for members of the TGF-β superfamily and plays an important role in the homeostasis of the vessel wall. Mutations in endoglin gene (ENG) or in the closely related TGF-β receptor type I ACVRL1/ALK1 are responsible for a rare dominant vascular dysplasia, the Hereditary Hemorrhagic Telangiectasia (HHT), or Rendu-OslerWeber syndrome. Endoglin is also expressed in human macrophages, but its role in macrophage function remains unknown. In this work, we show that endoglin expression is triggered during the monocyte-macrophage differentiation process, both in vitro and during the in vivo differentiation of blood monocytes recruited to foci of inflammation in wild-type C57BL/6 mice. To analyze the role of endoglin in macrophages in vivo, an endoglin myeloid lineage specific knock-out mouse line (Engfl/flLysMCre) was generated. These mice show a predisposition to develop spontaneous infections by opportunistic bacteria. Engfl/flLysMCre mice also display increased survival following LPS-induced peritonitis, suggesting a delayed immune response. Phagocytic activity is impaired in peritoneal macrophages, altering one of the main functions of macrophages which contributes to the initiation of the immune response. We also observed altered expression of TGF-β1 target genes in endoglin deficient peritoneal macrophages. Overall, the altered immune activity of endoglin deficient macrophages could help to explain the higher rate of infectious diseases seen in HHT1 patients.This work was funded by: Ministerio de Economía y Competitividad of Spain (SAF2011-23475 to LMB; SAF2013-43421-R and SAF2010- 19222 to CB.Peer reviewe

Effectiveness of a strategy that uses educational games to implement clinical practice guidelines among Spanish residents of family and community medicine (e-EDUCAGUIA project):A clinical trial by clusters

This study was funded by the Fondo de Investigaciones Sanitarias FIS Grant Number PI11/0477 ISCIII.-REDISSEC Proyecto RD12/0001/0012 AND FEDER Funding.Background: Clinical practice guidelines (CPGs) have been developed with the aim of helping health professionals, patients, and caregivers make decisions about their health care, using the best available evidence. In many cases, incorporation of these recommendations into clinical practice also implies a need for changes in routine clinical practice. Using educational games as a strategy for implementing recommendations among health professionals has been demonstrated to be effective in some studies; however, evidence is still scarce. The primary objective of this study is to assess the effectiveness of a teaching strategy for the implementation of CPGs using educational games (e-learning EDUCAGUIA) to improve knowledge and skills related to clinical decision-making by residents in family medicine. The primary objective will be evaluated at 1 and 6months after the intervention. The secondary objectives are to identify barriers and facilitators for the use of guidelines by residents of family medicine and to describe the educational strategies used by Spanish teaching units of family and community medicine to encourage implementation of CPGs. Methods/design: We propose a multicenter clinical trial with randomized allocation by clusters of family and community medicine teaching units in Spain. The sample size will be 394 residents (197 in each group), with the teaching units as the randomization unit and the residents comprising the analysis unit. For the intervention, both groups will receive an initial 1-h session on clinical practice guideline use and the usual dissemination strategy by e-mail. The intervention group (e-learning EDUCAGUIA) strategy will consist of educational games with hypothetical clinical scenarios in a virtual environment. The primary outcome will be the score obtained by the residents on evaluation questionnaires for each clinical practice guideline. Other included variables will be the sociodemographic and training variables of the residents and the teaching unit characteristics. The statistical analysis will consist of a descriptive analysis of variables and a baseline comparison of both groups. For the primary outcome analysis, an average score comparison of hypothetical scenario questionnaires between the EDUCAGUIA intervention group and the control group will be performed at 1 and 6months post-intervention, using 95% confidence intervals. A linear multilevel regression will be used to adjust the model. Discussion: The identification of effective teaching strategies will facilitate the incorporation of available knowledge into clinical practice that could eventually improve patient outcomes. The inclusion of information technologies as teaching tools permits greater learning autonomy and allows deeper instructor participation in the monitoring and supervision of residents. The long-term impact of this strategy is unknown; however, because it is aimed at professionals undergoing training and it addresses prevalent health problems, a small effect can be of great relevance. Trial registration: ClinicalTrials.gov: NCT02210442.Publisher PDFPeer reviewe

Efficacy of clozapine versus standard treatment in adult individuals with intellectual disability and treatment-resistant psychosis (CLOZAID): study protocol of a multicenter randomized clinical trial

BackgroundIntellectual disability (ID) affects approximately 1% of the worldwide population and individuals with ID have a higher comorbidity with mental illness, and specifically psychotic disorders. Unfortunately, among individuals with ID, limited research has been conducted since ID individuals are usually excluded from mental illness epidemiological studies and clinical trials. Here we perform a clinical trial to investigate the effectiveness of clozapine in the treatment of resistant psychosis in individuals with ID. The article highlights the complexity of diagnosing and treating psychopathological alterations associated with ID and advocates for more rigorous research in this field.MethodsA Phase IIB, open-label, randomized, multicenter clinical trial (NCT04529226) is currently ongoing to assess the efficacy of oral clozapine in individuals diagnosed with ID and suffering from treatment-resistant psychosis. We aim to recruit one-hundred and fourteen individuals (N=114) with ID and resistant psychosis, who will be randomized to TAU (treatment as usual) and treatment-with-clozapine conditions. As secondary outcomes, changes in other clinical scales (PANSS and SANS) and the improvement in functionality, assessed through changes in the Euro-QoL-5D-5L were assessed. The main outcome variables will be analyzed using generalized linear mixed models (GLMM), assessing the effects of status variable (TAU vs. Clozapine), time, and the interaction between them.DiscussionThe treatment of resistant psychosis among ID individuals must be directed by empirically supported research. CLOZAID clinical trial may provide relevant information about clinical guidelines to optimally treat adults with ID and treatment-resistant psychosis and the benefits and risks of an early use of clozapine in this underrepresented population in clinical trials.Trial registrationClinicaltrials.gov: NCT04529226. EudraCT: 2020-000091-37