The protein quality control system in motoneuron diseases

Spinal and bulbar muscular atrophy (SBMA) is a motoneuronal diseases caused by an elogated polyglutamine (polyQ) tract in the androgen receptor (AR). The polyQ expansion causes the AR protein to misfold and the binding with the ligand testosterone triggers a cascade of events, including ARpolyQ aggregation, that led to motoneuron death. The intracellular accumulation of misfolded ARpolyQ both altered the protein quality control system (PQC) and impaired the protective mechanisms deputed to refolding and clearance of misfolded proteins. In PQC, the molecular chaperones allow the refolding or the clearance of the misfolded proteins through the Ubiquitin Proteasome system (UPS) or the autophagic pathway. Moreover, emerging evidence reveal that ARpolyQ toxicity is not related only to motoneuron degeneration but also skeletal muscle damage plays a primary role in SBMA. AIM: The aim of the study was both to unravell the contribution of PQC in SBMA and to find molecular and pharmacological approaches for modulating PQC as potential therapeutic target. Methods: Western blot and filter retardation assay were used to analyse the biochemical properties of ARpolyQ and the protein level of PQC markers. RT-qPCR was used to quantify the mRNA expression of PQC genes in presence of ARpolyQ. Results: In SBMA motoneuronal cell line, we demonstrated that both UPS and autophagic pathway are impaired or blocked, leading to ARpolyQ accumulation into the aggregates. Moreover, analysis in SBMA animal model showed that in the spinal cord and in the skeletal muscle, the PQC could differ considerably in how degrading the mutant and misfolded ARpolyQ. In these conditions of PQC impairment we tested, in SBMA cell model, the overexpression of the small heat shock protein B8 (HspB8), involved in the autophagic pathway. HpB8 led to the autophagic removal of misfolded ARpolyQ, restorating the intracellular autophagic flux. Interestingly, we found that trehalose, a known autophagic stimulator, was able to induce the HspB8 expression and to facilitate the ARpolyQ clearance. Then, we tested the combined treatment of trehalose with Bicalutamide, an antiandrogen. Bicalutamide is able to slow down AR nuclear translocation and to retain it into the cytoplasm, where the autophagic pathway is active. Bicalutamide and trehalose showed synergic activity in the degradation of ARpolyQ. Conclusions: the PQC plays a crucial role in SBMA, the modulation of its activity with trehalose and Bicalutamide might be a promising approach for this no cure disease

New approach methodologies to enhance human health risk assessment of immunotoxic properties of chemicals: a PARC (Partnership for the Assessment of Risk from Chemicals) project

As a complex system governing and interconnecting numerous functions within the human body, the immune system is unsurprisingly susceptible to the impact of toxic chemicals. Toxicants can influence the immune system through a multitude of mechanisms, resulting in immunosuppression, hypersensitivity, increased risk of autoimmune diseases and cancer development. At present, the regulatory assessment of the immunotoxicity of chemicals relies heavily on rodent models and a limited number of Organisation for Economic Co-operation and Development (OECD) test guidelines, which only capture a fraction of potential toxic properties. Due to this limitation, various authorities, including the World Health Organization and the European Food Safety Authority have highlighted the need for the development of novel approaches without the use of animals for immunotoxicity testing of chemicals. In this paper, we present a concise overview of ongoing efforts dedicated to developing and standardizing methodologies for a comprehensive characterization of the immunotoxic effects of chemicals, which are performed under the EU-funded Partnership for the Assessment of Risk from Chemicals (PARC)

New approach methodologies to enhance human health risk assessment of immunotoxic properties of chemicals — a PARC (Partnership for the Assessment of Risk from Chemicals) project

As a complex system governing and interconnecting numerous functions within the human body, the immune system is unsurprisingly susceptible to the impact of toxic chemicals. Toxicants can influence the immune system through a multitude of mechanisms, resulting in immunosuppression, hypersensitivity, increased risk of autoimmune diseases and cancer development. At present, the regulatory assessment of the immunotoxicity of chemicals relies heavily on rodent models and a limited number of Organisation for Economic Co-operation and Development (OECD) test guidelines, which only capture a fraction of potential toxic properties. Due to this limitation, various authorities, including the World Health Organization and the European Food Safety Authority have highlighted the need for the development of novel approaches without the use of animals for immunotoxicity testing of chemicals. In this paper, we present a concise overview of ongoing efforts dedicated to developing and standardizing methodologies for a comprehensive characterization of the immunotoxic effects of chemicals, which are performed under the EU-funded Partnership for the Assessment of Risk from Chemicals (PARC)

Effect of angiotensin-converting enzyme inhibitor and angiotensin receptor blocker initiation on organ support-free days in patients hospitalized with COVID-19

IMPORTANCE Overactivation of the renin-angiotensin system (RAS) may contribute to poor clinical outcomes in patients with COVID-19. Objective To determine whether angiotensin-converting enzyme (ACE) inhibitor or angiotensin receptor blocker (ARB) initiation improves outcomes in patients hospitalized for COVID-19. DESIGN, SETTING, AND PARTICIPANTS In an ongoing, adaptive platform randomized clinical trial, 721 critically ill and 58 non–critically ill hospitalized adults were randomized to receive an RAS inhibitor or control between March 16, 2021, and February 25, 2022, at 69 sites in 7 countries (final follow-up on June 1, 2022). INTERVENTIONS Patients were randomized to receive open-label initiation of an ACE inhibitor (n = 257), ARB (n = 248), ARB in combination with DMX-200 (a chemokine receptor-2 inhibitor; n = 10), or no RAS inhibitor (control; n = 264) for up to 10 days. MAIN OUTCOMES AND MEASURES The primary outcome was organ support–free days, a composite of hospital survival and days alive without cardiovascular or respiratory organ support through 21 days. The primary analysis was a bayesian cumulative logistic model. Odds ratios (ORs) greater than 1 represent improved outcomes. RESULTS On February 25, 2022, enrollment was discontinued due to safety concerns. Among 679 critically ill patients with available primary outcome data, the median age was 56 years and 239 participants (35.2%) were women. Median (IQR) organ support–free days among critically ill patients was 10 (–1 to 16) in the ACE inhibitor group (n = 231), 8 (–1 to 17) in the ARB group (n = 217), and 12 (0 to 17) in the control group (n = 231) (median adjusted odds ratios of 0.77 [95% bayesian credible interval, 0.58-1.06] for improvement for ACE inhibitor and 0.76 [95% credible interval, 0.56-1.05] for ARB compared with control). The posterior probabilities that ACE inhibitors and ARBs worsened organ support–free days compared with control were 94.9% and 95.4%, respectively. Hospital survival occurred in 166 of 231 critically ill participants (71.9%) in the ACE inhibitor group, 152 of 217 (70.0%) in the ARB group, and 182 of 231 (78.8%) in the control group (posterior probabilities that ACE inhibitor and ARB worsened hospital survival compared with control were 95.3% and 98.1%, respectively). CONCLUSIONS AND RELEVANCE In this trial, among critically ill adults with COVID-19, initiation of an ACE inhibitor or ARB did not improve, and likely worsened, clinical outcomes. TRIAL REGISTRATION ClinicalTrials.gov Identifier: NCT0273570

Analytical Performance of Different Laboratory Methods for Measuring Susoctocog-Alfa

Recombinant porcine factor VIII (rpFVIII) is indicated for treating bleeding episodes in acquired haemophilia A, but there are few data regarding laboratory methods to adequately monitor treatment. This study involving three Italian laboratories aimed to evaluate the analytical performance of different assays for measuring rpFVIII. Five spiked rpFVIII samples (0.5–1.5 IU/mL) were analysed on three days, in triplicate, with eleven combinations of reagents (Werfen, Boston, MA, USA: SynthasIL and SynthaFax for one-stage assay, Chromogenix Coamatic FVIII for chromogenic assay), FVIII depleted plasmas (with or without von Willebrand factor—VWF) and calibrators (HemosIL human calibrator plasma, porcine calibrator diluted in FVIII deficient plasma with or without VWF). The assays were performed on ACL TOP analysers (Werfen, Boston, MA, USA). Intra- and inter-assay and inter-laboratory Coefficient of Variation (CV%) were calculated together with percentage of recovery (% recovery) on the expected value. The results showed that the reagent combinations reaching satisfactory analytical performance are: SynthasIL/human calibrator/deficient plasma+VWF (total recovery 99.4%, inter-laboratory CV 4.04%), SynthasIL/porcine calibrator/deficient plasma+VWF (total recovery 111%, inter-laboratory CV 2.75%) and Chromogenic/ porcine calibrator/deficient plasma+VWF (total recovery 96.6%, inter-laboratory CV 8.32%). This study highlights that the use of porcine standard (when available) and FVIII deficient plasma with VWF should be recommended

TX - Taller de Tesis - AR304 - 202101

Descripción: En este curso se desarrollará, mediante una propuesta arquitectónica, un proyecto escogido por el estudiante, en el que pueda acreditar que está en capacidad de hacer frente a los principales aspectos comprendidos en el desarrollo de un proyecto arquitectónico. En la primera etapa se establecen los criterios básicos de diseño de un pre-anteproyecto arquitectónico sustentado a partir de las condiciones: conceptuales, programáticas y de usuario, aspectos medio ambientales, urbano y paisajistas. En la segunda etapa se desarrolla un anteproyecto arquitectónico incluyendo además los criterios de las especialidades de estructuras, instalaciones eléctricas y sanitarias, y sistemas de evacuación. Propósito: El curso tiene como propósito la aprobación del anteproyecto con el cual el estudiante puede iniciar su Proyecto de Titulación Profesional. Busca contribuir al desarrollo de las competencias generales UPC: Comunicación Escrita, Comunicación Oral, Pensamiento Crítico, Razonamiento Cuantitativo, Manejo de la Información, Ciudadanía, Pensamiento Innovador y de las competencias específicas de la carrera: Diseño Fundamentado (que corresponde a los criterios NAAB PC2, PC3,PC5, PC8, SC5), Cultura Arquitectónica (que corresponde a los criterios NAAB1 PC4), Técnica y Construcción (que corresponde a los criterios NAAB1 SC1, SC4,SC6) y Gestión Profesional (que corresponde a los criterios NAAB1 PC6, SC2), todas en el nivel 3. Tiene como requisitos AR248 Gestión Inmobiliaria y HU61 inglés 5 y AR271 Lineamientos para el Proyecto Profesional y AR272 Seminario de Urbanismo y AR250 TIX - Taller de Ejercicio Profesional y aprobación por el director de la Carrera

TX - Taller de Tesis - AR304 - 202102

Descripción: En este curso se desarrollará, mediante una propuesta arquitectónica, un proyecto escogido por el estudiante, en el que pueda acreditar que está en capacidad de hacer frente a los principales aspectos comprendidos en el desarrollo de un proyecto arquitectónico. En la primera etapa se establecen los criterios básicos de diseño de un pre-anteproyecto arquitectónico sustentado a partir de las condiciones: conceptuales, programáticas y de usuario, aspectos medio ambientales, urbano y paisajistas. En la segunda etapa se desarrolla un anteproyecto arquitectónico incluyendo además los criterios de las especialidades de estructuras, instalaciones eléctricas y sanitarias, y sistemas de evacuación. Propósito: El curso tiene como propósito la aprobación del anteproyecto con el cual el estudiante puede iniciar su Proyecto de Titulación Profesional. Busca contribuir al desarrollo de las competencias generales UPC: Comunicación Escrita, Comunicación Oral, 3Pensamiento Crítico, Razonamiento Cuantitativo, Manejo de la Información, Ciudadanía, Pensamiento Innovador y de las competencias específicas de la carrera: Diseño Fundamentado (que corresponde a los criterios NAAB PC2, PC3,PC5, PC8, SC5), Cultura Arquitectónica (que corresponde a los criterios NAAB1 PC4), Técnica y Construcción (que corresponde a los criterios NAAB1 SC1, SC4,SC6) y Gestión Profesional (que corresponde a los criterios NAAB1 PC6, SC2), todas en el nivel 3. Tiene como requisitos AR248 Gestión Inmobiliaria y HU61 inglés 5 y AR271 Lineamientos para el Proyecto Profesional y AR272 Seminario de Urbanismo y AR250 TIX - Taller de Ejercicio Profesional y aprobación por el director de la Carrera

Prosafe: a european endeavor to improve quality of critical care medicine in seven countries

BACKGROUND: long-lasting shared research databases are an important source of epidemiological information and can promote comparison between different healthcare services. Here we present ProsaFe, an advanced international research network in intensive care medicine, with the focus on assessing and improving the quality of care. the project involved 343 icUs in seven countries. all patients admitted to the icU were eligible for data collection. MetHoDs: the ProsaFe network collected data using the same electronic case report form translated into the corresponding languages. a complex, multidimensional validation system was implemented to ensure maximum data quality. individual and aggregate reports by country, region, and icU type were prepared annually. a web-based data-sharing system allowed participants to autonomously perform different analyses on both own data and the entire database. RESULTS: The final analysis was restricted to 262 general ICUs and 432,223 adult patients, mostly admitted to Italian units, where a research network had been active since 1991. organization of critical care medicine in the seven countries was relatively similar, in terms of staffing, case mix and procedures, suggesting a common understanding of the role of critical care medicine. conversely, icU equipment differed, and patient outcomes showed wide variations among countries. coNclUsioNs: ProsaFe is a permanent, stable, open access, multilingual database for clinical benchmarking, icU self-evaluation and research within and across countries, which offers a unique opportunity to improve the quality of critical care. its entry into routine clinical practice on a voluntary basis is testimony to the success and viability of the endeavor