Allergic inflammation triggers dyslipidemia via IgG signalling.

BACKGROUND: Allergic diseases begin early in life and are often chronic, thus creating an inflammatory environment that may precede or exacerbate other pathologies. In this regard, allergy has been associated to metabolic disorders and with a higher risk of cardiovascular disease, but the underlying mechanisms remain incompletely understood. METHODS: We used a murine model of allergy and atherosclerosis, different diets and sensitization methods, and cell-depleting strategies to ascertain the contribution of acute and late phase inflammation to dyslipidemia. Untargeted lipidomic analyses were applied to define the lipid fingerprint of allergic inflammation at different phases of allergic pathology. Expression of genes related to lipid metabolism was assessed in liver and adipose tissue at different times post-allergen challenge. Also, changes in serum triglycerides (TGs) were evaluated in a group of 59 patients ≥14 days after the onset of an allergic reaction. RESULTS: We found that allergic inflammation induces a unique lipid signature that is characterized by increased serum TGs and changes in the expression of genes related to lipid metabolism in liver and adipose tissue. Alterations in blood TGs following an allergic reaction are independent of T-cell-driven late phase inflammation. On the contrary, the IgG-mediated alternative pathway of anaphylaxis is sufficient to induce a TG increase and a unique lipid profile. Lastly, we demonstrated an increase in serum TGs in 59 patients after undergoing an allergic reaction. CONCLUSION: Overall, this study reveals that IgG-mediated allergic inflammation regulates lipid metabolism.

Protein homeostasis maintained by HOOK1 levels promotes the tumorigenic and stemness properties of ovarian cancer cells through reticulum stress and autophagy.

BACKGROUND: Ovarian cancer has a high mortality rate mainly due to its resistance to currently used therapies. This resistance has been associated with the presence of cancer stem cells (CSCs), interactions with the microenvironment, and intratumoral heterogeneity. Therefore, the search for new therapeutic targets, particularly those targeting CSCs, is important for improving patient prognosis. HOOK1 has been found to be transcriptionally altered in a substantial percentage of ovarian tumors, but its role in tumor initiation and development is still not fully understood. METHODS: The downregulation of HOOK1 was performed in ovarian cancer cell lines using CRISPR/Cas9 technology, followed by growth in vitro and in vivo assays. Subsequently, migration (Boyden chamber), cell death (Western-Blot and flow cytometry) and stemness properties (clonal heterogeneity analysis, tumorspheres assay and flow cytometry) of the downregulated cell lines were analysed. To gain insights into the specific mechanisms of action of HOOK1 in ovarian cancer, a proteomic analysis was performed, followed by Western-blot and cytotoxicity assays to confirm the results found within the mass spectrometry. Immunofluorescence staining, Western-blotting and flow cytometry were also employed to finish uncovering the role of HOOK1 in ovarian cancer. RESULTS: In this study, we observed that reducing the levels of HOOK1 in ovarian cancer cells reduced in vitro growth and migration and prevented tumor formation in vivo. Furthermore, HOOK1 reduction led to a decrease in stem-like capabilities in these cells, which, however, did not seem related to the expression of genes traditionally associated with this phenotype. A proteome study, along with other analysis, showed that the downregulation of HOOK1 also induced an increase in endoplasmic reticulum stress levels in these cells. Finally, the decrease in stem-like properties observed in cells with downregulated HOOK1 could be explained by an increase in cell death in the CSC population within the culture due to endoplasmic reticulum stress by the unfolded protein response. CONCLUSION: HOOK1 contributes to maintaining the tumorigenic and stemness properties of ovarian cancer cells by preserving protein homeostasis and could be considered an alternative therapeutic target, especially in combination with inducers of endoplasmic reticulum or proteotoxic stress such as proteasome inhibitors.

A comparison of four self-controlled study designs in an analysis of COVID-19 vaccines and myocarditis using five European databases.

INTRODUCTION: The aim of this study was to assess the possible extent of bias due to violation of a core assumption (event-dependent exposures) when using self-controlled designs to analyse the association between COVID-19 vaccines and myocarditis. METHODS: We used data from five European databases (Spain: BIFAP, FISABIO VID, and SIDIAP; Italy: ARS-Tuscany; England: CPRD Aurum) converted to the ConcePTION Common Data Model. Individuals who experienced both myocarditis and were vaccinated against COVID-19 between 1 September 2020 and the end of data availability in each country were included. We compared a self-controlled risk interval study (SCRI) using a pre-vaccination control window, an SCRI using a post-vaccination control window, a standard SCCS and an extension of the SCCS designed to handle violations of the assumption of event-dependent exposures. RESULTS: We included 1,757 cases of myocarditis. For analyses of the first dose of the Pfizer vaccine, to which all databases contributed information, we found results consistent with a null effect in both of the SCRI and extended SCCS, but some indication of a harmful effect in a standard SCCS. For the second dose, we found evidence of a harmful association for all study designs, with relatively similar effect sizes (SCRI pre = 1.99, 1.40 - 2.82; SCRI post 2.13, 95 %CI - 1.43, 3.18; standard SCCS 1.79, 95 %CI 1.31 - 2.44, extended SCCS 1.52, 95 %CI = 1.08 - 2.15). Adjustment for calendar time did not change these conclusions. Findings using all designs were also consistent with a harmful effect following a second dose of the Moderna vaccine. CONCLUSIONS: In the context of the known association between COVID-19 vaccines and myocarditis, we have demonstrated that two forms of SCRI and two forms of SCCS led to largely comparable results, possibly because of limited violation of the assumption of event-dependent exposures.

Risk of herpes zoster in adults with SARS-CoV-2 infection in Spain: A population-based, retrospective cohort study.

OBJECTIVES: We aimed to compare the risk of herpes zoster (HZ) in adults with and without laboratory-confirmed severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) infection. METHODS: This retrospective dynamic cohort study analyzed data from a public healthcare database in Spain between November 2020 and October 2021. The main outcome was incident cases of HZ in individuals ≥18-year-old. Relative risk (RR) of HZ in SARS-CoV-2-confirmed versus SARS-CoV-2-free individuals was estimated by a multivariable negative binomial regression adjusted by age, sex, and comorbidities. RESULTS: Data from 4,085,590 adults were analyzed. The overall HZ incidence rate in adults was 5.76 (95% confidence interval [CI], 5.66-5.85) cases per 1000 person-years. Individuals ≥18-year-old with SARS-CoV-2-confirmed infection had a 19% higher risk of developing HZ versus SARS-CoV-2-free ≥18-year-olds (adjusted RR = 1.19; 95% CI, 1.09-1.29); this percentage was 16% (adjusted RR = 1.16; 95% CI, 1.05-1.29) in ≥50-year-olds. Severe (hospitalized) cases of SARS-CoV-2 infection had a 64% (if ≥18 years old) or 44% (if ≥50 years old) higher risk of HZ versus nonhospitalized cases. CONCLUSION: These results support an association between SARS-CoV-2 infection and HZ, with a greater HZ risk in severe cases of SARS-CoV-2 infection.

Proyecto ECHO Uruguay: fortalecimiento de la formación de profesionales en el abordaje del cáncer ginecológico

En Uruguay existe una desigual distribución de médicos entre la capital y el resto del país, determinando que los pacientes deban ser referidos a centros asistenciales fuera del área en la que residen. El Proyecto ECHO (del inglés: Extension for Community Healthcare Outcomes) busca mejorar el acceso a atención especializada para poblaciones rurales mediante la utilización de tecnologías de la comunicación, democratizando el conocimiento. Objetivo: evaluar los resultados en lo referente a competencias y habilidades profesionales en los participantes de las teleclínicas ECHO sobre cáncer ginecológico en Uruguay. Método: evaluación retrospectiva de impacto con línea de base y línea de impacto mediante un censo vía web. Período: setiembre 2020- mayo/2021. Se relevaron 22 variables, 14 indicadores dependientes que miden autopercepciones sobre incremento de capacidades en forma retrospectiva a partir de una escala Likert de 5 valores y, 8 independientes que abordan aspectos generales de la población. Teniendo en cuenta que no hay una distribución normal se aplica el test no paramétrico de Wilkoxon. Resultados: se obtuvieron 36 respuestas. Los resultados, muestran que todos los pares tienen una significación bilateral, pudiendo afirmar que existe una diferencia significativa entre las capacidades previas y posteriores a la participación en el programa ECHO. Conclusiones: se objetiva una mejora en la autopercepción de las capacidades de los participantes luego de la implementación de las teleclínicas ECHO.

Summary: In Uruguay there is an unequal distribution of doctors between the capital and the rest of the country, determining that patients must be referred to healthcare centers outside the area in which they reside. The ECHO (Extension for Community Healthcare Outcomes) Project seeks to improve access to specialized care for rural populations through the use of communication technologies, democratizing knowledge. Objective: to evaluate the results regarding professional competencies and skills in the participants of the ECHO teleclinics on gynecological cancer in Uruguay. Method: retrospective impact evaluation with baseline and impact line through a web census. Period: September 2020- May/2021. 22 variables were surveyed, 14 dependent indicators that measure self-perceptions of increased capabilities retrospectively based on a Likert scale of 5 values, and 8 independent indicators that address general aspects of the population. Taking into account that there is no normal distribution, the non-parametric Wilkoxon test is applied. Results: 36 responses were obtained. The results show that all pairs have a bilateral significance, being able to affirm that there is a significant difference between the capabilities before and after participation in the ECHO program. Conclusions: an improvement in the self-perception of the participants' capabilities was observed after the implementation of the ECHO teleclinics.

No Uruguai existe uma distribuição desigual de médicos entre a capital e o resto do país, determinando que os pacientes sejam encaminhados para centros de saúde fora da área em que residem. O Projeto ECHO (de Inglês: Extension for Community Healthcare Outcomes)procura melhorar o acesso a cuidados especializados para as populações rurais através do uso de tecnologias de comunicação, democratizando o conhecimento. Objetivo: avaliar os resultados relativos às competências e habilidades profissionais dos participantes das teleclínicas ECHO sobre câncer ginecológico no Uruguai. Método: avaliação retrospectiva de impacto com linha de base e linha de impacto através de censo web. Período: Setembro 2020- Maio/2021. Foram levantadas 22 variáveis, 14 indicadores dependentes que medem retrospectivamente as autopercepções de aumento de capacidades com base em uma escala Likert de 5 valores, e 8 indicadores independentes que abordam aspectos gerais da população. Levando em consideração que não existe distribuição normal, aplica-se o teste não paramétrico de Wilkoxon. Resultados: foram obtidas 36 respostas. Os resultados mostram que todos os pares têm um significado bilateral, podendo afirmar que existe uma diferença significativa entre as capacidades antes e depois da participação no programa ECHO. Conclusões: observou-se melhora na autopercepção das capacidades dos participantes após a implantação das teleclínicas ECHO.

ZMYM2 controls human transposable element transcription through distinct co-regulatory complexes.

ZMYM2 is a zinc finger transcriptional regulator that plays a key role in promoting and maintaining cell identity. It has been implicated in several diseases such as congenital anomalies of the kidney where its activity is diminished and cancer where it participates in oncogenic fusion protein events. ZMYM2 is thought to function through promoting transcriptional repression and here we provide more evidence to support this designation. Here we studied ZMYM2 function in human cells and demonstrate that ZMYM2 is part of distinct chromatin-bound complexes including the established LSD1-CoREST-HDAC1 corepressor complex. We also identify new functional and physical interactions with ADNP and TRIM28/KAP1. The ZMYM2-TRIM28 complex forms in a SUMO-dependent manner and is associated with repressive chromatin. ZMYM2 and TRIM28 show strong functional similarity and co-regulate a large number of genes. However, there are no strong links between ZMYM2-TRIM28 binding events and nearby individual gene regulation. Instead, ZMYM2-TRIM28 appears to regulate genes in a more regionally defined manner within TADs where it can directly regulate co-associated retrotransposon expression. We find that different types of ZMYM2 binding complex associate with and regulate distinct subclasses of retrotransposons, with ZMYM2-ADNP complexes at SINEs and ZMYM2-TRIM28 complexes at LTR elements. We propose a model whereby ZMYM2 acts directly through retrotransposon regulation, which may then potentially affect the local chromatin environment and associated coding gene expression.

Gut Microbiota Associated with Clostridioides difficile Carriage in Three Clinical Groups (Inflammatory Bowel Disease, C. difficile Infection and Healthcare Workers) in Hospital Field.

Clostridioides difficile is an anaerobic spore-forming Gram-positive bacterium. C. difficile carriage and 16S rDNA profiling were studied in three clinical groups at three different sampling times: inflammatory bowel disease (IBD) patients, C. difficile infection (CDI) patients and healthcare workers (HCWs). Diversity analysis was realized in the three clinical groups, the positive and negative C. difficile carriage groups and the three analysis periods. Concerning the three clinical groups, ß-diversity tests showed significant differences between them, especially between the HCW group and IBD group and between IBD patients and CDI patients. The Simpson index (evenness) showed a significant difference between two clinical groups (HCWs and IBD). Several genera were significantly different in the IBD patient group (Sutterella, Agathobacter) and in the CDI patient group (Enterococcus, Clostridioides). Concerning the positive and negative C. difficile carriage groups, ß-diversity tests showed significant differences. Shannon, Simpson and InvSimpson indexes showed significant differences between the two groups. Several genera had significantly different relative prevalences in the negative group (Agathobacter, Sutterella, Anaerostipes, Oscillospira) and the positive group (Enterococcus, Enterobacteriaceae_ge and Enterobacterales_ge). A microbiota footprint was detected in C. difficile-positive carriers. More experiments are needed to test this microbiota footprint to see its impact on C. difficile infection.

Caveolin Delivered by Ultrasound-Mediated Microbubble Destruction Prevents Endothelial Cell Proliferation.

Introduction: The nitric oxide synthase (eNOS) is an important regulator of vascular homeostasis. eNOS is modulated by intracellular mechanisms that include protein-protein interaction with Caveolin-1 (Cav). Cav binds to and impairs eNOS activation reducing vascular permeability and angiogenesis. Blocking of eNOS by Cav has been proposed as therapeutic antiangiogenic approach. However, the efficient and controlled delivery of the peptide requires to be solved. Methods: The effect of antennapedia (AP)-Cav loaded into microbubbles (MBs) and delivered by ultrasound-mediated microbubble destruction (UMMD) into brain endothelial cells (bEnd.3 cells) was evaluated on NO production using DAF2-DA, cell migration assessed by the wound healing assay, cell proliferation with BrdU, and ex-vivo angiogenesis in rat aortic rings. Results: An enhanced inhibitory effect of AP-Cav was observed on cells treated with UMMD. MBs and ultrasound disruption delivery of AP-Cav increased acetylcholine-induced NO release, wound healing, cell proliferation, and angiogenesis inhibition on bEnd.3 cells, compared to free AP-Cav administration. Conclusion: We demonstrated that the delivery of Cav via AP-Cav-loaded MBs and UMMD may be an administration method for Cav that would increase its therapeutic potential by enhancing efficacy and cellular specificity.

Spatio-temporal assessment of soil properties immediately and eight months after a high intensity-controlled burn in the south of Spain.

In recent years, the use of fire as a means by which to manage forest ecosystems has become more frequent in Europe. Fire has a significant impact on the soil, and it is therefore necessary to understand how controlled burns affect this invaluable resource. The purpose of this study was to evaluate the main alterations in the physical-chemical and biological properties of the soil because of a high intensity-controlled burn in "Los Boquerones" area (Villaviciosa de Córdoba, Spain). Additionally, we assessed the spatial heterogeneity of the alterations of different soil properties. A grid of 12 points was established on a hillside in Sierra Morena (Córdoba). Thermocouples were placed at each point, and soil samples were collected at two depths (0-2 cm and 2-5 cm) before burning, immediately after burning and eight months later. Soil pH, electrical conductivity, nutrient content and/or availability, among others, and their spatio-temporal variations were analysed. Soil pH, increased in the first centimetres of the soil (0-2 cm) immediately after burning up to >2 units, and the increase was maintained eight months following the burn. Additionally, the high-intensity burn had a positive short-term effect on some of the soil properties, such as nutrient availability for plants, which was considerably increased. The magnitude of the alterations in the soil indicators assessed was spatially explained by the behaviour of the fire during the controlled burning. The burn also had both direct and indirect effects on soil microorganisms. In conclusion, the possible immediate and short-term effects of burning on the soil resource should be considered for a more holistic management of fire in forest ecosystems, as its functionality and capacity to provide ecosystem services is largely altered by these events as a function of their intensity.

Cuestionarios de adherencia al tratamiento antidiabético oral en pacientes con Diabetes Mellitus 2: revisión sistemática / Adherence questionnaires to oral antidiabetic treatment in patients with Diabetes Mellitus 2: systematic review

Introducción: La adherencia a los antidiabéticos orales (ADOs) en pacientes con Diabetes Mellitus tipo 2 (DM2) puede afectar al control de la enfermedad y por tanto, es necesario evaluarla. Para ello se necesitan herramientas rápidas, sencillas, válidas y fiables. De manera que el objetivo de esta revisión fue identificar aquellos cuestionarios validados que se usan para medir la adherencia a los ADOs en la práctica clínica. Método: se llevó a cabo una búsqueda bibliográfica en las bases de datos Medline, Scopus y LILACS. La selección de artículos la realizaron dos autores de manera independiente en base a las recomendaciones Prisma. Los criterios de selección fueron: artículos con cuestionarios validados en pacientes con DM2, en español o inglés y publicados hasta el 31 de noviembre de 2022.Se resumieron las pruebas de fiabilidad (consistencia interna, correlación ítem- total, coeficiente de correlación intraclase) y validez de los cuestionarios (validez de constructo, de criterio y de contenido). Resultados: 23 artículos formaron parte de la revisión. Se obtuvieron 10 cuestionarios de medida de la adherencia a los ADOs los cuales fueron validados en 14 países. El 8-item Self-Reported Medication Adherence Measure (MMAS-8) y el Adherence to Refills and Medications Scale (ARMS) fueron los más usados. Todos presentaron al menos una medida de validez y fiabilidad, siendo la validez de grupos conocidos la menos empleada. Conclusiones: Hay cuestionarios que presentan datos de validez o fiabilidad mejores que otros, pero deberá ser el profesional sanitario quien determine cuál se ajusta mejor a sus pacientes o a su estudio. (AU)

Introduction: adherence to oral antidiabetics (OADs) in patients with Type 2 Diabetes Mellitus (DM2) can affect disease control and therefore needs to be evaluated. For this, fast, simple, valid and reliable tools are needed. Thus, the objective of this review was to identify those validated questionnaires that are used to measure adherence to OADs in clinical practice. Method: a bibliographic search was carried out in the Medline, Scopus and LILACS databases. The selection of articles was performed by 2 authors independently based on Prisma recommendations. The selection criteria were: Articles with validated questionnaires in patients with DM2, in Spanish or English and published until 31 November 2022. Reliability tests (internal consistency, item-total correlation, intraclass correlation coefficient) and validity of the questionnaires (construct validity, criteria and content). Results: 23 articles were part of the review.10 questionnaires to measure adherence to the ODAs were obtained, which were validated in 14 countries. The 8-item Self-Reported Medication Adherence Measure (MMAS-8) and the Adherence to Refills and Medications Scale (ARMS) were the most used. All presented at least one measure of validity and reliability, the validity of known groups being the least used. Conclusions: There are questionnaires that present better validity or reliability data than others, but it should be the healthcare professional who determines which one best suits their patients or their study. (AU)

NAD pool as an antitumor target against cancer stem cells in head and neck cancer.

Head and neck squamous cell carcinoma (HNSCC) is a heterogeneous group of tumors that affect different anatomical locations. Despite this heterogeneity, HNSCC treatment depends on the anatomical location, TNM stage and resectability of the tumor. Classical chemotherapy is based on platinum-derived drugs (cisplatin, carboplatin and oxaliplatin), taxanes (docetaxel, paclitaxel) and 5-fluorouracil1. Despite advances in HNSCC treatment, the rate of tumor recurrence and patient mortality remain high. Therefore, the search for new prognostic identifiers and treatments targeting therapy-resistant tumor cells is vital. Our work demonstrates that there are different subgroups with high phenotypic plasticity within the CSC population in HNSCC. CD10, CD184, and CD166 may identify some of these CSC subpopulations with NAMPT as a common metabolic gene for the resilient cells of these subpopulations. We observed that NAMPT reduction causes a decrease in tumorigenic and stemness properties, migration capacity and CSC phenotype through NAD pool depletion. However, NAMPT-inhibited cells can acquire resistance by activating the NAPRT enzyme of the Preiss-Handler pathway. We observed that coadministration of the NAMPT inhibitor with the NAPRT inhibitor cooperated inhibiting tumor growth. The use of an NAPRT inhibitor as an adjuvant improved NAMPT inhibitor efficacy and reduced the dose and toxicity of these inhibitors. Therefore, it seems that the reduction in the NAD pool could have efficacy in tumor therapy. This was confirmed by in vitro assays supplying the cells with products of inhibited enzymes (NA, NMN or NAD) and restoring their tumorigenic and stemness properties. In conclusion, the coinhibition of NAMPT and NAPRT improved the efficacy of antitumor treatment, indicating that the reduction in the NAD pool is important to prevent tumor growth.

Neuronal Prosurvival Role of Ceramide Synthase 2 by Olidogendrocyte-to-Neuron Extracellular Vesicle Transfer.

Ageing is associated with notorious alterations in neurons, i.e., in gene expression, mitochondrial function, membrane degradation or intercellular communication. However, neurons live for the entire lifespan of the individual. One of the reasons why neurons remain functional in elderly people is survival mechanisms prevail over death mechanisms. While many signals are either pro-survival or pro-death, others can play both roles. Extracellular vesicles (EVs) can signal both pro-toxicity and survival. We used young and old animals, primary neuronal and oligodendrocyte cultures and neuroblastoma and oligodendrocytic lines. We analysed our samples using a combination of proteomics and artificial neural networks, biochemistry and immunofluorescence approaches. We found an age-dependent increase in ceramide synthase 2 (CerS2) in cortical EVs, expressed by oligodendrocytes. In addition, we show that CerS2 is present in neurons via the uptake of oligodendrocyte-derived EVs. Finally, we show that age-associated inflammation and metabolic stress favour CerS2 expression and that oligodendrocyte-derived EVs loaded with CerS2 lead to the expression of the antiapoptotic factor Bcl2 in inflammatory conditions. Our study shows that intercellular communication is altered in the ageing brain, which favours neuronal survival through the transfer of oligodendrocyte-derived EVs containing CerS2.

A retrospective cohort study comparing the social, familial and clinical characteristics of adolescents admitted to a French Emergency Department in suicidal crisis before and during the COVID-19 pandemic.

The social turmoil resulting from the COVID19 pandemic has come with an increase in the incidence of suicidal crises among adolescents and in particular an increase of suicidal attempts in most Western societies. Monthly prevalence of suicidal attempts in adolescents was doubled or tripled during winter 2021 in France. This study proposes to describe the youths in suicidal crisis admitted to a French hospital during the pandemic according to their sociodemographic, familial, and clinical characteristics as well as to compare them to the adolescents who were presenting the same symptoms the years preceding the pandemic. A retrospective cohort of 332 participants was divided in two groups. The adolescents admitted during the pandemic were more frequently girls, with less familial and personal history of psychiatric care, less depressive disorders, and they were more frequently referred to psychological outpatient care than to psychiatric outpatient care. These results strengthen the hypothesis that the increase in adolescents' suicidal crisis could be an expression of the social suffering more than a result of an increase in adolescents' psychiatric disorders.

Predictors of Word and Pseudoword Reading in Languages with Different Orthographic Consistency.

This study analyzes the impact of certain cognitive processes on word and pseudoword reading in languages with different orthographic consistency (Spanish and Arabic) in the first year of Primary Education. The study was conducted with a group of 113 pupils from Algeria and another group of 128 pupils from Ecuador, from a middle-class background and without any special education needs. The participants were assessed in terms of their reading ability of words and pseudowords, knowledge of letters, phonological awareness, rapid automatized naming, and phonological memory. Using a correlational design, descriptive-exploratory, bivariate, and hierarchical multivariate regressions were applied to the different measures of reading in each language. The findings show that knowledge of letters, phonological awareness, rapid automatized naming, and phonological memory contribute differently to the explanation of reading ability in each group at the start of compulsory schooling. These results have important implications for the teaching of reading skills and the prevention of specific learning disabilities, as well as the theory of reading acquisition.

Validación de un test de detección temprana de dificultades de la lectoescritura en Ecuador. Validación de un test de riesgo de dificultades / Validation of a test for early detection of literacy difficulties in Ecuador. Validation of a hardship risk test

Introducción: Las dificultades del aprendizaje son las alteraciones de mayor presencia en las aulas escolares y sus indicadores pueden diagnosticarse y prevenirse desde edades tempranas. El objetivo de esta investigación fue validar el Test para la detección temprana de las dificultades en el aprendizaje de la lectura y escritura. Métodos: El enfoque de la investigación fue cuantitativo, descriptivo y de corte transversal. Se utilizó la validez de constructo acorde con la propuesta original del test y de fiabilidad a través del Alpha de Cronbach en una muestra de 501 niños ecuatorianos de cuatro años. Resultados: La validación del instrumento evidencia una moderada correlación entre las sub-tareas y alta correlación entre las sub-tareas y el puntaje total. La fiabilidad es buena, α= 0.71, muy próxima a la de la población española α= 0.73. Por lo que, la prueba puede ser utilizada en el contexto ecuatoriano en su versión original, adecuando en las instrucciones dos palabras a la realidad lingüística del país y para la calificación los puntos de corte de dificultad. Conclusión: Considerando su valor y fácil aplicación se recomienda el uso de la prueba de lectura en contextos educativos y de salud.

Introduction: Learning difficulties are the alterations with the most significant presence in school classrooms, and their indicators can be diagnosed and prevented early. This research aimed to validate the test for the early detection of difficulties in learning to read and write. Methods: The research approach was quantitative, descriptive, and cross-sectional. Construct validity was used according to the original proposal of the test and reliability through Cronbach's alpha in a sample of 501 four-year-old Ecuadorian children. Results: The validation of the instrument shows a moderate correlation between the subtasks and a high correlation between the subtasks and the total score. The reliability is good, α = 0.71, very close to that of the Spanish population α = 0.73. Therefore, the test can be used in the Ecuadorian context in its original version, adapting two words in the instructions to the linguistic reality of the country and for the qualification of the cutoff points of difficulty. Conclusion: With the easy application of the "test of reading" in 4-year-old children, the authors recommended its application for the identification of dyslexia and phonological processing deficits in school children in Ecuador. The reading test's validity allows its application at a regional level.

Common Genetic Variation and Age of Onset of Anorexia Nervosa.

Background: Genetics and biology may influence the age of onset of anorexia nervosa (AN). The aims of this study were to determine whether common genetic variation contributes to age of onset of AN and to investigate the genetic associations between age of onset of AN and age at menarche. Methods: A secondary analysis of the Psychiatric Genomics Consortium genome-wide association study (GWAS) of AN was performed, which included 9335 cases and 31,981 screened controls, all from European ancestries. We conducted GWASs of age of onset, early-onset AN (<13 years), and typical-onset AN, and genetic correlation, genetic risk score, and Mendelian randomization analyses. Results: Two loci were genome-wide significant in the typical-onset AN GWAS. Heritability estimates (single nucleotide polymorphism-h 2) were 0.01-0.04 for age of onset, 0.16-0.25 for early-onset AN, and 0.17-0.25 for typical-onset AN. Early- and typical-onset AN showed distinct genetic correlation patterns with putative risk factors for AN. Specifically, early-onset AN was significantly genetically correlated with younger age at menarche, and typical-onset AN was significantly negatively genetically correlated with anthropometric traits. Genetic risk scores for age of onset and early-onset AN estimated from independent GWASs significantly predicted age of onset. Mendelian randomization analysis suggested a causal link between younger age at menarche and early-onset AN. Conclusions: Our results provide evidence consistent with a common variant genetic basis for age of onset and implicate biological pathways regulating menarche and reproduction.

Protocol to desensitize human and murine mast cells after polyclonal IgE sensitization.

In this protocol, we provide detailed instructions to desensitize human and murine mast cells (MCs) after polyclonal IgE sensitization. Moreover, we specify the steps for MC degranulation assessment after desensitization, measuring CD63 and CD107a expression by flow cytometry and ß-hexosaminidase activity. Desensitized MCs can be used directly for co-culture with other cell types, immunofluorescence, live imaging, and omics approaches. For complete details on the use and execution of this protocol, please refer to López-Sanz et al. (2022).

Impact of environmental conditions and gut microbiota on the in vitro germination and growth of Clostridioides difficile.

Clostridioides difficile is a spore-forming anaerobic Gram-positive bacterium responsible for a broad spectrum of intestinal symptoms and healthcare-associated diarrhoea. The hypothesis of this work was that different in vitro conditions, notably pH and human faecal microbiota composition, impact the germination and/or the growth of C. difficile. This study aimed to correlate growth kinetics of the bacterium with these two physiochemical parameters by using a static in vitro model. To better understand the initial gut colonisation, several growth curve assays were carried out to monitor the behaviour of the spores and vegetative forms of C. difficile strain 078 under different conditions mimicking the gut environment. When the faeces were added, no spore germination or growth was observed, but C. difficile spores germinated in vitro when the pH was maintained between 6.6 and 6.9 for four different faeces donors. The evolution of microbiota studied by 16S rDNA profiling showed high proportions of Enterobacteriaceae and E. coli/Shigella when C. difficile grew, regardless of the inoculated faeces. This model helped us to understand that the germination and growth of C. difficile are strongly pH dependent, and further research is needed to evaluate the potential impact of the gut microbiota composition on C. difficile.

Gut Microbiota Composition Associated with Clostridioides difficile Colonization and Infection.

Clostridioides difficile is an anaerobic Gram-positive and spore-forming bacterium. The majority of C. difficile strains produce two toxins, A and B, associated with the development of acute diarrhea and/or colitis. In this review, two situations are distinguished: C. difficile infection (CDI) and asymptomatic colonization (AC). The main objective of this review is to explore the available data related to the link between the gut microbiota and the development of CDI. The secondary aim is to provide more information on why some people colonized with toxigenic C. difficile develop an infection while others show no signs of disease. Several factors, such as the use of antibiotics and proton pump inhibitors, hospitalization, and age, predispose individuals to C. difficile colonization and/or C. difficile infection. The gut microbiota of people with AC showed decreased abundances of Prevotella, Alistipes, Bacteroides, Bifidobacterium, Dorea, Coprococcus, and Roseburia. The gut microbiota of people suffering from CDI showed reductions in the abundances of Lachnospiraceae, Ruminococcaceae, Blautia spp., Prevotella spp., Dialister spp., Bifidobacterium spp., Roseburia spp., Anaerostipes spp., Faecalibacterium spp. and Coprococcus spp., in comparison with healthy people. Furthermore, increases in the abundances of Enterococcaceae and Enterococcus were associated with C. difficile infection.