Alteraciones del metabolismo del hierro. Homocigosis H63D

Graus Morales, Javier, codir.La hemocromatosis hereditaria es el principal trastorno genético relacionado con el metabolismo del hierro. La mutación C282Y del gen HFE es el polimorfismo implicado más frecuentemente, si bien, la mutación H63D podría contribuir de forma independiente a la sobrecarga férrica. Dada la importancia de un diagnóstico precoz para prevenir el desarrollo de lesiones asociadas a la hemosiderosis y la elevada frecuencia de la mutación H63D en nuestra población, es fundamental determinar con mayor exactitud las implicaciones derivadas de la presencia de este polimorfismo. Como objetivo principal, nos propusimos analizar las implicaciones del polimorfismo H63D/H63D sobre el metabolismo del hierro. Como objetivos específicos, se analizaron las diferencias en la prevalencia de sobrecarga férrica entre los homocigotos H63D, los homocigotos C282Y y un grupo de sujetos no portadores de ninguna de estas 2 mutaciones, así como la influencia en estos grupos de otros factores asociados a hiperferritinemia. Se trata de un estudio observacional analítico que incluye 445 sujetos valorados en el Hospital Ramón y Cajal desde 1994 hasta Diciembre de 2013. El estudio genético se realizó mediante técnicas de Reacción en Cadena de la Polimerasa (PCR) en tiempo real. Se identificaron 192 pacientes homocigotos H63D, 109 homocigotos C282Y y 144 sujetos sin ninguna de las dos mutaciones. Los valores medios de ferritina e índice de saturación de la transferrina en los homocigotos H63D fueron 372.1± 385 mg/dl y 42.4± 19.1 %, respectivamente. Los sujetos de mayor edad presentaron concentraciones de ferritina más elevadas. La concentración media de AST, ALT, bilirrubina, GGT y hierro fueron normales. Treinta pacientes (15.62%) presentaban datos histológicos de sobrecarga férrica (medida mediante resonancia o biopsia), y 7 de estos, signos de cirrosis con hipertensión portal. En comparación con el grupo de homocigotos C282Y, el grupo H63D mostró menor concentración de ferritina, IST y menor porcentaje de sobrecarga parenquimatosa de hierro. Los sujetos sin mutaciones presentaban mayor concentración de ferritina que los homocigotos H63D, si bien, los porcentajes de IST, exceso parenquimatoso de hierro e hipertensión portal fueron similares. En el grupo sin mutaciones, se identificó una prevalencia significativamente mayor de factores asociados a hiperferritinemia secundaria, confirmando el origen no genético de la sobrecarga férrica en este grupo. Tras el análisis de los datos, nuestro estudio sugiere que los sujetos homocigotos H63D presentan concentraciones de ferritina elevadas, si bien, en ausencia de otros factores asociados a hiperferritinemia secundaria, este polimorfismo no determina una sobrecarga de hierro clínicamente significativa. En los pacientes homocigotos H63D que presentan datos analíticos o histológicos de sobrecarga férrica, debe descartarse en primer lugar la existencia de otros factores asociados a hiperferritinemia

Alteraciones del metabolismo del hierro. Homocigosis H63D

Graus Morales, Javier, codir.La hemocromatosis hereditaria es el principal trastorno genético relacionado con el metabolismo del hierro. La mutación C282Y del gen HFE es el polimorfismo implicado más frecuentemente, si bien, la mutación H63D podría contribuir de forma independiente a la sobrecarga férrica. Dada la importancia de un diagnóstico precoz para prevenir el desarrollo de lesiones asociadas a la hemosiderosis y la elevada frecuencia de la mutación H63D en nuestra población, es fundamental determinar con mayor exactitud las implicaciones derivadas de la presencia de este polimorfismo. Como objetivo principal, nos propusimos analizar las implicaciones del polimorfismo H63D/H63D sobre el metabolismo del hierro. Como objetivos específicos, se analizaron las diferencias en la prevalencia de sobrecarga férrica entre los homocigotos H63D, los homocigotos C282Y y un grupo de sujetos no portadores de ninguna de estas 2 mutaciones, así como la influencia en estos grupos de otros factores asociados a hiperferritinemia. Se trata de un estudio observacional analítico que incluye 445 sujetos valorados en el Hospital Ramón y Cajal desde 1994 hasta Diciembre de 2013. El estudio genético se realizó mediante técnicas de Reacción en Cadena de la Polimerasa (PCR) en tiempo real. Se identificaron 192 pacientes homocigotos H63D, 109 homocigotos C282Y y 144 sujetos sin ninguna de las dos mutaciones. Los valores medios de ferritina e índice de saturación de la transferrina en los homocigotos H63D fueron 372.1± 385 mg/dl y 42.4± 19.1 %, respectivamente. Los sujetos de mayor edad presentaron concentraciones de ferritina más elevadas. La concentración media de AST, ALT, bilirrubina, GGT y hierro fueron normales. Treinta pacientes (15.62%) presentaban datos histológicos de sobrecarga férrica (medida mediante resonancia o biopsia), y 7 de estos, signos de cirrosis con hipertensión portal. En comparación con el grupo de homocigotos C282Y, el grupo H63D mostró menor concentración de ferritina, IST y menor porcentaje de sobrecarga parenquimatosa de hierro. Los sujetos sin mutaciones presentaban mayor concentración de ferritina que los homocigotos H63D, si bien, los porcentajes de IST, exceso parenquimatoso de hierro e hipertensión portal fueron similares. En el grupo sin mutaciones, se identificó una prevalencia significativamente mayor de factores asociados a hiperferritinemia secundaria, confirmando el origen no genético de la sobrecarga férrica en este grupo. Tras el análisis de los datos, nuestro estudio sugiere que los sujetos homocigotos H63D presentan concentraciones de ferritina elevadas, si bien, en ausencia de otros factores asociados a hiperferritinemia secundaria, este polimorfismo no determina una sobrecarga de hierro clínicamente significativa. En los pacientes homocigotos H63D que presentan datos analíticos o histológicos de sobrecarga férrica, debe descartarse en primer lugar la existencia de otros factores asociados a hiperferritinemia

Un juego de cartas sencillo como metodología de aprendizaje de la química orgánica

Memoria ID-0173. Ayudas de la Universidad de Salamanca para la innovación docente, curso 2017-2018

Advances in Alzheimer’s Disease Research: Human Cerebral Organoids

Alzheimer’s disease (AD) is the main neurodegenerative disorder in old age, causing memory impairment and dependency. The histopathology of AD is characterized by the presence of amyloid plaques and neurofibrillary tangles formed by Aβ peptide and hyperphosphorylated Tau, respectively. There is still no cure or effective treatment for AD. This could be due, in part, to the lack of suitable research models since animal models do not recapitulate the full physiological complexity of the human brain. With the development of induced pluripotent stem cells (iPSCs), these limitations could be overcome. Even so, the bi-dimensional (2D) culture models still do not allow to recapitulate all types of brain cells and do not show a three-dimensional (3D) arrangement. Since obtaining 3D cultures called organoids, a new opportunity arises to overcome the limitations of previous models. Human Cerebral Organoids (hCOs) represent a pioneering model, in which part of the complexity of the human brain is present. For this reason, they are fast becoming a very remarkable model for the study of the evolution of the molecular and cellular pathology of AD. This review provides a brief overview of AD research, focusing on the most recent advances achieved through the development of stem cell and cerebral organoid technologyThe authors would like to thank to financing entities: i. State R+D+i Program Oriented to the Challenges of Society. Ministry of Science and Innovation (PID2021-126715OB-I00). ii. Strategic Action in Intramural Health (PI22/00055). iii. Ministry of Science and Innovation and Universities, within the program “R&D Projects «Retos Investigación» (RTI2018-101663-B-100).S

Hair cortisol level as a molecular biomarker in retinitis pigmentosa patients

Producción CientíficaPurpose: Retinitis pigmentosa (RP) patients commonly experience negative psychological states due to their progressive and unpredictable loss of vision and visual variations related to stress. The aim of this study was to examine hair cortisol concentrations (HCCs), which is usually associated with chronic stress, pretending to unveil possible associations between underlying psychological factors and disease severity in RP patients. Methods: Seventy-eight RP patients and 148 healthy controls were included in this study. A complete ophthalmological exam was performed in all patients to grade into severity disease groups. Perceived stress and trait-anxiety were measured by the State-Trait Anxiety Inventory (STAI) questionnaire. Results: Fifty-two (67%) patients had severe RP and 26 (33%) mild-moderate RP. Fifty-eight (58,9%) patients reported severely levels of stress and 18 (23.,1%) highly levels assessed by STAI questionnaire. RP patients exhibited higher HCCs (500.04 ± 120.99 pg/mg) than in controls (136.17 ± 60.51 pg/mg; p < 0.001). Severe RP patients had significant higher HCCs than mild-moderate patients differing in 274.27 pg/mg (p < 0.001). RP severity grade and perceived anxiety levels in the questionaries were not associated. Group differences were not affected by relevant covariates (age, grade of severity, stress status, and gender). Conclusions: HCC seems an effective biomarker associated with chronic stress in RP patients. This study shows that HCC in patients with RP are elevated compared to population-based controls, and association between HCC and RP severity was found. Future research is needed to characterize the effect of untreated negative psychological states on progression of the disease if any.Gerencia Regional de Salud de Castilla y León (GRS, 1932/A/19)Ministerio de Ciencia, Innovación y Universidades (grant PID2020-114585RA-I00

SINGLE-CELL sequencing workflow to study cellular composition and cell type specific expression profiles of human Cerebral Organoids

IBRO 11th World Congress of Neuroscience. Granada (Spain). 9-13 September 2023.Human cerebral organoid culture is a technology with immense potential in the areas of developmental neurobiology and neurodegeneration for example to study cell types, mechanisms involved, to discover of new biomarkers, to propose specific therapeutic strategies or to study the effects of compound-induced toxicity. Single-cell RNA sequencing (scRNA-seq) is a promising technology that will help to define the identity of the cerebral organoids and to understand cellular composition and cell type specific expression profiles. Standardization of workflows to do the scRNA-seq analysis is an important means to improve the use of this technology. We present the workflow and results of the scRNA-seq performed for cerebral organoids generated from the AND-2 cell line of human embryonic stem cells (hESCs). Dissociated cerebral organoid samples were loaded on the 10X Chromium and single cell libraries were prepared according to 10X Genomics standard procedures and sequenced on the Novaseq sequencer (Illumina).The data were checked and aligned to the GRCh38 human reference genome with CellRanger v6.0.2 and analyzed with Seurat v4.0. After quality filtering and data normalization with the SCTransform function, we performed Principal component analysis (PCA) using the highly variable genes, built a Shared Nearest Neighbor (SNN) graph using the Louvain method. To visualize data, Uniform Manifold Approximation and Projection (UMAP) dimensional reduction was performed. The identities of the cell clusters were assigned using the expression of genes specific of each cell type. We annotate in the AND2 cerebral organoids clusters for intermediate progenitor cells, astrocytes, oligodendrocyte precursor cells, excitatory neurons, inhibitory neurons, and mesodermal cells. We find also some cells in these organoids with expression of endothelial and microglial gene markers. Enrichment analysis of the highly variable differentially expressed genes (DEGs) was utilized to characterize the assigned cell types with Gene Ontology (GO), PanglaoDB and Cellmarker databases.S

Biologic Therapy for Moderate to Severe Psoriasis. Real-World Follow-up of Patients Who Initiated Biologic Therapy at Least 10 Years Ago

[Abstract] Introduction: The aim of this study was to evaluate response and drug survival of biologic therapy in patients with moderate to severe plaque-type psoriasis who initiated biologic therapy at least 10 years ago, in a real-world setting. Methods: This was an observational retrospective follow-up study that included patients with moderate to severe plaque-type psoriasis who initiated biologic therapy between October 2006 and December 2009. Efficacy was expressed as the percentage of patients achieving a 50, 75 and 90% reduction from baseline in the Psoriasis Area and Severity Index (PASI 50, PASI 75, PASI 90, respectively) every 3 months during the first year of therapy and then every 12 months up to the end of follow-up or withdrawal from the study. Results: A total of 56 patients were included in the study, representing 140 treatment lines (median 2, range 1-8); of these patients, 53 were still receiving biologic therapy at the end of the study. The mean duration of biologic therapy was 140.4 (range 47.6-175.4) months. Etanercept was used in 98.2% of patients, followed by efalizumab (42.9%), adalimumab (41.1%), ustekinumab (33.9%) and infliximab (16.1%). Treatment lines were switched in 62.1% of treatments: 24.3% due to secondary failure, 20.7% due to primary failure and 3.6% due to side effects. No patient treated with anti-interleukins had to discontinue treatment due to side effects. Ustekinumab had the highest drug survival. Conclusions: This study in the real-world-setting shows maintenance of long-term efficacy and safety of biologic therapy in patients with moderate to severe plaque psoriasis in daily practice who initiated biologic therapy 10 years ago.The journal’s Rapid Service Fee was paid for by Fundación Profesor Novoa Santos (A Coruña. Spain

Persistent Overactive Cytotoxic Immune Response in a Spanish Cohort of Individuals With Long-COVID: Identification of Diagnostic Biomarkers

Long-COVID is a new emerging syndrome worldwide that is characterized by the persistence of unresolved signs and symptoms of COVID-19 more than 4 weeks after the infection and even after more than 12 weeks. The underlying mechanisms for Long-COVID are still undefined, but a sustained inflammatory response caused by the persistence of SARS-CoV-2 in organ and tissue sanctuaries or resemblance with an autoimmune disease are within the most considered hypotheses. In this study, we analyzed the usefulness of several demographic, clinical, and immunological parameters as diagnostic biomarkers of Long-COVID in one cohort of Spanish individuals who presented signs and symptoms of this syndrome after 49 weeks post-infection, in comparison with individuals who recovered completely in the first 12 weeks after the infection. We determined that individuals with Long-COVID showed significantly increased levels of functional memory cells with high antiviral cytotoxic activity such as CD8+ TEMRA cells, CD8±TCRγδ+ cells, and NK cells with CD56+CD57+NKG2C+ phenotype. The persistence of these long-lasting cytotoxic populations was supported by enhanced levels of CD4+ Tregs and the expression of the exhaustion marker PD-1 on the surface of CD3+ T lymphocytes. With the use of these immune parameters and significant clinical features such as lethargy, pleuritic chest pain, and dermatological injuries, as well as demographic factors such as female gender and O+ blood type, a Random Forest algorithm predicted the assignment of the participants in the Long-COVID group with 100% accuracy. The definition of the most accurate diagnostic biomarkers could be helpful to detect the development of Long-COVID and to improve the clinical management of these patients.This work was supported by the Coordinated Research Activities at the National Center of Microbiology (CNM, Instituto de Salud Carlos III) (COV20_00679) to promote an integrated response against SARS-CoV-2 in Spain (Spanish Ministry of Science and Innovation), which is coordinated by Dr Inmaculada Casas (WHO National Influenza Center of the CNM); a generous donation provided by Chiesi España, S.A.U. (Barcelona, Spain); the Spanish Ministry of Science and Innovation (PID2019-110275RB-I00); and the Spanish AIDS Research Network RD16CIII/0002/0001 that is included in Acción Estratégica en Salud, Plan Nacional de Investigación Científica, Desarrollo e Innovación Tecnológica 2016-2020, Instituto de Salud Carlos III, European Region Development Fund (ERDF). The work of ML-H and SR-M is financed by NIH grant R01AI143567. The work of MT is supported by Instituto de Salud Carlos III (COV20_00679). The work of LV is supported by a pre-doctoral grant from Instituto de Salud Carlos III (FIS PI16CIII/00034-ISCIII-FEDER). The work of FR-M is financed by the Spanish Ministry of Science and Innovation (PID2019-110275RB-I00).S

Geotechnics for rockfall assessment in the volcanic island of Gran Canaria (Canary Islands, Spain)

The island of Gran Canaria (Canary Islands, Spain) is characterized by a large variability of volcanic rocks reflecting its volcanic evolution. The geological map provided by Geological Survey of Spain at 1:25.000 scale shows more than 109 different lithologies and it is too complex for environmental and engineering purposes. This work presents a simplified geotechnical map with a small number of classes grouping up units with similar geotechnical behaviours. The lithologies were grouped using about 350 rock samples, collected in the seven major islands of the Archipelago. The geotechnical map was used to model rockfall hazard in the entire island of Gran Canaria, where rockfalls are an important threat. The rockfall map was validated with 128 rockfall events along the GC-200 road, located in the NW sector of Gran Canaria. About 96% of the events occurred along sections of the road where the number of expected trajectories is high or moderate.This work was carried out in the framework of two projects funded by the European Commission, Directorate-General Humanitarian Aid and Civil Protection (ECHO): SAFETY (Sentinel-1 for geohazard prevention and forecasting. Ref. ECHO/SUB/2015/718679/Prev02) and U-GEOHAZ (Geohazard Impact Assessment for Urban Areas. Grant Agreement No. 783169). This work has been partly funded by the University of Alicante in the framework of Quality Improvement Grant of PhD Program in Materials, Structures and Soil Engineering: Sustainable Construction, Salvador de Madariaga Mobility Program from the Spanish Ministry of Science (PRX18/00020) and the Industrial PhD Project GEODRON (IND2017/AMB-7789). We also appreciate the contribution of the MACASTAB project (Ref.: MAC/3.5b/027). The laboratory tests were carried out in the Laboratories of Building and Public Works from the Canarian Government. The methodology is also developed in the framework of the RISKCOAST project (Ref: SOE3/P4/E0868) funded by the European Regional Development Fund - Interreg programme (3rd call for proposals)

O106 / #796 FEASIBILITY OF TRANSCUTANEOUS SPINAL CORD STIMULATION COMBINED WITH ROBOTIC-ASSISTED GAIT TRAINING (LOKOMAT) FOR GAIT REHABILITATION FOLLOWING INCOMPLETE SPINAL CORD INJURY. A CASE SERIES STUDY

Transcutaneous electrical spinal cord stimulation (tSCS) is a non-invasive technique for neuromodulation with therapeutic potential for motor rehabilitation following spinal cord injury (SCI). The aim of the present study was to analyze the feasibility of a program of 20 sessions of 30-Hz tSCS combined with robotic-assisted gait training in incomplete SCI. The results of the present work partially belong to a randomized clinical trial that is in progress