RESUMEN

The carboxy-terminus of the spliceosomal protein PRPF8, which regulates the RNA helicase Brr2, is a hotspot for mutations causing retinitis pigmentosa-type 13, with unclear role in human splicing and tissue-specificity mechanism. We used patient induced pluripotent stem cells-derived cells, carrying the heterozygous PRPF8 c.6926 A > C (p.H2309P) mutation to demonstrate retinal-specific endophenotypes comprising photoreceptor loss, apical-basal polarity and ciliary defects. Comprehensive molecular, transcriptomic, and proteomic analyses revealed a role of the PRPF8/Brr2 regulation in 5'-splice site (5'SS) selection by spliceosomes, for which disruption impaired alternative splicing and weak/suboptimal 5'SS selection, and enhanced cryptic splicing, predominantly in ciliary and retinal-specific transcripts. Altered splicing efficiency, nuclear speckles organisation, and PRPF8 interaction with U6 snRNA, caused accumulation of active spliceosomes and poly(A)+ mRNAs in unique splicing clusters located at the nuclear periphery of photoreceptors. Collectively these elucidate the role of PRPF8/Brr2 regulatory mechanisms in splicing and the molecular basis of retinal disease, informing therapeutic approaches.

Asunto(s)

Sitios de Empalme de ARN , Retinitis Pigmentosa , Empalmosomas , Humanos , Empalmosomas/genética , Empalmosomas/metabolismo , Proteómica , Empalme del ARN/genética , Empalme Alternativo/genética , ARN Nuclear Pequeño/genética , ARN Nuclear Pequeño/metabolismo , ARN Mensajero/metabolismo , Mutación , ADN Helicasas/metabolismo , Proteínas de Unión al ARN/metabolismo

RESUMEN

Molecular information on the early stages of human retinal development remains scarce due to limitations in obtaining early human eye samples. Pluripotent stem cell-derived retinal organoids (ROs) provide an unprecedented opportunity for studying early retinogenesis. Using a combination of single cell RNA-seq and spatial transcriptomics we present for the first-time a single cell spatiotemporal transcriptome of RO development. Our data demonstrate that ROs recapitulate key events of retinogenesis including optic vesicle/cup formation, presence of a putative ciliary margin zone, emergence of retinal progenitor cells and their orderly differentiation to retinal neurons. Combining the scRNA- with scATAC-seq data, we were able to reveal cell-type specific transcription factor binding motifs on accessible chromatin at each stage of organoid development, and to show that chromatin accessibility is highly correlated to the developing human retina, but with some differences in the temporal emergence and abundance of some of the retinal neurons.

RESUMEN

Nowadays, chronic kidney disease (CKD) prevalence keeps increasing worldwide. The management of these patients usually requires renal replacement therapy (RRT). However, the complexity of patients' profiles comprises a great challenge to overcome. During the last decades, CKD units have been developed to offer multidisciplinary and coordinated attention to patients, helping in the decision-making of the RRT. Nevertheless, there is a huge variability in the performance and organization of care practice, implying an existing necessity to homogenize the RRT modality chosen. We propose a test composed of two parts: one to be completed by the medical staff (to evaluate contraindications for the different RRT techniques) and another by the patient or nursing staff (to consider patients' preferences). In this sense, it would be possible to have a common and useful tool to complement patient education in RRT, as well as sharing decision-making in the ACKD units taking into account patient preferences.

Asunto(s)

Terapia de Reemplazo Renal Continuo , Insuficiencia Renal Crónica , Humanos , Diálisis Renal/métodos , Insuficiencia Renal Crónica/diagnóstico , Insuficiencia Renal Crónica/terapia , Terapia de Reemplazo Renal/métodos , Prevalencia

RESUMEN

Microglia are the primary resident immune cells in the retina. They regulate neuronal survival and synaptic pruning making them essential for normal development. Following injury, they mediate adaptive responses and under pathological conditions they can trigger neurodegeneration exacerbating the effect of a disease. Retinal organoids derived from human induced pluripotent stem cells (hiPSCs) are increasingly being used for a range of applications, including disease modelling, development of new therapies and in the study of retinogenesis. Despite many similarities to the retinas developed in vivo, they lack some key physiological features, including immune cells. We engineered an hiPSC co-culture system containing retinal organoids and microglia-like (iMG) cells and tested their retinal invasion capacity and function. We incorporated iMG into retinal organoids at 13 weeks and tested their effect on function and development at 15 and 22 weeks of differentiation. Our key findings showed that iMG cells were able to respond to endotoxin challenge in monocultures and when co-cultured with the organoids. We show that retinal organoids developed normally and retained their ability to generate spiking activity in response to light. Thus, this new co-culture immunocompetent in vitro retinal model provides a platform with greater relevance to the in vivo human retina.

Asunto(s)

Células Madre Pluripotentes Inducidas , Humanos , Microglía , Retina , Organoides , Diferenciación Celular

RESUMEN

Retinal drug toxicity screening is essential for the development of safe treatment strategies for a large number of diseases. To this end, retinal organoids derived from human pluripotent stem cells (hPSCs) provide a suitable screening platform due to their similarity to the human retina and the ease of generation in large-scale formats. In this study, two hPSC cell lines were differentiated to retinal organoids, which comprised all key retinal cell types in multiple nuclear and synaptic layers. Single-cell RNA-Seq of retinal organoids indicated the maintenance of retinal ganglion cells and development of bipolar cells: both cell types segregated into several subtypes. Ketorolac, digoxin, thioridazine, sildenafil, ethanol, and methanol were selected as key compounds to screen on retinal organoids because of their well-known retinal toxicity profile described in the literature. Exposure of the hPSC-derived retinal organoids to digoxin, thioridazine, and sildenafil resulted in photoreceptor cell death, while digoxin and thioridazine additionally affected all other cell types, including Müller glia cells. All drug treatments caused activation of astrocytes, indicated by dendrites sprouting into neuroepithelium. The ability to respond to light was preserved in organoids although the number of responsive retinal ganglion cells decreased after drug exposure. These data indicate similar drug effects in organoids to those reported in in vivo models and/or in humans, thus providing the first robust experimental evidence of their suitability for toxicological studies.

Asunto(s)

Células Madre Pluripotentes Inducidas , Organoides , Diferenciación Celular , Digoxina/metabolismo , Digoxina/farmacología , Humanos , Retina/metabolismo , Citrato de Sildenafil/metabolismo , Citrato de Sildenafil/farmacología , Tioridazina/metabolismo , Tioridazina/farmacología

RESUMEN

INTRODUCTION: Mutations in pre-mRNA processing factor 31 (PRPF31), a core protein of the spliceosomal tri-snRNP complex, cause autosomal-dominant retinitis pigmentosa (adRP). It has remained an enigma why mutations in ubiquitously expressed tri-snRNP proteins result in retina-specific disorders, and so far, the underlying mechanism of splicing factors-related RP is poorly understood. METHODS: We used the induced pluripotent stem cell (iPSC) technology to generate retinal organoids and RPE models from four patients with severe and very severe PRPF31-adRP, unaffected individuals and a CRISPR/Cas9 isogenic control. RESULTS: To fully assess the impacts of PRPF31 mutations, quantitative proteomics analyses of retinal organoids and RPE cells were carried out showing RNA splicing, autophagy and lysosome, unfolded protein response (UPR) and visual cycle-related pathways to be significantly affected. Strikingly, the patient-derived RPE and retinal cells were characterised by the presence of large amounts of cytoplasmic aggregates containing the mutant PRPF31 and misfolded, ubiquitin-conjugated proteins including key visual cycle and other RP-linked tri-snRNP proteins, which accumulated progressively with time. The mutant PRPF31 variant was not incorporated into splicing complexes, but reduction of PRPF31 wild-type levels led to tri-snRNP assembly defects in Cajal bodies of PRPF31 patient retinal cells, altered morphology of nuclear speckles and reduced formation of active spliceosomes giving rise to global splicing dysregulation. Moreover, the impaired waste disposal mechanisms further exacerbated aggregate formation, and targeting these by activating the autophagy pathway using Rapamycin reduced cytoplasmic aggregates, leading to improved cell survival. CONCLUSIONS: Our data demonstrate that it is the progressive aggregate accumulation that overburdens the waste disposal machinery rather than direct PRPF31-initiated mis-splicing, and thus relieving the RPE cells from insoluble cytoplasmic aggregates presents a novel therapeutic strategy that can be combined with gene therapy studies to fully restore RPE and retinal cell function in PRPF31-adRP patients.

Asunto(s)

Autofagia , Proteínas del Ojo , Células Madre Pluripotentes Inducidas , Agregado de Proteínas , Retinitis Pigmentosa , Proteínas del Ojo/genética , Proteínas del Ojo/metabolismo , Humanos , Células Madre Pluripotentes Inducidas/metabolismo , Epitelio Pigmentado de la Retina/citología , Epitelio Pigmentado de la Retina/metabolismo , Retinitis Pigmentosa/genética , Retinitis Pigmentosa/metabolismo , Ribonucleoproteínas Nucleares Pequeñas

RESUMEN

BACKGROUND: Brain-derived neurotrophic factor (BDNF) plays an important role in brain development by regulating multiple pathways within the central nervous system. In the Human Biomonitoring for Europe Project (HBM4EU), this neurotrophin is being implemented as a novel effect biomarker to evaluate the potential threats of environmental chemicals on neurodevelopment. OBJECTIVES: To explore the relationships among exposure to environmental metals, BDNF biomarkers at two levels of biological complexity, and behavioral function in adolescent males. METHODS: Data were gathered from 125 adolescents on: spot urine sample total concentrations of the neurotoxic metal(oid)s arsenic (As), cadmium (Cd), mercury (Hg), and lead (Pb); serum BDNF protein concentrations; and concurrent behavioral functioning according to the Child Behavior Check List (CBCL/6-18). In 113 of the participants, information was also collected on blood BDNF DNA methylation at six CpGs. Associations were evaluated by multivariate linear regression analysis adjusted for confounders. RESULTS: As, Cd, Hg, and Pb were detected in 100%, 98.5%, 97.0%, and 89.5% of urine samples, respectively. Median serum BDNF concentration was 32.6 ng/mL, and total percentage of BDNF gene methylation was 3.8%. In the adjusted models, urinary As was non-linearly associated with more internalizing problems and Cd with more externalizing behaviors. The percentage BDNF DNA methylation at CPGs #5 and the mean percentage CpG methylation increased across As tertiles (p-trend = 0.04 and 0.03, respectively), while 2nd tertile and 3rd tertile of Cd concentrations were associated with lower serum BDNF and higher CpG3 methylation percentage. Additionally, when BDNF was categorized in tertiles, serum BDNF at the 3rd tertile was associated with fewer behavioral problems, particularly withdrawn (p-trend = 0.04), social problems (p-trend = 0.12), and thought problems (p-trend = 0.04). CONCLUSION: Exposure to As and Cd was associated with BDNF gene DNA methylation BDNF gene and serum BDNF, respectively. Associations with DNA methylation may be attributable to a higher variability over time in circulating BDNF concentrations than in the methylation status of this gene. Caution should be taken when interpreting the results relating postnatal Pb and Hg to behavioral functioning. Further studies are needed to verify these findings.

Asunto(s)

Conducta del Adolescente , Factor Neurotrófico Derivado del Encéfalo , Exposición a Riesgos Ambientales , Metales , Adolescente , Arsénico , Factor Neurotrófico Derivado del Encéfalo/genética , Metilación de ADN , Humanos , Masculino , Mercurio , Metales/orina

RESUMEN

BACKGROUND AND PURPOSE: Primary lateral sclerosis (PLS) is a progressive upper motor neuron (UMN) disorder. It is debated whether PLS is part of the amyotrophic lateral sclerosis (ALS) spectrum, or a syndrome encompassing different neurodegenerative diseases. Recently, new diagnostic criteria for PLS have been proposed. We describe four patients of two pedigrees, meeting definite PLS criteria and harboring two different mutations in presenilin 1 (PSEN1). METHODS: Patients underwent neurological and neuropsychological examination, MRI, 18F-fluorodeoxyglucose positron emission tomography (FDG-PET), amyloid-related biomarkers, and next-generation sequencing (NGS) testing. RESULTS: Four patients, aged 25-45 years old, presented with a progressive UMN syndrome meeting clinical criteria of definite PLS. Cognitive symptoms and signs were mild or absent during the first year of the disease but appeared or progressed later in the disease course. Brain MRI showed microbleeds in two siblings, but iron-related hypointensities in the motor cortex were absent. Brain FDG-PET showed variable areas of hypometabolism, including the motor cortex and frontotemporal lobes. Amyloid deposition was confirmed with either cerebrospinal fluid (CSF) or imaging biomarkers. Two heterozygous likely pathogenic mutations in PSEN1 (p.Pro88Leu and p.Leu166Pro) were found in the NGS testing. CONCLUSION: Clinically defined PLS is a syndrome encompassing different neurodegenerative diseases. The NGS testing should be part of the diagnostic workup in patients with PLS, at least in those with red flags, such as early-onset, cognitive impairment, and/or family history of neurodegenerative diseases.

RESUMEN

As one of the primary points of entry of xenobiotic substances and infectious agents into the body, the lungs are subject to a range of dysfunctions and diseases that together account for a significant number of patient deaths. In view of this, there is an outstanding need for in vitro systems in which to assess the impact of both infectious agents and xenobiotic substances of the lungs. To address this issue, we have developed a protocol to generate airway epithelial basal-like cells from induced pluripotent stem cells, which simplifies the manufacture of cellular models of the human upper airways. Basal-like cells generated in this study were cultured on transwell inserts to allow formation of a confluent monolayer and then exposed to an air-liquid interface to induce differentiation into a pseudostratified epithelial construct with a marked similarity to the upper airway epithelium in vivo. These constructs contain the component cell types required of an epithelial model system, produce mucus and functional cilia, and can support SARS-CoV-2 infection/replication and the secretion of cytokines in a manner similar to that of in vivo airways. This method offers a readily accessible and highly scalable protocol for the manufacture of upper airway models that could find applications in development of therapies for respiratory viral infections and the assessment of drug toxicity on the human lungs.

Asunto(s)

COVID-19/patología , COVID-19/virología , Células Madre Pluripotentes Inducidas/patología , Pulmón/patología , Pulmón/virología , Modelos Biológicos , SARS-CoV-2/fisiología , Línea Celular , Citocinas/metabolismo , Células Epiteliales/patología , Células Epiteliales/virología , Humanos , Mediadores de Inflamación/metabolismo , Replicación Viral/fisiología

RESUMEN

PURPOSE: Single cell (sc) analyses of key embryonic, fetal and adult stages were performed to generate a comprehensive single cell atlas of all the corneal and adjacent conjunctival cell types from development to adulthood. METHODS: Four human adult and seventeen embryonic and fetal corneas from 10 to 21 post conception week (PCW) specimens were dissociated to single cells and subjected to scRNA- and/or ATAC-Seq using the 10x Genomics platform. These were embedded using Uniform Manifold Approximation and Projection (UMAP) and clustered using Seurat graph-based clustering. Cluster identification was performed based on marker gene expression, bioinformatic data mining and immunofluorescence (IF) analysis. RNA interference, IF, colony forming efficiency and clonal assays were performed on cultured limbal epithelial cells (LECs). RESULTS: scRNA-Seq analysis of 21,343 cells from four adult human corneas and adjacent conjunctivas revealed the presence of 21 cell clusters, representing the progenitor and differentiated cells in all layers of cornea and conjunctiva as well as immune cells, melanocytes, fibroblasts, and blood/lymphatic vessels. A small cell cluster with high expression of limbal progenitor cell (LPC) markers was identified and shown via pseudotime analysis to give rise to five other cell types representing all the subtypes of differentiated limbal and corneal epithelial cells. A novel putative LPCs surface marker, GPHA2, expressed on the surface of 0.41% ± 0.21 of the cultured LECs, was identified, based on predominant expression in the limbal crypts of adult and developing cornea and RNAi validation in cultured LECs. Combining scRNA- and ATAC-Seq analyses, we identified multiple upstream regulators for LPCs and demonstrated a close interaction between the immune cells and limbal progenitor cells. RNA-Seq analysis indicated the loss of GPHA2 expression and acquisition of proliferative limbal basal epithelial cell markers during ex vivo LEC expansion, independently of the culture method used. Extending the single cell analyses to keratoconus, we were able to reveal activation of collagenase in the corneal stroma and a reduced pool of limbal suprabasal cells as two key changes underlying the disease phenotype. Single cell RNA-Seq of 89,897 cells obtained from embryonic and fetal cornea indicated that during development, the conjunctival epithelium is the first to be specified from the ocular surface epithelium, followed by the corneal epithelium and the establishment of LPCs, which predate the formation of limbal niche by a few weeks. CONCLUSIONS: Our scRNA-and ATAC-Seq data of developing and adult cornea in steady state and disease conditions provide a unique resource for defining genes/pathways that can lead to improvement in ex vivo LPCs expansion, stem cell differentiation methods and better understanding and treatment of ocular surface disorders.

Asunto(s)

Epitelio Corneal , Limbo de la Córnea , Adulto , Diferenciación Celular , Células Cultivadas , Córnea , Células Epiteliales , Humanos , Células Madre

RESUMEN

Development of the retina is regulated by growth factors, such as insulin-like growth factors 1 and 2 (IGF-1/2), which coordinate proliferation, differentiation, and maturation of the neuroepithelial precursors cells. In the circulation, IGF-1/2 are transported by the insulin growth factor binding proteins (IGFBPs) family members. IGFBPs can impact positively and negatively on IGF-1, by making it available or sequestering IGF-1 to or from its receptor. In this study, we investigated the expression of IGFBPs and their role in the generation of human retinal organoids from human pluripotent stem cells, showing a dynamic expression pattern suggestive of different IGFBPs being used in a stage-specific manner to mediate IGF-1 functions. Our data show that IGF-1 addition to culture media facilitated the generation of retinal organoids displaying the typical laminated structure and photoreceptor maturation. The organoids cultured in the absence of IGF-1, lacked the typical laminated structure at the early stages of differentiation and contained significantly less photoreceptors and more retinal ganglion cells at the later stages of differentiation, confirming the positive effects of IGF-1 on retinal lamination and photoreceptor development. The organoids cultured with the IGFBP inhibitor (NBI-31772) and IGF-1 showed lack of retinal lamination at the early stages of differentiation, an increased propensity to generate horizontal cells at mid-stages of differentiation and reduced photoreceptor development at the later stages of differentiation. Together these data suggest that IGFBPs enable IGF-1's role in retinal lamination and photoreceptor development in a stage-specific manner.

Asunto(s)

Proteínas de Unión a Factor de Crecimiento Similar a la Insulina/genética , Factor II del Crecimiento Similar a la Insulina/genética , Factor I del Crecimiento Similar a la Insulina/genética , Organoides/metabolismo , Células Fotorreceptoras de Vertebrados/metabolismo , Células Madre Pluripotentes/metabolismo , Catecoles/farmacología , Diferenciación Celular/efectos de los fármacos , Proteína 3 Similar a ELAV/genética , Proteína 3 Similar a ELAV/metabolismo , Proteína 4 Similar a ELAV/genética , Proteína 4 Similar a ELAV/metabolismo , Regulación de la Expresión Génica , Proteínas de Homeodominio/genética , Proteínas de Homeodominio/metabolismo , Humanos , Proteínas de Unión a Factor de Crecimiento Similar a la Insulina/antagonistas & inhibidores , Proteínas de Unión a Factor de Crecimiento Similar a la Insulina/metabolismo , Factor I del Crecimiento Similar a la Insulina/metabolismo , Factor I del Crecimiento Similar a la Insulina/farmacología , Factor II del Crecimiento Similar a la Insulina/metabolismo , Isoquinolinas/farmacología , Antígeno Ki-67/genética , Antígeno Ki-67/metabolismo , Proteínas de Neoplasias/genética , Proteínas de Neoplasias/metabolismo , Organoides/citología , Organoides/efectos de los fármacos , Células Fotorreceptoras de Vertebrados/citología , Células Fotorreceptoras de Vertebrados/efectos de los fármacos , Células Madre Pluripotentes/citología , Células Madre Pluripotentes/efectos de los fármacos , Recoverina/genética , Recoverina/metabolismo , Transducción de Señal , Transactivadores/genética , Transactivadores/metabolismo , Factores de Transcripción/genética , Factores de Transcripción/metabolismo , gamma-Sinucleína/genética , gamma-Sinucleína/metabolismo

RESUMEN

A 22-year-old male was referred for headaches, hearing impairment, and right eye scotoma. Branch retinal artery occlusion was revealed during the ophthalmological examination. Susac syndrome was suspected due to the symptoms described and the absence of cardiovascular risk factors. An extensive ophthalmological examination, including multimodal imaging was carried out, which is of special interest as it is considered to be a rare syndrome.

RESUMEN

Age-related macular degeneration (AMD) is a multifactorial disease, which is characterized by loss of central vision, affecting one in three people by the age of 75. The Y402H polymorphism in the complement factor H (CFH) gene significantly increases the risk of AMD. We show that Y402H-AMD-patient-specific retinal pigment epithelium (RPE) cells are characterized by a significant reduction in the number of melanosomes, an increased number of swollen lysosome-like-vesicles with fragile membranes, Cathepsin D leakage into drusen-like deposits and reduced lysosomal function. The turnover of C3 is increased significantly in high-risk RPE cells, resulting in higher internalization and deposition of the terminal complement complex C5b-9 at the lysosomes. Inhibition of C3 processing via the compstatin analogue Cp40 reverses the disease phenotypes by relieving the lysosomes of their overburden and restoring their function. These findings suggest that modulation of the complement system represents a useful therapeutic approach for AMD patients associated with complement dysregulation.

Asunto(s)

Degeneración Macular/terapia , Epitelio Pigmentado de la Retina/metabolismo , Femenino , Humanos , Lisosomas/metabolismo , Degeneración Macular/patología , Masculino

RESUMEN

Tools to assess diet in a reliable and efficient way are needed, particularly in children and adolescents. In this study, we assess the reproducibility and validity of a semiquantitative food frequency questionnaire (FFQ) among adolescents in Spain. We analyzed data of 51 male adolescents aged 15-17 years from a prospective birth cohort study. Participants answered the FFQ twice in a self-administered way over a 12-month period. Reproducibility was assessed by comparing nutrient and food intakes from the FFQs, and validity by comparing nutrient intakes from the average of two FFQs and the average of two 24-Hour Dietary Recalls obtained in the period. Pearson correlation coefficients were calculated. The average of reproducibility correlation coefficients for food group intakes was 0.33, with the highest correlation for vegetable intake (r = 0.81); and the average for nutrient intake was 0.32, with the highest coefficients for α- and ß-carotene (r = 0.65). Validity correlation coefficients ranged from 0.07 for carbohydrates to 0.53 for dietary fiber. The average of the validity correlation coefficients was r = 0.32. This study suggests that our FFQ may be a useful tool for assessing dietary intake of most nutrient and food groups among Spanish male adolescents in a self-administered way, despite reproducibility and, particularly validity, being low for some nutrients and food groups.

Asunto(s)

Fenómenos Fisiológicos Nutricionales de los Adolescentes/fisiología , Ingestión de Alimentos/fisiología , Conducta Alimentaria/fisiología , Evaluación Nutricional , Encuestas Nutricionales/métodos , Encuestas y Cuestionarios , Adolescente , Estudios de Cohortes , Registros de Dieta , Humanos , Masculino , Reproducibilidad de los Resultados , España , Factores de Tiempo , Verduras

RESUMEN

Effective classroom management is a critical teaching skill and a key concern for educators. Disruptive behaviors disturb effective classroom management and can influence school satisfaction if the teacher does not have the competencies to control them. Two objectives were set in this work: to understand the differences that exist in school satisfaction, disruptive behaviors, and teaching competencies according to the gender of the students; and to analyze school satisfaction and disruptive student behaviors based on perceived teaching competence. A non-probabilistic and convenience sample selection process was employed, based on the subjects that we were able to access. 758 students participated (male = 45.8%) from seven public secondary schools in the Murcia Region (Spain). The age range was between 13 and 18 years (M = 15.22; DT = 1.27). A questionnaire composed of the following scales was used: Competencies Evaluation Scale for Teachers in Physical Education, School Satisfaction and Disruptive Behaviors in Physical Education. Mixed Linear Models performed with the SPSS v.23 was used for statistical analyses. The results revealed statistically significant differences based on gender and physical education teaching competencies. In conclusion, the study highlights that physical education teacher skills influence disruptive behaviors in the classroom, and that this is also related to school satisfaction. Furthermore, it highlights that boys showed higher levels of negative behaviors than girls.

RESUMEN

OBJECTIVE: To examine the association of urinary concentrations of arsenic (As), cadmium (Cd), mercury (Hg), nickel (Ni), lead (Pb), manganese (Mn), and chromium (Cr) with blood pressure (BP) and serum hormone levels in male adolescents. METHODS: Participants were selected from the INMA (Environment and Childhood)-Granada cohort at their follow-up visit when aged 15-17 years. Metal concentrations were measured in urine samples using inductively coupled plasma mass spectrometry. Outcomes were BP measurements (systolic, diastolic, and pulse pressure) recorded during the visit and concurrent serum levels of thyroid hormones, sex hormones, and adrenal hormones. Associations were assessed by regression analysis in a sub-sample of 133 boys with available data on urinary metals, outcomes, and relevant covariates. RESULTS: Models simultaneously adjusted for all metals and other potential confounders showed that urinary As and Cd were both associated with slight elevations in systolic BP (0.70 mmHg, 95%CI = 0.11; 1.29 and 1.47, 95%CI = 0.30; 2.63, respectively, per each 50% increase in metal concentrations), and urinary As was also associated with an increased risk of elevated systolic BP (≥120 mmHg) (OR = 1.28, 95%CI = 1.04; 1.56). The presence of detectable levels of 4 and 5 versus 2-3 non-essential metals (As, Cd, Hg, Ni, Pb) per boy was associated with elevations in systolic BP of 5.84 mmHg (95%CI = 0.40; 11.3) and 7.01 mmHg (95%CI = 1.01; 13.0), respectively (p-trend = 0.05). Significant associations were also found between Hg and increased testosterone and luteinizing hormone (LH) and decreased thyroid-stimulating hormone (TSH); between the combination of As and Hg and increased LH and insulin-like growth factor 1; between Cr and decreased TSH; and between Cd and increased adrenocorticotropic hormone. CONCLUSIONS: These findings suggest that combined exposure to toxic metals, especially As and Cd, may contribute to BP elevation in male adolescents and that exposure to Hg, As, Cd, and Cr may affect their hormone levels.

Asunto(s)

Arsénico , Presión Sanguínea , Mercurio , Metales Pesados , Adolescente , Cadmio , Humanos , Masculino , Metales Pesados/toxicidad , Metales Pesados/orina

RESUMEN

Objective: To analyze those factors contributing to the diagnostic delay in ALS. Methods: Consecutive ALS patients were categorized as those studied in departmental hospitals and those studied in a referral ALS center. Demographic and clinical variables, together with data of the diagnostic pathway were collected. Multivariable models were used to assess their effect in the time between symptoms onset and the first neurologist visit (time symptoms-neurologist), in the time between the first neurologist visit and the diagnosis (time neurologist-diagnosis) and in the diagnostic delay. Results: 166 ALS patients with a median diagnostic delay of 11.53 months (IQR: 6.68, 15.23) were included. The median diagnostic delay was 8.57 months (5.16, 11.61) in the referral center vs. 12.08 months (6.87, 16.8) in departmental centers. Bulbar onset, fast progression rate, upper motor neuron predominant phenotype and an early referral to the neurologist were associated with a shorter time between symptoms-neurologist. Being studied in a referral center was associated with a shorter time between neurologist-diagnosis. Comorbidities, familial ALS, bulbar onset, early referral to the neurologist and being studied in a referral center were associated with a shorter diagnostic delay. For patients studied in departmental hospitals, fast progression rate was also strongly associated with a shorter time between neurologist-diagnosis and diagnostic delay. Conclusion: Unmodifiable factors (comorbidities, familial ALS, bulbar onset, and progression rate) as well as modifiable factors (early referral to the neurologist and the evaluation in an ALS referral center) have an independent effect in the diagnostic delay. The universalization of ALS Units is probably the most efficient measure to reduce the diagnostic delay.

RESUMEN

Currently, disruptive and aggressive behaviours of a physical and verbal nature are a reality among adolescent students and a concern in the educational context. Therefore, the main objective of this research was to analyse the effects of perceived teaching competence, motivation and basic psychological needs on disruptive behaviours in secondary school PE students. The sample was composed of 758 adolescent students from seven public secondary schools. The following instruments adapted to physical education were used: The Disruptive Behaviours Questionnaire, The Evaluation of Teaching Competencies Scale, The Sport Motivation Scale, and The Basic Psychological Needs Scale. Multilevel regression models with the MIXED procedure were performed for data analysis. The results show that misbehaviour is more likely among male students and that disruptive behaviours decrease when a teacher is perceived as competent. Students with greater self-determined motivation are more likely to exhibit fewer behaviours related to low engagement and irresponsibility while amotivation increases the different disruptive behaviours in the classroom. In conclusion, it is proposed that educators work in line with the students' needs by responding to their interests and that this will increase self-determined motivation.

Asunto(s)

Necesidades y Demandas de Servicios de Salud , Motivación , Educación y Entrenamiento Físico , Problema de Conducta , Instituciones Académicas , Estudiantes/psicología , Adolescente , Femenino , Humanos , Masculino , Encuestas y Cuestionarios

RESUMEN

The objectives of this work were two-fold: Firstly, to identify the profiles of disruptive behaviours and motivation in secondary school physical education students using cluster analysis; and secondly, to analyse the interaction of the profiles with school satisfaction and perceived teaching competence. A group of 758 secondary school students (54.2% female) between the ages of 13 and 18 (M = 15.22, DT = 1.27) participated in the study by responding to the following scales: The Disruptive Behaviours in Physical Education Questionnaire, The School Satisfaction Scale, The Sport Motivation Scale adapted to Physical Education, and the Evaluation of Teaching Competencies Scale in Physical Education. The cluster analysis established two distinct profiles: High levels of disruptive behaviours and low levels of disruptive behaviours. The results showed that the students with the high disruptive behaviours profile were mostly boys, having low levels of intrinsic motivation and high levels of amotivation and misbehaviour in the classroom. In contrast, those students with the low disruptive behaviours profile were mostly girls, having the highest levels of intrinsic motivation and the lowest levels in all the disruptive behaviours. It was shown that students exhibiting the worse classroom behaviours were more bored in school, while those students with better behaviour perceived greater teaching competence.

Asunto(s)

Trastornos de la Conducta Infantil/psicología , Motivación , Educación y Entrenamiento Físico , Problema de Conducta , Instituciones Académicas , Adolescente , Análisis por Conglomerados , Femenino , Humanos , Masculino , Satisfacción Personal , Examen Físico , Encuestas y Cuestionarios

RESUMEN

PRPF31 gene codes for a ubiquitously expressed splicing factor but mutations affect exclusively the retina, producing the progressive death of photoreceptor cells. We have identified a novel PRPF31 mutation in a patient with autosomal dominant retinitis pigmentosa. A blood sample was obtained and mononuclear cells were reprogrammed using the non-integrative Sendai virus to generate the cell line CABi001-A. The iPSC line has been characterized for pluripotency and differentiation capacity and will be differentiated toward photoreceptors and retinal pigment epithelium cells to study the molecular mechanism of the disease and test possible therapeutic strategies.

Asunto(s)

Línea Celular , Proteínas del Ojo/genética , Células Madre Pluripotentes Inducidas , Retinitis Pigmentosa/genética , Anciano , Diferenciación Celular , Técnicas de Reprogramación Celular , Femenino , Heterocigoto , Humanos , Cariotipo , Leucocitos Mononucleares , Mutación Puntual