Familial ApoB-specific familial hypobetalipoproteinemia in a patient with non-classical congenital adrenal hyperplasia. / Hipobetalipoproteinemia familiar ApoB específica en una paciente con hiperplasia suprarrenal congénita no clásica.

Familial hypobetalipoproteinaemia is a disorder of lipid metabolism characterized by low levels of total cholesterol, low-density lipoprotein cholesterol and apolipoprotein B. ApoB-related familial hypolipoproteinemia is an autosomal condition with a codominance inheritance pattern. Non-classical congenital adrenal hyperplasia is an autosomal recessive disorder due to mutations in the CYP21A2, a gene encoding for the enzyme 21-hydroxylase, which results in an androgen excess production from adrenal source. We here present the case of a 25-year-old woman with NCAH showing decreased levels of total-cholesterol, low-density lipoprotein cholesterol and triglycerides. Her parent had digestive symptoms and severe hepatic steatosis with elevated liver enzymes, as well as decreased levels of total and low-density lipoprotein cholesterol. A genetic-molecular study of the proband identified a mutation in the APOB gene, which allowed a diagnosis of heterozygous ApoB-related hypolipoproteinaemia to be made.

Molecular diagnosis of cystic fibrosis by RNA obtained from nasal epithelial cells.

BACKGROUND: The diagnosis of cystic fibrosis (CF) is established when characteristic clinical signs are coupled with biallelic CFTR pathogenic variants. No previously reported non-canonical splice site variants have to be considered as variants of uncertain significance unless their effect on splicing has been validated. METHODS: Two variants identified by next-generation sequencing were evaluated. We assayed their effects on splicing employing RNA analysis and real-time expression quantification from RNA obtained from the nasal epithelial cells of a patient with clinically suspected CF and of two patients with milder phenotypes (CFTR-related disorders). RESULTS: The variant c.164+2dup causes skipping of exon 2 (p.(Ser18_Glu54del)) and exon 2 plus 3 (p.(Ser18Argfs*16)) in CFTR mRNA. Exon 2 expression in the patient heterozygous for c.164+2dup was decreased to 7 % of the exon 2 expression in the controls. The synonymous variant c.1584G>A causes a partial skipping of exon 11. The exon 11 expression in the two patients heterozygous for this variant was 22 % and 42 % of that of the controls, respectively. CONCLUSION: We conclude that variant c.164+2dup affects mRNA processing and can be considered a CF-causing variant. The results of the functional assay also showed that the p.(Glu528=) variant, usually categorized as a neutral variant based on epidemiological data, partially affects mRNA processing in our patients. This finding would allow us to reclassify the variant as a CFTR-related variant with incomplete penetrance. RNA obtained from nasal epithelial cells is an easy and accurate tool for CFTR functional studies in patients with unclassified splice variants.

Whole-Exome Sequencing of 21 Families: Candidate Genes for Early-Onset High Myopia.

High myopia is the most severe and pathological form of myopia. It occurs when the spherical refractive error exceeds -6.00 spherical diopters (SDs) or the axial length (AL) of the eye is greater than 26 mm. This article focuses on early-onset high myopia, an increasingly common condition that affects children under 10 years of age and can lead to other serious ocular pathologies. Through the genetic analysis of 21 families with early-onset high myopia, this study seeks to contribute to a better understanding of the role of genetics in this disease and to propose candidate genes. Whole-exome sequencing studies with a panel of genes known to be involved in the pathology were performed in families with inconclusive results: 3% of the variants found were classified as pathogenic, 6% were likely pathogenic and the remaining 91% were variants of uncertain significance. Most of the families in this study were found to have alterations in several of the proposed genes. This suggests a polygenic inheritance of the pathology due to the cumulative effect of the alterations. Further studies are needed to validate and confirm the role of these alterations in the development of early-onset high myopia and its polygenic inheritance.

Whole-Exome Sequencing of 24 Spanish Families: Candidate Genes for Non-Syndromic Pediatric Keratoconus.

Keratoconus is a corneal dystrophy that is one of the main causes of corneal transplantation and for which there is currently no effective treatment for all patients. The presentation of this disease in pediatric age is associated with rapid progression, a worse prognosis and, in 15-20% of cases, the need for corneal transplantation. It is a multifactorial disease with genetic variability, which makes its genetic study difficult. Discovering new therapeutic targets is necessary to improve the quality of life of patients. In this manuscript, we present the results of whole-exome sequencing (WES) of 24 pediatric families diagnosed at the University Hospital La Paz (HULP) in Madrid. The results show an oligogenic inheritance of the disease. Genes involved in the structure, function, cell adhesion, development and repair pathways of the cornea are proposed as candidate genes for the disease. Further studies are needed to confirm the involvement of the candidate genes described in this article in the development of pediatric keratoconus.

MiR-151a: a robust endogenous control for normalizing small extracellular vesicle cargo in human cancer.

Small extracellular vesicles (sEVs) in the blood of cancer patients contain higher amounts of tumor markers than those identified as free-circulating. miRNAs have significant biomedical relevance due to their high stability and feasible detection. However, there is no reliable endogenous control available to measure sEVs-miRNA content, impairing the acquisition of standardized consistent measurements in cancer liquid biopsy. In this study, we identified three miRNAs from a panel of nine potential normalizers that emerged from a comprehensive analysis comparing the sEV-miRNA profile of six lung and ovarian human cancer cell lines in the absence of or under different conditions. Their relevance as normalizers was tested in 26 additional human cancer cell lines from nine different tumor types undergoing chemotherapy or radiotherapy treatment. The validation cohorts were comprised of 242 prospective plasma and ascitic fluid samples from three different human tumor types. Variability and normalization properties were tested in comparison to miR-16, the most used control to normalize free-circulating miRNAs in plasma. Our results indicate that miR-151a is consistently represented in small extracellular vesicles with minimal variability compared to miR-16, providing a novel normalizer to measure small extracellular vesicle miRNA content that will benefit liquid biopsy in cancer patients.

Whole Exome Sequencing of 20 Spanish Families: Candidate Genes for Non-Syndromic Pediatric Cataracts.

Non-syndromic pediatric cataracts are defined as opacification of the crystalline lens that occurs during the first years of life without affecting other organs. Given that this disease is one of the most frequent causes of reversible blindness in childhood, the main objective of this study was to propose new responsible gene candidates that would allow a more targeted genetic approach and expand our genetic knowledge about the disease. We present a whole exome sequencing (WES) study of 20 Spanish families with non-syndromic pediatric cataracts and a previous negative result on an ophthalmology next-generation sequencing panel. After ophthalmological evaluation and collection of peripheral blood samples from these families, WES was performed. We were able to reach a genetic diagnosis in 10% of the families analyzed and found genes that could cause pediatric cataracts in 35% of the cohort. Of the variants found, 18.2% were classified as pathogenic, 9% as likely pathogenic, and 72.8% as variants of uncertain significance. However, we did not find conclusive results in 55% of the families studied, which suggests further studies are needed. The results of this WES study allow us to propose LONP1, ACACA, TRPM1, CLIC5, HSPE1, ODF1, PIKFYVE, and CHMP4A as potential candidates to further investigate for their role in pediatric cataracts, and AQP5 and locus 2q37 as causal genes.

Serie de casos sobre la mortalidad a 30 días tras trombectomía o fibrinólisis. El reto de la dispersión demográfica / Case series on 30-day mortality after thrombectomy or fibrinolysis. The challenge of demographic dispersion

Objetivo: el presente estudio tiene como propósito conocer la supervivencia de la trombectomía o fibrinólisis de pacientes con criterios de código ictus al cabo de 30 días. Además, se propone observar si la dispersión demográfica por ubicación del hospital tiene alguna influencia en dicha relación para el paciente. Método: estudio longitudinal, retrospectivo y observacional. Pacientes dentro del código ictus en la provincia de Ourense (Galicia). Los participantes son 153 pacientes con una media de edad de 76 ± 12 años, de los cuales 83 son mujeres, reclutados durante 17 meses. Las variables a considerar incluyeron la arteria afectada, intervención fibrinólisis/trombectomía, mortalidad al cabo de 30 días, aplicación de angiotomografía computarizada (angio-TC), cambios en la National Institutes Health Stroke Scale (NIHSS) y tiempos en el servicio de urgencias en tres hospitales de la provincia (Ourense, O Barco y Verín). Resultados: el 12,9% de los pacientes fallecieron en los 30 días siguientes a partir del evento. El 54,2% (84 pacientes) fueron sometidos a fibrinólisis; el 19,4%, a trombectomía, y el 26,0% (40 pacientes) fueron tratados mediante intervención combinada (fibrinólisis + trombectomía). El 13,1% de los pacientes bajo fibrinólisis alcanzaron el exitus letalis al cabo de 30 días. El análisis de regresión logística determinó que hacer una trombectomía disminuye la probabilidad de fallecimiento, pero no de forma significativa. Conclusiones: no se encontró una relación significativa entre la aplicación de un tratamiento concreto (trombectomía/fibrinólisis) o su combinación y la tasa de mortalidad de los pacientes al cabo de 30 días.(AU)

Purpose: this study aims to analyze the survival and mortality rate after 30 days in patients with stroke code and fibrinolysis/thrombectomy. In addition, it is proposed to observe whether the demographic dispersion by the hospital’s location has any impact on this relationship for the patient. Method: longitudinal, retrospective and observational study. Patients with stroke code in the province of Ourense, Galicia. Subjects are 153 patients with a mean age of 76±12 years (83 women), recruited over 17 months. The variables to consider included the affected artery, fibrinolysis/thrombectomy intervention, mortality after 30 days, application of computerized tomography angiography, changes in the NIHSS (National Institutes Stroke Scale) and times in the casualty department in three hospitals in the province (Ourense, O Barco and Verín). Results: 12.9% of patients died within 30 days of the event. At least 54.2% of patients (84) received fibrinolysis, 19.4% underwent thrombectomy and at least 40 patients (26.0%) underwent a combined intervention (fibrinolysis + thrombectomy). A total of 13.1% of patients with fibrinolysis reached exitus letalis after 30 days. Logistic analysis determined that performing a thrombectomy reduces the probability of death, but not significantly. Conclusions: no statistically significant relationship was detected between the application of a specific treatment (thrombectomy/fibrinolysis) or its combination with the mortality rate of patients after 30 days. Ourense hospital that is closest to the thrombectomy referral centres shows P<0.005 in regard to mortality.(AU)

Identification and Functional Analysis of APOB Variants in a Cohort of Hypercholesterolemic Patients.

Mutations in APOB are the second most frequent cause of familial hypercholesterolemia (FH). APOB is highly polymorphic, and many variants are benign or of uncertain significance, so functional analysis is necessary to ascertain their pathogenicity. Our aim was to identify and characterize APOB variants in patients with hypercholesterolemia. Index patients (n = 825) with clinically suspected FH were analyzed using next-generation sequencing. In total, 40% of the patients presented a variant in LDLR, APOB, PCSK9 or LDLRAP1, with 12% of the variants in APOB. These variants showed frequencies in the general population lower than 0.5% and were classified as damaging and/or probably damaging by 3 or more predictors of pathogenicity. The variants c.10030A>G;p.(Lys3344Glu) and c.11401T>A;p.(Ser3801Thr) were characterized. The p.(Lys3344Glu) variant co-segregated with high low-density lipoprotein (LDL)-cholesterol in 2 families studied. LDL isolated from apoB p.(Lys3344Glu) heterozygous patients showed reduced ability to compete with fluorescently-labelled LDL for cellular binding and uptake compared with control LDL and was markedly deficient in supporting U937 cell proliferation. LDL that was carrying apoB p.(Ser3801Thr) was not defective in competing with control LDL for cellular binding and uptake. We conclude that the apoB p.(Lys3344Glu) variant is defective in the interaction with the LDL receptor and is causative of FH, whereas the apoB p.(Ser3801Thr) variant is benign.

[Analysis of cardiovascular risk factors in the stroke code. An approach in advanced ages.] / Análisis de los factores de riesgo cardiovascular en el código ictus. Una aproximación en edades avanzadas.

OBJECTIVE: The purpose of this study was to analyze the effect of cardiovascular risk factors (CVRF) in 153 patients who had suffered a stroke in the province of Ourense and where the stroke code had been activated.Its realization is part of the purpose of the authors to influence chronic pathology to prevent this specific event. METHODS: A longitudinal, retrospective and observational study was applied to 153 patients with a mean age of 76±12 years who had presented some type of stroke. The independent variables were classified as quantitative (international normalized ratio [INR], blood pressure and glycemia) and in qualitative (atrial fibrillation [AF], consumption of anticoagulants [ACO], smoking and blood lipid levels). The dependent variables were the type of stroke, the affected artery, and patient mortality after thirty days, six months, and one year. For the qualitative variables, the non-parametric verification method of Kruskal-Wallis (K-W) and Mann-Whitney (M-W) was used for comparison of means and for Chi-square association. RESULTS: INR was associated with the type of event and mortality at six and twelve months (p<0,001). Glycemia was related in a statistically significant way both with the type of event, the affected artery and with mortality in the three periods. Both AF and OACs were associated with the type of event and the artery affected, the latter also with one-year mortality. Age was related to mortality in the three periods without becoming significant at thirty days. Being a man could be a risk factor for thirty -day mortality (OR>1) and being a woman for one-year mortality. Regarding the type of intervention performed, undergoing fibrinolysis or thrombectomy increased the risk of mortality compared to combined treatment, the relationship between thrombectomy and increased mortality being statistically significant exclusively in the six-month period. CONCLUSIONS: According to the results obtained, the prevention of cerebrovascular events and secondary mortality should focus mainly on high blood glucose levels, the consumption of anticoagulants, INR, and the presence of AF as cardiovascular risk factors. Studies with a larger sample size are needed to establish if there really is an impact on mortality based on sex, as well as to determine with greater certainty if habits such as smoking, poor diet and other factors play a relevant role.

OBJETIVO: El presente estudio tuvo como propósito analizar el efecto de los factores de riesgo cardiovascular (FRCV) en 153 pacientes de la provincia de Ourense que habían sufrido ictus, donde se había activado el código ictus. Su realización parte del propósito de los autores de incidir en la patología crónica para prevenir este evento en concreto. METODOS: Se realizó un estudio longitudinal, retrospectivo y observacional aplicado a 153 pacientes con una media de edad de 76±12 años que habían presentado algún de tipo de ictus. Las variables independientes fueron clasificadas en cuantitativas (índice internacional normalizado [INR], presión arterial y glucemia) y en cualitativas (fibrilación auricular [FA], consumo de anticoagulantes [ACO], hábito de fumar y nivel de lípidos en sangre). Las variables dependientes fueron el tipo de ictus, la arteria afectada y la mortalidad de paciente al cabo de treinta días, seis meses y un año. Para las variables cualitativas se utilizó el método de comprobación no paramétrico de Kruskal-Wallis (K-W) y Mann-Whitney (M-W) para comparación de medias y para asociación Chi-cuadrado. RESULTADOS: El INR resultó asociado con el tipo de evento así como con la mortalidad a seis y doce meses (p<0,001). La glucemia se relacionó de forma estadísticamente significativa tanto con el tipo de evento como con la arteria afectada, así como con la mortalidad en los tres periodos. Tanto la FA como los ACO se asociaron con el tipo de evento y con la arteria afecta, los últimos además con la mortalidad a un año. La edad se relacionó con la mortalidad en los tres periodos sin llegar a ser significativa a los treinta días. Ser hombre podría ser un factor de riesgo (OR>1) para la mortalidad a treinta días y ser mujer para la mortalidad a un año . En cuanto al tipo de intervención realizada someterse a fibrinolisis o a trombectomía aumentó el riesgo de mortalidad frente al tratamiento combinado, siendo estadísticamente significativa la relación entre trombectomía y aumento de mortalidad exclusivamente en el periodo de seis meses. CONCLUSIONES: Según los resultados obtenidos la prevención de eventos cerebrovasculares y mortalidad secundaria a estos se debe centrar principalmente en los niveles elevados de glucemia, el consumo de anticoagulantes, INR y presencia de FA como factores de riesgo cardiovascular. Se precisan estudios de mayor tamaño muestral para establecer si realmente existe un impacto en la mortalidad en función del sexo así como determinar con mayor certeza si hábitos como el fumar, la mala alimentación y otros factores desempeñan un papel relevante.

Análisis de los factores de riesgo cardiovascular en el código ictus. Una aproximación en edades avanzadas / Analysis of cardiovascular risk factors in the stroke code. An approach in advanced ages

FUNDAMENTOS: El presente estudio tuvo como propósito analizar el efecto de los factores de riesgo cardiovascular (FRCV) en 153 pacientes de la provincia de Ourense que habían sufrido ictus, donde se había activado el código ictus. Su realización parte del propósito de los autores de incidir en la patología crónica para prevenir este evento en concreto. MÉTODOS: Se realizó un estudio longitudinal, retrospectivo y observacional aplicado a 153 pacientes con una media de edad de 76±12 años que habían presentado algún de tipo de ictus. Las variables independientes fueron clasificadas en cuantitativas (índice internacional normalizado [INR], presión arterial y glucemia) y en cualitativas (fibrilación auricular [FA], consumo de anticoagulantes [ACO], hábito de fumar y nivel de lípidos en sangre). Las variables dependientes fueron el tipo de ictus, la arteria afectada y la mortalidad de paciente al cabo de treinta días, seis meses y un año. Para las variables cualitativas se utilizó el método de comprobación no paramétrico de Kruskal-Wallis (K-W) y Mann-Whitney (M-W) para comparación de medias y para asociación Chi-cuadrado. RESULTADOS: El INR resultó asociado con el tipo de evento así como con la mortalidad a seis y doce meses (p<0,001). La glucemia se relacionó de forma estadísticamente significativa tanto con el tipo de evento como con la arteria afectada, así como con la mortalidad en los tres periodos. Tanto la FA como los ACO se asociaron con el tipo de evento y con la arteria afecta, los últimos además con la mortalidad a un año. La edad se relacionó con la mortalidad en los tres periodos sin llegar a ser significativa a los treinta días. Ser hombre podría ser un factor de riesgo (OR>1) para la mortalidad a treinta días y ser mujer para la mortalidad a un año . En cuanto al tipo de intervención realizada someterse a fibrinolisis o a trombectomía aumentó el riesgo de mortalidad frente al tratamiento combinado, siendo estadísticamente...(AU)

BACKGROUND: The purpose of this study was to analyze the effect of cardiovascular risk factors (CVRF) in 153 patients who had suffered a stroke in the province of Ourense and where the stroke code had been activated.Its realization is part of the purpose of the authors to influence chronic pathology to prevent this specific event. METHODS: A longitudinal, retrospective and observational study was applied to 153 patients with a mean age of 76±12 years who had presented some type of stroke. The independent variables were classified as quantitative (international normalized ratio [INR], blood pressure and glycemia) and in qualitative (atrial fibrillation [AF], consumption of anticoagulants [ACO], smoking and blood lipid levels). The dependent variables were the type of stroke, the affected artery, and patient mortality after thirty days, six months, and one year. For the qualitative variables, the non-parametric verification method of Kruskal-Wallis (K-W) and Mann-Whitney (M-W) was used for comparison of means and for Chi-square association. RESULTS: INR was associated with the type of event and mortality at six and twelve months (p<0,001). Glycemia was related in a statistically significant way both with the type of event, the affected artery and with mortality in the three periods. Both AF and OACs were associated with the type of event and the artery affected, the latter also with one-year mortality. Age was related to mortality in the three periods without becoming significant at thirty days. Being a man could be a risk factor for thirty -day mortality (OR>1) and being a woman for one-year mortality. Regarding the type of intervention performed, undergoing fibrinolysis or thrombectomy increased the risk of mortality compared to combined treatment, the relationship between thrombectomy and increased mortality being statistically significant exclusively in the six-month period...(AU)

Impact of COVID-19 on University Students: An Analysis of Its Influence on Psychological and Academic Factors.

The irruption of COVID-19 has had different consequences on mental health in the youth population. Specifically, the sector made up of university students has suffered an abrupt change of teaching modality because of the pandemic. As such, this paper aims to analyze the impact that COVID-19 has had on different personal factors of students: (i) satisfaction with life; (ii) lived uncertainty; (iii) depression, anxiety, and stress, as well as factors related to academic development; (iv) motivation and the creation of teaching and learning strategies during this period; and (v) the perception of the degree of adaptability to the new scenario brought about by the university system. For this purpose, a cross-sectional quantitative design was advocated through the elaboration of an SEM model, which included 1873 university students from Andalusian Universities (Spain). The results reflected the strong negative impact that the pandemic had, especially on the levels of life satisfaction and the indices of depression, anxiety, and stress of the students. Likewise, the findings reflected the relevance of the correct adaptability on the part of the university to these new circumstances. It is necessary for university institutions to focus their efforts on quality attention to students, in order to establish fluid communication with them and to adapt to their academic and personal needs.

Broadening the phenotypic spectrum of EVEN-PLUS syndrome through identification of HSPA9 pathogenic variants in the original EVE dysplasia family and two sibs with milder facial phenotype.

EVEN-PLUS syndrome is a rare autosomal recessive disorder caused by biallelic pathogenic variants in the mitochondrial chaperone called mortalin, encoded by HSPA9. This genetic disorder, presenting with several overlapping features with CODAS syndrome, is characterized by the involvement of the Epiphyses, Vertebrae, Ears, and Nose (EVEN), PLUS associated findings. Only five individuals presenting with the EVEN-PLUS phenotype and biallelic variants in HSPA9 have been published. Here, we expand the phenotypic and molecular spectrum associated with this disorder, reporting two sibs with a milder phenotype and compound heterozygous pathogenic variants (a recurrent variant and a novel one). Also, we confirm a homozygous pathogenic variant in the family originally reported as EVE dysplasia.

Hereditary spastic paraplegia associated with a novel homozygous intronic noncanonical splice site variant in the AP4B1 gene.

Pathogenic variants in the AP4B1 gene lead to a rare form of hereditary spastic paraplegia (HSP) known as SPG47. We report on a patient with a clinical suspicion of complicated HSP of the lower limbs with intellectual disability, as well as a novel homozygous noncanonical splice site variant in the AP4B1 gene, in which the effect on splicing was validated by RNA analysis. We sequenced 152 genes associated with HSP using Next-Generation Sequencing (NGS). We isolated total RNA from peripheral blood and generated cDNA using reverse transcription-polymerase chain reaction (RT-PCR). A region of AP4B1 mRNA was amplified by PCR and the fragments obtained were purified from the agarose gel and sequenced. We found a homozygous variant of uncertain significance in the AP4B1 gene NM_006594.4: c.1511-6C>G in the proband. Two different AP4B1 mRNA fragments were obtained in the patient and his carrier parents. The shorter fragment was the predominant fragment in the patient and revealed a deletion with skipping of the AP4B1 exon 10. The patient's longer fragment corresponded to an insertion of the last five nucleotides of AP4B1 intron 9. We confirmed that this variant affects the normal splicing of RNA, sustaining the molecular diagnosis of SPG47 in the patient.

Novel SLC12A2-ROS1 Fusion in Non-Small Cell Lung Cancer with a Significant Response to Crizotinib: The Importance of Choosing the Appropriate Next-Generation Sequencing Assay.

Identifying the druggable target is crucial for patients with nonsquamous advanced non-small cell lung cancer (NSCLC). This article adds to the spectrum of ROS1 fusion cases described in NSCLC. We describe a novel SLC12A2-ROS1 rearrangement that has not been previously reported in other cancers: a fusion that has clinical and radiological sensitivity to crizotinib. Fluorescence in situ hybridization detected the SLC12A2-ROS1 fusion and it was confirmed through hybrid capture-based next-generation sequencing (NGS); however, the fusion could not be detected by amplicon-based assay. The success of implementing NGS into routine clinical practice depends on the accuracy of testing. The test's methodological features should then be considered because they significantly affect the results. Given this patient's response to crizotinib, identifying patients with undescribed ROS1 fusions has important therapeutic implications. KEY POINTS: This is the first known description of an SLC12A2-ROS1 fusion. Considering the patient's clinical features and tumor response observed after crizotinib therapy, the authors confirm that this new rearrangement has relevant clinical impact for patients with non-small cell lung cancer. The success of implementing next-generation sequencing (NGS) into routine clinical practice depends on the accuracy of the testing. Different assays and NGS platforms can achieve differing results. Each assay's limitations need to be considered to ensure the quality of precision medicine in clinical practice.

Delineation of the clinical and radiological features of Stuve-Wiedemann syndrome childhood survivors, four new cases and review of the literature.

Stuve-Wiedemann syndrome (SWS; MIM 601559) is a rare autosomal recessive disease caused by mutations in the leukemia inhibitor factor receptor gene (LIFR). Common clinical and radiological findings are often observed, and high neonatal mortality occurs due to respiratory distress and hyperthermic episodes. Despite initially considered as a lethal disorder during the newborn period, in recent years, several SWS childhood survivors have been reported. We report a detailed clinical and radiological characterization of four unrelated childhood SWS molecularly confirmed patients and review 22 previously reported childhood surviving cases. We contribute to the definition of the childhood survival phenotype of SWS, emphasizing the evolving phenotype, characterized by skeletal abnormalities with typical radiological findings, distinctive dysmorphic features, and dysautonomia. Based on the typical features and clinical course, early diagnosis is possible and crucial to plan appropriate management and prevent potential complications. Genetic confirmation is advisable in order to improve genetic counseling to the patients and their families.

Familial hypercholesterolemia: A single-nucleotide variant (SNV) in mosaic at the low density lipoprotein receptor (LDLR).

BACKGROUND AND AIMS: Familial hypercholesterolemia is most frequently caused by genetic variants in the LDLR gene. Most of LDLR pathogenic variants are missense, followed by splicing and deletion/insertions variants. Mosaicism is a genetic condition in which an individual shows more than one clone of cells with different genotypes. The objective of this article was the molecular characterization of a patient with hypercholesterolemia. METHODS AND RESULTS: Genetic analysis of DNA from peripheral blood and saliva was performed by NGS, Sanger sequencing and pyrosequencing technologies. NGS analysis detected the pathogenic variant LDLR:c.1951G > T:p.(Asp651Tyr) in 9%-12% of reads. The presence of the variant was confirmed by pyrosequencing analysis. The variant found was functional characterized using an in vitro model (CHO-ldlA7 cells). Activity and expression of cell surface LDLR were measured by flow cytometry. Colocalization LDLR-Dil-LDL was detected by immunofluorescence. The LDLR activity showed 80% uptake, 50% binding and 53% expression of cell surface LDLR regarding wild type. CONCLUSIONS: Herein, we report the first case of a mosaic single nucleotide variant affecting the LDLR gene in a patient with familial hypercholesterolemia. As it has been described for other pathologies, mosaicism could be underestimated in FH and its detection will improve with the introduction of NGS technologies in the diagnostic routine.

Loss-of-function mutation of PCSK9 as a protective factor in the clinical expression of familial hypercholesterolemia: A case report.

RATIONALE: Proprotein convertase subtilisin/kexin 9 or PCSK9 is a protein whose main function is to regulate the number of low-density lipoprotein receptors (LDLR) present on the cell surface. Loss-of-function mutations in PCSK9 have been related to low LDL-cholesterol levels and a decrease in the risk of cardiovascular events. PATIENT CONCERNS: We present the case of a 27-year-old woman, offspring of a patient with familial homozygous hypercholesterolemia, who presented with mild-moderate hypercholesterolemia. DIAGNOSIS: Genetic analysis was performed by next generation sequencing using a customized panel of 198 genes. Sanger sequencing was used to confirm the presence of the variants of interest. The genetic analysis showed a pathogenic heterozygous mutation in LDLR [exon 6:c.902A>G:p(Asp301Gly)], as well as a loss-of-function heterozygous variant in PCSK9 [exon1:c.137 G>T:p.(Arg46Leu)]. The genetic analysis of the index case's mother revealed compound heterozygosity for 2 different mutations in LDLR [c.902A>G:p.(Asp301Gly); c.1646G>T:p.(Gly549Val)] in exon 6 and in exon 11, respectively, and the same loss-of-function variant in PCSK9 that had been found in her daughter [(PCSK9:exon1:c.137G>T:p.(Arg46Leu)]. The maternal grandfather of the index case presented the same genetic variants as his granddaughter. INTERVENTIONS: The index case did not receive any specific treatment for hypercholesterolemia. The loss-of-function variant in PCSK9 protected her from higher LDL-cholesterol levels, provided she kept partial activity of the LDLR. In her mother, instead, a PCSK9 inhibitor was tried but failed to achieve lipid control. The reason for this may be the complete absence in LDL receptor activity. LDL apheresis was started afterwards, resulting in adequate lipid level control. OUTCOMES: To the date, the index case has achieved to maintain adequate total and LDL-cholesterol levels without any other intervention. She has had no known cardiovascular complication. LESSONS: Loss-of-function mutations in PCSK9 could protect from developing more severe forms of hypercholesterolemia. The finding of these mutations (LDLR-PCSK9) in three consecutive generations could imply an adaptive mechanism against the development of hypercholesterolemia.

Effect of the Flipped Classroom and Gamification Methods in the Development of a Didactic Unit on Healthy Habits and Diet in Primary Education.

Currently, there are several methodological models that have broken into different disciplines of knowledge with the aim of making the teaching/learning process more dynamic, active and participatory for students. This is the case of Flipped Classroom, which is based on a mixed approach between e-learning and face-to-face teaching, as well as gamification, which bases its didactic principles on the recreational components of the games. Within this context, the aim of this research is to observe what effect the application of Flipped Classroom and gamification has in the development of motivation, autonomy and self-regulation towards learning through a didactic unit on healthy habits and diet in 202 students of 6th grade of Primary School from four different schools (public and state-subsidized) in the city of Granada (Spain). For this purpose, a methodological design was used with pre-test and post-test to check the effects of the experience on the students. The findings obtained showed that the application of these methods promoted an increase in students' motivation, as well as in their autonomy and self-regulation when facing the contents of the subject. For this reason, it is advocated that there is a need to continue promoting a quality and innovative educational practice according to the figure of the student today.

Impact of Problematic Smartphone Use and Instagram Use Intensity on Self-Esteem with University Students from Physical Education.

Mobile devices are a revolutionary element that offer many possibilities, although they can also cause problems for users. This is the case with the development of addictive behaviors that can affect personal well-being. The purpose of this paper has been to analyze the influence of smartphone addiction and Instagram use intensity on the self-esteem of Physical Education students. A cross-sectional research design was adopted by applying an online survey to a sample of undergraduate students (n = 385). The results showed that gender and age were factors that influenced the problematic use of the smartphone. In turn, there was a significant positive correlation between smartphone addiction and Instagram use intensity. The influence of smartphone addiction on students' self-esteem was also highlighted. In contrast, Instagram use intensity did not affect self-esteem. Finally, the findings are discussed, and the main implications of the study are established, where physical education students take on a special role in order to avoid the improper use of smartphones and Instagram through sport.

Use of Instagram by Pre-Service Teacher Education: Smartphone Habits and Dependency Factors.

It is increasingly common to upload photographs on the Internet of sports practices carried out. However, this behavior can be related to smartphone addiction, which has become a global problem. In turn, the intensive use of the Instagram social network has begun to be linked to addictive behaviors on mobile devices. The purposes of this paper were to analyze the Instagram usage habits of future primary school teachers, to determine the influence of sociodemographic factors on intensive Instagram use and smartphone addiction, and to determine the influence of intensive Instagram use on smartphone addiction. For this purpose, a transversal design was adopted where two standardized scales were applied to a sample of university students of the Primary Education Degree of the University of Granada, Spain (n = 385). The results showed that the type of image most uploaded to this social network was the selfie, well above sport. Furthermore, the structural equation model highlighted the significant influence of the intensive use of Instagram and smartphone addiction. Finally, the implications and findings of this study are discussed, highlighting the importance of generating healthy habits regarding the use of technology.