MicroRNA biogenesis is broadly disrupted by inhibition of the splicing factor SF3B1.

In animals, microRNA (miRNA) biogenesis begins with cotranscriptional cleavage of the primary (pri-)miRNA by the Microprocessor complex. Cotranscriptional splicing has been shown to influence Microprocessor cleavage when miRNAs are hosted in introns of protein-coding pri-miRNAs, but the impact of splicing on production of miRNAs hosted in long non-coding (lnc)RNAs is largely unknown. Here, we investigated the role of splicing in the biogenesis of miR-122, an lncRNA-hosted, highly expressed, medically important, liver-specific miRNA. We found that splicing inhibition by the SF3B1 inhibitor pladienolide B (PlaB) led to strong and rapid reduction in transcription of endogenous, but not plasmid-encoded, pri-miR-122, resulting in reduced production of mature miR-122. To allow detection of rapid changes in miRNA biogenesis despite the high stability of mature miRNAs, we used SLAMseq to globally quantify the effects of short-term splicing inhibition on miRNA synthesis. We observed an overall decrease in biogenesis of mature miRNAs following PlaB treatment. Surprisingly, miRNAs hosted in exons and introns were similarly affected. Together, this study provides new insights into the emerging role of splicing in transcription, demonstrating novel biological importance in promotion of miR-122 biogenesis from an lncRNA, and shows that SF3B1 is important for global miRNA biogenesis.

Highly Sensitive Microsatellite Instability and Immunohistochemistry Assessment in Endometrial Aspirates as a Tool for Cancer Risk Individualization in Lynch Syndrome.

Women with Lynch syndrome (LS) are at increased risk of endometrial cancer (EC), among other tumors, and are characterized by mismatch repair (MMR) deficiency and microsatellite instability (MSI). While risk-reducing gynecologic surgeries effectively decrease EC incidence, doubts arise regarding the appropriate timing of the surgery. We explored the usefulness of highly sensitive MSI (hs-MSI) assessment in endometrial aspirates for individualizing gynecologic surveillance in LS carriers. Ninety-three women with LS, 25 sporadic EC patients (9 MMR-proficient and 16 MMR-deficient), and 30 women with benign gynecologic disease were included in this study. hs-MSI was assessed in prospectively collected endometrial aspirates in 67 LS carriers, EC cases, and controls. MMR, PTEN, ARID1A, and PAX2 protein expression patterns were evaluated in the LS samples. Follow-up aspirates from 8 LS carriers were also analyzed. Elevated hs-MSI scores were detected in all aspirates from MMR-deficient EC cases (3 LS and 16 sporadic) and negative in aspirates from controls and MMR-proficient EC cases. Positive hs-MSI scores were also detected in all 4 LS aspirates reported as complex hyperplasia. High hs-MSI was also present in 10 of 49 aspirates (20%) from LS carriers presenting a morphologically normal endometrium, where MMR protein expression loss was detected in 69% of the samples. Interestingly, the hs-MSI score was positively correlated with MMR-deficient gland density and the presence of MMR-deficient clusters, colocalizing PTEN and ARID1A expression loss. High hs-MSI scores and clonality were evidenced in 2 samples collected up to 4 months before EC diagnosis; hs-MSI scores increased over time in 5 LS carriers, whereas they decreased in a patient with endometrial hyperplasia after progestin therapy. In LS carriers, elevated hs-MSI scores were detected in aspirates from premalignant and malignant lesions and normal endometrium, correlating with MMR protein loss. hs-MSI assessment and MMR immunohistochemistry may help individualize EC risk assessment in women with LS.

Role of psychological background in cancer susceptibility genetic testing distress: It is not only about a positive result.

Clinical and familial factors predict psychological distress after genetic testing for cancer susceptibility. However, the contribution of an individual's psychological background to such distress is unclear. This study aims to analyze the psychological impact of genetic testing and to identify the profile of individuals at higher risk. This is a longitudinal multicenter study of individuals undergoing genetic testing for cancer susceptibility. Demographic, clinical, genetic, familial, and psychological (personality types, cancer worry) characteristics were assessed by validated questionnaires the day of genetic testing. Distress, uncertainty, and positive experience perception (MICRA scale) were evaluated at the results disclosure visit, and 3 and 12 months afterwards. Multivariate analysis was performed. A total of 714 individuals were included. A high neuroticism score, high baseline cancer worry, and a positive genetic test result were independently associated with higher psychological impact (p-value < 0.05). The highest risk profile (10% of the cohort) included women with high level of neuroticism and a positive result. Uncertainty was mainly associated with a high level of neuroticism, regardless of the genetic test result. A holistic approach to personalized germline genetic counseling should include the assessment of personality dimensions.

Patients' and professionals' perspective of non-in-person visits in hereditary cancer: predictors and impact of the COVID-19 pandemic.

PURPOSE: To identify predictors of patient acceptance of non-in-person cancer genetic visits before and after the COVID-19 pandemic and assess the preferences of health-care professionals. METHODS: Prospective multicenter cohort study (N = 578, 1 February 2018-20 April 2019) and recontacted during the COVID-19 lockdown in April 2020. Health-care professionals participated in May 2020. Association of personality traits and clinical factors with acceptance was assessed with multivariate analysis. RESULTS: Before COVID-19, videoconference was more accepted than telephone-based visits (28% vs. 16% pretest, 30% vs. 19% post-test). Predictors for telephone visits were age (pretest, odds ratio [OR] 10-year increment = 0.79; post-test OR 10Y = 0.78); disclosure of panel testing (OR = 0.60), positive results (OR = 0.52), low conscientiousness group (OR = 2.87), and post-test level of uncertainty (OR = 0.93). Predictors for videoconference were age (pretest, OR 10Y = 0.73; post-test, OR 10Y = 0.75), educational level (pretest: OR = 1.61), low neuroticism (pretest, OR = 1.72), and post-test level of uncertainty (OR = 0.96). Patients' reported acceptance for non-in-person visits after COVID-19 increased to 92% for the pretest and 85% for the post-test. Health-care professionals only preferred non-in-person visits for disclosure of negative results (83%). CONCLUSION: These new delivery models need to recognize challenges associated with age and the psychological characteristics of the population and embrace health-care professionals' preferences.

High-sensitivity microsatellite instability assessment for the detection of mismatch repair defects in normal tissue of biallelic germline mismatch repair mutation carriers.

INTRODUCTION: Lynch syndrome (LS) and constitutional mismatch repair deficiency (CMMRD) are hereditary cancer syndromes associated with mismatch repair (MMR) deficiency. Tumours show microsatellite instability (MSI), also reported at low levels in non-neoplastic tissues. Our aim was to evaluate the performance of high-sensitivity MSI (hs-MSI) assessment for the identification of LS and CMMRD in non-neoplastic tissues. MATERIALS AND METHODS: Blood DNA samples from 131 individuals were grouped into three cohorts: baseline (22 controls), training (11 CMMRD, 48 LS and 15 controls) and validation (18 CMMRD and 18 controls). Custom next generation sequencing panel and bioinformatics pipeline were used to detect insertions and deletions in microsatellite markers. An hs-MSI score was calculated representing the percentage of unstable markers. RESULTS: The hs-MSI score was significantly higher in CMMRD blood samples when compared with controls in the training cohort (p<0.001). This finding was confirmed in the validation set, reaching 100% specificity and sensitivity. Higher hs-MSI scores were detected in biallelic MSH2 carriers (n=5) compared with MSH6 carriers (n=15). The hs-MSI analysis did not detect a difference between LS and control blood samples (p=0.564). CONCLUSIONS: The hs-MSI approach is a valuable tool for CMMRD diagnosis, especially in suspected patients harbouring MMR variants of unknown significance or non-detected biallelic germline mutations.

Alcohol use and risk factors for self-harm behavior in Spanish adolescents. / Consumo de alcohol y factores de riesgo de conductas autolesivas en adolescentes españoles.

Self-harm behaviors in children and adolescents constitute an important public health problem with prevalence figures in the clinical population between 40 and 80%. The objectives of the study were to analyze and compare the Spanish sub-samples of two studies, SEYLE and WE-STAY to determine prevalence, self-harm patterns and factors associated with self-harm behaviors, notably the use of alcohol or drugs. The questionnaires used in both studies were the Global School Health Survey (GSHS), the Beck Depression Inventory (BDI-II), the Strengths and Difficulties Questionnaire (SDQ). The self-harm behaviors were evaluated with a modified 6-item version of s the Deliberate Self-Harm Inventory (DSHI). The independence of the study's categorical variables was assessed using the Chi-square test. The change in the relative risk of self-harm between the SEYLE study and WE-STAY was evaluated through the odds ratio (OR) calculation. Two different logistic regression models were calculated in order to establish the factors associated with self-harm behaviors in each study. In the present study, the rates of DSH vary according to study and sex, ranging from 0.58% to 2.08%, and different patterns of self-harm are evidenced by sex, with males self-injuring more frequently by self-inflicted blows and burns, while young women more often cut themselves. The presence of depressive symptoms and alcohol use were the factors most strongly associated with an increased risk of DSH.

Las conductas autolesivas en niños y adolescentes constituyen un importante problema de salud pública con cifras de prevalencia en la población clínica entre el 40 y 80%. Los objetivos del estudio son analizar y comparar las submuestras españolas de dos trabajos, SEYLE y WE-STAY, para conocer la prevalencia, los patrones de autolesión y los factores asociados a las conductas autolesivas, en particular el consumo de alcohol o drogas. Los cuestionarios utilizados en ambos estudios fueron la Encuesta Global de Salud Escolar (GSHS), el Inventario de Depresión de Beck (BDI-II), el Cuestionario de Fortalezas y Dificultades (SDQ). Los comportamientos autolesivos fueron evaluados con una versión modificada de 6 ítems basada en el Inventario de Autolesiones Deliberadas (DSHI). La independencia de las variables categóricas del estudio se evaluó mediante la prueba Ji-Cuadrado. El cambio en el riesgo relativo de autolesión entre el estudio SEYLE y WE-STAY, se evaluó a través del cálculo de odds ratio (OR). Se calcularon dos modelos de regresión logística diferentes con el fin de establecer los factores asociados con comportamientos autolesivos en cada estudio. En el presente estudio las tasas de DSH varían en función del estudio y del sexo en un rango entre 0,58% y 2,08%, presentando patrones de autolesiones diferentes según el sexo, los hombres se autolesionaron más frecuentemente mediante golpes autoinfligidos y quemaduras, mientras que las mujeres se hicieron más frecuentemente cortes. La presencia de síntomas depresivos y el consumo de alcohol fueron los factores asociados de forma más robusta a un mayor riesgo de DSH.

Comprehensive analysis and ACMG-based classification of CHEK2 variants in hereditary cancer patients.

CHEK2 variants are associated with intermediate breast cancer risk, among other cancers. We aimed to comprehensively describe CHEK2 variants in a Spanish hereditary cancer (HC) cohort and adjust the American College of Medical Genetics and Genomics and the Association for Molecular Pathology (ACMG-AMP) guidelines for their classification. First, three CHEK2 frequent variants were screened in a retrospective Hereditary Breast and Ovarian Cancer cohort of 516 patients. After, the whole CHEK2 coding region was analyzed by next-generation sequencing in 1848 prospective patients with HC suspicion. We refined ACMG-AMP criteria and applied different combined rules to classify CHEK2 variants and define risk alleles. We identified 10 CHEK2 null variants, 6 missense variants with discordant interpretation in ClinVar database, and 35 additional variants of unknown significance. Twelve variants were classified as (likely)-pathogenic; two can also be considered "established risk-alleles" and one as "likely risk-allele." The prevalence of (likely)-pathogenic variants in the HC cohort was 0.8% (1.3% in breast cancer patients and 1.0% in hereditary nonpolyposis colorectal cancer patients). Here, we provide ACMG adjustment guidelines to classify CHEK2 variants. We hope that this study would be useful for variant classification of other genes with low effect variants.

Early Psychological Correlates Associated With COVID-19 in A Spanish Older Adult Sample.

OBJECTIVE: Main aims of the study are to examine the early psychological correlates associated with the COVID-19 pandemic and lockdown on the mental health of a Spanish older adult sample and to analyze the influence of past mental disorder (PMD) and current mental disorder (CMD) on those correlates. METHODS: Cross-sectional study based on an online snowball recruiting questionnaire. Psychological correlates assessed with the Depression, Anxiety, and Stress Scale (DASS-21) and Impact of Event Scale (IES). Binary and multinomial logistic regression models were used to identify risk and protective factors. RESULTS: Final sample included 2,194 individuals aged 60 years or more (mean age [SD]: 65.62 [5.05]; females: 1,198 [54.6%]). There were 342 (15.6%) individuals who reported a PMD and 162 (7.4%) who reported a CMD. Avoidant (32.1%) and depressive (25.6%) styles were the most prevalent, regardless of mental health status. Main risk factors for negative affectivity were female gender and history CMD or PMD. However, job stability and the ability to enjoy free time were generally associated with better outcomes. No differences were found in psychological correlates between those with no lifetime history of mental disorder versus PMD on the DASS-21 or IES. However, CMD was associated with higher anxiety scores on the DASS-21 (odds ratio: 1.838, p < .001). CONCLUSION: Regardless of mental status, avoidant and depressive styles were the most prevalent in this older adult sample. Main protective factor in all subgroups was the ability to enjoy free time, whereas the main risk factors were being female and current or past history of mental disorder.

Does multilocus inherited neoplasia alleles syndrome have severe clinical expression?

IMPORTANCE: Genetic testing of hereditary cancer using comprehensive gene panels can identify patients with more than one pathogenic mutation in high and/or moderate-risk-associated cancer genes. This phenomenon is known as multilocus inherited neoplasia alleles syndrome (MINAS), which has been potentially linked to more severe clinical manifestations. OBJECTIVE: To determine the prevalence and clinical features of MINAS in a large cohort of adult patients with hereditary cancer homogeneously tested with the same gene panel. PATIENTS AND METHODS: A cohort of 1023 unrelated patients with suspicion of hereditary cancer was screened using a validated panel including up to 135 genes associated with hereditary cancer and phakomatoses. RESULTS: Thirteen (1.37%) patients harbouring two pathogenic mutations in dominant cancer-predisposing genes were identified, representing 5.7% (13/226) of patients with pathogenic mutations. Most (10/13) of these cases presented clinical manifestations associated with only one of the mutations identified. One case showed mutations in MEN1 and MLH1 and developed tumours associated with both cancer syndromes. Interestingly, three of the double mutants had a young age of onset or severe breast cancer phenotype and carried mutations in moderate to low-risk DNA damage repair-associated genes; two of them presented biallelic inactivation of CHEK2. We included these two patients for the sake of their clinical interest although we are aware that they do not exactly fulfil the definition of MINAS since both mutations are in the same gene. CONCLUSIONS AND RELEVANCE: Genetic analysis of a broad cancer gene panel identified the largest series of patients with MINAS described in a single study. Overall, our data do not support the existence of more severe manifestations in double mutants at the time of diagnosis although they do confirm previous evidence of severe phenotype in biallelic CHEK2 and other DNA repair cancer-predisposing genes.

Old drugs, new medication. / Viejas drogas, nuevos fármacos.

Editorial of vol 32(1).

Editorial del vol 32(1).

Novel POLE pathogenic germline variant in a family with multiple primary tumors results in distinct mutational signatures.

We describe a family in which four siblings exhibited multiple or classic colonic polyposis with or without colorectal carcinoma (CRC). One female developed three primary tumors, including CRC and carcinomas of the ovary and breast. Whole-exome sequencing of germline DNA from affected and unaffected individuals revealed a novel missense mutation in the exonuclease domain of POLE (c.833C>A; p.Thr278Lys) associated with a highly penetrant, autosomal-dominant inheritance pattern. Functional studies in yeast and demonstration of a high mutational burden in the available tumors confirmed the pathogenicity of the novel variant. Prominent POLE-deficient somatic mutational signatures were seen in the CRCs, but in contrast, a mutational signature typical of concomitant tumoral loss of POLE and mismatch-repair function (POLE-exo* /MSI) was noted in the breast cancer. The breast cancer also showed distinctive pathological characteristics that reflect the presence of both the germline POLE variant and the secondary somatic MMR alterations.

Lin28a uses distinct mechanisms of binding to RNA and affects miRNA levels positively and negatively.

Lin28a inhibits the biogenesis of let-7 miRNAs by triggering the polyuridylation and degradation of their precursors by terminal uridylyltransferases TUT4/7 and 3'-5' exoribonuclease Dis3l2, respectively. Previously, we showed that Lin28a also controls the production of neuro-specific miRNA-9 via a polyuridylation-independent mechanism. Here we reveal that the sequences and structural characteristics of pre-let-7 and pre-miRNA-9 are eliciting two distinct modes of binding to Lin28a. We present evidence that Dis3l2 controls miRNA-9 production. Finally, we show that the constitutive expression of untagged Lin28a during neuronal differentiation in vitro positively and negatively affects numerous other miRNAs. Our findings shed light on the role of Lin28a in differentiating cells and on the ways in which one RNA-binding protein can perform multiple roles in the regulation of RNA processing.

Detection of synthetic cannabinoid intoxication in the Emergency Department: when routine toxicological tests are not enough. / Detección de la intoxicación por cannabinoides sintéticos en Urgencias: cuando las pruebas toxicológicas rutinarias no bastan.

Letter to the editor.

Carta al Editor.

Predictive factors of alcohol consumption in adolescents: data from 1-year follow-up prospective study. / Factores predictores del consumo de alcohol en adolescentes: datos de un estudio prospectivo de 1 año de seguimiento.

Alcohol use/abuse is a health problem in adolescents. The last Survey on use of drugs in Secondary Schoolers carried out in Spain (ESTUDES 2014-2015), reveals that 76.8% of adolescents aged 14 to 18 years consumed alcohol in the previous year and 68.2% in the last month. The aim of this study is to determine the medium-term factors associated with alcohol consumption in a sample of Spanish adolescents. The present study was carried out as a part of the Saving and Empowering Young Lives project in Europe (SEYLE) project. The final sample was composed of 708 students, assessed at two times [basal (T0) and one year later (T1)] [males: 51.98%, basal mean age (SD)=4.43 (0.67)]. Univariate and multivariate regression analyses were performed in order to investigate relationships between possible predictive variables found at time T0 and alcohol consumption at time T1. At basal time (T0) the prevalence of alcohol abuse was 25.56%, whereas the prevalence one year later was 49.72% (T1). Variables that significantly predict alcohol abuse within a year are: previous alcohol abuse at T0 (p<0.001), previous abuse of drugs (p=0.011), parents attending their sporting events (p=0.005), peer problems (p=0.019), and lack of prosocial behaviour (p=0.043). In the light of our results, it can be concluded that, in adolescents, externalizing disorders seem to be determining factors of medium-term alcohol consumption.

El uso/abuso de alcohol es un problema de salud en los adolescentes. La última Encuesta sobre uso de drogas en Enseñanzas Secundarias realizada en España (ESTUDES 2014-2015), pone de manifiesto que 76,8% de los adolescentes entre 14 y 18 años consumieron alcohol en el último año y 68,2% en el último mes. El principal objetivo es determinar los factores que se asocian con el consumo de alcohol a medio plazo en una muestra de adolescentes españoles. El estudio forma parte del proyecto Saving and Empowering Young Lives in Europe (SEYLE). La muestra final estuvo compuesta por 708 estudiantes, evaluados en dos momentos temporales [basal (T0) y al año (T1)] [varones: 51,98%, edad media basal (DE)=4,43 (0,67)]. Se realizaron análisis de regresión univariante y multivariante, con el fin de investigar las relaciones entre posibles variables predictoras descritas en el momento temporal T0 y el consumo de alcohol en el momento T1.En el momento basal (T0) la prevalencia de abuso de alcohol fue del 25,56%, mientras que la prevalencia al año fue del 49,72% (T1). Las variables que predicen de forma significativa el abuso de alcohol al cabo de un año son: abuso previo del alcohol en el momento T0 (p< 0,001), abuso previo de drogas (p=0,011), padres que asisten a sus competiciones deportivas (p=0,005), problemas de relación con compañeros (p=0,019) y ausencia de comportamiento prosocial (p=0,043). A la vista de nuestros resultados se puede concluir que, en adolescentes, los trastornos externalizantes parecen ser factores determinantes de consumo de alcohol a medio plazo.

Factors associated with tobacco consumption in patients with depression. / Factores asociados con consumo de tabaco en pacientes con depresión.

Smoking and depression are related in a bidirectional way: smoking is the primary avoidable cause of illness and death in patients with depression, and depression is one of the most consistent risk factors for smoking. The main objective of this study is to investigate the relationship between smoking and depression, analyzing sociodemographic and clinical variables such as severity of symptoms, subtype of affective disorder, and its impact on suicidal behavior in the clinical population.A sample of 201 patients, over 18 years of age [mean age (SD) = 53.76 (10.36) years; women = 132 (65.7%)], with a history of depressive episode (unipolar or bipolar) or dysthymia (ICD 10 criteria) was studied.Current smoking prevalence was 43.2% and life-time prevalence 61.2%. No statistically significant differences in smoking prevalence between men and women were found (X2 = 3.896, p = 0.143). The average age of onset was 17.81 (5.60) years. There was a tendency towards a linear association between number of cigarettes/day consumed and severity of depression according to the Hamilton Depression Scale (HDRS) in current smokers (Pearson's R = 0.298, p = 0.050). Multinomial logistic regression analysis showed that current tobacco consumption was associated with higher HDRS scores, with each additional point on the HDRS increasing the likelihood of smoking by 0.062 [p = 0.032; OR (95% CI) = 1.064 (1.005-1.125)].Our results showed that depressed patients present higher prevalence of current smoking than the general population, also suggesting a relationship between severity of consumption and severity of depressive symptoms.

Tabaquismo y depresión se relacionan de forma bidireccional: el tabaquismo es la primera causa evitable de enfermedad y muerte en pacientes con depresión, y la depresión constituye uno de los factores de riesgo de tabaquismo más consistentes. El principal objetivo del presente trabajo es profundizar en la relación entre tabaquismo y depresión, analizando variables socio-demográficas y clínicas como la gravedad de los síntomas, el subtipo de trastorno afectivo, y su impacto en las conductas suicidas en población clínica.Se estudió una muestra de 201 pacientes, mayores de 18 años [edad media (SD) = 53,76 (10,36) años; mujeres = 132 (65,7%)], con historia de episodio depresivo (unipolar o bipolar) o distimia (criterios CIE 10).La prevalencia de tabaquismo actual fue 43,2% y la prevalencia vida 61,2%, no existiendo diferencias estadísticamente significativas entre hombres y mujeres (X2 = 3,896; p = 0,143). La edad media de inicio fue 17,81 (5,60) años. Se observó tendencia a asociación lineal entre número de cigarrillos/día consumidos y gravedad de la depresión según la Escala de Hamilton para la Depresión (HDRS) en los consumidores actuales de tabaco (R de Pearson = 0,298; p = 0,050). El análisis de regresión logística multinomial puso de manifiesto que el consumo actual de tabaco se asocia con puntuaciones más elevadas en la HDRS, de modo que cada incremento de un punto en dicha escala, la posibilidad de fumar aumenta en 0,062 [p = 0,032; OR (95% CI) = 1,064 (1,005-1,125)].Nuestros resultados muestran que los pacientes deprimidos presentan mayor prevalencia de consumo actual de tabaco que la población general, sugiriendo además una relación entre gravedad de consumo y gravedad de los síntomas de depresión.

Elucidating the molecular basis of MSH2-deficient tumors by combined germline and somatic analysis.

In a proportion of patients presenting mismatch repair (MMR)-deficient tumors, no germline MMR mutations are identified, the so-called Lynch-like syndrome (LLS). Recently, MMR-deficient tumors have been associated with germline mutations in POLE and MUTYH or double somatic MMR events. Our aim was to elucidate the molecular basis of MSH2-deficient LS-suspected cases using a comprehensive analysis of colorectal cancer (CRC)-associated genes at germline and somatic level. Fifty-eight probands harboring MSH2-deficient tumors were included. Germline mutational analysis of MSH2 (including EPCAM deletions) and MSH6 was performed. Pathogenicity of MSH2 variants was assessed by RNA analysis and multifactorial likelihood calculations. MSH2 cDNA and methylation of MSH2 and MSH6 promoters were studied. Matched blood and tumor DNA were analyzed using a customized next generation sequencing panel. Thirty-five individuals were carriers of pathogenic or probably pathogenic variants in MSH2 and EPCAM. Five patients harbored 4 different MSH2 variants of unknown significance (VUS) and one had 2 novel MSH6 promoter VUS. Pathogenicity assessment allowed the reclassification of the 4 MSH2 VUS and 6 probably pathogenic variants as pathogenic mutations, enabling a total of 40 LS diagnostics. Predicted pathogenic germline variants in BUB1, SETD2, FAN1 and MUTYH were identified in 5 cases. Three patients had double somatic hits in MSH2 or MSH6, and another 2 had somatic alterations in other MMR genes and/or proofreading polymerases. In conclusion, our comprehensive strategy combining germline and somatic mutational status of CRC-associated genes by means of a subexome panel allows the elucidation of up to 86% of MSH2-deficient suspected LS tumors.

Interactions of subunits Asa2, Asa4 and Asa7 in the peripheral stalk of the mitochondrial ATP synthase of the chlorophycean alga Polytomella sp.

Mitochondrial F1FO-ATP synthase of chlorophycean algae is a complex partially embedded in the inner mitochondrial membrane that is isolated as a highly stable dimer of 1600kDa. It comprises 17 polypeptides, nine of which (subunits Asa1 to 9) are not present in classical mitochondrial ATP synthases and appear to be exclusive of the chlorophycean lineage. In particular, subunits Asa2, Asa4 and Asa7 seem to constitute a section of the peripheral stalk of the enzyme. Here, we over-expressed and purified subunits Asa2, Asa4 and Asa7 and the corresponding amino-terminal and carboxy-terminal halves of Asa4 and Asa7 in order to explore their interactions in vitro, using immunochemical techniques, blue native electrophoresis and affinity chromatography. Asa4 and Asa7 interact strongly, mainly through their carboxy-terminal halves. Asa2 interacts with both Asa7 and Asa4, and also with subunit α in the F1 sector. The three Asa proteins form an Asa2/Asa4/Asa7 subcomplex. The entire Asa7 and the carboxy-terminal half of Asa4 seem to be instrumental in the interaction with Asa2. Based on these results and on computer-generated structural models of the three subunits, we propose a model for the Asa2/Asa4/Asa7 subcomplex and for its disposition in the peripheral stalk of the algal ATP synthase.

Assessing the RNA effect of 26 DNA variants in the BRCA1 and BRCA2 genes.

Comprehensive genetic testing of the breast cancer susceptibility genes BRCA1 and BRCA2 identified approximately 16% of variants of unknown significance (VUS), a significant proportion of which could affect the correct splicing of the genes. Our aim is to establish a workflow for classifying VUS in these complex genes, the first stage of which is splicing analysis. We used a combined approach consisting of five in silico splicing prediction programs and RT-PCR analysis for a set of 26 variants not previously studied at the mRNA level and six variants that had already been studied, four of which were used as positive controls as they were found to affect the splicing of these genes and the other two were used as negative controls. We identified a splicing defect in 8 of the 26 newly studied variants and ruled out splicing alteration in the remaining 18 variants. The results for the four positive and the two negative control variants were consistent with results presented in the literature. Our results strongly suggest that the combination of RNA analysis and in silico programs is an important step towards the classification of VUS. The results revealed a very high correlation between experimental data and in silico programs when using tools for predicting acceptor/donor sites but a lower correlation in the case of tools for identifying ESE elements.

Validation of a European Spanish adaptation of the Apathy Evaluation Scale-self-rated version (AES-S) in patients with schizophrenia.

INTRODUCTION: Apathy is a negative symptom of schizophrenia and is associated with poor real world functioning. Therefore, it is important to have validated psychometric instruments to assess this symptom. This is the first study to validate the Spanish adaptation of the self-rated version of the Apathy Assessment Scale (AES-S) in patients with schizophrenia. MATERIALS AND METHODS: Naturalistic, cross-sectional, validation study in 104 patients with schizophrenia evaluated using the following scales: Clinical Global Impression-Severity (CGI-S), Personal and Social Performance (PSP), Clinical Assessment Interview for Negative Symptoms (CAINS), Self-report of Negative Symptoms (SNS), Motivation and Pleasure Scale-Self-Report (MAP-SR), Calgary Depression Scale for Schizophrenia (CDSS), and Apathy Evaluation Scale-self-rated version (AES-S). RESULTS: Reliability: Internal consistency (Cronbach's alpha) was 0.908. Convergent validity: The Pearson correlation coefficient between AES-S and CAINS-MAP total scores was -0.483 (p<0.001). For SNS, total and avolition subscale scores were -0.803 and -0.639 (p<0.001), respectively. With the MAP-SR, the correlation coefficient was -0.727 (p<0.001). Divergent validity: The Pearson correlation coefficient between AES-S and PSP total scores was 0.504 (p<0.001). Furthermore, with the CDSS, the correlation coefficient was -0.431 (p<0.001). Discriminant validity: The AES-S discriminated between different levels of illness severity according to CGI-S scores. Factor analysis: A three-component solution explained 57.32% of the variance. Pearson correlations between coefficients were 1-2=0.265, 1-3=0.464, and 2-3=0.060. CONCLUSION: The Spanish AES-S is a reliable and valid instrument for assessing apathy in Spanish patients with schizophrenia. It seems to be appropriate for use in everyday clinical practice as a means of monitoring apathy in these patients.

Validation of a European Spanish-version of the Self-Evaluation of Negative Symptoms (SNS) in patients with schizophrenia.

INTRODUCTION: Negative symptoms can be grouped into five domains: apathy/avolition, anhedonia, asociality, alogia, and affective flattening. There are few validate self-rated measures that assess these five dimensions. Therefore, this study aimed to validate the Self-Evaluation of Negative Symptoms (SNS) in Spanish patients with schizophrenia. MATERIAL AND METHODS: Cross-sectional, validation study in 104 outpatients with schizophrenia evaluated using the Spanish version of the following scales: Clinical Assessment Interview for Negative Symptoms (CAINS), Positive and Negative Syndrome Scale (PANSS), Clinical Global Impression Scale for Schizophrenia (CGI-SCH), Personal and Social Performance (PSP), Motivation and Pleasure Scale - Self-Report (MAP-SR), 36-item Short-Form Health Survey (SF-36) and the Self-Evaluation of Negative Symptoms (SNS). RESULTS RELIABILITY: Internal consistency (Cronbach's alpha) was 0.915. Convergent validity: The Pearson correlation coefficient between MAP-SR and SNS Total scores was 0.660 (p<0.001). For PANSS-N, the correlation was 0.437 (p<0.005) and with the CAINS-Total was 0.478 (p<0.005). Divergent validity: The Pearson correlation coefficient between SNS and PSP was r=-0.372 (p≤0.001), and with SF-36 Physical and Mental Summary Component scores were r=-0.213 (p=0.066) and r=-0.144 (p=0.219), respectively. Discriminant validity: SNS Total scores were significantly statistically different according to the severity of the negative symptomatology rated by the CGI-SCH negative scale (p<0.001). CONCLUSION: The SNS is a reliable and valid instrument to self-rate the five domains of negative symptoms in patients with schizophrenia and seems to be appropriate for use in everyday clinical practice as a complementary measure to the evaluation performed by the clinician.