Predicting the impact of rare variants on RNA splicing in CAGI6.

Variants which disrupt splicing are a frequent cause of rare disease that have been under-ascertained clinically. Accurate and efficient methods to predict a variant's impact on splicing are needed to interpret the growing number of variants of unknown significance (VUS) identified by exome and genome sequencing. Here, we present the results of the CAGI6 Splicing VUS challenge, which invited predictions of the splicing impact of 56 variants ascertained clinically and functionally validated to determine splicing impact. The performance of 12 prediction methods, along with SpliceAI and CADD, was compared on the 56 functionally validated variants. The maximum accuracy achieved was 82% from two different approaches, one weighting SpliceAI scores by minor allele frequency, and one applying the recently published Splicing Prediction Pipeline (SPiP). SPiP performed optimally in terms of sensitivity, while an ensemble method combining multiple prediction tools and information from databases exceeded all others for specificity. Several challenge methods equalled or exceeded the performance of SpliceAI, with ultimate choice of prediction method likely to depend on experimental or clinical aims. One quarter of the variants were incorrectly predicted by at least 50% of the methods, highlighting the need for further improvements to splicing prediction methods for successful clinical application.

Gene-specific ACMG/AMP classification criteria for germline APC variants: Recommendations from the ClinGen InSiGHT Hereditary Colorectal Cancer/Polyposis Variant Curation Expert Panel.

PURPOSE: The Hereditary Colorectal Cancer/Polyposis Variant Curation Expert Panel (VCEP) was established by the International Society for Gastrointestinal Hereditary Tumours and the Clinical Genome Resource, who set out to develop recommendations for the interpretation of germline APC variants underlying Familial Adenomatous Polyposis, the most frequent hereditary polyposis syndrome. METHODS: Through a rigorous process of database analysis, literature review, and expert elicitation, the APC VCEP derived gene-specific modifications to the ACMG/AMP (American College of Medical Genetics and Genomics and Association for Molecular Pathology) variant classification guidelines and validated such criteria through the pilot classification of 58 variants. RESULTS: The APC-specific criteria represented gene- and disease-informed specifications, including a quantitative approach to allele frequency thresholds, a stepwise decision tool for truncating variants, and semiquantitative evaluations of experimental and clinical data. Using the APC-specific criteria, 47% (27/58) of pilot variants were reclassified including 14 previous variants of uncertain significance (VUS). CONCLUSION: The APC-specific ACMG/AMP criteria preserved the classification of well-characterized variants on ClinVar while substantially reducing the number of VUS by 56% (14/25). Moving forward, the APC VCEP will continue to interpret prioritized lists of VUS, the results of which will represent the most authoritative variant classification for widespread clinical use.

Comprehensive RNA and protein functional assessments contribute to the clinical interpretation of MSH2 variants causing in-frame splicing alterations.

BACKGROUND: Spliceogenic variants in disease-causing genes are often presumed pathogenic since most induce frameshifts resulting in loss of function. However, it was recently shown in cancer predisposition genes that some may trigger in-frame anomalies that preserve function. Here, we addressed this question by using MSH2, a DNA mismatch repair gene implicated in Lynch syndrome, as a model system. METHODS: Eighteen MSH2 variants, mostly localised within canonical splice sites, were analysed by using minigene splicing assays. The impact of the resulting protein alterations was assessed in a methylation tolerance-based assay. Clinicopathological characteristics of variant carriers were collected. RESULTS: Three in-frame RNA biotypes were identified based on variant-induced spliceogenic outcomes: exon skipping (E3, E4, E5 and E12), segmental exonic deletions (E7 and E15) and intronic retentions (I3, I6, I12 and I13). The 10 corresponding protein isoforms exhibit either large deletions (49-93 amino acids (aa)), small deletions (12 or 16 aa) or insertions (3-10 aa) within different functional domains. We showed that all these modifications abrogate MSH2 function, in agreement with the clinicopathological features of variant carriers. CONCLUSION: Altogether, these data demonstrate that MSH2 function is intolerant to in-frame indels caused by the spliceogenic variants analysed in this study, supporting their pathogenic nature. This work stresses the importance of combining complementary RNA and protein approaches to ensure accurate clinical interpretation of in-frame spliceogenic variants.

Warmer water, high light intensity, lithium and microplastics: Dangerous environmental combinations to zooplankton and Global Health?

Nowadays there is a high concern about the combined effects of global warming and emerging environmental contaminants with significant increasing trends of use, such as lithium (Li) and microplastics (MPs), both on wildlife and human health. Therefore, the effects of high light intensity (26,000 lx) or warmer water temperature (25 °C) on the long-term toxicity of Li and mixtures of Li and MPs (Li-MPs mixtures) were investigated using model populations of the freshwater zooplankton species Daphnia magna. Three 21-day bioassays were done in the laboratory at the following water temperatures and light intensities: (i) 20 °C/10830 lx; (ii) 20 °C/26000 lx (high light intensity); (iii) 25 °C/10830 lx (warmer temperature). Based on the 21-day EC50s on reproduction, high light intensity increased the reproductive toxicity of Li and Li-MPs mixtures by ~1.3 fold; warmer temperature increased the toxicity of Li by ~1.2 fold, and the toxicity of Li-MPs mixtures by ~1.4 fold based on the concentration of Li, and by ~2 fold based on the concentrations of MPs. At high light intensity, Li (0.04 mg/L) and Li-MPs mixtures (0.04 Li + 0.09 MPs mg/L) reduced the population fitness by 32 % and 41 %, respectively. Warmer temperature, Li (0.05 mg/L) and Li-MPs mixtures (0.05 Li + 0.09 MPs mg/L) reduced it by 63 % and 71 %, respectively. At warmer temperature or high light intensity, higher concentrations of Li and Li-MPs mixtures lead to population extinction. Based on the population growth rate and using data of bioassays with MPs alone done simultaneously, Li and MPs interactions were antagonistic or synergistic depending on the scenario. High light intensity and chemical stress generally acted synergistically. Warmer temperature and chemical stress always acted synergistically. These findings highlight the threats of long-term exposure to Li and Li-MPs mixtures to freshwater zooplankton and Global Health in a warmer world.

Global Real-World Evidence of Sofosbuvir/Velpatasvir as a Highly Effective Treatment and Elimination Tool in People with Hepatitis C Infection Experiencing Mental Health Disorders.

Hepatitis C virus (HCV) is prevalent in people with mental health disorders, a priority population to diagnose and cure in order to achieve HCV elimination. This integrated analysis pooled data from 20 cohorts in seven countries to evaluate the real-world effectiveness of the pangenotypic direct-acting antiviral (DAA) sofosbuvir/velpatasvir (SOF/VEL) in people with mental health disorders. HCV-infected patients diagnosed with mental health disorders who were treated with SOF/VEL for 12 weeks without ribavirin as part of routine clinical practice were included. The primary outcome was sustained virological response (SVR) in the effectiveness population (EP), defined as patients with an available SVR assessment. Secondary outcomes were reasons for not achieving SVR, characteristics of patients with non-virological failures, adherence, and time from HCV RNA diagnosis to SOF/VEL treatment initiation. A total of 1209 patients were included; 142 did not achieve an SVR for non-virological reasons (n = 112; 83 lost to follow-up, 20 early treatment discontinuations) or unknown reasons (n = 30). Of the 1067 patients in the EP, 97.4% achieved SVR. SVR rates in the EP were ≥95% when stratified by type of mental health disorder and other complicating baseline characteristics, including active injection drug use and antipsychotic drug use. Of 461 patients with data available in the EP, only 2% had an adherence level < 90% and 1% had an adherence level < 80%; all achieved SVR. Patients with mental health disorders can be cured of HCV using a well-tolerated, pangenotypic, protease inhibitor-free SOF/VEL regimen. This DAA allows the implementation of a simple treatment algorithm, with minimal monitoring requirements and fewer interactions with central nervous system drugs compared with protease-inhibitor DAA regimens.

SPiP: Splicing Prediction Pipeline, a machine learning tool for massive detection of exonic and intronic variant effects on mRNA splicing.

Modeling splicing is essential for tackling the challenge of variant interpretation as each nucleotide variation can be pathogenic by affecting pre-mRNA splicing via disruption/creation of splicing motifs such as 5'/3' splice sites, branch sites, or splicing regulatory elements. Unfortunately, most in silico tools focus on a specific type of splicing motif, which is why we developed the Splicing Prediction Pipeline (SPiP) to perform, in one single bioinformatic analysis based on a machine learning approach, a comprehensive assessment of the variant effect on different splicing motifs. We gathered a curated set of 4616 variants scattered all along the sequence of 227 genes, with their corresponding splicing studies. The Bayesian analysis provided us with the number of control variants, that is, variants without impact on splicing, to mimic the deluge of variants from high-throughput sequencing data. Results show that SPiP can deal with the diversity of splicing alterations, with 83.13% sensitivity and 99% specificity to detect spliceogenic variants. Overall performance as measured by area under the receiving operator curve was 0.986, better than SpliceAI and SQUIRLS (0.965 and 0.766) for the same data set. SPiP lends itself to a unique suite for comprehensive prediction of spliceogenicity in the genomic medicine era. SPiP is available at: https://sourceforge.net/projects/splicing-prediction-pipeline/.

Quality of working life of professors in times of social distancing.

Introduction: Quality of working life consists in a dynamic management of physical, technological, and sociopsychological factors that affect culture and renovate the organizational climate, reflecting on workers' wellbeing. Objectives: To identify changes in eating habits, physical activity, and hours of sleep, as well as their influence on the quality of working life of professors who shifted to remote working during the pandemic. Methods: This study has a quantitative, descriptive, observational design and was performed with faculty members of a private higher education institution in May 2020; 40 professors participated in our research. We used a virtual questionnaire containing Walton's quality of working life scale (1973) and questions on lifestyle habits. Results: We observed that changes in lifestyle habits were correlated with lower or higher perceptions of satisfaction with the 4 evaluated criteria referring to quality of life: working conditions, use of capacities at work, social interaction at work, and the space work occupies in life. Conclusions: The mean quality of working life score of faculty members working remotely during lockdown demonstrated that, despite changes in work and lifestyle habits, professors still maintained a certain degree of satisfaction with the aspects of work evaluated in this study.

Splicing analyses for variants in MMR genes: best practice recommendations from the European Mismatch Repair Working Group.

Over 20% of the DNA mismatch repair (MMR) germline variants in suspected Lynch syndrome patients are classified as variants of uncertain significance (VUS). Well-established functional assays are pivotal for assessing the biological impact of these variants and provide relevant evidence for clinical classification. In our collaborative European Mismatch Repair Working Group (EMMR-WG) we compared three different experimental approaches for evaluating the effect of seven variants on mRNA splicing in MMR genes: (i) RT-PCR of full-length transcripts (FLT), (ii) RT-PCR of targeted transcript sections (TTS), both from patient biological samples and (iii) minigene splicing assays. An overall good concordance was observed between splicing patterns in TTS, FLT and minigene analyses for all variants. The FLT analysis depicted a higher number of different isoforms and mitigated PCR-bias towards shorter isoforms. TTS analyses may miss aberrant isoforms and minigene assays may under/overestimate the severity of certain splicing defects. The interpretation of the experimental findings must be cautious to adequately discriminate abnormal events from physiological complex alternative splicing patterns. A consensus strategy for investigating the impact of MMR variants on splicing was defined. First, RNA should be obtained from patient's cell cultures (such as fresh lymphocyte cultures) incubated with/without a nonsense-mediated decay inhibitor. Second, FLT RT-PCR analysis is recommended to oversee all generated isoforms. Third, TTS analysis and minigene assays are useful independent approaches for verifying and clarifying FLT results. The use of several methodologies is likely to increase the strength of the experimental evidence which contributes to improve variant interpretation.

Long-term effects of lithium and lithium-microplastic mixtures on the model species Daphnia magna: Toxicological interactions and implications to 'One Health'.

Environmental contamination with lithium (Li) and microplastics (MP) has been steadily increasing and this trend is expected to continue in the future. Many freshwater ecosystems, which are crucial to reach the United Nations Sustainable Development Goals, are particularly vulnerable to Li and MP contamination, and other pressures. The long-term effects of Li, either alone or combined with MP (Li-MP mixtures), were investigated using the freshwater zooplankton micro-crustacean Daphnia magna as model species. In the laboratory, D. magna females were exposed for 21 days to water concentrations of Li (0.02, 0.04, 0.08 mg/L) or Li-MP mixtures (0.02 Li + 0.04 MP, 0.04 Li + 0.09 MP mg/L, 0.08 Li + 0.19 MP mg/L). In the range of concentrations tested, Li and Li-MP mixtures caused parental mortality, and decreased the somatic growth (up to 20% and 40% reduction, respectively) and the reproductive success (up to 93% and 90% reduction, respectively). The 21-day EC50s of Li and Li-MP mixtures on D. magna reproduction were 0.039 mg/L and 0.039 Li + 0.086 MP mg/L, respectively. Under exposure to the highest concentration of Li (0.08 mg/L) and Li-MP mixtures (0.08 Li + 0.19 MP mg/L), the mean of D. magna population growth rate was reduced by 67% and 58%, respectively. Based on the population growth rate and using data from a bioassay testing the same concentrations of MP alone and carried simultaneously, the toxicological interaction between Li and MP was antagonism under exposure to the lowest and the highest concentrations of Li-MP mixtures, and synergism under exposure to the medium concentration of Li-MP mixtures. These findings highlight the need of further investigating the combined effects of contaminants, and the threat of long-term environmental contamination with Li and MP to freshwater zooplankton, biodiversity, ecosystem services and 'One Health'.

Risk factors for the emergence of multidrug-resistant organisms in liver cirrhosis / Factores de riesgo para la aparición de organismos multirresistentes en la cirrosis hepática

Background: Multidrug-resistant organisms (MDROs) are a reality that can alter the paradigm of treatment and prevention of infection in patients with liver cirrhosis (LC).ObjectiveIdentify risk factors for the occurrence of MDROs in patients with LC.Patients and methods: Prospective study from October 2017 to March 2018 in consecutively hospitalized patients with decompensated LC with infection. Blood, urine and ascitic fluid cultures were analyzed. A p-value ≤0.05 was considered statistically significant. Results: MDROs isolated in 18 of 52 episodes of infection. MDROs were associated with the use of proton pump inhibitors (PPIs) (p=0.0312), antibiotic therapy in the last 90 days (p=0.0033) and discharge within preceding 30 days or current hospitalization above 48h (p=0.0082). There was higher 90-day mortality in patients with MDROs infection (71.4% versus 35.7%, p=0.0316).Conclusion: MDROs infections were prevalent in this cohort and associated with 90-day mortality. Use of PPIs and antibiotics increased the risk of MDROs infections, suggesting that its prescription should be restricted to formal indication. Hospitalization was associated with the onset of MDROs, so LC patients should stay at the hospital the least possible. It is relevant to investigate other factors predisposing to the emergence of these microorganisms, in order to prevent it. (AU)

Antecedentes: Los organismos multirresistentes (MDROs, por sus siglas en inglés) son una realidad que puede alterar el paradigma del tratamiento y la prevención de la infección en los pacientes con cirrosis hepática (LC, por sus siglas en inglés).Objetivo: Identificar los factores de riesgo para la aparición de MDROs en pacientes con LC.Pacientes y métodos: Estudio prospectivo de octubre de 2017 a marzo de 2018 en pacientes hospitalizados consecutivamente con LC descompensada con infección. Se analizaron los cultivos de sangre, orina y líquido ascítico. Se consideró estadísticamente significativo un valor de p≤0,05.Resultados: Se aislaron MDROs en 18 de los 52 episodios de infección. Los MDROs se asociaron con el uso de inhibidores de la bomba de protones (IBP) (p=0,0312), la terapia antibiótica en los últimos 90 días (p=0,0033) y el alta en los 30 días anteriores o la hospitalización actual superior a 48h (p=0,0082). Hubo una mayor mortalidad a los 90 días en los pacientes con infección por MDROs (71,4 frente al 35,7%; p=0,0316).Conclusión: Las infecciones por MDROs fueron prevalentes en esta cohorte, y se asociaron con la mortalidad a los 90 días. El uso de IBP y antibióticos aumentó el riesgo de infecciones por MDROs, lo que sugiere que su prescripción debe restringirse a la indicación formal. La hospitalización se asoció a la aparición de MDROs, por lo que los pacientes con LC deberían permanecer en el hospital el menor tiempo posible. Es relevante investigar otros factores que predisponen a la aparición de estos microorganismos para prevenirla. (AU)

Indicators for local health plan monitoring and evaluation: A modified Delphi consensus.

OBJECTIVE: There is still a lack of health indicators for monitoring and evaluating health planning at the local level. In Portugal, local health plans (LHP) include a prioritized set of health priorities, which should be monitored and evaluated. This study is an example of a low-resource method to identify and reuse indicators for LHP monitoring and evaluation already collected for other purposes. DESIGN AND SAMPLE: A modified Delphi consensus method was applied, with three rounds of email rating questionnaires and a final meeting, between January 2018 and January 2019. The Delphi panel consisted of eight members from the Planning and Administration Group of the Espinho/Gaia Local Public Health Unit. MEASUREMENTS: Panelists were asked to assess the indicators' validity for monitoring diseases/determinants from a pre-selected list of potential binomials between 140 PHC indicators and 15 diseases/determinants. RESULTS: After four rounds, there was consensus in considering 141 binomials (34.0%) as appropriate, diabetes mellitus being the disease with more appropriate indicators. CONCLUSION: This study portrays the applicability of a commonly used, easy and low-resource method in a Portuguese Local Public Health Unit to select and reuse primary health care indicators for LHP monitoring and evaluation.

Risk factors for the emergence of multidrug-resistant organisms in liver cirrhosis.

BACKGROUND: Multidrug-resistant organisms (MDROs) are a reality that can alter the paradigm of treatment and prevention of infection in patients with liver cirrhosis (LC). OBJECTIVE: Identify risk factors for the occurrence of MDROs in patients with LC. PATIENTS AND METHODS: Prospective study from October 2017 to March 2018 in consecutively hospitalized patients with decompensated LC with infection. Blood, urine and ascitic fluid cultures were analyzed. A p-value ≤0.05 was considered statistically significant. RESULTS: MDROs isolated in 18 of 52 episodes of infection. MDROs were associated with the use of proton pump inhibitors (PPIs) (p=0.0312), antibiotic therapy in the last 90 days (p=0.0033) and discharge within preceding 30 days or current hospitalization above 48h (p=0.0082). There was higher 90-day mortality in patients with MDROs infection (71.4% versus 35.7%, p=0.0316). CONCLUSION: MDROs infections were prevalent in this cohort and associated with 90-day mortality. Use of PPIs and antibiotics increased the risk of MDROs infections, suggesting that its prescription should be restricted to formal indication. Hospitalization was associated with the onset of MDROs, so LC patients should stay at the hospital the least possible. It is relevant to investigate other factors predisposing to the emergence of these microorganisms, in order to prevent it.

Microplastics in fishes from an estuary (Minho River) ending into the NE Atlantic Ocean.

Wild fish (Cyprinus carpio, Mugil cephalus, Platichthys flesus) from an estuary of the NE Atlantic coast were investigated for plastic contamination (N = 128). From the 1289 particles recovered from fish samples, 883 were plastics. Among these, 84% were fibres and 97% were microplastics. Thirty-six polymers were identified. The number of microplastics (mean ± SD) per individual fish (MP/fish) was 8 ± 6 in C. carpio, 10 ± 9 in M. cephalus and 2 ± 2 in P. flesus. The means of MP/fish per body site were 6 ± 7 in gastrointestinal tract, 0.5 ± 1.1 in gills, 0.3 ± 0.7 in liver and 0.6 ± 1.2 in muscle samples. A few large fibres in liver (≤ 4841 µm) and muscle (≤ 5810 µm) samples were found. The results evidence the existence of high fish contamination by microplastics and reinforce the need of further research on plastic pollution in estuaries.

Perfil metacognitivo de estudantes de enfermagem e desempenho acadêmico / Metacognitive profile of nursing students and academic performance / Perfil metacognitivo de estudiantes de enfermería y rendimiento académico

Objetivo: identificar o perfil metacognitivo de ingressantes na graduação em enfermagem e seu potencial de predizer o desempenho acadêmico durante o primeiro semestre letivo. Método: estudo observacional longitudinal prospectivo, descritivo e explicativo. Utilizou-se o Inventário de Consciência Metacognitiva de Schraw e Dennison para identificar características do perfil metacognitivo dos ingressantes e as notas finais alcançadas nas disciplinas cursadas no primeiro semestre como indicativo do desempenho acadêmico. Resultados: os ingressantes eram predominantemente mulheres, com menos de 21 anos, solteiras, autodeclaradas pardas e com alguma forma de bolsa de estudo. A habilidade metacognitiva de Estratégia de Correção foi a que os estudantes mais colocaram em prática. Observou-se correlação (p<0,05 e r=0,2844) entre a habilidade de Planejamento e a nota de aproveitamento final. Conclusão: a habilidade de Planejamento foi a única com potencial de predizer melhor desempenho acadêmico, embora ela não seja a habilidade mais praticada pelos estudantes.(AU)

Objective: to identify the metacognitive profile of newcomers in nursing undergraduate courses and their potential to predict academic performance during the first academic semester. Method: prospective, descriptive and explanatory longitudinal observational study. We used the Schraw and Dennison's Metacognitive Awareness Inventory to identify characteristics of the metacognitive profile of the students and the final grades achieved in the subjects taken in the first semester as an indication of academic performance. Results: the newcomers were predominantly women, under 21 years, single, self-declared brown and with some form of scholarship. The Correction Strategy metacognitive skill was the most put into practice by the surveyed students. We noted a correlation (p<0.05 and r=0.2844) between the Planning skill and the final yield grade. Conclusion: the Planning skill was the only one with the potential to predict better academic performance, although it is not the skill most practiced by students.(AU)

Objetivo: identificar el perfil metacognitivo de los principiantes en cursos de pregrado en enfermería y su potencial para predecir el rendimiento académico durante el primer semestre lectivo. Método: estudio observacional longitudinal prospectivo, descriptivo y explicativo. Se utilizó el Inventario de Conciencia Metacognitiva de Schraw y Dennison para identificar las características del perfil metacognitivo de los principiantes y las calificaciones finales obtenidas en las materias cumplidas en el primer semestre como indicador del rendimiento académico. Resultados: los participantes eran predominantemente mujeres, menores de 21 años, solteras, autodeclaradas pardas y con algún tipo de beca. La habilidad metacognitiva de la Estrategia de Corrección fue la que la mayoría de los estudiantes pusieron en práctica. Se notó una correlación (p<0.05 y r=0.2844) entre la habilidade de Planificación y la calificación de rendimiento final. Conclusión: la habilidad de Planificación fue la única con potencial para predecir un mejor rendimiento académico, aunque no es la habilidad más practicada por los estudiantes.(AU)

Predictors for incomplete response to ursodeoxycholic acid in primary biliary cholangitis. Data from a national registry of liver disease.

BACKGROUND: The current standard of treatment in primary biliary cholangitis (PBC) is ursodeoxycholic acid (UDCA), although a considerable proportion of patients show incomplete response resulting in disease progression. OBJECTIVE: This study aimed to assess the prevalence of incomplete response to UDCA and determine associated patients' characteristics. METHODS: Patients with PBC as main diagnosis were included from a national multicentric patient registry-Liver.pt. Main endpoints included incomplete response to UDCA treatment according to Barcelona, Paris I and Paris II criteria, Globe and UK PBC scores and the association between baseline characteristics and incomplete response according to Paris II criteria. RESULTS: A total of 434 PBC patients were identified, with a mean age of 55 years and 89.2% females. Nearly half of patients were asymptomatic at diagnosis and 93.2% had positive anti-mitochondrial antibodies. Almost all patients (95.6%) had been prescribed at least one drug for PBC treatment. At the last follow-up visit, 93.3% were under treatment of which 99.8% received UDCA. Incomplete response to UDCA was observed in 30.7%, 35.3%, 53.7% and 36.4% of patients according to Barcelona, Paris I, Paris II criteria and Globe score, respectively. After adjusting for age and sex, and accordingly to Paris II criteria, the risk for incomplete biochemical response was 25% higher for patients with cirrhosis at diagnosis (odds ratio [OR] = 1.25; 95% confidence interval [95%CI]: 1.02-1.54; p = 0.033) and 35% (95%CI:1.06-1.72; p = 0.016) and 5% (OR = 1.05; 95%CI:1.01-1.10; p = 0.013) for those with elevated gamma-glutamyl transferase (GGT) and alkaline phosphatase (ALP). CONCLUSION: A considerable proportion of patients showed incomplete biochemical response to UDCA treatment according to Paris II criteria. Cirrhosis, elevated GGT and ALP at diagnosis were identified as associated risk factors for incomplete response. Early identification of patients at risk of incomplete response could improve treatment care and guide clinical decision to a more careful patient monitorization.

Long-term adverse effects of microplastics on Daphnia magna reproduction and population growth rate at increased water temperature and light intensity: Combined effects of stressors and interactions.

In many ecosystems, the zooplankton community has been pressured simultaneously by microplastic pollution and alterations resulting from global climate changes. The potential influence of light intensity rise (from 10,830 lx to 26,000 lx) and water temperature rise (from 20 °C to 25 °C) on the long term-toxicity of microplastics (MPs) to Daphnia magna were investigated. Three 21-day laboratory bioassays with model MPs (1-5 µm diameter) were carried out at (i) 20 °C/10830 lx, (ii) 20 °C/26000 lx, and (iii) 25 °C/10830 lx. In each bioassay, one control (no MPs) and three MP concentrations (0.04, 0.09, 0.19 mg/L) were tested. In all the bioassays, MPs caused parental and juvenile mortality, and reduced the somatic growth, reproduction and population growth rate. The MP EC50s on living offspring (95% confidence interval within brackets) were 0.146 mg/L (0.142-0.151 mg/L) at 20 °C/10830 lx, 0.102 mg/L (0.099-0.105 mg/L) at 20 °C/26000 lx, and 0.101 mg/L (0.098-0.104 mg/L) at 25 °C/10830 lx. Relatively to the respective control group, 0.19 mg/L of MPs decreased the mean of the population growth rate by 27% at 20 °C/10830 lx, 38% at 20 °C/26000 lx and 59% at 25 °C/10830 lx. Based on the population growth rate and in relation to 20 °C/10830 lx (control, no MPs), the interaction between increased light intensity (26,000 lx) and MPs was synergism (at all the MP concentrations tested). The interaction between water temperature rise (25 °C) and MPs was antagonism at 0.04 mg/L of MPs and synergism at 0.09 and 0.19 mg/L of MPs. In the present scenario of climate changes and global MP pollution such findings raise high concern because zooplankton communities are crucial for aquatic biodiversity conservation, ecosystem functioning and services provided to humans. Further studies on the combined effects of MPs, other common pollutants, and alterations due to climate changes are needed.

Functional characterization of ABCC8 variants of unknown significance based on bioinformatics predictions, splicing assays, and protein analyses: Benefits for the accurate diagnosis of congenital hyperinsulinism.

ABCC8 encodes the SUR1 subunit of the ß-cell ATP-sensitive potassium channel whose loss of function causes congenital hyperinsulinism (CHI). Molecular diagnosis is critical for optimal management of CHI patients. Unfortunately, assessing the impact of ABCC8 variants on RNA splicing remains very challenging as this gene is poorly expressed in leukocytes. Here, we performed bioinformatics analysis and cell-based minigene assays to assess the impact on splicing of 13 ABCC8 variants identified in 20 CHI patients. Next, channel properties of SUR1 proteins expected to originate from minigene-detected in-frame splicing defects were analyzed after ectopic expression in COSm6 cells. Out of the analyzed variants, seven induced out-of-frame splicing defects and were therefore classified as recessive pathogenic, whereas two led to skipping of in-frame exons. Channel functional analysis of the latter demonstrated their pathogenicity. Interestingly, the common rs757110 SNP increased exon skipping in our system suggesting that it may act as a disease modifier factor. Our strategy allowed determining the pathogenicity of all selected ABCC8 variants, and CHI-inheritance pattern for 16 out of the 20 patients. This study highlights the value of combining RNA and protein functional approaches in variant interpretation and reveals the minigene splicing assay as a new tool for CHI molecular diagnostics.

The relationship between dyadic coping and dyadic adjustment among HIV-serodiscordant couples.

Living within an HIV-serodiscordant relationship has been recognized as a stressful experience for both HIV-infected and HIV-uninfected partners. However, no study has examined the association between dyadic coping (DC) and dyadic adjustment of such couples. In this study, we analysed the association between DC (positive, negative, and common DC) and dyadic adjustment (consensus, satisfaction, cohesion) among HIV-serodiscordant couples, considering individual and cross-partner effects. This cross-sectional study included a sample of 44 HIV-serodiscordant different-sex couples, in a relationship for an average of 16.46 years. The self-reported measures included the Dyadic Coping Inventory and the Revised Dyadic Adjustment Scale. For HIV-infected partners, their own common DC was significantly associated with cohesion, and a cross-partner effect of common DC on satisfaction was found. For HIV-uninfected partners, individual effects of common DC on all dyadic adjustment subscales and a cross-partner effect of common DC on cohesion were found. Additionally, their own and their HIV-infected partners' negative DC were significantly associated with cohesion and satisfaction, respectively. These findings suggest that the perception of common DC has a particularly important role in explaining the different components of dyadic adjustment of both partners facing HIV-serodiscordancy, whereas negative DC is linked to the adjustment of HIV-uninfected partners.

Interprofessional simulation effects for healthcare students: A systematic review and meta-analysis.

OBJECTIVE: To conduct a systematic review of studies describing the effects of interprofessional education (IPE) on collaborative competence using simulated-based training of undergraduate healthcare students. DESIGN: A systematic review and meta-analysis based on PRISMA guidelines. DATA SOURCES: PubMed and Cumulative Index to Nursing and Allied Health Literature databases were searched to identify articles in all languages published up to 2018. The systematic review protocol was registered at PROSPERO under number 133330. REVIEW METHODS: In total, 419 articles were identified. The following articles were excluded: non-English articles, articles for which the full text was not available, articles that did not employ a validated tool, articles that did not use quasi-experimental methods and that did not assess healthcare student populations. Eleven studies were included, and 6 were submitted to meta-analysis using forest plots through RevMan 5.3. RESULTS: Interprofessional simulation analysis yielded results regarding participants, protocols, scenarios, validated tools, collaborative competencies and primary outcomes. The meta-analysis was organized based on assessment tool, and summary value, confidence interval, and Z test results for the random-effects model are presented. CONCLUSION: Quantitative analysis reveals a positive impact and the effectiveness of interprofessional simulation. However, more research should be conducted utilizing clinical trials with distinguished analyses for each collaborative competency factor to assess long-term effects on the outcome.