RESUMO
Identification of qualitative variants of von Willebrand disease (VWD) can be a diagnostic challenge because of discrepant results obtained in the multiple laboratory tests available for its appropriate classification. We report two cases of infrequent inherited variants of VWD with unclear preliminary results with the test panel available at the time of first consultation and that were finally diagnosed as a VWD type 2A/IID with a c.8318 G > C, p.Cys2773Ser mutation and a VWD type 2M with c.4225 T > G, p.Val1409Phe mutation, respectively. The description of these two cases highlights that despite the limited diagnostic panel for the evaluation of von Willebrand Factor (VWF) functionality, the multimeric analysis and genetic family studies were fundamental tools to achieve the final diagnosis.
Assuntos
Doenças de von Willebrand/diagnóstico , Adulto , Feminino , Humanos , Pessoa de Meia-Idade , Adulto JovemAssuntos
Psoríase/genética , Receptor 3 Toll-Like/genética , Receptor 7 Toll-Like/genética , Receptor Toll-Like 9/genética , Adulto , Idade de Início , Idoso , Estudos de Casos e Controles , Feminino , Seguimentos , Predisposição Genética para Doença , Humanos , Masculino , Pessoa de Meia-Idade , Polimorfismo de Nucleotídeo Único , Psoríase/diagnóstico , Fatores de Risco , Índice de Gravidade de DoençaRESUMO
Essentials The Royal disease (RD) is a form of hemophilia B predicted to be caused by a splicing mutation. We generated an iPSC-based model of the disease allowing mechanistic studies at the RNA level. F9 mRNA analysis in iPSC-derived hepatocyte-like cells showed the predicted abnormal splicing. Mutated F9 mRNA level was very low but we also found traces of wild type transcripts. SUMMARY: Background The royal disease is a form of hemophilia B (HB) that affected many descendants of Queen Victoria in the 19th and 20th centuries. It was found to be caused by the mutation F9 c.278-3A>G. Objective To generate a physiological cell model of the disease and to study F9 expression at the RNA level. Methods Using fibroblasts from skin biopsies of a previously identified hemophilic patient bearing the F9 c.278-3A>G mutation and his mother, we generated induced pluripotent stem cells (iPSCs). Both the patient's and mother's iPSCs were differentiated into hepatocyte-like cells (HLCs) and their F9 mRNA was analyzed using next-generation sequencing (NGS). Results and Conclusion We demonstrated the previously predicted aberrant splicing of the F9 transcript as a result of an intronic nucleotide substitution leading to a frameshift and the generation of a premature termination codon (PTC). The F9 mRNA level in the patient's HLCs was significantly reduced compared with that of his mother, suggesting that mutated transcripts undergo nonsense-mediated decay (NMD), a cellular mechanism that degrades PTC-containing mRNAs. We also detected small proportions of correctly spliced transcripts in the patient's HLCs, which, combined with genetic variability in splicing and NMD machineries, could partially explain some clinical variability among affected members of the European royal families who had lifespans above the average. This work allowed the demonstration of the pathologic consequences of an intronic mutation in the F9 gene and represents the first bona fide cellular model of HB allowing the study of rare mutations at the RNA level.
Assuntos
Fator IX/genética , Hemofilia B/genética , Hepatócitos/metabolismo , Células-Tronco Pluripotentes Induzidas/metabolismo , Mutação , RNA Mensageiro/genética , Adolescente , Processamento Alternativo , Diferenciação Celular , Linhagem Celular , Fator IX/metabolismo , Feminino , Predisposição Genética para Doença , Hemofilia B/sangue , Hemofilia B/diagnóstico , Sequenciamento de Nucleotídeos em Larga Escala , Humanos , Masculino , Fenótipo , RNA Mensageiro/metabolismo , Análise de Sequência de RNAAssuntos
Assistência Ambulatorial/métodos , Assistência Ambulatorial/psicologia , Antineoplásicos/administração & dosagem , Bombas de Infusão , Linfoma não Hodgkin/tratamento farmacológico , Adulto , Idoso , Feminino , Humanos , Linfoma não Hodgkin/enfermagem , Masculino , Pessoa de Meia-Idade , Satisfação do Paciente , Inquéritos e QuestionáriosRESUMO
OBJECTIVE: To evaluate the relationship between unemployment and mental health problems. DESIGN: Crossover study, using systematic sampling of medical records. SETTING: Health District in the province of Barcelona. PATIENTS AND OTHER PARTICIPANTS: 189 medical records from 1,549 unemployed people, and another 189 paired for age and gender of people in work, were included in the study. MEASUREMENTS AND MAIN RESULTS: The unemployed group had between 1.3% and 18.8% more people with mental health problems than the in-work group. There were no significant differences between the two groups as to their use of primary care health services. CONCLUSIONS: It was established that there is a connection between unemployment and mental health problems. Prospective studies to follow cases might contribute more data on the cause and effect relationship.
