CARATkids questionnaire development process

Introdução: A asma e a rinite alérgica (ARA) são doenças inflamatórias crónicas das vias aéreas que frequentemente coexistem. O questionário para avaliação do controlo da asma e da rinite alérgica (CARAT) encontra‑se validado para o adulto. O objectivo foi desenvolver o questionário CARATkids para crianças entre os 4 e os 12 anos de idade, com o diagnóstico médico de ARA. Este trabalho pretende descrever o processo do desenvolvimento do questionário. Métodos: O desenvolvimento do questionário foi estruturado em três fases: 1) revisão da literatura sobre questionários pediátricos existentes; 2) realização de reuniões de consenso que permitiu a criação de uma versão preliminar do CARATkids, com duas versões (crianças e pais), composto por 17 itens. Para as crianças, as perguntas foram acompanhadas por ilustrações para cada questão, com formato de resposta dicotómica (sim / não). Para os pais o questionário manteve o formato de CARAT17, com questões com 4 pontos de Likert e outras questões com opção de resposta com escala dicotómica; 3) realização de um estudo transversal através de entrevistas cognitivas efectuadas a 29 crianças e respectivos pais. Resultados: Foram incluídas 29 crianças (11 do sexo feminino) e respectivos pais. A mediana de idades (P25‑P75) foi de 8 (6‑10) anos. As crianças com 4 a 5 anos não sabiam ler o questionário; com 6‑8 eram capazes de ler / compreender as perguntas, embora referindo dificuldades para algumas expressões. As crianças com mais de 9 anos consideraram o questionário muito simples e claro. O grau de concordância entre as crianças e seus pais foi de 61%, tendo ambos considerado as ilustrações muito claras e esclarecedoras quanto aos conceitos subjacentes. Os pais concordaram que a versão destinada aos pais estava muito completa, considerando a versão da criança muito clara e adequada. Consideraram ainda a escala dicotómica como mais apropriada para crianças, em comparação com a escala de Likert do questionário aplicado aos pais. As expressões identificadas como de difícil entendimento foram alteradas. Conclusão: O questionário CARATkids é o primeiro que avalia o controlo da asma e da rinite na criança. Os testes cognitivos demonstraram a sua aplicabilidade dos 6 aos 12 anos de idade.info:eu-repo/semantics/publishedVersio

Development Process and Cognitive Testing of CARATkids - Control of Allergic Rhinitis and Asthma Test for Children

Background: Allergic rhinitis and asthma (ARA) are chronic inflammatory diseases of the airways that often coexist in children. The only tool to assess the ARA control, the Control of Allergic Rhinitis and Asthma Test (CARAT) is to be used by adults. We aimed to develop the Pediatric version of Control of Allergic Rhinitis and Asthma Test (CARATkids) and to test its comprehensibility in children with 4 to 12 years of age. Methods: The questionnaire development included a literature review of pediatric questionnaires on asthma and/or rhinitis control and two consensus meetings of a multidisciplinary group. Cognitive testing was carried out in a cross-sectional qualitative study using cognitive interviews. Results: Four questionnaires to assess asthma and none to assess rhinitis control in children were identified. The multidisciplinary group produced a questionnaire version for children with 17 questions with illustrations and dichotomous (yes/no) response format. The version for caregivers had 4-points and dichotomous scales. Twenty-nine children, 4 to 12 years old, and their caregivers were interviewed. Only children over 6 years old could adequately answer the questionnaire. A few words/expressions were not fully understood by children of 6 to 8 years old. The drawings illustrating the questions were considered helpful by children and caregivers. Caregivers considered the questionnaire complete and clear and preferred dichotomous over the 4-points scales. The proportion of agreement between children and their caregivers was 61%. The words/expressions that were difficult to understand were amended. Conclusion: CARATkids, the first questionnaire to assess a child’s asthma and rhinitis control was developed and its content validity was assured. Cognitive testing showed that CARATKids is well-understood by children 6 to 12 years old. The questionnaire’s measurement properties can now be assessed in a validation study

Triple Regimen with Rituximab, Plasmapheresis and Intravenous Immunoglobulin in the Treatment of Dialysis Dependent Acute Humoral-Mediated Rejection in Kidney Grafts

Introduction: The clinical importance of humoral-mediated acute rejection has been progressively recognised. Early recognition and treatment with plasmapheresis and intravenous immunoglobulin have recently improved short term prognosis. Case report: In this report we describe the clinical features of three 2nd transplant patients developing severe acute humoral rejection during the first week post-transplant while on anti-thymocyte globulin therapy. Treatment with plasmapheresis/ intravenous immunoglobulin/rituximab resulted in rapid reversal of oliguria,and recovery of renal function within the 1st week of treatment in 2/3 patients. Diagnosis was confirmed by graft biopsies revealing peritubular neutrophiles and C4d deposits. Sequential graft biopsies in all three patients revealed complete histological recovery within two weeks. One patient never recovered renal function, and one patient lost his graft at three months following hemorrhagic shock. After 2 years follow up, the remaining patient maintains a serum creatinine of 1.1mg/dl. Conclusion: The regimen using plasmapheresis plus intravenous immunoglobulin and rituximab was effective in rapidly reversing severe acute humoral rejection

Uridylation and adenylation of RNAs

The posttranscriptional addition of nontemplated nucleotides to the 3′ ends of RNA molecules can have a significant impact on their stability and biological function. It has been recently discovered that nontemplated addition of uridine or adenosine to the 3′ ends of RNAs occurs in different organisms ranging from algae to humans, and on different kinds of RNAs, such as histone mRNAs, mRNA fragments, U6 snRNA, mature small RNAs and their precursors etc. These modifications may lead to different outcomes, such as increasing RNA decay, promoting or inhibiting RNA processing, or changing RNA activity. Growing pieces of evidence have revealed that such modifications can be RNA sequence-specific and subjected to temporal or spatial regulation in development. RNA tailing and its outcomes have been associated with human diseases such as cancer. Here, we review recent developments in RNA uridylation and adenylation and discuss the future prospects in this research area

Structural basis for acceptor RNA substrate selectivity of the 3' terminal uridylyl transferase Tailor

Non-templated 3'-uridylation of RNAs has emerged as an important mechanism for regulating the processing, stability and biological function of eukaryotic transcripts. In Drosophila, oligouridine tailing by the terminal uridylyl transferase (TUTase) Tailor of numerous RNAs induces their degradation by the exonuclease Dis3L2, which serves functional roles in RNA surveillance and mirtron RNA biogenesis. Tailor preferentially uridylates RNAs terminating in guanosine or uridine nucleotides but the structural basis underpinning its RNA substrate selectivity is unknown. Here, we report crystal structures of Tailor bound to a donor substrate analog or mono- and oligouridylated RNA products. These structures reveal specific amino acid residues involved in donor and acceptor substrate recognition, and complementary biochemical assays confirm the critical role of an active site arginine in conferring selectivity toward 3'-guanosine terminated RNAs. Notably, conservation of these active site features suggests that other eukaryotic TUTases, including mammalian TUT4 and TUT7, might exhibit similar, hitherto unknown, substrate selectivity. Together, these studies provide critical insights into the specificity of 3'-uridylation in eukaryotic post-transcriptional gene regulation

Molecular basis for cytoplasmic RNA surveillance by uridylation-triggered decay in Drosophila

The posttranscriptional addition of nucleotides to the 3' end of RNA regulates the maturation, function, and stability of RNA species in all domains of life. Here, we show that in flies, 3' terminal RNA uridylation triggers the processive, 3'-to-5' exoribonucleolytic decay via the RNase II/R enzyme CG16940, a homolog of the human Perlman syndrome exoribonuclease Dis3l2. Together with the TUTase Tailor, dmDis3l2 forms the cytoplasmic, terminal RNA uridylation-mediated processing (TRUMP) complex that functionally cooperates in the degradation of structured RNA RNA immunoprecipitation and high-throughput sequencing reveals a variety of TRUMP complex substrates, including abundant non-coding RNA, such as 5S rRNA, tRNA, snRNA, snoRNA, and the essential RNase MRP Based on genetic and biochemical evidence, we propose a key function of the TRUMP complex in the cytoplasmic quality control of RNA polymerase III transcripts. Together with high-throughput biochemical characterization of dmDis3l2 and bacterial RNase R, our results imply a conserved molecular function of RNase II/R enzymes as "readers" of destabilizing posttranscriptional marks-uridylation in eukaryotes and adenylation in prokaryotes-that play important roles in RNA surveillance

S129A aSyn is more toxic for yeast cells than the WT aSyn.

<p>(A) Growth curve based on culture OD_600nm of yeast cells expressing WT (□), S129A (▴) or S129E (○) aSyn-GFP, compared to cells that are not expressing the human protein (◊). Cells used as inoculum were exponential-phase cells cultivated in raffinose medium that at time zero were transferred to galactose medium to induce aSyn-GFP expression. (B) aSyn-GFP versus Propidium Iodide (PI) fluorescence and Frequency of PI and GFP positive cells assessed by flow cytometry, in the indicated yeast cells, after 6 hours of aSyn expression induction (***p<0.001; one way ANOVA with Bonferroni's multiple comparison test). A representative result is shown from at least five independent experiments. Values represent the mean ± SD.</p