Disability, fatigue, pain and their associates in early diffuse cutaneous systemic sclerosis: the European Scleroderma Observational Study

Objectives; Our aim was to describe the burden of early dcSSc in terms of disability, fatigue and pain in the European Scleroderma Observational Study cohort, and to explore associated clinical features. Methods; Patients completed questionnaires at study entry, 12 and 24 months, including the HAQ disability index (HAQ-DI), the Cochin Hand Function Scale (CHFS), the Functional Assessment of Chronic Illness Therapy-fatigue and the Short Form 36 (SF36). Associates examined included the modified Rodnan skin score (mRSS), current digital ulcers and internal organ involvement. Correlations between 12-month changes were also examined. Results; The 326 patients recruited (median disease duration 11.9 months) displayed high levels of disability [mean (S.D.) HAQ-DI 1.1 (0.83)], with ‘grip’ and ‘activity’ being most affected. Of the 18 activities assessed in the CHFS, those involving fine finger movements were most affected. High HAQ-DI and CHFS scores were both associated with high mRSS (ρ = 0.34, P < 0.0001 and ρ = 0.35, P < 0.0001, respectively). HAQ-DI was higher in patients with digital ulcers (P = 0.004), pulmonary fibrosis (P = 0.005), cardiac (P = 0.005) and muscle involvement (P = 0.002). As anticipated, HAQ-DI, CHFS, the Functional Assessment of Chronic Illness Therapy and SF36 scores were all highly correlated, in particular the HAQ-DI with the CHFS (ρ = 0.84, P < 0.0001). Worsening HAQ-DI over 12 months was strongly associated with increasing mRSS (ρ = 0.40, P < 0.0001), decreasing hand function (ρ = 0.57, P < 0.0001) and increasing fatigue (ρ = −0.53, P < 0.0001). Conclusion; The European Scleroderma Observational Study highlights the burden of disability in early dcSSc, with high levels of disability and fatigue, associating with the degree of skin thickening (mRSS). Impaired hand function is a major contributor to overall disability

Mechanism of trans-histone crosstalk in the fission yeast «Schizosaccharomyces pombe»

Covalent histone modifications play an important role in the regulation of transcription in all eukaryotic organisms. Methylation of the histone H3 on lysine 4 (H3K4) is an ancient histone mark that is associated with active transcription and serves as a binding site for a number of transcriptional activators. Enzymes that catalyze this modification have been linked to a variety of human pathologies including cancer and mental retardation. The mechanisms that regulate formation of H3K4 methylation during transcription remain poorly understood. H3K4 methylation is intimately connected with ubiquitination of histone H2B (uH2B), another conserved modification found at actively transcribed genes. In all organisms studied, uH2B is a critical activator of H3K4 methylation, and in yeast uH2B is an absolute requirement for processive methylation at this site. The basis for this trans-histone "crosstalk" pathway remains controversial, with some studies suggesting a direct stimulation of methyltransferase activity by uH2B, and others suggesting indirect effects of uH2B on the assembly and stability of methyltransferase complexes.Using unique recombinant chromatin substrates containing semi-synthetic ubiquitinated H2B, I successfully reproduced the trans-histone crosstalk in vitro and showed a direct stimulation of the modified histone H2B on the activity of the native Set1C H3K4 methyltransferase complex purified from the fission yeast Schizosaccharomyces pombe. In my in vitro system, uH2B was an absolute requirement for methyltransferase activity on nucleosomal histone H3, and could also modulate activity on H3 outside the nucleosome, consistent with direct contact between uH2B and the Set1C. Finally, I showed that Set1C purified from strains of S.pombe deficient in H2B ubiquitination had apparently wild-type activity and subunit composition, suggesting that uH2B does not act by altering Set1C stability in this organism. Altogether, my data suggests that H2B ubiquitination can directly stimulate the catalytic activity of the Set1C and that this might represent a fundamental and evolutionary conserved mechanism underlying uH2B/H3K4 methylation crosstalk.Les modifications covalentes des histones jouent un rôle très important dans la régulation de la transcription chez tous les eucaryotes. La méthylation de l'histone 3 sur la lysine 4 (H3K4) est une marque d'origine primitive associée avec une transcription active des gènes qui sert de plateforme d'attachement pour un nombre important d'activateurs de transcription. Les enzymes qui catalysent cette modification sont associées à une grande variété de pathologies humaines telles que le cancer et des formes de retard intellectuel. Les mécanismes qui régularisent la méthylation de H3K4 pendant la transcription ne sont pas bien établis et en général mal compris. La méthylation de H3K4 est intimement reliée à l'ubiquitination de l'histone H2B, une autre modification au niveau des histones très conservée et aussi impliquée dans l'activation de la transcription. Dans tous les organismes étudiés, H2B ubiquitiné (H2Bu) fut désigné comme étant un facteur crucial pour l'activation de la méthylation de H3K4 et constitue chez les levures, une nécessité absolue pour obtenir une réaction de méthylation efficace sur ce site. La mécanistique entourant la diaphonie entre ces histones est controversée; alors que certaines études laissent croire à une stimulation directe de l'activité de méthyltransferase par H2Bu, d'autres sous-entendent un effet indirect employé par H2Bu pour contrôler l'assemblage et la stabilité du complexe de méthyltransferase. En utilisant des substrats uniques de chromatine qui contenaient des histones H2Bu semi-synthétiques, j'ai réussi à reproduire la diaphonie entre H2Bu et H3K4 méthylé in vitro et démontré un effet direct exercé par H2Bu sur la stimulation de l'activité d'un complexe de méthyltransferase natif, Set1C qui fut purifié à partir de la levure de fission Schizosaccharomyces pombe. Dans ce système in vitro, H2Bu était nécessaire pour permettre la méthylation de H3 lorsque ce-dernier était intégré dans un nucléosome. De plus, la présence de H2Bu stimulait l'activité catalytique du complexe Set1C sur des histones H3 hors d'un contexte nucleosomal, ce qui s'accorde avec l'hypothèse d'un contact direct prenant place entre uH2B et Set1C. Finalement, j'ai observé que des complexes Set1C purifiés à partir de variétés de S. pombe déficientes en H2Bu démontraient une activité et une composition comparable à celle de complexes purifiées de variétés sauvages signifiant que l'effet de H2Bu sur Set1C in vivo n'est probablement pas dû à un changement au niveau de la stabilité du complexe dans cet organisme. En conclusion, ces résultats démontrent que l'ubiquitination de H2B peut stimuler l'activité catalytique de Set1C par des moyens directs qui pourraient représenter, d'un point de vue évolutif, un mécanisme fondamental et conservé qui participe à la diaphonie entre H2Bu et H3K4 méthylé

Multiple-target tracking of 3D fluorescent objects based on simulated annealing

International audienceThis paper presents a framework for tracking fluorescent objects imaged with 3D+time video-microscopy. The proposed technique solves the NP-hard problem of the multi-frame object correspondence. For this purpose, we use the simulated annealing algorithm for its flexibility for feature correspondence and its ability to give results in a very short time. Our approach takes into account events like "split", 'merge", "birth" and "death". These "evolutionary events" are motivated by biological and physical considerations. We demonstrate the performance of the proposed algorithm on images of carcinoma NBT-II cells expressing fluorescent LAR-PTP, a tyrosine phosphatase possibly involved in tumor progression

An Ethics Action Plan for Rare Disease Care: Participatory Action Research Approach

BackgroundOwing to their low prevalence, rare diseases are poorly addressed in the scientific literature and clinical practice guidelines. Thus, health care workers are inadequately equipped to provide timely diagnoses, appropriate treatment, and support for these poorly understood conditions. These clinical tribulations are experienced as moral challenges by patients, jeopardizing their life trajectories, dreams, and aspirations. ObjectiveThis paper presents an ethical action plan for rare disease care and the process underlying its development. MethodsThis action plan was designed through an ethical inquiry conducted by the Ethics and Rare Diseases Working Group, which included 3 patient partners, 2 clinician researchers, and 1 representative from Québec’s rare disease association. ResultsThe plan is structured into 4 components. Component A presents the key moral challenges encountered by patients, which are the lack of knowledge on rare diseases among health care workers, the problematic attitudes that it sometimes elicits, and the distress and powerlessness experienced by patients. Component B emphasizes a vision for patient partnership in rare disease care characterized by open-mindedness, empathy, respect, and support of patient autonomy from health care workers. Component C outlines 2 courses of action prompted by this vision: raising awareness among health care workers and empowering patients to better navigate their care. Component D compares several interventions that could help integrate these 2 courses of action in rare disease care. ConclusionsOverall, this action plan represents a toolbox that provides a review of multiple possible interventions for policy makers, hospital managers, practitioners, researchers, and patient associations to critically reflect on key moral challenges experienced by patients with rare diseases and ways to mitigate them. This paper also prompts reflection on the values underlying rare disease care, patient experiences, and health care workers’ beliefs and behaviors. Health care workers and patients were the primary beneficiaries of this action plan

Comment transmettre adéquatement l’information sur l’approvisionnement en médicaments critiques en établissement dans un contexte de pandémie ?

Objectif : Déterminer l’appréciation des équipes relativement aux modalités de diffusion de l’information sur les approvisionnements en médicaments critiques en contexte de pandémie grâce à l’avis des principaux acteurs concernés au Centre hospitalier universitaire de Québec–Université Laval en juillet-août 2020.  Description de la problématique : On dispose de peu de documentation sur les moyens de communication optimaux à utiliser dans un contexte de pandémie. Lors de la première vague de la pandémie de SRAS-CoV-2, un outil de communication a été mis au point et diffusé pour favoriser l’usage optimal des médicaments critiques et ainsi éviter des ruptures d’approvisionnement. Une évaluation de la stratégie de communication utilisée est nécessaire dans l’éventualité de vagues subséquentes.  Résolution de la problématique : Une approche méthodologique mixte a été menée pour ce projet. Un questionnaire en ligne destiné aux pharmaciens, des entretiens individuels ciblés avec des chefs de départements médicaux et un questionnaire soumis par courriel à d’autres centres hospitaliers ont permis de déterminer les différents enjeux permettant de soutenir l’élaboration d’une stratégie de communication efficace en temps de pandémie. Une revue de presse réalisée entre février et juillet 2020 est aussi venue compléter l’analyse.  Conclusion : Les résultats obtenus grâce à cette initiative permettent de déterminer comment et à qui diffuser l’information en contexte de pandémie. Les moyens de communication mis au point par l’établissement ont satisfait les professionnels consultés, et les recommandations formulées permettront de les optimiser pour faire face à de prochaines vagues ou à d’autres événements semblables.  Abstract  Objective: To determine the teams’ assessment of the modalities for disseminating information regarding critical drug supplies during a pandemic, based on the opinions of the main parties concerned at the Centre hospitalier universitaire de Québec-Université Laval in July and August 2020.  Problem description: There is little literature on the optimal means of communication to be used during a pandemic. During the first wave of the SARS-CoV-2 pandemic, a communication tool was developed and disseminated to promote the optimal use of critical drugs in order to prevent supply disruptions. An evaluation of the communication strategy used is needed in the event of subsequent waves.  Problem resolution: A mixed methods approach was used for this project. An online questionnaire for pharmacists, targeted one-on-one interviews with medical department heads and an email questionnaire sent to other hospitals were used to identify the different issues in order to support the development of an effective pandemic communication strategy. A press review conducted between February and July 2020 supplemented the analysis.  Conclusion: The results obtained thanks to this initiative will help determine how and to whom information should be disseminated during a pandemic. The professionals consulted were satisfied with the communication methods developed by the facility, and the recommendations made will enable them to be optimized to deal with future waves or other similar events.

Associations with early-life socio-economic position in adult DNA methylation

Background Disadvantaged socio-economic position (SEP) in childhood is associated with increased adult mortality and morbidity. We aimed to establish whether childhood SEP was associated with differential methylation of adult DNA. Methods Forty adult males from the 1958 British Birth Cohort Study were selected from SEP extremes in both early childhood and mid-adulthood. We performed genome-wide methylation analysis on blood DNA taken at 45 years using MeDIP (methylated DNA immunoprecipitation). We mapped in triplicate the methylation state of promoters of approximately 20 000 genes and 400 microRNAs. Probe methylation scores were averaged across triplicates and differential methylation between groups of individuals was determined. Differentially methylated promoter sites of selected genes were validated using pyrosequencing of bisulfite-converted DNA. Results Variably methylated probes (9112 from n = 223 359 on the microarray) corresponded to 6176 gene promoters with at least one variable probe. Unsupervised hierarchical clustering of probes obtained from the 500 most variable promoters revealed a cluster enriched with high SEP individuals confirming that SEP differences contribute to overall epigenetic variation. Methylation levels for 1252 gene promoters were associated with childhood SEP vs 545 promoters for adulthood SEP. Functionally, associations with childhood SEP appear in promoters of genes enriched in key cell signalling pathways. The differentially methylated promoters associated with SEP cluster in megabase-sized regions of the genome. Conclusions Adult blood DNA methylation profiles show more associations with childhood SEP than adult SEP. Organization of these associations across the genome suggests a well-defined epigenetic pattern linked to early socio-economic environment

A positive feedback loop links opposing functions of P-TEFb/Cdk9 and histone H2B ubiquitylation to regulate transcript elongation in fission yeast.

Transcript elongation by RNA polymerase II (RNAPII) is accompanied by conserved patterns of histone modification. Whereas histone modifications have established roles in transcription initiation, their functions during elongation are not understood. Mono-ubiquitylation of histone H2B (H2Bub1) plays a key role in coordinating co-transcriptional histone modification by promoting site-specific methylation of histone H3. H2Bub1 also regulates gene expression through an unidentified, methylation-independent mechanism. Here we reveal bidirectional communication between H2Bub1 and Cdk9, the ortholog of metazoan positive transcription elongation factor b (P-TEFb), in the fission yeast Schizosaccharomyces pombe. Chemical and classical genetic analyses indicate that lowering Cdk9 activity or preventing phosphorylation of its substrate, the transcription processivity factor Spt5, reduces H2Bub1 in vivo. Conversely, mutations in the H2Bub1 pathway impair Cdk9 recruitment to chromatin and decrease Spt5 phosphorylation. Moreover, an Spt5 phosphorylation-site mutation, combined with deletion of the histone H3 Lys4 methyltransferase Set1, phenocopies morphologic and growth defects due to H2Bub1 loss, suggesting independent, partially redundant roles for Cdk9 and Set1 downstream of H2Bub1. Surprisingly, mutation of the histone H2B ubiquitin-acceptor residue relaxes the Cdk9 activity requirement in vivo, and cdk9 mutations suppress cell-morphology defects in H2Bub1-deficient strains. Genome-wide analyses by chromatin immunoprecipitation also demonstrate opposing effects of Cdk9 and H2Bub1 on distribution of transcribing RNAPII. Therefore, whereas mutual dependence of H2Bub1 and Spt5 phosphorylation indicates positive feedback, mutual suppression by cdk9 and H2Bub1-pathway mutations suggests antagonistic functions that must be kept in balance to regulate elongation. Loss of H2Bub1 disrupts that balance and leads to deranged gene expression and aberrant cell morphologies, revealing a novel function of a conserved, co-transcriptional histone modification