Bipotential mouse embryonic liver (BMEL) cells spontaneously express Pdx1 and Ngn3 but do not undergo further pancreatic differentiation upon Hes1 down-regulation

<p>Abstract</p> <p>Background</p> <p>Liver-to-pancreas conversion offers new possibilities for β-cell engineering for type 1 diabetes therapy. Among conceivable sources of liver cells, we focused on BMEL cells. These untransformed mouse embryonic liver cells have been reproducibly isolated from different inbred mice strains and have the potential to differentiate into hepatocytes and cholangiocytes <it>in vitro </it>and <it>in vivo</it>.</p> <p>Findings</p> <p>Strikingly, we find here that adherent BMEL cells display functional similarities with multipotent pancreatic precursor cells, namely Pdx1 and Ngn3 expression, and further express <it>Hnf6 </it>in floating aggregate culture. Hes1, a direct repressor of Ngn3 and pancreatic endocrine commitment, is expressed in adherent BMEL cells and decreases with time in aggregate culture. However, Hes1 decrease fails to initiate activation of late-stage pancreatic endocrine transcription factors.</p> <p>Conclusion</p> <p>Here we report that BMEL cells present features of pancreatic endocrine progenitor cells. In the field of diabetes research, BMEL cells are of potential interest for the study of inductive signals critical for in vitro β-cell maturation in-liver-to-pancreas conversion.</p

Pdx-1 or Pdx-1-VP16 protein transduction induces β-cell gene expression in liver-stem WB cells

<p>Abstract</p> <p>Background</p> <p>Pancreatic duodenal homeobox-1 (<it>Pdx-1</it>) or <it>Pdx-1-VP16 </it>gene transfer has been shown to induce <it>in vitro </it>rat liver-stem WB cell conversion into pancreatic endocrine precursor cells. High glucose conditions were necessary for further differentiation into functional insulin-producing cells. Pdx-1 has the ability to permeate different cell types due to an inherent protein transduction domain (PTD). In this study, we evaluated liver-to-pancreas conversion of WB cells following Pdx-1 or Pdx-1-VP16 protein transduction.</p> <p>Findings</p> <p>WB cells were grown in high glucose medium containing Pdx-1 or Pdx-1-VP16 recombinant proteins for two weeks. β-like cell commitment was analysed by RT-PCR of pancreatic endocrine genes. We found that WB cells in high glucose culture spontaneously express pancreatic endocrine genes (<it>Pdx-1, Ngn3, Nkx2.2, Kir6.2</it>). Their further differentiation into β-like cells expressing genes related to endocrine pancreas development (<it>Ngn3, NeuroD, Pax4, Nkx2.2, Nkx6.1, Pdx-1</it>) and β-cell function (<it>Glut-2, Kir6.2, insulin</it>) was achieved only in the presence of Pdx-1(-VP16) protein.</p> <p>Conclusion</p> <p>These results demonstrate that Pdx-1(-VP16) protein transduction is instrumental for <it>in vitro </it>liver-to-pancreas conversion and is an alternative to gene therapy for β-cell engineering for diabetes cell therapy.</p

New directions for patient-centred care in scleroderma : the Scleroderma Patient-centred Intervention Network (SPIN)

Systemic sclerosis (SSc), or scleroderma, is a chronic multisystem autoimmune disorder characterised by thickening and fibrosis of the skin and by the involvement of internal organs such as the lungs, kidneys, gastrointestinal tract, and heart. Because there is no cure, feasibly-implemented and easily accessible evidence-based interventions to improve health-related quality of life (HRQoL) are needed. Due to a lack of evidence, however, specific recommendations have not been made regarding non-pharmacological interventions (e.g. behavioural/psychological, educational, physical/occupational therapy) to improve HRQoL in SSc. The Scleroderma Patient-centred Intervention Network (SPIN) was recently organised to address this gap. SPIN is comprised of patient representatives, clinicians, and researchers from Canada, the USA, and Europe. The goal of SPIN, as described in this article, is to develop, test, and disseminate a set of accessible interventions designed to complement standard care in order to improve HRQoL outcomes in SSc.The initial organisational meeting for SPIN was funded by a Canadian Institutes of Health Research (CIHR) Meetings, Planning, and Dissemination grant to B.D. Thombs (KPE-109130), Sclerodermie Quebec, and the Lady Davis Institute for Medical Research of the Jewish General Hospital, Montreal, Quebec. SPIN receives finding support from the Sclemderma Society of Ontario, the Scleroderma Society of Canada, and Sclerodermie Quebec. B.D. Thombs and M. Hudson are supported by New Investigator awards from the CIHR, and Etablissement de Jeunes Chercheurs awards from the Fonds de la Recherche en Sante Quebec (FRSQ). M. Baron is the director of the Canadian Scleroderma Research Group, which receives grant folding from the CIHR, the Scleroderma Society of Canada and its provincial chapters, Scleroderma Society of Ontario, Sclerodermie Quebec, and the Ontario Arthritis Society, and educational grants from Actelion Pharmaceuticals and Pfizer. M.D. Mayes and S. Assassi are supported by the NIH/NIAMS Scleroderma Center of Research Translation grant no. P50-AR054144. S.J. Motivala is supported by an NIH career development grant (K23 AG027860) and the UCLA Cousins Center for Psychoneuroimmunology. D. Khanna is supported by a NIH/NIAMS K23 AR053858-04) and NIH/NIAMS U01 AR057936A, the National Institutes of Health through the NIH Roadmap for Medical Research Grant (AR052177), and has served as a consultant or on speakers bureau for Actelion, BMS, Gilead, Pfizer, and United Therapeutics

The Scleroderma Patient-centered Intervention Network (SPIN) Cohort : protocol for a cohort multiple randomised controlled trial (cmRCT) design to support trials of psychosocial and rehabilitation interventions in a rare disease context

Introduction: Psychosocial and rehabilitation interventions are increasingly used to attenuate disability and improve health-related quality of life (HRQL) in chronic diseases, but are typically not available for patients with rare diseases. Conducting rigorous, adequately powered trials of these interventions for patients with rare diseases is difficult. The Scleroderma Patient-centered Intervention Network (SPIN) is an international collaboration of patient organisations, clinicians and researchers. The aim of SPIN is to develop a research infrastructure to test accessible, low-cost self-guided online interventions to reduce disability and improve HRQL for people living with the rare disease systemic sclerosis (SSc or scleroderma). Once tested, effective interventions will be made accessible through patient organisations partnering with SPIN. Methods and analysis: SPIN will employ the cohort multiple randomised controlled trial (cmRCT) design, in which patients consent to participate in a cohort for ongoing data collection. The aim is to recruit 1500– 2000 patients from centres across the world within a period of 5 years (2013–2018). Eligible participants are persons ≥18 years of age with a diagnosis of SSc. In addition to baseline medical data, participants will complete patient-reported outcome measures every 3 months. Upon enrolment in the cohort, patients will consent to be contacted in the future to participate in intervention research and to allow their data to be used for comparison purposes for interventions tested with other cohort participants. Once nterventions are developed, patients from the cohort will be randomly selected and offered interventions as part of pragmatic RCTs. Outcomes from patients offered interventions will be compared with outcomes from trial-eligible patients who are not offered the interventions. Ethics and dissemination: The use of the cmRCT design, the development of self-guided online interventions and partnerships with patient organisations will allow SPIN to develop, rigourously test and effectively disseminate psychosocial and rehabilitation interventions for people with SSc.(undefined

Revisiting Gender Differences in Somatic Symptoms of Depression: Much Ado about Nothing?

Women have a higher prevalence of Major Depressive Disorder (MDD) and report more severe depressive symptoms than men. Several studies have suggested that gender differences in depression may occur because women report higher levels of somatic symptoms than men. Those studies, however, have not controlled or matched for non-somatic symptoms. The objective of this study was to examine if women report relatively more somatic symptoms than men matched on cognitive/affective symptoms.Male and female patients receiving treatment for MDD in outpatient psychiatric clinics in New Jersey and Pennsylvania, USA were matched on Beck Depression Inventory-II (BDI-II) cognitive/affective symptom scores. Male and female BDI-II somatic symptom scores were compared using independent samples 2-tailed t-tests.Of 472 male and 1,026 female patients, there were 470 male patients (mean age = 40.1 years, SD = 15.1) and 470 female patients (mean age = 43.1 years, SD = 17.2) successfully matched on BDI-II cognitive/affective symptom scores. Somatic symptoms accounted for 35% of total BDI-II scores for male patients versus 38% for matched female patients. Female patients had somatic symptom scores on average 1.3 points higher than males (p<.001), equivalent to 4% of the total BDI-II scores of female patients. Only 5% of male patients and 7% of female patients scored 2 or higher on all BDI-II somatic symptom items.Gender differences in somatic scores were very small. Thus, differences in the experience and reporting of somatic symptoms would not likely explain gender differences in depression rates and symptom severity

Validation of the Body Concealment Scale for Scleroderma (BCSS): Replication in the Scleroderma Patient-centered Intervention Network (SPIN) Cohort

© 2016 Elsevier Ltd Body concealment is an important component of appearance distress for individuals with disfiguring conditions, including scleroderma. The objective was to replicate the validation study of the Body Concealment Scale for Scleroderma (BCSS) among 897 scleroderma patients. The factor structure of the BCSS was evaluated using confirmatory factor analysis and the Multiple-Indicator Multiple-Cause model examined differential item functioning of SWAP items for sex and age. Internal consistency reliability was assessed via Cronbach's alpha. Construct validity was assessed by comparing the BCSS with a measure of body image distress and measures of mental health and pain intensity. Results replicated the original validation study, where a bifactor model provided the best fit. The BCSS demonstrated strong internal consistency reliability and construct validity. Findings further support the BCSS as a valid measure of body concealment in scleroderma and provide new evidence that scores can be compared and combined across sexes and ages

The influence of somatic symptoms in assessing symptoms of depression: do physical health and gender matter?

Major Depressive Disorder (MDD) is common among people with medical illnesses and is more common in women compared to men. In order for comparisons of MDD prevalence rates and self-reported symptom severity across groups to be valid, it is important to establish that diagnostic methods or depression symptom questionnaires that assess severity are measuring the same construct across the groups. There is relatively little research that has attempted to dissect the potential influence of somatic symptom reporting on depression symptom severity ratings, and existing research has important limitations. Thus, the objective of the studies in this thesis was to assess the degree to which differences in somatic symptom reporting might influence scores on a commonly used self-report depression symptom questionnaire, the Beck Depression Inventory (BDI), among post-myocardial infarction (MI) patients, who have a high somatic burden associated with acute cardiovascular disease, compared to psychiatry outpatients, as well as scores on the BDI-II among female compared to male psychiatry outpatients. The first study found evidence that assessing depression symptoms with the BDI post-MI may exaggerate depression symptom severity scores due to the influence of somatic symptoms that commonly occur post-MI, but which are not due to depression. These results, considered in the context of a previous study of the BDI-II that did not find any differences between post-MI and psychiatry outpatients, suggest that the BDI-II may be preferable to the BDI in epidemiological studies of depression among post-MI patients. However, more studies are needed that directly compare the influence of somatic symptoms on the BDI and BDI-II among cardiovascular disease patients. The results of the second study suggest that men and women report similar levels of cognitive/affective and somatic symptoms of depression. Previous suggestions that women report more somatic symptoms of depression than men and that this explains gender differences in depression rates and symptom severity are not supported by the evidence, and future studies should focus on alternative explanations.Le Trouble Dépressif Majeur (TDM) est fréquent chez les personnes malades et plus commun chez les femmes que chez les hommes. Afin d'assurer la validité des comparaisons du taux de prévalence de TDM et des sévérités des symptômes auto-déclarés parmi les groupes, il est important d'établir que les méthodes de diagnostic et les questionnaires sur les symptômes de dépression qui évaluent la gravité mesurent le même concept entre ces groupes. Il y a relativement peu de recherches qui ont tenté de comprendre l'influence potentielle des symptômes somatiques rapportés sur le taux de sévérité des symptômes de dépression, et les études existantes montrent des limitations importantes. Ainsi, l'objectif de la première étude de cette thèse était d'évaluer à quel point les symptômes somatiques rapportés pouvaient influencer les résultats sur un questionnaire des symptômes auto-déclarés de dépression (le "Beck Depression Inventory (BDI)") parmi les patients qui ont subi un infarctus du myocarde (IM) et qui ont des limites physiques importantes associées à cette maladie, comparés aux patients externe de psychiatrie. L'objectif de la deuxième études était aussi d'évaluer à quel point les symptômes somatiques rapportés pouvaient influencer les résultats sur un autre questionnaire des symptômes auto-déclarés de dépression (le "BDI-II") parmi les patientes externes femelles en psychiatrie par rapport aux patients externes mâles. La première étude a démontré que l'évaluation des symptômes de dépression avec le BDI parmi les patients qui ont subi un IM pouvait exagérer la gravité des symptômes dus à l'influence des symptômes somatiques qui surviennent fréquemment suite à un IM, mais qui ne seraient pas dus à la dépression. Ces résultats, considérés dans le contexte d'une étude précédente du BDI-II qui n'a pas identifié de différences entre les patients qui ont subi un IM et les patients externes de psychiatrie, suggère que le BDI-II peut-être préférable au BDI dans les études épidémiologiques de la dépression chez les patients qui ont subi un IM. Cependant, d'autres études sont nécessaires pour comparer directement l'influence des symptômes somatiques sur le BDI et le BDI-II chez les patients souffrant de maladies cardiovasculaires. Les résultats de la deuxième étude suggèrent que les hommes et les femmes présentent des niveaux similaires de symptômes cognitifs/affectifs et somatiques de dépression. Les résultats de cette deuxième étude ne supportent pas les conclusions des études précédentes qui suggèrent que les femmes rapportent plus de symptômes somatiques de dépression que les hommes et que cela expliquerait les différences entre les sexes dans les taux de dépression et la sévérité des symptômes. Les études futures devraient se concentrer sur d'autres explications

Support groups in a rare disease context: exploring the training needs of scleroderma support group facilitators and factors affecting participation in scleroderma support groups

The growing number of Canadians with one or more chronic illnesses, combined with increasingly limited healthcare resources, has made it difficult for healthcare providers and patient organizations to provide the support that people with chronic diseases need in order to deal with their conditions. As such, many patients turn to support groups to help them cope with and manage their disease. In common diseases, support groups are often facilitated by professionals, but in rare diseases, where barriers to providing support are even more pronounced, support groups are typically led by other patients. Scleroderma (or systemic sclerosis, SSc) is an example of a rare disease where peer facilitated support groups play an important role for people living with the disease. The Scleroderma Society of Canada and the Scleroderma Foundation in the United States are committed to increasing the accessibility and effectiveness of SSc support groups. To do this, they have partnered with our research team to develop an educational training program for peer facilitators of SSc support groups. In order to inform the development of this program four independent studies were conducted and included in this thesis. Study 1 was a systematic review examining the existence of training programs for peer facilitators of illness-based support groups and the effect of such programs on 1) the competency and self-efficacy of group facilitators and 2) self-efficacy for disease management, health outcomes, and satisfaction with the support group experience among group members. The findings of the review highlighted the need for training programs for peer facilitators of illness-based support groups and for well-designed and executed trials that evaluate whether these programs improve outcomes among group facilitators and members. Study 2 was a scoping review exploring the 1) benefits of participating in rare disease support groups and 2) facilitators and barriers of initiating and sustaining these groups. The findings of the review suggested that support groups are an important resource for people with rare diseases and that training programs for peer facilitators of rare disease support groups, such as SSc support groups, could be useful in addressing challenges regarding establishing and maintaining these groups. Studies 3 and 4 employed self-report measures to identify 1) the training and support needs of SSc support group facilitators and 2) the reasons people with SSc do not participate in SSc support groups. These findings underscored the importance of an educational training program for peer facilitators of SSc support group and highlighted areas of focus for this program. Implications and future directions for research are discussed.Il est de plus en plus difficile pour les professionnels des soins de la santé ainsi qu'aux organismes de patients, de fournir les soins requis aux gens atteints de maladies chroniques principalement à cause du nombre croissant de canadiens ayant une ou plusieurs de ces maladies, et des ressources de plus en plus limitées dédiées aux soins de la santé. Par conséquent, plusieurs patients se tournent vers des groupes de soutien pour les aider à affronter et gérer leur maladie. Dans les cas de maladies plus communes, les groupes de soutien des patients sont souvent gérés par des professionnels alors que dans le cas de maladie plus rares, où les obstacles pour fournir de tels services sont plus importants, les groupes de soutien sont typiquement gérés par d'autres patients. La sclérodermie (sclérose systémique, ScS) est un exemple de maladie rare où un groupe de soutien guidé par un pair joue un rôle important pour les gens vivant avec cette maladie. La Société de sclérodermie du Canada et la fondation américaine de sclérodermie se sont engagés à augmenter l'accessibilité et l'efficacité des groupes de soutien ScS. Pour ce faire, ils ont fait un partenariat avec notre équipe de recherche pour développer un programme de formation destiné aux leaders de ces groupes de soutien ScS. Dans le but de supporter le développement de ce programme, quatre études indépendantes ont été réalisées et sont incluses dans cette thèse. L'étude 1 est une revue systématique portant sur l'examen des programmes de formation existants pour les leaders des groupes de soutien et les impacts de tels programmes sur 1) la compétence et l'efficacité des leaders 2) l'efficacité de la gestion de la maladie, les résultats sur la santé et la satisfaction des membres de ces groupes de soutien. Les conclusions de cette première étude ont mis en lumière le besoin de programmes de formation pour les leaders des groupes de soutien, ainsi que la conception minutieuse et l'exécution de tests afin d'évaluer si ces programmes améliorent la performance des leaders et le bien-être des membres. L'étude 2 est une revue de la portée visant 1) les bénéfices reliés à la participation à des groupes de soutien aux maladies rares, et 2) les facilitateurs et les obstacles reliés à la création et au maintient de ces groupes. Les conclusions de cette deuxième étude indiquent que les groupes de support sont une ressource importante pour les gens avec des maladies rares et que les programmes de formation des leaders des groupes de soutien au maladies rares, tel que le groupe de soutien ScS, pourrait être utile pour confronter les défis reliés à la mise en place et au soutien de ces groupes. Les études 3 et 4 ont employés des méthodes d'auto-évaluation afin d'identifier 1) la formation et les besoins de support des leaders des groupes de soutien ScS et 2) les raisons pour lesquelles les gens avec ScS ne participent pas à des groupes de soutien. Les résultats des ces études soulignent l'importance des programmes de formation pour les leaders des groupes de soutien ScS et mets en lumière les principaux domaines ciblés par ces programmes de formation. Les implications ainsi que des directions futures pour la recherche sont aussi discutées

Étude de la différenciation de cellules souches hépatiques en cellules productrices d insuline (effets de la transduction des protéines Pdx-1 ou Pdx-1-VP16 dans les lignées cellulaires WB et BMEL )

Une approche thérapeutique du diabète de type I consiste à développer des sources alternatives de cellules ß afin d en disposer en quantité non limitante en vue de leur transplantation. Du fait de l origine endodermique et de propriétés physiologiques communes avec les cellules ß, les cellules hépatiques sont des bonnes candidates. Dans ces cellules, la surexpression du gène codant pour Pdx-1, facteur clé du développement pancréatique et de la fonction ß, induit leur conversion en cellules productrices d insuline. La fusion de Pdx-1 à l activateur viral VP16 renforce cette action. Par ailleurs, la protéine Pdx-1 contient un peptide de transduction (PTD) lui permettant de traverser les membranes. L objectif de cette étude a été d évaluer les effets de la transduction des protéines Pdx-1 ou Pdx-1-VP16 sur la différenciation de deux lignées de cellules souches hépatiques, WB et BMEL, en cellules productrices d insuline. Nous avons produit les protéines Pdx-1(-VP16) et vérifié leur capacité de transduction ainsi que leur fonctionnalité. Puis, nous avons montré que les protéines Pdx-1(-VP16) couplées à un milieu riche en glucose engagent les cellules WB, mais pas les cellules BMEL, vers un phénotype pancréatique endocrine ß. Une autre approche consistant à inhiber l expression de Hes1, un facteur limitant la différenciation pancréatique endocrine exprimé par les BMEL, n a pas permis d orienter ces cellules vers un phénotype ß. Le traitement par la protéine Pdx-1 ou Pdx-1-VP16 pourrait donc constituer, sous certaines conditions, un nouvel outil pour générer des cellules productrices d insuline à partir de cellules hépatiques ainsi qu une alternative à la thérapie géniqueA new approach of type I diabetes therapy consists in developing alternative and renewable sources of transplantable ß cells. As liver and pancreas share endodermal origin and physiological properties, hepatic cells seem to be suitable candidates. Overexpression of the gene coding for Pdx-1, a key factor of pancreatic development and ß cell function, induces conversion of hepatic cells into insulin-producing cells. Fusion of Pdx-1 to the potent viral activator VP16 enhances this action. Furthermore, Pdx-1 contains a Protein Transduction Domain (PTD) enabling it to cross membranes. The aim of our study was to evaluate effects of Pdx-1 or Pdx-1-VP16 protein transduction on liver-to-ß-like cell conversion in two hepatic stem cell lines i.e. WB and BMEL cells. First, we produced Pdx-1(-VP16) recombinant proteins and checked their transduction and functionality. Then we proved that Pdx-1(-VP16) protein transduction in conjunction with high glucose culture induces ß-like cell commitment in WB cells, but not in BMEL cells. Repression of Hes1, a factor expressed in BMEL cells which hampers endocrine specification, did not direct these cells towards a pancreatic fate. In particular conditions, Pdx-1 or Pdx-1-VP16 protein treatment may be instrumental for in vitro liver-to-pancreas conversion and represents an alternative to gene therapy for ß cell engineering for cell therapy of diabetes.NANTES-BU Sciences (441092104) / SudocSudocFranceF

An Assessment of the Measurement Equivalence of English and French Versions of the Center for Epidemiologic Studies Depression (CES-D) Scale in Systemic Sclerosis

<div><p>Objectives</p><p>Center for Epidemiologic Studies Depression (CES-D) Scale scores in English- and French-speaking Canadian systemic sclerosis (SSc) patients are commonly pooled in analyses, but no studies have evaluated the metric equivalence of the English and French CES-D. The study objective was to examine the metric equivalence of the CES-D in English- and French-speaking SSc patients.</p><p>Methods</p><p>The CES-D was completed by 1007 English-speaking and 248 French-speaking patients from the Canadian Scleroderma Research Group Registry. Confirmatory factor analysis (CFA) was used to assess the factor structure in both samples. The Multiple-Indicator Multiple-Cause (MIMIC) model was utilized to assess differential item functioning (DIF).</p><p>Results</p><p>A two-factor model (Positive and Negative affect) showed excellent fit in both samples. Statistically significant, but small-magnitude, DIF was found for 3 of 20 CES-D items, including items 3 (<i>Blues</i>), 10 (<i>Fearful</i>), and 11 (<i>Sleep</i>). Prior to accounting for DIF, French-speaking patients had 0.08 of a standard deviation (SD) lower latent scores for the Positive factor (95% confidence interval [CI]−0.25 to 0.08) and 0.09 SD higher scores (95% CI−0.07 to 0.24) for the Negative factor than English-speaking patients. After DIF correction, there was no change on the Positive factor and a non-significant increase of 0.04 SD on the Negative factor for French-speaking patients (difference = 0.13 SD, 95% CI−0.03 to 0.28).</p><p>Conclusions</p><p>The English and French versions of the CES-D, despite minor DIF on several items, are substantively equivalent and can be used in studies that combine data from English- and French-speaking Canadian SSc patients.</p></div