74 results on '"Jane Sun"'
Search Results
2. A randomized controlled trial evaluating the effects of motivational interviewing in new hearing aid users (MI-HAT): study protocol for a randomized controlled trial
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Alice Q. Liu, Printha Wijesinghe, Melissa Lee, Carol Lau, Jane Sun, and Desmond A Nunez
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Motivational interviewing ,Hearing aid ,Hearing loss ,Randomized controlled trial ,Study protocol ,Medicine (General) ,R5-920 - Abstract
Abstract Background Hearing loss is the third leading global cause of disability and is associated with poorer quality of life. Hearing aids are often recommended for hearing loss; however, hearing aid uptake and use rates are perpetually low. Motivational interviewing (MI) is a patient-centered counseling aimed at addressing the desire in the patient to change their behavior. The aim of this study is to investigate the impact of one-on-one MI sessions on hearing aid use among new adult users. Methods A multi-center, prospective, randomized patient-blind controlled trial with a pre- and post-tests design. New hearing aid users ≥ 18 years of age will be recruited from Vancouver, Canada. They will be randomly assigned to a treatment or control group. The treatment group will attend a one-on-one MI session hosted by a practicing MI therapist in addition to standard in-person audiological care. The control group will receive standard in-person audiological care. Data is collected at baseline and at 1, 3, 6, and 12 months’ follow-ups. The primary outcomes are data-logged hearing aid use hours and patient-reported outcomes as measured by the International Outcome Inventory for Hearing Aids questionnaire. Associations between intervention and hearing aid use hours and self-reported outcome measures will be assessed. Discussion This trial is designed to evaluate the efficacy of one-on-one MI in improving hearing aid use in new adult users in the short and long terms. Results will contribute to the evidence on whether MI counseling has an effect on hearing aid use and may guide future clinical practices. Trial registration ClinicalTrials.gov NCT04673565 . Registered on 17 December 2020.
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- 2023
- Full Text
- View/download PDF
3. Combination of human endothelial colony-forming cells and mesenchymal stromal cells exert neuroprotective effects in the growth-restricted newborn
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Kirat K. Chand, Jatin Patel, S. T. Bjorkman, Seen-Ling Sim, Stephanie M. Miller, Elliot Teo, Lara Jones, Jane Sun, Paul B. Colditz, Kiarash Khosrotehrani, and Julie A. Wixey
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Medicine - Abstract
Abstract The foetal brain is particularly vulnerable to the detrimental effects of foetal growth restriction (FGR) with subsequent abnormal neurodevelopment being common. There are no current treatments to protect the FGR newborn from lifelong neurological disorders. This study examines whether pure foetal mesenchymal stromal cells (MSC) and endothelial colony-forming cells (ECFC) from the human term placenta are neuroprotective through modulating neuroinflammation and supporting the brain vasculature. We determined that one dose of combined MSC-ECFCs (cECFC; 106 ECFC 106 MSC) on the first day of life to the newborn FGR piglet improved damaged vasculature, restored the neurovascular unit, reduced brain inflammation and improved adverse neuronal and white matter changes present in the FGR newborn piglet brain. These findings could not be reproduced using MSCs alone. These results demonstrate cECFC treatment exerts beneficial effects on multiple cellular components in the FGR brain and may act as a neuroprotectant.
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- 2021
- Full Text
- View/download PDF
4. Eliminating the routine use of examination table paper in outpatient oncology clinics.
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Mariano, Caroline, Wells, Hannah, Brown, Maura, Clement, Krista, Wooffindin, Rae, Hare, Kevin, Lefresne, Shilo, Kaur, Jagbir, Darud, Michael, Chui, Vincent, and Jane Sun
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CARBON dioxide mitigation ,GREENHOUSE gases ,CARBON emissions ,MICROBIAL contamination ,INFECTION control - Abstract
Background: Health care systems contribute significantly to greenhouse gas emissions. One source of these emissions is single-use products. Examination table paper does not confer protection against microbial contamination and thus can be omitted while following infection control standards. The objective is to eliminate the routine use of examination table paper in outpatient oncology clinics at BC Cancer. Methods: A quality improvement approach was used. Examination tables continued to be disinfected using wipes between patients, but table paper was not used. Plan-do-studyact cycles were performed at four regional cancer centres. Results: Pre-intervention, the cancer centres used 19 to 69 rolls of paper monthly. Postintervention, usage declined to 0 to 2 rolls monthly. This was associated with annual cost savings of $3974 and a reduction of 32 501 kg of carbon dioxide emissions. Conclusions: The use of examination table paper can be eliminated in outpatient clinics, resulting in both cost savings and a reduction in carbon dioxide emissions. [ABSTRACT FROM AUTHOR]
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- 2024
5. Combination of human endothelial colony-forming cells and mesenchymal stromal cells exert neuroprotective effects in the growth-restricted newborn
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Julie A. Wixey, S. T. Bjorkman, Lara Jones, Seen-Ling Sim, Jatin Patel, Paul B. Colditz, Jane Sun, Kirat K. Chand, Elliot Teo, Kiarash Khosrotehrani, and Stephanie M. Miller
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Day of life ,Biomedical Engineering ,Medicine (miscellaneous) ,Inflammation ,Foetal brain ,Stem cells ,Neuroprotection ,Article ,Term placenta ,03 medical and health sciences ,0302 clinical medicine ,Foetal growth ,Medicine ,Neuroinflammation ,030304 developmental biology ,0303 health sciences ,business.industry ,Mesenchymal stem cell ,Development of the nervous system ,Cell Biology ,3. Good health ,Cancer research ,medicine.symptom ,business ,030217 neurology & neurosurgery ,Developmental Biology - Abstract
The foetal brain is particularly vulnerable to the detrimental effects of foetal growth restriction (FGR) with subsequent abnormal neurodevelopment being common. There are no current treatments to protect the FGR newborn from lifelong neurological disorders. This study examines whether pure foetal mesenchymal stromal cells (MSC) and endothelial colony-forming cells (ECFC) from the human term placenta are neuroprotective through modulating neuroinflammation and supporting the brain vasculature. We determined that one dose of combined MSC-ECFCs (cECFC; 106 ECFC 106 MSC) on the first day of life to the newborn FGR piglet improved damaged vasculature, restored the neurovascular unit, reduced brain inflammation and improved adverse neuronal and white matter changes present in the FGR newborn piglet brain. These findings could not be reproduced using MSCs alone. These results demonstrate cECFC treatment exerts beneficial effects on multiple cellular components in the FGR brain and may act as a neuroprotectant.
- Published
- 2021
6. A randomized controlled trial evaluating the effects of motivational interviewing in new hearing aid users (MI-HAT): study protocol for a randomized controlled trial
- Author
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Alice Q. Liu, Printha Wijesinghe, Melissa Lee, Jane Sun, Carol Lau, and Desmond A. Nunez
- Subjects
Medicine (miscellaneous) ,Pharmacology (medical) - Abstract
Background Hearing loss is the third leading global cause of disability and is associated with poorer quality of life. Hearing aids are often recommended for hearing loss; however, hearing aid uptake and use rates are perpetually low. Motivational interviewing (MI) is a patient-centered counseling aimed at addressing the desire in the patient to change their behavior. The aim of this study is to investigate the impact of one-on-one MI sessions on hearing aid use among new adult users. Methods A multi-center, prospective, randomized patient-blind controlled trial with a pre- and post-tests design. New hearing aid users ≥ 18 years of age will be recruited from Vancouver, Canada. They will be randomly assigned to a treatment or control group. The treatment group will attend a one-on-one MI session hosted by a practicing MI therapist in addition to standard in-person audiological care. The control group will receive standard in-person audiological care. Data is collected at baseline and at 1, 3, 6, and 12 months’ follow-ups. The primary outcomes are data-logged hearing aid use hours and patient-reported outcomes as measured by the International Outcome Inventory for Hearing Aids questionnaire. Associations between intervention and hearing aid use hours and self-reported outcome measures will be assessed. Discussion This trial is designed to evaluate the efficacy of one-on-one MI in improving hearing aid use in new adult users in the short and long terms. Results will contribute to the evidence on whether MI counseling has an effect on hearing aid use and may guide future clinical practices. Trial registration ClinicalTrials.gov NCT04673565. Registered on 17 December 2020.
- Published
- 2022
7. Abstract 188: Autologous organoids and T cell co-cultures as a powerful personalized platform for immunotherapy development
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Soura Mardpour, Claudia Beaurivage, Jane Sun, Lorenz Jahn, Rene Overmeer, Lars-Eric Fielmich, Pleun Hombrink, Farzin Pourfarzad, and Sylvia F. Boj
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Cancer Research ,Oncology - Abstract
Recent advances in cancer immunotherapy had a positive impact on the life expectancy of patients for a large range of clinical indications. With new treatment strategies and druggable targets being identified at an increasing pace, the number of patients eligible for cancer immunotherapy is expected to expand steadily. However, promising therapeutic developments face hurdles in translating preclinical findings into therapy since conventional 2D cancer models hold low clinical predictive value. We developed an innovative alternative, built on the discovery that adult stem cells proliferate and organise into three-dimensional organotypic structures when they are embedded into extracellular matrix. Patient specific organoids are generated from healthy and malignant tissues and stored as biobanks with high quality and reproducibility. HUB Organoids recapitulate complex characteristics of the original parental tissue, including molecular heterogeneity, and morphological and functional traits. Since cancer progression and responses to immunotherapy are governed by immune cell interactions in the tumor microenvironment, we developed an assay in which colorectal (CRC) and non-small cell lung cancer (NSCLC) tumor organoids are co-cultured with simultaneously expanded paired tumor infiltrating lymphocytes (TIL) as a source of tumor reactive cells. Our system allows robust expansion of TIL with preserved T-cell receptor repertoires and has an establishment rate of 75%. Paired resources allow for screening of immune-modulatory therapies under physiologic conditions not depending on peptide-pulsing and allo-reactivity. For this purpose, we developed a organoid-TIL co-culture system which quantifies and characterizes tumor organoid apoptosis by combining image-based analysis with flow cytometry and cytokine release assays as measurements of T-cell cytotoxicity against tumor (and normal-matched) organoids. Our system offers a powerful tool for the development and validation of cancer immunotherapy and may help to stratify cancer patients for susceptibility of various types of immunotherapies. Citation Format: Soura Mardpour, Claudia Beaurivage, Jane Sun, Lorenz Jahn, Rene Overmeer, Lars-Eric Fielmich, Pleun Hombrink, Farzin Pourfarzad, Sylvia F. Boj. Autologous organoids and T cell co-cultures as a powerful personalized platform for immunotherapy development [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2023; Part 1 (Regular and Invited Abstracts); 2023 Apr 14-19; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2023;83(7_Suppl):Abstract nr 188.
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- 2023
8. Surface Degradation Analysis of Commercial Nickel-rich Oxide Cathode Materials by Multiple Electron Microscopy Technologies
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Christopher Kurtz, Jiangtao Zhu, Cynthia Wang, Zhihuan Wang, Jiajie Qian, Shaojie Wang, Yanxiao Ma, Charlene Sun, Tina Hsu, Jane Sun, Xiuhong Han, and Xinyu Zhao
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Nickel ,Materials science ,Chemical engineering ,chemistry ,law ,Degradation (geology) ,chemistry.chemical_element ,Electron microscope ,Instrumentation ,Oxide cathode ,law.invention - Published
- 2021
9. A case of bullous fixed drug eruption caused by tadalafil
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Zhang, Jane Sun, Aggarwal, Ishita, and Bain, Michelle
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- 2024
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10. Evaluation of variability in human kidney organoids
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Ernst J. Wolvetang, Alicia Oshlack, Belinda Phipson, Minoru Takasato, Lorna J Hale, Jane Sun, Hsan Jan Yen, Melissa H. Little, Alexander N. Combes, Luke Zappia, Kynan T. Lawlor, Sara E. Howden, Thomas A. Forbes, and Pei Xuan Er
- Subjects
Transcription, Genetic ,Cellular differentiation ,Induced Pluripotent Stem Cells ,Nephron ,Biology ,Kidney ,Models, Biological ,Biochemistry ,Article ,03 medical and health sciences ,Single-cell analysis ,Organoid ,medicine ,Humans ,Induced pluripotent stem cell ,Molecular Biology ,030304 developmental biology ,0303 health sciences ,Gene Expression Profiling ,Reproducibility of Results ,Kidney metabolism ,Cell Differentiation ,Epithelial Cells ,Cell Biology ,Clone Cells ,Cell biology ,Organoids ,Gene expression profiling ,medicine.anatomical_structure ,Kidney Diseases ,Single-Cell Analysis ,Functional genomics ,Biotechnology - Abstract
The utility of human pluripotent stem cell–derived kidney organoids relies implicitly on the robustness and transferability of the protocol. Here we analyze the sources of transcriptional variation in a specific kidney organoid protocol. Although individual organoids within a differentiation batch showed strong transcriptional correlation, we noted significant variation between experimental batches, particularly in genes associated with temporal maturation. Single-cell profiling revealed shifts in nephron patterning and proportions of component cells. Distinct induced pluripotent stem cell clones showed congruent transcriptional programs, with interexperimental and interclonal variation also strongly associated with nephron patterning. Epithelial cells isolated from organoids aligned with total organoids at the same day of differentiation, again implicating relative maturation as a confounder. This understanding of experimental variation facilitated an optimized analysis of organoid-based disease modeling, thereby increasing the utility of kidney organoids for personalized medicine and functional genomics.
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- 2018
11. Human primed endothelial colony forming cells exert neuroprotective effects in the growth restricted newborn piglet
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Jatin Patel, Lara Jones, Kiarash Khosrotehrani, Tracey Bjorkman, Julie A. Wixey, Stephanie M. Miller, Seen-Ling Sim, Paul B. Colditz, Jane Sun, Kirat K. Chand, and Elliot Teo
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medicine.medical_specialty ,Fetus ,Brain vasculature ,business.industry ,Day of life ,Mesenchymal stem cell ,Inflammation ,Neuroprotection ,Endocrinology ,Internal medicine ,medicine ,medicine.symptom ,Stem cell ,business ,Neuroinflammation - Abstract
The fetal brain is particularly vulnerable to the detrimental effects of fetal growth restriction (FGR) with subsequent abnormal neurodevelopment being common. There are no current treatments to protect the FGR newborn from lifelong neurological disorders. This study examines whether pure fetal mesenchymal stem cells and endothelial colony forming cells (ECFC) from the human term placenta are neuroprotective through modulating neuroinflammation and supporting the brain vasculature. We determined that one dose of these primed ECFCs (pECFC) on the first day of life to the newborn FGR piglet improved damaged vasculature, restored the neurovascular unit, reduced brain inflammation and improved adverse neuronal and white matter changes present in the FGR newborn piglet brain. These findings could not be reproduced using mesenchymal stromal cells alone. These results demonstrate pECFC treatment exerts beneficial effects on multiple cellular components in the FGR brain and act as a neuroprotectant.One Sentence SummaryStem cell treatment improves brain outcomes in the growth restricted newborn
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- 2021
12. Roadmap consensus on carotid artery plaque imaging and impact on therapy strategies and guidelines
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L Saba, W Brinjikji, JP (Jeremiah) Spence, M Wintermark, M Castillo, J.G.G. (Gerard) Borst, Q (Qin) Yang, C Yuan, A. Buckler, M. Edjlali, T Saam, D Saloner, B. K. Lal, D Capodanno, J (Jane) Sun, N. Balu, R Naylor, A. (Aad) van der Lugt, B. A. Wasserman, M. Eline Kooi, J Wardlaw, JH Gillard, Giuseppe Lanzino, U Hedin, D. Mikulis, A (Alok) Gupta, J. K. DeMarco, CP Hess, J Van Goethem, TS Hatsukami, P Rothwell, M.D. (Michael) Brown, A. R. Moody, L Saba, W Brinjikji, JP (Jeremiah) Spence, M Wintermark, M Castillo, J.G.G. (Gerard) Borst, Q (Qin) Yang, C Yuan, A. Buckler, M. Edjlali, T Saam, D Saloner, B. K. Lal, D Capodanno, J (Jane) Sun, N. Balu, R Naylor, A. (Aad) van der Lugt, B. A. Wasserman, M. Eline Kooi, J Wardlaw, JH Gillard, Giuseppe Lanzino, U Hedin, D. Mikulis, A (Alok) Gupta, J. K. DeMarco, CP Hess, J Van Goethem, TS Hatsukami, P Rothwell, M.D. (Michael) Brown, and A. R. Moody
- Abstract
Current guidelines for primary and secondary prevention of stroke in patients with carotid atherosclerosis are based on the quantification of the degree of stenosis and symptom status. Recent publications have demonstrated that plaque morphology and composition, independent of the degree of stenosis, are important in the risk stratification of carotid atherosclerotic disease. This finding raises the question as to whether current guidelines are adequate or if they should be updated with new evidence, including imaging for plaque phenotyping, risk stratification, and clinical decision-making in addition to the degree of stenosis. To further this discussion, this roadmap consensus article defines the limits of luminal imaging and highlights the current evidence supporting the role of plaque imaging. Furthermore, we identify gaps in current knowledge and suggest steps to generate high-quality evidence, to add relevant information to guidelines currently based on the quantification of stenosis.
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- 2021
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13. TWIST1 Homodimers and Heterodimers Orchestrate Lineage-Specific Differentiation
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David A.F. Loebel, Mark E. Graham, Nicolas Fossat, Madeleine Demuth, Patrick P.L. Tam, Xiaochen Fan, Pierre Osteil, Jane Sun, and Ashley J. Waardenberg
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Mesoderm ,Cell type ,Lineage (genetic) ,animal structures ,Epithelial-Mesenchymal Transition ,Mesenchyme ,Biology ,Cell Line ,Madin Darby Canine Kidney Cells ,03 medical and health sciences ,0302 clinical medicine ,Dogs ,medicine ,Animals ,Humans ,Molecular Biology ,030304 developmental biology ,0303 health sciences ,Twist-Related Protein 1 ,Neural crest ,Gene Expression Regulation, Developmental ,Nuclear Proteins ,Cell Differentiation ,Cell Biology ,Embryonic stem cell ,Cell biology ,Mice, Inbred C57BL ,medicine.anatomical_structure ,Neural Crest ,030220 oncology & carcinogenesis ,Mutation ,Stem cell ,Endoderm ,Protein Multimerization ,Transcriptome ,Research Article - Abstract
The extensive array of basic helix-loop-helix (bHLH) transcription factors and their combinations as dimers underpin the diversity of molecular function required for cell type specification during embryogenesis. The bHLH factor TWIST1 plays pleiotropic roles during development. However, which combinations of TWIST1 dimers are involved and what impact each dimer imposes on the gene regulation network controlled by TWIST1 remain elusive. In this work, proteomic profiling of human TWIST1-expressing cell lines and transcriptome analysis of mouse cranial mesenchyme have revealed that TWIST1 homodimers and heterodimers with TCF3, TCF4, and TCF12 E-proteins are the predominant dimer combinations. Disease-causing mutations in TWIST1 can impact dimer formation or shift the balance of different types of TWIST1 dimers in the cell, which may underpin the defective differentiation of the craniofacial mesenchyme. Functional analyses of the loss and gain of TWIST1-E-protein dimer activity have revealed previously unappreciated roles in guiding lineage differentiation of embryonic stem cells: TWIST1-E-protein heterodimers activate the differentiation of mesoderm and neural crest cells, which is accompanied by the epithelial-to-mesenchymal transition. At the same time, TWIST1 homodimers maintain the stem cells in a progenitor state and block entry to the endoderm lineage.
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- 2019
14. Patient-reported outcome measures for monitoring primary care patients with depression: the PROMDEP cluster RCT and economic evaluation
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Tony Kendrick, Christopher Dowrick, Glyn Lewis, Michael Moore, Geraldine M Leydon, Adam WA Geraghty, Gareth Griffiths, Shihua Zhu, Guiqing Lily Yao, Carl May, Mark Gabbay, Rachel Dewar-Haggart, Samantha Williams, Lien Bui, Natalie Thompson, Lauren Bridewell, Emilia Trapasso, Tasneem Patel, Molly McCarthy, Naila Khan, Helen Page, Emma Corcoran, Jane Sungmin Hahn, Molly Bird, Mekeda X Logan, Brian Chi Fung Ching, Riya Tiwari, Anna Hunt, and Beth Stuart
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primary health care ,mental health ,mood disorders ,depression ,patient-reported outcome measures ,cost–benefit analysis ,Medical technology ,R855-855.5 - Abstract
Background Guidelines on the management of depression recommend that practitioners use patient-reported outcome measures for the follow-up monitoring of symptoms, but there is a lack of evidence of benefit in terms of patient outcomes. Objective To test using the Patient Health Questionnaire-9 questionnaire as a patient-reported outcome measure for monitoring depression, training practitioners in interpreting scores and giving patients feedback. Design Parallel-group, cluster-randomised superiority trial; 1 : 1 allocation to intervention and control. Setting UK primary care (141 group general practices in England and Wales). Inclusion criteria Patients aged ≥ 18 years with a new episode of depressive disorder or symptoms, recruited mainly through medical record searches, plus opportunistically in consultations. Exclusions Current depression treatment, dementia, psychosis, substance misuse and risk of suicide. Intervention Administration of the Patient Health Questionnaire-9 questionnaire with patient feedback soon after diagnosis, and at follow-up 10–35 days later, compared with usual care. Primary outcome Beck Depression Inventory, 2nd edition, symptom scores at 12 weeks. Secondary outcomes Beck Depression Inventory, 2nd edition, scores at 26 weeks; antidepressant drug treatment and mental health service contacts; social functioning (Work and Social Adjustment Scale) and quality of life (EuroQol 5-Dimension, five-level) at 12 and 26 weeks; service use over 26 weeks to calculate NHS costs; patient satisfaction at 26 weeks (Medical Informant Satisfaction Scale); and adverse events. Sample size The original target sample of 676 patients recruited was reduced to 554 due to finding a significant correlation between baseline and follow-up values for the primary outcome measure. Randomisation Remote computerised randomisation with minimisation by recruiting university, small/large practice and urban/rural location. Blinding Blinding of participants was impossible given the open cluster design, but self-report outcome measures prevented observer bias. Analysis was blind to allocation. Analysis Linear mixed models were used, adjusted for baseline depression, baseline anxiety, sociodemographic factors, and clustering including practice as random effect. Quality of life and costs were analysed over 26 weeks. Qualitative interviews Practitioner and patient interviews were conducted to reflect on trial processes and use of the Patient Health Questionnaire-9 using the Normalization Process Theory framework. Results Three hundred and two patients were recruited in intervention arm practices and 227 patients were recruited in control practices. Primary outcome data were collected for 252 (83.4%) and 195 (85.9%), respectively. No significant difference in Beck Depression Inventory, 2nd edition, score was found at 12 weeks (adjusted mean difference –0.46, 95% confidence interval –2.16 to 1.26). Nor were significant differences found in Beck Depression Inventory, 2nd Edition, score at 26 weeks, social functioning, patient satisfaction or adverse events. EuroQol-5 Dimensions, five-level version, quality-of-life scores favoured the intervention arm at 26 weeks (adjusted mean difference 0.053, 95% confidence interval 0.013 to 0.093). However, quality-adjusted life-years over 26 weeks were not significantly greater (difference 0.0013, 95% confidence interval –0.0157 to 0.0182). Costs were lower in the intervention arm but, again, not significantly (–£163, 95% confidence interval –£349 to £28). Cost-effectiveness and cost–utility analyses, therefore, suggested that the intervention was dominant over usual care, but with considerable uncertainty around the point estimates. Patients valued using the Patient Health Questionnaire-9 to compare scores at baseline and follow-up, whereas practitioner views were more mixed, with some considering it too time-consuming. Conclusions We found no evidence of improved depression management or outcome at 12 weeks from using the Patient Health Questionnaire-9, but patients’ quality of life was better at 26 weeks, perhaps because feedback of Patient Health Questionnaire-9 scores increased their awareness of improvement in their depression and reduced their anxiety. Further research in primary care should evaluate patient-reported outcome measures including anxiety symptoms, administered remotely, with algorithms delivering clear recommendations for changes in treatment. Study registration This study is registered as IRAS250225 and ISRCTN17299295. Funding This award was funded by the National Institute for Health and Care Research (NIHR) Health Technology Assessment programme (NIHR award ref: 17/42/02) and is published in full in Health Technology Assessment; Vol. 28, No. 17. See the NIHR Funding and Awards website for further award information. Plain language summary Depression is common, can be disabling and costs the nation billions. The National Health Service recommends general practitioners who treat people with depression use symptom questionnaires to help assess whether those people are getting better over time. A symptom questionnaire is one type of patient-reported outcome measure. Patient-reported outcome measures appear to benefit people having therapy and mental health care, but this approach has not been tested thoroughly in general practice. Most people with depression are treated in general practice, so it is important to test patient-reported outcome measures there, too. In this study, we tested whether using a patient-reported outcome measure helps people with depression get better more quickly. The study was a ‘randomised controlled trial’ in general practices, split into two groups. In one group, people with depression completed the Patient Health Questionnaire, or ‘PHQ-9’, patient-reported outcome measure, which measures nine symptoms of depression. In the other group, people with depression were treated as usual without the Patient Health Questionnaire-9. We fed the results of the Patient Health Questionnaire-9 back to the people with depression themselves to show them how severe their depression was and asked them to discuss the results with the practitioners looking after them. We found no differences between the patient-reported outcome measure group and the control group in their level of depression; their work or social life; their satisfaction with care from their practice; or their use of medicines, therapy or specialist care for depression. However, we did find that their quality of life was improved at 6 months, and the costs of the National Health Service services they used were lower. Using the Patient Health Questionnaire-9 can improve patients’ quality of life, perhaps by making them more aware of improvement in their depression symptoms, and less anxious as a result. Future research should test using a patient-reported outcome measure that includes anxiety and processing the answers through a computer to give practitioners clearer advice on possible changes to treatment for depression. Scientific summary Some text in this chapter has been adapted from the study protocol published as: Kendrick T, Moore M, Leydon G, et al. Patient-reported outcome measures for monitoring primary care patients with depression (PROMDEP): study protocol for a randomised controlled trial. Trials 2020;21:441. https://doi.org/10.1186/s13063-020-04344-9. This article is published under license to BioMed Central Ltd. This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/4.0), which permits unrestricted use, distribution, and reproduction in any medium provided the original work is properly credited. The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article unless otherwise stated. Background Depression is common and costly. It can lead to chronic disability, poor quality of life, suicide, and high service use and costs. National Institute for Health and Care Excellence guidelines recommend different treatments for more severe and less severe depression, but general practitioners, who treat more the majority of people with depression in primary care, are often inaccurate in their global clinical assessments of depression severity, and treatment is not targeted to patients most likely to benefit. The National Institute for Health and Care Excellence recommends that practitioners consider using validated patient-reported outcome measures to inform treatment at diagnosis and follow-up of people with depression, but there is insufficient evidence that these measures improve depression management and outcomes for patients in primary care. Aim and objectives The aim of the study was to answer the research question: What is the effectiveness and cost-effectiveness of assessing primary care patients with depression or low mood soon after diagnosis and again at follow-up 10–35 days later, using the Patient Health Questionnaire-9 combined with patient feedback and practitioner guidance on treatment? The objectives were to (1) carry out a cluster-randomised controlled trial to compare the intervention with usual care; (2) provide intervention arm patients with written feedback on their Patient Health Questionnaire-9 scores, indicating evidence-based treatments relevant to the level of severity of depression to discuss with practitioners; (3) train practitioners to interpret Patient Health Questionnaire-9 scores and their implications for choice of treatment, taking into account contextual factors; (4) follow up participants for 26 weeks, with research assessments at 12 and 26 weeks; (5) determine the primary outcome of depressive symptoms on the Beck Depression Inventory, 2nd edition, at 12-week follow-up; (6) examine secondary outcomes, including depressive symptoms on the Beck Depression Inventory, 2nd edition, at 26 weeks, and social functioning and quality of life at both 12- and 26-week follow-ups; (7) measure patient satisfaction, adverse events, antidepressant treatment, secondary care contacts, service use, and costs over 26 weeks’ follow-up, and perform cost-effectiveness and cost–utility analyses; and (8) carry out a qualitative process analysis to explore participants’ reflections on the use of the Patient Health Questionnaire-9 and the potential for implementing it in practice. Methods The study design was a parallel-group, cluster-randomised superiority trial with 1 : 1 allocation to intervention and control arms. The setting was UK primary care (141 group general practices in England and Wales). Inclusion criteria were age ≥ 18 years with a new episode of depressive disorder or symptoms. Patients were recruited mainly through regular medical records searches but also opportunistically at consultations for new episodes of depression. Exclusion criteria were current treatment for depression; dementia; psychosis; substance misuse; or a significant risk of suicide. The intervention was administration of the Patient Health Questionnaire-9 questionnaire as a PROM soon after diagnosis and at follow-up 10–35 days later. Patients were given written feedback on their Patient Health Questionnaire-9 scores and potential treatments to discuss with their general practitioners. Practitioners were trained in interpreting Patient Health Questionnaire-9 scores and taking them into account in treatment decisions. The primary outcome was depressive symptoms on the Beck Depression Inventory, 2nd edition, at 12 weeks. Secondary outcomes were Beck Depression Inventory, 2nd edition, scores at 26 weeks; social functioning (on the Work and Social Adjustment Scale) and quality of life (on the EuroQol-5 Dimensions, five-level) at 12 and 26 weeks; service use including antidepressant treatment and primary and secondary care contacts over 26 weeks to calculate NHS costs; and patient satisfaction at 26 weeks (on the Medical Informant Satisfaction Scale). For our sample size calculation, we assumed a baseline mean Beck Depression Inventory, 2nd edition, score of 24.0 with a standard deviation of 10.0 (derived from a feasibility study), and mean scores at 12-week follow-up of 14.0 in the intervention arm and 17.0 in the control arm. The anticipated difference of 3.0 points (effect size of 0.3) represented the minimum clinically important difference on the Beck Depression Inventory, 2nd edition. At the 5% level of significance, to have 90% power to detect that difference we calculated we needed 235 patients analysed per arm. We aimed to recruit a mean of six patients per practice and assumed an intracluster correlation coefficient of 0.03 (from the feasibility study), which gave a cluster design effect of 1.15, meaning we needed 270 per arm. We assumed a 20% loss to follow-up at 12 weeks, so the total sample size needed was 270 × 2/0.8 and our original target sample size was a total of 676 patients recruited, from 113 practices, by three recruitment centres (the University of Southampton, the University of Liverpool and University College London). We subsequently revised the target sample size on finding a significant correlation coefficient of > 0.5 between baseline and follow-up values for the primary outcome, which meant that we needed only 222 patients analysed per arm and, therefore, a target sample size of 554 patients recruited (revised 10 June 2021). Cluster randomisation of practices to intervention and control arms was carried out remotely by a Clinical Trials Unit statistician using computerised sequence generation, with minimisation by recruiting centre, size of practice and urban or rural location. Blinding of participating practitioners and patients to allocation was impossible given the nature of the intervention and the cluster-randomised design, but self-report outcome measures were used to prevent researcher rating bias, and statistical analysis was blind to allocation. Differences between intervention and control arms in the outcomes of depressive symptoms, social functioning and quality of life measured at 12- and 26-week follow-up were analysed using linear mixed models, adjusting for baseline depression; duration of depression; history of depression; baseline anxiety; sociodemographic factors (gender, age, socioeconomic position, housing, education, marital status and dependants), and clustering including a random effect for practice. Patient satisfaction, quality of life (quality-adjusted life-years) and costs were compared between the arms over the 26 weeks’ study follow-up period. Differences between the arms in the process of care for depression were also analysed, including patients’ self-reported use of antidepressants at the 12- and 26-week follow-up points, and medication and contacts with mental health services (community mental health nurses, counsellors, psychologists, psychiatrists, other therapists and social workers) recorded in practice medical records over the 26 weeks’ follow-up. A health economic evaluation was undertaken from an NHS and Personal Social Services perspective. The outcomes were expressed as incremental cost per point improvement in the Beck Depression Inventory, 2nd edition, clinical outcome (cost-effectiveness analysis), and incremental cost per quality-adjusted life-year gained (cost–utility analysis). The primary analysis at 26 weeks used a generalised linear mixed model to estimate the differences in costs and quality-adjusted life-years (using the EuroQol-5 Dimensions, five-level to calculate patient utilities), adjusted for baseline quality of life; baseline anxiety; sociodemographic factors; and practice as a random effect. Incremental cost-effectiveness ratios and a cost-effectiveness acceptability curve were generated using non-parametric bootstrapping. Qualitative interviews with participating practitioners and patients in both arms were conducted to reflect on their involvement in the trial and analysed using reflexive thematic analysis. Intervention arm participants were asked about barriers, facilitators, benefits and problems related to using the Patient Health Questionnaire-9, including questions derived from the normalisation process theory framework. Results Practices and patients As the number of patients recruited per practice was smaller than anticipated, we recruited significantly more than our target of 113 practices, eventually reaching a total of 189, but 48 practices subsequently withdrew (24 in each arm), so the final number of active practices was 141: 72 intervention and 69 control (28 above our original target). Practice characteristics were well balanced by arm. Of 11,468 patients approached in consultations or through mailed invitations, 1058 (9.2%) returned reply slips about the study: 574 (10.6% of those approached) in the intervention arm and 484 (8.0% of those approached) in the control arm. After the exclusion of patients declining to participate, ineligible at screening or uncontactable, 529 patients were assessed at baseline: 302 (5.5% of those approached) in the intervention arm and 227 (3.8% of those approached) in the control arm. The ratio of intervention to control arm patients recruited was, therefore, 1.3 to 1, which may have reflected lower motivation to take part among control arm practices. Of 529 patients recruited, 453 (85.6%) were followed up at 12 weeks: 254 intervention arm (84.1%) and 199 control arm (87.7%) patients. At the 26-week point, 414 patients (78.3%) were followed up: 230 intervention arm (76.2%) and 184 control arm (81.1%). Medical records data were collected for 259 intervention arm patients (85.8%) and 201 control arm patients (88.5%). The mean BDI-II score for depressive symptoms at baseline was higher in the intervention arm, at 24.1 (standard deviation 8.89) than in the control arm, at 22.4 (standard deviation 9.52). Baseline anxiety and quality-of-life scores were also worse in the intervention arm. Control arm patients were more likely to have had two or more previous depressive episodes. Demographic characteristics were relatively well balanced. Clinical outcomes At the 12-week follow-up, the mean Beck Depression Inventory, 2nd edition, score was 18.5 (standard deviation 10.2) in the intervention arm and 16.9 (standard deviation 10.3) in the control arm. The adjusted mean score was slightly lower in the intervention arm, but this was not statistically significant (mean adjusted difference –0.46, 95% confidence interval –2.16 to 1.26; p = 0.60). At 26 weeks, the mean Beck Depression Inventory, 2nd edition, scores were 15.1 (standard deviation 10.8) in the intervention arm and 14.7 (standard deviation 10.6) in the control arm (mean adjusted difference –1.63, 95% confidence interval –3.48 to 0.21; p = 0.08). Social functioning on the Work and Social Adjustment Scale and Medical Informant Satisfaction Scale satisfaction with care scores favoured the intervention, but the differences found were not statistically significant. A post hoc analysis at 26 weeks showed similar proportions improving by ≥ 50% on the Beck Depression Inventory, 2nd edition, in the intervention and control arms (45.1% vs. 37.3%), but the proportion remitting to a score of
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- 2024
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15. Abstract 126: HUB Organoids: bringing the 'patient in the lab' for preclinical and clinical development
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Jasper Sluimer, Lars-Eric Fielmich, Robert G.J. Vries, Jasper Mullenders, Jane Sun, Annemarie Buijs, and Sylvia F. Boj
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Oncology ,Cancer Research ,medicine.medical_specialty ,business.industry ,Cancer ,Disease ,Patient specific ,medicine.disease ,Biobank ,Molecular level ,Internal medicine ,Organoid ,Medicine ,Molecular Profile ,Personalized medicine ,business - Abstract
Preclinical disease models are essential for the development of novel therapies and help better understand the molecular processes. In recent years, the groundbreaking technology to generate patient specific in vitro cultures based on adult stem-cell (ASC) organoids, has gained widespread interest for the development of new therapies and as a predictive diagnostic tool. ASC-derived organoids can be generated at high efficiency from both healthy and diseased tissue (resections as well as biopsies) including most carcinomas. These patient-derived cultures can be expanded for prolonged periods of time while maintaining many properties of the tissue they were derived from. At Hubrecht Organoid Technology (HUB), we have created comprehensive ‘organoid living biobanks' from tumors of different origin like colon, lung, breast, pancreas and ovary. These biobanks have been characterized at the genetic and transcriptional level. These analyses have shown that our ‘organoid living biobanks' capture disease heterogeneity at the genetic and molecular level. Because HUB Organoids can be expanded long term, drug screening is feasible in these models. HUB has developed procedures for (high-throughput) compound screening in organoids including small molecules and biologics. In this setting, HUB Organoids have been used to identify novel targets, identify lead compounds and stratify potential responders based on their genetic or molecular profile. In addition, HUB Organoids can serve as a patient avatar in clinical development or diagnostics. In this setting, HUB Organoids could add a functional test for therapy response in a patient centered setting providing a unique tool for personalized medicine. Citation Format: Jasper Mullenders, Annemarie Buijs, Lars-Eric Fielmich, Jasper Sluimer, Jane Sun, Robert G. Vries, Sylvia F. Boj. HUB Organoids: bringing the 'patient in the lab' for preclinical and clinical development [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2021; 2021 Apr 10-15 and May 17-21. Philadelphia (PA): AACR; Cancer Res 2021;81(13_Suppl):Abstract nr 126.
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- 2021
16. TWIST1 homodimers and heterodimers orchestrate lineage-specific differentiation
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Mark E. Graham, Jane Sun, Pierre Osteil, Ashley J. Waardenberg, Madeleine Demuth, Xiaochen Fan, David A.F. Loebel, Nicolas Fossat, and Patrick P.L. Tam
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0303 health sciences ,Mesoderm ,Lineage (genetic) ,animal structures ,Mesenchyme ,Neural crest ,Biology ,Embryonic stem cell ,Cell biology ,03 medical and health sciences ,0302 clinical medicine ,medicine.anatomical_structure ,030220 oncology & carcinogenesis ,TCF3 ,medicine ,Stem cell ,Endoderm ,030304 developmental biology - Abstract
The extensive array of bHLH transcription factors and their combinations as dimers underpin the diversity of molecular function required for cell type specification during embryogenesis. The bHLH factor TWIST1 plays pleiotropic roles during development. However, which combinations of TWIST1 dimers are involved and what impact each dimer imposes on the gene regulation network controlled by TWIST1 remain elusive. In this work, proteomic profiling of human-TWIST1 expressing cell lines and transcriptome analysis of mouse cranial mesenchyme have revealed that TWIST1 homodimer and heterodimers with TCF3, TCF4 and TCF12 E-proteins are the predominant dimer combinations. Dimers formation or their balance are altered by disease-causing mutations in TWIST1 helix domains, which may account for the defective differentiation of the craniofacial mesenchyme observed in patients. Functional analyses of the loss and gain of TWIST1-E-protein dimer activity have revealed previously unappreciated roles in guiding lineage differentiation of embryonic stem cells: TWIST1-E-protein heterodimers activate the differentiation of mesoderm and neural crest cells which is accompanied by epithelial-to-mesenchymal transition, while TWIST1 homodimers maintain the stem cells in a progenitor state and block entry to the endoderm lineage.
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- 2019
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17. Gene Editing of Mouse Embryonic and Epiblast Stem Cells
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Tennille, Sibbritt, Pierre, Osteil, Xiaochen, Fan, Jane, Sun, Nazmus, Salehin, Hilary, Knowles, Joanne, Shen, and Patrick P L, Tam
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Gene Editing ,Gene Knockout Techniques ,Mice ,Animals ,Clustered Regularly Interspaced Short Palindromic Repeats ,Mouse Embryonic Stem Cells ,CRISPR-Cas Systems ,Frameshift Mutation ,Cells, Cultured ,Germ Layers ,Plasmids ,RNA, Guide, Kinetoplastida - Abstract
Efficient and reliable methods for gene editing are critical for the generation of loss-of-gene function stem cells and genetically modified mice. Here, we outline the application of CRISPR-Cas9 technology for gene editing in mouse embryonic stem cells (mESCs) to generate knockout ESC chimeras for the fast-tracked analysis of gene function. Furthermore, we describe the application of gene editing directly to mouse epiblast stem cells (mEpiSCs) for modelling germ layer differentiation in vitro.
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- 2019
18. Integrated therapist and online CBT for depression in primary care (INTERACT): study protocol for a multi-centre randomised controlled trial
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Debbie Tallon, Laura Thomas, Sally Brabyn, Brian Chi Fung Ching, Jane Sungmin Hahn, Berry Jude, Mekeda X Logan, Alex Burrage, Fiona Fox, Simon Gilbody, Paul Lanham, Glyn Lewis, Jinshuo Li, Stephanie J. MacNeill, Irwin Nazareth, Steve Parrott, Tim J. Peters, Roz Shafran, Katrina Turner, Chris Williams, David Kessler, and Nicola Wiles
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Depression ,Randomised controlled trial ,Cognitive behavioural therapy ,Internet-based treatment ,Blended treatment ,Primary care ,Medicine (General) ,R5-920 - Abstract
Abstract Background Cognitive behavioural therapy (CBT) is an effective treatment for depression. Self-directed online CBT interventions have made CBT more accessible at a lower cost. However, adherence is often poor and, in the absence of therapist support, effects are modest and short-term. Delivering CBT online using instant messaging is clinically and cost-effective; however, most existing platforms are limited to instant messaging sessions, without the support of between-session “homework” activities. The INTERACT intervention integrates online CBT materials and ‘high-intensity’ therapist-led CBT, delivered remotely in real-time. The INTERACT trial will evaluate this novel integration in terms of clinical and cost-effectiveness, and acceptability to therapists and clients. Methods Pragmatic, two parallel-group multi-centre individually randomised controlled trial, with 434 patients recruited from primary care practices in Bristol, London and York. Participants with depression will be identified via General Practitioner record searches and direct referrals. Inclusion criteria: aged ≥ 18 years; score ≥ 14 on Beck Depression Inventory (BDI-II); meeting International Classification of Diseases (ICD-10) criteria for depression. Exclusion criteria: alcohol or substance dependency in the past year; bipolar disorder; schizophrenia; psychosis; dementia; currently under psychiatric care for depression (including those referred but not yet seen); cannot complete questionnaires unaided or requires an interpreter; currently receiving CBT/other psychotherapy; received high-intensity CBT in the past four years; participating in another intervention trial; unwilling/unable to receive CBT via computer/laptop/smartphone. Eligible participants will be randomised to integrated CBT or usual care. Integrated CBT utilises the standard Beckian intervention for depression and comprises nine live therapist-led sessions, with (up to) a further three if clinically appropriate. The first session is 60–90 min via videocall, with subsequent 50-min sessions delivered online, using instant messaging. Participants allocated integrated CBT can access integrated online CBT resources (worksheets/information sheets/videos) within and between sessions. Outcome assessments at 3-, 6-, 9- and 12-month post-randomisation. The primary outcome is the Beck Depression Inventory (BDI-II) score at 6 months (as a continuous variable). A nested qualitative study and health economic evaluation will be conducted. Discussion If clinically and cost-effective, this model of integrated CBT could be introduced into existing psychological services, increasing access to, and equity of, CBT provision. Trial registration ISRCTN, ISRCTN13112900. Registered on 11/11/2020. Currently recruiting participants. Trial registration data are presented in Table 1.
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- 2023
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19. How frequent is routine use of probiotics in UK neonatal units?
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Justinas Teiserskas, Rachel Hutchinson, Lisa Szatkowski, Claire Caldwell, Katie Taylor, Emilie Seager, Catherine Longley, Rebecca Smith, Brandy Cox, Cheryl Battersby, Helen Lloyd, Aneurin Young, Deborah Davidson, Jennifer Peterson, Emma Williams, Kirti Gupta, Ahmed Mohamed, Paul Fleming, Tal Oryan, Mia Kahvo, Arameh Aghababaie, Janet Berrington, Michelle Fernandes, Neaha Patel, Jessica Farnan, Allan Jenkinson, Bushra Abdul-Malik, Lucinda Winckworth, Kate Costeloe, Christopher Freeman, Katie Evans, Jasmine Taylor, Mary-Rose Ballard, Rhiannon Jones, Rajkumar Dhandayuthapani, Caroline Fraser, James Stevens, Nuala Calder, Amy Grant, Moataz Badawy, Afza Sadiq, Manohar Joishy, Nathan Collicott, Naseem Sharif, Spandana Rupa Madabhushi, G Natasha, Joe McConville, Rhianna Netherton, Lizaveta Collins, Naomi Lin, Kouros Driscoll, Jonathan Talbot, Rosie Roots, Alison Hopper, Camilla James, Shreesh Bhat, Lauren Ferretti, Niha Peshimam, Benjamin Holter, Sion Glaze, Anna Waghorn, Shweta Dixit, Chibuko Ukeje, Shana Irvine, Fergus Harnden, Christine Lim, Neelakshi Ghosh, Eileen Foster, Swati Jha, Joanna O’Sullivan, Evangelia Myttaraki, Shreya Agrawal, Steve Abbey, Abdulhakim Abdurrazaq, Saud Ahmed, Faith Akano, Muhammad Rehan Akhtar, Oghenetekevwe Patrick Akpofure, Myriam Segovia Almiron, Namita Anand, Jessica Archibald, Harriet Aughey, Lynnlette Aung, Thandi Aung, Pramila Bade, Naomi Bell, Andrada Maria Bianu, Catherine Black, Gennie Booth, Karla Buerano, Chinnu Chandran, Shavin Chellen, Ruth Cousins, Leanne Dearman, Alshaimaa Eldeeb, Teim Eyo, Yasin Fatine, Poppy Flanagan, Abhrajit Giri, Saqib Hasan, Craig Haverstock, Jayne Hillier, Kate Hooper, Zoe Howard, Mais Ismail, Matilda Iverson, Sam Jay, Katie Jenkins, Carla Kantyka, Caroline Kargbo, Almutassem Kazkaz, Shelley Knights, Nikoletta Kottarakou, Carianne Lewis, Carys Mangan, Diane McCarter, Aodhan McGillian, Tasneem Modan, Maria Orford, Salil Pradhan, Patrycja Prusak, Ayesha Rahim, Daniel Ratnaraj, Naveed Shahzad, Adwa Shalabi, Claire Strauss, Jane Sundarsingh, Sumit Thankur, Toby Thenat, Alice Unsworth, Carl Van Heyningen, and Elena Raluka Vlad
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Pediatrics ,RJ1-570 - Abstract
Objective There is a lack of UK guidance regarding routine use of probiotics in preterm infants to prevent necrotising enterocolitis, late-onset sepsis and death. As practices can vary, we aimed to determine the current usage of probiotics within neonatal units in the UK.Design and setting Using NeoTRIPS, a trainee-led neonatal research network, an online survey was disseminated to neonatal units of all service levels within England, Scotland, Northern Ireland and Wales in 2022. Trainees were requested to complete one survey per unit regarding routine probiotic administration.Results 161 of 188 (86%) neonatal units responded to the survey. 70 of 161 (44%) respondents routinely give probiotics to preterm infants. 45 of 70 (64%) use the probiotic product Lactobacillus acidophilus NCFM/Bifidobacterium bifidum Bb-06/B. infantis Bi-26 (Labinic™). 57 of 70 (81%) start probiotics in infants ≤32 weeks’ gestation. 33 of 70 (47%) had microbiology departments that were aware of the use of probiotics and 64 of 70 (91%) had a guideline available. Commencing enteral feeds was a prerequisite to starting probiotics in 62 of 70 (89%) units. The majority would stop probiotics if enteral feeds were withheld (59 of 70; 84%) or if the infant was being treated for necrotising enterocolitis (69 of 70; 99%). 24 of 91 (26%) units that did not use probiotics at the time of the survey were planning to introduce them within the next 12 months.Conclusions More than 40% of all UK neonatal units that responded are now routinely administering probiotics, with variability in the product used. With increased probiotic usage in recent years, there is a need to establish whether this translates to improved clinical outcomes.
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- 2023
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20. ‘The phoenix that always rises from the ashes’: an exploratory qualitative study of the experiences of an initiative informed by principles of psychological first aid following the Beirut blast
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Brian Chi Fung Ching, Alexandra Badaoui, Nada Abou Seif, Rea Al Hallal, Gabriel Luiz Bundies, Amy Campbell, Ayla Rafie, Angela Song-Chase, Jane Sungmin Hahn, and Jo Billings
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Beirut blast ,remote support ,psychological first aid ,qualitative ,trauma ,Explosión de Beirut ,Psychiatry ,RC435-571 - Abstract
ABSTRACTBackground: On 4 August 2020, an explosion occurred in Beirut, Lebanon. Hundreds of people were killed, thousands injured and displaced. An initiative was rapidly initiated to provide remote support informed by psychological first aid for the mental health of Lebanese young adults affected by the blast. However, little is known about recipients’ experiences of such initiatives.Objective: This study aimed to qualitatively explore the experiences of supporters and recipients in the community-led initiative following the blast.Method: We recruited a diverse sample of four supporters and four Lebanese recipients who took part in the Beirut initiative. Semi-structured interviews were conducted with participants. Reflexive thematic analysis was used to analyse the qualitative data.Results: We developed five themes from the qualitative interviews, which highlighted ideas around accessibility, alienation, the relationship, elements of the safe space created by the initiative, and unmet needs and areas for improvement. Recipients described the detrimental impact of the blast on their mental health within the Lebanese context and beyond. Recipients and supporters elucidated complex experiences of the support and its impact.Conclusions: Our findings suggest remote support has the potential to be acceptable for young adults in Lebanon. Further research into support informed by psychological first aid after similar crisis events is warranted.
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- 2023
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21. Industrial Applications of Electron Microscopy: A Shared Laboratory Perspective
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Peng Zhang, Jane Sun, Shaojie Wang, Jingyi Zhang, M. E. Salmon, and Mark Izquierdo
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Materials science ,law ,Perspective (graphical) ,Nanotechnology ,Electron microscope ,Instrumentation ,law.invention - Published
- 2019
22. Transcriptional evaluation of the developmental accuracy, reproducibility and robustness of kidney organoids derived from human pluripotent stem cells
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Ernst J. Wolvetang, Minoru Takasato, Pei Xuan Er, Alicia Oshlack, Belinda Phipson, Jane Sun, David Yen, Kynan T. Lawlor, Melissa H. Little, and Lorna J Hale
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0303 health sciences ,Robustness (evolution) ,Nephron ,Biology ,Epithelium ,3. Good health ,Cell biology ,03 medical and health sciences ,0302 clinical medicine ,Directed differentiation ,medicine.anatomical_structure ,medicine ,Organoid ,Induced pluripotent stem cell ,Gene ,Functional genomics ,030217 neurology & neurosurgery ,030304 developmental biology - Abstract
We have previously reported a protocol for the directed differentiation of human induced pluripotent stem cells to kidney organoids comprised of nephrons, proximal and distal epithelium, vasculature and surrounding interstitial elements. The utility of this protocol for applications such as disease modelling will rely implicitly on the developmental accuracy of the model, technical robustness of the protocol and transferability between iPSC lines. Here we report extensive transcriptional analyses of the sources of variation across the timecourse of differentiation from pluripotency to complete kidney organoid, focussing on repeated differentiations to day 18 organoid. Individual organoids generated within the same differentiation experiment show Spearman’s correlation coefficients of >0.99. The greatest source of variation was seen between experimental batch, with the enrichment for genes that also varied temporally between day 10 and day 25 organoids implicating nephron maturation as contributing to transcriptional variance between individual differentiation experiments. A morphological analysis revealed a transition from renal vesicle to capillary loop stage nephrons across the same time period. Distinct iPSC clones were also shown to display congruent transcriptional programs with inter-experimental and inter-clonal variation most strongly associated with nephron patterning. Even epithelial cells isolated from organoids showed transcriptional alignment with total organoids of the same day of differentiation. This data provides a framework for managing experimental variation, thereby increasing the utility of this approach for personalised medicine and functional genomics.
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- 2017
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23. Generation and Characterization of Leukemia Inhibitory Factor-Dependent Equine Induced Pluripotent Stem Cells from Adult Dermal Fibroblasts
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Ernst J. Wolvetang, Deanne J. Whitworth, Dmitry A. Ovchinnikov, Patrick R.J. Fortuna, and Jane Sun
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Homeobox protein NANOG ,X Chromosome ,Rex1 ,Induced Pluripotent Stem Cells ,Basic fibroblast growth factor ,Gene Expression ,Embryoid body ,Biology ,Fibroblast growth factor ,Leukemia Inhibitory Factor ,Histones ,chemistry.chemical_compound ,Original Research Reports ,Animals ,Horses ,Induced pluripotent stem cell ,Cell Proliferation ,Skin ,Feeder Cells ,Cell Biology ,Hematology ,Fibroblasts ,Molecular biology ,Coculture Techniques ,Fibroblast Growth Factors ,chemistry ,embryonic structures ,Female ,Reprogramming ,Leukemia inhibitory factor ,Biomarkers ,Transcription Factors ,Developmental Biology - Abstract
In this study we have reprogrammed dermal fibroblasts from an adult female horse into equine induced pluripotent stem cells (equiPSCs). These equiPSCs are dependent only on leukemia inhibitory factor (LIF), placing them in striking contrast to previously derived equiPSCs that have been shown to be co-dependent on both LIF and basic fibroblast growth factor (bFGF). These equiPSCs have a normal karyotype and have been maintained beyond 60 passages. They possess alkaline phosphatase activity and express eqNANOG, eqOCT4, and eqTERT mRNA. Immunocytochemistry confirmed that they produce NANOG, REX1, SSEA4, TRA1-60, and TRA1-81. While our equiPSCs are LIF dependent, bFGF co-stimulates their proliferation via the PI3K/AKT pathway. EquiPSCs lack expression of eqXIST and immunostaining for H3K27me3, suggesting that during reprogramming the inactive X chromosome has likely been reactivated to generate cells that have two active X chromosomes. EquiPSCs form embryoid bodies and in vitro teratomas that contain derivatives of all three germ layers. These LIF-dependent equiPSCs likely reflect a more naive state of pluripotency than equiPSCs that are co-dependent on both LIF and bFGF and so provide a novel resource for understanding pluripotency in the horse.
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- 2014
24. Retardation of Grain Growth and Grain Boundary Pinning in Athermal Block Copolymer Blend Systems
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Apostolos Avgeropoulos, Michael R. Bockstaller, Hyung Ju Ryu, and Jane Sun
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Materials science ,Polymers and Plastics ,Organic Chemistry ,Grain size ,Inorganic Chemistry ,Grain growth ,visual_art ,Particle-size distribution ,Materials Chemistry ,visual_art.visual_art_medium ,Grain boundary diffusion coefficient ,Effective diffusion coefficient ,Grain boundary ,Ceramic ,Composite material ,Grain boundary strengthening - Abstract
The effect of filler addition on the grain coarsening characteristics of block copolymer materials is analyzed for the particular case of a lamellar poly(styrene-b-isoprene)-type block copolymer and polystyrene as well as polystyrene-grafted nanoparticle fillers. Filler addition is shown to reduce the rate of grain growth and to induce grain size distributions that deviate from the log-normal type that is characteristic for pristine block copolymer systems. The retardation of grain growth is shown to be associated with the segregation of filler additives into high energy grain boundary defects—a process that bears similarities to the segregation of impurity atoms within grain boundary structures in ceramics or metals. The analysis of grain boundary energy, grain size distribution, and grain coarsening kinetics suggests two major mechanisms for the interference of filler additives with grain coarsening: First, the segregation of fillers into boundary regions lowers the relative grain boundary energy and he...
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- 2014
25. Generation of Footprint-Free Induced Pluripotent Stem Cells from Human Fibroblasts Using Episomal Plasmid Vectors
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Dmitry A, Ovchinnikov, Jane, Sun, and Ernst J, Wolvetang
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Induced Pluripotent Stem Cells ,Cell Culture Techniques ,Gene Expression ,Humans ,Transgenes ,Fibroblasts ,Cellular Reprogramming ,Transfection ,Polymerase Chain Reaction ,Plasmids - Abstract
Human induced pluripotent stem cells (hiPSCs) have provided novel insights into the etiology of disease and are set to transform regenerative medicine and drug screening over the next decade. The generation of human iPSCs free of a genetic footprint of the reprogramming process is crucial for the realization of these potential uses. Here we describe in detail the generation of human iPSC from control and disease-carrying individuals' fibroblasts using episomal plasmids.
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- 2015
26. The Rho Guanosine 5′-Triphosphatase, Cell Division Cycle 42, Is Required for Insulin-Induced Actin Remodeling and Glucagon-Like Peptide-1 Secretion in the Intestinal Endocrine L Cell
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Patricia L. Brubaker, Jane Sun, Molie Xu, Tianru Jin, and Gareth E. Lim
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MAPK/ERK pathway ,medicine.medical_specialty ,Cell division ,Insulin ,medicine.medical_treatment ,Actin remodeling ,Enteroendocrine cell ,RAC1 ,macromolecular substances ,CDC42 ,Biology ,Endocrinology ,Internal medicine ,medicine ,Secretion - Abstract
Rho GTPases, such as cell division cycle 42 (Cdc42) and ras-related C3 botulinum toxin substrate 1 (Rac1), have been identified as regulators of F-actin dynamics and hormone release from endocrine cells; however, their role in secretion of the incretin hormone, glucagon-like peptide-1 (GLP-1), from the enteroendocrine L cell is unknown. Insulin induced a 1.4-fold increase in L cell GLP-1 release; however, secretion was potentiated to 2.1-fold in the presence of the F-actin depolymerizing agent, latrunculin B, suggesting that F-actin functions as a permissive barrier. In murine GLUTag L cells, insulin stimulated F-actin depolymerization and Cdc42 activation simultaneously, and these events occurred prior to detectable increases in insulin-induced GLP-1 release. After insulin treatment, Cdc42-dependent p21-activated kinase-1 (PAK1) activation was also detected, and transfection of small-interfering RNA against Cdc42 or of dominant-negative Cdc42(T17N) impaired insulin-stimulated PAK1 activation, actin remodeling, and GLP-1 secretion. Overexpression of kinase-dead PAK1(K299R) or PAK1 small interfering RNA similarly attenuated insulin-induced GLP-1 secretion. Knockdown or inhibition of Cdc42 and PAK1 activities also prevented activation of MAPK/ERK (MEK)-1/2-ERK1/2 by insulin, which was previously identified as a critical pathway for insulin-regulated GLP-1 release. Taken together, these data identify a novel signaling pathway in the endocrine L cell, whereby Cdc42 regulates actin remodeling, activation of the cannonical 1/2-ERK1/2 pathway and PAK1, and GLP-1 secretion in response to insulin.
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- 2009
27. Both Wnt and mTOR signaling pathways are involved in insulin-stimulated proto-oncogene expression in intestinal cells
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Tianru Jin and Jane Sun
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Beta-catenin ,medicine.medical_treatment ,Proto-Oncogene Mas ,Cell Line ,Proto-Oncogene Proteins c-myc ,Phosphatidylinositol 3-Kinases ,Cell Line, Tumor ,Hyperinsulinism ,medicine ,Animals ,Humans ,Insulin ,Intestinal Mucosa ,Protein kinase B ,beta Catenin ,PI3K/AKT/mTOR pathway ,Cell Proliferation ,biology ,Cell growth ,TOR Serine-Threonine Kinases ,Wnt signaling pathway ,Cell Biology ,Rats ,Cell biology ,Intestines ,Wnt Proteins ,Insulin receptor ,Crosstalk (biology) ,biology.protein ,Colorectal Neoplasms ,Protein Kinases ,Proto-Oncogene Proteins c-akt ,Signal Transduction - Abstract
Subjects with Type II diabetes mellitus are more vulnerable in developing colorectal tumors, suggesting that hyperinsulinemia may stimulate proto-oncogene expression, and the existence of crosstalk between insulin signaling and pathways that are involved in colorectal tumor formation. We show here that insulin stimulates cell proliferation and c-Myc expression in colon cancer cell lines HT29 and Caco-2, intestinal non-cancer cell line IEC-6, and primary fetal rat intestinal cell (FRIC) cultures. The effect of insulin was blocked by phosphoinositide-3 Kinase (PI3K) inhibition, but only partially attenuated by inhibition of Protein kinase B (PKB), indicating the existence of both PKB-dependent and -independent mechanisms. The PKB-dependent mechanism of insulin-stimulated c-Myc expression in HT29 cells was shown to involve the activation of mTOR in c-Myc translation. In the investigation of the PKB-independent mechanism, we found that insulin-induced nuclear translocation of β-catenin (β-cat), an effector of Wnt signaling. Furthermore, insulin stimulated the expression of TopFlash, a Wnt-responsive reporter gene. Finally, chromatin immunoprecipitation (ChIP) detected significant increases in the binding of β-cat to two TCF binding sites of the human c-Myc promoter following insulin treatment. Our observations support the existence of crosstalk between insulin and Wnt signaling pathways, and suggest that the crosstalk involves a PKB-independent mechanism.
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- 2008
28. Inhibition of Airway Na + Transport by Respiratory Syncytial Virus
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David I. Cook, Karl Kunzelmann, Jane Sun, Nicholas J. C. King, and Jayesh Meanger
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Epithelial sodium channel ,Respiratory Mucosa ,viruses ,Immunology ,Biology ,Microbiology ,Glycosphingolipids ,Virus ,Cell Line ,Mice ,Virology ,medicine ,Animals ,Respiratory system ,Epithelial Sodium Channels ,Protein Kinase C ,Ion Transport ,Cell Membrane ,Sodium ,Apical membrane ,medicine.disease ,Recombinant Proteins ,Respiratory Syncytial Viruses ,Toll-Like Receptor 4 ,Trachea ,medicine.anatomical_structure ,Bronchiolitis ,Insect Science ,Pathogenesis and Immunity ,Respiratory epithelium ,Viral Fusion Proteins ,Respiratory tract - Abstract
In previous studies, we have shown that two major respiratory pathogens, influenza virus and parainfluenza virus, produce acute alterations in ion transport upon contacting the apical membrane of the respiratory epithelium. In the present study, we examine the effects on ion transport by the mouse tracheal epithelium of a third major respiratory pathogen, respiratory syncytial virus (RSV). RSV infections are associated with fluid accumulation in the respiratory tract and cause illnesses that range in severity from rhinitis, sinusitis, otitis media, and bronchitis to bronchiolitis and pneumonia. We find that within minutes of RSV contacting the apical membrane; it inhibits amiloride-sensitive Na + transport by the epithelium. This effect is mediated by protein kinase C and is reproduced by recombinant viral F (fusion) protein. Since this inhibition is not accompanied by any alteration in the epithelial responses to carbachol or to forskolin plus 3-isobutyl-1-methylxanthine (IBMX), it is not due to a nonspecific toxic action of the virus. The inhibition also appears to require Toll-like receptor 4 and the presence of asialogangliosides in the apical membrane. Since the concentration range over which this inhibition is observed (10 2 to 10 5 PFU/ml) is comparable to the viral concentrations observed in clinical and experimental RSV infections, it seems likely that direct inhibition by the virus of epithelial Na + transport may contribute to the fluid accumulation that is observed in RSV infections.
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- 2007
29. HIV gp41 C-terminal Heptad Repeat Contains Multifunctional Domains
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Weiguo Jing, James Farmar, Hong Lu, Shibo Jiang, Jinkui Niu, Byron Cheung, Jane Sun, Xuxia Yan, Shuwen Liu, and Shuguang Wu
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chemistry.chemical_classification ,Peptide ,Cell Biology ,Biology ,Gp41 ,Biochemistry ,N-terminus ,Heptad repeat ,Protein structure ,chemistry ,Pepscan ,Mechanism of action ,medicine ,medicine.symptom ,Molecular Biology ,Peptide sequence - Abstract
T20 (Fuzeon), a novel anti-human immunodeficiency virus (HIV) drug, is a peptide derived from HIV-1 gp41 C-terminal heptad repeat (CHR). Its mechanism of action has not yet been defined. We applied Pepscan strategy to determine the relationship between functional domains and mechanisms of action of five 36-mer overlapping peptides with a shift of five amino acids (aa): CHR-1 (aa 623–658), C36 (aa 628–663), CHR-3 (aa 633–668), T20 (aa 638–673), and CHR-5 (aa 643–678). C36 is a peptide with addition of two aa to the N terminus of C34. Peptides CHR-1 and C36 contain N-terminal heptad repeat (NHR)- and pocket-binding domains. They inhibited HIV-1 fusion by interacting with gp41 NHR, forming stable six-helix bundles and blocking gp41 core formation. Peptide T20 containing partial NHR- and lipid-binding domains, but lacking pocket-binding domain, blocked viral fusion by binding its N- and C-terminal sequences with gp41 NHR and cell membrane, respectively. Peptide CHR-3, which is located in the middle between C36 and T20, overlaps >86% of the sequences of these two peptides, and lacks pocket- and lipid-binding domains, exhibited marginal anti-HIV-1 activity. These results suggest that T20 and C36 contain different functional domains, through which they inhibit HIV-1 entry with distinct mechanisms of action. The multiple functional domains in gp41 CHR and their binding partners may serve as targets for rational design of new anti-HIV-1 drugs and vaccines.
- Published
- 2007
30. A new model to study neurodegeneration in ataxia oculomotor apraxia type 2
- Author
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Giuseppe De Michele, Jane Sun, Martin F. Lavin, Brent L. Fogel, Chiara Criscuolo, Sam P Nayler, Olivier J. Becherel, Giovanni Coppola, Ernst J. Wolvetang, Fuying Gao, Abrey J. Yeo, Becherel, Olivier J, Sun, Jane, Yeo, Abrey J, Nayler, Sam, Fogel, Brent L, Gao, Fuying, Coppola, Giovanni, Criscuolo, Chiara, DE MICHELE, Giuseppe, Wolvetang, Ernst, and Lavin, Martin F.
- Subjects
Ataxia ,Induced Pluripotent Stem Cells ,Apoptosis ,Biology ,Mice ,Neural Stem Cells ,Genetics ,medicine ,Animals ,Humans ,Spinocerebellar Ataxias ,DNA Breaks, Double-Stranded ,Oculomotor apraxia ,Induced pluripotent stem cell ,Molecular Biology ,Genetics (clinical) ,Neurons ,Neurodegeneration ,DNA Helicases ,Autosomal recessive cerebellar ataxia ,General Medicine ,Articles ,Fibroblasts ,medicine.disease ,Cellular Reprogramming ,Multifunctional Enzymes ,Neural stem cell ,Disease Models, Animal ,Oxidative Stress ,Mutation ,Spinocerebellar ataxia ,Female ,medicine.symptom ,Neuroscience ,Reprogramming ,RNA Helicases - Abstract
Ataxia oculomotor apraxia type 2 (AOA2) is a rare autosomal recessive cerebellar ataxia. Recent evidence suggests that the protein defective in this syndrome, senataxin (SETX), functions in RNA processing to protect the integrity of the genome. To date, only patient-derived lymphoblastoid cells, fibroblasts and SETX knockdown cells were available to investigate AOA2. Recent disruption of the Setx gene in mice did not lead to neurobehavioral defects or neurodegeneration, making it difficult to study the etiology of AOA2. To develop a more relevant neuronal model to study neurodegeneration in AOA2, we derived neural progenitors from a patient with AOA2 and a control by induced pluripotent stem cell (iPSC) reprogramming of fibroblasts. AOA2 iPSC and neural progenitors exhibit increased levels of oxidative damage, DNA double-strand breaks, increased DNA damage-induced cell death and R-loop accumulation. Genome-wide expression and weighted gene co-expression network analysis in these neural progenitors identified both previously reported and novel affected genes and cellular pathways associated with senataxin dysfunction and the pathophysiology of AOA2, providing further insight into the role of senataxin in regulating gene expression on a genome-wide scale. These data show that iPSCs can be generated from patients with the autosomal recessive ataxia, AOA2, differentiated into neurons, and that both cell types recapitulate the AOA2 cellular phenotype. This represents a novel and appropriate model system to investigate neurodegeneration in this syndrome.
- Published
- 2015
31. Generation of Footprint-Free Induced Pluripotent Stem Cells from Human Fibroblasts Using Episomal Plasmid Vectors
- Author
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Ernst J. Wolvetang, Jane Sun, and Dmitry A. Ovchinnikov
- Subjects
Plasmid ,Cell culture ,Transgene ,Transfection ,Human Induced Pluripotent Stem Cells ,Biology ,Induced pluripotent stem cell ,Regenerative medicine ,Molecular biology ,Reprogramming ,Cell biology - Abstract
Human induced pluripotent stem cells (hiPSCs) have provided novel insights into the etiology of disease and are set to transform regenerative medicine and drug screening over the next decade. The generation of human iPSCs free of a genetic footprint of the reprogramming process is crucial for the realization of these potential uses. Here we describe in detail the generation of human iPSC from control and disease-carrying individuals' fibroblasts using episomal plasmids.
- Published
- 2015
32. Role of the Exchange Protein Directly Activated by Cyclic Adenosine 5′-Monophosphate (Epac) Pathway in Regulating Proglucagon Gene Expression in Intestinal Endocrine L Cells
- Author
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Zhihong Li, Tianru Jin, Shamim Lotfi, Patrick P.L. Lam, Yang Zuo, Jane Sun, Mehdi Rahimi, Youhou Kang, Herbert Y. Gaisano, Diana Islam, and Peixiang Wang
- Subjects
medicine.medical_specialty ,IBMX ,Transcription, Genetic ,MAP Kinase Signaling System ,Phosphodiesterase Inhibitors ,Enteroendocrine Cells ,Biology ,Proglucagon ,medicine.disease_cause ,Cell Line ,Mice ,chemistry.chemical_compound ,Endocrinology ,1-Methyl-3-isobutylxanthine ,Internal medicine ,Gene expression ,Cyclic AMP ,medicine ,Animals ,Guanine Nucleotide Exchange Factors ,RNA, Messenger ,Phosphorylation ,Extracellular Signal-Regulated MAP Kinases ,Promoter Regions, Genetic ,Protein kinase A ,Forskolin ,Colforsin ,Cholera toxin ,Cyclic AMP-Dependent Protein Kinases ,Gene Expression Regulation ,chemistry ,Signal transduction ,Carrier Proteins - Abstract
Although proglucagon gene expression and the synthesis of proglucagon encoded peptide hormones could be activated by protein kinase A (PKA) activators such as forskolin/3-isobutyl-1-methylxanthine (IBMX) and cholera toxin, whether the activation is entirely attributed to PKA has not been previously examined. We found that forskolin/IBMX also activate ERK1/2 phosphorylation in intestinal and pancreatic proglucagon-producing cell lines. The MEK inhibitors PD98059 and U0126 were found to repress the expression of proglucagon promoter as well as endogenous proglucagon mRNA in two intestinal proglucagon-producing cell lines and to block the stimulatory effect of forskolin/IBMX on proglucagon mRNA expression. The repressive effect of the PKA-specific inhibitors H-89 and KT-5720, however, was either not observable or much less potent. Forskolin could activate ERK1/2 phosphorylation and proglucagon gene transcription on its own, whereas forskolin plus IBMX are required to effectively activate the PKA pathway in the proglucagon-producing cells. Exchange protein directly activated by cyclic AMP 2 (Epac2, or cAMP-binding guanine nucleotide exchange factor-2) was found to be expressed in gut and pancreatic proglucagon-producing cell lines, whereas the Epac-pathway-specific cAMP analog, 8-pMeOPT-2'O-Me-cAMP, effectively stimulated ERK1/2 phosphorylation as well as proglucagon mRNA expression. We therefore suggest that cAMP at least partially regulates proglucagon gene expression via the Epac-Ras/Rap-Raf-MEK-ERK signaling pathway.
- Published
- 2006
33. High-throughput, deterministic single cell trapping and long-term clonal cell culture in microfluidic devices
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Huaying Chen, Ernst J. Wolvetang, Jane Sun, and Justin J. Cooper-White
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Materials science ,Microfluidics ,Biomedical Engineering ,Bioengineering ,Nanotechnology ,Trapping ,Cell Separation ,Biochemistry ,Clonal analysis ,Cell size ,Lab-On-A-Chip Devices ,Cell trapping ,Humans ,Computer Simulation ,Throughput (business) ,Cells, Cultured ,Cell Proliferation ,Skin ,business.industry ,Cell growth ,General Chemistry ,Equipment Design ,Fibroblasts ,Clone Cells ,High-Throughput Screening Assays ,Cell culture ,Optoelectronics ,Single-Cell Analysis ,business - Abstract
We report the design and validation of a two-layered microfluidic device platform for single cell capture, culture and clonal expansion. Under manual injection of a cell suspension, deterministic trapping of hundreds to thousands of single cells (adherent and non-adherent) in a high throughput manner and at high trapping efficiency was achieved simply through the incorporation of a U-shaped hydrodynamic trap into the downstream wall of each micro-well. Post single cell trapping, we confirmed that these modified micro-wells permit the attachment, spreading and proliferation of the trapped single cells for multiple generations over extended periods of time (>7 days) under media perfusion. Due to its a) low cost, b) simplicity in fabrication and operation, c) high trapping efficiency, d) reliable and repeatable trapping mechanism, e) cell size selection and f) capability to provide perfused long-term culture and continuous time-lapse imaging, the microfluidic device developed and validated in this study is seen to have significant potential application in high-throughput single cell quality assessment and clonal analysis.
- Published
- 2014
34. Induction of apoptosis in MCF-7 human breast cancer cells by phytochemicals fromAnoectochilus formosanus
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Lie-Fen Shyur, Chih-Huai Chen, Chiu-Ping Lo, Sheng-Yang Wang, Pei-Ling Kang, Show-Jane Sun, C. Allen Chang, Chi-Meng Tzeng, and Ning-Sun Yang
- Subjects
Endocrinology, Diabetes and Metabolism ,Biochemistry (medical) ,Clinical Biochemistry ,Pharmacology (medical) ,Cell Biology ,General Medicine ,Molecular Biology - Published
- 2004
35. Effects of dietary lectins on ion transport in epithelia
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Jane Sun, Jens König, Rainer Schreiber, and Karl Kunzelmann
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Pharmacology ,Epithelial sodium channel ,chemistry.chemical_compound ,Gastrointestinal tract ,Phospholipase C ,chemistry ,Biochemistry ,Phytohemagglutinins ,Ussing chamber ,Mannose ,Biology ,Ion transporter ,Protein kinase C - Abstract
Phytohemagglutinins are widely distributed in common food items. They constitute a heterogeneous group of proteins, which are often resistant to proteolysis in the gastrointestinal tract. Upon binding to the luminal membrane of intestinal cells, they can interfere with digestive, protective or secretory functions of the intestine. Phytohemagglutinins present in red kidney beans and jackbeans have been shown to induce diarrhea and hypersecretion in human airways, but the underlying mechanisms remain obscure. We examined how agglutinins from wheat germ (WGA), soy bean (SBA), red kidney beans (Pha-E, Pha-L), and jackbeans (Con-A) affect ion transport in mouse airways and large intestine using Ussing chamber techniques. We found that Pha-E, Pha-L, and Con-A but not WGA and SBA inhibit electrogenic Na(+) absorption dose dependently in both colon and trachea. The inhibitory effects of Con-A on Na(+) absorption were suppressed by the sugar mannose, by inhibition of phospholipase C (PLC) and protein kinase C (PKC). Thus, nutritional phytohemagglutinins block salt absorption in a PLC- and PKC-dependent manner, probably by inhibition of the epithelial Na(+) channel (ENaC). This effect may be therapeutically useful in patients suffering from cystic fibrosis.
- Published
- 2004
36. Control of Epithelial Ion Transport by Cl− and PDZ Proteins
- Author
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Jane Sun, Rainer Schreiber, Bettina Mürle, Karl Kunzelmann, and A. Boucherot
- Subjects
Epithelial sodium channel ,Sodium-Hydrogen Exchangers ,Physiology ,PDZ domain ,Biophysics ,Xenopus ,Biological Transport, Active ,Cystic Fibrosis Transmembrane Conductance Regulator ,Sodium Channels ,Membrane Potentials ,Mice ,Animals ,Homeostasis ,Secretion ,Kidney Tubules, Collecting ,Epithelial Sodium Channels ,Cells, Cultured ,Ion transporter ,biology ,urogenital system ,Chemistry ,Sodium ,Cell Biology ,respiratory system ,Phosphoproteins ,biology.organism_classification ,Cystic fibrosis transmembrane conductance regulator ,Cell biology ,Cytosol ,biology.protein ,Chlorine ,Carrier Proteins ,Ion Channel Gating ,Intracellular - Abstract
Inhibition of epithelial Na+ channels (ENaC) by the cystic fibrosis transmembrane conductance regulator (CFTR) has been demonstrated previously. Recent studies suggested a role of cytosolic Cl- for the interaction of CFTR with ENaC, when studied in Xenopus oocytes. In the present study we demonstrate that the Na+ / H+ -exchanger regulator factor (NHERF) controls expression of CFTR in mouse collecting duct cells. Inhibition of NHERF largely attenuates CFTR expression, which is paralleled by enhanced Ca(2+) -dependent Cl- secretion and augmented Na+ absorption by the ENaC. It is further demonstrated that epithelial Na+ absorption and ENaC are inhibited by cytosolic Cl- and that stimulation by secretagogues enhances the intracellular Cl- concentration. Thus, the data provide a clue to the question, how epithelial cells can operate as both absorptive and secretory units: Increase in intracellular Cl- during activation of secretion will inhibit ENaC and switch epithelial transport from salt absorption to Cl- secretion.
- Published
- 2004
37. Subject Index Vol. 11, 2004
- Author
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Lee-Young Chau, Nan-Chi A. Chang, Irene Oi-Lin Ng, Li-Ling Chiu, Sheau-Fen Lee, Xiangyang Gong, Wen-Gang Chou, Marcelle Carolina Colhone, Chin-Chen Chu, Chiu-Hui Huang, Tanuja Singh, Nian-Chung Yang, Kuan-Hung Lin, Tseng-Long Yang, Yung-Hsi Kao, Alice Y.W. Chang, Qun Zhou, Pei-Ling Kang, Gong-Jhe Wu, Jerry M. Farley, Hsin-Yi Ho, Yuan Yao, Sebely Pal, Els J.M. Van Damme, Robin W. Rockhold, Eagle Yi-Kung Huang, Liang-Huei Lu, T.M. Wong, Ning-Sun Yang, Chien-Huang Lin, Yen-Bin Liu, Chia-Hua Kuo, Vijay Narayanasamy, Shao Hua Chen, Hui-Ling Chen, Jianzhong Sun, Wei-Tsung Chen, Alice Chien Chang, Yen-Mei Lee, Kuo-Long Chang, Tangen Ma, Sheue-Mei Wu, Tzu-Yang Lin, Joaquim Chan-Wang Lio, Yi-Jen Hsueh, George Hsiao, Yen-Hwa Chang, M. Chen, Kang-Chuang Chu, Chen-Yang Shen, Chau-Chung Wu, John C.L. Mamo, M.C.Y. Wong, Albert M. Wu, Steve S.-L. Chen, William Wei, Dong-Yan Jin, Desmond Hunt, C.H. Cho, Chiu-Ping Lo, Chi-Meng Tzeng, Lie-Fen Shyur, Mathew J. Palakal, Fu-Chan Wei, Andrew M. Thomson, Yah-Luen Lin, Paulus S. Wang, Fur-Jiang Leu, Wen-Kwei Chen, Shinn-Chih Wu, W.H. Kwong, Chien-Chuan Wang, Shen-Kou Tsai, David Potter, Li-Man Hung, Hong Zhu, Adriana Degrossoli, Ta-Liang Chen, Ing K. Ho, Jin Wang, Tzong-Shang Yang, Li-Shaung Ai, Yuan-Teh Lee, June H. Wu, Ming-Jai Su, Yen-Hsuan Ni, Jyh-Cherng Yu, Lai-Fa Sheu, Yi Chang, S. Wu, Wing-Keung Chu, Duen-Suey Chou, Shian-ling Ding, Chih-Huai Chen, Joen Rong Sheu, Snehasis Mukhopadhyay, Xizheng Zhang, Rodney C. Baker, Shih-Wei Chou, Samuel H.H. Chan, Xiao R. Li, Hsiao-Ping Wei, Wan-Jr Syu, Willy J. Peumans, Raymond T.F. Cheung, Wagner Welber Arrais-Silva, BeFong Chen, Chi Chang, Yi-Fan Yang, Shung-Tai Ho, Tsai-Yueh Luo, Ming-Yi Shen, Mao-Hsiung Yen, Marong Fang, Emma Allister, Jyh-Lyh Juang, Selma Giorgio, Pierre Rougé, Erik Helmerhorst, Solveig G. Ericson, Pao-Luh Tao, Chun-Hsien Yu, Mei-Hwei Chang, John L. Ivy, Abulkhair M. Mamoon, Wu Zhou, Yick-Pang Ching, Show-Jane Sun, C. Allen Chang, Yu-Min Cho, Hsiu-Chuan Liang, Sheng-Yang Wang, Fang Liao, David T. Yew, Karen Man-Fong Sze, and Lok-Hi Chow
- Subjects
Index (economics) ,Endocrinology, Diabetes and Metabolism ,Biochemistry (medical) ,Clinical Biochemistry ,Statistics ,Pharmacology (medical) ,Subject (documents) ,Cell Biology ,General Medicine ,Molecular Biology ,Mathematics - Published
- 2004
38. Induction of Apoptosis in MCF-7 Human Breast Cancer Cells by Phytochemicals from Anoectochilus formosanus
- Author
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Chih-Huai Chen, Sheng-Yang Wang, Chiu-Ping Lo, Ning-Sun Yang, C. Allen Chang, Show-Jane Sun, Pei-Ling Kang, Lie-Fen Shyur, and Chi-Meng Tzeng
- Subjects
medicine.diagnostic_test ,Cell growth ,Endocrinology, Diabetes and Metabolism ,Biochemistry (medical) ,Clinical Biochemistry ,Cell Biology ,General Medicine ,Biology ,Molecular biology ,Fas ligand ,Flow cytometry ,Blot ,Western blot ,Apoptosis ,Cell culture ,Cancer cell ,medicine ,Pharmacology (medical) ,Molecular Biology - Abstract
The antitumor activity of Anoectochilus formosanus Hayata (Orchidaceae), a popularly used folk medicine in the treatment of cancers in Asia, was investigated in MCF-7 human mammary carcinoma cells. Plant extracts of A. formosanus were observed to induce apoptosis of MCF-7 cells as evidenced by cell-morphological changes, an early redistribution of plasma membrane phosphatidylserine, and DNA content distribution studies. Bioactivity-guided fractionation of A. formosanus extracts produced a specific ethyl acetate (EA)-partitioned fraction in which apoptotic activity was enriched. The chemical profile and candidate index compounds of the active EA fraction were obtained using HPLC and various spectral analyses. Western blot analysis showed that upon treatment of MCF-7 cells with the EA fraction, cleavage of pro-caspases-8, -9, and -7, and poly(ADP-ribose) polymerase as well as significant release of mitochondrial cytochrome c into the cytosol were readily observed. Flow cytometry showed that the Fas ligand protein was overexpressed in EA-treated MCF-7 cells. Functional genomic studies indicated that specific genes related to cytoskeleton rearrangement, apoptotic signal transduction, and various transcription factors were differentially regulated in EA-treated MCF-7 cells. Putative apoptotic signaling pathways of MCF-7 cells in response to the EA extract of A. formosanus are proposed.
- Published
- 2004
39. Effects of Purinergic Stimulation, CFTR and Osmotic Stress on Amiloride-sensitive Na + Transport in Epithelia and Xenopus Oocytes
- Author
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Daniel Markovich, Jens König, Karl Kunzelmann, Jane Sun, and Rainer Schreiber
- Subjects
Epithelial sodium channel ,IBMX ,Physiology ,Indomethacin ,Biophysics ,Cystic Fibrosis Transmembrane Conductance Regulator ,Uridine Triphosphate ,Respiratory Mucosa ,In Vitro Techniques ,Sodium Chloride ,Biology ,Membrane Potentials ,Amiloride ,Mice ,Xenopus laevis ,chemistry.chemical_compound ,Osmotic Pressure ,1-Methyl-3-isobutylxanthine ,medicine ,Animals ,Mannitol ,Intestinal Mucosa ,Ion transporter ,Ion Transport ,Forskolin ,Colforsin ,Sodium ,Purinergic receptor ,Receptors, Purinergic ,Cell Biology ,respiratory system ,Cystic fibrosis transmembrane conductance regulator ,Hypertonic saline ,Cell biology ,Trachea ,chemistry ,Purines ,Oocytes ,biology.protein ,Female ,Ion Channel Gating ,medicine.drug - Abstract
Both stimulation of purinergic receptors by ATP and activation of the cystic fibrosis transmembrane conductance regulator (CFTR) inhibit amiloride-sensitive Na+ transport and activate Cl- secretion. These changes in ion transport may well affect cell volume. We therefore examined whether cell shrinkage or cell swelling do affect amiloride-sensitive Na+ transport in epithelial tissues or Xenopus oocytes and whether osmotic stress interferes with regulation of Na+ transport by ATP or CFTR. Stimulation of purinergic receptors by ATP/UTP or activation of CFTR by IBMX and forskolin inhibited amiloride-sensitive transport in mouse trachea and colon, respectively, by a mechanism that was Cl- dependent. When exposed to a hypertonic but not hypotonic bath solution, amiloride-sensitive Na+ transport was inhibited in mouse trachea and colon, independent of the extracellular Cl- concentration. Both inhibition of Na+ transport by hypertonic bath solution and ATP were additive. When coexpressed in Xenopus oocytes, activation of CFTR by IBMX and forskolin inhibited the epithelial Na+ channel (ENaC) in a Cl- dependent fashion. However, both hypertonic and hypotonic bath solutions showed only minor effects on amiloride-sensitive conductance, independent of the bath Cl- concentration. Moreover, CFTR-induced inhibition of ENaC could be detected in oocytes even after exposure to hypertonic or hypotonic bath solutions. We conclude that amiloride-sensitive Na+ absorption in mouse airways and colon is inhibited by cell shrinkage by a mechanism that does not interfere with purinergic and CFTR-mediated inhibition of ENaC.
- Published
- 2003
40. Deciphering the gene regulatory network necessary for head formation
- Author
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Chi Kin Ip, Joshua B. Studdert, Mirana Ramialison, Emilie E. Wilkie, Tennille Sibbritt, Joanne Shen, Nicolas Fossat, Renée Rawson, Patrick P.L. Tam, Stuart K. Archer, Vanessa Jones, and Jane Sun
- Subjects
Embryology ,Head (linguistics) ,Gene regulatory network ,Biology ,Neuroscience ,Developmental Biology - Published
- 2017
41. Mental health literacy: A focus on daily life context for population health measurement
- Author
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Kia-Chong Chua, Jane Sungmin Hahn, Suzanne Farrell, Anita Jolly, Randip Khangura, and Claire Henderson
- Subjects
Mental healing ,RZ400-408 ,Public aspects of medicine ,RA1-1270 - Published
- 2022
- Full Text
- View/download PDF
42. The Goodpasture Autoantigen
- Author
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Eric G. Neilson, Mae Jane Sun, Billy G. Hudson, and Raghu Kalluri
- Subjects
biology ,Chemistry ,Cell Biology ,Random hexamer ,medicine.disease ,Biochemistry ,Epitope ,Type IV collagen ,medicine ,biology.protein ,Goodpasture syndrome ,Binding site ,Antibody ,Molecular Biology ,Protein secondary structure ,Triple helix - Abstract
The family of type IV collagen comprises six chains numbered α1 through α6. The α3(IV) NC1 domain is the primary target antigen for autoantibodies from patients with anti-basement membrane disease and Goodpasture syndrome. Earlier peptide studies suggested that the last 36 amino acids of the α3 NC1 domain probably contains one recognition site for Goodpasture autoantibodies, and an algorithm analysis of secondary structure from a later study predicted a second possible upstream epitope near the triple helix junction. We have used several analytic approaches to evaluate the likelihood of two immunologic epitopes for the Goodpasture antigen. In our first set of studies, peptide antibodies directed against these two putative regions co-inhibited Goodpasture autoantibodies binding to denatured human α3(IV) NC1 monomer by nearly 80%, with the helix-junction region of the α3 NC1 domain contributing 26% of the binding sites and the C-terminal region contributing the remaining 50%. Second, both of these candidate regions are normally sequestered within the associated α3(IV) NC1 hexamer but become more visible for binding by anti-peptide antibodies upon their dissociation, a property that is shared by the Goodpasture autoantibodies. Third, segment deletions of recombinant α3 NC1 domain further confirmed the presence of two serologic binding sites. Finally, we looked more closely at the C-terminal binding region of the α3(IV) NC1 domain. Since the lysines in that region have been previously advanced as possible contact sites, we created several substitutions within the C-terminal epitope of the α3 NC1 domain. Substitution of lysines to alanines revealed lysines 219 and 229 as essential for antibody binding to this distal site; no lysines were present in the NC1 part of the helix-NC1 junction region. Substitutions involving arginine and cysteines to alanines in the same C-terminal region did not produce significant reductions in antibody binding. In summary, our findings characterize two Goodpasture epitopes confined to each end of the α3 NC1 domain; one is lysine-dependent, and the other is not. We propose, as a hypothetical model, that these two immunologically privileged regions fold to form an optimal pathogenic structure within the NC1 domain of the α3 chain. These sites are subsequently concealed by NC1 hexamer assembly of type IV collagen.
- Published
- 1996
43. Contents Vol. 11, 2004
- Author
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Sheau-Fen Lee, Andrew M. Thomson, Yah-Luen Lin, Xizheng Zhang, Paulus S. Wang, Fur-Jiang Leu, Shao Hua Chen, Kang-Chuang Chu, C.H. Cho, Yen-Bin Liu, Chia-Hua Kuo, Chiu-Ping Lo, Gong-Jhe Wu, Eagle Yi-Kung Huang, Sebely Pal, Els J.M. Van Damme, Liang-Huei Lu, T.M. Wong, Lai-Fa Sheu, Yi Chang, Karen Man-Fong Sze, Chau-Chung Wu, Lok-Hi Chow, Hsiao-Ping Wei, Albert M. Wu, W.H. Kwong, John L. Ivy, Abulkhair M. Mamoon, Willy J. Peumans, Chiu-Hui Huang, Tanuja Singh, Nian-Chung Yang, S. Wu, Lee-Young Chau, Tsai-Yueh Luo, Robin W. Rockhold, Ta-Liang Chen, Wan-Jr Syu, Jyh-Cherng Yu, Yick-Pang Ching, Yi-Jen Hsueh, Chien-Huang Lin, Desmond Hunt, Wei-Tsung Chen, Chien-Chuan Wang, Raymond T.F. Cheung, Alice Chien Chang, Shian-ling Ding, Fang Liao, Wing-Keung Chu, William Wei, John C.L. Mamo, Wagner Welber Arrais-Silva, BeFong Chen, Jianzhong Sun, Mao-Hsiung Yen, Chin-Chen Chu, Nan-Chi A. Chang, Yen-Hwa Chang, Joen Rong Sheu, David T. Yew, Ming-Yi Shen, Wen-Gang Chou, M.C.Y. Wong, Duen-Suey Chou, Snehasis Mukhopadhyay, Ing K. Ho, Jerry M. Farley, Hsin-Yi Ho, Adriana Degrossoli, Chih-Huai Chen, Kuan-Hung Lin, Alice Y.W. Chang, Xiangyang Gong, Marong Fang, Chi Chang, Vijay Narayanasamy, Hong Zhu, Qun Zhou, Pei-Ling Kang, Shinn-Chih Wu, Yuan Yao, Irene Oi-Lin Ng, Yi-Fan Yang, Shen-Kou Tsai, Emma Allister, Shih-Wei Chou, Yung-Hsi Kao, Yuan-Teh Lee, Shung-Tai Ho, Li-Man Hung, Tzu-Yang Lin, Ming-Jai Su, Li-Ling Chiu, June H. Wu, Show-Jane Sun, Tangen Ma, C. Allen Chang, Yu-Min Cho, Fu-Chan Wei, Tseng-Long Yang, Wen-Kwei Chen, Sheng-Yang Wang, Ning-Sun Yang, George Hsiao, Yen-Mei Lee, Kuo-Long Chang, Jyh-Lyh Juang, Sheue-Mei Wu, Chun-Hsien Yu, Selma Giorgio, Chen-Yang Shen, Yen-Hsuan Ni, Chi-Meng Tzeng, Mei-Hwei Chang, Hsiu-Chuan Liang, Rodney C. Baker, Jin Wang, Samuel H.H. Chan, Pierre Rougé, Erik Helmerhorst, Solveig G. Ericson, Mathew J. Palakal, Xiao R. Li, Pao-Luh Tao, David Potter, Wu Zhou, Steve S.-L. Chen, Tzong-Shang Yang, Li-Shaung Ai, Dong-Yan Jin, Joaquim Chan-Wang Lio, Hui-Ling Chen, M. Chen, Lie-Fen Shyur, and Marcelle Carolina Colhone
- Subjects
Endocrinology, Diabetes and Metabolism ,Biochemistry (medical) ,Clinical Biochemistry ,Pharmacology (medical) ,Cell Biology ,General Medicine ,Molecular Biology - Published
- 2004
44. Comparative distribution of the alpha 1(IV), alpha 5(IV), and alpha 6(IV) collagen chains in normal human adult and fetal tissues and in kidneys from X-linked Alport syndrome patients
- Author
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Kazuo Yoshioka, Helmut G. Rennke, Mae Jane Sun, Raghuram Kalluri, Clifford E. Kashtan, Billy G. Hudson, Eric G. Neilson, Bernard Peissel, Gloria Gallo, Jing Zhou, and Lin Geng
- Subjects
Adult ,Male ,X Chromosome ,Genetic Linkage ,Glomerulonephritis, Membranoproliferative ,Molecular Sequence Data ,Nephritis, Hereditary ,Kidney ,Basement Membrane ,law.invention ,Fetus ,Pregnancy ,law ,medicine ,Animals ,Humans ,Diabetic Nephropathies ,Amino Acid Sequence ,Alport syndrome ,Lung ,Peptide sequence ,X chromosome ,Skin ,Basement membrane ,biology ,Glomerular basement membrane ,General Medicine ,medicine.disease ,Molecular biology ,medicine.anatomical_structure ,Immunology ,biology.protein ,Recombinant DNA ,Female ,Collagen ,Rabbits ,Antibody ,Research Article - Abstract
We have shown previously that the 5' ends of the genes for the alpha 5(IV) and alpha 6(IV) collagen chains lie head-to-head on Xq22 and are deleted in patients with Alport syndrome (AS)-associated diffuse leiomyomatosis. In this study, we raised a rabbit anti-human alpha 6(IV)chain antibody, demonstrated its specificity by the analysis of recombinant NC1 domains af all six type IV chains, and studied the distribution of the alpha 6(IV) chain in relation to the alpha 1(IV) and alpha 5(IV) chains in human adult and fetal tissues involved in AS and diffuse leiomyomatosis. The alpha 6(IV) chain colocalizes with the alpha 5(IV) chain in basement membranes (BMs) of many tissues, but not in glomerular BM. These data exclude the alpha 6(IV) chain as a site for AS mutations. The head-to-head genomic pairing of the alpha 5(IV) and alpha 6 (IV) genes implies coordinate transcription of the two genes. Differential localization of the alpha 5(IV) and alpha 6(IV) chains shows that the two chains are not always coordinately regulated. The alpha 6(IV) chain, together with the alpha 3(IV)-alpha 5(IV) chains, was absent from all renal BMs in eight patients with X-linked AS while the alpha 1(IV) and alpha 2(IV) chains were increased. The data support the existence of two independent collagen networks, one for the alpha 3(IV)-alpha 6(IV) chains and one for the alpha 1(IV) and alpha 2(IV) chains.
- Published
- 1995
45. Characterization of a cis-acting regulatory element which silences expression of the class II-A beta gene in epithelium
- Author
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Mae Jane Sun, Shelley E. Albert, Frank Strutz, Thomas P. Haverty, Shao Ran Li, Kathleen Shelton, Richard A. Maki, Eric G. Neilson, and Amy Denham
- Subjects
Transcription, Genetic ,Genes, MHC Class II ,Molecular Sequence Data ,Immunology ,Response element ,Antigen presentation ,Major histocompatibility complex ,Epithelium ,Cell Line ,Mice ,Genes, Regulator ,Animals ,Immunology and Allergy ,Gene silencing ,RNA, Messenger ,Promoter Regions, Genetic ,Regulation of gene expression ,MHC class II ,Binding Sites ,Base Sequence ,biology ,Articles ,Transfection ,Molecular biology ,Kidney Tubules ,Gene Expression Regulation ,Cell culture ,biology.protein - Abstract
Class II major histocompatibility complex (MHC) genes encode for alpha/beta chain pairs that are constitutively expressed principally on mature B cells and dendritic cells in mice. These gene products are easily induced on macrophages with cytokines, and may also aberrantly appear on the surface of epithelium during immune injury. The appearance of class II determinants in parenchymal tissue potentially renders these somatic cells capable of antigen presentation to circulating CD4+ T lymphocytes, and their absence may be protective for normal tissues expressing self-antigens. The low surface class II expression observed on parenchymal cells generally correlates with low levels of mRNA, suggesting that transcription rate is a major element in class II regulation. To understand the transcriptional mechanism maintaining low basal surface expression of class II in somatic cells, we transiently transfected mini-gene reporter constructs to study the regulation of the murine A beta promoter in a cultured renal epithelial cell line. We describe here a negative cis-acting regulatory region located between -552 and -489 bp upstream of the A beta cap site that silences the transcriptional activity of the A beta promoter in epithelial cells in an orientation-dependent manner, and is also able to silence a heterologous promoter. This region is not active in class II-expressing B cells (BAL-17) in culture, but is functional in two other murine class II-negative cell lines, fibroblasts and thymoma T cells. Using competition electrophoretic mobility shift assays, we have localized the core protein binding site within this region to an 8-10-bp response element, designated A beta NRE, at -543 to -534 bp. A nuclear extract from BAL-17 cells does not bind to this element. Mutation of this site abrogates the transcriptional silencing activity of the region. We conclude that the transcription of class II-A beta in parenchymal cells, and some lymphocytes, can be actively repressed by an upstream silencing element.
- Published
- 1994
46. COL4A5 gene deletion and production of post-transplant anti-α3(IV) collagen alloantibodies in Alport syndrome
- Author
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Raghuram Kalluri, Eric G. Neilson, Kai Olaf Netzer, Manfred Weber, Mae Jane Sun, and Billy G. Hudson
- Subjects
Kidney Glomerulus ,Enzyme-Linked Immunosorbent Assay ,Nephritis, Hereditary ,Biology ,urologic and male genital diseases ,Basement Membrane ,Immunophenotyping ,Pathogenesis ,Type IV collagen ,Glomerulonephritis ,Isoantibodies ,otorhinolaryngologic diseases ,medicine ,Humans ,Alport syndrome ,Basement membrane ,Point mutation ,medicine.disease ,Kidney Transplantation ,Molecular biology ,female genital diseases and pregnancy complications ,Transplantation ,medicine.anatomical_structure ,Nephrology ,Immunology ,Electrophoresis, Polyacrylamide Gel ,Collagen ,Nephritis ,Gene Deletion - Abstract
Mutations in the COL4A5 gene encoding the alpha 5(IV) chain of type IV collagen have been implicated as the primary defect in X-linked Alport syndrome. Several kinds of mutations have been reported so far, spanning point mutations to complete gene deletions. About 5% of Alport patients, who undergo renal transplantation, develop anti-glomerular basement membrane (GBM) nephritis, causing loss of allograft function. In one such patient, COL4A5 gene deletion was recently identified. In the present study, the GBM constituent, targeted by the anti-GBM alloantibodies from the patient who had complete COL4A5 gene deletion was identified. Its identity was determined on the basis of circulating antibody binding to various GBM constituents, domains of bovine type IV collagen and recombinant NC1 domain of human type IV collagen. These results establish, for the first time, the absence of the alpha 5(IV) chain in Alport GBM and, in the same patient, the production of an alloantibody that is targeted to a different chain of type IV collagen, the alpha 3(IV) chain. These findings provide further support for the hypothesis that: (1) anti-alpha 3(IV) collagen alloantibodies mediate the allograft glomerulonephritis; and (2) COL4A5 gene mutations cause defective assembly of the alpha 3(IV) collagen alloantibodies mediate the allograft glomerulonephritis; and (2) COL4A5 gene mutations cause defective assembly of the alpha 3(IV) chain in Alport GBM, as reflected by the production of anti-alpha 3(IV) alloantibodies.
- Published
- 1994
47. Specificity of Goodpasture autoantibodies for the recombinant noncollagenous domains of human type IV collagen
- Author
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Sripad Gunwar, Eric G. Neilson, Raghuram Kalluri, Stephen T. Reeders, Jeanne C. Myers, Theodore M. Danoff, Mariko Mariyama, Mae Jane Sun, and Billy G. Hudson
- Subjects
Antiserum ,Chemistry ,Cell Biology ,medicine.disease ,Biochemistry ,Fusion protein ,Molecular biology ,Epitope ,law.invention ,Type IV collagen ,Affinity chromatography ,law ,Immunoblot Analysis ,medicine ,Recombinant DNA ,Goodpasture syndrome ,Molecular Biology - Abstract
Type IV collagen has recently emerged as a family composed of five known chains (alpha 1-alpha 5), each of which contains a carboxyl-terminal noncollagenous domain (NC1) of approximately 230 amino acids. The NC1 domain of the alpha 3(IV) chain is the probable target for autoantibodies in patients with Goodpasture syndrome (GP), as evidenced from studies employing bovine type IV collagen. In the present experiments, the specificity of GP antibodies for the five NC1 domains of human type IV collagen was determined by using recombinant NC1 domains as the antigen. cDNAs encoding each NC1 domain were expressed in E. coli as fusion proteins with a 6-histidine amino-terminal leader. The recombinant NC1 monomers r alpha 1(IV), r alpha 2(IV), r alpha 3(IV), r alpha 4(IV), and r alpha 5(IV) were purified by affinity chromatography to the fusion protein using a nickel resin column, and then characterized by electrophoresis and immunoblot analysis using chain-specific peptide antibodies. The specificity of GP antibodies from four patients to these recombinant proteins was then further evaluated by immunoblot analysis and enzyme-linked immunosorbent assay measurements. The GP antibodies reacted strongly with the r alpha 3(IV) NC1 domain but were not reactive when tested against the other four recombinant monomers. In contrast, neither antisera from patients with two other forms of autoimmune disease (anti-tubular basement membrane disease and Wegener's syndrome) nor normal control sera bound to any of the recombinant NC1 moieties. These results unambiguously establish that GP antibodies are specifically targeted to the NC1 domain of the alpha 3(IV) chain of human type IV collagen. The findings also establish a methodology for large scale preparation of r alpha 3(IV) NC1 domain for use in diagnostic tests and development of therapeutic procedures and offer a strategy for the elucidation of a more complete GP epitope by site-directed mutagenesis.
- Published
- 1993
48. Human cDNA clones transcribed from an unusually high-molecular-weight RNA encode a new collagen chain
- Author
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Jeanne C. Myers, Mae Jane Sun, Joseph A. D'Ippolito, Arnold S. Dion, Ethylin Wang Jabs, and Eric G. Neilson
- Subjects
Base Sequence ,Sequence Homology, Amino Acid ,cDNA library ,C-terminus ,Molecular Sequence Data ,Protein primary structure ,Chromosome Mapping ,RNA ,macromolecular substances ,General Medicine ,Biology ,Molecular biology ,Protein Structure, Secondary ,Molecular Weight ,Open reading frame ,Complementary DNA ,Genetics ,Humans ,Chromosomes, Human, Pair 6 ,Genomic library ,Amino Acid Sequence ,Collagen ,Cloning, Molecular ,Gene ,Gene Library - Abstract
Human collagen (COL) cDNA clones were isolated from a library representing transcripts synthesized by an established rhabdomyosarcoma (RH) cell line. The 0.6-kb insert of the first isolate encodes a discontinuous collagenous sequence not homologous to type I-XVI COL chains. Sequencing of a second clone with a 4-kb insert revealed an open reading frame (ORF) of 2154 nucleotides. The deduced amino acid (aa) sequence begins with an 186-aa noncollagenous region containing seven cysteines (Cys). Several of the Cys and surrounding aa residues can be aligned with those in type XVI, XII and IX COL. Due to the presence of two long interruptions, the 524-aa collagenous region is separated into three subdomains that each have smaller interruptions of 1-6 aa. The protein terminates with an 8-aa noncollagenous peptide including an unusual single Cys which would be expected to form an interchain disulfide bond. Results of Northern blot hybridization suggest that the new COL chain may be uncommonly large since the clone identified a low-abundance RNA at least 12.4kb in size. The gene coding for RH COL is located on human chromosome 6. It is now important to elucidate the role of this unusual COL in the infrastructure of extracellular matrix.
- Published
- 1993
49. Isolation and characterization of cDNA from renal tubular epithelium encoding murine Rantes
- Author
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Catherine Meyers, Susan O'Farrell, Mae Jane Sun, Alan M. Krensky, Gunter Wolf, Peter S. Heeger, and Eric G. Neilson
- Subjects
Chemokine ,Pathology ,medicine.medical_specialty ,Molecular Sequence Data ,Epithelium ,Kidney Tubules, Proximal ,Mice ,Paracrine signalling ,Complementary DNA ,medicine ,Animals ,Amino Acid Sequence ,Chemokine CCL5 ,Lymphokines ,Messenger RNA ,Base Sequence ,biology ,DNA ,Fusion protein ,Molecular biology ,medicine.anatomical_structure ,Nephrology ,Cell culture ,biology.protein ,Cytokines ,CD8 - Abstract
Isolation and characterization of cDNA from renal tubular epithelium encoding murine Rantes. We have been interested in identifying proinflammatory molecules which might play a role in attracting monocytes and T cells to the kidney. Some of the new intercrines are potential candidates. In this report we have isolated cDNA encoding murine Rantes (MuRantes) from renal tubular epithelium (MCT cells). MuRantes is a 91 amino acid member of the -C-C- or intercrine β subgroup of the Scy superfamily. The amino acid sequence for mature MuRantes was deduced from its coding cDNA and was found to be 90% homologous to its mature human counterpart (HuRantes). MCT epithelium expresses a single mRNA transcript for MuRantes of ∼1100 bp. The MuRantes protein could be detected in cell lysates of MCT epithelium by western blotting and in the cytoplasm of MCT cells by immunofluorescence using a polyclonal antibody generated against HuRantes fusion protein. A search protocol using MuRantes-specific primers and cDNA amplification revealed that mRNAs for MuRantes are expressed additionally in syngeneic mesangial cells (MMC cells), whole kidney, liver, and spleen, as well as in nephritogenic antigen-specific CD4 + helper and CD8 + effector T cells. cDNA amplification studies also demonstrated a significant elevation in mRNA transcripts encoding MuRantes in response to the stimulation of MCT epithelium with TNFα and IL-1α in culture, but not with TGFβ, γIFN, or IL-6. Our findings indicate that proximal tubular epithelium is an authentic source of MuRantes, and that transcripts encoding MuRantes are responsive to the modulating influence of paracrine factors having a known role in the development of parenchymal injury.
- Published
- 1992
50. POU homeodomain protein Oct-1 functions as a sensor for cyclic AMP
- Author
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Brenda B. Su, Ya-Chi Huang, Ling Liu, Hang Li, Tianru Jin, Jane Sun, Qinghua Wang, Herbert Y. Gaisano, Jing Wu, Ren-Ke Li, Jim Hu, Nina Zhang, Peixiang Wang, Harry P. Elsholtz, and Yi Zhang
- Subjects
Chromatin Immunoprecipitation ,Blotting, Western ,Molecular Sequence Data ,Biochemistry ,Cell Line ,chemistry.chemical_compound ,Islets of Langerhans ,1-Methyl-3-isobutylxanthine ,Sequence Homology, Nucleic Acid ,Cyclic AMP ,Animals ,Humans ,Nuclear Receptor Co-Repressor 1 ,CDX2 Transcription Factor ,Transcription, Chromatin, and Epigenetics ,Nuclear protein ,Promoter Regions, Genetic ,Molecular Biology ,Nuclear receptor co-repressor 1 ,Cells, Cultured ,Regulation of gene expression ,Homeodomain Proteins ,Forskolin ,biology ,Base Sequence ,Reverse Transcriptase Polymerase Chain Reaction ,Colforsin ,Nuclear Proteins ,Cell Biology ,Sequence Analysis, DNA ,Proglucagon ,Molecular biology ,Rats ,Repressor Proteins ,chemistry ,Gene Expression Regulation ,Second messenger system ,biology.protein ,Trans-Activators ,PAX4 ,RNA Interference ,Caco-2 Cells ,CREB1 ,Octamer Transcription Factor-1 ,Protein Binding - Abstract
Cyclic AMP is a fundamentally important second messenger for numerous peptide hormones and neurotransmitters that control gene expression, cell proliferation, and metabolic homeostasis. Here we show that cAMP works with the POU homeodomain protein Oct-1 to regulate gene expression in pancreatic and intestinal endocrine cells. This ubiquitously expressed transcription factor is known as a stress sensor. We found that it also functions as a repressor of Cdx-2, a proglucagon gene activator. Through a mechanism that involves the activation of exchange protein activated by cyclic AMP, elevation of cAMP leads to enhanced phosphorylation and nuclear exclusion of Oct-1 and reduced interactions between Oct-1 or nuclear co-repressors and the Cdx-2 gene promoter, detected by chromatin immunoprecipitation. In rat primary pancreatic islet cells, cAMP elevation also reduces nuclear Oct-1 content, which causes increased proglucagon and proinsulin mRNA expression. Our study therefore identifies a novel mechanism by which cAMP regulates hormone-gene expression and suggests that ubiquitously expressed Oct-1 may play a role in metabolic homeostasis by functioning as a sensor for cAMP.
- Published
- 2009
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