109 results on '"Jennifer J, Stewart"'
Search Results
2. High‐sensitivity flow cytometric assays: Considerations for design control and analytical validation for identification of Rare events
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Thomas W. McCloskey, Ulrike Sommer, Cherie Green, Jennifer J. Stewart, Teri Oldaker, Virginia Litwin, Laura Marszalek, Jolene Bradford, Alessandra Vitaliti, and Steven Eck
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0301 basic medicine ,Histology ,Computer science ,Event (computing) ,Equipment Design ,Cell Biology ,Flow Cytometry ,Design control ,Predictive value ,Lower limit ,Pathology and Forensic Medicine ,Reliability engineering ,03 medical and health sciences ,Identification (information) ,030104 developmental biology ,0302 clinical medicine ,Flow (mathematics) ,030220 oncology & carcinogenesis ,Rare events ,Humans ,Sensitivity (control systems) - Abstract
The current consensus recommendation papers dealing with the unique requirements for the analytical validation of assays performed by flow cytometry address the validation of sensitivity (both analytical and functional) only in general terms. In this paper, a detailed approach for designing and validating the sensitivity of rare event methods is described. The impact of panel design and optimization on the lower limit of quantification (LLOQ) and suggestions for reporting data near, or below, the LLOQ are addressed. This paper serves to provide best practices for the development, optimization, and analytical validation of flow cytometric assays designed to assess rare events. Note that this paper does not discuss clinical sensitivity validation, which addresses the positive and negative predictive value of the test result.
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- 2020
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3. The marine microalga, Heterosigma akashiwo, converts industrial waste gases into valuable biomass
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Jennifer J Stewart, Colleen M Bianco, Katherine R Miller, and Kathryn J Coyne
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Carbon Dioxide ,Nitric Oxide ,algae ,biofuel ,bioremediation ,biomass ,General Works - Abstract
Heterosigma akashiwo is an excellent candidate for growth on industrial emissions, since this alga has the ability to metabolize gaseous nitric oxide (NO) into cellular nitrogen via a novel chimeric protein (NR2-2/2HbN) and also tolerates wide fluctuations in temperature, salinity, and nutrient conditions. Here, we evaluated biomass productivity and composition, photosynthetic efficiency, and expression of NR2-2/2HbN for Heterosigma growing on simulated flue gas containing 12% CO2 and 150 ppm NO. Biomass productivity of Heterosigma more than doubled in flue gas conditions compared to controls, reflecting a 13-fold increase in carbohydrate and a 2-fold increase in protein productivity. Lipid productivity was not affected by flue gas and the valuable omega-3 fatty acids, EPA and DHA, constituted up to 16 % of total FAMEs. Photochemical measurements indicated that photosynthesis in Heterosigma is not inhibited by high CO2 and NO concentrations, and increases in individual fatty acids in response to flue gas were driven by photosynthetic requirements. Growth rates and maximum cell densities of Heterosigma grown on simulated flue gas without supplemental nitrogen, along with a significant increase in NR2-2/2HbN transcript abundance in response to flue gas, demonstrated that nitrogen derived from NO gas is biologically available to support enhanced CO2 fixation. Together, these results illustrate the robustness of this alga for commercial-scale biomass production and bioremediation of industrial emissions.
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- 2015
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4. The Evolution of Single-Cell Analysis and Utility in Drug Development
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Shibani Mitra-Kaushik, Anita Mehta-Damani, Cherie Green, Virginia Litwin, Christèle Gonneau, and Jennifer J. Stewart
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Microscopy ,Computer science ,flow cytometry ,Pharmacology toxicology ,Cellular biomarkers ,Pharmaceutical Science ,History, 19th Century ,Review Article ,Computational biology ,History, 20th Century ,History, 18th Century ,drug development ,History, 21st Century ,single cell ,Immune therapy ,History, 17th Century ,Drug development ,Single-cell analysis ,History, 16th Century ,Humans ,Technological advance ,Single-Cell Analysis ,Analysis tools - Abstract
This review provides a brief history of the advances of cellular analysis tools focusing on instrumentation, detection probes, and data analysis tools. The interplay of technological advancement and a deeper understanding of cellular biology are emphasized. The relevance of this topic to drug development is that the evaluation of cellular biomarkers has become a critical component of the development strategy for novel immune therapies, cell therapies, gene therapies, antiviral therapies, and vaccines. Moreover, recent technological advances in single-cell analysis are providing more robust cellular measurements and thus accelerating the advancement of novel therapies.Graphical abstract.
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- 2021
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5. Evaluation of natural killer cell assay performance on shipped blood specimens
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Mary A Fletcher, Joseph Y. Abrams, Elizabeth R. Unger, Nancy G. Klimas, Lynette Brown, Jeanne Bertolli, Troy D. Querec, Jennifer J. Stewart, Zachary Barnes, and Elizabeth Balbin
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0301 basic medicine ,Chromium ,Cytotoxicity, Immunologic ,Time Factors ,Immunology ,Pilot Projects ,Transportation ,Peripheral blood mononuclear cell ,Immunophenotyping ,Andrology ,03 medical and health sciences ,0302 clinical medicine ,Interquartile range ,Lysosomal-Associated Membrane Protein 1 ,Predictive Value of Tests ,Immunology and Allergy ,Medicine ,Humans ,Whole blood ,Cryopreservation ,Blood Specimen Collection ,business.industry ,Temperature ,Reproducibility of Results ,Gold standard (test) ,Flow Cytometry ,United States ,Killer Cells, Natural ,030104 developmental biology ,Phenotype ,Specimen collection ,Case-Control Studies ,Leukocytes, Mononuclear ,Biomarker (medicine) ,Sample collection ,business ,K562 Cells ,Biomarkers ,030215 immunology - Abstract
Documenting the importance of NK cell function as a biomarker for diseases and physiologic conditions including myalgic encephalomyelitis/chronic fatigue syndrome (ME/CFS), will require assays amenable to clinical implementation and standardization. Research studies typically perform NK functional assays on the day of sample collection. This pilot study was conducted to compare assay formats and specimen processing to identify those that are most tolerant of conditions required for shipping and amenable to standardization as shown by inter-assay and inter-laboratory correlation of results. We compared performance within and between assays that measure NK cell function using direct cytotoxicity [chromium-51 release (CRCA) or fluorescence (Flow Cytometry Cytotoxicity Assay, FCCA)] or an indirect surrogate marker (CD107a surface expression)]. Additional variables for within/between assay comparisons included time of testing (same day as specimen collection or next day within 24 h), specimen types [whole blood or isolated peripheral blood mononuclear cells (PBMCs)], and processing method (fresh or cryopreserved). Statistical measures included number of samples tested in assay conditions (n), medians (x), interquartile range (IQR), Pearson correlation coefficient (R2), and correlation p-value (p). Samples came from 3 clinics and included 31 participants. Same day testing was only available for the subset of participants enrolled from the site of the laboratory performing CRCA. Results from same day CRCA testing of whole blood were considered the gold standard [n = 10, x=10.0%, IQR = 7.2%], and correlated well with PBMCs isolated next day [n = 26, x= 15.6%, IQR = 13.1%] [R2 = 0.59, p = 0.03]. Next day CRCA results were compromised using whole blood or frozen PBMCs. Next day FCCA cytotoxicity in PBMC [n = 30, x=34.1%, IQR = 15.5%] correlated with same day CRCA PMBC [R2 = 0.8, p = 0.001] and next day CRCA PMBC [R2 = 0.5, p
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- 2020
6. Best practices for the development, analytical validation and clinical implementation of flow cytometric methods for chimeric antigen receptor T cell analyses
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Paul C Trampont, Ghanashyam Sarikonda, Virginia Litwin, Laïla-Aïcha Hanafi, Vilma Decman, Anil Pahuja, Yongliang S Sun, Mélissa Mathieu, Michael Nathan Hedrick, Susan Reynolds, Steven Eck, Qiong Xue, Naveen Dakappagari, Jennifer J. Stewart, Shabnam Tangri, Mahwish Natalia, and Piotr L Pierog
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0301 basic medicine ,Multiple stages ,Histology ,Best practice ,T cell ,T-Lymphocytes ,Computational biology ,Immunotherapy, Adoptive ,Pathology and Forensic Medicine ,Flow cytometry ,Blood cancer ,03 medical and health sciences ,0302 clinical medicine ,Medicine ,Humans ,Receptors, Chimeric Antigen ,medicine.diagnostic_test ,business.industry ,Cell Biology ,Flow Cytometry ,Chimeric antigen receptor ,Clinical trial ,030104 developmental biology ,medicine.anatomical_structure ,030220 oncology & carcinogenesis ,Car t cells ,business - Abstract
Chimeric Antigen Receptor (CAR) T cells are recognized as efficacious therapies with demonstrated ability to produce durable responses in blood cancer patients. Regulatory approvals and acceptance of these unique therapies by patients and reimbursement agencies have led to a significant increase in the number of next generation CAR T clinical trials. Flow cytometry is a powerful tool for comprehensive profiling of individual CAR T cells at multiple stages of clinical development, from product characterization during manufacturing to longitudinal evaluation of the infused product in patients. There are unique challenges with regard to the development and validation of flow cytometric methods for CAR T cells; moreover, the assay requirements for manufacturing and clinical monitoring differ. Based on the collective experience of the authors, this recommendation paper aims to review these challenges and present approaches to address them. The discussion focuses on describing key considerations for the design, optimization, validation and implementation of flow cytometric methods during the clinical development of CAR T cell therapies.
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- 2020
7. Best practices for optimization and validation of flow cytometry-based receptor occupancy assays
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Yongliang S Sun, Nathan Standifer, Ed Hilt, David Lanham, Thomas W. McCloskey, Thomas J. McIntosh, Katharine D. Grugan, Jennifer J. Stewart, Virginia Litwin, Cherie Green, and Steve Eck
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0301 basic medicine ,Cell specific ,Histology ,Occupancy ,medicine.diagnostic_test ,Computer science ,Cell Biology ,Computational biology ,Receptors, Fc ,Flow Cytometry ,Cell surface molecules ,Pathology and Forensic Medicine ,Flow cytometry ,03 medical and health sciences ,030104 developmental biology ,0302 clinical medicine ,Drug development ,Drug Development ,Pharmaceutical Preparations ,030220 oncology & carcinogenesis ,medicine ,Cellular antigens ,Humans ,Biological Assay ,Receptor - Abstract
In the development of therapeutic compounds that bind cell surface molecules, it is critical to demonstrate the extent to which the drug engages its target. For cell-associated targets, flow cytometry is well-suited to monitor drug-to-target engagement through receptor occupancy assays (ROA). The technology allows for the identification of specific cell subsets within heterogeneous populations and the detection of nonabundant cellular antigens. There are numerous challenges in the design, development, and implementation of robust ROA. Among the most difficult challenges are situations where there is receptor modulation or when the target-antigen is expressed at low levels. When the therapeutic molecules are bi-specific and bind multiple targets, these challenges are increased. This manuscript discusses the challenges and proposes best practices for designing, optimizing, and validating ROA.
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- 2020
8. Flow cytometric method transfer: Recommendations for best practice
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Maciej Cabanski, Jennifer J. Stewart, Nithianandan Selliah, Alessandra Vitaliti, Virginia Litwin, Cherie Green, Teri Oldaker, and Steve Eck
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0301 basic medicine ,Histology ,Computer science ,Best practice ,Stability (learning theory) ,Data reliability ,Cell Biology ,Flow Cytometry ,Pathology and Forensic Medicine ,Reliability engineering ,03 medical and health sciences ,030104 developmental biology ,0302 clinical medicine ,Multiple context ,Flow (mathematics) ,Acceptance testing ,030220 oncology & carcinogenesis ,Technology transfer ,Humans ,Instrumentation (computer programming) - Abstract
As with many aspects of the validation and monitoring of flow cytometric methods, the method transfer processes and acceptance criteria described for other technologies are not fully applicable. This is due to the complexity of the highly configurable instrumentation, the complexity of cellular measurands, the lack of qualified reference materials for most assays, and limited specimen stability. There are multiple reasons for initiating a method transfer, multiple regulatory settings, and multiple context of use. All of these factors influence the specific requirements for the method transfer. This recommendation paper describes the considerations and best practices for the transfer of flow cytometric methods and provides individual case studies as examples. In addition, the manuscript emphasizes the importance of appropriately conducting a method transfer on data reliability.
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- 2020
9. Single-Cell Analysis of Cytokine mRNA and Protein Expression by Flow Cytometry
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Darcey Clark, Jayne Schaubhut, Lynette Brown, Jennifer J. Stewart, and Rubina Pal
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0301 basic medicine ,Data Analysis ,Proteomics ,Histology ,Cell Membrane Permeability ,T cell ,T-Lymphocytes ,Biochemistry ,Peripheral blood mononuclear cell ,Antibodies ,Flow cytometry ,03 medical and health sciences ,0302 clinical medicine ,Single-cell analysis ,Gene expression ,medicine ,Protein biosynthesis ,Humans ,RNA, Messenger ,Messenger RNA ,medicine.diagnostic_test ,Staining and Labeling ,Chemistry ,General Medicine ,Flow Cytometry ,Molecular biology ,Medical Laboratory Technology ,030104 developmental biology ,medicine.anatomical_structure ,Cytokines ,Single-Cell Analysis ,CD8 ,030215 immunology - Abstract
Understanding how immune cells respond to external stimuli such as pathogens or drugs is a key component of biomedical research. Critical to the immune response are the expression of cell-surface receptors and the secretion of cytokines, which are tightly regulated by gene expression and protein synthesis. Previously, cytokine mRNA expression levels have been measured from bulk analysis of heterogeneous or sorted cell populations, and the correlation between cytokine mRNA expression and protein levels using these techniques can be highly variable. Flow cytometry is used to monitor changes in cell-surface and intracellular proteins, but some proteins such as cytokines may be transient and difficult to measure. Thus, a flow cytometry method that can simultaneously measure cytokine mRNA and protein levels in single cells is a very powerful tool. We defined a flow cytometry method that combines the conventional measurement of T cell surface proteins (CD45, CD3, CD4, CD8) and intracellular cytokines (IL-2, INF-γ) with fluorescent in situ hybridization and branched DNA technology for amplification and detection of IL-2 and INF-γ mRNA transcripts in activated T cells. This method has been applied to frozen peripheral mononuclear blood cells (PBMCs) and frozen blood samples, making it applicable to clinical trial specimens that require shipment to the test site. In CD4+ cells from activated PBMCs, the concordance between mRNA and protein levels was 41% for IL-2 and 21% for and INF-γ. In CD8+ cells from activated PBMCs, the concordance was 15% for IL-2 and 32% for INF-γ. © 2020 by John Wiley & Sons, Inc. Basic Protocol: Detection of IL-2 and IFN-γ mRNA and protein expression in frozen PBMCs Alternate Protocol: Detection of IL-2 and IFN-γ mRNA and protein expression in frozen blood.
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- 2020
10. Cyt-Geist: Current and Future Challenges in Cytometry: Reports of the CYTO 2018 Conference Workshops
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Kamila Czechowska, Giacomo Vacca, Cherie Green, Ruth M Barnard, Jaroslav Icha, Michael Nathan Hedrick, Gelo Victoriano Dela Cruz, Nicole E. Paul, Judith Arcidiacono, Andrew Filby, Jakob Zimmermann, Jonni S. Moore, Steven R. Bauer, Ruben Props, Jakub Nedbal, Yongliang Sun, Soren Ulrik Sonder, Christopher Hall, Caryn van Vreden, Cláudia Bispo, Paul K. Wallace, Rachel J. Errington, Jenny Molloy, Joanne Lannigan, Frederik Hammes, Johanna Ivaska, Thomas M. Ashhurst, Frederiek-Maarten Kerckhof, Michael Lee, Hyun-Dong Chang, Christian Kukat, Attila Tárnok, Nao Nitta, Robert S. Hoffman, Gert Van Isterdael, Lina Chakrabarti, John Sharpe, Michael Weber, Raluca Niesner, Christopher Groves, Peter Rubbens, Samson Rogers, Yanli Liu, Dominic Gagnon, Alessandra Vitaliti, Radhika Rayanki, Matthias Schiemann, Lili Wang, Robert Salomon, Grace Chojnowski, Rui Gardner, Bunny Cotleur, Derek H. Jones, John T. Elliott, Betsy Ohlsson-Wilhelm, Stefan Radtke, Maciej Cabanski, Ryan R. Brinkman, Michael Gregory, Henning Ulrich, Jennifer J. Stewart, Sheng Lin-Gibson, Dominic C. Jenner, Heba A. Degheidy, Virginia Litwin, Ziv Portat, Silas J. Leavesley, David Lanham, Susann Müller, Stephen P. Perfetto, Steven Eck, Aja M. Rieger, and Diana L. Bonilla
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Histology ,Drug development ,Geist ,Immunology ,570 Life sciences ,biology ,Cell Biology ,Computational biology ,Biomarker discovery ,Biology ,500 Science ,Cytometry ,Pathology and Forensic Medicine - Published
- 2019
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11. Light intensity impacts the production of biofuel intermediates in Heterosigma akashiwo growing on simulated flue gas containing carbon dioxide and nitric oxide
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Catherine Fitzgerald, Kathryn J. Coyne, Katherine R. Miller, Jennifer J. Stewart, and Colleen M. Bianco
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0301 basic medicine ,Environmental Engineering ,Light ,Industrial Waste ,Photobioreactor ,Bioengineering ,010501 environmental sciences ,Nitric Oxide ,complex mixtures ,01 natural sciences ,7. Clean energy ,Photobioreactors ,03 medical and health sciences ,chemistry.chemical_compound ,Waste Management ,Bioenergy ,Botany ,Microalgae ,Biomass ,Food science ,Waste Management and Disposal ,0105 earth and related environmental sciences ,chemistry.chemical_classification ,Biodiesel ,biology ,Renewable Energy, Sustainability and the Environment ,Fatty acid ,General Medicine ,Carbon Dioxide ,biology.organism_classification ,Lipids ,Light intensity ,030104 developmental biology ,chemistry ,13. Climate action ,Biofuel ,Biofuels ,Carbon dioxide ,Heterosigma akashiwo - Abstract
As a potential biofuel feedstock, the marine microalga, Heterosigma akashiwo, accumulates significant lipids, is capable of long-term growth in outdoor photobioreactors, and is an excellent candidate for the bioremediation of industrial emissions. Here, we evaluated resource partitioning in H. akashiwo growing on a CO2 and NO gas mixture under three light intensities: 160, 560, or 1200 μmol quanta m−2 s−1. Light levels had no effect on growth; however, cultures in high light accumulated 2.3-fold more carbohydrates and 17% fewer lipids. Light levels did not affect the percentage of saturated fatty acids, but mono-unsaturates increased by 6% and poly-unsaturates decreased by 12% in high light. The fatty acid profiles reported here suggest that H. akashiwo is a good candidate for the production of neutral lipids for biodiesel and also omega-3 fatty acids, and that the quality of biodiesel acquired from feedstocks grown under fluctuating light conditions would be relatively stable.
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- 2016
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12. Recommendations for the development and validation of flow cytometry-based receptor occupancy assays
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Barry van der Strate, John Ferbas, Virginia Litwin, Tim Wyant, Thomas W. Mc Closkey, Jennifer J. Stewart, Cherie Green, Meina Liang, Yuanxin Xu, Nicholas Jones, Carl-Magnus Högerkorp, David Lanham, Danice E. C. Wilkins, Alan Lackey, Kamila Czechowska, and Maxime Moulard
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0301 basic medicine ,Histology ,Occupancy ,medicine.diagnostic_test ,Chemistry ,Cell Biology ,Pharmacology ,Pathology and Forensic Medicine ,Flow cytometry ,03 medical and health sciences ,030104 developmental biology ,0302 clinical medicine ,Drug development ,030220 oncology & carcinogenesis ,Pharmacodynamics ,Biotherapeutic agent ,medicine ,Receptor ,Cytometry ,Dose selection - Abstract
Receptor occupancy measurements demonstrate the binding of a biotherapeutic agent to its extra-cellular target and represent an integral component of the pharmacodynamic (PD) portfolio utilized to advance the development and commercialization of a therapeutic agent. Coupled with traditional pharmacokinetic (PK) assessments derived from serum drug concentration, receptor occupancy data can be used to model PK/PD relationships and validate dose selection decisions throughout the drug development lifecycle. Receptor occupancy assays can be even more challenging to develop than other flow cytometric methods (e.g. surface immunophenotyping). In addition to typical considerations regarding stability of the cell type of interest, stability of the target-bound therapeutic agent and stability of the target receptor must be taken into account. Reagent selection is also challenging as reagents need to be evaluated for the potential to compete with the therapeutic agent and bind with comparable affinity. This article provides technical guidance for the development and validation of cytometry-based receptor occupancy assays.
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- 2016
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13. Role of receptor occupancy assays by flow cytometry in drug development
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Tim Wyant, David Lanham, Danice E. C. Wilkins, Meina Liang, Nicholas Jones, Jennifer J. Stewart, Kamila Czechowska, Alan Lackey, Barry van der Strate, Carl-Magnus Högerkorp, John Ferbas, Yuanxin Xu, Thomas W. McCloskey, Virginia Litwin, Maxime Moulard, and Cherie Green
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0301 basic medicine ,Histology ,medicine.diagnostic_test ,Drug discovery ,media_common.quotation_subject ,Cell ,Cell Biology ,Pharmacology ,Biology ,Pathology and Forensic Medicine ,Biomarker (cell) ,Flow cytometry ,03 medical and health sciences ,030104 developmental biology ,medicine.anatomical_structure ,Drug development ,Cell surface receptor ,medicine ,Receptor ,Internalization ,media_common - Abstract
The measurement of the binding of a biotherapeutic to its cellular target, receptor occupancy (RO), is increasingly important in development of biologically-based therapeutic agents. Receptor occupancy (RO) assays by flow cytometry describe the qualitative and/or quantitative assessment of the binding of a therapeutic agent to its cell surface target. Such RO assays can be as simple as measuring the number of cell surface receptors bound by an antireceptor therapeutic agent or can be designed to address more complicated scenarios such as internalization or shedding events once a receptor engages the administered therapeutic agent. Data generated from RO assays can also be used to model whether given doses of an experimental therapeutic agent and their administration schedules lead to predicted levels of receptor occupancy and whether the receptor is modulated (up or down) on cells engaged by the therapeutic agent. There are a variety of approaches that can be used when undertaking RO assays and with the ability to measure distinct subsets in heterogeneous populations, flow cytometry is ideally suited to RO measurements. This article highlights the importance of RO assays on the flow cytometric platform in the development of biotherapeutic agents.
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- 2016
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14. A Microalgae-Based Platform for the Beneficial Re-use of Carbon Dioxide Emissions from Power Plants
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Eduardo Santillan-Jimenez, Mark Crocker, Jenna Y. Schambach, Jack Groppo, Robby Pace, Jennifer J. Stewart, Ashton Zeller, Daniel T. Mohler, Stephanie Kesner, and Michael H. Wilson
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chemistry.chemical_compound ,chemistry ,Waste management ,Carbon dioxide ,Environmental science ,Power (physics) - Published
- 2018
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15. Beneficial re-use of industrial CO2 emissions using microalgae: Demonstration assessment and biomass characterization
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Daniel T. Mohler, Mark Crocker, Stephanie Kesner, Darin Vaughan, Jenna Y. Schambach, Jack Groppo, Michael H. Wilson, Molly Frazar, Jennifer J. Stewart, and Robert Pace
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0106 biological sciences ,Environmental Engineering ,biology ,Renewable Energy, Sustainability and the Environment ,Biomass ,Photobioreactor ,Bioengineering ,Heavy metals ,General Medicine ,010501 environmental sciences ,biology.organism_classification ,01 natural sciences ,chemistry.chemical_compound ,Nutrient ,Algae ,chemistry ,Productivity (ecology) ,010608 biotechnology ,Environmental chemistry ,Carbon dioxide ,Environmental science ,Composition (visual arts) ,Waste Management and Disposal ,0105 earth and related environmental sciences - Abstract
A novel cyclic flow photobioreactor, designed for the capture and recycle of CO2 using microalgae, was deployed at a coal-fired power plant. Scenedesmus acutus was cultured continuously for a four-month period, during which a biomass productivity of 0.1–0.2 g L−1 day−1 was observed. Samples taken for DNA sequencing showed a strong correlation between the composition of the culture and environmental conditions. Dry and liquid biomass samples and the industrial fertilizers used for preparation of the nutrient medium were analyzed to determine the presence of heavy metals (As, Cd, Hg, Se) and results were compared with standardized and/or regulated maximum contaminant levels (MCLs) for metals in several possible algae derived products. Concentrations of the metals in dry algae biomass were consistent with the incorporation of metals from the supplied nutrients.
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- 2019
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16. Analysis of raphidophyte assimilatory nitrate reductase reveals unique domain architecture incorporating a 2/2 hemoglobin
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Kathryn J. Coyne and Jennifer J. Stewart
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Hemeprotein ,Nitrogen assimilation ,Nitric oxide dioxygenase ,Plant Science ,General Medicine ,Reductase ,Biology ,biology.organism_classification ,Nitrate reductase ,Protein Structure, Tertiary ,Nitric oxide ,chemistry.chemical_compound ,Nitrate ,chemistry ,Biochemistry ,Nitrate Reductases ,Rhodophyta ,Genetics ,Heterosigma akashiwo ,Agronomy and Crop Science ,Phylogeny - Abstract
Eukaryotic assimilatory nitrate reductase (NR) is a multi-domain protein that catalyzes the rate-limiting step in nitrate assimilation. This protein is highly conserved and has been extensively characterized in plants and algae. Here, we report hybrid NRs (NR2-2/2HbN) identified in two microalgal species, Heterosigma akashiwo and Chattonella subsalsa, with a 2/2 hemoglobin (2/2Hb) inserted into the hinge 2 region of a prototypical NR. 2/2Hbs are a class of single-domain heme proteins found in bacteria, ciliates, algae and plants. Sequence analysis indicates that the C-terminal FAD/NADH reductase domain of NR2-2/2HbN retains identity with eukaryotic NR, suggesting that the 2/2Hb domain was inserted interior to the existing NR domain architecture. Phylogenetic analysis supports the placement of the 2/2Hb domain of NR2-2/2HbN within group I (N-type) 2/2Hbs with high similarity to mycobacterial 2/2HbNs, known to convert nitric oxide to nitrate. Experimental data confirms that H. akashiwo is capable of metabolizing nitric oxide and shows that HaNR2-2/2HbN expression increases in response to nitric oxide addition. Here, we propose a mechanism for the dual function of NR2-2/2HbN in which nitrate reduction and nitric oxide dioxygenase reactions are cooperative, such that conversion of nitric oxide to nitrate is followed by reduction of nitrate for assimilation as cellular nitrogen.
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- 2011
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17. RBC-Specific CD47 Pruning Confers Protection and Underlies the Transient Anemia in Patients Treated with Anti-CD47 Antibody 5F9
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Chris H. Takimoto, Jiaqi Duan, Kavitha Sompalli, Mark P. Chao, Timothy S Choi, Lynette Brown, Kyle Elrod, Ravindra Majeti, Jie Liu, Jennifer J. Stewart, Jens-Peter Volkmer, Mckenna Kelly Marie, James Y. Chen, Stanley L. Schrier, Paresh Vyas, and Irving L. Weissman
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biology ,business.industry ,Reticulocytosis ,CD47 ,Immunology ,Priming (immunology) ,Context (language use) ,Cell Biology ,Hematology ,030204 cardiovascular system & hematology ,Biochemistry ,03 medical and health sciences ,0302 clinical medicine ,Immune system ,medicine.anatomical_structure ,Antigen ,030220 oncology & carcinogenesis ,biology.protein ,Medicine ,Bone marrow ,Antibody ,medicine.symptom ,business - Abstract
CD47 is an anti-phagocytic (i.e. "don't eat me") signal and macrophage checkpoint that cancer cells utilize to evade innate immunity and establish disease. 5F9 is a humanized IgG4 monoclonal antibody (mAb) that binds to human CD47 and blocks its interaction with SIRPα, its cognate inhibitory receptor.5F9 is undergoing investigation in several clinical trials and preliminary analyses have revealed encouraging therapeutic potential. We previously established that administration of 5F9 clears a subset of aged RBCs, resulting in a compensatory reticulocytosis. This occurs transiently, predominantly after the first dose, and does not worsen with subsequent doses. These findings, aligned with the observations of Oldenborg et al., which established that removal of aged RBCs, is dependent on the balance between the accumulating pro-phagocytic signals and loss of inhibitory anti-phagocytic signals. We therefore hypothesized that an initial lower 5F9 "priming dose", sufficient to trigger clearance of aged RBCs, would yield an overall younger pool of RBCs that are less vulnerable to subsequent higher 5F9 therapeutic "maintenance doses". Indeed, this priming and maintenance dosing strategy has substantially mitigated the on-target anemia and has allowed for 5F9 treatment in multiple clinical trials and indications. To further probe this observed selective RBC clearance, we employed a receptor occupancy assay to quantitate the CD47 expression on the cell surface of RBCs, leukocytes (WBCs), and acute myeloid leukemia (AML) blasts. We discovered that the priming dose of 5F9 not only triggered clearance of a subset of RBCs, but also resulted in a near complete loss of CD47 - a phenomenon we term CD47 pruning. This effect was RBC-specific, as WBCs and AML bone marrow (BM) blasts did not exhibit this same CD47 pruning (Fig 1). To explore the functional consequence of these unexpected clinical findings, we investigated whether we could recapitulate them in a preclinical mouse model. Similar to the clinical setting, anti-mouse CD47 blocking Ab treatments exhibited a similar pruning effect and transient anemia. Using this model, we asked whether, in the context of anti-CD47 treatment, these pruned RBCs were protected compared to unpruned RBCs. We transfused fluorescently labeled RBCs from CD47 Ab-untreated and CD47-treated mice into animals receiving continuous anti-CD47 treatment. We observed that CD47-pruned RBCs exhibited a significantly longer half-life compared to unpruned RBCs, suggesting that CD47 pruning is protective for RBCs. To distinguish whether this protection is cell-intrinsic or cell-extrinsic, we transfused fluorescently labeled RBCs into CD47 Ab-untreated and -treated mice and observed that CD47-pruned RBCs are rapidly cleared in untreated recipients, suggesting that this treatment-induced protection of RBCs is cell extrinsic and a host-based alteration. Through depletion of distinct immune subsets by chemical depletion, antibody depletion, splenectomy, and partial hepatectomies, we demonstrated that host immune effector cells (e.g. red-pulp macrophages, neutrophils, T cells etc.) were not necessary for this pruning phenomenon. Critically, we also found that this CD47 pruning and resulting tolerance was Fc-independent. In order to understand whether these findings were generalizable to other anti-human CD47 blocking agents, we conducted similar studies in double humanized huCD47 and huSIRPa micewith additional agents and observed this same CD47 pruning and tolerance phenomenon. Thereby suggesting this represents a class-effect of this emerging potential therapy class. Notably, these clinical and pre-clinical findings have been RBC-specific and have not precluded anti-tumor efficacy. The loss of CD47 on RBCs after the priming dose also suggests that the potential risk of CD47 Ab-mediated RBC agglutination following subsequent maintenance dosing is substantially reduced. Unlike prior RBC antigen modulation reports, to our knowledge, these findings represent a novel RBC antigen depletion phenomenon that is independent of the known RBC regulatory compartments, including the spleen, liver, major immune effectors cells, complement, and is critically Fc-independent. These findings provide fundamental insight into the mechanism underlying how anti-CD47 Abs are tolerated without impairing therapeutic efficacy. Disclosures Chen: Forty Seven Inc: Consultancy, Equity Ownership. McKenna:Forty Seven Inc.: Equity Ownership. Choi:Forty Seven Inc: Employment, Equity Ownership. Duan:Forty Seven Inc: Employment, Equity Ownership. Sompalli:Forty Seven Inc: Employment, Equity Ownership. Schrier:Forty Seven Inc.: Consultancy. Weissman:Forty Seven, Inc: Consultancy, Equity Ownership, Membership on an entity's Board of Directors or advisory committees, Patents & Royalties. Elrod:Forty Seven Inc.: Employment, Equity Ownership. Chao:Forty Seven Inc: Employment, Equity Ownership, Patents & Royalties. Takimoto:Forty Seven Inc: Employment, Equity Ownership, Patents & Royalties. Liu:Forty Seven Inc: Employment, Equity Ownership, Patents & Royalties. Volkmer:Forty Seven Inc: Employment, Equity Ownership, Patents & Royalties.
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- 2018
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18. Flow Cytometry Method Validation Protocols
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Steven Eck, Cherie Green, Virginia Litwin, Jennifer J. Stewart, Nithianandan Selliah, Teri Oldaker, and Alessandra Vitaliti
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Quality Control ,0301 basic medicine ,Protocol (science) ,Histology ,Computer science ,Reproducibility of Results ,General Medicine ,Flow Cytometry ,Biochemistry ,03 medical and health sciences ,Medical Laboratory Technology ,030104 developmental biology ,0302 clinical medicine ,Limit of Detection ,Systems engineering ,Animals ,Humans ,030215 immunology - Abstract
Analytical method validation provides a means to ensure that data are credible and reproducible. This unit will provide a brief introduction to analytical method validation as applied to cellular analysis by flow cytometry. In addition, the unit will provide practical procedures for three different types of validation. The first is a limited validation protocol that is applicable for research settings and non-regulated laboratories. The second is validation protocol that presents the minimum validation requirements in regulated laboratories. The third is a transfer validation protocol to be used when methods are transferred between laboratories. The recommendations presented in this unit are consistent with the white papers published by the American Association of Pharmaceutical Scientists and the International Clinical Cytometry Society, as well as with Clinical Laboratory Standards Institute Guideline H62: Validation of Assays Performed by Flow Cytometry (currently in preparation). © 2018 by John Wiley & Sons, Inc.
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- 2018
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19. The marine microalga, Heterosigma akashiwo, converts industrial waste gases into valuable biomass
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Katherine R. Miller, Colleen M. Bianco, Jennifer J. Stewart, and Kathryn J. Coyne
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Flue gas ,Economics and Econometrics ,chemistry.chemical_element ,Biomass ,Energy Engineering and Power Technology ,lcsh:A ,Photosynthetic efficiency ,Photosynthesis ,Nitric Oxide ,7. Clean energy ,chemistry.chemical_compound ,bioremediation ,Botany ,algae ,biology ,biomass ,Renewable Energy, Sustainability and the Environment ,Carbon Dioxide ,biology.organism_classification ,Energy Research ,Nitrogen ,Fuel Technology ,chemistry ,13. Climate action ,Biofuel ,Environmental chemistry ,Carbon dioxide ,biofuel ,raphidophyte ,lcsh:General Works ,Heterosigma akashiwo - Abstract
Heterosigma akashiwo is an excellent candidate for growth on industrial emissions since this alga has the ability to metabolize gaseous nitric oxide (NO) into cellular nitrogen via a novel chimeric protein (NR2-2/2HbN) and also tolerates wide fluctuations in temperature, salinity, and nutrient conditions. Here, we evaluated biomass productivity and composition, photosynthetic efficiency, and expression of NR2-2/2HbN for Heterosigma growing on simulated flue gas containing 12% CO2 and 150 ppm NO. Biomass productivity of Heterosigma more than doubled in flue gas conditions compared to controls, reflecting a 13-fold increase in carbohydrate and a 2-fold increase in protein productivity. Lipid productivity was not affected by flue gas and the valuable omega-3 fatty acids, eicosapentaenoic acid and docosahexaenoic acid, constituted up to 16% of total fatty acid methyl esters. Photochemical measurements indicated that photosynthesis in Heterosigma is not inhibited by high CO2 and NO concentrations, and increases in individual fatty acids in response to flue gas were driven by photosynthetic requirements. Growth rates and maximum cell densities of Heterosigma grown on simulated flue gas without supplemental nitrogen, along with a significant increase in NR2-2/2HbN transcript abundance in response to flue gas, demonstrated that nitrogen derived from NO gas is biologically available to support enhanced CO2 fixation. Together, these results illustrate the robustness of this alga for commercial-scale biomass production and bioremediation of industrial emissions.
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- 2015
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20. Proteins Associated with Cisplatin Resistance in Ovarian Cancer Cells Identified by Quantitative Proteomic Technology and Integrated with mRNA Expression Levels
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Nicole Urban, James T. White, Jennifer J. Stewart, Biaoyang Lin, Charles W. Drescher, Xiaowei Yan, Steven J. Collins, and Leroy Hood
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Proteomics ,Cell ,Protein Array Analysis ,Biochemistry ,Analytical Chemistry ,Drug Therapy ,Cell Line, Tumor ,Tumor Cells, Cultured ,medicine ,Humans ,RNA, Messenger ,Claudin ,Protein kinase A ,Molecular Biology ,Ovarian Neoplasms ,Cisplatin ,Chemistry ,Cancer ,Combination chemotherapy ,medicine.disease ,Molecular biology ,Neoplasm Proteins ,Gene Expression Regulation, Neoplastic ,medicine.anatomical_structure ,Drug Resistance, Neoplasm ,Proteome ,Female ,Ovarian cancer ,Genes, Neoplasm ,medicine.drug - Abstract
Nearly all women diagnosed with ovarian cancer receive combination chemotherapy including cis- or carboplatin. Despite high initial response rates, resistance to cisplatin develops in roughly one-third of women during primary treatment and in all women treated for recurrent disease. ICAT coupled with tandem MS is a quantitative proteomic technique for high throughput protein expression profiling of complex protein mixtures. Using ICAT/MS/MS we profiled the nuclear, cytosolic, and microsomal fractions obtained from IGROV-1 [corrected] (cisplatin-sensitive) and IGROV-1/CP [corrected] (cisplatin-resistant) ovarian cancer cell lines. The proteomes of cisplatin-sensitive and -resistant ovarian cancer cells were compared, and protein expression was correlated with mRNA expression profiles. A total of 1117 proteins were identified and quantified. The relative expression of 121 of these varied between the two cell lines. Sixty-three proteins were overexpressed in cisplatin-sensitive, and 58 were over expressed in cisplatin-resistant cells. Examples of proteins at least 5-fold overexpressed in resistant cells and with biological relevance to cancer include cell recognition molecule CASPR3 (13.3-fold), S100 protein family members (8.7-fold), junction adhesion molecule Claudin 4 (7.2-fold), and CDC42-binding protein kinase beta (5.4-fold). Examples of cancer-related proteins at least 5-fold overexpressed in sensitive cells include hepatocyte growth factor inhibitor 1B (13.3-fold) and programmed cell death 6-interacting protein (12.7-fold). The direction of changes in expression levels between proteins and mRNAs were not always in the same direction, possibly reflecting posttranscriptional control of protein expression. We identified proteins whose expression profiles correlate with cisplatin resistance in ovarian cancer cells. Several proteins may be involved in modulating response to cisplatin and have potential as markers of treatment response or treatment targets.
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- 2006
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21. Receptor occupancy by flow cytometry
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Cherie Green, Virginia Litwin, and Jennifer J. Stewart
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0301 basic medicine ,Histology ,Occupancy ,medicine.diagnostic_test ,business.industry ,Cell Biology ,Computational biology ,Pathology and Forensic Medicine ,Flow cytometry ,03 medical and health sciences ,030104 developmental biology ,Text mining ,Medicine ,business ,Receptor ,Introductory Journal Article - Published
- 2016
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22. Role of receptor occupancy assays by flow cytometry in drug development
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Jennifer J, Stewart, Cherie L, Green, Nicholas, Jones, Meina, Liang, Yuanxin, Xu, Danice E C, Wilkins, Maxime, Moulard, Kamila, Czechowska, David, Lanham, Thomas W, McCloskey, John, Ferbas, Barry W A, van der Strate, Carl-Magnus, Högerkorp, Timothy, Wyant, Alan, Lackey, and Virginia, Litwin
- Subjects
Drug Discovery ,Humans ,Flow Cytometry ,Antibodies - Abstract
The measurement of the binding of a biotherapeutic to its cellular target, receptor occupancy (RO), is increasingly important in development of biologically-based therapeutic agents. Receptor occupancy (RO) assays by flow cytometry describe the qualitative and/or quantitative assessment of the binding of a therapeutic agent to its cell surface target. Such RO assays can be as simple as measuring the number of cell surface receptors bound by an antireceptor therapeutic agent or can be designed to address more complicated scenarios such as internalization or shedding events once a receptor engages the administered therapeutic agent. Data generated from RO assays can also be used to model whether given doses of an experimental therapeutic agent and their administration schedules lead to predicted levels of receptor occupancy and whether the receptor is modulated (up or down) on cells engaged by the therapeutic agent. There are a variety of approaches that can be used when undertaking RO assays and with the ability to measure distinct subsets in heterogeneous populations, flow cytometry is ideally suited to RO measurements. This article highlights the importance of RO assays on the flow cytometric platform in the development of biotherapeutic agents.
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- 2015
23. Recommendations for the evaluation of specimen stability for flow cytometric testing during drug development
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Wendy I. White, Stephanie Fraser, Carla G. Hill, Peter J. O'Brien, Christopher A. Wiwi, Jennifer J. Stewart, Kathy Howell, Cherie Green, Yuanxin Xu, Virginia Litwin, Nicholas Jones, and Lynette Brown
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Blood Specimen Collection ,Computer science ,Immunology ,Stability (learning theory) ,Analytic Sample Preparation Methods ,Flow Cytometry ,Method development ,Stability assessment ,Reliability engineering ,Drug development ,Acceptance testing ,Drug Discovery ,Immunology and Allergy ,Humans - Abstract
The objective of this manuscript is to present an approach for evaluating specimen stability for flow cytometric methods used during drug development. While this approach specifically addresses stability assessment for assays to be used in clinical trials with centralized testing facilities, the concepts can be applied to any stability assessment for flow cytometric methods. The proposed approach is implemented during assay development and optimization, and includes suggestions for designing a stability assessment plan, data evaluation and acceptance criteria. Given that no single solution will be applicable in all scenarios, this manuscript offers the reader a roadmap for stability assessment and is intended to guide the investigator during both the method development phase and in the experimental design of the validation plan.
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- 2014
24. The Stability of the TFIIA-TBP-DNA Complex Is Dependent on the Sequence of the TATAAA Element
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Laurie A. Stargell and Jennifer J. Stewart
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Time Factors ,Transcription, Genetic ,Protein subunit ,genetic processes ,macromolecular substances ,Plasma protein binding ,Biology ,Biochemistry ,In vivo ,Transcription (biology) ,Yeasts ,Promoter Regions, Genetic ,Molecular Biology ,Repetitive Sequences, Nucleic Acid ,Genetics ,TATA-Box Binding Protein ,Promoter ,DNA ,Cell Biology ,Recombinant Proteins ,DNA-Binding Proteins ,Kinetics ,enzymes and coenzymes (carbohydrates) ,Transcription Factor TFIIA ,health occupations ,Nucleic acid ,Biophysics ,Transcription factor II A ,Plasmids ,Protein Binding ,Transcription Factors - Abstract
To determine the mechanistic differences between canonical and noncanonical TATA elements, we compared the functional activity of two sequences: TATAAA (canonical) and CATAAA (noncanonical). The TATAAA element can support high levels of transcription in vivo, whereas the CATAAA element is severely defective for this function. This dramatic functional difference is not likely to be due to a difference in TBP (TATA-binding protein) binding efficiency because protein-DNA complex studies in vitro indicate little difference between the two DNA sequences in the formation and stability of the TBP-DNA complex. In addition, the binding and stability of the TFIIB-TBP-DNA complex is similar for the two elements. In striking contrast, the TFIIA-TBP-DNA complex is significantly less stable on the CATAAA element when compared with the TATAAA element. A role for TFIIA in distinguishing between TATAAA and CATAAA in vivo was tested by fusing a subunit of TFIIA to TBP. We found that fusion of TFIIA to TBP dramatically increases transcription from CATAAA in yeast cells. Taken together, these results indicate that the stability of the TFIIA-TBP complex depends strongly on the sequence of the core promoter element and that the TFIIA-TBP complex plays an important function in recognizing optimal promoters in vivo.
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- 2001
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25. Recognition between symbiotic Vibrio fischeri and the haemocytes of Euprymna scolopes
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Edward G. Ruby, Spencer V. Nyholm, Jennifer J. Stewart, and Margaret J. McFall-Ngai
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Innate immune system ,Hemocytes ,biology ,Euprymna scolopes ,Decapodiformes ,biochemical phenomena, metabolism, and nutrition ,biology.organism_classification ,Microbiology ,Aliivibrio fischeri ,Vibrio ,Article ,Light organ ,Immune system ,Phagocytosis ,bacteria ,Animals ,Bacterial outer membrane ,Symbiosis ,Ecology, Evolution, Behavior and Systematics ,Bacteria ,Gene Deletion ,Bacterial Outer Membrane Proteins - Abstract
The light organ crypts of the squid Euprymna scolopes permit colonization exclusively by the luminous bacterium Vibrio fischeri. Because the crypt interior remains in contact with seawater, the squid must not only foster the specific symbiosis, but also continue to exclude other bacteria. Investigation of the role of the innate immune system in these processes revealed that macrophage-like haemocytes isolated from E. scolopes recognized and phagocytosed V. fischeri less than other closely related bacterial species common to the host's environment. Interestingly, phagocytes isolated from hosts that had been cured of their symbionts bound five times more V. fischeri cells than those from uncured hosts. No such change in the ability to bind other species of bacteria was observed, suggesting that the host adapts specifically to V. fischeri. Deletion of the gene encoding OmpU, the major outer membrane protein of V. fischeri, increased binding by haemocytes from uncured animals to the level observed for haemocytes from cured animals. Co-incubation with wild-type V. fischeri reduced this binding, suggesting that they produce a factor that complements the mutant's defect. Analyses of the phagocytosis of bound cells by fluorescence-activated cell sorting indicated that once binding to haemocytes had occurred, V. fischeri cells are phagocytosed as effectively as other bacteria. Thus, discrimination by this component of the squid immune system occurs at the level of haemocyte binding, and this response: (i) is modified by previous exposure to the symbiont and (ii) relies on outer membrane and/or secreted components of the symbionts. These data suggest that regulation of host haemocyte binding by the symbiont may be one of many factors that contribute to specificity in this association.
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- 2009
26. Non-optimal TATA elements exhibit diverse mechanistic consequences
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Julie A. Fischbeck, Xu Chen, Laurie A. Stargell, and Jennifer J. Stewart
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Guanine ,Transcription, Genetic ,TATA box ,Molecular Sequence Data ,Biology ,Biochemistry ,Fungal Proteins ,Cytosine ,In vivo ,Transcription (biology) ,Escherichia coli ,Binding site ,Promoter Regions, Genetic ,Molecular Biology ,Gene ,Regulation of gene expression ,Genetics ,Binding Sites ,Base Sequence ,Cell Biology ,TATA-Box Binding Protein ,TATA Box ,Recombinant Proteins ,Cell biology ,enzymes and coenzymes (carbohydrates) ,Gene Expression Regulation ,Transcription factor II B ,Transcription factor II A ,Protein Binding - Abstract
To reveal mechanistic differences in transcription initiation between variant TATA elements, in vivo and in vitro assays of the functional activity of 14 different sequences were compared. Variant elements exhibited particular degrees of activation in vivo but universally were unable to support the -fold activation observed for an element consisting of TATAAA. Each element was classified by its functional activity for in vitro interaction with TATA-binding protein (TBP), TFIIA, and TFIIB. Certain off-consensus TATA elements form poor binding sites for TBP and this compromised interaction interferes with higher order complex formation with TFIIA and/or TFIIB. Other elements are only modestly decreased for TBP binding but dramatically affected for higher order complex formation. Another distinct category is comprised of two elements (CATAAA and TATAAG), which are not affected in the initial formation of the TBP, TFIIA-TBP, or TFIIB-TBP complexes. However, CATAAA and TATAAG are unable to form a stable TFIIA-TBP-DNA complex in vitro. Moreover, fusion of TFIIA to TBP specifically restores activity from these two elements in vivo. Taken together, these results indicate that the interplay between the sequence of the TATA element and the components of the general transcription machinery can lead to variations in the formation of functional complexes and/or the stability of these complexes. These differences offer distinct opportunities for an organism to exploit diverse steps in the regulation of gene expression depending on the precise TATA element sequence at a given gene.
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- 2006
27. An annotated cDNA library of juvenile Euprymna scolopes with and without colonization by the symbiont Vibrio fischeri
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Tad DeMartini, Keith Crouch, Margaret J. McFall-Ngai, Deyan Tong, Tanya A. Koropatnick, Jennifer R. Kimbell, Jennifer J. Stewart, Todd E. Scheetz, Mari E. Eyestone, Bartley Brown, Michael S. Goodson, Tamara A. Kucaba, Bernadette Janssens, Joshua V. Troll, Thomas L. Casavant, Sarahrose Webster, Anik Clemens, Jane Winhall-Rice, M. Bento Soares, Cory Yap, Maria de Fatima Bonaldo, Christina H. Smith, Wendy J. Crookes-Goodson, and Carlene K. Chun
- Subjects
lcsh:QH426-470 ,Euprymna scolopes ,lcsh:Biotechnology ,Molecular Sequence Data ,03 medical and health sciences ,Light organ ,lcsh:TP248.13-248.65 ,Genetics ,Juvenile ,Animals ,Aliivibrio fischeri ,Symbiosis ,030304 developmental biology ,Gene Library ,Expressed Sequence Tags ,0303 health sciences ,Expressed sequence tag ,biology ,Base Sequence ,030306 microbiology ,Host (biology) ,cDNA library ,Decapodiformes ,Gene Expression Regulation, Developmental ,Sequence Analysis, DNA ,biology.organism_classification ,Vibrio ,lcsh:Genetics ,Biotechnology ,Research Article - Abstract
Background Biologists are becoming increasingly aware that the interaction of animals, including humans, with their coevolved bacterial partners is essential for health. This growing awareness has been a driving force for the development of models for the study of beneficial animal-bacterial interactions. In the squid-vibrio model, symbiotic Vibrio fischeri induce dramatic developmental changes in the light organ of host Euprymna scolopes over the first hours to days of their partnership. We report here the creation of a juvenile light-organ specific EST database. Results We generated eleven cDNA libraries from the light organ of E. scolopes at developmentally significant time points with and without colonization by V. fischeri. Single pass 3' sequencing efforts generated 42,564 expressed sequence tags (ESTs) of which 35,421 passed our quality criteria and were then clustered via the UIcluster program into 13,962 nonredundant sequences. The cDNA clones representing these nonredundant sequences were sequenced from the 5' end of the vector and 58% of these resulting sequences overlapped significantly with the associated 3' sequence to generate 8,067 contigs with an average sequence length of 1,065 bp. All sequences were annotated with BLASTX (E-value < -03) and Gene Ontology (GO). Conclusion Both the number of ESTs generated from each library and GO categorizations are reflective of the activity state of the light organ during these early stages of symbiosis. Future analyses of the sequences identified in these libraries promise to provide valuable information not only about pathways involved in colonization and early development of the squid light organ, but also about pathways conserved in response to bacterial colonization across the animal kingdom.
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- 2006
28. Risk-Based Quality Management: A Case for Centralized Monitoring.
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Stansbury N, Branco D, McDavid C, Stewart J, Surdam K, Olson N, Perry J, Liska J, Phillips L, Coogan A, Adelfio A, and Garson L
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Since 2019, the Association of Clinical Research Organizations has conducted a landscape survey of risk based quality management (RBQM) adoption in clinical trials. Here, we present data from four years of surveys, with an emphasis on the most recent: the 2022 survey included data from 4958 trials across seven contract research organizations, of which 1004 were new studies started in 2022. Results indicate that while overall risk assessment adoption is strong, it is lagging in other risk-based components which suggests companies are not deriving the full expected benefits of performing a risk assessment and mitigation process to their trials. The 2022 study also suggests new study starts showing promising traction, with adoption hovering near 50% for most RBQM elements. At the same time, the survey suggests industry has mixed views on the potential value of quality tolerance limits (QTLs). Ultimately, centralized monitoring is being underutilized despite the potential of increased patient safety oversight and improved data quality. The authors of this paper developed a case study based on a trial in clinicaltrials.gov to demonstrate how RBQM adoption could include the key RBQM elements such as centralized monitoring, reduced source data review and source data verification as well as implementation of QTLs in a real-world scenario. The authors believe the clinical trial industry has an obligation to utilize centralized monitoring to produce more efficient and effective clinical trials and will make a case to do so in this paper., Competing Interests: Declarations. Conflict of interest: The authors declare no competing interests., (© 2024. The Author(s).)
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- 2024
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29. Competency-Based Medical Education at Scale: A Road Map for Transforming National Systems of Postgraduate Medical Education.
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Karpinski J, Stewart J, Oswald A, Dalseg TR, Atkinson A, and Frank JR
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- Humans, Canada, Competency-Based Education, Curriculum, Education, Medical, Surgeons
- Abstract
In the past decade, the Canadian system of postgraduate medical education has been transformed with the implementation of a new approach to competency based medical education called Competence by Design. The Royal College of Physicians and Surgeons of Canada (Royal College) developed an approach to time-variable competency based medical education and adapted that design for medical, surgical, and diagnostic disciplines. New educational standards and entrustable professional activities consistent with this approach were co-created with 67 specialties and subspecialties, and implementation was scaled up across 17 universities and over 1000 postgraduate training programs. Partner engagement, systematic design of workshops to create discipline specific competency-based standards of education, and agile adaptation were all key ingredients for success. This paper describes the strategies applied by the Royal College, lessons learned regarding transformative change in the complex system of postgraduate medical education, and the current status of the Competence by Design initiative. The approach taken and lessons learned by the Royal College may be useful for other educators who are planning a transformation to CBME or any other major educational reform., Competing Interests: Individual authors received funding from the Royal College either as staff (JK, JS, JRF) or as consultants (AA, TD, AO)., (Copyright: © 2024 The Author(s).)
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- 2024
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30. Correction: Flow Cytometry Method Validation Protocols.
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Selliah N, Nash V, Eck S, Green C, Oldaker T, Stewart J, Vitaliti A, and Litwin V
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- 2024
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31. Primary small intestinal lymphangiosarcoma in a dog presenting with a segmental partial mesenteric volvulus.
- Author
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Ballarini L, Stewart J, Fleming K, and Matchwick A
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- Dogs, Animals, Intestine, Small pathology, Tomography, X-Ray Computed, Intestinal Volvulus veterinary, Intestinal Volvulus diagnosis, Lymphangiosarcoma veterinary, Dog Diseases pathology
- Abstract
A 3-year-old Great Dane presented with a history of chronic vomiting and diarrhoea. Abdominal computed tomography followed by exploratory laparotomy revealed a perforated, segmental partial mesenteric volvulus, affecting an abnormal section of distal jejunum, which was resected. Histopathology and immunohistochemistry results were consistent with jejunal lymphangiosarcoma. This case represents the first report of primary small intestinal lymphangiosarcoma in dogs and the importance of immunohistochemistry for definitive diagnosis., Competing Interests: Declaration of competing interests The authors declared no conflicts of interest in relation to the research, authorship and publication of this article., (Crown Copyright © 2023. Published by Elsevier Ltd. All rights reserved.)
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- 2024
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32. Flow Cytometry Method Validation Protocols.
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Selliah N, Nash V, Eck S, Green C, Oldaker T, Stewart J, Vitaliti A, and Litwin V
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- Flow Cytometry, Academies and Institutes, Biological Assay, Research Design, Clinical Laboratory Services
- Abstract
Analytical method validation provides a means to ensure that data are credible and reproducible. This article will provide a brief introduction to analytical method validation as applied to cellular analysis by flow cytometry, along with practical procedures for four different types of validation. The first, Basic Protocol 1 (the limited validation protocol), is recommended for research and non-regulated laboratories. Next, Basic Protocol 2) presents a reasonable, fit-for-purpose validation approach appropriate for biopharma and research settings. Basic Protocol 3 addresses the type of validation performed in clinical laboratories for moderate-risk tests developed in house. Finally, Basic Protocol 4 describes the process that should be applied whenever a method is being transferred from one facility to another. All four validation plans follow the fit-for-purpose validation approach, in which the validation parameters are selected based on the intended use of the assay. These validation protocols represent the minimal requirement and may not be applicable for every intended use such as high-risk clinical assays or data to be used as a primary endpoint in a clinical trial. The recommendations presented here are consistent with the white papers published by the American Association of Pharmaceutical Scientists and the International Clinical Cytometry Society, as well as with Clinical Laboratory Standards Institute Guideline H62: Validation of Assays Performed by Flow Cytometry (CLSI, 2021). © 2023 Wiley Periodicals LLC. Basic Protocol 1: Limited validation Basic Protocol 2: Fit-for-purpose validation for biopharma and research settings Basic Protocol 3: Validation for moderate clinical risk laboratory developed tests Basic Protocol 4: Transfer validation., (© 2023 Wiley Periodicals LLC.)
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- 2023
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33. Exploring the role of neuronal-enriched extracellular vesicle miR-93 and interoception in major depressive disorder.
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Burrows K, Figueroa-Hall L, Stewart J, Alarbi A, Kuplicki R, Hannafon B, Tan C, Risbrough V, McKinney B, Ramesh R, Victor T, Aupperle R, Savitz J, Teague K, Khalsa S, and Paulus M
- Abstract
Major depressive disorder (MDD) is associated with interoceptive processing dysfunctions, but the molecular mechanisms underlying this dysfunction are poorly understood. This study combined brain Neuronal-Enriched Extracellular Vesicle (NEEV) technology and serum markers of inflammation and metabolism with Functional Magnetic Resonance Imaging (fMRI) to identify the contribution of gene regulatory pathways, in particular micro-RNA (miR) 93, to interoceptive dysfunction in MDD. Individuals with MDD ( n = 44) and healthy comparisons (HC; n = 35) provided blood samples and completed an interoceptive attention task during fMRI. EVs were separated from plasma using a precipitation method. NEEVs were enriched by magnetic streptavidin bead immunocapture utilizing a neural adhesion marker (CD171) biotinylated antibody. NEEV specificities were confirmed by ow cytometry, western blot, particle size analyzer, and transmission electron microscopy. NEEV small RNAs were purified and sequenced. Results showed that: (1) MDD exhibited lower NEEV miR-93 expression than HC; (2) within MDD but not HC, those individuals with the lowest NEEV miR-93 expression had the highest serum concentrations of interleukin (IL)-1 receptor antagonist, IL-6, tumor necrosis factor, and leptin; and (3) within HC but not MDD, those participants with the highest miR-93 expression showed the strongest bilateral dorsal mid-insula activation. Since miR-93 is regulated by stress and affects epigenetic modulation by chromatin reorganization, these results suggest that healthy individuals but not MDD participants show an adaptive epigenetic regulation of insular function during interoceptive processing. Future investigations will need to delineate how specific internal and external environmental conditions contribute to miR-93 expression in MDD and what molecular mechanisms alter brain responsivity to body-relevant signals.
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- 2023
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34. Risk-Based Monitoring in Clinical Trials: 2021 Update.
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Adams A, Adelfio A, Barnes B, Berlien R, Branco D, Coogan A, Garson L, Ramirez N, Stansbury N, Stewart J, Worman G, Butler PJ, and Brown D
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- Humans, Risk Assessment, Surveys and Questionnaires, Research Design
- Abstract
Clinical trial quality depends on ensuring participant safety and data integrity, which require careful management throughout the trial lifecycle, from protocol development to final data analysis and submission. Recent developments-including new regulatory requirements, emerging technologies, and trial decentralization-have increased adoption of risk-based monitoring (RBM) and its parent framework, risk-based quality management (RBQM) in clinical trials. The Association of Clinical Research Organizations (ACRO), recognizing the growing importance of these approaches, initiated an ongoing RBM/RBQM landscape survey project in 2019 to track adoption of the eight functional components of RBQM. Here we present results from the third annual survey, which included data from 4889 clinical trials ongoing in 2021. At least one RBQM component was implemented in 88% of trials in the 2021 survey, compared with 77% in 2020 and 53% in 2019. The most frequently implemented components in 2021 were initial and ongoing risk assessments (80 and 78% of trials, respectively). Only 7% of RBQM trials were Phase IV, while the proportions of Phase I-III trials ranged 27-36%. Small trials (< 300 participants) accounted for 60% of those implementing RBQM. The therapeutic areas with the largest number of RBQM trials were oncology (38%), neurology (10%), and infectious diseases (9%). The 2021 survey confirmed a pattern of increasing RBM/RBQM adoption seen in earlier surveys, with risk assessments, which have broad regulatory support, driving RBQM growth; however, one area requiring further development is implementation of centralized monitoring combined with reductions in source data verification (SDV) and source data review (SDR)., (© 2023. The Author(s).)
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- 2023
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35. Implementation of a comprehensive pharmacy-driven immunization care process model in a pediatric cystic fibrosis clinic.
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Zobell JT, Moss J, Creelman J, Christensen R, Jensen B, Stewart J, Ameel K, and Asfour F
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- Humans, Child, Retrospective Studies, Immunization, Cystic Fibrosis, Papillomavirus Infections, COVID-19, Pharmacy
- Abstract
Introduction: Members of an integrated pharmacy team (pharmacists and pharmacy technicians) have roles that have been identified in the literature as part of the multi-disciplinary cystic fibrosis (CF) care team. One role that has not specifically addressed is the administration of routine and recommended immunizations to people with CF (PwCF). According to care guidelines, PwCF of all ages should be provided all age-appropriate and recommended immunizations. Pharmacists and pharmacy technicians can administer immunizations per state laws. The Primary Children's CF Center decided to implement a comprehensive pharmacy-driven immunization care process model to impact immunization rates., Methods: A 24-month retrospective analysis was conducted with pediatric (≤18 years) PwCF at the Primary Children's CF Center. The primary outcome measures were the percentage (%) of PwCF who received PPSV23, and/or HPV, and/or meningococcal conjugate vaccine (MCV) immunizations 1-year post-care process model implementation (October 1, 2021, to September 30, 2022) as compared to baseline values. The secondary outcome measures are the total number of immunizations, the number of each immunization provided, and the financial impact of pharmacy-driven immunization care process model 1-year post-implementation., Results: During the 1-year post-care process model implementation (October 1, 2021, to September 30, 2022), a total of 523 immunizations were provided to 243 pediatric PwCF. The most frequent immunizations provided were PPSV23 (160/523, 31%) and Coronavirus Disease 2019 (COVID-19) (154/523, 29%). The baseline percentages of eligible PwCF of PPSV23, HPV, and MCV were 27% (58/217), 43% (32/74), and 24% (8/34), respectively. The 1-year post-implementation percentages of PPSV23, HPV, and MCV were 99% (217/218, p < 0.00001), 91% (67/74, p < 0.00001), and 97% (33/34, p < 0.00001), respectively. For COVID-19 immunizations, 56% of eligible PwCF (181/321) have received their first dose. Of these 181 PwCF, 70% (126/181) have received at least one dose of their primary series or booster during the 1-year post-implementation period. The rate of those PwCF who have received at least one dose of a COVID-19 immunization from the age of 6 months to 4 years, 5-11 years, and 12-18 years, was 37% (30/82), 60% (78/129), and 66% (73/110), respectively. For the financial impact generated during the 1-year immunization care process model post-implementation period, 404 non-VFC immunizations were given for an estimated profit of $11,930., Conclusions: The implementation of a pharmacy-driven immunization care process model is a way for integrated pharmacy teams to evolve with the CF center care model and have a role expansion in the care provided to PwCF., (© 2023 Wiley Periodicals LLC.)
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- 2023
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36. Social Support and Parental Conflict as Predictors of Outcomes of Group Cognitive Behavioral Therapy for Adolescent Depression.
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Argiros A, Venanzi L, Dao A, Dickey L, Herman N, Pegg S, Hill K, Stewart J, and Kujawa A
- Abstract
Group cognitive behavioral therapy (CBT) is an effective treatment for adolescent depression, but outcomes vary. Our goal was to examine interpersonal factors that predict response to group CBT for adolescent depression using a broad range of outcomes, including depressive symptoms, session attendance, treatment completion, engagement, and improvement. Seventy adolescents (age 14-18) with depression completed self-report measures of social support and parental conflict and were offered an established 16-session group CBT program. Correlation and regression analyses were conducted for interpersonal predictors and CBT outcomes. Accounting for pre-treatment depressive symptoms, fewer social supports predicted lower likelihood of finishing treatment and less clinician-rated improvement. Greater pre-treatment parental conflict predicted fewer sessions attended, lower clinician-rated engagement, and less clinician-rated improvement. Results highlight the need to consider interpersonal difficulties in CBT, as they may present a barrier to treatment attendance, engagement, and improvement., Competing Interests: Conflicts of InterestThe authors declare no conflicts of interest., (© Springer Nature Switzerland AG 2023, Springer Nature or its licensor (e.g. a society or other partner) holds exclusive rights to this article under a publishing agreement with the author(s) or other rightsholder(s); author self-archiving of the accepted manuscript version of this article is solely governed by the terms of such publishing agreement and applicable law.)
- Published
- 2023
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37. A positive parenting program to enhance positive affect in children of previously depressed mothers.
- Author
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Cullum KA, Goodman SH, Garber J, Korelitz K, Sutherland S, and Stewart J
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- Attention, Child, Female, Humans, Infectious Disease Transmission, Vertical, Mothers psychology, Parenting psychology
- Abstract
Children of mothers with a history of depression are at heightened risk for developing depression and other maladaptive outcomes. Deficits in parenting are one putative mechanism underlying this transmission of risk from mother to child. The present study evaluated whether a brief intervention with mothers with a history of depression produced greater use of positive parenting behaviors and an increase in observed positive affect in their 8- to 10-year-old children. Mothers with a history of depression ( n = 65) were randomly assigned to either a positive parenting intervention or an attention control intervention condition. In addition, a comparison group of 66 mothers with no history of depression was evaluated one time. Results revealed significant increases in positive parenting behaviors (e.g., active listening, praise) immediately postintervention in mothers randomized to the positive parenting intervention as compared to those in the attention control condition. Children of mothers in the positive parenting intervention showed increases in positive affect as compared to children of mothers in the attention control intervention. Increases in mothers' active listening and smiling/laughing significantly predicted increases in children's positive affect. The intervention did not increase the rate of children's moment-by-moment positive affect contingent on mothers' positive parenting behaviors. This study showed the short-term effectiveness of a brief parenting intervention for enhancing interactions between mothers with a history of depression and their children by directly targeting mothers' positive parenting and, indirectly, children's expressions of positive affect. (PsycInfo Database Record (c) 2022 APA, all rights reserved).
- Published
- 2022
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38. Best Practices in Dementia Care: A Review of the Grey Literature on Guidelines for Staffing and Physical Environment in Long-Term Care.
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Seetharaman K, Chaudhury H, Kary M, Stewart J, Lindsay B, and Hudson M
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- Canada, Gray Literature, Humans, Workforce, Dementia therapy, Long-Term Care
- Abstract
In its first national strategy on dementia, the Government of Canada has highlighted the need to improve quality of care for individuals living with dementia, with emphasis on following best practices and evidence in care delivery and providing care staff access to education and training. It is also known that the design of the physical environment of care homes is integral to the care experience of individuals living with dementia. Therefore, this study aims to identify the best national and international practices implemented in care homes for people living with dementia in: (1) education, training, staffing, and care practices; and (2) environmental design and physical infrastructure, through the review of relevant grey literature. This article highlights key recommendations for improving the quality of care for residents living with dementia in care homes, such as: (1) facilitating translation of training into practice, (2) maintaining consistent staffing levels, and (3) designing care homes to facilitate wayfinding, accessibility, safety, comfort, appropriate sensory stimulation, familiarity, and homelikeness. The findings from this review are expected to inform the development of guidelines for a provincial dementia-friendly care home designation program and various advocacy efforts to help achieve the objectives of the national strategy on dementia.
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- 2022
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39. Quality improvement and rapid PDSA cycles to maintain routine surveillance of pulmonary pathogens during the COVID-19 pandemic in a pediatric cystic fibrosis clinic.
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Hamilton JL, Snuggerud AE, Meihls SM, Toledo H, and Stewart J
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- Child, Humans, Pandemics, Quality Improvement, COVID-19, Cystic Fibrosis, Telemedicine
- Abstract
Background: The COVID-19 pandemic necessitated immediate transition from in person to telehealth encounters; novel nursing practices were needed to ensure that children with cystic fibrosis (CF) receive care that approximates evidence-based guidelines., Local Problem: The aim was to ensure that as many children as possible received routine surveillance of pulmonary pathogens by a CF culture sputum culture during a pandemic., Methods: Multiple Plan-Do-Study-Act (PDSA) cycles were utilized to implement practice change over four months., Interventions: Cultures were obtained via curbside appointment with a registered nurse (RN) or at the patients' home with mailed equipment., Results: 133 cultures obtained: 50.37% (67) by RN collection curbside and 49.62% (66) by self/caregiver at home. 120 culture swabs or sterile cups were mailed; 55% (66) were returned. Cost of mailing equipment was $760.16., Conclusion: Nursing utilization of PDSA cycles developed novel processes that ensured guideline-based care during the initial months of the pandemic., (Copyright © 2021 Elsevier Inc. All rights reserved.)
- Published
- 2022
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40. Prevalence and implications of frailty in acute stroke: systematic review & meta-analysis.
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Burton JK, Stewart J, Blair M, Oxley S, Wass A, Taylor-Rowan M, and Quinn TJ
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- Aged, Frail Elderly, Humans, Prevalence, Risk Factors, Frailty diagnosis, Frailty epidemiology, Stroke diagnosis, Stroke epidemiology, Stroke therapy
- Abstract
Background: frailty is common in older adults and associated with poor outcomes following illness. Although stroke is predominantly a disease of older people, our knowledge of frailty in stroke is limited. We aimed to collate the literature on acute stroke and frailty to estimate the prevalence of pre-stroke frailty and its associations with outcomes., Methods: paired researchers searched multidisciplinary electronic databases for papers describing frailty and acute stroke. We assessed risk of bias using Newcastle-Ottawa tools appropriate to study design. We created summary estimates of pre-stroke frailty using random effects models. We collated whether studies reported significant positive associations between frailty and clinical outcomes in adjusted models., Results: we included 14 studies (n = 27,210 participants). Seven studies (n = 8,840) used a frailty index approach, four studies (n = 14,924) used Hospital Frailty Risk Scores. Pooled prevalence of pre-stroke frailty was 24.6% (95% confidence interval, CI: 16.2-33.1%; low quality evidence, downgraded due to heterogeneity, bias). Combining frailty and pre-frailty (nine studies, n = 23,827), prevalence of any frailty syndrome was 66.8% (95%CI: 49.9-83.7%). Seven studies were at risk of bias, from participant selection or method of frailty assessment. Pre-stroke frailty was associated with all adverse outcomes assessed, including longer-term mortality (positive association in 6 of 6 studies reporting this outcome; odds ratio: 3.75 [95%CI: 2.41-5.70]), length of admission (3 of 4 studies) and disability (4 of 6 studies)., Conclusions: despite substantial heterogeneity, whichever way it is measured, frailty is common in patients presenting with acute stroke and associated with poor outcomes. This has implications for the design of stroke services and pathways., (© The Author(s) 2022. Published by Oxford University Press on behalf of the British Geriatrics Society. All rights reserved. For permissions, please email: journals.permissions@oup.com.)
- Published
- 2022
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41. Liposomal bupivacaine addition versus standard bupivacaine alone for colorectal surgery: a randomized controlled trial.
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Yeap YL, Wolfe J, Stewart J, McCutchan A, Chawla G, Robb B, Holcomb B, and Vickery B
- Subjects
- Analgesics, Opioid, Anesthetics, Local, Humans, Pain, Postoperative drug therapy, Pain, Postoperative prevention & control, Prospective Studies, Bupivacaine, Colorectal Surgery
- Abstract
Aim: This study evaluated use of liposomal bupivacaine (LB) versus standard bupivacaine (SB) alone in quadratus lumborum (QL) blocks for laparoscopic colorectal surgery. Materials & methods: In this prospective, randomized controlled trial, patients received QL1 blocks with either LB (40 ml 0.125% SB plus 20 ml of LB) or SB (60 ml of 0.25% SB) with 30 ml per side. Opioid usage, pain scores, side effects and other medications were recorded. Results: For 78 patients (38 LB; 40 SB), all parameters were similar between groups, except that the LB group had a higher 48 h need for metoclopramide. Conclusion: LB provided no analgesic benefit over SB alone for QL blocks. Clinical Trials registration number: NCT03702621.
- Published
- 2022
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42. Advanced practice providers in pediatric pulmonary.
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Hamilton JL and Stewart J
- Subjects
- Child, Humans, Clinical Competence
- Published
- 2021
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43. Gross post-mortem and histological features in 27 horses with confirmed lumbosacral region pain and five control horses: A descriptive cadaveric study.
- Author
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Quiney L, Stewart J, Routh J, and Dyson S
- Abstract
Background: There is a lack of understanding of the pathological and/or physiological nature of lumbosacral region pain., Objectives: To describe the gross variations of the osseous and soft tissues of the lumbosacral region and report the histological findings of sections of nerve tissue in affected and control horses., Study Design: Descriptive post-mortem case series., Methods: All horses had undergone full clinical and gait assessment, including ridden exercise. Horses with a substantial response to infiltration of local anaesthetic solution around the sacroiliac joint regions were included in the affected group (n = 27). Horses for which the source(s) of pain was confirmed by diagnostic anaesthesia to be distant to the lumbosacral region were included in the control group (n = 5). The pelvic regions were isolated and the soft tissues were assessed grossly. Sections of the lumbosacral plexus and cranial gluteal, sciatic and obturator nerves were examined histologically. The osseous specimens were evaluated for anatomical variants and abnormalities. Data were analysed using descriptive statistics., Results: Gross discolouration of the sciatic or obturator nerves was observed in 7 (26%) affected and no control horses. Grade 3/3 histological abnormality scores were assigned in 22% of nerve sections from affected horses compared with 3% from control horses. Several osseous variants (bifid sacral spinous processes, straight-shaped sacroiliac joint surface, short arrow-shaped sacral alae, left-right asymmetry of sacral alae, sacral curvature, absence of the fourth to fifth and ankylosis of the fifth to sixth lumbar articular process joints, left-right asymmetry of caudocranial position of the fourth to fifth and lumbar-sacral articular process joints) and abnormalities (sacroiliac enthesopathy, extra ventral sacroiliac joint surface, lumbosacral symphyseal periarticular modelling, lumbosacral intertransverse joint pitting lesions) were more frequently observed in affected horses., Main Limitations: Both control and affected horses may have had preclinical abnormalities., Conclusions: Lumbosacral region pain may reflect the presence of a number of pathological changes. Neural pain may play an important role in some horses., (© 2021 EVJ Ltd.)
- Published
- 2021
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44. Global Health Action at 15 - revisiting its rationale.
- Author
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Wall S, Emmelin M, Krantz I, Nilsson M, Norström F, Schröders J, Stewart Williams J, and Östergren PO
- Subjects
- Humans, Global Health
- Published
- 2021
- Full Text
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45. Factors contributing to the uptake of childhood vaccination in Galkayo District, Puntland, Somalia.
- Author
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Abdullahi MF, Stewart Williams J, Sahlèn KG, Bile K, and Kinsman J
- Subjects
- Adult, Child, Fathers, Female, Focus Groups, Humans, Male, Qualitative Research, Somalia, Child Health ethnology, Decision Making, Health Knowledge, Attitudes, Practice ethnology, Health Personnel psychology, Parents psychology, Vaccination
- Abstract
Background: As in many Sub-Saharan African countries, the health system in Somalia is not operating at the capacity needed to lift childhood vaccination coverage to ninety percent or above, as recommended by United Nations Children's Fund. Current national estimates of coverage for the six major vaccine preventable childhood diseases range from thirty to sixty percent. Infectious disease outbreaks continue to pose significant challenges for the country's health authorities., Objective: This important qualitative study, conducted in Galkayo District, Somalia, investigates limiting factors associated with childhood vaccination uptake from the perspective of both communities and health care workers., Methods: Qualitative information was collected through six focus group discussions with parents (n = 48) and five one-to-one interviews with health workers (n = 15) between March and May 2017, in three settings in the Galkayo District - Galkayo city, Bayra and Bacadwayn., Results: From a health system perspective, the factors are: awareness raising, hard to reach areas, negative attitudes and perceived knowledge of health workers, inadequate supplies and infrastructure, and missed vaccination opportunities. From the perspective of individuals and communities the factors are: low trust in vaccines, misinterpretation of religious beliefs, vaccine refusals, Somalia's patriarchal system and rumours and misinformation. Parents mostly received immunization information from social mobilizers and health facilities. Fathers, who are typically family decision-makers, were poorly informed. The findings highlight the need for in-service training to enable health workers to improve communication with parents, particularly fathers, peripheral communities and local religious leaders., Conclusions: Enhancing knowledge and awareness of vaccination among parents is crucial. Fathers' involvement is lacking. This may be boosted by highlighting fathers' obligation to protect their children's health through vaccination. It is also important that men engage with the wider community in decision-making and advance towards the global vaccination targets.
- Published
- 2020
- Full Text
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46. Effect of Prior Formal Education on Successful Thoracic Epidural Placement By Anesthesia Residents.
- Author
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Yeap YL, Randolph T, Lemmon AJ, Mann MD, Stewart J, and Wolfe JW
- Subjects
- Clinical Competence, Humans, Prospective Studies, Anesthesia, Epidural, Anesthesiology education, Internship and Residency
- Abstract
Objective: Catheter placement for thoracic epidural analgesia (TEA) is technically challenging; however, methods for teaching this technique to anesthesia residents have not been well-studied. The present study aimed to determine optimal teaching methods for proficient TEA catheter placement by comparing video-based formal resident education with traditional bedside training by attending physicians., Design: Prospective, randomized study., Setting: Large academic hospital, single institution., Participants: The study comprised 76 postgraduate year 3 and 4 anesthesiology residents (38 intervention, 38 control)., Interventions: Formal education included an instructional video on proper TEA technique., Measurements and Main Results: Measures of proficiency in TEA catheter placement included the time needed to complete the procedure successfully and the success of placement as indicated by patient confirmation. Residents who received formal video instruction had similar success in catheter placement and similar procedure times compared with the traditionally trained residents. The overall success rate was 99.2%, with faculty intervention required in only 17% of cases. More experienced residents (ie, having placed more epidural catheters) were faster at TEA catheter placement., Conclusions: Formal video education for TEA catheter placement provided no additional improvement of resident proficiency compared with traditional training at a high-volume academic center. The success rate was very high in this group of residents; however, experiences at other institutions may vary. Future studies are needed to determine optimum teaching strategies for TEA., Competing Interests: Conflict of Interest The authors declare no conflicts of interest related to this research., (Copyright © 2020 Elsevier Inc. All rights reserved.)
- Published
- 2020
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47. Physical activity during COVID-19 quarantine.
- Author
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Shahidi SH, Stewart Williams J, and Hassani F
- Subjects
- Child, Humans, COVID-19, Exercise psychology, Quarantine
- Published
- 2020
- Full Text
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48. Substance Use Disorders and COVID-19: Multi-Faceted Problems Which Require Multi-Pronged Solutions.
- Author
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Jemberie WB, Stewart Williams J, Eriksson M, Grönlund AS, Ng N, Blom Nilsson M, Padyab M, Priest KC, Sandlund M, Snellman F, McCarty D, and Lundgren LM
- Abstract
COVID-19 shocked health and economic systems leaving millions of people without employment and safety nets. The pandemic disproportionately affects people with substance use disorders (SUDs) due to the collision between SUDs and COVID-19. Comorbidities and risk environments for SUDs are likely risk factors for COVID-19. The pandemic, in turn, diminishes resources that people with SUD need for their recovery and well-being. This article presents an interdisciplinary and international perspective on how COVID-19 and the related systemic shock impact on individuals with SUDs directly and indirectly. We highlight a need to understand SUDs as biopsychosocial disorders and use evidence-based policies to destigmatize SUDs. We recommend a suite of multi-sectorial actions and strategies to strengthen, modernize and complement addiction care systems which will become resilient and responsive to future systemic shocks similar to the COVID-19 pandemic., (Copyright © 2020 Jemberie, Stewart Williams, Eriksson, Grönlund, Ng, Blom Nilsson, Padyab, Priest, Sandlund, Snellman, McCarty and Lundgren.)
- Published
- 2020
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49. Health systems responsiveness among older adults: Findings from the World Health Organization Study on global AGEing and adult health.
- Author
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Stewart Williams J, Myléus A, Chatterji S, and Valentine N
- Subjects
- Aged, China, Cross-Sectional Studies, Ghana, Humans, Independent Living, India, Middle Aged, Russia, South Africa, World Health Organization, Delivery of Health Care organization & administration, Global Health
- Abstract
Health system responsiveness is an indicator that can be used for evaluating how well healthcare systems respond to people's needs in non-clinical areas such as communication, autonomy and confidentiality. This study analyses health system responsiveness from the perspective of community-dwelling adults aged 50 and over in China, Ghana, India, the Russian Federation and South Africa using cross-sectional data from the World Health Organization Study on global AGEing and adult health. The aim is to assess and compare how individual, health condition and healthcare factors impact differently on outpatient and inpatient responsiveness. Poor responsiveness is measured according to participants' responses to questions on a five-point Likert scale. Five univariate and multiple logistic regression models test associations between individual, health condition and healthcare factors and poor responsiveness. The final model adjusts for country. Key results are that travel time is a major contributor to poor responsiveness across all countries. Similarly there are wealth inequalities in responsiveness. However no clear difference in responsiveness was observed in presentations for chronic versus other types of conditions. This study provides an interesting baseline on older patients' perceived treatment within outpatient and inpatient facilities in five diverse low- and middle-income countries.
- Published
- 2020
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50. Randomized Prospective Study Evaluating Single-Injection Paravertebral Block, Paravertebral Catheter, and Thoracic Epidural Catheter for Postoperative Regional Analgesia After Video-Assisted Thoracoscopic Surgery.
- Author
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Yeap YL, Wolfe JW, Backfish-White KM, Young JV, Stewart J, Ceppa DP, Moser EAS, and Birdas TJ
- Subjects
- Adult, Catheters, Humans, Pain Measurement, Pain, Postoperative diagnosis, Pain, Postoperative drug therapy, Pain, Postoperative prevention & control, Prospective Studies, Thoracic Surgery, Video-Assisted, Analgesia, Anesthesia, Epidural, Nerve Block
- Abstract
Objective: Video-assisted thoracoscopic surgery (VATS) has improved patient outcomes; however, postoperative pain remains potentially severe. The objective of this study was to compare adjunct analgesic modalities for VATS, including paravertebral nerve blockade (PVB) and thoracic epidural anesthesia (TEA)., Design: Prospective, randomized trial., Setting: Large academic hospital, single institution., Participants: Adult patients undergoing VATS., Interventions: Ultrasound-guided PVB catheter, ultrasound-guided single-injection PVB, or TEA., Measurements and Main Results: Postoperative visual analog scale pain scores (at rest and with knee flexion) and opioid usage were recorded. Pain scores (with movement) for the TEA group were lower than those for either PVB group at 24 hours (p ≤ 0.008) and for the PVB catheter group at 48 hours (p = 0.002). Opioid use in TEA group was lower than that for either PVB group at 24 and 48 hours (p < 0.001) and 72 hours (p < 0.05). Single-injection PVB was faster compared with PVB catheter placement (6 min v 12 min; p < 0.001) but similar to TEA (5 min). Patient satisfaction, nausea, sedation, and 6-month postsurgical pain did not differ between groups., Conclusions: TEA led to lower pain scores and opioid requirement for VATS procedures compared with PVB techniques. Single-injection PVB was faster and equally as effective as PVB catheter, and it led to similar patient satisfaction as TEA; therefore, it should be considered in patients who are not ideal candidates for TEA., (Copyright © 2020 The Authors. Published by Elsevier Inc. All rights reserved.)
- Published
- 2020
- Full Text
- View/download PDF
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