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3. Art

Author

Sonya Clark

Subjects
Marketing, Pharmacology, Organizational Behavior and Human Resource Management, Strategy and Management, Drug Discovery, Pharmaceutical Science
Published
2021
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4. Evaluation of a dementia awareness game for health professions students in Northern Ireland: a pre-/post-test study

Author

Stephanie Craig, Heather E. Barry, Gillian Carter, Patrick Stark, Gary Mitchell, Sonya Clarke, and Christine Brown Wilson

Subjects
Dementia, Nurse education, Pharmacy education, Serious games, Students, Pedagogical research, Special aspects of education, LC8-6691, Medicine
Abstract

Abstract Background Dementia is a prevalent global health issue, necessitating comprehensive education for healthcare practitioners and students. Nursing and pharmacy students, provide support across healthcare settings often working as frontline caregivers. Therefore, it is imperative to equip these students with a profound understanding of dementia. The aim of this study was to evaluate whether a serious dementia game co-designed with stakeholders, students, and people living with dementia improved the attitudes of nursing and pharmacy students. Methods A pretest-posttest design was used to assess the attitudes of health professions students (nursing and pharmacy) towards dementia. The Approaches to Dementia Questionnaire (ADQ) was administered before and after playing a serious Dementia Game. The ADQ measured the total score, Hope subscale, and Recognition of Personhood subscale. Matched pairs t-test was used for analysis conducted with IBM SPSS statistics 27. Results A diverse cohort of 505 participants from one university in Northern Ireland participated, with 461 matched pairs used for analysis. Both nursing and pharmacy students demonstrated a significant increase in overall dementia attitudes post-gameplay, with nursing students showing an increase from 79.69 to 83.59 and pharmacy students from 75.55 to 79.86. Subscales for Hope (Nursing = 28.77 to 31.22, Pharmacy = 26.65 to 29.20). and Recognition of Personhood also exhibited significant improvement (Nursing = 50.93 to 52.38, Pharmacy = 48.89 to 50.67). Demographic data revealed predominantly female participants, a lack of personal connections to dementia, and varied training experiences. Discussion The study highlights the efficacy of the serious Dementia Game in enhancing attitudes to dementia amongst health professions students, indicating its potential as an educational tool. The study contributes to the growing body of evidence supporting serious games and gamification in healthcare education.

Published
2024
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5. 3′ Branch ligation: a novel method to ligate non-complementary DNA to recessed or internal 3′OH ends in DNA or RNA

Author

Xia Zhao, Xu Xu, Hanjie Shen, Xun Xu, Han Ren, Helena Perazich, Lin Wang, Xiaojue Wang, Yang Xi, Wenwei Zhang, Sonya Clark, Alayna Albert, Ou Wang, Ao Chen, Brock A. Peters, Killeen Kirkconnell, Radoje Drmanac, Evan Hurowitz, Andrei Alexeev, Wang Weimao, and Yuan Jiang

Subjects
0106 biological sciences, DNA Ligases, Computational biology, novel ligation, Biology, 01 natural sciences, DNA sequencing, 03 medical and health sciences, chemistry.chemical_compound, Adapter (genetics), Complementary DNA, Genetics, Humans, A-DNA, Molecular Biology, 030304 developmental biology, chemistry.chemical_classification, 0303 health sciences, DNA ligase, RNA, High-Throughput Nucleotide Sequencing, General Medicine, DNA, Full Papers, Cell biology, genomic DNA, T4 DNA ligase, chemistry, NGS, molecular tool, Nucleic acid, Ligation, Genetic Engineering, 010606 plant biology & botany
Abstract

Nucleic acid ligases are crucial enzymes that repair breaks in DNA or RNA during synthesis, repair and recombination. Various molecular tools have been developed using the diverse activities of DNA/RNA ligases. Herein, we demonstrate a non-conventional ability of T4 DNA ligase to join 5’ phosphorylated blunt-end double-stranded DNA to DNA breaks at 3’ recessive ends, gaps, or nicks to form a 3’ branch structure. Therefore, this base pairing-independent ligation is termed 3’ branch ligation (3’BL). In an extensive study of optimal ligation conditions, similar to blunt-end ligation, the presence of 10% PEG-8000 in the ligation buffer significantly increased ligation efficiency. A low level of nucleotide preference was observed at the junction sites using different synthetic DNAs. Furthermore, we discovered that T4 DNA ligase efficiently ligated DNA to the 3’ recessed end of RNA, not to that of DNA, in a DNA/RNA hybrid, whereas RNA ligases are less efficient in this reaction. These novel properties of T4 DNA ligase can be utilized as a broad molecular technique in many important applications. We performed a proof-of-concept study of a new directional tagmentation protocol for next generation sequencing (NGS) library construction that eliminates inverted adapters and allows sample barcode insertion adjacent to genomic DNA. 3’BL after single transposon tagmentation can theoretically achieve 100% usable template, and our empirical data demonstrate that the new approach produced higher yield compared with traditional double transposon or Y transposon tagmentation. We further explore the potential use of 3’BL for preparing targeted RNA NGS libraries with mitigated structure-based bias and adapter dimer problems.

Published
2018

6. Large Scale Synthetic Site Saturation GPCR Libraries Reveal Novel Mutations That Alter Glucose Signaling

Author

William M. Shaw, Ross Kettleborough, Tom Ellis, David Öling, Niklas Larsson, Mark Wigglesworth, Sonya Clark, and Lina Lawenius

Subjects
0301 basic medicine, Saccharomyces cerevisiae Proteins, Computer science, Biomedical Engineering, Down-Regulation, Saccharomyces cerevisiae, Computational biology, Oligonucleotide synthesis, Biochemistry, Genetics and Molecular Biology (miscellaneous), Receptors, G-Protein-Coupled, 03 medical and health sciences, Synthetic biology, 0302 clinical medicine, Saturated mutagenesis, Massively parallel, Gene Library, G protein-coupled receptor, beta-Fructofuranosidase, General Medicine, Protein engineering, Directed evolution, Up-Regulation, Glucose, 030104 developmental biology, Mutagenesis, Site-Directed, Biosensor, 030217 neurology & neurosurgery, Signal Transduction
Abstract

Site saturation mutagenesis (SSM) is a powerful mutagenesis strategy for protein engineering and directed evolution experiments. However, limiting factors using this method are either biased representation of variants, or limiting library size. To overcome these hurdles, we generated large scale targeted synthetic SSM libraries using massively parallel oligonucleotide synthesis and benchmarked this against an error-prone (epPCR) library. The yeast glucose activated GPCR-Gpr1 was chosen as a prototype to evolve novel glucose sensors. We demonstrate superior variant representation and several unique hits in the synthetic library compared to the PCR generated library. Application of this mutational approach further builds the possibilities of synthetic biology in tuning of a response to known ligands and in generating biosensors for novel ligands.

Published
2018
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7. PW 0653 Build your balance: the devleopment of a falls prevention exercise video for older adults in western australia

Author

Catrina Wold, Chloe Macri, Rachel Meade, and Sonya Clark

Subjects
Formative assessment, Gerontology, Resource (project management), Home environment, Relevance (information retrieval), Voice over, Psychology, Quarter (United States coin), Focus group, Balance (ability)
Abstract

Balance is a key risk factor for falls, with one quarter of all falls among older adults caused by poor balance. To increase awareness of the importance of balance in older adults living independently in Western Australia (WA), the Stay On Your Feet®program, provided by Injury Matters and funded by the WA Department of Health, implements the ‘Build Your Balance’ campaign. Following a formative rapid review, a digital video was identified as a relevant resource to educate community members on safe balance exercises. The objective was to develop the Stay On Your Feet® Build Your Balance Exercise Video that meets the needs of the target demographic. Consultations with health professionals (n=15) were undertaken via survey and telephone interviews to identify the needs of a balance resource. To develop the concept, consultation with a physiotherapist and an exercise physiologist was completed, and tested in two community focus groups (n=35) to assess content relevance. Consultations with health professionals concluded the video should contain both a health professional and older adult and needed to address exercise safety and correct technique. Results From the community focus groups reported the preferred format and timeframe of a balance resource as a 15 min DVD. The content was identified as needing to cover the importance of balance exercises that range in fitness levels, safety information, and include a voice over of step-by-step exercise technique. The messaging needed to be positive and the balance exercises should be able to be incorporated into daily routines and exercise programs. The preferred video setting was a home environment and the cast was favoured to be relatable and use ‘everyday people’. To best meet the needs of older adults, the Build Your Balance Exercise Video needs to be relatable, easily incorporated into daily routines and appropriate to complete independently.

Published
2018
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8. Evaluation of a dementia awareness game for undergraduate nursing students in Northern Ireland: a Pre-/Post-Test study

Author

Stephanie Craig, Patrick Stark, Christine Brown Wilson, Gillian Carter, Sonya Clarke, and Gary Mitchell

Subjects
Dementia, Nurse education, Nursing student, Serious game, Pedagogical research, Constructivist pedagogy, Nursing, RT1-120
Abstract

Abstract Introduction Although it is possible to live well with dementia and many individuals with dementia lead active lives with the help of family, friends, and communities, the general impression of dementia is frequently negative. Dementia is a global health issue. Despite this, little research has been done on the effects of innovative dementia education strategies among undergraduate nursing students. The aim of this study was therefore to assess if this serious digital game, originally intended for the public, could increase knowledge about dementia in first-year nursing students. Methods The intervention was a digital serious game called “The Dementia Game”, which was available to students throughout February 2021, to a convenience sample of first-year undergraduate nursing students (n = 560) completing a BSc Honours Nursing Degree programme in one university in Northern Ireland. The game was evaluated using a pretest-posttest design. The questionnaire comprised of a 30- item true- false Alzheimer’s Disease Knowledge Scale (ADKS), which covers risk factors, assessment and diagnosis, symptoms, course, life impact, caregiving and treatment and management. Data were analysed using paired t-tests and descriptive statistics. Results Overall dementia knowledge increased significantly after playing the game. Pre-test to post-test increases were observed across a range of seven categories of dementia knowledge (life impact, risk factors, symptoms, treatment, assessment, caregiving and trajectory), with particularly large increases in knowledge of trajectory and risk factors, as shown using paired t-tests. All pre-test to post-test comparisons were significant at the p

Published
2023
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9. Hair Craft Project

Author

Sonya Clark

Subjects
African american, Craft, Visual Arts and Performing Arts, business.industry, Anthropology, Media studies, Commonwealth, Sociology, business, The arts, Bridge (music), Visual arts, Fine art
Abstract

The Hair Craft Project is an exercise in socially engaged art practices, based on the premise that hairdressing is the primordial textile art form. Sonya Clark, artist and project director, sought to bridge multiple communities: African American hair salons; students, faculty, and alumni of Virginia Commonwealth University’s nationally ranked arts programs; and the broader art community. The project frames the complex craftsmanship, engineering, and skill of hairstylists specializing in African hair-braiding techniques in cultural, historical, and contemporary-art contexts. The Hair Craft Project is innovative in its methods of attracting, building, and bridging audiences while supporting and acknowledging the parallel skills of artists and hairstylists. Through this art form, the project connected cultures, class, and race in Richmond, Virginia, a city with a history of being racially, culturally, and economically divided. The project went on to gain national attention. It was selected by artists Leonardo Drew, Katherina Grosse, and the late founder of Art 21, Susan Sollins, for the Juror’s Grand Prize at Art Prize in Grand Rapids, Michigan, in fall 2014. The Hair Craft Project will travel to the Museum of Fine Arts, Boston, in Fall 2015.

Published
2015
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10. Considering the future direction of children's and young people's nursing

Author

Jim Richardson, Sonya Clark, and Alan Glasper

Subjects
Mental Health Services, Adolescent, Child Welfare, Nurses, Queen (playing card), 03 medical and health sciences, 0302 clinical medicine, Nursing, Humans, Medicine, Nurse education, Child, Education, Nursing, Simulation Training, General Nursing, 030504 nursing, business.industry, Mentoring, Congresses as Topic, United Kingdom, Pediatric Nursing, Mental Health, 030220 oncology & carcinogenesis, George (robot), Academic community, Curriculum, 0305 other medical science, business, Needs Assessment, Forecasting
Abstract

Sonya Clark, Senior Lecturer in Children's Nursing at Queen's University Belfast, Emeritus Professor Alan Glasper, from the University of Southampton and Jim Richardson, Senior Lecturer in Children's Nursing at Kingston University and St George's, University of London, discuss the concerns of the academic community

Published
2017
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11. The National Lung Screening Trial: Overview and Study Design

Author

Natalie Cunningham, Michael Khalili, John Waltz, Ralph Weiben, Deb Gurtner, Linda DeAlmeida, Sanjay Gupta, Sharon Maxfield, Crissy Kibic, Kathleen DeWitt, David DeMets, Walter Allen Bowman, Robert Epstein, Mia Burkhard, Stephen J. Swensen, Hattie Cromwell, Kianoush Rezai, Steadman Sankey, Lisa Scott Wasson, Rita Musanti, Tamim Malbari, Joy Ferola, Qimei He, Patty Trapnell, Melba Francis, Sam Quattlebaum, Joanice Thompson, Ana Birofka, Robin Griggs, Elizabeth Johnson, Margaret R. Spitz, Nicole Richardson, Yuting Liang, Lawrence G. Hutchins, Mirjana Tecmire, Lila Camara, James J. Navin, Eileen Frost, Diane Romano, Carrie Petkus, Eric J. Berns, Pei Jan P Lin, Steve D. Uttecht, Marian Acerra, Lawrence R. Ragard, Leo P. Lawler, Christopher M. Rogers, Alan Lee Goodwin, L. Ellen Martinusen, Melissa Ford, Michael T. Fisher, Beverly Powell, Cindy Lin, Jamie Downs, Brent Fodera, Bonita Wohlers, Michael Brangan, Peggy Bradley, Todd B. Burt, Susan Allen, Shiva Borgheian, Mingying Zeng, Thomas Riley, Danielle Gherardini, Steven Shiff, Olivia Campa, Wahied Gendi, Fang F. Xu, Ivana K. Kazda, Anne Chung, Briar Doi, Helen Price, Maria Vlachou, Alan Morgan, Simone Vuong, Pierre P. Massion, Darcy Watson, Debbie William, Esther Nakano, Karen Broski, David Creed, Melanie Bvorak, Lakisha Hawkins, Gladys Hino, Raymond Dauphinais, Michele Sallas, Helene Shiratori, Venus Brown, Denise Brooks, Heather Porter, Ilana F. Gareen, Tracy Lee, Melissa Cates, Kyle Turner, Tiffanie Hammond, Margaret Paquette, Lorraine Kerchum, Barbara Lewis, Douglas J. Reding, Thomas E. Hartman, Cathy Longden, Melissa Laron, Reza Abaya, Beborah Robertson, J W Semenkovich, Christine Holland, Hugh McGinley, Chani Montalbo, Karen Zubena, Vanessa Ralda, Adam C. Stein, Jennifer Ott, Lawrence M. Kotner, Jing Lee, Arnold Ssali, Michael Young, Quinn A. DeMordaunt, Linda V. White, Steve Dubinett, Pearl Chan, Roxana Phillips, Mallory Kolich, Brent B. Nelson, Phi Do, Jill Spivak, Angele LaFleur, Kesha Smith, Elayne Weslowsky, Patricia Nieters, Maurice LeBlanc, Satinder Singh, Lonna Matthews, Quentin McMullen, Karen Lappe, Sharon Longacre, Cindy Cobb, Jane A. Zehner, Michael Teepe, Pamela M. Marcus, Kathleen Bow, Wendy Francis, Mary Gemmel, Robert S. Fontana, Linda Jurjans, Barbara Ginther, Jonathan B. Clapp, Monica Richel, Scott F. Pickering, Brenda Edwards, Kendrick Looney, Randy Marshall, Roni Atkins, Danielle Wicks, Julie Peterson, Dcanna Cape, Albert J. Cook, Jerry Brekke, Louisa Turner, Larry Stoller, Mark B. Salerno, Bavid E. Midthun, Mark Delano, Minnetta Belyea, Deborah Greene, Jonathan Goldin, Terry Lewis, Virginia Fischer, Andrea Chapman, Shari Jordan, Deb Warren, Demetria Johnson, Rekha Khatri, Lisa Sirianni, Guillermo Geisse, Michael A. Fuchs, Kanya Kumbalasiri, Jeremy J. Erasmus, Vicki Shambaugh, Denise Boyles, Sarah Hallsky, Anna Nanovski, Jill Heinz, Mollie King, Kay Vydareny, Olga Soukhanova, Patricia Rueweler, Perry G. Pernicano, Regina Rendas-Baum, Phyllis Pirotte, Russell Harris, Neil Argyle, Miyoung Kim, June Krebsbach, Audrey Gallego, Sheila Wein, Mukesh F. Karwat, Karla Myra-Bloom, Pamela Byrnes, Mitchell D. Schnall, Hector Ahumada, Eric Sanchez, Donna DesMarais, Julie Maderitz, Cindy Lavergne, Lori Kirchoff, Patricia C. Sanders, Elizabeth Thielke, Michael Sullivan, Jennifer Gaegler, Janet Manual, Jennifer R. Heinz, Ray Zisumbo, Diane C. Strollo, Candace Mueller, Irene Mahon, Brenda Delfosse, Carolyn M. Johnson, William E. Grizzle, Merideth Stanley, Sylvan Green, Pamela Harvey, Lindsay Richardson, Brenda K. Brewer, Philip Costello, Deanna Zapolski, John Worrell, Jeffrey G. Schragin, David S. Alberts, Edward L. Korn, Tamara Owens, Hank Brastater, Kay Mathiesen-Viergutz, Mark Broschinsky, Paul W. Spirn, Grace Isaacs, John S. Waltz, Mitch Goodsitt, Christi Newton-Foster, Sharlene Snowden, Barbara Voight, Gail Bizer, Kathy McDonough, William Huynh, Eduard Van Stam, Robert A. Carlson, Mike Florzyk, Paula M. Jacobs, Joan Fuller, Mauren Grunenwald, Ann Bangerter, Jacksonville, Adriane Andersen, Tess Thompson, Kenneth Nowers, Stephanie Helwi, Martin J. Edelman, Emmanuel Omoba, Rubenia Flores, Kevin T. White, Patrick W. Wolfe, Michael Milacek, Sharon Gard, Brandon B. Bigby, Cynthia H. McCollough, Andrew Burnside, Sheryl L. Ogden, Maisha Pollard, Thomas K. Pilgram, Sydney Laster, Claudia J. Kasales, Bruce W. Turnbull, Cheri Haselhuhn, Laura N. Myers, Jean Jacobsen, Melissa Love, Gavin D. Watt, Cheryl Love, Gerald F. Abbott, Susanne Kozakowski, Jerry L. Montague, Cynthia Hill, Neil F. O'Donnell, Anna Sear, Thomas M. Beck, Jean Wegner, Chrispina Wray, Edward M. Brown, Louise Ledbetter, Karen Bellware, Julie Moody, Noel Bahr, Matthew T. Freedman, Thomas Hensley, John E. Madewell, Leanne Hadfield, David R. Maffitt, Lisa Cottrell, John J. Warner, Deborah Graham, Krystal Arnold, Alejandra Reyes, Kristin Lieberman, Derek Omori, Donna Garland, Mike Burek, Mel Johnson, Judith Harkins, Martha Fronheiser, M. Y. M. Chen, Dawn Simmons, Kathleen Voight, Aaron O. Bungum, Marianne Rice, Lakeshia Murray, Tami Krpata, Donna Sammons, Leslie Kmetty, Catherine Duda, Carissa Krzeczkowski, Anne Nguyen, Richard H. Lane, Cynthia Mack, Loren C. Macey, Eddy Wicklander, Kelly McDaniel, Sue Zahradka, Hassan Bourija, Cristina Farkas, Jincy George, Renae Kiffmeyer, Wendell Christie, Catherine Engartner, John Crump, Mimi Kim, Carol Steinberg, Reginald F. Munden, Deb Kirby, Jo Ann Stetz, Barbara O'Brien, Sally Tenorio, Laura Multerer, Carlotta McCalister-Cross, Jessica Silva-Gietzen, Tamara Saunders, Harvey Glazer, Cam Vashel, Maria Oh, Rodkise Estell, Steven M. Moore, Tara Riley, Grant Izmirlian, D. Claire Anderson, James Burner, Steven Peace, Phil Hoffman, Angela Del Pino, Brian Irons, Carlos Jamis-Dow, John K. Lawlor, Edward F. Patz, Jay Afiat, Amber Barrow, Bawn M. Beno, Melissa S. Fritz, Lynn Coppage, Scott J. Sheltra, Tim Swan, Jerry Bergen, Charlie Fenton, Eric Deaton, Marilyn J. Siegel, Korinna Vigeant, Kerry Engber, Sarah Merrill, Buddy Williams, Kimberly Stryker, Bradley S. Snyder, Christina Romo, Andrea Hugill, Michael J. O'Shea, Linda White, Gail Fellows, Yasmeen Hafeez, Joe Woodside, Shauna Dave Scholl, Philip C. Prorok, Sharon Carmen, Kelly Hatton, Steven V. Marx, Sooah Kim, Robert Kobistek, Dawn Thomas, Lea Momongan, Chris Steward, Kari Bohman, Holly Bradford, Bradley S. Sabloff, Phillip Peterson, William C. Black, Lisa Pineda, James G. Ravenel, Karen Taylor, Beverly Trombley, Mona N. Fouad, Amber McDonald, Lauren J. Ramsay, Lisa Harmon, Jeffrey Geiger, David L. Spizarny, Jeffrey S. Klein, Xizeng Wu, Heather Tumberlinson, Joy Espiritu, Gina Varner, Dawn Fuehrer, Eric A. Hoffman, Sheila Moesinger, Nina Wadhwa, Steve King, Patricia Lavernick, Paola Spicker, Timothy R. Church, Cheryl Whistle, Sheila Greenup, Patricia Fantuz, Stephanie Levi, Peter Balkin, Mary E. Johnson, Johanna Ziegler, Susan Hoffman, Kathy L. Clingan, Craig Kuhlka, Maria Marchese, Lawrence F Cohen, Cylen Javidan-Nejad, Wilbur A. Franklin, Kevin J. Leonard, Tim A. Parritt, Jade Quijano, Kathleen Poler, Jennifer Rosenbaum, Xiuli Zhang, Christine Brown, Terri David-Schlegel, Susan M. Peterson, James R. Jett, Kenneth W. Clark, Edward P. Gelmann, Arthur Migo, Patricia Fox, Lori Hamm, Janie McMahon, Darlene Guillette, Robert C. Young, Patty Beckmann, Jerome Jones, Nikki Jablonsky, Roberta Yoffie, Heather L. Bradley, Darlene Higgins, Francine L. Jacobson, Christine B. Berg, Mark Bramwitt, Constantine N. Petrochko, Karen Stokes, Jennifer Rowe, Kathy McKeeta-Frobeck, Brenda Sleasman, Courtney Bell, Dave Tripp, Saundra S. Buys, Susan Walsh, Jo Rean D. Sicks, Richard G. Barr, Kirk Midkiff, Tom Caldwell, Elisabeth A. Grady, Subbarao Inampudi, Marilyn Calulot, Paul A. Kvale, Alice DuChateau, Kathy Berreth, Ruth Holdener, Katie Kuenhold, Thomas E. Warfel, David P. Naidich, Mandie Leming, Fraser Wilton, Leanne Franceshelli, Kathleen McMurtrie, Elaine Bowman, Donald F. Bittner, Helen Kaemmerer, Merri Mullennix, Adelheid Lowery, Andrew Karellas, Jenny Hirschy, Kate Naughton, Ashley B. Long, Kristin M. Gerndt, Kathleen Young, Richard M. Schwartzstein, Wendy Smith, Joseph Aisner, Shane Ball, Kathleen Krach, Cathy Mueller, Virginia May, Christopher Blue, Marsha Lawrence, Ronald S. Kuzo, Colleen McGuire, Alisha Moore, Sara Cantrell, Christie Leary, Pamela Allen, Maryann Trotta, Clifford Caughman, Peggy J. Gocala, Brian Mullen, Janan Alkilidar, Maryann Duggan, Lin Mueller, Alesis Nieves, Fenghai Duan, Frederick Olson, Edwin G. Williams, Jo Ann Hall Sky, Grant Izmirilian, Peggy Joyce, Judy Preston, Cristine Juul, Julianne Falcone, Bruce Neilson, Fla Lisa Beagle, Beth Evans, Jamie Mood, Janet Bishop, Jean Tsukamoto, Vivien Gardner, Gillian Devereux, Minesh Patel, Sally Fraki, Celia Stolin, Ami Lyn Taplin, Stephenie Johnson, Saeed Matinkhah, Jenna Bradford, Sanjeev Bhalla, Charles Jackson, Julie Haglage, Darlene R. Fleming, Allie M. Bell, Paul A. Bunn, Gail Orvis, Andrew J. Bierhals, Julie Ngo, Belores K. Prudoehl, Elaine N. Daniel, Peggy Olson, Paul F. Pinsky, Glenna M. Fehrmann, Aras Acemgil, Andrea Hamilton-Foss, Leeta Grayson, Smita Patel, Scott Emerson, Carl J. Zylak, James R. Maxwell, Jennifer Fleischer, Suzanne Smith, Jacqueline R. Sheeran, Alan Williams, Scott Gaerte, John Fletcher, Sonya Clark, Nancy Gankiewicz, Stuart S. Sagel, Jason Spaulding, Nancy E. Hanson, Nicole Fields, Richard D. Nawfel, Dinakar Gopalakrishnan, Margaret Oechsli, Susan Wenmoth, Isabelle Forter, Elizabeth Morrell, Jessica Rider, Letitia Clark, Michael Woo, Cynthia A. Brown, Camille Mueller, Mark T. Dransfield, Lois M. Roberts, Anne Randall, Eduard J. Gamito, Carrie O'Brien, Carolyn Palazzolo, Julie Schach, Robert Falk, Melissa Hudson, Jennifer Garcia Livingston, Cynthia L. Andrist, Tammy Fox, Elliott Drake, Tanya Zeiger, Renee Metz, Kevin Thomas, Neha Kumar, Elizabeth Couch, Beborah Bay, Mei Hsiu Chen, Jason Bronfman, Philip Dennis, Deb Engelhard, Pamela McBride, Daniel Kimball, Amy Haas, Pamela M. Mazuerk, Marlea Osterhout, Venetia Cooke, Tina Taylor, Amy St.Claire, Joe Hughes, Becky McElsain, Beverly Brittain, Michele Adkinson, Paige Beck, Martha Maineiro, Paula R. Beerman, Jackie Seivert, Mary M. Pollock, Donald Corle, Tina Herron, Marcella Petruzzi, Natalie F. Scully, Kenneth A. Coleman, Jennifer Yang, Debra Loria, Wendy Moss, Alan Brisendine, Cheryl M. Lewis, Dalphany Blalock, Lonni Schultz, Douglas Bashford, Nora Szabo, David Shea, Amanda Devore, Karen Schleip, Judy Netzer, Barry Clot, Gerald M. Mulligan, Nancy E. Krieger Black, David Schultz, Jim Pool, Craig E. Leymaster, Kathryn Rabanal, Kay Bohn, Tara Berg, Marisol Furlong, Stacey Mitchell, Donna Biracree, Laura Jones, Cassie Olson, Robin Stewart, Jeremy Pierce, Marilyn Bruger, Valene Kennedy, Stephanie Davis, Colin O'Donnell, Glenn A. Tung, Shannon Wright, William Lake, Sharon Jones, Vincent Girardi, Brad Benjamin, Veenu Harjani, Drew A. Torigian, Kevin Edelman, Sue Frederickson, Paul E. Smart, Michelle Wann Haynes, D S Gierada, Glenn Fletcher, Rosalie Ronan, Patricia Ann Street, Eleace Eldridge-Smith, Lynly Wilcox, Cindy Lewis-Burke, La Tonja Davis, Rachel Black Thomas, Dawn Shone, Evangeline Griesemer, Tim Budd, Lindsey Dymond, Marlene Semansky, Amy Rueth, Constantine Gatsonis, Kay H. Vydareny, Usha Singh, Amy Lita Evangelista, Angelica C. Barrett, Bethany Pitino, Shirley Wachholz, Angela M. Williams, Sandra Fiarman, Karen Luttrop, David Chellini, Michael Bradley, Helen Fink, Aaron Zirbes, Roger Inatomi, Joon K. Lee, Heather Bishop Blake, Lisa Woodard, Craig Hritz, Sarah Neff, Aine Marie Kelly, Deborah Harbison, Baigalmaa Yondonsambuu, Amy Lloyd, Christine Gjertson, Erin Cunningham, Angelee Mean, June Morfit, Ping Hu, William Thomas, Jazman Brooke, Paul Marcus, Jeremy Gorelick, Erin Lange, William Stanford, Denise R. Aberle, Lena Glick, Annabelle Lee, Ian Malcomb, Deanna L. Miller, Mary Mesnard, Jacqueline Jackson, Jhenny Hernandez, Desiree E. Morgan, Howard I. Jolies, Jacquie Marietta, Teresa Lanning, Debra Rempinski, Amanda C. Davis, Karen Mathews Batton, Mahadevappa Mahesh, Erik Wilson, Deana Nelson, Sharan L. Campleman, William Manor, Julie Sears, Howard Mann, E. David Crawford, Carl Krinopol, Greg Gambill, Margo Cousins, Rex C. Yung, Sangeeta Tekchandani, Thomas Vahey, Ann D. McGinnis, Kimberly Nolan, Kaylene Crawford, Kelli P. Rockwell, Dana Roeshe, Fred W. Prior, Kari Ranae Kramer, Heidi Nordstrom, Frank Stahan, Shawn Sams, Cherie Baiton, Joy Tani, Thomas J. Watson, Angela Cosas, Diane Kowalik, Pritha Dalal, Ann Jolly, Jeanine Wade, Laura Bailey, Julie Varner, Glen K. Nyborg, Christopher Toyn, David Gemmel, Susanna N. Dyer, Laurie Amendolare, Mary Ellen Frebes, Judy Ho, Adele Perryman, John Keller, D. Sullivan, George Mahoney, Scott Cupp, Linda L. Welch, Peter Greenwald, Robert Sole, Marcello Grigolo, Caroline Chiles, Patricia Sheridan, Deborah M. Chewar, Vijayasri Narayanaswami, Susan Blackwell, Suzanne B. Lenz, Alphonso Dial, Melvin Tockman, Carolyn Hill, John Stubblefield, Catherine E. Smith, Judith Lobaugh, Rosa M. Medina, Jackie Meier, Nandita Bhattacharjee, Robert Tokarz, Lisa Clement, Nancy Caird, Cindy Masiejczyk, Patricia Shwarts, Laura Springhetti, Sandra Schornak-Curtis, Edwin F. Donnelly, Patricia Tesch, Laurie Rathmell, Pamela K. Woodard, Edward A. Sausville, David R. Pickens, Kylee Hansen, Paulette Williams, Barbara Ferris, Rachel L. McCall, Nicole M. Carmichael, Dawn Whistler, Ramachandra Chanapatna, Glynis Marsh, Mary Wiseman, Tony DeAngelis, L. Heather, Vicki Prayer, Robin Laura, Priscilla Bland, Gregory W. Gladish, Amy Garrett, Kelly McNulty, Daniel J. Pluta, Mylene T. Truong, Serelda Young, Crista Cimis, Gordon Jacob Sen, Rhonda Rosario, Anthony B. Miller, Edward Hunt, Juanita Helms, Jill K. Bronson, Jeff Yates, Ginette D. Turgeon, Bo Lu, Nancy Fredericks, Pam Senn, Ryan Pena, Hakan Sahin, Mary Lynn Steele, Jill E. Cordes, Noel Maddy, R. Adam DeBaugh, Hope Hooks, Zipporah Lewis, Robert L. Berger, Shani Harris, Natalie Gray, Jennifer Kasecamp, Elizabeth King, Jacinta Mattingly, Hrudaya Nath, Kathy Torrence, Christine Cole Johnson, Sara Mc Clellan, Kalin Albertsen, Kim Sprenger, Ryan Norton, Jody Wietharn Kristopher, Linda Warren, Byung Choi, Casey O'Quinn, Mark K. Haron, Chris J. Jennings, Karen Robinson, Joan Molton, Dorothy Hastings, Robert I. Garver, Christopher J. Cangelosi, Jeannette Lynch, Peter Ohan, Angela Campbell, Dawn Mead, Miriam Galbraith, Divine Hartwell, Natalya Portnov, Gene L. Colice, Andetta R. Hunsaker, Analisa Somoza, Todd Risa, Daniel C. Sullivan, Karthikeyan Meganathan, Tammy DeCoste, Peter Zamora, Richard M. Fagerstrom, Iiana Gareen, Phyllis J. Walters, Barbara L. Carter, Alem Mulugeta, Rob Bowman, Kavita Garg, Andrea Franco, Mary Adams Zafar Awan, Edward Reed Smith, Rachel Phillips, Michelle Aganon-Acheta, Fred R. Hirsch, Peter Jenkins, Pamela Taybus, Joy Knowles, Karen M. Horton, Cheryl Spoutz-Ryan, Sarah Landes, William G. Hocking, Laura B. Schroeder, Erini Makariou, Jered Sieren, Kaylene Evans, Erin Nekervis, Brenda Polding, Tonda Robinson, Joel L. Weissfeld, Terry J. Sackett, Michael F. McNitt-Gray, Leslie Dobson, Raymond Weatherby, Randell Kruger, Revathy B. Iyer, Mary Krisk, Anthony Levering, Susan Collins, Alison Schmidt, William M. Hanson, Patricia Schuler, Karen Glanz, Morgan Ford, Beatrice Trotman-Bickenson, Richard Guzman, Paul Koppel, Judith K. Amorosa, Meredith Slear, Dayna Love, Carol Vaughn, Kellyn Adams, Celeste Monje, Garry Morrison, Sherri Mesquita, Paul Cronin, Tony Blake, Constance Elbon-Copp, Robert A. Clark, Felix Mestas, Erich Allman, Armen Markarian, Cheryl Souza, Karen O’Toole, Elliot K. Fishman, Karen Augustine, Jane Hill, Bonnie Kwit, Ralph Drosten, Susan Foley, Stacy E. Smith, Angie Bailey, Jennifer Bishop Kaufmann, Shelly Meese, Phillip M. Boiselle, Howard Morrow, Thomas D. Hinke, Barry Edelstein, Erin Schuler, William C. Bailey, Donna Letizia, David S. Gierada, Frederick J. Larke, Robin Haverman, Sarah Baum, Sally Hurst, Richard L. Morin, Ben Dickstein, William Russell, J. Anthony Seibert, Sophia Sabina, Mary Alyce Riley, Michael A. Taylor, Katherine BeAngelis, Robert A. Hawkins, Fernando R. Gutierrez, Amie Welch, Heather Lancor, George Armah, James Blaine, Eric Henricks, Joel Dunnington, Carole Walker, Laura Motley, Melody Kolich, Bruce J. Hillman, David W. Sturges, Mindy Lofthouse, Amy Warren, Michael Black, Mark Kolich, Lisa A. Holloway, Shannon M. Pretzel, Susan Shannon, Yassminda Harts, Dallas Sorrel, Lance A. Yokochi, Diana Wisler, Arthur Sandy, Roberta Clune, Shirley Terrian, Shalonda Manning, Bradley Willcox, Thomas J. Payne, James L. Tatum, Dale Brawner, Sandy Morales, Rodolfo C. Morice, Amy Vieth, Emily Jewitt, Chelsea O'Carroll, Theresa C. McLoud, John E. Langenfeld, Chris H. Cagnon, Lisa B. Hinshaw, Gena Kucera, Helena R. Richter, Drew Torigian, June McSwain, Courtney Eysmans, Vinis Salazar, David Spizarny, Mary Kelly-Truran, Mark Whitty, Henry Albano, Connie L. Sathre, William R. Geiser, Barnett S. Kramer, Marianna Gustitis, Gordon C. Jones, Neil E. Caporaso, Timothy Welsh, Roger Tischner, Ana Maria Mendez, Dominick A. Antico, Cathy L. Bornhorst, Carla Chadwell, Stephanie Pawlak, Kelli M. West, Joe V. Selby, Randall Kruger, Jodi Hildestad, Elaine Freesmeier, Nicole Rivas, Andrew Goodman, Naima Vera-Gonzalez, Stuart Lutzker, Eric M. Hart, Melanie Yeh, Shane Sorrell, Deb Multerer, Sharon Jacoby, Debbie Gembala, Elizabeth Fleming, Myrle Johnson, Michael J. Flynn, Frank Tabrah, Martin L. Schwartz, Deanna Mandley, Brad Siga, Guillermo Marquez, Jeffrey Koford, Victoria Jenkins, Janice Pitts, Constantine A. Gatsonis, Natalie Baptiste, Edith M. Marom, Gina Sammons, Anne Burrough, Martha Ramirez, Jack Cahill, Carl Jaffe, Linda Heinrichs, Aura Cole, Paul Rust, Alon Coppens, Gregg Hamm, Lisa Conklin, Kathleen A. Robbins, Carleaner Williams, Gwen Chalom, Winston Sterling, Colleen Hudak, Lea Matous, Ella A. Kazerooni, Denise Kriescher, David A. Lynch, Liz Bolan, Jacob Wolf, Jonathan G. Goldin, Roberta Quinn, L. A. Schneider, Kathleen A. Murray, Erica Sturgeon, Jennifer Avrin, Michelle T. Biringer, Mark Hinson, Cynthia Reiners, Brian Chin, Amy Brunst, Ann M. Lambrecht, Katherine Lohmann, Jennifer Bacon, Ulander Giles, Diane Shepherd, William T. Corey, Timothy Cosgrove, Lana C. Walters, Nancy Kadish, Hilary C. Nosker, Christine D. Berg, Thomas Payne, Jackie Becker, Kanistha Sookpisal, Lyn Seguin, Todd R. Hazelton, Roy Adaniya, James Fisher, Annmarie Walsh, Shirleen Hyun, Laura Stark, Kenneth Hansen, Carolyn Nelson, Martin Tammemagi, Mary A. Wolfsberger, Barry H. Gross, Valentina Ortico, Marge Watry, Jeff Childs, Gabe Herron, Loretta Thorpe, Lisa Damon, Evanthia Papadopoulos, Denise Moline, Voula E. Christopoulos, John D. Minna, Tony Jones, Mitchell Machtay, Michael Plunkett, Melissa Laughren, Luis Zagarra, Adam Leming, Eda Ordonez, Chris Howell, Marissa Peters, Wendy Mosiman, Joanne Gerber, Alfonso Lorenzo, Barbara L. McComb, Laura Hill, Gale Christensen, Hanna Comer, Carmen Guzman, Kathy Taylor, Misty Oviatt, Malcolm King, Lily Stone, Rex Welsh, Bernadette Pennetta, Cristina Raver, Jan E. Hyder, Stephanie Clabo, Peggy Lau, Jacqueline Fearon, Patricia Pangburn, Pamela Dow, William K. Evans, Victor De Caravalho, Mike Wirth, Brooke Johnson, Meridith Blevins, Lisa H. Gren, Sharon L. Kurjan, James P. Evans, Kirk E. Smith, Donna King, John A. Worrell, Mindy S. Geisser, Philip F. Judy, Richard Barr, Sue Misko, Stanley R. Phillips, Jillian Nickel, Christine M. McKey, Joe Austin, Donna Hartfeil, Laura Young, Shovonna White, Alexis K. Potemkin, Anthony Boulos, Tawny Martin, Karen Kofka, Heather McLaughlin, Matthew K. Siemionko, Melissa Houston, Angela Lee Rowley, Adys Fernandez, Murray Backer, Jagdish Singh, Mary Weston, Nancy Payte, Charles Apgar, John K. Gohagan, Jeff Fairbanks, Wylie Burke, David Chi, Michael Nahill, Kevin DeMarco, Karen Patella, Beverly Rozanok, Carol M. Moser, Nicole Matetic Mac, Karen Boyle, Dinah Lorenzo, Elanor Adkins, Phyllis Olsson, Amanda M. Adams, Sujaya Rao, K.E. Jones, Polly Kay, D. Lynn Werner, John B. Weaver, Sally Anne Kopesec, Jennifer Frye, Victoria Chun, Cathy Francow, Cheri Whiton, Jo Ann Nevilles, Andrew Bodd, Barbara Galen, Sabrina Chen, Cindy Cyphert, Stephen M. Moore, Petra J. Lewis, Shanna Nichols, Mareie Walters, Thea Palmer Zimmerman, Warren B. Gefter, Peter Dubbs, Ann Reinert, Holly Washburn, Renee MacDonald, Boleyn R. Andrist, Dianalyn M. Evans, Marvin Flores, Tricia Adrales-Bentz, Claudine Isaacs, Regina C. MacDougall, Greg M. Silverman, Nichoie Cadez, Lynne Bradford, Rochelle Williams, Angela M. McLaughlin, Ellen Sandberg, Cheryl Crozier, Robert Mayer, Richard P. Remitz, Sheron Bube, Leroy Riley, Vish Iyer, Sophie Breer, Stephen Baylin, Anna Boyle, Shannon Williams, Kristen Keating, Martin M. Oken, Gerald L. Andriole, Bruce E. Hubler, Eric T. Goodman, David Engelhart, Bonna Au, Brianne Whittaker, Tricia Hoffa, Eng Brown, Tammy Wolfsohn, Denise L. Foster, Barry H. Cohen, Linda Galocy, Matthew T. Bee, Jacqueline Matuza, Leslie Henry, Katherine Meagher, Mona Fouad, Beth McLellan, Troy Cook, John Sheflin, Lilian Villaruz, Marcella Moore, Brandy Mack-Pipkin, Vanessa Graves, Ryan Weyls, William T. Herbick, Geoffrey McLennan, Lynn Hoese, Janise Webb, Terrie Kitchner, Michele Lee, Robert T. Greenlee, Charles C. Matthews, Nicole Spiese, Jeffrey Heffernon, Dianna D. Cody, Patricia Blair, Kathy Garrett, Michael A. Sullivan, and Loretta Granger

Subjects
Oncology, medicine.medical_specialty, business.industry, Mortality rate, medicine.disease, law.invention, Quality-adjusted life year, Randomized controlled trial, law, Internal medicine, medicine, Radiology, Nuclear Medicine and imaging, National Lung Screening Trial, Radiology, Overdiagnosis, business, Lung cancer, Lung cancer screening, Mass screening
Abstract

The National Lung Screening Trial (NLST) is a randomized multicenter study comparing low-dose helical computed tomography (CT) with chest radiography in the screening of older current and former heavy smokers for early detection of lung cancer, which is the leading cause of cancer-related death in the United States. Five-year survival rates approach 70% with surgical resection of stage IA disease; however, more than 75% of individuals have incurable locally advanced or metastatic disease, the latter having a 5-year survival of less than 5%. It is plausible that treatment should be more effective and the likelihood of death decreased if asymptomatic lung cancer is detected through screening early enough in its preclinical phase. For these reasons, there is intense interest and intuitive appeal in lung cancer screening with low-dose CT. The use of survival as the determinant of screening effectiveness is, however, confounded by the well-described biases of lead time, length, and overdiagnosis. Despite previous attempts, no test has been shown to reduce lung cancer mortality, an endpoint that circumvents screening biases and provides a definitive measure of benefit when assessed in a randomized controlled trial that enables comparison of mortality rates between screened individuals and a control group that does not undergo the screening intervention of interest. The NLST is such a trial. The rationale for and design of the NLST are presented.

Published
2011
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12. Real-Time DNA Sequencing from Single Polymerase Molecules

Author

Jeremy Gray, Mark Trulson, Patrick Marks, John Dixon, Ravindra V. Dalal, Fred Christians, Adrian Fehr, Jon M. Sorenson, Stephen Turner, Alfred Gaertner, Sonya Clark, Geoff Otto, Gregory J. Kearns, John Lyle, Alex DeWinter, Brad Bettman, Ronald Kuse, Primo Baybayan, Steven Lin, Denis Zaccarin, John Vieceli, Joy Roy, Cheryl Heiner, David R. Rank, Kevin Travers, Robert Sebra, Mathieu Foquet, Thang Pham, Dawn Wu, Keith Bjornson, Michael Phillips, Arkadiusz Bibillo, Bidhan Chaudhuri, Gene Shen, Alicia Yang, Mark Maxham, Peter Zhao, Khai Luong, Paul Hardenbol, Insil Park, Jonas Korlach, Paul Lundquist, Jeffrey Wegener, Kevin Hester, David P. Holden, Paul Peluso, Congcong Ma, Frank Zhong, Yves Lacroix, Austin B. Tomaney, Devon Murphy, John Eid, Xiangxu Kong, and Ronald L. Cicero

Subjects
DNA nanoball sequencing, Deoxyribonucleotides, DNA, Single-Stranded, DNA-Directed DNA Polymerase, Sequencing by hybridization, Consensus Sequence, Polymerase, Fluorescent Dyes, Multidisciplinary, DNA clamp, Base Sequence, biology, Oligonucleotide, Multiple displacement amplification, DNA, Sequence Analysis, DNA, Enzymes, Immobilized, Molecular biology, Nanostructures, Sequencing by ligation, Kinetics, Spectrometry, Fluorescence, biology.protein, Biophysics, DNA, Circular, Single molecule real time sequencing
Abstract

We present single-molecule, real-time sequencing data obtained from a DNA polymerase performing uninterrupted template-directed synthesis using four distinguishable fluorescently labeled deoxyribonucleoside triphosphates (dNTPs). We detected the temporal order of their enzymatic incorporation into a growing DNA strand with zero-mode waveguide nanostructure arrays, which provide optical observation volume confinement and enable parallel, simultaneous detection of thousands of single-molecule sequencing reactions. Conjugation of fluorophores to the terminal phosphate moiety of the dNTPs allows continuous observation of DNA synthesis over thousands of bases without steric hindrance. The data report directly on polymerase dynamics, revealing distinct polymerization states and pause sites corresponding to DNA secondary structure. Sequence data were aligned with the known reference sequence to assay biophysical parameters of polymerization for each template position. Consensus sequences were generated from the single-molecule reads at 15-fold coverage, showing a median accuracy of 99.3%, with no systematic error beyond fluorophore-dependent error rates.

Published
2009
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13. Long, Processive Enzymatic DNA Synthesis Using 100% Dye-Labeled Terminal Phosphate-Linked Nucleotides

Author

Paul Peluso, Arek Bibillo, Insil Park, Jonas Korlach, Sonya Clark, Jeffrey Wegener, Thang Pham, Geoff Otto, and Stephen Turner

Subjects
Deoxyribonucleoside triphosphate, sequencing applications, DNA polymerase, Base pair, Biochemistry, Genetics, Nucleotide, Fluorescent Dyes, chemistry.chemical_classification, DNA clamp, Molecular Structure, biology, Nucleotides, Chemistry, DNA replication, DNA, General Medicine, dNTP, Kinetics, biology.protein, Molecular Medicine, Original Article, fluorescence, DNA polymerase I, Dideoxynucleotides, Single molecule real time sequencing
Abstract

We demonstrate the efficient synthesis of DNA with complete replacement of the four deoxyribonucleoside triphosphate (dNTP) substrates with nucleotides carrying fluorescent labels. A different, spectrally separable fluorescent dye suitable for single molecule fluorescence detection was conjugated to each of the four dNTPs via linkage to the terminal phosphate. Using these modified nucleotides, DNA synthesis by phi 29 DNA polymerase was observed to be processive for products thousands of bases in length, with labeled nucleotide affinities and DNA polymerization rates approaching unmodified dNTP levels. Results presented here show the compatibility of these nucleotides for single-molecule, real-time DNA sequencing applications.

Published
2008
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15. PHYSICAL CHARACTERIZATION OF ALLIINASE, THE FLAVOR GENERATING ENZYME IN ONIONS

Author

Sonya Clark, Martin L. Shaw, Dale Every, and Jane E. Lancaster

Subjects
Pharmacology, Gene isoform, Glycosylation, biology, Molecular mass, Chemistry, Protein subunit, Biophysics, Protein primary structure, Fast protein liquid chromatography, Cell Biology, chemistry.chemical_compound, Biochemistry, Alliinase, Complementary DNA, biology.protein, Food Science
Abstract

The characteristic flavor of onions occurs when the enzyme alliinase (EC 4.4.1.4) hydrolyses the S-alk(en)yl-L-cysteine sulfoxides to form sulfur containing volatiles, pyruvate and ammonia. In this paper we have new evidence on the molecular mass of native onion alliinase, its subunit size and its relationship to the predicted size from translation of the gene. Alliinase from bulbs was purified to homogeneity. Two isoforms of alliinase were detected on ion-exchange chromatography. Both isoforms were analyzed by gel filtration FPLC. Alliinase activity was detected at molecular masses of 59, 166 kDa and 59, 170 and 393 kDa for isoforms 1 and 2, respectively. When analyzed by SDS-PAGE subunit sizes were determined to be 53.3 kDa for isoform 1 and 53.3 and 51.6 kDa for isoform 2. Amino terminal sequencing of both subunits confirmed they were alliinase. Alliinase was deglycosylated and the subunits migrated as a single band indicating a common protein backbone and varying glycosylation. Thus native alliinase is a monomer, trimer and multimer of the subunits. Molecular analysis of alliinase cDNA indicated that two genes and thus two protein subunits were expressed in onion bulb tissue. Translation is likely to have initiated from the third methionine of the leader sequence of the cDNA to give a protein backbone of 53.5 kDa in gradient SDS-PAGE. The functional significance of heterogenous subunits, from different genes, and multimeric native alliinase remains unknown.

Published
1998
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16. A folded protein can be transported across the chloroplast envelope and thylakoid membranes

Author

Steven M. Theg and Sonya Clark

Subjects
Protein Folding, Chloroplasts, Photosystem II, Recombinant Fusion Proteins, Photosynthetic Reaction Center Complex Proteins, Gene Expression, Biology, Peptides, Cyclic, Chloroplast membrane, Twin-arginine translocation pathway, Aprotinin, Animals, Molecular Biology, Photosystem II Protein Complex, food and beverages, Biological Transport, Intracellular Membranes, Cell Biology, Hydrogen-Ion Concentration, Membrane transport, Transmembrane protein, Chloroplast, Cross-Linking Reagents, Biochemistry, Thylakoid, Biophysics, Cattle, Protein folding, Research Article
Abstract

Many thylakoid lumenal proteins are nuclear encoded, cytosolically synthesized, and reach their functional location after posttranslational targeting across two chloroplast envelope membranes and the thylakoid membrane via proteinaceous transport systems. To study whether these transmembrane transport machineries can translocate folded structures, we overexpressed the 17-kDa subunit of the oxygen-evolving complex of photosystem II (prOE17) that had been modified to contain a unique C-terminal cysteine. This allowed us to chemically link a terminal 6.5-kDa bovine pancreatic trypsin inhibitor (BPTI) moiety to prOE17 to create the chimeric protein prOE17-BPTI. Redox reagents and an irreversible sulfhydryl-specific cross-linker, bis-maleimidohexane, were used to manipulate the structure of BPTI. Import of prOE17-BPTI into isolated chloroplasts and thylakoids demonstrates that the small tightly folded BPTI domain is carried across both the chloroplast envelopes and the delta pH-dependent transmembrane transporter of the thylakoid membrane when linked to the correctly targeted OE17 precursor. Transport proceeded even when the BPTI moiety was internally cross-linked into a protease-resistant form. These data indicate that unfolding is not a ubiquitous requirement for protein translocation and that at least some domains of targeted proteins can maintain a nonlinear structure during their translocation into and within chloroplasts.

Published
1997
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17. Metabolic engineering of Corynebacterium glutamicum for anthocyanin production

Author

Jian Zha, Ying Zang, Matthew Mattozzi, Jens Plassmeier, Mamta Gupta, Xia Wu, Sonya Clarkson, and Mattheos A. G. Koffas

Subjects
Anthocyanin, Corynebacterium glutamicum, Catechin, Flavonoid, UDP-glucose, Microbiology, QR1-502
Abstract

Abstract Background Anthocyanins such as cyanidin 3-O-glucoside (C3G) have wide applications in industry as food colorants. Their current production heavily relies on extraction from plant tissues. Development of a sustainable method to produce anthocyanins is of considerable interest for industrial use. Previously, E. coli-based microbial production of anthocyanins has been investigated extensively. However, safety concerns on E. coli call for the adoption of a safe production host. In the present study, a GRAS bacterium, Corynebacterium glutamicum, was introduced as the host strain to synthesize C3G. We adopted stepwise metabolic engineering strategies to improve the production titer of C3G. Results Anthocyanidin synthase (ANS) from Petunia hybrida and 3-O-glucosyltransferase (3GT) from Arabidopsis thaliana were coexpressed in C. glutamicum ATCC 13032 to drive the conversion from catechin to C3G. Optimized expression of ANS and 3GT improved the C3G titer by 1- to 15-fold. Further process optimization and improvement of UDP-glucose availability led to ~ 40 mg/L C3G production, representing a > 100-fold titer increase compared to production in the un-engineered, un-optimized starting strain. Conclusions For the first time, we successfully achieved the production of the specialty anthocyanin C3G from the comparatively inexpensive flavonoid precursor catechin in C. glutamicum. This study opens up more possibility of C. glutamicum as a host microbe for the biosynthesis of useful and value-added natural compounds.

Published
2018
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