Biotechnology Commons^™

Arizona Clinical Research Workforce Survey, Cris E. Wells, Julie A. Robbins, Gaye Luna

Online Journal for Workforce Education and Development

Objective: We characterize the current clinical research workforce in Arizona with the goal of identifying possible educational training gaps for the future workforce. Background: The science community, government entities, educational institutions, and community programs continue to advocate for additional funding and resources to educate the bioscience workforce, of which clinical research is a part. However, there is little evidence that additional training and/or education is needed within the clinical research profession. Method: We utilized an electronic survey tool that was designed to solicit workforce data from clinical research professionals actively working or seeking employment in the field of clinical research …

Measuring Radiation Exposure In Human Blood Using Gene Expression, Krystal Naranjo, Melissa Bentley, Harsha Konery, Matthew Coleman

STAR Program Research Presentations

Mammalian cells are known to express genes that are associated with repairing damaged DNA. The transcript CDKN1A is one of several cell cycle regulator genes expressed in response to cell damage by ionizing radiation (IR). In this study, male and female lymphocytes; previously exposed ex vivo to IR, were used to demonstrate linear gene expression responses that may vary between genders. We used qRT-PCR to generate response curves for CDKN1A. No differences were identified for the endogenous control gene GAPDH. CDKN1A expression demonstrated average fold changes well above three fold for three of the four healthy patient donors at 24 …

The Effects Of Electrolyte Chemical Composition, Concentration And Ph On The Electrochemical Redox Reactions, Sea Choi, Xiao-Ying Yu

STAR Program Research Presentations

Electrolyte composition, concentration, pH, ionic strength, and temperature have long been known to play a role in the interface of the electrode-electrolyte. However, detailed studies are not sufficient. Cyclic voltammetry (CV) is a widely used electrochemical technique for obtaining qualitative information in electroanalysis. CV is often the first experiment performed in an electrode surface in an electrochemical study. It measures the number of electrons transferred and intermediates formed during oxidation-reduction (redox) reactions by observing its current, intensity and position of peak potential with applied voltage. This is accomplished by using a three-electrode system including working electrode (WE), counter electrode (CE) …

Physiological And Transcriptional Memory In Guard Cells During Repetitive Dehydration Stress, Laetitia Virlouvet, Michael E. Fromm

Nebraska Center for Biotechnology: Faculty and Staff Publications

Arabidopsis plants subjected to a daily dehydration stress and watered recovery cycle display physiological and transcriptional stress memory. Previously stressed plants have stomatal apertures that remain partially closed during a watered recovery period, facilitating reduced transpiration during a subsequent dehydration stress. Guard cells (GCs) display transcriptional memory that is similar to that in leaf tissues for some genes, but display GC-specific transcriptional memory for other genes. The rate-limiting abscisic acid (ABA) biosynthetic genes NINE-CIS-EPOXYCAROTENOID DIOXYGENASE 3 (NCED3) and ALDEHYDE OXIDASE 3 (AAO3) are expressed at much higher levels in GCs, particularly during the watered recovery interval, relative to their low …

The Pseudomonas Syringae Type Iii Effector Hopd1 Suppresses Effector-Triggered Immunity, Localizes To The Endoplasmic Reticulum, And Targets The Arabidopsis Transcription Factor Ntl9, Anna Block, Tania Y. Toruno, Christian G. Elowsky, Chi Zhang, Jens Steinbrenner, Jim Beynon, James R. Alfano

Nebraska Center for Biotechnology: Faculty and Staff Publications

Pseudomonas syringae type III effectors are known to suppress plant immunity to promote bacterial virulence. However, the activities and targets of these effectors are not well understood. We used genetic, molecular, and cell biology methods to characterize the activities, localization, and target of the HopD1 type III effector in Arabidopsis. HopD1 contributes to P. syringae virulence in Arabidopsis and reduces effector-triggered immunity (ETI) responses but not pathogen-associated molecular pattern-triggered immunity (PTI) responses. Plants expressing HopD1 supported increased growth of ETI-inducing P. syringae strains compared with wild-type Arabidopsis. We show that HopD1 interacts with the membrane-tethered Arabidopsis transcription factor NTL9 and …

Book Review Of Dane Kennedy, The Last Blank Spaces: Exploring Africa And Australia, Adrian S. Wisnicki

Department of Biochemistry: Faculty Publications

Mid-nineteenth-century British exploration, particularly the stories of the “heroic” individuals who carried out this exploration, remains a topic of worldwide interest, as most recently evidenced by the many events in Britain and Africa celebrating the 2013 bicentenary of David Livingstone’s birth. Dane Kennedy’s intriguing study takes issue with such readings of the historical record by foregrounding an epistemological tension that lies at the heart of nineteenthcentury British exploration discourse and practice. The book examines the collision—as it played out in the exploration of African and Australia—between metropolitan scientific protocols and non-Western quotidian realities. Nineteenth-century explorers, argues Kennedy, left home with …

Stress Adaptation In A Pathogenic Fungus, Alistair J. P. Brown, Susan Budge, Despoina Kaloriti, Anna Tillmann, Mette D. Jacobsen, Zhikang Yin, Iuliana V. Ene, Iryna Bohovych, Doblin Sandai, Stavroula Kastora, Joanna Potrykus, Elizabeth R. Ballou, Delma S. Childers, Shahida Shahana, Michelle D. Leach

Department of Biochemistry: Faculty Publications

Candida albicans is a major fungal pathogen of humans. This yeast is carried by many individuals as a harmless commensal, but when immune defences are perturbed it causes mucosal infections (thrush). Additionally, when the immune system becomes severely compromised, C. albicans often causes life-threatening systemic infections. A battery of virulence factors and fitness attributes promote the pathogenicity of C. albicans. Fitness attributes include robust responses to local environmental stresses, the inactivation of which attenuates virulence. Stress signalling pathways in C. albicans include evolutionarily conserved modules. However, there has been rewiring of some stress regulatory circuitry such that the roles of …

Xiap Antagonist Embelin Inhibited Proliferation Of Cholangiocarcinoma Cells, Cody J. Wehrkamp, Ashley R. Gutwein, Sathish Kumar Natarajan, Mary Anne Phillippi, Justin L. Mott

Department of Biochemistry: Faculty Publications

Cholangiocarcinoma cells are dependent on antiapoptotic signaling for survival and resistance to death stimuli. Recent mechanistic studies have revealed that increased cellular expression of the E3 ubiquitin-protein ligase X-linked inhibitor of apoptosis (XIAP) impairs TRAIL- and chemotherapy-induced cytotoxicity, promoting survival of cholangiocarcinoma cells. This study was undertaken to determine if pharmacologic antagonism of XIAP protein was sufficient to sensitize cholangiocarcinoma cells to cell death. We employed malignant cholangiocarcinoma cell lines and used embelin to antagonize XIAP protein. Embelin treatment resulted in decreased XIAP protein levels by 8 hours of treatment with maximal effect at 16 hours in KMCH and Mz-ChA-1 …

Nudging The Fda, W. Nicholson Price Ii, I. Glenn Cohen

Law Faculty Scholarship

[Excerpt] "The FDA’s regulation of drugs is frequently the subject of policy debate, with arguments falling into two camps. On the one hand, a libertarian view of patients and the health care system holds high the value of consumer choice. Patients should get all the information and the drugs they want; the FDA should do what it can to enforce some basic standards but should otherwise get out of the way. On the other hand, a paternalist view values the FDA’s role as an expert agency standing between patients and a set of potentially dangerous drugs and potentially unscrupulous or …

Effect Of Origanum Chemotypes On Broiler Intestinal Bacteria, Liliana Betancourt, Fernando Rodriguez, Vienvilay Phandanouvong, Claudia Ariza-Nieto, Michael Hume, David Nisbet, German Afanador-Tellez, Alexandra Martynova-Van Kley, Armen Nalian

Faculty Publications

Essential oils have been proposed as alternatives to antibiotic use in food animal production. This study evaluated 3 chemotypes of the Origanum genus, containing varying amounts of secondary metabolites carvacrol, thymol, and sabinene, in the broiler chicken diet. Aerial parts of Origanum vulgare L. (OL), O. vulgare L. ssp. hirtum (OH), and O. majorana (OM) were collected from a greenhouse located in the high altitude Sabana de Bogotá (Savanna of Bogotá) and O. vulgare L. ssp. hirtum (OG) produced and ground in Greece. Oregano essential oils (OEO) from these plants were obtained by steam distillation and analyzed by gas chromatography …

Development Of Loop-Mediated Isothermal Amplification Combined With Colorimetry Using Nanogold Probe For Chlamydia Trachomatis Detection, Ilada Choopara

Chulalongkorn University Theses and Dissertations (Chula ETD)

Chlamydia trachomatis จัดเป็นแบคทีเรียที่ก่อโรคติดต่อทางเพศสัมพันธ์ที่พบมากที่สุดทั่วโลกทั้งในเพศหญิงและชาย ถึงแม้ว่าโรคติดเชื้อนี้จะไม่ทําให้ผู้ติดเชื้อถึงตายและไม่แสดงอาการเริ่มต้นหรือแสดงเพียงเล็กน้อยนั้น การติดเชื้อนี้ในระยะยาวมีส่วนทําให้เป็นหมันหรือมีบุตรยาก และส่งเสริมการเกิดมะเร็งปากมดลูกจากไวรัสเอชพีวีได้ จึงเป็นที่น่าเสียดายที่การตรวจสุขภาพประจําปียังไม่มีการตรวจเชื้อนี้รวมอยู่ด้วย และยังไม่มีวิธีการตรวจเชื้อนี้ในคลินิกหรือสถานีอนามัยต่างๆ ที่การตรวจเชื้อนี้มีความสําคัญ ปัจจุบันวิธีมาตรฐานที่ใช้ตรวจคือการวิเคราะห์อาการโดยแพทย์ หรือการวิเคราะห์ลําดับเบสของยีน ompA (วิธีนี้ประกอบด้วยขั้นตอนการสกัดดีเอ็นเอด้วยชุดสกัดสําเร็จรูป การทําปฏิกิริยาลูกโซ่โพลีเมอเรส การรันเจลอิเล็กโตรโฟริซิส และการหาลําดับเบส) ดังนั้น งานวิจัยนี้จึงได้พัฒนาวิธีที่ไม่ซับซ้อนและประหยัดค่าใช้จ่ายขึ้นจากเทคนิคลูปเมดิเอเตดไอโซเทอร์มอลแอมพลิฟิเคชัน (LAMP) ร่วมกับการวัดสีโดยใช้โพรบติดอนุภาคทองระดับนาโน (AuNP) พบว่าวิธีที่พัฒนาขึ้นให้ผลความจําเพาะและความไวเทียบเท่าวิธีมาตรฐาน และใช้เวลาเพียง 24.5 นาที ขณะที่วิธีมาตรฐานใช้เวลามากกว่า 24 ชั่วโมง เมื่อรวมกับวิธีสกัดดีเอ็นเออย่างหยาบที่เหมาะสมกับชนิดของตัวอย่างพบว่าใช้เวลารวมทั้งสิ้นเพียง 29.5 นาที และมีค่าใช้จ่ายเพียง $3.6 ขณะที่วิธีมาตรฐานราคา $13.8 นอกจากนี้เมื่อทําการสุ่มตรวจในกลุ่มคนไข้และกลุ่มคนปกติ จํานวน 130 คน พบว่าวิธีที่พัฒนาขึ้นให้ผลตรงกับวิธีมาตรฐาน

Evidence That The C‑Terminal Domain Of A Type B Puta Protein Contributes To Aldehyde Dehydrogenase Activity And Substrate Channeling, Min Luo, Shelbi Christgen, Nikhilesh Sanyal, Benjamin W. Arentson, Donald F. Becker, John J. Tanner

Department of Biochemistry: Faculty Publications

Proline utilization A (PutA) is a bifunctional enzyme that catalyzes the oxidation of proline to glutamate. Structures of type A PutAs have revealed the catalytic core consisting of proline dehydrogenase (PRODH) and Δ1- pyrroline-5-carboxylate dehydrogenase (P5CDH) modules connected by a substrate-channeling tunnel. Type B PutAs also have a C-terminal domain of unknown function (CTDUF) that is absent in type A PutAs. Small-angle X-ray scattering (SAXS), mutagenesis, and kinetics are used to determine the contributions of this domain to PutA structure and function. The 1127-residue Rhodobacter capsulatus PutA (RcPutA) is used as a representative CTDUF-containing type B PutA. The reaction progress …

Evidence For Hysteretic Substrate Channeling In The Proline Dehydrogenaseand ∆1-Pyrroline-5-Carboxylate Dehydrogenase Coupled Reaction Of Proline Utilizationa(Puta), Michael A Moxley, Nikhilesh Sanyal, Navasona Krishnan, John J. Tanner, Donald F. Becker

Department of Biochemistry: Faculty Publications

Background: PutA from Escherichia coli is a bifunctional enzyme and transcriptional repressor in proline catabolism.

Results: Steady-state and transient kinetic data revealed a mechanism in which the two enzymatic reactions are coupled by an activation step.

Conclusion: Substrate channeling in PutA exhibits hysteretic behavior.

Significance: This is the first kinetic model of bi-enzyme activity in PutA and reveals a novel mechanism of channeling activation.

New Clox Systems For Rapid And Efficient Gene Disruption In Candida Albicans, Shahida Shahana, Delma S. Childers, Elizabeth R. Ballou, Iryna Bohovych, Frank C. Odds, Neil A.R. Gow, Alistair J.P. Brown

Department of Biochemistry: Faculty Publications

Precise genome modification is essential for the molecular dissection of Candida albicans, and is yielding invaluable information about the roles of specific gene functions in this major fungal pathogen of humans. C. albicans is naturally diploid, unable to undergo meiosis, and utilizes a non-canonical genetic code. Hence, specialized tools have had to be developed for gene disruption in C. albicans that permit the deletion of both target alleles, and in some cases, the recycling of the Candida-specific selectable markers. Previously, we developed a tool based on the Cre recombinase, which recycles markers in C. albicans with 90–100% efficiency via …

Kinetic And Structural Characterization Of Tunnel-Perturbing Mutants In Bradyrhizobium Japonicum Proline Utilization A, Benjamin W. Arentson, Min Luo, Travis A. Pemberton, John J. Tanner, Donald F. Becker

Department of Biochemistry: Faculty Publications

Proline utilization A from Bradyrhizobium japonicum (BjPutA) is a bifunctional flavoenzyme that catalyzes the oxidation of proline to glutamate using fused proline dehydrogenase (PRODH) and Δ¹-pyrroline-5-carboxylate dehydrogenase (P5CDH) domains. Recent crystal structures and kinetic data suggest an intramolecular channel connects the two active sites, promoting substrate channeling of the intermediate Δ¹-pyrroline-5-carboxylate/glutamate-γ-semialdehyde (P5C/GSA). In this work, the structure of the channel was explored by inserting large side chain residues at four positions along the channel in BjPutA. Kinetic analysis of the different mutants revealed replacement of D779 with Tyr (D779Y) or Trp (D779W) significantly decreased the overall …

Hepatoprotective Role Of Sestrin2 Against Chronic Er Stress, Hwan-Woo Park, Haeli Park, Seung-Hyun Ro, Insook Jang, Ian A. Semple, David N. Kim, Myungjin Kim, Myeongjin Nam, Deqiang Zhang, Lei Yin, Jun Hee Lee

Department of Biochemistry: Faculty Publications

Upon prolonged endoplasmic reticulum (ER) stress, cells attenuate protein translation to prevent accumulation of unfolded proteins. Here we show that Sestrin2 is critical for this process. Sestrin2 expression is induced by an ER stress-activated transcription factor CCAATenhancer- binding protein beta (c/EBPβ). Once induced, Sestrin2 halts protein synthesis by inhibiting mammalian target of rapamycin complex 1 (mTORC1). As Sestrin2-deficient cells continue to translate a large amount of proteins during ER stress, they are highly susceptible to ER stress-associated cell death. Accordingly, dietary or genetically induced obesity, which does not lead to any pathological indication other than simple fat accumulation in the …

Mechanisms Underlying The Exquisite Sensitivity Of Candida Albicans To Combinatorial Cationic And Oxidative Stress That Enhances The Potent Fungicidal Activity Of Phagocytes, Despoina Kaloriti, Mette Jacobsen, Zhikang Yin, Miranda Patterson, Anna Tillmann, Deborah A. Smith, Emily Cook, Tao You, Melissa J. Grimm, Iryna Bohovych, Celso Grebogi, Brahm H. Segal, Neil A.R. Gow, Ken Haynes, Janet Quinn, Alistair J.P. Brown

Department of Biochemistry: Faculty Publications

Immune cells exploit reactive oxygen species (ROS) and cationic fluxes to kill microbial pathogens, such as the fungus

Candida albicans. Yet, C. albicans is resistant to these stresses in vitro. Therefore, what accounts for the potent antifungal activity

of neutrophils? We show that simultaneous exposure to oxidative and cationic stresses is much more potent than the individual

stresses themselves and that this combinatorial stress kills C. albicans synergistically in vitro.We also show that the high

fungicidal activity of human neutrophils is dependent on the combinatorial effects of the oxidative burst and cationic fluxes, as

their pharmacological attenuation with …

Oxidative Stress, Redox Signaling, And Autophagy: Cell Death Versus Survival, Juliana Navarro-Yepes, Michaela Burns, Anandhan Annadurai, Oleh Khalimonchuk, Luz Maria Del Razo, Betzabet Quintanilla-Vega, Aglaia Pappa, Mihalis I. Panayiotidis, Rodrigo Franco

Department of Biochemistry: Faculty Publications

Significance: The molecular machinery regulating autophagy has started becoming elucidated, and a number of studies have undertaken the task to determine the role of autophagy in cell fate determination within the context of human disease progression. Oxidative stress and redox signaling are also largely involved in the etiology of human diseases, where both survival and cell death signaling cascades have been reported to be modulated by reactive oxygen species (ROS) and reactive nitrogen species (RNS). Recent Advances: To date, there is a good understanding of the signaling events regulating autophagy, as well as the signaling processes by which alterations in …

Evaluation Of Three Herbicide Resistance Genes For Use In Genetic Transformations And For Potential Crop Protection In Algae Production, Andrew J. Bruggeman, Daniel Kuehler, Donald P. Weeks

Department of Biochemistry: Faculty Publications

Genes conferring resistance to the herbicides glyphosate, oxyfluorfen and norflurazon were developed and tested for use as dominant selectable markers in genetic transformation of Chlamydomonas reinhardtii and as potential tools for the protection of commercial-scale algal production facilities against contamination by organisms sensitive to these broad-spectrum herbicides. A synthetic glyphosate acetyltransferase (GAT) gene, when fitted with a strong Chlamydomonas promoter, conferred a 2.79-fold increase in tolerance to the EPSPS inhibitor, glyphosate, in transgenic cells compared with progenitor WT cells. A mutant Chlamydomonas protoporphyrinogen oxidase (protox, PPO) gene previously shown to produce an enzyme insensitive to PPO-inhibiting herbicides, when genetically engineered, …

Agricultural Biotechnology-An Opportunity To Feed A World Of Ten Billion, Nina V. Fedoroff, Drew L. Kershen

Faculty Articles

No abstract provided.