Biotechnology Commons^™

Generating Pennycress (Thlaspi Arvense) Seed Triacylglycerols And Acetyl-Triacylglycerols Containing Medium-Chain Fatty Acids, Maliheh Esfahanian, Tara J. Nazarenus, Meghan M. Freund, Gary Mcintosh, Winthrop B. Phippen, Mary E. Phippen, Timothy P. Durrett, Edgar B. Cahoon, John C. Sedbrook

Department of Biochemistry: Faculty Publications

Thlaspi arvense L. (pennycress) is a cold-tolerant Brassicaceae that produces large amounts of seeds rich in triacylglycerols and protein, making it an attractive target for domestication into an offseason oilseed cash cover crop. Pennycress is easily genetically transformed, enabling synthetic biology approaches to tailor oil properties for specific biofuel and industrial applications. To test the feasibility in pennycress of producing TAGs and acetyl-TAGs rich in medium-chain fatty acids (MCFAs; C6–C14) for industrial, biojet fuel and improved biodiesel applications, we generated transgenic lines with seed-specific expression of unique acyltransferase (LPAT and diacylglycerol acyltransferase) genes and thioesterase (FatB) genes isolated from Cuphea …

Genetic Engineering Of Lesquerella With Increased Ricinoleic Acid Content In Seed Oil, Grace Q. Chen, Kumiko Johnson, Tara J. Nazarenus, Grisel Ponciano, Eva Morales, Edgar B. Cahoon

Department of Biochemistry: Faculty Publications

Seeds of castor (Ricinus communis) are enriched in oil with high levels of the industrially valuable fatty acid ricinoleic acid (18:1OH), but production of this plant is limited because of the cooccurrence of the ricin toxin in its seeds. Lesquerella (Physaria fendleri) is being developed as an alternative industrial oilseed because its seeds accumulate lesquerolic acid (20:1OH), an elongated form of 18:1OH in seed oil which lacks toxins. Synthesis of 20:1OH is through elongation of 18:1OH by a lesquerella elongase, PfKCS18. Oleic acid (18:1) is the substrate for 18:1OH synthesis, but it is also used by fatty acid desaturase 2 …

Long Non-Coding Rna Meg3 Deficiency Impairs Glucose Homeostasis And Insulin Signaling By Inducing Cellular Senescence Of Hepatic Endothelium In Obesity, Xiao Cheng, Mohamed Sham Shihabudeen Haider Ali, Matthew Moran, Martonio Ponte Viana, Sarah L. Schlichte, Matthew C. Zimmerman, Oleh Khalimonchuk, Mark W. Feinberg, Xinghui Sun

Department of Biochemistry: Faculty Publications

Obesity-induced insulin resistance is a risk factor for diabetes and cardiovascular disease. However, the mechanisms underlying endothelial senescence in obesity, and how it impacts obesity-induced insulin resistance remain incompletely understood. In this study, transcriptome analysis revealed that the long non-coding RNA (lncRNA) Maternally expressed gene 3 (Meg3) is one of the top differentially expressed lncRNAs in the vascular endothelium in diet-induced obese mice. Meg3 knockdown induces cellular senescence of endothelial cells characterized by increased senescence-associated β–galactosidase activity, increased levels of endogenous superoxide, impaired mitochondrial structure and function, and impaired autophagy. Moreover, Meg3 knockdown causes cellular senescence of hepatic endothelium in …

Resistance To Aflatoxin Accumulation In Maize Mediated By Host-Induced Gene Silencing Of Aspergillus Flavus Alkaline Protease And O-Methyltransferase-A Genes, Olanike Omotola Omolehin

LSU Doctoral Dissertations

Aspergillus flavus is a soil-borne fungal pathogen that infects maize and produces aflatoxins. In the current study, portions of the alkaline protease (alk) and the O-methyl transferase (omtA) genes, which are key in A. flavus virulence and aflatoxin biosynthesis, respectively, were targeted for suppression through an RNAi (RNA interference) approach known as Host-Induced Gene Silencing (HIGS). Separate RNAi vectors were designed to carry regions of the alk and omtA gene fragments (Alk-RNAi and OmtA-RNAi) and introduced into B104 maize zygotic embryos. Eight and six transformation events were positive for the alk and omtA transgene, respectively. …

Suspicion Encoded: Women Of Color And Biometric Technology In The United States, Lilith A. Saylor

sprinkle: an undergraduate journal of feminist and queer studies

This paper explores the commodification of privacy through biometric technology in the United States. It examines the impact of this technology on poor women of color (WOC), arguing that poor WOC face intersectional discrimination based on the convergence of sex, race, and class in their identities. I highlight the unique and powerful intrusion of biometric technology into the lives of poor WOC, and argue that the connection between data and the physical body created through biometric data has formed an environment in which the state wields unrestricted control in all spheres over the privacy of poor WOC.

Root Nrt, Nir, Amt, Gs, Gogat And Gdh Expression Levels Reveal No Andaba Mediated Drought Tolerance In Brassica Juncea L., Seema Sahay, Luis Robledo-Arratia, Katarzyna Glowacka, Meetu Gupta

Department of Biochemistry: Faculty Publications

Little is known about the interactive effects of exogenous nitric oxide (NO) and abscisic acid (ABA) on nitrogen (N) metabolism and related changes at molecular and biochemical levels under drought stress. The present study highlights the independent and combined effect of NO and ABA (grouped as “nitrate agonists”) on expression profiles of representative key genes known to be involved in N-uptake and assimilation, together with proline metabolism, N–NO metabolism enzyme’s activity and nutrient content in polyethylene glycol (PEG) treated roots of Indian mustard (B. juncea cv. Varuna). Here we report that PEG mediated drought stress negatively inhibited growth performance, as …

Development Of A Dna Biosensor Using Crispr/Dcas9 On A Graphene Oxide Surface For Detecting Antibiotic Resistance Genes, Wendy Cecil

Mahurin Honors College Capstone Experience/Thesis Projects

Antibiotic resistance is a rapidly spreading global threat to human health. Previously in our lab, we developed a simple method for facile detection of antibiotic resistance genes (ARGs) in bacteria. This DNA detection system takes advantage of the CRISPR-Cas system (Clustered Regularly Interspaced Short Palindromic Repeat and CRISPRAssociated Protein). We designed CRISPR in complex with nuclease deactivated Cas9 (dCas9) on graphene oxide (GO) sheets conjugated with Cas9 antibodies. Single guide RNA (sgRNA) was customized to program dCas9 for target DNA binding. sgRNA was covalently labeled with fluorescein to provide fluorescence signal. Graphene oxide (GO) is utilized for its quenching properties …

The Role Of Ms-818 In Altering Age-Related Characteristics Of An In Vitro Model Of Senescence In Neural Stem Cells, Sandeep Sreerama

Honors Undergraduate Theses

Aging of the brain is the leading risk factor for neurodegenerative diseases and brain cancers and has deleterious effects on brain functions. It follows that attempts to reverse the aging process may be therapeutically valuable. Neural stem cells (NSC) have been shown to play a critical role in maintaining brain functions, and their number is severely decreased with age. The development of senescence-like characteristics and declining functions in NSCs have been proposed to be responsible for brain aging and tumorigenesis. MS-818 is a pyrrolopyrimidine that has been shown to increase the NSC population and reverse the decline of behavioral function …

Teaching Metabolism In Upper-Division Undergraduate Biochemistry Courses Using Online Computational Systems And Dynamical Models Improves Student Performance, Christine S. Booth, Changsoo Song, Michelle E. Howell, Achilles Rasquinha, Aleš Saska, Resa M. Helikar, Sharmin M. Sikich, Brain A. Couch, Karin Van Dijk, Rebecca Roston, Tomáš Helikar

Department of Biochemistry: Faculty Publications

Understanding metabolic function requires knowledge of the dynamics, interdependence, and regulation of metabolic networks. However, multiple professional societies have recognized that most undergraduate biochemistry students acquire only a surface-level understanding of metabolism. We hypothesized that guiding students through interactive computer simulations of metabolic systems would increase their ability to recognize how individual interactions between components affect the behavior of a system under different conditions. The computer simulations were designed with an interactive activity (i.e., module) that used the predict–observe–explain model of instruction to guide students through a process in which they iteratively predict outcomes, test their predictions, modify the interactions …

Long Non-Coding Rna Meg3 Deficiency Impairs Glucose Homeostasis And Insulin Signaling By Inducing Cellular Senescence Of Hepatic Endothelium In Obesity, Xiao Cheng, Mohamed Sham Shihabudeen Haider Ali, Matthew Moran, Martonio Ponte Viana, Sarah L. Schlichte, Matthew C. Zimmerman, Oleh Khalimonchuk, Mark W. Feinberg, Xinghui Sun

Department of Biochemistry: Faculty Publications

Obesity-induced insulin resistance is a risk factor for diabetes and cardiovascular disease. However, the mechanisms underlying endothelial senescence in obesity, and how it impacts obesity-induced insulin resistance remain incompletely understood. In this study, transcriptome analysis revealed that the long non-coding RNA (lncRNA) Maternally expressed gene 3 (Meg3) is one of the top differentially expressed lncRNAs in the vascular endothelium in diet-induced obese mice. Meg3 knockdown induces cellular senescence of endothelial cells characterized by increased senescence-associated β–galactosidase activity, increased levels of endogenous superoxide, impaired mitochondrial structure and function, and impaired autophagy. Moreover, Meg3 knockdown causes cellular senescence of hepatic endothelium in …

Integrative Network Analyses Of Transcriptomics Data Reveal Potential Drug Targets For Acute Radiation Syndrome, Robert Moore, Bhanwar Lal Puniya, Robert Powers, Chittibabu Guda, Kenneth W. Bayles, David Berkowitz, Tomas Helikar

Department of Biochemistry: Faculty Publications

Recent political unrest has highlighted the importance of understanding the short- and long-term effects of gamma-radiation exposure on human health and survivability. In this regard, effective treatment for acute radiation syndrome (ARS) is a necessity in cases of nuclear disasters. Here, we propose 20 therapeutic targets for ARS identified using a systematic approach that integrates gene co-expression networks obtained under radiation treatment in humans and mice, drug databases, disease-gene association, radiation-induced differential gene expression, and literature mining. By selecting gene targets with existing drugs, we identified potential candidates for drug repurposing. Eight of these genes (BRD4, NFKBIA, CDKN1A, TFPI, MMP9, …

Root Nrt, Nir, Amt, Gs, Gogat And Gdh Expression Levels Reveal No Andaba Mediated Drought Tolerance In Brassica Juncea L., Seema Sahay, Luis Robledo-Arratia, Katarzyna Glowacka, Meetu Gupta

Department of Biochemistry: Faculty Publications

Little is known about the interactive effects of exogenous nitric oxide (NO) and abscisic acid (ABA) on nitrogen (N) metabolism and related changes at molecular and biochemical levels under drought stress. The present study highlights the independent and combined effect of NO and ABA (grouped as “nitrate agonists”) on expression profiles of representative key genes known to be involved in N-uptake and assimilation, together with proline metabolism, N–NO metabolism enzyme’s activity and nutrient content in polyethylene glycol (PEG) treated roots of Indian mustard (B. juncea cv. Varuna). Here we report that PEG mediated drought stress negatively inhibited growth performance, as …

Lysosomal Slc46a3 Modulates Hepatic Cytosolic Copper Homeostasis, Jung-Hwan Kim, Tsutomu Matsubara, Jaekwom Lee, Cristina Fenollar-Ferrer, Kyungreem Han, Donghwan Kim, Shang Jia, Christopher J. Chang, Heejung Yang, Tomokazu Nagano, Kristopher W. Krausz, Sun Hee Yim, Frank J. Gonzalez

Department of Biochemistry: Faculty Publications

The environmental contaminant 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD) causes hepatic toxicity associated with prominent lipid accumulation in humans. Here, the authors report that the lysosomal copper transporter SLC46A3 is induced by TCDD and underlies the hepatic lipid accumulation in mice, potentially via effects on mitochondrial function. SLC46A3 was localized to the lysosome where it modulated intracellular copper levels. Forced expression of hepatic SLC46A3 resulted in decreased mitochondrial membrane potential and abnormal mitochondria morphology consistent with lower copper levels. SLC46A3 expression increased hepatic lipid accumulation similar to the known effects of TCDD exposure in mice and humans. The TCDD-induced hepatic triglyceride accumulation was significantly …

Covid19 Disease Map, A Computational Knowledge Repository Of Virus–Host Interaction Mechanisms, Marek Ostaszewski, Tomáš Helikar, Bhanwar Lal Puniya, A Host Of Co-Authors, Covid-19 Disease Map Community

Department of Biochemistry: Faculty Publications

We need to effectively combine the knowledge from surging literature with complex datasets to propose mechanistic models of SARS-CoV-2 infection, improving data interpretation and predicting key targets of intervention. Here, we describe a large-scale community effort to build an open access, interoperable and computable repository of COVID-19 molecular mechanisms. The COVID-19 Disease Map (C19DMap) is a graphical, interactive representation of disease-relevant molecular mechanisms linking many knowledge sources. Notably, it is a computational resource for graph-based analyses and disease modelling. To this end, we established a framework of tools, platforms and guidelines necessary for a multifaceted community of biocurators, domain experts, …

Association Of The Malate Dehydrogenase‑Citrate Synthase Metabolon Is Modulated By Intermediates Of The Krebs Tricarboxylic Acid Cycle, Joy Omini, Izabela Wojciechowska, Aleksandra Skirycz, Hideaki Moriyama, Toshihiro Obata

Department of Biochemistry: Faculty Publications

Mitochondrial malate dehydrogenase (MDH)-citrate synthase (CS) multi-enzyme complex is a part of the Krebs tricarboxylic acid (TCA) cycle ‘metabolon’ which is enzyme machinery catalyzing sequential reactions without diffusion of reaction intermediates into a bulk matrix. This complex is assumed to be a dynamic structure involved in the regulation of the cycle by enhancing metabolic flux. Microscale Thermophoresis analysis of the porcine heart MDH-CS complex revealed that substrates of the MDH and CS reactions, NAD+ and acetyl-CoA, enhance complex association while products of the reactions, NADH and citrate, weaken the affinity of the complex. Oxaloacetate enhanced the interaction only when it …

Lipid Droplet Membrane Proteome Remodeling Parallels Ethanol-Induced Hepatic Steatosis And Its Resolution, Carol A. Casey, Terrence M. Donohue Jr., Jacy L. Kubik, Vikas Kumar, Michael J. Naldrett, Nicholas T. Woods, Cole P. Frisbie, Mark A. Mcniven, Paul G. Thomes

Nebraska Center for Biotechnology: Faculty and Staff Publications

Abstract Lipid droplets (LDs) are composed of neutral lipids enclosed in a phospholipid monolayer, which harbors membrane-associated proteins that regulate LD functions. Despite the crucial role of LDs in lipid metabolism, remodeling of LD protein composition in disease contexts, such as steatosis, remains poorly understood. We hypothesized that chronic ethanol consumption, subsequent abstinence from ethanol, or fasting differentially affects the LD membrane proteome content and that these changes influence how LDs interact with other intracellular organelles. Here, male Wistar rats were pair-fed liquid control or ethanol diets for 6 weeks, and then, randomly chosen animals from both groups were either …

Visualizing Barrier Dysfunction Using Clarity, Rachel Robbe

Lewis Honors College Thesis Collection

Epilepsy and Alzheimer’s disease are two prevalent neurological disorders. The blood-brain barrier (BBB) is a factor in why there are no long-term treatments for these diseases. The transporters in the neurovascular unit (NVU), which makes up the BBB, are highly specialized, so most drugs cannot pass the barrier to treat these diseases. It is unknown why drug uptake in the BBB varies in a diseased and wild-type brain, but one theory is that there are spatial differences in the NVU. One method for examining the NVU is the CLARITY tissue clearing technique which is unique in its capability to clear …

การวิเคราะห์ปัจจัยสำหรับการเกิดคราบบนแม่พิมพ์ระหว่างวัลคาไนเซชันยางด้วยการออกแบบการทดลองเชิงแฟกทอเรียล, สิรีธร อุตมโชติ

Chulalongkorn University Theses and Dissertations (Chula ETD)

ในอุตสาหกรรมการผลิตชิ้นส่วนยางพบปัญหาจากการเกิดชิ้นงานที่มีจุดบกพร่อง เช่น พอง ฉีกขาด แผล และคราบ โดยชิ้นงานที่เป็นคราบ คือ ชิ้นงานที่ผิวที่ไม่เรียบและต้องหยุดการผลิตเพื่อนำแม่พิมพ์ไปทำความสะอาด ทำให้ต้นทุนการผลิตเพิ่มขึ้น สาเหตุของการเกิดคราบถูกนำมาวิเคราะห์ด้วยวิธีการออกแบบการทดลองเชิงแฟกทอเรียลเพื่อหาความสำคัญทางสถิติของแต่ละตัวแปร โดยตัวแปรที่ศึกษาได้แก่ เวลาที่ใช้ในการขึ้นรูป ปริมาณซิงค์ออกไซด์ ปริมาณแคลเซียมคาร์บอเนต และอัตราส่วนโดยน้ำหนักของน้ำยาถอดแบบต่อน้ำปราศจากไอออน จากผลการทดลองเชิงแฟกทอเรียล พบว่าเวลาที่ใช้ในการขึ้นรูปและอัตราส่วนโดยน้ำหนักของน้ำยาถอดแบบต่อน้ำปราศจากไอออน คือ ตัวแปรสำคัญที่ทำให้ชิ้นงานเกิดคราบ เนื่องจากการใช้เวลาในการขึ้นรูปที่นานขึ้น ทำให้ยางเกิดระดับการวัลคาไนเซชันเพิ่มขึ้น ซึ่งทำให้แรงยึดติดระหว่างชิ้นงานกับแม่พิมพ์ลดลง ส่งผลให้ทำให้มีคราบที่เกิดขึ้นลดลง และเมื่ออัตราส่วนโดยน้ำหนักของน้ำยาถอดแบบต่อน้ำปราศจากไอออนที่ 1:100 พบว่ามีชิ้นงานที่เป็นคราบลดลง เนื่องจากน้ำยาถอดแบบทำให้ผิวของแม่พิมพ์มีความลื่น จึงทำให้ไม่มีสิ่งตกค้างหลงเหลืออยู่ที่สามารถนำไปสู่การเกิดคราบบนผิวของแม่พิมพ์ได้ โดยแนวโน้มของการเกิดคราบต่อการปรับเปลี่ยนเวลาในการขึ้นรูปและอัตราส่วนของน้ำยาถอดแบบได้มาจากการทดลองเชิงเดี่ยว พบว่าเมื่อเวลาการขึ้นรูปมากขึ้นจาก 350 ไปเป็น 450 วินาที และอัตราส่วนโดนน้ำหนักของน้ำยาถอดแบบต่อน้ำปราศจากไอออนจาก 1:450 ไปเป็น 1:250 ทำให้มีชิ้นงานที่เป็นคราบลดลง

Use Of Lymesim 2.0 To Assess The Potential For Single And Integrated Management Methods To Control Blacklegged Ticks (Ixodes Scapularis; Acari: Ixodidae) And Transmission Of Lyme Disease Spirochetes, Shravani Chitineni, Elizabeth R. Gleim, Holly D. Gaff

Undergraduate Honors Theses

Annual Lyme disease cases continue to rise in the U.S. making it the most reported vector-borne illness in the country. The pathogen (Borrelia burgdorferi) and primary vector (Ixodes scapularis; blacklegged tick) dynamics of Lyme disease are complicated by the multitude of vertebrate hosts and varying environmental factors, making models an ideal tool for exploring disease dynamics in a time- and cost-effective way. In the current study, LYMESIM 2.0, a mechanistic model, was used to explore the effectiveness of three commonly used tick control methods: habitat-targeted acaricide (spraying), rodent-targeted acaricide (bait boxes), and white-tailed deer targeted acaricide (4-poster …

Late-Stage Product Development And Approvals By Biotechnology Companies After Initial Public Offering, 1997-2016, Laura M. M. Mcnamee, Ekaterina Galkina Cleary, Sunyi Zhang, Usama Salim, Fred D. Ledley

Natural & Applied Sciences Faculty Publications

Purpose: This work describes the late-stage product portfolios of the biotechnology companies that completed initial public offerings (IPOs) from 1997 to 2016. We asked whether these emerging companies continue to develop innovative, biologic products and produce the innovation promised by the early biotechnology industry.

Methods: We identified therapeutic products that reached Phase III development from 1997 to 2016, the characteristics of the products, the dates of the initiation of Phase III and product approval, proxy indicators of the innovativeness of each product, and the contribution of each biotechnology company. Companies were characterized by IPO window and clinical status of the …