Biotechnology Commons^™

Ligand Binding And Signaling Of Hare/Stabilin-2, Edward N. Harris, Fatima Cabral

Department of Biochemistry: Faculty Publications

The Stabilin receptors are a two-member family in the type H class of scavenger receptors. These dynamic receptors bind and internalize multiple ligands from the cell surface for the purpose of clearing extracellular material including some synthetic drugs and for sensing the external environment of the cell. Stabilin-1 was the first receptor to be cloned, though the biological activity of Hyaluronic Acid Receptor for Endocytosis (HARE)/Stabilin-2 was observed about 10 years prior to the cloning of Stabilin-1. Stabilin-1 has a more diverse expression profile among the tissues than HARE/Stabilin-2. This review will focus on HARE/Stabilin-2 and its interactions with hyaluronan, …

Genome‐Wide Association And Genomic Prediction For Biomass Yield In A Genetically Diverse Miscanthus Sinensis Germplasm Panel Phenotyped At Five Locations In Asia And North America, Lindsay V. Clark, Maria S. Dwiyanti, Kossonou G. Anzoua, Joe E. Brummer, Bimal Kumar Ghimire, Katarzyna Glowacka, Megan Hall, Kweon Heo, Xiaoli Jin, Alexander E. Lipka, Junhua Peng, Toshihiko Yamada, Ji Hye Yoo, Chang Yeon Yu, Hua Zhao, Stephen P. Long, Erik J. Sacks

Department of Biochemistry: Faculty Publications

To improve the efficiency of breeding of Miscanthus for biomass yield, there is a need to develop genomics‐assisted selection for this long‐lived perennial crop by relating genotype to phenotype and breeding value across a broad range of environments. We present the first genome‐wide association (GWA) and genomic prediction study of Miscanthus that utilizes multilocation phenotypic data. A panel of 568 Miscanthus sinensis accessions was genotyped with 46,177 single nucleotide polymorphisms (SNPs) and evaluated at one subtropical and five temperate locations over 3 years for biomass yield and 14 yield‐component traits. GWA and genomic prediction were performed separately for different years …

Downregulation Of A Cyp74 Rubber Particle Protein Increases Natural Rubber Production In Parthenium Argentatum, Dante F. Placido, Niu Dong, Chen Dong, Von Mark V. Cruz, David A. Dierig, Rebecca E. Cahoon, Byung-Guk Kang, Trinh Huynh, Maureen Whalen, Grisel Ponciano, Colleen Mcmahan

Department of Biochemistry: Faculty Publications

We report functional genomics studies of a CYP74 rubber particle protein from Parthenium argentatum, commonly called guayule. Previously identified as an allene oxide synthase (AOS), this CYP74 constitutes the most abundant protein found in guayule rubber particles. Transgenic guayule lines with AOS gene expression down-regulated by RNAi (AOSi) exhibited strong phenotypes that included agricultural traits conducive to enhancing rubber yield. AOSi lines had higher leaf and stem biomass, thicker stembark tissues, increased stem branching and improved net photosynthetic rate. Importantly, the rubber content was significantly increased in AOSi lines compared to the wild-type (WT), vector control and …

การผลิตไบโอดีเซลจากกรดไขมันอิสระของน้ำมันปาล์มโดยปฏิกิริยาสองขั้นตอนโดยใช้ตัวเร่งปฏิกิริยาชีวภาพทั้งเซลล์, ฐนิสา หวั่งประดิษฐ์

Chulalongkorn University Theses and Dissertations (Chula ETD)

การผลิตไบโอดีเซลสามารถผลิตได้จากกรดไขมันอิสระของน้ำมันปาล์มโดยปฏิกิริยาสองขั้นตอนโดยใช้ตัวเร่งปฏิกิริยาชีวภาพทั้งเซลล์ ในการศึกษานี้ได้ใช้ยีสต์ Candida rugosa เป็นตัวเร่งปฏิกิริยาชีวภาพทั้งเซลล์ เนื่องจากเชื้อยีสต์ดังกล่าวมีความสามารถในการไฮโดรไลซ์น้ำมันปาล์มไปเป็นกรดไขมันอิสระ โดยพบว่าได้กรดไขมันอิสระสูงสุดเท่ากับ 82.61 เปอร์เซ็นต์ หลังจากทำการเพาะเลี้ยงเป็นเวลา 5 วัน จากนั้นกรดไขมันอิสระที่ผลิตโดย C. rugosa จะถูกนำไปใช้เป็นตัวเหนี่ยวนำในการผลิตเอนไซม์ลิเพสจากเชื้อ Aureobasidium pullulans var. melanogenum SRY 14-3 ซึ่งหลังจากการหาภาวะที่เหมาะสมพบว่าสามารถผลิตเอนไซม์ลิเพสที่มีค่าแอกทิวิตีและค่าแอกทิวิตีจำเพาะของเอนไซม์สูงสุด คือ 1.18 ยูนิตต่อมิลลิลิตร และ 17.28 ยูนิตต่อมิลลิกรัมโปรตีน ตามลำดับ โดยใช้ความเข้มข้นของกรดไขมันอิสระ 3 เปอร์เซ็นต์ ที่อุณหภูมิ 30 องศาเซลเซียส เป็นเวลา 120 ชั่วโมง การศึกษาการผลิตไบโอดีเซลจากปฏิกิริยาเอสเทอริฟิเคชันที่เร่งปฏิกิริยาด้วย A. pullulans var. melanogenum SRY 14-3 พบว่าสามารถผลิตไบโอดีเซลได้สูงสุดเท่ากับ 21.39 เปอร์เซ็นต์ เมื่อใช้อัตราส่วนโดยโมลของกรดไขมันอิสระต่อเมทานอลเท่ากับ 1:1 ที่อุณหภูมิ 30 องศาเซลเซียส เป็นเวลา 72 ชั่วโมง สำหรับการศึกษาตัวเร่งปฏิกิริยาชีวภาพทั้งเซลล์ด้วยการใช้เทคนิคการแสดงออกที่ผิวเซลล์ยีสต์ เอนไซม์ลิเพส 1 และเอนไซม์ลิเพส 3 จาก C. rugosa (CRL1 และ CRL3) และเอนไซม์ลิเพส จาก A. pullulans var. melanogenum SRY 14-3 (AML) ได้ถูกนำมาเชื่อมต่อกับโปรตีนฐาน PpPIR1 แล้วทำการโคลนเข้าสู่ยีสต์เจ้าบ้าน Pichia pastoris KM 71 ได้เป็นยีสต์ Pp-CRL1, Pp-CRL3 และ Pp-AML ตามลำดับ จากการศึกษาพบว่ายีสต์สายพันธุ์รีคอมบิแนนท์ Pp-CRL1 และ Pp-CRL3 สามารถผลิตเอนไซม์ลิเพสที่ผิวเซลล์สูงสุดเท่ากับ 1,499.9 มิลลิยูนิตต่อOD600 และ 181.96 มิลลิยูนิตต่อOD600 …

A 3d Bioprinter Platform For Mechanistic Analysis Of Tumoroids And Chimeric Mammary Organoids, John A. Reid, Xavier-Lewis Palmer, Peter A. Mollica, Nicole Northam, Patrick C. Sachs, Robert D. Bruno

School of Medical Diagnostics & Translational Sciences Publications

The normal mammary microenvironment can suppress tumorigenesis and redirect cancer cells to adopt a normal mammary epithelial cell fate in vivo. Understanding of this phenomenon offers great promise for novel treatment and detection strategies in cancer, but current model systems make mechanistic insights into the process difficult. We have recently described a low-cost bioprinting platform designed to be accessible for basic cell biology laboratories. Here we report the use of this system for the study of tumorigenesis and microenvironmental redirection of breast cancer cells. We show our bioprinter significantly increases tumoroid formation in 3D collagen gels and allows for precise …

Malate Dehydrogenase Allele Frequencies In Honeybee Populations And Its Correlation With Temperature, Thitipan Meemongkolkiat

Chulalongkorn University Theses and Dissertations (Chula ETD)

Temperature is one of the major factors that affect on the variation in honeybee in term of DNA and protein variants. The aim of this study is to investigate whether the three structural variants of the cytosolic malate dehydrogenase (cMDH) protein of the honeybee can explain clinal variation in allele frequencies. To test the hypothesis that there is a molecular basis to the temperature-dependent fitness of cMDH variation, the enzymic activity was determined across a range of temperatures in vitro. The F and S alleles have higher activity and are less sensitive to high temperatures than the M allele. Next, …

Therapeutic Delivery Technology And Its Economic Impact, Paul E. Savas Jr.

Senior Honors Theses

Therapeutic delivery technology is a current area of high interest in both university and industrial settings. These technologies are being developed in order to deliver therapeutic agents, such as genes, proteins, and drugs, to patients more efficiently. Nanoscale delivery vehicles have proven to be useful for these applications; these vehicles may either be naturally produced or chemically synthesized. The physical properties of these nanomaterials must be characterized correctly using instrumentation that evaluates their size, morphology, and potential for agglomeration. These technologies represent a high-growth economic area that fosters entrepreneurship and innovation. Because of this innovative spirit, research and economic interest …

Unraveling The Role Of Phenylethanolamine N-Methyltransferase (Pnmt+) Cells In-Vivo, Sanjana Manja

Electronic Theses and Dissertations

Phenylethanolamine N-methyltransferase (Pnmt) is the enzyme that N-methylates norepinephrine to produce the stress hormone/neurotransmitter, epinephrine, which is abundantly expressed in adrenal glands. Developmental studies have also identified Pnmt expression in the embryonic heart and several areas of the brain, including brainstem, cerebellum, and hypothalamus. Thus, we hypothesize that selective ablation of Pnmt+ cells will have detrimental effects on cardiovascular, neuromuscular, and metabolic processes. To uncover the importance of Pnmt+ cells in vivo, we generated a novel Diphtheria Toxin A (DTA) suicide model (Pnmt+/Cre; R26+/DTA) to selectively ablate Pnmt-expressing (Pnmt+) cells in mice. Appearing normal at birth, Pnmt-Cre/DTA mice began to …

Bone Morphogenetic Protein 7 Inhibits Pyroptotic Cell Death In Vascular Smooth Muscle Cells Of Atherosclerotic Apolipoprotein E -/- Mice, Kaley Garner

Electronic Theses and Dissertations

Atherosclerosis (ATH) is an inflammation-mediated disease in which cell death underlies the formation of lesions along the intima layer of vascular walls resulting in vessel narrowing, decreased blood flow, and increased risk of lesion rupture leading to myocardial infarction and stroke. The current study was undertaken to investigate whether inflammation in ATH can induce pyroptosis in vascular smooth muscle cells (SMC's). We therefore hypothesized that pyroptosis occurs and is inhibited by bone morphogenetic protein 7 (BMP7). We examined SMC pyroptosis at acute (D5) and midstage (D28) following disturbed flow-induced hemodynamic injury to the vascular wall using our partial left carotid …

Toward An Enzyme-Coupled, Bioorthogonal Platform For Methyltransferases: Probing The Specificity Of Methionine Adenosyltransferases, Tyler D. Huber

Theses and Dissertations--Pharmacy

Methyl group transfer from S-adenosyl-l-methionine (AdoMet) to various substrates including DNA, proteins, and natural products (NPs), is accomplished by methyltransferases (MTs). Analogs of AdoMet, bearing an alternative S-alkyl group can be exploited, in the context of an array of wild-type MT-catalyzed reactions, to differentially alkylate DNA, proteins, and NPs. This technology provides a means to elucidate MT targets by the MT-mediated installation of chemoselective handles from AdoMet analogs to biologically relevant molecules and affords researchers a fresh route to diversify NP scaffolds by permitting the differential alkylation of chemical sites vulnerable to NP MTs that are unreactive to …

Mechanisms Of Microglia Mediated Apolipoprotien E Neurotoxicity, Pardeep Singh

Dissertations and Theses

No abstract provided.

Capital Efficiency For Development Stage Biotech-Based Firms: An Ipo Perspective, Mark J. Ahn, Amir Shaygan

Engineering and Technology Management Faculty Publications and Presentations

Access to multiple tranches of capital is critical for predominantly no revenue development stage biotech firms. While financing needs are monotonically increasing over multiple years in the product development approval cycle, the market for high risk, milestone driven biotech investment is significantly more volatile than the financial markets as a whole. In this paper, we analyzed the role and relative importance of global biotech IPOs, as well as other sources of capital such as strategic alliances, for research and development funding. We also explored and assessed the degree of mismatch between the access to capital, operational efficiencies, and how firms …

Preconditioning Of The Tumor Microenvironment By Means Of Low Dose Chemotherapies For An Effective Immunotherapy Of Breast Cancer, Hussein F. Aqbi

Theses and Dissertations

Breast cancer mortality is mainly due to distant recurrence of the disease arising from dormant tumor cells established by cancer therapies. Patients who initially respond to cancer therapies often succumb to distant recurrence of the disease. It is not clear why people with the same type of breast cancer respond to treatments differently; some escape from dormancy and relapse earlier than others. In addition, some tumor clones respond to immunotherapy while others do not. We investigated how autophagy plays a role in accelerating or delaying recurrence of neu overexpressing mouse mammary carcinoma (MMC) following adriamycin (ADR) treatment, and in affecting …

Cyclophilin A As A Molecular Switch Regulating Prolactin Receptor Mediated Signaling, Mammary Tumorigenesis And Metastasis, Shawn Hakim

Theses and Dissertations

Prolactin (PRL) and its receptor (PRLr) have been implicated in the development and progression of human breast cancer. PRL activates its receptor and induces activation of proximal Janus kinase 2 (Jak2) for signal transduction. Here, we sought to determine the role of PRLr-associated peptidyl-prolyl isomerase, cyclophilin A (CypA), in modulating structure/function relationships of the PRLr. It was demonstrated that CypA mediated PRL-induced conformational change of the CFP- and -YFP tagged forms of the PRLr cytoplasmic-tail, whereas CypA inhibition by NIM811 (N-methyl-4-isoleucine cyclosporin) or knockdown blocked the conformational change of the PRLr assessed by Fluorescence Resonance Energy Transfer (FRET) signal or …

Detection Of Sickle Cell Disease-Associated Single Nucleotide Polymorphism Using A Graphene Field Effect Transistor, Kandace Fung

CMC Senior Theses

Sickle Cell Disease (SCD) is a hereditary monogenic disorder that affects millions of people worldwide and is associated with symptoms such as stroke, lethargy, chronic anemia, and increased mortality. SCD can be quickly detected and diagnosed using a simple blood test as an infant, but as of now, there is currently limited treatment to cure an individual of sickle cell disease. Recently, there have been several promising developments in CRISPR-Cas-associated gene-editing therapeutics; however, there have been limitations in gene-editing efficiency monitoring, which if improved, could be beneficial to advancing CRISPR-based therapy, especially in SCD. The CRISPR-Chip, a three-terminal graphene-based field …

Exploring The Efficacy Of Natural Products In Alleviating Alzheimer’S Disease Using Animal Models, Prajakta Deshpande, Neha Gogia, Amit Singh

Biology Faculty Publications

Alzheimer’s disease (hereafter AD) is a progressive neurodegenerative disorder that affects the central nervous system. There are multiple factors that cause AD, viz., accumulation of extracellular Amyloid-beta 42 plaques, intracellular hyper-phosphorylated Tau tangles, generation of reactive oxygen species due to mitochondrial dysfunction and genetic mutations. The plaques and tau tangles trigger aberrant signaling, which eventually cause cell death of the neurons. As a result, there is shrinkage of brain, cognitive defects, behavioral and psychological problems. To date, there is no direct cure for AD. Thus, scientists have been testing various strategies like screening for the small inhibitor molecule library …

Biomass Yield In A Genetically Diverse Miscanthus Sinensis Germplasm Panel Evaluated At Five Locations Revealed Individuals With Exceptional Potential, Lindsay V. Clark, Maria S. Dwiyanti, Kossonou G. Anzoua, Joe E. Brummer, Bimal Kumar Ghimire, Katarzyna Glowacka, Megan Hall, Kweon Heo, Xiaoli Jin, Alexander E. Lipka, Junhua Peng, Toshihiko Yamada, Ji Hye Yoo, Chang Yeon Yu, Hua Zhao, Stephen P. Long, Erik J. Sacks

Department of Biochemistry: Faculty Publications

To breed improved biomass cultivars of Miscanthus ×giganteus, it will be necessary to select the highest‐yielding and best‐adapted genotypes of its parental species, Miscanthus sinensis and Miscanthus sacchariflorus. We phenotyped a diverse clonally propagated panel of 569 M. sinensis and nine natural diploid M. ×giganteus at one subtropical (Zhuji, China) and five temperate locations (Sapporo, Japan; Leamington, Ontario, Canada; Fort Collins, CO; Urbana, IL; and Chuncheon, Korea) for dry biomass yield and 14 yield‐component traits, in trials grown for 3 years. Notably, dry biomass yield of four Miscanthus accessions exceeded 80 Mg/ha in Zhuji, China, approaching …

An Arabidopsis Protoplast Isolation Method Reduces Cytosolic Acidification And Activation Of The Chloroplast Stress Sensor Sensitive To Freezing 2, Allison C. Barnes, Christian G. Elowsky, Rebecca Roston

Department of Biochemistry: Faculty Publications

Chloroplasts adapt to freezing and other abiotic stresses in part by modifying their membranes. One key-remodeling enzyme is SENSITIVE TO FREEZING2 (SFR2). SFR2 is unusual because it does not respond to initial cold stress or cold acclimation, instead it responds during freezing conditions in Arabidopsis. This response has been shown to be sensitive to cytosolic acidification. The unique lipid products of SFR2 have also been detected in response to non-freezing stresses, but what causes SFR2 to respond in these stresses is unknown. Here, we investigate protoplast isolation as a representative of wounding stress. We show that SFR2 oligogalactolipid products accumulate …

Application Of Short Tandem Target Mimic (Sttm) Technique For Functional Analysis Of Micro-Rna396 In Transgenic Poplar Trees, Surattana Boonsai

Dissertations, Master's Theses and Master's Reports

Short Tandem Target Mimic (STTM) is a commonly technique used for functional studies of a number of genes in several plant model systems. However, very little is known about application of STTM technique in tree species. In this study, STTM was applied to knock down microRNA396 (miR396) in transgenic poplar trees for the first time. STTM396 expression resulted in dramatic decrease in miR396 expression levels leading to taller plants with larger leaves and larger leaf cell size. Additionally, an expression analysis of growth regulating factor genes (GRFs) that are members of miR396 target gene family showed up-regulation of GRF07 gene …

Effective Knowledge Transfer During Biopharmaceutical Technology Transfer - How Well Do We Do It?, Martin J. Lipa, Paige Kane, Anne Greene

Articles

Author’s Note: This is the author’s submitted manuscript which has subsequently been published in the Journal of Validation Technology (www.ivtnetwork.com, Volume 25, Issue 4 – August 2019).

While knowledge management (KM) has been widely applied in other sectors, the international biopharmaceutical sector has struggled with the meaningful and sustained application of effective KM practices. This is evident even though KM has been highlighted in regulatory guidance for over 10 years, and the positive business impact of KM is well recognized in other sectors. This paper focuses on the topic of KM as applied to biopharmaceutical technology transfer, introducing …