Biotechnology Commons^™

Using Computational Fluid Dynamics And Optical Sensor Technology To Scale Cell Culture Platforms, Mandar Makwana

KGI Theses and Dissertations

Different cell culture vessels ranging from micro scale to laboratory scale to commercial scale play critical role in upstream process development for biologics manufacturing. Based on the mode of operation, cell culture vessels have different hydrodynamic environments, making it challenging to scale. Integrated approaches using computational tools supported by experimental studies can overcome these challenges. Computational Fluid Dynamics (CFD) is one such tool that can simulate hydrodynamics within the cell culture vessels and can provide insights at macro and micro-scale. Accuracy of a CFD model significantly depends on the fluid model and assumptions. Traditionally, simple two-equation fluid models were developed …

Evaluation Of Ionic Liquid Treatment As An Alternative To Chemical Fixation And Dehydration In The Preparation Of Biofilms For Scanning Electron Microscopy, Molly Shreve

Mahurin Honors College Capstone Experience/Thesis Projects

Scanning electron microscopes (SEM) give researchers the ability to observe very small features on biological specimens at the nanometer scale, which is a much higher resolution than what can be obtained with a light microscope. To be observed under SEM, unlike light microscopes, samples must be dry, conductive, and able to withstand electron bombardment. Preparation methods have been developed that work well for samples composed mainly of proteins and lipids, but these fail for samples that are predominantly carbohydrates, such as bacterial biofilms. When carbohydrates of these biofilms are dehydrated, they tend to shrink and collapse, and their normal hydrated …

Mechanisms Of Stimulation By Intestinal Endothelial Cells During Hiv Infection Of Activated Cd4+ T Cells, Anding Shen, Timothy Kim, Sarah Logan, Zihan Zhang

Summer Research

Under antiretroviral therapy, the viral persistence of HIV is enabled by latent viral reservoirs, particularly in resting CD4+ T cells. Recent studies reveal interactions between endothelial cells and T cells allowing for latent infection of resting and activated CD4+ T cells. We focused our investigation on the stimulation of activated T cells by intestinal endothelial cells (IECs), due to the prominent gut pathology of HIV and predominance of an activated T cell phenotype in the gut. While prior research shows the involvement of soluble factors like IL-6 in stimulation of resting T cells, our results from transwell cultures suggest the …

Understanding The Non-Canonical Function Of Ezh1/2 In Gene Activation, Visakuo Tsurh, Longxia Xu, Xiaobing Shi

Summer Research

Enhancers of Zeste homolog proteins (EZH1 and EZH2) play a critical role in regulating gene transcription during development. The typical function of EZH2 and its homolog EZH1 is catalyzing the trimethylation of histone three at lysine 27 (H3K27) to repress gene expression through the PRC2 complex. In cancer, EZH2 is often over-expressed or mutated, leading to the silencing of tumor suppressor genes and a worse prognosis of the disease. Research on EZH1 and EZH2 in cancer has focused on their canonical function and has culminated in therapies that target their enzymatic activity. However, studies have found that drugs that inhibit …

The Effects Of Exogenously Applied Antioxidants On Plant Growth And Resilience, Aline Rodrigues De Queiroz, Connor Hines, Jeremy Brown, Seema Sahay, Jithesh Vijayan, Julie M. Stone, Nate Bickford, Melissa Wuellner, Katarzyna Glowacka, Nicole R. Buan, Rebecca Roston

Department of Biochemistry: Faculty Publications

Plant growth and resilience require balancing an inherently oxidative metabolism with powerful antioxidant systems that help maintain homeostasis. When the environment changes, reactive oxygen species are potent indicators of that change, allowing adaptation through re-balancing metabolism and antioxidant systems. A large body of evidence supports the use of exogenously applied antioxidants to improve both plant growth and their resilience to stress. Notably, some phenotypic effects are similar upon the application of chemically diverse antioxidants, while others are distinct. In this review, we analyze research from antioxidant treatment experiments and highlight the similarities in their practical applications and their effects on …

Preclinical Assessment Of Psychoplastogens For Treatment Of Psychiatric Symptoms And Neuroplasticity Deficits In Huntington Disease, Nicholas Skiados

Electronic Theses and Dissertations, 2020-2023

Huntington disease (HD) is a neurodegenerative disorder caused by a CAG trinucleotide repeat expansion in the huntingtin (HTT) gene. HD manifests as a triad of psychiatric, cognitive, and motor symptoms, the latter of which defines symptomatic onset of the disease. Psychiatric symptoms including aggression and depression often emerge decades prior to motor symptom onset. Psychiatric disorders are closely linked with dysregulated neural plasticity, the ability of the brain to form new synaptic connections. Neural plasticity is impaired in HD, potentially playing a role in psychiatric symptoms. However, the association between plasticity and psychopathology in HD has yet to be thoroughly …

Genetic Control Of Photoprotection And Photosystem Ii Operating Efficiency In Plants, Seema Sahay, Marcin Grzybowski, James Schnable, Katarzyna Głowacka

Department of Biochemistry: Faculty Publications

• Photoprotection against excess light via nonphotochemical quenching (NPQ) is indispensable for plant survival. However, slow NPQ relaxation under low light conditions can decrease yield of field-grown crops up to 40%.

• Using semi-high-throughput assay, we quantified the kinetics of NPQ and photosystem II operating efficiency (ΦPSII) in a replicated field trial of more than 700 maize (Zea mays) genotypes across 2 yr. Parametrized kinetics data were used to conduct genome-wide association studies.

• For six candidate genes involved in NPQ and ΦPSII kinetics in maize the loss of function alleles of orthologous genes in Arabidopsis (Arabidopsis thaliana) …

Plastic Recognition And Electrogenic Uniport Translocation Of 1st-, 2nd-, And 3rd-Row Transition And Post-Transition Metals By Primary-Active Transmembrane P1b-2-Type Atpase Pumps, Sameera S. Abeyrathna, Nisansala S. Abeyrathna, Priyanka Basak, Gordon W. Irvine, Limei Zhang, Gabriele Meloni

Department of Biochemistry: Faculty Publications

Transmembrane P-type ATPase pumps catalyze the extrusion of transition metal ions across cellular lipid membranes to maintain essential cellular metal homeostasis and detoxify toxic metals. Zn()-pumps of the P-type subclass, in addition to Zn , select diverse metals (Pb, Cdand Hg) at their transmembrane binding site and feature promiscuous metal-dependent ATP hydrolysis in the presence of these metals. Yet, a comprehensive understanding of the transport of these metals, their relative translocation rates, and transport mechanism remain elusive. We developed a platform for the characterization of primary-active Zn()-pumps in proteoliposomes to study metal selectivity, translocation events and transport mechanism in real-time, …

Genome-Wide Investigation Of Snrk2 Gene Family In Two Jute Species: Corchorus Olitorius And Corchorus Capsularis, Borhan Ahmed, Fakhrul Hasan, Anika Tabassum, Rasel Ahmed, Rajnee Hassan, Ruhul Amin, Mobashwer Alam

Department of Biochemistry: Faculty Publications

Background Sucrose non-fermenting-1 (SNF1)-related protein kinase 2 (SnRK2), a plant-specifc serine/threonine kinase family, is associated with metabolic responses, including abscisic acid signaling under biotic and abiotic stresses. So far, no information on a genome-wide investigation and stress-mediated expression profling of jute SnRK2 is available. Recent whole-genome sequencing of two Corchorus species prompted to identify and characterize this SnRK2 gene family.

Result We identifed seven SnRK2 genes of each of Corchorus olitorius (Co) and C. capsularis (Cc) genomes, with similar physico-molecular properties and sub-group patterns of other models and related crops. In both species, the SnRK2 gene …

Structural Basis Of Dna Binding By The Whib-Like Transcription Factor Whib3 In Mycobacterium Tuberculosis, Tao Wan, Magdaléna Horová, Vimmy Khetrapal, Shanren Li, Camden Jones, Andrew Schacht, Xinghui Sun, Limei Zhang

Department of Biochemistry: Faculty Publications

Mycobacterium tuberculosis (Mtb) WhiB3 is an iron–sulfur cluster-containing transcription factor belonging to a subclass of the WhiB-Like (Wbl) family that is widely distributed in the phylum Actinobacteria. WhiB3 plays a crucial role in the survival and pathogenesis of Mtb. It binds to the conserved region 4 of the principal sigma factor (σ^A₄) in the RNA polymerase holoenzyme to regulate gene expression like other known Wbl proteins in Mtb. However, the structural basis of how WhiB3 coordinates with σ^A₄ to bind DNA and regulate transcription is unclear. Here we determined crystal structures …

Factors Influencing Instructors’ Adoption And Continued Use Of Computing Science Technologies: A Case Study In The Context Of Cell Collective, Changsoo Song, Resa M. Helikar, Wendy M. Smith, Tomáš Helikar

Department of Biochemistry: Faculty Publications

Acquiring computational modeling and simulation skills has become ever more critical for students in life sciences courses at the secondary and tertiary levels. Many modeling and simulation tools have been created to help instructors nurture those skills in their classrooms. Understanding the factors that may motivate instructors to use such tools is crucial to improve students’ learning, especially for having authentic modeling and simulation learning experiences. This study designed and tested a decomposed technology acceptance model in which the perceived usefulness and perceived ease of use constructs are split between the teaching and learning sides of the technology to examine …

Akkermansia Muciniphila And Its Membrane Protein Ameliorates Intestinal Inflammatory Stress And Promotes Epithelial Wound Healing Via Crebh And Mir‑143/145, Henry Wade, Kaichao Pan, Qihua Duan, Szczepan Kaluzny, Ekta Pandey, Linda Fatumoju, Viswanathan Saraswathi, Rongxue Wu, Edward N. Harris, Qiaozhu Su

Department of Biochemistry: Faculty Publications

Background The intestinal epithelial barrier is the interface for interaction between gut microbiota and host metabolic systems. Akkermansia muciniphila (A. muciniphila) is a key player in the colonic microbiota that resides in the mucus layer, whose abundance is selectively decreased in the faecal microbiota of inflammatory bowel disease (IBD) patients. This study aims to investigate the regulatory mechanism among A. muciniphila, a transcription factor cAMPresponsive element-binding protein H (CREBH), and microRNA-143/145 (miR-143/145) in intestinal inflammatory stress, gut barrier integrity and epithelial regeneration.

Methods A novel mouse model with increased colonization of A muciniphila in the intestine of CREBH knockout …

Advancements In Computational Modelling Of Biological Systems: Seventh Annual Sysmod Meeting, Bhanwar Lal Puniya, Andreas Dräger

Department of Biochemistry: Faculty Publications

The Computational Modelling of Systems Biology (SysMod) Community of Special Interest (COSI) convenes annually at the Intelligent Systems for Molecular Biology (ISMB) conference to facilitate knowledge dissemination and exchange of research findings on systems modelling from interdisciplinary domains. The SysMod meeting 2022 was held in a hybrid mode in Madison, Wisconsin, spanning a 1-day duration centred on modelling techniques, applications, and single-cell technology implementations. The meeting showcased innovative approaches to modelling biological systems using cell-specific and multiscale modelling, multiomics data integration, and novel tools to develop systems models using single-cell and multiomics technology. The meeting also recognized outstanding research by …

Fusarium Graminearum Effector Fgnls1 Targets Plant Nuclei To Induce Wheat Head Blight, Guixia Hao, Todd A. Naumann, Hui Chen, Guihua Bai, Susan Mccormick, Hye-Seon Kim, Bin Tian, Harold N. Trick, Michael J. Naldrett, Robert Proctor

Nebraska Center for Biotechnology: Faculty and Staff Publications

Fusarium head blight (FHB) caused by Fusarium graminearum is one of the most devastating diseases of wheat and barley worldwide. Effectors suppress host immunity and promote disease development. The genome of F. graminearum contains hundreds of effectors with unknown function. Therefore, investigations of the functions of these effectors will facilitate developing novel strategies to enhance wheat resistance to FHB. We characterized a F. graminearum effector, FgNls1, containing a signal peptide and multiple eukaryotic nuclear localization signals. A fusion protein of green fluorescent protein and FgNls1 accumulated in plant cell nucleiwhen transiently expressed in Nicotiana benthamiana. FgNls1 suppressed …

The Role Of Myocardin In The Progression Of Non-Small Cell Lung Cancer, Soromidayo Akinsiku

Biotechnology Theses

Lung cancer is the leading cause of cancer-related mortality in the world and NSCLC accounts for 85% of all lung cancer cases. The mainstay of treatment for patients with stage I, II and IIIA NSCLC is surgery, followed by post-operative cisplatin-based chemotherapy. Additional adjuvant therapy involving targeted tyrosine kinase inhibitors has been in use, however even for the targeted therapy, resistance eventually develops. Therefore, there is a need for identifying novel targets for this life-threatening disease. Given that preliminary studies in Ikebe lab revealed that myocardin knockdown significantly promoted caspase-3 degradation, in this study, using myocardin siRNA, we investigated the …

Biomechanics Of Mantis Prey Capture, Danielle S. Taylor

Honors Theses

Mantis species have a variety of different morphologies, so do the extreme forms of mantis limbs trade prey catching capability for camouflage? We hypothesize that some extreme forms of mantis limbs that are associated with cryptic species may be associated with a tradeoff of the capability of those limbs. Previous research has developed 2D morphologies of several hundred species of mantises. We are creating a 3D morphology by using micro dissection, micro CT imaging to construct our 3D biomechanical model. We found the attachment points of the ligaments and muscles from a Tenodera forearm and have constructed a 2D biomechanical …

Efficient Text Bounding Box Identification Using Mask R-Cnn: Case Of Thai Documents, Phanthakan Kiatphaisansophon

Chulalongkorn University Theses and Dissertations (Chula ETD)

Text detection is a fundamental task in computer vision, particularly for Optical Character Recognition (OCR) application. This research focuses on text detection part of the OCR. Our previous text detection model, CRAFT (Character-Region Awareness For Text detection) has shown promising outcomes in bounding box identification. Nevertheless, it encounters issues associated with post-processing and multiline text. The post-processing related to the necessity to reconfigure the model when new documents are introduced, resulting in inefficiencies and complexities. Also the CRAFT model tends to blend bounding boxes from consecutive lines, introducing errors and impacting the accuracy in the text recognition part of the …

Analysis Of Biologically Effective Dose For Retroactive Yttrium-90 Trans-Arterial Radioembolization Treatment Optimization, Mj Lindsey

CMC Senior Theses

Trans-arterial radioembolization (TARE) is a protracted modality of radiation therapy where radionuclides labeled with Yttrium-90 (⁹⁰Y) are inserted inside a patient's hepatic artery to treat hepatocellular carcinoma (HCC). While TARE has been shown to be a clinically effective and safe treatment, there is little understanding of the radiobiological relationship between absorbed dose and tissue response, and thus there is no dosimetric standard for treatment planning. The Biologically Effective Dose (BED) formalism, derived from the Linear-Quadratic model of radiobiology, is used to weigh the absorbed dose by the time pattern of delivery. BED is a virtual dose that can …

Role Of Histone Deacetylases In Regulation Of Macrophage Phagocytosis Of Mycobacterium Tuberculosis, Benjamin Esiesimua

Biotechnology Theses

Tuberculosis (TB), caused by Mycobacterium tuberculosis (Mtb), remains a global health emergency. Macrophages are critical in protection against TB by phagocytosis of Mtb. Although histone deacetylases (HDACs), are critical in macrophage differentiations, their roles in macrophage phagocytosis of Mtb remain unexplored. To explore this, I first established human monocyte derived macrophages (MDMs) phagocytosis of red zymosan beads. MDM phagocytosed zymosan beads at 37°C in a dose dependent manner in contrast to suppressed phagocytosis by the MDMs incubated with beads at 4°C or in the presence of cytochalasin D, actin polymerization inhibitor. Incubation of MDMs with 11 zinc-dependent HDAC inhibitors did …

Development Of Novel Detection Methods For Food Safety Using State-Of-The-Art Technology Platforms, Prae Noppakuadrittidej

Chulalongkorn University Theses and Dissertations (Chula ETD)

Foodborne illness is a significant global public health concern, impacting both the economy and human health. These illnesses are caused by bacteria, viruses, parasites, toxins, and chemicals. This research developed two technology platforms for Bacillus cereus and tropane alkaloid compounds detections. First, traditional detection methods for B. cereus have limitations in long analysis time, and dependence on analysis and interpretation by skilled personnel. Therefore, this study introduces a robust detection method using peptide nucleic acids (PNAs) bearing with a prolyl-2-aminocyclopentanecarboxylic acid (ACPC) backbone as probes in a bead array method. PNAs with groEL, motB, and 16S rRNA sequences were covalently …