Cell and Developmental Biology Commons^™

Spiral Ganglion Degeneration And Hearing Loss As A Consequence Of Satellite Cell Death In Saposin B-Deficient Mice, Omar Akil, Ying Sun, Sarath Vijayakumar, Wujuan Zhang, Tiffany Ku, Chi-Kyou Lee, Sherri Jones, Gregory A. Grabowski, Lawrence R. Lustig

School of Veterinary and Biomedical Sciences: Faculty Publications

Saposin B (Sap B) is an essential activator protein for arylsulfatase A in the hydrolysis of sulfatide, a lipid component of myelin. To study Sap B’s role in hearing and balance, a Sap B-deficient (B^-/-) mouse was evaluated. At both light and electron microscopy (EM) levels, inclusion body accumulation was seen in satellite cells surrounding spiral ganglion (SG) neurons from postnatal month 1 onward, progressing into large vacuoles preceding satellite cell degeneration, and followed by SG degeneration. EM also revealed reduced or absent myelin sheaths in SG neurons from postnatal month 8 onwards. Hearing loss was initially seen …

Pseudorabies Virus Fast Axonal Transport Occurs By A Pus9- Independent Mechanism, Gina R. Daniel, Patricia J. Sollars, Gary E. Pickard, Gregory A. Smith

School of Veterinary and Biomedical Sciences: Faculty Publications

Reactivation from latency results in transmission of neurotropic herpesviruses from the nervous system to body surfaces, referred to as anterograde axonal trafficking. The virus-encoded protein pUS9 promotes axonal dissemination by sorting virus particles into axons, but whether it is also an effector of fast axonal transport within axons is unknown. To determine the role of pUS9 in anterograde trafficking, we analyzed the axonal transport of pseudorabies virus in the presence and absence of pUS9.

Haemorrhagic Colitis Associated With Enterohaemorrhagic Escherichia Coli O165:H25 Infection In A Yearling Feedlot Heifer, Rodney A. Moxley, Zachary R. Stromberg, Gentry L. Lewis, John D. Loy, Bruce W. Brodersen, Isha R. Patel, Jayanthi Gangiredla

School of Veterinary and Biomedical Sciences: Faculty Publications

Introduction: Enterohaemorrhagic Escherichia coli (EHEC) cause haemorrhagic colitis and haemolytic uraemic syndrome in humans. Although EHEC infection typically results in haemorrhagic colitis in all ages of human patients, in cattle it is usually limited to 1- to 5-week-old nursing calves.

Case Presentation: A 1-year-old feedlot beef heifer was moribund with neurological signs and bloody diarrhoea. At necropsy, the colonic mucosa contained multiple grossly visible haemorrhagic erosions, each measuring <1 mm in diameter. Histologically, foci corresponding to the gross erosions had E. coli O165 antigen-positive bacterial rods adherent to the apical surfaces of degenerate and necrotic colonic mucosal epithelial cells in association with attaching and effacing lesions, and also within cytoplasmic vacuoles in …

Characterization Of Newly Revealed Sequences In The Infectious Myonecrosis Virus Genome In Litopenaeus Vannamei, Duan S. Loy, Sijun Liu, Mark A. Mogler, J. Dustin Loy, Bradley J. Blitvich, Lyric C. Bartholomay

School of Veterinary and Biomedical Sciences: Faculty Publications

Infectious myonecrosis virus (IMNV) causes significant economic losses in farmed shrimp, where associated mortality in ponds can reach 70%. To explore host/pathogen interactions, a next-generation sequencing approach using lymphoid organ tissue from IMNV-infected Litopenaeus vannamei shrimp was conducted. Preliminary sequence assembly of just the virus showed that there were at least an additional 639 bp at the 5′ terminus and 23 nt at the 3′ terminus as compared with the original description of the IMNV genome (7561 nt). Northern blot and reverse transcription-PCR analysis confirmed the presence of novel sequence at both ends of the genome. Using 5′ RACE, an …

Mri-Compatible Bioreactors And Methods Of Using, Shadi Othman, Huihui Xu, Karin Wartella, Vahid Khalilzad-Sharghi, Ian Bargar

School of Veterinary and Biomedical Sciences: Faculty Publications

This disclosure describes a MRI-compatible bioreactor that allows a biological sample to be imaged in culture without disrupting or compromising the culture.

Effects Of Acetyl Salicyclic Acid On Drosophila Models Of Cancer-Inflammation, Roma Rajwani

Dissertations and Theses

Chronic inflammation is a fundamental condition underlying many human disease pathologies including diabetes, cancer, and neurodegeneration. Indeed, one of the most commonly-used anti-inflammatory drugs, aspirin, has been shown to be effective in the treatment of many cancers. To understand the mechanistic linkages between inflammation and cancer, we used Drosophila models in which hyperactive immune signaling causes overproliferation of the hematopoietic system leading to the production of inflammatory blood tumors. In mutant animals, hematopoietic cells divide uncontrollably and exhibit pre-mature differentiation giving rise to large sticky cells. Some mutant cells infiltrate and adhere to cells of the fat body. The fat …

Characterization Of The Interaction Between Ftsz-Ring Stabilizer Zapc And The Conserved Protease Clpxp In E. Coli Cytokinesis, Andrea Cardenas Arevalo

Dissertations and Theses

Cell division in bacteria requires the assembly of a macromolecular protein machinery at midcell that is spatiotemporally regulated during the bacterial cell cycle. Central to the process of division is the assembly of a cytokinetic ring-like structure, termed the Z-ring, formed by polymers of the essential tubulin-homolog FtsZ. Polymerization of FtsZ is regulated by a number of proteins that bind FtsZ and regulate its assembly/disassembly mechanisms. Despite being extensively studied, the molecular nature of the diverse protein-protein interactions that regulate the assembly of FtsZ during early stages of division is not well understood. Here, I focus on an FtsZ-ring stabilizer …

Bridging In Vi Tro & In S Il Ico Techniques: Uncovering The Structural Characteristics And Light-Dependent Modifications Of The Light-Response Btb Proteins In Arabidopsis Thaliana, Nicholas Reitz

Masters Theses

The conjugation of the NEDD8 protein has been found to cause a wide range of changes to the overall function of a protein. The addition of NEDD8 to a conserved lysine on Cul proteins has been found to provide Cul with increased flexibility, while the transcription factor protein p53 was prevented from binding to DNA after NEDD8 conjugation. Using Arabidopsis thaliana, we investigated the potential conjugation of NEDD8 to the AtLRB1 and AtLRB2 proteins, which are both members of Cul3-RING E3 Ubiquitin Ligase complexes, using both in vitro and in silico techniques. The AtLRBs are negative regulators of the red …

Gene Expression And Alzheimer's Disease: Evaluation Of Gene Expression Patterns In Brain And Blood For An Alzheimer's Disease Mouse Model, Amanda Hazy

Senior Honors Theses

Previous studies have established a causative role for altered gene expression in development of Alzheimer’s disease (AD). These changes can be affected by methylation and miRNA regulation. In this study, expression of miRNA known to change methylation status in AD was assessed by qPCR. Genome-wide expression changes were determined by RNA-sequencing of mRNA from hippocampus and blood of control and AD mice. The qPCR data showed significantly increased expression of Mir 17 in AD, and sequencing data revealed 230 genes in hippocampus, 58 genes in blood, and 8 overlapping genes showing significant differential expression (p value ≤ 0.05). Expression data …

Detection And Tracking Of T Cells In Time-Lapse Imaging, Cody Arbuckle, Milton L. Greenberg, Erik J. Linstead

Mathematics, Physics, and Computer Science Faculty Articles and Research

The effective classification and tracking of cells obtained from modern staining techniques has significant limitations due to the necessity of having to train and utilize a human expert in the field who must manually identify each cell in each slide. Often times these slides are filled with noise cells that are not of particular interest to the researcher. The use of computational methods has the ability to effectively and efficiently enhance image quality, as well as identify and track target cell types over large data sets. Here we present a computational approach to the in vitro tracking of T cells …

Dose-Dependent Atp Depletion And Cancer Cell Death Following Calcium Electroporation, Relative Effect Of Calcium Concentration And Electric Field Strength, Emilie Louise Hansen, Esin Bengisu Sozer, Stefania Romeo, Stine Krog Frandsen, P. Thomas Vernier, Julie Gehl

Bioelectrics Publications

Background: Electroporation, a method for increasing the permeability of membranes to ions and small molecules, is used in the clinic with chemotherapeutic drugs for cancer treatment (electrochemotherapy). Electroporation with calcium causes ATP (adenosine triphosphate) depletion and cancer cell death and could be a novel cancer treatment. This study aims at understanding the relationship between applied electric field, calcium concentration, ATP depletion and efficacy. Methods: In three human cell lines — H69 (small-cell lung cancer), SW780 (bladder cancer), and U937 (leukaemia), viability was determined after treatment with 1, 3, or 5 mM calcium and eight 99 μs pulses with 0.8, 1.0, …

Presence And Localization Of Pro-And Mature Forms Of Biglycan And Decorin In Human Costal Cartilage Derived From Chest Wall Deformities, Anthony Asmar, A. Werner, R. E. Kelly Jr., A. Fecteau, Michael W. Stacey

Bioelectrics Publications

Costal cartilage is a type of hyaline cartilage that forms rod-like structures that connect the ribs to the sternum. The most common chest wall deformities, pectus excavatum and pectus carinatum involved efective costal cartilage resulting in sternal displacement. Costal cartilage is not widely studied leaving little insight into possible factors involved in the pathogenesis of these pectus deformities. This study focused on the presence and distribution of two important regulators of collagen fibrillogenesis and organization, biglycan and decorin. Immunohistochemical analysis of transverse cross sections of normal and deformed costal cartilage revealed that biglycan and decorin mainly localized in the territorial …

Utilizing Galectin 3 As A Marker Of Ruptured Vesicles In Vivo, Jonathan London

Master's Theses

Ruptured vesicles have been associated with pathological forms of α-synuclein during the development of PD. Galectin 3 (Gal3) is readily recruited to these ruptured vesicles, making it a potentially useful marker of vesicle rupture in PD and other diseases associated with vesicle rupture and lysosomal dysfunction. We were able to visualize and quantify the differences in Gal3 levels during infection with pathological α-synuclein in the rat brain. Having significantly high basal levels of Gal3, we found that the differentiated monocytes showed a nearly100 percent increase in intercellular levels of Gal3 after treatment with pathological α-synuclein. By expressing only the Carbohydrate …

Expression And Characterization Of Ecm14, A Metallocarboxypeptidase From Saccharomyces Cerevisiae, Matthew James Schott

Master's Theses

Metallocarboxypeptidases are found in most organisms and function in the digestion and maturation of proteins. Ecm14 is a putative metallocarboxypeptidase found in the yeast Saccharomyces cerevisiae vacuole. The function of Ecm14 as an enzyme has been unclear due to the presence of active site amino acids not typically found in metallocarboxypeptidases, suggesting either no enzymatic activity or a unique mechanism. In order to investigate the enzymatic mechanism of Ecm14, expression of histidine-tagged Ecm14 protein was attempted in human HEK293T cell culture, S. cerevisiae, and baculovirus expression systems. No expression was detected in HEK293T cells in preliminary experiments. Expression in the …

Evaluation Of The Developmental Toxicity Of Lead In The Danio Rerio Body, Nicole M. Roy, Sarah Dewolf, Bruno Carneiro

Biology Faculty Publications

Lead has been utilized throughout history and is widely distributed and mobilized globally. Although lead in the environment has been somewhat mitigated, the nature of lead and its extensive uses in the past prohibit it from being completely absent from our environment and exposure to lead is still a public health concern. Most studies regarding lead toxicity have focused on the brain. However, little is found in the literature on the effects of lead in other tissues. Here, we utilize the zebrafish model system to investigate effects of lead exposure during early developmental time windows at 24, 48 and 72 …

Immunotherapy Of Cancer: Reprogramming Tumor/Immune Cellular Crosstalk To Improve Anti-Tumor Efficacy, Kyle K. Payne

Theses and Dissertations

Immunotherapy of cancer has been shown to be promising in prolonging patient survival. However, complete elimination of cancer and life-long relapse-free survival remain to be major challenge for anti-cancer therapeutics. We have previously reported that ex vivo reprogramming of tumor-sensitized immune cells by bryostatin 1/ionomycin (B/I) and the gamma-chain (γ-c) cytokines IL-2, IL-7, and IL-15 resulted in the generation of memory T cells as well as CD25+ NKT cells and CD25+ NK cells. Adoptive cellular therapy (ACT) utilizing these reprogrammed immune cells protected FVBN202 mice from tumor challenge, and overcame the suppressive functions of myeloid-derived suppressor cells (MDSCs). We then …

The Role Of Srsf3 In Control Of Alternative Splicing Of Cpeb2 In Triple Negative Breast Cancer, Brian P. Griffin

Theses and Dissertations

In the presented study, we identified that SRSF3 controls the alternative splicing of CPEB2 and consequently promotes a metastatic phenotype in triple negative breast cancer (TNBC). TNBC causes thousands of deaths annually, frequently due to a lack of effective treatments and a high rate of metastasis in patients. Alternative splicing has been found to be dysregulated in numerous cancers, while splicing factors such as SRSF3 are variably expressed. In this study we performed a siRNA panel to screen potential splicing factors, then used specific siRNA to study the effect of its knockdown on cellular function. These results showed that SRSF3 …

Pemetrexed, A Modulator Of Amp-Activated Kinase Signaling And An Inhibitor Of Wild Type And Mutant P53, Stuti Agarwal

Theses and Dissertations

New drug discoveries and new approaches towards diagnosis and treatment have improved cancer therapeutics remarkably. One of the most influential and effective discoveries in the field of cancer therapeutics was antimetabolites, such as the antifolates. The interest in antifolates increased as some of the antifolates showed responses in cancers, such as mesothelioma, leukemia, and breast cancers. When pemetrexed (PTX) was discovered, our laboratory had established that the primary mechanism of action of pemetrexed is to inhibit thymidylate 22 synthase (TS) (E. Taylor et al., 1992). Preclinical studies have shown that PTX has a broad range of antitumor activity in human …

Interaction Between Atm Kinase And P53 In Determining Glioma Radiosensitivity, Syed F. Ahmad

Theses and Dissertations

Glioblastoma multiforme (GBM) is the most common primary brain tumor. Studies have shown that targeting the DNA damage response can sensitize cancer cells to DNA damaging agents. Ataxia telangiectasia mutated (ATM) is involved in signaling DNA double strand breaks. Our group has previously shown that ATM inhibitors (ATMi) sensitize GBM cells and tumors to ionizing radiation. This effect is greater when the tumor suppressor p53 is mutated.

The goals of this work include validation of a new ATM inhibitor, AZ32, and elucidation of how ATMi and p53 status interact to promote cell death after radiation. We propose that ATMi and …

Nitric Oxide Synthase Activity And Its Modulation In The Treatment Of Colorectal Cancer, Asim Alam

Theses and Dissertations

The American Cancer Society estimates more than 141,000 new cases of and about 50,000 deaths from colorectal cancer every year. Treatment options include surgery, radiation therapy and targeted therapies such as anti-angiogenics. However, no therapies address the key driving factor of colorectal cancer: inflammation. It is well known that chronic inflammatory conditions such as Crohn’s Disease, ulcerative colitis, diabetes, obesity and cigarette smoking all elevate the risk of developing colorectal cancer. One of the hallmarks of chronic inflammation is the elevated levels of reactive oxygen/nitrogen species (ROS/RNS). A primary source of these ROS/RNS is uncoupled Nitric Oxide Synthase (NOS). Under …