Digital Commons Network^™

Transcriptional Dysregulation In Interneurons Causes Altered Modulation Of Hippocampal Synaptic Transmission And Circuit Function By The Dopamine System, Lillian J. Brady

All ETDs from UAB

Genetic deletion of the transcriptional coactivator PGC-1α leads to transcriptional dysregulation in interneurons, and transcriptional dysregulation in interneurons lead to changes in hippocampal synaptic transmission and circuit function. Hippocampal circuit activity and synaptic transmission alterations have implications for symptoms of cognitive impairment in neurological disorders like schizophrenia. Modulation of dopamine receptor activity through pharmacologic application of haloperidol and the specific dopamine D4 receptor antagonist L-745,870 in PGC-1α-/- mice leads to altered effects on inhibitory/excitatory synaptic transmission balance, circuit function, and innate hippocampal dependent nesting behavior. These are key aspects underlying hippocampus dependent cognitive impairment. Specifically, bath application of haloperidol restores …

Immunobiology Of The Mucosal Response To Helicobacter Pylori In Children, Kyle Brawner

All ETDs from UAB

Helicobacter pylori colonizes the stomach of approximately one half of the world’s population. However, the vast majority of infected persons remain asymptomatic. The degree of H. pylori-induced gastric inflammation depends on a complex interaction of several factors. In this dissertation, we focus on age as a determinant of gastritis severity. Children are known to respond to H. pylori infection with a stronger T regulatory cell profile compared with infected adults, yet the reason remains unknown. The influence of the microbiota on, and the communication among, cells of the innate immune system shape the nature of the adaptive immune response to …

Interplay Of The Circadian Clock And Growth Hormone Signaling In The Heart, Ryan Darrell Berry

All ETDs from UAB

Growth hormone (GH) signaling plays a critical role in postnatal development and the details of its release, regulation, and influence on the development of a number of organs has been well studied. GH is released from the anterior pituitary in a pulsatile fashion with pulse amplitude being greater at night than during the day in a number of species, including humans. Once in circulation, GH binds to the GH receptor (GHR) on target tissues throughout the body altering gene expression and stimulating the local production of insulin-like growth factor 1 (IGF-1). IGF-1 primarily acts in an autocrine and paracrine fashion. …

Optimizing The Non-Ablative Conditioning Regimen For Bone Marrow Transplantation Using A Murine Model Of Sickle Cell Disease, Divya Devadasan

All ETDs from UAB

In this dissertation, we sought to determine if a sub-myeloablative conditioning regimen followed by bone marrow transplantation/hematopoietic stem cell transplantation (HSCT) using syngeneic donor cells could facilitate donor chimerism using a clinically relevant mouse model of sickle cell disease (SCD). This pre-clinical study determined the feasibility of using sub-myeloablative conditioning in the context of autologous HSCT. The prospect of using less-intense conditioning is particularly appealing in the era of modern gene-editing technologies since its extension to use in humans will allow for HSCT to be less toxic and hence be applicable in all patients. We showed that using a sub-myeloablative …

O-Acetyltransferase Functionality Mediates Pneumococcal Capsular Diversity, Brady Spencer

All ETDs from UAB

Streptococcus pneumoniae (pneumococcus) is an important human pathogen that asymptomatically colonizes the nasopharynx but can disseminate opportunistically to cause diseases such as otitis media, pneumonia, septicemia, or meningitis. The pneumo-coccal capsular polysaccharide is necessary to prevent opsonophagocytosis as nonencap-sulated pneumococci are largely avirulent. Currently, 98 antigenically distinct capsule types (serotypes) have been identified, which vary in virulence. O-acetylation, a subtle capsular modification, contributes to pneumococcal capsular diversity and modulates capsular physicochemical properties and interaction with innate or vaccine immunity. Replacement serotypes (and future vaccine types) 33F and 15B have O-acetylated capsules due to membrane-bound O-acetyltransferases (MOATs). We hypothesize that future …

The Role Of Klotho In Adult Neurogenesis, Ann Laszczyk

All ETDs from UAB

Klotho is an anti-aging protein that when upregulated, extends lifespan and enhances cognition. Klotho knockout mice age prematurely and die by ~8 weeks of life after developing cognitive impairment. Klotho function outside of the brain is well established but the protein’s action within the brain remains unknown. As klotho regulates peripheral stem cell populations, I sought to determine whether klotho contributes to cognition through a role in the maintenance of the adult hippocampal neurogenic niche. I examined adult mouse neurogenesis using global klotho-deficiency or global klotho overexpression models. Klotho knockout brains show reduced progenitor proliferation and granule cell production while …

Alterations In Ubc9 Substrate Specificity Affect The Cellular Response To Dna Damage, Joshua Enyeribe Onuiri

All ETDs from UAB

This dissertation seeks to uncover the impact of a post-translational modification, the small ubiquitin-related modifier (SUMO), on the cell response to DNA damage. Covalent modification of cellular proteins by SUMO occurs through an enzymatic cascade involving an E1 activating enzyme, an E2 conjugating enzyme, and a ligase (E3). Eukaryotic cells possess a single E1 and a single E2 for SUMO conjugation. The E2 for SUMO, Ubc9, is able to recognize and modify substrates on its own, although substrate recognition and modification are enhanced in the presence of the E3. Two independent genetic screens carried out in the budding yeast Saccharomyces …

The Role Of Macrophages In Pressure-Overload Induced Cardiac Remodeling, Bindiya Dinesh Patel

All ETDs from UAB

Cardiac pressure-overload is a state of increased hemodynamic load and is initially characterized by concentric left ventricular (LV) hypertrophy, but can transition to eccentric hypertrophy and adverse LV remodeling leading to heart failure (HF). Cardiac remodeling during pressure-overload is accompanied by dysregulated collagen deposition, cardiomyocyte hypertrophy, and increased inflammatory signaling. Both clinical and preclinical studies have documented increased expression of inflammatory cytokines and leukocyte infiltration in the pressure-overloaded heart, indicating inflammatory mechanisms are active during cardiac remodeling. Despite the well-known link between leukocyte infiltration and exacerbated tissue injury, the role of innate immune cells, particularly monocytes, macrophages and dendritic cells, …

Investigating Phenotypic Severity Associated With Sister Chromatid Cohesion Defects In Human Disease, Stefanie Marie Percival

All ETDs from UAB

INVESTIGATING PHENOTYPIC SEVERITY ASSOCIATED WITH SISTER CHROMATID COHESION DEFECTS IN HUMAN DISEASE STEFANIE M. PERCIVAL GRADUATE BIOMEDICAL SCIENCES ABSTRACT Sister chromatid cohesion (SCC) is a process that utilizes a proteinaceous ring, cohesin, for accurate chromosome segregation. An essential process in S phase termed cohesion establishment is necessary to stabilize cohesin rings around sister chromatids. Mutations in establishment of cohesion homolog 2 (ESCO2), a protein essential for cohesion establishment, cause a developmental disorder called Roberts Syndrome (RBS). Cytogenetic analysis in patients reveals heterochromatic repulsion (HR), a centromeric puffing, indicative of cohesion defects. The severity of phenotypes varies from preterm lethal to …

Elucidating The Physiologic And Prognostic Significance Of N-Myc And Stat Interactor Using Models Of Mammary Development And Metastasis, Hawley Christine Pruitt

All ETDs from UAB

Distant metastasis of breast carcinoma reduces the five year survival rate of patients from 90% to a dismal 25%. Although the metastatic cascade has been extensively studied for decades, players that influence the progression of metastatic disease remain elusive. N-MYC and STAT Interactor (NMI) is a gene previously demonstrated by Devine et. al. to be downregulated with metastatic progression of breast cancer. However, due to the lack of a relevant genetic model, details of biological consequence of the loss of expression of this gene were unknown. We have constructed and characterized a mammary specific Nmi knock out mouse to elucidate …

Structure/Function Studies Of Yersinia Pestis Metal Transport Systems, Christopher David Radka

All ETDs from UAB

The largest market sector of the global antibiotics industry is on the verge of becoming obsolete because the incidence of β-lactam antibiotic resistance in the clinic continues to rise. Therefore, we are in dire need of new therapeutics to address the increasing threat of antibiotic resistance. Novel targets that could lead to a new drug class are ABC (ATP-binding cassette) importers, which are only found in bacteria. The substrate-binding protein (SBP) components of these transporters present an intriguing subject of study because of their abundance in the cell and potential roles in infection. As a contribution to the scholarship of …

Uncovering The Role Of Common Gamma-Chain Cytokines In Aspergillus Fumigatus Associated Diseases, Kristen Marie Reeder

All ETDs from UAB

Fungal pathogens contribute to a wide range of diseases that are largely dependent on the host’s immune state. In the case of immune suppression, Aspergillus fumigatus is the causative agent of invasive aspergillosis. Furthermore, asthmatics sensitized to fungi such as Aspergillus fumigatus are reported to have more severe asthma. We have previously described a role for IL-22 in mouse models of invasive aspergillosis and fungal asthma. IL-22 was found to play a protective role in invasive aspergillosis, while it was found to play an immunopathogenic role in asthma. Here we describe the cell sources of IL-22 in both models. We …

Striatal Acetylcholine And Dopamine Modulation In A Mouse Model Of Dyt1 Dystonia, Chelsea Zimmerman

All ETDs from UAB

DYT1 dystonia is a neurological movement disorder characterized by sustained muscle contractions causing twisting and repetitive movements or abnormal posture. While many movement disorders are due to loss of neurons, DYT1 dystonia does not present with any gross abnormalities. As of now, the most common form of treatment is anticholinergics. Since anticholinergic are the most effective pharmacotherapy and “paradoxical excitation” has been shown in when quinpirole, D2R agonist, is bath applied on cholinergic interneurons, we began by examining acetylcholine efflux in a DYT1 knock-in mouse model. Our studies provide evidence that at basal levels, there is a hypercholinergic state present …

Streptococcus Pneumoniae In The Heart Subvert The Host Response Through Biofilm-Mediated Resident Macrophage Killing, Anukul T. Shenoy

All ETDs from UAB

During invasive disease, blood circulating Streptococcus pneumoniae are capable of invading the heart and forming bacteria filled cardiac microlesions. These microlesions are devoid of immune cells and disruptive of cardiac functionality. While the mechanisms underlying pneumococcal invasion of the myocardium are well-described, how the heart-invaded pneumococci evade immune detection and clearance is unknown. Since its discovery in 1881, invasive pneumococcal disease has been associated with the presence of extracellular diplococci or short chains in affected tissues. Herein, we show that heart-invaded pneumococci replicate within cellular vesicles and transition into biofilms. Although the host cell permissible for intracellular replication is yet …

Evaluation Of Peripheral And Mucosal T Cell Phenotypes Of Chlamydia-Infected Women, Brian Makori Omwena Ogendi

All ETDs from UAB

Chlamydia trachomatis (CT) infection is the most prevalent bacterial sexually transmitted infection worldwide and women are disproportionately affected due to its reproductive complications. T cell phenotypes present during CT infection have been well characterized in the murine model, but remain to be elucidated in humans. Therefore, studies aimed at increasing our understanding of T cell phenotypes in CT-infected humans were the focus of this dissertation. The first chapter of this dissertation reviews various aspects of chlamydial infection while in the second chapter aims to address the influence of CT infection on T cell phenotypes. We first compared differences in peripheral …

Comparison Of Atypical Bacteriophages That Infect Caulobacter Crescentus, Doreen Nguyen

Theses and Dissertations

Several Caulobacter bacterial genomes have been recently sequenced, and all of the genomes contained one or more clusters of genes with phage origins. This observation indicates that bacteriophages contribute to the Caulobacter gene pool, so in order to understand bacteria genomes we will need to understand phage genomes as well. As part of understanding the phage genomes, we want to isolate novel bacteriophages and study their genomics. This study resulted in the isolation of 12 new phages, including four that differ from the well-studied CbKlike phages. Two of these novel phages are Podoviruses with icosahedral heads and small tails, and …

Environmental Relationships And Predator-Prey Interactions Within The Snapper-Grouper Complex In The Southeastern U.S. Atlantic – Implications For Fisheries Management, Dawn M. Glasgow

Theses and Dissertations

Stock assessments indicate many valuable fish species are declining, or are considered overfished and/or are undergoing overfishing. Fisheries scientists and managers in the southeastern U.S. typically have utilized a single-species approach, and relied on catch per unit effort data derived from fishery-independent surveys to determine indices of abundance for economically important reef fish. However, catchability for these surveys vary for many reasons including environmental and predator-prey relationships. This research was developed to elucidate environmental influences on reef fish assemblages and distribution of reef-associated marine predators, and examine predator-prey dynamics that may influence relative abundance of large predators based on chevron …

Applications Of Multimedia Resources Developed As Part Of The Virtual Cell Animation Collection In Undergraduate Introductory Biology, Eric Edward Goff

Theses and Dissertations

Calls for reform in undergraduate STEM education have arisen from an increased need for well-trained biology graduates in the future scientific workforce. To address this need, many institutions have focused on a pedagogical restructuring of instructional practices to promote deeper conceptual understanding of core biological concepts. This study investigates the implementation of multimedia resources as a possible reliable supplement to undergraduate introductory biology and aims to provide empirical evidence on the instructional best practices of their use. As a central part of this study, we focus on one specific multimedia package, the Virtual Cell Animation Collection, due to its developmental …

Adoptees’ Experiences With Direct-To-Consumer Genetic Testing: Emotions, Satisfaction, And Motivating Factors, Anna Childers

Theses and Dissertations

Public awareness of the role of genetics in disease continues to expand along with the use of direct-to-consumer genetic testing (DTC-GT). One subset of customers taking advantage of the growing DTC-GT market is adoptees. Prior research has shown that adoptees appear to be more motivated than non-adoptees to learn about their genetic disease risk but have similar responses to health-related information acquired through DTC-GT. In this exploratory qualitative study, fourteen adoptees were asked about their motivations for pursuing DTC-GT; satisfaction with results; emotions throughout the process; and interest in meeting with a genetic counselor. Motivations for pursuing DTCGT fell into …

Mast Cells And Lipid Cross-Talk In Skin Inflammation, Piper Alexandra Wedman

Theses and Dissertations

Atopic dermatitis (AD) is an inflammatory skin disease whose pathogenic mechanisms remain unclear. Using a validated human AD-like mouse model, we observed that skin remodeling started at a pre-symptomatic stage of AD that included cellular infiltration of the hypodermis, accompanied with activated/degranulated mast cells (MC). Local MC activation was quantified using a novel method of computer-assisted image analysis we developed and reported. Using a variation of this method, we defined morphometric parameters allowing for quantitation rather than scoring of cellular infiltration. Cell recruitment correlated with MC activation, chemokine production and increased levels of sphingolipid sphingosine-1-phosphate (S1P), produced by sphingosine kinase-1 …

Investigating Novel Methods Of Interaction With Pharmaceutically Relevant Enzymes, Cory T. Reidl

Dissertations

Metalloproteins requiring one or more metal ions for normal function make up 30% of all known proteins, and many critical biological pathways contain at least one metallo-enzyme. Di-nuclear metallo-proteins constitute a large class of these proteins yet we currently lack effective methods of inhibiting these enzymes for the development of new medical therapies, particularly for the discovery of new antibiotics. Our work has focused on developing novel functionalities that selectively interact with di-nuclear catalytic centers, and we are targeting three separate di-zinc-metallo-enzymes that are unique to bacteria and play key roles in their growth and development. These enzymes are DapE, …

Elevated Hydrostatic Pressure Selectively Enhances Matricellular Gene Expression In Human Trabecular Meshwork Cells, Jonathan Lautz

Dissertations

Primary open-angle glaucoma (POAG) is a progressive optic neuropathy

characterized by loss of peripheral vision secondarily associated with elevated intraocular pressure (IOP). Transforming growth factor (TGF)-β2 is markedly elevated in the aqueous humor (AH) of patients with POAG. It has been previously shown that TGF-β2 increases IOP, in part, by inducing expression and release of endothelin-1 (ET-1) and connective tissue growth factor (CTGF) within the trabecular meshwork (TM). ET-1 and CTGF may ultimately lead to increases in IOP by enhancing TM cell contractility and extracellular matrix (ECM) deposition. Here we confirm that TGF-β2 enhances mRNA content of ET-1, CTGF, and …

Protein Profiling Analysis Of Platelets In Hypercoagulable State Of Β-Thalassemia/Hbe Patients, Puangpaka Chanpeng

Chulalongkorn University Theses and Dissertations (Chula ETD)

β-thalassemia/HbE is an inherited hemolytic anemia caused by defect in β-globin synthesis resulting in accumulation of excess α-globin chains in red blood cells. A hypercogulable state leading to high risk of thromboembolic event is one of the most common complications observed in this disease, particularly in patients with splenectomy. Previous studies suggested that increased platelet activation and coagulation factors in β-thalassemia/HbE intermediate patients promote the hypercoagulable state. However, the hypercoagulable state as well as the molecular mechanism regarding this pathogenesis in β-thalassemia/HbE is not yet well understood. This study aimed to identify proteins related to platelet activation and to hypercoagulable …

Comparison Between Digital Image Analysis And Visual Assessment Of Immunohistochemical Her2 Expression In Breast Cancer, Morten Ragn Jakobsen

Chulalongkorn University Theses and Dissertations (Chula ETD)

Background: Assessment of HER2 status is considered standard of care in the histopathologic workup of breast cancer and conveys prognostic and predictive information used to guide treatment decisions. The assessment is often carried out in a two-step approach where immunohistochemical expression of HER2 protein is first evaluated by conventional microscopy and equivocal cases are further analyzed by in-situ hybridization techniques to assess gene amplification status. Methods: In this study we compared conventional manual assessment of immunohistochemical HER2 expression with digital image analysis (DIA) and consensus manual assessment by a panel of three pathologists. From our archive we retrieved sections of …

Pretreatment Prognostic Factors To Predict Survival Outcome In Advanced Non-Small Cell Lung Cancer With First Line Treatment In Thailand: A Retrospective Cohort Study, Sureerat Jaruhathai

Chulalongkorn University Theses and Dissertations (Chula ETD)

Background: Tumor genetic information and biologic markers are often used as prognostic factors. However, these have limited applicable in real daily practice due to their high cost . Objective: To evaluate which inexpensive, convenient simple prognostic factors are associated with survival outcome with first line treatment among patients with advanced non-small cell lung cancer (NSCLC). Methods: Retrospectively reviewed the charts of patients diagnosed with having advanced NSCLC with no previous treatment seen at King Chulalongkorn Memorial Hospital and Police General Hospital during 1 January 2008-31 Dec 2013. Results: A total of 301 patients were included in the study; 68.1% males. …

Involvement Of Notch Signaling In Interleukin-4-Stimulated Human Macrophages, Naunpun Sangphech

Chulalongkorn University Theses and Dissertations (Chula ETD)

Macrophages respond to various stimuli, resulting in distinct effector phenotypes that range from LPS-stimulated proinflammatory to IL-4-activated pro-healing phenotype. The regulatory mechanism of this polarization is not fully understood. Notch signaling pathway is a conserved signaling pathway involved in polarization of macrophages, especially proinflammatory macrophages. However, the role of Notch signaling in IL-4 stimulated macrophages (M(IL-4)) is not well defined. PPARgamma is a signature nuclear receptor in M(IL-4) that function mainly in regulating lipid metabolism. There are many reports that revealed the interaction of Notch signaling and PPARgamma in various cells but not in macrophages. In this study, the interaction …

Development Of Gold Nanoparticle-Based Lateral Flow Immunoassay For Detection Of Pathogenic Leptospiral Antigen, Prayoon Lae-Ngee

Chulalongkorn University Theses and Dissertations (Chula ETD)

Leptospirosis, caused by pathogenic Leptospira spp., is a global re-emerging zoonosis and endemic disease in tropical and sub-tropical countries including Thailand. Point-of-care testing (POCT) for rapid diagnosis of leptospirosis is needed for prompt and appropriate treatment particularly in resource-poor settings. Lateral flow immunoassay (LFA)-based POCT devices are rapid, user friendly, cheap, have one-step analysis, and need no sophisticated equipment. LipL32 is a common target for diagnostic tests of leptospirosis because it is the major outer membrane protein (OMP) that is specific and highly conserved among pathogenic Leptospira. This study aimed to develop the gold nanoparticle (AuNP)-based LFA for detection of …

Characterization Of T Cell Secreted Cytokines And Cytotoxic Mediators Profiling In Knee Osteoarthritis Patients, Thitiya Sae-Jung

Chulalongkorn University Theses and Dissertations (Chula ETD)

Osteoarthritis (OA) is a chronic disease associated with morphological joint changes that results in the breakdown of joint cartilage and underlying bone with erosive inflammation. Factors of OA are intrinsic factor and extrinsic factor. The pathogenesis of OA relates with immune responses and immune cells, T cells, B cells, and macrophages etc., T cells both CD4+ and CD8+ T cell are immune cells that have been suggested to play a role in OA. However, we found many finding in CD8+ T cells such as a high proportion of CD8+ T cell was observed in tissue of knee joint. In addition, …

Immune Response In Pythiosis Patients Treated With Pythium Insidiosum Antigen (Pia), Navaporn Worasilchai

Chulalongkorn University Theses and Dissertations (Chula ETD)

Immunotherapy using Pythium insidiosum antigen (PIA) is one of the combination treatment for P. insidiosum infected patients along with amputation and certain antifungal agents. The protocol of first prime (day0) after diagnosis and six booster doses (at 0.5, 1, 1.5, 3, 6, 12 months; mo.) along a year period were performed based on previous study. The switching of T-helper2 (Th2) to Th1 after immunotherapy along with the clinical improvement in equine and rabbit models were revealed whereas studies in human are very limited. To investigate the immune response in PIA treated pythiosis patients, cytokine profile (CMI response), P. insidiosum specific …

The Antiviral Activity Of Phenolic Lipids Group Against Dengue Virus, Parichat Kanyaboon

Chulalongkorn University Theses and Dissertations (Chula ETD)

Dengue virus causes a global burden that specific chemotherapy has not been established in a clinical setting. A previous report suggested that anacardic acid saturated (C15:0), a member of phenolic lipids, inhibited hepatitis C virus infection. Here, we explored structure activity relationship of phenolic lipids homologues with anti-DENV properties in the cell-based system. Our results indicated that Cardol triene (C15:3) expressed DENV1-4 inhibition, but not to Zika virus (SV0010/15) with the EC50s of 5.35 ± 1.16 µM, 7.13 ± 0.72 µM, 8.98 ± 1.77 µM, and 8.21 ± 0.34 µM, respectively. Also, the compound showed good CC50s to Vero cells …