Cell and Developmental Biology Commons^™

Crippling Rapid Evolution Of Metastasis And Drug Resistance In A549 Non-Small Cell Lung Cancer Cells With The Clinically Relevant Hsp90 Inhibitor Auy922, Nickolas Anthony Bacon

Theses, Dissertations and Capstones

The ability for species to evolve new features in response to changing circumstances in order to survive and propagate is a ubiquitous observation on both the macroscopic and microscopic levels of living systems. It should be no surprise, then, that diseases such as cancer utilize their own forms of adaptation to perpetuate themselves when exposed to external threats. Indeed, concepts drawn from Darwinian evolution are now widely accepted to help explain certain aspects of carcinogenesis and malignant progression, the sum of which have come to be known as the theory of tumor evolution. Since metastasis and drug resistance are features …

Na/K-Atpase Alphα1 Regulates Adipogenesis Via Its Conserved Caveolin Binding Motif, Minqi Huang

Theses, Dissertations and Capstones

The Na/K-ATPase (NKA) was identified in 1957 by Dr. Jens C. Skou. It belongs to the P-type ATPase family, which can actively transport ions across cell membranes by using the energy from adenosine triphosphate (ATP) hydrolysis. During the second half of the 20th century, the molecular mechanism of the NKA catalytic cycle was clarified, and the isoform diversity of NKA in different species and organs was identified. The active ion transport through NKA generates cell membrane ion gradients and the electric potential. Hence, the enzymatic function of NKA is critical for cell viability as well as multiple physiological processes including …

Potential Counter Regulatory Effects Of A Gut Microbiota Metabolite In Alleviating Down-Regulation Krüppel-Like Factor 4 In Intestinal Inflammation, Ylva Forslund

Theses, Dissertations and Capstones

Inflammatory bowel disease (IBD) is a medical condition characterized by chronic inflammation of the intestinal epithelium. Krüppel-like factor 4 (KLF4), a zinc finger transcription factor, is vital for maintaining intestinal epithelial homeostasis. KLF4 promotes differentiation of goblet cells that generate the protective mucus layer. Reduced goblet cell number and defective mucus layer are associated with IBD. Shortchain fatty acids (SCFA) are known to play an important role in the maintenance of a strong and healthy intestinal epithelial layer and also in goblet cell differentiation. However, whether the positive effects of SCFAs on goblet cells are mediated, at least partly, via …

Targeting Epigenetic Mechanisms In Endometriosis, Sarah Elizabeth Brunty

Theses, Dissertations and Capstones

Endometriosis is a complex and elusive gynecological disease in which the inner lining of the uterus grows in locations outside of the uterus and forms lesions. It is known to affect 1 in 9 women of reproductive age worldwide. Symptoms of endometriosis include severe pain, heavy periods, and infertility. While multiple theories of origin exist, none fully encompass all aspects of the disease, although all theories agree that this is an inflammation-driven disease. Due to this, many researchers are turning towards epigenetics to explain the initiation and progression of endometriosis. However, what is causing these epigenetic changes is still a …

Isolation And Identification Of Hsc70 Conjugates In Raw264.7 Murine Macrophage-Like Cells, Michael Arland Parsons

Theses, Dissertations and Capstones

IL12R- β2 is a Type I cytokine receptor and contains the WXXW (WSNWS) sequence that often predicates the post-translational addition of mannose to a tryptophan residue via a carbon-carbon bond. This study will stimulate expression of IL12R- β2 in RAW 264.7 Macrophage-Like Cells by shutting down the extracellular signal kinase (ERK) pathway and introducing inflammatory agents lipopolysaccharide and CpG DNA in order to collect a concentrated sample of IL12R- β2. These samples will be analyzed for the presence of C-mannosyltryptophan residue

Influence Of Developmental Stage, Habitat, And Captivity On Thecutaneous Bacterial Communities Of Eastern Hellbenders (Cryptobranchus Alleganiensis Alleganiensis) In West Virginia, Rachel Fern Arrick

Theses, Dissertations and Capstones

Microbes inhabit virtually all surfaces of multicellular animal hosts, with microbial cells outnumbering the hosts’ own cells 10:1. Symbiont microbes, collectively referred to as the microbiome, can have profound impacts on the metabolism, development, behavior, and disease resistance of their multicellular hosts. Because the community structure of symbiont bacteria can influence host health, the characterization of amphibian microbiomes is becoming an increasingly important tool for future conservation in the face of global amphibian declines. Eastern hellbenders are good candidates for a microbiome study because they have seen substantial declines in recent decades and learning more about the environmental and physiological …

Uncovering New Roles For The Aryl Hydrocarbon Receptor (Ahr) In Breast Cancer, Justin Kirk Tomblin

Theses, Dissertations and Capstones

The aryl hydrocarbon receptor (AHR) is a ligand-activated transcription factor that is responsive to many exogenous and endogenous ligands. AHR is of particular interest in cancer, and has been shown to play roles in tumor progression. As such, it has received growing attention as a possible chemotherapeutic target. Obesity increases the risk of breast cancer, complicates treatment of breast cancer, and stimulates the growth of larger, more aggressive mammary tumors. Many breast tumors in obese women are estrogen receptor (ER)-positive and, while targeting hormone receptors like ER is beneficial, many obese women see a recurrence of their cancer after standard …

Adult Neural Stem Cell Differentiation And Signaling Is Disrupted By Low-Level Silver Nanoparticle Exposure In Vitro, Robert Jefferson Cooper

Theses, Dissertations and Capstones

Silver nanoparticles (AgNP) are an emerging environmental contaminant with unique chemical and physical properties. They are utilized in products like medical dressings and children’s toys for their antimicrobial action. Though AgNP’s bioaccumulative nature and high-level toxicity are established, low-level effects from chronic exposure to AgNP-containing products and environmental AgNP remain unclear. This study uses adult neural stem cells, a model for neural cell function and neurodevelopment, to assess changes in cell differentiation and behavior following low-level AgNP exposure. In vitro, low-level AgNP produced dose- and timedependent formation of co-localized f-actin inclusions and β-catenin puncta. Neurite extension and arborization were also …

3,5-Dichloroaniline: Biotransformation And Mechanistic Aspects Of Nephrotoxicity In Vitro, Christopher Robert Racine

Theses, Dissertations and Capstones

Chlorinated anilines have been used as important chemical intermediates in the production of a wide variety of pharmaceutical, industrial, and agricultural products. Exposure to chloroanilines can arise in both occupational and environmental settings and can lead to toxicity to multiple organs, including the kidney. Previous studies have established that of the mono- and dichlorinated anilines, 3,5-dichloroaniline (3,5-DCA) possesses the most nephrotoxic potential both in vitro and in vivo. However, little is known concerning the role of renal metabolism in nephrotoxicity. The studies presented in the following dissertation are the first to explore the toxicity, metabolism, and bioactivation of 3,5-DCA in …

Epigenetic Role Of Ptip In Mouse Spermatogenesis, Chengjing Liu

Theses, Dissertations and Capstones

In mammals, spermatogenesis is a biological process inside the testis to produce spermatozoa from spermatogonia. This process is governed by both genetic and epigenetic mechanisms and thus is a powerful system for epigenetic research. Methylation of histone 3 lysine 4 (H3K4) is an epigenetic mark, which has been found to be dynamically modulated in mouse male germ cells during spermatogenesis. Pax2 Transactivation domain Interaction Protein (PTIP) has been recently identified as part of a H3K4 methyltransferase complex. In this study, I hypothesize that PTIP is an essential epigenetic regulator in mouse spermatogenesis. To test this hypothesis, I first established a …

Cerium Oxide Nanoparticles Attenuate Polymicrobial Sepsis Induced Splenic Damage In Male Sprague Dawley Rats, Venkata Vinay Kumar Bandarupalli

Theses, Dissertations and Capstones

Sepsis is a serious life threatening medical emergency which, if not treated properly, oftentimes results in organ failure and death. Current sepsis treatment protocols are largely centered on the use of antibiotics and supportive care. Recent studies have suggested that antibiotics fail to be effective for sepsis treatment when administered during hypo-dynamic phase of sepsis that is usually characterized by the presence of a cytokine storm. As such, there is an urgent need to develop novel therapeutic drugs that target the inflammatory cytokines that are secreted as a result of increased reactive oxygen species. Cerium oxide nanoparticles (CeO2) have been …

Chmp1 Negatively Regulates Epidermal Growth Factor Signaling In The Drosophila Wing, Meagan Elisabeth Valentine

Theses, Dissertations and Capstones

A critical step in cellular signaling through transmembrane receptors is the down-regulation of activated receptors through the multivesicular body (MVB) pathway to the lysosome. MVB generation is mediated by the highly conserved ESCRT (0, I, II, and III) protein complexes. Though the ESCRT-III complex provides the core function of the ESCRT machinery, it is the least characterized of the ESCRT complexes. The Chmp1 protein is an ESCRT-III component and a putative tumor suppressor that has been linked to pancreatic and renal cancers in humans. However, published data on Chmp1 activity are conflicting and its role during tissue development is not …

Gene Transfer In A Newly Established Spontaneous Feline Fibrosarcoma Cell Line, Rounak Nande

Theses, Dissertations and Capstones

Fibrosarcoma is a deadly disease in cats and is most often located at classical vaccine injections sites. More rare forms of spontaneous non-vaccination site (NSV) fibrosarcomas have been described and have been found associated to genetic alterations.

Purpose of this study was to compare the efficacy of adenoviral gene therapy in NVS fibrosarcoma. The NVS fibrosarcoma cell line (Cocca-6A) was isolated from a spontaneous fibrosarcoma that occurred in a domestic calico cat. Cocca-6A fibrosarcoma cells were morphologically and cytogenetically characterized. The feline cells were karyotyped and their chromosome number was counted using a Giemsa staining. Giemsa block staining of metaphase …

Interaction Of Pyrimoidiazolium Ions With Calf Thymus Dna, Stormy Rene Gibson

Theses, Dissertations and Capstones

A new class of fluorescent dyes, pyridoimidiazolium cations, is being considered for their possible use in the biochemical field because of their interactions with DNA. Binding of low molecular weight fluorophores has been proven to cause a wide variety of biological responses in DNA. The dyes used here have been synthesized using α-diimine type compounds. The reaction used has the capability to produce many fluorescent cations. Binding constants of these compounds in acetonitrile with calf thymus DNA have been determined by measuring emission intensity. Emission intensity showed the degree to which binding is occurring. The experimental results show that binding …

Omega-3 Fatty Acids As Therapeutic Options For The Treatment Of B-Cell Chronic Lymphocytic Leukemia, Johannes Francois Fahrmann

Theses, Dissertations and Capstones

B-cell chronic lymphocytic leukemia (CLL) is the most common form of adult leukemia in the western world. CLL is often diagnosed in the asymptomatic (early-stage) stages. However, approximately 50% of these patients will progress to advanced, symptomatic disease and require therapy. Current treatment options are limited due to progressive drug resistance and severe drug-induced toxicities which are often too toxic for the elderly or those with co-morbidities. Therefore, a non-toxic therapeutic intervention that could slow the progression of asymptomatic CLL to symptomatic CLL or enhance the effects of actively used chemo-therapeutic drugs in patients who require therapy would be clinically …

Bursicon, Metamorphosis And Development Of Resilin In The Fruit Fly, David Paul Anthony Neff

Theses, Dissertations and Capstones

Hovering flight is the primary form of locomotion in the numerous and diverse forms of winged insects. A specialized elastic protein, Resilin, contributes to flight mechanics in most described insect orders. Although many studies have inferred functions of Resilin in insect flight, we report the first direct evidence that Resilin is a necessary component in flight mechanics of the fruit fly. We have successfully knocked down Resilin synthesis with RNAi and have generated a new resilin deletion mutant Df(2R)anhedral. By disturbing the formation of the Resilin rich tergopleural tendon, we have shown that Resilin functions in wing control. We also …

Mechanisms Of Yttrium Oxide Toxicity In Hek293 Cells, Sravanthi Bodapati

Theses, Dissertations and Capstones

As a non-metal oxide, yttrium oxide (Y2O3) nanoparticles have numerous applications in chemical synthesis, mechanical polishing and as additives to drugs, cosmetics, varnishes and food. Recent data have suggested that these particles are capable of inducing oxidative stress and cytotoxicity in human endothelial cell lines. To examine the potential mechanisms of yttrium oxide toxicity, human embryonic kidney (HEK293) cells were exposed to 1, 5, 10, 50 and 100 μM of Y2O3 nanoparticles for 12, 24, 36 or 48 hr. We hypothesized that exposure of HEK293 kidney cells to Y2O3 nanoparticles would be associated with increased evidence of intracellular oxidative stress …

The Role Of Chmp1 In Drosophila Melanogaster, Meagan E. Valentine

Theses, Dissertations and Capstones

Chmp1A has recently been linked to pancreatic cancer, a leading cause of cancer death in humans. Pancreatic tumors have lowered Chmp1A expression, and it has been described as a tumor suppressor. Chmp1A is also a member of ESCRT III (Endosomal Sorting Complex Required for Transport), a conserved protein complex involved in the degradation and recycling of activated transmembrane receptors. There is a single Chmp1 protein in Drosophila that is homologous to vertebrate Chmp1A; however, Chmp1 hasn’t been studied in Drosophila. The objective of this study was to characterize Chmp1 in Drosophila using gene knockdown and over-expression. We used an …

Characterization And Differentiation Of Peripheral Blood Derived Multipotent Adult Progenitor Cells, Hari Satya Shankar Addagarla

Theses, Dissertations and Capstones

Stem cells are populations of undifferentiated cells that are found in most tissues and act as precursors for regeneration and maintenance. In the future, they could provide promising therapies for diseases which are to date incurable. Our lab developed a novel cell line from the peripheral blood of adult transgenic green fluorescent protein swine and designated them as Peripheral Blood Derived Multipotent Adult Progenitor Cells (PBD-MAPCs). In this study we characterized the mRNA and protein expression profiles of PBD-MAPCs before and after neural differentiation and investigated the potential of PBD-MAPCs to differentiate into myocardial or neural lineages in vitro. We …

Loss Of Ski Expression In Testicular Cancer Leads To An Enhanced Invasive Phenotype Through Both Bmp-Dependent And Bmp-Independent Pathways, Amy N. Nash

Theses, Dissertations and Capstones

The proto-oncogene SKI is a transcription factor and a co-repressor of the TGFβ superfamily, including TGF

β and BMP. However, additional data suggests that SKI may function as a tumor suppressor in some cell types. The TGFβ superfamily has been implicated in cancer progression and germ cell migration. Testicular cancer afflicts men during their peak reproductive years and is the most common cancer among men of this age group. Cisplatin-based chemotherapy is the standard treatment for testicular cancer. This treatment can lead to undesirable side effects, including infertility. We have shown that SKI expression is decreased in testicular germ cell …

Enumeration Of Endothelial Progenitor Cells In Hind Limb Suspended Rats And The Mechanics Of Endothelial Wound Healing, Jarrod Matthew Pennington

Theses, Dissertations and Capstones

The endothelium repairs itself through two methods. One is by the activity of circulating endothelial progenitor cells (EPCs). EPCs are immature endothelial cells that circulate the bloodstream that are capable of proliferation and differentiation into mature endothelium. It is thought that EPCs contribute to the repair and replacement of damaged endothelial cells in the process of reendothelialization. Physical inactivity and bed rest are known to be deleterious to the endothelium. It is possible that this inactivity is also deleterious to the number of viable EPCs, which would reduce the ability of the endothelium to repair itself. We used hind limb …

Effects Of Aging On Regulators Of Muscle Apoptosis In The Female F344bn Rat, Murali K. Gadde

Theses, Dissertations and Capstones

Age-related muscle atrophy is a consequence of normal aging characterized by decreases in muscle mass and strength. The mechanism(s) underlying the loss of muscle mass with increasing age is not fully understood, however recent data has suggested that muscle cell apoptosis may be involved. Here we investigate how aging affects the regulation of muscle apoptosis in the extensor digitorum longus (EDL) and soleus muscles of young (6-month), aged (26-month), and very aged (30-month) female Fischer 344/NNiaHSD X Brown Norway / BiNia (F344BN) rats. EDL and soleus muscle mass/body weight ratios were lower in aged animals but not different between 26- …

Comparing Digit Morphology Of An Arboreal Salamander With Potential Competitors, Eric Hugh Diefenbacher

Theses, Dissertations and Capstones

Green Salamanders (Aneides aeneus) are the only salamanders in West Virginia to exhibit an arboreal lifestyle. The focus of this study was to determine the cellular anatomy of the distal digit structures and how these structures may influence climbing ability. Comparative histology, comparative morphometrics, and comparative osteology of Green Salamanders, Cumberland Plateau Salamanders (Plethodon kentucki), and Slimy Salamanders (Plethodon glutinosus), were also used to determine if Cumberland Plateau Salamanders had the potential to compete with Green Salamanders for arboreal habitats. Histologically, Cumberland Plateau Salamanders had cell layer thickening similar to that of Green Salamanders. Morphometrically, Green Salamanders and Cumberland Plateau …

Distribution And Conservation Genetics Of The Cow Knob Salamander, Plethodon Punctatus Highton (Caudata: Plethodontidae), Matthew R. Graham

Theses, Dissertations and Capstones

Being lungless, plethodontid salamanders respire through their skin and are especially sensitive to environmental disturbances. Habitat fragmentation, low abundance, extreme habitat requirements, and a narrow distribution of less than 70 miles in length, makes one such salamander, Plethodon punctatus, a species of concern (S1) in West Virginia. To better understand this sensitive species, day and night survey hikes were conducted through ideal habitat and coordinate data as well as tail tips (10 to 20 mm in length) were collected. DNA was extracted from the tail tips and polymerase chain reaction (PCR) was used to amplify mitochondrial 16S rRNA gene fragments. …

The Expression And Function Of Ppar And Hif-1 In Human Melanoma, Caroline Mills

Theses, Dissertations and Capstones

The first part of my dissertation focuses on the expression and function of PPARs in human melanoma. I found that the A375 cells were significantly growth inhibited in response to _PGJ2 and troglitazone treatment. HEMn-LP showed significant growth inhibition in response to troglitazone. I found that PPARγ and PPARδ mRNA is present in both the SK-Mel 28 and A375 cells. The relative level of PPARα mRNA expression is highest in SK-Mel 28 cells, ~3 fold higher relative to both the normal human melanocytes and A375 cells. PPARγ protein was ~50% higher in both SK-Mel 28 and A375 cells relative to …

New Microanatomical Characters For Scorpion Taxonomy (Arachnida: Scorpiones), Michael Brewer

Theses, Dissertations and Capstones

Morphological characters used in scorpion systematics are limited to a few reliable structures. The most recent high-level, comprehensive revision of scorpion phylogeny, 38 of the 105 morphological characters were based on trichobothria – socketed mechanosensory seta (Soleglad & Fet, 2003). Other microstructures such as granules, setae, spines, etc. have been found in scorpions but their use for taxonomy was limited. In an effort to better understand the complex relationships among these ancient but similar looking animals, we attempted to first locate new microstructures and then assess their reliability and variability thus determining the levels at which they are diagnostic. The …

Arsenic Toxicity In Plhc-1 Cell Line And The Distribution Of Arsenic In Central Appalachia, Yeong Nam Jeong

Theses, Dissertations and Capstones

Our ecosystem has geobiophysical relationships. Environmental pollution can lead to immediate consequences like organism death or lack of reproduction. It can also lead to long-range consequences through mutations and DNA damage. Fish can be affected very directly by environmental contamination in water but the study of fish is not performed well. We are developing a quantitative assay to assess DNA damage in vitro due to heavy metal contamination using a tissue culture line of Poeciliopsis lucida, the desert pupfish, which is used for other toxicology assays. To detect DNA damage, Single Cell Gel Electrophoresis (Comet Assay) is very useful. …

Chmp1 Is Implicated In The Development Of Pancreatic Tumor Via The Retinoic Acid Signaling Pathway, Juliana Adedayo Akinsete

Theses, Dissertations and Capstones

In the present study, we investigated the involvement of Chmp1 (Chromatin ModifyingProtein 1/Charged Multivesicular body Protein 1) in the development of mousepancreatic acinar tumor cell line. CRL 2151 cell line was transfected with Chmp1/CS2vector to compare growth, morphology and expression of Chmp1, p53 and pp53 (ser37) with control-transfected cells. CRL 2151 cells were treated with all-trans retinoicacid (ATRA) to compare growth, morphology and expression of Chmp1 and p53 withcontrol-treated cells. Strabismus was used as control. Results showed inhibition ofgrowth but no morphological change in transfected cells. Western blot analysis showedthat Chmp1 transfection upregulated the expression of p53, pp53 and Stbm …

Characterization Of The Dna-Binding Properties Of Silent Information Regulator 3 Protein, Cotteka Nichisha Johnson

Theses, Dissertations and Capstones

The eukaryotic genome is organized into distinct domains with discrete regulatory potential, influenced by the variation in local chromatin structure. An example of chromatin-mediated consequences on gene expression is mating type determination of yeast Saccharomyces cerevisiae, which depends on silencing of specific gene cassettes. Silencing in yeast involves key proteins, amongst which the Silent Information Regulator (Sir) proteins are essential. Sir1p, Sir2p, and Sir4p are recruited to nucleate silencing events, while Sir3p serves as the pivotal factor in spreading the silent state. This investigation characterizes the DNA binding properties of Sir3p using DNA fragments with varying degrees of inherent flexibility, …

Analysis Of Genotypic And Phenotypic Differences In Desmognathus Quadramaculatus Across The Southern Appalachians, Deborah Susan Merritt

Theses, Dissertations and Capstones

Populations of Desmognathus quadramaculatus were compared for genotypic and phenotypic differences across 5 states in the Southern Appalachians. Series consisting of 10 salamanders each were collected from West Virginia, Virginia, Tennessee, North Carolina and Georgia. Salamanders were assessed for genotypic differences by using allozyme (proteins with polymorphic loci) electrophoresis staining for 13 loci of the muscle, liver and stomach tissue. Nei’s genetic distance was used to score genetic variation between populations. Phenotypic differences were assessed by comparing 10 external morphological measurements between populations. Mahalanobis’ distance was used to score morphological differences between populations. Both genetic and morphological differences were correlated …