Cell and Developmental Biology Commons^™

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 …

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 …

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. …

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 …

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. …

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, …

Aging Affects Stretch-Induced P70s6k And 4e-Bp1 Phosphorylation In Fast- And Slow-Twitch Muscle, Sreevani Uddemarri

Theses, Dissertations and Capstones

In the present investigation we compare the expression, basal activation and the ability of muscle stretch to activate the p70^S6k pathway in the fast-twitch extensor digitorum longus (EDL) and slow-twitch soleus of adult (6 mo. old), aged (30 mo. old) and very aged (36 mo. old) Fischer 344 x Brown Norway rats. Immunoblotting demonstrated that the tissue content of mTOR, p70^S6k, 4E-BP1 and GSK-3β decreased in the EDL and soleus in aged rats, while SHP-2 increased in the EDL and decreased in the soleus when compared to adult rats. Basal phosphorylation of 4E-BP1 increased in both the …

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 …

Semi-Biosynthesis Of Dna Nanostructures, Aoune Barhoumi

Theses, Dissertations and Capstones

Nanotechnology refers to all technologies aiming to build objects, make measurements, and carry out processes on the nanometer length scale. In particular molecular nanotechnology exemplifies the so-called "bottom up" approach, which is briefly defined as the ability to build useful nanostructures with molecular precision, such as molecular machinery. Such capability for controlling matter at the molecular scale has always been the dream of scientists.

All living things are nanofoundries. Billions of years ago, nature perfectly provided all living things with the most accurate biological nanotechnology systems. Cellular internal dynamics, communicative resonance in protein conformational states, viruses as microreplicators, nanoscale life …

Developing Methods For The Rapid Molecular Assessment Of Aquatic Microbial Communities, Traci D. Hudson

Theses, Dissertations and Capstones

Molecular investigations of microbial community structure and dynamics involve costly and time-consuming methods. This approach is limiting when rapid assessment and detection of microbial organisms are needed. In aquatic environments, especially freshwater environments which may be used as a water source, rapid detection of pathogenic microbes is essential. Likewise, monitoring for the presence or absence of various functional genes can be used to indicate the type of microbial community present in the environment of interest. Therefore, the aim of this study was to develop and optimize methods necessary for the rapid assessment of freshwater microbial community structure and dynamics. Primers …

A Mechanistic Study Of Myoglobin Nephrotoxicity, Jennifer L. Minigh

Theses, Dissertations and Capstones

Myoglobin is an endogenous protein that can become nephrotoxic under certain conditions such as crush injuries, drug overdose, and seizures where prolonged contraction of muscle leads to cell death and leakage of myoglobin. The mechanism of myoglobin-induced nephrotoxicity is not fully understood. The purpose of this study was to characterize the sequence and mechanistic events associated with the in vitro toxicity of myoglobin in renal cortical slices. Renal tissue was isolated from Fischer 344 rats. Slices of renal cortex were prepared by freehand. These slices were then incubated for 60-180 minutes with myoglobin (0-12 mg/mL) pretreated with 4 mM ascorbic …

Pkcα Translocation And Actin Remodeling In Contracting A7r5 Smooth Muscle Cells, Chenwei Li

Theses, Dissertations and Capstones

Recent research indicates that protein kinase C (PKC) plays an important role in smooth muscle contraction. Because PKC activation and specificity of substrate phosphorylation is believed to be associated with the relocalization of the enzyme to specific cell sites, we first investigated the subcellular translocation of PKCα in A7r5 smooth muscle cells by confocal microscopy through use of standard immunohistologic staining and PKCα - enhanced green fluorescent protein (PKCα -EGFP) fusion protein expression. PKCα was diffusely distributed throughout the cytosol in the unstimulated A7r5 cell. Upon stimulation with phorbol 12, 13 dibutyrate (PDBu), PKCα was translocated primarily to either the …

Actin And Myosin Remodeling In The A7r5 Smooth Muscle Cell, Michael E. Fultz

Theses, Dissertations and Capstones

Active remodeling of the cytoskeleton has been proposed to contribute to the low energy cost and maintenance of the sustained contraction in smooth muscle. Using confocal microscopy and standard immunohistochemical techniques, direct observation of actin remodeling was studied in the contracting A7r5 cell in response to the diacylglycerol (DAG) analog phorbol 12, 13 dibutyrate (PDBu). -actin was shown to exist in the resting cell as parallel stress cables that extend across the cell. Stimulation by PDBu resulted in a sustained contraction that occurred in approximately eighty-five percent of the A7r5 cells. The initial contraction was not uniform, but primarily occurred …

Analysis Of E. Coli Dna From Selected Source(S) Using Pulsed-Field Gel Electrophoresis, Sheema Doshi

Theses, Dissertations and Capstones

The presence of fecal coliforms in surface waters is a public health issue throughout the United States and Canada. The presence of Escherichia coli (E. coli) indicates recent fecal contamination from either animal or human sources. To solve this growing problem one must identify the source(s) of such contamination by creating a database of DNA profiles of£. coli isolates. Hence, tracking the source(s) of E. coli can help link water contamination to animal source(s) of contamination. In this study, pulsed field gel electrophoresis (PFGE) was used to analyze DNA from E. coli isolates from known animals. For PFGE, the bacterial …

Genomic Diversity Of Pseudomonas Aeruginosa In Cystic Fibrosis, Nathaniel Edwards Head

Theses, Dissertations and Capstones

Pseudomonas aeruginosa is a common environmental microorganism. However, it has the genetic capacity to cause diseases in patients with compromised host defense systems. Cystic fibrosis (CF) is one of the major hereditary diseases among Caucasian populations. CF patients are born with a defective chloride channel that is responsible for maintaining fluid and electrolyte balance across the lumen of the lung. This imbalance leads to the production of an abnormal dehydrated viscous mucous, prohibiting the patient from normally clearing the respiratory airway. Chronic pulmonary infections with P. aeruginosa are the major causes of high morbidity and mortality in CF. Two of …

Factors Influencing Spatial Variability In Soil Nitrogen (N) Dynamics In N-Treated And Untreated Watersheds Of The Fernow Experimental Forest, West Virginia, Nikki Lenore Lyttle

Theses, Dissertations and Capstones

The central Appalachian region of the United States receives some of the highest inputs of nitrogen (N) due to acidic deposition in the nation. It is believed that these high could levels contribute to a decline in forest soils within the next 50 to 70 yrs. This study examines factors that influence spatial variability in N-treated and untreated watersheds of the Fernow Experimental Forest, Parsons, West Virginia. Within each of the two watersheds [WS4 untreated control, > 100 yr.; WS3 N-treated, acidified, clear cut, ~ 31 yr.], two 0.04 ha plots, one high N and one low N, were selected for …

Characterization And Regulation Of Skn-Directed Autoimmune Skin Lesions, Pamela Jane Staton

Theses, Dissertations and Capstones

In the Skn model of autoimmunity, pathogenic T cells sensitized to Skn epidermal antigens (Skn-immune spleen cells or SIS cells) elicited disease in the form of skin lesions when adoptively transferred into shaved immunoincompetent syngeneic adult recipients (‘lesion-forming animals’) while concomitant injection of immunoregulatory normal spleen cells at twice the concentration (2 x normal spleen cells or 2XNS cells) of SIS cells significantly lessened lesion development (‘lesion-controlling animals’) by Day 7 post cell transfer. In a time-course RT-PCR cytokine analysis of skin from lesion-forming and lesion-controlling animals, IL-7 mRNA was significantly elevated in lesion-controlling skin on Day 7 while no …

The Life History Of The Cave Salamander, Eurycea Lucifuga Rafinesque, In West Virginia, Andrew James Longenecker

Theses, Dissertations and Capstones

In West Virginia, as throughout its range, there is limited information about the life history of the cave salamander (Eurycea lucifuga). The purpose of this study was to describe the natural history of this species in West Virginia. The objectives of this study were to determine habitat selection of the cave salamander, to ascertain its environmental characteristics, and to study its reproductive and non-reproductive biology. Three study caves were located in Greenbrier County, West Virginia. All sites were examined at least once each month from May 1999 through April 2000. Caves were divided into three zones: entrance, twilight, …

Ecology And Sympatric Relations Of Crevice Salamanders In Randolph County, West Virginia, Jayme Linn Waldron

Theses, Dissertations and Capstones

By implementing a mark-recapture study, I investigated the ecology of three species of sympatric plethodontid salamanders (Aneides aeneus, Desmognathus ochrophaeus, and Plethodon glutinosus) on rock outcrops in the Westvaco Wildlife and Ecosystem Research Forest, Randolph County, West Virginia. By examining activity patterns, vertical stratification, and habitat preference for A. aeneus, D. ochrophaeus, and P. glutinosus, I attempted to offer some insight into how these sympatric species avoid competition on small, isolated rock outcrops. Twenty-one surveys, both nocturnal and diurnal, were performed between 11 May and 17 October, 1999. During this time, 89 salamanders, encompassing five species ( …

Whole Animal And Cellular Strategies In The Regulation Of Bone Mass In The Rat, Weidong Geng

Theses, Dissertations and Capstones

In order to determine the strategies in regulation of bone mass in the rat and better understand the pathophysiological mechanism of the postmenopausal osteoporosis, a series of studies were conducted at the whole animal and cellular level. At the whole animal level, female Sprague-Dawley rats were ovariectomized (Ovx) or sham-operated (control) at 18 weeks and the entire skeleton obtained at 24 weeks (baseline) or after an additional 31 day (28 week) interval on a normal (1.0%) or deficient (0.02%) calcium diet. Ovx rats on the normal calcium diet showed a 42% increase in whole body bone resorption and a 70% …

Response Properties Of Tibial Campaniform Sensilla On The Cockroach Leg In Restrained Preparations And Freely Moving Animals, Angela L. Ridgel

Theses, Dissertations and Capstones

The ability to detect changes in forces is important for effective use of a leg in posture and locomotion. This thesis examines how forces are detected in the legs of cockroaches by tibial campaniform sensilla. Campaniform sensilla are mechanoreceptors that encode forces through ovoid cuticular caps embedded in the exoskeleton. The tibial sensilla are unique in that they consist of two subgroups with mutually perpendicular cap orientations.

We characterized the responses of the tibial receptors in restrained preparations by applying forces to the leg at controlled magnitudes and rates. The tibial sensilla, as a group, were sensitive to increasing and …

Mitochondrial 16s Rrna Gene As A Tool For Scorpion Systematics, Mark D. Barker

Theses, Dissertations and Capstones

The use of morphological analysis in the classification and phylogeny of scorpions often has led to confusion and controversy. In order to clarify these problems other character sets should be considered for analysis. One such tool is the use of molecular comparisons between organisms. This study used a section of the mitochondrial 16S ribosomal RNA gene. This technique was applied to investigate four scorpion genera belonging to three families: Hadruru, Anuroctonus (luridae), Euscorpius (Euscorpiidae), and Centruroides (Buthidae). Total DNA was extracted with a standard phenol/chloroform method; a gene of interest was amplified using PCR; and resulting DNA sequence analysis was …

Interaction Of Laminin Beta2 With Eps8 And Desmin In Muscle Cells, Luke Xueliang Cui

Theses, Dissertations and Capstones

Laminin β2 is localized to the synaptic basal lamina (BL) in muscle and glomerular BL in kidney. To find interacting proteins, a mouse kidney cDNA library was screened with domain I of rat laminin β2 using the yeast two-hybrid system. Fifteen positive clones were found. The DNA sequence of three each of these clones corresponded to the phosphoprotein Eps8 (epidermal growth factor receptor pathway substrate clone 8, which is tyrosine phosphorylated after EGF stimulation) and the intermediate filament (IF) protein keratin. To determine if this interaction occurred in muscle cells, C2 cells transfected with the rat …

The Role Of Retinoids And Protein Kinase C In Vascular Smooth Muscle Cell Proliferation And Function, Shaoshan Wang

Theses, Dissertations and Capstones

The structural and functional changes in the arterial wall of humans and animals accompany the development of hypertension and atherosclerosis. Hence, great interest exists in the mechanisms that govern vascular smooth muscle cell contraction and proliferation. Rat aortic rings maintained in organ culture show a significant loss of contractility; whereas, vascular smooth muscle cells in culture quickly dedifferentiate into a secretory phenotype with a marked capacity for proliferation. The addition of all-trans retinoic acid (RA) and retinol to the plasma-containing medium preserved the contractile response of cultured vessels. Removal of endothelium prior to culture abolished these effects. All-trans retinoic acid …

Clarification Of The Hybrid Origin Of Carex X Deamii Herm. (Cyperaceae) Based On Macro And Micro Morphological Characters, Paul B. Marcum

Theses, Dissertations and Capstones

The origin of Carex x deamii Herm. has been in question ever since it was named to science in 1938. Collections of the hybrid have always been associated with C. shortiana Dewey and either C. typhina Michx. or C. squarrosa L.. C. typhina and C. squarrosa are closely related taxa (Section Squarrosae) and often are morphologically similar. Because of this similarity, determining the correct parental species to the hybrid has been extremely difficult. It is known that the hybrid is sterile and only reproduces asexually. Pollen was analyzed to ascertain the viability of all four taxa. This study utilizes both …