Cell and Developmental Biology Commons^™

Early Skeletal Morphogenesis: The Role Of Pharyngeal Endoderm Derived Signaling During Specification Of Craniofacial Skeleton And Interplay Of Mechanical Forces And Molecular Signaling During Skeletal Condensation, Poulomi Ray

All Dissertations

This study examines the fundamental principles governing the early
stages of skeletal morphogenesis. I specifically investigate the specification of mesenchymal progenitor cells to a skeletal fate. Using explant culture system, I determine that the pharyngeal endoderm was sufficient, but not necessary for specifying pre-chondrogenic identity. FGF signaling is both sufficient and required for specification of Sox9 expression and specification of prechondrogenic identity, as demonstrated by the addition of recombinant FGF protein or the FGF receptor inhibitor (SU5402) to explanted tissue, respectively. However, FGF signaling cannot maintain Sox9 expression or initiate the chondrogenic program as indicated by the absence of Col2a1 …

Nuclear Localization Of Cpi-17, A Protein Phosphatase-1 Inhibitor Protein, Affects Histone H3 Phosphorylation And Corresponds To Proliferation Of Cancer And Smooth Muscle Cells., Masumi Eto, Jason A Kirkbride, Rishika Chugh, Nana Kofi Karikari, Jee In Kim

Department of Molecular Physiology and Biophysics Faculty Papers

CPI-17 (C-kinase-activated protein phosphatase-1 (PP1) inhibitor, 17kDa) is a cytoplasmic protein predominantly expressed in mature smooth muscle (SM) that regulates the myosin-associated PP1 holoenzyme (MLCP). Here, we show CPI-17 expression in proliferating cells, such as pancreatic cancer and hyperplastic SM cells. Immunofluorescence showed that CPI-17 was concentrated in nuclei of human pancreatic cancer (Panc1) cells. Nuclear accumulation of CPI-17 was also detected in the proliferating vascular SM cell culture and cells at neointima of rat vascular injury model. The N-terminal 21-residue tail domain of CPI-17 was necessary for the nuclear localization. Phospho-mimetic Asp-substitution of CPI-17 at Ser12 attenuated the nuclear …

Intrinsic Apoptotic Pathway: Effects Of Calcium On Murine Cytochrome C Release In Brain And Liver Mitochondria, Dane M. Edwards

Senior Honors Theses

A cell may use one of three main apoptotic pathways leading to programmed cell death: the extrinsic pathway, the perforin/granzyme pathway and the intrinsic pathway. The most pertinent to this discussion is the intrinsic pathway, which utilizes the mitochondria as an essential intermediary. Mitochondria’s primary function in relation to this pathway is the subsequent release of pro-apoptotic factors including cytochrome c, which activate a caspase cascade leading to the death of the cell. Cytochrome c is released partly due to an increase in cytosolic calcium levels. Two methods of the release of cytochrome c have been proposed. The first is …

In Vitro Anticancer Screening Of 24 Locally Used Nigerian Medicinal Plants, Saudat Adamson Fadeyi, Olugbeminiyi O. Fadeyi, Adedeji A. Adejumo, Cosmas Okoro, Elbert L. Myles

Chemistry Faculty Research

Background

Plants that are used as traditional medicine represent a relevant pool for selecting plant candidates that may have anticancer properties. In this study, the ethnomedicinal approach was used to select several medicinal plants native to Nigeria, on the basis of their local or traditional uses. The collected plants were then evaluated for cytoxicity.

Methods

The antitumor activity of methanolic extracts obtained from 24 of the selected plants, were evaluated in vitro on five human cancer cell lines.

Results

Results obtained from the plants screened indicate that 18 plant extracts of folk medicine exhibited promising cytotoxic activity against human carcinoma …

Hsv In Cervical Cancer, Bruce Bejcek

Faculty Research and Creative Activities Award (FRACAA)

No abstract provided.

Cellular Life Beyond An Individual’S Death, Emily Schmitt

CAHSS Intellectual Conversations

One of the most famous examples of cell lines surviving long after a person has died comes from a tumor found in the body of Henrietta Lacks, who died of cervical cancer in 1950. The study of approximately 50 million tons of her cells, which would conservatively equal the amount of cells in one billion people, has resulted in nearly 11,000 patents, including the polio vaccine. These cells have become a focus of public attention since Rebecca Skloot’s 2010 book, The Immortal Life of Henrietta Lacks. This talk will discuss what happens to cells once they leave our bodies, …

Defining The Efficacy Of Aortic Root Enlargement Procedures: A Comparative Analysis Of Surgical Techniques, Katie L. Losenno, Jill J. Gelinas, Marjorie Johnson, Michael W. A. Chu

Anatomy and Cell Biology Publications

Background: Aortic root enlargement (ARE) procedures are believed to allow implantation of larger valve prostheses; however, little evidence exists to support the specific efficacy of various techniques. Methods: Using a cadaveric model, 20 adult (72.4 +/- 15.3 years) hearts were stratified into 4 groups based on annular diameter: <20 mm, 20-22 mm, 22-24 mm, and >24 mm. Each heart underwent an aortic valve replacement following a Nicks, Manougian, aortoventriculoplasty and modified Bentall procedure, with appropriate reversals between procedures. Results: All 4 groups experienced similar increases in annular diameter (P = 0.43) and prosthesis size implanted (P = 0.51) with each enlargement technique. The Nicks, Manougian, modified Bentall …

Improved Tolerance To Freeze And Salt Stress In Mitochondria Of Drosophilia Melanogaster Cells Conferred By Lea Protein, Matthew R. Marunde

Undergraduate Honors Theses

Mechanisms that govern anhydrobiosis involve the accumulation of highly hydrophilic macromolecules, such as late embryogenesis abundant (LEA) proteins. Group 1 LEA proteins comprised of 181 (AfLEA1.1) and 197 (AfLEA1.3) amino acids were cloned from embryos of Artemia franciscana and expressed in Drosophila melanogaster cells (Kc167). Confocal microscopy revealed accumulations of green fluorescence protein (GFP) and AfLEA1.3 constructs in the mitochondria (AfLEA1.3-GFP) and AfLEA1.1-GFP constructs in the cytoplasm. In the presence of mixed substrates, oxygen consumption was similar for permeabilized Kc167 control and Kc167-AfLEA1.3 cells. Acute titrations of NaCl (up to 500 mM) led to successive drops in oxygen flux of …

Evidence For The Requirement Of 14-3-3eta (Ywhah) In Meiotic Spindle Assembly During Mouse Oocyte Maturation, Santanu De, Douglas Kline

Biology Faculty Articles

Background

The 14-3-3 (YWHA) proteins are central mediators in various cellular signaling pathways regulating development and growth, including cell cycle regulation. We previously reported that all seven mammalian 14-3-3 isoforms are expressed in mouse oocytes and eggs and that, 14-3-3η (YWHAH) accumulates and co-localizes in the region of meiotic spindle in mouse eggs matured in vivo. Therefore, we investigated the role of 14-3-3η in spindle formation during mouse oocyte maturation.

Results

Examination of oocytes matured in vitro demonstrated that 14-3-3η accumulates in both meiosis I and II spindles. To explore if 14-3-3η interacts directly with α-tubulin in meiotic spindles, …

Effects Of Synthetic Histone Deacetylase Inhibitors On 3t3l1 Preadipocyte Adipogenic Differentiation Potential, Vikramjeet Singh

Honors Capstones

No abstract provided.

A Myosin-Va Tail Fragment Sequesters Dynein Light Chains Leading To Apoptosis In Melanoma Cells, Tc Izidoro-Toledo, Ac Borges, Dd Araujo, Dps Leitao Mazzi, Fo Nascimento Junior, Jf Sousa, Cp Alves, Apb Paiva, Dm Trindade, Ev Patussi, Pablo M. Peixoto, Kw Kinnally, Em Espreafico

Publications and Research

Previous studies proposed that myosin-Va regulates apoptosis by sequestering pro-apoptotic Bmf to the actin cytoskeleton through dynein light chain-2 (DLC2). Adhesion loss or other cytoskeletal perturbations would unleash Bmf, allowing it to bind and inhibit pro-survival Bcl2 proteins. Here, we demonstrated that over expression of a myosin-Va medial tail fragment (MVaf) harboring the binding site for DLC2 dramatically decreased melanoma cell viability. Morphological and molecular changes, including surface blebbing, mitochondrial outer membrane permeabilization, cytochrome-c and Smac release, as well as caspase-9/-3 activation and DNA fragmentation indicated that melanoma cells died of apoptosis. Immobilized MVaf interacted directly with DLCs, but complexed …

Expression Of Growth Arrest And Dna Damage Protein 45-Alpha (Gadd45-Alpha) And The Ccaat/Enhancer Binding Protein-Delta (C/Ebp-Delta) In Fishes Exposed To Heat And Hypoxia, Daniel Omar Hassumani

Dissertations and Theses

The cellular stress response (CSR) is one of the most highly conserved mechanisms among all organisms. Cellular stress can be defined as damage or the threat of damage to proteins, macromolecules and/or DNA. The response to damage can involve cell cycle regulation, protein chaperoning, DNA repair or, if macromolecular damage is too severe, apoptotic mechanisms can be initiated. This thesis details experiments that were designed to examine the cellular response to non-lethal environmental stressors at the protein level, using two fish species as study models. Two proteins that can cause cell cycle arrest and apoptosis mechanisms were examined. Expression of …

Metabolic Rescue Of “Glucose Addicted” Cancer Cells In Vitro, Paolo Vignali

Pursuit - The Journal of Undergraduate Research at The University of Tennessee

Transformations in the glycolytic metabolism of neoplasms modulate their robust cellular division. This characteristic leads to an “addiction” to glucose for continued proliferation and viability. This study investigated whether glucose metabolites could rescue cellular viability in glucose-starvation conditions, a model of the inter-tumoral nutrient-deficient environment. Findings illustrated potential cellular viability rescue with pyruvate addition in glucose-deprived conditions, yet the same potential was not observed with lactic acid, a metabolite that exists at characteristically high concentrations within the intertumoral microenvironment. These results could implicate a predominance of certain metabolic pathways in nutrient-starved cells. Molecular transport capacities across plasma membranes are tied …

Molecular Imaging To Target Transplanted Muscle Progenitor Cells, Kelly Gutpell, Rebecca Mcgirr, Lisa Hoffman

Anatomy and Cell Biology Publications

Duchenne muscular dystrophy (DMD) is a severe genetic neuromuscular disorder that affects 1 in 3,500 boys, and is characterized by progressive muscle degeneration(1, 2). In patients, the ability of resident muscle satellite cells (SCs) to regenerate damaged myofibers becomes increasingly inefficient(4). Therefore, transplantation of muscle progenitor cells (MPCs)/myoblasts from healthy subjects is a promising therapeutic approach to DMD. A major limitation to the use of stem cell therapy, however, is a lack of reliable imaging technologies for long-term monitoring of implanted cells, and for evaluating its effectiveness. Here, we describe a non-invasive, real-time approach to evaluate the success of myoblast …

Large Area Periodic, Systematically Changing, Multishape Nanostructures By Laser Interference Lithography And Cell Response To These Topographies, Erden Ertorer, Fartash Vasefi, Joel Keshwah, Mohamadreza Najiminaini, Christopher Halfpap, Uwe Langbein, Jeffrey J. L. Carson, Douglas W. Hamilton, Silvia Mittler

Anatomy and Cell Biology Publications

The fabrication details to form large area systematically changing multishape nanoscale structures on a chip by laser interference lithography (LIL) are described. The feasibility of fabricating different geometries including dots, ellipses, holes, and elliptical holes in both x- and y- directions on a single substrate is shown by implementing a Lloyd's interferometer. The fabricated structures at different substrate positions with respect to exposure time, exposure angle and associated light intensity profile are analyzed. Experimental details related to the fabrication of symmetric and biaxial periodic nanostructures on photoresist, silicon surfaces, and ion milled glass substrates are presented. Primary rat calvarial osteoblasts …

Matrix Adhesion Polarizes Heart Progenitor Induction In The Invertebrate Chordate Ciona Intestinalis, J. Norton, J. Cooley, A. Islam, C. D. Cota, Bradley Justin Davidson , '91

Biology Faculty Works

Cell-matrix adhesion strongly influences developmental signaling. Resulting impacts on cell migration and tissue morphogenesis are well characterized. However, the in vivo impact of adhesion on fate induction remains ambiguous. Here, we employ the invertebrate chordate Ciona intestinalis to delineate an essential in vivo role for matrix adhesion in heart progenitor induction. In Ciona pre-cardiac founder cells, invasion of the underlying epidermis promotes localized induction of the heart progenitor lineage. We found that these epidermal invasions are associated with matrix adhesion along the pre-cardiac cell/epidermal boundary. Through targeted manipulations of RAP GTPase activity, we were able to manipulate pre-cardiac cell-matrix adhesion. …

Molecular Immunologic Correlates Of Spontaneous Latency In A Rabbit Model Of Pulmonary Tuberculosis, Selvakumar Subbian, Paul O'Brien, Nicole L. Kushner, Guibin Yang, Liana Tsenova, Blas Peixoto, Nirmalya Bandyopadhyay, Joel S. Bader, Petros C. Karakousis, Dorothy Fallows, Gilla Kaplan

Publications and Research

Background: Infection of humans with Mycobacterium tuberculosis (Mtb) results in latent tuberculosis infection (LTBI) in 90-95% of immune competent individuals, with no symptoms of active disease. The World Health Organization estimates that 1.5 billion people have LTBI, which can reactivate in the setting of waning host immunity, posing a threat to global TB control. Various animal models have been used to study the pathogenesis of TB. However, besides nonhuman primates, rabbits are the only animal model that fully recapitulates the pathological features of human TB, including progressive disease with necrosis and cavitation or establishment of spontaneous latency.

Results: We …

Microbial Biomineralization Of Iron, Wen Fang

Dissertations and Theses

Iron is a common cation in biomineral sand; it is present for example in magnetite produced by magnetotactic bacteria and in iron sulfides produced by sulfate reducing microorganisms. The work presented in this thesis focused on two types of microorganisms capable of forming iron biominerals. In the first project I have studied the effect of O2 on the respiratory physiology and the formation of magnetosomes by Magnetospirillum magneticum AMB-1. In the second project I have studied the relationship between olivine and the activity of dissimilatory sulfate reducing (DSR) microorganisms. For the first project, I grew cells of AMB-1 in cultures …

Glucosensing, Hypoglycemia, And The Brain, Candace Reno

All Theses and Dissertations (ETDs)

Glucose homeostasis can be peripherally and centrally mediated. Within the periphery, GLUT4 in muscle and adipose tissue is required for normal glucose homeostasis and disruption of GLUT4 translocation in these tissues leads to glucose intolerance and insulin resistance, hallmarks of Type 2 diabetes. Within the brain, specialized glucose sensing neurons regulate whole body glucose homeostasis. Unlike GLUT4's role in the periphery, brain GLUT4 regulation of glucose tolerance is not well defined. Understanding how brain glucose sensing, mediated by GLUT4, regulates glucose tolerance and insulin sensitivity will aid in the development of treatment strategies to protect against the pathogenesis of diabetes. …

The Herpesvirus Vp1/2 Protein Is An Effector Of Dynein-Mediated Capsid Transport And Neuroinvasion, Sofia V. Zaichick, Kevin P. Bohannon, Ami Hughes, Patricia J. Sollars, Gary E. Pickard, Gregory A. Smith

School of Veterinary and Biomedical Sciences: Faculty Publications

Microtubule transport of herpesvirus capsids from the cell periphery to the nucleus is imperative for viral replication and, in the case of many alphaherpesviruses, transmission into the nervous system. Using the neuroinvasive herpesvirus, pseudorabies virus (PRV), we show that the viral protein 1/2 (VP1/2) tegument protein associates with the dynein/dynactin microtubule motor complex and promotes retrograde microtubule transport of PRV capsids. Functional activation of VP1/2 requires binding to the capsid protein pUL25 or removal of the capsid-binding domain. A proline-rich sequence within VP1/2 is required for the efficient interaction with the dynein/ dynactin microtubule motor complex as well as for …