Laboratory and Basic Science Research Commons^™

Observing The Unwatchable Through Acceleration Logging Of Animal Behavior, Danielle D. Brown, Roland Kays, Martin Wikelski, Rory Wilson, A. Peter Klimley

Methodology and Animal Models in Research

Behavior is an important mechanism of evolution and it is paid for through energy expenditure. Nevertheless, field biologists can rarely observe animals for more than a fraction of their daily activities and attempts to quantify behavior for modeling ecological processes often exclude cryptic yet important behavioral events. Over the past few years, an explosion of research on remote monitoring of animal behavior using acceleration sensors has smashed the decades-old limits of observational studies. Animal-attached accelerometers measure the change in velocity of the body over time and can quantify fine-scale movements and body postures unlimited by visibility, observer bias, or the …

Investigation Of The Role Of Tor And Nutrient Restriction On Lifespan Of Arabidopsis Thaliana, Debashree Banerjee

Dissertations, Masters Theses, Capstones, and Culminating Projects

The Target of Rapamycin (TOR) kinase is evolutionarily conserved and is a key regulator, controlling many cellular functions to promote survival and growth in all eukaryotes. Several model organisms have indicated a relationship between TOR signaling and life expectancy, such as yeast, fruit flies and mice. Abundant nutrient availability promotes rapid growth and development, whereas depletion of nutrients reduces the activity of the pathways that are involved in growth and nutrient processing. This reduction in activity increases life expectancy, which is supported by evolutionary theories. The confirmed presence of the TOR homolog (a major regulator of growth and cell proliferation) …

Developmental And Molecular Functions Of Plakophilin-3, William A. Munoz

Dissertations and Theses (Open Access)

Plakophilin-3, the less studied member of the plakophilin-catenin subfamily, and the larger catenin family, binds directly to desmosomal cadherin cytoplasmic domains and enhances desmosome formation and stability. In mammals, plakophilin-3 is expressed at the highest levels in desmosome-enriched tissues such as epithelia, with the knock-out in mice producing corresponding reductions in ectodermal integrity. In tissue, cellular and intracellular contexts where plakophilin-3 is not at the desmosomal plaque, little is known about its functions in the cytoplasm or nucleus, where it also localizes.

My work employed embryos of the amphibian, Xenopus laevis, to examine plakophilin-3’s developmental roles. I first evaluated …

The Roles Of Phenotypic Plasticity And Genotypic Specialization In High Altitude Adaptation, Danielle M. Tufts

School of Biological Sciences: Dissertations, Theses, and Student Research

In vertebrates living at high altitude, arterial hypoxemia may be ameliorated by reversible changes in the oxygen-carrying capacity of the blood (regulated by erythropoiesis) and/or changes in blood–oxygen affinity (regulated by allosteric effectors of hemoglobin function). These hematological traits often differ between taxa that are native to different elevational zones, but it is often unknown whether the observed physiological differences reflect fixed, genetically based differences or environmentally induced acclimatization responses (phenotypic plasticity). Here, we report measurements of hematological traits related to blood–O₂ transport in populations of deer mice (Peromyscus maniculatus) that are native to high- and low-altitude …

Cytotoxicity In The Age Of Nano: The Role Of Fourth Period Transition Metal Oxide Nanoparticle Physicochemical Properties, Charles C. Chusuei, Chi-Heng Wu, Shravan Mallavarapu, Fang Yao Stephen Hou, Chen-Ming Hsu, Jeffrey G. Winiarz, Robert S. Aronstam, Yue-Wern Huang

Clinical Lab Sciences Faculty Research and Publications

A clear understanding of physicochemical factors governing nanoparticle toxicity is still in its infancy. We used a systematic approach to delineate physicochemical properties of nanoparticles that govern cytotoxicity. The cytotoxicity of fourth period metal oxide nanoparticles (NPs): TiO₂, Cr₂O₃, Mn₂O₃, Fe₂O₃, NiO, CuO, and ZnO increases with the atomic number of the transition metal oxide. This trend was not cell-type specific, as observed in non-transformed human lung cells (BEAS-2B) and human bronchoalveolar carcinoma-derived cells (A549). Addition of NPs to the cell culture medium did not significantly …

Incorporating New Technologies Into Toxicity Testing And Risk Assessment: Moving From 21st Century Vision To A Data-Driven Framework, Russell S. Thomas, Martin A. Philbert, Scott Auerbach, Barbara A. Wetmore, Michael J. Devito, Ila Cote, J. Craig Rowlands, Maurice Whelan, Sean M. Hays, Melvin E. Andersen, M.E. (Bette) Meek, Lawrence W. Reiter, Jason C. Lambert, Harvey J. Clewell Iii, Martin L. Stephens, Q. Jay Zhao, Scott C. Wesselkamper, Lynn Flowers, Edward W. Carney, Tim P. Pastoor, Dan D. Petersen, Carole L. Yauk, Andy Nong

in Vitro Research Models Collection

Based on existing data and previous work, a series of studies is proposed as a basis toward a pragmatic early step in transforming toxicity testing. These studies were assembled into a data-driven framework that invokes successive tiers of testing with margin of exposure (MOE) as the primary metric. The first tier of the framework integrates data from high-throughput in vitro assays, in vitro-to-in vivo extrapolation (IVIVE) pharmacokinetic modeling, and exposure modeling. The in vitro assays are used to separate chemicals based on their relative selectivity in interacting with biological targets and identify the concentration at which these interactions …

Facile Synthesis, Characterization, And Antimicrobial Activity Of Cellulose-Chitosan-Hydroxyapatite Composite Material: A Potential Material For Bone Tissue Engineering, Tamutsiwa M. Mututuvari, April L. Harkins, Chieu D. Tran

Clinical Lab Sciences Faculty Research and Publications

Hydroxyapatite (HAp) is often used as a bone-implant material because it is biocompatible and osteoconductive. However, HAp possesses poor rheological properties and it is inactive against disease-causing microbes. To improve these properties, we developed a green method to synthesize multifunctional composites containing: (1) cellulose (CEL) to impart mechanical strength; (2) chitosan (CS) to induce antibacterial activity thereby maintaining a microbe-free wound site; and (3) HAp. In this method, CS and CEL were co-dissolved in an ionic liquid (IL) and then regenerated from water. HAp was subsequently formed in situ by alternately soaking [CEL+CS] composites in aqueous solutions of CaCl_{2 …}

Raman Spectroscopy As A Teaching Tool In The Forensic Science And Chemistry Courses, Zhaohua Dai, Elmer-Rico E. Mojico

Cornerstone 3 Reports : Interdisciplinary Informatics

No abstract provided.

Bacterial Transcription Factors Grea, Greb And Dksa: Characterization Of Their Interaction With Rna Polymerase And Molecular Mechanism Of Action, Andrey Parshin

Graduate School of Biomedical Sciences Theses and Dissertations

The activity of RNA polymerase (RNAP), the key enzyme of transcription process, is regulated by a large number of transcription factors acting at each step of the transcription cycle. Our research project is focused on studying the structure and function of bacterial transcript cleavage factors GreA, GreB and the stringent response regulator DksA. Gre factors bind in the secondary channel of RNAP in backtracked (inactive) ternary elongation complexes and stimulate the enzyme's intrinsic RNase activity, which is required for suppression of transcription pause and arrest, enhancement of transcription fidelity, and efficient promoter escape. DksA also acts through the RNAP secondary …

Life Support: Long Term Storage Of Solid Waste In An Enclosed Membrane System, Emmy O. Trieu, Michael Flynn, Rocco Mancinelli

STAR Program Research Presentations

In deep space missions, maintaining life support is of the utmost priority. In such a closed system, human waste must be stored and treated. Simulated feces (ersatz) will be inoculated with microbes representing normal fecal flora, mixed with urine brine and shredded refuse of products typically used in space missions. Composting methods often use alternating layers of waste with scraps of carboniferous materials (finely shredded refuse). By preparing membrane bags with a homogenized ersatz and carboniferous refuse mixture and membrane bags with alternating layers of ersatz and carboniferous refuse, it may be possible to monitor anaerobic thermophillic digestion of the …

Modulation Of Bax/Bak Dependent Apoptosis By Sirtuin 3 And Mitochondrial Permeability Transition By Sirtuin 4, Manish Verma

Graduate School of Biomedical Sciences Theses and Dissertations

Mitochondria are dynamic organelles that regulate a myriad of cellular functions, including energy production and metabolic regulation. Mitochondria are also a critical regulator of cell death signaling cascades modulating both apoptotic and necrotic cell death. However, what determines which cell death pathway is activated is still unclear. The mitochondrial/intrinsic pathway of apoptosis is dependent on the activation of pro-apoptotic proteins, Bax and Bak, which induce mitochondrial outer membrane permeabilization (MOMP). Once the integrity of outer mitochondrial membrane (OMM) is compromised, pro-apoptotic intermembrane space proteins like cytochrome c, Smac/Diablo, Omi/HtrA2 and AIF are released into the cytoplasm, which activates the post-mitochondrial …

Female Rats Display Enhanced Rewarding Effects Of Ethanol That Are Hormone Dependent, Laura O'Dell

Selected Works Temporary Series

Background: Ethanol (EtOH) abuse is a major health and economic concern, particularly for females who appear to be more sensitive to the rewarding effects of EtOH. This study compared sex differences to the rewarding and aversive effects of EtOH using place-conditioning procedures in rats. Methods: Separate groups of adult (male, female, ovariectomized [OVX] female) and adolescent (male and female) rats received EtOH (0, 0.5, 1.0, 2.0 or 2.5 g/kg; i.p.) and were confined to their initially nonpreferred side of our conditioning apparatus for 30 minutes. On alternate days, they received saline and were confined to the other side. Following 5 …

Med13p Prevents Stress-Independent Mitochondrial Hyperfragmentation And Aberrant Apoptosis Activation In Saccharomyces Cerevisiae By Controlling Cyclin C Nuclear Localization, Svetlana Khakhina

Graduate School of Biomedical Sciences Theses and Dissertations

During aging, and as a result of environmental changes, cells are exposed to elevated levels of reactive oxygen species (ROS). High ROS levels induce lipid oxidation, protein aggregation, mitochondrial hyperfragmentation, DNA damage and programmed cell death (PCD), also called apoptosis. PCD is a highly regulated process and its misregulation has been linked to neurodegenerative diseases and cancer development.

Our hypothesis is that cyclin C plays a role in the initiation of apoptosis. During normal conditions, cyclin C represses the transcription of stress response genes (SRG). In response to stress, cyclin C translocates to the cytoplasm where it facilitates mitochondrial hyperfragmentation …

Female Rats Display Enhanced Rewarding Effects Of Ethanol That Are Hormone Dependent, Laura O'Dell

Departmental Papers (Psych)

Background: Ethanol (EtOH) abuse is a major health and economic concern, particularly for females who appear to be more sensitive to the rewarding effects of EtOH. This study compared sex differences to the rewarding and aversive effects of EtOH using place-conditioning procedures in rats. Methods: Separate groups of adult (male, female, ovariectomized [OVX] female) and adolescent (male and female) rats received EtOH (0, 0.5, 1.0, 2.0 or 2.5 g/kg; i.p.) and were confined to their initially nonpreferred side of our conditioning apparatus for 30 minutes. On alternate days, they received saline and were confined to the other side. Following 5 …

Characterization Of Jak, Stat, And Src Interactions In Head And Neck Squamous Cell Carcinoma, Reshma Jaseja, Reshma Jaseja

Dissertations and Theses (Open Access)

Recurrence of Head and Neck Squamous Cell Carcinoma (HNSCC) is common; thus, it is essential to improve the effectiveness and reduce toxicity of current treatments. Proteins in the Src/Jak/STAT pathway represent potential therapeutic targets, as this pathway is hyperactive in HNSCC and it has roles in cell migration, metastasis, proliferation, survival, and angiogenesis. During short-term Src inhibition, Janus kinase (Jak) 2, and signal transducer and activator of transcription (STAT) 3 and STAT5 are dephosphorylated and inactivated. Following sustained Src inhibition, STAT5 remains inactive, but Jak2 and STAT3 are reactivated following their early inhibition. To further characterize the mechanism of this …

Coccidia Of Gerbils From Mongolia, Ethan Thomas Jensen

School of Biological Sciences: Dissertations, Theses, and Student Research

In this study, gerbils collected in the Mongolia over the summers of 2009-2012 were examined for coccidia. In total, 171 gerbils of three species from 22 localities were examined for coccidia. Coccidian oocysts were identified from 21 gerbils, but those found in 1 of those gerbils were probably pseudoparasites of the host from which they were recovered. From the remaining 20 gerbils, 7 morphotypes of Eimeria and 1 morphotype of Isospora were identified. Four of the 7 morphotypes of Eimeria were attributed to new species which were described in this study. In addition, 10 previously described species of Eimeria were …

Novel Roles Of Replication Protein A Phosphorylation In Cellular Response To Dna Damage, Moises A. Serrano

Electronic Theses and Dissertations

Human replication protein A (RPA) is an eukaryotic single-stranded DNA binding protein directly involved in a variety of DNA metabolic pathways including replication, recombination, DNA damage checkpoints and signaling, as well as all DNA repair pathways. This project presents 2 novel roles of RPA in the cellular response to DNA damage. The first elucidates the regulation of RPA and p53 interaction by DNA-dependent protein kinase (DNA-PK), ataxia telangiectasia mutated (ATM) and ATM- and Rad3-related (ATR) in homologous recombination (HR). HR and nonhomologous end joining (NHEJ) are 2 distinct DNA double-stranded break (DSB) repair pathways. Here, we report that DNA-PK, the …

Diabetes Mellitus And Hypercholesterolemia Are Risk Factors For Alzheimer’S Disease And Appear To Affect The Integrity Of The Blood Brain Barrier, Jacqueline Dash

Graduate School of Biomedical Sciences Theses and Dissertations

Studies have shown that the vascular risk factors common to diabetes mellitus and hypercholesterolemia are also risk factors for Alzheimer’s disease (AD). It is currently unknown how these diseases are associated with AD, but they may cause a leak in the blood brain barrier (BBB), which is one of the hallmarks of AD. In this preliminary study, over 150 pig brain slides were tested for the expression levels of tight junction proteins occludin and claudin V in the BBB microvasculature. There were three groups of pig brains used in this study namely, control pigs, pigs with diabetes mellitus and hypercholesterolemia …

A Mechanistic Hypothesis Of The Factors That Enhance Vulnerability To Nicotine Use In Females, Laura O'Dell

Selected Works Temporary Series

Women are particularly more vulnerable to tobacco use than men. This review proposes a unifying hypothesis that females experience greater rewarding effects of nicotine and more intense stress produced by withdrawal than males. We also provide a neural framework whereby estrogen promotes greater rewarding effects of nicotine in females via enhanced dopamine release in the nucleus accumbens (NAcc). During withdrawal, we suggest that corticotropin-releasing factor (CRF) stress systems are sensitized and promote a greater suppression of dopamine release in the NAcc of females versus males. Taken together, females display enhanced nicotine reward via estrogen and amplified effects of withdrawal via …

Can Acute Dermal Systemic Toxicity Tests Be Replaced With Oral Tests? A Comparison Of Route-Specific Systemic Toxicity And Hazard Classifications Under The Globally Harmonized System Of Classification And Labelling Of Chemicals (Ghs), Nigel P. Moore, David J. Andrew, Donald L. Bjerke, Stuart Creton, David Dreher, Thomas Holmes, Pilar Prieto, Troy Seidle, Tim G. Rowan

Experimentation Collection

Acute systemic toxicity data (LD50 values) and hazard classifications derived in the rat following oral administration and dermal application have been analysed to examine whether or not orally-derived hazard classification or LD50 values can be used to determine dermal hazard classification. Comparing the oral and dermal classifications for 335 substances derived from oral and dermal LD50 values respectively revealed 17% concordance, and indicated that 7% of substances would be classified less severely while 76% would be classified more severely if oral classifications were applied directly to the dermal route. In contrast, applying the oral LD50 values within the dermal classification …

A Mechanistic Hypothesis Of The Factors That Enhance Vulnerability To Nicotine Use In Females, Laura O'Dell

Departmental Papers (Psych)

Women are particularly more vulnerable to tobacco use than men. This review proposes a unifying hypothesis that females experience greater rewarding effects of nicotine and more intense stress produced by withdrawal than males. We also provide a neural framework whereby estrogen promotes greater rewarding effects of nicotine in females via enhanced dopamine release in the nucleus accumbens (NAcc). During withdrawal, we suggest that corticotropin-releasing factor (CRF) stress systems are sensitized and promote a greater suppression of dopamine release in the NAcc of females versus males. Taken together, females display enhanced nicotine reward via estrogen and amplified effects of withdrawal via …

Purification And Characterization Of Oxidation-Resistant Ribonuclease Inhibitor Variants, Alec W. Uebersohn

Lawrence University Honors Projects

Ribonuclease inhibitor (RI) is an intracellular mammalian protein which binds vertebrate-specific ribonucleases; this interaction is one of the tightest non-covalent interactions yet discovered. The biological activity of RI is poorly understood, but it is thought to regulate the biological functions of ribonucleases, which include initiating blood vessel growth, maintaining neuron viability, attacking pathogens, and mediating cell stress responses. RI is also involved in pathways unrelated to ribonucleases, including interactions with Drosha and PTEN, an anti-tumor protein.

One of the defining characteristics of RI is its oxidation sensitivity, a result of its unusually high cysteine content. The oxidation of RI is …

Validation Of Antibodies Used To Study Hypoxia Inducible Factors In Two Species Of Fundulus, Jenna D. Hill

LSU New Orleans Theses and Dissertations

Hypoxia inducible factors (HIFs) are transcription factors and the master regulators of oxygen-dependent gene expression in animals. The focus of this thesis is the distribution of HIF protein in tissues of the fish Fundulus heteroclitus and F. grandis, two widespread species that occur in naturally hypoxic waters. Polyclonal antibodies against HIF-1α, HIF-2α, and HIF-3α were tested on proteins made in vitro and on extracts made from several tissues of normoxic and hypoxic fish. Antibodies against HIF-1α and 3α bound specifically to full length protein made in vitro, and produced bands on western blots of nuclear extracts of near …

Metronidazole Effects On Hindgut Microflora Residing Within The Subterranean Termite Reticulitermes Flavipes, Alexandra Gjevre

Morehead State Theses and Dissertations

A thesis presented to the faculty of the College of Science and Technology at Morehead State University in partial fulfillment of the requirements for the Degree of Master of Science by Alexandra Gjevre on May 1, 2013.

The Effects Of Chronic Cadmium Exposure On Breast Cancer Progression, Esmeralda Ponce

Dissertations, Masters Theses, Capstones, and Culminating Projects

Cadmium is an environmental contaminant that enters the body usually through diet or cigarette smoke. Exposure to cadmium has been associated with the development of breast cancer. Recent studies have suggested that cadmium functions as an endocrine disruptor and mimics the actions of estrogen in breast cancer cells by activating the receptor (ER) to promote breast cancer cell growth. Although acute cadmium exposure is known to promote estrogen receptor-mediated gene expression associated with growth, the consequence of chronic cadmium exposure has been unclear. Since heavy metals are known to bioaccumulate, it is necessary to understand and elucidate the effects of …

Dual Mtor/Hdac Inhibition: Preclinical Development Of A Novel Breast Cancer Therapy, Mariya Alexandra Yevtushenko

Dissertations, Masters Theses, Capstones, and Culminating Projects

HER2 (human epidermal growth factor receptor-2) and/or estrogen receptor (ER) are overexpressed in ~80% of human breast cancers. Although modern therapeutics (e.g. Trastuzumab, Tamoxifen) target the HER2 and ER receptors, clinical resistance often develops due to activation of downstream signaling pathways, including phosphoinositide 3 kinase-AKT/protein kinase Bmammalian target of rapamycin (PI3K/AKT/mTOR), despite effective upstream receptor inhibition. Activation of this pathway increases ribosome biogenesis and translation of oncogenic mRNAs, which are controlled by intracellular histone deacetylase (HDAC) activity as well. In fact, HDAC inhibitors that have been implemented into clinical practice are capable of overcoming resistance to HER2 and ER targeted …

Development Of A Cellular Model For Morquio A Syndrome, Virgen Guadalupe Sierra

Dissertations, Masters Theses, Capstones, and Culminating Projects

Mucopolysaccharidosis IVA (MPS IVA; Morquio A), is a lysosomal storage disorder characterized by the deficiency of N-acetylgalactosamine-6-sulfatase (GALNS), resulting in the accumulation of glycosaminoglycans (GAGs) such as keratan sulfate (KS) (Northover et al., 1996) and chondroitin-6-sulfate. There is no applicable animal model or appropriate cell lines currently available for the study of MPS IVA. Furthermore, patient-specific material is limited and difficult to obtain, rendering the study of MPSIVA pathogenesis challenging. Thus, the generation of an induced pluripotent stem cell (iPSC) line from patient-specific somatic cells shows potential for further examining the pathogenesis of MPS IVA. With this technology, differentiation protocols …

Enzyme Kinetics Of Recombinant Human Arylsulfatase B (Rhasb), Katherine Marie Cunico

Dissertations, Masters Theses, Capstones, and Culminating Projects

This project seeks to explain the enzyme kinetics of recombinant human Arylsulfate sulfatase B (rhASB) by using an in vitro bioassay and direct hydrolysis substrate assay to approach the problem of substrate limitation in establishing a representative kcat for rhASB. The project also explores the makeup of a possible ASB complex within the cell. Previous published studies have sought to establish a turnover rate for rhASB, but have run into the problem of substrate limitation. Using a direct monosaccharide hydrolysis assay in conjunction with the same natural substrate bioassay, this project will attempt to provide insight to the kcat, the …

Mutations In Parkin (Pdr-1) Results In Increased Reproductive Fitness In C. Elegans, Tracy Ann Barhydt

Dissertations, Masters Theses, Capstones, and Culminating Projects

Parkinson’s disease (PD) is largely an idiopathic disease that includes contributions from both genetic and environmental factors, with aging itself being the largest risk factor. These combined susceptibility factors are believed to accumulatively contribute to initiation and progression of Parkinson’s disease. Although most cases of PD are idiopathic, rare genetic forms of the disease do exist. In these cases, an important unanswered question is why mutations that cause the disease have come to be maintained within the human population? One possibility is based on what is known as the Antagonist Pleiotropic Theory (APT) of Aging, a theory that states that …

Characterization Of Tcl1-Tg:P53-/- Mice That Resemble Human Chronic Lymphocytic Leukemia With 17p-Deletion, Jinyun Liu

Dissertations and Theses (Open Access)

Chronic lymphocytic leukemia (CLL) is the most common adult leukemia in the United Statesand Europe. CLL patients with deletion of chromosome 17p, where the tumor suppressor p53 gene is located, often develop a more aggressive disease with poor clinical outcomes. However, the underlying mechanism remains unclear. In order to understand the underneath mechanism in vivo, I have recently generated mice with E_u-TCL1-Tg:p53^-/- genotype and showed that these mice develop aggressive leukemia that resembles human CLL with 17p deletion. The E_u-TCL1-Tg:p53^-/- mice developed CLL disease at 3-4 months, …