Molecular Biology Commons^™

Effect Of A 10 Day Decrease In Physical Activity On Circulating Angiogenic Cells, Gayatri Guhanarayan

Masters Theses 1911 - February 2014

Circulating angiogenic cells (CACs) are early predictors of cardiovascular health and are inversely proportional to related outcomes. Increased number and function of CACs is seen in healthy individuals compared with individuals with cardiovascular disease (CVD). Exercise increases CAC number and function in CVD populations, through a nitric oxide-mediated mechanism. Inactivity is a growing concern in industrialized nations; it is an independent risk factor for CVD and is linked to increased mortality. The purpose of this study was to understand the effect of reduced physical activity (rPA) on two CAC populations (CFU-Hill and CD34⁺) in highly active individuals. We …

A Test Of The Hypothesis That Environmental Chemicals Interfere With Thyroid Hormone Action In Human Placenta, Katherine Geromini

Masters Theses 1911 - February 2014

Thyroid hormone is essential for normal brain development and recognition of this has led to universal screening of newborns for thyroid function to ensure that circulating levels of thyroid hormone are within a range known to be supportive of normal growth and mental development. Environmental chemicals that interfere with thyroid function are known to inhibit normal growth and mental development. Work from our lab and from labs internationally demonstrates in animal systems that some industrial chemicals such as PCBs, PBDEs, and others may interact with the thyroid hormone receptor(s) in ways that are not predicted by changes in serum thyroid …

Proteolytic Regulation Of Ctra, The Master Regulator Of Cell Cycle In Caulobacter Crescentus, Amber M. Cantin

Masters Theses 1911 - February 2014

Cell cycle progression in Caulobacter crescentus depends on the master regulator, CtrA. During the transition from swarmer to stalk cell (G1 to S phase), CtrA is degraded by the AAA+ protease ClpXP and levels rise again in the predivisional stage. The focus of this work is to explore how cyclic, regulated degradation is controlled. CtrA is known to bind to the origin of replication, thereby suppressing replication, so we first asked if DNA binding had an effect on CtrA stability. CtrA is readily degraded by ClpXP on its own, but when bound to DNA containing the proper binding sites, degradation …

Investigation Of Differential Vector Competence Of Bartonella Quintana In Human Head And Body Lice, Domenic J. Previte

Masters Theses 1911 - February 2014

Human head and body lice are obligatory hematophagous ectoparasites that belong to a single species Pediculus humanus. Only body lice, however, are vectors of the infections gram-negative bacteria Bartonella quintana. Due to their near identical genomes, yet differential vector competency, head and body lice provide an ideal model system to study an insects ability to gain or lose vector competency. Using our in vitro louse rearing system, we have infected both head and body lice with a blood containing B. quintana in order to detect differences in B. quintana proliferation between head and body lice as well as …

Connecting Motors And Membranes: A Quantitative Investigation Of Dynein Pathway Components And In Vitro Characterization Of The Num1 Coiled Coil Domain, Bryan J. St. Germain

Masters Theses 1911 - February 2014

In the budding yeast, Saccharomyces Cerevisiae, dynein, a minus-end directed motor, is involved in nuclear migration and proper orientation of the mitotic spindle during mitosis. Our lab has developed a model that involves the loading of cytoplasmic dynein onto the plus-end of astral microtubules through interactions with Pac1/LIS1 and Bik1/CLIP-170. Dynein is then delivered to the cell cortex and anchored through a cortical receptor protein, Num1. Num1 is a 313KDa protein that localizes to the cell cortex and is an essential component of dynein mediated nuclear migration.

Using quantitative fluorescence techniques I was able to create a molecular inventory of …

Sex Difference In Calbindin Cell Number In The Mouse Preoptic Area: Effects Of Neonatal Estradiol And Bax Gene Deletion, Richard F. Gilmore Iii

Masters Theses 1911 - February 2014

The sexually dimorphic nucleus of the preoptic area (SDN-POA) was first discovered in rats and is one of the most famous and best studied sex differences in the field of neuroscience. Though well documented in rats (larger in males than females), this sex difference was only recently able to be observed in mice due to the discovery of the protein calbindin-D28k as a marker. Recent studies have shown a larger, more distinct calbindin-immunoreactive (ir) cell cluster in male mice compared to females. However, the exact location of the cluster and whether the sex difference is one of total cell number …

Pharmacological Chaperoning In Fabry Disease, Jerome Rogich

Masters Theses 1911 - February 2014

Fabry Disease is an X-­‐linked lysosomal storage disorder characterized by a variety of symptoms including hypohydrosis, seizures, cardiac abnormalities, skin lesions, and chronic pain. These symptoms stem from a lack of functional endogenous α-­‐ Galactosidase A (α-­GAL), which leads to an accrual of its natural substrate. The severity of the disease symptoms can be directly correlated with the amount of residual enzyme activity. It has been shown that an imino sugar, 1-deoxygalactonojirimycin (DGJ), can increase enzymatic activity and clear excess substrate. This pH-­‐dependent chaperoning phenomenon is believed to arise from the presence of aspartic acid 170 in the active site. …

Novel Adaptor-Dependent Domains Promote Processive Degradation By Clpxp, Keith L. Rood

Masters Theses 1911 - February 2014

Protein degradation by ATP dependent proteases is a universally conserved process. Recognition of substrates by such proteases commonly occurs via direct interaction or with the aid of a regulatory adaptor protein. An example of this regulation is found in Caulobacter crescentus, where key regulatory proteins are proteolysed in a cell-cycle dependent fashion. Substrates include essential transcription factors, structural proteins, and second messenger metabolism components. In this study, we explore sequence and structural requirements for regulated adaptor mediated degradation of PdeA, an important regulator of cyclic-di-GMP levels.

Robust degradation of PdeA is dependent on the response regulator CpdR in vivo …

In Vivo Investigations Of Polymer Conjugates As Therapeutics, Elizabeth M. Henchey

Masters Theses 1911 - February 2014

Polymer conjugates offer a way to introduce materials into the body that would normally be rejected or cause toxicity. Two polymers are investigated in vivo for uses in chemotherapeutic delivery, protein therapeutics, and DNA transfection. A novel polymer, polyMPC, has the ability to increase doxorubicin loading and its solubility, and is conjugated in a way to release its payload in a low pH environment. Through its conjugation, blood clearance time of doxorubicin is increased, and thus tumor exposure to the drug is increased with a single administration. It can be administered at ten times the concentration of free doxorubicin, and …

Examination Of Promotor Hypermethylation Patterns In Magnetically Enriched Exfoliated Breast Milk Epithelial Cells, Chung M. Wong

Masters Theses 1911 - February 2014

Suppression of genes involved in DNA repair, tumor suppression and detoxification through epigenetic modifications has been implicated in the etiology of cancer. As such analysis of promoter methylation patterns in genes frequently down regulated in breast cancer in non-cancerous subjects may serve as an indicator of breast cancer risk. CpG-island hypermethylation of single genes has been detected in cells isolated from nipple aspirate and ductal lavage, yet both isolation methods yield insufficient cells to complete an extensive analysis on any one donor sample.

As an alternative we have turned to magnetic separation of human mammary epithelial cells from breast milk. …

Anti-Diabetic Potentials Of Phenolic Enriched Chilean Potato And Select Herbs Of Apiaceae And Lamiaceae Families, Fahad Saleem

Masters Theses 1911 - February 2014

The incidence of diabetes mellitus and cardiovascular diseases is increasing at a worrisome rate globally. Diabetes mellitus is known to occur due to high blood glucose levels, caused by defects in insulin levels. Adult on-set type II diabetes, which is closely associated with obesity, is reported to be 90-95% of all diabetic cases and linked to diet and lifestyle factors. A large population of the developed and developing countries is now being effected by this epidemic. Natural sources of phenolic antioxidants and inhibitors of digestive enzymes from food sources have potential for low cost dietary management of type II diabetes. …

In Vivo Labeling Of A Model Β-Clam Protein With A Fluorescent Amino Acid, Mangayarkarasi Periasamy

Masters Theses 1911 - February 2014

Proteins can be labeled with different tags to enable their structural and functional investigations. In addition, labeling proteins at specific sites helps in studying the conformational dynamics of these molecules. A plethora of methods is available to facilitate labeling, choice of which largely depends on the requirements and the anticipated end results. In general, the various labeling methods can be classified into four different classes based on the stage at which labeling is performed, namely post translational labeling, non-ribosomal synthesis, in vitro translation and in vivo translation. Interestingly all these techniques use different unnatural amino acids for this purpose.

Protein …

Dopamine Controls Locomotion By Modulating The Activity Of The Cholinergic Motor Neurons In C. Elegans, Andrew T. Allen

Masters Theses 1911 - February 2014

Dopamine is an important neurotransmitter in the brain, where it plays a regulatory role in the coordination of movement and cognition by acting through two classes of G protein-coupled receptors to modulate synaptic activity. In addition, it has been shown these two receptor classes can exhibit synergistic or antagonistic effects on neurotransmission. However, while the pharmacology of the mammalian dopamine receptors have been characterized in some detail, less is known about the molecular pathways that act downstream of the receptors. As in mammals, the soil nematode Caenorhabditis elegans uses two classes of dopamine receptors to control neural activity and thus …

Functional Analysis Of Receptor-Like Kinases In Pollen-Pistil Interactions In Arabidopsis Thaliana, Mini Aggarwal

Masters Theses 1911 - February 2014

Rac/Rop GTPases are molecular switches in plants that control the growth of polarized cells such as pollen tubes and root hairs, differentiation, development, actin dynamics, production of reactive oxygen species (ROS) and disease resistance. These small GTPases are activated by guanine nucleotide exchange factors (GEFs) that replace GDP for GTP and are referred to as RopGEFs in plants. To identify upstream components of the RopGEF regulated signaling pathways, GEF1 from Arabidopsis thaliana was used as a bait to screen a seedling cDNA library in a yeast two-hybrid system. This yielded members of a small family of the Catharanthus roseus Receptor-like …

Chemical Information Based Elastic Network Model: A Novel Way To Identification Of Vibration Frequencies In Proteins., Sharad K. Raj

Masters Theses 1911 - February 2014

A novel method of analysis of macromolecules has been worked upon through this research. In an effort to understand the dynamics of macromolecules and to further our knowledge, pertaining specifically to the low frequency domain and also to elucidate certain important biological functions associated with it, a theoretical technique of chemical information based Normal Mode Analysis has been developed. These simulations render users with the ability to generate animations of modeshapes as well as key insight on the associated vibration frequencies. Harmonic analysis using atomistic details is performed taking into account appropriate values of masses of constituent atoms of a …