Molecular Biology Commons^™

A Novel Function For Aurora B Kinase In The Regulation Of P53 By Phosphorylation, Chris P. Gully

Dissertations and Theses (Open Access)

The mitotic kinase Aurora B plays a pivotal role in mitosis and cytokinesis and governs the spindle assembly checkpoint which ensures correct chromosome segregation and normal progression through mitosis.  Aurora B is overexpressed in breast and other cancers and may be an important molecular target for chemotherapy.  Tumor suppressor p53 is the guardian of the genome and an important negative regulator of the cell cycle. Previously, it was unknown whether Aurora B and p53 had mutual regulation during the cell cycle.  A small molecule specific inhibitor of Aurora B, AZD1152, gave us an indication that Aurora B negatively impacted p53 …

Regulation Of Set1-Mediated Methylation Of Dam1, John A. Latham

Dissertations and Theses (Open Access)

Eukaryotic genomes exist within a dynamic structure named chromatin in which DNA is wrapped around an octamer of histones forming the nucleosome. Histones are modified by a range of posttranslational modifications including methylation, phosphorylation, and ubiquitination, which are integral to a range of DNA-templated processes including transcriptional regulation. A hallmark for transcriptional activity is methylation of histone H3 on lysine (K) 4 within active gene promoters. In S. cerevisiae, H3K4 methylation is mediated by Set1 within the COMPASS complex. Methylation requires prior ubiquitination of histone H2BK123 by the E2-E3 ligases Rad6 and Bre1, as well as the Paf1 transcriptional …

Expression And Regulation Of Human Cytochrome P450 4f Isoforms In Tissue Samples And Under Tnf-Alpha Challenges, Jordan C. Bell

Dissertations and Theses (Open Access)

CYP4F (Cytochrome P4504F) enzymes metabolize endogenous molecules including leukotrienes, prostaglandins and arachidonic acid. The involvement of these endogenous compounds in inflammation has led to the hypothesis that changes in the inflamed tissue environment may affect the expression of CYP4Fs during the pro-inflammatory state, which in turn may modulate inflammatory conditions during the anti-inflammatory state. We demonstrated that inflamed tissues have different levels of CYP4F isoform expression profiles in a number of human samples when compared to the average population. The CYP4F isoform expression levels change with the degree of inflammation present in tissue. Further investigation in cell culture studies revealed …

Ancient Mitochondrial Dna From Pre-Historic Southeastern Europe: The Presence Of East Eurasian Haplogroups Provides Evidence Of Interactions With South Siberians Across The Central Asian Steppe Belt, Jeremy R. Newton

Masters Theses

Studies of mitochondrial DNA (mtDNA) polymorphism have provided valuable insights for understanding patterns of human migration and interaction. The ability to recover ancient mtDNA sequence data from post-mortem bone and tissue samples allows us to view snapshots of historic gene pools firsthand, provided that great care is taken to prevent sample contamination. In this study, we analyzed the DNA sequence of the first hypervariable segment (HVSI) of the mtDNA control region, as well as a portion of the coding region, in 14 individuals from three collective burials from the Neolithic Dnieper-Donetz culture and three individuals from Bronze Age Kurgan burials, …

Functional Studies Of Human Cellular Detoxification Enzymes, Melanie Neely Willis

Department of Biochemistry: Dissertations, Theses, and Student Research

Cellular detoxification allows for the maintenance of cellular homeostasis and prevention of abnormal cell growth by clearing harmful xenobiotics and endobiotics. After oxygenation by phase I enzymes, phase II enzymes such as glucuronosyltransferases and glutathione-s-transferases conjugate a small molecule to the compound, marking it for subsequent export. Many up-stream enzymes are also essential to cellular detoxification by supplying the small compounds for conjugation. These up-stream enzymes include UDP-glucose dehydrogenase, which synthesizes UDP-glucuronate, and glutamate cysteine ligase, which catalyzes the first and rate-limiting step in the synthesis of glutathione.
UDP-glucose dehydrogenase (UGDH) is an important enzyme in human development and in …

The Release Of Calcium In Bacillus Anthracis Pathogenicity Methods, Natiera Magnuson, Manomita Patra Bhowmik, Maria Elena Reynaga, Ernesto Abel-Santos

Festival of Communities: UG Symposium (Posters)

Anthrax infection starts with germination of Bacillus anthracis spores in macrophages. Some bacteria, including B. anthracis, can sporulate in response to environmental stress, such as starvation. During germination, large concentrations of calcium ions are released from the B. anthracis spore. Calcium ions are hydrophilic secondary messengers, and may therefore interfere with detection of the spore by confusing the cell signaling pathways. We investigated calcium release on infected macrophage viability by replacing the calcium stored in B. anthracis spores for other cations via demineralization/remineralization. It was discovered that calcium ions typically out-performed other cations in germination of B. anthracis.

Characterization Of Ips Pini Ipsdienol Dehydrogenase (Idol Dh), Heidi Pak, Claus Tittiger, William Welch

Festival of Communities: UG Symposium (Posters)

Ipsdienol is an important pheromone component for pine engraver beetle, Ips pini. Ipsdienol is a ten carbon monoterpenoid secondary alcohol and ipsdienone is the corresponding ketone. We are characterizing the activity of recombinant IDOL DH produced in Sf9 (insect) cells. The enzyme has a high stereospecificity: (-) ipsdienol was found to be a substrate while (+)-ipsdienol was neither a substrate nor inhibitor. Closely related monoterpenoids, such as nerol, geraniol, and citral, were neither substrates nor inhibitors. Smaller compounds, such as 2-propanol, also failed to act as an inhibitor or substrate. This indicates the binding site of this enzyme is highly …

Arctic Mustard Flower Color Polymorphism Controlled By Petal-Specific Downregulation At The Threshold Of The Anthocyanin Biosynthetic Pathway, Cynthia A. Dick, Jason Buenrostro, Timothy Butler, Matthew L. Carlson, Daniel J. Kliebenstein, Justen B. Whittall

Biology

Intra- and interspecific variation in flower color is a hallmark of angiosperm diversity. The evolutionary forces underlying the variety of flower colors can be nearly as diverse as the colors themselves. In addition to pollinator preferences, non-pollinator agents of selection can have a major influence on the evolution of flower color polymorphisms, especially when the pigments in question are also expressed in vegetative tissues. In such cases, identifying the target(s) of selection starts with determining the biochemical and molecular basis for the flower color variation and examining any pleiotropic effects manifested in vegetative tissues. Herein, we describe a widespread purple-white …

Constructing A Vector For Over Expressing Nitrobenzene Dioxygenase In E. Coli, Paul Davis

Honors Student Works

Due to extensive use of nitroaromatic compounds in the production of dyes, pesticides, explosives, and their harmful effects on the environment, studying the enzyme mechanisms of bacteria capable of utilizing these compounds as nutrient sources have become a recent field of development. One example, the Comamonas sp. strain JS765, grows on nitrobenzene as its sole carbon, nitrogen and energy source. This bacterium contains a nitrobenzene dioxygenase enzyme system that incorporates O2 into the nitrobenzene ring to form catechol and nitrite. In this project, the genes that code for the dioxygenase enzyme were cloned and then transformed into an expression vector …

Section Abstracts: Biology With Microbiology And Molecular Biology

Virginia Journal of Science

Abstracts of the Biology with Microbiology and Molecular Biology Section for the 89th Annual Meeting of the Virginia Academy of Science, May 25-27, 2011, University of Richmond, Richmond VA.

An Observation Of Immunological Effect, A Diet Enhanced With Spirulina And Treatment With Fractalkine In Models Of Parkinson's Disease, Mibel M. Pabón

USF Tampa Graduate Theses and Dissertations

In my dissertation research we used use human wild type α-synuclein gene expression using an adeno-associated viral vector (AAV9) that induced a slowly progressive loss of dopamine (DA) neurons in the Substantia nigra (SN) as one of our animal model of Parkinson’s disease (PD). It is our hypothesis that neuroinflammation predisposes the brain to susceptibility to neurodegenerative diseases. Thus we examined the progression of a PD lesion and examined the manipulations of the immune system to understand further the inflammatory role when we administered exogenous soluble fractalkine.

The specific etiology of neurodegeneration in PD is unknown, but the inflammatory mechanisms …

Structural Study Of Mcry2: A Circadian Rhythm Regulator, Brenda L. Rodarte^, Gustavo A. Avila, Adrian S. Enriquez, Nicolas D. Silva, Tiffany H. Blankenship, Erquan Eric Zhang*, Chuan Xiao*

COURI Symposium Abstracts, Spring 2011

A circadian clock is a 24 hour biological rhythm that regulates physiological, psychological and behavioral processes in living organisms. Understanding the regulation mechanism of such circadian rhythms can have great impact in certain diseases such as sleeping disorders and cancers. There are many genes regulating circadian rhythms in different species. Cryptochromes (CRY) are photosensory receptors mediating light regulation of growth and development in certain species. Mouse cryptochrome protein mCRY2 has been found to encode a blue light photoreceptor and is important for mice circadian rhythm. The main goal of the project is to determine the atomic structure of mCRY2, via …

Scireader Enables Reading Of Medical Content With Instantaneous Definitions, Patrick R. Gradie, Megan Litster, Rinu Thomas, Jay Vyas, Martin Schiller

Life Sciences Faculty Research

Background

A major problem patients encounter when reading about health related issues is document interpretation, which limits reading comprehension and therefore negatively impacts health care. Currently, searching for medical definitions from an external source is time consuming, distracting, and negatively impacts reading comprehension and memory of the material.

Methods

SciReader was built as a Java application with a Flex-based front-end client. The dictionary used bySciReader was built by consolidating data from several sources and generating new definitions with a standardized syntax. The application was evaluated by measuring the percentage of words defined in different documents. A survey was used …

Unsupplemented Artemia Diet Results In Reduced Growth And Jaw Dysmorphogenesis In Zebrafish, Michael P. Craig, Mitul B. Desai, Kate E. Olukalns, Scott E. Afton, Joseph A. Caruso, Jay R. Hove

Biochemistry and Molecular Biology Faculty Publications

The number of laboratories using zebrafish as an experimental animal model has risen tremendously over the past two decades (Craig et al., 2006). As a result, the number of zebrafish facilities around the world has dramatically increased to meet the elevated demand for proper animal care and maintenance. In order to meet this demand, aquaculture facilities must employ husbandry protocols designed to produce a constant supply of healthy, viable eggs. Surprisingly, many husbandry strategies, particularly feeding protocols, are frequently passed down from members of one lab to another in a colloquial fashion without rigorous experimental validation. An ideal diet should …

Inhibition Of Proprotein Convertase Ski-1 Blocks Transcription Of Key Extracellular Matrix Genes Regulating Osteoblastic Mineralization, Jeff P. Gorski, Nichole T. Huffman, Sridar Chittur, Ronald J. Midura, Claudine Black, Julia Oxford, Nabil G. Seidah

Biomedical Research Institute Publications and Presentations

Mineralization, a characteristic phenotypic property of osteoblastic lineage cells, was blocked by AEBSF and dec-RRLL-cmk, inhibitors of SKI-1 (site 1; subtilisin kexin like-1) protease. Since SKI-1 is required for activation of SREBP and CREB/ATF family transcription factors, we tested the effect of these inhibitors on gene expression. AEBSF decreased expression of 140 genes by 1.5- to 3.0-fold including Phex, Dmp1, COL1A1, COL11A1 and fibronectin. Direct comparison of AEBSF and dec-RRLL-cmk, a more specific SKI-1 inhibitor, demonstrated that expression of Phex, Dmp1, COL11A1 and fibronectin was reduced by both while COL1A2 and HMGCS1 were reduced …

Dynamic Glucoregulation And Mammalian-Like Responses To Metabolic And Developmental Disruption In Zebrafish, Agata Jurczyk, Nicole M. Roy, Rabia Bajwa, Philipp Gut, Kathryn Lipson, Chaoxing Yang, Laurence Covassin, Waldemar J. Racki, Aldo A. Rossini, Nancy Phillips, Didier Y. R. Stainier, Dale L. Greiner, Michael A. Brehm, Rita Bortell, Philip Diiorio

Biology Faculty Publications

Zebrafish embryos are emerging as models of glucose metabolism. However, patterns of endogenous glucose levels, and the role of the islet in glucoregulation, are unknown. We measured absolute glucose levels in zebrafish and mouse embryos, and demonstrate similar, dynamic glucose fluctuations in both species. Further, we show that chemical and genetic perturbations elicit mammalian-like glycemic responses in zebrafish embryos. We show that glucose is undetectable in early zebrafish and mouse embryos, but increases in parallel with pancreatic islet formation in both species. In zebrafish, increasing glucose is associated with activation of gluconeogenic phosphoenolpyruvate carboxykinase1 (pck1) transcription. Non-hepatic Pck1 protein is …

Novel Copper Loaded Core-Shell Silica Nanoparticles With Improved Copper Bioavailability Synthesis, Characterization And Study Of Antibacterial Properties, Pavithra Maniprasad

Electronic Theses and Dissertations

A novel core-shell silica based antimicrobial nanoparticle was synthesized. The Stöber silica shell has been engineered to accommodate copper. Synthesis of the core-shell Cu-silica nanoparticle (C-S CuSiNP) involves preparation of base-hydrolyzed Stöber silica “seed” particles first, followed by the acid-catalyzed seeded growth of the Cu-silica shell layer around the core. Scanning electron microscopy and transmission electron microscopy showed monodispersed, spherical shaped nanoparticles with smooth surface morphology. Characterization of particle size distribution in solution by the Dynamic Light Scattering (DLS) technique was fairly consistent with the electron microscopy results. Loading of Cu to nanoparticles was confirmed by the SEM-Energy Dispersive X-Ray …

A Solid Phase Assay For Topoisomerase I Interfacial Poisons And Catalytic Inhibitors, Vidusha Cyril

Electronic Theses and Dissertations

We report a mechanism based screening technique to rapidly identify eukaryotic topoisomerase I targeting agents. The method is based on genetic tagging of topoisomerase I to immobilize the enzyme on a solid surface in a microtiter well format. DNA is added to the wells and retained DNA is detected by Picogreen fluorescence. Compounds that result in an increase in Picogreen staining represent potential topoisomerase interfacial poisons while those that reduce fluorescence report catalytic inhibitors; therefore, the solid phase assay represents a „bimodal‟ readout that reveals mechanisms of action. The method has been demonstrated to work with known interfacial poisons and …

Performance Screening Of Chemostat Adapted Recombinant Zymomonas Mobilis Strains, Deirdre M. Beard, Nancy Dowe

STAR Program Research Presentations

Corn stover biomass can be pretreaed and hydrolyzed into soluble sugars to be fermented by microorganisms to ethanol. NREL has developed a recombinant bacteria Zymomonas mobilis 8b that metabolizes both five and six carbon sugars. During pretreatment, toxic inhibitors such as furfural and acetate are produced. NREL has made an attempt to adapt two sub-strains of Z. mobilis 8b to acetate and furfural by using a chemostat method. During the chemostat process, cultures were frozen back in glycerol and saved. In this study, those frozen cultures were revived and analyzed for performance in environments with varying concentrations of furfural and …

The Prevalence Of Canine Leishmania Infantum Infection In Western China Detected By Pcr And Serological Tests, Jun-Yun Wang, Yu Ha, Chun-Hua Gao, Yong Wang, Yue-Tao Yang, Hai-Tang Chen

Wayne State University Associated BioMed Central Scholarship

Abstract

Background

Canine leishmaniasis (CanL) is endemic in western China, resulting in important public health problem. It is essential to evaluate the prevalence of canine Leishmania infantum infection for designing control policy. In the present study we report for the first time prevalence of Leishmania infection in dogs living in Jiuzhaigou County (Sichuan Provence, China), which is not only an important endemic area of CanL but also a tourism scenic spot, detected by PCR, ELISA and dipstick test. The results could provide key information for designing control programs against canine and human leishmaniasis. In addition, the complete sequence of the …