Biochemistry, Biophysics, and Structural Biology Commons^™

Microsatellite Genotyping To Distinguish Colonies And Intraspecies Genetic Variation In The Eastern Subterranean Termite, Reticulitermes Flavipes (Isoptera: Rhinotermitidae), Timothy J. Husen, Shripat T. Kamble, Julie M. Stone

Department of Biochemistry: Faculty Publications

Microsatellite markers and fingerprinting techniques have demonstrated great potential for assessing colony affiliation in Reticulitermes species. This study utilized microsatellite allele fingerprints and examined variation at eight micro satellite loci within five different R. flavipes collection sites in each of Lancaster and Madison counties in Nebraska. In the termite populations from Lancaster County, five of the eight microsatellite loci (Rf 1-3, Rf 3-1, Rf 5-1 0, Rf 6-1, and Rf 24-2) provided informative genotypic variability allowing colony distinction. DNA sequence data at locus Rf24-2 also provided conclusive evidence that populations sampled were distinct colonies. Five micro satellite loci (Rf 3-1, …

Analysis Of The Mouse And Human Acyl-Coa Thioesterase (Acot) Gene Clusters Shows That Convergent, Functional Evolution Results In A Reduced Number Of Human Peroxisomal Acots., Mary Hunt, Anna Rautanen, Maria Westin, Thomas Svensson, Stefan Alexson

Articles

The maintenance of cellular levels of free fatty acids and acyl-CoAs, the activated form of free fatty acids, is extremely important as imbalances in lipid metabolism have serious consequences for human health. Acyl-CoA thioesterases (ACOTs) hydrolyze acyl-CoAs to the free fatty acid and CoASH, and thereby have the potential to regulate intracellular levels of these compounds. We have previously identified and characterized a mouse ACOT gene cluster, comprised of six genes that apparently arose by gene duplications, encoding acyl- CoA thioesterases with localizations in cytosol (ACOT1), mitochondria (ACOT2) and peroxisomes (ACOT3-6). However, the corresponding human gene cluster contains only three …

Rapid Cellular Signalling And Dose Dependent Cellular Pathways Induced In Hpv-G Cells Exposed To Medium Borne Factors, Paula Maguire

Doctoral

The aim of this thesis was to investigate rapid signalling events and dose dependent activation of cellular pathways in unirradiated human keratinocyte (HPV-G) cells exposed to bystander signal molecules in vivo and in vitro. HPV-G cells were exposed to irradiated cell conditioned medium (ICCM) ranging from 5mGy to 5Gy ICCM. Cellular events investigated included intercellular calcium, reactive oxygen species (ROS), mitochondrial membrane potential, cytochrome c release, caspase 3 and 8 activity, Bcl-2 expression and mitochondrial proliferation. Apoptosis levels and clonogenic survival were also determined in order to understand the cellular outcome of the activated pathways. It was found that exposure …

Dna Damage Responses In Progeroid Syndromes Arise From Defective Maturation Of Prelamin A, Michael Sinensky, Y. Liu, A. Rusinol, Y. Wang, Y. Zou

Faculty Publications, Biological Sciences

The genetic diseases Hutchinson-Gilford progeria syndrome (HGPS) and restrictive dermopathy (RD) arise from accumulation of farnesylated prelamin A because of defects in the lamin A maturation pathway. Both of these diseases exhibit symptoms that can be viewed as accelerated aging. The mechanism by which accumulation of farnesylated prelamin A leads to these accelerated aging phenotypes is not understood. Here we present evidence that in HGPS and RD fibroblasts, DNA damage checkpoints are persistently activated because of the compromise in genomic integrity. Inactivation of checkpoint kinases Ataxia-telangiectasia-mutated (ATM) and ATR (ATM- and Rad3-related) in these patient cells can partially overcome their …

Farnesylated Lamins, Progeroid Syndromes And Farnesyl Transferase Inhibitors, Michael Sinensky, A. E. Rusinol

Faculty Publications, Biological Sciences

Three mammalian nuclear lamin proteins, lamin B1, lamin B2 and the lamin A precursor, prelamin A, undergo canonical farnesylation and processing at CAAX motifs. In the case of prelamin A, there is an additional farnesylation-dependent endoproteolysis, which is defective in two congenital diseases: Hutchinson-Gilford progeria (HGPS) and restrictive dermopathy (RD). These two diseases arise respectively from defects in the prelamin A substrate and the enzyme (ZmpSte24) that processes it. Recent work has shed light on the roles of the lamin proteins and the enzymes involved in their farnesylation-dependent maturation. Other experimental work, including mouse model studies, have examined the possibility …

Monocyte Modulation Of Disease Pathogenesis And Progression In Localized Aggressive Periodontitis, Chu Ri Shin

Theses and Dissertations

Localized Aggressive Periodontitis (LAgP) is an aggressive, early onset form of periodontitis characterized by a unique myeloid cell phenotype. In addition to its bacterial origin, the unique phenotype of the myeloid cell contributes to disease pathogenesis and progression through mechanisms mediating host inflammatory and immune responses. LAgP monocytes synthesize increased levels of the potent proinflammatory lipid mediator, Prostaglandin E2 (PGE2), preferentially differentiate into dendritic cells, and lead to increased IgG2 production. In addition, levels of Platelet Activating Factor (PAF) have shown to be elevated in the gingival tissue and gingival crevicular fluid of subjects with periodontitis. The aim of this …

Molecular Epidemiology Of Antibiotic Resistant Salmonella Enterica Strains From Different Animal And Human Sources In Ireland, David George Lee

Theses

In this study, 176 isolates of Salmonella were collected from different laboratories in Ireland. All isolates were confirmed to be of the genus Salmonella by performing bacteriological and biochemical tests. Briefly, all isolates were cultured onto XLD agar after which pure cultures were verified and stocked at this institute. Definitive identification was carried out using the API 20-E system and a separate urease test was also performed on all isolates. After confirmation of the genus, all isolates were prepared for antibiotic susceptibility testing and r-types were determined using sixteen antibiotics. Depending on the antibiotic resistance profiles, isolates were selected for …

Characterizing An Rna Aptamer And Binding Studies With Synthetic Porphyrazines, Melanie Bozza

Seton Hall University Dissertations and Theses (ETDs)

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Development Of Single Nanoparticle Optical Assays For Imaging Single Living Cells, William John Brownlow

Chemistry & Biochemistry Theses & Dissertations

Multi-drug resistance (MDR) has been reported in both prokaryotes and eukaryotes; the pathogenic gram-negative bacteria Pseudomonas aeruginosa can extrude a variety of structurally and functionally diverse substrates via a number of membrane transport systems leading to MDR. We have developed a novel nanoparticle assay to characterize both the membrane transport system composed of the MexAB-OprM efflux pump and the membrane permeability induced by antibiotics. Gold (Au) and silver (Ag) nanoparticles were investigated for use as probes to explore membrane transport in P. aeruginosa.

The surface plasmon absorption (color) of Au nanoparticle solutions was found to change in the presence …

Exchange Coupling And Contribution Of Induced Orbital Angular Momentum Of Low-Spin Fe^3+ Ions To Magnetic Anisotropy In Cyanide-Bridged Fe2m2 Molecular Magnets: Spin-Polarized Density-Functional Calculations, Stephen Holmes, Kyungwha Park

Chemistry & Biochemistry Faculty Works

Electronic structure and intramolecular exchange constants are calculated for three cyanide-bridged molecular magnets, [Tp⋆FeIII(CN)3MII(DMF)4]2(OTf)2∙2DMF (MII=Mn,Co,Ni) (abbreviated as Fe2Mn2, Fe2Co2, and Fe2Ni2) that have been recently synthesized, within a generalized-gradient approximation in spin-polarized density-functional theory (DFT). Here Tp⋆=[C3(CH3)2HN2]3BH, OTf=O3SCF3, and DMF=HCON(CH3)2. Due to strong ligand fields present in the [Tp⋆FeIII(CN)3]− units, the Fe3+ ions exhibit a low ground-state spin of S=1/2. Our calculations show that the metal ions in the Fe2Mn2 molecule interact antiferromagnetically via cyanide ligands, while those in the Fe2Co2 and Fe2Ni2 molecule interact ferromagnetically. The calculations also suggest that the smallest gaps between the highest occupied molecular orbital …

Nutrient Control Of Microbial Carbon Cycling Along An Ombrotrophicminerotrophic Peatland Gradient, Jason K. Keller, Angela K. Bauers, Scott D. Bridgham, Laurie E. Kellogg, Colleen M. Iversen

Biology, Chemistry, and Environmental Sciences Faculty Articles and Research

Future climate change and other anthropogenic activities are likely to increase nutrient availability in many peatlands, and it is important to understand how these additional nutrients will influence peatland carbon cycling. We investigated the effects of nitrogen and phosphorus on aerobic CH4 oxidation, anaerobic carbon mineralization (as CO2 and CH4 production), and anaerobic nutrient mineralization in a bog, an intermediate fen, and a rich fen in the Upper Peninsula of Michigan. We utilized a 5-week laboratory nutrient amendment experiment in conjunction with a 6-year field nutrient fertilization experiment to consider how the relative response to nitrogen and phosphorus differed among …

Linking Ligand-Induced Alterations In Androgen Receptor Structure To Differential Gene Expression: A First Step In The Rational Design Of Selective Androgen Receptor Modulators, Dmitri Kazmin, Tatiana Prytkova, C. Edgar Cook, Russell Wolfinger, Tzu-Ming Chu, David Beratan, J. D. Norris, Ching-Yi Chang, Donald P. Mcdonnell

Biology, Chemistry, and Environmental Sciences Faculty Articles and Research

We have previously identified a family of novel androgen receptor (AR) ligands that, upon binding, enable AR to adopt structures distinct from that observed in the presence of canonical agonists. In this report, we describe the use of these compounds to establish a relationship between AR structure and biological activity with a view to defining a rational approach with which to identify useful selective AR modulators. To this end, we used combinatorial peptide phage display coupled with molecular dynamic structure analysis to identify the surfaces on AR that are exposed specifically in the presence of selected AR ligands. Subsequently, we …

Isolation And Functional Mapping Of Human T-Cell Leukemia Virus Type 1 Tax Oncoprotein Dna-Damage Complexes, Sarah Saionz Durkin

Theses and Dissertations in Biomedical Sciences

Human T-cell Leukemia Virus Type 1 (HTLV-1) is a transforming retrovirus which causes Adult T-cell Leukemia (ATL) and HTLV-1 Associated Myelopathy/Tropical Spastic Paraparesis (HAM/TSP). Cellular transformation can be caused by a single viral trans-activating protein, Tax. Tax may contribute to transformation through interaction with components of the DNA damage response pathway, promoting cellular genomic instability. We examined cellular Tax complexes in an effort to elucidate potential protein-protein interactions that can model the Tax-induced molecular events.

We also investigated the role of post-translational modification in regulating Tax function. We employed a direct physical analysis of Tax complexes isolated from mammalian …

Structure-Function Studies Of The Camp-Dependent Protein Kinase In Vitro And In Intact Cells, Gary Z. Morris

Theses and Dissertations in Biomedical Sciences

There are 518 protein kinase genes in the human genome; this constitutes about 1.7% of all human genes. The cAMP-dependent protein kinase (PKA) serves as the prototypic model for the study of kinases because it contains a conserved catalytic core shared with all eukaryotic kinases, it is the simplest kinase, and it is one of the best-characterized serine/threonine kinases. PKA is ubiquitous in mammals and regulates multiple physiological mechanisms such as the cell cycle, apoptosis, cell motility, energy metabolism, and gene transcription through a well-defined intracellular signaling pathway. While PKA clearly has a central physiological role it is still unclear …

The Use Of Proteomic Technologies To Identify Serum Glycoproteins For The Early Detection Of Liver And Prostate Cancers, Elizabeth Ellen Schwegler

Theses and Dissertations in Biomedical Sciences

The application of proteomic technologies to identify serum glycoproteins is an emerging technique to identify new biomarkers indicative of disease severity. Many of these newly evolving protein-profiling methodologies have evolved from previous global protein expression profiling studies such as those involving SELDI-TOF-MS technologies. Though the SELDI approach could distinguish disease from normal by utilizing protein patterns as shown herein with the HCC study of chapter II, it was unable to offer sequence information on the selected peaks, and did not have the ability to analyze the entire dynamic range of the serum/plasma proteome. To address these deficiencies, new strategies that …

Stretch-Dependant Tonic Force Maintenance In Rabbit Epigastric Artery, Krystina Michelle Berg

Theses and Dissertations

The contractile state of vascular smooth muscle (VSM) plays a key role in blood pressure regulation. Abnormal VSM contractility characterizes hypertension and understanding the regulatory mechanisms of VSM may provide some insights to specific treatment of hypertension. Upon muscle stimulation, Ca2+, myosin light chain (MLC) phosphorylation, crossbridge cycling rates and force increase to high levels, but with time, force is maintained while all other parameters of muscle activation fall to low levels. Thus, contraction is divided temporally into early (phasic) and late (tonic) phases, as determined by the underlying regulatory mechanism. Muscles with more phasic characteristics have a higher peak …

Inhibition Of Margination And Diapedesis Of Neutrophils By Protein Synthesis Blockade, Phyllis V. Acquah

Theses and Dissertations

Acute Respiratory Distress Syndrome (ARDS), an age-old clinical problem facing the Emergency Department and Intensive Care Units of all health systems, is a common debilitating lung condition consequent upon severe systemic inflammation. Although several studies have gone into understanding the epidemiology and pathogenesis of the disease thus making way for new advances in treatment strategies, there seems to be no known study tailored to its prevention. Neutrophil extravasation within the tissues during inflammation is the hallmark of this syndrome. Our study sought to block excessive neutrophil infiltration by inhibiting the biosynthesis of some essential proteins necessary for the process. In …

Characterization Of The Rank Ligand Positive Giant Cell Found In The Interfacial Membrane, Patrick Emerson Jones

Theses and Dissertations

Aseptic osteolysis is a major complication to total joint arthroplasty requiring several thousand people a year to have to undergo revisions of their joint prosthesis. The formation of the interfacial membrane has been associated with aseptic osteolysis leading to the failure of all types of total joints. Recent evidence suggests that RANKL, a potent activator of bone reabsorption, is present in the interfacial membrane. Prior research in this laboratory to determine the source of RANKL in the interfacial membrane has revealed the presence of intense areas of RANKL concentration in the membrane. These areas of RANKL concentration correspond to multiple …

Atm-Dependent Erk Signaling In Response To Dna Double Strand Breaks, Ashraf Khalil

Theses and Dissertations

Ionizing radiation (IR) triggers many signaling pathways stemming from DNA damage, and, independently, from extra-nuclear events. To generate radio-mimetic DNA double-strand breaks (DSBs) without and minimizing the effects on extra-nuclear radiation targets, human (p53+) glioma and carcinoma cells containing bromodeoxyuridine (BrdU)- substituted DNA were treated with Hoechst 33258 followed by long wave-length UV (UV-A) (BrdU photolysis). BrdU photolysis resulted in well-controlled, dose-dependent generation of DSBs equivalent to 0.2 - 20 Gy of IR, as detected by pulse-field gel electrophoresis, accompanied by dose-dependent H2AX phosphorylation at ser-139 and ATM phosphorylation at ser-1981, indicating ATM activation. Furthermore, BrdU photolysis increased phosphorylation of …

Phospholipase A2 Induced Monocyte Chemotaxis To Apoptotic Cells, Kwasi Karikari

Theses and Dissertations

Apoptosis is a form of programmed cell death that is essential in such processes as organ and tissue remodeling and maturation of hematopoietic cells. The clearance of apoptotic cells is essential to prevent autoimmune responses to sequestered antigens. This process is mediated by phagocytes of the monocyte lineage. Before phagocytosis can occur, macrophages must be recruited to the apoptotic cells through chemotaxis. Products of the reaction catalyzed by the phospholipases A2 (PLA2) have been shown to induce monocyte chemotaxis either directly or indirectly. Some investigators have implicated a cytosolic calcium-independent PLA2 (iPLA2) in the production of these products during apoptosis. …

Characterization Of The Ftf/Hnf-4 Sites Within The 7alpha- And The 12alpha-Hydroxylase Promoters Involved In The Bile Acid-Mediated Transcription Of Their Regulation, Preeti Pramanik

Theses and Dissertations

Bile acids regulate their own synthesis through a feedback regulatory mechanism of mainly two enzymes in the classic pathway, the 7α-hydroxylase and the 12α-hydroxylase. In the early 1990's it was shown that the regulatory responses of 7α-hydroxylase are mediated at the transcriptional level and since then many positive and negative transcription factors that mediate regulatory response have been identified. An important finding was that the transcription factors regulating the expression of 7α- and 12α-hydroxylase genes are nuclear receptors.One of the first nuclear receptors identified to play a role in the transcription of the 7α-hydroxylase gene was HNF-4 since then many …

I. Comparison Of Translesion Bypass Of Guanine–N2 Monoadducts Of Mitomycin C And Guanine-N7 Monoadducts Of 2,7-Diaminomitosene By T7 Exo-, Klenow Exo-, Eta And Klenow Exo+ Dna Polymerases. Ii. Structure-Based Design, Synthesis, Structure-Conformation And Structure-Activity Relationships Studies Of D-Phe-Pro-D-Arg-P1’-Conh2 Tetrapeptides With Inhibitory Activity For Thrombin., Cristina C. Clement

Dissertations, Theses, and Capstone Projects

The guanine (G)-N² DNA monoadduct of mitomycin C (MC), a cytotoxic anticancer drug, inhibits translesion bypass by DNA polymerases. 2,7-Diaminomitosene (2,7-DAM) is the major metabolite of MC in tumor cells, generated by the reduction of MC. 2,7-DAM alkylates DNA in the cell in situ, forming an adduct at the N7 position of 2'-deoxyguanosine (2,7-DAM-dG) and is noncytotoxic. In part I of this study we tested a potential correlation between the lack of cytotoxicity of 2,7-DAM and the relative ease of bypass of this adduct as compared with the MC adduct. 24-mer and 27-mer templates, adducted at a single …