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Articles 1 - 14 of 14

Full-Text Articles in Molecular Biology

Characterizing The Human Vaginal Microbiome Using High-Throughput Sequencing, Jean Megan E. Macklaim Dec 2013

Characterizing The Human Vaginal Microbiome Using High-Throughput Sequencing, Jean Megan E. Macklaim

Electronic Thesis and Dissertation Repository

The human vaginal microbiome undoubtedly has a significant role in reproductive health and for protection from infectious organisms. Recent efforts to characterize the bacterial species of the vagina using molecular techniques have uncovered an unexpected diversity. Using high-throughput sequencing I sought to describe the structure and function of the vaginal microbiome under different physiological states including healthy, bacterial vaginosis (BV), post-menopausal vaginal atrophy, and acute vulvovaginal candidiasis (VVC).

Partial 16S rRNA gene sequencing revealed that healthy, asymptomatic women most often have vaginal biotas dominated by Lactobacillus iners or L. crispatus. In contrast, BV is a heterogeneous, highly diversified condition with ...


Exploring The Structure And Biochemistry Of Oxidation-Mediated Inhibitation Of The Peptidyl-Prolyl Isomerase Pin1, Brendan T. Innes Dec 2013

Exploring The Structure And Biochemistry Of Oxidation-Mediated Inhibitation Of The Peptidyl-Prolyl Isomerase Pin1, Brendan T. Innes

Electronic Thesis and Dissertation Repository

Pin1 is a phosphorylation-dependent peptidyl-prolyl isomerase that has been shown to be neuroprotective in aging-related neurodegenerative diseases such as Alzheimer's disease (AD). However, it is not active in AD brain, and a recent proteomic screen of Mild Cognitive Impairment (MCI) brain samples revealed that Pin1 is oxidized in the brains of these pre-AD patients. This suggests that this oxidation may be the cause of the loss of the neuroprotective Pin1 function in AD. The Pin1 active site contains a functionally critical cysteine residue (Cys113) with a low predicted pKa, making it highly susceptible to oxidation. We hypothesize that Pin1 ...


Uncovering The Molecular Link Between Mir156.Spl15 And Carotenoid Accumulation In Arabidopsis, Davood Emami Meybodi Oct 2013

Uncovering The Molecular Link Between Mir156.Spl15 And Carotenoid Accumulation In Arabidopsis, Davood Emami Meybodi

Electronic Thesis and Dissertation Repository

Carotenoid Cleavage Dioxygenases (CCDs) are an enzyme family that cleaves specific double bonds in carotenoids. MicroR156 in Arabidopsis regulates a network of genes by repressing 10 SPL genes, among which, SPL15 was found to regulate shoot branching and carotenoid accumulation. The expression of CCD1, CCD4, CCD7, CCD8, NCED2, NCED3, NCED5, NCED6, NCED9 and SPL15 was evaluated in siliques at 10 days post anthesis and in 10-day-old roots in Arabidopsis wild type, sk156 (miR156 overexpression mutant), RS105 (miR156 overexpression line), spl15 (SPL15 knockout mutant) and two 35S:SPL15 lines. Results showed that most of CCD/NCED genes were affected ...


Fabricating Cost-Effective Nanostructures For Biomedical Applications, Erden Ertorer Sep 2013

Fabricating Cost-Effective Nanostructures For Biomedical Applications, Erden Ertorer

Electronic Thesis and Dissertation Repository

In this thesis we described inexpensive alternatives to fabricate nanostructures on planar substrates and provided example applications to discuss the efficiency of fabricated nanostructures.

The first method we described is forming large area systematically changing multi-shape nanoscale structures on a chip by laser interference lithography. We analyzed the fabricated structures at different substrate positions with respect to exposure time, exposure angle and associated light intensity profile. We presented experimental details related to the fabrication of symmetric and biaxial periodic nanostructures on photoresist, silicon surfaces, and ion-milled glass substrates. Behavior of osteoblasts and osteoclasts on the nanostructures was investigated. These results ...


Absence Of Phosphoenolpyruvate Carboxylase Atppc3 Increases Sensitivity Of Arabidopsis Thaliana To Cadmium, Ian R. Willick Jul 2013

Absence Of Phosphoenolpyruvate Carboxylase Atppc3 Increases Sensitivity Of Arabidopsis Thaliana To Cadmium, Ian R. Willick

Electronic Thesis and Dissertation Repository

Phosphoenolpyruvate carboxylase (PEPC) and PEPC kinase (PPCK) catalyze a reaction feeding into the tricarboxylic acid (TCA) cycle, increasing the production of metal-chelating organic acids. Little research has been conducted on PEPC isoenzymes in Cd-stressed plants. Arabidopsis (Arabidopsis thaliana) wild-type and AtPPC1 – AtPPC3 mutants, each lacking one of three PEPC isoenzymes, grown in 0, 1, or 5 µM CdCl2 were smaller and had increased AtPPC1 – AtPPC3 and AtPPCK1 – AtPPCK2 transcript abundance, relative phosphorylation, and PEPC activity, more so in roots than shoots. Concentrations of oxaloacetate, citrate and total organic acids increased with greater CdCl2 concentrations. The absence of AtPPC3 ...


Molecular Mechanisms Underlying The Early Life Programming Of The Liver, Gurjeev Sohi Jul 2013

Molecular Mechanisms Underlying The Early Life Programming Of The Liver, Gurjeev Sohi

Electronic Thesis and Dissertation Repository

Clinical studies have demonstrated that intrauterine growth restriction (IUGR) offspring, faced with a nutritional mismatch postpartum, have an increased risk of developing the metabolic syndrome. The maternal protein restriction (MPR) rat model has been extensively studied to investigate the adverse effects of a nutritional mismatch in postnatal life of IUGR offspring. Previous studies have demonstrated that MPR leads to impaired function of the liver, an important metabolic organ. However the underlying mechanisms which predispose these offspring to the metabolic syndrome remain elusive. In the following studies, low protein diet during pregnancy and lactation led to IUGR offspring with decreased liver ...


Identification Of Cyclophilin Gene Family In Soybean And Characterization Of Gmcyp1, Hemanta Raj Mainali Jul 2013

Identification Of Cyclophilin Gene Family In Soybean And Characterization Of Gmcyp1, Hemanta Raj Mainali

Electronic Thesis and Dissertation Repository

I identified members of the Cyclophilin (CYP) gene family in soybean (Glycine max) and characterized the GmCYP1, one of the members of soybean CYP. CYPs belong to the immunophilin superfamily with peptidyl-prolyl cis-trans isomerase (PPIase) activity. PPIase catalyzes the interconversion of the cis- and trans-rotamers of the peptidyl-prolyl amide bond of peptides. After extensive data mining, I identified 62 different CYP genes in soybean (GmCYP1 to GmCYP62), of which 8 are multi-domain proteins and 54 are single domain proteins. At least 25% of the GmCYP genes are expressed in soybean. GmCYP1 localizes to the nucleus and the cytoplasm and ...


Human Adenovirus E1a Binds And Retasks Cellular Hbre1, Blocking Interferon Signalling And Activating Virus Early Gene Transcription, Gregory J. Fonseca Jun 2013

Human Adenovirus E1a Binds And Retasks Cellular Hbre1, Blocking Interferon Signalling And Activating Virus Early Gene Transcription, Gregory J. Fonseca

Electronic Thesis and Dissertation Repository

Upon infection, human adenovirus (HAdV) must block interferon signaling and activate the expression of its early genes to reprogram the cellular environment to support virus replication. During the initial phase of infection, these processes are orchestrated by the first HAdV gene expressed during infection, early region 1A (E1A). E1A binds and appropriates components of the cellular transcriptional machinery to modulate cellular gene transcription and activate viral early genes transcription. We have identified hBre1/RNF20 as a novel target of E1A. hBre1 is an E3 ubiquitin ligase which acts with the Ube2b E2 conjugase and accessory factors RNF40 and WAC1 to ...


Regulation Of Lipid Homeostasis, Inflammatory Signalling And Atherosclerosis By The Peroxisome Proliferator-Activated Receptor Delta, Lazar A. Bojic Jun 2013

Regulation Of Lipid Homeostasis, Inflammatory Signalling And Atherosclerosis By The Peroxisome Proliferator-Activated Receptor Delta, Lazar A. Bojic

Electronic Thesis and Dissertation Repository

The peroxisome proliferator-activated receptor (PPAR) δ is a ligand-dependent transcription factor that has been implicated in metabolic and inflammatory regulation. The molecular and physiological mechanisms by which PPARδ activation regulates lipid metabolism, inflammatory signaling and protection from atherosclerosis in states of metabolic disturbance such as insulin resistance and dyslipidemia, were investigated in a series of in vitro and in vivo studies. In vitro experiments demonstrated that PPARδ activation inhibits atherogenic lipoprotein-induced lipid accumulation and the associated proinflammatory responses. The primary mechanisms for these effects were increased fatty acid β-oxidation, decreased lipoprotein lipase (LPL) activity, reduced MAPK signaling and improved insulin ...


Metabolism And Transcriptional Responses To Asparagine In Arabidopsis Thaliana, Qianyi Zhang Apr 2013

Metabolism And Transcriptional Responses To Asparagine In Arabidopsis Thaliana, Qianyi Zhang

Electronic Thesis and Dissertation Repository

Asparagine aminotransferase transforms asparagine into α-ketosuccinamate, which is further deamidated by an ω-amidase. Serine:glyoxylate aminotransferase, encoded by AGT1 in Arabidopsis, was identified as asparagine aminotransferase. In the roots of 10-day-old Arabidopsis seedlings treated with 20 mM asparagine, AGT1 transcript levels increased by 2-fold while ω-amidase transcripts were decreased by 30%. Recombinant AGT1 had a substrate preference for asparagine when compared with alanine and serine as amino group donors. An ω-amidase candidate gene, AT5G12040, was identified based on amino acid sequence identity with mammalian gene Nitrilase 2. RT-PCR of a T-DNA insertion mutant line showed that ω-amidase expression was abolished ...


Cellular Adaptation Of Macrophages To Anthrax Lethal Toxin-Induced Pyroptosis Via Epigenetic Mechanisms, Chae Young Han Apr 2013

Cellular Adaptation Of Macrophages To Anthrax Lethal Toxin-Induced Pyroptosis Via Epigenetic Mechanisms, Chae Young Han

Electronic Thesis and Dissertation Repository

Cellular adaptation to microbial stresses has been demonstrated in several cell types. Macrophages (MФ) are sentinel immune cells fending off invading microbes. Anthrax lethal toxin (LeTx) is a key virulence factor released by Bacillus anthracis that causes rapid cell death, pyroptosis. A small number of RAW246.7 macrophages (~4%) exposed to a non-lethal dose of LeTx become resistant to LeTx-induced pyroptosis for ~ 4 weeks, termed “toxin-induced resistance (TIR)”. Here, I showed that high levels of DNA methyl transferase1 (DNMT1) expression were maintained although global genomic methylation levels were not high in TIR. TIR cells treated with the DNMT inhibitor 5-azacitidine ...


Dna Hydrolysis And Genome Editing Applications Of Giy-Yig Family Homing Endonucleases, Benjamin P. Kleinstiver Apr 2013

Dna Hydrolysis And Genome Editing Applications Of Giy-Yig Family Homing Endonucleases, Benjamin P. Kleinstiver

Electronic Thesis and Dissertation Repository

The ability to manipulate complex genomes in a precise manner is essential for studying biological processes in model systems, engineering plant strains for agriculture, or advancing human cellular therapies to treat diseases. Genomic alterations are most efficient when a double-strand DNA break is introduced at the loci where the modification is desired. Different classes of naturally occurring DNA endonucleases, including homing endonucleases, have therefore been explored as candidates for genome modification studies as they target long stretches of DNA. Homing endonucleases are mobile genetic elements whose biological role is to introduce site-specific double-strand breaks into naïve genomes, ultimately resulting in ...


Transcriptional Regulation By The Oncogenic Znf217/Corest Complex, Gobi Thillainadesan Apr 2013

Transcriptional Regulation By The Oncogenic Znf217/Corest Complex, Gobi Thillainadesan

Electronic Thesis and Dissertation Repository

The ZNF217 transcription factor is an oncogene found within the 20q13 amplicon and is amplified and overexpressed in many cancers including breast and ovarian. Overexpression of ZNF217 leads to increased cell proliferation, survival, and causes resistance to TGFβ's anti-proliferative effects.

ZNF217 is a core constituent of a transcriptional complex that includes CoREST, HDAC1/2, LSD1, and the CtBP1/2. In this study, I have combined genome-wide biochemical approaches to identify genes directly regulated by ZNF217. I have identified the tumor suppressor and cell cycle inhibitor, p15ink4b, as a direct target of the ZNF217 complex and demonstrated that ZNF217 ...


The Effect Of Glycan Linked To Cationic Peanut Peroxidase On Its Activity, Maha M. Al-Subaie Mar 2013

The Effect Of Glycan Linked To Cationic Peanut Peroxidase On Its Activity, Maha M. Al-Subaie

Electronic Thesis and Dissertation Repository

The roles of heme-, calcium- and glycan moieties on the peanut peroxidase enzyme activity have been previously studied. While the 3-D protein structure is known and some information about the glycan chain is known, due to uneven distribution of the 5 glycans, only a partial understanding of the furthest sugar groups on the 16 glycan chain was available. Now, having used gel-filtration column chromatography followed by lectin affinity column work, I showed that there is a rich array of galactose there. Moreover, I determined that not too many of the sugars are removed in the culture period of 14 days ...