Biochemistry, Biophysics, and Structural Biology Commons^™

Low-Ω3 Fatty Acid And Soy Protein Attenuate Alcohol-Induced Fatty Liver And Injury By Regulating The Opposing Lipid Oxidation And Lipogenic Signaling Pathways., Karina Reyes-Gordillo, Ruchi Shah, Ravi Varatharajalu, Mamatha Garige, Leslie C. Leckey, M. Raj Lakshman

Biochemistry and Molecular Medicine Faculty Publications

Chronic ethanol-induced downregulation of peroxisome proliferator-activated receptor gamma coactivator 1-alpha (PGC1α) and upregulation of peroxisome proliferator-activated receptor gamma coactivator 1-beta (PGC1β) affect hepatic lipid oxidation and lipogenesis, respectively, leading to fatty liver injury. Low-ω3 fatty acid (Low-ω3FA) that primarily regulates PGC1α and soy protein (SP) that seems to have its major regulatory effect on PGC1β were evaluated for their protective effects against ethanol-induced hepatosteatosis in rats fed with Lieber-deCarli control or ethanol liquid diets with high or low ω3FA fish oil and soy protein. Low-ω3FA and SP opposed the actions of chronic ethanol by reducing serum and liver lipids with …

And-1 Is Required For Homologous Recombination Repair By Regulating Dna End Resection, Yongming Li, Zongzhu Li, Zhiyong Han, Wenge Zhu

Biochemistry and Molecular Medicine Faculty Publications

Homologous recombination (HR) is a major mechanism to repair DNA double-strand breaks (DSBs). Although tumor suppressor CtIP is critical for DSB end resection, a key initial event of HR repair, the mechanism regulating the recruitment of CtIP to DSB sites remains largely unknown. Here, we show that acidic nucleoplasmic DNA‐binding protein 1 (And‐1) forms complexes with CtIP as well as other repair proteins, and is essential for HR repair by regulating DSB end resection. Furthermore, And-1 is recruited to DNA DSB sites in a manner dependent on MDC1, BRCA1 and ATM, down-regulation of And-1 impairs end resection by reducing the …

Frontal Plane Comparison Between Drop Jump And Vertical Jump: Implications For The Assessment Of Acl Risk Of Injury, Guilherme Manna Cesar, Curtis L. Tomasevicz, Judith M. Burnfield

Athletic Performance Research

The potential to use the vertical jump (VJ) to assess both athletic performance and risk of anterior cruciate ligament (ACL) injury could have widespread clinical implications since VJ is broadly used in high school, university, and professional sport settings. Although drop jump (DJ) and VJ observationally exhibit similar lower extremity mechanics, the extent to which VJ can also be used as screening tool for ACL injury risk has not been assessed. This study evaluated whether individuals exhibit similar knee joint frontal plane kinematic and kinetic patterns when performing VJs compared with DJs. Twenty-eight female collegiate athletes performed DJs and VJs. …

Weight Control Practices Of Division I National Collegiate Athletic Association Athletes, Patrick B. Wilson, Leilani Madrigal, Judith M. Burnfield

Athletic Performance Research

Objectives: Altering body weight can have substantial effects on an athlete’s performance and well-being. Limited information is available describing the weight control practices of Division I National Collegiate Athletic Association (NCAA) athletes.

Methods: Weight control practices data from 188 (138 male and 50 female; 18-23 y) Division I NCAA athletes were analyzed as a part of this cross-sectional, retrospective study. Participants completed questionnaires on weight control practices and weight control nutrition knowledge at the end of their season and were classified into weight-sensitive and less weight-sensitive sports.

Results: A higher proportion of females attempted to lose weight than males among …

Nitrogenase Femoco Investigated By Spatially Resolved Anomalous Dispersion Refinement, Thomas Spatzal, Julia Schlesier, Eva-Maria Burger, Daniel Sippel, Limei Zhang, Susana L.A. Andrade, Douglas C. Rees, Oliver Einsle

Department of Biochemistry: Faculty Publications

The [Mo:7Fe:9S:C] iron-molybdenum cofactor (FeMoco) of nitrogenase is the largest known metal cluster and catalyses the 6-electron reduction of dinitrogen to ammonium in biological nitrogen fixation. Only recently its atomic structure was clarified, while its reactivity and electronic structure remain under debate. Here we show that for its resting S 1⁄4 3/2 state the common iron oxidation state assignments must be reconsidered. By a spatially resolved refinement of the anomalous scattering contributions of the 7 Fe atoms of FeMoco, we conclude that three irons (Fe1/3/7) are more reduced than the other four (Fe2/4/5/6). Our data are in agreement with the …

A Critical Role Of Cxcr2 Pdz Motif-Mediated Interactions In Endothelial Progenitor Cell Homing And Angiogenesis, Yuning Hou

Wayne State University Dissertations

Bone marrow-derived endothelial progenitor cells (EPCs) participate in postnatal vascularization in response to growth factors, cytokines, and chemokines. Chemokine receptor CXCR2 and its cognate ligands are reported to mediate EPC recruitment and angiogenesis. CXCR2 possesses a consensus PSD-95/DlgA/ZO-1 (PDZ) motif at its carboxyl terminus. The PDZ motif has been reported to regulate cellular signaling and functions. Here we investigated the potential role of the PDZ motif in CXCR2-mediated EPC motility and angiogenesis. We have found that introducing exogenous CXCR2 C-terminus significantly attenuated in vitro EPC migration and angiogenic activities in response to CXCR2 ligands, as well as in vivo EPC …

The Development Of Peptide Ligands To Target H69 Rrna, Danielle Nicole Dremann

Wayne State University Dissertations

ABSTRACT

THE DEVELOPMENT OF PEPTIDE LIGANDS TO TARGET H69

by

DANIELLE NICOLE DREMANN

December 2015

Advisor: Prof. Christine S. Chow

Major: Chemistry (Biochemistry)

Degree: Doctor of Philosophy

In the development of peptide ligands to target H69, SPPS and ESI MS was used to determine if 1) peptides could bind to modified H69 and 2) if increased affinity for the target RNA could be enhanced with modification. An alanine and arginine scan was synthesized and tested for this determination. Selected peptides were then tested using biophysical techniques such as circular dichroism and isothermal titration calorimetry. An assay was also designed to …

The Effects Of Courtship And Pairing Behavior On The Nonapeptide And Noradrenergic Systems Of Adult Male And Female Zebra Finches, Erin Lowrey Ondercin

Wayne State University Dissertations

Social relationships are complex and likely involve the multiple neural circuits, including those involved in learning, memory, motivation, and attention. Two neurotransmitter pathways highly involved in these neural circuits are norepinephrine (NE) and the nonapeptides, vasopressin (AVP) and oxytocin (OT). There is extensive research implicating a role for the nonapeptides in trust, sociality, parental care, and romantic relationships. There is little direct evidence for the role of nonapeptides in monogamous relationships in any species other than the prairie vole (Goodson 2013). However, there is evidence that nonapeptides are important in pair bonding for both male and female zebra finches (Lowrey …

Bacterial Forensics: Revolutionizing Biochemical Analysis, Caitlin Cain

AUCTUS: The Journal of Undergraduate Research and Creative Scholarship

Eva Childrey is a junior forensic science and chemistry double major working in Dr. Eh- rhardt’s research laboratory at VCU. The main goal of the research conducted in this laboratory is to explore the lipid profiles of different bacterial species.

Students' Perception Of Educational Environment At Aga Khan University Medical College, Karachi, Pakistan, Rehana Rehman, Kulsoom Ghias, Syeda Sadia Fatima, Mehwish Hussain, Faiza Alam

Department of Biological & Biomedical Sciences

Objective: To assess educational environment in Aga Khan University Medical College (AKUMC) by Dundee Ready Educational Environmental Measure (DREEM) scale.
Methods: A cross-sectional survey of students at the AKUMC with simple random sampling was carried out from June 2014 till March 2015. Responses in five subscales were used to calculate DREEM scores.
Results: The average DREEM score was 125.77±16.8 with a reliability of 91.3%. With regards to subscales, on the 12-item students' perceptions of learning (PoL) subscale, the maximum score was 48; 11 items of students' perceptions of teachers (PoT) had a maximum score of 44; students' academic self-perceptions (ASP) …

Regulation Of Cytochrome C Functions By Phosphorylation, Gargi Mahapatra

Wayne State University Dissertations

The long term goal of my thesis research is to understand how tissue-specific

phosphorylations on the small mitochondrial protein, cytochrome c (Cytc), regulate its

functions, under both physiologically healthy and stressed conditions, and to identify the

cell signaling pathways targeting Cytc. Cytc is a functionally diverse protein that carries

electrons in the electron transport chain and plays a critical role in cellular apoptosis, two

diverse pathways that maintain cellular health that are active under diverse conditions.

Since Cytc plays a pivotal role in both these highly divergent pathways, regulation of the

protein is very important—phosphorylation of the protein under physiological …

Investigating The Role Of Dna Polymerase Beta In The Aging Phenotype Of Down Syndrome, Aqila Ahmed Ahmed

Wayne State University Dissertations

Down syndrome (DS) is a chromosomal condition characterized by accelerated aging that has yet to be directly linked to a DNA repair defect. Reduced PolB and unrepaired damage from oxidative stress observed in DS, point toward defective base excision repair (BER). In this study, we report that low PolB transcript correlates with increased markers of senescence. The gene dosage effect of Trisomy 21 is likely the source for PolB downregulation. We show that the HSA21-localized miR-155 overexpression correlates with a decrease in Creb1 and PolB, thus establishing a putative regulatory pathway. Data from the DS mouse model, Ts65Dn, reveal low …

Studies Towards Broadening The Substrate Profile And Regulation Of Histone Deacetylase 1, Dhanusha Ashanthi Nalawansha

Wayne State University Dissertations

Aberrant expression of histone deacetylase 1 (HDAC1) is implicated in multiple diseases, including cancer. As a consequence, HDAC1 has emerged as an important therapeutic target for drug development. HDAC1 regulates key cellular processes, such as cell proliferation, apoptosis, and cell survival, by deacetylating both histone and non-histone substrates. Due to the lack of simple tools to identify physiological substrates of HDAC1, the full spectrum of HDAC1 activities in the cell remains unclear. Here, we employed a substrate trapping strategy to identify cellular substrates of HDAC1. Using this approach, we identified mitosis-related protein Eg5 as a substrate. HDAC1 colocalizes with Eg5 …

The Unusual Substrate Specificity Of A Virulence Associated Serine Hydrolase From The Highly Toxic Bacterium, Francisella Tularensis, Alexander M. Farberg, Whitney K. Hart, R. Jeremy Johnson

Scholarship and Professional Work - LAS

Francisella tularensis is the causative agent of the highly, infectious disease, tularemia. Amongst the genes identified as essential to the virulence of F. tularensis was the proposed serine hydrolase FTT0941c. Herein, we purified FTT0941c to homogeneity and then characterized the folded stability, enzymatic activity, and substrate specificity of FTT0941c. Based on phylogenetic analysis, FTT0941c was classified within a divergent Francisella subbranch of the bacterial hormone sensitive lipase (HSL) superfamily, but with the conserved sequence motifs of a bacterial serine hydrolase. FTT0941c showed broad hydrolase activity against diverse libraries of ester substrates, including significant hydrolytic activity across alkyl ester substrates from …

Thermodynamics And Kinetics Of The Three-Way Junction Of Phi29 Motor Prna And Its Assembly Into Nanoparticles For Therapeutic Delivery To Prostate Cancer, Daniel W. Binzel

Theses and Dissertations--Pharmacy

The emerging field of RNA nanotechnology necessitates creation of functional RNA nanoparticles, but has been limited by particle instability. Previously, it was found the three-way junction (3WJ) of the Phi29 DNA packaging motor pRNA was found to be ultra-stable and assemble in solution without the presence of metal ions. The three-way junction is composed of three short oligo RNA strands and proven to be thermodynamically stable. Here the assembly mechanism, thermodynamic and enzymatic stabilities, and kinetics are examined in order to understand the stability behind this unique motif. Thermodynamic and kinetics studies found that the pRNA 3WJ formed out of …

Investigating And Expanding The Functionality Of Rna Catalysts: Studies Of The Hepatitis Delta Virus, The Hammerhead, And The Aminoacyl-Trna Synthetase-Like Ribozymes, Ji Chen

Open Access Dissertations

Ribozymes, just like protein enzymes, catalyze diverse chemical reactions. The first goal of this dissertation is to understand the mechanism of ribozyme-mediated phosphodiester cleavage reaction. Biochemical assays and X-ray crystallography were used for probing the active site of two small self-cleaving ribozymes, the hepatitis delta virus (HDV) ribozyme and the hammerhead ribozyme. Results presented here suggest that divalent metal ions play critical roles in the catalytic mechanisms of both the HDV and the hammerhead ribozymes. In the HDV ribozyme, the result is consistent with an active site Mg2+ being directly involved in catalysis. In the hammerhead ribozyme, however, Mg2+ ions …

An Investigation Of Juvenile Alewife (Alosa Pseudoharengus) Habitat Use And Growth Using Natural Markers, Gregory Norman Labonte Ms

All Student Scholarship

This research attempts to connect patterns in growth and migration of an anadromous species. The goal of this research was to understand habitat movements and growth of juvenile alewives (Alosa pseudoharengus) in the Penobscot Estuary and Bay through the use of otolith microchemistry, otolith growth increments, and a laboratory stable isotope turnover study. Understanding the connection between growth and movement of juvenile alewives may lead to more accurate and sophisticated conservation and restoration methods for anadromous species.

Transcriptomic Analysis Of Staphylococcal Srnas: Insights Into Species-Specific Adaption And The Evolution Of Pathogenesis, William H. Broach, Andy Weiss, Lindsey N. Shaw

Molecular Biosciences Faculty Publications

Next-generation sequencing technologies have dramatically increased the rate at which new genomes are sequenced. Accordingly, automated annotation programs have become adept at identifying and annotating protein coding regions, as well as common and conserved RNAs. Additionally, RNAseq techniques have advanced our ability to identify and annotate regulatory RNAs (sRNAs), which remain significantly understudied. Recently, our group catalogued and annotated all previously known and newly identified sRNAs in several Staphylococcus aureus strains. These complete annotation files now serve as tools to compare the sRNA content of S. aureus with other bacterial strains to investigate the conservation of their sRNomes. Accordingly, in …

Potential Role Of Pctaire-2, Pctaire-3 And P-Histone H4 In Amyloid Precursor Protein-Dependent Alzheimer Pathology, Dale Chaput, Lisa Kirouac, Stanley M. Stevens Jr., Jaya Padmanabhan

Molecular Biosciences Faculty Publications

Amyloid Precursor Protein (APP) is regulated in a mitosis-specific manner and plays a role in proliferative signaling in cells. Though APP-derived Aβ generation has a well-established role in neurodegeneration, the mechanistic role of APP in this process is not fully understood. Here, we performed an unbiased, comprehensive analysis of the phosphoproteome signature in APP-null neuroblastoma cells (B103) compared to those expressing APP-695 isoform (B103-695) to determine if APP expression affects protein phosphorylation. Stable isotope labeling by amino acids in cell culture (SILAC) followed by mass spectrometry-based phosphoproteomic analysis with PolyMAC identified a total of 2,478 phosphopeptides in the B103 and …

Antibodies Against A Secreted Product Of Staphylococcus Aureus Trigger Phagocytic Killing, Lena Thomer, Carla Emolo, Vilasack Thammavongsa, Hwan Keun Kim, Molly E. Mcadow, Wenqi Yu, Matthew Kieffer, Olaf Schneewind, Dominique Missiakas

Molecular Biosciences Faculty Publications

Host immunity against bacteria typically involves antibodies that recognize the microbial surface and promote phagocytic killing. Methicillin-resistant Staphylococcus aureus (MRSA) is a frequent cause of lethal bloodstream infection; however, vaccines and antibody therapeutics targeting staphylococcal surface molecules have thus far failed to achieve clinical efficacy. S. aureus secretes coagulase (Coa), which activates host prothrombin and generates fibrin fibrils that protect the pathogen against phagocytosis by immune cells. Because of negative selection, the coding sequence for the prothrombin-binding D1-D2 domain is highly variable and does not elicit cross-protective immune responses. The R domain, tandem repeats of a 27-residue peptide that bind …