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A Multilaboratory Comparison Of Calibration Accuracy And The Performance Of External References In Analytical Ultracentrifugation, Huaying Zhao, Rodolfo Ghirlando, Carlos Alfonso, Fumio Arisaka, Ilan Attali, David L. Bain, Marina M. Bakhtina, Donald F. Becker, Gregory J. Bedwell, Ahmet Bekdemir, Tabot M. D. Besong, Catherine Birck, Chad A. Brautigam, William Brennerman, Olwyn Byron, Agnieszka Bzowska, Jonathan B. Chaires, Catherine T. Chaton, Helmut Cölfen, Keith D. Connaghan, Kimberly A. Crowley, Ute Curth, Tina Daviter, William L. Dean, Ana I. Díez, Christine Ebel, Debra M. Eckert, Leslie E. Eisele, Edward Eisenstein, Patrick England, Carlos Escalante, Jeffrey A. Fagan, Robert Fairman, Ron M. Finn, Wolfgang Fischle, José García De La Torre, Jayesh Gor, Henning Gustafsson, Damien Hall, Stephen E. Harding, José G. Hernández Cifre, Andrew B. Herr, Elizabeth E. Howell, Richard S. Isaac, Shu-Chuan Jao, Davis Jose, Soon-Jong Kim, Bashkim Kokona, Jack A. Kornblatt, Dalibor Kosek, Elena Krayukhina, Daniel Krzizike, Eric A. Kusznir, Hyewon Kwon, Adam Larson, Thomas M. Laue, Aline Le Roy, Andrew P. Leech, Hauke Lilie, Karolin Lugar, Juan R. Luque-Ortega, Jia Ma, Carrie A. May, Ernest L. Maynard, Anna Modrak-Wojcik, Yee-Foong Mok, Norbert Mücke, Luitgard Nagel-Steger, Geeta J. Narlikar, Masanori Noda, Amanda Nourse, Tomas Obsil, Chad K. Park, Jin-Ku Park, Peter D. Pawelek, Erby E. Perdue, Stephen J. Perkins, Matthew A. Perugini, Craig L. Peterson, Martin G. Peverelli, Grzegorz Piszczek, Gali Prag, Peter E. Prevelige, Bertrand D. E. Raynal, Lenka Rezabkova, Klaus Richter, Alison E. Ringel, Rose Rosenberg, Arthur J. Rowe, Arne C. Rufer, David J. Scott, Javier G. Seravalli, Alexandra S. Solovyova, Renjie Song, David Staunton, Catlin Stoddard, Katherine Stott, Holger M. Strauss, Werner W. Streicher, John P. Sumida, Sarah G. Swygert, Roman H. Szczepanowski, Ingrid Tessmer, Ronald T. Toth Iv., Ashutosh Tripathy, Susumu Uchiyama, Stephen F. W. Uebel, Satoru Unzai, Anna Vitlin Gruber, Peter H. Von Hippel, Christine Wandrey, Szu-Huan Wang, Steven E. Weitzel, Beata Wielgus-Kutrowska, Cynthia Wolberger, Martin Wolff, Edward Wright, Yu-Sung Wu, Jacinta M. Wubben, Peter Schuck

Physiology and Biophysics Publications and Presentations

Analytical ultracentrifugation (AUC) is a first principles based method to determine absolute sedimentation coefficients and buoyant molar masses of macromolecules and their complexes, reporting on their size and shape in free solution. The purpose of this multi-laboratory study was to establish the precision and accuracy of basic data dimensions in AUC and validate previously proposed calibration techniques. Three kits of AUC cell assemblies containing radial and temperature calibration tools and a bovine serum albumin (BSA) reference sample were shared among 67 laboratories, generating 129 comprehensive data sets. These allowed for an assessment of many parameters of instrument performance, including accuracy …

The Effect Of Optogenetic Manipulation Of Ss Interneurons Within Malformed, Epileptogenic Cortex, Nicole Ekanem

Theses and Dissertations

A large percentage of individuals with intractable epilepsies have an accompanying cortical malformation, the underlying cellular mechanisms of which are poorly understood. It is known however that in an animal model for one such malformation, polymicrogyria, epileptogenesis occurs most easily from an adjacent area termed the paramicrogyral region (PMR). Previous studies implicate SS interneurons as a potential contributor to this pathology, which lead to our hypothesis: in PMR, SS interneurons exert a higher modulatory influence on excitatory pyramidal cells, as compared to the same by SS interneurons within homologous control cortex.

Using a freeze-lesion model for polymicrogyria in transgenic mice …

Characterization Of A Novel Protease In Staphylococcus Aureus, Adam L. Johnson

Theses and Dissertations

A newly discovered cysteine protease, Prp, has been shown to perform an essential, site-specific cleavage of ribosomal protein L27 in Staphylococcus aureus. In Firmicutes and related bacteria, ribosomal protein L27 is encoded with a conserved N-terminal extension that must be removed to expose residues critical for ribosome function. Uncleavable and pre-cleaved variants were unable to complement an L27 deletion in S. aureus, indicating that this N-terminal processing event is essential and likely plays an important regulatory role. The gene encoding the responsible protease (prp) has been shown to be essential, and is found in all organisms …

High-Throughput Data Analysis: Application To Micronuclei Frequency And T-Cell Receptor Sequencing, Mateusz Makowski

Theses and Dissertations

The advent of high-throughput sequencing has brought about the creation of an unprecedented amount of research data. Analytical methodology has not been able to keep pace with the plethora of data being produced. Two assays, ImmunoSEQ and the cytokinesisblock micronucleus (CBMN), that both produce count data and have few methods available to analyze them are considered.

ImmunoSEQ is a sequencing assay that measures the beta T-cell receptor (TCR) repertoire. The ImmunoSEQ assay was used to describe the TCR repertoires of patients that have undergone hematopoietic stem cell transplantation (HSCT). Several different methods for spectratype analysis were extended to the TCR …

Virginia Dental Journal (Vol. 92, No. 3 [I.E. 2], 2015)

Virginia Dental Journal

No abstract provided.

The Effect Of Statins On Il-33 Mediated Mast Cell Function, Marcela Taruselli

Theses and Dissertations

This study demonstrates original findings of statin effects on IL-33 stimulated mast cells. Statins are a class of drugs used to lower cholesterol production by targeting HMG CoA reductase. These commonly prescribed drugs have been shown to be immunomodulatory. In this study, we have found that pretreatment with statins has a variety of effects on IL-33 stimulated mast cells. Atorvastatin suppresses TNF and IL-6 production, while fluvastatin significantly enhances release of these proinflammatory cytokines in BMMCs. Although they have differing effects on cytokine production, both statins lowered ST2 expression on the cell surface, decreased cell viability, and enhanced expression of …

Immunotherapy Of Cancer: Reprogramming Tumor/Immune Cellular Crosstalk To Improve Anti-Tumor Efficacy, Kyle K. Payne

Theses and Dissertations

Immunotherapy of cancer has been shown to be promising in prolonging patient survival. However, complete elimination of cancer and life-long relapse-free survival remain to be major challenge for anti-cancer therapeutics. We have previously reported that ex vivo reprogramming of tumor-sensitized immune cells by bryostatin 1/ionomycin (B/I) and the gamma-chain (γ-c) cytokines IL-2, IL-7, and IL-15 resulted in the generation of memory T cells as well as CD25+ NKT cells and CD25+ NK cells. Adoptive cellular therapy (ACT) utilizing these reprogrammed immune cells protected FVBN202 mice from tumor challenge, and overcame the suppressive functions of myeloid-derived suppressor cells (MDSCs). We then …

Mast Cells In Kainate Receptor Knockout Mice, Andrea J. Elkovich

Theses and Dissertations

Kainate receptor knockout mice have unique differences within their immune system. They exhibit an attenuated T_H2 branch, while maintaining a robust T_H1 response. Specifically, blocking the formation of functional kainate receptors affects mast cells and their related pathologies. While they seem to develop and activate normally in vivo and in vitro, KAR KO mast cells release more inflammatory mediators upon degranulation. These mice experience severe anaphylactic shock due to two compounding abnormalities. First, KAR KO mast cells release significantly more histamine in vivo upon IgE-mediated activation. Second, the animals over-respond to exogenous histamine with drastic …

Characterization Of Transfer Of The Mobile Genomic Island Encoding Methicillin Resistance Among Staphylococci, Melissa D. Ray

Theses and Dissertations

The gene encoding methicillin resistance in Staphylococcus aureus (MRSA) is carried in the chromosome on a large genomic island called SCCmec and is always inserted at the att site within orfX. SCCmec has been designated a mobile genetic element but a mechanism by which it moves among different strains and species of staphylococci has never been demonstrated. This work shows that bacteriophage 80α is capable of transducing SCCmec into a recipient cell, after which it can integrate into the bacterial chromosome via homologous recombination. More importantly, this work characterizes a conjugative mechanism of SCCmec transfer. …

Enhancement Of The Placental Transmission Of Lopinavir Using A Transporter Targeted Prodrug Strategy, Meng Wang

Theses and Dissertations

Lopinavir (LPV) is a potent protease inhibitor specific for HIV-1. However, LPV has poor placental penetration due to substrate activity for efflux transporter by P-glycoprotein (P-gp). Since fatty acid transporters are highly expressed in the placenta during pregnancy, we designed fatty acid ester prodrug of lopinavir as substrates of fatty acid transporter in order to improve their uptake into placenta. Seven dicarboxylic acid esters of lopinavir have been made in our lab. The structures were characterized by ¹H-NMR, ¹³C-NMR, LC-MS/MS, HRMS, IR and melting points. After making the prodrugs, an LC-MS/MS method with high specificity and sensitivity, as …

Orientia Tsutsugamushi Ankyrin-Repeat Protein Family Targeting Of The Host Endoplasmic Reticulum, Lauren Viebrock

Theses and Dissertations

Abstract

ORIENTIA TSUTSUGAMUSHI ANKYRIN REPEAT-PROTEIN FAMILY TARGETING OF THE HOST ENDOPLASMIC RETICULUM

By Lauren VieBrock, B.S.

A dissertation submitted in partial fulfillment of the requirements for the degree of Doctor of Philosophy at Virginia Commonwealth University

Virginia Commonwealth University, 2015

Director: Jason A. Carlyon, Ph.D.

Professor

Microbiology and Immunology

Scrub typhus is an understudied, potentially fatal febrile illness, which poses threat to one billion people annually in the Asia-Pacific region. The host-pathogen interactions that facilitate the intracellular survival of the etiologic agent, Orientia tsutsugamushi, are not well understood. The Orientia tsutsugamushi genome encodes a large number of ankyrin repeat-containing proteins …

Incorporation Of Molecular Diagnostics Into Medical Laboratory Science Curriculum: Clinical Facilities Expectations. An Asynchronous, Iterative, Online Delphi Study., Barbara Kraj

Theses and Dissertations

The medical laboratory science (MLS) profession is in need for published molecular diagnostics competency-based standards and curriculum. To assess their expectations of new MLS graduates, professionals performing and supervising performance of clinical molecular assays were surveyed to rate the importance of relevant cognitive and psychomotor learning objectives. A modified, asynchronous, iterative online Delphi process was utilized for assessment of consensus on the importance of the objectives. The survey was delivered through online REDCap application. Program directors of 221 MLS programs accredited by the National Accrediting Agency for Clinical Laboratory Science (NAACLS) were asked to forward the first Delphi survey to …

General Dentists' Role In Providing Care To Very Young Children:Pediatric Dentists' Perspective, Shinjni Razdan

Theses and Dissertations

To examine the pediatric dentists’ perspective on level of care to children 0-3 years old by general dentists, an electronic survey was sent out to 5185 AAPD members and 769 responded (response rate: 14.8%). Forty-six percent agreed with a general dentist providing a dental home to children 0-3 years old. Only 24% agreed that general dentists could perform complex restorative and behavior management procedures for 0-3 years old. Younger respondents, pediatric dentists who practice in rural areas or teaching pediatric dentistry, and those who perceived adequate exposure during dental school were more likely to agree on general dentists proving a …

The Effect Of Dexamethasone On Il-33-Mediated Mast Cell Activation, Oksana I. Chernushevich

Theses and Dissertations

Dexamethasone has been shown to inhibit IgE-mediated mast cell activation, and the present research investigated its role in suppressing IL-33-mediated mast cell activation. We have found that micromolar concentrations of Dexamethasone are capable of suppressing IL-33-mediated mast cell cytokine production, on several genetic backgrounds, and in not only bone marrow derived mast cells, but also peritoneal mast cells. Intracellular staining demonstrated that Dexamethasone significantly reduces expression of the IL-33 receptor, T1/ST2, in mast cells; however, the cytokine suppression is independent of T1/ST2 downregulation. At the same time, Dexamethasone pretreatment significantly reduced ERK phosphorylation, but our data suggests that inhibition occurs …

The Role Of Adam10, Adam17, And Spag6 In Humoral Immunity And Secondary Lymphoid Tissue Architecture, Lauren Folgosa Cooley

Theses and Dissertations

ADAM10, ADAM17, and SPAG6 contribute significantly to humoral immunity and secondary lymphoid tissue architecture. ADAM10 and ADAM17 are two closely related zinc-metalloproteinases. Through cleavage of their ligands CD23 and TNF, respectively, they greatly influence IgE production and secondary lymphoid tissue architecture maintenance. Th1 prone WT strains initially exhibit increased ADAM17 and TNF yet reduced ADAM10 relative to Th2 prone WT strains. In the absence of B cell ADAM10, a compensatory increase in ADAM17 and TNF cleavage is noted only in Th1 prone C57Bl/6, not Th2 prone Balb/c. B cell TNF homeostasis is important for maintaining secondary lymphoid tissue architecture. We …

A Continuous Process Towards The Synthesis Of Quinolones, Stevara N. Clinton

Theses and Dissertations

Abstract

The development of quinolones is described from the first quinolone to the latest fluoroquinolones. Quinolones have generated considerable interest since their discovery because of their antibacterial capabilities. Analogs incorporating the 4-quinolone ring system comprise a largely expanding group of synthetic compounds. The development of antibacterial resistance has created the need for an efficient synthesis of quinolones that can be easily adapted toward the assembly of quinolone based antibacterial drugs.

There are several reported approaches to the 4-quinolone ring system. Many of these methods use expensive starting materials, require the removal of high boiling solvents, or use high temperature conditions …

Development Of Novel Chimeric Epitope Based Diagnostic Antigens And Vaccines For Lyme Disease, Lee D. Oliver Jr.

Theses and Dissertations

Lyme disease (LD) is the leading arthropod-borne disease in North America with 300-600,000 cases each year. There are currently no approved human LD vaccines. Outer surface protein C (OspC) has emerged as a leading vaccine candidate and an attractive diagnostic marker due to its antigenicity and expression early in infection. Several chimeric, epitope based OspC derived proteins were generated. The constructs were found to be highly immunogenic in mice and vaccination induced complement-dependent bactericidal antibodies. These results suggest that a broadly protective polyvalent OspC epitope based vaccine can be produced. Currently, LD diagnostic approaches are unreliable and unable to differentiate …

Commonwealth Times 2014-10-27

Commonwealth Times, 1969-

No abstract provided.

Investigations Of Novel Mechanisms Of Action For Anti-Bacterial And Anti-Cancer Agent Development, Jenson Verghese

Theses and Dissertations

The development of drugs and therapeutic agents for combating infections and human malignancies continues to be a forefront area in both academic and industrial research. This is driven by the rapid emergence of multi-drug resistant bacterial strains and accumulating mutations in cancer targets that is quickly rendering our current arsenal of drugs ineffective for these therapies. Unless new drugs with novel mechanisms of action are identified and developed at a faster pace, we face a losing battle in managing these diseases. The first part of this work concerns with the natural product Simocyclinone D8 (SD8). Simocyclinone D8 is an angucyclinone …

A Retrospective Study Of Operating Room Utilization And Efficiency In A Pediatric Dental Residency Program, Brian Burke

Theses and Dissertations

Purpose: The purpose was to assess and understand operating room (OR) utilization and efficiency in a pediatric dental residency program. Methods: A retrospective study was performed using chart extraction from 778 patients completed by both pediatric dentistry faculty (n=7) and residents (n=17) in an ambulatory care setting over a 32 month period (between July 2010 and March 2013). Patterns in OR usage time were determined by documenting various timing metrics (start and stop times for anesthesia, start and stop times for the dental procedure, times for throat pack in and out), noting patient information (age and ASA patient classification status), …

Patient Discomfort In The Icu: Ett Movement Effects, Virginia Hamilton

Theses and Dissertations

Critically ill patients who require MV are at risk for a number of complications, including the development of ventilator-associated events (VAE) and agitation that may require the use of sedation. Patients experience anxiety and discomfort during mechanical ventilation from a variety of sources including unfamiliar breathing assistance and an inability to communicate anxiety and pain verbally, but a primary cause of discomfort identified by these patients is the simply the presence of the endotracheal tube (ETT). Discomfort often leads to agitation and may be exacerbated by ETT movement. Management of agitation typically involves the use of sedative therapy and has …

Evaluation Of Maxillary Molar Furcations, Clinical Measurements Versus Cone Beam Computed Tomography, Jessica Allen

Theses and Dissertations

BACKGROUND: The use of three-dimensional imaging has shown to provide advantages to the clinician in assessing bone morphology. The aim of this study will be to compare the diagnostic efficacy of cone beam computed tomography (CBCT) versus diagnostic clinical measurements in patients presenting with furcation involved maxillary first molars. METHODS: The study population included 20 patients with 34 maxillary first molar teeth with furcation involvement. Clinical horizontal and vertical probing measurements were compared to CBCT measurements taken by two calibrated examiners. RESULTS: Horizontal measurements showed a significant difference between Glickman class II and class III. There were no statistical significant …

An Examination Of The Relationship Of Abo Blood Group And Lifespan In A Hospitalized Population In The Southeastern United States, Tara Moon

Theses and Dissertations

The clinical significance of ABO blood group is evident and universally accepted with regards to blood transfusion and pregnancy; however, the importance of ABO blood group as it relates to other diseases or disorders and overall mortality is not fully understood by the scientific community. Many studies have suggested associations between blood groups and disease, but consensus has not been reached regarding overall survival or longevity. This epidemiological, retrospective review of ABO blood group and age at the time of death in a hospitalized population in the Southeastern United States is the first multi-site study to examine this relationship. The …

Utility Of Surveillance Blood Cultures In Patients Undergoing Hematopoietic Stem Cell Transplantation, Sameeh S. Ghazal, Michael P. Stevens, Gonzalo M. L. Bearman, Michael B. Edmond

Internal Medicine Publications

Background Surveillance blood cultures are often obtained in hematopoietic stem cell transplant (HSCT) patients for detection of bloodstream infection. The major aims of this retrospective cohort study were to determine the utility of the practice of obtaining surveillance blood cultures from asymptomatic patients during the first 100 post-transplant days and to determine if obtaining more than one positive blood culture helps in the diagnosis of bloodstream infection.

Methods We conducted a 17-month retrospective analysis of all blood cultures obtained for patients admitted to the hospital for HSCT from January 2010 to June 2011. Each patient’s clinical course, vital signs, diagnostic …

Inhibition Of Methicillin-Resistant Staphylococcus Aureus-Induced Cytokines Mrna Production In Human Bone Marrow Derived Mesenchymal Stem Cells By 1,25-Dihydroxyvitamin D3, Aparna Maiti, William A. Jiranek

Orthopaedic Surgery Publications

Background Methicillin-resistant Staphylococcus aureus (MRSA) is the predominant cause of bone infection. Toll like receptors (TLRs) are an important segments of host response to infection and are expressed by a variety of cells including human mesenchymal stem cells (hMSCs). The active form of Vitamin D, 1,25-dihydroxyvitamin D3 (1,25(OH)2D3) has potent immunoregulatory properties, but the mechanism remains poorly understood. The genomic action of 1,25(OH)2D3 is mediated by vitamin D receptor (VDR), hormone-regulated transcription factor. VDR interacts with co-activators and co-repressors are associated with chromatin histone modifications and transcriptional regulation. The aim of our study is to explore MRSA-induced TLRs-mediated pro-inflammatory cytokines …

Medical Literary Messenger (Vol. 2, No. 1, Fall 2014)

Medical Literary Messenger

Quiet Please Sickness / Cathleen Calbert -- Helping a seven year old with inoperable brain cancer write a poem for her sister / Kate Peterson -- Asking My Liver for Forgiveness / Rob Cook -- Synesthesia 2 / Catherine Harnett -- Tic / Molly Lazer -- The Bridge / Alexander Schloe -- Mother’s Milk and Cystic Fibrosis / Lizabeth Berkeley -- Fanny Brawne / William Miller -- artwork / Venus -- Cure / Anne Spollen -- Gene / Kate Peterson -- The Love Song of Elliot Blue / Jacqueline Kirkpatrick -- “Next” / Michael P. Stevens -- Structures / C.I.L. …

Sperm-Associated Antigen 6 (Spag6) Deficiency And Defects In Ciliogenesis And Cilia Function: Polarity, Density, And Beat, Maria E. Teves, Patrick R. Sears, Wei Li, Zhengang Zhang, Waixing Tang, Lauren Van Reesema, Richard M. Costanzo, C. William Davis, Michael R. Knowles, Jerome F. Strauss Iii, Zhibing Zhang

Obstetrics and Gynecology Publications

SPAG6, an axoneme central apparatus protein, is essential for function of ependymal cell cilia and sperm flagella. A significant number of Spag6-deficient mice die with hydrocephalus, and surviving males are sterile because of sperm motility defects. In further exploring the ciliary dysfunction in Spag6-null mice, we discovered that cilia beat frequency was significantly reduced in tracheal epithelial cells, and that the beat was not synchronized. There was also a significant reduction in cilia density in both brain ependymal and trachea epithelial cells, and cilia arrays were disorganized. The orientation of basal feet, which determines the direction of axoneme …

Impact Of Appropriate Antimicrobial Therapy For Patients With Severe Sepsis And Septic Shock – A Quality Improvement Study, Paula K. O. Yokota, Alexandre R. Marra, Marines D. V. Martino, Elivane S. Victor, Marcelino S. Durão, Michael B. Edmond, Oscar F. P. Dos Santos

Internal Medicine Publications

Background

There is ample literature available on the association between both time to antibiotics and appropriateness of antibiotics and clinical outcomes from sepsis. In fact, the current state of debate surrounds the balance to be struck between prompt empirical therapy and care in the choice of appropriate antibiotics (both in terms of the susceptibility of infecting organism and minimizing resistance arising from use of broad-spectrum agents). The objective of this study is to determine sepsis bundle compliance and the appropriateness of antimicrobial therapy in patients with severe sepsis and septic shock and its impact on outcomes.

Material

This study was …

Impact Of Appropriate Antimicrobial Therapy For Patients With Severe Sepsis And Septic Shock – A Quality Improvement Study, Paula K. O. Yokota, Alexandre R. Marra, Marines D. V. Martino, Elivane S. Victor, Marcelino S. Durão, Michael B. Edmond, Oscar F. P. Dos Santos

Internal Medicine Publications

Background

There is ample literature available on the association between both time to antibiotics and appropriateness of antibiotics and clinical outcomes from sepsis. In fact, the current state of debate surrounds the balance to be struck between prompt empirical therapy and care in the choice of appropriate antibiotics (both in terms of the susceptibility of infecting organism and minimizing resistance arising from use of broad-spectrum agents). The objective of this study is to determine sepsis bundle compliance and the appropriateness of antimicrobial therapy in patients with severe sepsis and septic shock and its impact on outcomes.

Material

This study was …

Clinical And Microbiologic Efficacy Of A Water Filter Program In A Rural Honduran Community, Jaclyn Arquiette, Michael P. Stevens, Jean M. Rabb, Kakotan Sanogo, Patrick Mason, Gonzalo Bearman

VCU Health Publications

Water purification in the rural Honduras is a focus of the nonprofit organization Honduras Outreach Medical Brigade Relief Effort (HOMBRE). We assessed water filter use and tested filter microbiologic and clinical efficacy. A 22-item questionnaire assessed water sources, obtainment/storage, purification, and incidence of gastrointestinal disease. Samples from home clay-based filters in La Hicaca were obtained and paired with surveys from the same home. We counted bacterial colonies of four bacterial classifications from each sample. Sixty-five surveys were completed. Forty-five (69%) individuals used a filter. Fifteen respondents reported diarrhea in their home in the last 30 days; this incidence was higher …