Cell Biology Commons^™

Prospective Study Of The Feasibility Of Point-Of-Care Testing Strategy For Carbapenem-Resistant Organism Detection, Rahul Pannala, Bruce Baldwin, Vijay Aluru, Thomas E. Grys, Jordan Holmes, Laurence J. Miller, M. Edwyn Harrison, Cuong C. Nguyen, Fred C. Tenover, David Persing, Douglas O. Faigel

Biology Faculty Publications

Background/aims: In an investigator-initiated, prospective study, we evaluated the feasibility of a five-gene sequence point-of-care (POC) testing strategy (Xpert CARBA-R Assay, Cepheid Inc., Sunnyvale, CA, USA), compared to reference laboratory PCR (48 – 72 hours turnaround time, two gene sequences), in patients undergoing endoscopic retrograde cholangiopancreatography (ERCP) and in a hospital outbreak investigation.

Methods: After informed consent, patients undergoing ERCP (September 2015 – April 2016, n = 191) at Mayo Clinic and potential hospital contacts (n = 9) of an index carbapenem-resistant organism (CRO)-positive inpatient were included. Two rectal swabs, one each for reference and POC assays were obtained. The …

Range Expansion And The Origin Of Usa300 North American Epidemic Methicillin-Resistant Staphylococcus Aureus, Lavanya Challagundla, Isabella A. Tickler, Xavier Didelot, David C. Coleman, Anna C. Shore, Geoffrey W. Coombs, Daniel O. Sordelli, Eric L. Brown, Robert Skov, Anders Rhod Larsen, Jinnethe Reyes, Iraida E. Robledo, Guillermo J. Vazquez, Paul Rivera, Paul D. Fey, Kurt Stevenson, Shu-Hua Wang, Barry N. Kreiswirth, Jose R. Mediavilla, Cesar A. Arias, Paul J. Planet, Rathel L. Nolan, Fred C. Tenover, Richard V. Goering, D. Ashley Robinson

Biology Faculty Publications

The USA300 North American epidemic (USA300-NAE) clone of methicillin-resistant Staphylococcus aureus has caused a wave of severe skin and soft tissue infections in the United States since it emerged in the early 2000s, but its geographic origin is obscure. Here we use the population genomic signatures expected from the serial founder effects of a geographic range expansion to infer the origin of USA300-NAE and identify polymorphisms associated with its spread. Genome sequences from 357 isolates from 22 U.S. states and territories and seven other countries are compared. We observe two significant signatures of range expansion, including decreases in genetic diversity …

The Effects Of Increased Camp Levels On Neuronal Differentiation In Murine Embryonic Stem Cells, And The Creation Of A Crispr-Induced C.1252c>T Point Mutation In The Adcy5 Gene, Elizabeth Zepeda

Cal Poly Humboldt theses and projects

ADCY5-related dyskinesia is a rare movement disorder with early onset in childhood and adolescence. Previous studies linked this disease to various point mutations in the ADCY5 gene. Recent studies show that two of the point mutations cause an increase in cyclic adenosine monophosphate (cAMP) levels. However, it remains unknown how increased levels of cAMP result in the phenotypes associated with this disease. My study examines the effects of increased cAMP levels on neuronal differentiation of mouse embryonic stem cells (mESCs). My experiments demonstrated successful differentiation of mESCs into the dopaminergic neuronal lineage, indicated by the presence of Tuj 1 (a …

The Mechanism Responsible For Mechanically Enhanced Cancer Cell Invasion, Alexander Nicholas Gasparski

Wayne State University Dissertations

Cell invasion is an important process utilized by cancer cells to progress through the metastatic cascade to form deadly secondary tumors. This process can be influenced by the wide array of biomechanical forces that cancer cells within and around a tumor face in their microenvironment. It is not completely clear how these forces, either alone or simultaneously combined with other forces, can impact the metastatic capacity of cancer cells. To address this, we have utilized an in vitro mechano-invasion assay to mimic a transient tugging force that exists within the tumor microenvironment caused by the remodeling of the extracellular matrix …

Identifying The Role Of The Type-Ii Transmembrane Serine Protease Tmprss13 In Breast Cancer, Andrew Stevan Murray

Wayne State University Dissertations

Breast cancer is the most frequently diagnosed cancer and the second leading cause of cancer death in women in the United States. Breast cancer progression is accompanied by increased expression of extracellular and cell surface proteases that are capable of degrading the extracellular matrix as well as cleaving and activating downstream targets. These proteolytic processes are critically involved in modifying the tissue microenvironment of the breast, which is necessary for cancer cell invasion and eventual dissemination of cancer cells to other organs. Therefore, identifying novel proteases that promote tumor progression is critical to create new approaches for developing improved breast …

Controlling Platelet Secretion To Modulate Hemostasis And Thrombosis, Smita Joshi

Theses and Dissertations--Molecular and Cellular Biochemistry

Upon vascular injury, activated blood platelets fuse their granules to the plasma membrane and release cargo to regulate the vascular microenvironment, a dynamic process central to platelet function in many critical processes including hemostasis, thrombosis, immunity, wound healing, angiogenesis etc. This granule- plasma membrane fusion is mediated by a family of membrane proteins- Soluble N-ethyl maleimide Attachment Receptor Proteins(SNAREs). SNAREs that reside on vesicle (v-SNAREs) /Vesicle-Associated Membrane Proteins(VAMPs) interact with target/t-SNAREs forming a trans-bilayer complex that facilitates granule fusion. Though many components of exocytic machinery are identified, it is still not clear how it could be manipulated to prevent …

Elucidating The Role Of Nidogen In The Fusion Of The Choroid Fissure, Nicholas W. Carrara

Theses and Dissertations--Biology

In the developing embryo, the timely fusion of opposing epithelial sheets into one uniform layer denotes the completion of several developmental events. Failure of this epithelial sheet fusion event (ESF) within the choroid fissure (CF) is associated with the congenital disorder Ocular Coloboma, and is one of the leading causes of pediatric blindness. A requirement for a highly coordinated dismantling of the basement membrane (BM) to allow for fusion to occur is undoubted, however the underlying mechanisms of this process are poorly understood. Due to its BM crosslinking capabilities, I have hypothesized that the regulation of nidogen plays a crucial …

Wnt Secretion Proteins Modulate Rankl-Induced Expression Of Aire In Thymic Epithelial Cells, Daniel Pollack

Dissertations and Theses

Thymic epithelial cells (TEC) are essential for a proper adaptive immune response by regulating thymocyte development and establishing central tolerance. In the thymus, TECs differentially express Wnt proteins, which activate canonical and non-canonical Wnt signaling pathways. Wnt signaling is thought to regulate cell survival, proliferation, and development although the direct molecular mechanisms in TECs have yet to be elucidated. The inducible inhibition of canonical Wnt signaling with Dkk1 leads to a rapid loss of TEC progenitors as well as a decline in mature Aire-expressing mTECs. Therefore, we explore the role of Wnt ligands potentially responsible for stimulating and/or regulating Wnt …

The Effect Of Stress Induced Premature Senescence On The Expression Of Heterogeneous Ribonucleoieoprotein, Yuriy Pechenyy

Dissertations and Theses

The role of heterogeneous nuclear ribonucleoproteins (hnRNP) in cellular senescence is yet to be defined. Cellular senescence is a terminal growth arrest in somatic cells. It is thought to be the consequence of telomeric shortening that acts as a DNA damage signal. Conversely, cells induced into premature senescence (SIPS) by oxidative stress, is independent of telomere attrition. Premature senescence has been proposed to be physiologically relevant as it can be induced by treatment with chemotherapeutic agents. In particular, we are studying the roles of hnRNP A1 and A2 in the maintenance of the senescence phenotype. hnRNPs are a family of …

Bone Marrow Derived Progenitor Cells And Their Contributions To The Thymic Stroma, Mohammed Hoque

Dissertations and Theses

The thymus serves as the primary lymphoid organ responsible for the development and selection of a self-tolerant T cell repertoire. In paradox to its critical functions for the adaptive immune response, the thymus undergoes a profound age associated decline beginning in early adult life resulting in significant decline in T-cell function.Thymic epithelial cells (TECs) are the most critical component of the thymic microenvironment and undergo rapid turn-over, so understanding the cellular mechanisms responsible for the maintenance of TEC number and organization will be critical in counteracting age associated involution, particularly in cancer patients, due to enhanced degeneration in response to …

An Optimized Solid-Phase Reduction And Capture Strategy For The Study Of Reversibly-Oxidized Cysteines And Its Application To Metal Toxicity, John Andrew Hitron

Theses and Dissertations--Toxicology and Cancer Biology

The reversible oxidation of cysteine by reactive oxygen species (ROS) is both a mechanism for cellular protein signaling as well as a cause of cellular injury and death through the generation of oxidative stress. The study of cysteine oxidation is complicated by the methodology currently available to isolate and enrich oxidized-cysteine containing proteins. We sought to simplify this process by reducing the time needed to process samples and reducing sample loss and contamination risk.

We accomplished this by eliminating precipitation steps needed for the protocol by (a) introducing an in-solution NEM-quenching step prior to reduction and (b) replacing soluble dithiothreitol …

Modulation Of Autophagy In Gbm Cells By Clinical Doses Of Gamma Radiation, Devron Ozgen

Graduate Research Theses & Dissertations

Glioblastoma (GBM) is a highly aggressive cancer that forms from the astrocytes of the brain. Poor prognosis and rather unsuccessful treatment regimens have left researchers and patients looking for new therapeutic strategies to combat this dreadful disease. Following tumor resection and radiotherapy combined with temozolomide, survival only remains around 15 months. Recurrence is very commonly seen, and urgently needs to be further understood. Autophagy is an important mechanism within cancer cells, which allows for cellular organelle/amino acid recycling and thus contributes to energy metabolism. Autophagy machinery has been shown to control cell death within cancer cells, leading to our interest …

Applications Of Cell-Derived Vesicles: From Single Molecule Studies To Drug Delivery, Faruk H. Moonschi

Theses and Dissertations--Chemistry

Single molecule studies can provide information of biological molecules which otherwise is lost in ensemble studies. A wide variety of fluorescence-based techniques are utilized for single molecule studies. While these tools have been widely applied for imaging soluble proteins, single molecule studies of transmembrane proteins are much more complicated. A primary reason for this is that, unlike membrane proteins, soluble proteins can be easily isolated from the cellular environment. One approach to isolate membrane proteins into single molecule level involves a very low label expression of the protein in cells. However, cells generate background fluorescence leading to a very low …

Investigating Autophagy, Extracellular Vesicles, And Glycobiology, Benjamin Cook

Master's Theses

Autophagy is a cellular process of degradation which degrades cellular materials in acidic organelles called lysosomes. Impairment of lysosomal degradation can lead to lysosomal storage disorders such as Parkinson's Disease. In Parkinson's Disease neurotoxic α-synuclein can spread from cell-to-cell via extracellular vesicles. We investigated the changes in extra vesicular glycans upon perturbation of autophagy using a class of glycan binding molecules termed lectins.

We found that upon induction of autophagy causes EV glycans to increase while others remained consistent. Additionally, we found that specific glycans are increased more than others. Conversely, when lysosomal degradation was impaired, we observed a decrease …

Appropriate Preconditioning Of The Uterine Endoplasmic Reticulum Stress Response Inhibits Preterm Labor, Judith Ann Ingles

Wayne State University Dissertations

Introduction: In this study, we are testing the overarching hypothesis that preconditioning the myometrial UPR allows for the maintenance of non-apoptotic CASP3 activity and thus sustains uterine quiescence. We have previously demonstrated that the pregnant uterus facilitates uterine quiescence through UPR mediated activation of non-apoptotic CASP3, yet the mechanism in which CASP3 utilizes to avoid its apoptotic cell fate is unresolved. There is a growing body of evidence including our own that demonstrates remote and direct preconditioning with minor stresses propagates cytoprotective mechanisms that allow for the avoidance of apoptotic cell death upon exposure to a subsequent more damaging stress, …

Evaluation And Adaptation Of Live-Cell Interferometry For Applications In Basic, Translational, And Clinical Research, Kevin A. Leslie

Theses and Dissertations

Cell mass is an important indicator of cell health and status. A diverse set of techniques have been developed to precisely measure the masses of single cells, with varying degrees of technical complexity and throughput. Here, the development of a non-invasive, label-free optical technique, termed Live-Cell Interferometry (LCI), is described. Several applications are presented, including an evaluation of LCI’s utility for assessing drug response heterogeneity in patient-derived melanoma lines and the measurement of CD3+ T cell kinetics during hematopoietic stem cell transplantation. The characterization of mast cells during degranulation, the measurement of viral reactivation kinetics in Kaposi’s Sarcoma, and drug …

Impact Of Aerobic Exercise On Monocyte Subset Receptor Expression And Macrophage Polarization, Anson M. Blanks

Theses and Dissertations

Atherosclerotic cardiovascular disease (CVD) is hallmarked by inflammatory immune activation, particularly by the induction of a response by monocytes. Classical (CD14⁺⁺CD16^-) are anti-inflammatory mediators under homeostatic conditions, while intermediate (CD14⁺⁺CD16⁺) and non-classical (CD14^LowCD16⁺⁺) monocytes promote inflammation following activation. Monocyte activation and functionality is dependent upon receptor expression and ligand production by a variety of cells, including monocytes. Alterations in the expression of surface receptors often have a direct impact upon monocyte function, such as the increased pro-inflammatory cytokine production in response to activation that accompanies elevated CD14 expression …

Non-Canonicaly Recruited Tcrαβcd8Αα Iels Recognize Microbial Antigens, Lukasz Wojciech, Edyta Szurek, Michal Kuczma, Anna Cebula, Wessam R. Elhefnawy, Maciej Pietrzak, Grzegorz Rempala, Leszek Ignatowicz

Computer Science Faculty Publications

In the gut, various subsets of intraepithelial T cells (IELs) respond to self or non-self-antigens derived from the body, diet, commensal and pathogenic microbiota. Dominant subset of IELs in the small intestine are TCRαβCD8αα⁺ cells, which are derived from immature thymocytes that express self-reactive TCRs. Although most of TCRαβCD8αα⁺ IELs are thymus-derived, their repertoire adapts to microbial flora. Here, using high throughput TCR sequencing we examined how clonal diversity of TCRαβCD8αα⁺ IELs changes upon exposure to commensal-derived antigens. We found that fraction of CD8αα⁺ IELs and CD4⁺ T cells express identical …

Charecterization Of The Innate Immune Response In The Rat Intervertebral Disc At The Tissue And Cellular Level, Matthew Guerreiro

Theses and Dissertations (Comprehensive)

Low back pain (LBP) affects up to 80% of the global population at some point during their life. The costs associated with LBP are estimated at $100 billion per year, putting significant strains on our healthcare system. As intervertebral disc (IVD) herniation is one of the major causes of LBP we decided to focus on herniation facilitated inflammatory responses, where damage caused by herniation triggers an innate immune response which likely results in pain, inflammation, and recruitment of specialized innate immune cells like macrophages to the affected area. Since the role of the inflammatory response is relatively uncharacterized in the …

A Physical Biology, The Electron Neutrino Mass, And The Role Of Quantum Mechanics In Nature, Maurice Goodman

Articles

In science we need to remove physics and earth science from the fundamental sciences and treat Biology as a fundamental natural (physical) science. Attempts to keep Biology autonomous because it is holistic, from the physical sciences, are just disguised anthropocentrism. Physical sciences have holistic features also. The autonomy of Biology is at odds with a holistic, integrated, science and is preventing progress in science. For example, every cell needs a 'global' communication system to keep order and stability with rapid information transfer across cellular scales. We have yet to figure out how this is achieved. Over the last 30 years, …