Medical Cell Biology Commons^™

Structural And Functional Interactions Between Bro1 Domain Of Human Alix Protein And Nucleocapsid Packaging Rna Complex From Hiv, Scott Gross

Graduate School of Biomedical Sciences Theses and Dissertations

A virus is only as powerful as its ability to spread. Enveloped retroviruses, namely HIV-1, use exocytosis pathways that normal host cells use to release particles from the plasma membrane. The main pathways of interest in this study are the Endosomal Sorting Complex Required for Transport (ESCRT) and adjacent ALIX pathways. The ESCRT pathway is especially important for degradation of receptor/cargo complexes that form Multi-Vesicular Bodies (MVBs). Currently, there is no known therapy that targets this endosomal pathway, which would prevent the spread of the virus to other cells. The virus has adapted to jump from pathway to pathway when …

Multilevel Deregulation Of Survival Mechanisms In Npm-Alk+ T-Cell Lymphoma, Deeksha Vishwamitra

Dissertations & Theses (Open Access)

The anaplastic lymphoma kinase (ALK) is a single chain transmembrane receptor tyrosine kinase that belongs to the insulin receptor superfamily. Other members of this superfamily include the insulin receptor (IR), type I insulin-like growth factor receptor (IGF-IR), and the leukocyte tyrosine kinase. The common structural finding among these tyrosine kinases is the YXXXYY motif present within their respective tyrosine kinase domains. Binding of its ligands causes ALK receptor homodimerization and protein kinase activation. ALK has been previously shown to play a significant role during early developmental stages. In human embryos, the expression of ALK is mainly seen in …

Structural-Based Investigation Of Fak And P130cas In Focal Adhesion, Chi Zhang

Theses and Dissertations (ETD)

Cell migration is an integrated multistep process accompanying us throughout life, from birth to death. It contributes to a variety of biological activities and has been implicated in many pathological conditions, including cancer, primary immunodeficiency diseases, vascular diseases, and mental retardation. Cell migration has been considered to be a three-step cyclic process including polarization, protrusion and retraction. The initiation and extension of the protrusion is driven by actin polymerization, and is stabilized by attaching to the extracellular matrix (ECM) or adjacent cells. The cell-ECM attachment sites are named focal adhesions (FAs), which are dynamic, large protein complexes containing more than …

Diabetic Bone Marrow & Stem Cell Dysfunction, Meghan A. Piccinin

Electronic Thesis and Dissertation Repository

ii Abstract Abstract Defects in the proliferation, differentiation, and activity of bone marrow (BM)-derived vasculogenic/vascular stem cells (VSCs) have been observed in diabetes and contribute to the development of vascular complications. Diabetes leads to enhanced bone marrow adipogenesis, altering the composition of the BM stem cell (SC) niche and potentially disrupting the normal functioning of resident VSCs. Here, I establish that adipocytes have a negative influence on SC survival in culture. I also show that adipocytes and osteoblasts are responsible for the creation of distinct extracellular microenvironments, with unique expression patterns of several pro- and anti-angiogenic factors with known effects …

Timp3 Regulation Of Macrophage Activation And Apoptosis, Michael S. Brock

Electronic Thesis and Dissertation Repository

Acute respiratory distress syndrome (ARDS) is a lung disease involving profound inflammation. Origins of persistent inflammation in select cases of ARDS are poorly understood, and we propose persistent inflammatory macrophages may be one of its mechanisms. Macrophages polarize to either promote inflammation, or suppress inflammation. Tissue inhibitor of metalloproteinases 3 (TIMP3) reduces the pro-inflammatory polarization in macrophages. Additionally, studies have shown TIMP3 promotes apoptosis, and its absence delays recovery from bleomycin-induced lung injury.

We hypothesize that TIMP3 promotes apoptosis of murine macrophages through inhibition of metalloproteinase activity and stabilization of FAS on the cell surface. Pro-inflammatory Timp3^-/- bone marrow-derived …

One-Step Generation Of Multiple Transgenic Mouse Lines Using An Improved Pronuclear Injection-Based Targeted Transgenesis (I-Pitt)., Masato Ohtsuka, Hiromi Miura, Keiji Mochida, Michiko Hirose, Ayumi Hasegawa, Atsuo Ogura, Ryuta Mizutani, Minoru Kimura, Ayako Isotani, Masahito Ikawa, Masahiro Sato, Channabasavaiah B. Gurumurthy

Journal Articles: Genetics, Cell Biology & Anatomy

BACKGROUND: The pronuclear injection (PI) is the simplest and widely used method to generate transgenic (Tg) mice. Unfortunately, PI-based Tg mice show uncertain transgene expression due to random transgene insertion in the genome, usually with multiple copies. Thus, typically at least three or more Tg lines are produced by injecting over 200 zygotes and the best line/s among them are selected through laborious screening steps. Recently, we developed technologies using Cre-loxP system that allow targeted insertion of single-copy transgene into a predetermined locus through PI. We termed the method as PI-based Targeted Transgenesis (PITT). A similar method using PhiC31-attP/B system …

Lipid And Protein Co-Regulation Of Pi3k Effectors Akt And Itk In Lymphocytes, Xinxin Wang, Leonard B. Hills, Yina H. Huang

Dartmouth Scholarship

The phosphoinositide 3-kinase (PI 3-kinase, PI3K) pathway transduces signals critical for lymphocyte function. PI3K generates the phospholipid PIP3 at the plasma membrane to recruit proteins that contain pleckstrin homology (PH) domains - a conserved domain found in hundreds of mammalian proteins. PH domain-PIP3 interactions allow for rapid signal propagation and confer a spatial component to these signals. The kinases Akt and Itk are key PI3K effectors that bind PIP3 via their PH domains and mediate vital processes - such as survival, activation, and differentiation - in lymphocytes. Here, we review the roles and regulation of PI3K signaling in lymphocytes with …

Insertion Of Sequences At The Original Provirus Integration Site Of Mouse Rosa26 Locus Using The Crispr/Cas9 System., Rolen M. Quadros, Donald W. Harms, Masato Ohtsuka, Channabasavaiah B. Gurumurthy

Journal Articles: Genetics, Cell Biology & Anatomy

Targeted transgenic mouse models, where an exogenous gene is inserted into a specified genomic locus to achieve its stable and reliable expression, have been widely used in biomedical research. However, the available methodologies for targeted insertion of sequences require many laborious steps that involve the use of embryonic stem (ES) cells. We recently developed Pronuclear Injection-based Targeted Transgenesis (PITT), a method that uses a recombinase-mediated cassette exchange (RMCE) to enable insertion of sequences at a predetermined genomic locus, such as ROSA26. The PITT technique uses fertilized eggs (instead of ES cells) collected from 'seed mice' that contain the RMCE landing …

Natural Killer Cells In Patients With Severe Chronic Fatigue Syndrome, Ekua Brenu, Sharni Hardcastle, Gunn Atkinson, Mieke Van Driel, Sanne Kreijkamp-Kaspers, Kevin Ashton, Don Staines, Sonja Marshall-Gradisnik

Kevin Ashton

Maintenance of health and physiological homeostasis is a synergistic process involving tight regulation of proteins, transcription factors and other molecular processes. The immune system consists of innate and adaptive immune cells that are required to sustain immunity. The presence of pathogens and tumour cells activates innate immune cells, in particular Natural Killer (NK) cells. Stochastic expression of NK receptors activates either inhibitory or activating signals and results in cytokine production and activation of pathways that result in apoptosis of target cells. Thus, NK cells are a necessary component of the immunological process and aberrations in their functional processes, including equivocal …

Generation Of A Retinoblastoma (Rb)1-Inducible Dominant-Negative (Dn) Mouse Model., Shikha Tarang, Songila M.S.R Doi, Channabasavaiah B. Gurumurthy, Donald W. Harms, Rolen M. Quadros, Sonia M. Rocha-Sanchez

Journal Articles: Genetics, Cell Biology & Anatomy

Retinoblastoma 1 (Rb1) is an essential gene regulating cellular proliferation, differentiation, and homeostasis. To exert these functions, Rb1 is recruited and physically interacts with a growing variety of signaling pathways. While Rb1 does not appear to be ubiquitously expressed, its expression has been confirmed in a variety of hematopoietic and neuronal-derived cells, including the inner ear hair cells (HCs). Studies in transgenic mice demonstrate that complete germline or conditional Rb1 deletion leads to abnormal cell proliferation, followed by massive apoptosis; making it difficult to fully address Rb1's biochemical activities. To overcome these limitations, we developed a tetracycline-inducible TetO-CB-myc6-Rb1 (CBRb) mouse …

A Comparison Of Four Electrical Stimulation Types On Staphylococcus Aureus Growth In Vitro, Harold Merriman, Chris Hegyi, Cheryl Albright-Overton, John Carlos, Robert Putnam, Janet Mulcare

Harold L. Merriman

We evaluated the efficacy of common electrical stimulation (ES) types on bacterial growth in vitro using clinically relevant conditions. Four types of ES-continuous microamperage direct current (µADC), high-voltage pulsed current (HVPC), low-voltage monophasic milliamperage pulsed current (LVMmAPC), and low-voltage biphasic milliamperage pulsed current (LVBmAPC)-were each applied to a separate set of culture plates containing Staphylococcus aureus for 1 h at 37 °C on 3 consecutive days. After ES treatment, the zone of inhibition surrounding each electrode was measured. Zone of inhibition measurements showed a significant inhibitory effect for continuous µADC and HVPC (p < 0.05), but not for LVMmAPC and LVBmAPC. …

Iron Alters Cell Survival In A Mitochondria-Dependent Pathway In Ovarian Cancer Cells., Edward Haller

Edward Haller

ABSTRACT The role of iron in the development of cancer remains unclear. We previously reported that iron reduces cell survival in a Ras/mitogen-activated protein kinase (MAPK)-dependent manner in ovarian cells; however, the underlying downstream pathway leading to reduced survival was unclear. Although levels of intracellular iron, ferritin/CD71 protein and reactive oxygen species did not correlate with iron-induced cell survival changes, we identified mitochondrial damage (via TEM) and reduced expression of outer mitochondrial membrane proteins (translocase of outer membrane: TOM20 and TOM70) in cell lines sensitive to iron. Interestingly, Ru360 (an inhibitor of the mitochondrial calcium uniporter) reversed mitochondrial changes and …

Mitochondrial Ca(2+) Uptake By The Voltage-Dependent Anion Channel 2 Regulates Cardiac Rhythmicity., Hirohito Shimizu, Johann Schredelseker, Jie Huang, Kui Lu, Shamim Naghdi, Fei Lu, Sarah Franklin, Hannah Dg Fiji, Kevin Wang, Huanqi Zhu, Cheng Tian, Billy Lin, Haruko Nakano, Amy Ehrlich, Junichi Nakai, Adam Z Stieg, James K Gimzewski, Atsushi Nakano, Joshua I. Goldhaber, Thomas M. Vondriska, György Hajnóczky, Ohyun Kwon, Jau-Nian Chen

Department of Pathology, Anatomy, and Cell Biology Faculty Papers

Tightly regulated Ca(2+) homeostasis is a prerequisite for proper cardiac function. To dissect the regulatory network of cardiac Ca(2+) handling, we performed a chemical suppressor screen on zebrafish tremblor embryos, which suffer from Ca(2+) extrusion defects. Efsevin was identified based on its potent activity to restore coordinated contractions in tremblor. We show that efsevin binds to VDAC2, potentiates mitochondrial Ca(2+) uptake and accelerates the transfer of Ca(2+) from intracellular stores into mitochondria. In cardiomyocytes, efsevin restricts the temporal and spatial boundaries of Ca(2+) sparks and thereby inhibits Ca(2+) overload-induced erratic Ca(2+) waves and irregular contractions. We further show that overexpression …

In Vivo Cigarette Smoke Exposure Decreases Ccl20, Slpi, And Bd-1 Secretion By Human Primary Nasal Epithelial Cells, James Jukosky, Benoit J. Gosselin, Leah Foley, Tenzin Dechen, Steven Fiering, Mardi A. Crane-Godreau

Dartmouth Scholarship

Smokers and individuals exposed to second-hand cigarette smoke have a higher risk of developing chronic sinus and bronchial infections. This suggests that cigarette smoke (CS) has adverse effects on immune defenses against pathogens. Epithelial cells are important in airway innate immunity and are the first line of defense against infection. Airway epithelial cells not only form a physical barrier but also respond to the presence of microbes by secreting antimicrobials, cytokines, and chemokines. These molecules can lyse infectious microorganisms and/or provide signals critical to the initiation of adaptive immune responses. We examined the effects of CS on antimicrobial secretions of …

The Cell's Antenna And Bending With The Flow, Surya M. Nauli, Kimberly F. Atkinson, Sarmed H. Kathem

Pharmacy Faculty Articles and Research

Polycystic kidney disease (PKD) is the most common life-threatening genetic disease worldwide – affecting about 12.5 million people. Patients with the condition have multiple fluid-filled cysts in their kidneys that lead to a massive enlargement of the organ and gradual worsening of its function, as well as complete failure in many cases. The lack of available treatment means patients may eventually require regular dialysis or a kidney transplant.

Towards A Structural Understanding Of Spore Germination In Clostridium Difficile, Chloe M. Adams

Graduate College Dissertations and Theses

Clostridium difficile is a Gram-positive bacterium that causes a toxin-mediated disease, typically in individuals whose normal intestinal flora has been compromised by antibiotic therapy. C. difficile is naturally resistant to many antibiotics and produces spores that can withstand harsh environmental conditions and many disinfectants, making the infection difficult to clear and easy to spread. The infection begins when spores from the environment are ingested and germinate upon exposure to taurocholate and glycine in the digestive tract. This germination process is required to initiate infection and thus represents a good target for the development of novel therapeutics. Although spore germination is …

Pancreas: Do All Roads Lead To Mitochondria?, Amit Mukherji, Omobola Onikoyi, Vasudeva G. Kamath

Touro College of Osteopathic Medicine (Middletown) Publications and Research

Over several millions of years of evolution, mitochondria have transformed into specialized organelles. Today, they cannot live outside the cell nor can the host cell live without them, resulting in a symbiotic relationship. Richard Altmann, in 1894, documented them as cell organelles and called them “bioblasts”. Later, the term “mitochondria” itself was coined by Carl Benda in 1898. Ever since these findings, we in the field of medicine have learned a lot about this tiny organelle, but numerous aspects continue to be discovered. In this article, we will review the significance of this organelle in terms of pancreatic dysfunctions.

Improved Accuracy Of Surgical Pathology Accessioning, Coding, And Billing Practices After Self-Evaluation, Allison F. Goldberg, Md

Department of Pathology, Anatomy, and Cell Biology Resident's Posters

INTRODUCTION

Accurate accessioning, coding, and billing of specimens are vital, but often disregarded, aspects of a surgical pathologist’s job. This study had two aims related to those aspects. First, to evaluate the quality of our systems based practices in order to identify areas for improvement. Second, to assess if changes made based on our self-evaluation improved accuracy.

Tumor Metabolism In The Microenvironment Of Nodal Metastasis In Head And Neck Squamous Cell Carcinoma, Mehri Mollaee, Md, Patrick T. Tassone, Md, David Cognetti, Md, Adam J. Luginbuhl, Md, Joanie Johnson, Ubaldo E. Martinez-Outshoorn, Md, Tingting Zhan, Joseph M. Curry, Md, Madalina Tuluc, Md

Department of Pathology, Anatomy, and Cell Biology Resident's Posters

ABSTRACT

Introduction: Monocarboxylate transporter 4 (MCT4) is a cell membrane transporter of lactate. MCT4 is a tumor-specific marker of oxidative stress, glycolysis and hypoxia in tumor stromal cells. We investigated HPV positive and negative tumors with regional metastases to cervical lymph nodes (LN) to study how the metastatic tumor cells interact with their microenvironment. By selecting cancers with extracapsular extension (ECE), we intended to evaluate the interaction between metastases and the surrounding extranodal tissue.

Methods:

Clinical data were collected from 24 advanced stage oropharyngeal squamous cell carcinoma (OPSCC) patients with neck LN metastasis. All patients presented with at least N1 …

Diagnostic Value Of Endoscopic Ultrasound-Guided Fine Needle Aspiration Of Intra-Abdominal Lymph Nodes In Patients With Concurrent Biopsy Of Intra-Abdominal Tumors, Allison F. Goldberg, Md, Charalambos C. Solomides, Md, Marluce Bibbo, Md, Rossitza Draganova-Tacheva, Md

Department of Pathology, Anatomy, and Cell Biology Resident's Posters

BACKGROUND

• Diagnostic endoscopic biopsy of intra-abdominal lesions may be performed prior to surgical resections.

• Endoscopic ultrasound-guided fine needle aspiration of intra-abdominal lymph nodes may also be performed to:

• Yield more information

• Allow for more accurate staging

• We evaluate the additional staging information concurrent fine needle aspiration of intra-abdominal lymph nodes provided

DESIGN

• We included all patients at our institution from January 1, 2000 to March 30, 2015 who, during the same endoscopic procedure, had:

• Endoscopic ultrasound guided fine needle aspiration of an intra-abdominal lymph node AND

• Endoscopic ultrasound guided fine needle aspiration …