Cell and Developmental Biology Commons^™

The Transcriptional Regulation Of Flagellin-Induced Innate Protection Of The Cornea: Role Of Irf1 And Atf3, Gi Sang Yoon

Wayne State University Dissertations

Pre-exposure of the cornea to TLR5 ligand flagellin induces profound mucosal innate protection against infections by reprogramming gene expression. This study explored the flagellin-induced modifications of transcription factor expression and function, specifically of IRF1 and ATF3 in corneal epithelial cells to elucidate the transcriptional mechanisms underlying the protective function of flagellin on the cornea.

Initially we used Superarray to screen for transcription factors and identified Interferon Regulatory Factor (IRF) 1 and Activating Transcription Factor (ATF) 3 as the most drastically affected genes by flagellin pretreatment in P. aeruginosa challenged human corneal epithelial cells (CEC). However, flagellin pretreatment had opposite effects …

Keratin Gene Expression Profiles After Digit Amputation In C57bl/6 Vs. Regenerative Mrl Mice Imply An Early Regenerative Keratinocyte Activated-Like State, Chia-Ho Cheng, John Leferovich, Xiangming Zhang, Khamilia Bedelbaeva, Dmitri Gourevitch, Cathy J. Hatcher, Craig T. Basson, Ellen Heber-Katz, Kenneth A. Marx

PCOM Scholarly Works

Mouse strains C57BL/6 (B6) and MRL were studied by whole mouse genome chip microarray analyses of RNA isolated from amputation sites at different times pre-and postamputation at the midsecond phalange of the middle digit. Many keratin genes were highly differentially expressed. All keratin genes were placed into three temporal response classes determined by injury/preinjury ratios. One class, containing only Krt6 and Krt16, were uniquely expressed relative to the other two classes and exhibited different temporal responses in MRL vs. B6. Immunohistochemical staining for Krt6 and Krt16 in tissue sections, including normal digit, flank skin, and small intestine, and from normal …

A Physical Sciences Network Characterization Of Non-Tumorigenic And Metastatic Cells, Abigail Hielscher, D. Wirtz, Et Al.

PCOM Scholarly Works

To investigate the transition from non-cancerous to metastatic from a physical sciences perspective, the Physical Sciences-Oncology Centers (PS-OC) Network performed molecular and biophysical comparative studies of the non-tumorigenic MCF-10A and metastatic MDA-MB-231 breast epithelial cell lines, commonly used as models of cancer metastasis. Experiments were performed in 20 laboratories from 12 PS-OCs. Each laboratory was supplied with identical aliquots and common reagents and culture protocols. Analyses of these measurements revealed dramatic differences in their mechanics, migration, adhesion, oxygen response, and proteomic profiles. Model-based multi-omics approaches identified key differences between these cells' regulatory networks involved in morphology and survival. These results …

Matrigel Modulates A Stem Cell Phenotype And Promotes Tumor Cell Formation In A Mantle Cell Lymphoma Cell Line, Abigail Hielscher, Timothy Mcguire, Dennis Weisenburger, John Graham Sharp

PCOM Scholarly Works

Tumors may be maintained by subpopulations of cells possessing stem cell-like properties. We evaluated the stem cell-like and tumor-forming properties of side population (SP) and CD133+/ CD44+ cells in Granta 519, a human mantle cell lymphoma cell line. The in-vitro Cobblestone Area Forming Cell (CAFC) assay, designed to detect stem and progenitor cells, revealed that SP cells contained the greatest proportion of stem cell-like cells. The addition of Matrigel to CAFC assays of SP and non-SP cells both increased their respective stem cell frequencies in comparison to those cultures without Matrigel, and additionally resulted in observed stem cell frequencies which …

Hypoxia Affects The Structure Of Breast Cancer Cell Derived Matrix To Support Angiogenic Responses Of Endothelial Cells, Abigail Hielscher, Connie Qiu, Josh Porterfield, Quinton Smith, Sharon Gerecht

PCOM Scholarly Works

Hypoxia, a common feature of the tumor environment and participant in tumor progression, is known to alter gene and protein expression of several Extracellular Matrix (ECM) proteins, many of which have roles in angiogenesis. Previously, we reported that ECM deposited from co cultures of Neonatal Fibroblasts (NuFF) with breast cancer cells, supported 3-dimensional vascular morphogenesis. Here, we sought to characterize the hypoxic ECM and to identify whether the deposited ECM induce angiogenic responses in Endothelial Cells (ECs). NuFF and MDAMB 231 breast cancer cells were co-cultured, subjected to alternating cycles of 24 hours of 1% (hypoxia) and 21% (atmospheric) oxygen …

Characterization Of G-Patch Motif Contribution To Prp43 Function In The Pre-Messenger Rna Splicing And Ribosomal Rna Biogenesis Pathways, Daipayan Banerjee

Theses and Dissertations--Biology

The DExD/H-box protein Prp43 is essential for two biological processes: nucleoplasmic pre-mRNA splicing and nucleolar rRNA maturation. The biological basis for the temporal and spatial regulation of Prp43 remains elusive. The Spp382/Ntr1, Sqs1/Pfa1 and Pxr1/Gno1 G-patch proteins bind to and activate the Prp43 DExD/H box-helicase in pre-mRNA splicing (Spp382) and rRNA processing (Sqs1, Pxr1). These Prp43-interacting proteins each contain the G-patch domain, a conserved sequence of ~48 amino acids that includes 6 highly conserved glycine (G) residues. Five annotated G-patch proteins in baker’s yeast (i.e., Spp382, Pxr1, Spp2, Sqs1 and Ylr271) and with the possible exception of the uncharacterized Ylr271 …

Manipulation Of Cellular Processing Bodies And Their Constituents By Viruses, Asit K. Pattnaik, Phat X. Dinh

School of Veterinary and Biomedical Sciences: Faculty Publications

The processing bodies (PBs) are a form of cytoplasmic aggregates that house the cellular RNA decay machinery as well as many RNA-binding proteins and mRNAs. The PBs are constitutively present in eukaryotic cells and are involved in maintaining cellular homeostasis by regulating RNA metabolism, cell signaling, and survival. Virus infections result in modification of the PBs and their constituents. Many viruses induce compositionally altered PBs, while many others use specific components of the PBs for their replication. PB constituents are also known to restrict virus replication by a variety of mechanisms. Further, continuing studies in this rapidly emerging field of …

Deep Sequencing And Genome-Wide Analysis Reveals The Expansion Of Microrna Genes In The Gall Midge Mayetiola Destructor, Chitvan Khajuria, Christie E. Williams, Mustapha El Bouhssini, R. Jeff Whitworth, Stephen Richards, Jeffrey J. Stuart, Ming-Shun Chen

School of Veterinary and Biomedical Sciences: Faculty Publications

Background: MicroRNAs (miRNAs) are small non-coding RNAs that play critical roles in regulating post transcriptional gene expression. Gall midges encompass a large group of insects that are of economic importance and also possess fascinating biological traits. The gall midge Mayetiola destructor, commonly known as the Hessian fly, is a destructive pest of wheat and model organism for studying gall midge biology and insect – host plant interactions.

Results: In this study, we systematically analyzed miRNAs from the Hessian fly. Deep-sequencing a Hessian fly larval transcriptome led to the identification of 89 miRNA species that are either identical or very …

Freeze Tolerance Conferred By Transgenic Expression Of Late Embryogenesis Abundant Protein In Insect Cells, Alyssa N. Walser

Masters Theses

Anhydrobiotic organisms such as the brine shrimp Artemia franciscana have developed strategies to survive and thrive under conditions of limited water availability. One strategy is the accumulation of late embryo genesis abundant (LEA) proteins. Cells of Drosophila melanogaster (Kc167), which had previously been genetically modified to express a group 1 LEA protein (LEA1.3) found in A. franciscana, were used to test the impact of water stress due to freezing to -80°C. Two different freezing devices were used to determine the impact of the freezing protocol on the viability of cells after freezing and thawing. The first method used a 12-well …

Molecular Mechanisms Involved In Anhydrobiosis Of Insect Cells, Diyagama Arachchi Ralalage Dilini Sewwandi Samarajeewa

Masters Theses

Animals possessing tolerance to extreme water stress are termed anhydrobiotes. Many desiccation tolerant organisms respond to water stress by intracellular accumulation of selected sugars such as trehalose and larger macromolecules such as Late Embryogenesis Abundant (LEA) proteins and thereby maintain the cell viability. Evidence indicates that the presence of trehalose and Late Embryogenesis Abundant (LEA) proteins may work synergistically to confer cellular protection during drying in eukaryotic cells. We evaluated any increase in cellular desiccation tolerance by expressing different LEA proteins in a non-desiccation tolerant insect cell line Drosophila melanogaster (fruit fly) (Kc167 cells) in the presence of trehalose.

Transgenic …

Two Modes Of Cell Death Caused By Exposure To Nanosecond Pulsed Electric Field, Olga N. Pakhomova, Betsy W. Gregory, Iurii Semenov, Andrei G. Pakhomov

Bioelectrics Publications

High-amplitude electric pulses of nanosecond duration, also known as nanosecond pulsed electric field (nsPEF), are a novel modality with promising applications for cell stimulation and tissue ablation. However, key mechanisms responsible for the cytotoxicity of nsPEF have not been established. We show that the principal cause of cell death induced by 60- or 300-ns pulses in U937 cells is the loss of the plasma membrane integrity (‘‘nanoelectroporation’’), leading to water uptake, cell swelling, and eventual membrane rupture. Most of this early necrotic death occurs within 1–2 hr after nsPEF exposure. The uptake of water is driven by the presence of …

Cell Responses Without Receptors And Ligands, Using Nanosecond Pulsed Electric Fields (Nspefs), Stephen J. Beebe

Bioelectrics Publications

No abstract provided.

Late Developing Mammary Tumors And Hyperplasia Induced By A Low-Oncogenic Variant Of Mouse Mammary Tumor Virus (Mmtv) Express Genes Identical To Those Induced By Canonical Mmtv, Robert D. Bruno

School of Medical Diagnostics & Translational Sciences Publications

Background: The canonical milk-transmitted mouse mammary tumor virus (MMTV) of C3H mice (C3H-MMTV) rapidly induces tumors in 90% of infected animals by 8 months of age. Pro-viral insertions of C3H-MMTV into genomic DNA results in the overexpression of common core insertion site (CIS) genes, including Wnt1/10b, Rspo2, and Fgf3. Conversely, infection by either the endogenous Mtv-1 virus (in C3Hf) or the exogenous nodule-inducing virus (NIV) (in Balb/c NIV) induces premalignant mammary lesions and tumors with reduced incidence and longer latency than C3H-MMTV. Here, we asked whether Mtv-1/NIV affected the expression of core CIS genes.

Findings: We confirmed the presence of …

Embryonic Stem Cells Are Redirected To Non-Tumorigenic Epithelial Cell Fate By Interaction With The Mammary Microenvironment, Corinne A. Boulanger, Robert D. Bruno, David L. Mack, Monica Gonzales, Nadia P. Castro, David S. Salomon, Gilbert H. Smith

School of Medical Diagnostics & Translational Sciences Publications

Experiments were conducted to redirect mouse Embryonic Stem (ES) cells from a tumorigenic phenotype to a normal mammary epithelial phenotype in vivo. Mixing LacZ-labeled ES cells with normal mouse mammary epithelial cells at ratios of 1:5 and 1:50 in phosphate buffered saline and immediately inoculating them into epithelium-divested mammary fat pads of immune-compromised mice accomplished this. Our results indicate that tumorigenesis occurs only when normal mammary ductal growth is not achieved in the inoculated fat pads. When normal mammary gland growth occurs, we find ES cells (LacZ+) progeny interspersed with normal mammary cell progeny in the mammary epithelial structures. We …

The Bio-Nano-Interface In Predicting Nanoparticle Fate And Behaviour In Living Organisms: Towards Grouping And Categorising Nanomaterials And Ensuring Nanosafety By Design, Hugh Byrne, Arti Ahluwalia, Diana Boraschi,, Bengt Fadeel, Peter Gehr, Arno C. Gutleb, Michaela Kendall, Manthos Papadopoulos, Iseult Lynch

Articles

In biological media, nanoparticles acquire a coating of biomolecules (proteins, lipids, polysaccharides) from their surroundings, which reduces their surface energy and confers a biological identity to the particles. This adsorbed layer is the interface between the nanomaterial and living systems and therefore plays a significant role in determining the fate and behaviour of the nanoparticles. This review summarises the state of the art in terms of understanding the bio-nano interface and provides direction for potential future research directions and some recommendations for future priorities and strategies to support the safe implementation of nanotechnologies. The central premise is that nanomaterials must …

Regulation Of The Glycine Transporter 1 (Glyt1) By Pkca-Dependent Ubiquitination, Susana Barrera

Open Access Theses & Dissertations

Glycine is one of the major inhibitory neurotransmitters in the central nervous system. It is implicated in the regulation of motor-sensory function such as; pain perception, reflex responses, and is essential for early development of the CNS. In addition to its inhibitory actions, it acts as an obligatory co-agonist for the activation of glutamatergic NMDARs, which are of pivotal importance in the process of learning and memory, its dysfunction leads to several neurodegenerative diseases. Two high affinity Na+/Cl- dependent transporters GlyT1 and GlyT2 tightly regulate the availability of glycine present at the synapse for the activation of NMDARs and GlyRs. …

Comparison Of Ectoparasitic Mite Loads Between Gonochoristic (Aspidoscelis Marmorata) And Parthenogenic (A. Tesselata) Syntopic Whiptail Lizards (Teiidae) From The Northern Chihuahuan Desert Of Trans-Pecos, Texas, William Dawson Lukefahr

Open Access Theses & Dissertations

The objective of this study was to compare the ectoparasitic loads between two syntopic whiptail lizard species (Teiidae) that differ in their reproductive modes; Aspidoscelis marmorata (bisexual) and A. tesselata (unisexual). The study site was located on Indio Mountains Research Station (IMRS) in a Chihuahuan Desert Scrub landscape in Hudspeth County, Texas. Previous studies conducted at IMRS showed the two species having similar diets, morphology, and reproductive characteristics, but fluctuated in nematode loads. No comparative study of ecotoparasitic mites (Eutrombicula alfreddugesi) between the two species has ever been done.

Aspidoscelis marmorata (n = 141) and A. tesselata (n = 144) …

Novel Copper (Ii) Mediated Transfection Nano-Systems And Biological Properties Of Pyridinium Surfactants, Sarit Pal

Open Access Theses & Dissertations

Efficacy of gene therapy is relied upon the improvement of gene delivery vectors into mammalian cells. Although viral-based polynucleotide carriers are still considered as the most common method for gene delivery but the cost effectiveness, nonspecific lethal immune response has evoked the renaissance of non-viral synthetic delivery systems. An initial discovery by Felgnar et al showed Cationic Liposomes (CLs) (having an overall positive charge) when mixed with DNA to form CL-DNA complexes can enhance the transfer of polynucleotides into cells followed by its expression. Presently, a flurry of experimental works is taking place for the better understanding of structural characterization …

Effects Of Acute Alcohol Exposure On Post Burn Intestinal Immunity: Role Of Il-23, Juan L. Rendon

Dissertations

More than one million burn injuries are reported yearly within the United States. These injuries result in approximately 500,000 emergency room visits and 40,000 hospitalizations annually. Greater than 50% of these injuries occur under the influence of alcohol/ethanol (EtOH) intoxication. Burn victims who sustain injury under the influence of EtOH exhibit significantly higher rates of morbidity and mortality than patients without EtOH exposure at the time of injury. Several lines of evidence suggest that gut pathogens and/or their products may play a role in the development of sepsis and multiple organ failure reported in burn and trauma patients. In line …

Facilitation Of Electroporative Drug Uptake And Cell Killing By Electrosensitization, Olga N. Pakhomova, Betsy W. Gregory, Andrei G. Pakhomov

Bioelectrics Publications

Cell permeabilization by electric pulses (EP), or electroporation, is widely used for intracellular delivery of drugs and plasmids, as well as for tumour and tissue ablation. We found that cells pre-treated with 100-mus EP develop delayed hypersensitivity to subsequent EP applications. Sensitizing B16 and CHO cells by splitting a single train of eight 100-mus EP into two trains of four EP each (with 5-min. interval) decreased the LD(50) 1.5-2 times. Sensitization profoundly enhanced the electroporation-assisted uptake of bleomycin, a cell-impermeable cytotoxic agent accepted for killing tumours by electrochemotherapy. EP exposures that were not lethal per se caused cell death in …