Cell Biology Commons^™

Peptidoglycan Recycling In Gram-Positive Bacteria Is Crucial For Survival In Stationary Phase, Marina Borisova, Rosmarie Gaupp, Amanda Duckworth, Alexander Schneider, Désirée Dalügge, Maraike Mühleck, Denise Deubel, Sandra Unsleber, Wenqi Yu, Günther Muth, Markus Bischoff, Friedrich Götz, Christoph Mayer

Molecular Biosciences Faculty Publications

Peptidoglycan recycling is a metabolic process by which Gram-negative bacteria reutilize up to half of their cell wall within one generation during vegetative growth. Whether peptidoglycan recycling also occurs in Gram-positive bacteria has so far remained unclear. We show here that three Gram-positive model organisms, Staphylococcus aureus, Bacillus subtilis, and Streptomyces coelicolor, all recycle the sugar N-acetylmuramic acid (MurNAc) of their peptidoglycan during growth in rich medium. They possess MurNAc-6-phosphate (MurNAc-6P) etherase (MurQ in E. coli) enzymes, which are responsible for the intracellular conversion of MurNAc-6P to N-acetylglucosamine-6-phosphate and d-lactate. By applying mass spectrometry, we observed accumulation of MurNAc-6P in …

Mir146a-Mediated Targeting Of Fancm During Inflammation Compromises Genome Integrity, Devakumar Sundaravinayagam, Hye R. Kim, Ting T. Wu, Hyun H. Kim, Hyun-Seo Lee, Semo Jun, Jeong-Heon Cha, Younghoon Kee, Ho J. You, Jung-Hee Lee

Molecular Biosciences Faculty Publications

Inflammation is a potent inducer of tumorigenesis. Increased DNA damage or loss of genome integrity is thought to be one of the mechanisms linking inflammation and cancer development. It has been suggested that NF-κB-induced microRNA-146 (miR146a) may be a mediator of the inflammatory response. Based on our initial observation that miR146a overexpression strongly increases DNA damage, we investigated its potential role as a modulator of DNA repair. Here, we demonstrate that FANCM, a component in the Fanconi Anemia pathway, is a novel target of miR146a. miR146a suppressed FANCM expression by directly binding to the 3’ untranslated region of the gene. …

The Cytotoxic Synergy Of Nanosecond Electric Pulses And Low Temperature Leads To Apoptosis, Claudia Muratori, Andrei G. Pakhomov, Elena C. Gianulis, Sarah D. Jensen, Olga N. Pakhomova

Bioelectrics Publications

Electroporation by nanosecond electric pulses (nsEP) is an emerging modality for tumor ablation. Here we show the efficient induction of apoptosis even by a non-toxic nsEP exposure when it is followed by a 30-min chilling on ice. This chilling itself had no impact on the survival of U-937 or HPAF-II cells, but caused more than 75% lethality in nsEP-treated cells (300 ns, 1.8-7 kV/cm, 50-700 pulses). The cell death was largely delayed by 5-23 hr and was accompanied by a 5-fold activation of caspase 3/7 (compared to nsEP without chilling) and more than 60% cleavage of poly-ADP ribose polymerase (compared …

Cytosolic Dna Sensor Upregulation Accompanies Dna Electrotransfer In B16.F10 Melanoma Cells, Katarina Znidar, Masa Bosnjak, Maja Cemazar, Loree C. Heller

Bioelectrics Publications

In several preclinical tumor models, antitumor effects occur after intratumoral electroporation, also known as electrotransfer, of plasmid DNA devoid of a therapeutic gene. In mouse melanomas, these effects are preceded by significant elevation of several proinflammatory cytokines. These observations implicate the binding and activation of intracellular DNA-specific pattern recognition receptors or DNA sensors in response to DNA electrotransfer. In tumors, IFN β mRNA and protein levels significantly increased. The mRNAs of several DNA sensors were detected, and DAI, DDX60, and p204 tended to be upregulated. These effects were accompanied with reduced tumor growth and increased tumor necrosis. In B16. F10 …

Expression Of Artemia Lea Proteins In Drosophila Melanogaster Cells Using Multicistronic Vector Constructs, Kazi Nazrul Islam

Masters Theses

In nature several organisms exhibit anhydrobiosis, the outstanding feature to survive in extreme desiccation by entering into a state of dormancy known as diapause. The cyst of the brine shrimp Artemia franciscana shows anhydrobiosis by entering into a diapause phase. Previous studies showed a correlation between anhydrobiosis and expression of highly hydrophilic polypeptides termed late embryogenesis abundant (LEA) proteins. However, the precise molecular mechanisms of LEA proteins are still unknown. The presence of multiple LEA proteins in Artemia suggests that some of them might work together. Here, I aimed to express different combination of two LEA proteins from Artemia franciscana …

The Roles Of Human Cytomegalovirus Tegument Proteins Pul48 And Pul103 During Lytic Infection, Daniel Angel Ortiz

Wayne State University Dissertations

THE ROLES OF HUMAN CYTOMEGALOVIRUS TEGUMENT PROTEINS pUL48 AND pUL103 DURING LYTIC INFECTION

by

DANIEL A. ORTIZ

December 2015

Advisor: Dr. Philip E. Pellett

Major: Immunology and Microbiology

Degree: Doctor of Philosophy

Human cytomegalovirus (HCMV) is a large double-stranded DNA virus that causes severe disease in newborns and immunocompromised patients. During infection, HCMV is able to reconfigure the host cell machinery to establish a virus producing factory, termed the cytoplasmic virion assembly complex (cVAC). Generating drugs that affect cVAC development or function provides an alternative mode of action for HCMV antivirals that can essentially eliminate virion production. The objective of …

Infection Of Mammalian Hepatocytes By Francisella Tularensis Lvs, Pavel Vasioutovitch

Faculty Publications and Presentations

Francisella tularensis, the causative agent of Tularemia, is a facultative intracellular parasite. The goal of this project is to examine how F. tularensis infects and replicates within mammalian cells, and the impact of such an infection upon the signaling pathways within the host cell. The murine embryonic hepatocyte cell line TIB-73 serves as our model system for infection by F. tularensis live vaccine strain (LVS). The number of bacteria associated with hepatocytes was quantified via adhesion, invasion and intracellular replication assays. We have determined that a large multiplicity of infection (MOI) of F. tularensis LVS is required to infect …

Human Anatomy And Physiology Preparatory Course: Part 3 Of 4 (Interactive), Carlos Liachovitzky

Open Educational Resources

he overall purpose of these preparatory course set of learning objectives is to help students familiarize with some terms and some basic concepts they will find later in the Human Anatomy and Physiology I course.

These 40+ learning objectives to prepare for Human Anatomy and Physiology can be downloaded and played in a desktop, or laptop (windows exe file).

The entire course has four parts:

• Part 1
• Part 2
• Part 3
• Part 4

Each learning objective is followed by a set of multiple choice question similar to those found later in a Human Anatomy and Physiology course.

Radio-Frequency Plasma Polymerized Biodegradable Carrier For In Vivo Release Of Cis-Platinum, Sudhir Bhatt, Fatemeh Valamanesh, Jerome Pulpytel, Rea Lo Dico, Aliby Baiyukha, Iman Al-Dybiat, Marc Pocard, Farzaneh Arefi-Khonsari, Massoud Mirshahi

Bioelectrics Publications

A low pressure plasma process based on plasma deposition has been used to develop a drug delivery strategy. In this study, a drug delivery system based on different layers of plasma co-polymerized Poly ε-caprolactone-Polyethylene glycol (PCL-PEG) co-polymers was deposited on biocompatible substrates. Cis-platinum (118 μgm/cm²) was used as an anti-cancer drug and incorporated for local delivery of the chemotherapeutic agent. The co-polymer layers and their interaction with cancer cells were analyzed by scanning electron microscopy. Our study showed that the plasma-PCL-PEG coated cellophane membranes, in which the drug, was included did not modify the flexibility and appearance of …

The Impact Of Oxygen On Bacterial Enteric Pathogens, Nathan Wallace, Ashley N. Zani, Elizabeth A. Abrams, Yvonne Sun

Biology Faculty Publications

Bacterial enteric pathogens are responsible for a tremendous amount of foodborne illnesses every year through the consumption of contaminated food products. During their transit from contaminated food sources to the host gastrointestinal tract, these pathogens are exposed and must adapt to fluctuating oxygen levels to successfully colonize the host and cause diseases. However, the majority of enteric infection research has been conducted under aerobic conditions. To raise awareness of the importance in understanding the impact of oxygen, or lack of oxygen, on enteric pathogenesis, we describe in this review the metabolic and physiological responses of nine bacterial enteric pathogens exposed …

Comparative Proteomic Analysis Of Extracellular Vesicles From Prostate Cancer-Derived Cell Lines, Gloria Polanco

Open Access Theses & Dissertations

Prostate cancer (PCa) is the leading non-cutaneous malignancy and the second deadliest among American men. PCa mortality rates among African American men are much higher than any other ethnic group, and the same is true for men of African ancestry world-wide. There is also a lack of reliable diagnostic markers and effective treatment options. Extracellular vesicles (EVs) have been observed to play an important role in cancer processes such as promotion of tumor growth and metastasis. They are also a promising source of diagnostic markers. This study addresses these problems by studying the proteome of EVs derived from PCa cells …

Defining And Characterizing The Step In The Hiv-1 Viral Life Cycle Affected By Fullerene Derivatives, Zachary Sean Martinez

Open Access Theses & Dissertations

Fullerene derivatives, both C60 and C70, were synthesized and characterized to test their activity against HIV-1 infection. The derivatives tested have previously been reported to inhibit HIV-1 replication and/or in vitro activity of reverse transcriptase. It was found that viruses produced in the presence of low micromolar concentrations of most of these fullerene derivatives exhibit a reduction in viral infectivity greater than 99%. Quantification of virion-associated viral RNA and p24 indicates that RNA packaging and viral production were unremarkable in these viruses. These compounds did not affect infectivity of mature virions, indicating no effect on the early steps of the …

Elucidating The Molecular Mechanism Of Action By Which Rs1-208b Induces Apoptosis In Hematological Cancers, Ruben Israel Calderon

Open Access Theses & Dissertations

The 26S proteasome has been successfully targeted to treat multiple myeloma. However, the effect can sometimes be reversed as cells acquire resistance to chemotherapy. Recently, combining proteasome inhibitors with indirect proteasome inhibitors has been proposed in order to delay and even reverse resistance. Small molecules derived from 4-piperidinone that maintain an unsaturated β-carbonyl group are thought to interact with the thiol group of catalytically active cysteine residues in deubiquitinases that associate with the 26S proteasome, effectively inhibiting the proteasome.

RS1-208b was selected following a high-throughput screening of a small library of 4-piperidinone derived curcumin structural analogs due to its selectivity …

Human Anatomy And Physiology Preparatory Course: Part 1 Of 4 (Interactive), Carlos Liachovitzky

Open Educational Resources

The overall purpose of these preparatory course set of learning objectives is to help students familiarize with some terms and some basic concepts they will find later in the Human Anatomy and Physiology I course.

These 40+ learning objectives to prepare for Human Anatomy and Physiology can be downloaded and played in a desktop, or laptop (windows exe file).

The entire course has four parts:

• Part 1
• Part 2
• Part 3
• Part 4

Each learning objective is followed by a set of multiple choice question similar to those found later in a Human Anatomy and Physiology course.

The organization and …

Human Anatomy And Physiology Preparatory Course: Part 4 Of 4 (Interactive), Carlos Liachovitzky

Open Educational Resources

The overall purpose of these preparatory course set of learning objectives is to help students familiarize with some terms and some basic concepts they will find later in the Human Anatomy and Physiology I course.

These 40+ learning objectives to prepare for Human Anatomy and Physiology can be downloaded and played in a desktop, or laptop (windows exe file).

The entire course has four parts:

• Part 1
• Part 2
• Part 3
• Part 4

Each learning objective is followed by a set of multiple choice question similar to those found later in a Human Anatomy and Physiology course.

The Role Of E-Cadherin Force In The Maintenance Of Homeostasis In Epithelial Acini, Fnu Vani Narayanan

Theses and Dissertations

Numerous three-dimensional model systems have emerged for emulating the biochemical and physiological states of native tissue. Yet little is known about the effects of mechanical forces on cell behavior in the context of an organized tissue structure in three-dimensional cell-culture. Epithelial cells cultured in a three-dimensional environment comprised of extracellular matrix proteins form spheroids of polarized cells. Cellular responses to mechanical cues, generated from dynamic interactions with the extracellular matrix and neighboring cells, are known to influence cellular behavior to a great extent. Previous studies have shown that tumorigenic progression has been frequently linked to the down regulation of E-cadherin, …

Intrinsic And Extrinsic Regulation Of Pineal Melatonin Rhythms, Ye Li

Theses and Dissertations--Biology

Circadian rhythm is a biological rhythm with period of about 24 hours. Circadian rhythm is universal in phyla from bacteria to mammals and exists in different level from gene expression to behavior. Circadian system consists of three components: 1) a self-sustained oscillator; 2) an input pathway which can alter the phase of the oscillator; and 3) an output such as gene expression, enzyme activity, hormone production, heart rate, body temperature or locomotor activities. The way the oscillator regulates its outputs is complicated, in that on one hand usually the oscillator is not the only factor affecting the outputs, and on …

Hydrogen Peroxide Induced Loss Of Heterozygosity Correlates With Replicative Lifespan And Mitotic Asymmetry In Saccharomyces Cerevisiae, Emine Guven, Lindsay A. Parnell, Erin D. Jackson, Meighan Parker, Nilin Gupta, Jenny Rodrigues, Hong Qin

Scholarly Works

Cellular aging in Saccharomyces cerevisiae can lead to genomic instability and impaired mitotic asymmetry. To investigate the role of oxidative stress in cellular aging, we examined the effect of exogenous hydrogen peroxide on genomic instability and mitotic asymmetry in a collection of yeast strains with diverse backgrounds. We treated yeast cells with hydrogen peroxide and monitored the changes of viability and the frequencies of loss of heterozygosity (LOH) in response to hydrogen peroxide doses. The mid-transition points of viability and LOH were quantified using sigmoid mathematical functions. We found that the increase of hydrogen peroxide dependent genomic instability often occurs …

Impact Of Transcription Factors On Fibroblast Specific Gene Silencing In Somatic Cell Hybrids, Tania Ray

Masters Theses

As totipotent cells differentiate to specific cell types in multicellular organisms, certain sets of genes are turned off (gene silencing/gene extinction) while others are turned on (gene activation). Cell type specific gene silencing and gene activation is the basis of cell differentiation. Somatic cell hybrids which are produced by fusing different types of somatic cells from two different tissues have been used as a valuable resource to discern the phenomenon of lineage- specific gene extinction. Identification of regulatory factors that mediate gene extinction in hybrids are key to understanding regulatory mechanisms that govern cell differentiation in a multicellular organism. Transcription …

Loss Of Rap1gap: A Driver In The Progression From Dcis Of The Breast To Idc, Seema Shah

Wayne State University Dissertations

The purpose of this study is to determine the role of Rap1Gap in the progression of DCIS (ductal carcinoma in situ) to IDC (invasive ductal cancer). We employed an in-vitro three-dimensional (3D) overlay model that provides a physiologically relevant microenvironment to study mechanisms of malignant progression. Previous studies from this laboratory aimed to determine specific gene expression changes common to three models of DCIS- MCF10.DCIS, SUM 102 and SUM 225 in comparison to MCF10A cells, a model of non-transformed human mammary epithelium. The expression of 295 genes was found to be significantly altered, with 63 being increased in expression in …