Cell and Developmental Biology Commons^™

Structure-Function Analysis Of Nel, A Thrombospondin-1-Like Glycoprotein Involved In Neural Development And Functions, Masaru Nakamoto

Biology Faculty Publications

Nel (neural epidermal growth factor (EGF)-like molecule) is a multimeric, multimodular extracellular glycoprotein with heparin-binding activity and structural similarities to thrombospondin-1. Nel is predominantly expressed in the nervous system and has been implicated in neuronal proliferation and differentiation, retinal axon guidance, synaptic functions, and spatial learning. The Nel protein contains an N-terminal thrombospondin-1 (TSP-N) domain, five cysteine-rich domains, and six EGF-like domains. However, little is known about the functions of specific domains of the Nel protein. In this study, we have performed structure-function analysis of Nel, by using a series of expression constructs for different regions of the Nel protein. …

Effects Of Binge Alcohol Exposure On Canonical Wnt Signaling During Fracture Repair, Kristen Leigh Lauing

Dissertations

Excessive alcohol consumption is associated with increased fracture risk, delayed bone healing and fracture non-union. Binge alcohol consumption has occurred in 25-40% of all orthopaedic trauma cases. The canonical Wnt pathway, through tight regulation of stabilized beta-catenin, plays an essential and pivotal role in the formation of new bone and cartilage to initiate bone repair. We sought to determine the molecular mechanisms behind alcohol-related fracture complications by testing the hypothesis that binge alcohol exposure deregulates canonical Wnt signaling in the fracture callus, leading to impaired healing.

To test this, C57BL/6 or beta-catenin/TCF-reporter male mice were exposed to intraperitoneal alcohol or …

Triticum Mosaic Poacevirus Enlists P1 Rather Than Hc-Pro To Suppress Rna Silencing-Mediated Host Defense, Satyanarayana Tatineni, Feng Qu, Ruhui Li, Thomas Jack Morris, Roy French

Nebraska Center for Virology: Faculty Publications

Triticum mosaic virus (TriMV) is the type species of the newly established Poacevirus genus in the family Potyviridae. In this study, we demonstrate that in contrast to the helper component- proteinase (HC-Pro) of Potyvirus species, the P1 proteins of TriMV and Sugarcane steak mosaic poacevirus function in suppression of RNA silencing (SRS). TriMV P1 effectively suppressed silencing induced by single- or double-stranded RNAs (ss/ds RNAs), and disrupted the systemic spread of silencing signals at a step after silencing signal production. Interestingly, contrary to enhanced SRS activity of potyviral HC-Pro by co-expression with P1, the presence of TriMV HC-Pro reduced …

Identification Of Molecular Determinants From Moloney Leukemia Virus 10 Homolog (Mov10) Protein For Virion Packaging And Anti-Hiv-1 Activity, Aierken Abudu, Xiaojun Wang, Ying Dang, Tao Zhou, Shi-Hua Xiang, Yong -Hui Zheng

Nebraska Center for Virology: Faculty Publications

Background: MOV₁₀ inhibits HIV-1 replication after being packaged.

Results: A Gag binding plus all but one of seven helicase domains are required for MOV^₁₀ packaging. Nearly all residues are required for anti-HIV-1 activity. Conclusion: Gag binding is not sufficient for MOV₁₀ packaging, and multiple discontinuous domains regulate MOV₁₀ activity.

Significance: These findings uncover a new packaging mechanism and provide new insights into MOV₁₀ antiviral activity.

An Ims/Atp Assay For The Detection Of Mycobacterium Tuberculosis In Urine, Dawn M. Hunter, Daniel V. Lim

Molecular Biosciences Faculty Publications

Background. Although sputum smears are the gold standard for diagnosis of tuberculosis, sensitivity in HIV/TB coinfection cases is low, indicating a need for alternative methods. Urine is being increasingly evaluated. Materials and Methods. A novel method for detecting Mycobacterium tuberculosis (MTB) in synthetic urine using a combined IMS/ATP assay was evaluated. Preliminary work established standard ATP conditions and the sensitivity and specificity of the MTB antibody. Eighty-four blinded samples in four replicate assays were evaluated for the presence of MTB using labeled immunomagnetic beads for capture. Beads were separated, washed, and resuspended in broth and added to a …

Identification Of New Cell Size Control Genes In S. Cerevisiae, Huzefa Dungrawala, Hui Hua, Jill Wright, Lesley Abraham, Thivakorn Kasemsri, Anthony Mcdowell, Jessica Stilwell, Brandt L. Schneider

Molecular Biosciences Faculty Publications

Cell size homeostasis is a conserved attribute in many eukaryotic species involving a tight regulation between the processes of growth and proliferation. In budding yeast S. cerevisiae, growth to a “critical cell size” must be achieved before a cell can progress past START and commit to cell division. Numerous studies have shown that progression past START is actively regulated by cell size control genes, many of which have implications in cell cycle control and cancer. Two initial screens identified genes that strongly modulate cell size in yeast. Since a second generation yeast gene knockout collection has been generated, we screened …

The Two-Component Signal Transduction System Arlrs Regulates Staphylococcus Epidermidis Biofilm Formation In An Ica-Dependent Manner, Yang Wu, Jiaxue Wang, Tao Xu, Jingran Liu, Wenqi Yu, Qiang Lou, Tao Zhu, Nianan He, Haijing Ben, Jian Hu, Friedrich Götz, Di Qu

Molecular Biosciences Faculty Publications

Due to its ability to form biofilms on medical devices, Staphylococcus epidermidis has emerged as a major pathogen of nosocomial infections. In this study, we investigated the role of the two-component signal transduction system ArlRS in regulating S. epidermidis biofilm formation. An ArlRS-deficient mutant, WW06, was constructed using S. epidermidis strain 1457 as a parental strain. Although the growth curve of WW06 was similar to that of SE1457, the mutant strain was unable to form biofilms in vitro. In a rabbit subcutaneous infection model, sterile disks made of polymeric materials were implanted subcutaneously followed with inoculation of WW06 or SE1457. …

Wipi-1 Positive Autophagosome-Like Vesicles Entrap Pathogenic Staphylococcus Aureus For Lysosomal Degradation, Mario Mauthe, Wenqi Yu, Oleg Krut, Martin Krönke, Friedrich Götz, Horst Robenek, Tassula Proikas-Cezanne

Molecular Biosciences Faculty Publications

Invading pathogens provoke the autophagic machinery and, in a process termed xenophagy, the host cell survives because autophagy is employed as a safeguard for pathogens that escaped phagosomes. However, some pathogens can manipulate the autophagic pathway and replicate within the niche of generated autophagosome-like vesicles. By automated fluorescence-based high content analyses, we demonstrate that Staphylococcus aureus strains (USA300, HG001, SA113) stimulate autophagy and become entrapped in intracellular PtdIns(3)P-enriched vesicles that are decorated with human WIPI-1, an essential PtdIns(3)P effector of canonical autophagy and membrane protein of both phagophores and autophagosomes. Further, agr-positive S. aureus (USA300, HG001) strains were more efficiently …

Understanding Fracture, Allison Marsh

Section 3: Imaging the Fast Moving

No abstract provided.

Standardised Classification Of Pre-Release Development In Male-Brooding Pipefish, Seahorses, And Seadragons (Family Syngnathidae), Stefan Sommer, Camilla M. Whittington, Anthony B. Wilson

Publications and Research

Background: Members of the family Syngnathidae share a unique reproductive mode termed male pregnancy. Males carry eggs in specialised brooding structures for several weeks and release free-swimming offspring. Here we describe a systematic investigation of pre-release development in syngnathid fishes, reviewing available data for 17 species distributed across the family. This work is complemented by in-depth examinations of the straight-nosed pipefish Nerophis ophidion, the black-striped pipefish Syngnathus abaster, and the potbellied seahorse Hippocampus abdominalis.

Results: We propose a standardised classification of early syngnathid development that extends from the activation of the egg to the release of newborn. The classification consists …

One-Pot Regioselective Synthesis Of Tetrahydroindazolones And Evaluation Of Their Anti-Proliferative And Src Kinase Inhibitory Activities, V. Kameshwara Rao, Bhupender S. Chhikara, Rakesh Tiwari, Amir Nasrolahi Shirazi, Keykavous Parang, Anil Kumar

Pharmacy Faculty Articles and Research

A number of 2-substituted tetrahydroindazolones were synthesized by three-component condensation reaction of 1,3- diketones, substituted hydrazines, benzaldehydes, and Yb(OTf)3 as a catalyst in [bmim][BF4] ionic liquid using a simple, efficient, and economical one-pot method. The synthesized tetrahydroindazolones were evaluated for inhibition of cell proliferation of human colon carcinoma (HT-29), human ovarian adenocarcinoma (SK-OV-3), and c-Src kinase activity. 3,4-Dichlorophenyl tetrahydroindazolone derivative (15) inhibited the cell proliferation of HT-29 and SK-OV-3 cells by 62% and 58%, respectively. 2,3-Diphenylsubstituted tetrahydroindazolone derivatives, 19, 25, and 33, inhibited the cell proliferation of HT-29 cells by 6572% at a concentration of 50 μM. In general, the …

Enhanced Killing Effect Of Nanosecond Pulse Electric Fields On Panc1 And Jurkat Cell Lines In The Presence Of Tween 80, Gaurav Basu, Bhargava Subhash Kalluri, Ahmet Can Sabuncu, Christopher J. Osgood, Michael W. Stacey

Bioelectrics Publications

We investigated the effects of nanosecond pulse electric fields (nsPEFs) on Jurkat and PANC1 cells, which are human carcinoma cell lines, in the presence of Tween 80 (T80) at a concentration of 0.18% and demonstarted an enhanced killing effect. We used two biological assays to determine cell viability after exposing cells to nsPEFs in the presence of T80 and observed a significant increase in the killing effect of nsPEFs. We did not see a toxic effect of T80 when cells were exposed to surfactant alone. However, we saw a synergistic effect when cells exposed to T80 were combined with the …

Probing Nanoparticle Interactions In Cell Culture Media, Ahmet C. Sabuncu, Janna Grubbs, Shizhi Qian, Tarek M. Abdel-Fattah, Michael W. Stacey, Ali Beskok

Bioelectrics Publications

Nanoparticle research is often performed in vitro with little emphasis on the potential role of cell culture medium. In this study, gold nanoparticle interactions with cell culture medium and two cancer cell lines (human T-cell leukemia Jurkat and human pancreatic carcinoma PANC1) were investigated. Gold nanoparticles of 10, 25, 50, and 100 nm in diameter at fixed mass concentration were tested. Size distributions and zeta potentials of gold nanoparticles suspended in deionized (DI) water and Dulbecco's Modified Eagle's Media (DMEM) supplemented with fetal calf serum (FCS) were measured using dynamic light scattering (DLS) technique. In DI water, particle size distributions …

Inhibition Of Voltage-Gated Na+ Current By Nanosecond Pulsed Electric Field (Nspef) Is Not Mediated By Na+ Influx Or Ca²+ Signaling, Vasyl Nesin, Andrei G. Pakhomov

Bioelectrics Publications

In earlier studies, we found that permeabilization of mammalian cells with nsPEF was accompanied by prolonged inhibition of voltage-gated (VG) currents through the plasma membrane. This study explored if the inhibition of VG Na⁺ current (I_Na) resulted from (i) reduction of the transmembrane Na⁺ gradient due to its influx via nsPEF-opened pores, and/or (ii) downregulation of the VG channels by a Ca²⁺ -dependent mechanism. We found that a single 300?ns electric pulse at 1.65.3?kV/cm triggered sustained Na⁺ influx in exposed NG108 cells and in primary chromaffin cells, as detected by increased fluorescence of a …

Electric Field Exposure Triggers And Guides Formation Of Pseudopod-Like Blebs In U937 Monocytes, Mikhail A. Rassokhin, Andrei G. Pakhomov

Bioelectrics Publications

We describe a new phenomenon of anodotropic pseudopod-like blebbing in U937 cells stimulated by nanosecond pulsed electric field (nsPEF). In contrast to "regular," round-shaped blebs, which are often seen in response to cell damage, pseudopod-like blebs (PLBs) formed as longitudinal membrane protrusions toward anode. PLB length could exceed the cell diameter in 2 min of exposure to 60-ns, 10-kV/cm pulses delivered at 10-20 Hz. Both PLBs and round-shaped nsPEF-induced blebs could be efficiently inhibited by partial isosmotic replacement of bath NaCl for a larger solute (sucrose), thereby pointing to the colloid-osmotic water uptake as the principal driving force for bleb …

Oxidative Effects Of Nanosecond Pulsed Electric Field Exposure In Cells And Cell-Free Media, Olga N. Pakhomova, Vera A. Khorokhorina, Angela M. Bowman, Raminta Rodaitė-Riševičienė, Gintautas Saulis, Shu Xiao, Andrei G. Pakhomov

Bioelectrics Publications

Nanosecond pulsed electric field (nsPEF) is a novel modality for permeabilization of membranous structures and intracellular delivery of xenobiotics. We hypothesized that oxidative effects of nsPEF could be a separate primary mechanism responsible for bioeffects. ROS production in cultured cells and media exposed to 300-ns PEF (1–13 kV/cm) was assessed by oxidation of 2′, 7′-dichlorodihydrofluoresein (H₂DCF), dihidroethidium (DHE), or Amplex Red. When a suspension of H₂DCF-loaded cells was subjected to nsPEF, the yield of fluorescent 2′,7′dichlorofluorescein (DCF) increased proportionally to the pulse number and cell density. DCF emission increased with time after exposure in nsPEF-sensitive Jurkat …

The Development And Morphology Of Zebrafish After Embryonic Ethanol Exposure, Frances Lee

Honors Theses

Maternal consumption of alcohol may subject the fetus to fetal alcohol syndrome or fetal alcohol spectrum disorders (FAS or FASD). FAS/D is a public health problem, and affected children are defined by varying degrees of irreversible mental retardation, physical defects, behavioral issues, and vision problems from prenatal alcohol exposure (Riley et al. 2011). Recent studies on FAS have looked towards animal models, such as zebrafish, Danio rerio, that exhibit homologous physical and behavioral effects of alcohol (Bilotta et al. 2004). I exposed zebrafish embryos to low doses of ethanol (0.5% v/v or 1% v/v) in either chronic (at least 8 …

Desiccation Tolerance In Insect Cells, John Marcus Anderson

Masters Theses

In anhydrobiotic animals a reoccurring strategy to survive water stress involves the accumulation of low molecular weight solutes (e.g., trehalose) and highly hydrophilic macromolecules [e.g., late embryogenesis abundant (LEA) proteins]. We utilized two different water-stress models (spin-drying, and hyperosmotic dehydration) to investigate the impact of transgenic expression of a trehalose transporter [BT003466 (TRET)] and of a group 1 LEA protein [ACX81198 variant (LEA1)] on membrane integrity in cell lines from Spodoptera frugiperda (Sf-21) and Drosophila melanogaster (Kc167), respectively. Kc167 cells were challenged with sucrose or trehalose concentrations ranging from 0 to 200 mM supplemented in standard culture medium. Significant inhibition …

Melatonin Membrane Receptors In Peripheral Tissues: Distribution And Functions, Radomir M. Slominski, Russel J. Reiter, Natalia Schlabritz-Loutsevich, Rennolds S. Ostrom, Andrzej T. Slominski

Pharmacy Faculty Articles and Research

Many of melatonin’s actions are mediated through interaction with the G-protein coupled membrane bound melatonin receptors type 1 and type 2 (MT1 and MT2, respectively) or, indirectly with nuclear orphan receptors from the RORα/RZR family. Melatonin also binds to the quinone reductase II enzyme, previously defined the MT3 receptor. Melatonin receptors are widely distributed in the body; herein we summarize their expression and actions in non-neural tissues. Several controversies still exist regarding, for example, whether melatonin binds the RORα/RZR family. Studies of the peripheral distribution of melatonin receptors are important since they are attractive targets for immunomodulation, regulation of endocrine, …

Hyperosmotic Stress Enzyme Signaling Modulates Oct4, Nanog, And Rex1 Expression And Induces Prioritized Differentiation Of Murine Embryonic Stem Cells, Jill A. Slater

Wayne State University Dissertations

HYPEROSMOTIC STRESS ENZYME SIGNALING MODULATES OCT4, NANOG,

AND REX1 EXPRESSION AND INDUCES PRIORITIZED DIFFERENTIATION OF

MURINE EMBRYONIC STEM CELLS

by

JILL SLATER

MAY 2013

Advisor: Daniel Rappolee, Ph.D.

Major: Physiology

Degree: Doctor of Philosophy

Transcription factor expression and therefore lineage identity in the periimplantation

embryo and its stem cells may be influenced by extracellular stresses,

potentially affecting pregnancy outcome. Cellular stress forces cells to suppress some

normal activities (such as protein synthesis and cell proliferation) in order to repair

stress-damaged macromolecules and restore homeostasis. Therefore, any new

activities that embryonic cells initiate while concurrently funding the demands of the

stress …

Identification Of Trophic And Transcriptional Requirements For Human Embryonic Stem Cell Culture And Guided Neural Differentiation, Nicolas Edward Tokas

Legacy Theses & Dissertations (2009 - 2024)

Translational biomedical research holds potential for the development of novel therapeutics for the treatment of injury, illness or disability. To better understand intrinsic cellular and molecular biology, human embryonic stem cells (hESCs) can be used as a tool. hESCs are pluripotent with the abilities to be held in a self-renewal state or become induced to a desired germ layer then to be later directed through differentiation.

The Role Of An Rna Binding Protein Hnrnp K During Axon Development And Regeneration In Xenopus Laevis, Yuanyuan Liu

Legacy Theses & Dissertations (2009 - 2024)

Coordinated synthesis and assembly of the cytoskeletal network contribute significantly to morphological changes during axon outgrowth. Previous studies demonstrated that heterogeneous ribonucleoprotein K (hnRNP K), an RNA binding protein, binds to the 3'-untranslated regions of all neurofilament triplet subunits, the most abundant components of the axonal cytoskeleton. These findings raised the hypothesis that hnRNP K post-transcriptionally mediates the coordinated expression of axonal cytoskeletal components. In my thesis, I test this hypothesis during both axonal development and regeneration.

Dynamics Of Envelope Evolution In Clade C Shiv-Infected Pig-Tailed Macaques During Disease Progression Analyzed By Ultra-Deep Pyrosequencing, For Yue Tso, Damien C. Tully, Sandra Gonzalez, Christopher Quince, On Ho, Patricia Polacino, Ruth M. Ruprecht, Shiu-Lok Hu, Charles Wood

Nebraska Center for Virology: Faculty Publications

Understanding the evolution of the human immunodeficiency virus type 1 (HIV-1) envelope during disease progression can provide tremendous insights for vaccine development, and simian-human immunodeficiency virus (SHIV) infection of nonhuman primate provides an ideal platform for such studies. A newly developed clade C SHIV, SHIV-1157ipd3N4, which was able to infect rhesus macaques, closely resembled primary HIV-1 in transmission and pathogenesis, was used to infect several pig-tailed macaques. One of the infected animals subsequently progressed to AIDS, whereas one remained a nonprogressor. The viral envelope evolution in the infected animals during disease progression was analyzed by a bioinformatics approach using ultra-deep …

Pax6 Is Required For The Development Of The Lateral Procephalon In Tribolium, Qing Luan

Wayne State University Theses

In Tribolium, combinatorial knockdown of the Pax6 orthologs eyeless (ey) and twin of eyeless (toy) affects the peripheral visual system but also other areas of the dorsal larval head capsule. To elucidate the role of Pax6 genes during Tribolium embryonic head development in detail, we performed an extensive analysis of cuticle

elements, brain anatomy, embryonic head morphogenesis and developmental marker gene expression. Our results reveal that Pax6 is required for the development of a large contiguous area of the lateral anterior head, morphologically addressed as the embryonic head lobes, which encompass the neuroectodermal precursor tissues of the visual system, parts …

Genetics And Cancer, Sachin Puri

A with Honors Projects

Genes' effect in body and relationship with cancer. Role in cell cycle and angiogenesis.

Choreographing The Adenylyl Cyclase Signalosome: Sorting Out The Partners And The Steps, Rennolds S. Ostrom, Amy S. Bogard, Robert Gros

Pharmacy Faculty Articles and Research

Adenylyl cyclases are a ubiquitous family of enzymes and are critical regulators of metabolic and cardiovascular function. Multiple isoforms of the enzyme are expressed in a range of tissues. However, for many processes, the adenylyl cyclase isoforms have been thought of as essentially interchangeable, with their impact more dependent on their common actions to increase intracellular cyclic adenosine monophosphate content regardless of the isoform involved. It has long been appreciated that each subfamily of isoforms demonstrate a specific pattern of “upstream” regulation, i.e., specific patterns of ion dependence (e.g., calcium-dependence) and specific patterns of regulation by kinases (protein kinase A …

Kshv Infection Of Endothelial Cells Manipulates Cxcr7-Mediated Signaling: Implications For Kaposi’S Sarcoma Progression And Intervention, Jennifer Totonchy, Lisa Clepper, Janet Douglas, Liron Pantanowitz, Klaus Fruh, Ashlee V. Moses

Pharmacy Faculty Articles and Research

"CXCR7 was recently characterized as an alternative receptor for the chemokine CXCL12/SDF-1, previously thought to bind and signal exclusively through CXCR4.We recently identified CXCR7 as a key cellular factor in the endothelial cell (EC) dysfunction associated with KSHV infection. CXCL12 signaling is critically associated with tumor growth, angiogenesis and metastasis in several diverse tumors and is one of the most studied chemokine/chemokine receptor interactions in cancer systems. The tumorigenic activity of the CXCL12 signaling axis offers an attractive target for therapeutic intervention in multiple cancers including Kaposi’s Sarcoma (KS). However, most of the research to date was based on the …

Cytoplasmic Sequestration Of The Tumor Suppressor P53 By A Heat Shock Protein 70 Family Member, Mortalin, In Human Colorectal Adenocarcinoma Cell Lines, Erin E. Gestl, S. Anne Boettger

Biology Faculty Publications

No abstract provided.

An Exhibit Wordle, Allison Marsh

Handouts

No abstract provided.

A Crossword Puzzle For Kids, Allison Marsh

Handouts

No abstract provided.