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Genetic Analysis Reveals A Hierarchy Of Interactions Between Polycystin-Encoding Genes And Genes Controlling Cilia Function During Left-Right Determination, Daniel T. Grimes, Jennifer L. Keynton, Maria T. Buenavista, Xingjian Jin, Saloni H. Patel, Shinohara Kyosuke, Jennifer Vibert, Debbie J. Williams, Hiroshi Hamada, Rohana Hussain, Surya M. Nauli, Dominic P. Norris 2016 MRC Harwell

Genetic Analysis Reveals A Hierarchy Of Interactions Between Polycystin-Encoding Genes And Genes Controlling Cilia Function During Left-Right Determination, Daniel T. Grimes, Jennifer L. Keynton, Maria T. Buenavista, Xingjian Jin, Saloni H. Patel, Shinohara Kyosuke, Jennifer Vibert, Debbie J. Williams, Hiroshi Hamada, Rohana Hussain, Surya M. Nauli, Dominic P. Norris

Pharmacy Faculty Articles and Research

During mammalian development, left-right (L-R) asymmetry is established by a cilia-driven leftward fluid flow within a midline embryonic cavity called the node. This ‘nodal flow’ is detected by peripherally-located crown cells that each assemble a primary cilium which contain the putative Ca2+ channel PKD2. The interaction of flow and crown cell cilia promotes left side-specific expression of Nodal in the lateral plate mesoderm (LPM). Whilst the PKD2-interacting protein PKD1L1 has also been implicated in L-R patterning, the underlying mechanism by which flow is detected and the genetic relationship between Polycystin function and asymmetric gene expression remains unknown. Here, we ...


Investigating The Role Of Extracellular Matrix Proteins In Ovarian Folliculogenesis And Ovarian Cancer, Alexandra Carrier 2016 The University of Western Ontario

Investigating The Role Of Extracellular Matrix Proteins In Ovarian Folliculogenesis And Ovarian Cancer, Alexandra Carrier

Electronic Thesis and Dissertation Repository

The extracellular matrix (ECM) is a highly organized, dynamic structure that maintains tissue integrity and regulates biological processes involved in organ development and function. To explore the role of ECM proteins in ovarian physiology and pathology, my thesis characterizes ECM proteins aberrantly overexpressed in the Estrogen Receptor (ER)β-null (βERKO) mouse ovary and epithelial ovarian cancer (EOC). The ECM undergoes extensive physical changes, and influences numerous cell functions, throughout folliculogenesis. This study identifies a role for ERβ in ovarian development earlier than previously believed. Nidogen 2 and Collagen 11a1 are aberrantly overexpressed in βERKO ovaries as early as postnatal day ...


Epas1 Promotes Endothelial Cell Migration In An Irx3-Dependent Manner, Petrina K. Barnett 2016 Clark Atlanta University

Epas1 Promotes Endothelial Cell Migration In An Irx3-Dependent Manner, Petrina K. Barnett

Electronic Theses & Dissertations Collection for Atlanta University & Clark Atlanta University

Iroquois homeobox gene 3 (Irx3) is a transcription factor belonging to the Iroquois family of homeobox genes that is expressed in the embryonic organs of multiple organisms. Although much is known about Irx3 during embryogenesis, the cross talk of expression and function of Irx3 in peripheral vascular disease (PVD) remains to be investigated. Herein, we demonstrate that Endothelial PAS Domain 1 (EPAS1), which is involved in vasculogenesis and diseases associated with hypoxia, is an upstream regulator of Irx3, and this interaction contributes to the cellular movement of human microvascular endothelial cells (HMVECs). Genetic EPAS1 loss-of-function (LOF) studies in HMVECs resulted ...


Human Embryonic Stem Cell Research Vs. Alternative Stem Cell Research: Is There A Compromise?, Halle Herringshaw 2016 John Carroll University

Human Embryonic Stem Cell Research Vs. Alternative Stem Cell Research: Is There A Compromise?, Halle Herringshaw

Senior Honors Projects

No abstract provided.


Identification Of Potential Drug Targets In Cancer Signaling Pathways Using Stochastic Logical Models, Peican Zhu, Hamidreza Montazeri Aliabadi, Hasan Uludag, Jie Han 2016 Northwestern Polytechnical University

Identification Of Potential Drug Targets In Cancer Signaling Pathways Using Stochastic Logical Models, Peican Zhu, Hamidreza Montazeri Aliabadi, Hasan Uludag, Jie Han

Pharmacy Faculty Articles and Research

The investigation of vulnerable components in a signaling pathway can contribute to development of drug therapy addressing aberrations in that pathway. Here, an original signaling pathway is derived from the published literature on breast cancer models. New stochastic logical models are then developed to analyze the vulnerability of the components in multiple signalling sub-pathways involved in this signaling cascade. The computational results are consistent with the experimental results, where the selected proteins were silenced using specific siRNAs and the viability of the cells were analyzed 72 hours after silencing. The genes elF4E and NFkB are found to have nearly no ...


Improving The Efficiency Of Crispr-Cas9: The Expression & Purification Of Cas9, Elaine Shults 2016 Student

Improving The Efficiency Of Crispr-Cas9: The Expression & Purification Of Cas9, Elaine Shults

Undergraduate Honors Theses

p53 is a transcription factor that plays a significant role in cell cycle regulation, senescence, and apoptosis. The p53 protein has been widely studied for its link to human cancer, and is estimated to be mutated in ~50% of all human cancers. This project focused on generating a p53-null A549 cell line using the genome editing technique CRISPR (Clustered Regularly-Interspaced Short Palindromic Repeats)-Cas9. CRISPR-Cas9 was employed to generate a simultaneous knock-in/knockout of mCherry and WTp53, respectively. The mCherry gene was inserted at the WTp53 endogenous translational start site (TSS), thereby rendering WTp53 inactive and mCherry “active” whenever WTp53 ...


Infection Of Mammalian Hepatocytes By Francisella Tularensis Lvs, Pavel Vasioutovitch 2016 Liberty University

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 the ...


Three-Dimensional Confocal Microscopy Indentation Method For Hydrogel Elasticity Measurement, Donghee Lee, Md Mahmudur Rahman, You Zhou, Sangjin Ryu 2015 University of Nebraska-Lincoln

Three-Dimensional Confocal Microscopy Indentation Method For Hydrogel Elasticity Measurement, Donghee Lee, Md Mahmudur Rahman, You Zhou, Sangjin Ryu

Md Mahmudur Rahman

No abstract provided.


Detection Of Ubiquitination On Syk And Documenting Syk Stability, Izabela Mazur, Wen Horng Wang, Robert J. Geahlen 2015 Purdue University

Detection Of Ubiquitination On Syk And Documenting Syk Stability, Izabela Mazur, Wen Horng Wang, Robert J. Geahlen

The Summer Undergraduate Research Fellowship (SURF) Symposium

Post-translational modifications regulate the activities of proteins important to numerous diseases. Spleen Tyrosine Kinase (Syk) is particularly interesting to researchers because it modifies many targets and plays multiple roles in regulating cells in our bodies and its abnormal modifications may contribute to cancer, Alzheimer’s disease and allergies. In an attempt to study these modifications of Syk, we first looked at detecting ubiquitination on Syk protein. Ubiquitin, a small 8 kDa molecule, attaches to lysine residues on protein. The attachment of ubiquitin to Syk may cause Syk to either propagate signals onwards to activate other proteins or signal it to ...


The Wings Of Bombyx Mori Develop From Larval Discs Exhibiting An Early Differentiated State: A Preliminary Report, Madhuri Kango-Singh, Amit Singh, K. P. Gopinathan 2015 University of Dayton

The Wings Of Bombyx Mori Develop From Larval Discs Exhibiting An Early Differentiated State: A Preliminary Report, Madhuri Kango-Singh, Amit Singh, K. P. Gopinathan

Madhuri Kango-Singh

Lepidopteran insects present a complex organization of appendages which develop by various mechanisms. In the mulberry silkworm, Bombyx mori, a pair of meso- and meta-thoracic discs located on either side in the larvae gives rise to the corresponding fore- and hind-wings of the adult. These discs do not experience massive cell rearrangements during metamorphosis and display the adult wing vein pattern. We have analysed wing development in B. mori by two approaches, viz., expression of patterning genes in larval wing discs, and regulatory capacities of larval discs following explantation or perturbation. Expression of Nubbin is seen all over the presumptive ...


The Wings Of Bombyx Mori Develop From Larval Discs Exhibiting An Early Differentiated State: A Preliminary Report, Madhuri Kango-Singh, Amit Singh, K. Gopinathan 2015 University of Dayton

The Wings Of Bombyx Mori Develop From Larval Discs Exhibiting An Early Differentiated State: A Preliminary Report, Madhuri Kango-Singh, Amit Singh, K. Gopinathan

Amit Singh

Lepidopteran insects present a complex organization of appendages which develop by various mechanisms. In the mulberry silkworm, Bombyx mori, a pair of meso- and meta-thoracic discs located on either side in the larvae gives rise to the corresponding fore- and hind-wings of the adult. These discs do not experience massive cell rearrangements during metamorphosis and display the adult wing vein pattern. We have analysed wing development in B. mori by two approaches, viz., expression of patterning genes in larval wing discs, and regulatory capacities of larval discs following explantation or perturbation. Expression of Nubbin is seen all over the presumptive ...


Cell Viability Assessment Using The Alamar Blue Assay: A Comparison Of 2d And 3d Cell Culture Models, Franck Bonnier, Mark Keating, Tomasz Wrobel, Katarsyna Majzner, Malgorzata Baranska, Amaya Garcia, Alfonso Blanco, Hugh Byrne 2015 Dublin Institute of Technology

Cell Viability Assessment Using The Alamar Blue Assay: A Comparison Of 2d And 3d Cell Culture Models, Franck Bonnier, Mark Keating, Tomasz Wrobel, Katarsyna Majzner, Malgorzata Baranska, Amaya Garcia, Alfonso Blanco, Hugh Byrne

Mark Keating

Comparisons of 2D and 3D cell culture models in literature have indicated differences in cellular morphology and metabolism, commonly attributed the better representation of in vivo conditions of the latter cell culture environment. Thus, interest in the use of 3D collagen gels for in vitro analysis has been growing. Although comparative studies to date have indicated an enhanced resistance of cells on collagen matrices against different toxicants, in the present study it is demonstrated that non-adapted protocols can lead to misinterpretation of results obtained from classical colorometric dye-based cytotoxic assays. Using the well established Alamar Blue assay, the study demonstrates ...


Epigenetic Silencing Of Id4 In Prostate Cancer: Mechanistic Insight, Swathi Chinaranagari 2015 Clark Atlanta University

Epigenetic Silencing Of Id4 In Prostate Cancer: Mechanistic Insight, Swathi Chinaranagari

Electronic Theses & Dissertations Collection for Atlanta University & Clark Atlanta University

Inhibitor of DNA binding/differentiation protein 4 (ID4) is a dominant negative regulator of basic helix loop helix (bHLH) family of transcription factors. ID4 shares the homology of HLH domain with other ID proteins (ID1, ID2, and ID3) and lack the basic DNA binding region. Evidence suggested that unlike ID1, ID2 and ID3, ID4 acts as a tumor suppressor in prostate cancer by attenuating cell proliferation and promoting apoptosis. Consistent with these observations ID4 is epigenetically silenced in DU145 prostate cancer cell line. In this study we investigated whether ID4 is also epigenetically silenced in prostate cancer. We also examined ...


Characterization And Transplantation Of Felid Spermatogonial Stem Cells, Robin H. Powell 2015 University of New Orleans

Characterization And Transplantation Of Felid Spermatogonial Stem Cells, Robin H. Powell

University of New Orleans Theses and Dissertations

Spermatogonial stem cells (SSC) self-renew and differentiate into spermatozoa throughout the life of the male. SSC transplantation is a potential method for the propagation of genetically important males. These cells have been isolated in different mammalian species using specific cell surface markers, but not in felines.

The goal of this study was to explore a relevant strategy for conservation of endangered felids by characterizing domestic cat (Felis catus) SSCs and assessing their ability for self-renewal after transplantation. Firstly, SSC and pluripotent surface markers, identified in non-feline species, were tested for expression in mixed germ cells from adults by immunocytochemistry and ...


Manipulation Of The Microbiome And Its Impact On Functional Recovery Following Ischemic Stroke, Michal Jandzinski 2015 University of Connecticut

Manipulation Of The Microbiome And Its Impact On Functional Recovery Following Ischemic Stroke, Michal Jandzinski

Honors Scholar Theses

Each year, nearly 800,000 individuals residing in the United States will have a stroke. Of these, about 130,000 cases will prove fatal while many of the survivors will be forced to live with disability for the remainder of their lives. Out of all strokes over 87% are ischemic strokes. The widespread incidence of this debilitating condition costs the United States an estimated $36.5 billion dollars every single year. Despite this, clinicians are armed with very little to combat the disease. Recent research developments have brought about the rise in awareness about the importance of the microbiome, the ...


Decellularized Basement Matrix Membranes And Their Ability To Support Engraftment And Proliferation Of Human Dermal Fibroblasts, Lauren A. Miller 2015 Kent State University - Kent Campus

Decellularized Basement Matrix Membranes And Their Ability To Support Engraftment And Proliferation Of Human Dermal Fibroblasts, Lauren A. Miller

Undergraduate Research Symposium

Decellularized extracellular matrix components isolated from animal sources have shown much clinical promise in the treatment of several wound care indications including a variety of dermal ulcerations (diabetic, pressure, venous, e.g.), full-thickness trauma wounds affecting the skin and skeletal muscle, and post-operative surgical wounds. The purpose of this study was to evaluate the biocompatibility of three sources of decellularized matrix wound care products using a novel tissue engineered dermal cellular proliferation assay. Deceullarized basement matrices derived from porcine small intestinal submucosa (pSIS), porcine peritoneal membrane (pPM), and ovine forestomach (oFS), respectively, were tested for their ability to support the ...


Cell Viability Assessment Using The Alamar Blue Assay: A Comparison Of 2d And 3d Cell Culture Models, Franck Bonnier, Mark Keating, Tomasz P. Wrobel, Katarsyna Majzner, Malgorzata Baranska, Amaya Garcia, Alfonso Blanco, Hugh Byrne 2015 Dublin Institute of Technology

Cell Viability Assessment Using The Alamar Blue Assay: A Comparison Of 2d And 3d Cell Culture Models, Franck Bonnier, Mark Keating, Tomasz P. Wrobel, Katarsyna Majzner, Malgorzata Baranska, Amaya Garcia, Alfonso Blanco, Hugh Byrne

Articles

Comparisons of 2D and 3D cell culture models in literature have indicated differences in cellular morphology and metabolism, commonly attributed the better representation of in vivo conditions of the latter cell culture environment. Thus, interest in the use of 3D collagen gels for in vitro analysis has been growing. Although comparative studies to date have indicated an enhanced resistance of cells on collagen matrices against different toxicants, in the present study it is demonstrated that non-adapted protocols can lead to misinterpretation of results obtained from classical colorometric dye-based cytotoxic assays. Using the well established Alamar Blue assay, the ...


Clique Topology Reveals Intrinsic Geometric Structure In Neural Correlations, Chad Giusti, Eva Pastalkova, Carina Curto, Vladimir Itskov 2015 University of Pennsylvania

Clique Topology Reveals Intrinsic Geometric Structure In Neural Correlations, Chad Giusti, Eva Pastalkova, Carina Curto, Vladimir Itskov

Faculty Publications, Department of Mathematics

Detecting meaningful structure in neural activity and connectivity data is challenging in the presence of hidden nonlinearities, where traditional eigenvalue-based methods may be misleading. We introduce a novel approach to matrix analysis, called clique topology, that extracts features of the data invariant under nonlinear monotone transformations. These features can be used to detect both random and geometric structure, and depend only on the relative ordering of matrix entries. We then analyzed the activity of pyramidal neurons in rat hippocampus, recorded while the animal was exploring a 2D environment, and confirmed that our method is able to detect geometric organization using ...


Detecting Cells In Time Varying Intensity Images In Confocal Microscopy For Gene Expression Studies In Living Cells, Debasis Mitra, Rostyslav Boutchko, Judhajeet Ray, Marit Nilsen-Hamilton 2015 Florida Institute of Technology

Detecting Cells In Time Varying Intensity Images In Confocal Microscopy For Gene Expression Studies In Living Cells, Debasis Mitra, Rostyslav Boutchko, Judhajeet Ray, Marit Nilsen-Hamilton

Biochemistry, Biophysics, and Molecular Biology Presentations and Posters

In this work we present a time-lapsed confocal microscopy image analysis technique for an automated gene expression study of multiple single living cells. Fluorescence Resonance Energy Transfer (FRET) is a technology by which molecule-to-molecule interactions are visualized. We analyzed a dynamic series of ~102 images obtained using confocal microscopy of fluorescence in yeast cells containing RNA reporters that give a FRET signal when the gene promoter is activated. For each time frame, separate images are available for three spectral channels and the integrated intensity snapshot of the system. A large number of time-lapsed frames must be analyzed to identify ...


Detection And Tracking Of T Cells In Time-Lapse Imaging, Cody Arbuckle, Milton L. Greenberg, Erik J. Linstead 2015 Chapman University

Detection And Tracking Of T Cells In Time-Lapse Imaging, Cody Arbuckle, Milton L. Greenberg, Erik J. Linstead

Mathematics, Physics, and Computer Science Faculty Articles and Research

The effective classification and tracking of cells obtained from modern staining techniques has significant limitations due to the necessity of having to train and utilize a human expert in the field who must manually identify each cell in each slide. Often times these slides are filled with noise cells that are not of particular interest to the researcher. The use of computational methods has the ability to effectively and efficiently enhance image quality, as well as identify and track target cell types over large data sets. Here we present a computational approach to the in vitro tracking of T cells ...


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