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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 J. 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 J. 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 ...


The Leishmania Years At Unl (Or, My Life As A Cell Biologist, 1966-1981), John J. Janovy Jr. 2015 University of Nebraska-Lincoln

The Leishmania Years At Unl (Or, My Life As A Cell Biologist, 1966-1981), John J. Janovy Jr.

Faculty Publications from the Harold W. Manter Laboratory of Parasitology

Slides for a talk during which Professor Janovy discussed the methods he used in researching Leishmania during the years 1966-1981. Includes lists of references.


Evaluation Of The Developmental Toxicity Of Lead In The Danio Rerio Body, Nicole M. Roy, Sarah DeWolf, Bruno Carneiro 2015 Sacred Heart University

Evaluation Of The Developmental Toxicity Of Lead In The Danio Rerio Body, Nicole M. Roy, Sarah Dewolf, Bruno Carneiro

Biology Faculty Publications

Lead has been utilized throughout history and is widely distributed and mobilized globally. Although lead in the environment has been somewhat mitigated, the nature of lead and its extensive uses in the past prohibit it from being completely absent from our environment and exposure to lead is still a public health concern. Most studies regarding lead toxicity have focused on the brain. However, little is found in the literature on the effects of lead in other tissues. Here, we utilize the zebrafish model system to investigate effects of lead exposure during early developmental time windows at 24, 48 and 72 ...


Mcnamara 201412 Nih Scap Innocentive Challenge Solution - T-Bow Rainbow T-Cells And Tumor Cells Spatial Multiplexing Gene Expression Reporter System – Plus Supplement Plus Posters - 20141228s, George McNamara 2014 M.D. Anderson Cancer Center

Mcnamara 201412 Nih Scap Innocentive Challenge Solution - T-Bow Rainbow T-Cells And Tumor Cells Spatial Multiplexing Gene Expression Reporter System – Plus Supplement Plus Posters - 20141228s, George Mcnamara

George McNamara

McNamara 201412 NIH SCAP InnoCentive Challenge Solution - T-Bow Rainbow T-cells and Tumor Cells Spatial Multiplexing Gene Expression Reporter System – plus supplement plus posters - 20141228S

The download is my/Cooper lab solution (submission) to the 2014 NIH Single Cell Analysis Program (SCAP) InnoCentive Challenge, "Follow That Cell". I submitted the Solution on 20141215Mon (with 20 minutes to spare). The Challenge web page is

https://www.innocentive.com/ar/challenge/9933618

Our Solution is to revolutionize live cell transcriptional reporter assays. Specifically, we propose to multiplex five promoter response elements to drive expression of different color fluorescent proteins that would bind to ...


Engineered Plga Nanoparticles For Delivery Of Sirna In Mcf-7 Breast Cancer Cells, Sydney Pong, Samit Shah, Vivek Gupta 2014 Chapman University

Engineered Plga Nanoparticles For Delivery Of Sirna In Mcf-7 Breast Cancer Cells, Sydney Pong, Samit Shah, Vivek Gupta

Undergraduate Student Research Day Abstracts and Posters

Small interfering RNAs have been an emerging medical treatment for molecular based diseases as they are capable of gene-specific knockdown. Appropriate and efficient delivery remains one of the biggest challenges in the development of siRNA as an anti-cancer treatment. Nanoparticles containing siRNA were characterized and the efficacy of various peptides in the transfection of the nanoparticles were tested. A gene silencing assay was developed in order to determine the effect of siRNA therapeutics on gene functionality in breast cancer cells.


Dancing Through Life: Allosteric Transitions And Structural Analysis Of Hsp70 And Hsp110 Chaperone Proteins, Gabrielle Stetz, Gennady M. Verkhivker 2014 Chapman University

Dancing Through Life: Allosteric Transitions And Structural Analysis Of Hsp70 And Hsp110 Chaperone Proteins, Gabrielle Stetz, Gennady M. Verkhivker

Undergraduate Student Research Day Abstracts and Posters

The molecular chaperone protein Hsp70 is centrally involved in cellular homeostasis by assisting in the folding and degradation of protein substrates. Hsp70 is joined by co-chaperones, such as Hsp110, which contribute to specialized tasks of the Hsp70 complex. Imbalances of this heat shock protein system are believed to be involved with the deregulation of cancer pathways and other human diseases. Better understanding of how these heat shock proteins work at the molecular level, which has been investigated using molecular docking tools, will give more clues about biological function. Simulating the formation and function of Hsp70 based chaperone complexes could provide ...


Prostate Field Cancerization -- Thinking Outside The Tumor, Dor Shoshan, Marco Bisoffi 2014 Chapman University

Prostate Field Cancerization -- Thinking Outside The Tumor, Dor Shoshan, Marco Bisoffi

Undergraduate Student Research Day Abstracts and Posters

Prostate field cancerization (or field effect) is characterized by the presence of molecular alterations in histologically normal tissues adjacent to adenocarcinomas. Accordingly, our research indicates deregulated expression of several proteins that define this type of molecular pathology. The scope of the present study was to determine the expression of the key transcription factor and potential marker of field cancerization early growth response 1 (EGR-1) in human prostate tissues derived from prostatectomies and biopsy cores.

EGR-1 was detected by immunofluorescence using a polyclonal anti-human EGR-1 and Alexa Fluor 488-conjugated secondary antibodies. EGR-1 expression was quantitated by determining the pixel count per ...


Toxicity And Cosmesis Outcomes For Single Fraction Intra-Operative Electron Radiotherapy (Ioert) For Breast Cancer, Monica Hanna, Robert Ash, Wesley Babaran, Michele M. Carpenter, Afshin Forouzannia, Jay K. Harness, Brian Kaltenecker, Snehith Maddula, Venita Williams, Lawrence Wagman 2014 Chapman University

Toxicity And Cosmesis Outcomes For Single Fraction Intra-Operative Electron Radiotherapy (Ioert) For Breast Cancer, Monica Hanna, Robert Ash, Wesley Babaran, Michele M. Carpenter, Afshin Forouzannia, Jay K. Harness, Brian Kaltenecker, Snehith Maddula, Venita Williams, Lawrence Wagman

Undergraduate Student Research Day Abstracts and Posters

Background: Adjuvant radiation therapy is proven to reduce local recurrence in patients with early stage breast cancer. To reduce toxicity, improve geographic accuracy, and reduce treatment time, IOERT can be utilized as an alternative to external beam radiation therapy. The study’s objective was to determine the short term toxicity and cosmesis profile of single fraction IOERT given as definitive treatment in a community setting. Materials and Methods: From Mar 2012 to Jul 2014, 84 patients (3 bilateral), ages 45-91 y.o. with stage 0-II were treated with IOERT (Mobetron, IntraOp Medical, Sunnyvale, CA). A single 21 Gy fraction was ...


Molecular Insights Into Prostate Field Cancerization: Telomere Length, Egr-­‐1 Expression, And Regulation Of Mic-­‐1, Pdgf-­‐A, And Fas, Emily Frisch, Kristin Gabriel, Marco Bisoffi 2014 Chapman University

Molecular Insights Into Prostate Field Cancerization: Telomere Length, Egr-­‐1 Expression, And Regulation Of Mic-­‐1, Pdgf-­‐A, And Fas, Emily Frisch, Kristin Gabriel, Marco Bisoffi

Undergraduate Student Research Day Abstracts and Posters

The diagnosis of prostate cancer (adenocarcinoma) relies on screening for elevated prostate-specific antigen (PSA) in blood samples and on digital rectal examination (DRE). With high PSA levels and/or abnormal DRE, physicians recommend a biopsy, which often misses the location of the adenocarcinoma and results in false negatives. Previous studies have shown expression of the key transcription factor early growth response 1 (EGR-1), the pro-survival factor macrophage inhibitor cytokine 1 (MIC-1), and the growth stimulatory platelet derived growth factor A (PDGF-A) to be up-regulated in histologically normal tissues 1 centimeter adjacent to prostate adenocarcinomas. We hypothesize that tumors emerge from ...


An Activity Aimed At Improving Student Explanations Of Biological Mechanisms, Caleb M. Trujillo, Trevor R. Anderson, Nancy J. Pelaez 2014 Purdue University

An Activity Aimed At Improving Student Explanations Of Biological Mechanisms, Caleb M. Trujillo, Trevor R. Anderson, Nancy J. Pelaez

PIBERG Instructional Innovation Materials

This document is intended for use by instructors and their students. The activity contains steps to introduce students to the MACH model involving analyzing and discussing explanations about biological mechanisms. Initially, students read modified articles about biological mechanisms during class, although instructors may prefer to assign readings outside of class before the activity. During the activity, students are required to analyze the readings for evidence of research methods, analogies, context, and mechanisms. In so doing, students learn how to integrate the information pertaining to each of the MACH model components into a coherent explanation about their biological mechanism. After performing ...


A Tetrahedral Version Of The Mach Model For Explaining Biological Mechanisms, Caleb M. Trujillo, Trevor R. Anderson, Nancy J. Pelaez 2014 Purdue University

A Tetrahedral Version Of The Mach Model For Explaining Biological Mechanisms, Caleb M. Trujillo, Trevor R. Anderson, Nancy J. Pelaez

PIBERG Instructional Innovation Materials

This document is intended for both instructors and students. Modified from the original MACH model this version, once cut and folded, creates a tetrahedral model that can conveniently be used as a teaching and learning tool to inform and guide students on how to write expert quality explanations of biological mechanisms. Each vertex of the tetrahedron represents a component of the model namely, Methods, Analogy, Context, and How. For a coherent and complete explanation about molecular mechanisms, it is important to integrate information pertaining to all four components of the model. The tetrahedral MACH model has been tested in both ...


Lkb1 Deficient Non-Small Cell Lung Cancer Cells Are Vulnerable To Energy Stress Induced By Atp Depletion, Chao Yang 2014 UT GSBS

Lkb1 Deficient Non-Small Cell Lung Cancer Cells Are Vulnerable To Energy Stress Induced By Atp Depletion, Chao Yang

UT GSBS Dissertations and Theses (Open Access)

Lung cancer is the second most frequent cancer in United States, which represents about 13.5% of new cancer cases every year. It accounts for about 27.2% of all cancer related deaths, which is more than the sum of deaths caused by prancretic, breast and colorectal. On average, only about 16% of lung cancer patients survive beyond 5 years. LKB1 is the third most mutated gene in lung cancer. It has been shown that LKB1 is mutated in at least 15% to 30% of NSCLC. Tumor with LKB1 mutation is associated with poor differentiation, high metastasis and worse response ...


The Drosophila T-Box Transcription Factor Midline Functions Within The Insulin/Akt And C-Jun-N-Terminal Kinase Signaling Pathways To Regulate Interomatidial Bristle Formation And Cell Survival, Qichuan Chen 2014 University of Southern Mississippi

The Drosophila T-Box Transcription Factor Midline Functions Within The Insulin/Akt And C-Jun-N-Terminal Kinase Signaling Pathways To Regulate Interomatidial Bristle Formation And Cell Survival, Qichuan Chen

Master's Theses

From a genetic and allelic modifier screen, we report that the Drosophila melanogaster T-box transcription factor midline (mid), a homolog to the human TBX20 gene, interacts with dFOXO within the insulin receptor (InR) and the c-Jun-N-terminal kinase (JNK) signaling pathways to regulate interommatidial bristle (IOB) formation. Previous studies have identified mid’s role in cell fate specification of sensory organ precursor cells in conjunction with the Notch-Delta signaling pathway (Das et al., 2013). The Notch, InR, and JNK signaling pathways regulate dFOXO activity under conditions of stress. Thus, we determined the effects of oxidative stress and metabolic stress by exposing ...


Structure-Based Network Analysis Of Activation Mechanisms In The Erbb Family Of Receptor Tyrosine Kinases: The Regulatory Spine Residues Are Global Mediators Of Structural Stability And Allosteric Interactions, Kevin A. James, Gennady M. Verkhivker 2014 Chapman University

Structure-Based Network Analysis Of Activation Mechanisms In The Erbb Family Of Receptor Tyrosine Kinases: The Regulatory Spine Residues Are Global Mediators Of Structural Stability And Allosteric Interactions, Kevin A. James, Gennady M. Verkhivker

Mathematics, Physics, and Computer Science Faculty Articles and Research

The ErbB protein tyrosine kinases are among the most important cell signaling families and mutation-induced modulation of their activity is associated with diverse functions in biological networks and human disease. We have combined molecular dynamics simulations of the ErbB kinases with the protein structure network modeling to characterize the reorganization of the residue interaction networks during conformational equilibrium changes in the normal and oncogenic forms. Structural stability and network analyses have identified local communities integrated around high centrality sites that correspond to the regulatory spine residues. This analysis has provided a quantitative insight to the mechanism of mutation-induced ‘‘superacceptor’’ activity ...


Gadph Single Molecule Mrna Fish 20141104tue, George McNamara 2014 M.D. Anderson Cancer Center

Gadph Single Molecule Mrna Fish 20141104tue, George Mcnamara

George McNamara

GADPH single molecule mRNA FISH 20141104Tue

Raw data on NVidia TITAN GPU card deconvolution using original Bruce and Butte 2013 Optics Express software running inside Fiji ImageJ.

B&B GPU Deconvolution Reference:

http://www.opticsinfobase.org/oe/fulltext.cfm?uri=oe-21-4-4766&id=249375

Acquired at 63x1x using leica plan apo 1.4 NA objective lens on Cooper lab Leica DMI6000 microscope, SOLA (initial release) LED lamp using only "green" LED, 554x Semrock exciter filter (in ASI ex wheel), Leica N3 (Cy3) filter cube, center 600 nm emission wavelength.

Hamamatsu FLASH4.0 sCMOS, 100 ms exposure time so 6.4 seconds ...


Majority Of Cells Lining The Walls Of The 3rd Ventricle In The Adult Rat Brain Are Not Neural Progenitor Cells, C. Hasken, S. Vermilyea, M. Hendrickson, R. Kalil 2014 DePauw University

Majority Of Cells Lining The Walls Of The 3rd Ventricle In The Adult Rat Brain Are Not Neural Progenitor Cells, C. Hasken, S. Vermilyea, M. Hendrickson, R. Kalil

Science Research Fellows Posters

No abstract provided.


Identifying Diagnostic Dna Methylation Profiles For Facioscapulohumeral Muscular Dystrophy In Blood And Saliva Using Bisulfite Sequencing, Takako I. Jones, Chi Yan, Peter Sapp, Diane McKenna-Yasek, Peter B. Kang, Colin Quinn, Johnny S. Salameh, Oliver D. King, Peter L. Jones 2014 University of Massachusetts Medical School

Identifying Diagnostic Dna Methylation Profiles For Facioscapulohumeral Muscular Dystrophy In Blood And Saliva Using Bisulfite Sequencing, Takako I. Jones, Chi Yan, Peter Sapp, Diane Mckenna-Yasek, Peter B. Kang, Colin Quinn, Johnny S. Salameh, Oliver D. King, Peter L. Jones

Peter Jones Lab

BACKGROUND: Facioscapulohumeral muscular dystrophy (FSHD) is linked to chromatin relaxation due to epigenetic changes at the 4q35 D4Z4 macrosatellite array. Molecular diagnostic criteria for FSHD are complex and involve analysis of high molecular weight (HMW) genomic DNA isolated from lymphocytes, followed by multiple restriction digestions, pulse-field gel electrophoresis (PFGE), and Southern blotting. A subject is genetically diagnosed as FSHD1 if one of the 4q alleles shows a contraction in the D4Z4 array to below 11 repeats, while maintaining at least 1 repeat, and the contraction is in cis with a disease-permissive A-type subtelomere. FSHD2 is contraction-independent and cannot be diagnosed ...


Tattletales And T-Bow Update 20141018, George McNamara 2014 M.D. Anderson Cancer Center

Tattletales And T-Bow Update 20141018, George Mcnamara

George McNamara

Tattletales and T-Bow Update 20141018

http://works.bepress.com/gmcnamara/63

Tattletales is my concept to multiplex fluorescent protein biosensors ("Tattletales", which is also the overall concept name) and multicolor FP reporters ("T-Bow" for Rainbow T-cells and tumor cells) to study the physiology of live cells.

These are based on:

1. LacI-GFP :: LacO operator synthetic tandem repeat array (Robinett et al 1996 JCB, Fig 4A, http://jcb.rupress.org/content/135/6/1685.full.pdf )

2. the plethora of fluorescent protein biosensors - see page 1 of this "63" download site.

3. the many colors, and combinations of colors (CY11.5 ...


Ligand-Directed Signaling And Trafficking At The Nociceptin Opioid Peptide Receptor, Gina E. Migneco 2014 Washington University in St. Louis

Ligand-Directed Signaling And Trafficking At The Nociceptin Opioid Peptide Receptor, Gina E. Migneco

Undergraduate Research Symposium Posters & Abstracts

Traditional opioid receptors such as the μ and κ opioid receptors (MOR and

KOR) are well studied and are found to be common targets for pain and

stress therapies. The nociceptin opioid receptor (NOPR/ORL1) is a most

recently discovered opioid receptor whose complete function is still relatively

unknown, however is of great interest to researchers due to its analgesic

and antidepressant properties. NOPR activation due to ligand binding

triggers a variety of downstream effects, including G-protein signaling,

arrestin-mediated signaling, and receptor internalization. A variety of NOPR

ligands, both putative agonists and antagonists, were characterized by

further examination of these ...


Astrocyte-Specific Regulation Of Hmecp2 Expression In Drosophila, David Hess-Homeier, Chia-Yu Fan, Tarun Gupta, Ann-Shyn Chiang, Sarah J. Certel 2014 University of Montana - Missoula

Astrocyte-Specific Regulation Of Hmecp2 Expression In Drosophila, David Hess-Homeier, Chia-Yu Fan, Tarun Gupta, Ann-Shyn Chiang, Sarah J. Certel

Biological Sciences Faculty Publications

Alterations in the expression of Methyl-CpG-binding protein 2 (MeCP2) either by mutations or gene duplication leads to a wide spectrum of neurodevelopmental disorders including Rett Syndrome and MeCP2 duplication disorder. Common features of Rett Syndrome (RTT), MeCP2 duplication disorder, and neuropsychiatric disorders indicate that even moderate changes in MeCP2 protein levels result in functional and structural cell abnormalities. In this study, we investigated two areas of MeCP2 pathophysiology using Drosophila as a model system: the effects of MeCP2 glial gain-of-function activity on circuits controlling sleep behavior, and the cell-type specific regulation of MeCP2 expression. In this study, we first examined ...


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