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University of New Mexico

17 ARTICLES PUBLISHED IN JoVE

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Medicine

In vitro Organoid Culture of Primary Mouse Colon Tumors
Xiang Xue 1, Yatrik M. Shah 1,2
1Department of Molecular & Integrative Physiology, University of Michigan , 2Department of Internal Medicine, Division of Gastroenterology, University of Michigan

A simple method to establish primary murine colon tumor organoid is described. This method utilizes the feature that colon tumor cells survive and grow into organoids in media containing limited growth factors, whereas normal colon epithelial do not.

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Behavior

A Cognitive Paradigm to Investigate Interference in Working Memory by Distractions and Interruptions
Jacki Janowich 1, Jyoti Mishra 2, Adam Gazzaley 2,3,4
1Department of Psychology, University of New Mexico, 2Department of Neurology, University of California, San Francisco, 3Department of Physiology, Center for Integrative Neuroscience, University of California, San Francisco, 4Department of Psychiatry, University of California, San Francisco

A novel cognitive paradigm is developed to elucidate behavioral and neural correlates of interference by to-be-ignored distractors versus interference by to-be-attended interruptors during a working memory task. In this manuscript, several variants of this paradigm are detailed, and data obtained with this paradigm in younger/older adult participants is reviewed.

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Behavior

Moderate Prenatal Alcohol Exposure and Quantification of Social Behavior in Adult Rats
Derek A. Hamilton 1,2, Christy M. Magcalas 1, Daniel Barto 1, Clark W. Bird 1, Carlos I. Rodriguez 1, Brandi C. Fink 3, Sergio M. Pellis 4, Suzy Davies 2, Daniel D. Savage 1,2
1Department of Psychology, University of New Mexico, 2Department of Neurosciences, University of New Mexico, 3Department of Psychiatry and Behavioral Sciences, University of New Mexico, 4Canadian Centre for Behavioural Neuroscience, University of Lethbridge

The goal of the protocol presented here is to describe procedures to expose rats to moderate levels of alcohol during prenatal brain development and to quantify resulting alterations in social behavior during adulthood.

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Environment

Removal of Trace Elements by Cupric Oxide Nanoparticles from Uranium In Situ Recovery Bleed Water and Its Effect on Cell Viability
Jodi R. Schilz 1, K. J. Reddy 2, Sreejayan Nair 3, Thomas E. Johnson 4, Ronald B. Tjalkens 5, Kem P. Krueger 3, Suzanne Clark 6
1Division of Physical Therapy, Department of Orthopedics & Rehabilitation, University of New Mexico, 2Department of Ecosystem Science and Management, University of Wyoming, 3School of Pharmacy, University of Wyoming, 4Department of Environmental and Radiological Health Sciences, Colorado State University, 5Center for Environmental Medicine, Colorado State University, 6College of Pharmacy, California Northstate University

Production bleed water (PBW) was treated with cupric oxide nanoparticles (CuO-NPs) and cellular toxicity was assessed in cultured human cells. The goal of this protocol was to integrate the native environmental sample into a cell culture format assessing the changes in toxicity due to CuO-NP treatment.

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Medicine

Modeling Encephalopathy of Prematurity Using Prenatal Hypoxia-ischemia with Intra-amniotic Lipopolysaccharide in Rats
Lauren L. Jantzie 1,2, Jesse L. Winer 3, Jessie R. Maxwell 1, Lindsay A.S. Chan 3, Shenandoah Robinson 3,4
1Department of Pediatrics, University of New Mexico, 2Department of Neurosciences, University of New Mexico, 3Department of Neurosurgery, Boston Children's Hospital, 4Department of Neurology, Harvard Medical School

Encephalopathy of prematurity encompasses the central nervous system abnormalities associated with injury from preterm birth. This report describes a clinically relevant rat model of in utero transient systemic hypoxia-ischemia and intra-amniotic lipopolysaccharide administration (LPS) that mimics chorioamnionitis, and the related impact of infectious stimuli and placental underperfusion on CNS development.

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Developmental Biology

Isolation of CD 90+ Fibroblast/Myofibroblasts from Human Frozen Gastrointestinal Specimens
Paul Johnson 1, Ellen J. Beswick 3, Celia Chao 1, Don W. Powell 2, Mark R. Hellmich 1, Iryna V. Pinchuk 2
1Surgery, University of Texas Medical Branch, 2Internal Medicine, University of Texas Medical Branch, 3Molecular Genetics and Microbiology, University of New Mexico

Here, a protocol to isolate and establish primary fibroblast/myofibroblast (MF) cultures from frozen gastric, small intestinal, and colonic tissue-yielding cells with a MF phenotype-is presented. These cells express CD90, α-SMA and vimentin. MFs can be used for a variety of functional assays including enzymatic activity and cytokine production.

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Chemistry

High Pressure Single Crystal Diffraction at PX^2
Dongzhou Zhang 1, Przemyslaw K. Dera 1, Peter J. Eng 2, Joanne E. Stubbs 2, Jin S. Zhang 1, Vitali B. Prakapenka 2, Mark L. Rivers 2
1Hawai’i Institute of Geophysics and Planetology, University of Hawai’i at Manoa, 2Center for Advanced Radiation Sources, University of Chicago

In this report, we describe detailed procedures for carrying out single crystal X-ray diffraction experiments with a diamond anvil cell at the GSECARS 13-BM-C beamline at the Advanced Photon Source. ATREX and RSV programs are used to analyze the data.

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JoVE Core

Use of Drosophila S2 Cells for Live Imaging of Cell Division
Evan B. Dewey 1, Amalia S. Parra 1, Christopher A. Johnston 1
1Department of Biology, University of New Mexico

Cell divisions can be visualized in real time using fluorescently tagged proteins and time-lapse microscopy. Using the protocol presented here, users can analyze cell division timing dynamics, mitotic spindle assembly, and chromosome congression and segregation. Defects in these events following RNA interference (RNAi)-mediated gene knockdown can be assessed and quantified.

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Biochemistry

Detection of Total Reactive Oxygen Species in Adherent Cells by 2',7'-Dichlorodihydrofluorescein Diacetate Staining
Hyeoncheol Kim 1, Xiang Xue 1
1Department of Biochemistry and Molecular Biology, University of New Mexico

Here, we present a protocol to detect total cellular reactive oxygen species (ROS) using 2',7'-dichlorodihydrofluorescein diacetate (DCFH-DA). This method can visualize cellular ROS localization in adherent cells with a fluorescence microscope and quantify ROS intensity with a fluorescence plate reader. This protocol is simple, efficient and cost-effective.

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Chemistry

An Externally-Heated Diamond Anvil Cell for Synthesis and Single-Crystal Elasticity Determination of Ice-VII at High Pressure-Temperature Conditions
Xiaojing Lai 1,2, Feng Zhu 2, Jin S. Zhang 3, Dongzhou Zhang 2,4, Sergey Tkachev 4, Vitali B. Prakapenka 4, Bin Chen 2
1Gemmological Institute, China University of Geosciences (Wuhan), 2Hawai'i Institute of Geophysics and Planetology, University of Hawai'i at Mānoa, 3Department of Earth and Planetary Sciences, University of New Mexico, 4Center for Advanced Radiation Sources, University of Chicago

This work focuses on the standard protocol for preparing the externally-heated diamond anvil cell (EHDAC) for generating high-pressure and high-temperature (HPHT) conditions. The EHDAC is employed to investigate materials in Earth and planetary interiors under extreme conditions, which can be also used in solid state physics and chemistry studies.

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Biology

Non-thermal Infrared Light Treatment of Ischemia/Reperfusion Injury and Subsequent Analysis of Macrophage Differentiation
Tyler Compton 1, Nicholas Poellinger 2, Janine Struve 3, John G. Krolikowski 1, James T. Ninomiya 4, Dorothee Weihrauch 1
1Department of Anesthesiology, Medical College of Wisconsin, 2Department of Internal Medicine, Loyola University Medical Center, 3Department of Orthopedic Surgery, Medical College of Wisconsin, 4Department of Orthopedic Surgery, University of New Mexico

We describe the reduction of reperfusion injury by 670 nm irradiation in a mouse model of ischemia and reperfusion by tourniquet placement. This 670 nm irradiation reduced the inflammatory response, decreased the number of proinflammatory macrophages, and increased the protective macrophages.

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Biochemistry

An Optimized Single-Molecule Pull-Down Assay for Quantification of Protein Phosphorylation
Elizabeth M. Bailey 1, Emanuel Salazar-Cavazos 1, Rachel M. Grattan 1, Michael J. Wester 2, David J. Schodt 2, Julian A. Rojo 1, Keith A. Lidke 2,3, Diane S. Lidke 1,3
1Department of Pathology, University of New Mexico School of Medicine, 2Department of Physics and Astronomy, University of New Mexico, 3Comprehensive Cancer Center, University of New Mexico Health Sciences Center

The present protocol describes sample preparation and data analysis to quantify protein phosphorylation using an improved single-molecule pull-down (SiMPull) assay.

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Medicine

A Real-World High-Intensity Interval Training Protocol for Cardiorespiratory Fitness Improvement
Fernando Gripp 1,2, Gilton de Jesus Gomes 1,2, Ricardo Augusto Leoni De Sousa 1,2, Júllia Alves de Andrade 3, Ilkilene Pinheiro Queiroz 3, Caíque Olegário Diniz Magalhães 2, Ricardo Cardoso Cassilhas 1,2,3, Flávio de Castro Magalhães 1,2,3, Fabiano Trigueiro Amorim 4, Marco Fabrício Dias-Peixoto 1,2,3
1Department of Physical Education, Federal University of the Jequitinhonha and Mucuri Valleys, 2Multicenter Graduate Program in Physiological Sciences, Federal University of the Jequitinhonha and Mucuri Valleys, 3Graduate Program in Health Sciences, Federal University of the Jequitinhonha and Mucuri Valleys, 44Department of Health, Exercise, and Sports Sciences, University of New Mexico

This study presents a low-cost and easy-to-implement "real world" high-intensity interval training (HIIT) protocol for scientific research and discusses its efficiency for cardiorespiratory fitness.

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Immunology and Infection

A Flow Cytometry-Based High-Throughput Technique for Screening Integrin-Inhibitory Drugs
Ziming Cao 1, Matthew J. Garcia 2, Larry A. Sklar 2,3,4,5, Angela Wandinger-Ness 3,4, Zhichao Fan 1
1Department of Immunology, School of Medicine, UConn Health, 2Center for Molecular Discovery, University of New Mexico Health Sciences Center, 3Comprehensive Cancer Center, University of New Mexico Health Sciences Center, 4Department of Pathology, University of New Mexico Health Sciences Center, 5Autophagy, Inflammation, & Metabolism (AIM) Center, University of New Mexico

This protocol describes a flow cytometry-based, high-throughput screening method to identify small-molecule drugs that inhibit β2 integrin activation on human neutrophils.

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Medicine

Randomized Controlled Trial to Study the Acute Effects of Strength Exercise on Insulin Sensitivity in Obese Adults
Luis Filipe Rocha Silva 1, Bruna Caroline Chaves Garcia 2, Zach A. Mang 3, Fabiano Trigueiro Amorim 4, Marco Fabrício Dias-Peixoto 1, Fernando Gripp 1, Valmor Tricoli 5, Flávio de Castro Magalhães 1,4
1Department of Physical Education, Federal University of the Jequitinhonha and Mucuri Valleys, 2Laboratory of Exercise Biology and Immunometabolism, Centro Integrado de Pós-Graduação e Pesquisa em Saúde, Programa Multicêntrico de Pós-Graduação em Ciências Fisiológicas, Federal University of the Jequitinhonha and Mucuri Valleys, 3Occupational Safety & Health, Los Alamos National Laboratory, 4Department of Health, Exercise, and Sports Sciences, University of New Mexico, 5School of Physical Education and Sport, University of Sao Paulo

This study describes a randomized controlled trial protocol aiming at assessing the acute effects of strength exercise volume on insulin sensitivity in obese individuals.

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Environment

Visualization of Productivity Zones Based on Nitrogen Mass Balance Model in Narragansett Bay, Rhode Island
Jongsun Kim 1, Myung Hwangbo 1, Patricia S. Thibodeau 2, Georgia Rhodes 3, Emma Hogarth 4, Stewart Copeland 3,4
1School of Earth, Environmental and Marine Sciences, University of Texas - Rio Grande Valley, 2Graduate School of Oceanography, University of Rhode Island, 3Rhode Island School of Design, 4Department of Art, University of New Mexico

Here, we aim to visualize the zonation of biological productivity in Narragansett Bay, Rhode Island, based on the nitrogen mass balance model. The results will inform nutrient management in the coastal regions to reduce hypoxia and eutrophication.

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Bioengineering

High Throughput Screening Assessment of Reactive Oxygen Species (ROS) Generation using Dihydroethidium (DHE) Fluorescence Dye
Rahul Kumar 1,2, Rama R. Gullapalli 1,2
1Department of Pathology, University of New Mexico, 2Department of Chemical and Biological Engineering, University of New Mexico

This protocol describes a novel method to quantify intracellular reactive oxygen species (ROS) using dihydroethidium (DHE) as a fluorescence dye probe using a high-throughput screening approach. The protocol describes the methods for quantitative assessment of intracellular reactive oxygen species (ROS) in the three different hepatocellular carcinoma cell lines.

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