University of New Mexico 16 articles published in JoVE Immunology and Infection A Flow Cytometry-Based High-Throughput Technique for Screening Integrin-Inhibitory Drugs Ziming Cao1, Matthew J. Garcia2, Larry A. Sklar2,3,4,5, Angela Wandinger-Ness3,4, Zhichao Fan1 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. Bioengineering High Throughput Screening Assessment of Reactive Oxygen Species (ROS) Generation using Dihydroethidium (DHE) Fluorescence Dye Rahul Kumar1,2, Rama R. Gullapalli1,2 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. Medicine Randomized Controlled Trial to Study the Acute Effects of Strength Exercise on Insulin Sensitivity in Obese Adults Luis Filipe Rocha Silva1, Bruna Caroline Chaves Garcia2, Zach A. Mang3, Fabiano Trigueiro Amorim4, Marco Fabrício Dias-Peixoto1, Fernando Gripp1, Valmor Tricoli5, Flávio de Castro Magalhães1,4 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. Environment Visualization of Productivity Zones Based on Nitrogen Mass Balance Model in Narragansett Bay, Rhode Island Jongsun Kim1, Myung Hwangbo1, Patricia S. Thibodeau2, Georgia Rhodes3, Emma Hogarth4, Stewart Copeland3,4 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. Biochemistry An Optimized Single-Molecule Pull-Down Assay for Quantification of Protein Phosphorylation Elizabeth M. Bailey1, Emanuel Salazar-Cavazos1, Rachel M. Grattan1, Michael J. Wester2, David J. Schodt2, Julian A. Rojo1, Keith A. Lidke2,3, Diane S. Lidke1,3 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. Medicine A Real-World High-Intensity Interval Training Protocol for Cardiorespiratory Fitness Improvement Fernando Gripp1,2, Gilton de Jesus Gomes1,2, Ricardo Augusto Leoni De Sousa1,2, Júllia Alves de Andrade3, Ilkilene Pinheiro Queiroz3, Caíque Olegário Diniz Magalhães2, Ricardo Cardoso Cassilhas1,2,3, Flávio de Castro Magalhães1,2,3, Fabiano Trigueiro Amorim4, Marco Fabrício Dias-Peixoto1,2,3 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. Biochemistry Detection of Total Reactive Oxygen Species in Adherent Cells by 2',7'-Dichlorodihydrofluorescein Diacetate Staining Hyeoncheol Kim1, Xiang Xue1 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. Chemistry An Externally-Heated Diamond Anvil Cell for Synthesis and Single-Crystal Elasticity Determination of Ice-VII at High Pressure-Temperature Conditions Xiaojing Lai1,2, Feng Zhu2, Jin S. Zhang3, Dongzhou Zhang2,4, Sergey Tkachev4, Vitali B. Prakapenka4, Bin Chen2 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. Genetics Use of Drosophila S2 Cells for Live Imaging of Cell Division Evan B. Dewey1, Amalia S. Parra1, Christopher A. Johnston1 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. Developmental Biology Isolation of CD 90+ Fibroblast/Myofibroblasts from Human Frozen Gastrointestinal Specimens Paul Johnson1, Ellen J. Beswick3, Celia Chao1, Don W. Powell2, Mark R. Hellmich1, Iryna V. Pinchuk2 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. Medicine Modeling Encephalopathy of Prematurity Using Prenatal Hypoxia-ischemia with Intra-amniotic Lipopolysaccharide in Rats Lauren L. Jantzie1,2, Jesse L. Winer3, Jessie R. Maxwell1, Lindsay A.S. Chan3, Shenandoah Robinson3,4 1Department of Pediatrics, University of New Mexico, 2Department of Neurosciences, University of New Mexico, 3 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. Bioengineering Epithelial Cell Repopulation and Preparation of Rodent Extracellular Matrix Scaffolds for Renal Tissue Development Joseph S. Uzarski1,2, Jimmy Su1,2,3,4, Yan Xie1,2, Zheng J. Zhang1,2, Heather H. Ward5, Angela Wandinger-Ness6, William M. Miller7,8, Jason A. Wertheim1,2,3,4,8,9 1Comprehensive Transplant Center, Feinberg School of Medicine, Northwestern University, 2Department of Surgery, Feinberg School of Medicine, Northwestern University, 3Department of Biomedical Engineering, Northwestern University, 4Simpson Querrey Institute for BioNanotechnology in Medicine, Northwestern University, 5Department of Internal Medicine, University of New Mexico HSC, 6Department of Pathology, University of New Mexico HSC, 7Department of Chemical and Biological Engineering, Northwestern University, 8Chemistry of Life Processes Institute, Northwestern University, 9Department of Surgery, Jesse Brown VA Medical Center This protocol describes decellularization of Sprague Dawley rat kidneys by antegrade perfusion of detergents through the vasculature, producing acellular renal extracellular matrices that serve as templates for repopulation with human renal epithelial cells. Recellularization and use of the resazurin perfusion assay to monitor growth is performed within specially-designed perfusion bioreactors. Behavior A Cognitive Paradigm to Investigate Interference in Working Memory by Distractions and Interruptions Jacki Janowich1, Jyoti Mishra2, Adam Gazzaley2,3,4 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. Environment Removal of Trace Elements by Cupric Oxide Nanoparticles from Uranium In Situ Recovery Bleed Water and Its Effect on Cell Viability Jodi R. Schilz1, K. J. Reddy2, Sreejayan Nair3, Thomas E. Johnson4, Ronald B. Tjalkens5, Kem P. Krueger3, Suzanne Clark6 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. Behavior Moderate Prenatal Alcohol Exposure and Quantification of Social Behavior in Adult Rats Derek A. Hamilton1,2, Christy M. Magcalas1, Daniel Barto1, Clark W. Bird1, Carlos I. Rodriguez1, Brandi C. Fink3, Sergio M. Pellis4, Suzy Davies2, Daniel D. Savage1,2 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. Medicine Construction of Vapor Chambers Used to Expose Mice to Alcohol During the Equivalent of all Three Trimesters of Human Development Russell A. Morton1, Marvin R. Diaz1, Lauren A. Topper1, C. Fernando Valenzuela1 1Department of Neurosciences, School of Medicine, University of New Mexico Health Sciences Center We demonstrate the construction of alcohol vapor chambers using readily available materials that simultaneously house 6 mouse cages. We further describe their use in a mouse model of fetal alcohol exposure equivalent to all 3 trimesters of human pregnancy. This paradigm exposes animals during gestation and postnatal days 1-12.
