University of New Mexico

12 articles published in JoVE

• 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. Lidke^2,3, Diane S. Lidke^{1,3 1}Department 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 Gripp^1,2, Gilton de Jesus Gomes^1,2, Ricardo Augusto Leoni De Sousa^1,2, Júllia Alves de Andrade³, Ilkilene Pinheiro Queiroz³, Caíque Olegário Diniz Magalhães², Ricardo Cardoso Cassilhas^1,2,3, Flávio de Castro Magalhães^1,2,3, Fabiano Trigueiro Amorim⁴, Marco Fabrício Dias-Peixoto^{1,2,3 1}Department 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 Xue^{1 1}Department 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 Lai^1,2, Feng Zhu², Jin S. Zhang³, Dongzhou Zhang^2,4, Sergey Tkachev⁴, Vitali B. Prakapenka⁴, Bin Chen^{2 1}Gemmological 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. Johnston^{1 1}Department 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. Pinchuk^{2 1}Surgery, 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. Jantzie^1,2, Jesse L. Winer³, Jessie R. Maxwell¹, Lindsay A.S. Chan³, Shenandoah Robinson^{3,4 1}Department 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. Uzarski^1,2, Jimmy Su^1,2,3,4, Yan Xie^1,2, Zheng J. Zhang^1,2, Heather H. Ward⁵, Angela Wandinger-Ness⁶, William M. Miller^7,8, Jason A. Wertheim^{1,2,3,4,8,9 1}Comprehensive 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 Gazzaley^{2,3,4 1}Department 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 Clark^{6 1}Division 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. Hamilton^1,2, Christy M. Magcalas¹, Daniel Barto¹, Clark W. Bird¹, Carlos I. Rodriguez¹, Brandi C. Fink³, Sergio M. Pellis⁴, Suzy Davies², Daniel D. Savage^{1,2 1}Department 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 Valenzuela^{1 1}Department 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.