University of Colorado, Denver

8 ARTICLES PUBLISHED IN JoVE

Medicine

Pressure Controlled Ventilation to Induce Acute Lung Injury in Mice

Michael Koeppen¹, Tobias Eckle¹, Holger K. Eltzschig¹

¹Department of Anesthesiology, University of Colorado

A murine model for ventilator induced lung injury is an important tool to study an acute lung injury in vivo. Here, we report an easy applicable in situ model for acute lung injury using high-pressure mechanical ventilation to induce acute failure of the lung.

Medicine

Use of a Hanging-weight System for Isolated Renal Artery Occlusion

Almut Grenz¹, Julee H. Hong¹, Alexander Badulak¹, Douglas Ridyard¹, Timothy Luebbert², Jae-Hwan Kim³, Holger K. Eltzschig¹

¹Department of Anesthesiology, University of Colorado, ²School of Medicine, University of Colorado, ³Department of Anesthesiology, Korea University College of Medicine

A precise murine model for acute kidney injury (AKI) due to ischemia is an important tool to investigate acute kidney injury and possibly find therapeutic tools to treat renal injury. The hanging weight system offers a tool for immediate and reliable renal artery occlusion and reperfusion without causing renal congestion.

Medicine

Use of a Hanging-weight System for Liver Ischemia in Mice

Michael Zimmerman¹, Eunyoung Tak², Maria Kaplan¹, Mercedes Susan Mandell², Holger K. Eltzschig², Almut Grenz²

¹UCH Transplant Center, University of Colorado, Denver, ²Department of Anesthesiology, University of Colorado, Denver

We established a novel murine model of a hanging weight system for portal triad occlusion. This technique may be useful for future investigations of ischemia in murine hepatic models.

Medicine

Assessment of Vascular Function in Patients With Chronic Kidney Disease

Kristen L. Jablonski¹, Emily Decker¹, Loni Perrenoud¹, Jessica Kendrick¹, Michel Chonchol¹, Douglas R. Seals², Diana Jalal¹

¹Division of Renal Diseases and Hypertension, University of Colorado, Denver, ²Department of Integrative Physiology, University of Colorado, Boulder

The degree of vascular dysfunction and contributing physiological mechanisms can be assessed in patients with chronic kidney disease by measuring brachial artery flow-mediated dilation, aortic pulse-wave velocity, and vascular endothelial cell protein expression.

Biology

Highly Efficient Ligation of Small RNA Molecules for MicroRNA Quantitation by High-Throughput Sequencing

Jerome E. Lee¹, Rui Yi^1,2

¹Molecular, Cellular, and Developmental Biology, University of Colorado, Boulder, ²Linda Crnic Institute for Down Syndrome, University of Colorado, Denver

MicroRNAs (miRNAs) are a widely conserved class of regulatory molecules. Here we describe a miRNA cloning method that relies upon two potent ligation steps followed by high-throughput sequencing. Our method permits accurate genome-wide quantitation of miRNAs.

Medicine

Murine Kidney Transplant Technique

Robert Plenter^1,2, Swati Jain³, Chelsea M. Ruller⁴, Trevor L. Nydam⁴, Alkesh H. Jani^3,5

¹Colorado Center for Transplantation Care, Research and Education, University of Colorado, Denver, ²Division of Pulmonary Sciences and Critical Care Medicine, University of Colorado, Denver, ³Department of Medicine, Division of Renal Diseases and Hypertension, Medical Center and University of Colorado, Denver, ⁴Department of Surgery, Division of Transplant Surgery, University of Colorado School of Medicine, University of Colorado-Denver, ⁵Renal Section, Denver Veterans Affairs Medical Center

The goal of this manuscript is to describe the steps required to perform a kidney transplant in a mouse, paying particular attention to the details of the arterial anastomosis.

Chemistry

Protocol for the Solid-phase Synthesis of Oligomers of RNA Containing a 2'-O-thiophenylmethyl Modification and Characterization via Circular Dichroism

Andrew J. Francis¹, Marino J. E. Resendiz¹

¹Department of Chemistry, University of Colorado Denver

This article provides a detailed procedure on the solid-phase synthesis, purification, and characterization of dodecamers of RNA modified at the C2'-O-position. UV-vis and circular dichroism photometric analyses are used to quantify and characterize structural aspects, i.e., single-strands or double-strands.

JoVE Journal

Identification of RNA Fragments Resulting from Enzymatic Degradation using MALDI-TOF Mass Spectrometry

Shawn W. Schowe¹, Conner J. Langeberg², Erich G. Chapman², Kitty Brown³, Marino J. E. Resendiz¹

¹Department of Chemistry, University of Colorado, Denver, ²Department of Chemistry & Biochemistry, University of Denver, ³Analytical Resources Core Bioanalysis & Omics, Colorado State University

MALDI-TOF was used to characterize fragments obtained from the reactivity between oxidized RNA and the exoribonuclease Xrn-1. The present protocol describes a methodology that can be applied to other processes involving RNA and/or DNA.