University of Southern Denmark

12 articles published in JoVE

• Biochemistry

  
  Quantification of Subcellular Glycogen Distribution in Skeletal Muscle Fibers using Transmission Electron Microscopy Rasmus Jensen¹, Niels Ørtenblad², Cristiano di Benedetto³, Klaus Qvortrup³, Joachim Nielsen^{2 1}Research center for applied health science, University College South Denmark, ²Department of Sports Science and Clinical Biomechanics, University of Southern Denmark, ³Department of Biomedical Sciences, Core Facility for Integrated Microscopy, University of Copenhagen A modified post-fixation procedure increases the contrast of glycogen particles in tissue. This paper provides a step-by-step protocol describing how to handle the tissue, conduct the imaging, and use stereological methods to obtain unbiased and quantitative data on fiber type-specific subcellular glycogen distribution in skeletal muscle.

• Medicine

  
  Isolation of Adipose Derived Regenerative Cells for the Treatment of Erectile Dysfunction Following Radical Prostatectomy Sabrina T. Hansen^1,2, Charlotte H. Jensen^2,3,4, Jens A. Sørensen^2,4,5, Søren P. Sheikh^2,3,4, Lars Lund^{1,2,4 1}Department of Urology, Odense University Hospital, ²Clinical Institute, University of Southern Denmark, ³Laboratory of Molecular and Cellular Cardiology, Department of Clinical Biochemistry and Pharmacology, Odense University Hospital, ⁴Center for Vascular Regeneration, Odense University Hospital, ⁵Research Unit for Plastic Surgery, Odense University Hospital Precise disclosure of methods and protocols is crucial for large scale uptake of stem cell therapies. Here, we present a protocol to isolate adipose-derived regenerative cells, used for a single intracavernous injection as treatment of erectile dysfunction (ED) following radical prostatectomy (RP).

• Medicine

  
  Direct Drug Delivery to Kidney via the Renal Artery Ulrika Dahlqvist¹, Tajana Tesan Tomic², Magnus Söderberg³, Jane Stubbe⁴, Camilla Enggaard⁴, Anette Ericsson⁵, Alex-Xianghua Zhou¹, Anna Björnson Granqvist¹, Lena William-Olsson^{1 1}Bioscience Renal, Research and Early Development, Cardiovascular, Renal and Metabolism, BioPharmaceuticals R&D, ²Cell/mol Pharmacology, Research and Early Development, Cardiovascular, Renal and Metabolism, BioPharmaceuticals R&D, ³CVRM Safety, Safety, Clinical Pharmacology & Safety Sciences, R&D, ⁴Department of Cardiovascular and Renal Research, University of Southern Denmark, ⁵Clinical Renal, Late-stage Development, Cardiovascular, Renal and Metabolism, BioPharmaceuticals R&D This manuscript describes a method for targeted delivery to a single kidney via a catheter placed in the infrarenal abdominal aorta in the mouse.

• Medicine

  
  A Revised Method for Inducing Secondary Lymphedema in the Hindlimb of Mice Alexander Wiinholt^1,2, Mads G. Jørgensen², Amar Bučan^1,2, Farima Dalaei^1,2, Jens A. Sørensen^{2 1}University of Southern Denmark, ²Department of Plastic Surgery, Odense University Hospital This animal model enables researchers to induce statistically significant secondary lymphedema in the hindlimb of mice, lasting at least 8 weeks. The model can be used to study the pathophysiology of lymphedema and to investigate novel treatment options.

• Immunology and Infection

  
  Bacterial Peptide Display for the Selection of Novel Biotinylating Enzymes Jeff Granhøj¹, Henrik Dimke^1,2, Per Svenningsen^{1 1}Department of Molecular Medicine, University of Southern Denmark, ²Department of Nephrology, Odense University Hospital Here we present a method to select for novel variants of the E. coli biotin-protein ligase BirA that biotinylates a specific target peptide. The protocol describes the construction of a plasmid for the bacterial display of the target peptide, generation of a BirA library, selection and characterization of BirA variants.

• Immunology and Infection

  
  Quantification of Atherosclerosis in Mice Monica Centa¹, Daniel F.J. Ketelhuth^1,2, Stephen Malin¹, Anton Gisterå^{1 1}Cardiovascular Medicine Unit, Center for Molecular Medicine, Department of Medicine, Karolinska Institutet, Karolinska University Hospital, ²Department of Cardiovascular and Renal Research, Institute for Molecular Medicine, University of Southern Denmark (SDU) Murine models of atherosclerosis are useful tools to investigate pathogenic pathways on a molecular level, but require standardized quantification of lesion development. This protocol describes an optimized method to determine lesion size in the major arterial vessels including the aortic root, aortic arch, and brachiocephalic artery.

• Bioengineering

  
  Uptake of New Lipid-coated Nanoparticles Containing Falcarindiol by Human Mesenchymal Stem Cells Emma Pipó-Ollé¹, Prasad Walke², Martine K. Notabi¹, Rime B. El-Houri¹, Morten Østergaard Andersen¹, David Needham^2,3,4, Eva C. Arnspang^{1 1}Department of Chemical Engineering, Biotechnology and Environmental Technology, University of Southern Denmark, ²Center for Single Particle Science and Engineering (SPSE), Institute for Molecular Medicine, Health Sciences, University of Southern Denmark, ³Department of Mechanical Engineering and Material Science, Duke University, ⁴School of Pharmacy, University of Nottingham This article describes the encapsulation of falcarindiol in lipid-coated 74 nm nanoparticles. The cellular uptake of the nanoparticles by human stem cells into lipid droplets is monitored by fluorescent and confocal imaging. Nanoparticles are fabricated by the rapid injection method of solvent shifting, and their size is measured with the dynamic light scattering technique.

• Biochemistry

  
  Site-Directed Mutagenesis for In Vitro and In Vivo Experiments Exemplified with RNA Interactions in Escherichia Coli Patrick Rosendahl Andreassen¹, Jens Sivkær Pettersen¹, Mikkel Jørgensen^{1 1}Department of Biochemistry and Molecular Biology, University of Southern Denmark Site-directed mutagenesis is a technique used to introduce specific mutations in deoxyribonucleic acid (DNA). This protocol describes how to do site-directed mutagenesis with a 2-step and 3-step polymerase chain reaction (PCR) based approach, which is applicable to any DNA fragment of interest.

• Immunology and Infection

  
  Rapid, Safe, and Simple Manual Bedside Nucleic Acid Extraction for the Detection of Virus in Whole Blood Samples Maiken W. Rosenstierne¹, Christopher E. Jensen¹, Anders Fomsgaard^{1,2 1}Virus Research & Development Laboratory, Statens Serum Institut, ²Infectious Disease Research Unit, University of Southern Denmark Here, we present a protocol for the rapid virus nucleic acid extraction from the virus-inactivated whole blood. The extraction is performed directly in the blood collection tubes and requires no equipment or electricity. The method is not dependent on laboratory facilities and can be used anywhere (e.g., in field hospitals).

• Medicine

  
  Cavernous Nerve Stimulation and Recording of Intracavernous Pressure in a Rat Morten Hox¹, Travis Mann-Gow², Lars Lund^3,4, Peter Zvara^{1,4 1}Biomedical Laboratory and the Research Unit of Urology, Department of Clinical Research, University of Southern Denmark, ²Department of Surgery, University of Vermont, ³Research Unit of Urology, Department of Clinical Research, University of Southern Denmark, ⁴Department of Urology, Odense University Hospital This study describes a simplified surgical procedure and technique for performing cavernous nerve stimulation with the isolation of the nerve-electrode complex using silicone glue and intracavernous pressure measurement.

• Bioengineering

  
  Assessing Collagen and Elastin Pressure-dependent Microarchitectures in Live, Human Resistance Arteries by Label-free Fluorescence Microscopy Maria Bloksgaard¹, Bjarne Thorsted², Jonathan R. Brewer², Jo G. R. De Mey^{1,3 1}Department of Cardiovascular and Renal Research, Institute of Molecular Medicine, University of Southern Denmark, ²Department of Biochemistry and Molecular Biology, University of Southern Denmark, ³Department of Cardiac, Thoracic and Vascular Surgery, Odense University Hospital We describe simultaneous mechanical testing and 3D-imaging of the arterial wall of isolated, live human resistance arteries, and Fiji and Ilastik image analyses for the quantification of elastin and collagen spatial organization and volume densities. We discuss the use of these data in mathematical models of arterial wall mechanics.

• Medicine

  
  Evaluating the Procedure for Performing Awake Cystometry in a Mouse Model Travis K. Mann-Gow*¹, Troy R. Larson*¹, Chrissie T. Wøien², Thomas M. Andersen², Karl-Erik Andersson^3,4, Peter Zvara^{1,2 1}Department of Surgery, University of Vermont, ²Department of Urology and Biomedical Laboratory, University of Southern Denmark, ³Wake Forest Institute for Regenerative Medicine, Wake Forest University, ⁴Institute for Clinical Medicine, Aarhus University This study describes the surgical procedures and experimental techniques for performing awake cystometry in a freely moving mouse. In addition, it provides experimental evidence to support its optimization and standardization.