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18 ARTICLES PUBLISHED IN JoVE

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Biology

Imaging In-Stent Restenosis: An Inexpensive, Reliable, and Rapid Preclinical Model
Tobias Deuse 1, Fumiaki Ikeno 2, Robert C. Robbins 1, Sonja Schrepfer 1
1Department of Cardiothoracic Surgery, Stanford University School of Medicine, 2Stanford University School of Medicine

This video demonstrates how to use a preclinical inexpensive and reliable model to study pathobiological and pathophysiological processes of in-stent restenosis development. Longitudinal in vivo monitoring using OCT (Optical Coherence Tomography) and analysis of OCT images are also demonstrated.

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Biology

Heterotopic and Orthotopic Tracheal Transplantation in Mice used as Models to Study the Development of Obliterative Airway Disease
Xiaoqin Hua 1, Tobias Deuse 1,2, Karis R. Tang-Quan 1,2,3, Robert C. Robbins 3, Hermann Reichenspurner 1,2, Sonja Schrepfer 1,2,3
1Transplant and Stem Cell Immunobiology Lab (TSI), University Heart Center Hamburg, 2CVRC, University Hospital Hamburg, 3Department of CT Surgery, Stanford University School of Medicine

This video shows and compares two experimental models to study the development of obliterative airway disease (OAD) in mice, the heterotopic and orthotopic tracheal transplantation model.

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Biology

LAD-Ligation: A Murine Model of Myocardial Infarction
Mandy V.V. Kolk 1,2, Danja Meyberg 1,2, Tobias Deuse 1,2, Karis R. Tang-Quan 1,2,3, Robert C. Robbins 3, Hermann Reichenspurner 1,2, Sonja Schrepfer 1,2,3
1Transplant and Stem Cell Immunobiology Lab (TSI), University Heart Center Hamburg, 2CVRC, University Hospital Hamburg, 3Department of CT Surgery, Stanford University School of Medicine

This video demonstrates how to use a fast and reliable model to study pathobiological and pathophysiological processes of myocardial ischemia.

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Medicine

Orthotopic Aortic Transplantation: A Rat Model to Study the Development of Chronic Vasculopathy
Mandy Stubbendorff 1, Tobias Deuse 1,2, Anna Hammel 1, Robert C. Robbins 2, Hermann Reichenspurner 1, Sonja Schrepfer 1,2
1University Heart Center Hamburg, Transplant and Stem Cell Immunobiology Lab (TSI), University Hospital Hamburg, 2Stanford University School of Medicine

This video demonstrates the orthotopic aortic transplant model as a simple model to study the development of transplant vasculopathy (TVP) in rats.

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Bioengineering

Bioluminescence Imaging for Assessment of Immune Responses Following Implantation of Engineered Heart Tissue (EHT)
Lenard Conradi 1,2, Christiane Pahrmann 1, Stephanie Schmidt 1, Tobias Deuse 1,3, Arne Hansen 2, Alexandra Eder 2, Hermann Reichenspurner 1, Robert C. Robbins 3, Thomas Eschenhagen 2, Sonja Schrepfer 1,3
1Transplant and Stem Cell Immunobiology Lab (TSI) and CVRC, University Hospital Hamburg, University Heart Center Hamburg, 2Department of Experimental and Clinical Pharmacology and Toxicology, University Heart Center Hamburg, 3CT Surgery, Stanford University School of Medicine

This video demonstrates the use of in vivo bioluminescence imaging to study immune responses after implantation of Engineered Heart Tissue (EHT) in rats.

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Medicine

Human Internal Mammary Artery (IMA) Transplantation and Stenting: A Human Model to Study the Development of In-Stent Restenosis
Xiaoqin Hua 1,2, Tobias Deuse 1,2, Evangelos D. Michelakis 3, Alois Haromy 3, Phil S. Tsao 4, Lars Maegdefessel 4, Reinhold G. Erben 5, Claudia Bergow 5, Boris B. Behnisch 6, Hermann Reichenspurner 1,2, Robert C. Robbins 7, Sonja Schrepfer 1,2,7
1University Heart Center Hamburg, TSI-Lab, Germany, 2Cardiovascular Research Center, University of Hamburg, 3Department of Medicine, Cardiology Division, Pulmonary Hypertension Program, University of Alberta, 4Department of Medicine, Stanford University School of Medicine , 5Department of Biomedical Sciences, Institute of Physiology, Pathophysiology, and Biophysics, University of Veterinary Medicine, Vienna, 6Translumina GmbH, Hechingen, 7Department of Cardiothoracic Surgery, Stanford University School of Medicine

This video shows a model to study the development of intimal hyperplasia after stent deployment using a human vessel (IMA) in an immunodeficient rat model.

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Education

Spatial Separation of Molecular Conformers and Clusters
Daniel Horke 1, Sebastian Trippel 1, Yuan-Pin Chang 1, Stephan Stern 1,2, Terry Mullins 1, Thomas Kierspel 1,3, Jochen Küpper 1,2,3
1Center for Free-Electron Laser Science, CFEL, DESY, 2Department of Physics, University of Hamburg, 3The Hamburg Center for Ultrafast Imaging, University of Hamburg

We present a technique that allows the spatial separation of different conformers or clusters present in a molecular beam. An electrostatic deflector is used to separate species by their mass-to-dipole moment ratio, leading to the production of gas-phase ensembles of a single conformer or cluster stoichiometry.

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Medicine

Inducing Myointimal Hyperplasia Versus Atherosclerosis in Mice: An Introduction of Two Valid Models
Mandy Stubbendorff *1,2, Xiaoqin Hua *1,2, Tobias Deuse 1,2,3, Ziad Ali 4,5, Hermann Reichenspurner 2,3, Lars Maegdefessel 6, Robert C. Robbins 7, Sonja Schrepfer 1,2,3,4
1Transplant and Stem Cell Immunobiology Lab, Cardiovascular Research Center, University Hospital Hamburg, 2Cardiovascular Research Center (CVRC) and DZHK University Hamburg, 3Department of Cardiovascular Surgery, University Heart Center Hamburg, 4Center for Interventional Vascular Therapy, Division of Cardiology, Columbia University, 5Cardiovascular Research Foundation, New York, 6Karolinska Institute, Stockholm, 7Department of Cardiothoracic Surgery, Stanford University School of Medicine, Falk Cardiovascular Research Center

This video shows two models of intimal plaque development in murine arteries and emphasizes the differences in myointimal hyperplasia and atherosclerosis.

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JoVE Core

Synthesis of Indoxyl-glycosides for Detection of Glycosidase Activities
Stephan Böttcher 1, Joachim Thiem 1
1Department of Chemistry, Faculty of Sciences, University of Hamburg

Indoxyl glycosides are well-established and widely used tools for enzyme screening and enzyme activity monitoring. Especially for glucose type structures previous syntheses proved to be challenging and low yielding. Our novel approach employs indoxylic acid esters as precious intermediates to yield a considerable number of indoxyl glycosides in good yields.

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Medicine

Vein Interposition Model: A Suitable Model to Study Bypass Graft Patency
Dong Wang 1,2,3,4, Grigol Tediashvili 1,2,3, Simon Pecha 4, Hermann Reichenspurner 4, Tobias Deuse 1,2,3,4, Sonja Schrepfer 1,2,3,4
1Transplant and Stem Cell Immunobiology Lab, University Heart Center Hamburg, 2Department of Surgery, Transplant and Stem Cell Immunobiology Lab, University of California San Francisco (UCSF), 3Cardiovascular Research Center (CVRC) and DZHK German Center for Cardiovascular Research), partner site Hamburg/Kiel/Luebeck, 4Cardiovascular Surgery, University Heart Center Hamburg

This video demonstrates a model to study the development of myointimal hyperplasia after venous interposition surgery in rats.

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Medicine

Balloon-based Injury to Induce Myointimal Hyperplasia in the Mouse Abdominal Aorta
Grigol Tediashvili 1,2,3, Dong Wang 1,2,3,4, Hermann Reichenspurner 4, Tobias Deuse 1,2,3,4, Sonja Schrepfer 1,2,3,4
1Transplant and Stem Cell Immunobiology Lab, University Heart Center, 2Department of Surgery, Transplant and Stem Cell Immunobiology Lab, University of California San Francisco (UCSF), 3Cardiovascular Research Center (CVRC) and DZHK German Center for Cardiovascular Research, 4Cardiovascular Surgery, University Heart Center

This article demonstrates a murine model to study the development of myointimal hyperplasia (MH) after aortic balloon injury.

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Biochemistry

Growing Protein Crystals with Distinct Dimensions Using Automated Crystallization Coupled with In Situ Dynamic Light Scattering
Daniela Baitan 1,2, Robin Schubert 2,3, Arne Meyer 1, Karsten Dierks 1, Markus Perbandt 2,3, Christian Betzel 2,3
1Xtal Concepts GmbH, 2Institute for Biochemistry and Molecular Biology, Laboratory for Structural Biology of Infection and Inflammation c/o DESY, University of Hamburg, 3The Hamburg Center for Ultrafast Imaging, University of Hamburg

Here we present a protocol for controlled production of protein microcrystals. The process uses an automated device allowing controlled manipulation of several crystallization parameters. The protein crystallization is carried-out by controlled and automated addition of crystallization solutions while monitoring and investigating the radius distribution of particles in the crystallization droplet.

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Chemistry

Microfluidic Chips for In Situ Crystal X-ray Diffraction and In Situ Dynamic Light Scattering for Serial Crystallography
Yannig Gicquel *1,2, Robin Schubert *3,4,5, Svetlana Kapis 3, Gleb Bourenkov 6, Thomas Schneider 6, Markus Perbandt 3,4, Christian Betzel 3,4,5, Henry N. Chapman 1,2,4, Michael Heymann 1,7
1Center for Free Electron Laser Science, DESY, 2Department of Physics, University of Hamburg, 3Institute for Biochemistry and Molecular Biology, Laboratory for Structural Biology of Infection and Inflammation, University of Hamburg, 4The Hamburg Center for Ultrafast Imaging, University of Hamburg, 5Integrated Biology Infrastructure Life-Science Facility at the European XFEL (XBI), 6European Molecular Biology Laboratory, EMBL c/o DESY, 7Department of Cellular and Molecular Biophysics, Max Planck Institute of Biochemistry

This protocol describes in detail how to fabricate and operate microfluidic devices for X-ray diffraction data collection at room temperature. Additionally, it describes how to monitor protein crystallization by dynamic light scattering and how to process and analyze obtained diffraction data.

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Medicine

Implantation of hiPSC-derived Cardiac-muscle Patches after Myocardial Injury in a Guinea Pig Model
Liesa Castro 1,2, Birgit Geertz 3, Marina Reinsch 2,3, Bülent Aksehirlioglu 3, Arne Hansen 2,3, Thomas Eschenhagen 2,3, Hermann Reichenspurner 1,2, Florian Weinberger *2,3, Simon Pecha *1,2
1Department of Cardiovascular Surgery, University Heart Center Hamburg, 2partner site Hamburg/Kiel/Lübeck, German Centre for Cardiovascular Research (DZHK), 3Department of Experimental Pharmacology and Toxicology, Cardiovascular ResearchCenter, University Medical Center Hamburg-Eppendorf

Here we present a protocol for the induction of left ventricular cryoinjury followed by the implantation of a cardiac muscle patch, derived from human iPS-cell cardiomyocytes in a guinea pig model.

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Bioengineering

CRISPR/Cas12a Multiplex Genome Editing of Saccharomyces cerevisiae and the Creation of Yeast Pixel Art
Klaudia Ciurkot 1,2, Brenda Vonk 1, Thomas E. Gorochowski 3,4, Johannes A. Roubos 1, René Verwaal 1
1DSM Biotechnology Center, 2Biochemistry and Molecular Biology, Department of Chemistry, University of Hamburg, 3BrisSynBio, University of Bristol, Life Sciences Building, 4School of Biological Sciences, University of Bristol, Life Sciences Building

The CRISPR/Cas12a system in combination with a single crRNA array enables efficient multiplex editing of the S. cerevisiae genome at multiple loci simultaneously. This is demonstrated by constructing carotenoid producing yeast strains which are subsequently used to create yeast pixel art.

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Medicine

A Cryoinjury Model to Study Myocardial Infarction in the Mouse
Dong Wang *1,2, Grigol Tediashvili *1,2, Xiaomeng Hu 1,2, Alessia Gravina 2, Sivan G. Marcus 1,2, Hao Zhang 4, Jeffrey E Olgin 4, Tobias Deuse 1,2,5, Sonja Schrepfer 1,2,3,5
1Transplant and Stem Cell Immunobiology Lab, University Heart Center, 2Department of Surgery, Transplant and Stem Cell Immunobiology Lab, University of California San Francisco, 3Cardiovascular Research Center (CVRC) and DZHK German Center for Cardiovascular Research, 4Division of Cardiology, Cardiovascular Research Institute, University of California San Francisco, 5Cardiovascular Surgery, University Heart Center

This article demonstrates a model to study cardiac remodeling after myocardial cryoinjury in mice.

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Immunology and Infection

Immunofluorescence Imaging of Neutrophil Extracellular Traps in Human and Mouse Tissues
Lavinia Schoenfeld 1, Birgit Appl 1, Laia Pagerols-Raluy 1, Annika Heuer 3, Konrad Reinshagen 1, Michael Boettcher 1,2
1Department of Pediatric Surgery, University Medical Center Hamburg-Eppendorf, University of Hamburg, 2Department of Pediatric Surgery, University Medical Center Mannheim, University of Heidelberg, 3Division of Spine Surgery, Department of Trauma and Orthopedic Surgery, University Medical Center Hamburg-Eppendorf (UKE)

Neutrophil extracellular traps (NETs) are associated with various diseases, and immunofluorescence is often used for their visualization. However, there are various staining protocols, and, in many cases, only one type of tissue is examined. Here, we establish a generally applicable protocol for staining NETs in mouse and human tissue.

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Biochemistry

Design and Construction of an Experimental Setup to Enhance Mineral Weathering through the Activity of Soil Organisms
Tullia Calogiuri 1,2, Mathilde Hagens 2, Jan Willem Van Groenigen 1, Thomas Corbett 3, Jens Hartmann 4, Rick Hendriksen 5, Iris Janssens 6, Ivan A. Janssens 7, Guillermo Ledesma Dominguez 7, Grant Loescher 4, Steven Mortier 6, Anna Neubeck 3, Harun Niron 7, Reinaldy P. Poetra 4, Lukas Rieder 4, Eric Struyf 7, Michiel Van Tendeloo 8, Tom De Schepper 6, Tim Verdonck 9, Siegfried E. Vlaeminck 8, Sara Vicca 7, Alix Vidal 1
1Soil Biology Group, Wageningen University & Research, 2Soil Chemistry and Chemical Soil Quality, Wageningen University & Research, 3Department of Earth Sciences, Uppsala University, 4Institute for Geology, Center for Earth System Research and Sustainability, University of Hamburg, 5Tupola, Wageningen University & Research, 6IDLab - Department of Computer Science, University of Antwerp - imec, 7Plants and Ecosystems (PLECO), Biology Department, University of Antwerp, 8Research Group of Sustainable Energy, Air and Water Technology, University of Antwerp, 9Department of Mathematics, University of Antwerp - imec

Here we present the construction and operation of an experimental setup to enhance mineral weathering through the activity of soil organisms while concurrently manipulating abiotic variables known to stimulate weathering. Representative results from the functioning of the setup and sample analyses are discussed together with points for improvement.

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