Anmelden

University of Miami Miller School of Medicine

18 ARTICLES PUBLISHED IN JoVE

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Biology

Mutagenesis and Analysis of Genetic Mutations in the GC-rich KISS1 Receptor Sequence Identified in Humans with Reproductive Disorders
Luciana Madeira da Silva 1, Lauren Vandepas 1, Suzy D.C. Bianco 1,2
1Division of Endocrinology and Metabolism, Department of Medicine, University of Miami Miller School of Medicine, 2Department of Molecular and Cellular Pharmacology, University of Miami Miller School of Medicine

Mutations in the kisspeptin receptor (KISS1R) are associated with reproductive disorders in patients. Here we describe how to introduce mutations of interest in the GC-rich sequence of KISS1R as well as the use of KISS1R constructs to characterize the degradation pathway of the receptor by immunoprecipitation and western blot.

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Medicine

Transplantation into the Anterior Chamber of the Eye for Longitudinal, Non-invasive In vivo Imaging with Single-cell Resolution in Real-time
Midhat H. Abdulreda 1,2, Alejandro Caicedo 1,3,4, Per-Olof Berggren 1,5
1Diabetes Research Institute, University of Miami Miller School of Medicine, 2Department of Surgery, University of Miami Miller School of Medicine, 3Department of Medicine, University of Miami Miller School of Medicine, 4Department of Physiology & Biophysics, University of Miami Miller School of Medicine, 5The Rolf Luft Research Center for Diabetes and Endocrinology, Karolinska Institutet

A new approach combining intraocular transplantation and confocal microscopy enables longitudinal, non-invasive real-time imaging with single-cell resolution within grafted tissues in vivo. We demonstrate how to transplant pancreatic islets into the anterior chamber of the mouse eye.

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Behavior

Development of a Virtual Reality Assessment of Everyday Living Skills
Stacy A. Ruse 1, Vicki G. Davis 1, Alexandra S. Atkins 1, K. Ranga R. Krishnan 2,3, Kolleen H. Fox 4, Philip D. Harvey 5, Richard S.E. Keefe 1,3
1NeuroCog Trials, Inc., 2Duke-NUS Graduate Medical Center, 3Duke University Medical Center, 4Fox Evaluation and Consulting, PLLC, 5University of Miami Miller School of Medicine

A challenge for proving treatment efficacy for cognitive impairments in schizophrenia is finding the optimizing measurement of skills related to everyday functioning.  The Virtual Reality Functional Capacity Assessment Tool (VRFCAT) is an interactive gaming based computerized measure aimed at skills associated with everyday functioning, including baseline impairments and treatment related changes.

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Medicine

Rat Model of Photochemically-Induced Posterior Ischemic Optic Neuropathy
Yan Wang 1,3,4, Dale P. Brown 1, Brant D. Watson 2, Jeffrey L. Goldberg 1,3
1Bascom Palmer Eye Institute, University of Miami Miller School of Medicine, 2Departments of Neurology and Biomedical Engineering, University of Miami Miller School of Medicine, 3Shiley Eye Center, University of California, 4Department of Ophthalmology and Vision Science, Fudan University

The goal of this protocol is to photochemically induce ischemic injury to the posterior optic nerve in rat. This model is critical to studies of the pathophysiology of posterior ischemic optic neuropathy, and therapeutic approaches for this and other optic neuropathies, as well as of other CNS ischemic diseases.

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Medicine

Transplantation of Schwann Cells Inside PVDF-TrFE Conduits to Bridge Transected Rat Spinal Cord Stumps to Promote Axon Regeneration Across the Gap
Yee-Shuan Lee 1, Siliang Wu 2, Treena Livingston Arinzeh 3, Mary Bartlett Bunge 1,4,5
1The Miami Project to Cure Paralysis, University of Miami Miller School of Medicine, 2Department of Materials Science and Engineering, New Jersey Institute of Technology, 3Department of Biomedical Engineering, New Jersey Institute of Technology, 4Department of Cell Biology, University of Miami Miller School of Medicine, 5Department of Neurological Surgery, University of Miami Miller School of Medicine

This article describes a technique to insert a hollow conduit between the spinal cord stumps after complete transection and fill with Schwann cells (SCs) and injectable basement membrane matrix in order to bridge and promote axon regeneration across the gap.

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Behavior

Utilizing the Modified T-Maze to Assess Functional Memory Outcomes After Cardiac Arrest
Celeste Y. C. Wu 1,2, Francesca M. Lerner 4, Alexandre Couto e Silva 3, Harlee E. Possoit 1,2, Tsung-Han Hsieh 1,2, Jake T. Neumann 5, Alireza Minagar 1, Hung Wen Lin 1,2,3, Reggie H. C. Lee 1,2
1Department of Neurology, Louisiana State University Health Science Center, 2Center for Brain Health, Louisiana State University Health Science Center, 3Department of Cellular Biology and Anatomy, Louisiana State University Health Science Center, 4Department of Neurology, Cerebral Vascular Disease Research Laboratories, University of Miami Miller School of Medicine, 5Department of Biomedical Sciences, West Virginia School of Osteopathic Medicine

This protocol describes the use of a modified T-maze to evaluate functional learning/memory in asphyxia cardiac arrest-induced cerebral ischemia.

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Medicine

Generating a Murine Orthotopic Metastatic Breast Cancer Model and Performing Murine Radical Mastectomy
Eriko Katsuta 1, Masanori Oshi 1, Omar M. Rashid 2,3,4,5, Kazuaki Takabe 1,6,7,8,9,10
1Breast Surgery, Department of Surgical Oncology, Roswell Park Comprehensive Cancer Center, 2Holy Cross Hospital Michael and Dianne Bienes Comprehensive Cancer Center, 3Department of Surgery, Massachusetts General Hospital, 4Department of Surgery, University of Miami Miller School of Medicine, 5Department of Surgery, Nova Southeastern University School of Medicine, 6Department of Surgery, University at Buffalo Jacobs School of Medicine and Biomedical Sciences, 7Department of Breast Surgery and Oncology, Tokyo Medical University, 8Department of Surgery, Yokohama City University, 9Department of Surgery, Niigata University Graduate School of Medical and Dental Sciences, 10Department of Surgery, Fukushima Medical University

We introduce a murine orthotopic breast cancer model and radical mastectomy model with bioluminescence technology to quantify the tumor burden to mimic human breast cancer progression.

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Neuroscience

Quantitative Cell Biology of Neurodegeneration in Drosophila Through Unbiased Analysis of Fluorescently Tagged Proteins Using ImageJ
Jennifer M. Brazill 1, Yi Zhu 1, Chong Li 1, R. Grace Zhai 1,2
1Department of Molecular and Cellular Pharmacology, University of Miami Miller School of Medicine, 2School of Pharmacy, Key Laboratory of Molecular Pharmacology and Drug Evaluation (Yantai University), Ministry of Education, Collaborative Innovation Center of Advanced Drug Delivery System and Biotech Drugs in Universities of Shandong, Yantai University

We have developed a simple and adaptable workflow to extract quantitative data from fluorescence-imaging-based cell biological studies of protein aggregation and autophagic flux in the central nervous system of Drosophila models of neurodegeneration.

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Neuroscience

Purification of H3 and H4 Histone Proteins and the Quantification of Acetylated Histone Marks in Cells and Brain Tissue
Karolina J. Janczura 1,2, Claude-Henry Volmar 1,2, Claes Wahlestedt 1,2
1Department of Psychiatry & Behavioral Sciences, University of Miami Miller School of Medicine, 2Center for Therapeutic Innovation, University of Miami Miller School of Medicine

The purpose of this article is to provide a comprehensive, systematic guide to the efficient purification of histones H3 and H4 and the quantification of acetylated histone residues.

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Immunology and Infection

Using a Bacterial Pathogen to Probe for Cellular and Organismic-level Host Responses
Petoria Gayle 1, Nancy E. Freitag 2, Natasa Strbo 1, Kurt Schesser 1
1Department of Microbiology & Immunology, University of Miami Miller School of Medicine, 2Department of Microbiology & Immunology, University of Illinois College of Medicine

We describe both in vitro and in vivo infection assays that can be used to analyze the activities of host-encoding factors.

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Immunology and Infection

Isolation of Lamina Propria Mononuclear Cells from Murine Colon Using Collagenase E
Duneia McManus 1,2,3,4, Horacio J. Novaira 1,2,3,4, Anouk A.J. Hamers 1,2,3,4, Asha B. Pillai 1,2,3,4,5
1Department of Pediatrics, Division of Hematology / Oncology and Bone Marrow Transplantation, University of Miami Miller School of Medicine, 2Batchelor Children's Research Institute, University of Miami Miller School of Medicine, 3Department of Microbiology & Immunology, University of Miami Miller School of Medicine, 4Sylvester Comprehensive Cancer Center, University of Miami Miller School of Medicine, 5Holtz Children's Hospital, University of Miami Miller School of Medicine

The goal of this protocol is to isolate mononuclear cells that reside in the lamina propria of the colon by enzymatic digestion of the tissue using collagenase. This protocol allows for the efficient isolation of mononuclear cells resulting in a single cell suspension which in turn can be used for robust immunophenotyping.

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Medicine

A Computerized Functional Skills Assessment and Training Program Targeting Technology Based Everyday Functional Skills
Philip D. Harvey 1,2, Lize Tibiriçá 2,3, Peter Kallestrup 2, Sara J. Czaja 1,2,4
1University of Miami Miller School of Medicine, 2i-Function, 3Albizu University, 4Weill-Cornell Medical Center

This training protocol uses computerized training to teach technology-related everyday functional skills. These skills include financial skills, travel and transit, as well as medication management.

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Developmental Biology

A Neurite Outgrowth Assay and Neurotoxicity Assessment with Human Neural Progenitor Cell-Derived Neurons
Amir Bagheri 1,2, Seyedeh Fatemeh Razavipour 3, Claes Wahlestedt 2, Seyed Javad Mowla 1, Mohammad Ali Faghihi 2,4
1Department of Molecular Genetics, Faculty of Biological Sciences, Tarbiat Modares University, 2Center for Therapeutic Innovation and Department of Psychiatry & Behavioral Sciences, University of Miami Miller School of Medicine, 3Department of Biochemistry and Molecular Biology, University of Miami Miller School of Medicine, 4Persian BayanGene Research and Training Center

The presented protocol describes a method for a neurite outgrowth assay and neurotoxicity assessment of small molecule compounds.

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Bioengineering

Detergent-Free Decellularization of the Human Pancreas for Soluble Extracellular Matrix (ECM) Production
Riccardo Tamburrini 1,2,3, Deborah Chaimov 1,3, Amish Asthana 1,3, Carlo Gazia 3,4, Kevin Enck 3, Sean M. Muir 5, Justine Mariam Aziz 6, Sandrine Lablanche 7, Emily Tubbs 7, Alice A. Tomei 8,9, Mark Van Dyke 10, Shay Soker 3, Emmanuel C. Opara 3, Giuseppe Orlando 1,3
1Department of Surgery, Wake Forest Baptist Medical Center, 2Department of General Surgery, PhD Program in Experimental Medicine, University of Pavia, 3Wake Forest Institute for Regenerative Medicine, Wake Forest School of Medicine, 4Department of Surgery, Tor Vergata University of Rome, 5Wake Forest University College of Arts and Science, 6Wake Forest University School of Medicine, 7Laboratory of Fundamental and Applied Bioenergetics (LBFA), and Environmental and System Biology (BEeSy), Grenoble Alps University, 8Department of Biomedical Engineering, University of Miami, 9Diabetes Research Institute, University of Miami Miller School of Medicine, 10Department of Biomedical Engineering and Mechanics, School of Biomedical Engineering and Sciences, Virginia Polytechnic Institute and State University

The protocol described in this manuscript explains the steps for the fabrication of a soluble extracellular matrix (ECM) from the human pancreas. The solubilized ECM powder obtained through this protocol may be used for the recapitulation of pancreatic islets’ microenvironment in vitro and, potentially, in vivo settings.

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Medicine

Electroretinogram Recording for Infants and Children under Anesthesia to Achieve Optimal Dark Adaptation and International Standards
Byron L. Lam 1, Carlos Mendoza-Santiestaban 1, Alex Gonzalez 1,2, Cornelis Rowaan 1,2, Mu Liu 1, Joanne Martin 1, Steven Gayer 1,3, Osmany Gil Figueredo 1, Jean-Marie Parel 1,2
1Department of Ophthalmology, Bascom Palmer Eye Institute, University of Miami Miller School of Medicine, 2Ophthalmic Biophysics Center, Bascom Palmer Eye Institute, University of Miami Miller School of Medicine, 3Department of Anesthesiology, Bascom Palmer Eye Institute, University of Miami Miller School of Medicine

Adhering to international standards and maintaining retinal dark adaptation are critical to acquire valid full-field electroretinogram responses in the diagnosis and management of inherited retinal diseases. A practical protocol using a portable darkroom is provided to obtain full-field electroretinogram for infants and children under sedation or general anesthesia in the operating room setting.

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Medicine

High-Resolution Three-Dimensional Imaging of the Footpad Vasculature in a Murine Hindlimb Gangrene Model
Antoine J. Ribieras 1, Yulexi Y. Ortiz 1,2, Sophie Shrestha 3, C. Theodore Huerta 1, Hongwei Shao 1,2, Maria E. Boulina 4, Roberto I. Vazquez-Padron 1,2, Zhao-Jun Liu 1,2, Omaida C. Velazquez 1,2
1Dewitt Daughtry Family Department of Surgery, University of Miami Miller School of Medicine, 2Vascular Biology Institute, University of Miami Miller School of Medicine, 3University of Miami, 4Analytical Imaging Core Facility, University of Miami

The present protocol describes a unique, clinically relevant model of peripheral arterial disease that combines femoral artery and vein electrocoagulation with the administration of a nitric oxide synthase inhibitor to induce hindlimb gangrene in FVB mice. Intracardiac DiI perfusion is then used for high-resolution, three-dimensional imaging of the footpad vasculature.

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Cancer Research

Preparation of Cytoplasmic and Nuclear Long RNAs from Primary and Cultured Cells
Jacob Jahn 1, Sana Chaudhry 1, Maurizio Affer 1, Alejandro Pardo 1, Gabriel Pardo 1, Justin Taylor 1
1Sylvester Comprehensive Cancer Center, University of Miami Miller School of Medicine

The present protocol offers an efficient and flexible method to isolate RNA from nuclear and cytoplasmic fractions using cultured cells, and then validate using qPCR. This effectively serves as a replacement for other RNA preparation kits.

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Immunology and Infection

Comprehensive Approach for Microbial Isolation from Hidradenitis Suppurativa Tunnels
Tammy Gonzalez *1, Elina V. Zhivov *1, Raji R. Nagalla 1, Rebecca Verpile 1, Viviane Abreu Nunes 1, Marjana Tomic-Canic 1, Barry Resnik 1, Hadar Lev-Tov 1, Irena Pastar 1
1Dr. Phillip Frost Department of Dermatology and Cutaneous Surgery, University of Miami Miller School of Medicine

We showcase a method to successfully isolate fastidious and anaerobic organisms from cutaneous sinus tracts (tunnels) in tissues excised from patients with Hidradenitis Suppurativa.

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