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Massachusetts General Hospital, Harvard Medical School

41 ARTICLES PUBLISHED IN JoVE

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Biology

Monitoring Acupuncture Effects on Human Brain by fMRI
Kathleen K. S. Hui 1, Vitaly Napadow 1, Jing Liu 1, Ming Li 1, Ovidiu Marina 1,2, Erika E. Nixon 1, Joshua D. Claunch 1, Lauren LaCount 1, Tara Sporko 1, Kenneth K. Kwong 1
1Department of Radiology, Massachusetts General Hospital and Harvard Medical School, 2William Beaumont Hospital

FMRI and physiological monitoring is used to study the effects of Acupuncture on the central and peripheral nervous systems. Acupuncture mobilizes a limbic-paralimbic-neocortical network, with great overlap with the default mode network, to modulate neurological activity, possibly related to its autonomic effect in the peripheral nervous system.

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Biology

Bioluminescence Imaging of Heme Oxygenase-1 Upregulation in the Gua Sha Procedure
Kenneth K. Kwong 1,2, Lenuta Kloetzer 1,2,3,4, Kelvin K. Wong 5,6, Jia-Qian Ren 1,2, Braden Kuo 1,2,3,4, Yan Jiang 7, Y. Iris Chen 1,2, Suk-Tak Chan 1,2,8, Geoffrey S. Young 9, Stephen T.C. Wong 5,6
1Department of Radiology, Massachusetts General Hospital, Harvard Medical School, 2Athinoula A. Martinos Center for Biomedical Imaging, Massachusetts General Hospital, Harvard Medical School, 3Gastrointestinal Unit, Massachusetts General Hospital, Harvard Medical School, 4Department of Medicine, Massachusetts General Hospital, Harvard Medical School, 5Center for biotechnology and Informatics, The Methodist Hospital Research Institute, 6Department of Radiology, The Methodist Hospital, Weill Cornell Medical College, 7Bejing University of Chinese Medicine, 8Department of Health Technology and Informatics, The Hong Kong Polytechnic University, 9Department of Radiology, Brigham and Women's Hospital, Harvard Medical School

Gua Sha, traditional Chinese therapeutic skin scraping, causes subcutaneous microvascular blood extravasation. We report a protocol of bioluminescence imaging of HO-1-luciferase transgenic mice to demonstrate that Gua Sha upregulates heme oxygenase-1 (HO-1) in multiple organs.

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Neuroscience

Functional Mapping with Simultaneous MEG and EEG
Hesheng Liu 1, Naoaki Tanaka 1, Steven Stufflebeam 1, Seppo Ahlfors 1, Matti Hämäläinen 1
1Athinoula A. Martinos Center for Biomedical Imaging, MGH - Massachusetts General Hospital

We use magnetoencephalography (MEG) and electroencephalography (EEG) to map brain areas involved in the processing of simple sensory stimuli.

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Biology

PuraMatrix Encapsulation of Cancer Cells
Adnan O. Abu-Yousif 1, Imran Rizvi 1,2, Conor L. Evans 1, Jonathan P. Celli 1, Tayyaba Hasan 1,3
1Wellman Center for Photomedicine Massachusetts General Hospital, Harvard Medical School, 2Thayer School of Engineering, Dartmouth College, 3Department of Dermatology, Harvard Medical School

This video demonstrates how to encapsulate and culture cancer cells in PuraMatrix, a commercially available self assembling peptide gel.

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Neuroscience

Cerebral Blood Oxygenation Measurement Based on Oxygen-dependent Quenching of Phosphorescence
Sava Sakadžić 1, Emmanuel Roussakis 2, Mohammad A. Yaseen 1, Emiri T. Mandeville 3, Vivek J. Srinivasan 1, Ken Arai 3, Svetlana Ruvinskaya 1, Weicheng Wu 1, Anna Devor 1,4, Eng H. Lo 3, Sergei A. Vinogradov 2, David A. Boas 1
1Optics Division, Athinoula A. Martinos Center for Biomedical Imaging, Department of Radiology, Massachusetts General Hospital and Harvard Medical School, 2Department of Biochemistry and Biophysics, University of Pennsylvania, 3Neuroprotection Research Laboratory, Departments of Radiology and Neurology, Massachusetts General Hospital and Harvard Medical School, 4Departments of Neurosciences and Radiology, University of California

We present an experimental procedure for measuring the partial pressure of oxygen (pO2) in cerebral vasculature based on oxygen-dependent quenching of phosphorescence. Animal preparation and imaging procedures were outlined for both large field of view CCD-based imaging of pO2 in rats and 2-photon excitation based imaging of pO2 in mice.

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Neuroscience

Magnetic Resonance Spectroscopy of live Drosophila melanogaster using Magic Angle Spinning
Valeria Righi 1,2,3, Yiorgos Apidianakis 2,4, Laurence G. Rahme 2,4, A. Aria Tzika 1,2,3
1NMR Surgical Laboratory, Department of Surgery, Massachusetts General Hospital, Harvard Medical School, 2Shriners Burn Institute, 3Department of Radiology, Athinoula A. Martinos Center of Biomedical Imaging, Harvard Medical School, 4Molecular Surgery Laboratory, Department of Surgery, Massachusetts General Hospital, Harvard Medical School

This technique enables the use of high-resolution magic angle spinning proton MR spectroscopy (HRMAS 1H-MRS) for molecular characterization of live Drosophila melanogaster with a conventional 14.1 tesla spectrometer equipped with an HRMAS probe.

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Bioengineering

Decellularization and Recellularization of Whole Livers
Basak E. Uygun 1, Gavrielle Price 1, Nima Saeidi 1, Maria-Louisa Izamis 1, Tim Berendsen 1, Martin Yarmush 1, Korkut Uygun 1
1Center for Engineering in Medicine, Massachusetts General Hospital, Harvard Medical School, Shriners Hospitals for Children

Perfusion decellularization is a novel technique to produce whole liver scaffolds that retains the organ's extracellular matrix composition and microarchitecture. Herein, the method of preparing whole organ scaffolds using perfusion decellularization and subsequent repopulation with hepatocytes is described. Functional and transplantable liver grafts can be generated using this technique.

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Immunology and Infection

Competitive Homing Assays to Study Gut-tropic T Cell Migration
Eduardo J. Villablanca 1, J. Rodrigo Mora 1
1Gastrointestinal Unit, Massachusetts General Hospital, Harvard Medical School

Competitive homing experiments allow to directly assessing the migratory properties of two different cell populations in a single mouse. Here we illustrate this procedure by comparing the migration of ex vivo-generated gut-tropic versus non-gut tropic T cells.

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Immunology and Infection

Use of an Optical Trap for Study of Host-Pathogen Interactions for Dynamic Live Cell Imaging
Jenny M. Tam 1, Carlos E. Castro 2, Robert J. W. Heath 3, Michael K. Mansour 1, Michael L. Cardenas 1, Ramnik J. Xavier 3, Matthew J. Lang 4, Jatin M. Vyas 1
1Department of Medicine, Division of Infectious Diseases, Massachusetts General Hospital, Harvard Medical School, 2Department of Mechanical and Aerospace Engineering, The Ohio State University, 3Center for Computational and Integrative Biology, Massachusetts General Hospital, Harvard Medical School, 4Dept. of Chemical and Biomolecular Engineering, Vanderbilt University

A method is described to individually select, manipulate, and image live pathogens using an optical trap coupled to a spinning disk microscope. The optical trap provides spatial and temporal control of organisms and places them adjacent to host cells. Fluorescence microscopy captures dynamic intercellular interactions with minimal perturbation to cells.

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Bioengineering

Video-rate Scanning Confocal Microscopy and Microendoscopy
Alexander J. Nichols 1,2,3, Conor L. Evans 1,3
1Program in Biophysics, Harvard University , 2Division of Health Sciences and Technology, Harvard-MIT, 3Wellman Center for Photomedicine, Massachusetts General Hospital, Harvard Medical School

The complete construction of a custom, real-time confocal scanning imaging system is described. This system, which can be readily used for video-rate microscopy and microendoscopy, allows for an array of imaging geometries and applications not accessible using standard commercial confocal systems, at a fraction of the cost.

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Medicine

Non-invasive Optical Measurement of Cerebral Metabolism and Hemodynamics in Infants
Pei-Yi Lin 1, Nadege Roche-Labarbe 1,2, Mathieu Dehaes 3, Stefan Carp 1, Angela Fenoglio 3, Beniamino Barbieri 4, Katherine Hagan 1, P. Ellen Grant 3, Maria Angela Franceschini 1
1Athinoula A. Martinos Center for Biomedical Imaging, Massachusetts General Hospital, Harvard Medical School, 2Lab. PALM, Université de Caen Basse-Normandie, 3Fetal-Neonatal Neuroimaging and Developmental Science Center, Boston Children's Hospital, Harvard Medical School, 4ISS, INC.

We combined frequency-domain near-infrared spectroscopy measures of cerebral hemoglobin oxygenation with diffuse correlation spectroscopy measures of cerebral blood flow index to estimate an index of oxygen metabolism. We tested the utility of this measure as a bedside screening tool to evaluate the health and development of the newborn brain.

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Biology

Assessing Murine Resistance Artery Function Using Pressure Myography
Mohd Shahid 1, Emmanuel S. Buys 1
1Anesthesia Center for Critical Care Research, Department of Anesthesia, Critical Care, and Pain Medicine, Massachusetts General Hospital, Harvard Medical School

In pressure myography, an intact small segment of a vessel is mounted onto two small cannulas and pressurized to a suitable luminal pressure. Here, we describe the method to measure vasorelaxation response of the mouse 3rd order mesenteric arteries in c57 and sGCα1-/- mice using pressure myography.

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Neuroscience

RNAi-mediated Double Gene Knockdown and Gustatory Perception Measurement in Honey Bees (Apis mellifera)
Ying Wang 1, Nicholas Baker 1, Gro V. Amdam 1,2
1School of Life Sciences, Arizona State University , 2Department of Chemistry, Biotechnology, and Food Science, Norwegian University of Life Sciences

In this protocol, we describe two strategies that simultaneously suppress two genes (double gene knockdown) in honey bees. Then we present how to use the proboscis extension response (PER) assay to study the effect of double gene knockdown on honey bee gustatory perception.

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Medicine

Ex Situ Normothermic Machine Perfusion of Donor Livers
Negin Karimian 1,2, Alix P.M. Matton 1,2, Andrie C. Westerkamp 1,2, Laura C. Burlage 1,2, Sanna op den Dries 1,2, Henri G.D. Leuvenink 2, Ton Lisman 1,2, Korkut Uygun 3, James F. Markmann 4, Robert J. Porte 1,2
1Section of Hepato-Pancreato-Biliary Surgery and Liver Transplantation, University of Groningen, University Medical Center Groningen, 2Surgical Research Laboratory, Department of Surgery, University of Groningen, University Medical Center Groningen, 3Center of Engineering in Medicine/Surgical Services, Massachusetts General Hospital, Harvard Medical School, and Shriners Burns Hospital, 4Division of Transplantation, Department of Surgery, Massachusetts General Hospital, Harvard Medical School

Here we present a protocol describing oxygenated ex situ machine perfusion of donor liver grafts. This article contains a step by step protocol to procure and prepare the liver graft for machine perfusion, prepare the perfusion fluid, prime the perfusion machine and perform oxygenated normothermic machine perfusion of the liver graft.

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Medicine

Functional Human Liver Preservation and Recovery by Means of Subnormothermic Machine Perfusion
Bote G. Bruinsma *1, James H. Avruch *2, Pepijn D. Weeder 1, Gautham V. Sridharan 1, Basak E. Uygun 1, Negin G. Karimian 1, Robert J. Porte 3, James F. Markmann 2, Heidi Yeh 2, Korkut Uygun 1
1Center for Engineering in Medicine, Dept. of Surgery, Massachusetts General Hospital, Harvard Medical School, 2Transplant Center, Dept. of Surgery, Massachusetts General Hospital, Harvard Medical School, 3Section of Hepatobiliary Surgery and Liver Transplantation, Department of Surgery, University Medical Center Groningen, University of Groningen

We describe a method of ex vivo machine perfusion of human liver grafts at subnormothermic temperature (21 °C).

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Bioengineering

Layer-by-layer Collagen Deposition in Microfluidic Devices for Microtissue Stabilization
William J. McCarty 1,2, Ljupcho Prodanov 1,2, Shyam Sundhar Bale 1,2, Abhinav Bhushan 1,2, Rohit Jindal 1,2, Martin L. Yarmush 1,2, O. Berk Usta 1,2
1Center for Engineering in Medicine, Massachusetts General Hospital, Harvard Medical School, 2Shriners Hospitals for Children-Boston

The creation of functional microtissues within microfluidic devices requires the stabilization of cell phenotypes by adapting traditional cell culture techniques to the limited spatial dimensions in microdevices. Modification of collagen allows the layer-by-layer deposition of ultrathin collagen assemblies that can stabilize primary cells, such as hepatocytes, as microfluidic tissue models.

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Medicine

Calcification of Vascular Smooth Muscle Cells and Imaging of Aortic Calcification and Inflammation
Caitlin O'Rourke *1, Georgia Shelton *1,2, Joshua D. Hutcheson 3,4, Megan F. Burke 2, Trejeeve Martyn 1, Timothy E. Thayer 2, Hannah R. Shakartzi 1, Mary D. Buswell 1, Robert E. Tainsh 1, Binglan Yu 1,4, Aranya Bagchi 1,4, David K. Rhee 2,4, Connie Wu 1,2,4, Matthias Derwall 5, Emmanuel S. Buys 1,4, Paul B. Yu 3,4, Kenneth D. Bloch 1,2,4, Elena Aikawa 3,4, Donald B. Bloch 1,5,6, Rajeev Malhotra 2,4
1Anesthesia Center for Critical Care Research of the Department of Anesthesia, Critical Care, and Pain Medicine, Massachusetts General Hospital, 2Cardiovascular Research Center and Cardiology Division of the Department of Medicine, Massachusetts General Hospital, 3Cardiovascular Division, Brigham and Women's Hospital, 4Harvard Medical School, 5Department of Anesthesiology, Uniklinik RWTH Aachen, RWTH Aachen University, 6Center for Immunology and Inflammatory Diseases and the Division of Rheumatology, Allergy, and Immunology of the Department of Medicine, Massachusetts General Hospital

Vascular calcification is an important predictor of and contributor to human cardiovascular disease. This protocol describes methods for inducing calcification of cultured primary vascular smooth muscle cells and for quantifying calcification and macrophage burden in animal aortas using near-infrared fluorescence imaging.

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JoVE Core

Interictal High Frequency Oscillations Detected with Simultaneous Magnetoencephalography and Electroencephalography as Biomarker of Pediatric Epilepsy
Christos Papadelis 1, Eleonora Tamilia 1, Steven Stufflebeam 2, Patricia E. Grant 1, Joseph R. Madsen 3, Phillip L. Pearl 4, Naoaki Tanaka 2
1Fetal-Neonatal Neuroimaging and Developmental Science Center, Division of Newborn Medicine, Department of Medicine, Boston Children's Hospital, Harvard Medical School, 2Athinoula A. Martinos Center for Biomedical Imaging, Massachusetts General Hospital, Harvard Medical School, 3Division of Epilepsy Surgery, Department of Neurosurgery, Boston Children's Hospital, Harvard Medical School, 4Division of Epilepsy and Clinical Neurophysiology, Department of Neurology, Boston Children's Hospital, Harvard Medical School

High Frequency Oscillations (HFOs) have emerged as presurgical biomarkers for the identification of the epileptogenic zone in pediatric patients with medically refractory epilepsy. A methodology for the noninvasive recording, detection, and localization of HFOs with simultaneous scalp electroencephalography (EEG) and magnetoencephalography (MEG) is presented.

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Immunology and Infection

In Vivo Investigation of Antimicrobial Blue Light Therapy for Multidrug-resistant Acinetobacter baumannii Burn Infections Using Bioluminescence Imaging
Yucheng Wang 1,2,3, Olivia D. Harrington 1, Ying Wang 1, Clinton K. Murray 4, Michael R. Hamblin 1, Tianhong Dai 1
1Wellman Center for Photomedicine, Massachusetts General Hospital, Harvard Medical School, 2Department of Medical Oncology, Beijing Institute of Translational Medicine, Chinese Academy of Sciences, 3Cancer Center, Aviation General Hospital, Beijing, 4Infectious Disease Service, Brooke Army Medical Center

Infections caused by multidrug-resistant (MDR) bacterial strains have emerged as a serious threat to public health, necessitating the development of alternative therapeutics. We present a protocol to evaluate the effectiveness of antimicrobial blue light (aBL) therapy for MDR Acinetobacter baumannii infections in mouse burns by using bioluminescence imaging.

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Cancer Research

A Comprehensive Procedure to Evaluate the In Vitro Performance of the Putative Hemangioblastoma Neovascularization Using the Spheroid Sprouting Assay
Ying Wang 1, DanQi Chen 1, MingYu Chen 1, KaiYuan Ji 1, DeXuan Ma 1, LiangFu Zhou 1
1Department of Neurosurgery, Huashan Hospital, Fudan University

This paper presents a comprehensive procedure to evaluate in vitro whether classic tumor angiogenesis exists in hemangioblastomas (HBs) and its role in HBs. The results highlight the complexity of HB-neovascularization and suggest that this common form of angiogenesis is only a complementary mechanism in the HB-neovascularization.

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Neuroscience

Neurovascular Network Explorer 2.0: A Simple Tool for Exploring and Sharing a Database of Optogenetically-evoked Vasomotion in Mouse Cortex In Vivo
Hana Uhlirova 1,2, Peifang Tian 3,4, Kıvılcım Kılıç 3,5, Martin Thunemann 1, Vishnu B. Sridhar 6, Radim Chmelik 2,7, Hauke Bartsch 1, Anders M. Dale 1,3, Anna Devor 1,3,8, Payam A. Saisan 3
1Department of Radiology, University of California, San Diego, 2Central European Institute of Technology, Brno University of Technology, 3Department of Neurosciences, University of California, San Diego, 4Department of Physics, John Carroll University, 5Department of Biomedical Engineering, Boston University, 6Bioengineering Undergraduate Program, University of California, San Diego, 7Institute of Physical Engineering, Faculty of Mechanical Engineering, Brno University of Technology, 8Martinos Center for Biomedical Imaging, Massachusetts General Hospital, Harvard Medical School

A graphical user interface for exploring and sharing a database of optogenetically-induced vascular responses in mouse somatosensory cortex in vivo measured by 2-photon microscopy is presented. It allows browsing the data, criteria-based selection, averaging, localization of measurements within a 3D volume of vasculature and exporting the data.

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Engineering

Radio Frequency Magnetron Sputtering of GdBa2Cu3O7δ/ La0.67Sr0.33MnO3 Quasi-bilayer Films on SrTiO3 (STO) Single-crystal Substrates
Ying Wang 1,2, Zhen Li 1,2, YongSheng Liu 1, Yijie Li 2, Linfei Liu 2, Da Xu 2, Xiaojing Luo 1, Tian Gao 1,3, Yanyan Zhu 1,4, Luozeng Zhou 3, Jianming Xu 3
1Department of Physics, Mathematics, Shanghai Key Laboratory of Materials Protection and Advanced Materials in Electric Power, Shanghai University of Electric Power, 2Key Laboratory of Artificial Structure and Quantum Control, Ministry of Education, Department of Physics, Shanghai Jiao Tong University, 3Shanghai Institute of Space Power-sources, 4Shanghai Key Laboratory of High Temperature Superconductors, Shanghai University

Here, we present a protocol to grow LSMO nanoparticles and (Gd) BCO films on (001) SrTiO3 (STO) single-crystal substrates by radio frequency (RF)-sputtering.

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Bioengineering

Mouse Model of Pressure Ulcers After Spinal Cord Injury
Suneel Kumar 1, Yuying Tan 1, Martin L. Yarmush 1,2, Biraja C. Dash 3, Henry C. Hsia 3, Francois Berthiaume 1
1Department of Biomedical Engineering, Rutgers University, 2Center for Engineering in Medicine, Massachusetts General Hospital, Harvard Medical School, Shriners Hospitals for Children, 3Department of Surgery, Yale School of Medicine, Yale University

Here, we describe a simple method to induce clinically relevant skin pressure ulcers (PUs) in a mouse model of spinal cord injury (SCI). This model can be used in pre-clinical studies to screen for different therapeutics for healing PUs in SCI patients.

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Immunology and Infection

Human Serum Anti-aquaporin-4 Immunoglobulin G Detection by Cell-based Assay
Caiyun Liu 1, Mingqin Zhu 1, Ying Wang 1
1Department of Neurology and Neuroscience Center, The First Hospital of Jilin University

Cell-based assay is a widely used method to detect serum anti-aquaporin-4 immunoglobulin G. This method could be applied to clinical diagnosis and scientific researches of neuromyelitis optical spectrum disorders.

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JoVE Core

Functional MRI in Conjunction with a Novel MRI-compatible Hand-induced Robotic Device to Evaluate Rehabilitation of Individuals Recovering from Hand Grip Deficits
Mark P. Ottensmeyer 1,2, Shasha Li 2,3,4, Gianluca De Novi 1,2, A. Aria Tzika 2,3,4
1Medical Device & Simulation Laboratory, Department of Radiology, Massachusetts General Hospital, 2Harvard Medical School, 3NMR Surgical Laboratory, Department of Surgery, Center for Surgery, Innovation and Bioengineering, Massachusetts General Hospital, Harvard Medical School, 4Athinoula A. Martinos Center of Biomedical Imaging, Department of Radiology, Massachusetts General Hospital, Harvard Medical School

We performed functional MRI using a novel MRI-compatible hand-induced robotic device to evaluate its utility for monitoring hand motor function in individuals recovering from neurological deficits.

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Genetics

A Bioinformatics Pipeline to Accurately and Efficiently Analyze the MicroRNA Transcriptomes in Plants
Ying Wang *1,2, Zheng Kuang *1,2, Lei Li 2, Xiaozeng Yang 1
1Beijing Key Laboratory of Agricultural Genetic Resources and Biotechnology, Beijing Agro-Biotechnology Research Center, Beijing Academy of Agriculture and Forestry Sciences, 2State Key Laboratory of Protein and Plant Gene Research, Peking-Tsinghua Center for Life Sciences, School of Advanced Agricultural Sciences and School of Life Sciences, Peking University

A bioinformatics pipeline, namely miRDeep-P2 (miRDP2 for short), with updated plant miRNA criteria and an overhauled algorithm, could accurately and efficiently analyze microRNA transcriptomes in plants, especially for species with complex and large genomes.

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Immunology and Infection

Lentiviral CRISPR/Cas9-Mediated Genome Editing for the Study of Hematopoietic Cells in Disease Models
Soichi Sano 1, Ying Wang 1, Megan A. Evans 1, Yoshimitsu Yura 1, Miho Sano 1, Hayato Ogawa 1, Keita Horitani 1, Heather Doviak 1, Kenneth Walsh 1
1Hematovascular Biology Center, Robert M. Berne Cardiovascular Research Center, University of Virginia School of Medicine

Described are protocols for the highly efficient genome editing of murine hematopoietic stem and progenitor cells (HSPC) by the CRISPR/Cas9 system to rapidly develop mouse model systems with hematopoietic system-specific gene modifications.

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Bioengineering

Bulk Droplet Vitrification for Primary Hepatocyte Preservation
Reinier J. de Vries 1,2,3, Peony D. Banik 1,2, Sonal Nagpal 1,2, Lindong Weng 1,2, Sinan Ozer 1,2, Thomas M. van Gulik 3, Mehmet Toner 1,2, Shannon N. Tessier 1,2, Korkut Uygun 1,2
1Center for Engineering in Medicine, Harvard Medical School, Massachusetts General Hospital, 2Shriners Hospitals for Children, Boston, 3Department of Surgery, University of Amsterdam

This manuscript describes an ice-free cryopreservation method for large quantities of rat hepatocytes whereby primary cells are pre-incubated with cryoprotective agents at a low concentration and vitrified in large droplets.

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JoVE Core

Bone Marrow Transplantation Procedures in Mice to Study Clonal Hematopoiesis
Eunbee Park 1, Megan A. Evans 4, Heather Doviak 4, Keita Horitani 4, Hayato Ogawa 4, Yoshimitsu Yura 4, Ying Wang 2, Soichi Sano 3,4, Kenneth Walsh 1,4
1Department of Biochemistry and Molecular Genetics, University of Virginia School of Medicine, 2Department of Cardiology, Xinqiao Hospital, Army Medical University, 3Department of Cardiology, Osaka City University Graduate School of Medicine, 4Hematovascular Biology Center, Robert M. Berne Cardiovascular Research Center, University of Virginia School of Medicine

We describe three methods of bone marrow transplantation (BMT): BMT with total-body irradiation, BMT with shielded irradiation, and BMT method with no pre-conditioning (adoptive BMT) for the study of clonal hematopoiesis in mouse models.

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Immunology and Infection

Partial Heterotopic Hindlimb Transplantation Model in Rats
Marion Goutard 1,2,3, Mark A. Randolph 1,2,3, Corentin B. Taveau 1,2,3,4, Elise Lupon 1,2,3, Laurent Lantieri 4, Korkut Uygun 1,2,3, Curtis L. Cetrulo Jr. *1,2,3, Alexandre G. Lellouch *1,2,3,4
1Division of Plastic and Reconstructive Surgery, Massachusetts General Hospital, Harvard Medical School, 2Vascularized Composite Allotransplantation Laboratory, Center for Transplantation Sciences, Massachusetts General Hospital, Harvard Medical School, 3Shriners Hospital for Children, 4Service de Chirurgie Plastique, Hôpital Européen Georges Pompidou, Assistance Publique-Hôpitaux de Paris (APHP), Université de Paris

This paper presents a partial heterotopic osteomyocutaneous flap transplantation protocol in rats and its potential outcomes in the mid-term follow-up.

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Medicine

Visualizing and Quantifying Pharmaceutical Compounds within Skin using Coherent Raman Scattering Imaging
Benjamin A. Kuzma 1, Isaac J. Pence 1, Alexander Ho 1, Conor L. Evans 1
1Wellman Center for Photomedicine, Massachusetts General Hospital, Harvard Medical School

A coherent Raman scattering imaging methodology to visualize and quantify pharmaceutical compounds within the skin is described. This paper describes skin tissue preparation (human and mouse) and topical formulation application, image acquisition to quantify spatiotemporal concentration profiles, and preliminary pharmacokinetic analysis to assess topical drug delivery.

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Bioengineering

A Reliable Porcine Fascio-Cutaneous Flap Model for Vascularized Composite Allografts Bioengineering Studies
Victor Pozzo 1,2,4, Golda Romano 1,2,4, Marion Goutard 1,2,4, Elise Lupon 1,2,4, Pierre Tawa 1,2,4, Aylin Acun 3,4,5, Alec R. Andrews 2, Corentin B. Taveau 1,2,4,6, Basak E. Uygun 1,2,3,4, Mark A. Randolph 1,2,4, Curtis L. Cetrulo 1,2,4, Alexandre G. Lellouch 1,2,4,6
1Division of Plastic and Reconstructive Surgery, Massachusetts General Hospital, Harvard Medical School, 2Vascularized Composite Allotransplantation Laboratory, Center for Transplantation Sciences, Massachusetts General Hospital, Harvard Medical School, 3Center for Engineering in Medicine and Surgery, Massachusetts General Hospital, Harvard Medical School, 4Shriners Hospital for Children, 5Department of Biomedical Engineering, Widener University, 6Service de Chirurgie Plastique, Hôpital Européen Georges Pompidou, Assistance Publique-Hôpitaux de Paris (APHP), Université Paris Descartes

The present protocol describes the porcine fascio-cutaneous flap model and its potential use in vascularized composite tissue research.

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Bioengineering

Fabricating Highly Open Porous Microspheres (HOPMs) via Microfluidic Technology
Sheng-Chang Luo 1,2, Ying Wang 3, Ranjith Kumar Kankala 1,2, Yu Shrike Zhang 4, Ai-Zheng Chen 1,2
1Institute of Biomaterials and Tissue Engineering, Huaqiao University, 2Fujian Provincial Key Laboratory of Biochemical Technology, Huaqiao University, 3Affiliated Dongguan Hospital, Southern Medical University, 4Division of Engineering in Medicine, Department of Medicine, Brigham and Women's Hospital, Harvard Medical School

The present protocol describes the fabrication of poly(lactic-co-glycolic acid)-based highly open porous microspheres (HOPMs) via the single-emulsion formulation based facile microfluidic technology. These microspheres have potential applications in tissue engineering and drug screening.

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Neuroscience

Using a Bipolar Electrode to Create a Temporal Lobe Epilepsy Mouse Model by Electrical Kindling of the Amygdala
Yongchang Lu *1,2, Yang Dai *1,2, Siqi Ou 1,2, Yujing Miao 1, Ying Wang 1,2, Quanlei Liu 1,2, Yihe Wang 1,2, Penghu Wei 1,2, Yongzhi Shan 1,2, Guoguang Zhao 1,2,3
1Department of Neurosurgery, Xuanwu Hospital Capital Medical University, 2China Clinical Research Center for Epilepsy Capital Medical University, 3China Beijing Municipal Geriatric Medical Research Center

The amygdala plays a key role in temporal lobe epilepsy, which originates in and propagates from this structure. This article provides a detailed description of the fabrication of deep brain electrodes with both recording and stimulating functions. It introduces a model of medial temporal lobe epilepsy originating from the amygdala.

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Developmental Biology

Spatial High-resolution Analysis of Gene Expression Levels in Tendons
Steffany Villaseñor 1, Mor Grinstein 1,2,3
1Center for Regenerative Medicine, Department of Orthopedic Surgery, Massachusetts General Hospital, Harvard Medical School, 2Harvard Stem Cell Institute, 3Center Of Regeneration and Longevity (CORAL), Tel-Aviv Sourasky Medical Center

This article describes how to perform an optimized in situ protocol for tendons. This method discusses tissue preparation, section permeabilization, probe design, and signal amplification methods.

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Developmental Biology

Two-Photon Microscopy for the Study of Tendons
Steffany Villaseñor 1, Mor Grinstein 1,2,3
1Center for Regenerative Medicine, Department of Orthopedic Surgery, Massachusetts General Hospital, Harvard Medical School, 2Harvard Stem Cell Institute, 3Center Of Regeneration and Longevity (CORAL), Tel-Aviv Sourasky Medical Center

This article outlines the process of preparing, setting up, and imaging tendons using multiphoton microscopy. Additionally, it covers the application of SHG for analyzing collagen fibril alignment and the creation of a 3D representation of tendons. This methodology proves highly valuable in characterizing tendon cells and their ECM during injury and development.

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Neuroscience

Using Home-based, Remotely Supervised, Transcranial Direct Current Stimulation for Phantom Limb Pain
Kevin Pacheco-Barrios *1,2, Daniela Martinez-Magallanes *1, Cristina Xicota Naqui 1,3, Marianna Daibes 1, Elly Pichardo 1, Alejandra Cardenas-Rojas 1, David Crandell 4, Anahita Dua 5, Abhishek Datta 6,7, Wolnei Caumo 8,9,10, Felipe Fregni 1
1Neuromodulation Center and Center for Clinical Research Learning, Spaulding Rehabilitation Hospital, Massachusetts General Hospital, Harvard Medical School, 2Vicerrectorado de Investigación, Unidad de Investigación para la Generación y Síntesis de Evidencias en Salud, Universidad San Ignacio de Loyola, 3Nursing Department, Universitat Internacional de Catalunya, 4Spaulding Rehabilitation Hospital, Harvard Medical School, 5Division of Vascular and Endovascular Surgery, Massachusetts General Hospital, Harvard Medical School, 6Research and Development, Soterix Medical, 7City College of New York, 8Post-Graduate Program in Medical Sciences, School of Medicine, Universidade Federal do Rio Grande do Sul (UFRGS), 9Laboratory of Pain & Neuromodulation, Hospital de Clinicas de Porto Alegre (HCPA), 10Pain and Palliative Care Service, Hospital de Clinicas de Porto Alegre (HCPA)

The goal of this study is to describe a protocol for the home-based delivery of remotely supervised transcranial direct current stimulation (RS-tDCS) conserving the standard procedures of in-clinic practice, including safety, reproducibility, and tolerability. The participants included will be patients with phantom limb pain (PLP).

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Medicine

A Swine Burn Model for Investigating the Healing Process in Multiple Depth Burn Wounds
Yanis Berkane 1,2,3,4, Rohil Jain 2,5, Emmanuella O. Ajenu *2,5, Austin A. Shamlou *1,2, Khanh Nguyen 2,5, Michelle McCarthy 1,2,5, Basak E. Uygun 2,5, Alexandre G. Lellouch 1,2,6, Curtis L. Cetrulo Jr. 1,2, Korkut Uygun 2,5, Mark A. Randolph 1,2, Shannon N. Tessier 2,5
1Vascularized Composite Allotransplantation Laboratory, Massachusetts General Hospital, Harvard Medical School, 2Shriners Children’s Boston, 3Department of Plastic, Reconstructive and Aesthetic Surgery, Rennes University, 4SITI Laboratory, UMR, INSERM, Université de Rennes, 5Center for Engineering in Medicine and Surgery, Department of Surgery, Massachusetts General Hospital, Harvard Medical School, 6Innovative Therapies in Haemostasis, INSERM UMR-S, University of Paris

This protocol describes a reproducible multi-depth burn wound model in a Yucatan minipigs.

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Medicine

Cardiac Loading using Passive Left Atrial Pressurization and Passive Afterload for Graft Assessment
George Olverson IV *1, Manuela Lopera Higuita *2, Maya Bolger-Chen 2, Emmanuella O. Ajenu 2, Selena S. Li 1, Hussein Kharroubi 1, Bassel Tfayli 1, Chijioke Chukwudi 1, Nathan Minie 1, Joseph Catricala 1, Allison Pitti 1, William Michaud 1, Doug Vincent 3, David D’Alessandro 1, S. Alireza Rabi 1, Shannon N. Tessier 2, Asishana A. Osho 1
1Division of Cardiac Surgery, Massachusetts General Hospital, Harvard Medical School, 2Center for Engineering in Medicine and Surgery, Massachusetts General Hospital and Harvard Medical School, Shriners Children's Boston, 3VentriFlo Inc

The protocol describes a porcine ex vivo heart perfusion system in which direct loading of the left ventricle may serve as an assessment technique for graft health while simultaneously providing a holistic evaluation of graft function. A discussion of the system design and possible assessment metrics is also provided.

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Medicine

Monitoring Lung Function with Electrical Impedance Tomography in the Intensive Care Unit
Glasiele C. Alcala *1, Gamonmas Ekkapat *1,2, Kyle J. Medeiros 1, Caio C. A Morais 1, Yi Xin 1, Valentina Giammatteo 1,3, Giovanni Bruno 1, Alice Nova 1,4, Hatus Wanderley 1, Talisa Bühl 1, Marcus Victor 1,6, Timothy G. Gaulton 1, Carolyn J. La Vita 5, Marcelo B. P. Amato 7, Maurizio Ceradda 1, Lorenzo Berra 1, Roberta Ribeiro De Santis Santiago 1
1Department of Anesthesia, Critical Care and Pain Medicine, Massachusetts General Hospital, Harvard Medical School, 2Department of Anesthesiology, Faculty of Medicine, Chulalongkorn University. King Chulalongkorn Memorial Hospital, 3Department of Anesthesiology and Intensive Care Medicine, Catholic University of Sacred Heart, 4School of Medicine and Surgery, University of Milano-Bicocca, 5Department of Respiratory Care, Massachusetts General Hospital, Harvard Medical School, 6Electronics Engineering, Aeronautics Institute of Technology, 7Instituto do Coracao (InCor), Hospital das Clinicas HCFMUSP, Faculdade de Medicina, Universidade de Sao Paulo

Electrical Impedance Tomography is a non-invasive, radiation-free, real-time pulmonary ventilation monitoring tool. By measuring impedance changes in the thorax, it can visualize the distribution of air on a breath-by-breath basis. Initially intended for ventilation monitoring, electrical impedance tomography can also measure perfusion via intravenous injection of a saline solution.

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Medicine

Establishing a Swine Model to Study Uterus Dynamic Preservation and Transplantation
Lucile Cabanel 1,2,3, Haïzam Oubari 1,2,3, Ludivine Dion 4, Vincent Lavoué 4, Mark A. Randolph 2,3, Curtis L. Cetrulo 2,3,5, Korkut Uygun 1,2, Alexandre G. Lellouch 1,2,3,5, Yanis Berkane 1,2,6,7, Basak E. Uygun 1,2
1Center for Engineering in Medicine and Surgery, Massachusetts General Hospital, Harvard Medical School, 2Shriners Children's Boston, 3Plastic Surgery Research Laboratory, Department of plastic and reconstructive surgery, Massachusetts General Hospital, Harvard Medical School, 4Department of Gynecology and Obstetrics, CHU de Rennes, University of Rennes, 5Division of Plastic Surgery Research, Vascularized Composite Allotransplantation Program Development, Cedars-Sinai Hospital, 6Department of Plastic, Reconstructive and Aesthetic Surgery, CHU de Rennes, University of Rennes, 7SITI Laboratory, UMR1236 INSERM, Établissement Français du Sang, University of Rennes

A detailed and reproducible swine uterus model is described, from surgical procurement to the initiation of machine perfusion, allowing for the study of uterus preservation in transplantation.

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