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14 articles published in JoVE
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SILAC Based Proteomic Characterization of Exosomes from HIV-1 Infected Cells
Collins Cheruiyot1, Zemplen Pataki1, Robert Williams1, Bharat Ramratnam2,3, Ming Li3
1Brown University, 2COBRE Center for Cancer Research, Lifespan Laboratories, Rhode Island and Miriam Hospitals, 3Division of Infectious Diseases, Department of Medicine, Warren Alpert Medical School, Brown University
Here, we describe a quantitative proteomics method using the technique of stable isotope labeling by amino acids in cell culture (SILAC) to analyze the effects of HIV-1 infection on host exosomal proteomes. This protocol can be easily adapted to cells under different stress or infection conditions.
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Assessment and Evaluation of the High Risk Neonate: The NICU Network Neurobehavioral Scale
Barry M. Lester1,2, Lynne Andreozzi-Fontaine1,2, Edward Tronick3, Rosemarie Bigsby1,2
1Center for the Study of Children at Risk, Alpert Medical School, Brown University, 2Women & Infants Hospital of Rhode Island, 3University of Massachusetts, Boston
The NICU Network Neurobehavioral Scale (NNNS) was developed as an assessment for the at-risk infant. The purpose of this article is to describe the NNNS, provide video examples of the NNNS procedures and discuss the ways in which the exam has been used.
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Design of a Biaxial Mechanical Loading Bioreactor for Tissue Engineering
Bahar Bilgen1,2, Danielle Chu3, Robert Stefani1, Roy K. Aaron1,2
1Department of Orthopaedics, The Warren Alpert Brown Medical School of Brown University and the Rhode Island Hospital, 2Center for Restorative and Regenerative Medicine, VA Medical Center, Providence, RI, 3University of Texas Southwestern Medical Center
We designed a novel mechanical loading bioreactor that can apply uniaxial or biaxial mechanical strain to a cartilage biocomposite prior to transplantation into an articular cartilage defect.
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In Vivo Two-photon Imaging Of Experience-dependent Molecular Changes In Cortical Neurons
Vania Y. Cao1,2, Yizhou Ye1, Surjeet S. Mastwal1, David M. Lovinger3, Rui M. Costa4, Kuan H. Wang1
1Unit on Neural Circuits and Adaptive Behaviors, Genes Cognition and Psychosis Program, National Institute of Mental Health, 2Department of Neuroscience, Brown University - National Institutes of Health Graduate Partnership Program, 3Section on Synaptic Pharmacology, Laboratory for Integrative Neuroscience, National Institute on Alcohol Abuse and Alcoholism, 4Champalimaud Neuroscience Programme, Champalimaud Center for the Unknown
Experience-dependent molecular changes in neurons are essential for the brain's ability to adapt in response to behavioral challenges. An in vivo two-photon imaging method is described here that allows the tracking of such molecular changes in individual cortical neurons through genetically encoded reporters.
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Methods for Study of Neuronal Morphogenesis: Ex vivo RNAi Electroporation in Embryonic Murine Cerebral Cortex
Sofia B. Lizarraga1,2,3, Kathryn R. Coser1,2, Mark Sabbagh1,2, Eric M. Morrow1,2,3
1Department of Molecular, Cellular Biology and Biochemistry, Brown University, 2Institute for Brain Science, Brown University, 3Department of Psychiatry and Human Behavior, Warren Alpert School of Medicine, Brown University
To conduct a rapid assessment of the function of genes in the development of cerebral cortex, we describe methods involving the ex vivo electroporation of plasmids co-expressing inhibitory RNA (RNAi) and GFP in murine embryonic cortex. This protocol is amenable to the study of various aspects of neurodevelopment such as neurogenesis, neuronal migration and neuronal morphogenesis including dendrite and axon outgrowth.
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A Practical Approach to Genetic Inducible Fate Mapping: A Visual Guide to Mark and Track Cells In Vivo
Ashly Brown1, Stephen Brown2, Debra Ellisor2, Nellwyn Hagan1, Elizabeth Normand1, Mark Zervas2
1Department of Neuroscience, Division of Biology and Medicine, Brown University, 2Department of Molecular Biology, Cell Biology and Biochemistry, Division of Biology and Medicine, Brown University
Genetic Inducible Fate Mapping (GIFM) marks and tracks cells with fine spatial and temporal control in vivo and elucidates how cells from a specific genetic lineage contribute to developing and adult tissues. Demonstrated here are the techniques required to fate map E12.5 mouse embryos for epifluorescent and explant analysis.
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The MODS method for diagnosis of tuberculosis and multidrug resistant tuberculosis
Mark F Brady1, Jorge Coronel2, Robert H Gilman3, David AJ Moore4
1The Warren Alpert Medical School of Brown University, 2Laboratorio de Investigacion de Enfermedades Infecciosas, Universidad Peruana Cayetano Heredia, 3Department of International Health, Johns Hopkins Bloomberg School of Public Health, 4Wellcome Trust Centre for Clinical Tropical Medicine, Imperial College London
The microscopic-observation drug-susceptibility (MODS) assay is a low-cost, low-tech tool for high-performance detection of tuberculosis (TB) and multidrug-resistant tuberculosis (MDRTB). This video describes the MODS liquid media culture method.
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