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University of California, Riverside

14 ARTICLES PUBLISHED IN JoVE

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Biology

Amide Hydrogen/Deuterium Exchange & MALDI-TOF Mass Spectrometry Analysis of Pak2 Activation
Yuan-Hao Hsu 1, Jolinda A. Traugh 2
1Department of Chemistry, Tunghai University, 2Department of Biochemistry, University of California, Riverside

MALDI-TOF mass spectrometry was successfully utilized to monitor the amide hydrogen/deuterium exchange in protein kinase Pak2 activation.

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Immunology and Infection

High and Low Throughput Screens with Root-knot Nematodes Meloidogyne spp.
Hagop S. Atamian 1, Philip A. Roberts 1, Isgouhi Kaloshian 1
1Department of Nematology, University of California, Riverside

Two distinct methods to screen plants with root-knot nematodes are described. The described approaches include high-throughput screens with nematodes in a nondestructive manner facilitating the use of these plants in breeding programs.

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Immunology and Infection

Simple and Robust in vivo and in vitro Approach for Studying Virus Assembly
Sonali Chaturvedi 1, Bongsu Jung 2, Sharad Gupta 2, Bahman Anvari 2, A.L.N. Rao 1
1Department of Plant Pathology and Microbiology, University of California, Riverside , 2Department of Bioengineering, University of California, Riverside

A simple, efficient and robust way to synchronize the delivery of multiple viral components to plant cells via Agrobacterium-mediated transient expression is described. This approach is amenable for studying replication, encapsidation followed by in vitro reassembly of non-viral components into genome depleted optical viral ghosts suitable for biomedical applications.

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Medicine

Modeling Intracerebral Hemorrhage in Mice: Injection of Autologous Blood or Bacterial Collagenase
Paul R. Krafft 1, William B. Rolland 1, Kamil Duris 1, Tim Lekic 1, Aaron Campbell 2, Jiping Tang 1, John H. Zhang 1,3,4
1Department of Physiology and Pharmacology, Loma Linda University School of Medicine, 2College of Natural and Agricultural Sciences, University of California, Riverside , 3Department of Anesthesiology, Loma Linda University School of Medicine, 4Department of Neurosurgery, Loma Linda University School of Medicine

Clinically relevant animal models of intracerebral hemorrhage (ICH) are needed to extend our knowledge of hemorrhagic stroke and to examine novel therapeutic strategies. In this study, we describe and evaluate two ICH models that implement unilateral injections of either autologous whole blood or bacterial collagenase into the basal ganglia (corpus striatum) of mice.

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JoVE Journal

Quantitative FRET (Förster Resonance Energy Transfer) Analysis for SENP1 Protease Kinetics Determination
Yan Liu 1, Jiayu Liao 1
1Department of Bioengineering, Bourns College of Engineering, University of California, Riverside

A novel method involving quantitative analysis of FRET (Förster Resonance Energy Transfer) signals is described for studying enzyme kinetics. KM and kcat were obtained for the hydrolysis of the catalytic domain of SENP1 (SUMO/Sentrin specific protease 1) to pre-SUMO1 (Small Ubiquitin-like MOdifier). The general principles of this quantitative-FRET-based protease kinetic study can be applied to other proteases.

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Neuroscience

Thinned-skull Cortical Window Technique for In Vivo Optical Coherence Tomography Imaging
Jenny I. Szu 1, Melissa M. Eberle 2, Carissa L. Reynolds 2, Mike S. Hsu 1, Yan Wang 2, Christian M. Oh 2, M. Shahidul Islam 2, B. Hyle Park 2, Devin K. Binder 1
1Division of Biomedical Sciences, University of California, Riverside , 2Department of Bioengineering, University of California, Riverside

We present a method of creating a thinned-skull cortical window (TSCW) in a mouse model for in vivo OCT imaging of the cerebral cortex.

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External Magnetic Field Control of Intrinsic Electric Fields in Magneto-Electric Nanoparticles for Enabling Patient- and Disease-Specific Nanomedicine
Rakesh Guduru 1,2, Ping Liang 3, Jeongmin Hong 1,2, Madhavan Nair 1, Sakhrat Khizroev 1,2,3
1Center for Personalized Nanomedicine, Department of Immunology, Herbert Wertheim College of Medicine, Florida International University, 2Department of Electrical and Computer Engineering, Florida International University, 3Department of Electrical Engineering, University of California, Riverside

We describe the fabrication and characterization protocols used in our studies of magneto-electrical nanoparticles (MENs) for enabling Personalized Nanomedicine (PNM) from the perspective of fundamental physics and nanoengineering. This approach exploits quantum-mechanical coupling between electric and magnetic fields within MENs. Anti-Cancer and anti-HIV drugs were released on demand.

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Developmental Biology

Kidney Regeneration in Adult Zebrafish by Gentamicin Induced Injury
Caramai N. Kamei 1, Yan Liu 1,2, Iain A. Drummond 1,3
1Nephrology Division, Department of Medicine, Massachusetts General Hospital, 2Basic Sciences Division, Fred Hutchinson Cancer Research Center, 3Department of Genetics, Harvard Medical School

Here we present a reliable method to study adult kidney regeneration by inducing acute kidney injury by gentamicin injection. We show that injury is dependent on gentamicin dosage and environmental temperature using in situ hybridization to label lhx1a+ developing new nephrons.

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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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JoVE Journal

Isolation and Respiratory Measurements of Mitochondria from Arabidopsis thaliana
Wenhui Lyu 1, Jennifer Selinski 1, Lu Li 1, David A. Day 2, Monika W. Murcha 3, James Whelan 1, Yan Wang 1
1ARC Centre of Excellence in Plant Energy Biology, Department of Animal, Plant and Soil Science, School of Life Science, La Trobe University, 2School of Biological Sciences, Flinders University, 3ARC Centre of Excellence in Plant Energy Biology, University of Western Australia

As mitochondria are only a small percentage of the plant cell, they need to be purified for a range of studies. Mitochondria can be isolated from a variety of plant organs by homogenization, followed by differential and density gradient centrifugation to obtain a highly purified mitochondrial fraction.

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JoVE Journal

Chemoselective Preparation of 1-Iodoalkynes, 1,2-Diiodoalkenes, and 1,1,2-Triiodoalkenes Based on the Oxidative Iodination of Terminal Alkynes
Youzhi Li *1, Daya Huang *1, Ju Huang 1, Yan Liu 1, Keiji Maruoka 1,2
1School of Chemical Engineering and Light Industry, Guangdong University of Technology, 2Department of Chemistry, Graduate School of Science, Kyoto University

Herein, detailed protocols for the oxidative iodination of terminal alkynes using hypervalent-iodine reagents are presented, which chemoselectively afford 1-iodoalkynes, 1,2-diiodoalkenes, and 1,1,2-triiodoalkenes.

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Behavior

A Behavioral Test Battery for the Repeated Assessment of Motor Skills, Mood, and Cognition in Mice
Ran You 1,2,3, Yan Liu 3, Raymond Chuen-Chung Chang 3
1Nanjing Key Laboratory of Pediatrics, Children's Hospital of Nanjing Medical University, 2Nanjing Key Laboratory of Pediatrics, Nanjing Medical University, 3Laboratory of Neurodegenerative Disease, Li Ka Shing Faculty of Medicine, University of Hong Kong, Pokfulam, Hong Kong Special Administrative Region (HK SAR)

A comprehensive behavioral test battery of motor skills, mood—including social interaction, depression, and anxiety—and cognition is designed for the repeated assessment of neurodegeneration-related behavioral changes in mice.

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Medicine

Protocol and Guidelines for Point-of-Care Lung Ultrasound in Diagnosing Neonatal Pulmonary Diseases Based on International Expert Consensus
Jing Liu 1,2, Roberto Copetti 3, Erich Sorantin 4, Jovan Lovrenski 5, Javier Rodriguez-Fanjul 6, Dalibor Kurepa 7, Xing Feng 8, Luigi Cattaross 9, Huayan Zhang 10,11, Misun Hwang 12, Tsu F. Yeh 13,14, Yisrael Lipener 7, Abhay Lodha 15, Jia-Qin Wang 16, Hai-Ying Cao 2,17, Cai-Bao Hu 2,18, Guo-Rong Lyu 19, Xin-Ru Qiu 1,2, Li-Qun Jia 20, Xiao-Man Wang 20, Xiao-Ling Ren 1,2, Jiu-Ye Guo 1,2, Yue-Qiao Gao 1,2, Jian-Jun Li 1,2, Ying Liu 1,2, Wei Fu 1,2, Yan Wang 21, Zu-Lin Lu 1,2, Hua-Wei Wang 8, Li-Li Shang 22
1Department of Neonatology and NICU, Beijing Chaoyang District Maternal and Child Healthcare Hospital, 2The Neonatal Lung Ultrasound Training Base, Chinese College of Critical Ultrasound, 3Emergency Department, Cattinara University Hospital, 4Division of Pediatric Radiology, Department of Radiology, Medical University Graz, 5Faculty of Medicine, University of Novi Sad, Radiology Department, Institute for Children and Adolescents Health Care of Vojvodina, 6Pediatric Intensive Care Unit, Pediatric Service Hospital Joan XXIII Tarragona, University Rovira i Virgil, 7Division of Neonatal-Perinatal Medicine, Cohen Children's Medical Center, 8Department of Neonatology, Children's Hospital of Soochow University, 9Department of Neonatology, Udine University Hospital, 10Center for Newborn Care, Guangzhou Women and Children's Medical Center, 11Division of Neonatology, Children's Hospital of Philadelphia, 12Department of Radiology, Children's Hospital of Philadelphia, 13Division of Neonatology and NICU, Cook County Children's Hospital, University of Illinois, 14Division of Neonatology, Department of Pediatrics, Taipei Medical University, 15Department of Pediatrics and Community Health Sciences, University of Calgary, 16Department of Pediatrics, The Third Affiliated Hospital of Xinxiang Medical University, 17Department of Ultrasound, GE Healthcare, 18Intensive Care Unit, Zhejiang Hospital, 19Collaborative Innovation Center for Maternal and Infant Health Service Application Technology, Quanzhou Medical College, 20Department of Ultrasound, Beijing Children's Hospital Affiliated with Capital Medical University, 21Department of Neonatology and NICU, Tai'an City Central Hospital of Shandong Province, 22Department of Intensive Care Unit, The Second Affiliated Hospital of Heilongjiang University of Chinese Medicine

Lung ultrasound is a noninvasive and valuable tool for bedside evaluation of neonatal lung diseases. However, a relative lack of reference standards, protocols and guidelines may limit its application. Here, we aim to develop a standardized neonatal lung ultrasound diagnostic protocol to be used in clinical decision-making.

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Biology

Laser-Capture Microdissection RNA-Sequencing for Spatial and Temporal Tissue-Specific Gene Expression Analysis in Plants
Lim Chee Liew 1,2, Yan Wang 1,2, Marta Peirats-Llobet 1, Oliver Berkowitz 1,2,3, James Whelan 1,2,3, Mathew G. Lewsey 1,3
1Department of Animal, Plant and Soil Science, AgriBio Building, La Trobe University, 2Australian Research Council Centre of Excellence in Plant Energy Biology, La Trobe University, 3Australian Research Council Research Hub for Medicinal Agriculture, AgriBio Building, La Trobe University

Presented here is a protocol for laser-capture microdissection (LCM) of plant tissues. LCM is a microscopic technique for isolating areas of tissue in a contamination-free manner. The procedure includes tissue fixation, paraffin embedding, sectioning, LCM and RNA extraction. RNA is used in the downstream tissue-specific, temporally resolved analysis of transcriptomes.

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