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University at Albany, State University of New York

9 ARTICLES PUBLISHED IN JoVE

Medicine

A Protocol for Comprehensive Assessment of Bulbar Dysfunction in Amyotrophic Lateral Sclerosis (ALS)

Yana Yunusova^1,2, Jordan R. Green³, Jun Wang³, Gary Pattee⁴, Lorne Zinman^2,5

¹Department of Speech-Language Pathology, University of Toronto, ²ALS/ MN Clinic, Sunnybrook Health Science Centre, ³Department of Special Education and Communication Disorders, University of Nebraska-Lincoln, ⁴Department of Neurology, Munroe-Meyer Institute, University of Nebraska Medical Center, ⁵Department of Neurology, University of Toronto

Objective assessments of the physiological mechanisms that support speech are needed to monitor disease onset and progression in persons with ALS and to quantify treatment effects in clinical trials. In this video, we present a comprehensive, instrumentation-based protocol for quantifying speech motor performance in clinical populations.

Bioengineering

Nanomanipulation of Single RNA Molecules by Optical Tweezers

William Stephenson¹, Gorby Wan², Scott A. Tenenbaum^3,4, Pan T. X. Li^4,5

¹Nanoscale Engineering Graduate Program, College of Nanoscale Science and Engineering, University at Albany, State University of New York, ²Nanoscale Science Undergraduate Program, College of Nanoscale Science and Engineering, University at Albany, State University of New York, ³Nanobioscience Constellation, College of Nanoscale Science and Engineering, University at Albany, State University of New York, ⁴The RNA Institute, University at Albany, State University of New York, ⁵Department of Biological Sciences, University at Albany, State University of New York

Optical tweezers have been used to study RNA folding by stretching individual molecules from their 5’ and 3’ ends. Here common procedures are described to synthesize RNA molecules for tweezing, calibration of the instrument, and methods to manipulate single molecules.

Chemistry

Flow-pattern Guided Fabrication of High-density Barcode Antibody Microarray

Lisa S. Ramirez^1,2, Jun Wang^1,2

¹Department of Chemistry, University at Albany, State University of New York, ²Multiplex Biotechnology Laboratory, Cancer Research Center

This protocol outlines the fabrication of a large-scale, multiplexed two-dimensional DNA or antibody array, with potential applications in cell signaling studies and biomarker detection.

Medicine

Construction and Evaluation of a Murine Calvarial Osteolysis Model by Exposure to CoCrMo Particles in Aseptic Loosening

Hui Jiang^*1, Yicun Wang^*1, Zhantao Deng^*1,2,3, Jiewen Jin^2,3, Jia Meng¹, Shuo Chen¹, Jun Wang¹, Yang Qiu¹, Ting Guo¹, Jianning Zhao¹

¹Department of Orthopedics, Jinling Hospital, School of Medicine, Nanjing University, ²Center for Translational Medicine, Nanjing University Medical School, ³Jiangsu Key Laboratory for Molecular Medicine, Nanjing University Medical School

This manuscript describes a murine calvarial osteolysis model by exposure to CoCrMo particles, which constitutes an ideal animal model for assessing the interactions between wear particles and various cells in aseptic loosening.

Developmental Biology

A RANKL-based Osteoclast Culture Assay of Mouse Bone Marrow to Investigate the Role of mTORC1 in Osteoclast Formation

Qinggang Dai^*1, Yujiao Han^*2, Furong Xie^*1, Xuhui Ma³, Zhan Xu², Xiao Liu¹, Weiguo Zou², Jun Wang¹

¹Department of Pediatric Dentistry, Ninth People's Hospital, Shanghai Jiaotong University School of Medicine, Shanghai Key Laboratory of Stomatology & Shanghai Research Institute of Stomatology, National Clinical Research Center of Stomatology, ²State Key Laboratory of Cell Biology, CAS Center for Excellence in Molecular Cell Science, Shanghai Institute of Biochemistry and Cell Biology, University of Chinese Academy of Sciences, ³Department of Oral and Maxillofacial-Head and Neck Oncology, Ninth People's Hospital, Shanghai Jiaotong University School of Medicine

This manuscript describes a protocol to isolate and culture osteoclasts in vitro from mouse bone marrow, and to study the role of the mammalian/mechanistic target of rapamycin complex 1 in osteoclast formation.

Chemistry

Fabrication of Spherical and Worm-shaped Micellar Nanocrystals by Combining Electrospray, Self-assembly, and Solvent-based Structure Control

Xinyi Ding^1,2,3, Yuxiang Sun^1,2,3, Yanming Chen^1,2,3, Wanchuan Ding^1,2,3, Steven Emory^1,2,3,4, Tianhao Li^1,2,3, Zixing Xu^1,2,3, Ning Han^1,2,3, Jun Wang^1,2,3, Gang Ruan^1,2,3

¹Department of Biomedical Engineering, College of Engineering and Applied Sciences, Nanjing University, ²Institute of Materials Engineering, College of Engineering and Applied Sciences, Nanjing University, ³Collaborative Innovation Center of Chemistry for Life Sciences, Nanjing University, ⁴Department of Chemistry, Western Washington University

The present work describes a method to fabricate micellar nanocrystals, an emerging major class of nanobiomaterials. This method combines top-down electrospray, bottom-up self-assembly, and solvent-based structure control. The fabrication method is largely continuous, can produce high quality products, and possesses an inexpensive means of structure control.

Genetics

Culturing and Manipulation of O9-1 Neural Crest Cells

Bao H. Nguyen¹, Mamoru Ishii², Robert E. Maxson², Jun Wang¹

¹Molecular Physiology and Biophysics, Baylor College of Medicine, ²Department of Biochemistry and Molecular Biology, University of Southern California

O9-1 is a multipotent mouse neural crest cell line. Here we describe detailed step-by-step protocols for culturing O9-1 cells, differentiating O9-1 cells into specific cell types, and genetically manipulating O9-1 cells by using siRNA-mediated knockdown or CRISPR-Cas9 genome editing.

Behavior

Application of 3D Printing in the Construction of Burr Hole Ring for Deep Brain Stimulation Implants

Jiazhi Chen^*1,2, Xinyu Chen^*3, Siyuan Lv², Yuzhen Zhang¹, Hao Long¹, Kaijun Yang¹, Songtao Qi¹, Wangming Zhang², Jun Wang¹

¹Department of Neurosurgery, Nanfang Hospital, Southern Medical University, ²Department of Neurosurgery, Zhujiang Hospital, Southern Medical University, ³Faculty of Brain Sciences, University of College London

Here, we present a protocol to demonstrate 3D printing in the construction of deep brain stimulation implants.

Developmental Biology

An Optimized O9-1/Hydrogel System for Studying Mechanical Signals in Neural Crest Cells

Tram P. Le^*1, Xiaolei Zhao^*1, Shannon Erhardt^1,2, Jianhua Gu³, Huie Wang³, Tina O. Findley¹, Jun Wang^1,2

¹Department of Pediatrics, McGovern Medical School, The University of Texas Health Science Center at Houston, ²The University of Texas MD Anderson Cancer Center UTHealth Graduate School of Biomedical Sciences, The University of Texas Health Science Center at Houston, ³SEM AFM Core in the Houston Methodist Hospital Research Institute

Detailed step-by-step protocols are described here for studying mechanical signals in vitro using multipotent O9-1 neural crest cells and polyacrylamide hydrogels of varying stiffness.