University of North Dakota

8 ARTICLES PUBLISHED IN JoVE

Biology

Bioenergetic Profile Experiment using C2C12 Myoblast Cells

David G. Nicholls¹, Victor M. Darley-Usmar², Min Wu³, Per Bo Jensen³, George W. Rogers³, David A. Ferrick³

¹Buck Institute for Age Research, Novato, CA, ²Department of Pathology, Center for Free Radical Biology, University of Alabama at Birmingham - UAB, ³Seahorse Bioscience, North Billerica, MA

A description of a method for profiling mitochondrial function in cells is provided. The mitochondrial profile generated provides four parameters of mitochondrial function that can be measured in one experiment: basal respiration rate, ATP-linked respiration, proton leak, and reserve capacity.

Genetics

Semiconductor Sequencing for Preimplantation Genetic Testing for Aneuploidy

Baoheng Gui^*1,2,3, Yingxin Zhang^*4, Bo Liang⁵, Yvonne Ka Yin Kwok⁴, Wai Ting Lui⁴, Queenie Sum Yee Yeung⁴, Lingyin Kong⁶, Liming Xuan⁶, Jacqueline Pui Wah Chung⁴, Kwong Wai Choy^3,4

¹Department of Genetics and Metabolism, Maternal and Child Health Hospital of Guangxi Zhuang Autonomous Region, ²Birth Defects Prevention and Control Institute of Guangxi Zhuang Autonomous Region, ³Shenzhen Research Institute, The Chinese University of Hong Kong, ⁴Department of Obstetrics and Gynaecology, The Chinese University of Hong Kong, ⁵State Key Laboratory of Microbial Metabolism, Joint International Research Laboratory of Metabolic and Developmental Sciences, School of Life Sciences and Biotechnology, Shanghai Jiao Tong University, ⁶Basecare Medical Device Co., Ltd

Here, we introduce a semiconductor sequencing method for preimplantation genetic testing for aneuploidy (PGT-A) with the advantages of short turnaround time, low cost, and high throughput.

Immunology and Infection

Design of Cecal Ligation and Puncture and Intranasal Infection Dual Model of Sepsis-Induced Immunosuppression

Zhihan Wang^*1,2, Qinqin Pu^*1,2, Ping Lin^*2,3, Changlong Li¹, Jianxin Jiang³, Min Wu²

¹West China School of Basic Medical Sciences & Forensic Medicine, Sichuan University, ²Department of Biomedical Sciences, School of Medicine and Health Sciences, University of North Dakota, ³State Key Laboratory of Trauma, Burns and Combined Injury, Institute of Surgery Research, Daping Hospital, Army Medical University

This protocol describes techniques to measure infectious outcomes underlying secondary hospital-acquired infections in the immunosuppressive condition, first by establishing cecal ligation/puncture mice then challenging them with intranasal infection to create a clinically relevant model of immunosuppression sepsis.

JoVE Core

Mechanistic Insight into the Development of TNBS-Mediated Intestinal Fibrosis and Evaluating the Inhibitory Effects of Rapamycin

Ramkumar Mathur^1,2,3, Mahabub Maraj Alam⁴, Xiao-Feng Zhao⁴, Yunfei Huang⁴, Xinjun Zhu^1,2

¹Department of Molecular and Cellular Physiology, Albany Medical College, ²The IBD Center, Division of Gastroenterology, Department of Medicine, Albany Medical College, ³Department of Geriatrics, School of Medicine and Health Science, University of North Dakota, ⁴Department of Neuroscience and Experimental Therapeutics, Albany Medical College

In this study, we describe a detailed procedure of TNBS-mediated intestinal fibrosis, which exhibits comparable pathophysiology to Crohn's fibrosis. We also discuss this approach in light of rapamycin facilitated inhibitory effects on intestinal fibrosis.

Biochemistry

Generation and Assembly of Virus-Specific Nucleocapsids of the Respiratory Syncytial Virus

Yunrong Gao¹, Claire Ogilvie¹, Anirudh Raghavan¹, Chloe Von Hoffmann¹, Bo Liang¹

¹Department of Biochemistry, Emory University School of Medicine

For in-depth mechanistic analysis of the respiratory syncytial virus (RSV) RNA synthesis, we report a protocol of utilizing the chaperone phosphoprotein (P) for coexpression of the RNA-free nucleoprotein (N⁰) for subsequent in vitro assembly of the virus-specific nucleocapsids (NCs).

Neuroscience

Stereotaxic Viral Injection and Gradient-Index Lens Implantation for Deep Brain In Vivo Calcium Imaging

Rashmi Thapa¹, Bo Liang², Rongsong Liu³, Yun Li¹

¹Department of Zoology and Physiology, University of Wyoming, ²School of Electrical Engineering & Computer Science, University of North Dakota, ³Department of Mathematics and Statistics, University of Wyoming

Miniscope in vivo calcium imaging is a powerful technique to study neuronal dynamics and microcircuits in freely behaving mice. This protocol describes performing brain surgeries to achieve good in vivo calcium imaging using a miniscope.

JoVE Core

Atomic Force Microscopy Cantilever-Based Nanoindentation: Mechanical Property Measurements at the Nanoscale in Air and Fluid

Ashton E. Enrriques¹, Sean Howard², Raju Timsina³, Nawal K. Khadka³, Amber N. Hoover⁴, Allison E. Ray⁵, Ling Ding⁴, Chioma Onwumelu⁶, Stephan Nordeng⁶, Laxman Mainali^3,7, Gunes Uzer², Paul H. Davis^1,8

¹Micron School of Materials Science & Engineering, Boise State University, ²Department of Mechanical & Biomedical Engineering, Boise State University, ³Department of Physics, Boise State University, ⁴Energy and Environmental Science and Technology, Idaho National Laboratory, ⁵Science and Technology, Idaho National Laboratory, ⁶Harold Hamm School of Geology & Geological Engineering, University of North Dakota, ⁷Biomolecular Sciences Graduate Program, Boise State University, ⁸Center for Advanced Energy Studies

Quantifying the contact area and force applied by an atomic force microscope (AFM) probe tip to a sample surface enables nanoscale mechanical property determination. Best practices to implement AFM cantilever-based nanoindentation in air or fluid on soft and hard samples to measure elastic modulus or other nanomechanical properties are discussed.

Biology

Determining Ciliary Function and Membrane Impermeability of the Pseudostratified Lung Airway Epithelium

Muhammad Faheem¹, Masfique Mehedi¹

¹Department of Biomedical Sciences, School of Medicine and Health Sciences, University of North Dakota

Air-liquid interface culture is commonly used to develop pseudostratified airway epithelium by differentiating primary normal human bronchial epithelial cells that mimic the apical side of the lung airway. Here, we describe an easy protocol for determining its quality by monitoring its biophysical properties, such as ciliary function and membrane integrity.