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St. Jude Children’s Research Hospital

13 ARTICLES PUBLISHED IN JoVE

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Medicine

A Novel Rescue Technique for Difficult Intubation and Difficult Ventilation
Maria M. Zestos 1, Dima Daaboul 1, Zulfiqar Ahmed 1, Nasser Durgham 1, Roland Kaddoum 2
1Department of Anesthesia, Children’s Hospital of Michigan, 2Department of Anesthesia, St. Jude Children’s Research Hospital

We describe a technique to maintain oxygenation and ventilation using an endotracheal tube inserted nasally to the level of the naso-pharynx while sealing the mouth and nares for successful positive pressure ventilation.

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Neuroscience

Dissection and Imaging of Active Zones in the Drosophila Neuromuscular Junction
Rebecca Smith 1, J. Paul Taylor 1
1Developmental Neurobiology, St. Jude Children’s Research Hospital

The neuromuscular junction (NMJ) of Drosophila melanogaster is an important model system for studying normal synaptic function as well as perturbations to synaptic function found in certain neurological diseases. We present a protocol for dissection of the Drosophila larval motor system and immunostaining for active zone proteins within the NMJ.

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Biology

Modeling Asthma and Influenza Co-morbidity in C57BL/6J Mice
Amali Samarasinghe 1,2, Scott A Hoselton 3, Jane M Schuh 3, Jonathan A McCullers 1,2
1Department of Pediatrics, University of Tennessee Health Science Center, 2Department of Infectious Diseases, St. Jude Children’s Research Hospital, 3Department of Veterinary and Microbiological Sciences, North Dakota State University, Fargo

Asthma and influenza are diseases affecting the pulmonary system that affects millions worldwide. The aim of this study was to develop a mouse model of asthma and influenza comorbidity to study the intersection of these two diseases in the same host.

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Biochemistry

Deep Proteome Profiling by Isobaric Labeling, Extensive Liquid Chromatography, Mass Spectrometry, and Software-assisted Quantification
Anthony A. High 1, Haiyan Tan 1, Vishwajeeth R. Pagala 1, Mingming Niu 2,3, Ji-Hoon Cho 1, Xusheng Wang 1, Bing Bai 2, Junmin Peng 1,2
1St. Jude Proteomics Facility, St. Jude Children's Research Hospital, 2Departments of Structural Biology and Developmental Neurobiology, St. Jude Children's Research Hospital, 3Heilongjiang University of Chinese Medicine

We present a protocol to accurately quantitate proteins with isobaric labelling, extensive fractionation, bioinformatics tools, and quality control steps in combination with liquid chromatography interfaced to a high-resolution mass spectrometer.

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Cancer Research

Fabrication of Tongue Extracellular Matrix and Reconstitution of Tongue Squamous Cell Carcinoma In Vitro
Yupeng Yao 1, Weifan Lin 1, Yan Zhang 1
1Key Laboratory of Gene Engineering of the Ministry of Education, State Key Laboratory of Biocontrol, School of Life Sciences, Sun Yat-sen University

A method is shown here for the preparation of the tongue extracellular matrix (TEM) with efficient decellularization. The TEM can be used as functional scaffolds for the reconstruction of a tongue squamous cell carcinoma (TSCC) model under static or stirred culture conditions.

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Cancer Research

Using Microarrays to Interrogate Microenvironmental Impact on Cellular Phenotypes in Cancer
Rebecca Smith 1, Kaylyn Devlin 1, David Kilburn 1, Sean Gross 1, Damir Sudar 2, Elmar Bucher 1, Michel Nederlof 2, Mark Dane 1, Joe W. Gray 1, Laura Heiser 1, James E. Korkola 1
1Department of Biomedical Engineering, Knight Cancer Institute, Oregon Health and Science University, 2Quantitative Imaging Systems LLC

The purpose of the method presented here is to show how microenvironment microarrays (MEMA) can be fabricated and used to interrogate the impact of thousands of simple combinatorial microenvironments on the phenotype of cultured cells.

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Cancer Research

Three-Dimensional Bone Extracellular Matrix Model for Osteosarcoma
Yu Zhang 1, Yupeng Yao 1, Yan Zhang 1
1MOE Key Laboratory of Gene Function and Regulation, School of Life Sciences, Sun Yat-sen University

The bone extracellular matrix (BEM) model for osteosarcoma (OS) is well established and shown here. It can be used as a suitable scaffold for mimicking primary tumor growth in vitro and providing an ideal model for studying the histologic and cytogenic heterogeneity of OS.

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Chemistry

Cercosporin-Photocatalyzed [4+1]- and [4+2]-Annulations of Azoalkenes Under Mild Conditions
Philippe Icyishaka *1, Chenxing Li *1, Liushen Lu 1, Wenhao Bao 1, Jia Li 1, Yan Zhang 2, Yijian Rao 1
1Key Laboratory of Carbohydrate Chemistry and Biotechnology, Ministry of Education, School of Biotechnology, Jiangnan University, 2School of Pharmaceutical Science, Jiangnan University

New routes for the synthesis of nitrogen-containing heterocycles utilizing cercosporin as a metal-free photocatalyst were developed.

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Biochemistry

JUMPn: A Streamlined Application for Protein Co-Expression Clustering and Network Analysis in Proteomics
David Vanderwall 1, Poudel Suresh 1,2, Yingxue Fu 2, Ji-Hoon Cho 2, Timothy I. Shaw 2,3, Ashutosh Mishra 2, Anthony A. High 2, Junmin Peng 1,2, Yuxin Li 1,2
1Departments of Structural Biology and Developmental Neurobiology, St. Jude Children’s Research Hospital, 2Center for Proteomics and Metabolomics, St. Jude Children’s Research Hospital, 3Department of Computational Biology, St. Jude Children’s Research Hospital

We present a systems biology tool JUMPn to perform and visualize network analysis for quantitative proteomics data, with a detailed protocol including data pre-processing, co-expression clustering, pathway enrichment, and protein-protein interaction network analysis.

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Biology

Three-Dimensional, Serum-Free Culture System for Lacrimal Gland Stem Cells
Hongzi Chen 1, Pan Huang 1, Yan Zhang 1
1MOE Key Laboratory of Gene Function and Regulation, School of Life Sciences, Sun Yat-sen University

The three-dimensional, serum-free culture method for adult lacrimal gland (LG) stem cells is well established for the induction of LG organoid formation and differentiation into acinar or ductal-like cells.

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Developmental Biology

Fixation of Embryonic Mouse Tissue for Cytoneme Analysis
Eric T. Hall 1, Christina A. Daly 1,2, Yan Zhang 1, Miriam E. Dillard 1, Stacey K. Ogden 1
1Department of Cell and Molecular Biology, St. Jude Children’s Research Hospital, 2St. Jude Graduate School of Biomedical Sciences

Here, an optimized step-by-step protocol is provided for fixation, immunostaining, and sectioning of embryos to detect specialized signaling filopodia called cytonemes in developing mouse tissues.

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Biology

Protocols for CRISPR/Cas9 Mutagenesis of the Oriental Fruit Fly Bactrocera dorsalis
Jinxi Yuan 1, Jie Zhang 1, Yan Zhang 1, WuYun QiQiGe 1, Wei Liu 2, Shanchun Yan 1, Guirong Wang 2
1Key Laboratory of Sustainable Forest Ecosystem Management - Ministry of Education, Northeast Forestry University, 2Shenzhen Branch, Guangdong Laboratory of Lingnan Modern Agriculture, Genome Analysis Laboratory of the Ministry of Agriculture and Rural Affairs, Agricultural Genomics Institute at Shenzhen, Chinese Academy of Agricultural Sciences

This paper presents the step-by-step protocols for CRISPR/Cas9 mutagenesis of the Oriental fruit fly Bactrocera dorsalis. Detailed steps provided by this standardized protocol will serve as a useful guide for generating mutant flies for functional gene studies in B. dorsalis.

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Cancer Research

Measurement of the Compressibility of Cell and Nucleus Based on Acoustofluidic Microdevice
Qibin Fu 1, Yan Zhang 1, Tuchen Huang 1, Yang Liu 1
1Sino-French Institute of Nuclear Engineering and Technology, Sun Yat-sen University

Here a protocol is presented to build a fast and non-destructive system for measuring cell or nucleus compressibility based on acoustofluidic microdevice. Changes in mechanical properties of tumor cells after epithelial-mesenchymal transition or ionizing radiation were investigated, demonstrating the application prospect of this method in scientific research and clinical practice.

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