University of Missouri, Kansas City View Institution's Website 5 articles published in JoVE Biology RNA-seq Analysis of Transcriptomes in Thrombin-treated and Control Human Pulmonary Microvascular Endothelial Cells Dilyara Cheranova1, Margaret Gibson1, Suman Chaudhary1, Li Qin Zhang1, Daniel P. Heruth1, Dmitry N. Grigoryev1, Shui Qing Ye1 1Children's Mercy Hospital and Clinics, School of Medicine, University of Missouri-Kansas City This protocol presents a complete and detailed procedure to apply RNA-seq, a powerful next-generation DNA sequencing technology, to profile transcriptomes in human pulmonary microvascular endothelial cells with or without thrombin treatment. This protocol is generalizable to various cells or tissues affected by different reagents or disease states. Biology Ex Vivo Assessment of Contractility, Fatigability and Alternans in Isolated Skeletal Muscles Ki Ho Park1, Leticia Brotto2, Oanh Lehoang1, Marco Brotto2, Jianjie Ma1, Xiaoli Zhao1,3 1Department of Physiology and Biophysics, UMDNJ-Robert Wood Johnson Medical School, 2Muscle Biology Research Group, University of Missouri-Kansas City, 3Pharmacology division, College of Pharmacy, DHLRI, Ohio State University We describe a method to directly measure muscle force, muscle power, contractile kinetics and fatigability of isolated skeletal muscles in an in vitro system using field stimulation. Valuable information on Ca2+ handling properties and contractile machinery of the muscle can be obtained using different stimulating protocols. Medicine The α-test: Rapid Cell-free CD4 Enumeration Using Whole Saliva Cynthia L. Bristow1, Mariya A. Babayeva1, Rozbeh Modarresi1, Carole P. McArthur2, Santosh Kumar3, Charles Awasom4, Leo Ayuk4, Annette Njinda5, Paul Achu5, Ronald Winston6 1Department of Medicine, Weill Cornell Medical College, 2Department of Oral Biology, University of Missouri-Kansas City-School of Dentistry, 3Department of Pharmacology and Toxicology, University of Missouri Kansas City- School of Pharmacy, 4Regional Hospital, Bamenda, NWP, Cameroon, 5Mezam Polyclinic HIV/AIDS Treatment Center, Cameroon, 6Institute for Human Genetics and Biochemistry A CD4 enumeration method, the α-test, is described which uses whole saliva to provide rapid and accurate CD4 counts. The α-test costs pennies and eliminates the need for technical training, costly reagents such as monoclonal antibodies, instrumentation, refrigeration, transport of samples, as well as collection and handling of blood. Biology Fluorescence-based Measurement of Store-operated Calcium Entry in Live Cells: from Cultured Cancer Cell to Skeletal Muscle Fiber Zui Pan1, Xiaoli Zhao2, Marco Brotto3 1Department of Physiology and Biophysics, Confocal Microscopy and Cell Imaging Core, Robert Wood Johnson Medical School, 2Department of Physiology and Biophysics, Robert Wood Johnson Medical School, 3Muscle Biology Research Group-MUBIG Schools of Nursing & Medicine, University of Missouri-Kansas City The extent of store-operated Ca2+ entry (SOCE) can be monitored using fluorescent Ca2+ indicators. Mn2+ quenching of such indicators assays SOCE in cultured cells and skeletal muscle fibers. A technique allowing spatial and temporal resolution of SOCE by confocal imaging of mechanically skinned muscle fibers is also described. Biology Yeast Colony Embedding Method Sarah Piccirillo1, Saul M. Honigberg1 1School of Biological Sciences, University of Missouri - Kansas City A method for embedding yeast colonies allowing sectioning for light and electron microscopy. This protocol allows determination of the distribution of sporulated cells and pseudohyphal cells within colonies providing a new tool toward understanding the organization of cell types within a fungal community.
