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Johns Hopkins University Bloomberg School of Public Health

11 ARTICLES PUBLISHED IN JoVE

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Medicine

A Simple Method of Mouse Lung Intubation
Sandhya Das 1, Kelvin MacDonald 2, Herng-Yu Sucie Chang 1, Wayne Mitzner 1
1Department of Environmental Health Sciences, Program in Respiratory Biology and Lung Disease, Johns Hopkins Bloomberg School of Public Health, 2Department of Pediatrics, Oregon Health Sciences University

This paper describes a striaghforward and efficient method of intubating mice for pulmonary function measurements or pulmonary instillation, that allows the mice to recover and be studied at later times. The procedure involves an inexpensive fiberoptic light source that directly illuminates the trachea.

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JoVE Core

Collecting Saliva and Measuring Salivary Cortisol and Alpha-amylase in Frail Community Residing Older Adults via Family Caregivers
Nancy A. Hodgson 1, Douglas A. Granger 2,3,4
1Center for Innovative Care in Aging, Johns Hopkins University School of Nursing, 2Institute for Interdisciplinary Salivary Bioscience Research, Arizona State University, 3Johns Hopkins University School of Nursing, 4Johns Hopkins University Bloomberg School of Public Health

We demonstrate: (1) procedures for collection of salivary samples in cognitive impaired older adults by family caregivers in the home setting, (2) procedures for measuring stress activity via salivary cortisol and alpha amylase, and (3) representative profiles. Protocols that allow researchers to study stress-linked processes advance our understanding of biological sensitivity and susceptibility.

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Biology

Strategies for Tracking Anastasis, A Cell Survival Phenomenon that Reverses Apoptosis
Ho Lam Tang 1,2,3, Ho Man Tang 1,2,3, J. Marie Hardwick 1, Ming Chiu Fung 2
1W. Harry Feinstone Department of Molecular Microbiology and Immunology, Johns Hopkins University Bloomberg School of Public Health, 2School of Life Sciences, Chinese University of Hong Kong, 3Center for Cell Dynamics, Department of Biological Chemistry, Johns Hopkins University School of Medicine

The term anastasis refers to the phenomenon in which dying cells reverse a cell suicide process at a late stage, repair themselves, and ultimately survive. Here we demonstrate protocols for detecting and tracking cells that undergo anastasis.

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Biology

Phenotyping Mouse Pulmonary Function In Vivo with the Lung Diffusing Capacity
Nathachit Limjunyawong 1, Jonathan Fallica 1, Amritha Ramakrishnan 2, Kausik Datta 3, Matthew Gabrielson 1, Maureen Horton 1, Wayne Mitzner 1
1Department of Environmental Health Sciences, Johns Hopkins University Bloomberg School of Public Health, 2Department of International Health, Johns Hopkins University Bloomberg School of Public Health, 3Department of Medicine, Johns Hopkins University School of Medicine

We describe a means to quickly and simply measure the lung diffusing capacity in mice and show that it is sufficiently sensitive to phenotype changes in multiple common lung pathologies. This metric thus brings direct translational relevance to the mouse models, since diffusing capacity is also easily measured in humans.

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JoVE Core

Measurement of the Pressure-volume Curve in Mouse Lungs
Nathachit Limjunyawong 1, Jonathan Fallica 1, Maureen R. Horton 1, Wayne Mitzner 1
1Environmental Health Sciences, Bloomberg School of Public Health, Johns Hopkins University

Here we present a protocol to simply and reliably measure the lung pressure-volume curve in mice, showing that it is sufficiently sensitive to detect phenotypic parenchymal changes in two common lung pathologies, pulmonary fibrosis and emphysema. This metric provides a means to quantify the lung’s structural changes with developing pathology.

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Medicine

Instillation and Fixation Methods Useful in Mouse Lung Cancer Research
Nathachit Limjunyawong 1, Jason Mock 2, Wayne Mitzner 1
1Bloomberg School of Public Health, Environmental Health Sciences, Johns Hopkins University, 2Department of Medicine, Pulmonary Diseases and Critical Care Medicine, University of North Carolina School of Medicine

The goal of this paper is to describe simple methods that will greatly aid in the setup and analysis of mouse lungs with lung cancer or other pathologies. We present 3 protocols to simply and reliably carry out lung instillations, fixation, and lung volume measurements.

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Biology

In Vivo Biosensor Tracks Non-apoptotic Caspase Activity in Drosophila
Ho Lam Tang 1, Ho Man Tang 1, Ming Chiu Fung 2, J. Marie Hardwick 1
1W. Harry Feinstone Department of Molecular Microbiology and Immunology, Johns Hopkins University Bloomberg School of Public Health, 2School of Life Sciences, Chinese University of Hong Kong

To detect healthy cells in whole animals that contain low levels of caspase activity, the highly sensitive biosensor designated CaspaseTracker was generated for Drosophila. Caspase-dependent biosensor activity is detected in long-lived healthy cells throughout the internal organs of adult animals reared under optimized conditions in the absence of death stimuli.

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Medicine

Detecting Anastasis In Vivo by CaspaseTracker Biosensor
Ho Man Tang 1,2, Ming Chiu Fung 2, Ho Lam Tang 3
1Institute for Basic Biomedical Sciences, Johns Hopkins University School of Medicine, 2School of Life Sciences, Chinese University of Hong Kong, 3Department of Neurosurgery, Johns Hopkins University School of Medicine

Anastasis is technically challenging to detect in vivo because the cells that have reversed the cell death process can be morphologically indistinguishable from normal healthy cells. Here we describe protocols for detecting and tracking cells that undergo anastasis in live animals by using our newly developed in vivo CaspaseTracker biosensor system.

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Developmental Biology

A Seminiferous Tubule Squash Technique for the Cytological Analysis of Spermatogenesis Using the Mouse Model
Stephen R Wellard *1, Jessica Hopkins *1, Philip W. Jordan 1
1Department of Biochemistry and Molecular Biology, Johns Hopkins Bloomberg School of Public Health

The goal of this tubule squash technique is to rapidly assess cytological features of developing mouse spermatocytes while preserving cellular integrity. This method allows for the study of all stages of spermatogenesis, and can be easily implemented alongside other biochemical and molecular biological approaches for the study of mouse meiosis.

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Education

Chromatin Spread Preparations for the Analysis of Mouse Oocyte Progression from Prophase to Metaphase II
Grace H. Hwang *1, Jessica L. Hopkins *1, Philip W. Jordan 1
1Department of Biochemistry and Molecular Biology, Johns Hopkins University Bloomberg School of Public Health

Oogenesis in mammals is known to be error-prone, particularly due to chromosome missegregation. This manuscript describes chromatin spread preparation methods for mouse prophase, metaphase I and II-staged oocytes. These fundamental techniques allow for the study of chromatin-bound proteins and chromosome morphology throughout mammalian oogenesis.

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Immunology and Infection

High-Speed Human Temporal Bone Sectioning for the Assessment of COVID-19-Associated Middle Ear Pathology
Nicholas S. Andresen 1, Megan K. Wood 2, Daniela Čiháková 3, C. Matthew Stewart 1
1Department of Otolaryngology - Head & Neck Surgery, Johns Hopkins University School of Medicine, 2W. Harry Feinstone Department of Molecular Microbiology and Immunology, Johns Hopkins University Bloomberg School of Public Health, 3Department of Pathology, Johns Hopkins University School of Medicine

This article describes a technique for rapid human temporal bone sectioning that utilizes a microsaw with twin diamond blades to generate thin slices for rapid decalcification and analysis of temporal bone immunohistochemistry.

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