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Pennsylvania State University

16 ARTICLES PUBLISHED IN JoVE

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Neuroscience

Low-stress Route Learning Using the Lashley III Maze in Mice
Amanda Bressler 1, David Blizard 2, Anne Andrews 3,4,5,6
1Department of Chemistry, Pennsylvania State University, 2Center for Developmental and Health Genetics, Pennsylvania State University, 3Department of Veterinary and Biomedical Sciences, Pennsylvania State University, 4Huck Institute of the Life Sciences, Pennsylvania State University, 5California NanoSystems Institute, University of California, Los Angeles, 6Semel Institute of Neuroscience and Human Behavior, University of California, Los Angeles

The Lashley III maze is a route-learning task that does not rely on aversive stimuli or visual cues. It is thus a highly attractive option for evaluating learning and memory, especially in aging mice or otherwise where stress is a consideration.

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Biology

Primer-Free Aptamer Selection Using A Random DNA Library
Weihua Pan 1, Ping Xin 1, Susan Patrick 1, Stacey Dean 2, Christine Keating 2, Gary Clawson 3,4
1Department of Pathology, Hershey Medical Center, Pennsylvania State University, 2Department of Chemistry, Pennsylvania State University, 3Departments of Pathology, and Biochemistry and Molecular Biology, Hershey Medical Center, Pennsylvania State University, 4Materials Research Institute, Pennsylvania State University

SELEX protocols comprise multiple rounds of selection, each of which require regeneration of bound ligands, which in turn require fixed primer sequences flanking the random library regions. These fixed primer sequences can interfere with the selection process (false positives and negatives). Here we present a primer-free protocol.

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Basic Biology

Electrical Safety
Robert Rioux 1, Suprita Jharimune 1
1Pennsylvania State University

Electrical Safety

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Medicine

Dried Blood Spot Collection of Health Biomarkers to Maximize Participation in Population Studies
Michael W. Ostler 1, James H. Porter 1,2, Orfeu M. Buxton 1,2,3,4
1Center for Population and Development Studies, Harvard School of Public Health, 2Department of Medicine, Brigham and Women's Hospital, 3Division of Sleep Medicine, Harvard Medical School, 4Department of Biobehavioral Health, Pennsylvania State University

Biomarkers are directly-measured biological indicators of disease or health. In population and social sciences, biomarkers need to be easy to obtain, transport, and analyze. Dried Blood Spot (DBS) collection meets this need, can be collected in the field with high response rates and analyzed for a variety of biomarkers.

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Bioengineering

Molecular Entanglement and Electrospinnability of Biopolymers
Lingyan Kong 1, Gregory R. Ziegler 1
1Department of Food Science, Pennsylvania State University

Electrospinning is a fascinating technique used to fabricate micro- to nano-scale fibers from a wide variety of materials. Molecular entanglement of the constituent polymers in the spinning dope is essential for successful electrospinning. We present a protocol for utilizing rheology to evaluate the electrospinnability of two biopolymers, starch and pullulan.

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Environment

The Use of an Automated System (GreenFeed) to Monitor Enteric Methane and Carbon Dioxide Emissions from Ruminant Animals
Alexander N. Hristov 1, Joonpyo Oh 1, Fabio Giallongo 1, Tyler Frederick 1, Holley Weeks 1, Patrick R. Zimmerman 2, Michael T. Harper 1, Rada A. Hristova 3, R. Scott Zimmerman 2, Antonio F. Branco 4
1Department of Animal Science, Pennsylvania State University, 2C-Lock, Inc. Rapid City, SD, 3Color Productions, State College, PA, 4Departamento de Zootecnia, Universidade Estadual de Maringá

Accuracy and precision of the techniques used to measure methane emissions from ruminant animals are critically important for the success of greenhouse gas mitigation efforts. This manuscript describes the principles and operation of an automated system to monitor methane and carbon dioxide mass fluxes from the breath of ruminant animals.

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Immunology and Infection

Inoculating Anopheles gambiae Mosquitoes with Beads to Induce and Measure the Melanization Immune Response
Antoine M. G. Barreaux 1,2, Priscille Barreaux 1,2, Matthew B. Thomas 2, Jacob C. Koella 1
1Laboratoire d'écologie et d'épidémioloigie parasitaire, Institut de biologie, Université de Neuchâtel, 2Merkle Lab, Department of Entomology, Pennsylvania State University

Through inoculation with beads, the described technique enables the stimulation of the mosquito melanization response in the hemolymph circulating system. The amount of melanin covering the beads can be measured after dissection as a measure of the immune response.

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Immunology and Infection

Isolation, Characterization, and Purification of Macrophages from Tissues Affected by Obesity-related Inflammation
Joselyn N. Allen 1, Adwitia Dey 1, Ruth Nissly 2, James Fraser 1, Shan Yu 1, Gayathri Balandaram 1, Jeffrey M. Peters 1, Pamela A. Hankey-Giblin 1
1Department of Veterinary and Biomedical Sciences, The Pennsylvania State University, 2Microscopy and Cytometry Facility, The Huck Institutes of Life Sciences, Pennsylvania State University

This protocol allows a researcher to isolate and characterize tissue-resident macrophages in various hallmark inflamed tissues extracted from diet-induced models of metabolic disorders.

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Environment

Continuous Hydrologic and Water Quality Monitoring of Vernal Ponds
Odette Mina 1,2, Heather E. Gall 1, Joseph W. Chandler 1, Jeremy Harper 3, Malcolm Taylor 1
1Department of Agricultural and Biological Engineering, Pennsylvania State University, 2Energy and Environmental Sustainability Laboratory, Pennsylvania State University, 3Department of Ecosystem Science and Management, Pennsylvania State University

Understanding the ecosystem services and processes provided by vernal ponds and the impacts of anthropogenic activities on their ability to provide these services requires intensive hydrologic monitoring. This sampling protocol using in-situ monitoring equipment was developed to understand the impact of anthropogenic activities on water levels and quality.

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Immunology and Infection

Enrichment of Bacterial Lipoproteins and Preparation of N-terminal Lipopeptides for Structural Determination by Mass Spectrometry
Krista M. Armbruster 1, Timothy C. Meredith 1
1Department of Biochemistry and Molecular Biology, Pennsylvania State University

The enrichment of bacterial lipoproteins using a non-ionic surfactant phase partitioning method is described for direct use in TLR assays or other applications. Further steps are detailed to prepare N-terminal tryptic lipopeptides for structural characterization by mass spectrometry.

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Environment

Wastewater Irrigation Impacts on Soil Hydraulic Conductivity: Coupled Field Sampling and Laboratory Determination of Saturated Hydraulic Conductivity
Jack E. Watson 1, Tyson Robb 2, Danielle Andrews-Brown 3, Melissa Miller 1
1Department of Ecosystem Science and Management, Pennsylvania State University, 2Department of Geography and Environmental Sustainability, State University of New York, Oneonta, 3Department of Geology and Environmental Science, University of Pittsburgh

Here we present a methodology which matches a soil sample size and a hydraulic conductivity measurement device to prevent the so-called wall flow along the inside of the soil container from being erroneously included in water flow measurements. Its use is demonstrated with samples collected from a wastewater irrigation site.

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Biochemistry

Kinase Inhibitor Screening In Self-assembled Human Protein Microarrays
Fernanda Festa 1,2, Joshua Labaer 1
1Virginia G. Piper Center for Personalized Diagnostics, Biodesign Institute, Arizona State University, 2Department of Pediatrics, and Biochemistry/Molecular Biology, College of Medicine, Pennsylvania State University

A detailed protocol for the generation of self-assembled human protein microarrays for the screening of kinase inhibitors is presented.

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Behavior

A Standardized Protocol for Preference Testing to Assess Fish Welfare
Cairsty DePasquale 1, Jennifer Sturgill 1, Victoria A. Braithwaite 2
1Department of Biology, Pennsylvania State University - Altoona, 2Department of Ecosystem Science & Management, Pennsylvania State University

A fundamental aspect of assessing the welfare of animals in captivity is to ask whether the animals have what they want. Here, we present a protocol to determine housing preference in the zebrafish (Danio rerio) with respect to the presence/absence of environmental enrichment and access to flowing of water.

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Biochemistry

Advancing High-Resolution Imaging of Virus Assemblies in Liquid and Ice
Liza-Anastasia DiCecco *1,2, Samantha Berry *1, G. M. Jonaid *1,3, Maria J. Solares 1,4, Liam Kaylor 1,4, Jennifer L. Gray 5, Carol Bator 6, William J. Dearnaley 1, Michael Spilman 7, Madeline J. Dressel-Dukes 8, Kathryn Grandfield 2, Sarah M. McDonald Esstman 9, Deborah F. Kelly 1,5,6
1Department of Biomedical Engineering, Pennsylvania State University, 2Department of Materials Science and Engineering, McMaster University, 3Bioinformatics and Genomics Graduate Program, Huck Institutes of the Life Sciences, Pennsylvania State University, 4Molecular, Cellular, and Integrative Biosciences Graduate Program, Huck Institutes of the Life Sciences, Pennsylvania State University, 5Materials Research Institute, Pennsylvania State University, 6Huck Institutes of the Life Sciences, Pennsylvania State University, 7Applications team, Direct Electron, 8Application Scientist, Protochips, Inc., 9Department of Biology, Wake Forest University

Here protocols are described to prepare virus assemblies suitable for liquid-EM and cryo-EM analysis at the nanoscale using transmission electron microscopy.

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Engineering

Measuring Local Tissue Strains in Tendons via Open-Source Digital Image Correlation
Stanton Godshall 1, Krishna Pedaprolu 1, Erica Vasti 1, Faezeh Eskandari 2, Spencer E. Szczesny 1,3
1Department of Biomedical Engineering, Pennsylvania State University, 2Department of Biomedical Engineering, Amirkabir University of Technology, 3Department of Orthopaedics and Rehabilitation, Pennsylvania State University

This paper describes an open-source digital image correlation algorithm for measuring local 2D tissue strains within tendon explants. The accuracy of the technique has been validated using multiple techniques, and it is available for public use.

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Musculoskeletal Research: Current Methods and Innovative Approaches
Reyad A. Elbarbary 1,2,3,4
1Center for Orthopaedic Research and Translational Science (CORTS), The Pennsylvania State University College of Medicine, 2Department of Orthopaedics and Rehabilitation, The Pennsylvania State University College of Medicine, 3Department of Biochemistry and Molecular Biology, The Pennsylvania State University College of Medicine, 4Center for RNA Molecular Biology, Pennsylvania State University

Musculoskeletal Research: Current Methods and Innovative Approaches

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