Protocol
Production of Synthetic Nuclear Melt Glass
Joshua J. Molgaard1, John D. Auxier II2,3, Andrew V. Giminaro2,3, Colton J. Oldham2, Jonathan Gill2, Howard L. Hall2,3,4
1Department of Physics and Nuclear Engineering, United States Military Academy, 2Department of Nuclear Engineering, University of Tennessee, 3Radiochemistry Center of Excellence (RCoE), University of Tennessee, 4Institute for Nuclear Security, University of Tennessee
A protocol for the production of synthetic nuclear melt glass, similar to trinitite, is presented.
A Simple Method for the Size Controlled Synthesis of Stable Oligomeric Clusters of Gold Nanoparticles under Ambient Conditions
Marlon Lawrence1, Anze Testen1, Tilen Koklic2, Oliver Smithies1
1Department of Pathology and Laboratory Medicine, University of North Carolina at Chapel Hill, 2Condensed Matter Physics Department, Laboratory of Biophysics, Jozef Stefan Institute
We describe a simple method for producing highly stable oligomeric clusters of gold nanoparticles via the reduction of chloroauric acid (HAuCl4) with sodium thiocyanate (NaSCN). The oligoclusters have a narrow size distribution and can be produced with a wide range of sizes and surface coats.
RNAi Trigger Delivery into Anopheles gambiae Pupae
Kimberly Regna1, Rachel M. Harrison1, Shannon A. Heyse1, Thomas C. Chiles1, Kristin Michel2, Marc A. T. Muskavitch1,3
1Biology Department, Boston College, 2Division of Biology, Kansas State University, 3Discovery Research, Biogen
RNA interference (RNAi) is an extremely valuable tool for uncovering gene function. However, the ability to target genes using RNAi during pre-adult stages is limited in the major human malaria vector Anopheles gambiae. We describe an RNAi protocol to reduce gene function via direct injection during pupal development.
Spotting Cheetahs: Identifying Individuals by Their Footprints
Zoe C. Jewell1, Sky K. Alibhai1, Florian Weise2,3, Stuart Munro2, Marlice Van Vuuren4, Rudie Van Vuuren4
1WildTrack and Nicholas School of the Environment, Duke University, 2N∕a′an ku sê Research Programme, 3Division of Biology and Conservation Ecology, School of Science and the Environment, Manchester Metropolitan University, 4N∕a′an ku sê Foundation
The cheetah (Acinonyx jubatus) is an iconic, endangered species, but conservation efforts are challenged by habitat shrinkage and conflict with commercial farmers. The footprint identification technique, a robust, accurate and cost-effective image classification system, is a new approach to monitoring cheetahs.
A Novel Behavioral Assay to Investigate Gustatory Responses of Individual, Freely-moving Bumble Bees (Bombus terrestris)
Carolyn Ma, Sébastien Kessler, Alexander Simpson, Geraldine Wright
Institute of Neuroscience, Newcastle University
A novel behavioral assay is described for investigating the short term gustatory responses of the mouthparts of freely-moving bumble bees (Bombus terrestris) toward nutrients and toxins in solution.
Intracoronary Acetylcholine Provocation Testing for Assessment of Coronary Vasomotor Disorders
Peter Ong, Anastasios Athanasiadis, Udo Sechtem
Department of Cardiology, Robert-Bosch-Krankenhaus
Intracoronary acetylcholine testing has been established for the assessment of epicardial coronary spasm more than 30 years ago. Recently, the focus has shifted towards the microcirculation and it has been shown that microvascular spasm can be detected using ACH-testing. This article describes the ACH-test and its implementation in daily routine.
A Mimic of the Tumor Microenvironment: A Simple Method for Generating Enriched Cell Populations and Investigating Intercellular Communication
Jason D. Domogauer, Sonia M. de Toledo, Edouard I. Azzam
Department of Radiology, New Jersey Medical School, Rutgers University
We adapted a permeable microporous membrane insert to mimic the tumor microenvironment (TME). The model consists of a mixed cell culture, allows simplified generation of highly enriched individual cell populations without using fluorescent tagging or cell sorting, and permits studying intercellular communication within the TME under normal or stress conditions.
Genetic Engineering of an Unconventional Yeast for Renewable Biofuel and Biochemical Production
Ai-Qun Yu1,2, Nina Pratomo1,2, Tee-Kheang Ng1,2, Hua Ling1,2, Han-Saem Cho1,2, Susanna Su Jan Leong1,2,3, Matthew Wook Chang1,2
1Department of Biochemistry, Yong Loo Lin School of Medicine, National University of Singapore, 2NUS Synthetic Biology for Clinical and Technological Innovation (SynCTI), Life Sciences Institute, National University of Singapore, 3Food Science and Chemical Engineering, Singapore Institute of Technology
We herein report methods on the molecular genetic manipulation of the Yarrowia lipolytica Po1g strain for improved gene deletion efficiency. The resulting engineered Y. lipolytica strains have potential applications in biofuel and biochemical production.
Species Determination and Quantitation in Mixtures Using MRM Mass Spectrometry of Peptides Applied to Meat Authentication
Yvonne Gunning1, Andrew D. Watson1, Neil M. Rigby2, Mark Philo1, Joshua K. Peazer1,3, E. Kate Kemsley1
1Analytical Sciences Unit, Institute of Food Research, 2Institute of Food Research, 3School of Chemistry, University of East Anglia
We present a protocol for identifying and quantifying the components in mixtures of species possessing similar proteins. Mass spectrometry detects peptides for identification, and gives relative quantitation by ratios of peak areas. As a tool food for fraud detection, the method can detect 1% horse in beef.
Safety Precautions and Operating Procedures in an (A)BSL-4 Laboratory: 3. Aerobiology
J. Kyle Bohannon, Krisztina Janosko, Michael R. Holbrook, Jason Barr, Daniela Pusl, Laura Bollinger, Linda Coe, Lisa E. Hensley, Peter B. Jahrling, Jiro Wada, Jens H. Kuhn, Matthew G. Lackemeyer
Integrated Research Facility at Frederick, National Institute of Allergy and Infectious Diseases (NIAID), National Institutes of Health (NIH)
As high-consequence pathogens can potentially infect subjects through airborne particles, aerobiology has been increasingly applied in pathogenesis research and medical countermeasure development. We present a detailed visual demonstration of aerobiology procedures during an aerosol challenge in nonhuman primates in an animal biosafety level 4 maximum containment environment.
Safety Precautions and Operating Procedures in an (A)BSL-4 Laboratory: 1. Biosafety Level 4 Suit Laboratory Suite Entry and Exit Procedures
Krisztina Janosko1, Michael R. Holbrook1, Ricky Adams1, Jason Barr1, Laura Bollinger1, Je T'aime Newton2, Corrie Ntiforo2, Linda Coe1, Jiro Wada1, Daniela Pusl1, Peter B. Jahrling1, Jens H. Kuhn1, Matthew G. Lackemeyer1
1Integrated Research Facility at Frederick, National Institute of Allergy and Infectious Diseases (NIAID), National Institutes of Health (NIH), 2Environmental Health and Safety, Biological and Chemical Safety Program, University of Texas Medical Branch
Although researchers are generally knowledgeable about procedures and safety precautions required for biosafety level 1 or 2 (BSL-1/2) experiments, they may not be familiar with experimental procedures in BSL-4 suit laboratories. This article provides a detailed visual demonstration of BSL-4 suit laboratory systems check, laboratory entry, movement, and exit procedures.
Safety Precautions and Operating Procedures in an (A)BSL-4 Laboratory: 2. General Practices
Steven Mazur, Michael R. Holbrook, Tracey Burdette, Nicole Josleyn, Jason Barr, Daniela Pusl, Laura Bollinger, Linda Coe, Peter B. Jahrling, Matthew G. Lackemeyer, Jiro Wada, Jens H. Kuhn, Krisztina Janosko
Integrated Research Facility at Frederick, National Institute of Allergy and Infectious Diseases (NIAID), National Institutes of Health (NIH)
Performing viral assays in a BSL-4 laboratory is more involved compared to work in a BSL-2 laboratory due to required additional safety precautions. Here, we present an overview of practices and procedures used inside a BSL-4 laboratory illustrating proper Class II biosafety cabinet usage, waste management/disposal, and sample removal.
A Buoyancy-based Method of Determining Fat Levels in Drosophila
Kelsey E. Hazegh, Tânia Reis
Department of Medicine, Division of Endocrinology, Metabolism, and Diabetes, University of Colorado Anschutz Medical Campus
Here we present a method to measure organismal fat levels in the third instar (L3) larval stage of Drosophila melanogaster. This method exploits the comparatively low density of fat tissue to differentiate between larvae with altered fat stores. Buoyancy-based analysis is a valuable tool for rapid, reproducible, and economical screening.
Using Pharmacological Manipulation and High-precision Radio Telemetry to Study the Spatial Cognition in Free-ranging Animals
Timothy C. Roth*1, Aaron R. Krochmal*2, William B. Gerwig, IV1, Sage Rush3, Nathaniel T. Simmons2, Jeffery D. Sullivan4, Katrina Wachter5
1Department of Psychology, Franklin and Marshall College, 2Department of Biology, Washington College, 3University of Pennsylvania, 4School of Forestry and Wildlife Sciences, Auburn University, 5Morsani College of Medicine, University of South Florida
This paper describes a novel protocol that combines the pharmacological manipulation of memory and radio telemetry to document and quantify the role of cognition in navigation.
Disclosures
No conflicts of interest declared.
