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Journal of Visualized Experiments: JoVE

Amanda L Forster, Viviana Rodriguez Cardenas, Ajay Krishnamurthy, Zois Tsinas, Amy Engelbrecht-Wiggans, Nolan Gonzalez
Traditionally, soft body armor has been made from poly(p-phenylene terephthalamide) (PPTA) and ultra-high molecular weight polyethylene. However, to diversify the fiber choices in the United States body armor market, copolymer fibers based on the combination of 5-amino-2-(p-aminophenyl) benzimidazole (PBIA) and the more conventional PPTA were introduced. Little is known regarding the long-term stability of these fibers, but as condensation polymers, they are expected to have potential sensitivity to moisture and humidity...
September 1, 2018: Journal of Visualized Experiments: JoVE
Lucy K Goodman, Nicola S Anstice, Suzanne Stevens, Benjamin Thompson, Trecia A Wouldes
Classical eyeblink conditioning (EBC) refers to the learned association between a conditioned stimulus (an auditory tone) and an unconditioned stimulus (a puff of air to the cornea). Eyeblink conditioning is often used experimentally to detect abnormalities in cerebellar-dependent learning and memory that underlies this type of associative learning. While experiments in adults and older children are relatively simple to administer using commercial equipment, eyeblink conditioning in infants is more challenging due to their poor compliance, which makes correct positioning of the equipment difficult...
September 1, 2018: Journal of Visualized Experiments: JoVE
Qiao Li, Cheng Qian, Feng-Quan Zhou
Electroporation is an essential non-viral gene transfection approach to introduce DNA plasmids or small RNA molecules into cells. A sensory neuron in the dorsal root ganglion (DRGs) extends a single axon with two branches. One branch goes to the peripheral nerve (peripheral branch), and the other branch enters the spinal cord through the dorsal root (central branch). After the neural injury, the peripheral branch regenerates robustly whereas the central branch does not regenerate. Due to the high regenerative capacity, sensory axon regeneration has been widely used as a model system to study mammalian axon regeneration in both the peripheral nervous system (PNS) and the central nervous system (CNS)...
September 1, 2018: Journal of Visualized Experiments: JoVE
Ulrich Jarry, Noémie Joalland, Cynthia Chauvin, Béatrice Clemenceau, Claire Pecqueur, Emmanuel Scotet
Glioblastoma multiforme (GBM), the most frequent and aggressive primary brain cancer in adults, is generally associated with a poor prognosis, and scarce efficient therapies have been proposed over the last decade. Among the promising candidates for designing novel therapeutic strategies, cellular immunotherapies have been targeted to eliminate highly invasive and chemo-radioresistant tumor cells, likely involved in a rapid and fatal relapse of this cancer. Thus, administration(s) of allogeneic GBM-reactive immune cell effectors, such as human Vϒ9Vδ2 T lymphocytes, in the vicinity of the tumor would represents a unique opportunity to deliver efficient and highly concentrated therapeutic agents directly into the site of brain malignancies...
September 1, 2018: Journal of Visualized Experiments: JoVE
Stephanie A Michalski, Sangappa B Chadchan, Emily S Jungheim, Ramakrishna Kommagani
The differentiation of human endometrial stromal cells (HESC) from fibroblast-like appearance into secretory decidua is a transformation required for embryo implantation into the uterine lining of the maternal womb. Improper decidualization has been established as a root cause for implantation failure and subsequent early embryo miscarriage. Therefore, understanding the molecular mechanisms underlying decidualization is advantageous to improving the rate of successful births. In vivo based studies of artificial decidualization are often limiting due to ethical dilemmas associated with human research, as well as translational complications within animal models...
September 1, 2018: Journal of Visualized Experiments: JoVE
Athanasios Saragliadis, Thomas Trunk, Jack C Leo
A first approach to study the function of an unknown gene in bacteria is to create a knock-out of this gene. Here, we describe a robust and fast protocol for transferring gene deletion mutations from one Escherichia coli strain to another by using generalized transduction with the bacteriophage P1. This method requires that the mutation be selectable (e.g., based on gene disruptions using antibiotic cassette insertions). Such antibiotic cassettes can be mobilized from a donor strain and introduced into a recipient strain of interest to quickly and easily generate a gene deletion mutant...
September 1, 2018: Journal of Visualized Experiments: JoVE
Keith R Dona, Monika Goss-Varley, Andrew J Shoffstall, Jeffrey R Capadona
We have recently demonstrated that implanting intracortical microelectrodes in the motor corteces of rats results in immediate and lasting motor deficits. Motor impairments were manually quantified through an open field grid test to measure the gross motor function and through a ladder test to measure the fine motor function. Here, we discuss a technique for the automated quantification of the video-recorded tests using our custom Capadona Behavioral Video Analysis System: Grid and Ladder Test, or BVAS. Leveraging simple and readily available coding software (see the Table of Materials), this program allows for the tracking of a single animal on both the open field grid and the ladder tests...
August 31, 2018: Journal of Visualized Experiments: JoVE
Ann M Anderson, Bradford A Bruno, Elizabeth A Donlon, Luisa F Posada, Mary K Carroll
Protocols for preparing and testing catalytic aerogels by incorporating metal species into silica and alumina aerogel platforms are presented. Three preparation methods are described: (a) the incorporation of metal salts into silica or alumina wet gels using an impregnation method; (b) the incorporation of metal salts into alumina wet gels using a co-precursor method; and (c) the addition of metal nanoparticles directly into a silica aerogel precursor mixture. The methods utilize a hydraulic hot press, which allows for rapid (<6 h) supercritical extraction and results in aerogels of low density (0...
August 31, 2018: Journal of Visualized Experiments: JoVE
Nikki Traylor-Knowles
Corals are important ocean invertebrates that are critical for overall ocean health as well as human health. However, due to human impacts such as rising ocean temperatures and ocean acidification, corals are increasingly under threat. To tackle these challenges, advances in cell and molecular biology have proven to be crucial for diagnosing the health of corals. Modifying some of the techniques commonly used in human medicine could greatly improve researchers' ability to treat and save corals. To address this, a protocol for in situ hybridization used primarily in human medicine and evolutionary developmental biology has been adapted for use in adult corals under stress...
August 31, 2018: Journal of Visualized Experiments: JoVE
Jacob M Dixon, Shunji Egusa
The purpose of the presented protocols is to study the process of Au(III) binding to BSA, yielding conformation change-induced red fluorescence (λem = 640 nm) of BSA-Au(III) complexes. The method adjusts the pH to show that the emergence of the red fluorescence is correlated with the pH-induced equilibrium transitions of the BSA conformations. Red fluorescent BSA-Au(III) complexes can only be formed with an adjustment of pH at or above 9.7, which corresponds to the "A-form" conformation of BSA. The protocol to adjust the BSA to Au molar ratio and to monitor the time-course of the process of Au(III) binding is described...
August 31, 2018: Journal of Visualized Experiments: JoVE
Stephanie S L Birnbaum, David C Rinker, Patrick Abbot
Aphids are excellent experimental models for a variety of biological questions ranging from the evolution of symbioses and the development of polyphenisms to questions surrounding insect's interactions with their host plants. Genomic resources are available for several aphid species, and with advances in the next-generation sequencing, transcriptomic studies are being extended to non-model organisms that lack genomes. Furthermore, aphid cultures can be collected from the field and reared in the laboratory for the use in organismal and molecular experiments to bridge the gap between ecological and genetic studies...
August 31, 2018: Journal of Visualized Experiments: JoVE
Ming-Cheng Chen, John R Lake, Keith C Heyde, Warren C Ruder
Microfluidics has become a critical tool in research across the biological, chemical, and physical sciences. One important component of microfluidic experimentation is a stable fluid handling system capable of accurately providing an inlet flow rate or inlet pressure. Here, we have developed a syringe pump system capable of controlling and regulating the inlet fluid pressure delivered to a microfluidic device. This system was designed using low-cost materials and additive manufacturing principles, leveraging three-dimensional (3D) printing of thermoplastic materials and off-the-shelf components whenever possible...
August 30, 2018: Journal of Visualized Experiments: JoVE
Carina Schuster, Sebastian Sterz, Daniel Teupser, Mathias Brügel, Michael Vogeser, Michael Paal
There is an ever-increasing demand for the therapeutic drug monitoring of antibiotics in many clinical facilities, particularly with regard to the implementation of hospital antibiotic stewardship programs. In the current work, we present a multiplex high-performance liquid chromatography-tandem mass spectrometry (HPCL-MS/MS) protocol for the quantification of cefepime, meropenem, ciprofloxacin, moxifloxacin, linezolid, and piperacillin, commonly used antibiotics in intensive care units. The method was previously comprehensively validated according to the guideline of the European Medicines Agency...
August 30, 2018: Journal of Visualized Experiments: JoVE
Eric Gehring, Bruno Bellosi, Ambra Quacchia, Marco Conedera
Dryocosmus kuriphilus Yasumatsu has become a major pest for Castaneasativa since its arrival in Europe. Its galling activity results in the formation of different gall types and prevents the development of normal shoots. Repeated and uncontrolled attacks cause, besides the production of galls and the attendant gall-related reduction in leaf area, progressive corruption of the branch architecture, including the death of branch parts, and an increase in dormant bud activation. Thus far, there have been few attempts to quantify branch architecture damage...
August 30, 2018: Journal of Visualized Experiments: JoVE
Luis Eduardo Trujillo Villanueva, Felipe Legorreta García, Iker Rodrigo Chávez-Urbiola, Francisco Javier Willars-Rodriguez, Rafael Ramírez-Bon, Marius Ramírez-Cardona, Leticia Esperanza Hernández-Cruz, Edgar Arturo Chávez-Urbiola
In this work, the effects on the preparation of bis(thiourea) cadmium chloride crystals when illuminated with ultraviolet (UV) light at a wavelength of 367 nm using the chemical bath deposition technique are studied comparatively. Two experiments are performed to make a comparison: one without UV light and the other with the aid of UV light. Both experiments are performed under equal conditions, at a temperature of 343 K and with a pH of 3.2. The precursors used are cadmium chloride (CdCl2) and thiourea [CS(NH2)2], which are dissolved in 50 mL of deionized water with an acidic pH...
August 30, 2018: Journal of Visualized Experiments: JoVE
Raphael P Weibel, Jascha Grübel, Hantao Zhao, Tyler Thrash, Dario Meloni, Christoph Hölscher, Victor R Schinazi
Virtual reality (VR) experiments are increasingly employed because of their internal and external validity compared to real-world observation and laboratory experiments, respectively. VR is especially useful for geographic visualizations and investigations of spatial behavior. In spatial behavior research, VR provides a platform for studying the relationship between navigation and physiological measures (e.g., skin conductance, heart rate, blood pressure). Specifically, physiological measures allow researchers to address novel questions and constrain previous theories of spatial abilities, strategies, and performance...
August 29, 2018: Journal of Visualized Experiments: JoVE
Ratnesh K Singh, Pablo E Diaz, François Binette, Igor O Nasonkin
The epigenetics of retinal development is a well-studied research field, which promises to bring a new level of understanding about the mechanisms of a variety of human retinal degenerative diseases and pinpoint new treatment approaches. The nuclear architecture of mouse retina is organized in two different patterns: conventional and inverted. Conventional pattern is universal where heterochromatin is localized to the periphery of the nucleus, while active euchromatin resides in the nuclear interior. In contrast, inverted nuclear pattern is unique to the adult rod photoreceptor cell nuclei where heterochromatin localizes to the nuclear center, and euchromatin resides in the nuclear periphery...
August 29, 2018: Journal of Visualized Experiments: JoVE
Ian Restall, Danielle Anne Bozek, Charles Chesnelong, Samuel Weiss, H Artee Luchman
Glioblastoma (GBM) is an aggressive brain tumor that is poorly controlled with the currently available treatment options. Key features of GBMs include rapid proliferation and pervasive invasion into the normal brain. Recurrence is thought to result from the presence of radio- and chemo-resistant brain tumor stem cells (BTSCs) that invade away from the initial cancerous mass and, thus, evade surgical resection. Hence, therapies that target BTSCs and their invasive abilities may improve the otherwise poor prognosis of this disease...
August 29, 2018: Journal of Visualized Experiments: JoVE
Katherine S Moore, Elizabeth A Wiemers, Ariel Kershner, Korissa Belville, Jaimie Jasina, Aziza Ransome, Jessica Avanzato
This method uses a rapid serial visual presentation (RSVP) paradigm to measure the cost of distraction when participants maintain multiple search goals. The protocol identifies two types of distraction within a single task - contingent attentional capture and set-specific capture - that represent different types of limitations of cognitive processing. Participants search for letters in two or more "target" ink colors (e.g., green and orange) within a continuous RSVP stream of heterogeneously colored letters, while ignoring two peripheral RSVPs of letters...
August 29, 2018: Journal of Visualized Experiments: JoVE
Miroslava Stastna, Amandine Thomas, Juliana Germano, Somayeh Pourpirali, Jennifer E Van Eyk, Roberta A Gottlieb
Studies in dynamic changes in protein translation require specialized methods. Here we examined changes in newly-synthesized proteins in response to ischemia and reperfusion using the isolated perfused mouse heart coupled with polysome profiling. To further understand the dynamic changes in protein translation, we characterized the mRNAs that were loaded with cytosolic ribosomes (polyribosomes or polysomes) and also recovered mitochondrial polysomes and compared mRNA and protein distribution in the high-efficiency fractions (numerous ribosomes attached to mRNA), low-efficiency (fewer ribosomes attached) which also included mitochondrial polysomes, and the non-translating fractions...
August 29, 2018: Journal of Visualized Experiments: JoVE
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