Profiling Ubiquitin and Ubiquitin-like Dependent Post-translational Modifications and Identification of Significant Alterations. J Vis Exp. 2019 Nov 07;(153): Authors: Swayden M, Dobric A, Berthois Y, Villalba H, Audebert S, Camoin L, Iovanna J, Soubeyran P Abstract Ubiquitin (ub) and ubiquitin-like (ubl) dependent post-translational modifications of proteins play fundamental biological regulatory roles within the cell by controlling protein stability,...
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Infecting Mice with Malassezia spp. to Study the Fungus-Host Interaction. J Vis Exp. 2019 Nov 06;(153): Authors: Sparber F, LeibundGut-Landmann S Abstract Animal models are crucial for infectious disease research. They provide an important basis for analyzing the full spectrum of interactions that occur between microbes and their host in vivo in a tissue-specific manner. Pathogenic fungi are increasingly recognized as a serious threat for humans and...
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Analysis of Hematopoietic Stem Progenitor Cell Metabolism. J Vis Exp. 2019 Nov 09;(153): Authors: Scapin G, Goulard MC, Dharampuriya PR, Cillis JL, Shah DI Abstract Hematopoietic stem progenitor cells (HSPCs) have distinct metabolic plasticity, which allows them to transition from their quiescent state to a differentiation state to sustain demands of the blood formation. However, it has been difficult to analyze the metabolic status (mitochondrial respiration...
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Characterization of Sickling During Controlled Automated Deoxygenation with Oxygen Gradient Ektacytometry. J Vis Exp. 2019 Nov 05;(153): Authors: Rab MAE, van Oirschot BA, Bos J, Kanne CK, Sheehan VA, van Beers EJ, van Wijk R Abstract In sickle cell disease (SCD), a single point mutation in the gene coding for beta-globin causes the production of abnormal hemoglobin S (HbS). When deoxygenated, HbS can polymerize, forming rigid rods of hemoglobin, resulting...
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Versatile CO2 Transformations into Complex Products: A One-pot Two-step Strategy. J Vis Exp. 2019 Nov 09;(153): Authors: Desmons S, Zhang D, Mejia Fajardo A, Bontemps S Abstract CO2 transformations using a one-pot two-step method are presented herein. The purpose of the method is to give access to a variety of value-added products and notably to generate chiral carbon centers. The crucial first step consists in the selective double hydroboration of CO2...
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Rapid Manufacturing of Thin Soft Pneumatic Actuators and Robots. J Vis Exp. 2019 Nov 08;(153): Authors: Amiri Moghadam AA, Caprio A, Alaie S, Min JK, Dunham S, Mosadegh B Abstract This protocol describes a method for rapid manufacturing of soft pneumatic actuators and robots with an ultrathin form factor using a heat press and a laser cutter machine. The method starts with the lamination of thermoplastic polyurethane (TPU) sheets using a heat press for...
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Fabrication and Design of Wood-Based High-Performance Composites. J Vis Exp. 2019 Nov 09;(153): Authors: Frey M, Zirkelbach M, Dransfeld C, Faude E, Trachsel E, Hannus M, Burgert I, Keplinger T Abstract Delignified densified wood is a new promising and sustainable material that possesses the potential to replace synthetic materials, such as glass fiber reinforced composites, due to its excellent mechanical properties. Delignified wood, however, is rather...
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Cochlear Surface Preparation in the Adult Mouse. J Vis Exp. 2019 Nov 06;(153): Authors: Fang QJ, Wu F, Chai R, Sha SH Abstract Auditory processing in the cochlea depends on the integrity of the mechanosensory hair cells. Over a lifetime, hearing loss can be acquired from numerous etiologies such as exposure to excessive noise, the use of ototoxic medications, bacterial or viral ear infections, head injuries, and the aging process. Loss of sensory hair...
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A Pre-clinical Rat Model for the Study of Ischemia-reperfusion Injury in Reconstructive Microsurgery. J Vis Exp. 2019 Nov 08;(153): Authors: Ballestín A, Casado JG, Abellán E, Vela FJ, Campos JL, Martínez-Chacón G, Bote J, Blázquez R, Sánchez-Margallo FM Abstract Ischemia-reperfusion injury is the main cause of flap failure in reconstructive microsurgery. The rat is the preferred preclinical animal model in many areas of biomedical research due to its...
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Visualization of Candida albicans in the Murine Gastrointestinal Tract Using Fluorescent In Situ Hybridization. J Vis Exp. 2019 Nov 05;(153): Authors: Witchley JN, Penumetcha PM, Noble SM Abstract Candida albicans is a fungal component of the gut microbiota in humans and many other mammals. Although C. albicans does not cause symptoms in most colonized hosts, the commensal reservoir does serve as a repository for infectious disease, and the presence...
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Oligomerization Dynamics of Cell Surface Receptors in Living Cells by Total Internal Reflection Fluorescence Microscopy Combined with Number and Brightness Analysis. J Vis Exp. 2019 Nov 06;(153): Authors: Zamai M, Trullo A, Arza E, Cavallaro U, Caiolfa VR Abstract Despite the importance and ubiquity of receptor oligomerization, few methods are applicable for detecting clustering events and measuring the degree of clustering. Here, we describe an imaging...
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Highly Efficient Transfection of Primary Macrophages with In Vitro Transcribed mRNA. J Vis Exp. 2019 Nov 09;(153): Authors: Herb M, Farid A, Gluschko A, Krönke M, Schramm M Abstract Macrophages are phagocytic cells specialized in detecting molecules of non-self origin. To this end, they are equipped with a large array of pattern recognition receptors (PRRs). Unfortunately, this also makes macrophages particularly challenging to transfect as the transfection...
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Transcranial Direct Current Stimulation for Online Gamers. J Vis Exp. 2019 Nov 09;(153): Authors: Lee SH, Im JJ, Oh JK, Choi EK, Yoon S, Bikson M, Song IU, Jeong H, Chung YA Abstract Transcranial direct current stimulation (tDCS) is a noninvasive brain stimulation technique that applies a weak electric current to the scalp to modulate neuronal membrane potentials. Compared to other brain stimulation methods, tDCS is relatively safe, simple, and inexpensive...
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Inducing Meningococcal Meningitis Serogroup C in Mice via Intracisternal Delivery. J Vis Exp. 2019 Nov 05;(153): Authors: Pagliuca C, Scaglione E, Carraturo F, Mantova G, Marino MM, Pishbin MV, Pagliarulo C, Colicchio R, Salvatore P Abstract Neisseria meningitidis (meningococcus) is a narrow-host-range microorganism, globally recognized as the leading cause of bacterial meningitis. Meningococcus is a transient colonizer of human nasopharynx of approximately...
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Comparative Lesions Analysis Through a Targeted Sequencing Approach. J Vis Exp. 2019 Nov 05;(153): Authors: Vicentini C, Mafficini A, Simbolo M, Fassan M, Delfino P, Lawlor RT, Rusev B, Scarpa A, Corbo V Abstract Assessing intra-tumoral heterogeneity (ITH) is of paramount importance to anticipate failure of targeted therapies and design accordingly effective anti-tumor strategies. Although concerns are frequently raised due to differences in sample processing...
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Short-Term Free-Floating Slice Cultures from the Adult Human Brain. J Vis Exp. 2019 Nov 05;(153): Authors: Fernandes A, Mendes ND, Almeida GM, Nogueira GO, Machado CM, Horta-Junior JAC, Assirati Junior JA, Garcia-Cairasco N, Neder L, Sebollela A Abstract Organotypic, or slice cultures, have been widely employed to model aspects of the central nervous system functioning in vitro. Despite the potential of slice cultures in neuroscience, studies using adult...
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Generation of a Liver Orthotopic Human Uveal Melanoma Xenograft Platform in Immunodeficient Mice. J Vis Exp. 2019 Nov 06;(153): Authors: Kageyama K, Ozaki S, Sato T Abstract In recent decades, subcutaneously implanted patient-derived xenograft tumors or cultured human cell lines have been increasingly recognized as more representative models to study human cancers in immunodeficient mice than traditional established human cell lines in vitro. Recently,...
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A Preclinical Model to Assess Brain Recovery After Acute Stroke in Rats. J Vis Exp. 2019 Nov 06;(153): Authors: Liu P, Song XC, Yang XS, Cao QL, Tang YY, Liu XD, Yang M, An WQ, Dong BX, Song XY Abstract The purpose of this study was to establish and validate an animal brain ischemia model in the recovery and sequela stages. A middle cerebral artery occlusion/reperfusion (MCAO/R) model in male Sprague-Dawley rats was chosen. By changing the rat's weight...
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Optocardiography and Electrophysiology Studies of Ex Vivo Langendorff-perfused Hearts. J Vis Exp. 2019 Nov 07;(153): Authors: Swift LM, Jaimes R, McCullough D, Burke M, Reilly M, Maeda T, Zhang H, Ishibashi N, Rogers JM, Posnack NG Abstract Small animal models are most commonly used in cardiovascular research due to the availability of genetically modified species and lower cost compared to larger animals. Yet, larger mammals are better suited for translational...
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A Human 3D Extracellular Matrix-Adipocyte Culture Model for Studying Matrix-Cell Metabolic Crosstalk. J Vis Exp. 2019 Nov 07;(153): Authors: Flesher CG, Baker NA, Strieder-Barboza C, Polsinelli D, Webster PJ, Varban OA, Lumeng CN, O'Rourke RW Abstract The extracellular matrix (ECM) plays a central role in regulating tissue homeostasis, engaging in crosstalk with cells and regulating multiple aspects of cellular function. The ECM plays a particularly...
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