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Journal of Visualized Experiments: JoVE

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https://www.readbyqxmd.com/read/28518086/ptr-tof-ms-coupled-with-an-automated-sampling-system-and-tailored-data-analysis-for-food-studies-bioprocess-monitoring-screening-and-nose-space-analysis
#1
Vittorio Capozzi, Sine Yener, Iuliia Khomenko, Brian Farneti, Luca Cappellin, Flavia Gasperi, Matteo Scampicchio, Franco Biasioli
Proton Transfer Reaction (PTR), combined with a Time-of-Flight (ToF) Mass Spectrometer (MS) is an analytical approach based on chemical ionization that belongs to the Direct-Injection Mass Spectrometric (DIMS) technologies. These techniques allow the rapid determination of volatile organic compounds (VOCs), assuring high sensitivity and accuracy. In general, PTR-MS requires neither sample preparation nor sample destruction, allowing real time and non-invasive analysis of samples. PTR-MS are exploited in many fields, from environmental and atmospheric chemistry to medical and biological sciences...
May 11, 2017: Journal of Visualized Experiments: JoVE
https://www.readbyqxmd.com/read/28518085/analysis-of-chromosome-segregation-histone-acetylation-and-spindle-morphology-in-horse-oocytes
#2
Federica Franciosi, Irene Tessaro, Rozenn Dalbies-Tran, Cecile Douet, Fabrice Reigner, Stefan Deleuze, Pascal Papillier, Ileana Miclea, Valentina Lodde, Alberto M Luciano, Ghylene Goudet
The field of assisted reproduction has been developed to treat infertility in women, companion animals, and endangered species. In the horse, assisted reproduction also allows for the production of embryos from high performers without interrupting their sports career and contributes to an increase in the number of foals from mares of high genetic value. The present manuscript describes the procedures used for collecting immature and mature oocytes from horse ovaries using ovum pick-up (OPU). These oocytes were then used to investigate the incidence of aneuploidy by adapting a protocol previously developed in mice...
May 11, 2017: Journal of Visualized Experiments: JoVE
https://www.readbyqxmd.com/read/28518077/inkjet-printed-polyvinyl-alcohol-multilayers
#3
Iulia Salaoru, Zuoxin Zhou, Peter Morris, Gregory J Gibbons
Inkjet printing is a modern method for polymer processing, and in this work, we demonstrate that this technology is capable of producing polyvinyl alcohol (PVOH) multilayer structures. A polyvinyl alcohol aqueous solution was formulated. The intrinsic properties of the ink, such as surface tension, viscosity, pH, and time stability, were investigated. The PVOH-based ink was a neutral solution (pH 6.7) with a surface tension of 39.3 mN/m and a viscosity of 7.5 cP. The ink displayed pseudoplastic (non-Newtonian shear thinning) behavior at low shear rates, and overall, it demonstrated good time stability...
May 11, 2017: Journal of Visualized Experiments: JoVE
https://www.readbyqxmd.com/read/28518076/facile-preparation-of-ultrafine-aluminum-hydroxide-particles-with-or-without-mesoporous-mcm-41-in-ambient-environments
#4
Viktor Dubovoy, Ravi Subramanyam, Michael Stranick, Laurence Du-Thumm, Long Pan
An aqueous suspension of nanogibbsite was synthesized via the titration of aluminum aqua acid [Al(H2O)6](3+) with L-arginine to pH 4.6. Since the hydrolysis of aqueous aluminum salts is known to produce a wide array of products with a wide range of size distributions, a variety of state-of-the-art instruments (i.e., (27)Al/(1)H NMR, FTIR, ICP-OES, TEM-EDX, XPS, XRD, and BET) were used to characterize the synthesis products and identification of byproducts. The product, which was comprised of nanoparticles (10-30 nm), was isolated using gel permeation chromatography (GPC) column technique...
May 11, 2017: Journal of Visualized Experiments: JoVE
https://www.readbyqxmd.com/read/28518123/expression-of-exogenous-cytokine-in-patient-derived-xenografts-via-injection-with-a-cytokine-transduced-stromal-cell-line
#5
Jacqueline S Coats, Ineavely Baez, Cornelia Stoian, Terry-Ann M Milford, Xiaobing Zhang, Olivia L Francis, Ruijun Su, Kimberly J Payne
Patient-derived xenograft (PDX) mice are produced by transplanting human cells into immune deficient mice. These models are an important tool for studying the mechanisms of normal and malignant hematopoiesis and are the gold standard for identifying effective chemotherapies for many malignancies. PDX models are possible because many of the mouse cytokines also act on human cells. However, this is not the case for all cytokines, including many that are critical for studying normal and malignant hematopoiesis in human cells...
May 10, 2017: Journal of Visualized Experiments: JoVE
https://www.readbyqxmd.com/read/28518118/harnessing-the-dna-dye-triggered-side-population-phenotype-to-detect-and-purify-cancer-stem-cells-from-biological-samples
#6
Maximilian Boesch, Elisabeth Hoflehner, Dominik Wolf, Guenther Gastl, Sieghart Sopper
Cancer is a stem cell-driven disease and eradication of these cells has become a major therapeutic goal. Deciphering vulnerabilities of Cancer Stem Cells (CSCs) and identifying suitable molecular targets relies on methods that allow their specific discrimination in heterogeneous samples such as cell lines and ex vivo tumor tissue. Flow cytometry/FACS is a powerful technology to multi-parametrically dissect biological samples at the single cell level and is to date the method of choice to recover live cells for downstream analyses...
May 10, 2017: Journal of Visualized Experiments: JoVE
https://www.readbyqxmd.com/read/28518109/assessment-of-neuronal-viability-using-fluorescein-diacetate-propidium-iodide-double-staining-in-cerebellar-granule-neuron-culture
#7
Lin Jiajia, Mak Shinghung, Zheng Jiacheng, Wang Jialing, Xu Dilin, Hu Shengquan, Zhang Zaijun, Wang Qinwen, Han Yifan, Cui Wei
Primary cultured Cerebellar Granule Neurons (CGNs) have been widely used as an in vitro model in neuroscience and neuropharmacology research. However, the co-existence of glial cells and neurons in CGN culture might lead to biases in the accurate assessment of neuronal viability. Fluorescein diacetate (FDA) and Propidium Iodide (PI) double staining has been used to measure cell viability by simultaneously evaluating the viable and dead cells. We used FDA-PI double staining to improve the sensitivities of the colorimetric assays and to evaluate neuronal viability in CGNs...
May 10, 2017: Journal of Visualized Experiments: JoVE
https://www.readbyqxmd.com/read/28518101/using-wavelet-entropy-to-demonstrate-how-mindfulness-practice-increases-coordination-between-irregular-cerebral-and-cardiac-activities
#8
Hin Hung Sik, Junling Gao, Jicong Fan, Bonnie Wai Yan Wu, Hang Kin Leung, Yeung Sam Hung
In both the East and West, traditional teachings say that the mind and heart are somehow closely correlated, especially during spiritual practice. One difficulty in proving this objectively is that the natures of brain and heart activities are quite different. In this paper, we propose a methodology that uses wavelet entropy to measure the chaotic levels of both electroencephalogram (EEG) and electrocardiogram (ECG) data and show how this may be used to explore the potential coordination between the mind and heart under different experimental conditions...
May 10, 2017: Journal of Visualized Experiments: JoVE
https://www.readbyqxmd.com/read/28518084/subcutaneous-trigeminal-nerve-field-stimulation-for-refractory-facial-pain
#9
Martin Jakobs, Sigrid Schuh-Hofer, Andreas Unterberg, Rezvan Ahmadi
Chronic or neuropathic trigeminal facial pain can be challenging to treat. Neurosurgical procedures should be applied when conservative treatment fails. Neuromodulation techniques for chronic facial pain include deep brain stimulation and motor cortex stimulation, which are complex to perform. Subcutaneous nerve field stimulation is certified for chronic back pain and is the least invasive form of neuromodulation. We applied this technique to treat chronic and neuropathic trigeminal pain as an individual therapy concept...
May 10, 2017: Journal of Visualized Experiments: JoVE
https://www.readbyqxmd.com/read/28518129/cardiac-muscle-cell-based-actuator-and-self-stabilizing-biorobot-part-2
#10
Neerajha Nagarajan, Merrel T Holley, Christian Danielson, Kidong Park, Pinar Zorlutuna
In recent years, hybrid devices that consist of a living cell or tissue component integrated with a synthetic mechanical backbone have been developed. These devices, called biorobots, are powered solely by the force generated from the contractile activity of the living component and, due to their many inherent advantages, could be an alternative to conventional fully artificial robots. Here, we describe the methods to seed and characterize a biological actuator and a biorobot that was designed, fabricated, and functionalized in the first part of this two-part article...
May 9, 2017: Journal of Visualized Experiments: JoVE
https://www.readbyqxmd.com/read/28518075/leveraging-cyverse-resources-for-de-novo-comparative-transcriptomics-of-underserved-non-model-organisms
#11
Blake L Joyce, Asher K Haug-Baltzell, Jonathan P Hulvey, Fiona McCarthy, Upendra Kumar Devisetty, Eric Lyons
This workflow allows novice researchers to leverage advanced computational resources such as cloud computing to carry out pairwise comparative transcriptomics. It also serves as a primer for biologists to develop data scientist computational skills, e.g. executing bash commands, visualization and management of large data sets. All command line code and further explanations of each command or step can be found on the wiki (https://wiki.cyverse.org/wiki/x/dgGtAQ). The Discovery Environment and Atmosphere platforms are connected together through the CyVerse Data Store...
May 9, 2017: Journal of Visualized Experiments: JoVE
https://www.readbyqxmd.com/read/28518074/eliciting-and-analyzing-male-mouse-ultrasonic-vocalization-usv-songs
#12
Jonathan Chabout, Joshua Jones-Macopson, Erich D Jarvis
Mice produce ultrasonic vocalizations (USVs) in a variety of social contexts throughout development and adulthood. These USVs are used for mother-pup retrieval(1), juvenile interactions(2), opposite and same sex interactions(3)(,)(4)(,)(5), and territorial interactions(6). For decades, the USVs have been used by investigators as proxies to study neuropsychiatric and developmental or behavioral disorders(7)(,)(8)(,)(9), and more recently to understand mechanisms and evolution of vocal communication among vertebrates(10)...
May 9, 2017: Journal of Visualized Experiments: JoVE
https://www.readbyqxmd.com/read/28518128/an-in-vitro-caseum-binding-assay-that-predicts-drug-penetration-in-tuberculosis-lesions
#13
Jansy P Sarathy, Hsin-Pin Ho Liang, Danielle Weiner, Jacqueline Gonzales, Laura E Via, Véronique Dartois
The eradication of tuberculosis disease requires drug regimens that can penetrate the multiple layers of complex pulmonary lesions. Drug distribution in the caseous cores of cavities and lesions is especially crucial because they harbor subpopulations of drug-tolerant bacteria also commonly referred to as persisters. Existing methods for the measurement of drug penetration in tuberculosis lesions involve costly and time-consuming in vivo pharmacokinetic studies coupled to bioanalytical or imaging techniques...
May 8, 2017: Journal of Visualized Experiments: JoVE
https://www.readbyqxmd.com/read/28518116/the-wincf-model-an-inexpensive-and-tractable-microcosm-of-a-mucus-plugged-bronchiole-to-study-the-microbiology-of-lung-infections
#14
William J Comstock, Edwin Huh, Reiley Weekes, Connor Watson, Tianyang Xu, Pieter C Dorrestein, Robert A Quinn
Many chronic airway diseases result in mucus plugging of the airways. Lungs of an individual with cystic fibrosis are an exemplary case where their mucus-plugged bronchioles create a favorable habitat for microbial colonization. Various pathogens thrive in this environment interacting with each other and driving many of the symptoms associated with CF disease. Like any microbial community, the chemical conditions of their habitat have a significant impact on the community structure and dynamics. For example, different microorganisms thrive in differing levels of oxygen or other solute concentrations...
May 8, 2017: Journal of Visualized Experiments: JoVE
https://www.readbyqxmd.com/read/28518080/rearing-ixodes-scapularis-the-black-legged-tick-feeding-immature-stages-on-mice
#15
Andrew B Nuss, Manoj G Mathew, Monika Gulia-Nuss
Ixodes scapularis, the vector of Lyme disease, is one of the most important disease vectors in the eastern and Midwestern United States. This species is a three host tick that requires a blood meal from a vertebrate host for each development stage, and the adult females require a blood meal for reproduction. Larval ticks attach to their host for 3 - 5 days for feeding and drop off the host when fully engorged. This dependency on several different hosts and the lengthy attachment time for engorgement complicates tick rearing in the laboratory setting...
May 8, 2017: Journal of Visualized Experiments: JoVE
https://www.readbyqxmd.com/read/28518078/non-invasive-in-vivo-fluorescence-optical-imaging-of-inflammatory-mmp-activity-using-an-activatable-fluorescent-imaging-agent
#16
Johannes Schwenck, Florian C Maier, Manfred Kneilling, Stefan Wiehr, Kerstin Fuchs
This paper describes a non-invasive method for imaging matrix metalloproteinases (MMP)-activity by an activatable fluorescent probe, via in vivo fluorescence optical imaging (OI), in two different mouse models of inflammation: a rheumatoid arthritis (RA) and a contact hypersensitivity reaction (CHR) model. Light with a wavelength in the near infrared (NIR) window (650 - 950 nm) allows a deeper tissue penetration and minimal signal absorption compared to wavelengths below 650 nm. The major advantages using fluorescence OI is that it is cheap, fast and easy to implement in different animal models...
May 8, 2017: Journal of Visualized Experiments: JoVE
https://www.readbyqxmd.com/read/28518117/simultaneous-evaluation-of-cerebral-hemodynamics-and-light-scattering-properties-of-the-in-vivo-rat-brain-using-multispectral-diffuse-reflectance-imaging
#17
Izumi Nishidate, Afrina Mustari, Satoko Kawauchi, Shunichi Sato, Manabu Sato
The simultaneous evaluation of cerebral hemodynamics and the light scattering properties of in vivo rat brain tissue is demonstrated using a conventional multispectral diffuse reflectance imaging system. This system is constructed from a broadband white light source, a motorized filter wheel with a set of narrowband interference filters, a light guide, a collecting lens, a video zoom lens, and a monochromatic charged-coupled device (CCD) camera. An ellipsoidal cranial window is made in the skull bone of a rat under isoflurane anesthesia to capture in vivo multispectral diffuse reflectance images of the cortical surface...
May 7, 2017: Journal of Visualized Experiments: JoVE
https://www.readbyqxmd.com/read/28518110/external-excitation-of-neurons-using-electric-and-magnetic-fields-in-one-and-two-dimensional-cultures
#18
Shani Stern, Assaf Rotem, Yuri Burnishev, Eyal Weinreb, Elisha Moses
A neuron will fire an action potential when its membrane potential exceeds a certain threshold. In typical activity of the brain, this occurs as a result of chemical inputs to its synapses. However, neurons can also be excited by an imposed electric field. In particular, recent clinical applications activate neurons by creating an electric field externally. It is therefore of interest to investigate how the neuron responds to the external field and what causes the action potential. Fortunately, precise and controlled application of an external electric field is possible for embryonic neuronal cells that are excised, dissociated and grown in cultures...
May 7, 2017: Journal of Visualized Experiments: JoVE
https://www.readbyqxmd.com/read/28518107/a-large-lateral-craniotomy-procedure-for-mesoscale-wide-field-optical-imaging-of-brain-activity
#19
Michael Kyweriga, Jianjun Sun, Sunny Wang, Richard Kline, Majid H Mohajerani
The craniotomy is a commonly performed procedure to expose the brain for in vivo experiments. In mouse research, most labs utilize a small craniotomy, typically 3 mm x 3 mm. This protocol introduces a method for creating a substantially larger 7 mm x 6 mm cranial window exposing most of a cerebral hemisphere over the mouse temporal and parietal cortices (e.g., bregma 2.5 - 4.5 mm, lateral 0 - 6 mm). To perform this surgery, the head must be tilted approximately 30° and much of the temporal muscle must be retracted...
May 7, 2017: Journal of Visualized Experiments: JoVE
https://www.readbyqxmd.com/read/28518091/long-term-live-imaging-of-drosophila-eye-disc
#20
Chia-Kang Tsao, Hui-Yu Ku, Y Henry Sun
Live imaging provides the ability to continuously track dynamic cellular and developmental processes in real time. Drosophila larval imaginal discs have been used to study many biological processes, such as cell proliferation, differentiation, growth, migration, apoptosis, competition, cell-cell signaling, and compartmental boundary formation. However, methods for the long-term ex vivo culture and live imaging of the imaginal discs have not been satisfactory, despite many efforts. Recently, we developed a method for the long-term ex vivo culture and live imaging of imaginal discs for up to 18 h...
May 6, 2017: Journal of Visualized Experiments: JoVE
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