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Electroporation

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https://www.readbyqxmd.com/read/28448060/small-rna-transfection-in-primary-human-th17-cells-by-next-generation-electroporation
#1
Misty M Montoya, K Mark Ansel
CD4(+) T cells can differentiate into several subsets of effector T helper cells depending on the surrounding cytokine milieu. Th17 cells can be generated from naïve CD4(+) T cells in vitro by activating them in the presence of the polarizing cytokines IL-1β, IL-6, IL-23, and TGFβ. Th17 cells orchestrate immunity against extracellular bacteria and fungi, but their aberrant activity has also been associated with several autoimmune and inflammatory diseases. Th17 cells are identified by the chemokine receptor CCR6 and defined by their master transcription factor, RORγt, and characteristic effector cytokine, IL-17A...
April 13, 2017: Journal of Visualized Experiments: JoVE
https://www.readbyqxmd.com/read/28445365/improvement-of-flap-necrosis-in-a-rat-random-skin-flap-model-by-in-vivo-electroporation-mediated-hgf-gene-transfer
#2
S Morteza Seyed Jafari, Maziar Shafighi, Helmut Beltraminelli, Thomas Geiser, Robert E Hunger, Amiq Gazdhar
BACKGROUND: Despite great understanding of underlying mechanisms for flap necrosis and advances in surgical techniques, flap necrosis remains a critical issue. In the present study, the authors investigated the efficacy of electroporation-mediated hepatocyte growth factor (HGF) gene delivery to random dorsal skin flaps (McFarlane) to accelerate wound healing and reduce flap necrosis. METHODS: Fifteen male Wistar rats (290 to 320 g) were divided randomly into three groups...
May 2017: Plastic and Reconstructive Surgery
https://www.readbyqxmd.com/read/28445252/irreversible-electroporation-in-primary-and-metastatic-hepatic-malignancies-a-review
#3
Tianchu Lyu, Xifu Wang, Zhanliang Su, Junjie Shangguan, Chong Sun, Matteo Figini, Jian Wang, Vahid Yaghmai, Andrew C Larson, Zhuoli Zhang
BACKGROUND: Liver cancer makes up a huge percentage of cancer mortality worldwide. Irreversible electroporation (IRE) is a relatively new minimally invasive nonthermal ablation technique for tumors that applies short pulses of high frequency electrical energy to irreversibly destabilize cell membrane to induce tumor cell apoptosis. METHODS: This review aims to investigate the studies regarding the use of IRE treatment in liver tumors and metastases to liver. We searched PubMed for all of IRE relevant English language articles published up to September 2016...
April 2017: Medicine (Baltimore)
https://www.readbyqxmd.com/read/28442784/the-rna-binding-protein-marf1-promotes-cortical-neurogenesis-through-its-rnase-activity-domain
#4
Yoshitaka Kanemitsu, Masashi Fujitani, Yuki Fujita, Suxiang Zhang, You-Qiang Su, Yukio Kawahara, Toshihide Yamashita
Cortical neurogenesis is a fundamental process of brain development that is spatiotemporally regulated by both intrinsic and extrinsic cues. Although recent evidence has highlighted the significance of transcription factors in cortical neurogenesis, little is known regarding the role of RNA-binding proteins (RBPs) in the post-transcriptional regulation of cortical neurogenesis. Here, we report that meiosis arrest female 1 (MARF1) is an RBP that is expressed during neuronal differentiation. Cortical neurons expressed the somatic form of MARF1 (sMARF1) but not the oocyte form (oMARF1)...
April 25, 2017: Scientific Reports
https://www.readbyqxmd.com/read/28441954/electrochemotherapy-efficacy-evaluation-for-treatment-of-locally-advanced-stage-iii-cutaneous-squamous-cell-carcinoma-a-22-cases-retrospective-analysis
#5
Gianluca Di Monta, Corrado Caracò, Ester Simeone, Antonio Maria Grimaldi, Ugo Marone, Massimiliano Di Marzo, Vito Vanella, Lucia Festino, Marco Palla, Stefano Mori, Nicola Mozzillo, Paolo Antonio Ascierto
BACKGROUND: Extensive squamous cell carcinoma has few therapeutic options. In such cases, electrochemotherapy involving electroporation combined with antineoplastic drug appears to be a new potential option and may be considered as an alternative treatment. The aim of this retrospective single-center study was to evaluate electrochemotherapy efficacy in treatment of locally advanced stage III squamous cell carcinoma, in which surgical procedures would have entailed wide tissue sacrifice...
April 26, 2017: Journal of Translational Medicine
https://www.readbyqxmd.com/read/28439465/single-exponential-decay-waveform-a-synergistic-combination-of-electroporation-and-electrolysis-e2-for-tissue-ablation
#6
Nina Klein, Enric Guenther, Paul Mikus, Michael K Stehling, Boris Rubinsky
BACKGROUND: Electrolytic ablation and electroporation based ablation are minimally invasive, non-thermal surgical technologies that employ electrical currents and electric fields to ablate undesirable cells in a volume of tissue. In this study, we explore the attributes of a new tissue ablation technology that simultaneously delivers a synergistic combination of electroporation and electrolysis (E2). METHOD: A new device that delivers a controlled dose of electroporation field and electrolysis currents in the form of a single exponential decay waveform (EDW) was applied to the pig liver, and the effect of various parameters on the extent of tissue ablation was examined with histology...
2017: PeerJ
https://www.readbyqxmd.com/read/28438941/new-macrolide-lincosamide-streptogramin-b-resistance-gene-erm-48-on-the-novel-plasmid-pjw2311-in-staphylococcus-xylosus
#7
Juliette R K Wipf, Matthew C Riley, Stephen A Kania, David A Bemis, Sabrina Andreis, Sybille Schwendener, Vincent Perreten
Whole-genome sequencing of Staphylococcus xylosus strain JW2311 from bovine mastitis milk identified the novel 49.3 kb macrolide-lincosamide-streptogramin B (MLSB) resistance plasmid pJW2311. It contained the macrolide resistance gene mph(C), the macrolide-streptogramin B resistance gene msr(A), and the new MLSB resistance gene erm (48) and could be transformed into S. aureus by electroporation. Functionality of erm (48) was demonstrated by cloning and expression in S. aureus.
April 24, 2017: Antimicrobial Agents and Chemotherapy
https://www.readbyqxmd.com/read/28438205/expression-of-the-clonostachys-rosea-lactonohydrolase-gene-by-lactobacillus-reuteri-to-increase-its-zearalenone-removing-ability
#8
Wen-Chun Yang, Tsui-Chun Hsu, Kuan-Chen Cheng, Je-Ruei Liu
BACKGROUND: Mycotoxins are secondary metabolites produced by filamentous fungi that can contaminate agricultural crops in the field as well as during harvest, transportation, processing, or storage. Zearalenone (ZEN), a non-steroidal estrogenic mycotoxin, produced by Fusarium species, has been shown to be associated with reproductive disorders in farm animals and to a lesser extent in hyperoestrogenic syndromes in humans. Thus, the decontamination of ZEN in foods and feeds is an important issue...
April 24, 2017: Microbial Cell Factories
https://www.readbyqxmd.com/read/28432032/neuronal-excitation-and-permeabilization-by-200-ns-pulsed-electric-field-an-optical-membrane-potential-study-with-fluovolt-dye
#9
Andrei G Pakhomov, Iurii Semenov, M Casciola, Shu Xiao
Electric field pulses of nano- and picosecond duration are a novel modality for neurostimulation, activation of Ca(2+) signaling, and tissue ablation. However it is not known how such brief pulses activate voltage-gated ion channels. We studied excitation and electroporation of hippocampal neurons by 200-ns pulsed electric field (nsPEF), by means of time-lapse imaging of the optical membrane potential (OMP) with FluoVolt dye. Electroporation abruptly shifted OMP to a more depolarized level, which was reached within <1ms...
April 18, 2017: Biochimica et Biophysica Acta
https://www.readbyqxmd.com/read/28430358/regulation-of-axon-arborization-pattern-in-the-developing-chick-ciliary-ganglion-possible-involvement-of-caspase-3
#10
Hidetaka Katow, Teppei Kanaya, Tomohisa Ogawa, Ryo Egawa, Hiromu Yawo
During a certain critical period in the development of the central and peripheral nervous systems, axonal branches and synapses are massively reorganized to form mature connections. In this process, neurons search their appropriate targets, expanding and/or retracting their axons. Recent work suggested that the caspase superfamily regulates the axon morphology. Here, we tested the hypothesis that caspase 3, which is one of the major executioners in apoptotic cell death, is involved in regulating the axon arborization...
April 21, 2017: Development, Growth & Differentiation
https://www.readbyqxmd.com/read/28417251/irreversible-electroporation-for-the-treatment-of-rabbit-vx2-breast-cancer
#11
Wenlong Zhang, Wei Chai, Jianying Zeng, Jibing Chen, Liqi Bi, Lizhi Niu
Irreversible electroporation (IRE) is considered predominantly as a non-thermal ablative technique that uses electrical fields to permeabilize cell membranes and lead to cell death. In this study, we evaluated the efficacy of IRE in the rabbit VX2 breast cancer model. Thirty-five female New Zealand white rabbits were inoculated against VX2 breast cancer cells. The rabbits were randomly divided into two groups: a control group of 15 rabbits and an IRE treatment group of 20 rabbits. Treatment and treatment outcome were evaluated by computerized tomography (CT) scan (plain or contrast enhanced), tumor growth curves and pathological examination including H&E, TUNEL, PCNA and CD31 staining...
June 2017: Biomedical Microdevices
https://www.readbyqxmd.com/read/28414960/calibration-of-on-chip-cell-electroporation-by-a-pseudo-volumetric-uptake-model
#12
Kisoo Kim, Won Gu Lee
Most conventional methods for assessing uptake of exogenous molecules and nanomaterials into cells use the projected two-dimensional (2D) area of uptake intensity into individual cells. However, since most cells have a three-dimensional (3D) spherical shape, volumetric uptake cannot be quantified accurately using 2D area analysis. Here, we present a method for calibrating the electroporative uptake intensity of small molecules by using a novel predictable spherical volume (PSV) model, which is more accurate and quantitative than previous methods...
April 10, 2017: Micron: the International Research and Review Journal for Microscopy
https://www.readbyqxmd.com/read/28413132/development-of-an-intradermal-dna-vaccine-delivery-strategy-to-achieve-single-dose-immunity-against-respiratory-syncytial-virus
#13
Trevor R F Smith, Katherine Schultheis, Matthew P Morrow, Kimberly A Kraynyak, Jay R McCoy, Kevin C Yim, Karuppiah Muthumani, Laurent Humeau, David B Weiner, Niranjan Y Sardesai, Kate E Broderick
Respiratory syncytial virus (RSV) is a massive medical burden in infants, children and the elderly worldwide, and an effective, safe RSV vaccine remains an unmet need. Here we assess a novel vaccination strategy based on the intradermal delivery of a SynCon® DNA-based vaccine encoding engineered RSV-F antigen using a surface electroporation device (SEP) to target epidermal cells, in clinically relevant experimental models. We demonstrate the ability of this strategy to elicit robust immune responses. Importantly we demonstrate complete resistance to pulmonary infection at a single low dose of vaccine in the cotton rat RSV/A challenge model...
April 13, 2017: Vaccine
https://www.readbyqxmd.com/read/28412668/electroporation-of-tissue-and-cells-a-three-equation-model-of-drug-delivery
#14
Finbar Argus, Bradley Boyd, S M Becker
The exposure of the cell membrane to electric pulses of sufficient intensity is known to result in an increased permeability due to the formation of microscopic pores. This is electroporation, and it has been implemented to increase the efficacy of targeted drug delivery. In this study we introduce a novel three-equation model of transport that is able to distinguish the drug uptake in reversibly electroporated cells from that in irreversibly electroporated cells. In order to relate the permeability increases and the cell survival to the local electric field, sigmoidal functions are fit to published experimental data...
April 7, 2017: Computers in Biology and Medicine
https://www.readbyqxmd.com/read/28412360/in-house-made-nucleofection-buffer-for-efficient-and-cost-effective-transfection-of-raw-264-7-macrophages
#15
Neeraja P Alamuru-Yellapragada, Bandish Kapadia, Kishore V L Parsa
Electroporation is the most widely employed method of gene transfer into macrophages which are hard to transfect. RAW 264.7 is a widely used cell line for studying macrophage responses. Electroporation of RAW 264.7 cells with commercial reagents although very efficient is expensive necessitating the development of cost effective alternatives. In this study, we have formulated an economical electroporation buffer for electroporation of RAW 264.7 cells compatible with commercial nucleofector apparatus. We observed that supplementation of membrane fusogenic agents such as Ficoll, PEG and membrane resealing agent, poloxamer P188, enhanced the transfection efficiency of macrophages to a level comparable to the commercially available solutions thereby providing us a cost effective solution for genetic manipulation of macrophages especially in large numbers...
April 12, 2017: Biochemical and Biophysical Research Communications
https://www.readbyqxmd.com/read/28411378/immune-responses-and-long-term-disease-recurrence-status-after-telomerase-based-dendritic-cell-immunotherapy-in-patients-with-acute-myeloid-leukemia
#16
Hanna J Khoury, Robert H Collins, William Blum, Patrick S Stiff, Laurence Elias, Jane S Lebkowski, Anita Reddy, Kevin P Nishimoto, Debasish Sen, Edward D Wirth, Casey C Case, John F DiPersio
BACKGROUND: Telomerase activity in leukemic blasts frequently is increased among patients with high-risk acute myeloid leukemia (AML). In the current study, the authors evaluated the feasibility, safety, immunogenicity, and therapeutic potential of human telomerase reverse transcriptase (hTERT)-expressing autologous dendritic cells (hTERT-DCs) in adult patients with AML. METHODS: hTERT-DCs were produced from patient-specific leukapheresis, electroporated with an mRNA-encoding hTERT and a lysosomal-targeting sequence, and cryopreserved...
April 14, 2017: Cancer
https://www.readbyqxmd.com/read/28404597/altered-mirna-29-expression-in-type-2-diabetes-influences-glucose-and-lipid-metabolism-in-skeletal-muscle
#17
Julie Massart, Rasmus J O Sjögren, Leonidas S Lundell, Jonathan M Mudry, Niclas Franck, Donal J O'Gorman, Brendan Egan, Juleen R Zierath, Anna Krook
MicroRNAs have emerged as important regulators of glucose and lipid metabolism in several tissues, however, their role in skeletal muscle remains poorly characterized. We determined the effects of the miR-29 family on glucose and lipid metabolism, and insulin responsiveness in skeletal muscle. We provide evidence that miR-29a and miR-29c are increased in skeletal muscle from type 2 diabetic patients and decreased following endurance training in healthy young men and in rats. In primary human skeletal muscle cells, inhibition and overexpression strategies demonstrate that miR-29a and miR-29c regulate glucose uptake and insulin-stimulated glucose metabolism...
April 12, 2017: Diabetes
https://www.readbyqxmd.com/read/28401477/a-recombinant-probiotic-lactobacillus-casei-expressing-the-clostridium-perfringens-%C3%AE-toxoid-as-an-orally-vaccine-candidate-against-gas-gangrene-and-necrotic-enteritis
#18
Mojtaba Alimolaei, Mehdi Golchin, Jalil Abshenas, Majid Ezatkhah, Mehrdad Shamsaddini Bafti
The alpha-toxin is one of the virulence factors of Clostridium perfringens for gas gangrene in humans and animals or necrotic enteritis in poultry. The C-terminal domain of this toxin ( cpa 247-370 ) was synthesized and cloned into pT1NX vector to construct the pT1NX-alpha plasmid. This surface-expressing plasmid was electroporated into Lactobacillus casei ATCC 393, generating the recombinant L. casei strain expressing alpha-toxoid (LC-α strain). Expression of this modified alpha-toxoid was confirmed by SDS-PAGE, immunoblotting, and direct immunofluorescence microscopy...
April 11, 2017: Probiotics and Antimicrobial Proteins
https://www.readbyqxmd.com/read/28398645/crispr-cas9-mediated-gene-knockout-in-the-mouse-brain-using-in-utero-electroporation
#19
Yohei Shinmyo, Hiroshi Kawasaki
This unit describes a highly efficient and rapid procedure for brain-specific disruption of genes in the developing mouse brain using pX330 plasmids expressing humanized Cas9 and single-guide RNAs (sgRNAs) against target genes. The pX330 plasmids are delivered into the rodent brain using in utero electroporation. Focusing on the Satb2 gene, which encodes an AT-rich DNA-binding transcription factor, we found that the introduction of pX330-Satb2 induced insertion/deletion (indel) mutations near the predicted cleavage site in the Satb2 gene, resulting in a dramatic reduction of Satb2 expression in post-mitotic neurons...
April 10, 2017: Current Protocols in Neuroscience
https://www.readbyqxmd.com/read/28398641/time-lapse-imaging-and-cell-tracking-of-migrating-cells-in-slices-and-flattened-telencephalic-vesicles
#20
Verónica Murcia-Belmonte, Giovanna Expósito, Eloísa Herrera
Neuronal migration is a vital process needed for subsequent assembly and function of neural circuitry during embryonic development. The vast majority of neural progenitors are generated far from their final destination and need to migrate considerable distances to reach their specific cortical layer. Innovations in cell culture techniques and fluorescence microscopy now facilitate the direct visualization of cell movements during cortical development. Here, a detailed protocol to record and analyze a particular type of early migrating neurons, the Cajal Retzius Cells, during the development of the telencephalic vesicles in mammals is described...
April 10, 2017: Current Protocols in Neuroscience
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