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Electroporation

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https://www.readbyqxmd.com/read/28935710/neural-specific-deletion-of-the-focal-adhesion-adaptor-protein-paxillin-slows-migration-speed-and-delays-cortical-layer-formation
#1
Mamunur Rashid, Judson Belmont, David Carpenter, Christopher E Turner, Eric C Olson
Paxillin and Hic-5 are homologous focal adhesion adaptor proteins that coordinate cytoskeletal rearrangements in response to integrin-signaling, but their role(s) in cortical development are unknown. Here, we find that Hic-5 deficient mice are postnatal viable with normal cortical layering. Mice with a neural-specific deletion of paxillin are also postnatal viable, but show evidence of a cortical neuron migration delay that is evident pre and perinatally, but is not detected at postnatal day 35 (P35). This phenotype is not modified by Hic-5 deficiency (double knockout)...
September 21, 2017: Development
https://www.readbyqxmd.com/read/28935594/self-replicating-rna-vaccine-functionality-modulated-by-fine-tuning-of-polyplex-delivery-vehicle-structure
#2
Thomas Démoulins, Thomas Ebensen, Kai Schulze, Pavlos C Englezou, Panagiota Milona, Maria Pelliccia, Carlos A Guzmán, Nicolas Ruggli, Kenneth C McCullough
The major limitations with large and complex self-amplifying RNA vaccines (RepRNA) are RNase-sensitivity and inefficient translation in dendritic cells (DCs). Condensing RepRNA with polyethylenimine (PEI) gave positive in vitro readouts, but was largely inferior to virus-like replicon particles (VRP) or direct electroporation. In the present study, we improved such polyplex formulation and determined that fine-tuning of the polyplex structure is essential for ensuring efficacious translation. Thereby, three parameters dominate: (i) PEI molecular weight (MW); (ii) RepRNA:PEI (weight:weight) ratio; and (iii) inclusion of cell penetrating peptides (CPPs)...
September 18, 2017: Journal of Controlled Release: Official Journal of the Controlled Release Society
https://www.readbyqxmd.com/read/28934689/dynamical-modeling-of-tissue-electroporation
#3
Damien Voyer, Aude Silve, Lluis M Mir, Riccardo Scorretti, Clair Poignard
In this paper, we propose a new dynamical model of tissue electroporation. The model is based on equivalent circuit approach at the tissue. Considering two current densities from cells and extracellular matrix, we identify the macroscopic homogenised contribution of the cell membranes. Our approach makes it possible to define a macroscopic homogenised electric field and a macroscopic homogenised transmembrane potential. This provides a direct link between the cell scale electroporation models and the tissue models...
September 5, 2017: Bioelectrochemistry
https://www.readbyqxmd.com/read/28932622/high-throughput-nuclear-delivery-and-rapid-expression-of-dna-via-mechanical-and-electrical-cell-membrane-disruption
#4
Xiaoyun Ding, Martin Stewart, Armon Sharei, James C Weaver, Robert S Langer, Klavs F Jensen
Nuclear transfection of DNA into mammalian cells is challenging yet critical for many biological and medical studies. Here, by combining cell squeezing and electric-field-driven transport in a device that integrates microfluidic channels with constrictions and microelectrodes, we demonstrate nuclear delivery of plasmid DNA within 1 hour after treatment, the most rapid DNA expression in a high-throughput setting (up to millions of cells per minute per device). Passing cells at high speed through microfluidic constrictions smaller than the cell diameter mechanically disrupts the cell membrane, allowing a subsequent electric field to further disrupt the nuclear envelope and drive DNA molecules into the cytoplasm and nucleus...
2017: Nature biomedical engineering
https://www.readbyqxmd.com/read/28932093/postoperative-inflammation-as-a-possible-cause-of-portal-vein-thrombosis-after-irreversible-electroporation-for-locally-advanced-pancreatic-cancer
#5
Jun-Jun Su, Ming Su, Kai Xu, Peng-Fei Wang, Li Yan, Shi-Chun Lu, Wan-Qing Gu, Yong-Liang Chen
Portal vein thrombosis (PVT) is a rare but serious postoperative complication associated with irreversible electroporation (IRE). We report a case of postoperative PVT in a 54-year-old woman who underwent IRE for locally advanced pancreatic cancer. Drain removal and discharge of the patient from the hospital were scheduled on postoperative day (POD) 7; however, a magnetic resonance imaging scan revealed the presence of PVT. We suspected postoperative inflammation in the pancreas as the main cause of PVT. However, the patient did not undergo any medical treatment because she did not have any clinical symptoms, and she was discharged on POD 8...
August 28, 2017: World Journal of Gastroenterology: WJG
https://www.readbyqxmd.com/read/28931520/dna-priming-increases-frequency-of-t-cell-responses-to-a-vsv-hiv-vaccine-with-specific-enhancement-of-cd8-t-cell-responses-by-il-12-pdna
#6
Shuying S Li, Nidhi K Kochar, Marnie Elizaga, Christine M Hay, Gregory J Wilson, Kristen W Cohen, Stephen C De Rosa, Rong Xu, Ayuko Ota-Setlik, Daryl Morris, Greg Finak, Mary Allen, Hong-Van Tieu, Ian Frank, Magdalena E Sobieszczyk, Drew Hannaman, Raphael Gottardo, Peter B Gilbert, Georgia D Tomaras, Lawrence Corey, David K Clarke, Michael A Egan, John H Eldridge, M Juliana McElrath, Nicole Frahm
HVTN 087 assessed the effect of increasing doses of pIL-12 adjuvant on the immunogenicity of a HIV-1 multi-antigen (MAG) DNA vaccine delivered by electroporation and boosted with an attenuated VSV-Gag vaccine. We randomized 100 healthy adults to receive placebo, or 3mg HIV-MAG (gag/pol, env, nef/tat/vif) DNA vaccine co-administered with pIL-12 at 0μg, 250μg, 1,000μg or 1,500μg intramuscularly by electroporation at 0, 1 and 3 months followed by intramuscular inoculation with 3.4×10(7) PFU VSV-Gag vaccine at 6 months...
September 20, 2017: Clinical and Vaccine Immunology: CVI
https://www.readbyqxmd.com/read/28929209/upper-urinary-tract-effects-after-irreversible-electroporation-ire-of-human-localised-renal-cell-carcinoma-rcc-in-the-irene-pilot-phase-2a-ablate-and-resect-study
#7
J J Wendler, M Pech, J Köllermann, B Friebe, S Siedentopf, S Blaschke, D Schindele, M Porsch, D Baumunk, J Jürgens, F Fischbach, J Ricke, M Schostak, M Böhm, U B Liehr
PURPOSE: Irreversible electroporation (IRE) is a new potential ablation modality for small renal masses. Animal experiments have shown preservation of the urine-collecting system (UCS). The purpose of this clinical study was to perform the first evaluation and comparison of IRE's effects on the renal UCS by using urinary cytology, magnetic-resonance imaging, and resection histology in men after IRE of pT1a renal-cell carcinoma (RCC). METHODS: Seven patients with biopsy-proven RCC pT1a cN0cM0 underwent IRE in a phase 2a pilot ablate-and-resect study (IRENE trial)...
September 19, 2017: Cardiovascular and Interventional Radiology
https://www.readbyqxmd.com/read/28928022/the-role-of-jab1-a-putative-downstream-effector-of-the-neurotrophic-cytokine-macrophage-migration-inhibitory-factor-mif-in-zebrafish-inner-ear-hair-cell-development
#8
REVIEW
Loren J Weber, Hannah K Marcy, Yu-Chi Shen, Sarah E Tomkovich, Kristina M Brooks, Kelly E Hilk, Kate F Barald
Macrophage migration inhibitory factor (MIF) is a neurotrophic cytokine essential for inner ear hair cell (HC) development and statoacoustic ganglion (SAG) neurite outgrowth, and SAG survival in mouse, chick and zebrafish. Another neurotrophic cytokine, Monocyte chemoattractant protein 1 (MCP1) is known to synergize with MIF; but MCP1 alone is insufficient to support mouse/chick SAG neurite outgrowth or neuronal survival. Because of the relatively short time over which the zebrafish inner ear develops (~30h), the living zebrafish embryo is an ideal system to examine mif and mcp1 cytokine pathways and interactions...
September 16, 2017: Experimental Neurology
https://www.readbyqxmd.com/read/28927918/the-safety-of-irreversible-electroporation-on-nerves-adjacent-to-treated-tumors
#9
Xiaomei Luo, Zilin Qin, Huimin Tao, Jian Shi, Gang Fang, Zhonghai Li, Xulong Zhou, Jibing Chen, Kecheng Xu, Jianying Zeng, Lizhi Niu
OBJECTIVES: To evaluate the safety of irreversible electroporation (IRE) on the sciatic nerve after IRE ablation of adjacent tumor. METHODS: and analysis In this study, VX2 tumors were implanted adjacent to the sciatic nerves in 26 New Zealand white rabbits. The rabbits were divided into three groups 10 days after implantation, with two treatment groups of ten rabbits each and a control group of six rabbits. In one of the treatment groups (IRE-S group), the rabbits underwent ablation with a single IRE application, and in the other group (IRE-D group) the rabbits underwent two IRE applications...
September 16, 2017: World Neurosurgery
https://www.readbyqxmd.com/read/28927382/a-rapid-seamless-method-for-gene-knockout-in-pseudomonas-aeruginosa
#10
Weiliang Huang, Angela Wilks
BACKGROUND: Pseudomonas aeruginosa is a model organism for the study of quorum sensing, biofilm formation, and also leading cause of nosocomial infections in immune compromised patients. As such P. aeruginosa is one of the most well studied organisms in terms of its genetics. However, the construction of gene deletions and replacements in Pseudomonas aeruginosa is relatively time-consuming, requiring multiple steps including suicide vector construction, conjugation, inactivation with insertion of antibiotic resistance cassettes and allelic exchange...
September 19, 2017: BMC Microbiology
https://www.readbyqxmd.com/read/28924200/controllable-moderate-heating-enhances-the-therapeutic-efficacy-of-irreversible-electroporation-for-pancreatic-cancer
#11
Chelsea M Edelblute, James Hornef, Niculina I Burcus, Thomas Norman, Stephen J Beebe, Karl Schoenbach, Richard Heller, Chunqi Jiang, Siqi Guo
Irreversible electroporation (IRE) as a non-thermal tumor ablation technology has been studied for the treatment of pancreatic carcinoma and has shown a significant survival benefit. We discovered that moderate heating (MH) at 43 °C for 1-2 minutes significantly enhanced ex vivo IRE tumor ablation of Pan02 cells by 5.67-fold at 750 V/cm and by 1.67-fold at 1500 V/cm. This amount of heating alone did not cause cell death. An integrated IRE system with controllable laser heating and tumor impedance monitoring was developed to treat mouse ectopic pancreatic cancer...
September 18, 2017: Scientific Reports
https://www.readbyqxmd.com/read/28922628/high-frequency-electroporation-efficiency-is-under-control-of-membrane-capacitive-charging-and-voltage-potential-relaxation
#12
Vitalij Novickij, Paulius Ruzgys, Audrius Grainys, Saulius Šatkauskas
The study presents the proof of concept for a possibility to achieve a better electroporation in the MHz pulse repetition frequency (PRF) region compared to the conventional low frequency protocols. The 200ns×10 pulses bursts of 10-14kV/cm have been used to permeabilize Chinese hamster ovary (CHO) cells in a wide range (1Hz-1MHz) of PRF. The permeabilization efficiency was evaluated using fluorescent dye assay (propidium iodide) and flow cytometry. It was determined that a threshold PRF exists when the relaxation of the cell transmembrane potential is longer than the delay between the consequent pulses, which results in accumulation of the charge on the membrane...
September 14, 2017: Bioelectrochemistry
https://www.readbyqxmd.com/read/28922426/a-dna-vaccine-for-crimean-congo-hemorrhagic-fever-protects-against-disease-and-death-in-two-lethal-mouse-models
#13
Aura R Garrison, Charles J Shoemaker, Joseph W Golden, Collin J Fitzpatrick, John J Suschak, Michelle J Richards, Catherine V Badger, Carolyn M Six, Jacqueline D Martin, Drew Hannaman, Marko Zivcec, Eric Bergeron, Jeffrey W Koehler, Connie S Schmaljohn
Crimean-Congo hemorrhagic fever virus (CCHFV) is a tick-borne virus capable of causing a severe hemorrhagic fever disease in humans. There are currently no licensed vaccines to prevent CCHFV-associated disease. We developed a DNA vaccine expressing the M-segment glycoprotein precursor gene of CCHFV and assessed its immunogenicity and protective efficacy in two lethal mouse models of disease: type I interferon receptor knockout (IFNAR-/-) mice; and a novel transiently immune suppressed (IS) mouse model. Vaccination of mice by muscle electroporation of the M-segment DNA vaccine elicited strong antigen-specific humoral immune responses with neutralizing titers after three vaccinations in both IFNAR-/- and IS mouse models...
September 18, 2017: PLoS Neglected Tropical Diseases
https://www.readbyqxmd.com/read/28919588/poly-i-c-delivery-into-j774-1-and-raw264-7-macrophages-induces-rapid-cell-death
#14
Cheah Wen Yapp, Amal Widaad Mohaimin, Adi Idris
BACKGROUND: Cytosolic double-stranded RNA (dsRNA) is an important 'molecular signature' for the detection of intracellular viral infections. Although intracellular dsRNA is a known potent inducer of apoptosis, the optimal time and dose for the onset of dsRNA-mediated apoptosis have not been studied in detail. OBJECTIVE: To perform an accurate temporal assessment of the cell death responses in dsRNA-dependent cytotoxicity. METHODS: Poly I:C (PIC), a synthetic dsRNA molecule was delivered intracellularly into J774...
September 2017: Iranian Journal of Immunology: IJI
https://www.readbyqxmd.com/read/28918039/crispr-cas9-mediated-knockin-application-in-cell-therapy-a-non-viral-procedure-for-bystander-treatment-of-glioma-in-mice
#15
Oscar Meca-Cortés, Marta Guerra-Rebollo, Cristina Garrido, Salvador Borrós, Nuria Rubio, Jeronimo Blanco
The use of non-viral procedures, together with CRISPR/Cas9 genome-editing technology, allows the insertion of single-copy therapeutic genes at pre-determined genomic sites, overcoming safety limitations resulting from random gene insertions of viral vectors with potential for genome damage. In this study, we demonstrate that combination of non-viral gene delivery and CRISPR/Cas9-mediated knockin via homology-directed repair can replace the use of viral vectors for the generation of genetically modified therapeutic cells...
September 15, 2017: Molecular Therapy. Nucleic Acids
https://www.readbyqxmd.com/read/28917658/multiple-events-of-gene-manipulation-via-in-pouch-electroporation-in-a-marsupial-model-of-mammalian-forebrain-development
#16
Annalisa Paolino, Laura R Fenlon, Peter Kozulin, Linda J Richards, Rodrigo Suárez
BACKGROUND: The technique of in utero electroporation has been widely used in eutherians, such as mice and rats, to investigate brain development by selectively manipulating gene expression in specific neuronal populations. A major challenge, however, is that surgery is required to access the embryos, affecting animal survival and limiting the number of times it can be performed within the same litter. New method: Marsupials are born at an early stage of brain development as compared to eutherians...
September 13, 2017: Journal of Neuroscience Methods
https://www.readbyqxmd.com/read/28913821/erratum-to-foreword-to-the-special-issue-electroporation-for-biomedical-applications
#17
Günther Zeck, Damijan Miklavčič
No abstract text is available yet for this article.
September 15, 2017: Medical & Biological Engineering & Computing
https://www.readbyqxmd.com/read/28913803/doubled-haploid-transgenic-wheat-lines-by-microspore-transformation
#18
Sachin Rustgi, Nii O Ankrah, Rhoda A T Brew-Appiah, Yue Sun, Weiguo Liu, Diter von Wettstein
Microspores are preferred explant choice for genetic transformation, as their use shortens the duration of obtaining homozygous transformants. All established gene-delivery methods of particle bombardment, electroporation, and cocultivation with Agrobacterium tumefaciens were optimized on androgenic microspores or derived tissues. In the biolistic gene delivery method 35-40 days old haploid microspore embryoids were used for genetic transformation, whereas freshly isolated androgenic microspores were used for genetic transformation in the electroporation and Agrobacterium cocultivation-based methods...
2017: Methods in Molecular Biology
https://www.readbyqxmd.com/read/28912684/distinct-activities-of-tfap2a-and-tfap2b-in-the-specification-of-gabaergic-interneurons-in-the-developing-cerebellum
#19
Norliyana Zainolabidin, Sandhya P Kamath, Ayesha R Thanawalla, Albert I Chen
GABAergic inhibitory neurons in the cerebellum are subdivided into Purkinje cells and distinct subtypes of interneurons from the same pool of progenitors, but the determinants of this diversification process are not well defined. To explore the transcriptional regulation of the development of cerebellar inhibitory neurons, we examined the role of Tfap2A and Tfap2B in the specification of GABAergic neuronal subtypes in mice. We show that Tfap2A and Tfap2B are expressed in inhibitory precursors during embryonic development and that their expression persists into adulthood...
2017: Frontiers in Molecular Neuroscience
https://www.readbyqxmd.com/read/28901954/avoiding-nerve-stimulation-in-irreversible-electroporation-a-numerical-modeling-study
#20
Borja Mercadal, Christopher B Arena, Rafael V Davalos, Antoni Ivorra
Electroporation based treatments consist in applying one or multiple high voltage pulses to the tissues to be treated. As an undesired side effect, these pulses cause electrical stimulation of excitable tissues such as nerves and muscles. This increases the complexity of the treatments and may pose a risk to the patient. To minimize electrical stimulation during electroporation based treatments, it has been proposed to replace the commonly used monopolar pulses by bursts of short bipolar pulses. In the present study, we have numerically analyzed the rationale for such approach...
September 13, 2017: Physics in Medicine and Biology
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