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https://www.readbyqxmd.com/read/29026094/electropermeabilization-of-inner-and-outer-cell-membranes-with-microsecond-pulsed-electric-fields-quantitative-study-with-calcium-ions
#1
Hanna Hanna, Agnese Denzi, Micaela Liberti, Franck M André, Lluis M Mir
Microsecond pulsed electric fields (μsPEF) permeabilize the plasma membrane (PM) and are widely used in research, medicine and biotechnology. For internal membranes permeabilization, nanosecond pulsed electric fields (nsPEF) are applied but this technology is complex to use. Here we report that the endoplasmic reticulum (ER) membrane can also be electropermeabilized by one 100 µs pulse without affecting the cell viability. Indeed, using Ca(2+) as a permeabilization marker, we observed cytosolic Ca(2+) peaks in two different cell types after one 100 µs pulse in a medium without Ca(2+)...
October 12, 2017: Scientific Reports
https://www.readbyqxmd.com/read/28982976/activation-of-the-phospholipid-scramblase-tmem16f-by-nanosecond-pulsed-electric-field-nspef-facilitates-its-diverse-cytophysiological-effects
#2
Claudia Muratori, Andrei G Pakhomov, Elena Gianulis, Jade Meads, Maura Casciola, Peter A Mollica, Olga N Pakhomova
Nanosecond pulsed electric fields (nsPEFs) are emerging as a novel modality for cell stimulation and tissue ablation. However, the downstream protein effectors responsible for nsPEF bioeffects remain to be established. Here we demonstrate that nsPEFs activate TMEM16F, (or anoctamin 6), a protein functioning as a Ca(2+)-dependent phospholipid scramblase and Ca(2+)-activated chloride channel. Using confocal microscopy and patch-clamp recording, we investigated the relevance of TMEM16F activation for several bioeffects triggered by nsPEF, including phosphatidylserine (PS) externalization, nanopore-conducted currents, membrane blebbing, and cell death...
October 5, 2017: Journal of Biological Chemistry
https://www.readbyqxmd.com/read/28955986/nspef-induced-pip2-depletion-plc-activity-and-actin-cytoskeletal-cortex-remodeling-are-responsible-for-post-exposure-cellular-swelling-and-blebbing
#3
Gleb P Tolstykh, Gary L Thompson, Hope T Beier, Zachary A Steelman, Bennett L Ibey
Cell swelling and blebbing has been commonly observed following nanosecond pulsed electric field (nsPEF) exposure. The hypothesized origin of these effects is nanoporation of the plasma membrane (PM) followed by transmembrane diffusion of extracellular fluid and disassembly of cortical actin structures. This investigation will provide evidence that shows passive movement of fluid into the cell through nanopores and increase of intracellular osmotic pressure are not solely responsible for this observed phenomena...
March 2017: Biochemistry and Biophysics Reports
https://www.readbyqxmd.com/read/28917183/visualisation-of-an-nspef-induced-calcium-wave-using-the-genetically-encoded-calcium-indicator-gcamp-in-u87-human-glioblastoma-cells
#4
Lynn Carr, Sylvia M Bardet, Delia Arnaud-Cormos, Philippe Leveque, Rodney P O'Connor
Cytosolic, synthetic chemical calcium indicators are typically used to visualise the rapid increase in intracellular calcium ion concentration that follows nanosecond pulsed electric field (nsPEF) application. This study looks at the application of genetically encoded calcium indicators (GECIs) to investigate the spatiotemporal nature of nsPEF-induced calcium signals using fluorescent live cell imaging. Calcium responses to 44kV/cm, 10ns pulses were observed in U87-MG cells expressing either a plasma membrane targeted GECI (GCaMP5-G), or one cytosolically expressed (GCaMP6-S), and compared to the response of cells loaded with cytosolic or plasma membrane targeted chemical calcium indicators...
September 8, 2017: Bioelectrochemistry
https://www.readbyqxmd.com/read/28887559/delayed-hypersensitivity-to-nanosecond-pulsed-electric-field-in-electroporated-cells
#5
Sarah D Jensen, Vera A Khorokhorina, Claudia Muratori, Andrei G Pakhomov, Olga N Pakhomova
We demonstrate that conditioning of mammalian cells by electroporation with nanosecond pulsed electric field (nsPEF) facilitates their response to the next nsPEF treatment. The experiments were designed to unambiguously separate the electroporation-induced sensitization and desensitization effects. Electroporation was achieved by bursts of 300-ns, 9 kV/cm pulses (50 Hz, n = 3-100) and quantified by propidium dye uptake within 11 min after the nsPEF exposure. We observed either sensitization to nsPEF or no change (when the conditioning was either too weak or too intense, or when the wait time after conditioning was too short)...
September 8, 2017: Scientific Reports
https://www.readbyqxmd.com/read/28874684/damage-free-peripheral-nerve-stimulation-by-12-ns-pulsed-electric-field
#6
Maura Casciola, Shu Xiao, Andrei G Pakhomov
Modern technologies enable deep tissue focusing of nanosecond pulsed electric field (nsPEF) for non-invasive nerve and muscle stimulation. However, it is not known if PEF orders of magnitude shorter than the activation time of voltage-gated sodium channels (VGSC) would evoke action potentials (APs). One plausible scenario requires the loss of membrane integrity (electroporation) and resulting depolarization as an intermediate step. We report, for the first time, that the excitation of a peripheral nerve can be accomplished by 12-ns PEF without electroporation...
September 5, 2017: Scientific Reports
https://www.readbyqxmd.com/read/28693898/nanosecond-pulsed-electric-fields-depolarize-transmembrane-potential-via-voltage-gated-k-ca-2-and-trpm8-channels-in-u87-glioblastoma-cells
#7
Ryan C Burke, Sylvia M Bardet, Lynn Carr, Sergii Romanenko, Delia Arnaud-Cormos, Philippe Leveque, Rodney P O'Connor
Nanosecond pulsed electric fields (nsPEFs) have a variety of applications in the biomedical and biotechnology industries. Cancer treatment has been at the forefront of investigations thus far as nsPEFs permeabilize cellular and intracellular membranes leading to apoptosis and necrosis. nsPEFs may also influence ion channel gating and have the potential to modulate cell physiology without poration of the membrane. This phenomenon was explored using live cell imaging and a sensitive fluorescent probe of transmembrane voltage in the human glioblastoma cell line, U87 MG, known to express a number of voltage-gated ion channels...
October 2017: Biochimica et Biophysica Acta
https://www.readbyqxmd.com/read/28680807/calcium-dependent-activation-of-transglutaminase-2-by-nanosecond-pulsed-electric-fields
#8
Keiko Morotomi-Yano, Ken-Ichi Yano
Exposure of cultured human cells to nanosecond pulsed electric fields (nsPEFs) elicits various cellular events, including Ca(2+) influx and cell death. Recently, nsPEFs have been regarded as a novel physical treatment useful for biology and medicine, but the underlying mechanism of action remains to be fully elucidated. In this study, we investigated the effect of nsPEFs on transglutaminases (TGs), enzymes that catalyze covalent protein modifications such as protein-protein crosslinking. Cellular TG activity was monitored by conjugation of cellular proteins with biotin-cadaverine, a cell-permeable pseudosubstrate for TGs...
July 2017: FEBS Open Bio
https://www.readbyqxmd.com/read/28674780/delivery-devices-for-exposure-of-biological-cells-to-nanosecond-pulsed-electric-fields
#9
Malak Soueid, Martinus C F Dobbelaar, Sabrina Bentouati, Sylvia M Bardet, Rodney P O'Connor, Delphine Bessières, Jean Paillol, Philippe Leveque, Delia Arnaud-Cormos
In this paper, delivery devices for nanosecond pulsed electric field exposure of biological samples in direct contact with electrodes or isolated are presented and characterized. They are based on a modified electroporation cuvette and two transverse electromagnetic cells (TEM cells). The devices were used to apply pulses with high intensity (4.5 kV) and short durations (3 and 13 ns). The delivery devices were electromagnetically characterized in the frequency and time domains. Field intensities of around 5, 0...
July 4, 2017: Medical & Biological Engineering & Computing
https://www.readbyqxmd.com/read/28674451/novel-interventional-management-of-hepatic-hydatid-cyst-with-nanosecond-pulses-on-experimental-mouse-model
#10
Xinhua Chen, Ruiqing Zhang, Tuerganaili Aji, Yingmei Shao, Yonggang Chen, Hao Wen
The nanosecond pulsed electric field (nsPEF) is investigated as an alternative plan for benign hepatic hydatid cyst. Altogether 72 C57B6 mice were included. Normal group (n = 12) had no parasite injection and the other 60 mice were used to induce hydatid cyst in liver by injecting protoscolices in portal vein. The liver hydatid cysts were exposed to nsPEF with different doses and then follow up. The standard surgery was performed as positive control. The hydatid cyst growth was monitored by ultrasound; the morphology was checked by gross anatomy and pathology was tested by H&E stain...
July 3, 2017: Scientific Reports
https://www.readbyqxmd.com/read/28494437/nanosecond-pulsed-electric-field-induced-changes-in-cell-surface-charge-density
#11
Diganta Dutta, Xavier-Lewis Palmer, Anthony Asmar, Michael Stacey, Shizhi Qian
This study reports that the surface charge density changes in Jurkat cells with the application of single 60 nanosecond pulse electric fields, using atomic force microscopy. Using an atomic force microscope tip and Jurkat cells on silica in a 0.01M KCl ionic concentration, we were able to measure the interfacial forces, while also predicting surface charge densities of both Jurkat cell and silica surfaces. The most important finding is that the pulsing conditions varyingly reduced the cells' surface charge density...
September 2017: Micron: the International Research and Review Journal for Microscopy
https://www.readbyqxmd.com/read/28476039/nano-pulse-stimulation-nps-ablate-tumors-and-inhibit-lung-metastasis-on-both-canine-spontaneous-osteosarcoma-and-murine-transplanted-hepatocellular-carcinoma-with-high-metastatic-potential
#12
Xinhua Chen, Yiling Chen, Jianwen Jiang, Liming Wu, Shengyong Yin, Xudong Miao, Robert J Swanson, Shusen Zheng
BACKGROUND: Nanosecond pulsed electric field (nsPEF), which is also termed as nano-pulse stimulation (NPS), has the potential of stimulating immune responses toward cancer cells. The current study investigates its local and systemic antitumor efficacy in vivo in late stage tumors with lung metastasis. METHOD: The 12 canines with spontaneous osteosarcomas and 12 nude mice transplanted with human hepatocellular carcinoma were divided randomly and were given NPS treatment, surgery or no treatment control...
July 4, 2017: Oncotarget
https://www.readbyqxmd.com/read/28467607/environmental-temperature-affects-physiology-and-survival-of-nanosecond-pulsed-electric-field-treated-cells
#13
COMPARATIVE STUDY
Shengyong Yin, Xudong Miao, Xueming Zhang, Xinhua Chen, Hao Wen
Nanosecond pulsed electric field (nsPEF) is a novel non-thermal tumor ablation technique. However, how nsPEF affect cell physiology at different environmental temperature is still kept unknown. But this issue is of critical clinical practice relevance. This work aim to investigate how nsPEF treated cancer cells react to different environmental temperatures (0, 4, 25, and 37°C). Their cell viability, apoptosis, mitochondrial membrane potential, and reactive oxygen species (ROS) were examined. Lower temperature resulted in higher apoptosis rate, decreased mitochondria membrane potential, and increased ROS levels...
February 2018: Journal of Cellular Physiology
https://www.readbyqxmd.com/read/28432032/neuronal-excitation-and-permeabilization-by-200-ns-pulsed-electric-field-an-optical-membrane-potential-study-with-fluovolt-dye
#14
Andrei G Pakhomov, Iurii Semenov, Maura 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...
July 2017: Biochimica et Biophysica Acta
https://www.readbyqxmd.com/read/28393078/technological-and-theoretical-aspects-for-testing-electroporation-on-liposomes
#15
Agnese Denzi, Elena Della Valle, Gianluca Esposito, Lluis M Mir, Francesca Apollonio, Micaela Liberti
Recently, the use of nanometer liposomes as nanocarriers in drug delivery systems mediated by nanoelectroporation has been proposed. This technique takes advantage of the possibility of simultaneously electroporating liposomes and cell membrane with 10-nanosecond pulsed electric fields (nsPEF) facilitating the release of the drug from the liposomes and at the same time its uptake by the cells. In this paper the design and characterization of a 10 nsPEF exposure system is presented, for liposomes electroporation purposes...
2017: BioMed Research International
https://www.readbyqxmd.com/read/28365791/elasticity-and-tumorigenic-characteristics-of-cells-in-a-monolayer-after-nanosecond-pulsed-electric-field-exposure
#16
A Steuer, K Wende, P Babica, J F Kolb
Nanosecond pulsed electric fields (nsPEFs) applied to cells can induce different biological effects depending on pulse duration and field strength. One known process is the induction of apoptosis whereby nsPEFs are currently investigated as a novel cancer therapy. Another and probably related change is the breakdown of the cytoskeleton. We investigated the elasticity of rat liver epithelial cells WB-F344 in a monolayer using atomic force microscopy (AFM) with respect to the potential of cells to undergo malignant transformation or to develop a potential to metastasize...
September 2017: European Biophysics Journal: EBJ
https://www.readbyqxmd.com/read/28236173/nanosecond-pulsed-electric-field-nspef-disrupts-the-structure-and-metabolism-of-human-echinococcus-granulosus-protoscolex-in-vitro-with-a-dose-effect
#17
Ruiqing Zhang, Tuerganaili Aji, Yingmei Shao, Tiemin Jiang, Lei Yang, Weimin Lv, Yonggang Chen, Xinhua Chen, Hao Wen
The number of interventional treatments for hepatic cystic echinococcosis is increasing, but the chemicals or high temperatures used in these methodologies cause biliary complications, thus limiting their clinical applications. This experimental study aimed to apply a novel, non-thermal, non-chemical ablation method termed nanosecond pulsed electric field (nsPEF) for the treatment of human hepatic cystic echinococcosis. The nsPEF treatment parameters against protoscolices from human hepatic cystic echinococcosis were optimized in vitro...
April 2017: Parasitology Research
https://www.readbyqxmd.com/read/28228843/a-new-mechanism-for-efficient-hydrocarbon-electro-extraction-from-botryococcus-braunii
#18
Alexis Guionet, Bahareh Hosseini, Justin Teissié, Hidenori Akiyama, Hamid Hosseini
BACKGROUND: Recent understanding that specific algae have high hydrocarbon production potential has attracted considerable attention. Botryococcus braunii is a microalga with an extracellular hydrocarbon matrix, which makes it an appropriate green energy source. RESULTS: This study focuses on extracting oil from the microalgae matrix rather than the cells, eliminating the need for an excessive electric field to create electro-permeabilization. In such a way, technical limitations due to high extraction energy and cost can be overcome...
2017: Biotechnology for Biofuels
https://www.readbyqxmd.com/read/28147680/miniaturized-two-stack-blumlein-pulser-with-a-variable-repetition-rate-for-non-thermal-irreversible-electroporation-experiments
#19
Sun-Hong Min, Ohjoon Kwon, Matlabjon Sattorov, In-Keun Baek, Seontae Kim, Jin-Young Jeong, Dongpyo Hong, Seunghyuk Park, Gun-Sik Park
Non-thermal irreversible electroporation (NTIRE) to avoid thermal damage to cells during intense DC ns pulsed electric fields (nsPEFs) is a recent modality for medical applications. This mechanism, related to bioelectrical dynamics of the cell, is linked to the effect of a DC electric field and a threshold effect with an electrically stimulated membrane for the charge distribution in the cell. To create the NTIRE condition, the pulse width of the nsPEF should be shorter than the charging time constant of the membrane related to the cell radius, membrane capacitance, cytoplasm resistivity, and medium resistivity...
January 2017: Review of Scientific Instruments
https://www.readbyqxmd.com/read/28117459/calcium-independent-disruption-of-microtubule-dynamics-by-nanosecond-pulsed-electric-fields-in-u87-human-glioblastoma-cells
#20
Lynn Carr, Sylvia M Bardet, Ryan C Burke, Delia Arnaud-Cormos, Philippe Leveque, Rodney P O'Connor
High powered, nanosecond duration, pulsed electric fields (nsPEF) cause cell death by a mechanism that is not fully understood and have been proposed as a targeted cancer therapy. Numerous chemotherapeutics work by disrupting microtubules. As microtubules are affected by electrical fields, this study looks at the possibility of disrupting them electrically with nsPEF. Human glioblastoma cells (U87-MG) treated with 100, 10 ns, 44 kV/cm pulses at a frequency of 10 Hz showed a breakdown of their interphase microtubule network that was accompanied by a reduction in the number of growing microtubules...
January 24, 2017: Scientific Reports
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