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Sample records for selective endosomal ablation

  1. Selective Laser Ablation and Melting, Phase I

    Data.gov (United States)

    National Aeronautics and Space Administration — In this project Advratech will develop a new additive manufacturing (AM) process called Selective Laser Ablation and Melting (SLAM). The key innovation in this...

  2. Selective corneal optical aberration (SCOA) for customized ablation

    Science.gov (United States)

    Jean, Benedikt J.; Bende, Thomas

    2001-06-01

    Wavefront analysis still have some technical problems which may be solved within the next years. There are some limitations to use wavefront as a diagnostic tool for customized ablation alone. An ideal combination would be wavefront and topography. Meanwhile Selective Corneal Aberration is a method to visualize the optical quality of a measured corneal surface. It is based on a true measured 3D elevation information of a video topometer. Thus values can be interpreted either using Zernike polynomials or visualized as a so called color coded surface quality map. This map gives a quality factor (corneal aberration) for each measured point of the cornea.

  3. Retromer guides STxB and CD8-M6PR from early to recycling endosomes, EHD1 guides STxB from recycling endosome to Golgi

    Science.gov (United States)

    McKenzie, Jenna E.; Raisley, Brent; Zhou, Xin; Naslavsky, Naava; Taguchi, Tomohiko; Caplan, Steve; Sheff, David

    2012-01-01

    Retrograde trafficking transports proteins, lipids and toxins from the plasma membrane to the Golgi and ER. To reach the Golgi, these cargos must transit the endosomal system, consisting of early endosomes, recycling endosomes, late endosomes and lysosomes. All cargos pass through early endosomes, but may take different routes to the Golgi. Retromer dependent cargos bypass the late endosomes to reach the Golgi. We compared how two very different retromer dependent cargos negotiate the endosomal sorting system. Shiga toxin B, bound to the external layer of the plasma membrane, and chimeric CD8-Mannose-6-Phosphate Receptor, which is anchored via a transmembrane domain. Both appear to pass through the recycling endosome. Ablation of the recycling endosome diverted both of these cargos to an aberrant compartment and prevented them from reaching the Golgi. Once in the recycling endosome, Shiga toxin required EHD1 to traffic to the TGN, while the CD8-Mannose-6-Phosphate Receptor was not significantly dependent on EHD1. Knockdown of retromer components left cargo in the early endosomes, suggesting that it is required for retrograde exit from this compartment. This work establishes the recycling endosome as a required step in retrograde traffic of at least these two retromer dependent cargos. Along this pathway, retromer is associated with EE to recycling endosome traffic, while EHD1 is associated with recycling endosome to TGN traffic of STxB. PMID:22540229

  4. Alantolactone selectively ablates acute myeloid leukemia stem and progenitor cells

    Directory of Open Access Journals (Sweden)

    Yahui Ding

    2016-09-01

    Full Text Available Abstract Background The poor outcomes for patients diagnosed with acute myeloid leukemia (AML are largely attributed to leukemia stem cells (LSCs which are difficult to eliminate with conventional therapy and responsible for relapse. Thus, new therapeutic strategies which could selectively target LSCs in clinical leukemia treatment and avoid drug resistance are urgently needed. However, only a few small molecules have been reported to show anti-LSCs activity. Methods The aim of the present study was to identify alantolactone as novel agent that can ablate acute myeloid leukemia stem and progenitor cells from AML patient specimens and evaluate the anticancer activity of alantolactone in vitro and in vivo. Results The present study is the first to demonstrate that alantolactone, a prominent eudesmane-type sesquiterpene lactone, could specifically ablate LSCs from AML patient specimens. Furthermore, in comparison to the conventional chemotherapy drug, cytosine arabinoside (Ara-C, alantolactone showed superior effects of leukemia cytotoxicity while sparing normal hematopoietic cells. Alantolactone induced apoptosis with a dose-dependent manner by suppression of NF-kB and its downstream target proteins. DMA-alantolactone, a water-soluble prodrug of alantolactone, could suppress tumor growth in vivo. Conclusions Based on these results, we propose that alantolactone may represent a novel LSCs-targeted therapy and eudesmane-type sesquiterpene lactones offer a new scaffold for drug discovery towards anti-LSCs agents.

  5. Ablation of selected conducting layers by fiber laser

    Science.gov (United States)

    Pawlak, Ryszard; Tomczyk, Mariusz; Walczak, Maria

    2014-08-01

    Laser Direct Writing (LDW) are used in the manufacture of electronic circuits, pads, and paths in sub millimeter scale. They can also be used in the sensors systems. Ablative laser writing in a thin functional layer of material deposited on the dielectric substrate is one of the LDW methods. Nowadays functional conductive layers are composed from graphene paint or nanosilver paint, indium tin oxide (ITO), AgHTTM and layers containing carbon nanotubes. Creating conducting structures in transparent layers (ITO, AgHT and carbon nanotubes layers) may have special importance e.g. for flexi electronics. The paper presents research on the fabrication of systems of paths and appropriate pattern systems of paths and selected electronic circuits in AgHTTM and ITO layers deposited on glass and polymer substrates. An influence of parameters of ablative fiber laser treatment in nanosecond regime as well as an influence of scanning mode of laser beam on the pattern fidelity and on electrical parameters of a generated circuit was investigated.

  6. Alcohol septal ablation: patient selection and rationality of its application

    Directory of Open Access Journals (Sweden)

    М. Г. Каштанов

    2017-04-01

    Full Text Available The article looks at some aspects of selecting patients with obstructive hypertrophic cardiomyopathy for alcohol septal ablation (ASA procedure. Based on the world’s experience and in the context of evidence-based medicine, the current positions of ASA in complex treatment of obstructive hypertrophic cardiomyopathy are described. Received 29 September 2016. Accepted 9 January 2017.Funding: The study had no sponsorship.Conflict of interest: The authors declare no conflict of interest.Author contributionsData collection and analysis: Kashtanov M.G. Drafting the article: Kashtanov M.G. Critical revision: Kashtanov M.G., Idov E.M., Chernyshev S.D., Kardapoltsev L.V., Berdnikov S.V.

  7. High resolution selective multilayer laser processing by nanosecond laser ablation of metal nanoparticle films

    International Nuclear Information System (INIS)

    Ko, Seung H.; Pan Heng; Hwang, David J.; Chung, Jaewon; Ryu, Sangil; Grigoropoulos, Costas P.; Poulikakos, Dimos

    2007-01-01

    Ablation of gold nanoparticle films on polymer was explored using a nanosecond pulsed laser, with the goal to achieve feature size reduction and functionality not amenable with inkjet printing. The ablation threshold fluence for the unsintered nanoparticle deposit was at least ten times lower than the reported threshold for the bulk film. This could be explained by the combined effects of melting temperature depression, lower conductive heat transfer loss, strong absorption of the incident laser beam, and the relatively weak bonding between nanoparticles. The ablation physics were verified by the nanoparticle sintering characterization, ablation threshold measurement, time resolved ablation plume shadowgraphs, analysis of ablation ejecta, and the measurement and calculation of optical properties. High resolution and clean feature fabrication with small energy and selective multilayer processing are demonstrated

  8. Stress assisted selective ablation of ITO thin film by picosecond laser

    Science.gov (United States)

    Farid, Nazar; Chan, Helios; Milne, David; Brunton, Adam; M. O'Connor, Gerard

    2018-01-01

    Fast selective pattering with high precession on 175 nm ITO thin film with IR ps lasers is investigated. Ablation parameters are optimized with detailed studies on the scribed depth, topography, and particle generation using AFM and SEM. A comparison of 10 and 150 ps laser revealed that the shorter pulse (10 ps) laser is more appropriate in selective and partial ablation; up to 20 nm resolution for controlled depth with multipulses having energy below the damage threshold is demonstrated. The experimental results are interpreted to involve stress assisted ablation mechanism for the 10 ps laser while thermal ablation along with intense melting occurs for 150 ps laser. The transition between these regimes is estimated to occur at approximately 30 ps.

  9. Considering Angle Selection When Using Ultrasound Electrode Displacement Elastography to Evaluate Radiofrequency Ablation of Tissues

    Science.gov (United States)

    Li, Qiang; Chen, Pin-Yu; Wang, Chiao-Yin; Liu, Hao-Li; Teng, Jianfu

    2014-01-01

    Percutaneous radiofrequency ablation (RFA) is a minimally invasive treatment to thermally destroy tumors. Ultrasound-based electrode-displacement elastography is an emerging technique for evaluating the region of RFA-induced lesions. The angle between the imaging probe and the RFA electrode can influence electrode-displacement elastography when visualizing the ablation zone. We explored the angle effect on electrode-displacement elastography to measure the ablation zone. Phantoms embedded with meatballs were fabricated and then ablated using an RFA system to simulate RFA-induced lesions. For each phantom, a commercial ultrasound scanner with a 7.5 MHz linear probe was used to acquire raw image data at different angles, ranging from 30° to 90° at increments of 10°, to construct electrode-displacement images and facilitate comparisons with tissue section images. The results revealed that the ablation regions detected using electrode-displacement elastography were highly correlated with those from tissue section images when the angle was between 30° and 60°. However, the boundaries of lesions were difficult to distinguish, when the angle was larger than 60°. The experimental findings suggest that angle selection should be considered to achieve reliable electrode-displacement elastography to describe ablation zones. PMID:24971347

  10. Optical fiber biocompatible sensors for monitoring selective treatment of tumors via thermal ablation

    Science.gov (United States)

    Tosi, Daniele; Poeggel, Sven; Dinesh, Duraibabu B.; Macchi, Edoardo G.; Gallati, Mario; Braschi, Giovanni; Leen, Gabriel; Lewis, Elfed

    2015-09-01

    Thermal ablation (TA) is an interventional procedure for selective treatment of tumors, that results in low-invasive outpatient care. The lack of real-time control of TA is one of its main weaknesses. Miniature and biocompatible optical fiber sensors are applied to achieve a dense, multi-parameter monitoring, that can substantially improve the control of TA. Ex vivo measurements are reported performed on porcine liver tissue, to reproduce radiofrequency ablation of hepatocellular carcinoma. Our measurement campaign has a two-fold focus: (1) dual pressure-temperature measurement with a single probe; (2) distributed thermal measurement to estimate point-by-point cells mortality.

  11. Selective laser etching or ablation for fabrication of devices

    KAUST Repository

    Buttner, Ulrich; Salama, Khaled N.; Sapsanis, Christos

    2017-01-01

    Methods of fabricating devices vial selective laser etching are provided. The methods can include selective laser etching of a portion of a metal layer, e.g. using a laser light source having a wavelength of 1,000 nm to 1,500 nm. The methods can

  12. Selective laser etching or ablation for fabrication of devices

    KAUST Repository

    Buttner, Ulrich

    2017-01-12

    Methods of fabricating devices vial selective laser etching are provided. The methods can include selective laser etching of a portion of a metal layer, e.g. using a laser light source having a wavelength of 1,000 nm to 1,500 nm. The methods can be used to fabricate a variety of features, including an electrode, an interconnect, a channel, a reservoir, a contact hole, a trench, a pad, or a combination thereof. A variety of devices fabricated according to the methods are also provided. In some aspects, capacitive humidity sensors are provided that can be fabricated according to the provided methods. The capacitive humidity sensors can be fabricated with intricate electrodes, e.g. having a fractal pattern such as a Peano curve, a Hilbert curve, a Moore curve, or a combination thereof.

  13. Selective removal of esthetic composite restorations with spectral guided laser ablation

    Science.gov (United States)

    Yi, Ivana; Chan, Kenneth H.; Tsuji, Grant H.; Staninec, Michal; Darling, Cynthia L.; Fried, Daniel

    2016-02-01

    Dental composites are used for a wide range of applications such as fillings for cavities, adhesives for orthodontic brackets, and closure of gaps (diastemas) between teeth by esthetic bonding. Anterior restorations are used to replace missing, diseased and unsightly tooth structure for both appearance and function. When these restorations must be replaced, they are difficult to remove mechanically without causing excessive removal or damage to enamel because dental composites are color matched to teeth. Previous studies have shown that CO2 lasers have high ablation selectivity and are well suited for removal of composite on occlusal surfaces while minimizing healthy tissue loss. A spectral feedback guidance system may be used to discriminate between dental composite and dental hard tissue for selective ablation of composite material. The removal of composite restorations filling diastemas is more challenging due to the esthetic concern for anterior teeth. The objective of this study is to determine if composite spanning a diastema between anterior teeth can be removed by spectral guided laser ablation at clinically relevant rates with minimal damage to peripheral healthy tissue and with higher selectivity than a high speed dental handpiece.

  14. The efficacy of selective calculus ablation at 400 nm: comparison to conventional calculus removal methods

    Science.gov (United States)

    Schoenly, Joshua E.; Seka, Wolf; Romanos, Georgios; Rechmann, Peter

    A desired outcome of scaling and root planing is the complete removal of calculus and infected root tissue and preservation of healthy cementum for rapid healing of periodontal tissues. Conventional periodontal treatments for calculus removal, such as hand instrument scaling and ultrasonic scaling, often deeply scrape the surface of the underlying hard tissue and may leave behind a smear layer. Pulsed lasers emitting at violet wavelengths (specifically, 380 to 400 nm) are a potential alternative treatment since they can selectively ablate dental calculus without ablating pristine hard tissue (i.e., enamel, cementum, and dentin). In this study, light and scanning electron microscopy are used to compare and contrast the efficacy of in vitro calculus removal for several conventional periodontal treatments (hand instruments, ultrasonic scaler, and Er:YAG laser) to calculus removal with a frequency-doubled Ti:sapphire (λ = 400 nm). After calculus removal, enamel and cementum surfaces are investigated for calculus debris and damage to the underlying hard tissue surface. Compared to the smear layer, grooves, and unintentional hard tissue removal typically found using these conventional treatments, calculus removal using the 400-nm laser is complete and selective without any removal of pristine dental hard tissue. Based on these results, selective ablation from the 400-nm laser appears to produce a root surface that would be more suitable for successful healing of periodontal tissues.

  15. Dynamic frame selection for in vivo ultrasound temperature estimation during radiofrequency ablation

    International Nuclear Information System (INIS)

    Daniels, Matthew J; Varghese, Tomy

    2010-01-01

    Minimally invasive therapies such as radiofrequency ablation have been developed to treat cancers of the liver, prostate and kidney without invasive surgery. Prior work has demonstrated that ultrasound echo shifts due to temperature changes can be utilized to track the temperature distribution in real time. In this paper, a motion compensation algorithm is evaluated to reduce the impact of cardiac and respiratory motion on ultrasound-based temperature tracking methods. The algorithm dynamically selects the next suitable frame given a start frame (selected during the exhale or expiration phase where extraneous motion is reduced), enabling optimization of the computational time in addition to reducing displacement noise artifacts incurred with the estimation of smaller frame-to-frame displacements at the full frame rate. A region of interest that does not undergo ablation is selected in the first frame and the algorithm searches through subsequent frames to find a similarly located region of interest in subsequent frames, with a high value of the mean normalized cross-correlation coefficient value. In conjunction with dynamic frame selection, two different two-dimensional displacement estimation algorithms namely a block matching and multilevel cross-correlation are compared. The multi-level cross-correlation method incorporates tracking of the lateral tissue expansion in addition to the axial deformation to improve the estimation performance. Our results demonstrate the ability of the proposed motion compensation using dynamic frame selection in conjunction with the two-dimensional multilevel cross-correlation to track the temperature distribution.

  16. Selective ablation of photovoltaic materials with UV laser sources for monolithic interconnection of devices based on a-Si:H

    Energy Technology Data Exchange (ETDEWEB)

    Molpeceres, C. [Centro Laser UPM, Univ. Politecnica de Madrid, Crta. de Valencia Km 7.3, 28031 Madrid (Spain)], E-mail: carlos.molpeceres@upm.es; Lauzurica, S.; Garcia-Ballesteros, J.J.; Morales, M.; Guadano, G.; Ocana, J.L. [Centro Laser UPM, Univ. Politecnica de Madrid, Crta. de Valencia Km 7.3, 28031 Madrid (Spain); Fernandez, S.; Gandia, J.J. [Dept. de Energias Renovables, Energia Solar Fotovoltaica, CIEMAT, Avda, Complutense 22, 28040 Madrid (Spain); Villar, F.; Nos, O.; Bertomeu, J. [CeRMAE Dept. Fisica Aplicada i Optica, Universitat de Barcelona, Av. Diagonal 647, 08028 Barcelona (Spain)

    2009-03-15

    Lasers are essential tools for cell isolation and monolithic interconnection in thin-film-silicon photovoltaic technologies. Laser ablation of transparent conductive oxides (TCOs), amorphous silicon structures and back contact removal are standard processes in industry for monolithic device interconnection. However, material ablation with minimum debris and small heat affected zone is one of the main difficulty is to achieve, to reduce costs and to improve device efficiency. In this paper we present recent results in laser ablation of photovoltaic materials using excimer and UV wavelengths of diode-pumped solid-state (DPSS) laser sources. We discuss results concerning UV ablation of different TCO and thin-film silicon (a-Si:H and nc-Si:H), focussing our study on ablation threshold measurements and process-quality assessment using advanced optical microscopy techniques. In that way we show the advantages of using UV wavelengths for minimizing the characteristic material thermal affection of laser irradiation in the ns regime at higher wavelengths. Additionally we include preliminary results of selective ablation of film on film structures irradiating from the film side (direct writing configuration) including the problem of selective ablation of ZnO films on a-Si:H layers. In that way we demonstrate the potential use of UV wavelengths of fully commercial laser sources as an alternative to standard backscribing process in device fabrication.

  17. Selective ablation of photovoltaic materials with UV laser sources for monolithic interconnection of devices based on a-Si:H

    International Nuclear Information System (INIS)

    Molpeceres, C.; Lauzurica, S.; Garcia-Ballesteros, J.J.; Morales, M.; Guadano, G.; Ocana, J.L.; Fernandez, S.; Gandia, J.J.; Villar, F.; Nos, O.; Bertomeu, J.

    2009-01-01

    Lasers are essential tools for cell isolation and monolithic interconnection in thin-film-silicon photovoltaic technologies. Laser ablation of transparent conductive oxides (TCOs), amorphous silicon structures and back contact removal are standard processes in industry for monolithic device interconnection. However, material ablation with minimum debris and small heat affected zone is one of the main difficulty is to achieve, to reduce costs and to improve device efficiency. In this paper we present recent results in laser ablation of photovoltaic materials using excimer and UV wavelengths of diode-pumped solid-state (DPSS) laser sources. We discuss results concerning UV ablation of different TCO and thin-film silicon (a-Si:H and nc-Si:H), focussing our study on ablation threshold measurements and process-quality assessment using advanced optical microscopy techniques. In that way we show the advantages of using UV wavelengths for minimizing the characteristic material thermal affection of laser irradiation in the ns regime at higher wavelengths. Additionally we include preliminary results of selective ablation of film on film structures irradiating from the film side (direct writing configuration) including the problem of selective ablation of ZnO films on a-Si:H layers. In that way we demonstrate the potential use of UV wavelengths of fully commercial laser sources as an alternative to standard backscribing process in device fabrication.

  18. Selective ex-vivo photothermal ablation of human pancreatic cancer with albumin functionalized multiwalled carbon nanotubes.

    Science.gov (United States)

    Mocan, Lucian; Tabaran, Flaviu A; Mocan, Teodora; Bele, Constantin; Orza, Anamaria Ioana; Lucan, Ciprian; Stiufiuc, Rares; Manaila, Ioana; Iulia, Ferencz; Dana, Iancu; Zaharie, Florin; Osian, Gelu; Vlad, Liviu; Iancu, Cornel

    2011-01-01

    The process of laser-mediated ablation of cancer cells marked with biofunctionalized carbon nanotubes is frequently called "nanophotothermolysis". We herein present a method of selective nanophotothermolisys of pancreatic cancer (PC) using multiwalled carbon nanotubes (MWCNTs) functionalized with human serum albumin (HSA). With the purpose of testing the therapeutic value of these nanobioconjugates, we have developed an ex-vivo experimental platform. Surgically resected specimens from patients with PC were preserved in a cold medium and kept alive via intra-arterial perfusion. Additionally, the HSA-MWCNTs have been intra-arterially administered in the greater pancreatic artery under ultrasound guidance. Confocal and transmission electron microscopy combined with immunohistochemical staining have confirmed the selective accumulation of HSA-MWCNTs inside the human PC tissue. The external laser irradiation of the specimen has significantly produced extensive necrosis of the malign tissue after the intra-arterial administration of HSA-MWCNTs, without any harmful effects on the surrounding healthy parenchyma. We have obtained a selective photothermal ablation of the malign tissue based on the selective internalization of MWCNTs with HSA cargo inside the pancreatic adenocarcinoma after the ex-vivo intra-arterial perfusion.

  19. Internal structure of magnetic endosomes

    Science.gov (United States)

    Rivière, C.; Wilhelm, C.; Cousin, F.; Dupuis, V.; Gazeau, F.; Perzynski, R.

    2007-01-01

    The internal structure of biological vesicles filled with magnetic nanoparticles is investigated using the following complementary analyses: electronic transmission microscopy, dynamic probing by magneto-optical birefringence and structural probing by Small Angle Neutron Scattering (SANS). These magnetic vesicles are magnetic endosomes obtained via a non-specific interaction between cells and anionic magnetic iron oxide nanoparticles. Thanks to a magnetic purification process, they are probed at two different stages of their formation within HeLa cells: (i) adsorption of nanoparticles onto the cellular membrane and (ii) their subsequent internalisation within endosomes. Differences in the microenvironment of the magnetic nanoparticles at those two different stages are highlighted here. The dynamics of magnetic nanoparticles adsorbed onto cellular membranes and confined within endosomes is respectively 3 and 5 orders of magnitude slower than for isolated magnetic nanoparticles in aqueous media. Interestingly, SANS experiments show that magnetic endosomes have an internal structure close to decorated vesicles, with magnetic nanoparticles locally decorating the endosome membrane, inside their inner-sphere. These results, important for future biomedical applications, suggest that multiple fusions of decorated vesicles are the biological processes underlying the endocytosis of that kind of nanometric materials.

  20. Selective ex-vivo photothermal ablation of human pancreatic cancer with albumin functionalized multiwalled carbon nanotubes

    Directory of Open Access Journals (Sweden)

    Mocan L

    2011-04-01

    Full Text Available Lucian Mocan1, Flaviu A Tabaran2, Teodora Mocan1, Constantin Bele3, Anamaria Ioana Orza1, Ciprian Lucan4, Rares Stiufiuc1, Ioana Manaila1, Ferencz Iulia1, Iancu Dana1, Florin Zaharie1, Gelu Osian1, Liviu Vlad1, Cornel Iancu11Department of Nanomedicine, “Iuliu Hatieganu” University of Medicine and Pharmacy, Cluj-Napoca, Romania; 2Department of Pathology, University of Agricultural Sciences and Veterinary Medicine, Cluj-Napoca, Romania; 3Department of Biochemistry, University of Agricultural Sciences and Veterinary Medicine, Cluj-Napoca, Romania; 4Clinical Institute of Urology and Renal Transplantation, Cluj-Napoca, RomaniaAbstract: The process of laser-mediated ablation of cancer cells marked with biofunctionalized carbon nanotubes is frequently called “nanophotothermolysis”. We herein present a method of selective nanophotothermolisys of pancreatic cancer (PC using multiwalled carbon nanotubes (MWCNTs functionalized with human serum albumin (HSA. With the purpose of testing the therapeutic value of these nanobioconjugates, we have developed an ex-vivo experimental platform. Surgically resected specimens from patients with PC were preserved in a cold medium and kept alive via intra-arterial perfusion. Additionally, the HSA-MWCNTs have been intra-arterially administered in the greater pancreatic artery under ultrasound guidance. Confocal and transmission electron microscopy combined with immunohistochemical staining have confirmed the selective accumulation of HSA-MWCNTs inside the human PC tissue. The external laser irradiation of the specimen has significantly produced extensive necrosis of the malign tissue after the intra-arterial administration of HSA-MWCNTs, without any harmful effects on the surrounding healthy parenchyma. We have obtained a selective photothermal ablation of the malign tissue based on the selective internalization of MWCNTs with HSA cargo inside the pancreatic adenocarcinoma after the ex-vivo intra

  1. Selective ablation of a titanium nitride film on tungsten carbide substrate using ultrashort laser pulses

    International Nuclear Information System (INIS)

    Oliveira, Eduardo Spinelli

    2017-01-01

    Surface coatings are applied to many cutting tools in the metallurgical industry in order to improve cutting efficiency and extend its useful life. In this work, tests were performed to remove the coating of titanium aluminum nitride (TiAlN) on tungsten carbide (WC-Co) pellets, using an ultrashort laser pulses beam. After determination of the damage thresholds of the film and the substrate, were ablated on the surface of the coating lines using two ablation conditions, it was initially operated on the low fluence regime for the film, and later on the low fluence regime of the substrate, far below the threshold of the film, applying high overlapping pulses. A laser induced breakdown spectroscopy (LIBS) system was set up to monitor the materials present in the plasma generated by the laser, but the system did not present sufficient sensitivity to read the low intensity of the plasma generated in the process and was not used. After the analysis of the traces by electron microscopy, optical profilometer and X-ray fluorescence spectroscopy, it was not possible to determine a safe process to carry out the selective removal of the film in question, however, due to the data obtained and observations of the results in some traces, new possibilities were raised, opening the discussion for future work. (author)

  2. Membrane Tethering Complexes in the Endosomal System

    OpenAIRE

    Spang, Anne

    2016-01-01

    Vesicles that are generated by endocytic events at the plasma membrane are destined to early endosomes. A prerequisite for proper fusion is the tethering of two membrane entities. Tethering of vesicles to early endosomes is mediated by the class C core vacuole/endosome tethering (CORVET) complex, while fusion of late endosomes with lysosomes depends on the homotypic fusion and vacuole protein sorting (HOPS) complex. Recycling through the trans-Golgi network (TGN) and to the plasma membrane is...

  3. Endocytosis and Endosomal Trafficking in Plants.

    Science.gov (United States)

    Paez Valencia, Julio; Goodman, Kaija; Otegui, Marisa S

    2016-04-29

    Endocytosis and endosomal trafficking are essential processes in cells that control the dynamics and turnover of plasma membrane proteins, such as receptors, transporters, and cell wall biosynthetic enzymes. Plasma membrane proteins (cargo) are internalized by endocytosis through clathrin-dependent or clathrin-independent mechanism and delivered to early endosomes. From the endosomes, cargo proteins are recycled back to the plasma membrane via different pathways, which rely on small GTPases and the retromer complex. Proteins that are targeted for degradation through ubiquitination are sorted into endosomal vesicles by the ESCRT (endosomal sorting complex required for transport) machinery for degradation in the vacuole. Endocytic and endosomal trafficking regulates many cellular, developmental, and physiological processes, including cellular polarization, hormone transport, metal ion homeostasis, cytokinesis, pathogen responses, and development. In this review, we discuss the mechanisms that mediate the recognition and sorting of endocytic and endosomal cargos, the vesiculation processes that mediate their trafficking, and their connection to cellular and physiological responses in plants.

  4. Membrane tethering complexes in the endosomal system

    Directory of Open Access Journals (Sweden)

    Anne eSpang

    2016-05-01

    Full Text Available Vesicles that are generated by endocytic events at the plasma membrane are destined to early endosomes. A prerequisite for proper fusion is the tethering of two membrane entities. Tethering of vesicles to early endosomes is mediated by the CORVET complex, while fusion of late endosomes with lysosomes depends on the HOPS complex. Recycling through the TGN and to the plasma membrane is facilitated by the GARP and EARP complexes, respectively. However, there are other tethering functions in the endosomal system as there are multiple pathways through which proteins can be delivered from endosomes to either the TGN or the plasma membrane. Furthermore, complexes that may be part of novel tethering complexes have been recently identified. Thus it is likely that more tethering factors exist. In this review, I will provide an overview of different tethering complexes of the endosomal system and discuss how they may provide specificity in membrane traffic.

  5. Selective near-UV ablation of subgingival dental calculus: measurement of removal rates

    Science.gov (United States)

    Schoenly, J. E.; Seka, W.; Rechmann, P.

    2010-02-01

    A noncontact profilometer (laser triangulation) was used to measure the removal rates of subgingival dental calculus irradiated with a frequency-doubled Ti:sapphire laser (60-ns pulse duration, 400-nm wavelength, 10-Hz repetition rate, 7-mJ pulse energy). Profilometer traces before and after irradiation were used to create a removal map with 4-μm axial and 15-μm transverse resolution. Twenty-three teeth (15 with calculus and 8 pristine) were irradiated at 90° and 45° under a cooling water spray with a super-Gaussian beam (~300-μm diameter). Subgingival calculus was selectively removed at 5.6 and 4.0 J/cm2 for 90° and 45°, respecetively, within a range of rates, between 2 to 9 μm/pulse. These ablation rates were constant during these exposures. For comparison, pristine cementum irradiated for 10 min at the same peak fluence and pulse repetition rate showed only craters, 15 to 50 μm deep, corresponding to an equivalent removal rate three orders of magnitude smaller than that obtained for calculus. Pristine enamel was not removed under the same irradiation conditions.

  6. Useful clinical features for the selection of ideal patients with strial fibrillation for mapping and catheter ablation

    Directory of Open Access Journals (Sweden)

    Mehta Niraj

    2002-01-01

    Full Text Available OBJECTIVE: To identify useful clinical characteristics for selecting patients eligible for mapping and ablation of atrial fibrillation. METHODS: We studied 9 patients with atrial fibrillation, without structural heart disease, associated with: 1 antiarrhythmic drugs, 2 symptoms of low cardiac output, and 3 intention to treat. Seven patients had paroxysmal atrial fibrillation and 2 had recurrent atrial fibrillation. RESULTS: In the 6 patients who underwent mapping (all had paroxysmal atrial fibrillation, catheter ablation was successfully carried out in superior pulmonary veins in 5 patients (the first 3 in the left superior pulmonary vein and the last 2 in the right superior pulmonary vein. One patient experienced a recurrence of atrial fibrillation after 10 days. We observed that patients who had short episodes of atrial fibrillation on 24-hour Holter monitoring before the procedure were those in whom mapping the focus of tachycardia was possible. Tachycardia was successfully suppressed in 4 of 6 patients. The cause of failure was due to the impossibility of maintaining sinus rhythm long enough for efficient mapping. CONCLUSION: Patients experiencing short episodes of atrial fibrillation during 24-hour Holter monitoring were the most eligible for mapping and ablation, with a final success rate of 66%, versus the global success rate of 44%. Patients with persistent atrial fibrillation were not good candidates for focal ablation.

  7. Investigation into the optimum beam shape and fluence for selective ablation of dental calculus at lambda = 400 nm.

    Science.gov (United States)

    Schoenly, Joshua E; Seka, Wolf; Rechmann, Peter

    2010-01-01

    A frequency-doubled Ti:sapphire laser is shown to selectively ablate dental calculus. The optimal transverse shape of the laser beam, including its variability under water-cooling, is determined for selective ablation of dental calculus. Intensity profiles under various water-cooling conditions were optically observed. The 400-nm laser was coupled into a multimode optical fiber using an f = 2.5-cm lens and light-shaping diffuser. Water-cooling was supplied coaxially around the fiber. Five human tooth samples (four with calculus and one pristine) were irradiated perpendicular to the tooth surface while the tooth was moved back and forth at 0.3 mm/second, varying between 20 and 180 iterations. The teeth were imaged before and after irradiation using light microscopy with a flashing blue light-emitting diode (LED). An environmental scanning electron microscope imaged each tooth after irradiation. High-order super-Gaussian intensity profiles are observed at the output of a fiber coiled around a 4-in. diameter drum. Super-Gaussian beams have a more-homogenous fluence distribution than Gaussian beams and have a higher energy efficiency for selective ablation. Coaxial water-cooling does not noticeably distort the intensity distribution within 1 mm from the optical fiber. In contrast, lasers focused to a Gaussian cross section (Calculus is preferentially ablated at high fluences (> or =2 J/cm(2)); below this fluence, stalling occurs because of photo-bleaching of the calculus. Healthy dental hard tissue is not removed at fluences calculus with a frequency-doubled Ti:sapphire laser. Fluences over 2 J/cm(2) are required to remove calculus efficiently since photo-bleaching stalls calculus removal below that value.

  8. The structure and function of presynaptic endosomes

    Energy Technology Data Exchange (ETDEWEB)

    Jähne, Sebastian, E-mail: sebastian.jaehne1@stud.uni-goettingen.de [Department of Neuro- and Sensory Physiology, University of Göttingen Medical Center, Cluster of Excellence Nanoscale Microscopy and Molecular Physiology of the Brain, Humboldtallee 23, 37073 Göttingen (Germany); International Max Planck Research School for Neurosciences, 37077 Göttingen (Germany); Rizzoli, Silvio O. [Department of Neuro- and Sensory Physiology, University of Göttingen Medical Center, Cluster of Excellence Nanoscale Microscopy and Molecular Physiology of the Brain, Humboldtallee 23, 37073 Göttingen (Germany); Helm, Martin S., E-mail: martin.helm@med.uni-goettingen.de [Department of Neuro- and Sensory Physiology, University of Göttingen Medical Center, Cluster of Excellence Nanoscale Microscopy and Molecular Physiology of the Brain, Humboldtallee 23, 37073 Göttingen (Germany); International Max Planck Research School for Molecular Biology, 37077 Göttingen (Germany)

    2015-07-15

    The function of endosomes and of endosome-like structures in the presynaptic compartment is still controversial. This is in part due to the absence of a consensus on definitions and markers for these compartments. Synaptic endosomes are sometimes seen as stable organelles, permanently present in the synapse. Alternatively, they are seen as short-lived intermediates in synaptic vesicle recycling, arising from the endocytosis of large vesicles from the plasma membrane, or from homotypic fusion of small vesicles. In addition, the potential function of the endosome is largely unknown in the synapse. Some groups have proposed that the endosome is involved in the sorting of synaptic vesicle proteins, albeit others have produced data that deny this possibility. In this review, we present the existing evidence for synaptic endosomes, we discuss their potential functions, and we highlight frequent technical pitfalls in the analysis of this elusive compartment. We also sketch a roadmap to definitely determine the role of synaptic endosomes for the synaptic vesicle cycle. Finally, we propose a common definition of synaptic endosome-like structures.

  9. The structure and function of presynaptic endosomes

    International Nuclear Information System (INIS)

    Jähne, Sebastian; Rizzoli, Silvio O.; Helm, Martin S.

    2015-01-01

    The function of endosomes and of endosome-like structures in the presynaptic compartment is still controversial. This is in part due to the absence of a consensus on definitions and markers for these compartments. Synaptic endosomes are sometimes seen as stable organelles, permanently present in the synapse. Alternatively, they are seen as short-lived intermediates in synaptic vesicle recycling, arising from the endocytosis of large vesicles from the plasma membrane, or from homotypic fusion of small vesicles. In addition, the potential function of the endosome is largely unknown in the synapse. Some groups have proposed that the endosome is involved in the sorting of synaptic vesicle proteins, albeit others have produced data that deny this possibility. In this review, we present the existing evidence for synaptic endosomes, we discuss their potential functions, and we highlight frequent technical pitfalls in the analysis of this elusive compartment. We also sketch a roadmap to definitely determine the role of synaptic endosomes for the synaptic vesicle cycle. Finally, we propose a common definition of synaptic endosome-like structures

  10. Hepatitis C Virus Replication Depends on Endosomal Cholesterol Homeostasis.

    Science.gov (United States)

    Stoeck, Ina Karen; Lee, Ji-Young; Tabata, Keisuke; Romero-Brey, Inés; Paul, David; Schult, Philipp; Lohmann, Volker; Kaderali, Lars; Bartenschlager, Ralf

    2018-01-01

    Similar to other positive-strand RNA viruses, hepatitis C virus (HCV) causes massive rearrangements of intracellular membranes, resulting in a membranous web (MW) composed of predominantly double-membrane vesicles (DMVs), the presumed sites of RNA replication. DMVs are enriched for cholesterol, but mechanistic details on the source and recruitment of cholesterol to the viral replication organelle are only partially known. Here we focused on selected lipid transfer proteins implicated in direct lipid transfer at various endoplasmic reticulum (ER)-membrane contact sites. RNA interference (RNAi)-mediated knockdown identified several hitherto unknown HCV dependency factors, such as steroidogenic acute regulatory protein-related lipid transfer domain protein 3 (STARD3), oxysterol-binding protein-related protein 1A and -B (OSBPL1A and -B), and Niemann-Pick-type C1 (NPC1), all residing at late endosome and lysosome membranes and required for efficient HCV RNA replication but not for replication of the closely related dengue virus. Focusing on NPC1, we found that knockdown or pharmacological inhibition caused cholesterol entrapment in lysosomal vesicles concomitant with decreased cholesterol abundance at sites containing the viral replicase factor NS5A. In untreated HCV-infected cells, unesterified cholesterol accumulated at the perinuclear region, partially colocalizing with NS5A at DMVs, arguing for NPC1-mediated endosomal cholesterol transport to the viral replication organelle. Consistent with cholesterol being an important structural component of DMVs, reducing NPC1-dependent endosomal cholesterol transport impaired MW integrity. This suggests that HCV usurps lipid transfer proteins, such as NPC1, at ER-late endosome/lysosome membrane contact sites to recruit cholesterol to the viral replication organelle, where it contributes to MW functionality. IMPORTANCE A key feature of the replication of positive-strand RNA viruses is the rearrangement of the host cell

  11. Direct chemical-analysis of uv laser-ablation products of organic polymers by using selective ion monitoring mode in gas-chromatography mass-spectrometry

    Science.gov (United States)

    Cho, Yirang; Lee, H.W.; Fountain, S.T.; Lubman, D.M.

    1994-01-01

    Trace quantities of laser ablated organic polymers were analyzed by using commercial capillary column gas chromatography/mass spectrometry; the instrument was modified so that the laser ablation products could be introduced into the capillary column directly and the constituents of each peak in the chromatogram were identified by using a mass spectrometer. The present study takes advantage of the selective ion monitoring mode for significantly improving the sensitivity of the mass spectrometer as a detector, which is critical in analyzing the trace quantities and confirming the presence or absence of the species of interest in laser ablated polymers. The initial composition of the laser ablated polymers was obtained by using an electron impact reflectron time-of-flight mass spectrometer and the possible structure of the fragments observed in the spectra was proposed based on the structure of the polymers.

  12. Trace isotope analysis using resonance ionization mass spectrometry based on isotope selection with doppler shift of laser ablated atoms

    International Nuclear Information System (INIS)

    Higuchi, Yuki; Watanabe, Kenichi; Kawarabayashi, Jun; Iguchi, Tetsuo

    2005-01-01

    We have proposed a novel isotope selective Resonance Ionization Mass Spectroscopy (RIMS) concept, which can avoid the Doppler broadening on solid sample direct measurement based on laser ablation technique. We have succeeded in experimentally demonstrating the principle of our RIMS concept. Through comparison between the simulated and experimental results, we have validated the simulation model. It would be concluded from these results that we could achieve the isotope selectivity defined as the ratio of 41 Ca to 40 Ca sensitivity to be 4.5x10 10 by adopting the multi-step excitation scheme in the present method. As future works, we will try to experimentally perform the multi-step excitation scheme and improve the detection efficiency by modifying the ion extraction configuration. (author)

  13. Selective ablation of dental enamel and dentin using femtosecond laser pulses

    International Nuclear Information System (INIS)

    Lizarelli, R F Z; Costa, M M; Carvalho-Filho, E; Bagnato, V S; Nunes, F D

    2008-01-01

    The study of the interaction of intense laser light with matter, as well as transient response of atoms and molecules is very appropriated because of the laser energy concentration in a femtosecond optical pulses. The fundamental problem to be solved is to find tools and techniques which allow us to observe and manipulate on a femtosecond time scale the photonics events on and into the matter. Six third human extracted molars were exposed to a femtosecond Ti:Sapphire Q-switched and mode locked laser (Libra-S, Coherent, Palo Alto, CA, USA), emitting pulses with 70 fs width, radiation wavelength of 801 nm, at a constant pulse repetition rate of 1 KHz. The laser was operated at different power levels (70 to 400 mW) with constant exposition time of 10 seconds, at focused and defocused mode. Enamel and dentin surfaces were evaluated concerned ablation rate and morphological aspects under scanning electron microscopic. The results in this present experiment suggest that at the focused mode and under higher average power, enamel tissues present microcavities with higher depth and very precise edges, but, while dentin shows a larger melt-flushing, lower depth and melting and solidification aspect. In conclusion, it is possible to choose hard or soft ablation, under lower and higher average power, respectively, revealing different aspects of dental enamel and dentin, depending on the average power, fluence and distance from the focal point of the ultra-short pulse laser on the tooth surface

  14. Late endosomal cholesterol accumulation leads to impaired intra-endosomal trafficking.

    Directory of Open Access Journals (Sweden)

    Komla Sobo

    Full Text Available BACKGROUND: Pathological accumulation of cholesterol in late endosomes is observed in lysosomal storage diseases such as Niemann-Pick type C. We here analyzed the effects of cholesterol accumulation in NPC cells, or as phenocopied by the drug U18666A, on late endosomes membrane organization and dynamics. METHODOLOGY/PRINCIPAL FINDINGS: Cholesterol accumulation did not lead to an increase in the raft to non-raft membrane ratio as anticipated. Strikingly, we observed a 2-3 fold increase in the size of the compartment. Most importantly, properties and dynamics of late endosomal intralumenal vesicles were altered as revealed by reduced late endosomal vacuolation induced by the mutant pore-forming toxin ASSP, reduced intoxication by the anthrax lethal toxin and inhibition of infection by the Vesicular Stomatitis Virus. CONCLUSIONS/SIGNIFICANCE: These results suggest that back fusion of intralumenal vesicles with the limiting membrane of late endosomes is dramatically perturbed upon cholesterol accumulation.

  15. PSAPP mice exhibit regionally selective reductions in gliosis and plaque deposition in response to S100B ablation

    Directory of Open Access Journals (Sweden)

    Young Keith A

    2010-11-01

    Full Text Available Abstract Background Numerous studies have reported that increased expression of S100B, an intracellular Ca2+ receptor protein and secreted neuropeptide, exacerbates Alzheimer's disease (AD pathology. However, the ability of S100B inhibitors to prevent/reverse AD histopathology remains controversial. This study examines the effect of S100B ablation on in vivo plaque load, gliosis and dystrophic neurons. Methods Because S100B-specific inhibitors are not available, genetic ablation was used to inhibit S100B function in the PSAPP AD mouse model. The PSAPP/S100B-/- line was generated by crossing PSAPP double transgenic males with S100B-/- females and maintained as PSAPP/S100B+/- crosses. Congo red staining was used to quantify plaque load, plaque number and plaque size in 6 month old PSAPP and PSAPP/S100B-/- littermates. The microglial marker Iba1 and astrocytic marker glial fibrillary acidic protein (GFAP were used to quantify gliosis. Dystrophic neurons were detected with the phospho-tau antibody AT8. S100B immunohistochemistry was used to assess the spatial distribution of S100B in the PSAPP line. Results PSAPP/S100B-/- mice exhibited a regionally selective decrease in cortical but not hippocampal plaque load when compared to PSAPP littermates. This regionally selective reduction in plaque load was accompanied by decreases in plaque number, GFAP-positive astrocytes, Iba1-positive microglia and phospho-tau positive dystrophic neurons. These effects were not attributable to regional variability in the distribution of S100B. Hippocampal and cortical S100B immunoreactivity in PSAPP mice was associated with plaques and co-localized with astrocytes and microglia. Conclusions Collectively, these data support S100B inhibition as a novel strategy for reducing cortical plaque load, gliosis and neuronal dysfunction in AD and suggest that both extracellular as well as intracellular S100B contribute to AD histopathology.

  16. Recurrence predictive models for patients with hepatocellular carcinoma after radiofrequency ablation using support vector machines with feature selection methods.

    Science.gov (United States)

    Liang, Ja-Der; Ping, Xiao-Ou; Tseng, Yi-Ju; Huang, Guan-Tarn; Lai, Feipei; Yang, Pei-Ming

    2014-12-01

    Recurrence of hepatocellular carcinoma (HCC) is an important issue despite effective treatments with tumor eradication. Identification of patients who are at high risk for recurrence may provide more efficacious screening and detection of tumor recurrence. The aim of this study was to develop recurrence predictive models for HCC patients who received radiofrequency ablation (RFA) treatment. From January 2007 to December 2009, 83 newly diagnosed HCC patients receiving RFA as their first treatment were enrolled. Five feature selection methods including genetic algorithm (GA), simulated annealing (SA) algorithm, random forests (RF) and hybrid methods (GA+RF and SA+RF) were utilized for selecting an important subset of features from a total of 16 clinical features. These feature selection methods were combined with support vector machine (SVM) for developing predictive models with better performance. Five-fold cross-validation was used to train and test SVM models. The developed SVM-based predictive models with hybrid feature selection methods and 5-fold cross-validation had averages of the sensitivity, specificity, accuracy, positive predictive value, negative predictive value, and area under the ROC curve as 67%, 86%, 82%, 69%, 90%, and 0.69, respectively. The SVM derived predictive model can provide suggestive high-risk recurrent patients, who should be closely followed up after complete RFA treatment. Copyright © 2014 Elsevier Ireland Ltd. All rights reserved.

  17. Selective ablation of Copper-Indium-Diselenide solar cells monitored by laser-induced breakdown spectroscopy and classification methods

    Energy Technology Data Exchange (ETDEWEB)

    Diego-Vallejo, David [Technische Universität Berlin, Institute of Optics and Atomic Physics, Straße des 17, Juni 135, 10623 Berlin (Germany); Laser- und Medizin- Technologie Berlin GmbH (LMTB), Applied Laser Technology, Fabeckstr. 60-62, 14195 Berlin (Germany); Ashkenasi, David, E-mail: d.ashkenasi@lmtb.de [Laser- und Medizin- Technologie Berlin GmbH (LMTB), Applied Laser Technology, Fabeckstr. 60-62, 14195 Berlin (Germany); Lemke, Andreas [Laser- und Medizin- Technologie Berlin GmbH (LMTB), Applied Laser Technology, Fabeckstr. 60-62, 14195 Berlin (Germany); Eichler, Hans Joachim [Technische Universität Berlin, Institute of Optics and Atomic Physics, Straße des 17, Juni 135, 10623 Berlin (Germany); Laser- und Medizin- Technologie Berlin GmbH (LMTB), Applied Laser Technology, Fabeckstr. 60-62, 14195 Berlin (Germany)

    2013-09-01

    Laser-induced breakdown spectroscopy (LIBS) and two classification methods, i.e. linear correlation and artificial neural networks (ANN), are used to monitor P1, P2 and P3 scribing steps of Copper-Indium-Diselenide (CIS) solar cells. Narrow channels featuring complete removal of desired layers with minimum damage on the underlying film are expected to enhance efficiency of solar cells. The monitoring technique is intended to determine that enough material has been removed to reach the desired layer based on the analysis of plasma emission acquired during multiple pass laser scribing. When successful selective scribing is achieved, a high degree of similarity between test and reference spectra has to be identified by classification methods in order to stop the scribing procedure and avoid damaging the bottom layer. Performance of linear correlation and artificial neural networks is compared and evaluated for two spectral bandwidths. By using experimentally determined combinations of classifier and analyzed spectral band for each step, classification performance achieves errors of 7, 1 and 4% for steps P1, P2 and P3, respectively. The feasibility of using plasma emission for the supervision of processing steps of solar cell manufacturing is demonstrated. This method has the potential to be implemented as an online monitoring procedure assisting the production of solar cells. - Highlights: • LIBS and two classification methods were used to monitor CIS solar cells processing. • Selective ablation of thin-film solar cells was improved with inspection system. • Customized classification method and analyzed spectral band enhanced performance.

  18. Cell-specific cre recombinase expression allows selective ablation of glutamate receptors from mouse horizontal cells.

    Directory of Open Access Journals (Sweden)

    Sebastian Ströh

    Full Text Available In the mouse retina, horizontal cells form an electrically coupled network and provide feedback signals to photoreceptors and feedforward signals to bipolar cells. Thereby, horizontal cells contribute to gain control at the first visual synapse and to the antagonistic organization of bipolar and ganglion cell receptive fields. However, the nature of horizontal cell output remains a matter of debate, just as the exact contribution of horizontal cells to center-surround antagonism. To facilitate studying horizontal cell function, we developed a knockin mouse line which allows ablating genes exclusively in horizontal cells. This knockin line expresses a Cre recombinase under the promoter of connexin57 (Cx57, a gap junction protein only expressed in horizontal cells. Consistently, in Cx57+/Cre mice, Cre recombinase is expressed in almost all horizontal cells (>99% and no other retinal neurons. To test Cre activity, we crossbred Cx57+/Cre mice with a mouse line in which exon 11 of the coding sequence for the ionotropic glutamate receptor subunit GluA4 was flanked by two loxP sites (GluA4fl/fl. In GluA4fl/fl:Cx57+/Cre mice, GluA4 immunoreactivity was significantly reduced (∼ 50% in the outer retina where horizontal cells receive photoreceptor inputs, confirming the functionality of the Cre/loxP system. Whole-cell patch-clamp recordings from isolated horizontal cell somata showed a reduction of glutamate-induced inward currents by ∼ 75%, suggesting that the GluA4 subunit plays a major role in mediating photoreceptor inputs. The persistent current in GluA4-deficient cells is mostly driven by AMPA and to a very small extent by kainate receptors as revealed by application of the AMPA receptor antagonist GYKI52466 and concanavalin A, a potentiator of kainate receptor-mediated currents. In summary, the Cx57+/Cre mouse line provides a versatile tool for studying horizontal cell function. GluA4fl/fl:Cx57+/Cre mice, in which horizontal cells receive less

  19. Cell-Specific Cre Recombinase Expression Allows Selective Ablation of Glutamate Receptors from Mouse Horizontal Cells

    Science.gov (United States)

    Janssen-Bienhold, Ulrike; Schultz, Konrad; Cimiotti, Kerstin; Weiler, Reto; Willecke, Klaus; Dedek, Karin

    2013-01-01

    In the mouse retina, horizontal cells form an electrically coupled network and provide feedback signals to photoreceptors and feedforward signals to bipolar cells. Thereby, horizontal cells contribute to gain control at the first visual synapse and to the antagonistic organization of bipolar and ganglion cell receptive fields. However, the nature of horizontal cell output remains a matter of debate, just as the exact contribution of horizontal cells to center-surround antagonism. To facilitate studying horizontal cell function, we developed a knockin mouse line which allows ablating genes exclusively in horizontal cells. This knockin line expresses a Cre recombinase under the promoter of connexin57 (Cx57), a gap junction protein only expressed in horizontal cells. Consistently, in Cx57+/Cre mice, Cre recombinase is expressed in almost all horizontal cells (>99%) and no other retinal neurons. To test Cre activity, we crossbred Cx57+/Cre mice with a mouse line in which exon 11 of the coding sequence for the ionotropic glutamate receptor subunit GluA4 was flanked by two loxP sites (GluA4fl/fl). In GluA4fl/fl:Cx57+/Cre mice, GluA4 immunoreactivity was significantly reduced (∼50%) in the outer retina where horizontal cells receive photoreceptor inputs, confirming the functionality of the Cre/loxP system. Whole-cell patch-clamp recordings from isolated horizontal cell somata showed a reduction of glutamate-induced inward currents by ∼75%, suggesting that the GluA4 subunit plays a major role in mediating photoreceptor inputs. The persistent current in GluA4-deficient cells is mostly driven by AMPA and to a very small extent by kainate receptors as revealed by application of the AMPA receptor antagonist GYKI52466 and concanavalin A, a potentiator of kainate receptor-mediated currents. In summary, the Cx57+/Cre mouse line provides a versatile tool for studying horizontal cell function. GluA4fl/fl:Cx57+/Cre mice, in which horizontal cells receive less excitatory input

  20. Small Molecules for Early Endosome-Specific Patch Clamping.

    Science.gov (United States)

    Chen, Cheng-Chang; Butz, Elisabeth S; Chao, Yu-Kai; Grishchuk, Yulia; Becker, Lars; Heller, Stefan; Slaugenhaupt, Susan A; Biel, Martin; Wahl-Schott, Christian; Grimm, Christian

    2017-07-20

    To resolve the subcellular distribution of endolysosomal ion channels, we have established a novel experimental approach to selectively patch clamp Rab5 positive early endosomes (EE) versus Rab7/LAMP1-positive late endosomes/lysosomes (LE/LY). To functionally characterize ion channels in endolysosomal membranes with the patch-clamp technique, it is important to develop techniques to selectively enlarge the respective organelles. We found here that two small molecules, wortmannin and latrunculin B, enlarge Rab5-positive EE when combined but not Rab7-, LAMP1-, or Rab11 (RE)-positive vesicles. The two compounds act rapidly, specifically, and are readily applicable in contrast to genetic approaches or previously used compounds such as vacuolin, which enlarges EE, RE, and LE/LY. We apply this approach here to measure currents mediated by TRPML channels, in particular TRPML3, which we found to be functionally active in both EE and LE/LY in overexpressing cells as well as in endogenously expressing CD11b+ lung-tissue macrophages. Copyright © 2017 Elsevier Ltd. All rights reserved.

  1. Molecular basis for SNX-BAR-mediated assembly of distinct endosomal sorting tubules

    DEFF Research Database (Denmark)

    van Weering, Jan R.T.; Sessions, Richard B.; Traer, Colin J.

    2012-01-01

    that dimerization is achieved in part through neutralization of charged residues in the hydrophobic BAR-dimerization interface. Membrane remodelling also requires functional amphipathic helices, predicted to be present in all SNX-BARs, and the formation of high order SNX-BAR oligomers through selective 'tip...... and organizes the tubular endosomal network....

  2. Neurotrophin signaling endosomes; biogenesis, regulation, and functions

    Science.gov (United States)

    Yamashita, Naoya; Kuruvilla, Rejji

    2016-01-01

    In the nervous system, communication between neurons and their post-synaptic target cells is critical for the formation, refinement and maintenance of functional neuronal connections. Diffusible signals secreted by target tissues, exemplified by the family of neurotrophins, impinge on nerve terminals to influence diverse developmental events including neuronal survival and axonal growth. Key mechanisms of action of target-derived neurotrophins include the cell biological processes of endocytosis and retrograde trafficking of their Trk receptors from growth cones to cell bodies. In this review, we summarize the molecular mechanisms underlying this endosome-mediated signaling, focusing on the instructive role of neurotrophin signaling itself in directing its own trafficking. Recent studies have linked impaired neurotrophin trafficking to neurodevelopmental disorders, highlighting the relevance of neurotrophin endosomes in human health. PMID:27327126

  3. Conformational biosensors reveal GPCR signalling from endosomes

    DEFF Research Database (Denmark)

    Irannejad, R; Tomshine, Jin C; Tomshine, Jon R

    2013-01-01

    A long-held tenet of molecular pharmacology is that canonical signal transduction mediated by G-protein-coupled receptor (GPCR) coupling to heterotrimeric G proteins is confined to the plasma membrane. Evidence supporting this traditional view is based on analytical methods that provide limited...... or no subcellular resolution. It has been subsequently proposed that signalling by internalized GPCRs is restricted to G-protein-independent mechanisms such as scaffolding by arrestins, or GPCR activation elicits a discrete form of persistent G protein signalling, or that internalized GPCRs can indeed contribute...... in the early endosome membrane, and that internalized receptors contribute to the overall cellular cyclic AMP response within several minutes after agonist application. These findings provide direct support for the hypothesis that canonical GPCR signalling occurs from endosomes as well as the plasma membrane...

  4. Selective ablation of dental calculus with a frequency-doubled Alexandrite laser

    Science.gov (United States)

    Rechmann, Peter; Hennig, Thomas

    1996-01-01

    The aim of the study was the selective removal of dental calculus by means of pulsed lasers. In a first approach the optical characteristics of subgingival calculus were calculated using fluorescence emission spectroscopy (excitation laser: N2-laser, wavelength 337 nm, pulse duration 4 ns). Subgingival calculus seems to absorb highly in the ultraviolet spectral region up to 420 nm. According to these measurements a frequency doubled Alexandrite-laser (wavelength 377 nm, pulse duration 100 ns, repetition rate 110 Hz) was used to irradiate calculus located on enamel, at the cementum enamel junction and on the root surface (located on dentin or on cementum). Irradiation was performed perpendicular to the root surface with a laser fluence of 1 Jcm-2. During the irradiation procedure an effective water cooling-system was engaged. Histological investigations were done on undecalcified sections. As a result, engaging low fluences allows a fast and strictly selective removal of subgingival calculus. Even more the investigations revealed that supragingival calculus can be removed in a strictly selective manner engaging a frequency doubled Alexandrite-laser. No adverse side effects to the surrounding tissues could be found.

  5. The Endosome Localized Arf-GAP AGAP1 Modulates Dendritic Spine Morphology Downstream of the Neurodevelopmental Disorder Factor Dysbindin

    Directory of Open Access Journals (Sweden)

    Miranda Arnold

    2016-09-01

    Full Text Available AGAP1 is an Arf1 GTPase activating protein that interacts with the vesicle-associated protein complexes adaptor protein 3 (AP-3 and Biogenesis of Lysosome Related Organelles Complex-1 (BLOC-1. Overexpression of AGAP1 in non-neuronal cells results in an accumulation of endosomal cargoes, which suggests a role in endosome-dependent traffic. In addition, AGAP1 is a candidate susceptibility gene for two neurodevelopmental disorders, autism spectrum disorder (ASD and schizophrenia (SZ; yet its localization and function in neurons have not been described. Here, we describe that AGAP1 localizes to axons, dendrites, dendritic spines, and synapses, colocalizing preferentially with markers of early and recycling endosomes. Functional studies reveal overexpression and down-regulation of AGAP1 affects both neuronal endosomal trafficking and dendritic spine morphology, supporting a role for AGAP1 in the recycling endosomal trafficking involved in their morphogenesis. Finally, we determined the sensitivity of AGAP1 expression to mutations in the DTNBP1 gene, which is associated with neurodevelopmental disorder, and found that AGAP1 mRNA and protein levels are selectively reduced in the null allele of the mouse orthologue of DTNBP1. We postulate that endosomal trafficking contributes to the pathogenesis of neurodevelopmental disorders affecting dendritic spine morphology, and thus excitatory synapse structure and function.

  6. Immunomodulation-accelerated neuronal regeneration following selective rod photoreceptor cell ablation in the zebrafish retina.

    Science.gov (United States)

    White, David T; Sengupta, Sumitra; Saxena, Meera T; Xu, Qingguo; Hanes, Justin; Ding, Ding; Ji, Hongkai; Mumm, Jeff S

    2017-05-02

    Müller glia (MG) function as inducible retinal stem cells in zebrafish, completely repairing the eye after damage. The innate immune system has recently been shown to promote tissue regeneration in which classic wound-healing responses predominate. However, regulatory roles for leukocytes during cellular regeneration-i.e., selective cell-loss paradigms akin to degenerative disease-are less well defined. To investigate possible roles innate immune cells play during retinal cell regeneration, we used intravital microscopy to visualize neutrophil, macrophage, and retinal microglia responses to induced rod photoreceptor apoptosis. Neutrophils displayed no reactivity to rod cell loss. Peripheral macrophage cells responded to rod cell loss, as evidenced by morphological transitions and increased migration, but did not enter the retina. Retinal microglia displayed multiple hallmarks of immune cell activation: increased migration, translocation to the photoreceptor cell layer, proliferation, and phagocytosis of dying cells. To test function during rod cell regeneration, we coablated microglia and rod cells or applied immune suppression and quantified the kinetics of ( i ) rod cell clearance, ( ii ) MG/progenitor cell proliferation, and ( iii ) rod cell replacement. Coablation and immune suppressants applied before cell loss caused delays in MG/progenitor proliferation rates and slowed the rate of rod cell replacement. Conversely, immune suppressants applied after cell loss had been initiated led to accelerated photoreceptor regeneration kinetics, possibly by promoting rapid resolution of an acute immune response. Our findings suggest that microglia control MG responsiveness to photoreceptor loss and support the development of immune-targeted therapeutic strategies for reversing cell loss associated with degenerative retinal conditions.

  7. Cardiac ablation

    Directory of Open Access Journals (Sweden)

    Kelly Ratheal

    2016-01-01

    Full Text Available Cardiac ablation is a procedure that uses either radiofrequency or cryothermal energy to destroy cells in the heart to terminate and/or prevent arrhythmias. The indications for cardiac catheter ablation include refractory, symptomatic arrhythmias, with more specific guidelines for atrial fibrillation in particular. The ablation procedure itself involves mapping the arrhythmia and destruction of the aberrant pathway in an effort to permanently prevent the arrhythmia. There are many types of arrhythmias, and they require individualized approaches to ablation based on their innately different electrical pathways. Ablation of arrhythmias, such as Wolff-Parkinson-White syndrome, AV nodal reentrant tachycardia, and atrial-fibrillation, is discussed in this review. Ablation has a high success rate overall and minimal complication rates, leading to improved quality of life in many patients.

  8. Selective ablation of the androgen receptor in mouse sertoli cells affects sertoli cell maturation, barrier formation and cytoskeletal development.

    Directory of Open Access Journals (Sweden)

    Ariane Willems

    2010-11-01

    Full Text Available The observation that mice with a selective ablation of the androgen receptor (AR in Sertoli cells (SC (SCARKO mice display a complete block in meiosis supports the contention that SC play a pivotal role in the control of germ cell development by androgens. To delineate the physiological and molecular mechanism responsible for this control, we compared tubular development in pubertal SCARKO mice and littermate controls. Particular attention was paid to differences in SC maturation, SC barrier formation and cytoskeletal organization and to the molecular mediators potentially involved. Functional analysis of SC barrier development by hypertonic perfusion and lanthanum permeation techniques and immunohistochemical analysis of junction formation showed that SCARKO mice still attempt to produce a barrier separating basal and adluminal compartment but that barrier formation is delayed and defective. Defective barrier formation was accompanied by disturbances in SC nuclear maturation (immature shape, absence of prominent, tripartite nucleoli and SC polarization (aberrant positioning of SC nuclei and cytoskeletal elements such as vimentin. Quantitative RT-PCR was used to study the transcript levels of genes potentially related to the described phenomena between day 8 and 35. Differences in the expression of SC genes known to play a role in junction formation could be shown from day 8 for Cldn11, from day 15 for Cldn3 and Espn, from day 20 for Cdh2 and Jam3 and from day 35 for ZO-1. Marked differences were also noted in the transcript levels of several genes that are also related to cell adhesion and cytoskeletal dynamics but that have not yet been studied in SC (Actn3, Ank3, Anxa9, Scin, Emb, Mpzl2. It is concluded that absence of a functional AR in SC impedes the remodeling of testicular tubules expected at the onset of spermatogenesis and interferes with the creation of the specific environment needed for germ cell development.

  9. Ablative skin resurfacing.

    Science.gov (United States)

    Agrawal, Nidhi; Smith, Greg; Heffelfinger, Ryan

    2014-02-01

    Ablative laser resurfacing has evolved as a safe and effective treatment for skin rejuvenation. Although traditional lasers were associated with significant thermal damage and lengthy recovery, advances in laser technology have improved safety profiles and reduced social downtime. CO2 lasers remain the gold standard of treatment, and fractional ablative devices capable of achieving remarkable clinical improvement with fewer side effects and shorter recovery times have made it a more practical option for patients. Although ablative resurfacing has become safer, careful patient selection and choice of suitable laser parameters are essential to minimize complications and optimize outcomes. This article describes the current modalities used in ablative laser skin resurfacing and examines their efficacy, indications, and possible side effects. Thieme Medical Publishers 333 Seventh Avenue, New York, NY 10001, USA.

  10. Molecular assemblies and membrane domains in multivesicular endosome dynamics

    International Nuclear Information System (INIS)

    Falguieres, Thomas; Luyet, Pierre-Philippe; Gruenberg, Jean

    2009-01-01

    Along the degradation pathway, endosomes exhibit a characteristic multivesicular organization, resulting from the budding of vesicles into the endosomal lumen. After endocytosis and transport to early endosomes, activated signaling receptors are incorporated into these intralumenal vesicles through the action of the ESCRT machinery, a process that contributes to terminate signaling. Then, the vesicles and their protein cargo are further transported towards lysosomes for degradation. Evidence also shows that intralumenal vesicles can undergo 'back-fusion' with the late endosome limiting membrane, a route exploited by some pathogens and presumably followed by proteins and lipids that need to be recycled from within the endosomal lumen. This process depends on the late endosomal lipid lysobisphosphatidic acid and its putative effector Alix/AIP1, and is presumably coupled to the invagination of the endosomal limiting membrane at the molecular level via ESCRT proteins. In this review, we discuss the intra-endosomal transport routes in mammalian cells, and in particular the different mechanisms involved in membrane invagination, vesicle formation and fusion in a space inaccessible to proteins known to control intracellular membrane traffic.

  11. Power Laser Ablation Symposia

    CERN Document Server

    Phipps, Claude

    2007-01-01

    Laser ablation describes the interaction of intense optical fields with matter, in which atoms are selectively driven off by thermal or nonthermal mechanisms. The field of laser ablation physics is advancing so rapidly that its principal results are seen only in specialized journals and conferences. This is the first book that combines the most recent results in this rapidly advancing field with authoritative treatment of laser ablation and its applications, including the physics of high-power laser-matter interaction. Many practical applications exist, ranging from inertial confinement fusion to propulsion of aerostats for pollution monitoring to laser ignition of hypersonic engines to laser cleaning nanoscale contaminants in high-volume computer hard drive manufacture to direct observation of the electronic or dissociative states in atoms and molecules, to studying the properties of materials during 200kbar shocks developed in 200fs. Selecting topics which are representative of such a broad field is difficu...

  12. Lysosomal and endosomal heterogeneity in the liver: A comparison of the intracellular pathways of endocytosis in rat liver cells

    International Nuclear Information System (INIS)

    Kindberg, G.M.; Tolleshaug, H.; Gjoen, T.; Berg, T.

    1991-01-01

    Air-filled albumin microspheres, asialoorosomucoid and formaldehyde-treated serum albumin are selectively taken up by endocytosis in rat liver Kupffer cells, parenchymal cells and endothelial cells, respectively. Intracellular transport and degradation of endocytosed material were studied by subcellular fractionation in sucrose and Nycodenz gradients after intravenous injection of the ligand. By using ligands labeled with 125I-tyramine-cellobiose, the subcellular distribution of labeled degradation products can be studied because they are trapped at the site of formation. The results show that the kinetics of intracellular transport are different in hepatic parenchymal, endothelial and Kupffer cells. In endothelial cells, the ligand is associated with two types of endosomes during the first minutes after internalization and then is transferred rapidly to the lysosomes. In parenchymal cells, 125I-tyramine-cellobiose-asialoorosomucoid was located in a relatively slowly sedimenting vesicle during the first minute after internalization and subsequently in denser endosomes. Degradation of 125I-tyramine-cellobiose-asialoorosomucoid in parenchymal cells started later than that of 125I-tyramine-cellobiose-formaldehyde-treated serum albumin in endothelial cells. Furthermore, the ligand seemed to be transferred relatively slowly from endosomes to lysosomes, and most of the undegraded ligand was in the endosomes. The rate-limiting step of proteolysis in parenchymal cells is probably the transport from endosomes to lysosomes. In Kupffer cells, most 125I-tyramine-cellobiose-microspheres are found as undegraded material in very dense endosomes up to 3 hr after injection. After 20 hr, most of the ligand is degraded in lysosomes distributed at a lower density than the endosomes in Nycodenz and sucrose gradients

  13. Radiofrequency ablation of pulmonary tumors

    Energy Technology Data Exchange (ETDEWEB)

    Crocetti, Laura, E-mail: l.crocetti@med.unipi.i [Division of Diagnostic Imaging and Intervention, Department of Liver Transplants, Hepatology and Infectious Diseases, Pisa University School of Medicine (Italy); Lencioni, Riccardo [Division of Diagnostic Imaging and Intervention, Department of Liver Transplants, Hepatology and Infectious Diseases, Pisa University School of Medicine (Italy)

    2010-07-15

    The development of image-guided percutaneous techniques for local tumor ablation has been one of the major advances in the treatment of solid tumors. Among these methods, radiofrequency (RF) ablation is currently established as the primary ablative modality at most institutions. RF ablation is accepted as the best therapeutic choice for patients with early-stage hepatocellular carcinoma when liver transplantation or surgical resection are not suitable options and is considered as a viable alternate to surgery for inoperable patients with limited hepatic metastatic disease, especially from colorectal cancer. Recently, RF ablation has been demonstrated to be a safe and valuable treatment option for patients with unresectable or medically inoperable lung malignancies. Resection should remain the standard therapy for non-small cell lung cancer (NSCLC) but RF ablation may be better than conventional external-beam radiation for the treatment of the high-risk individual with NSCLC. Initial favourable outcomes encourage combining radiotherapy and RF ablation, especially for treating larger tumors. In the setting of colorectal cancer lung metastases, survival rates provided by RF ablation in selected patients, are substantially higher than those obtained with any chemotherapy regimens and provide indirect evidence that RF ablation therapy improves survival in patients with limited lung metastatic disease.

  14. Positioning of AMPA Receptor-Containing Endosomes Regulates Synapse Architecture

    Directory of Open Access Journals (Sweden)

    Marta Esteves da Silva

    2015-11-01

    Full Text Available Lateral diffusion in the membrane and endosomal trafficking both contribute to the addition and removal of AMPA receptors (AMPARs at postsynaptic sites. However, the spatial coordination between these mechanisms has remained unclear, because little is known about the dynamics of AMPAR-containing endosomes. In addition, how the positioning of AMPAR-containing endosomes affects synapse organization and functioning has never been directly explored. Here, we used live-cell imaging in hippocampal neuron cultures to show that intracellular AMPARs are transported in Rab11-positive recycling endosomes, which frequently enter dendritic spines and depend on the microtubule and actin cytoskeleton. By using chemically induced dimerization systems to recruit kinesin (KIF1C or myosin (MyosinV/VI motors to Rab11-positive recycling endosomes, we controlled their trafficking and found that induced removal of recycling endosomes from spines decreases surface AMPAR expression and PSD-95 clusters at synapses. Our data suggest a mechanistic link between endosome positioning and postsynaptic structure and composition.

  15. Recycling endosomes in human cytotoxic T lymphocytes constitute an auxiliary intracellular trafficking pathway for newly synthesized perforin

    Science.gov (United States)

    Lesteberg, Kelsey E.; Orange, Jordan S.; Makedonas, George

    2018-01-01

    Background Although cytotoxic T lymphocytes (CTLs) store perforin within cytoplasmic secretory granules for immediate use, perforin is synthesized anew within hours of TCR stimulation. Previously, we observed new perforin protein at an immunologic synapse independent of secretory lysosomes; herein we aimed to determine how new perforin transits to the synapse if not via lytic granules. Results We analyzed antigen-specific human CTLs via imaging flow cytometry and high-resolution confocal microscopy, with attention to intracellular trafficking components and new perforin. The recycling endosome compartments identified by rab8, rab11a, rab4, and rab37 co-localized with new perforin, as well as the SNAREs vti1b and VAMP4. After ablating the function of the recycling endosome pathway, we observed a relative accumulation of new perforin in rab8 vesicles. Conclusions The recycling endosome pathway may serve as an auxiliary intracellular route for the delivery of new perforin to an immunologic synapse in order to perpetuate a cytotoxic response. PMID:28822075

  16. Resonance absorption spectroscopy for laser-ablated lanthanide atom. (1) Optimized experimental conditions for isotope-selective absorption of gadolinium (Contract research)

    International Nuclear Information System (INIS)

    Miyabe, Masabumi; Oba, Masaki; Iimura, Hideki; Akaoka, Katsuaki; Maruyama, Yoichiro; Wakaida, Ikuo; Watanabe, Kazuo

    2008-06-01

    For remote isotope analysis of low-decontaminated TRU fuel, we are developing an analytical technique on the basis of the resonance absorption spectroscopy for the laser-ablation plume. To improve isotopic selectivity and detection sensitivity of this technique, we measured absorption spectra of Gd atom with various plume production conditions (ablation laser intensity, ambient gas and its pressure) and observation conditions (transition, probe height from sample, observation timing). As a result, high resolution spectrum was obtained from the observation of slow component of the plume produced under low-pressure rare-gas ambient. The observed narrowest linewidth of about 0.85GHz was found to be close to the Doppler width estimated for Gd atom of room temperature. Furthermore, relaxation rate of higher meta-stable state was found to be higher than that of ground state, suggesting that use of the transition arising from ground state or lower meta-stable state is preferable for highly sensitive isotope analysis. (author)

  17. Heterotrimeric G protein subunits are located on rat liver endosomes

    Directory of Open Access Journals (Sweden)

    Van Dyke Rebecca W

    2004-01-01

    Full Text Available Abstract Background Rat liver endosomes contain activated insulin receptors and downstream signal transduction molecules. We undertook these studies to determine whether endosomes also contain heterotrimeric G proteins that may be involved in signal transduction from G protein-coupled receptors. Results By Western blotting Gsα, Giα1,2, Giα3 and Gβ were enriched in both canalicular (CM and basolateral (BLM membranes but also readily detectable on three types of purified rat liver endosomes in the order recycling receptor compartment (RRC > compartment for uncoupling of receptor and ligand (CURL > multivesicular bodies (MVB >> purified secondary lysosomes. Western blotting with antibodies to Na, K-ATPase and to other proteins associated with plasma membranes and intracellular organelles indicated this was not due to contamination of endosome preparations by CM or BLM. Adenylate cyclase (AC was also identified on purified CM, BLM, RRC, CURL and MVB. Percoll gradient fractionation of liver postnuclear supernatants demonstrated co-occurrence of endosomes and heterotrimeric G protein subunits in fractions with little plasma membrane markers. By confocal microscopy, punctate staining for Gsα, Giα3 and Gβ corresponded to punctate areas of endocytosed Texas red-dextran in hepatocytes from control and cholera toxin-treated livers. Conclusion We conclude that heterotrimeric G protein subunits as well as AC likely traffic into hepatocytes on endosome membranes, possibly generating downstream signals spatially separate from signalling generated at the plasma membrane, analogous to the role(s of internalized insulin receptors.

  18. An innovative method to selectively ablate cholesterol esters from an arteriosclerotic region of the arterial wall using a free-electron laser

    International Nuclear Information System (INIS)

    Awazu, K.; Nagai, A.; Tomimasu, T.; Aizawa, K.

    1998-01-01

    The selective removal of cholesterol esters accumulated in the intra-cellular or extra-cellular spaces has clinical significance. In the present work we investigate the removal of cholesterol esters by using a free electron laser (FEL) in an arteriosclerotic region. Thin films of cholesteryl oleate and albumin, and the cross-section of a rabbit artery were placed on an inverted microscope stage and the changes caused by the FEL irradiation of 5.75 and 6.1 μm with 1.5-3 mW in average were monitored continuously by a CCD camera in real time. FEL irradiation at a wavelength of 5.75 μm, which is a stretching vibrational mode of the ester, was able to ablate cholesterol esters without affecting albumin and that it can also remove cholesterol esters from rabbits' arteriosclerotic arterial walls. (Copyright (c) 1998 Elsevier Science B.V., Amsterdam. All rights reserved.)

  19. Cholesterol transfer at endosomal-organelle membrane contact sites.

    Science.gov (United States)

    Ridgway, Neale D; Zhao, Kexin

    2018-06-01

    Cholesterol is delivered to the limiting membrane of late endosomes by Niemann-Pick Type C1 and C2 proteins. This review summarizes recent evidence that cholesterol transfer from endosomes to the endoplasmic reticulum and other organelles is mediated by lipid-binding proteins that localize to membrane contact sites (MCS). LDL-cholesterol in the late endosomal/lysosomes is exported to the plasma membrane, where most cholesterol resides, and the endoplasmic reticulum, which harbors the regulatory complexes and enzymes that control the synthesis and esterification of cholesterol. A major advance in dissecting these cholesterol transport pathways was identification of frequent and dynamic MCS between endosomes and the endoplasmic reticulum, peroxisomes and plasma membrane. Positioned at these MCS are members of the oxysterol-binding protein (OSBP) and steroidogenic acute regulatory protein-related lipid-transfer family of lipid transfer proteins that bridge the opposing membranes and directly or indirectly mediate cholesterol transfer. OSBP-related protein 1L (ORP1L), ORP5 and ORP6 mediate cholesterol transfer to the endoplasmic reticulum that regulates cholesterol homeostasis. ORP1L and STARD3 also move cholesterol from the endoplasmic reticulum-to-late endosomal/lysosomes under low-cholesterol conditions to facilitate intraluminal vesicle formation. Cholesterol transport also occurs at MCS with peroxisomes and possibly the plasma membrane. Frequent contacts between organelles and the endo-lysosomal vesicles are sites for bidirectional transfer of cholesterol.

  20. Apolipoprotein E Regulates Amyloid Formation within Endosomes of Pigment Cells

    Directory of Open Access Journals (Sweden)

    Guillaume van Niel

    2015-10-01

    Full Text Available Accumulation of toxic amyloid oligomers is a key feature in the pathogenesis of amyloid-related diseases. Formation of mature amyloid fibrils is one defense mechanism to neutralize toxic prefibrillar oligomers. This mechanism is notably influenced by apolipoprotein E variants. Cells that produce mature amyloid fibrils to serve physiological functions must exploit specific mechanisms to avoid potential accumulation of toxic species. Pigment cells have tuned their endosomes to maximize the formation of functional amyloid from the protein PMEL. Here, we show that ApoE is associated with intraluminal vesicles (ILV within endosomes and remain associated with ILVs when they are secreted as exosomes. ApoE functions in the ESCRT-independent sorting mechanism of PMEL onto ILVs and regulates the endosomal formation of PMEL amyloid fibrils in vitro and in vivo. This process secures the physiological formation of amyloid fibrils by exploiting ILVs as amyloid nucleating platforms.

  1. Cryoballoon Ablation for Atrial Fibrillation

    Directory of Open Access Journals (Sweden)

    Jason G. Andrade, MD

    2012-03-01

    Full Text Available Focal point-by-point radiofrequency catheter ablation has shown considerable success in the treatment of paroxysmal atrial fibrillation. However, it is not without limitations. Recent clinical and preclinical studies have demonstrated that cryothermal ablation using a balloon catheter (Artic Front©, Medtronic CryoCath LP provides an effective alternative strategy to treating atrial fibrillation. The objective of this article is to review efficacy and safety data surrounding cryoballoon ablation for paroxysmal and persistent atrial fibrillation. In addition, a practical step-by-step approach to cryoballoon ablation is presented, while highlighting relevant literature regarding: 1 the rationale for adjunctive imaging, 2 selection of an appropriate cryoballoon size, 3 predictors of efficacy, 4 advanced trouble-shooting techniques, and 5 strategies to reduce procedural complications, such as phrenic nerve palsy.

  2. Rapid endosomal escape of prickly nanodiamonds: implications for gene delivery

    Science.gov (United States)

    Chu, Zhiqin; Miu, Kaikei; Lung, Pingsai; Zhang, Silu; Zhao, Saisai; Chang, Huan-Cheng; Lin, Ge; Li, Quan

    2015-06-01

    The prickly nanodiamonds easily entered cells via endocytosis followed by unique intracellular translocation characteristics—quick endosomal escape followed by stable residence in cytoplasm. Endosomal membrane rupturing is identified as the major route of nanodiamonds’ escaping the vesicle confinement and to the cytoplasm. Little cytotoxicity is observed to associate with the nanodiamonds’ cytosolic release. Such features enable its application for gene delivery, which requires both effective cellular uptake and cytosolic release of the gene. Taking green fluorescent protein gene as an example, we demonstrate the successful cytosolic delivery and expression of such a gene using the prickly nanodiamonds as carrier.

  3. Rapid endosomal escape of prickly nanodiamonds: implications for gene delivery

    Science.gov (United States)

    Chu, Zhiqin; Miu, Kaikei; Lung, Pingsai; Zhang, Silu; Zhao, Saisai; Chang, Huan-Cheng; Lin, Ge; Li, Quan

    2015-01-01

    The prickly nanodiamonds easily entered cells via endocytosis followed by unique intracellular translocation characteristics—quick endosomal escape followed by stable residence in cytoplasm. Endosomal membrane rupturing is identified as the major route of nanodiamonds’ escaping the vesicle confinement and to the cytoplasm. Little cytotoxicity is observed to associate with the nanodiamonds’ cytosolic release. Such features enable its application for gene delivery, which requires both effective cellular uptake and cytosolic release of the gene. Taking green fluorescent protein gene as an example, we demonstrate the successful cytosolic delivery and expression of such a gene using the prickly nanodiamonds as carrier. PMID:26123532

  4. Rapid endosomal escape of prickly nanodiamonds: implications for gene delivery.

    Science.gov (United States)

    Chu, Zhiqin; Miu, Kaikei; Lung, Pingsai; Zhang, Silu; Zhao, Saisai; Chang, Huan-Cheng; Lin, Ge; Li, Quan

    2015-06-30

    The prickly nanodiamonds easily entered cells via endocytosis followed by unique intracellular translocation characteristics—quick endosomal escape followed by stable residence in cytoplasm. Endosomal membrane rupturing is identified as the major route of nanodiamonds' escaping the vesicle confinement and to the cytoplasm. Little cytotoxicity is observed to associate with the nanodiamonds' cytosolic release. Such features enable its application for gene delivery, which requires both effective cellular uptake and cytosolic release of the gene. Taking green fluorescent protein gene as an example, we demonstrate the successful cytosolic delivery and expression of such a gene using the prickly nanodiamonds as carrier.

  5. Ablative thermal protection systems

    International Nuclear Information System (INIS)

    Vaniman, J.; Fisher, R.; Wojciechowski, C.; Dean, W.

    1983-01-01

    The procedures used to establish the TPS (thermal protection system) design of the SRB (solid rocket booster) element of the Space Shuttle vehicle are discussed. A final evaluation of the adequacy of this design will be made from data obtained from the first five Shuttle flights. Temperature sensors installed at selected locations on the SRB structure covered by the TPS give information as a function of time throughout the flight. Anomalies are to be investigated and computer design thermal models adjusted if required. In addition, the actual TPS ablator material loss is to be measured after each flight and compared with analytically determined losses. The analytical methods of predicting ablator performance are surveyed. 5 references

  6. Radiofrequency Ablation of Lung Tumors

    Science.gov (United States)

    ... News Physician Resources Professions Site Index A-Z Radiofrequency Ablation (RFA) / Microwave Ablation (MWA) of Lung Tumors ... and Microwave Ablation of Lung Tumors? What are Radiofrequency and Microwave Ablation of Lung Tumors? Radiofrequency ablation, ...

  7. Biomechanics and Thermodynamics of Nanoparticle Interactions with Plasma and Endosomal Membrane Lipids in Cellular Uptake and Endosomal Escape

    Science.gov (United States)

    2015-01-01

    To be effective for cytoplasmic delivery of therapeutics, nanoparticles (NPs) taken up via endocytic pathways must efficiently transport across the cell membrane and subsequently escape from the secondary endosomes. We hypothesized that the biomechanical and thermodynamic interactions of NPs with plasma and endosomal membrane lipids are involved in these processes. Using model plasma and endosomal lipid membranes, we compared the interactions of cationic NPs composed of poly(d,l-lactide-co-glycolide) modified with the dichain surfactant didodecyldimethylammonium bromide (DMAB) or the single-chain surfactant cetyltrimethylammonium bromide (CTAB) vs anionic unmodified NPs of similar size. We validated our hypothesis in doxorubicin-sensitive (MCF-7, with relatively fluid membranes) and resistant breast cancer cells (MCF-7/ADR, with rigid membranes). Despite their cationic surface charges, DMAB- and CTAB-modified NPs showed different patterns of biophysical interaction: DMAB-modified NPs induced bending of the model plasma membrane, whereas CTAB-modified NPs condensed the membrane, thereby resisted bending. Unmodified NPs showed no effects on bending. DMAB-modified NPs also induced thermodynamic instability of the model endosomal membrane, whereas CTAB-modified and unmodified NPs had no effect. Since bending of the plasma membrane and destabilization of the endosomal membrane are critical biophysical processes in NP cellular uptake and endosomal escape, respectively, we tested these NPs for cellular uptake and drug efficacy. Confocal imaging showed that in both sensitive and resistant cells DMAB-modified NPs exhibited greater cellular uptake and escape from endosomes than CTAB-modified or unmodified NPs. Further, paclitaxel-loaded DMAB-modified NPs induced greater cytotoxicity even in resistant cells than CTAB-modified or unmodified NPs or drug in solution, demonstrating the potential of DMAB-modified NPs to overcome the transport barrier in resistant cells. In

  8. Characterization of tracked radiofrequency ablation in phantom

    International Nuclear Information System (INIS)

    Chen, Chun-Cheng R.; Miga, Michael I.; Galloway, Robert L.

    2007-01-01

    In radiofrequency ablation (RFA), successful therapy requires accurate, image-guided placement of the ablation device in a location selected by a predictive treatment plan. Current planning methods rely on geometric models of ablations that are not sensitive to underlying physical processes in RFA. Implementing plans based on computational models of RFA with image-guided techniques, however, has not been well characterized. To study the use of computational models of RFA in planning needle placement, this work compared ablations performed with an optically tracked RFA device with corresponding models of the ablations. The calibration of the tracked device allowed the positions of distal features of the device, particularly the tips of the needle electrodes, to be determined to within 1.4±0.6 mm of uncertainty. Ablations were then performed using the tracked device in a phantom system based on an agarose-albumin mixture. Images of the sliced phantom obtained from the ablation experiments were then compared with the predictions of a bioheat transfer model of RFA, which used the positional data of the tracked device obtained during ablation. The model was demonstrated to predict 90% of imaged pixels classified as being ablated. The discrepancies between model predictions and observations were analyzed and attributed to needle tracking inaccuracy as well as to uncertainties in model parameters. The results suggest the feasibility of using finite element modeling to plan ablations with predictable outcomes when implemented using tracked RFA

  9. The p25 Subunit of the Dynactin Complex is Required for Dynein-Early Endosome Interaction

    Science.gov (United States)

    2011-01-01

    early endosome movement. In filamentous hyphae , dynein powers the minus end–directed movement of early endosomes (Steinberg and Schuster 2011...observed in time-lapse sequences (Video 1; Abenza et al., 2009). In still images, early endosomes were seen to distribute along the hyphae (Fig. 2 A...nuclear distribution along elongated hyphae and also for the microtubule minus end–directed movement of early endosomes away from the tip (Morris

  10. Structural Basis for Endosomal Targeting by the Bro1 Domain

    Science.gov (United States)

    Kim, Jaewon; Sitaraman, Sujatha; Hierro, Aitor; Beach, Bridgette M.; Odorizzi, Greg; Hurley, James H.

    2010-01-01

    Summary Proteins delivered to the lysosome or the yeast vacuole via late endosomes are sorted by the ESCRT complexes and by associated proteins, including Alix and its yeast homolog Bro1. Alix, Bro1, and several other late endosomal proteins share a conserved 160 residue Bro1 domain whose boundaries, structure, and function have not been characterized. The crystal structure of the Bro1 domain of Bro1 reveals a folded core of 367 residues. The extended Bro1 domain is necessary and sufficient for binding to the ESCRT-III subunit Snf7 and for the recruitment of Bro1 to late endosomes. The structure resembles a boomerang with its concave face filled in and contains a triple tetratricopeptide repeat domain as a substructure. Snf7 binds to a conserved hydrophobic patch on Bro1 that is required for protein complex formation and for the protein-sorting function of Bro1. These results define a conserved mechanism whereby Bro1 domain-containing proteins are targeted to endosomes by Snf7 and its orthologs. PMID:15935782

  11. Characterization of the Mammalian CORVET and HOPS Complexes and Their Modular Restructuring for Endosome Specificity

    NARCIS (Netherlands)

    van der Kant, Rik; Jonker, Caspar T. H.; Wijdeven, Ruud H.; Bakker, Jeroen; Janssen, Lennert; Klumperman, Judith; Neefjes, Jacques

    2015-01-01

    Trafficking of cargo through the endosomal system depends on endosomal fusion events mediated by SNARE proteins, Rab-GTPases, and multisubunit tethering complexes. The CORVET and HOPS tethering complexes, respectively, regulate early and late endosomal tethering and have been characterized in detail

  12. Pellet ablation and ablation model development

    International Nuclear Information System (INIS)

    Houlberg, W.A.

    1989-01-01

    A broad survey of pellet ablation is given, based primarily on information presented at this meeting. The implications of various experimental observations for ablation theory are derived from qualitative arguments of the physics involved. The major elements of a more complete ablation theory are then outlined in terms of these observations. This is followed by a few suggestions on improving the connections between theory and experimental results through examination of ablation data. Although this is a rather aggressive undertaking for such a brief (and undoubtedly incomplete) assessment, some of the discussion may help us advance the understanding of pellet ablation. 17 refs

  13. Adiponectin release and insulin receptor targeting share trans-Golgi-dependent endosomal trafficking routes

    Directory of Open Access Journals (Sweden)

    Maria Rödiger

    2018-02-01

    Full Text Available Objective: Intracellular vesicle trafficking maintains cellular structures and functions. The assembly of cargo-laden vesicles at the trans-Golgi network is initiated by the ARF family of small GTPases. Here, we demonstrate the role of the trans-Golgi localized monomeric GTPase ARFRP1 in endosomal-mediated vesicle trafficking of mature adipocytes. Methods: Control (Arfrp1flox/flox and inducible fat-specific Arfrp1 knockout (Arfrp1iAT−/− mice were metabolically characterized. In vitro experiments on mature 3T3-L1 cells and primary mouse adipocytes were conducted to validate the impact of ARFRP1 on localization of adiponectin and the insulin receptor. Finally, secretion and transferrin-based uptake and recycling assays were performed with HeLa and HeLa M-C1 cells. Results: We identified the ARFRP1-based sorting machinery to be involved in vesicle trafficking relying on the endosomal compartment for cell surface delivery. Secretion of adiponectin from fat depots was selectively reduced in Arfrp1iAT−/− mice, and Arfrp1-depleted 3T3-L1 adipocytes revealed an accumulation of adiponectin in Rab11-positive endosomes. Plasma adiponectin deficiency of Arfrp1iAT−/− mice resulted in deteriorated hepatic insulin sensitivity, increased gluconeogenesis and elevated fasting blood glucose levels. Additionally, the insulin receptor, undergoing endocytic recycling after ligand binding, was less abundant at the plasma membrane of adipocytes lacking Arfrp1. This had detrimental effects on adipose insulin signaling, followed by insufficient suppression of basal lipolytic activity and impaired adipose tissue expansion. Conclusions: Our findings suggest that adiponectin secretion and insulin receptor surface targeting utilize the same post-Golgi trafficking pathways that are essential for an appropriate systemic insulin sensitivity and glucose homeostasis. Keywords: Adiponectin, ARFRP1, Exocytosis, Insulin receptor, trans-Golgi

  14. Unconventional secretion of tissue transglutaminase involves phospholipid-dependent delivery into recycling endosomes.

    Directory of Open Access Journals (Sweden)

    Evgeny A Zemskov

    2011-04-01

    Full Text Available Although endosomal compartments have been suggested to play a role in unconventional protein secretion, there is scarce experimental evidence for such involvement. Here we report that recycling endosomes are essential for externalization of cytoplasmic secretory protein tissue transglutaminase (tTG. The de novo synthesized cytoplasmic tTG does not follow the classical ER/Golgi-dependent secretion pathway, but is targeted to perinuclear recycling endosomes, and is delivered inside these vesicles prior to externalization. On its route to the cell surface tTG interacts with internalized β1 integrins inside the recycling endosomes and is secreted as a complex with recycled β1 integrins. Inactivation of recycling endosomes, blocking endosome fusion with the plasma membrane, or downregulation of Rab11 GTPase that controls outbound trafficking of perinuclear recycling endosomes, all abrogate tTG secretion. The initial recruitment of cytoplasmic tTG to recycling endosomes and subsequent externalization depend on its binding to phosphoinositides on endosomal membranes. These findings begin to unravel the unconventional mechanism of tTG secretion which utilizes the long loop of endosomal recycling pathway and indicate involvement of endosomal trafficking in non-classical protein secretion.

  15. Arf6, Rab11 and transferrin receptor define distinct populations of recycling endosomes.

    Science.gov (United States)

    Kobayashi, Hotaka; Fukuda, Mitsunori

    2013-09-01

    Recycling endosomes are key platforms for endocytic recycling that return internalized molecules back to the plasma membrane. To determine how recycling endosomes perform their functions, searching for proteins and lipids that specifically localized at recycling endosomes has often been performed by colocalization analyses between candidate molecules and conventional recycling endosome markers. However, it remains unclear whether all the conventional markers have identical localizations. Here we report finding that three well-known recycling endosome markers, i.e., Arf6, Rab11 and transferrin receptor (TfR), have different intracellular localizations in PC12 cells. The results of immunofluorescence analyses showed that the signals of endogenous Arf6, Rab11 and TfR in nerve growth factor-stimulated PC12 cells generally differed, although there was some overlapping. Our findings provide new information about recycling endosome markers, and they highlight the heterogeneity of recycling endosomes.

  16. Promyelocytic leukemia bodies tether to early endosomes during mitosis.

    Science.gov (United States)

    Palibrk, Vuk; Lång, Emma; Lång, Anna; Schink, Kay Oliver; Rowe, Alexander D; Bøe, Stig Ove

    2014-01-01

    During mitosis the nuclear envelope breaks down, leading to potential interactions between cytoplasmic and nuclear components. PML bodies are nuclear structures with tumor suppressor and antiviral functions. Early endosomes, on the other hand, are cytoplasmic vesicles involved in transport and growth factor signaling. Here we demonstrate that PML bodies form stable interactions with early endosomes immediately following entry into mitosis. The 2 compartments remain stably associated throughout mitosis and dissociate in the cytoplasm of newly divided daughter cells. We also show that a minor subset of PML bodies becomes anchored to the mitotic spindle poles during cell division. The study demonstrates a stable mitosis-specific interaction between a cytoplasmic and a nuclear compartment.

  17. Selective T-cell Ablation with Bismuth-213 Labeled Anti-TCR Alpha Beta as Nonmyeloablative Conditioning for Allogeneic Canine Marrow Transplantion

    International Nuclear Information System (INIS)

    Bethge, W. A.; Wilbur, D. Scott; Storb, R.; Hamlin, Donald K.; Santos, E. B.; Brechbiel, M. W.; Fisher, Darrell R.; Sandmaier, B. M.

    2003-01-01

    Two major immunological barriers, the host versus graft (HVG) and the graft versus host (GVH) reaction, must be overcome for successful allogeneic hematopoietic stem cell transplantation. T-cells are involved in these barriers in the major histocompatibility complex-identical settings. We hypothesized that selective ablation of T-cells using radioimmunotherapy, together with postgrafting immunosuppression, would ensure stable allogeneic engraftment. We developed a canine model of nonmyeloablative marrow transplantation in which host immune reactions are impaired by a single dose of 2 Gy total body irradiation (TBI), and where both GVH and residual HVG reactions are controlled by postgrafting immunosuppression with mycophenolate mofetil (MMF) and cyclosporine (CSP). We substituted the alpha-emitter bismuth-213 linked to a monoclonal antibody against TCR(alpha,beta)using the metal-binding chelate CHX-A-DTPA, for 2 Gy TBI. Biodistribution studies using a gamma-emitting indium-111-labeled anti-TCR mAb showed uptake primarily in blood, marrow, lymph nodes, spleen and liver. In a dosimetry study, 4 dogs were treated with 0.13-0.46 mg/kg TCR mAb labeled with 3.7-5.6 mCi/kg (137-207 MBq/kg) Bi-213. The treatment was administered in 6 injections on days -3 and -2 followed by transplantion of dog leukocyte antigen-identical marrow on day 0 and postgrafting immunosuppression with MMF and CSP. Therapy was well tolerated except for elevations of transaminases, which were transient in all but one dog. No other organ toxicities or signs of graft-versus-host-disease were noted. The dogs had prompt allogeneic hematopoietic engraftment and achieved stable mixed donor-host hematopoietic chimerism with donor contributions ranging from 5-55 % with >30 weeks follow up

  18. Selective T-cell Ablation with Bismuth-213 Labeled Anti-TCR Alpha Beta as Nonmyeloablative Conditionaing for Allogeneic Canine Marrow Transplantion

    Energy Technology Data Exchange (ETDEWEB)

    Bethge, W. A.; Wilbur, D. Scott; Storb, R.; Hamlin, Donald K.; Santos, E. B.; Brechbiel, M. W.; Fisher, Darrell R.; Sandmaier, B. M.

    2003-06-15

    Two major immunological barriers, the host versus graft (HVG) and the graft versus host (GVH) reaction, must be overcome for successful allogeneic hematopoietic stem cell transplantation. T-cells are involved in these barriers in the major histocompatibility complex-identical settings. We hypothesized that selective ablation of T-cells using radioimmunotherapy, together with postgrafting immunosuppression, would ensure stable allogeneic engraftment. We developed a canine model of nonmyeloablative marrow transplantation in which host immune reactions are impaired by a single dose of 2 Gy total body irradiation (TBI), and where both GVH and residual HVG reactions are controlled by postgrafting immunosuppression with mycophenolate mofetil (MMF) and cyclosporine (CSP). We substituted the alpha-emitter bismuth-213 linked to a monoclonal antibody against TCR(alpha,beta)using the metal-binding chelate CHX-A”-DTPA, for 2 Gy TBI. Biodistribution studies using a gamma-emitting indium-111-labeled anti-TCR mAb showed uptake primarily in blood, marrow, lymph nodes, spleen and liver. In a dosimetry study, 4 dogs were treated with 0.13-0.46 mg/kg TCR mAb labeled with 3.7-5.6 mCi/kg (137-207 MBq/kg) Bi-213. The treatment was administered in 6 injections on days -3 and -2 followed by transplantion of dog leukocyte antigen-identical marrow on day 0 and postgrafting immunosuppression with MMF and CSP. Therapy was well tolerated except for elevations of transaminases, which were transient in all but one dog. No other organ toxicities or signs of graft-versus-host-disease were noted. The dogs had prompt allogeneic hematopoietic engraftment and achieved stable mixed donor-host hematopoietic chimerism with donor contributions ranging from 5-55 % with >30 weeks follow up.

  19. Enhancing endosomal escape of transduced proteins by photochemical internalisation.

    Directory of Open Access Journals (Sweden)

    Kevin Mellert

    Full Text Available Induced internalisation of functional proteins into cultured cells has become an important aspect in a rising number of in vitro and in vivo assays. The endo-lysosomal entrapment of the transduced proteins remains the major problem in all transduction protocols. In this study we compared the efficiency, cytotoxicity and protein targeting of different commercially available transduction reagents by transducing a well-studied fluorescently labelled protein (Atto488-bovine serum albumin into cultured human sarcoma cells. The amount of internalised protein and toxicity differed between the different reagents, but the percentage of transduced cells was consistently high. Furthermore, in all protocols the signals of the transduced Atto488-BSA were predominantly punctual consistent with an endosomal localisation. To overcome the endosomal entrapment, the transduction protocols were combined with a photochemical internalisation (PCI treatment. Using this combination revealed that an endosomal disruption is highly effective in cell penetrating peptide (CPP mediated transduction, whereas lipid-mediated transductions lead to a lower signal spreading throughout the cytosol. No change in the signal distribution could be achieved in treatments using non-lipid polymers as a transduction reagent. Therefore, the combination of protein transduction protocols based on CPPs with the endosomolytic treatment PCI can facilitate protein transduction experiments in vitro.

  20. Enhancing endosomal escape of transduced proteins by photochemical internalisation.

    Science.gov (United States)

    Mellert, Kevin; Lamla, Markus; Scheffzek, Klaus; Wittig, Rainer; Kaufmann, Dieter

    2012-01-01

    Induced internalisation of functional proteins into cultured cells has become an important aspect in a rising number of in vitro and in vivo assays. The endo-lysosomal entrapment of the transduced proteins remains the major problem in all transduction protocols. In this study we compared the efficiency, cytotoxicity and protein targeting of different commercially available transduction reagents by transducing a well-studied fluorescently labelled protein (Atto488-bovine serum albumin) into cultured human sarcoma cells. The amount of internalised protein and toxicity differed between the different reagents, but the percentage of transduced cells was consistently high. Furthermore, in all protocols the signals of the transduced Atto488-BSA were predominantly punctual consistent with an endosomal localisation. To overcome the endosomal entrapment, the transduction protocols were combined with a photochemical internalisation (PCI) treatment. Using this combination revealed that an endosomal disruption is highly effective in cell penetrating peptide (CPP) mediated transduction, whereas lipid-mediated transductions lead to a lower signal spreading throughout the cytosol. No change in the signal distribution could be achieved in treatments using non-lipid polymers as a transduction reagent. Therefore, the combination of protein transduction protocols based on CPPs with the endosomolytic treatment PCI can facilitate protein transduction experiments in vitro.

  1. NEU3 Sialidase Protein Interactors in the Plasma Membrane and in the Endosomes*

    Science.gov (United States)

    Cirillo, Federica; Ghiroldi, Andrea; Fania, Chiara; Piccoli, Marco; Torretta, Enrica; Tettamanti, Guido; Gelfi, Cecilia; Anastasia, Luigi

    2016-01-01

    NEU3 sialidase has been shown to be a key player in many physio- and pathological processes, including cell differentiation, cellular response to hypoxic stress, and carcinogenesis. The enzyme, peculiarly localized on the outer leaflet of the plasma membrane, has been shown to be able to remove sialic acid residues from the gangliosides present on adjacent cells, thus creating cell to cell interactions. Nonetheless, herein we report that the enzyme localization is dynamically regulated between the plasma membrane and the endosomes, where a substantial amount of NEU3 is stored with low enzymatic activity. However, under opportune stimuli, NEU3 is shifted from the endosomes to the plasma membrane, where it greatly increases the sialidase activity. Finally, we found that NEU3 possesses also the ability to interact with specific proteins, many of which are different in each cell compartment. They were identified by mass spectrometry, and some selected ones were also confirmed by cross-immunoprecipitation with the enzyme, supporting NEU3 involvement in the cell stress response, protein folding, and intracellular trafficking. PMID:26987901

  2. Protein complexes and cholesterol in the control of late endosomal dynamicsCholesterol and multi-protein complexes in the control of late endosomal dynamics

    NARCIS (Netherlands)

    Kant, Rik Henricus Nicolaas van der

    2013-01-01

    Late endosomal transport is disrupted in several diseases such as Niemann-Pick type C, ARC syndrome and Alzheimer’s disease. This thesis describes the regulation of late endosomal dynamics by cholesterol and multi-protein complexes. We find that cholesterol acts as a cellular tomtom that steers the

  3. Human Papillomavirus 16 Infection Induces VAP-Dependent Endosomal Tubulation.

    Science.gov (United States)

    Siddiqa, Abida; Massimi, Paola; Pim, David; Broniarczyk, Justyna; Banks, Lawrence

    2018-03-15

    Human papillomavirus (HPV) infection involves complex interactions with the endocytic transport machinery, which ultimately facilitates the entry of the incoming viral genomes into the trans -Golgi network (TGN) and their subsequent nuclear entry during mitosis. The endosomal pathway is a highly dynamic intracellular transport system, which consists of vesicular compartments and tubular extensions, although it is currently unclear whether incoming viruses specifically alter the endocytic machinery. In this study, using MICAL-L1 as a marker for tubulating endosomes, we show that incoming HPV-16 virions induce a profound alteration in global levels of endocytic tubulation. In addition, we also show a critical requirement for the endoplasmic reticulum (ER)-anchored protein VAP in this process. VAP plays an essential role in actin nucleation and endosome-to-Golgi transport. Indeed, the loss of VAP results in a dramatic decrease in the level of endosomal tubulation induced by incoming HPV-16 virions. This is also accompanied by a marked reduction in virus infectivity. In VAP knockdown cells, we see that the defect in virus trafficking occurs after capsid disassembly but prior to localization at the trans -Golgi network, with the incoming virion-transduced DNA accumulating in Vps29/TGN46-positive hybrid vesicles. Taken together, these studies demonstrate that infection with HPV-16 virions induces marked alterations of endocytic transport pathways, some of which are VAP dependent and required for the endosome-to-Golgi transport of the incoming viral L2/DNA complex. IMPORTANCE Human papillomavirus infectious entry involves multiple interactions with the endocytic transport machinery. In this study, we show that incoming HPV-16 virions induce a dramatic increase in endocytic tubulation. This tubulation requires ER-associated VAP, which plays a critical role in ensuring the delivery of cargoes from the endocytic compartments to the trans -Golgi network. Indeed, the loss of

  4. Perioral Rejuvenation With Ablative Erbium Resurfacing.

    Science.gov (United States)

    Cohen, Joel L

    2015-11-01

    Since the introduction of the scanning full-field erbium laser, misconceptions regarding ablative erbium resurfacing have resulted in its being largely overshadowed by ablative fractional resurfacing. This case report illustrates the appropriateness of full-field erbium ablation for perioral resurfacing. A patient with profoundly severe perioral photodamage etched-in lines underwent full-field ablative perioral resurfacing with an erbium laser (Contour TRL, Sciton Inc., Palo Alto, CA) that allows separate control of ablation and coagulation. The pre-procedure consultations included evaluation of the severity of etched-in lines, and discussion of patient goals, expectations, and appropriate treatment options, as well as a review of patient photos and post-treatment care required. The author generally avoids full-field erbium ablation in patients with Fitzpatrick type IV and above. For each of 2 treatment sessions (separated by approximately 4 months), the patient received (12 cc plain 2% lidodaine) sulcus blocks before undergoing 4 passes with the erbium laser at 150 μ ablation, no coagulation, and then some very focal 30 μ ablation to areas of residual lines still visualized through the pinpoint bleeding. Similarly, full-field ablative resurfacing can be very reliable for significant wrinkles and creping in the lower eyelid skin--where often a single treatment of 80 μ ablation, 50 μ coagulation can lead to a nice improvement. Standardized digital imaging revealed significant improvement in deeply etched rhytides without significant adverse events. For appropriately selected patients requiring perioral (or periorbital) rejuvenation, full-field ablative erbium resurfacing is safe, efficacious and merits consideration.

  5. Dosimetric selection for helical tomotherapy based stereotactic ablative radiotherapy for early-stage non-small cell lung cancer or lung metastases.

    Directory of Open Access Journals (Sweden)

    Alexander Chi

    Full Text Available BACKGROUND: No selection criteria for helical tomotherapy (HT based stereotactic ablative radiotherapy (SABR to treat early stage non-small cell lung cancer (NSCLC or solitary lung metastases has been established. In this study, we investigate the dosimetric selection criteria for HT based SABR delivering 70 Gy in 10 fractions to avoid severe toxicity in the treatment of centrally located lesions when adequate target dose coverage is desired. MATERIALS AND METHODS: 78 HT-SABR plans for solitary lung lesions were created to prescribe 70 Gy in 10 fractions to the planning target volume (PTV. The PTV was set to have ≥95% PTV receiving 70 Gy in each case. The cases for which dose constraints for ≥1 OAR could not be met without compromising the target dose coverage were compared with cases for which all target and OAR dose constraints were met. RESULTS: There were 23 central lesions for which OAR dose constraints could not be met without compromising PTV dose coverage. Comparing to cases for which optimal HT-based SABR plans were generated, they were associated with larger tumor size (5.72±1.96 cm vs. 3.74±1.49 cm, p<0.0001, higher lung dose, increased number of immediately adjacent OARs ( 3.45±1.34 vs. 1.66±0.81, p<0.0001, and shorter distance to the closest OARs (GTV: 0.26±0.22 cm vs. 0.88±0.54 cm, p<0.0001; PTV 0.19±0.18 cm vs. 0.48±0.36 cm, p = 0.0001. CONCLUSION: Delivery of 70 Gy in 10 fractions with HT to meet all the given OAR and PTV dose constraints are most likely when the following parameters are met: lung lesions ≤3.78 cm (11.98 cc, ≤2 immediately adjacent OARs which are ≥0.45 cm from the gross lesion and ≥0.21 cm from the PTV.

  6. Rab9-dependent retrograde transport and endosomal sorting of the endopeptidase furin

    Science.gov (United States)

    Chia, Pei Zhi Cheryl; Gasnereau, Isabelle; Lieu, Zi Zhao; Gleeson, Paul A.

    2011-01-01

    The endopeptidase furin and the trans-Golgi network protein TGN38 are membrane proteins that recycle between the TGN and plasma membrane. TGN38 is transported by a retromer-dependent pathway from early endosomes to the TGN, whereas the intracellular transport of furin is poorly defined. Here we have identified the itinerary and transport requirements of furin. Using internalisation assays, we show that furin transits the early and late endosomes en route to the TGN. The GTPase Rab9 and the TGN golgin GCC185, components of the late endosome-to-TGN pathway, were required for efficient TGN retrieval of furin. By contrast, TGN38 trafficking was independent of Rab9 and GCC185. To identify the sorting signals for the early endosome-to-TGN pathway, the trafficking of furin–TGN38 chimeras was investigated. The diversion of furin from the Rab9-dependent late-endosome-to-TGN pathway to the retromer-dependent early-endosome-to-TGN pathway required both the transmembrane domain and cytoplasmic tail of TGN38. We present evidence to suggest that the length of the transmembrane domain is a contributing factor in endosomal sorting. Overall, these data show that furin uses the Rab9-dependent pathway from late endosomes and that retrograde transport directly from early endosomes is dependent on both the transmembrane domain and the cytoplasmic tail. PMID:21693586

  7. Effect of diphtheria toxin T-domain on endosomal pH

    Directory of Open Access Journals (Sweden)

    A. J. Labyntsev

    2015-08-01

    Full Text Available A key step in the mode of cytotoxic action of diphtheria toxin (DT is the transfer of its catalytic domain (Cd from endosomes into the cytosol. The main activity in this process is performed by the transport domain (Td, but the molecular mechanism of its action remains unknown. We have previously shown that Td can have some influence on the endosomal transport of DT. The aim of this work was to study the effect of diphtheria toxin on the toxin compartmentalization in the intracellular transporting pathway and endosomal pH. We used recombinant fragments of DT, which differed only by the presence of Td in their structure, fused with fluorescent proteins. It was shown that the toxin fragment with Td moved slower by the pathway early-late endosomes-lysosomes, and had a slightly different pattern of colocalization with endosomal markers than DT fragment without Td. In addition, endosomes containing DT fragments with Td had a constant pH of about 6.5 from the 10th to 50th minute of observation, for the same time endosomes containing DT fragments without Td demons­trated a decrease in pH from 6.3 to 5.5. These results indicate that Td inhibits acidification of endosomal medium. One of possible explanations for this may be the effect of the ion channel formed by the T-domain on the process of the endosomal acidification. This property of Td may not only inhibit maturation of endosomes but also inhibit activation of endosomal pH-dependent proteases, and this promotes successful transport of Cd into the cell cytosol.

  8. Proteolytic processing of epidermal growth factor within endosomes

    International Nuclear Information System (INIS)

    Gorman, R.M.; Savage, C.R. Jr.; Poretz, R.D.; Schaudies, R.P.

    1986-01-01

    The authors have reported previously that EGF enters 3 biochemically distinct non-lysosomal intracellular compartments prior to detection within lysosomes. Earlier studies have demonstrated that EGF is processes by sequential removal of 1, 4 and 1 aminoacyl residues at the C-terminus. The final form, which lacks the 6 residues, accumulates in secondary lysosomes. After subcellular fractionation of fibroblasts exposed to 125 I-EGF, ligand is detected with 3 non-lysosomal endocytic compartments and is fully processed prior to entrance into secondary lysosome. Following internalization, EGF enters an early endosomal compartment (E 1 ). The composition of the ligand (60%, -1 form; 40%, native form) represents an enhancement of the -1 form relative to that on the plasma membrane following the 90 min, 0 0 binding period. The proportion of different EGF forms in E 1 remains constant through the 2 min pulse and chase periods up to 30 min. However, in the ultimate endosomal compartment, E 4 , the proportion of the -6 form increases from 25% at 15 min to greater than 75% in 30 min, with a concomitant decrease of the -1 and -5 forms. Secondary lysosomes contain an EGF composition similar to that found in E 4 at 30 min. Accordingly, it appears that EGF is processed to the -6 form following passage through E 1 and during its tenure in E 4

  9. Mouse polyomavirus enters early endosomes, requires their acidic pH for productive infection, and meets transferrin cargo in rab11-positive endosomes

    Czech Academy of Sciences Publication Activity Database

    Liebl, D.; Difato, F.; Horníková, L.; Mannová, P.; Štokrová, Jitka; Forstová, J.

    2006-01-01

    Roč. 80, č. 9 (2006), s. 4610-4622 ISSN 0022-538X R&D Projects: GA ČR(CZ) GA204/03/0593; GA MŠk(CZ) LC545 Institutional research plan: CEZ:AV0Z50520514 Keywords : Polyomavirus internalization and trafficking * Early endosomes * Dependence of infection on endosomal pH Subject RIV: EB - Genetics ; Molecular Biology Impact factor: 5.341, year: 2006

  10. deep-orange and carnation define distinct stages in late endosomal biogenesis in Drosophila melanogaster.

    Science.gov (United States)

    Sriram, V; Krishnan, K S; Mayor, Satyajit

    2003-05-12

    Endosomal degradation is severely impaired in primary hemocytes from larvae of eye color mutants of Drosophila. Using high resolution imaging and immunofluorescence microscopy in these cells, products of eye color genes, deep-orange (dor) and carnation (car), are localized to large multivesicular Rab7-positive late endosomes containing Golgi-derived enzymes. These structures mature into small sized Dor-negative, Car-positive structures, which subsequently fuse to form tubular lysosomes. Defective endosomal degradation in mutant alleles of dor results from a failure of Golgi-derived vesicles to fuse with morphologically arrested Rab7-positive large sized endosomes, which are, however, normally acidified and mature with wild-type kinetics. This locates the site of Dor function to fusion of Golgi-derived vesicles with the large Rab7-positive endocytic compartments. In contrast, endosomal degradation is not considerably affected in car1 mutant; fusion of Golgi-derived vesicles and maturation of large sized endosomes is normal. However, removal of Dor from small sized Car-positive endosomes is slowed, and subsequent fusion with tubular lysosomes is abolished. Overexpression of Dor in car1 mutant aggravates this defect, implicating Car in the removal of Dor from endosomes. This suggests that, in addition to an independent role in fusion with tubular lysosomes, the Sec1p homologue, Car, regulates Dor function.

  11. Pulmonary ablation: a primer.

    Science.gov (United States)

    Roberton, Benjamin J; Liu, David; Power, Mark; Wan, John M C; Stuart, Sam; Klass, Darren; Yee, John

    2014-05-01

    Percutaneous image-guided thermal ablation is safe and efficacious in achieving local control and improving outcome in the treatment of both early stage non-small-cell lung cancer and pulmonary metastatic disease, in which surgical treatment is precluded by comorbidity, poor cardiorespiratory reserve, or unfavorable disease distribution. Radiofrequency ablation is the most established technology, but new thermal ablation technologies such as microwave ablation and cryoablation may offer some advantages. The use of advanced techniques, such as induced pneumothorax and the popsicle stick technique, or combining thermal ablation with radiotherapy, widens the treatment options available to the multidisciplinary team. The intent of this article is to provide the reader with a practical knowledge base of pulmonary ablation by concentrating on indications, techniques, and follow-up. Copyright © 2014 Canadian Association of Radiologists. Published by Elsevier Inc. All rights reserved.

  12. An Essential Role of Hrs/Vps27 in Endosomal Cholesterol Trafficking

    Directory of Open Access Journals (Sweden)

    Ximing Du

    2012-01-01

    Full Text Available The endosomal sorting complex required for transport (ESCRT plays a crucial role in the degradation of ubiquitinated endosomal membrane proteins. Here, we report that Hrs, a key protein of the ESCRT-0 complex, is required for the transport of low-density lipoprotein-derived cholesterol from endosomes to the endoplasmic reticulum. This function of Hrs in cholesterol transport is distinct from its previously defined role in lysosomal sorting and downregulation of membrane receptors via the ESCRT pathway. In line with this, knocking down other ESCRT proteins does not cause prominent endosomal cholesterol accumulation. Importantly, the localization and biochemical properties of key cholesterol-sorting proteins, NPC1 and NPC2, appear to be unchanged upon Hrs knockdown. Our data identify Hrs as a regulator of endosomal cholesterol trafficking and provide additional insights into the budding of intralumenal vesicles.

  13. Retrogradely Transported TrkA Endosomes Signal Locally within Dendrites to Maintain Sympathetic Neuron Synapses

    Directory of Open Access Journals (Sweden)

    Kathryn M. Lehigh

    2017-04-01

    Full Text Available Sympathetic neurons require NGF from their target fields for survival, axonal target innervation, dendritic growth and formation, and maintenance of synaptic inputs from preganglionic neurons. Target-derived NGF signals are propagated retrogradely, from distal axons to somata of sympathetic neurons via TrkA signaling endosomes. We report that a subset of TrkA endosomes that are transported from distal axons to cell bodies translocate into dendrites, where they are signaling competent and move bidirectionally, in close proximity to synaptic protein clusters. Using a strategy for spatially confined inhibition of TrkA kinase activity, we found that distal-axon-derived TrkA signaling endosomes are necessary within sympathetic neuron dendrites for maintenance of synapses. Thus, TrkA signaling endosomes have unique functions in different cellular compartments. Moreover, target-derived NGF mediates circuit formation and synapse maintenance through TrkA endosome signaling within dendrites to promote aggregation of postsynaptic protein complexes.

  14. Charcot-Marie-Tooth disease-linked protein SIMPLE functions with the ESCRT machinery in endosomal trafficking

    OpenAIRE

    Lee, Samuel M.; Chin, Lih-Shen; Li, Lian

    2012-01-01

    Mutations in small integral membrane protein of lysosome/late endosome (SIMPLE) cause autosomal dominant, Charcot-Marie-Tooth disease (CMT) type 1C. The cellular function of SIMPLE is unknown and the pathogenic mechanism of SIMPLE mutations remains elusive. Here, we report that SIMPLE interacted and colocalized with endosomal sorting complex required for transport (ESCRT) components STAM1, Hrs, and TSG101 on early endosomes and functioned with the ESCRT machinery in the control of endosome-to...

  15. PTP1B targets the endosomal sorting machinery

    DEFF Research Database (Denmark)

    Stuible, Matthew; Abella, Jasmine V; Feldhammer, Matthew

    2010-01-01

    Dephosphorylation and endocytic down-regulation are distinct processes that together control the signaling output of a variety of receptor tyrosine kinases (RTKs). PTP1B can directly dephosphorylate several RTKs, but it can also promote activation of downstream pathways through largely unknown...... mechanisms. These positive signaling functions likely contribute to the tumor-promoting effect of PTP1B in mouse cancer models. Here, we have identified STAM2, an endosomal protein involved in sorting activated RTKs for lysosomal degradation, as a substrate of PTP1B. PTP1B interacts with STAM2 at defined...... phosphotyrosine sites, and knockdown of PTP1B expression augments STAM2 phosphorylation. Intriguingly, manipulating the expression and phosphorylation state of STAM2 did not have a general effect on epidermal growth factor (EGF)-induced EGF receptor trafficking, degradation, or signaling. Instead, phosphorylated...

  16. Laser ablation principles and applications

    CERN Document Server

    1994-01-01

    Laser Ablation provides a broad picture of the current understanding of laser ablation and its many applications, from the views of key contributors to the field. Discussed are in detail the electronic processes in laser ablation of semiconductors and insulators, the post-ionization of laser-desorbed biomolecules, Fourier-transform mass spectroscopy, the interaction of laser radiation with organic polymers, laser ablation and optical surface damage, laser desorption/ablation with laser detection, and laser ablation of superconducting thin films.

  17. Stochastic acidification, activation of hemagglutinin and escape of influenza viruses from an endosome

    Science.gov (United States)

    Lagache, Thibault; Sieben, Christian; Meyer, Tim; Herrmann, Andreas; Holcman, David

    2017-06-01

    Influenza viruses enter the cell inside an endosome. During the endosomal journey, acidification triggers a conformational change of the virus spike protein hemagglutinin (HA) that results in escape of the viral genome from the endosome into the cytoplasm. It is still unclear how the interplay between acidification and HA conformation changes affects the kinetics of the viral endosomal escape. We develop here a stochastic model to estimate the change of conformation of HAs inside the endosome nanodomain. Using a Markov process, we model the arrival of protons to HA binding sites and compute the kinetics of their accumulation. We compute the Mean First Passage Time (MFPT) of the number of HA bound sites to a threshold, which is used to estimate the HA activation rate for a given pH concentration. The present analysis reveals that HA proton binding sites possess a high chemical barrier, ensuring a stability of the spike protein at sub-acidic pH. We predict that activating more than 3 adjacent HAs is necessary to trigger endosomal fusion and this configuration prevents premature release of viruses from early endosomes

  18. ER network homeostasis is critical for plant endosome streaming and endocytosis

    Science.gov (United States)

    Stefano, Giovanni; Renna, Luciana; Lai, YaShiuan; Slabaugh, Erin; Mannino, Nicole; Buono, Rafael A; Otegui, Marisa S; Brandizzi, Federica

    2015-01-01

    Eukaryotic cells internalize cargo at the plasma membrane via endocytosis, a vital process that is accomplished through a complex network of endosomal organelles. In mammalian cells, the ER is in close association with endosomes and regulates their fission. Nonetheless, the physiological role of such interaction on endocytosis is yet unexplored. Here, we probed the existence of ER–endosome association in plant cells and assayed its physiological role in endocytosis. Through live-cell imaging and electron microscopy studies, we established that endosomes are extensively associated with the plant ER, supporting conservation of interaction between heterotypic organelles in evolutionarily distant kingdoms. Furthermore, by analyzing ER–endosome dynamics in genetic backgrounds with defects in ER structure and movement, we also established that the ER network integrity is necessary for homeostasis of the distribution and streaming of various endosome populations as well as for efficient endocytosis. These results support a novel model that endocytosis homeostasis depends on a spatiotemporal control of the endosome dynamics dictated by the ER membrane network. PMID:27462431

  19. Genetic reconstitution of the human Adenovirus type 2 temperature-sensitive 1 mutant defective in endosomal escape

    Directory of Open Access Journals (Sweden)

    Gastaldelli Michele

    2009-10-01

    Full Text Available Abstract Human Adenoviruses infect the upper and lower respiratory tracts, the urinary and digestive tracts, lymphoid systems and heart, and give rise to epidemic conjunctivitis. More than 51 human serotypes have been identified to-date, and classified into 6 species A-F. The species C Adenoviruses Ad2 and Ad5 (Ad2/5 cause upper and lower respiratory disease, but how viral structure relates to the selection of particular infectious uptake pathways is not known. An adenovirus mutant, Ad2-ts1 had been isolated upon chemical mutagenesis in the past, and shown to have unprocessed capsid proteins. Ad2-ts1 fails to package the viral protease L3/p23, and Ad2-ts1 virions do not efficiently escape from endosomes. It had been suggested that the C22187T point mutation leading to the substitution of the conserved proline 137 to leucine (P137L in the L3/p23 protease was at least in part responsible for this phenotype. To clarify if the C22187T mutation is necessary and sufficient for the Ad2-ts1 phenotype, we sequenced the genes encoding the structural proteins of Ad2-ts1, and confirmed that the Ad2-ts1 DNA carries the point mutation C22187T. Introduction of C22187T to the wild-type Ad2 genome in a bacterial artificial chromosome (Ad2-BAC gave Ad2-BAC46 virions with the full Ad2-ts1 phenotype. Reversion of Ad2-BAC46 gave wild-type Ad2 particles indicating that P137L is necessary and sufficient for the Ad2-ts1 phenotype. The kinetics of Ad2-ts1 uptake into cells were comparable to Ad2 suggesting similar endocytic uptake mechanisms. Surprisingly, infectious Ad2 or Ad5 but not Ad2-ts1 uptake required CALM (clathrin assembly lymphoid myeloid protein, which controls clathrin-mediated endocytosis and membrane transport between endosomes and the trans-Golgi-network. The data show that no other mutations than P137L in the viral protease are necessary to give rise to particles that are defective in capsid processing and endosomal escape. This provides a basis for

  20. Radiofrequency ablation in dermatology

    Directory of Open Access Journals (Sweden)

    Sachdeva Silonie

    2007-01-01

    Full Text Available Radiofreqeuency ablation is a versatile dermatosurgical procedure used for surgical management of skin lesions by using various forms of alternating current at an ultra high frequency. The major modalities in radiofrequency are electrosection, electrocoagulation, electrodessication and fulguration. The use of radiofrequency ablation in dermatosurgical practice has gained importance in recent years as it can be used to treat most of the skin lesions with ease in less time with clean surgical field due to adequate hemostasis and with minimal side effects and complications. This article focuses on the major tissue effects and factors influencing radiofrequency ablation and its application for various dermatological conditions.

  1. Recycling endosomes in apical plasma membrane domain formation and epithelial cell polarity

    NARCIS (Netherlands)

    Golachowska, Magdalena R.; Hoekstra, Dick; van IJzendoorn, Sven C. D.

    2010-01-01

    Recycling endosomes have taken central stage in the intracellular sorting and polarized trafficking of apical and basolateral plasma membrane components. Molecular players in the underlying mechanisms are now emerging, including small GTPases, class V myosins and adaptor proteins. In particular,

  2. An ER-Associated Pathway Defines Endosomal Architecture for Controlled Cargo Transport

    NARCIS (Netherlands)

    Jongsma, Marlieke L. M.; Berlin, Ilana; Wijdeven, Ruud H. M.; Janssen, Lennert; Janssen, George M. C.; Garstka, Malgorzata A.; Janssen, Hans; Mensink, Mark; van Veelen, Peter A.; Spaapen, Robbert M.; Neefjes, Jacques

    2016-01-01

    Through a network of progressively maturing vesicles, the endosomal system connects the cell's interior with extracellular space. Intriguingly, this network exhibits a bilateral architecture, comprised of a relatively immobile perinuclear vesicle "cloud" and a highly dynamic peripheral contingent.

  3. Erythroid cell mitochondria receive endosomal iron by a "kiss-and-run" mechanism.

    Science.gov (United States)

    Hamdi, Amel; Roshan, Tariq M; Kahawita, Tanya M; Mason, Anne B; Sheftel, Alex D; Ponka, Prem

    2016-12-01

    In erythroid cells, more than 90% of transferrin-derived iron enters mitochondria where ferrochelatase inserts Fe 2+ into protoporphyrin IX. However, the path of iron from endosomes to mitochondrial ferrochelatase remains elusive. The prevailing opinion is that, after its export from endosomes, the redox-active metal spreads into the cytosol and mysteriously finds its way into mitochondria through passive diffusion. In contrast, this study supports the hypothesis that the highly efficient transport of iron toward ferrochelatase in erythroid cells requires a direct interaction between transferrin-endosomes and mitochondria (the "kiss-and-run" hypothesis). Using a novel method (flow sub-cytometry), we analyze lysates of reticulocytes after labeling these organelles with different fluorophores. We have identified a double-labeled population definitively representing endosomes interacting with mitochondria, as demonstrated by confocal microscopy. Moreover, we conclude that this endosome-mitochondrion association is reversible, since a "chase" with unlabeled holotransferrin causes a time-dependent decrease in the size of the double-labeled population. Importantly, the dissociation of endosomes from mitochondria does not occur in the absence of holotransferrin. Additionally, mutated recombinant holotransferrin, that cannot release iron, significantly decreases the uptake of 59 Fe by reticulocytes and diminishes 59 Fe incorporation into heme. This suggests that endosomes, which are unable to provide iron to mitochondria, cause a "traffic jam" leading to decreased endocytosis of holotransferrin. Altogether, our results suggest that a molecular mechanism exists to coordinate the iron status of endosomal transferrin with its trafficking. Besides its contribution to the field of iron metabolism, this study provides evidence for a new intracellular trafficking pathway of organelles. Copyright © 2016 Elsevier B.V. All rights reserved.

  4. deep-orange and carnation define distinct stages in late endosomal biogenesis in Drosophila melanogaster

    OpenAIRE

    Sriram, V.; Krishnan, K.S.; Mayor, Satyajit

    2003-01-01

    Endosomal degradation is severely impaired in primary hemocytes from larvae of eye color mutants of Drosophila. Using high resolution imaging and immunofluorescence microscopy in these cells, products of eye color genes, deep-orange (dor) and carnation (car), are localized to large multivesicular Rab7-positive late endosomes containing Golgi-derived enzymes. These structures mature into small sized Dor-negative, Car-positive structures, which subsequently fuse to form tubular lysosomes. Defec...

  5. Antigen processing and remodeling of the endosomal pathway: requirements for antigen cross-presentation.

    Science.gov (United States)

    Compeer, Ewoud Bernardus; Flinsenberg, Thijs Willem Hendrik; van der Grein, Susanna Geertje; Boes, Marianne

    2012-01-01

    Cross-presentation of endocytosed antigen as peptide/class I major histocompatibility complex complexes plays a central role in the elicitation of CD8(+) T cell clones that mediate anti-viral and anti-tumor immune responses. While it has been clear that there are specific subsets of professional antigen presenting cells capable of antigen cross-presentation, identification of mechanisms involved is still ongoing. Especially amongst dendritic cells (DC), there are specialized subsets that are highly proficient at antigen cross-presentation. We here present a focused survey on the cell biological processes in the endosomal pathway that support antigen cross-presentation. This review highlights DC-intrinsic mechanisms that facilitate the cross-presentation of endocytosed antigen, including receptor-mediated uptake, maturation-induced endosomal sorting of membrane proteins, dynamic remodeling of endosomal structures and cell surface-directed endosomal trafficking. We will conclude with the description of pathogen-induced deviation of endosomal processing, and discuss how immune evasion strategies pertaining endosomal trafficking may preclude antigen cross-presentation.

  6. A Role for EHD4 in the Regulation of Early Endosomal Transport

    Science.gov (United States)

    Sharma, Mahak; Naslavsky, Naava; Caplan, Steve

    2009-01-01

    All four of the C-terminal Eps15 homology domain (EHD) proteins have been implicated in the regulation of endocytic trafficking. However, the high level of amino acid sequence identity among these proteins has made it challenging to elucidate the precise function of individual EHD proteins. We demonstrate here with specific peptide antibodies that endogenous EHD4 localizes to Rab5-, early embryonic antigen 1 (EEA1)- and Arf6-containing endosomes and colocalizes with internalized transferrin in the cell periphery. Knock-down of EHD4 expression by both small interfering RNA and short hairpin RNA leads to the generation of enlarged early endosomal structures that contain Rab5 and EEA1 as well as internalized transferrin or major histocompatibility complex class I molecules. In addition, cargo destined for degradation, such as internalized low-density lipoprotein, also accumulates in the enlarged early endosomes in EHD4-depleted cells. Moreover, we have demonstrated that these enlarged early endosomes are enriched in levels of the activated GTP-bound Rab5. Finally, we show that endogenous EHD4 and EHD1 interact in cells, suggesting coordinated involvement in the regulation of receptor transport along the early endosome to endocytic recycling compartment axis. The results presented herein provide evidence that EHD4 is involved in the control of trafficking at the early endosome and regulates exit of cargo toward both the recycling compartment and the late endocytic pathway. PMID:18331452

  7. Enhanced Endosomal Escape by Light-Fueled Liquid-Metal Transformer.

    Science.gov (United States)

    Lu, Yue; Lin, Yiliang; Chen, Zhaowei; Hu, Quanyin; Liu, Yang; Yu, Shuangjiang; Gao, Wei; Dickey, Michael D; Gu, Zhen

    2017-04-12

    Effective endosomal escape remains as the "holy grail" for endocytosis-based intracellular drug delivery. To date, most of the endosomal escape strategies rely on small molecules, cationic polymers, or pore-forming proteins, which are often limited by the systemic toxicity and lack of specificity. We describe here a light-fueled liquid-metal transformer for effective endosomal escape-facilitated cargo delivery via a chemical-mechanical process. The nanoscale transformer can be prepared by a simple approach of sonicating a low-toxicity liquid-metal. When coated with graphene quantum dots (GQDs), the resulting nanospheres demonstrate the ability to absorb and convert photoenergy to drive the simultaneous phase separation and morphological transformation of the inner liquid-metal core. The morphological transformation from nanospheres to hollow nanorods with a remarkable change of aspect ratio can physically disrupt the endosomal membrane to promote endosomal escape of payloads. This metal-based nanotransformer equipped with GQDs provides a new strategy for facilitating effective endosomal escape to achieve spatiotemporally controlled drug delivery with enhanced efficacy.

  8. Overexpression of Rab22a hampers the transport between endosomes and the Golgi apparatus

    International Nuclear Information System (INIS)

    Mesa, Rosana; Magadan, Javier; Barbieri, Alejandro; Lopez, Cecilia; Stahl, Philip D.; Mayorga, Luis S.

    2005-01-01

    The transport and sorting of soluble and membrane-associated macromolecules arriving at endosomal compartments require a complex set of Rab proteins. Rab22a has been localized to the endocytic compartment; however, very little is known about the function of Rab22a and inconsistent results have been reported in studies performed in different cell lines. To characterize the function of Rab22a in endocytic transport, the wild-type protein (Rab22a WT), a hydrolysis-deficient mutant (Rab22a Q64L), and a mutant with reduced affinity for GTP (Rab22a S19N) were expressed in CHO cells. None of the three Rab22a constructs affected the transport of rhodamine-dextran to lysosomes, the digestion of internalized proteins, or the lysosomal localization of cathepsin D. In contrast with the mild effect of Rab22a on the endosome-lysosome route, cells expressing Rab22a WT and Rab22a Q64L presented a strong delay in the retrograde transport of cholera toxin from endosomes to the Golgi apparatus. Moreover, these cells accumulated the cation independent mannose 6-phosphate receptor in endosomes. These observations indicate that Rab22a can affect the trafficking from endosomes to the Golgi apparatus probably by promoting fusion among endosomes and impairing the proper segregation of membrane domains required for targeting to the trans-Golgi network (TGN)

  9. Post-Golgi anterograde transport requires GARP-dependent endosome-to-TGN retrograde transport

    Science.gov (United States)

    Hirata, Tetsuya; Fujita, Morihisa; Nakamura, Shota; Gotoh, Kazuyoshi; Motooka, Daisuke; Murakami, Yoshiko; Maeda, Yusuke; Kinoshita, Taroh

    2015-01-01

    The importance of endosome-to–trans-Golgi network (TGN) retrograde transport in the anterograde transport of proteins is unclear. In this study, genome-wide screening of the factors necessary for efficient anterograde protein transport in human haploid cells identified subunits of the Golgi-associated retrograde protein (GARP) complex, a tethering factor involved in endosome-to-TGN transport. Knockout (KO) of each of the four GARP subunits, VPS51–VPS54, in HEK293 cells caused severely defective anterograde transport of both glycosylphosphatidylinositol (GPI)-anchored and transmembrane proteins from the TGN. Overexpression of VAMP4, v-SNARE, in VPS54-KO cells partially restored not only endosome-to-TGN retrograde transport, but also anterograde transport of both GPI-anchored and transmembrane proteins. Further screening for genes whose overexpression normalized the VPS54-KO phenotype identified TMEM87A, encoding an uncharacterized Golgi-resident membrane protein. Overexpression of TMEM87A or its close homologue TMEM87B in VPS54-KO cells partially restored endosome-to-TGN retrograde transport and anterograde transport. Therefore GARP- and VAMP4-dependent endosome-to-TGN retrograde transport is required for recycling of molecules critical for efficient post-Golgi anterograde transport of cell-surface integral membrane proteins. In addition, TMEM87A and TMEM87B are involved in endosome-to-TGN retrograde transport. PMID:26157166

  10. PIKfyve mediates the motility of late endosomes and lysosomes in neuronal dendrites.

    Science.gov (United States)

    Tsuruta, Fuminori; Dolmetsch, Ricardo E

    2015-09-25

    The endosome/lysosome system in the nervous system is critically important for a variety of neuronal functions such as neurite outgrowth, retrograde transport, and synaptic plasticity. In neurons, the endosome/lysosome system is crucial for the activity-dependent internalization of membrane proteins and contributes to the regulation of lipid level on the plasma membrane. Although homeostasis of membrane dynamics plays important roles in the properties of central nervous systems, it has not been elucidated how endosome/lysosome system is regulated. Here, we report that phosphatidylinositol 3-phosphate 5-kinase (PIKfyve) mediates the motility of late endosomes and lysosomes in neuronal dendrites. Endosomes and lysosomes are highly motile in resting neurons, however knockdown of PIKfyve led to a significant reduction in late endosomes and lysosomes motility. We also found that vesicle acidification is crucial for their motility and PIKfyve is associated with this process indirectly. These data suggest that PIKfyve mediates vesicle motility through the regulation of vesicle integrity in neurons. Copyright © 2015 Elsevier Ireland Ltd. All rights reserved.

  11. Moldable cork ablation material

    Science.gov (United States)

    1977-01-01

    A successful thermal ablative material was manufactured. Moldable cork sheets were tested for density, tensile strength, tensile elongation, thermal conductivity, compression set, and specific heat. A moldable cork sheet, therefore, was established as a realistic product.

  12. The ESCRT regulator Did2 maintains the balance between long-distance endosomal transport and endocytic trafficking.

    Directory of Open Access Journals (Sweden)

    Carl Haag

    2017-04-01

    Full Text Available In highly polarised cells, like fungal hyphae, early endosomes function in both endocytosis as well as long-distance transport of various cargo including mRNA and protein complexes. However, knowledge on the crosstalk between these seemingly different trafficking processes is scarce. Here, we demonstrate that the ESCRT regulator Did2 coordinates endosomal transport in fungal hyphae of Ustilago maydis. Loss of Did2 results in defective vacuolar targeting, less processive long-distance transport and abnormal shuttling of early endosomes. Importantly, the late endosomal protein Rab7 and vacuolar protease Prc1 exhibit increased shuttling on these aberrant endosomes suggesting defects in endosomal maturation and identity. Consistently, molecular motors fail to attach efficiently explaining the disturbed processive movement. Furthermore, the endosomal mRNP linker protein Upa1 is hardly present on endosomes resulting in defects in long-distance mRNA transport. In conclusion, the ESCRT regulator Did2 coordinates precise maturation of endosomes and thus provides the correct membrane identity for efficient endosomal long-distance transport.

  13. Aggregation of endosomal-vacuolar compartments in the Aovps24-deleted strain in the filamentous fungus Aspergillus oryzae

    International Nuclear Information System (INIS)

    Tatsumi, Akinori; Shoji, Jun-ya; Kikuma, Takashi; Arioka, Manabu; Kitamoto, Katsuhiko

    2007-01-01

    Previously, we found that deletion of Aovps24, an ortholog of Saccharomyces cerevisiae VPS24, that encodes an ESCRT (endosomal sorting complex required for transport)-III component required for late endosomal function results in fragmented and aggregated vacuoles. Although defective late endosomal function is likely responsible for this phenotype, critical lack of our knowledge on late endosomes in filamentous fungi prevented us from further characterization. In this study, we identified late endosomes of Aspergillus oryzae, by expressing a series of fusion proteins of fluorescent proteins with orthologs of late endosomal proteins. Using these fusion proteins as markers, we observed late endosomes in the wild type strain and the Aovps24 disruptant and demonstrated that late endosomes are aberrantly aggregated in the Aovps24 disruptant. Moreover, we revealed that the aggregated late endosomes have features of vacuoles as well. As deletion of another ESCRT-III component-encoding gene, Aovps2, resulted in similar phenotypes to that in the Aovps24 disruptant, phenotypes of the Aovps24 disruptant are probably due to defective late endosomal function

  14. Comparison of the Three NIF Ablators

    Energy Technology Data Exchange (ETDEWEB)

    Kritcher, A. L. [Lawrence Livermore National Lab. (LLNL), Livermore, CA (United States); Clark, D. S. [Lawrence Livermore National Lab. (LLNL), Livermore, CA (United States); Haan, S. W. [Lawrence Livermore National Lab. (LLNL), Livermore, CA (United States); Yi, S. A. [Los Alamos National Lab. (LANL), Los Alamos, NM (United States); Zylstra, A. B. [Los Alamos National Lab. (LANL), Los Alamos, NM (United States); Ralph, J. E. [Lawrence Livermore National Lab. (LLNL), Livermore, CA (United States); Weber, C. R. [Lawrence Livermore National Lab. (LLNL), Livermore, CA (United States)

    2017-11-01

    Indirect drive implosion experiments on NIF have now been performed using three different ablator materials: glow discharge polymer (GDP) or CH, high density carbon (HDC, which we also refer to as diamond), and sputtered beryllium (Be). It has been appreciated for some time that each of these materials has specific advantages and disadvantages as an ICF ablator.[1-4] In light of experiments conducted on NIF in the last few years, how do these ablators compare? Given current understanding, is any ablator more or less likely to reach ignition on NIF? Has the understanding of their respective strengths and weaknesses changed since NIF experiments began? How are those strengths and weaknesses highlighted by implosion designs currently being tested or planned for testing soon? This document aims to address these questions by combining modern simulation results with a survey of the current experimental data base. More particularly, this document is meant to fulfill an L2 Milestone for FY17 to “Document our understanding of the relative advantages and disadvantages of CH, HDC, and Be designs.” Note that this document does not aim to recommend a down-selection of the current three ablator choices. It is intended only to gather and document the current understanding of the differences between these ablators and thereby inform the choices made in planning future implosion experiments. This document has two themes: (i) We report on a reanalysis project in which post-shot simulations were done on a common basis for layered shots using each ablator. This included data from keyholes, 2D ConA, and so forth, from each campaign, leading up to the layered shots. (“Keyholes” are shots dedicated to measuring the shock timing in a NIF target, as described in Ref. 5. “2DConAs” are backlit implosions in which the symmetry of the implosion is measured between about half and full convergence, as described in Ref. 6.) This set of common-basis postshot simulations is compared to

  15. Syndapin/SDPN-1 is required for endocytic recycling and endosomal actin association in the Caenorhabditis elegans intestine

    Science.gov (United States)

    Gleason, Adenrele M.; Nguyen, Ken C. Q.; Hall, David H.; Grant, Barth D.

    2016-01-01

    Syndapin/pascin-family F-BAR domain proteins bind directly to membrane lipids and are associated with actin dynamics at the plasma membrane. Previous reports also implicated mammalian syndapin 2 in endosome function during receptor recycling, but precise analysis of a putative recycling function for syndapin in mammalian systems is difficult because of its effects on the earlier step of endocytic uptake and potential redundancy among the three separate genes that encode mammalian syndapin isoforms. Here we analyze the endocytic transport function of the only Caenorhabditis elegans syndapin, SDPN-1. We find that SDPN-1 is a resident protein of the early and basolateral recycling endosomes in the C. elegans intestinal epithelium, and sdpn-1 deletion mutants display phenotypes indicating a block in basolateral recycling transport. sdpn-1 mutants accumulate abnormal endosomes positive for early endosome and recycling endosome markers that are normally separate, and such endosomes accumulate high levels of basolateral recycling cargo. Furthermore, we observed strong colocalization of endosomal SDPN-1 with the F-actin biosensor Lifeact and found that loss of SDPN-1 greatly reduced Lifeact accumulation on early endosomes. Taken together, our results provide strong evidence for an in vivo function of syndapin in endocytic recycling and suggest that syndapin promotes transport via endosomal fission. PMID:27630264

  16. Antigen processing and remodeling of the endosomal pathway: requirements for antigen cross-presentation.

    Directory of Open Access Journals (Sweden)

    Ewoud Bernardus Compeer

    2012-03-01

    Full Text Available The cross-presentation of endocytosed antigen as peptide/class I MHC complexes plays a central role in the elicitation of CD8+ T cell clones that mediate anti-viral and anti-tumor immune responses. While it has been clear that there are specific subsets of professional antigen presenting cells (APC capable of antigen cross-presentation, description of mechanisms involved is still ongoing. Especially amongst dendritic cells (DC, there are specialized subsets that are highly proficient at antigen cross-presentation. We here present a focused survey on the cell biological processes in the endosomal pathway that support antigen cross-presentation. This review highlight DC-intrinsic mechanisms that facilitate the cross-presentation of endocytosed antigen, including receptor-mediated uptake, recycling and maturation including the sorting of membrane proteins, dynamic remodeling of endosomal structures and cell-surface directed endosomal trafficking. We will conclude with description of pathogen-induced deviation of endosomal processing, and discuss how immune evasion strategies pertaining endosomal trafficking may preclude antigen cross-presentation.

  17. A FYVE zinc finger domain protein specifically links mRNA transport to endosome trafficking

    Science.gov (United States)

    Pohlmann, Thomas; Baumann, Sebastian; Haag, Carl; Albrecht, Mario; Feldbrügge, Michael

    2015-01-01

    An emerging theme in cellular logistics is the close connection between mRNA and membrane trafficking. A prominent example is the microtubule-dependent transport of mRNAs and associated ribosomes on endosomes. This coordinated process is crucial for correct septin filamentation and efficient growth of polarised cells, such as fungal hyphae. Despite detailed knowledge on the key RNA-binding protein and the molecular motors involved, it is unclear how mRNAs are connected to membranes during transport. Here, we identify a novel factor containing a FYVE zinc finger domain for interaction with endosomal lipids and a new PAM2-like domain required for interaction with the MLLE domain of the key RNA-binding protein. Consistently, loss of this FYVE domain protein leads to specific defects in mRNA, ribosome, and septin transport without affecting general functions of endosomes or their movement. Hence, this is the first endosomal component specific for mRNP trafficking uncovering a new mechanism to couple mRNPs to endosomes. DOI: http://dx.doi.org/10.7554/eLife.06041.001 PMID:25985087

  18. Vps33b pathogenic mutations preferentially affect VIPAS39/SPE-39-positive endosomes.

    Science.gov (United States)

    Tornieri, Karine; Zlatic, Stephanie A; Mullin, Ariana P; Werner, Erica; Harrison, Robert; L'hernault, Steven W; Faundez, Victor

    2013-12-20

    Mutations in Vps33 isoforms cause pigment dilution in mice (Vps33a, buff) and Drosophila (car) and the neurogenic arthrogryposis, renal dysfunction and cholestasis syndrome in humans (ARC1, VPS33B). The later disease is also caused by mutations in VIPAS39, (Vps33b interacting protein, apical-basolateral polarity regulator, SPE-39 homolog; ARC2), a protein that interacts with the HOmotypic fusion and Protein Sorting (HOPS) complex, a tether necessary for endosome-lysosome traffic. These syndromes offer insight into fundamental endosome traffic processes unique to metazoans. However, the molecular and cellular mechanisms underlying these mutant phenotypes remain poorly understood. Here we investigate interactions of wild-type and disease-causing mutations in VIPAS39/SPE-39 and Vps33b by yeast two hybrid, immunoprecipitation and quantitative fluorescent microscopy. We find that although few mutations prevent interaction between VIPAS39/SPE-39 and Vps33b, some mutants fragment VIPAS39/SPE-39-positive endosomes, but all mutants alter the subcellular localization of Vps33b to VIPAS39/SPE-39-positive endosomes. Our data suggest that the ARC syndrome may result through impaired VIPAS39/SPE-39 and Vps33b-dependent endosomal maturation or fusion.

  19. A FYVE zinc finger domain protein specifically links mRNA transport to endosome trafficking.

    Science.gov (United States)

    Pohlmann, Thomas; Baumann, Sebastian; Haag, Carl; Albrecht, Mario; Feldbrügge, Michael

    2015-05-18

    An emerging theme in cellular logistics is the close connection between mRNA and membrane trafficking. A prominent example is the microtubule-dependent transport of mRNAs and associated ribosomes on endosomes. This coordinated process is crucial for correct septin filamentation and efficient growth of polarised cells, such as fungal hyphae. Despite detailed knowledge on the key RNA-binding protein and the molecular motors involved, it is unclear how mRNAs are connected to membranes during transport. Here, we identify a novel factor containing a FYVE zinc finger domain for interaction with endosomal lipids and a new PAM2-like domain required for interaction with the MLLE domain of the key RNA-binding protein. Consistently, loss of this FYVE domain protein leads to specific defects in mRNA, ribosome, and septin transport without affecting general functions of endosomes or their movement. Hence, this is the first endosomal component specific for mRNP trafficking uncovering a new mechanism to couple mRNPs to endosomes.

  20. Femtosecond laser ablation profile near an interface: Analysis based on the correlation with superficial properties of individual materials

    Energy Technology Data Exchange (ETDEWEB)

    Nicolodelli, Gustavo, E-mail: nicolodelli@ursa.ifsc.usp.br [Instituto de Fisica de Sao Carlos, University of Sao Paulo, Grupo de Optica, Av. Trabalhador Sancarlense 400, P.O. Box 369, CEP 13560-970, Sao Carlos, SP (Brazil); Kurachi, Cristina; Bagnato, Vanderlei Salvador [Instituto de Fisica de Sao Carlos, University of Sao Paulo, Grupo de Optica, Av. Trabalhador Sancarlense 400, P.O. Box 369, CEP 13560-970, Sao Carlos, SP (Brazil)

    2011-01-15

    Femtosecond laser ablation of materials is turning to be an important tool for micromachining as well as for selective removal of biological tissues. In a great number of applications, laser ablation has to process through interfaces separating media of different properties. The investigation of the ablation behavior within materials and passing through interfaces is the main aim of this study. Especially, the analysis of the discontinuity in the ablation profile close to interfaces between distinct materials can reveal some of the phenomena involved in the formation of an ablated microcavity geometry. We have used a method that correlates the ablation cross sectional area with the local laser intensity. The effective intensity ablation properties were obtained from surface ablation data of distinct materials. The application of this method allows the prediction of the occurrence of a size discontinuity in the ablation geometry at the interface of distinct media, a fact which becomes important when planning applications in different media.

  1. Thermoresponsive pegylated bubble liposome nanovectors for efficient siRNA delivery via endosomal escape

    KAUST Repository

    Alamoudi, Kholod; Martins, Patricia; Croissant, Jonas G.; Patil, Sachin; Omar, Haneen; Khashab, Niveen M.

    2017-01-01

    Improving the delivery of siRNA into cancer cells via bubble liposomes. Designing a thermoresponsive pegylated liposome through the introduction of ammonium bicarbonate salt into liposomes so as to control their endosomal escape for gene therapy.A sub-200 nm nanovector was fully characterized and examined for cellular uptake, cytotoxicity, endosomal escape and gene silencing.The siRNA-liposomes were internalized into cancer cells within 5 min and then released siRNAs in the cytosol prior to lysosomal degradation upon external temperature elevation. This was confirmed by confocal bioimaging and gene silencing reaching up to 90% and further demonstrated by the protein inhibition of both target genes.The thermoresponsiveness of ammonium bicarbonate containing liposomes enabled the rapid endosomal escape of the particles and resulted in an efficient gene silencing.

  2. Cell polarity and patterning by PIN trafficking through early endosomal compartments in Arabidopsis thaliana.

    Directory of Open Access Journals (Sweden)

    Hirokazu Tanaka

    2013-05-01

    Full Text Available PIN-FORMED (PIN proteins localize asymmetrically at the plasma membrane and mediate intercellular polar transport of the plant hormone auxin that is crucial for a multitude of developmental processes in plants. PIN localization is under extensive control by environmental or developmental cues, but mechanisms regulating PIN localization are not fully understood. Here we show that early endosomal components ARF GEF BEN1 and newly identified Sec1/Munc18 family protein BEN2 are involved in distinct steps of early endosomal trafficking. BEN1 and BEN2 are collectively required for polar PIN localization, for their dynamic repolarization, and consequently for auxin activity gradient formation and auxin-related developmental processes including embryonic patterning, organogenesis, and vasculature venation patterning. These results show that early endosomal trafficking is crucial for cell polarity and auxin-dependent regulation of plant architecture.

  3. Thermoresponsive pegylated bubble liposome nanovectors for efficient siRNA delivery via endosomal escape

    KAUST Repository

    Alamoudi, Kholod

    2017-05-19

    Improving the delivery of siRNA into cancer cells via bubble liposomes. Designing a thermoresponsive pegylated liposome through the introduction of ammonium bicarbonate salt into liposomes so as to control their endosomal escape for gene therapy.A sub-200 nm nanovector was fully characterized and examined for cellular uptake, cytotoxicity, endosomal escape and gene silencing.The siRNA-liposomes were internalized into cancer cells within 5 min and then released siRNAs in the cytosol prior to lysosomal degradation upon external temperature elevation. This was confirmed by confocal bioimaging and gene silencing reaching up to 90% and further demonstrated by the protein inhibition of both target genes.The thermoresponsiveness of ammonium bicarbonate containing liposomes enabled the rapid endosomal escape of the particles and resulted in an efficient gene silencing.

  4. Laser Ablation for Cancer: Past, Present and Future

    Science.gov (United States)

    Schena, Emiliano; Saccomandi, Paola; Fong, Yuman

    2017-01-01

    Laser ablation (LA) is gaining acceptance for the treatment of tumors as an alternative to surgical resection. This paper reviews the use of lasers for ablative and surgical applications. Also reviewed are solutions aimed at improving LA outcomes: hyperthermal treatment planning tools and thermometric techniques during LA, used to guide the surgeon in the choice and adjustment of the optimal laser settings, and the potential use of nanoparticles to allow biologic selectivity of ablative treatments. Promising technical solutions and a better knowledge of laser-tissue interaction should allow LA to be used in a safe and effective manner as a cancer treatment. PMID:28613248

  5. Laser Ablation for Cancer: Past, Present and Future

    Directory of Open Access Journals (Sweden)

    Emiliano Schena

    2017-06-01

    Full Text Available Laser ablation (LA is gaining acceptance for the treatment of tumors as an alternative to surgical resection. This paper reviews the use of lasers for ablative and surgical applications. Also reviewed are solutions aimed at improving LA outcomes: hyperthermal treatment planning tools and thermometric techniques during LA, used to guide the surgeon in the choice and adjustment of the optimal laser settings, and the potential use of nanoparticles to allow biologic selectivity of ablative treatments. Promising technical solutions and a better knowledge of laser-tissue interaction should allow LA to be used in a safe and effective manner as a cancer treatment.

  6. G protein-coupled receptor (GPCR) signaling via heterotrimeric G proteins from endosomes.

    Science.gov (United States)

    Tsvetanova, Nikoleta G; Irannejad, Roshanak; von Zastrow, Mark

    2015-03-13

    Some G protein-coupled receptors (GPCRs), in addition to activating heterotrimeric G proteins in the plasma membrane, appear to elicit a "second wave" of G protein activation after ligand-induced internalization. We briefly summarize evidence supporting this view and then discuss what is presently known about the functional significance of GPCR-G protein activation in endosomes. Endosomal activation can shape the cellular response temporally by prolonging its overall duration, and may shape the response spatially by moving the location of intracellular second messenger production relative to effectors. © 2015 by The American Society for Biochemistry and Molecular Biology, Inc.

  7. Endocytosis of wheat germ agglutinin binding sites from the cell surface into a tubular endosomal network.

    Science.gov (United States)

    Raub, T J; Koroly, M J; Roberts, R M

    1990-04-01

    By using fluorescence and electron microscopy, the endocytic pathway encountered by cell surface components after they had bound wheat germ agglutinin (WGA) was visualized. The majority of these components are thought to consist of sialylated glycoproteins (HMWAG) that represent a subpopulation of the total cell surface proteins but most of the externally disposed plasma membrane proteins of the cell. Examination of semi-thin sections by medium- and high-voltage electron microscopy revealed the three-dimensional organization of vesicular and tubular endosomes. Binding of either fluorescein isothiocyanate-, horseradish peroxidase-, or ferritin-conjugated WGA to cells at 4 degrees C showed that the HMWAG were distributed uniformly over the cell surface. Warming of surface-labeled cells to 37 degrees C resulted in the endocytosis of WGA into peripheral endosomes via invagination of regions of both coated and uncoated membrane. The peripheral endosome appeared as isolated complexes comprising a vesicular element (300-400 nm diam.) surrounded by and continuous with tubular cisternae (45-60 nm diam.), which did not interconnect the endosomes. After 30 min or more label also became localized in a network of anastomosing tubules (45-60 nm diam.) that were located in the centrosomal region of the cell. Endocytosed WGA-HMWAG complexes did not become associated with cisternae of the Golgi apparatus, although tubular and vesicular endosomes were noted in the vicinity of the trans-Golgi region. The accumulation of WGA-HMWAG in the endosomes within the centrosomal region was inhibited when cells were incubated at 18 degrees C. None of these compartments contained acid phosphatase activity, a result that is consistent with other data that the HMWAG do not pass through lysosomes initially. The kinetics of labeling were consistent with the interpretation that recycling of most of the WGA binding surface glycoproteins occurred rapidly from early peripheral endosomes followed by the

  8. GGA1 regulates signal-dependent sorting of BACE1 to recycling endosomes, which moderates Aβ production

    Science.gov (United States)

    Toh, Wei Hong; Chia, Pei Zhi Cheryl; Hossain, Mohammed Iqbal; Gleeson, Paul A.

    2018-01-01

    The diversion of the membrane-bound β-site amyloid precursor protein–(APP) cleaving enzyme (BACE1) from the endolysosomal pathway to recycling endosomes represents an important transport step in the regulation of amyloid beta (Aβ) production. However, the mechanisms that regulate endosome sorting of BACE1 are poorly understood. Here we assessed the transport of BACE1 from early to recycling endosomes and have identified essential roles for the sorting nexin 4 (SNX4)-mediated, signal-independent pathway and for a novel signal-mediated pathway. The signal-mediated pathway is regulated by the phosphorylation of the DXXLL-motif sequence DISLL in the cytoplasmic tail of BACE1. The phosphomimetic S498D BACE1 mutant was trafficked to recycling endosomes at a faster rate compared with wild-type BACE1 or the nonphosphorylatable S498A mutant. The rapid transit of BACE1 S498D from early endosomes was coupled with reduced levels of amyloid precursor protein processing and Aβ production, compared with the S498A mutant. We show that the adaptor, GGA1, and retromer are essential to mediate rapid trafficking of phosphorylated BACE1 to recycling endosomes. In addition, the BACE1 DISLL motif is phosphorylated and regulates endosomal trafficking, in primary neurons. Therefore, post-translational phosphorylation of DISLL enhances the exit of BACE1 from early endosomes, a pathway mediated by GGA1 and retromer, which is important in regulating Aβ production. PMID:29142073

  9. Gαs regulates Glucagon-Like Peptide 1 Receptor-mediated cyclic AMP generation at Rab5 endosomal compartment

    Directory of Open Access Journals (Sweden)

    Shravan Babu Girada

    2017-10-01

    Conclusions: The findings provide the mechanism of endosomal cyclic AMP generation following GLP-1R activation. We identified the specific compartment that serves as an organizing center to generate endosomal cyclic AMP by internalized activated receptor complex.

  10. Anterograde trafficking of KCa3.1 in polarized epithelia is Rab1- and Rab8-dependent and recycling endosome-independent.

    Directory of Open Access Journals (Sweden)

    Claudia A Bertuccio

    Full Text Available The intermediate conductance, Ca2+-activated K+ channel (KCa3.1 targets to the basolateral (BL membrane in polarized epithelia where it plays a key role in transepithelial ion transport. However, there are no studies defining the anterograde and retrograde trafficking of KCa3.1 in polarized epithelia. Herein, we utilize Biotin Ligase Acceptor Peptide (BLAP-tagged KCa3.1 to address these trafficking steps in polarized epithelia, using MDCK, Caco-2 and FRT cells. We demonstrate that KCa3.1 is exclusively targeted to the BL membrane in these cells when grown on filter supports. Following endocytosis, KCa3.1 degradation is prevented by inhibition of lysosomal/proteosomal pathways. Further, the ubiquitylation of KCa3.1 is increased following endocytosis from the BL membrane and PR-619, a deubiquitylase inhibitor, prevents degradation, indicating KCa3.1 is targeted for degradation by ubiquitylation. We demonstrate that KCa3.1 is targeted to the BL membrane in polarized LLC-PK1 cells which lack the μ1B subunit of the AP-1 complex, indicating BL targeting of KCa3.1 is independent of μ1B. As Rabs 1, 2, 6 and 8 play roles in ER/Golgi exit and trafficking of proteins to the BL membrane, we evaluated the role of these Rabs in the trafficking of KCa3.1. In the presence of dominant negative Rab1 or Rab8, KCa3.1 cell surface expression was significantly reduced, whereas Rabs 2 and 6 had no effect. We also co-immunoprecipitated KCa3.1 with both Rab1 and Rab8. These results suggest these Rabs are necessary for the anterograde trafficking of KCa3.1. Finally, we determined whether KCa3.1 traffics directly to the BL membrane or through recycling endosomes in MDCK cells. For these studies, we used either recycling endosome ablation or dominant negative RME-1 constructs and determined that KCa3.1 is trafficked directly to the BL membrane rather than via recycling endosomes. These results are the first to describe the anterograde and retrograde trafficking of KCa3

  11. Actual role of radiofrequency ablation of liver metastases

    Energy Technology Data Exchange (ETDEWEB)

    Pereira, Philippe L. [Eberhard-Karls-University of Tuebingen, Department of Diagnostic Radiology, Tuebingen (Germany)

    2007-08-15

    The liver is, second only to lymph nodes, the most common site for metastatic disease irrespective of the primary tumour. More than 50% of all patients with malignant diseases will develop liver metastases with a significant morbidity and mortality. Although the surgical resection leads to an improved survival in patients with colorectal metastases, only approximately 20% of patients are eligible for surgery. Thermal ablation and especially radiofrequency ablation emerge as an important additional therapy modality for the treatment of liver metastases. RF ablation shows a benefit in life expectancy and may lead in a selected patient group to cure. Percutaneous RF ablation appears safer (versus cryotherapy), easier (versus laser), and more effective (versus ethanol instillation and transarterial chemoembolisation) compared with other minimally invasive procedures. RF ablation can be performed by a percutaneous, laparoscopical or laparotomic approach, and may be potentially combined with chemotherapy and surgery. At present ideal candidates have tumours with a maximum diameter less than 3.5 cm. An untreatable primary tumour or a systemic disease represents contraindications for performing local therapies. Permanent technical improvements of thermal ablation devices and a better integration of thermal ablation in the overall patient care may lead to prognosis improvement in patients with liver metastases. (orig.)

  12. Actual role of radiofrequency ablation of liver metastases

    International Nuclear Information System (INIS)

    Pereira, Philippe L.

    2007-01-01

    The liver is, second only to lymph nodes, the most common site for metastatic disease irrespective of the primary tumour. More than 50% of all patients with malignant diseases will develop liver metastases with a significant morbidity and mortality. Although the surgical resection leads to an improved survival in patients with colorectal metastases, only approximately 20% of patients are eligible for surgery. Thermal ablation and especially radiofrequency ablation emerge as an important additional therapy modality for the treatment of liver metastases. RF ablation shows a benefit in life expectancy and may lead in a selected patient group to cure. Percutaneous RF ablation appears safer (versus cryotherapy), easier (versus laser), and more effective (versus ethanol instillation and transarterial chemoembolisation) compared with other minimally invasive procedures. RF ablation can be performed by a percutaneous, laparoscopical or laparotomic approach, and may be potentially combined with chemotherapy and surgery. At present ideal candidates have tumours with a maximum diameter less than 3.5 cm. An untreatable primary tumour or a systemic disease represents contraindications for performing local therapies. Permanent technical improvements of thermal ablation devices and a better integration of thermal ablation in the overall patient care may lead to prognosis improvement in patients with liver metastases. (orig.)

  13. Cartilage ablation studies using mid-IR free electron laser

    Science.gov (United States)

    Youn, Jong-In; Peavy, George M.; Venugopalan, Vasan

    2005-04-01

    The ablation rate of articular cartilage and fibrocartilage (meniscus), were quantified to examine wavelength and tissue-composition dependence of ablation efficiency for selected mid-infrared wavelengths. The wavelengths tested were 2.9 um (water dominant absorption), 6.1 (protein and water absorption) and 6.45 um (protein dominant absorption) generated by the Free Electron Laser (FEL) at Vanderbilt University. The measurement of tissue mass removal using a microbalance during laser ablation was conducted to determine the ablation rates of cartilage. The technique can be accurate over methods such as profilometer and histology sectioning where tissue surface and the crater morphology may be affected by tissue processing. The ablation efficiency was found to be dependent upon the wavelength. Both articular cartilage and meniscus (fibrocartilage) ablations at 6.1 um were more efficient than those at the other wavelengths evaluated. We observed the lowest ablation efficiency of both types of cartilage with the 6.45 um wavelength, possibly due to the reduction in water absorption at this wavelength in comparison to the other wavelengths that were evaluated.

  14. Intense harmonic generation from various ablation media

    International Nuclear Information System (INIS)

    Ozaki, T.; Elouga, L.; Suzuki, M.; Kuroda, H.; Ganeev, R.A.

    2006-01-01

    lens (f = 680 nm). The high-order harmonics were spectrally resolved using a flat-field grazing-incidence XUV spectrometer with a Hitachi 1200-grooves/mm grating. The XUV spectrum was detected by a microchannel plate with phosphor screen and recorded by a CCD camera. Ablation harmonic experiments were performed with silver and indium targets. We selected silver because of its high conversion efficiency, and indium for its peculiar intensity enhancement effects of the 13 th harmonics. Due to the high intensities of the ablation harmonics, all harmonic spectra were obtained in a single shot. Experiments reveal that the pre-pulse condition for maximum harmonic generation is distinctly different for the two targets. Hydrodynamic simulations using the HYADES code show that the high density of the ablation medium results in strong absorption of the generated harmonics. Therefore, the trade-off between high harmonic efficiency and high absorption is especially important in the present scheme, which can change significantly with the pre-plasma condition. Results with indium targets also reveal a distinct change in the ratio between the 13 th and 15 th harmonic intensity when varying the main pump intensity. This phenomenon is attributed to the change in the resonance conditions of the 13 th harmonic with a strong radiative transition of the In + ion, due to the AC-Stark effect. We will also present new results on ablation harmonics using tin targets.

  15. Neuron specific Rab4 effector GRASP-1 coordinates membrane specialization and maturation of recycling endosomes

    NARCIS (Netherlands)

    C.C. Hoogenraad (Casper); I. Popa (Ioana); K. Futai (Kensuke); E. Sanchez-Martinez (Emma); P. Wulf (Phebe); T. van Vlijmen (Thijs); B.R. Dortland (Bjorn); V. Oorschot (Viola); R. Govers (Robert); M. Monti (Maria); A.J.R. Heck (Albert); M. Sheng (Morgan); J. Klumperman (Judith); H. Rehmann (Holger); D. Jaarsma (Dick); L.C. Kapitein (Lukas); P. van der Sluijs

    2010-01-01

    textabstractThe endosomal pathway in neuronal dendrites is essential for membrane receptor trafficking and proper synaptic function and plasticity. However, the molecular mechanisms that organize specific endocytic trafficking routes are poorly understood. Here, we identify GRIP-associated protein-1

  16. Natural Modulators of Endosomal Toll-Like Receptor-Mediated Psoriatic Skin Inflammation

    Directory of Open Access Journals (Sweden)

    Chao-Yang Lai

    2017-01-01

    Full Text Available Psoriasis is a chronic inflammatory autoimmune disease that can be initiated by excessive activation of endosomal toll-like receptors (TLRs, particularly TLR7, TLR8, and TLR9. Therefore, inhibitors of endosomal TLR activation are being investigated for their ability to treat this disease. The currently approved biological drugs adalimumab, etanercept, infliximab, ustekinumab, ixekizumab, and secukizumab are antibodies against effector cytokines that participate in the initiation and development of psoriasis. Several immune modulatory oligonucleotides and small molecular weight compounds, including IMO-3100, IMO-8400, and CPG-52364, that block the interaction between endosomal TLRs and their ligands are under clinical investigation for their effectiveness in the treatment of psoriasis. In addition, several chemical compounds, including AS-2444697, PF-05387252, PF-05388169, PF-06650833, ML120B, and PHA-408, can inhibit TLR signaling. Although these compounds have demonstrated anti-inflammatory activity in animal models, their therapeutic potential for the treatment of psoriasis has not yet been tested. Recent studies demonstrated that natural compounds derived from plants, fungi, and bacteria, including mustard seed, Antrodia cinnamomea extract, curcumin, resveratrol, thiostrepton, azithromycin, and andrographolide, inhibited psoriasis-like inflammation induced by the TLR7 agonist imiquimod in animal models. These natural modulators employ different mechanisms to inhibit endosomal TLR activation and are administered via different routes. Therefore, they represent candidate psoriasis drugs and might lead to the development of new treatment options.

  17. TLR2 ligands induce NF-κB activation from endosomal compartments of human monocytes.

    Directory of Open Access Journals (Sweden)

    Karim J Brandt

    Full Text Available Localization of Toll-like receptors (TLR in subcellular organelles is a major strategy to regulate innate immune responses. While TLR4, a cell-surface receptor, signals from both the plasma membrane and endosomal compartments, less is known about the functional role of endosomal trafficking upon TLR2 signaling. Here we show that the bacterial TLR2 ligands Pam3CSK4 and LTA activate NF-κB-dependent signaling from endosomal compartments in human monocytes and in a NF-κB sensitive reporter cell line, despite the expression of TLR2 at the cell surface. Further analyses indicate that TLR2-induced NF-κB activation is controlled by a clathrin/dynamin-dependent endocytosis mechanism, in which CD14 serves as an important upstream regulator. These findings establish that internalization of cell-surface TLR2 into endosomal compartments is required for NF-κB activation. These observations further demonstrate the need of endocytosis in the activation and regulation of TLR2-dependent signaling pathways.

  18. Cooperative endocytosis of the endosomal SNARE protein syntaxin-8 and the potassium channel TASK-1

    Science.gov (United States)

    Renigunta, Vijay; Fischer, Thomas; Zuzarte, Marylou; Kling, Stefan; Zou, Xinle; Siebert, Kai; Limberg, Maren M.; Rinné, Susanne; Decher, Niels; Schlichthörl, Günter; Daut, Jürgen

    2014-01-01

    The endosomal SNARE protein syntaxin-8 interacts with the acid-sensitive potassium channel TASK-1. The functional relevance of this interaction was studied by heterologous expression of these proteins (and mutants thereof) in Xenopus oocytes and in mammalian cell lines. Coexpression of syntaxin-8 caused a fourfold reduction in TASK-1 current, a corresponding reduction in the expression of TASK-1 at the cell surface, and a marked increase in the rate of endocytosis of the channel. TASK-1 and syntaxin-8 colocalized in the early endosomal compartment, as indicated by the endosomal markers 2xFYVE and rab5. The stimulatory effect of the SNARE protein on the endocytosis of the channel was abolished when both an endocytosis signal in TASK-1 and an endocytosis signal in syntaxin-8 were mutated. A syntaxin-8 mutant that cannot assemble with other SNARE proteins had virtually the same effect as wild-type syntaxin-8. Total internal reflection fluorescence microscopy showed formation and endocytosis of vesicles containing fluorescence-tagged clathrin, TASK-1, and/or syntaxin-8. Our results suggest that the unassembled form of syntaxin-8 and the potassium channel TASK-1 are internalized via clathrin-mediated endocytosis in a cooperative manner. This implies that syntaxin-8 regulates the endocytosis of TASK-1. Our study supports the idea that endosomal SNARE proteins can have functions unrelated to membrane fusion. PMID:24743596

  19. Snapin-regulated late endosomal transport is critical for efficient autophagy-lysosomal function in neurons.

    Science.gov (United States)

    Cai, Qian; Lu, Li; Tian, Jin-Hua; Zhu, Yi-Bing; Qiao, Haifa; Sheng, Zu-Hang

    2010-10-06

    Neuron maintenance and survival require late endocytic transport from distal processes to the soma where lysosomes are predominantly localized. Here, we report a role for Snapin in attaching dynein to late endosomes through its intermediate chain (DIC). snapin(-/-) neurons exhibit aberrant accumulation of immature lysosomes, clustering and impaired retrograde transport of late endosomes along processes, reduced lysosomal proteolysis due to impaired delivery of internalized proteins and hydrolase precursors from late endosomes to lysosomes, and impaired clearance of autolysosomes, combined with reduced neuron viability and neurodegeneration. The phenotypes are rescued by expressing the snapin transgene, but not the DIC-binding-defective Snapin-L99K mutant. Snapin overexpression in wild-type neurons enhances late endocytic transport and lysosomal function, whereas expressing the mutant defective in Snapin-DIC coupling shows a dominant-negative effect. Altogether, our study highlights new mechanistic insights into how Snapin-DIC coordinates retrograde transport and late endosomal-lysosomal trafficking critical for autophagy-lysosomal function, and thus neuronal homeostasis. Copyright © 2010 Elsevier Inc. All rights reserved.

  20. Endosomal gene expression: a new indicator for prostate cancer patient prognosis?

    LENUS (Irish Health Repository)

    Johnson, Ian R D

    2015-11-10

    Prostate cancer continues to be a major cause of morbidity and mortality in men, but a method for accurate prognosis in these patients is yet to be developed. The recent discovery of altered endosomal biogenesis in prostate cancer has identified a fundamental change in the cell biology of this cancer, which holds great promise for the identification of novel biomarkers that can predict disease outcomes. Here we have identified significantly altered expression of endosomal genes in prostate cancer compared to non-malignant tissue in mRNA microarrays and confirmed these findings by qRT-PCR on fresh-frozen tissue. Importantly, we identified endosomal gene expression patterns that were predictive of patient outcomes. Two endosomal tri-gene signatures were identified from a previously published microarray cohort and had a significant capacity to stratify patient outcomes. The expression of APPL1, RAB5A, EEA1, PDCD6IP, NOX4 and SORT1 were altered in malignant patient tissue, when compared to indolent and normal prostate tissue. These findings support the initiation of a case-control study using larger cohorts of prostate tissue, with documented patient outcomes, to determine if different combinations of these new biomarkers can accurately predict disease status and clinical progression in prostate cancer patients.

  1. Integrin-linked kinase and ELMO2 modulate recycling endosomes in keratinocytes.

    Science.gov (United States)

    Ho, Ernest; Ivanova, Iordanka A; Dagnino, Lina

    2016-12-01

    The formation of tight cell-cell junctions is essential in the epidermis for its barrier properties. In this tissue, keratinocytes follow a differentiation program tightly associated with their movement from the innermost basal to the outer suprabasal layers, and with changes in their cell-cell adhesion profile. Intercellular adhesion in keratinocytes is mediated through cell-cell contacts, including E-cadherin-based adherens junctions. Although the mechanisms that mediate E-cadherin delivery to the plasma membrane have been widely studied in simple epithelia, this process is less well understood in the stratified epidermis. In this study, we have investigated the role of Engulfment and Cell Motility 2 (ELMO2) and integrin-linked kinase (ILK) in the positioning of E-cadherin-containing recycling endosomes during establishment of cell-cell contacts in differentiating keratinocytes. We now show that induction of keratinocyte differentiation by Ca 2+ is accompanied by localization of ELMO2 and ILK to Rab4- and Rab11a-containing recycling endosomes. The positioning of long-loop Rab11a-positive endosomes at areas adjacent to cell-cell contacts is disrupted in ELMO2- or ILK-deficient keratinocytes, and is associated with impaired localization of E-cadherin to cell borders. Our studies show a previously unrecognized role for ELMO2 and ILK in modulation of endosomal positioning, which may play key roles in epidermal sheet maintenance and permeability barrier function. Copyright © 2016 Elsevier B.V. All rights reserved.

  2. Rab11-FIP3 Regulation of Lck Endosomal Traffic Controls TCR Signal Transduction.

    Science.gov (United States)

    Bouchet, Jérôme; Del Río-Iñiguez, Iratxe; Vázquez-Chávez, Elena; Lasserre, Rémi; Agüera-González, Sonia; Cuche, Céline; McCaffrey, Mary W; Di Bartolo, Vincenzo; Alcover, Andrés

    2017-04-01

    The role of endosomes in receptor signal transduction is a long-standing question, which remains largely unanswered. The T cell Ag receptor and various components of its proximal signaling machinery are associated with distinct endosomal compartments, but how endosomal traffic affects T cell signaling remains ill-defined. In this article, we demonstrate in human T cells that the subcellular localization and function of the protein tyrosine kinase Lck depends on the Rab11 effector FIP3 (Rab11 family interacting protein-3). FIP3 overexpression or silencing and its ability to interact with Rab11 modify Lck subcellular localization and its delivery to the immunological synapse. Importantly, FIP3-dependent Lck localization controls early TCR signaling events, such as tyrosine phosphorylation of TCRζ, ZAP70, and LAT and intracellular calcium concentration, as well as IL-2 gene expression. Interestingly, FIP3 controls both steady-state and poststimulation phosphotyrosine and calcium levels. Finally, our findings indicate that FIP3 modulates TCR-CD3 cell surface expression via the regulation of steady-state Lck-mediated TCRζ phosphorylation, which in turn controls TCRζ protein levels. This may influence long-term T cell activation in response to TCR-CD3 stimulation. Therefore, our data underscore the importance of finely regulated endosomal traffic in TCR signal transduction and T cell activation leading to IL-2 production. Copyright © 2017 by The American Association of Immunologists, Inc.

  3. Lesion size in relation to ablation site during radiofrequency ablation

    DEFF Research Database (Denmark)

    Petersen, H H; Chen, X; Pietersen, A

    1998-01-01

    This study was designed to investigate the effect of the convective cooling of the tip of the ablation electrode during temperature controlled radiofrequency ablation. In vivo two different application sites in the left ventricle of anaesthetised pigs were ablated and in vitro ablation was perfor......This study was designed to investigate the effect of the convective cooling of the tip of the ablation electrode during temperature controlled radiofrequency ablation. In vivo two different application sites in the left ventricle of anaesthetised pigs were ablated and in vitro ablation...... was performed during two different flow-velocities in a tissue bath, while electrode contact pressure and position were unchanged. Target temperature was 80 degrees C. Obtained tip temperature, power consumption and lesion dimensions were measured. In vivo lesion volume, depth and width were found significantly.......61 in vitro). We conclude that during temperature controlled radiofrequency ablation lesion size differs for septal and apical left ventricular applications. Differences in convective cooling might play an important role in this respect. This is supported by our in vitro experiments, where increased...

  4. The V-ATPase a2-subunit as a putative endosomal pH-sensor.

    Science.gov (United States)

    Marshansky, V

    2007-11-01

    V-ATPase (vesicular H(+)-ATPase)-driven intravesicular acidification is crucial for vesicular trafficking. Defects in vesicular acidification and trafficking have recently been recognized as essential determinants of various human diseases. An important role of endosomal acidification in receptor-ligand dissociation and in activation of lysosomal hydrolytic enzymes is well established. However, the molecular mechanisms by which luminal pH information is transmitted to the cytosolic small GTPases that control trafficking events such as budding, coat formation and fusion are unknown. Here, we discuss our recent discovery that endosomal V-ATPase is a pH-sensor regulating the degradative pathway. According to our model, V-ATPase is responsible for: (i) the generation of a pH gradient between vesicular membranes; (ii) sensing of intravesicular pH; and (iii) transmitting this information to the cytosolic side of the membrane. We also propose the hypothetical molecular mechanism involved in function of the V-ATPase a2-subunit as a putative pH-sensor. Based on extensive experimental evidence on the crucial role of histidine residues in the function of PSPs (pH-sensing proteins) in eukaryotic cells, we hypothesize that pH-sensitive histidine residues within the intra-endosomal loops and/or C-terminal luminal tail of the a2-subunit could also be involved in the pH-sensing function of V-ATPase. However, in order to identify putative pH-sensitive histidine residues and to test this hypothesis, it is absolutely essential that we increase our understanding of the folding and transmembrane topology of the a-subunit isoforms of V-ATPase. Thus the crucial role of intra-endosomal histidine residues in pH-dependent conformational changes of the V-ATPase a2-isoform, its interaction with cytosolic small GTPases and ultimately in its acidification-dependent regulation of the endosomal/lysosomal protein degradative pathway remain to be determined.

  5. Photoactive dye enhanced tissue ablation for endoscopic laser prostatectomy

    Science.gov (United States)

    Ahn, Minwoo; Nguyen, Trung Hau; Nguyen, Van Phuc; Oh, Junghwan; Kang, Hyun Wook

    2015-02-01

    Laser light has been widely used as a surgical tool to treat benign prostate hyperplasia with high laser power. The purpose of this study was to validate the feasibility of photoactive dye injection to enhance light absorption and eventually to facilitate tissue ablation with low laser power. The experiment was implemented on chicken breast due to minimal optical absorption Amaranth (AR), black dye (BD), hemoglobin powder (HP), and endoscopic marker (EM), were selected and tested in vitro with a customized 532-nm laser system with radiant exposure ranging from 0.9 to 3.9 J/cm2. Light absorbance and ablation threshold were measured with UV-VIS spectrometer and Probit analysis, respectively, and compared to feature the function of the injected dyes. Ablation performance with dye-injection was evaluated in light of radiant exposure, dye concentration, and number of injection. Higher light absorption by injected dyes led to lower ablation threshold as well as more efficient tissue removal in the order of AR, BD, HP, and EM. Regardless of the injected dyes, ablation efficiency principally increased with input parameter. Among the dyes, AR created the highest ablation rate of 44.2+/-0.2 μm/pulse due to higher absorbance and lower ablation threshold. Preliminary tests on canine prostate with a hydraulic injection system demonstrated that 80 W with dye injection yielded comparable ablation efficiency to 120 W with no injection, indicating 33 % reduced laser power with almost equivalent performance. In-depth comprehension on photoactive dye-enhanced tissue ablation can help accomplish efficient and safe laser treatment for BPH with low power application.

  6. Field enhancement induced laser ablation

    DEFF Research Database (Denmark)

    Fiutowski, Jacek; Maibohm, Christian; Kjelstrup-Hansen, Jakob

    Sub-diffraction spatially resolved, quantitative mapping of strongly localized field intensity enhancement on gold nanostructures via laser ablation of polymer thin films is reported. Illumination using a femtosecond laser scanning microscope excites surface plasmons in the nanostructures....... The accompanying field enhancement substantially lowers the ablation threshold of the polymer film and thus creates local ablation spots and corresponding topographic modifications of the polymer film. Such modifications are quantified straightforwardly via scanning electron and atomic force microscopy. Thickness...

  7. Micro Sampling System for Highly Radioactive Specimen by Laser Ablation

    Energy Technology Data Exchange (ETDEWEB)

    Han, Sun Ho; Ha, Yeong Keong; Han, Ki Chul; Park, Yang Soon; Jee, Kwang Yong; Kim, Won Ho [Korea Atomic Energy Research Institute, Daejeon (Korea, Republic of)

    2005-03-15

    Shielded laser ablation system composed of laser system, image analyser, XYZ translator with motion controller, ablation chamber, manipulator and various optics was designed. Nd:YAG laser which can be tunable from 1064 nm to 266 nm was selected as light source. CCD camera(< x 200) was chosen to analyze a crater less than 50 un in diameter. XYZ translator was composed of three linear stage which can travel 50 w with a minimum movement of 1 {mu}m and motion controller. Before the performance test, each part of system was optically aligned. To perform the ablation test, the specimen was ablated by 50 {mu}m interval and observed by image analyser The shape of crater was almost round, indicating laser beam has homogeneous energy distribution. The resolution and magnification of image system were compatible with the design.

  8. The late endosome/lysosome-anchored p18-mTORC1 pathway controls terminal maturation of lysosomes

    Energy Technology Data Exchange (ETDEWEB)

    Takahashi, Yusuke; Nada, Shigeyuki; Mori, Shunsuke; Soma-Nagae, Taeko; Oneyama, Chitose [Department of Oncogene Research, Research Institute for Microbial Diseases, Osaka University, 3-1 Yamadaoka, Suita, Osaka 565-0871 (Japan); Okada, Masato, E-mail: okadam@biken.osaka-u.ac.jp [Department of Oncogene Research, Research Institute for Microbial Diseases, Osaka University, 3-1 Yamadaoka, Suita, Osaka 565-0871 (Japan)

    2012-01-27

    Highlights: Black-Right-Pointing-Pointer p18 is a membrane adaptor that anchors mTORC1 to late endosomes/lysosomes. Black-Right-Pointing-Pointer We examine the role of the p18-mTORC1 pathway in lysosome biogenesis. Black-Right-Pointing-Pointer The loss of p18 causes accumulation of intact late endosomes by arresting lysosome maturation. Black-Right-Pointing-Pointer Inhibition of mTORC1 activity with rapamycin phenocopies the defects of p18 loss. Black-Right-Pointing-Pointer The p18-mTORC1 pathway plays crucial roles in the terminal maturation of lysosomes. -- Abstract: The late endosome/lysosome membrane adaptor p18 (or LAMTOR1) serves as an anchor for the mammalian target of rapamycin complex 1 (mTORC1) and is required for its activation on lysosomes. The loss of p18 causes severe defects in cell growth as well as endosome dynamics, including membrane protein transport and lysosome biogenesis. However, the mechanisms underlying these effects on lysosome biogenesis remain unknown. Here, we show that the p18-mTORC1 pathway is crucial for terminal maturation of lysosomes. The loss of p18 causes aberrant intracellular distribution and abnormal sizes of late endosomes/lysosomes and an accumulation of late endosome specific components, including Rab7, RagC, and LAMP1; this suggests that intact late endosomes accumulate in the absence of p18. These defects are phenocopied by inhibiting mTORC1 activity with rapamycin. Loss of p18 also suppresses the integration of late endosomes and lysosomes, resulting in the defective degradation of tracer proteins. These results suggest that the p18-mTORC1 pathway plays crucial roles in the late stages of lysosomal maturation, potentially in late endosome-lysosome fusion, which is required for processing of various macromolecules.

  9. Human ClC-6 is a late endosomal glycoprotein that associates with detergent-resistant lipid domains.

    Directory of Open Access Journals (Sweden)

    Sofie Ignoul

    Full Text Available BACKGROUND: The mammalian CLC protein family comprises nine members (ClC-1 to -7 and ClC-Ka, -Kb that function either as plasma membrane chloride channels or as intracellular chloride/proton antiporters, and that sustain a broad spectrum of cellular processes, such as membrane excitability, transepithelial transport, endocytosis and lysosomal degradation. In this study we focus on human ClC-6, which is structurally most related to the late endosomal/lysomal ClC-7. PRINCIPAL FINDINGS: Using a polyclonal affinity-purified antibody directed against a unique epitope in the ClC-6 COOH-terminal tail, we show that human ClC-6, when transfected in COS-1 cells, is N-glycosylated in a region that is evolutionary poorly conserved between mammalian CLC proteins and that is located between the predicted helices K and M. Three asparagine residues (N410, N422 and N432 have been defined by mutagenesis as acceptor sites for N-glycosylation, but only two of the three sites seem to be simultaneously N-glycosylated. In a differentiated human neuroblastoma cell line (SH-SY5Y, endogenous ClC-6 colocalizes with LAMP-1, a late endosomal/lysosomal marker, but not with early/recycling endosomal markers such as EEA-1 and transferrin receptor. In contrast, when transiently expressed in COS-1 or HeLa cells, human ClC-6 mainly overlaps with markers for early/recycling endosomes (transferrin receptor, EEA-1, Rab5, Rab4 and not with late endosomal/lysosomal markers (LAMP-1, Rab7. Analogously, overexpression of human ClC-6 in SH-SY5Y cells also leads to an early/recycling endosomal localization of the exogenously expressed ClC-6 protein. Finally, in transiently transfected COS-1 cells, ClC-6 copurifies with detergent-resistant membrane fractions, suggesting its partitioning in lipid rafts. Mutating a juxtamembrane string of basic amino acids (amino acids 71-75: KKGRR disturbs the association with detergent-resistant membrane fractions and also affects the segregation of ClC-6

  10. Radiofrequency ablation of osteoid osteoma

    NARCIS (Netherlands)

    Vanderschueren, Geert Maria Joris Michael

    2009-01-01

    The main purpose of this thesis was to evaluate the effectiveness and safety of CT-guided radiofrequency ablation for the treatment of spinal and non-spinal osteoid osteomas. Furthermore, the technical requirements needed for safe radiofrequency ablation and the clinical outcome after radiofrequency

  11. [Radiofrequency ablation of hepatocellular carcinoma].

    Science.gov (United States)

    Widmann, Gerlig; Schullian, Peter; Bale, Reto

    2013-03-01

    Percutaneous radiofrequency ablation (RFA) is well established in the treatment of hepatocellular carcinoma (HCC). Due to its curative potential, it is the method of choice for non resectable BCLC (Barcelona Liver Clinic) 0 and A. RFA challenges surgical resection for small HCC and is the method of choice in bridging for transplantation and recurrence after resection or transplantation. The technical feasibility of RFA depends on the size and location of the HCC and the availability of ablation techniques (one needle techniques, multi-needle techniques). More recently, stereotactic multi-needle techniques with 3D trajectory planning and guided needle placement substantially improve the spectrum of treatable lesions including large volume tumors. Treatment success depends on the realization of ablations with large intentional margins of tumor free tissue (A0 ablation in analogy to R0 resection), which has to be documented by fusion of post- with pre-ablation images, and confirmed during follow-up imaging.

  12. Laboratory Simulations of Micrometeoroid Ablation

    Science.gov (United States)

    Thomas, Evan Williamson

    Each day, several tons of meteoric material enters Earth's atmosphere, the majority of which consist of small dust particles (micrometeoroids) that completely ablate at high altitudes. The dust input has been suggested to play a role in a variety of phenomena including: layers of metal atoms and ions, nucleation of noctilucent clouds, effects on stratospheric aerosols and ozone chemistry, and the fertilization of the ocean with bio-available iron. Furthermore, a correct understanding of the dust input to the Earth provides constraints on inner solar system dust models. Various methods are used to measure the dust input to the Earth including satellite detectors, radar, lidar, rocket-borne detectors, ice core and deep-sea sediment analysis. However, the best way to interpret each of these measurements is uncertain, which leads to large uncertainties in the total dust input. To better understand the ablation process, and thereby reduce uncertainties in micrometeoroid ablation measurements, a facility has been developed to simulate the ablation of micrometeoroids in laboratory conditions. An electrostatic dust accelerator is used to accelerate iron particles to relevant meteoric velocities (10-70 km/s). The particles are then introduced into a chamber pressurized with a target gas, and they partially or completely ablate over a short distance. An array of diagnostics then measure, with timing and spatial resolution, the charge and light that is generated in the ablation process. In this thesis, we present results from the newly developed ablation facility. The ionization coefficient, an important parameter for interpreting meteor radar measurements, is measured for various target gases. Furthermore, experimental ablation measurements are compared to predictions from commonly used ablation models. In light of these measurements, implications to the broader context of meteor ablation are discussed.

  13. Fabrication of CVD graphene-based devices via laser ablation for wafer-scale characterization

    DEFF Research Database (Denmark)

    Mackenzie, David; Buron, Jonas Christian Due; Whelan, Patrick Rebsdorf

    2015-01-01

    Selective laser ablation of a wafer-scale graphene film is shown to provide flexible, high speed (1 wafer/hour) device fabrication while avoiding the degradation of electrical properties associated with traditional lithographic methods. Picosecond laser pulses with single pulse peak fluences of 140......-effect mobility, doping level, on–off ratio, and conductance minimum before and after laser ablation fabrication....

  14. Plasma luminescence feedback control system for precise ultrashort pulse laser tissue ablation

    Science.gov (United States)

    Kim, Beop-Min; Feit, Michael D.; Rubenchik, Alexander M.; Gold, David M.; Darrow, Christopher B.; Marion, John E., II; Da Silva, Luiz B.

    1998-05-01

    Plasma luminescence spectroscopy was used for precise ablation of bone tissue without damaging nearby soft tissue using an ultrashort pulse laser. Strong contrast of the luminescence spectra between bone marrow and spinal cord provided the real time feedback control so bone tissue is selectively ablated while preserving the spinal cord.

  15. LMTK1 regulates dendritic formation by regulating movement of Rab11A-positive endosomes.

    Science.gov (United States)

    Takano, Tetsuya; Urushibara, Tomoki; Yoshioka, Nozomu; Saito, Taro; Fukuda, Mitsunori; Tomomura, Mineko; Hisanaga, Shin-Ichi

    2014-06-01

    Neurons extend two types of neurites-axons and dendrites-that differ in structure and function. Although it is well understood that the cytoskeleton plays a pivotal role in neurite differentiation and extension, the mechanisms by which membrane components are supplied to growing axons or dendrites is largely unknown. We previously reported that the membrane supply to axons is regulated by lemur kinase 1 (LMTK1) through Rab11A-positive endosomes. Here we investigate the role of LMTK1 in dendrite formation. Down-regulation of LMTK1 increases dendrite growth and branching of cerebral cortical neurons in vitro and in vivo. LMTK1 knockout significantly enhances the prevalence, velocity, and run length of anterograde movement of Rab11A-positive endosomes to levels similar to those expressing constitutively active Rab11A-Q70L. Rab11A-positive endosome dynamics also increases in the cell body and growth cone of LMTK1-deficient neurons. Moreover, a nonphosphorylatable LMTK1 mutant (Ser34Ala, a Cdk5 phosphorylation site) dramatically promotes dendrite growth. Thus LMTK1 negatively controls dendritic formation by regulating Rab11A-positive endosomal trafficking in a Cdk5-dependent manner, indicating the Cdk5-LMTK1-Rab11A pathway as a regulatory mechanism of dendrite development as well as axon outgrowth. © 2014 Takano et al. This article is distributed by The American Society for Cell Biology under license from the author(s). Two months after publication it is available to the public under an Attribution–Noncommercial–Share Alike 3.0 Unported Creative Commons License (http://creativecommons.org/licenses/by-nc-sa/3.0).

  16. Endosomal protein sorting and autophagy genes contribute to the regulation of yeast life span.

    Science.gov (United States)

    Longo, Valter D; Nislow, Corey; Fabrizio, Paola

    2010-11-01

    Accumulating evidence from various organisms points to a role for autophagy in the regulation of life span. By performing a genome-wide screen to identify novel life span determinants in Saccharomyces cerevisiae, we have obtained further insights into the autophagy-related and -unrelated degradation processes that may be important for preventing cellular senescence. The generation of multivesicular bodies and their fusion with the vacuole in the endosomal pathway emerged as novel cell functions involved in yeast chronological survival and longevity extension.

  17. Endosome-based protein trafficking and Ca2+ homeostasis in the heart

    Directory of Open Access Journals (Sweden)

    Jerry eCurran

    2015-02-01

    Full Text Available The ability to dynamically regulate, traffic, retain, and recycle proteins within the cell membrane is fundamental to life and central to the normal function of the heart and cardiovascular system. In the heart, these systems are essential for the regulation of cardiac calcium, both at the level of the plasma membrane, but also at local domains of the endoplasmic reticulum, sarcoplasmic reticulum, mitochondria, nucleus, and nuclear envelope. One intracellular pathway often overlooked in relation to cardiovascular calcium regulation and signaling is the endosome-based trafficking pathway. Highlighting its importance, this system and its molecular components are evolutionarily conserved across all metazoans. However, remarkably little is known of how endosome-based protein trafficking and recycling functions within mammalian cells systems, especially in the heart. The vast majority of what is known has been derived from heterologous cell systems. However, recently, more appropriate cell and animal models been developed that have allowed researchers to begin to understand how this system functions within the intact physiological environment. All excitable cells, including cardiomyocytes, depend on the proper expression and organization of multiple ion channels, pumps, exchangers, and transporters within the plasma membrane. As the endosomal system acts to regulate the expression and localization of membrane proteins, understanding the in vivo function of this system in the heart is important. This review will focus on endosome-based protein trafficking in the heart in both health and disease. Special emphasis will be given to the role played by the family of endocytic regulatory proteins, C-terminal Eps15 homology domain -containing proteins (EHDs, as recent data demonstrates that this family of proteins is essential for the proper trafficking and localization and of key proteins involved in excitation-contraction coupling.

  18. The Endosome-associated Deubiquitinating Enzyme USP8 Regulates BACE1 Enzyme Ubiquitination and Degradation.

    Science.gov (United States)

    Yeates, Eniola Funmilayo Aduke; Tesco, Giuseppina

    2016-07-22

    The β-site amyloid precursor protein-cleaving enzyme (BACE1) is the rate-limiting enzyme in the production of amyloid-β, the toxic peptide that accumulates in the brain of subjects affected by Alzheimer disease. Our previous studies have shown that BACE1 is degraded via the lysosomal pathway and that that depletion of the trafficking molecule Golgi-localized γ-ear-containing ARF-binding protein 3 (GGA3) results in increased BACE1 levels and activity because of impaired lysosomal degradation. We also determined that GGA3 regulation of BACE1 levels requires its ability to bind ubiquitin. Accordingly, we reported that BACE1 is ubiquitinated at lysine 501 and that lack of ubiquitination at lysine 501 produces BACE1 stabilization. Ubiquitin conjugation is a reversible process mediated by deubiquitinating enzymes. The ubiquitin-specific peptidase 8 (USP8), an endosome-associated deubiquitinating enzyme, regulates the ubiquitination, trafficking, and lysosomal degradation of several plasma membrane proteins. Here, we report that RNAi-mediated depletion of USP8 reduced levels of both ectopically expressed and endogenous BACE1 in H4 human neuroglioma cells. Moreover, USP8 depletion increased BACE1 ubiquitination, promoted BACE1 accumulation in the early endosomes and late endosomes/lysosomes, and decreased levels of BACE1 in the recycling endosomes. We also found that decreased BACE1 protein levels were accompanied by a decrease in BACE1-mediated amyloid precursor protein cleavage and amyloid-β levels. Our findings demonstrate that USP8 plays a key role in the trafficking and degradation of BACE1 by deubiquitinating lysine 501. These studies suggest that therapies able to accelerate BACE1 degradation (e.g. by increasing BACE1 ubiquitination) may represent a potential treatment for Alzheimer disease. © 2016 by The American Society for Biochemistry and Molecular Biology, Inc.

  19. The Endosome-associated Deubiquitinating Enzyme USP8 Regulates BACE1 Enzyme Ubiquitination and Degradation*

    Science.gov (United States)

    Yeates, Eniola Funmilayo Aduke; Tesco, Giuseppina

    2016-01-01

    The β-site amyloid precursor protein-cleaving enzyme (BACE1) is the rate-limiting enzyme in the production of amyloid-β, the toxic peptide that accumulates in the brain of subjects affected by Alzheimer disease. Our previous studies have shown that BACE1 is degraded via the lysosomal pathway and that that depletion of the trafficking molecule Golgi-localized γ-ear-containing ARF-binding protein 3 (GGA3) results in increased BACE1 levels and activity because of impaired lysosomal degradation. We also determined that GGA3 regulation of BACE1 levels requires its ability to bind ubiquitin. Accordingly, we reported that BACE1 is ubiquitinated at lysine 501 and that lack of ubiquitination at lysine 501 produces BACE1 stabilization. Ubiquitin conjugation is a reversible process mediated by deubiquitinating enzymes. The ubiquitin-specific peptidase 8 (USP8), an endosome-associated deubiquitinating enzyme, regulates the ubiquitination, trafficking, and lysosomal degradation of several plasma membrane proteins. Here, we report that RNAi-mediated depletion of USP8 reduced levels of both ectopically expressed and endogenous BACE1 in H4 human neuroglioma cells. Moreover, USP8 depletion increased BACE1 ubiquitination, promoted BACE1 accumulation in the early endosomes and late endosomes/lysosomes, and decreased levels of BACE1 in the recycling endosomes. We also found that decreased BACE1 protein levels were accompanied by a decrease in BACE1-mediated amyloid precursor protein cleavage and amyloid-β levels. Our findings demonstrate that USP8 plays a key role in the trafficking and degradation of BACE1 by deubiquitinating lysine 501. These studies suggest that therapies able to accelerate BACE1 degradation (e.g. by increasing BACE1 ubiquitination) may represent a potential treatment for Alzheimer disease. PMID:27302062

  20. Involvement of the endosomal-lysosomal system correlates with regional pathology in Creutzfeldt-Jakob disease

    DEFF Research Database (Denmark)

    Kovács, Gábor G; Gelpi, Ellen; Ströbel, Thomas

    2007-01-01

    The endosomal-lysosomal system (ELS) has been suggested to play a role in the pathogenesis of prion diseases. The purpose of this study was to examine how experimental observations can be translated to human neuropathology and whether alterations of the ELS relate to neuropathologic changes...... correlate with regional pathology. Overloading of this system might impair the function of lysosomal enzymes and thus may mimic some features of lysosomal storage disorders. Udgivelsesdato: 2007-Jul...

  1. Phosphatidylinositol 3,5-Bisphosphate-Rich Membrane Domains in Endosomes and Lysosomes.

    Science.gov (United States)

    Takatori, Sho; Tatematsu, Tsuyako; Cheng, Jinglei; Matsumoto, Jun; Akano, Takuya; Fujimoto, Toyoshi

    2016-02-01

    Phosphatidylinositol 3,5-bisphosphate (PtdIns(3,5)P2 ) has critical functions in endosomes and lysosomes. We developed a method to define nanoscale distribution of PtdIns(3,5)P2 using freeze-fracture electron microscopy. GST-ATG18-4×FLAG was used to label PtdIns(3,5)P2 and its binding to phosphatidylinositol 3-phosphate (PtdIns(3)P) was blocked by an excess of the p40(phox) PX domain. In yeast exposed to hyperosmotic stress, PtdIns(3,5)P2 was concentrated in intramembrane particle (IMP)-deficient domains in the vacuolar membrane, which made close contact with adjacent membranes. The IMP-deficient domain was also enriched with PtdIns(3)P, but was deficient in Vph1p, a liquid-disordered domain marker. In yeast lacking either PtdIns(3,5)P2 or its effector, Atg18p, the IMP-deficient, PtdIns(3)P-rich membranes were folded tightly to make abnormal tubular structures, thus showing where the vacuolar fragmentation process is arrested when PtdIns(3,5)P2 metabolism is defective. In HeLa cells, PtdIns(3,5)P2 was significantly enriched in the vesicular domain of RAB5- and RAB7-positive endosome/lysosomes of the tubulo-vesicular morphology. This biased distribution of PtdIns(3,5)P2 was also observed using fluorescence microscopy, which further showed enrichment of a retromer component, VPS35, in the tubular domain. This is the first report to show segregation of PtdIns(3,5)P2 -rich and -deficient domains in endosome/lysosomes, which should be important for endosome/lysosome functionality. © 2015 John Wiley & Sons A/S. Published by John Wiley & Sons Ltd.

  2. The late endosome/lysosome-anchored p18-mTORC1 pathway controls terminal maturation of lysosomes

    International Nuclear Information System (INIS)

    Takahashi, Yusuke; Nada, Shigeyuki; Mori, Shunsuke; Soma-Nagae, Taeko; Oneyama, Chitose; Okada, Masato

    2012-01-01

    Highlights: ► p18 is a membrane adaptor that anchors mTORC1 to late endosomes/lysosomes. ► We examine the role of the p18-mTORC1 pathway in lysosome biogenesis. ► The loss of p18 causes accumulation of intact late endosomes by arresting lysosome maturation. ► Inhibition of mTORC1 activity with rapamycin phenocopies the defects of p18 loss. ► The p18-mTORC1 pathway plays crucial roles in the terminal maturation of lysosomes. -- Abstract: The late endosome/lysosome membrane adaptor p18 (or LAMTOR1) serves as an anchor for the mammalian target of rapamycin complex 1 (mTORC1) and is required for its activation on lysosomes. The loss of p18 causes severe defects in cell growth as well as endosome dynamics, including membrane protein transport and lysosome biogenesis. However, the mechanisms underlying these effects on lysosome biogenesis remain unknown. Here, we show that the p18-mTORC1 pathway is crucial for terminal maturation of lysosomes. The loss of p18 causes aberrant intracellular distribution and abnormal sizes of late endosomes/lysosomes and an accumulation of late endosome specific components, including Rab7, RagC, and LAMP1; this suggests that intact late endosomes accumulate in the absence of p18. These defects are phenocopied by inhibiting mTORC1 activity with rapamycin. Loss of p18 also suppresses the integration of late endosomes and lysosomes, resulting in the defective degradation of tracer proteins. These results suggest that the p18-mTORC1 pathway plays crucial roles in the late stages of lysosomal maturation, potentially in late endosome–lysosome fusion, which is required for processing of various macromolecules.

  3. Molecular determinants of the interaction between Doa1 and Hse1 involved in endosomal sorting.

    Science.gov (United States)

    Han, Seungsu; Shin, Donghyuk; Choi, Hoon; Lee, Sangho

    2014-03-28

    Yeast Doa1/Ufd3 is an adaptor protein for Cdc48 (p97 in mammal), an AAA type ATPase associated with endoplasmic reticulum-associated protein degradation pathway and endosomal sorting into multivesicular bodies. Doa1 functions in the endosomal sorting by its association with Hse1, a component of endosomal sorting complex required for transport (ESCRT) system. The association of Doa1 with Hse1 was previously reported to be mediated between PFU domain of Doa1 and SH3 of Hse1. However, it remains unclear which residues are specifically involved in the interaction. Here we report that Doa1/PFU interacts with Hse1/SH3 with a moderate affinity of 5 μM. Asn-438 of Doa1/PFU and Trp-254 of Hse1/SH3 are found to be critical in the interaction while Phe-434, implicated in ubiquitin binding via a hydrophobic interaction, is not. Small-angle X-ray scattering measurements combined with molecular docking and biochemical analysis yield the solution structure of the Doa1/PFU:Hse1/SH3 complex. Taken together, our results suggest that hydrogen bonding is a major determinant in the interaction of Doa1/PFU with Hse1/SH3. Copyright © 2014 Elsevier Inc. All rights reserved.

  4. dOCRL maintains immune cell quiescence by regulating endosomal traffic.

    Directory of Open Access Journals (Sweden)

    Steven J Del Signore

    2017-10-01

    Full Text Available Lowe Syndrome is a developmental disorder characterized by eye, kidney, and neurological pathologies, and is caused by mutations in the phosphatidylinositol-5-phosphatase OCRL. OCRL plays diverse roles in endocytic and endolysosomal trafficking, cytokinesis, and ciliogenesis, but it is unclear which of these cellular functions underlie specific patient symptoms. Here, we show that mutation of Drosophila OCRL causes cell-autonomous activation of hemocytes, which are macrophage-like cells of the innate immune system. Among many cell biological defects that we identified in docrl mutant hemocytes, we pinpointed the cause of innate immune cell activation to reduced Rab11-dependent recycling traffic and concomitantly increased Rab7-dependent late endosome traffic. Loss of docrl amplifies multiple immune-relevant signals, including Toll, Jun kinase, and STAT, and leads to Rab11-sensitive mis-sorting and excessive secretion of the Toll ligand Spåtzle. Thus, docrl regulation of endosomal traffic maintains hemocytes in a poised, but quiescent state, suggesting mechanisms by which endosomal misregulation of signaling may contribute to symptoms of Lowe syndrome.

  5. ABMA, a small molecule that inhibits intracellular toxins and pathogens by interfering with late endosomal compartments.

    Science.gov (United States)

    Wu, Yu; Pons, Valérie; Goudet, Amélie; Panigai, Laetitia; Fischer, Annette; Herweg, Jo-Ana; Kali, Sabrina; Davey, Robert A; Laporte, Jérôme; Bouclier, Céline; Yousfi, Rahima; Aubenque, Céline; Merer, Goulven; Gobbo, Emilie; Lopez, Roman; Gillet, Cynthia; Cojean, Sandrine; Popoff, Michel R; Clayette, Pascal; Le Grand, Roger; Boulogne, Claire; Tordo, Noël; Lemichez, Emmanuel; Loiseau, Philippe M; Rudel, Thomas; Sauvaire, Didier; Cintrat, Jean-Christophe; Gillet, Daniel; Barbier, Julien

    2017-11-14

    Intracellular pathogenic microorganisms and toxins exploit host cell mechanisms to enter, exert their deleterious effects as well as hijack host nutrition for their development. A potential approach to treat multiple pathogen infections and that should not induce drug resistance is the use of small molecules that target host components. We identified the compound 1-adamantyl (5-bromo-2-methoxybenzyl) amine (ABMA) from a cell-based high throughput screening for its capacity to protect human cells and mice against ricin toxin without toxicity. This compound efficiently protects cells against various toxins and pathogens including viruses, intracellular bacteria and parasite. ABMA provokes Rab7-positive late endosomal compartment accumulation in mammalian cells without affecting other organelles (early endosomes, lysosomes, the Golgi apparatus, the endoplasmic reticulum or the nucleus). As the mechanism of action of ABMA is restricted to host-endosomal compartments, it reduces cell infection by pathogens that depend on this pathway to invade cells. ABMA may represent a novel class of broad-spectrum compounds with therapeutic potential against diverse severe infectious diseases.

  6. Role of the AP-5 adaptor protein complex in late endosome-to-Golgi retrieval.

    Directory of Open Access Journals (Sweden)

    Jennifer Hirst

    2018-01-01

    Full Text Available The AP-5 adaptor protein complex is presumed to function in membrane traffic, but so far nothing is known about its pathway or its cargo. We have used CRISPR-Cas9 to knock out the AP-5 ζ subunit gene, AP5Z1, in HeLa cells, and then analysed the phenotype by subcellular fractionation profiling and quantitative mass spectrometry. The retromer complex had an altered steady-state distribution in the knockout cells, and several Golgi proteins, including GOLIM4 and GOLM1, were depleted from vesicle-enriched fractions. Immunolocalisation showed that loss of AP-5 led to impaired retrieval of the cation-independent mannose 6-phosphate receptor (CIMPR, GOLIM4, and GOLM1 from endosomes back to the Golgi region. Knocking down the retromer complex exacerbated this phenotype. Both the CIMPR and sortilin interacted with the AP-5-associated protein SPG15 in pull-down assays, and we propose that sortilin may act as a link between Golgi proteins and the AP-5/SPG11/SPG15 complex. Together, our findings suggest that AP-5 functions in a novel sorting step out of late endosomes, acting as a backup pathway for retromer. This provides a mechanistic explanation for why mutations in AP-5/SPG11/SPG15 cause cells to accumulate aberrant endolysosomes, and highlights the role of endosome/lysosome dysfunction in the pathology of hereditary spastic paraplegia and other neurodegenerative disorders.

  7. Cholesterol Regulates Syntaxin 6 Trafficking at trans-Golgi Network Endosomal Boundaries

    Directory of Open Access Journals (Sweden)

    Meritxell Reverter

    2014-05-01

    Full Text Available Inhibition of cholesterol export from late endosomes causes cellular cholesterol imbalance, including cholesterol depletion in the trans-Golgi network (TGN. Here, using Chinese hamster ovary (CHO Niemann-Pick type C1 (NPC1 mutant cell lines and human NPC1 mutant fibroblasts, we show that altered cholesterol levels at the TGN/endosome boundaries trigger Syntaxin 6 (Stx6 accumulation into VAMP3, transferrin, and Rab11-positive recycling endosomes (REs. This increases Stx6/VAMP3 interaction and interferes with the recycling of αVβ3 and α5β1 integrins and cell migration, possibly in a Stx6-dependent manner. In NPC1 mutant cells, restoration of cholesterol levels in the TGN, but not inhibition of VAMP3, restores the steady-state localization of Stx6 in the TGN. Furthermore, elevation of RE cholesterol is associated with increased amounts of Stx6 in RE. Hence, the fine-tuning of cholesterol levels at the TGN-RE boundaries together with a subset of cholesterol-sensitive SNARE proteins may play a regulatory role in cell migration and invasion.

  8. Augmenting the Efficacy of Immunotoxins and Other Targeted Protein Toxins by Endosomal Escape Enhancers

    Directory of Open Access Journals (Sweden)

    Hendrik Fuchs

    2016-07-01

    Full Text Available The toxic moiety of almost all protein-based targeted toxins must enter the cytosol of the target cell to mediate its fatal effect. Although more than 500 targeted toxins have been investigated in the past decades, no antibody-targeted protein toxin has been approved for tumor therapeutic applications by the authorities to date. Missing efficacy can be attributed in many cases to insufficient endosomal escape and therefore subsequent lysosomal degradation of the endocytosed toxins. To overcome this drawback, many strategies have been described to weaken the membrane integrity of endosomes. This comprises the use of lysosomotropic amines, carboxylic ionophores, calcium channel antagonists, various cell-penetrating peptides of viral, bacterial, plant, animal, human and synthetic origin, other organic molecules and light-induced techniques. Although the efficacy of the targeted toxins was typically augmented in cell culture hundred or thousand fold, in exceptional cases more than million fold, the combination of several substances harbors new problems including additional side effects, loss of target specificity, difficulties to determine the therapeutic window and cell type-dependent variations. This review critically scrutinizes the chances and challenges of endosomal escape enhancers and their potential role in future developments.

  9. Methods of analysis of the membrane trafficking pathway from recycling endosomes to lysosomes.

    Science.gov (United States)

    Matsui, Takahide; Fukuda, Mitsunori

    2014-01-01

    The transferrin receptor (TfR) is responsible for iron uptake through its trafficking between the plasma membrane and recycling endosomes, and as a result it has become a well-known marker for recycling endosomes. Although the molecular basis of the TfR recycling pathway has been thoroughly investigated, the TfR degradation mechanism has been poorly understood. Exposure of cultured cells to two drugs, the protein synthesis inhibitor cycloheximide and the V-ATPase inhibitor bafilomycin A1, recently showed that TfR is not only recycled back to the plasma membrane after endocytosis but is constitutively transported to lysosomes for degradation. The results of genome-wide screening of mouse Rab small GTPases (common regulators of membrane trafficking in all eukaryotes) have indicated that Rab12 regulates TfR trafficking to lysosomes independently of the known membrane trafficking pathways, for example, the conventional endocytic pathway and recycling pathway. This chapter summarizes the methods that the authors used to analyze the membrane trafficking pathway from recycling endosomes to lysosomes that is specifically regulated by Rab12. © 2014 Elsevier Inc. All rights reserved.

  10. Integration of two RAB5 groups during endosomal transport in plants

    Science.gov (United States)

    Ebine, Kazuo; Choi, Seung-won; Ichinose, Sakura; Uemura, Tomohiro; Nakano, Akihiko

    2018-01-01

    RAB5 is a key regulator of endosomal functions in eukaryotic cells. Plants possess two different RAB5 groups, canonical and plant-unique types, which act via unknown counteracting mechanisms. Here, we identified an effector molecule of the plant-unique RAB5 in Arabidopsis thaliana, ARA6, which we designated PLANT-UNIQUE RAB5 EFFECTOR 2 (PUF2). Preferential colocalization with canonical RAB5 on endosomes and genetic interaction analysis indicated that PUF2 coordinates vacuolar transport with canonical RAB5, although PUF2 was identified as an effector of ARA6. Competitive binding of PUF2 with GTP-bound ARA6 and GDP-bound canonical RAB5, together interacting with the shared activating factor VPS9a, showed that ARA6 negatively regulates canonical RAB5-mediated vacuolar transport by titrating PUF2 and VPS9a. These results suggest a unique and unprecedented function for a RAB effector involving the integration of two RAB groups to orchestrate endosomal trafficking in plant cells. PMID:29749929

  11. Involvement of Gβγ subunits of Gi protein coupled with S1P receptor on multivesicular endosomes in F-actin formation and cargo sorting into exosomes.

    Science.gov (United States)

    Kajimoto, Taketoshi; Mohamed, Nesma Nabil Ibrahim; Badawy, Shaymaa Mohamed Mohamed; Matovelo, Shubi Ambwene; Hirase, Mitsuhiro; Nakamura, Shunsuke; Yoshida, Daisuke; Okada, Taro; Ijuin, Takeshi; Nakamura, Shun-Ichi

    2018-01-05

    Exosomes play a critical role in cell-to-cell communication by delivering cargo molecules to recipient cells. However, the mechanism underlying the generation of the exosomal multivesicular endosome (MVE) is one of the mysteries in the field of endosome research. Although sphingolipid metabolites such as ceramide and sphingosine 1-phosphate (S1P) are known to play important roles in MVE formation and maturation, the detailed molecular mechanisms are still unclear. Here, we show that Rho family GTPases, including Cdc42 and Rac1, are constitutively activated on exosomal MVEs and are regulated by S1P signaling as measured by fluorescence resonance energy transfer (FRET)-based conformational changes. Moreover, we detected S1P signaling-induced filamentous actin (F-actin) formation. A selective inhibitor of Gβγ subunits, M119, strongly inhibited both F-actin formation on MVEs and cargo sorting into exosomal intralumenal vesicles of MVEs, both of which were fully rescued by the simultaneous expression of constitutively active Cdc42 and Rac1. Our results shed light on the mechanism underlying exosomal MVE maturation and inform the understanding of the physiological relevance of continuous activation of the S1P receptor and subsequent downstream G protein signaling to Gβγ subunits/Rho family GTPases-regulated F-actin formation on MVEs for cargo sorting into exosomal intralumenal vesicles. © 2018 by The American Society for Biochemistry and Molecular Biology, Inc.

  12. First-Generation Antipsychotic Haloperidol Alters the Functionality of the Late Endosomal/Lysosomal Compartment in Vitro.

    Science.gov (United States)

    Canfrán-Duque, Alberto; Barrio, Luis C; Lerma, Milagros; de la Peña, Gema; Serna, Jorge; Pastor, Oscar; Lasunción, Miguel A; Busto, Rebeca

    2016-03-18

    First- and second-generation antipsychotics (FGAs and SGAs, respectively), have the ability to inhibit cholesterol biosynthesis and also to interrupt the intracellular cholesterol trafficking, interfering with low-density lipoprotein (LDL)-derived cholesterol egress from late endosomes/lysosomes. In the present work, we examined the effects of FGA haloperidol on the functionality of late endosomes/lysosomes in vitro. In HepG2 hepatocarcinoma cells incubated in the presence of 1,1'-dioctadecyl-3,3,3,3'-tetramethylindocarbocyanineperchlorate (DiI)-LDL, treatment with haloperidol caused the enlargement of organelles positive for late endosome markers lysosome-associated membrane protein 2 (LAMP-2) and LBPA (lysobisphosphatidic acid), which also showed increased content of both free-cholesterol and DiI derived from LDL. This indicates the accumulation of LDL-lipids in the late endosomal/lysosomal compartment caused by haloperidol. In contrast, LDL traffic through early endosomes and the Golgi apparatus appeared to be unaffected by the antipsychotic as the distribution of both early endosome antigen 1 (EEA1) and coatomer subunit β (β-COP) were not perturbed. Notably, treatment with haloperidol significantly increased the lysosomal pH and decreased the activities of lysosomal protease and β-d-galactosidase in a dose-dependent manner. We conclude that the alkalinization of the lysosomes' internal milieu induced by haloperidol affects lysosomal functionality.

  13. First-Generation Antipsychotic Haloperidol Alters the Functionality of the Late Endosomal/Lysosomal Compartment in Vitro

    Directory of Open Access Journals (Sweden)

    Alberto Canfrán-Duque

    2016-03-01

    Full Text Available First- and second-generation antipsychotics (FGAs and SGAs, respectively, have the ability to inhibit cholesterol biosynthesis and also to interrupt the intracellular cholesterol trafficking, interfering with low-density lipoprotein (LDL-derived cholesterol egress from late endosomes/lysosomes. In the present work, we examined the effects of FGA haloperidol on the functionality of late endosomes/lysosomes in vitro. In HepG2 hepatocarcinoma cells incubated in the presence of 1,1′-dioctadecyl-3,3,3,3′-tetramethylindocarbocyanineperchlorate (DiI-LDL, treatment with haloperidol caused the enlargement of organelles positive for late endosome markers lysosome-associated membrane protein 2 (LAMP-2 and LBPA (lysobisphosphatidic acid, which also showed increased content of both free-cholesterol and DiI derived from LDL. This indicates the accumulation of LDL-lipids in the late endosomal/lysosomal compartment caused by haloperidol. In contrast, LDL traffic through early endosomes and the Golgi apparatus appeared to be unaffected by the antipsychotic as the distribution of both early endosome antigen 1 (EEA1 and coatomer subunit β (β-COP were not perturbed. Notably, treatment with haloperidol significantly increased the lysosomal pH and decreased the activities of lysosomal protease and β-d-galactosidase in a dose-dependent manner. We conclude that the alkalinization of the lysosomes’ internal milieu induced by haloperidol affects lysosomal functionality.

  14. Basolateral Endocytic Recycling Requires RAB-10 and AMPH-1 Mediated Recruitment of RAB-5 GAP TBC-2 to Endosomes

    Science.gov (United States)

    Liu, Ou; Grant, Barth D.

    2015-01-01

    The small GTPase RAB-5/Rab5 is a master regulator of the early endosome, required for a myriad of coordinated activities, including the degradation and recycling of internalized cargo. Here we focused on the recycling function of the early endosome and the regulation of RAB-5 by GAP protein TBC-2 in the basolateral C. elegans intestine. We demonstrate that downstream basolateral recycling regulators, GTPase RAB-10/Rab10 and BAR domain protein AMPH-1/Amphiphysin, bind to TBC-2 and help to recruit it to endosomes. In the absence of RAB-10 or AMPH-1 binding to TBC-2, RAB-5 membrane association is abnormally high and recycling cargo is trapped in early endosomes. Furthermore, the loss of TBC-2 or AMPH-1 leads to abnormally high spatial overlap of RAB-5 and RAB-10. Taken together our results indicate that RAB-10 and AMPH-1 mediated down-regulation of RAB-5 is an important step in recycling, required for cargo exit from early endosomes and regulation of early endosome–recycling endosome interactions. PMID:26393361

  15. First-Generation Antipsychotic Haloperidol Alters the Functionality of the Late Endosomal/Lysosomal Compartment in Vitro

    Science.gov (United States)

    Canfrán-Duque, Alberto; Barrio, Luis C.; Lerma, Milagros; de la Peña, Gema; Serna, Jorge; Pastor, Oscar; Lasunción, Miguel A.; Busto, Rebeca

    2016-01-01

    First- and second-generation antipsychotics (FGAs and SGAs, respectively), have the ability to inhibit cholesterol biosynthesis and also to interrupt the intracellular cholesterol trafficking, interfering with low-density lipoprotein (LDL)-derived cholesterol egress from late endosomes/lysosomes. In the present work, we examined the effects of FGA haloperidol on the functionality of late endosomes/lysosomes in vitro. In HepG2 hepatocarcinoma cells incubated in the presence of 1,1′-dioctadecyl-3,3,3,3′-tetramethylindocarbocyanineperchlorate (DiI)-LDL, treatment with haloperidol caused the enlargement of organelles positive for late endosome markers lysosome-associated membrane protein 2 (LAMP-2) and LBPA (lysobisphosphatidic acid), which also showed increased content of both free-cholesterol and DiI derived from LDL. This indicates the accumulation of LDL-lipids in the late endosomal/lysosomal compartment caused by haloperidol. In contrast, LDL traffic through early endosomes and the Golgi apparatus appeared to be unaffected by the antipsychotic as the distribution of both early endosome antigen 1 (EEA1) and coatomer subunit β (β-COP) were not perturbed. Notably, treatment with haloperidol significantly increased the lysosomal pH and decreased the activities of lysosomal protease and β-d-galactosidase in a dose-dependent manner. We conclude that the alkalinization of the lysosomes’ internal milieu induced by haloperidol affects lysosomal functionality. PMID:26999125

  16. Photoactive dye-enhanced tissue ablation for endoscopic laser prostatectomy.

    Science.gov (United States)

    Ahn, Minwoo; Hau, Nguyen Trung; Van Phuc, Nguyen; Oh, Junghwan; Kang, Hyun Wook

    2014-11-01

    Laser light has been widely used as a surgical tool to treat benign prostate hyperplasia (BPH) over 20 years. Recently, application of high laser power up to 200 W was often reported to swiftly remove a large amount of prostatic tissue. The purpose of this study was to validate the feasibility of photoactive dye injection to enhance light absorption and eventually to facilitate tissue vaporization with low laser power. Chicken breast tissue was selected as a target tissue due to minimal optical absorption at the visible wavelength. Four biocompatible photoactive dyes, including amaranth (AR), black dye (BD), hemoglobin powder (HP), and endoscopic marker (EM), were selected and tested in vitro with a customized 532 nm laser system with radiant exposure ranging from 0.9 to 3.9 J/cm(2) . Light absorbance and ablation threshold were measured with UV-Vis spectrometer and Probit analysis, respectively, and compared to feature the function of the injected dyes. Ablation performance with dye-injection was evaluated in light of radiant exposure, dye concentration, and number of injection. Higher light absorption by injected dyes led to lower ablation threshold as well as more efficient tissue removal in the order of AR, BD, HP, and EM. Regardless of the injected dyes, ablation efficiency principally increased with radiant exposure, dye concentration, and number of injection. Among the dyes, AR created the highest ablation rate of 44.2 ± 0.2 µm/pulse due to higher absorbance and lower ablation threshold. High aspect ratios up to 7.1 ± 0.4 entailed saturation behavior in the tissue ablation injected with AR and BD, possibly resulting from plume shielding and increased scattering due to coagulation. Preliminary tests on canine prostate with a hydraulic injection system demonstrated that 80 W with dye injection yielded comparable ablation efficiency to 120 W with no injection, indicating 33% reduced laser power with almost equivalent performance. Due to

  17. Radiofrequency Ablation of Liver Tumors

    Science.gov (United States)

    ... have had a surgical procedure in which the liver bile duct has been connected to a loop of bowel are at much greater risk of developing a liver abscess after ablation. Women should always inform their ...

  18. Atrial fibrillation ablation using a closed irrigation radiofrequency ablation catheter.

    Science.gov (United States)

    Golden, Keith; Mounsey, John Paul; Chung, Eugene; Roomiani, Pahresah; Morse, Michael Andew; Patel, Ankit; Gehi, Anil

    2012-05-01

    Catheter ablation is an effective therapy for symptomatic, medically refractory atrial fibrillation (AF). Open-irrigated radiofrequency (RF) ablation catheters produce transmural lesions at the cost of increased fluid delivery. In vivo models suggest closed-irrigated RF catheters create equivalent lesions, but clinical outcomes are limited. A cohort of 195 sequential patients with symptomatic AF underwent stepwise AF ablation (AFA) using a closed-irrigation ablation catheter. Recurrence of AF was monitored and outcomes were evaluated using Kaplan-Meier survival analysis and Cox proportional hazards models. Mean age was 59.0 years, 74.9% were male, 56.4% of patients were paroxysmal and mean duration of AF was 5.4 years. Patients had multiple comorbidities including hypertension (76.4%), tobacco abuse (42.1%), diabetes (17.4%), and obesity (mean body mass index 30.8). The median follow-up was 55.8 weeks. Overall event-free survival was 73.6% with one ablation and 77.4% after reablation (reablation rate was 8.7%). Median time to recurrence was 26.9 weeks. AF was more likely to recur in patients being treated with antiarrhythmic therapy at the time of last follow-up (recurrence rate 30.3% with antiarrhythmic drugs, 13.2% without antiarrhythmic drugs; hazard ratio [HR] 2.2, 95% confidence interval [CI] 1.1-4.4, P = 0.024) and in those with a history of AF greater than 2 years duration (HR 2.7, 95% CI 1.1-6.9, P = 0.038). Our study represents the largest cohort of patients receiving AFA with closed-irrigation ablation catheters. We demonstrate comparable outcomes to those previously reported in studies of open-irrigation ablation catheters. Given the theoretical benefits of a closed-irrigation system, a large head-to-head comparison using this catheter is warranted. ©2012, The Authors. Journal compilation ©2012 Wiley Periodicals, Inc.

  19. Water spray assisted ultrashort laser pulse ablation

    International Nuclear Information System (INIS)

    Silvennoinen, M.; Kaakkunen, J.J.J.; Paivasaari, K.; Vahimaa, P.

    2013-01-01

    Highlights: ► We show the novel method to use multibeam processing with ultrashort pulses efficiently. ► Sprayed thin water layer on ablation zone enhances ablation rate and quality. ► In some cases this method also enables ablation of the deeper and straighter holes compared to ones made without the water layer. ► Method also makes possible to directly write features without the self-organizing structures. - Abstract: We have studied femtosecond ablation under sprayed thin water film and its influence and benefits compared with ablation in the air atmosphere. These have been studied in case of the hole and the groove ablation using IR femtosecond laser. Water enhances the ablation rate and in some situations it makes possible to ablate the holes with a higher aspect ratio. While ablating the grooves, the water spray allows using the high fluences without the generation of the self-organized structures.

  20. Temporal changes in patient characteristics and prior pharmacotherapy in patients undergoing radiofrequency ablation of atrial fibrillation

    DEFF Research Database (Denmark)

    Karasoy, Deniz; Gislason, Gunnar Hilmar; Hansen, Jim

    2013-01-01

    Trends in patient selection and pharmacotherapy before radiofrequency ablation (RFA) of atrial fibrillation are not well studied. We examined temporal trends in RFA utilization on a nationwide scale in Denmark....

  1. Structure and Function of Vps15 in the Endosomal G Protein Signaling Pathway

    Energy Technology Data Exchange (ETDEWEB)

    Heenan, Erin J.; Vanhooke, Janeen L.; Temple, Brenda R.; Betts, Laurie; Sondek, John E.; Dohlman, Henrik G.; (UNC)

    2009-09-11

    G protein-coupled receptors mediate cellular responses to a wide variety of stimuli, including taste, light, and neurotransmitters. In the yeast Saccharomyces cerevisiae, activation of the pheromone pathway triggers events leading to mating. The view had long been held that the G protein-mediated signal occurs principally at the plasma membrane. Recently, it has been shown that the G protein {alpha} subunit Gpa1 can promote signaling at endosomes and requires two components of the sole phosphatidylinositol-3-kinase in yeast, Vps15 and Vps34. Vps15 contains multiple WD repeats and also binds to Gpa1 preferentially in the GDP-bound state; these observations led us to hypothesize that Vps15 may function as a G protein {beta} subunit at the endosome. Here we show an X-ray crystal structure of the Vps15 WD domain that reveals a seven-bladed propeller resembling that of typical G{beta} subunits. We show further that the WD domain is sufficient to bind Gpa1 as well as to Atg14, a potential G{gamma} protein that exists in a complex with Vps15. The Vps15 kinase domain together with the intermediate domain (linking the kinase and WD domains) also contributes to Gpa1 binding and is necessary for Vps15 to sustain G protein signaling. These findings reveal that the Vps15 G{beta}-like domain serves as a scaffold to assemble Gpa1 and Atg14, whereas the kinase and intermediate domains are required for proper signaling at the endosome.

  2. Structural Studies of Adeno-Associated Virus Serotype 8 Capsid Transitions Associated with Endosomal Trafficking

    Energy Technology Data Exchange (ETDEWEB)

    Nam, Hyun-Joo; Gurda, Brittney L.; McKenna, Robert; Potter, Mark; Byrne, Barry; Salganik, Maxim; Muzyczka, Nicholas; Agbandje-McKenna, Mavis (Florida)

    2012-09-17

    The single-stranded DNA (ssDNA) parvoviruses enter host cells through receptor-mediated endocytosis, and infection depends on processing in the early to late endosome as well as in the lysosome prior to nuclear entry for replication. However, the mechanisms of capsid endosomal processing, including the effects of low pH, are poorly understood. To gain insight into the structural transitions required for this essential step in infection, the crystal structures of empty and green fluorescent protein (GFP) gene-packaged adeno-associated virus serotype 8 (AAV8) have been determined at pH values of 6.0, 5.5, and 4.0 and then at pH 7.5 after incubation at pH 4.0, mimicking the conditions encountered during endocytic trafficking. While the capsid viral protein (VP) topologies of all the structures were similar, significant amino acid side chain conformational rearrangements were observed on (i) the interior surface of the capsid under the icosahedral 3-fold axis near ordered nucleic acid density that was lost concomitant with the conformational change as pH was reduced and (ii) the exterior capsid surface close to the icosahedral 2-fold depression. The 3-fold change is consistent with DNA release from an ordering interaction on the inside surface of the capsid at low pH values and suggests transitions that likely trigger the capsid for genome uncoating. The surface change results in disruption of VP-VP interface interactions and a decrease in buried surface area between VP monomers. This disruption points to capsid destabilization which may (i) release VP1 amino acids for its phospholipase A2 function for endosomal escape and nuclear localization signals for nuclear targeting and (ii) trigger genome uncoating.

  3. Direct binding of retromer to human papillomavirus type 16 minor capsid protein L2 mediates endosome exit during viral infection.

    Directory of Open Access Journals (Sweden)

    Andreea Popa

    2015-02-01

    Full Text Available Trafficking of human papillomaviruses to the Golgi apparatus during virus entry requires retromer, an endosomal coat protein complex that mediates the vesicular transport of cellular transmembrane proteins from the endosome to the Golgi apparatus or the plasma membrane. Here we show that the HPV16 L2 minor capsid protein is a retromer cargo, even though L2 is not a transmembrane protein. We show that direct binding of retromer to a conserved sequence in the carboxy-terminus of L2 is required for exit of L2 from the early endosome and delivery to the trans-Golgi network during virus entry. This binding site is different from known retromer binding motifs and can be replaced by a sorting signal from a cellular retromer cargo. Thus, HPV16 is an unconventional particulate retromer cargo, and retromer binding initiates retrograde transport of viral components from the endosome to the trans-Golgi network during virus entry. We propose that the carboxy-terminal segment of L2 protein protrudes through the endosomal membrane and is accessed by retromer in the cytoplasm.

  4. A fluorescence resonance energy transfer-based approach for investigating late endosome-lysosome retrograde fusion events.

    Science.gov (United States)

    Kaufmann, A M; Goldman, S D B; Krise, J P

    2009-03-01

    Traditionally, lysosomes have been considered to be a terminal endocytic compartment. Recent studies suggest that lysosomes are quite dynamic, being able to fuse with other late endocytic compartments as well as with the plasma membrane. Here we describe a quantitative fluorescence energy transfer (FRET)-based method for assessing rates of retrograde fusion between terminal lysosomes and late endosomes in living cells. Late endosomes were specifically labeled with 800-nm latex beads that were conjugated with streptavidin and Alexa Fluor 555 (FRET donor). Terminal lysosomes were specifically labeled with 10,000-MW dextran polymers conjugated with biotin and Alexa Fluor 647 (FRET acceptor). Following late endosome-lysosome fusion, the strong binding affinity between streptavidin and biotin brought the donor and acceptor fluorophore molecules into close proximity, thereby facilitating the appearance of a FRET emission signal. Because apparent size restrictions in the endocytic pathway do not permit endocytosed latex beads from reaching terminal lysosomes in an anterograde fashion, the appearance of the FRET signal is consistent with retrograde transport of lysosomal cargo back to late endosomes. We assessed the efficiency of this transport step in fibroblasts affected by different lysosome storage disorders-Niemann-Pick type C, mucolipidosis type IV, and Sandhoff's disease, all of which have a similar lysosomal lipid accumulation phenotype. We report here, for the first time, that these disorders can be distinguished by their rate of transfer of lysosome cargos to late endosomes, and we discuss the implications of these findings for developing new therapeutic strategies.

  5. Ablation of Solid Hydrogen in a Plasma

    DEFF Research Database (Denmark)

    Jørgensen, L. W.; Sillesen, Alfred Hegaard

    1979-01-01

    Several hydrogen pellet ablation models based on the formation of a shielding neutral cloud have been reported by different authors. The predicted ablation rates are shown to follow almost the same scaling law and this is used to explain the authors' ablation experiment.......Several hydrogen pellet ablation models based on the formation of a shielding neutral cloud have been reported by different authors. The predicted ablation rates are shown to follow almost the same scaling law and this is used to explain the authors' ablation experiment....

  6. Femtosecond laser ablation of enamel

    Science.gov (United States)

    Le, Quang-Tri; Bertrand, Caroline; Vilar, Rui

    2016-06-01

    The surface topographical, compositional, and structural modifications induced in human enamel by femtosecond laser ablation is studied. The laser treatments were performed using a Yb:KYW chirped-pulse-regenerative amplification laser system (560 fs and 1030 nm) and fluences up to 14 J/cm2. The ablation surfaces were studied by scanning electron microscopy, grazing incidence x-ray diffraction, and micro-Raman spectroscopy. Regardless of the fluence, the ablation surfaces were covered by a layer of resolidified material, indicating that ablation is accompanied by melting of hydroxyapatite. This layer presented pores and exploded gas bubbles, created by the release of gaseous decomposition products of hydroxyapatite (CO2 and H2O) within the liquid phase. In the specimen treated with 1-kHz repetition frequency and 14 J/cm2, thickness of the resolidified material is in the range of 300 to 900 nm. The micro-Raman analysis revealed that the resolidified material contains amorphous calcium phosphate, while grazing incidence x-ray diffraction analysis allowed detecting traces of a calcium phosphate other than hydroxyapatite, probably β-tricalcium phosphate Ca3), at the surface of this specimen. The present results show that the ablation of enamel involves melting of enamel's hydroxyapatite, but the thickness of the altered layer is very small and thermal damage of the remaining material is negligible.

  7. A novel chimeric cell-penetrating peptide with membrane-disruptive properties for efficient endosomal escape.

    Science.gov (United States)

    Salomone, Fabrizio; Cardarelli, Francesco; Di Luca, Mariagrazia; Boccardi, Claudia; Nifosì, Riccardo; Bardi, Giuseppe; Di Bari, Lorenzo; Serresi, Michela; Beltram, Fabio

    2012-11-10

    Efficient endocytosis into a wide range of target cells and low toxicity make the arginine-rich Tat peptide (Tat(11): YGRKKRRQRRR, residues 47-57 of HIV-1 Tat protein) an excellent transporter for delivery purposes. Unfortunately, molecules taken up by endocytosis undergo endosomal entrapment and possible metabolic degradation. Escape from the endosome is therefore actively researched. In this context, antimicrobial peptides (AMPs) provide viable templates for the design of new membrane-disruptive motifs. In particular the Cecropin-A and Melittin hybrids (CMs) are among the smallest and most effective peptides with membrane-perturbing abilities. Here we present a novel chimeric peptide in which the Tat(11) motif is fused to the CM(18) hybrid (KWKLFKKIGAVLKVLTTG, residues 1-7 of Cecropin-A and 2-12 of Melittin). When administered to cells, CM(18)-Tat(11) combines the two desired functionalities: efficient uptake and destabilization of endocytotic-vesicle membranes. We show that this chimeric peptide effectively increases cargo-molecule cytoplasm availability and allows the subsequent intracellular localization of diverse membrane-impermeable molecules (i.e. Tat(11)-EGFP fusion protein, calcein, dextrans, and plasmidic DNA) with no detectable cytotoxicity. The present results open the way to the rational engineering of "modular" cell-penetrating peptides (CPPs) that combine (i) efficient translocation from the extracellular milieu into vesicles and (ii) efficient release of molecules from vesicles into the cytoplasm. Copyright © 2012 Elsevier B.V. All rights reserved.

  8. An Inside Job: How Endosomal Na+/H+ Exchangers Link to Autism and Neurological Disease

    Directory of Open Access Journals (Sweden)

    Kalyan C. Kondapalli

    2014-06-01

    Full Text Available Autism imposes a major impediment to childhood development and a huge emotional and financial burden on society. In recent years, there has been rapidly accumulating genetic evidence that links the eNHE, a subset of Na+/H+ exchangers that localize to intracellular vesicles, to a variety of neurological conditions including autism, attention deficit hyperactivity disorder, intellectual disability and epilepsy. By providing a leak pathway for protons pumped by the V-ATPase, eNHE determine luminal pH and regulate cation (Na+, K+ content in early and recycling endosomal compartments. Loss-of-function mutations in eNHE cause hyperacidification of endosomal lumen, as a result of imbalance in pump and leak pathways. Two isoforms, NHE6 and NHE9 are highly expressed in brain, including hippocampus and cortex. Here, we summarize evidence for the importance of luminal cation content and pH on processing, delivery and fate of cargo and on the surface expression and function of membrane receptors and neurotransmitter transporters, drawing upon insights from model organisms and mammalian cells. These studies lead to cellular models of eNHE activity in pre- and post-synaptic neurons and astrocytes, where they could impact synapse development and plasticity. The study of eNHE has provided new insight on the mechanism of autism and other debilitating neurological disorders and opened up new possibilities for therapeutic intervention.

  9. The Annexin A1 Receptor FPR2 Regulates the Endosomal Export of Influenza Virus

    Directory of Open Access Journals (Sweden)

    Fryad Rahman

    2018-05-01

    Full Text Available The Formyl Peptide Receptor 2 (FPR2 is a novel promising target for the treatment of influenza. During viral infection, FPR2 is activated by annexinA1, which is present in the envelope of influenza viruses; this activation promotes virus replication. Here, we investigated whether blockage of FPR2 would affect the genome trafficking of influenza virus. We found that, upon infection and cell treatment with the specific FPR2 antagonist WRW4 or the anti-FPR2 monoclonal antibody, FN-1D6-AI, influenza viruses were blocked into endosomes. This effect was independent on the strain and was observed for H1N1 and H3N2 viruses. In addition, blocking FPR2signaling in alveolar lung A549 epithelial cells with the monoclonal anti-FPR2 antibody significantly inhibited virus replication. Altogether, these results show that FPR2signaling interferes with the endosomal trafficking of influenza viruses and provides, for the first time, the proof of concept that monoclonal antibodies directed against FPR2 inhibit virus replication. Antibodies-based therapeutics have emerged as attractive reagents in infectious diseases. Thus, this study suggests that the use of anti-FPR2 antibodies against influenza hold great promise for the future.

  10. Altered neurological function in mice immunized with early endosome antigen 1

    Directory of Open Access Journals (Sweden)

    Fritzler Marvin J

    2004-01-01

    Full Text Available Abstract Background Autoantibodies directed against the 160 kDa endosome protein early endosome antigen 1 (EEA1 are seen in patients with neurological diseases. To determine if antibodies to EEA1 have a neuropathological effect, mice from three major histocompatability haplotype backgrounds (H2q, H2b and H2d were immunized with EEA1 (amino acids 82–1411 that was previously shown to contain the target EEA1 epitopes. The mice were then subjected to five neuro-behavioural tests: grid walking, forelimb strength, open field, reaching and rotarod. Results The immunized SWR/J mice with sustained anti-EEA1 antibodies had significantly reduced forelimb strength than the control non-immune mice of the same strain, and BALB/CJ immune mice demonstrated significantly more forelimb errors on the grid walk test than the control group. Conclusions Antibodies to recombinant EEA1 in mice may mediate neurological deficits that are consistent with clinical features of some humans that spontaneously develop anti-EEA1 autoantibodies.

  11. Ebola Viral Glycoprotein Bound to Its Endosomal Receptor Niemann-Pick C1.

    Science.gov (United States)

    Wang, Han; Shi, Yi; Song, Jian; Qi, Jianxun; Lu, Guangwen; Yan, Jinghua; Gao, George F

    2016-01-14

    Filoviruses, including Ebola and Marburg, cause fatal hemorrhagic fever in humans and primates. Understanding how these viruses enter host cells could help to develop effective therapeutics. An endosomal protein, Niemann-Pick C1 (NPC1), has been identified as a necessary entry receptor for this process, and priming of the viral glycoprotein (GP) to a fusion-competent state is a prerequisite for NPC1 binding. Here, we have determined the crystal structure of the primed GP (GPcl) of Ebola virus bound to domain C of NPC1 (NPC1-C) at a resolution of 2.3 Å. NPC1-C utilizes two protruding loops to engage a hydrophobic cavity on head of GPcl. Upon enzymatic cleavage and NPC1-C binding, conformational change in the GPcl further affects the state of the internal fusion loop, triggering membrane fusion. Our data therefore provide structural insights into filovirus entry in the late endosome and the molecular basis for design of therapeutic inhibitors of viral entry. Copyright © 2016 Elsevier Inc. All rights reserved.

  12. Huntingtin coordinates the dynein-mediated dynamic positioning of endosomes and lysosomes

    Science.gov (United States)

    Caviston, Juliane P.; Zajac, Allison L.; Tokito, Mariko; Holzbaur, Erika L.F.

    2011-01-01

    Huntingtin (Htt) is a membrane-associated scaffolding protein that interacts with microtubule motors as well as actin-associated adaptor molecules. We examined a role for Htt in the dynein-mediated intracellular trafficking of endosomes and lysosomes. In HeLa cells depleted of either Htt or dynein, early, recycling, and late endosomes (LE)/lysosomes all become dispersed. Despite altered organelle localization, kinetic assays indicate only minor defects in intracellular trafficking. Expression of full-length Htt is required to restore organelle localization in Htt-depleted cells, supporting a role for Htt as a scaffold that promotes functional interactions along its length. In dynein-depleted cells, LE/lysosomes accumulate in tight patches near the cortex, apparently enmeshed by cortactin-positive actin filaments; Latrunculin B-treatment disperses these patches. Peripheral LE/lysosomes in dynein-depleted cells no longer colocalize with microtubules. Htt may be required for this off-loading, as the loss of microtubule association is not seen in Htt-depleted cells or in cells depleted of both dynein and Htt. Inhibition of kinesin-1 relocalizes peripheral LE/lysosomes induced by Htt depletion but not by dynein depletion, consistent with their detachment from microtubules upon dynein knockdown. Together, these data support a model of Htt as a facilitator of dynein-mediated trafficking that may regulate the cytoskeletal association of dynamic organelles. PMID:21169558

  13. Early to Late Endosome Trafficking Controls Secretion and Zymogen Activation in Rodent and Human Pancreatic Acinar Cells.

    Science.gov (United States)

    Messenger, Scott W; Thomas, Diana Dh; Cooley, Michelle M; Jones, Elaina K; Falkowski, Michelle A; August, Benjamin K; Fernandez, Luis A; Gorelick, Fred S; Groblewski, Guy E

    2015-11-01

    Pancreatic acinar cells have an expanded apical endosomal system, the physiological and pathophysiological significance of which is still emerging. Phosphatidylinositol-3,5-bisphosphate (PI(3,5)P 2 ) is an essential phospholipid generated by PIKfyve, which phosphorylates phosphatidylinositol-3-phosphate (PI(3)P). PI(3,5)P 2 is necessary for maturation of early endosomes (EE) to late endosomes (LE). Inhibition of EE to LE trafficking enhances anterograde endosomal trafficking and secretion at the plasma membrane by default through a recycling endosome (RE) intermediate. We assessed the effects of modulating PIKfyve activity on apical trafficking and pancreatitis responses in pancreatic acinar cells. Inhibition of EE to LE trafficking was achieved using pharmacological inhibitors of PIKfyve, expression of dominant negative PIKfyve K1877E, or constitutively active Rab5-GTP Q79L. Anterograde endosomal trafficking was manipulated by expression of constitutively active and dominant negative Rab11a mutants. The effects of these agents on secretion, endolysosomal exocytosis of lysosome associated membrane protein (LAMP1), and trypsinogen activation in response to high-dose CCK-8, bile acids and cigarette toxin was determined. PIKfyve inhibition increased basal and stimulated secretion. Adenoviral overexpression of PIKfyve decreased secretion leading to cellular death. Expression of Rab5-GTP Q79L or Rab11a-GTP Q70L enhanced secretion. Conversely, dominant-negative Rab11a-GDP S25N reduced secretion. High-dose CCK inhibited endolysosomal exocytosis that was reversed by PIKfyve inhibition. PIKfyve inhibition blocked intracellular trypsin accumulation and cellular damage responses to high CCK-8, tobacco toxin, and bile salts in both rodent and human acini. These data demonstrate that EE-LE trafficking acutely controls acinar secretion and the intracellular activation of zymogens leading to the pathogenicity of acute pancreatitis.

  14. Radiofrequency ablation of hepatic metastasis: Results of treatment in forty patients

    Directory of Open Access Journals (Sweden)

    Rath G

    2008-01-01

    Full Text Available Aim: To evaluate the local control of hepatic metastasis with radiofrequency ablation treatment. Materials and Methods: We did a retrospective analysis in 40 patients treated with radiofrequency ablation for hepatic metastasis. The tumors ablated included up to two metastatic liver lesions, with primaries in breast, gastrointestinal tract, cervix, etc. Radiofrequency ablation was performed under general anesthesia in all cases, using ultrasound guidance. Radionics Cool-Tip RF System was used to deliver the treatment. Results: The median age of patients treated was 49 years. There were 13 female and 27 male patients. The median tumor size ablated was 1.5 cm (0.75-4.0 cm. A total of 52 radiofrequency ablation cycles were delivered. Successful ablation was achieved in all patients with hepatic metastasis less than 3 cm in size. Pain was the most common complication seen (75%. One patients developed skin burns. At 2-year follow-up 7.5% of patients had locally recurrent disease. Conclusions: Radiofrequency ablation is a minimally invasive treatment modality. It can be useful in a select group of patients with solitary liver metastasis of less than 3 cm size.

  15. Effects of Arsenic Trioxide on Radiofrequency Ablation of VX2 Liver Tumor: Intraarterial versus Intravenous Administration

    International Nuclear Information System (INIS)

    Seong, Nak Jong; Yoon, Chang Jin; Kang, Sung Gwon; Chung, Jin Wook; Kim, Hyo Cheol; Park, Jae Hyung

    2012-01-01

    Arsenic trioxide (As 2 O 3 ) can be used as a possible pharmaceutical alternative that augments radiofrequency (RF) ablation by reducing tumor blood flow. The aim of this study was to assess the effect of intraarterial and intravenous administration of As 2 O 3 on RF-induced ablation in an experimentally induced liver tumor. VX2 carcinoma was grown in the livers of 30 rabbits. As 2 O 3 (1 mg/kg) was administered through the hepatic artery (n = 10, group A) or ear vein (n = 10, group B), 30 minutes before RF ablation (125 mA ± 35; 90 ± 5 degrees Celsius). As a control group, 10 rabbits were treated with RF ablation alone (group C). RF was intentionally applied to the peripheral margin of the tumor so that ablation can cover the tumor and adjacent hepatic parenchyma. Ablation areas of the tumor and adjacent parenchymal changes among three groups were compared by the Kruskal-Wallis and Mann-Whitney U test. The overall ablation areas were 156 ± 28.9 mm 2 (group A), 119 ± 31.7 (group B), and 92 ± 17.4 (group C, p 2 ) than both group B (50 ± 19.4, p = 0.02) and group C (28 ± 2.2, p 2 O 3 . The intraarterial administration of As 2 O 3 seems to be helpful for the selective ablation of the tumor.

  16. The NKG2D ligand ULBP2 is specifically regulated through an invariant chain-dependent endosomal pathway

    DEFF Research Database (Denmark)

    Uhlenbrock, Franziska Katharina; Hagemann-Jensen, Michael Henrik; Kehlet, Stephanie

    2014-01-01

    by affecting endosomal/lysosomal integrity and protein kinase C activity. The invariant chain was further essential for endosomal transport of ULBP2. This novel pathway was identified through screening experiments by which methylselenic acid was found to possess notable NKG2D ligand regulatory properties....... The protein kinase C inhibitor methylselenic acid induced MICA/B surface expression but dominantly blocked ULBP2 surface transport. Remarkably, by targeting this novel pathway we could specifically block the production of soluble ULBP2 from different, primary melanomas. Our findings strongly suggest...

  17. Femtosecond laser ablation of dentin

    International Nuclear Information System (INIS)

    Alves, S; Vilar, R; Oliveira, V

    2012-01-01

    The surface morphology, structure and composition of human dentin treated with a femtosecond infrared laser (pulse duration 500 fs, wavelength 1030 nm, fluences ranging from 1 to 3 J cm -2 ) was studied by scanning electron microscopy, x-ray diffraction, x-ray photoelectron spectroscopy and Fourier transform infrared spectroscopy. The average dentin ablation threshold under these conditions was 0.6 ± 0.2 J cm -2 and the ablation rate achieved in the range 1 to 2 µm/pulse for an average fluence of 3 J cm -2 . The ablation surfaces present an irregular and rugged appearance, with no significant traces of melting, deformation, cracking or carbonization. The smear layer was entirely removed by the laser treatment. For fluences only slightly higher than the ablation threshold the morphology of the laser-treated surfaces was very similar to the dentin fracture surfaces and the dentinal tubules remained open. For higher fluences, the surface was more porous and the dentin structure was partially concealed by ablation debris and a few resolidified droplets. Independently on the laser processing parameters and laser processing method used no sub-superficial cracking was observed. The dentin constitution and chemical composition was not significantly modified by the laser treatment in the processing parameter range used. In particular, the organic matter is not preferentially removed from the surface and no traces of high temperature phosphates, such as the β-tricalcium phosphate, were observed. The achieved results are compatible with an electrostatic ablation mechanism. In conclusion, the high beam quality and short pulse duration of the ultrafast laser used should allow the accurate preparation of cavities, with negligible damage of the underlying material. (paper)

  18. Skin pre-ablation and laser assisted microjet injection for deep tissue penetration.

    Science.gov (United States)

    Jang, Hun-Jae; Yeo, Seonggu; Yoh, Jack J

    2017-04-01

    For conventional needless injection, there still remain many unresolved issues such as the potential for cross-contamination, poor reliability of targeted delivery dose, and significantly painstaking procedures. As an alternative, the use of microjets generated with Er:YAG laser for delivering small doses with controlled penetration depths has been reported. In this study, a new system with two stages is evaluated for effective transdermal drug delivery. First, the skin is pre-ablated to eliminate the hard outer layer and second, laser-driven microjet penetrates the relatively weaker and freshly exposed epidermis. Each stage of operation shares a single Er:YAG laser that is suitable for skin ablation as well as for the generation of a microjet. In this study, pig skin is selected for quantification of the injection depth based on the two-stage procedure, namely pre-ablation and microjet injection. The three types of pre-ablation devised here consists of bulk ablation, fractional ablation, and fractional-rotational ablation. The number of laser pulses are 12, 18, and 24 for each ablation type. For fractional-rotational ablation, the fractional beams are rotated by 11.25° at each pulse. The drug permeation in the skin is evaluated using tissue marking dyes. The depth of penetration is quantified by a cross sectional view of the single spot injections. Multi-spot injections are also carried out to control the dose and spread of the drug. The benefits of a pre-ablation procedure prior to the actual microjet injection to the penetration is verified. The four possible combinations of injection are (a) microjet only; (b) bulk ablation and microjet injection; (c) fractional ablation and microjet injection; and (d) fractional-rotational ablation and microjet injection. Accordingly, the total depth increases with injection time for all cases. In particular, the total depth of penetration attained via fractional pre-ablation increased by 8 ∼ 11% and that of fractional

  19. Transhemangioma Ablation of Hepatocellular Carcinoma

    International Nuclear Information System (INIS)

    Pua, Uei

    2012-01-01

    Radiofrequency ablation (RFA) is a well-established treatment modality in the treatment of early hepatocellular carcinoma (HCC) [1]. Safe trajectory of the RFA probe is crucial in decreasing collateral tissue damage and unwarranted probe transgression. As a percutaneous technique, however, the trajectory of the needle is sometimes constrained by the available imaging plane. The presence of a hemangioma beside an HCC is uncommon but poses the question of safety related to probe transgression. We hereby describe a case of transhemangioma ablation of a dome HCC.

  20. Transhemangioma Ablation of Hepatocellular Carcinoma

    Energy Technology Data Exchange (ETDEWEB)

    Pua, Uei, E-mail: druei@yahoo.com [Tan Tock Seng Hospital, Department of Diagnostic Radiology (Singapore)

    2012-12-15

    Radiofrequency ablation (RFA) is a well-established treatment modality in the treatment of early hepatocellular carcinoma (HCC) [1]. Safe trajectory of the RFA probe is crucial in decreasing collateral tissue damage and unwarranted probe transgression. As a percutaneous technique, however, the trajectory of the needle is sometimes constrained by the available imaging plane. The presence of a hemangioma beside an HCC is uncommon but poses the question of safety related to probe transgression. We hereby describe a case of transhemangioma ablation of a dome HCC.

  1. Selective ablation of a titanium nitride film on tungsten carbide substrate using ultrashort laser pulses; Ablação seletiva de um filme de nitreto de titânio em substrato de carboneto de tungstênio utilizando laser de pulsos ultracurtos

    Energy Technology Data Exchange (ETDEWEB)

    Oliveira, Eduardo Spinelli

    2017-07-01

    Surface coatings are applied to many cutting tools in the metallurgical industry in order to improve cutting efficiency and extend its useful life. In this work, tests were performed to remove the coating of titanium aluminum nitride (TiAlN) on tungsten carbide (WC-Co) pellets, using an ultrashort laser pulses beam. After determination of the damage thresholds of the film and the substrate, were ablated on the surface of the coating lines using two ablation conditions, it was initially operated on the low fluence regime for the film, and later on the low fluence regime of the substrate, far below the threshold of the film, applying high overlapping pulses. A laser induced breakdown spectroscopy (LIBS) system was set up to monitor the materials present in the plasma generated by the laser, but the system did not present sufficient sensitivity to read the low intensity of the plasma generated in the process and was not used. After the analysis of the traces by electron microscopy, optical profilometer and X-ray fluorescence spectroscopy, it was not possible to determine a safe process to carry out the selective removal of the film in question, however, due to the data obtained and observations of the results in some traces, new possibilities were raised, opening the discussion for future work. (author)

  2. The N-terminal domains of Vps3 and Vps8 are critical for localization and function of the CORVET tethering complex on endosomes.

    Directory of Open Access Journals (Sweden)

    Nadine Epp

    Full Text Available Endosomal biogenesis depends on multiple fusion and fission events. For fusion, the heterohexameric CORVET complex as an effector of the endosomal Rab5/Vps21 GTPase has a central function in the initial tethering event. Here, we show that the CORVET-specific Vps3 and Vps8 subunits, which interact with Rab5/Vps21, require their N-terminal domains for localization and function. Surprisingly, CORVET may lack either one of the two N-terminal domains, but not both, to promote protein sorting via the endosome. The dually truncated complex mislocalizes to the cytosol and is impaired in endocytic protein sorting, but not in assembly. Furthermore, the endosomal localization can be rescued by overexpression of Vps21 or one of the truncated CORVET subunits, even though CORVET assembly is not impaired by loss of the N-terminal domains or in strains lacking all endosomal Rab5s and Ypt7. We thus conclude that CORVET requires only its C-terminal domains for assembly and has beyond its putative β-propeller domains additional binding sites for endosomes, which could be important to bind Vps21 and other endosome-specific factors for efficient endosome tethering.

  3. Fractional ablative laser skin resurfacing: a review.

    Science.gov (United States)

    Tajirian, Ani L; Tarijian, Ani L; Goldberg, David J

    2011-12-01

    Ablative laser technology has been in use for many years now. The large side effect profile however has limited its use. Fractional ablative technology is a newer development which combines a lesser side effect profile along with similar efficacy. In this paper we review fractional ablative laser skin resurfacing.

  4. HIV-1 Envelope Glycoprotein Trafficking through the Endosomal Recycling Compartment Is Required for Particle Incorporation.

    Science.gov (United States)

    Kirschman, Junghwa; Qi, Mingli; Ding, Lingmei; Hammonds, Jason; Dienger-Stambaugh, Krista; Wang, Jaang-Jiun; Lapierre, Lynne A; Goldenring, James R; Spearman, Paul

    2018-03-01

    The human immunodeficiency virus type 1 (HIV-1) envelope glycoprotein (Env) encodes specific trafficking signals within its long cytoplasmic tail (CT) that regulate incorporation into HIV-1 particles. Rab11-family interacting protein 1C (FIP1C) and Rab14 are host trafficking factors required for Env particle incorporation, suggesting that Env undergoes sorting from the endosomal recycling compartment (ERC) to the site of particle assembly on the plasma membrane. We disrupted outward sorting from the ERC by expressing a C-terminal fragment of FIP1C (FIP1C 560-649 ) and examined the consequences on Env trafficking and incorporation into particles. FIP1C 560-649 reduced cell surface levels of Env and prevented its incorporation into HIV-1 particles. Remarkably, Env was trapped in an exaggerated perinuclear ERC in a CT-dependent manner. Mutation of either the Yxxϕ endocytic motif or the YW 795 motif in the CT prevented Env trapping in the ERC and restored incorporation into particles. In contrast, simian immunodeficiency virus SIVmac239 Env was not retained in the ERC, while substitution of the HIV-1 CT for the SIV CT resulted in SIV Env retention in this compartment. These results provide the first direct evidence that Env traffics through the ERC and support a model whereby HIV-1 Env is specifically targeted to the ERC prior to FIP1C- and CT-dependent outward sorting to the particle assembly site on the plasma membrane. IMPORTANCE The HIV envelope protein is an essential component of the viral particle. While many aspects of envelope protein structure and function have been established, the pathway it follows in the cell prior to reaching the site of particle assembly is not well understood. The envelope protein has a very long cytoplasmic tail that interacts with the host cell trafficking machinery. Here, we utilized a truncated form of the trafficking adaptor FIP1C protein to arrest the intracellular transport of the envelope protein, demonstrating that it becomes

  5. Endometrial ablation with paracervical block

    NARCIS (Netherlands)

    Penninx, Josien P. M.; Mol, Ben Willem; Bongers, Marlies Y.

    2009-01-01

    OBJECTIVE: To evaluate the safety, feasibility and efficacy of endometrial ablation under local anesthesia. STUDY DESIGN: A prospective cohort study was performed at the gynecology department of a large teaching hospital. Women with dysfunctional uterine bleeding were included to undergo NovaSure

  6. Mucosal ablation in Barrett's esophagus.

    Science.gov (United States)

    Walker, S J; Selvasekar, C R; Birbeck, N

    2002-01-01

    Barrett's esophagus is a prevalent, premalignant condition affecting the gastroesophageal junction and distal esophagus. Ablation plus antireflux therapy has recently been advocated to prevent the development of adenocarcinoma or to treat those unfit or unwilling to undergo esophagectomy. The present article, based on a search of Medline/ISI databases and cross-referencing of relevant articles, reviews the literature on this subject. A number of techniques have been used to remove the affected mucosa, including laser, electrocoagulation, argon plasma coagulation and photodynamic therapy but, as yet, none has been shown to be superior. Depending on the method used, ablation results in complete removal of Barrett's esophagus in approximately one third of patients and a partial response in nearly two-thirds. The resultant squamous mucosa is apparently 'normal' but may regress. To promote and maintain regeneration, antireflux therapy must be sufficient to reduce repetitive injury to the esophageal mucosa. Whether ablation reduces the cancer risk or delays its occurrence is unknown, though recent data suggests benefit. Complications are infrequent and usually mild. Regular follow-up endoscopy and deep biopsies continue to be necessary. Careful data from much larger populations with long-term follow-up is required before ablation reaches the stage of broad clinical application.

  7. Hydrodynamic instabilities in an ablation front

    International Nuclear Information System (INIS)

    Piriz, A R; Portugues, R F

    2004-01-01

    The hydrodynamic stability of an ablation front is studied for situations in which the wavelength of the perturbations is larger than the distance to the critical surface where the driving radiation is absorbed. An analytical model is presented, and it shows that under conditions in which the thermal flux is limited within the supercritical region of the ablative corona, the front may behave like a flame or like an ablation front, depending on the perturbation wavelength. For relatively long wavelengths the critical and ablation surfaces practically lump together into a unique surface and the front behaves like a flame, whereas for the shortest wavelengths the ablation front substructure is resolved

  8. Hydrodynamic instabilities in an ablation front

    Energy Technology Data Exchange (ETDEWEB)

    Piriz, A R; Portugues, R F [E.T.S.I. Industriales, Universidad de Castilla-La Mancha, 13071 Ciudad Real (Spain)

    2004-06-01

    The hydrodynamic stability of an ablation front is studied for situations in which the wavelength of the perturbations is larger than the distance to the critical surface where the driving radiation is absorbed. An analytical model is presented, and it shows that under conditions in which the thermal flux is limited within the supercritical region of the ablative corona, the front may behave like a flame or like an ablation front, depending on the perturbation wavelength. For relatively long wavelengths the critical and ablation surfaces practically lump together into a unique surface and the front behaves like a flame, whereas for the shortest wavelengths the ablation front substructure is resolved.

  9. Comparison of wet radiofrequency ablation with dry radiofrequency ablation and radiofrequency ablation using hypertonic saline preinjection: ex vivo bovine liver

    International Nuclear Information System (INIS)

    Lee, Jeong Min; Han, Joon Koo; Kim, Se Hyung; Lee, Jae Young; Park, Hee Sun; Hur, Hurn; Choi, Byung Ihn; Shin, Kyung Sook

    2004-01-01

    We wished to compare the in-vitro efficiency of wet radiofrequency (RF) ablation with the efficiency of dry RF ablation and RF ablation with preinjection of NaCl solutions using excised bovine liver. Radiofrequency was applied to excised bovine livers in a monopolar mode for 10 minutes using a 200 W generator and a perfused-cooled electrode with or without injection or slow infusion of NaCl solutions. After placing the perfused-cooled electrode in the explanted liver, 50 ablation zones were created with five different regimens: group A; standard dry RF ablation, group B; RF ablation with 11 mL of 5% NaCl solution preinjection, group C; RF ablation with infusion of 11 mL of 5% NaCl solution at a rate of 1 mL/min, group D; RFA with 6 mL of 36% NaCl solution preinjection, group E; RF ablation with infusion of 6 mL of 36% NaCl solution at a rate of 0.5 mL/min. In groups C and E, infusion of the NaCl solutions was started 1 min before RF ablation and then maintained during RF ablation (wet RF ablation). During RF ablation, we measured the tissue temperature at 15 mm from the electrode. The dimensions of the ablation zones and changes in impedance, current and liver temperature during RF ablation were then compared between the groups. With injection or infusion of NaCl solutions, the mean initial tissue impedance prior to RF ablation was significantly less in groups B, C, D, and E (43-75 Ω) than for group A (80 Ω) (ρ 3 in group A; 12.4 ± 3.8 cm 3 in group B; 80.9 ± 9.9 cm 3 in group C; 45.3 ± 11.3 cm 3 in group D and 81.6 ± 8.6 cm 3 in group E. The tissue temperature measured at 15 mm from the electrode was higher in groups C, D and E than other groups (ρ < 0.05): 53 ± 12 .deg. C in group A, 42 ± 2 .deg. C in group B, 93 ± 8 .deg. C in group C; 79 ± 12 .deg. C in group D and 83 ± 8 .deg.C in group E. Wet RF ablation with 5% or 36% NaCl solutions shows better efficiency in creating a large ablation zone than does dry RF ablation or RF ablation with

  10. Study of radiative ablation to low-Z material and energy transport

    International Nuclear Information System (INIS)

    Yang Jiamin; Ding Yaonan; Miao Wenyong; Sun Kexu; Yi Rongqing; Chen Zhenglin; Wang Hongbin; Li Sanwei; Wang Yaomei; Wen Shuhuai; Zheng Zhijian; Zhang Wenhai; Yu Yanning

    1998-12-01

    X-ray emissions from the gold foil target, irradiated by 0.35 μm laser on the Xingguang facility, have been studied. A clean and intense X-ray source has been obtained from the rear of gold foil target by selection of irradiating laser parameters. Then, characteristics of radiation ablation to low-Z materials C 8 H 8 and C 10 H 16 O 5 and energy transport have been investigated comprehensively. Experimental results show that mass ablative rate of C 10 H 16 O 5 are greater than those of C 8 H 8 due to its better match with the ablative source spectra

  11. The contemporary role of ablative treatment approaches in the management of renal cell carcinoma (RCC): focus on radiofrequency ablation (RFA), high-intensity focused ultrasound (HIFU), and cryoablation.

    Science.gov (United States)

    Klatte, Tobias; Kroeger, Nils; Zimmermann, Uwe; Burchardt, Martin; Belldegrun, Arie S; Pantuck, Allan J

    2014-06-01

    Currently, most of renal tumors are small, low grade, with a slow growth rate, a low metastatic potential, and with up to 30 % of these tumors being benign on the final pathology. Moreover, they are often diagnosed in elderly patients with preexisting medical comorbidities in whom the underlying medical conditions may pose a greater risk of death than the small renal mass. Concerns regarding overdiagnosis and overtreatment of patients with indolent small renal tumors have led to an increasing interest in minimally invasive, ablative as an alternative to extirpative interventions for selected patients. To provide an overview about the state of the art in radiofrequency ablation (RFA), high-intensity focused ultrasound, and cryoablation in the clinical management of renal cell carcinoma. A PubMed wide the literature search of was conducted. International consensus panels recommend ablative techniques in patients who are unfit for surgery, who are not considered candidates for or elect against elective surveillance, and who have small renal masses. The most often used techniques are cryoablation and RFA. These ablative techniques offer potentially curative outcomes while conferring several advantages over extirpative surgery, including improved patient procedural tolerance, faster recovery, preservation of renal function, and reduction in the risk of intraoperative and postsurgical complications. While it is likely that outcomes associated with ablative modalities will improve with further advances in technology, their application will expand to more elective indications as longer-term efficacy data become available. Ablative techniques pose a valid treatment option in selected patients.

  12. Radiofrequency ablation of liver metastases; Radiofrequenzablation von Lebermetastasen

    Energy Technology Data Exchange (ETDEWEB)

    Pereira, P.L.; Clasen, S.; Schmidt, D.; Wiskirchen, J.; Tepe, G.; Claussen, C.D. [Abt. fuer Radiologische Diagnostik, Eberhard-Karls-Univ. Tuebingen (Germany); Boss, A. [Abt. fuer Radiologische Diagnostik, Eberhard-Karls-Univ. Tuebingen (Germany); Sektion fuer Experimentelle Radiologie der Abt. fuer Radiologische Diagnostik, Eberhard-Karls-Univ. Tuebingen (Germany); Gouttefangeas, C. [Abt. Immunologie des Inst. fuer Zellbiologie, Eberhard-Karls-Univ. Tuebingen (Germany); Burkart, C. [Zentrum fuer gastroenterologische Onkologie der Medizinischen Klinik, Eberhard-Karls-Univ. Tuebingen (Germany)

    2004-04-01

    The liver is the second only to lymph nodes as the most common site of metastatic disease irrespective of the primary tumor. Up to 50% of all patients with malignant diseases will develop liver metastases with a significant morbidity and mortality. Although the surgical resection leads to an improvement of the survival time, only approximately 20% of the patients are eligible for surgical intervention. Radiofrequency (RF) ablation represents one of the most important alternatives as well as complementary methods for the therapy of liver metastases. RF ablation can lead in a selected patient group to a palliation or to an increased life expectancy. RF ablation appears either safer (vs. cryotherapy) or easier (vs. laser) or more effective (percutaneous ethanol instillation [PEI]), transarterial chemoembolisation [TACE] in comparison with other minimal invasive procedures. RF ablation can be performed percutaneously, laparoscopically or intraoperatively and may be combined with chemotherapy as well as with surgical resection. Permanent technical improvements of RF systems, a better understanding of the underlying electrophysiological principles and an interdisciplinary approach will lead to a prognosis improvement in patients with liver metastases. (orig.) [German] Die Leber ist unabhaengig vom Primaertumor nach den Lymphknoten die zweithaeufigste Lokalisation von Metastasen. Bis zu 50% aller Patienten mit malignen Erkrankungen werden im Verlauf ihrer Erkrankung Lebermetastasen entwickeln, die mit einer signifikanten Morbiditaet und Mortalitaet verbunden sind. Obwohl die chirurgische Resektion zu einer verlaengerten Ueberlebenszeit fuehrt, sind nur ca. 20% der Patienten fuer einen chirurgischen Eingriff geeignet. Die Radiofrequenz-(RF-)Ablation stellt derzeit eine der effektivsten Alternativen und komplementaeren Methoden bei der Therapie von Lebermetastasen dar. In einem selektierten Patientengut fuehrt die RF-Ablation ueber den palliativen Einsatz hinaus zu einer

  13. Kinesin-3 and dynein cooperate in long-range retrograde endosome motility along a nonuniform microtubule array

    NARCIS (Netherlands)

    Schuster, M.; Kilaru, S.; Fink, G.; Collemare, J.A.R.; Roger, Y.; Steinberg, G.

    2011-01-01

    The polarity of microtubules (MTs) determines the motors for intracellular motility, with kinesins moving to plus ends and dynein to minus ends. In elongated cells of Ustilago maydis, dynein is thought to move early endosomes (EEs) toward the septum (retrograde), whereas kinesin-3 transports them to

  14. Characterization of PEBBLEs as a Tool for Real-Time Measurement of Dictyostelium discoideum Endosomal pH

    Directory of Open Access Journals (Sweden)

    Everett Moding

    2009-01-01

    Full Text Available The measurement of intracellular ion concentration change is important for understanding the cellular mechanisms for communication. Recently developed nanosensors, (Photonic Explorers for Biomedical use with Biologically Localized Embedding PEBBLEs, have a number of advantages for measuring ions in cells over established methods using microelectrodes, unbound fluorescent dyes, or NMR. PEBBLE sensors have been shown to work in principle for measuring dynamic ion changes, but few in vivo applications have been demonstrated. We modified the protocol for the fabrication of pH sensing PEBBLEs and developed a protocol for the utilization of these sensors for the monitoring of dynamic pH changes in the endosomes of slime mold Dictyostelium discoideum (D. discoideum. Oregon Green 514-CdSe Quantum Dot PEBBLEs were used to measure real-time pH inside D. discoideum endosomes during cAMP stimulation. Endosomal pH was shown to decrease during cAMP signaling, demonstrating a movement of protons into the endosomes of D. discoideum amoebae.

  15. COMMD1 is linked to the WASH complex and regulates endosomal trafficking of the copper transporter ATP7A

    NARCIS (Netherlands)

    Phillips-Krawczak, Christine A.; Singla, Amika; Starokadomskyy, Petro; Deng, Zhihui; Osborne, Douglas G.; Li, Haiying; Dick, Christopher J.; Gomez, Timothy S.; Koenecke, Megan; Zhang, Jin-San; Dai, Haiming; Sifuentes-Dominguez, Luis F.; Geng, Linda N.; Kaufmann, Scott H.; Hein, Marco Y.; Wallis, Mathew; McGaughran, Julie; Gecz, Jozef; De Sluis, Bart van; Billadeau, Daniel D.; Burstein, Ezra

    2015-01-01

    COMMD1 deficiency results in defective copper homeostasis, but the mechanism for this has remained elusive. Here we report that COMMD1 is directly linked to early endosomes through its interaction with a protein complex containing CCDC22, CCDC93, and C16orf62. This COMMD/CCDC22/CCDC93 (CCC) complex

  16. Localization of the AP-3 adaptor complex defines a novel endosomal exit site for lysosomal membrane proteins

    NARCIS (Netherlands)

    Peden, A.A.; Oorschot, V.; Hesser, B.A.; Austin, C.D.; Scheller, R.H.; Klumperman, J.

    2004-01-01

    The adaptor protein (AP) 3 adaptor complex has been implicated in the transport of lysosomal membrane proteins, but its precise site of action has remained controversial. Here, we show by immuno-electron microscopy that AP-3 is associated with budding profiles evolving from a tubular endosomal

  17. CCC- and WASH-mediated endosomal sorting of LDLR is required for normal clearance of circulating LDL

    NARCIS (Netherlands)

    Bartuzi, Paulina; Billadeau, Daniel D.; Favier, Robert; Rong, Shunxing; Dekker, Daphne; Fedoseienko, Alina; Fieten, Hille; Wijers, Melinde; Levels, Johannes H.; Huijkman, Nicolette; Kloosterhuis, Niels; Van der Molen, Henk; Brufau, Gemma; Groen, Albert K.; Elliott, Alison M.; Kuivenhoven, Jan Albert; Plecko, Barbara; Grangl, Gernot; McGaughran, Julie; Horton, Jay D.; Burstein, Ezra; Hofker, Marten H.; van de Sluis, Bart

    2016-01-01

    The low-density lipoprotein receptor (LDLR) plays a pivotal role in clearing atherogenic circulating low-density lipoprotein (LDL) cholesterol. Here we show that the COMMD/CCDC22/CCDC93 (CCC) and the Wiskott-Aldrich syndrome protein and SCAR homologue (WASH) complexes are both crucial for endosomal

  18. CCC- and WASH-mediated endosomal sorting of LDLR is required for normal clearance of circulating LDL

    NARCIS (Netherlands)

    Bartuzi, Paulina; Billadeau, Daniel D.; Favier, Robert; Rong, Shunxing; Dekker, Daphne; Fedoseienko, Alina; Fieten, Hille; Wijers, Melinde; Levels, Johannes H.; Huijkman, Nicolette; Kloosterhuis, Niels; van der Molen, Henk; Brufau, Gemma; Groen, Albert K.; Elliott, Alison M.; Kuivenhoven, Jan Albert; Plecko, Barbara; Grangl, Gernot; McGaughran, Julie; Horton, Jay D.; Burstein, Ezra; Hofker, Marten H.; van de Sluis, Bart

    The low-density lipoprotein receptor (LDLR) plays a pivotal role in clearing atherogenic circulating low-density lipoprotein (LDL) cholesterol. Here we show that the COMMD/CCDC22/CCDC93 (CCC) and the Wiskott-Aldrich syndrome protein and SCAR homologue (WASH) complexes are both crucial for endosomal

  19. Kinesin Khc-73/KIF13B modulates retrograde BMP signaling by influencing endosomal dynamics at the Drosophila neuromuscular junction.

    Science.gov (United States)

    Liao, Edward H; Gray, Lindsay; Tsurudome, Kazuya; El-Mounzer, Wassim; Elazzouzi, Fatima; Baim, Christopher; Farzin, Sarah; Calderon, Mario R; Kauwe, Grant; Haghighi, A Pejmun

    2018-01-01

    Retrograde signaling is essential for neuronal growth, function and survival; however, we know little about how signaling endosomes might be directed from synaptic terminals onto retrograde axonal pathways. We have identified Khc-73, a plus-end directed microtubule motor protein, as a regulator of sorting of endosomes in Drosophila larval motor neurons. The number of synaptic boutons and the amount of neurotransmitter release at the Khc-73 mutant larval neuromuscular junction (NMJ) are normal, but we find a significant decrease in the number of presynaptic release sites. This defect in Khc-73 mutant larvae can be genetically enhanced by a partial genetic loss of Bone Morphogenic Protein (BMP) signaling or suppressed by activation of BMP signaling in motoneurons. Consistently, activation of BMP signaling that normally enhances the accumulation of phosphorylated form of BMP transcription factor Mad in the nuclei, can be suppressed by genetic removal of Khc-73. Using a number of assays including live imaging in larval motor neurons, we show that loss of Khc-73 curbs the ability of retrograde-bound endosomes to leave the synaptic area and join the retrograde axonal pathway. Our findings identify Khc-73 as a regulator of endosomal traffic at the synapse and modulator of retrograde BMP signaling in motoneurons.

  20. IL4/PGE2 induction of an enlarged early endosomal compartment in mouse macrophages is Rab5-dependent

    International Nuclear Information System (INIS)

    Wainszelbaum, Marisa J.; Proctor, Brandon M.; Pontow, Suzanne E.; Stahl, Philip D.; Barbieri, M. Alejandro

    2006-01-01

    The endosomal compartment and the plasma membrane form a complex partnership that controls signal transduction and trafficking of different molecules. The specificity and functionality of the early endocytic pathway are regulated by a growing number of Rab GTPases, particularly Rab5. In this study, we demonstrate that IL4 (a Th-2 cytokine) and prostaglandin E 2 (PGE 2 ) synergistically induce Rab5 and several Rab effector proteins, including Rin1 and EEA1, and promote the formation of an enlarged early endocytic (EEE) compartment. Endosome enlargement is linked to a substantial induction of the mannose receptor (MR), a well-characterized macrophage endocytic receptor. Both MR levels and MR-mediated endocytosis are enhanced approximately 7-fold. Fluid-phase endocytosis is also elevated in treated cells. Light microscopy and fractionation studies reveal that MR colocalizes predominantly with Rab5a and partially with Rab11, an endosomal recycling pathway marker. Using retroviral expression of Rab5a:S34N, a dominant negative mutant, and siRNA Rab5a silencing, we demonstrate that Rab5a is essential for the large endosome phenotype and for localization of MR in these structures. We speculate that the EEE is maintained by activated Rab5, and that the EEE phenotype is part of some macrophage developmental program such as cell fusion, a characteristic of IL4-stimulated cells

  1. Transport, metabolism, and endosomal trafficking-dependent regulation of intestinal fructose absorption

    Science.gov (United States)

    Patel, Chirag; Douard, Veronique; Yu, Shiyan; Gao, Nan; Ferraris, Ronaldo P.

    2015-01-01

    Dietary fructose that is linked to metabolic abnormalities can up-regulate its own absorption, but the underlying regulatory mechanisms are not known. We hypothesized that glucose transporter (GLUT) protein, member 5 (GLUT5) is the primary fructose transporter and that fructose absorption via GLUT5, metabolism via ketohexokinase (KHK), as well as GLUT5 trafficking to the apical membrane via the Ras-related protein-in-brain 11 (Rab11)a-dependent endosomes are each required for regulation. Introducing fructose but not lysine and glucose solutions into the lumen increased by 2- to 10-fold the heterogeneous nuclear RNA, mRNA, protein, and activity levels of GLUT5 in adult wild-type mice consuming chow. Levels of GLUT5 were >100-fold that of candidate apical fructose transporters GLUTs 7, 8, and 12 whose expression, and that of GLUT 2 and the sodium-dependent glucose transporter protein 1 (SGLT1), was not regulated by luminal fructose. GLUT5-knockout (KO) mice exhibited no facilitative fructose transport and no compensatory increases in activity and expression of SGLT1 and other GLUTs. Fructose could not up-regulate GLUT5 in GLUT5-KO, KHK-KO, and intestinal epithelial cell-specific Rab11a-KO mice. The fructose-specific metabolite glyceraldehyde did not increase GLUT5 expression. GLUT5 is the primary transporter responsible for facilitative absorption of fructose, and its regulation specifically requires fructose uptake and metabolism and normal GLUT5 trafficking to the apical membrane.—Patel, C., Douard, V., Yu, S., Gao, N., Ferraris, R. P. Transport, metabolism, and endosomal trafficking-dependent regulation of intestinal fructose absorption. PMID:26071406

  2. Differential endosomal sorting of a novel P2Y12 purinoreceptor mutant.

    Science.gov (United States)

    Cunningham, Margaret R; Nisar, Shaista P; Cooke, Alexandra E; Emery, Elizabeth D; Mundell, Stuart J

    2013-05-01

    P2Y12 receptor internalization and recycling play an essential role in ADP-induced platelet activation. Recently, we identified a patient with a mild bleeding disorder carrying a heterozygous mutation of P2Y12 (P341A) whose P2Y12 receptor recycling was significantly compromised. Using human cell line models, we identified key proteins regulating wild-type (WT) P2Y12 recycling and investigated P2Y12 -P341A receptor traffic. Treatment with ADP resulted in delayed Rab5-dependent internalization of P341A when compared with WT P2Y12 . While WT P2Y12 rapidly recycled back to the membrane via Rab4 and Rab11 recycling pathways, limited P341A recycling was observed, which relied upon Rab11 activity. Although minimal receptor degradation was evident, P341A was localized in Rab7-positive endosomes with considerable agonist-dependent accumulation in the trans-Golgi network (TGN). Rab7 activity is known to facilitate recruitment of retromer complex proteins to endosomes to transport cargo to the TGN. Here, we identified that P341A colocalized with Vps26; depletion of which blocked limited recycling and promoted receptor degradation. This study has identified key points of divergence in the endocytic traffic of P341A versus WT-P2Y12 . Given that these pathways are retained in human platelets, this research helps define the molecular mechanisms regulating P2Y12 receptor traffic and explain the compromised receptor function in the platelets of the P2Y12 -P341A-expressing patient. © 2013 John Wiley & Sons A/S.

  3. Shoc2 is targeted to late endosomes and required for Erk1/2 activation in EGF-stimulated cells.

    Directory of Open Access Journals (Sweden)

    Emilia Galperin

    Full Text Available Shoc2 is the putative scaffold protein that interacts with RAS and RAF, and positively regulates signaling to extracellular signal-regulated protein kinases 1 and 2 (ERK1/2. To elucidate the mechanism by which Shoc2 regulates ERK1/2 activation by the epidermal growth factor (EGF receptor (EGFR, we studied subcellular localization of Shoc2. Upon EGFR activation, endogenous Shoc2 and red fluorescent protein tagged Shoc2 were translocated from the cytosol to a subset of late endosomes containing Rab7. The endosomal recruitment of Shoc2 was blocked by overexpression of a GDP-bound H-RAS (N17S mutant and RNAi knockdown of clathrin, suggesting the requirement of RAS activity and clathrin-dependent endocytosis. RNAi depletion of Shoc2 strongly inhibited activation of ERK1/2 by low, physiological EGF concentrations, which was rescued by expression of wild-type recombinant Shoc2. In contrast, the Shoc2 (S2G mutant, that is myristoylated and found in patients with the Noonan-like syndrome, did not rescue ERK1/2 activation in Shoc2-depleted cells. Shoc2 (S2G was not located in late endosomes but was present on the plasma membrane and early endosomes. These data suggest that targeting of Shoc2 to late endosomes may facilitate EGFR-induced ERK activation under physiological conditions of cell stimulation by EGF, and therefore, may be involved in the spatiotemporal regulation of signaling through the RAS-RAF module.

  4. Laser ablation of dental calculus at 400 nm using a Ti:sapphire laser

    Science.gov (United States)

    Schoenly, Joshua E.; Seka, Wolf; Rechmann, Peter

    2009-02-01

    A Nd:YAG laser-pumped, frequency-doubled Ti:sapphire laser is used for selective ablation of calculus. The laser provides calculus removal. This is in stark contrast with tightly focused Gaussian beams that are energetically inefficient and lead to irreproducible results. Calculus is well ablated at high fluences >=2J/cm2 stalling occurs below this fluence because of photobleaching. Healthy hard tissue is not removed at fluences <=3 J/cm2.

  5. Microwave Ablation Compared with Radiofrequency Ablation for Breast Tissue in an Ex Vivo Bovine Udder Model

    International Nuclear Information System (INIS)

    Tanaka, Toshihiro; Westphal, Saskia; Isfort, Peter; Braunschweig, Till; Penzkofer, Tobias; Bruners, Philipp; Kichikawa, Kimihiko; Schmitz-Rode, Thomas; Mahnken, Andreas H.

    2012-01-01

    Purpose: To compare the effectiveness of microwave (MW) ablation with radiofrequency (RF) ablation for treating breast tissue in a nonperfused ex vivo model of healthy bovine udder tissue. Materials and Methods: MW ablations were performed at power outputs of 25W, 35W, and 45W using a 915-MHz frequency generator and a 2-cm active tip antenna. RF ablations were performed with a bipolar RF system with 2- and 3-cm active tip electrodes. Tissue temperatures were continuously monitored during ablation. Results: The mean short-axis diameters of the coagulation zones were 1.34 ± 0.14, 1.45 ± 0.13, and 1.74 ± 0.11 cm for MW ablation at outputs of 25W, 35W, and 45W. For RF ablation, the corresponding values were 1.16 ± 0.09 and 1.26 ± 0.14 cm with electrodes having 2- and 3-cm active tips, respectively. The mean coagulation volumes were 2.27 ± 0.65, 2.85 ± 0.72, and 4.45 ± 0.47 cm 3 for MW ablation at outputs of 25W, 35W, and 45W and 1.18 ± 0.30 and 2.29 ± 0.55 cm 3 got RF ablation with 2- and 3-cm electrodes, respectively. MW ablations at 35W and 45W achieved significantly longer short-axis diameters than RF ablations (P < 0.05). The highest tissue temperature was achieved with MW ablation at 45W (P < 0.05). On histological examination, the extent of the ablation zone in MW ablations was less affected by tissue heterogeneity than that in RF ablations. Conclusion: MW ablation appears to be advantageous with respect to the volume of ablation and the shape of the margin of necrosis compared with RF ablation in an ex vivo bovine udder.

  6. Percutaneous thermal ablation of renal neoplasms

    International Nuclear Information System (INIS)

    Tacke, J.; Mahnken, A.H.; Guenther, R.W.

    2005-01-01

    Due to modern examination techniques such as multidetector computed tomography and high-field magnetic resonance imaging, the detection rate of renal neoplasms is continually increasing. Even though tumors exceeding 4 cm in diameter rarely metastasize, all renal lesions that are possible neoplasms should be treated. Traditional treatment techniques include radical nephrectomy or nephron-sparing resection, which are increasingly performed laparoscopically. Modern thermal ablation techniques such as hyperthermal techniques like radiofrequency ablation RFA, laser induced thermal ablation LITT, focused ultrasound FUS and microwave therapy MW, as well as hypothermal techniques (cryotherapy) may be a useful treatment option for patients who are unfit for or refuse surgical resection. Cryotherapy is the oldest and best known thermal ablation technique and can be performed laparoscopically or percutaneously. Since subzero temperatures have no antistyptic effect, additional maneuvers must be performed to control bleeding. Percutaneous cryotherapy of renal tumors is a new and interesting method, but experience with it is still limited. Radiofrequency ablation is the most frequently used method. Modern probe design allows volumes between 2 and 5 cm in diameter to be ablated. Due to hyperthermal tract ablation, the procedure is deemed to be safe and has a low complication rate. Although there are no randomized comparative studies to open resection, the preliminary results for renal RFA are promising and show RFA to be superior to other thermal ablation techniques. Clinical success rates are over 90% for both, cryo- and radiofrequency ablation. Whereas laser induced thermal therapy is established in hepatic ablation, experience is minimal with respect to renal application. For lesions of more than 2 cm in diameter, additional cooling catheters are required. MR thermometry offers temperature control during ablation. Microwave ablation is characterized by small ablation volumes

  7. Endometrial ablation by rollerball electrocoagulation compared to uterine balloon thermal ablation. Technical and safety aspects.

    NARCIS (Netherlands)

    Zon-Rabelink, I.A.A. van; Vleugels, M.P.; Merkus, J.M.W.M.; Graaf, R.M. de

    2003-01-01

    OBJECTIVE: To compare two methods of endometrial ablation, hysteroscopic rollerball electrocoagulation (RBE) and non-hysteroscopic uterine balloon thermal (UBT) ablation (Thermachoice), regarding intra- and post-operative technical complications and safety aspects. STUDY DESIGN: A randomised

  8. Attitudes Towards Catheter Ablation for Atrial Fibrillation

    DEFF Research Database (Denmark)

    Vadmann, Henrik; Pedersen, Susanne S; Nielsen, Jens Cosedis

    2015-01-01

    BACKGROUND: Catheter ablation for atrial fibrillation (AF) is an important but expensive procedure that is the subject of some debate. Physicians´ attitudes towards catheter ablation may influence promotion and patient acceptance. This is the first study to examine the attitudes of Danish...... cardiologists towards catheter ablation for AF, using a nationwide survey. METHODS AND RESULTS: We developed a purpose-designed questionnaire to evaluate attitudes towards catheter ablation for AF that was sent to all Danish cardiologists (n = 401; response n = 272 (67.8%)). There was no association between...... attitudes towards ablation and the experience or age of the cardiologist with respect to patients with recurrent AF episodes with a duration of 7 days and/or need for cardioversion. The majority (69%) expected a recurrence of AF after catheter ablation in more than 30% of the cases...

  9. Lung Tumor Radiofrequency Ablation: Where Do We Stand?

    International Nuclear Information System (INIS)

    Baère, Thierry de

    2011-01-01

    Today, radiofrequency ablation (RFA) of primary and metastatic lung tumor is increasingly used. Because RFA is most often used with curative intent, preablation workup must be a preoperative workup. General anesthesia provides higher feasibility than conscious sedation. The electrode positioning must be performed under computed tomography for sake of accuracy. The delivery of RFA must be adapted to tumor location, with different impedances used when treating tumors with or without pleural contact. The estimated rate of incomplete local treatment at 18 months was 7% (95% confidence interval, 3–14) per tumor, with incomplete treatment depicted at 4 months (n = 1), 6 months (n = 2), 9 months (n = 2), and 12 months (n = 2). Overall survival and lung disease-free survival at 18 months were, respectively, 71 and 34%. Size is a key point for tumor selection because large size is predictive of incomplete local treatment and poor survival. The ratio of ablation volume relative to tumor volume is predictive of complete ablation. Follow-up computed tomography that relies on the size of the ablation zone demonstrates the presence of incomplete ablation. Positron emission tomography might be an interesting option. Chest tube placement for pneumothorax is reported in 8 to 12%. Alveolar hemorrhage and postprocedure hemoptysis occurred in approximately 10% of procedures and rarely required specific treatment. Death was mostly related to single-lung patients and hilar tumors. No modification of forced expiratory volume in the first second between pre- and post-RFA at 2 months was found. RFA in the lung provides a high local efficacy rate. The use of RFA as a palliative tool in combination with chemotherapy remains to be explored.

  10. Endometrial ablation in the management of abnormal uterine bleeding.

    Science.gov (United States)

    Laberge, Philippe; Leyland, Nicholas; Murji, Ally; Fortin, Claude; Martyn, Paul; Vilos, George; Leyland, Nicholas; Wolfman, Wendy; Allaire, Catherine; Awadalla, Alaa; Dunn, Sheila; Heywood, Mark; Lemyre, Madeleine; Marcoux, Violaine; Potestio, Frank; Rittenberg, David; Singh, Sukhbir; Yeung, Grace

    2015-04-01

    Abnormal uterine bleeding (AUB) is the direct cause of a significant health care burden for women, their families, and society as a whole. Up to 30% of women will seek medical assistance for the problem during their reproductive years. To provide current evidence-based guidelines on the techniques and technologies used in endometrial ablation (EA), a minimally invasive technique for the management of AUB of benign origin. Members of the guideline committee were selected on the basis of individual expertise to represent a range of practical and academic experience in terms of both location in Canada and type of practice, as well as subspecialty expertise and general background in gynaecology. The committee reviewed all available evidence in the English medical literature, including published guidelines, and evaluated surgical and patient outcomes for the various EA techniques. Recommendations were established by consensus. Published literature was retrieved through searches of MEDLINE and The Cochrane Library in 2013 and 2014 using appropriate controlled vocabulary and key words (endometrial ablation, hysteroscopy, menorrhagia, heavy menstrual bleeding, AUB, hysterectomy). RESULTS were restricted to systematic reviews, randomized control trials/controlled clinical trials, and observational studies written in English from January 2000 to November 2014. Searches were updated on a regular basis and incorporated in the guideline to December 2014. Grey (unpublished) literature was identifies through searching the websites of health technology assessment and health technology-related agencies, clinical practice guideline collections, clinical trial registries, and national and international medical specialty societies. The quality of evidence in this document was rated using the criteria described in the Report of the Canadian Task Force on Preventive Health Care (Table 1). This document reviews the evidence regarding the available techniques and technologies for EA

  11. Real-time monitoring of indium tin oxide laser ablation in liquid crystal display patterning

    International Nuclear Information System (INIS)

    Hong, M.H.; Lu, Y.F.; Meng, M.; Low, T.S.

    1998-01-01

    Audible acoustic wave detection is applied to investigate KrF excimer laser ablation of Indium Tin Oxide (ITO) thin film layer for Liquid Crystal Display (LCD) patterning. It is found that there is no acoustic wave generation if laser fluence is lower than ITO ablation threshold. For laser fluence higher than the threshold, audible acoustic wave will be detected due to shock wave generation during ITO laser ablation. The amplitude of the acoustic wave is closely related to the laser ablation rate. With more laser pulse applied, the amplitude is dropped to zero because the ITO layer is completely removed. However, if laser fluence is increased higher than ablation threshold for glass substrate, the amplitude is also dropped with pulse number but not to zero. It is due to laser ablation of ITO layer and glass substrate at the same time. Since the thickness of ITO layer is in a scale of 100 nm, laser interaction with glass substrate will happen even at the first pulse of higher laser fluence irradiation. Laser ablation induced ITO plasma emission spectrum in visible light region is analyzed by an Optical Multi-channel Analyzer (OMA). Specific spectral lines are In I (325.8, 410.2 and 451.1 nm) and In II 591.1 nm. Spectral intensities of 410.2 and 451.1 nm lines are selected to characterize the evolution of ITO plasma intensity with laser fluence and pulse number. It is found that the spectral intensities are reduced to zero with laser pulse number. It is also found that spectral lines other than ITO plasma will appear for laser fluence higher than ablation threshold for glass substrate. Threshold fluences for glass and ITO ablation are estimated for setting up a parameter window to control LCD patterning in real-time

  12. Radiofrequency Ablation of Hepatic Cysts : Case Report

    International Nuclear Information System (INIS)

    Lee, Ye Ri; Kim, Pyo Nyun

    2005-01-01

    Radiofrequency ablation has been frequently performed on intra-hepatic solid tumor, namely, hepatocellular carcinoma, metastatic tumor and cholangio carcinoma, for take the cure. But, the reports of radiofrequency ablation for intrahepatic simple cysts are few. In vitro experiment of animal and in vivo treatment for intrahepatic cysts of human had been reported in rare cases. We report 4 cases of radiofrequency ablation for symptomatic intrahepatic cysts

  13. Synthesis of Fe–Ni bimetallic nanoparticles from pixel target ablation: plume dynamics and surface characterization

    International Nuclear Information System (INIS)

    Niu Xiaoxu; Murray, Paul T.; Sarangan, Andrew

    2012-01-01

    A novel Fe–Ni bimetallic nanoparticle synthesis technique, denoted pixel target ablation, is reported. The technique entails ablating a thin film target consisting of patterned Fe and Ni pixels with a selected ratio using a KrF excimer laser. The laser energy breaks a known amount of target materials into metal atoms, which then form nanoparticles by recombination in the gas phase. Due to the nature of thin-film ablation, splashing of large particles was eliminated with the added benefit of minimizing nanoparticle agglomeration. Plume dynamics and surface characterizations were carried out to exploit the formation of Fe–Ni nanoparticles more fully. The composition was readily controlled by varying the initial relative amount of Fe and Ni target pixels. Synthesis of multi-element nanoparticles by pixel target ablation should be possible with any element combination that can be prepared as a thin-film target.

  14. Global Endometrial Ablation in the Presence of Essure® Microinserts

    Science.gov (United States)

    Aldape, Diana; Chudnoff, Scott G; Levie, Mark D

    2013-01-01

    Abnormal uterine bleeding (AUB) affects 30% of women at some time during their reproductive years and is one of the most common reasons a woman sees a gynecologist. Many women are turning to endometrial ablation to manage their AUB. This article reviews the data relating to the available endometrial ablation techniques performed with hysteroscopic sterilization, and focuses on data from patients who had Essure® (Conceptus, San Carlos, CA) coils placed prior to performance of endometrial ablation. Reviewed specifically are data regarding safety and efficacy of these two procedures when combined. Data submitted to the US Food and Drug Administration for the three devices currently approved are reviewed, as well as all published case series. Articles included were selected based on a PubMed search for endometrial ablation (also using the brand names of the different techniques currently available), hysteroscopic sterilization, and Essure. PMID:24358407

  15. Effect of Hashimoto thyroiditis on low-dose radioactive-iodine remnant ablation.

    Science.gov (United States)

    Kwon, Hyungju; Choi, June Young; Moon, Jae Hoon; Park, Hyo Jin; Lee, Won Woo; Lee, Kyu Eun

    2016-04-01

    Radioactive-iodine remnant ablation is an integral part of the papillary thyroid carcinoma (PTC) treatment. Although a minimum dose is usually recommended, there is controversy as to whether the low-dose (1100 MBq) radioactive-iodine remnant ablation is adequate for selected patients. A retrospective cohort study was conducted on 691 patients. Patients with no remnant thyroid on the follow-up whole body scan and low stimulated thyroglobulin (sTg) level (Hashimoto thyroiditis based on histopathology diagnosis (odds ratio [OR] = 3.23; p Hashimoto thyroiditis and elevated sTg are negative predictive factors for successful low-dose radioactive-iodine remnant ablation treatment. An appropriate risk-adjusted approach may improve the efficacy of radioactive-iodine remnant ablation treatment. © 2015 Wiley Periodicals, Inc. Head Neck 38: E730-E735, 2016. © 2015 Wiley Periodicals, Inc.

  16. Cryoballoon Catheter Ablation in Atrial Fibrillation

    Directory of Open Access Journals (Sweden)

    Cevher Ozcan

    2011-01-01

    Full Text Available Pulmonary vein isolation with catheter ablation is an effective treatment in patients with symptomatic atrial fibrillation refractory or intolerant to antiarrhythmic medications. The cryoballoon catheter was recently approved for this procedure. In this paper, the basics of cryothermal energy ablation are reviewed including its ability of creating homogenous lesion formation, minimal destruction to surrounding vasculature, preserved tissue integrity, and lower risk of thrombus formation. Also summarized here are the publications describing the clinical experience with the cryoballoon catheter ablation in both paroxysmal and persistent atrial fibrillation, its safety and efficacy, and discussions on the technical aspect of the cryoballoon ablation procedure.

  17. Differences in patient selection and outcomes between SilverHawk atherectomy and laser ablation in the treatment of femoropopliteal in-stent restenosis: a retrospective analysis from a single center.

    Science.gov (United States)

    Shammas, Nicolas W; Shammas, Gail A; Jerin, Michael

    2013-12-01

    To analyze differences in the application and outcomes of SilverHawk atherectomy (SH) and excimer laser ablation (ELA) in the treatment of femoropopliteal in-stent restenosis (ISR) in an unselected cohort of patients treated at a single center. Between January 2005 and June 2010, 81 consecutive patients (46 men; mean age 69.1 years, range 43-86) underwent directional atherectomy (41 SH, 40 ELA) for femoropopliteal ISR lesions. Data were reviewed retrospectively on procedural outcomes, major adverse events, and 1-year target lesion revascularization (TLR) obtained from medical records and supplemented with telephone calls. The primary endpoint was symptom-driven TLR at 1 year; secondary endpoints were death and amputation. ELA was utilized more frequently than SH in longer lesions (210.4±104 vs. 126.2±79.3 mm, respectively; p=0.001), subacute presentation (55% vs. 14.6%, p=0.001), TASC D lesions (47.5% vs. 12.2%), and in patients with more angiographic thrombus (42.5% vs. 4.9%, p=0.001). Final angiographic success (<30% residual narrowing post final treatment) was similar between ELA and SH (92.5% vs. 100%, respectively, p=0.12). Bailout stenting was significantly higher with ELA vs. SH (50.0% vs. 24.4%, p=0.022). At 1 year, TLR had occurred in 48.7% of the ELA patients vs. 31.7% of the SH cases (p=0.171). Regression analysis confirmed that SH was a predictor of TLR at 1 year (hazard ratio 2.679, 95% CI 1.015 to 7.073, p=0.047). Both SH and ELA continue to have a high TLR rate in treating ISR of the femoral and popliteal arteries. A higher rate of delayed failure is seen with SH and an earlier, steeper loss of TLR-free survival is seen with ELA.

  18. Two barcodes encoded by the type-1 PDZ and by phospho-Ser312 regulate retromer/WASH-mediated sorting of the ß1-adrenergic receptor from endosomes to the plasma membrane.

    Science.gov (United States)

    Nooh, Mohammed M; Bahouth, Suleiman W

    2017-01-01

    Recycling of the majority of agonist-internalized GPCR is dependent on a type I-PDZ "barcode" in their C-tail. The recycling of wild-type (WT) ß 1 -AR is also dependent on its default "type-1 PDZ barcode", but trafficking of the ß 1 -AR is inhibited when PKA or its substrate serine at position 312 (Ser 312 ) are inactivated. We tested the hypothesis that phospho-Ser 312 provided a second barcode for ß 1 -AR sorting from endosomes to the plasma membrane by determining the role of retromer/WASH complexes in ß 1 -AR trafficking. Recycling of WT ß 1 -AR or WT ß 2 -AR was dependent on targeting the retromer to endosomal membranes via SNX3 and rab7a, and on complexing the retromer to the WASH pentamer via the C-tail of FAM21 (FAM21 C ). These maneuvers however, did not inhibit the recycling of a phospho-Ser 312 ß 1 -AR mimic ((S312D) ß 1 -AR). Knockdown of the trans-acting PDZ protein sorting nexin27 (SNX27) inhibited the recycling of WT ß 1 -AR and WT ß 2 -AR, but had no effect on (S312D) ß 1 -AR∆PDZ or on phosphorylation of WT ß 1 -AR by PKA at Ser 312 . However, depletion of FKBP15, a FAM21 C -binding endosomal protein, selectively inhibited WT ß 1 -AR but not ß 2 -AR recycling, suggesting divergence might exist in GPCR trafficking roadmaps. These results indicate that two barcodes are involved in sorting WT ß 1 -AR out of early endosomes. The first and antecedent "barcode" was the "type-1 PDZ", followed by a second reversible "phospho-Ser 312 " verification "barcode". This organization allows tight regulation of ß 1 -AR density to signaling intensity in conditions associated with aberrant ß 1 -AR signaling such as in hypertension and heart failure. Copyright © 2016 Elsevier Inc. All rights reserved.

  19. Comparison of the Effectiveness of Ablative and Non-Ablative Fractional Laser Treatments for Early Stage Thyroidectomy Scars

    Directory of Open Access Journals (Sweden)

    Jin-Uk Jang

    2016-11-01

    Full Text Available BackgroundOpen thyroidectomy is conventionally performed at the anterior side of neck, which is a body part with a comparatively great degree of open exposure; due to this, postoperative scarring may cause distress in patients. We aimed to compare the effects of ablative and nonablative fractional laser treatments on thyroidectomy scars. We examined medical records in a retrospective manner and analyzed scars based on their digital images by using the modified Manchester Scar Scale (mMSS.MethodsBetween February 2012 and May 2013, 55 patients with thyroidectomy scars were treated with ablative (34 patients or nonablative (21 patients fractional laser. Each patient underwent 4 laser treatment sessions in 3–4 week intervals, 1–2 months postoperatively. Scar improvement was assessed using patient images and the mMSS scale.ResultsThe mean decrease in scar score was 3.91 and 3.47 in the ablative and nonablative groups, respectively; the reduction between 2 groups did not exhibit any significant difference (P=0.16. We used the scale once again to individually evaluate scar attributes. The nonablative group accounted for a considerably higher color score value (P=0.03; the ablative group accounted for a considerably higher contour score value (P<0.01. Patient satisfaction was high and no complications occurred.ConclusionsBoth types of fractional laser treatments can be used successfully for thyroidectomy scar treatment with minimal complications; however, results indicate that higher effectiveness may be obtained from the use of ablative and nonablative lasers for hypertrophic scars and early erythematous scars, respectively. Therefore, the appropriate laser for scar treatment should be selected according to its specific characteristics.

  20. Solo/Trio8, a membrane-associated short isoform of Trio, modulates endosome dynamics and neurite elongation.

    Science.gov (United States)

    Sun, Ying-Jie; Nishikawa, Kaori; Yuda, Hideki; Wang, Yu-Lai; Osaka, Hitoshi; Fukazawa, Nobuna; Naito, Akira; Kudo, Yoshihisa; Wada, Keiji; Aoki, Shunsuke

    2006-09-01

    With DNA microarrays, we identified a gene, termed Solo, that is downregulated in the cerebellum of Purkinje cell degeneration mutant mice. Solo is a mouse homologue of rat Trio8-one of multiple Trio isoforms recently identified in rat brain. Solo/Trio8 contains N-terminal sec14-like and spectrin-like repeat domains followed by a single guanine nucleotide exchange factor 1 (GEF1) domain, but it lacks the C-terminal GEF2, immunoglobulin-like, and kinase domains that are typical of Trio. Solo/Trio8 is predominantly expressed in Purkinje neurons of the mouse brain, and expression begins following birth and increases during Purkinje neuron maturation. We identified a novel C-terminal membrane-anchoring domain in Solo/Trio8 that is required for enhanced green fluorescent protein-Solo/Trio8 localization to early endosomes (positive for both early-endosome antigen 1 [EEA1] and Rab5) in COS-7 cells and primary cultured neurons. Solo/Trio8 overexpression in COS-7 cells augmented the EEA1-positive early-endosome pool, and this effect was abolished via mutation and inactivation of the GEF domain or deletion of the C-terminal membrane-anchoring domain. Moreover, primary cultured neurons transfected with Solo/Trio8 showed increased neurite elongation that was dependent on these domains. These results suggest that Solo/Trio8 acts as an early-endosome-specific upstream activator of Rho family GTPases for neurite elongation of developing Purkinje neurons.

  1. Augmented internalisation of ferroportin to late endosomes impairs iron uptake by enterocyte-like IEC-6 cells.

    Science.gov (United States)

    Oates, Phillip S; Thomas, Carla

    2005-08-01

    Absorption of iron occurs by duodenal enterocytes, involving uptake by the divalent metal transporter-1 (DMT1) and release by ferroportin. Ferroportin responds to the hepatocyte-produced 25-amino-acid-peptide hepcidin-25 by undergoing internalisation to late endosomes that impair iron release. Ferroportin is also expressed on the apical membrane of polarised Caco-2 cells, rat intestinal cells and in IEC-6 cells (an intestinal epithelial cell line). A blocking antibody to ferroportin also impairs the uptake, but not the release, of iron. In this study IEC-6 cells were used to study the mechanism of impairment or recovery from impairment produced by the blocking antibody and the fate of DMT1 and ferroportin. Uptake of 1 muM Fe(II) was studied by adding the antibody from time 0 and after adding or removing the antibody once a steady state had been reached. Surface binding, maximum iron transport rate V(max) and transporter affinity (K(m)) were measured after impairment of iron uptake. Ferroportin and DMT1 distribution were assessed by immunofluorescence microscopy. Antibody-mediated impairment, or recovery from impairment, of Fe(II) uptake occurred within minutes. Impairment was lost when the antibody was combined with the immunizing peptide. DMT1 and ferroportin undergo internalisation to late endosomes and, in the presence of the antibody, augmented internalisation of DMT1 and ferroportin caused swelling of late endosomes. Surface binding of Fe(II) and iron transport V(max) were reduced by 50%, indicating that the antibody removed membrane-bound DMT1. The ferroportin antibody induced rapid turnover of membrane ferroportin and DMT1 and its internalisation to late endosomes, resulting in impaired Fe(II) uptake.

  2. Extracellular anti-angiogenic proteins augment an endosomal protein trafficking pathway to reach mitochondria and execute apoptosis in HUVECs.

    Science.gov (United States)

    Chen, Mo; Qiu, Tao; Wu, Jiajie; Yang, Yang; Wright, Graham D; Wu, Min; Ge, Ruowen

    2018-03-09

    Classic endocytosis destinations include the recycling endosome returning to the plasma membrane or the late endosome (LE) merging with lysosomes for cargo degradation. However, the anti-angiogenic proteins angiostatin and isthmin, are endocytosed and trafficked to mitochondria (Mito) to execute apoptosis of endothelial cells. How these extracellular proteins reach mitochondria remains a mystery. Through confocal and super-resolution fluorescent microscopy, we demonstrate that angiostatin and isthmin are trafficked to mitochondria through the interaction between LE and Mito. Using purified organelles, the LE-Mito interaction is confirmed through in vitro lipid-fusion assay, as well as single vesicle total internal reflection fluorescent microscopy. LE-Mito interaction enables the transfer of not only lipids but also proteins from LE to Mito. Angiostatin and isthmin augment this endosomal protein trafficking pathway and make use of it to reach mitochondria to execute apoptosis. Cell fractionation and biochemical analysis identified that the cytosolic scaffold protein Na+/H+ exchanger regulatory factor 1 (NHERF1) associated with LE and the t-SNARE protein synaptosome-associated protein 25 kDa (SNAP25) associated with Mito form an interaction complex to facilitate LE-Mito interaction. Proximity ligation assay coupled with fluorescent microscopy showed that both NHERF1 and SNAP25 are located at the contacting face between LE and Mito. RNAi knockdown of either NHERF1 or SNAP25 suppressed not only the mitochondrial trafficking of angiostatin and isthmin but also their anti-angiogenic and pro-apoptotic functions. Hence, this study reveals a previously unrealized endosomal protein trafficking pathway from LE to Mito that allows extracellular proteins to reach mitochondria and execute apoptosis.

  3. Amyloid precursor protein and endosomal-lysosomal dysfunction in Alzheimer's disease: inseparable partners in a multifactorial disease.

    Science.gov (United States)

    Nixon, Ralph A

    2017-07-01

    Abnormalities of the endosomal-lysosomal network (ELN) are a signature feature of Alzheimer's disease (AD). These include the earliest known cytopathology that is specific to AD and that affects endosomes and induces the progressive failure of lysosomes, each of which are directly linked by distinct mechanisms to neurodegeneration. The origins of ELN dysfunction and β-amyloidogenesis closely overlap, which reflects their common genetic basis, the established early involvement of endosomes and lysosomes in amyloid precursor protein (APP) processing and clearance, and the pathologic effect of certain APP metabolites on ELN functions. Genes that promote β-amyloidogenesis in AD (APP, PSEN1/2, and APOE4) have primary effects on ELN function. The importance of primary ELN dysfunction to pathogenesis is underscored by the mutations in more than 35 ELN-related genes that, thus far, are known to cause familial neurodegenerative diseases even though different pathogenic proteins may be involved. In this article, I discuss growing evidence that implicates AD gene-driven ELN disruptions as not only the antecedent pathobiology that underlies β-amyloidogenesis but also as the essential partner with APP and its metabolites that drive the development of AD, including tauopathy, synaptic dysfunction, and neurodegeneration. The striking amelioration of diverse deficits in animal AD models by remediating ELN dysfunction further supports a need to integrate APP and ELN relationships, including the role of amyloid-β, into a broader conceptual framework of how AD arises, progresses, and may be effectively therapeutically targeted.-Nixon, R. A. Amyloid precursor protein and endosomal-lysosomal dysfunction in Alzheimer's disease: inseparable partners in a multifactorial disease. © FASEB.

  4. Effects of Arsenic Trioxide on Radiofrequency Ablation of VX2 Liver Tumor: Intraarterial versus Intravenous Administration

    Energy Technology Data Exchange (ETDEWEB)

    Seong, Nak Jong; Yoon, Chang Jin; Kang, Sung Gwon [Seoul National University Bundang Hospital, Seongnam (Korea, Republic of); Chung, Jin Wook; Kim, Hyo Cheol; Park, Jae Hyung [Seoul National University Hospital, Seoul (Korea, Republic of)

    2012-03-15

    Arsenic trioxide (As{sub 2}O{sub 3}) can be used as a possible pharmaceutical alternative that augments radiofrequency (RF) ablation by reducing tumor blood flow. The aim of this study was to assess the effect of intraarterial and intravenous administration of As{sub 2}O{sub 3} on RF-induced ablation in an experimentally induced liver tumor. VX2 carcinoma was grown in the livers of 30 rabbits. As{sub 2}O{sub 3} (1 mg/kg) was administered through the hepatic artery (n = 10, group A) or ear vein (n = 10, group B), 30 minutes before RF ablation (125 mA {+-} 35; 90 {+-} 5 degrees Celsius). As a control group, 10 rabbits were treated with RF ablation alone (group C). RF was intentionally applied to the peripheral margin of the tumor so that ablation can cover the tumor and adjacent hepatic parenchyma. Ablation areas of the tumor and adjacent parenchymal changes among three groups were compared by the Kruskal-Wallis and Mann-Whitney U test. The overall ablation areas were 156 {+-} 28.9 mm{sup 2} (group A), 119 {+-} 31.7 (group B), and 92 {+-} 17.4 (group C, p < 0.04). The ablation area of the tumor was significantly larger in group A (73 {+-} 19.7 mm{sup 2}) than both group B (50 {+-} 19.4, p = 0.02) and group C (28 {+-} 2.2, p < 0.01). The ratios of the tumoral ablation area to the overall ablation area were larger in group A (47 {+-} 10.5%) than that of the other groups (42 {+-} 7.3% in group B and 32 {+-} 5.6% in group C) (p < 0.03). Radiofrequency-induced ablation area can be increased with intraarterial or intravenous administration of As{sub 2}O{sub 3}. The intraarterial administration of As{sub 2}O{sub 3} seems to be helpful for the selective ablation of the tumor.

  5. Increase in Volume of Ablation Zones during Follow-up Is Highly Suggestive of Ablation Site Recurrence in Colorectal Liver Metastases Treated with Radiofrequency Ablation

    NARCIS (Netherlands)

    Kele, Petra G.; de Jong, Koert P.; van der Jagt, Eric J.

    Purpose: To test the hypothesis that volume changes of ablation zones (AZs) on successive computed tomography (CT) scans could predict ablation site recurrences (ASRs) in patients with colorectal liver metastases treated by radiofrequency (RF) ablation. Materials and Methods: RF ablation was

  6. Laparoscopic Ultrasound-Guided Radiofrequency Ablation of Uterine Fibroids

    International Nuclear Information System (INIS)

    Milic, Andrea; Asch, Murray R.; Hawrylyshyn, Peter A.; Allen, Lisa M.; Colgan, Terence J.; Kachura, John R.; Hayeems, Eran B.

    2006-01-01

    Four patients with symptomatic uterine fibroids measuring less than 6 cm underwent laparoscopic ultrasound-guided radiofrequency ablation (RFA) using multiprobe-array electrodes. Follow-up of the treated fibroids was performed with gadolinium-enhanced magnetic resonance imaging (MRI) and patients' symptoms were assessed by telephone interviews. The procedure was initially technically successful in 3 of the 4 patients and MRI studies at 1 month demonstrated complete fibroid ablation. Symptom improvement, including a decrease in menstrual bleeding and pain, was achieved in 2 patients at 3 months. At 7 months, 1 of these 2 patients experienced symptom worsening which correlated with recurrent fibroid on MRI. The third, initially technically successfully treated patient did not experience any symptom relief after the procedure and was ultimately diagnosed with adenomyosis. Our preliminary results suggest that RFA is a technically feasible treatment for symptomatic uterine fibroids in appropriately selected patients

  7. Role of ventricular tachycardia ablation in arrhythmogenic right ventricular cardiomyopathy

    Directory of Open Access Journals (Sweden)

    Alberto Cipriani

    2017-11-01

    Full Text Available Arrhythmogenic right ventricular cardiomyopathy (ARVC is characterized by progressive fibro-fatty replacement of the myocardium that represents the substrate for recurrent sustained ventricular tachycardia (VT. These arrhythmias characterize the clinical course of a sizeable proportion of patients and have significant implications for their quality of life and long-term prognosis. Antiarrhythmic drugs are often poorly tolerated and usually provide incomplete control of arrhythmia relapses. Catheter ablation is a potentially effective strategy to treat frequent VT episodes and ICD shocks in ARVC patients. The aims of this review are to discuss the electrophysiological and electroanatomic substrates of ventricular tachycardia in patients with ARVC and to analyze the role of catheter ablation in their management with particular reference to selection of patients, technical issues, potential complications and outcomes.

  8. Effective photo-enhancement of cellular activity of fluorophore-octaarginine antisense PNA conjugates correlates with singlet oxygen formation, endosomal escape and chromophore lipophilicity

    DEFF Research Database (Denmark)

    Yarani, Reza; Shiraishi, Takehiko; Nielsen, Peter E.

    2018-01-01

    Photochemical internalization (PCI) is a cellular drug delivery method based on the generation of light-induced reactive oxygen species (ROS) causing damage to the endosomal membrane and thereby resulting in drug release to the cytoplasm. In our study a series of antisense fluorophore octaarginin...... indicate that efficient photodynamic endosomal escape is strongly dependent on the quantum yield for photochemical singlet oxygen formation, photostability as well as the lipophilicity of the chromophore....

  9. Niclosamide is a proton carrier and targets acidic endosomes with broad antiviral effects.

    Science.gov (United States)

    Jurgeit, Andreas; McDowell, Robert; Moese, Stefan; Meldrum, Eric; Schwendener, Reto; Greber, Urs F

    2012-01-01

    Viruses use a limited set of host pathways for infection. These pathways represent bona fide antiviral targets with low likelihood of viral resistance. We identified the salicylanilide niclosamide as a broad range antiviral agent targeting acidified endosomes. Niclosamide is approved for human use against helminthic infections, and has anti-neoplastic and antiviral effects. Its mode of action is unknown. Here, we show that niclosamide, which is a weak lipophilic acid inhibited infection with pH-dependent human rhinoviruses (HRV) and influenza virus. Structure-activity studies showed that antiviral efficacy and endolysosomal pH neutralization co-tracked, and acidification of the extracellular medium bypassed the virus entry block. Niclosamide did not affect the vacuolar H(+)-ATPase, but neutralized coated vesicles or synthetic liposomes, indicating a proton carrier mode-of-action independent of any protein target. This report demonstrates that physico-chemical interference with host pathways has broad range antiviral effects, and provides a proof of concept for the development of host-directed antivirals.

  10. Niclosamide is a proton carrier and targets acidic endosomes with broad antiviral effects.

    Directory of Open Access Journals (Sweden)

    Andreas Jurgeit

    Full Text Available Viruses use a limited set of host pathways for infection. These pathways represent bona fide antiviral targets with low likelihood of viral resistance. We identified the salicylanilide niclosamide as a broad range antiviral agent targeting acidified endosomes. Niclosamide is approved for human use against helminthic infections, and has anti-neoplastic and antiviral effects. Its mode of action is unknown. Here, we show that niclosamide, which is a weak lipophilic acid inhibited infection with pH-dependent human rhinoviruses (HRV and influenza virus. Structure-activity studies showed that antiviral efficacy and endolysosomal pH neutralization co-tracked, and acidification of the extracellular medium bypassed the virus entry block. Niclosamide did not affect the vacuolar H(+-ATPase, but neutralized coated vesicles or synthetic liposomes, indicating a proton carrier mode-of-action independent of any protein target. This report demonstrates that physico-chemical interference with host pathways has broad range antiviral effects, and provides a proof of concept for the development of host-directed antivirals.

  11. Smart DNA vectors based on cyclodextrin polymers: compaction and endosomal release.

    Science.gov (United States)

    Wintgens, Véronique; Leborgne, Christian; Baconnais, Sonia; Burckbuchler, Virginie; Le Cam, Eric; Scherman, Daniel; Kichler, Antoine; Amiel, Catherine

    2012-02-01

    Neutral β-cyclodextrin polymers (polyβCD) associated with cationic adamantyl derivatives (Ada) can be used to deliver plasmid DNA into cells. In absence of an endosomolytic agent, transfection efficiency remains low because most complexes are trapped in the endosomal compartment. We asked whether addition of an imidazole-modified Ada can increase efficiency of polyβCD/cationic Ada-based delivery system. We synthesized two adamantyl derivatives: Ada5, which has a spacer arm between the Ada moiety and a bi-cationic polar head group, and Ada6, which presents an imidazole group. Strength of association between polyβCD and Ada derivatives was evaluated by fluorimetric titration. Gel mobility shift assay, zeta potential, and dark field transmission electron microscopy experiments demonstrated the system allowed for efficient DNA compaction. In vitro transfection experiments performed on HepG2 and HEK293 cells revealed the quaternary system polyβCD/Ada5/Ada6/DNA has efficiency comparable to cationic lipid DOTAP. We successfully designed fine-tuned DNA vectors based on cyclodextrin polymers combined with two new adamantyl derivatives, leading to significant transfection associated with low toxicity.

  12. Apical transport of influenza A virus ribonucleoprotein requires Rab11-positive recycling endosome.

    Directory of Open Access Journals (Sweden)

    Fumitaka Momose

    Full Text Available Influenza A virus RNA genome exists as eight-segmented ribonucleoprotein complexes containing viral RNA polymerase and nucleoprotein (vRNPs. Packaging of vRNPs and virus budding take place at the apical plasma membrane (APM. However, little is known about the molecular mechanisms of apical transport of newly synthesized vRNP. Transfection of fluorescent-labeled antibody and subsequent live cell imaging revealed that punctate vRNP signals moved along microtubules rapidly but intermittently in both directions, suggestive of vesicle trafficking. Using a series of Rab family protein, we demonstrated that progeny vRNP localized to recycling endosome (RE in an active/GTP-bound Rab11-dependent manner. The vRNP interacted with Rab11 through viral RNA polymerase. The localization of vRNP to RE and subsequent accumulation to the APM were impaired by overexpression of Rab binding domains (RBD of Rab11 family interacting proteins (Rab11-FIPs. Similarly, no APM accumulation was observed by overexpression of class II Rab11-FIP mutants lacking RBD. These results suggest that the progeny vRNP makes use of Rab11-dependent RE machinery for APM trafficking.

  13. Endosomal sorting complexes required for ESCRTing cells toward death during neurogenesis, neurodevelopment and neurodegeneration.

    Science.gov (United States)

    Kaul, Zenia; Chakrabarti, Oishee

    2018-03-25

    The endosomal sorting complexes required for transport (ESCRT) proteins help in the recognition, sorting and degradation of ubiquitinated cargoes from the cell surface, long-lived proteins or aggregates, and aged organelles present in the cytosol. These proteins take part in the endo-lysosomal system of degradation. The ESCRT proteins also play an integral role in cytokinesis, viral budding and mRNA transport. Many neurodegenerative diseases are caused by toxic accumulation of cargo in the cell, which causes stress and ultimately leads to neuronal death. This accumulation of cargo occurs because of defects in the endo-lysosomal degradative pathway-loss of function of ESCRTs has been implicated in this mechanism. ESCRTs also take part in many survival processes, lack of which can culminate in neuronal cell death. While the role played by the ESCRT proteins in maintaining healthy neurons is known, their role in neurodegenerative diseases is still poorly understood. In this review, we highlight the importance of ESCRTs in maintaining healthy neurons and then suggest how perturbations in many of the survival mechanisms governed by these proteins could eventually lead to cell death; quite often these correlations are not so obviously laid out. Extensive neuronal death eventually culminates in neurodegeneration. © 2018 John Wiley & Sons A/S. Published by John Wiley & Sons Ltd.

  14. Aggregation of nanoparticles in endosomes and lysosomes produces surface-enhanced Raman spectroscopy

    Science.gov (United States)

    Lucas, Leanne J.; Chen, Xiaoke K.; Smith, Aaron J.; Korbelik, Mladen; Zeng, Haishan; Lee, Patrick W. K.; Hewitt, Kevin Cecil

    2015-01-01

    The purpose of this study was to explore the use of surface-enhanced Raman spectroscopy (SERS) to image the distribution of epidermal growth factor receptor (EGFR) in cells. To accomplish this task, 30-nm gold nanoparticles (AuNPs) tagged with antibodies to EGFR (1012 per mL) were incubated with cells (106 per mL) of the A431 human epidermoid carcinoma and normal human bronchial epithelial cell lines. Using the 632.8-nm excitation line of a He-Ne laser, Raman spectroscopy measurements were performed using a point mapping scheme. Normal cells show little to no enhancement. SERS signals were observed inside the cytoplasm of A431 cells with an overall enhancement of 4 to 7 orders of magnitude. Raman intensity maps of the 1450 and 1583 cm-1 peaks correlate well with the expected distribution of EGFR and AuNPs, aggregated following uptake by endosomes and lysosomes. Spectral features from tyrosine and tryptophan residues dominate the SERS signals.

  15. Peroxisomes, lipid droplets, and endoplasmic reticulum "hitchhike" on motile early endosomes.

    Science.gov (United States)

    Guimaraes, Sofia C; Schuster, Martin; Bielska, Ewa; Dagdas, Gulay; Kilaru, Sreedhar; Meadows, Ben R A; Schrader, Michael; Steinberg, Gero

    2015-12-07

    Intracellular transport is mediated by molecular motors that bind cargo to be transported along the cytoskeleton. Here, we report, for the first time, that peroxisomes (POs), lipid droplets (LDs), and the endoplasmic reticulum (ER) rely on early endosomes (EEs) for intracellular movement in a fungal model system. We show that POs undergo kinesin-3- and dynein-dependent transport along microtubules. Surprisingly, kinesin-3 does not colocalize with POs. Instead, the motor moves EEs that drag the POs through the cell. PO motility is abolished when EE motility is blocked in various mutants. Most LD and ER motility also depends on EE motility, whereas mitochondria move independently of EEs. Covisualization studies show that EE-mediated ER motility is not required for PO or LD movement, suggesting that the organelles interact with EEs independently. In the absence of EE motility, POs and LDs cluster at the growing tip, whereas ER is partially retracted to subapical regions. Collectively, our results show that moving EEs interact transiently with other organelles, thereby mediating their directed transport and distribution in the cell. © 2015 Guimaraes et al.

  16. Vascular endothelial growth factor A-stimulated signaling from endosomes in primary endothelial cells.

    Science.gov (United States)

    Fearnley, Gareth W; Smith, Gina A; Odell, Adam F; Latham, Antony M; Wheatcroft, Stephen B; Harrison, Michael A; Tomlinson, Darren C; Ponnambalam, Sreenivasan

    2014-01-01

    The vascular endothelial growth factor A (VEGF-A) is a multifunctional cytokine that stimulates blood vessel sprouting, vascular repair, and regeneration. VEGF-A binds to VEGF receptor tyrosine kinases (VEGFRs) and stimulates intracellular signaling leading to changes in vascular physiology. An important aspect of this phenomenon is the spatiotemporal coordination of VEGFR trafficking and intracellular signaling to ensure that VEGFR residence in different organelles is linked to downstream cellular outputs. Here, we describe a series of assays to evaluate the effects of VEGF-A-stimulated intracellular signaling from intracellular compartments such as the endosome-lysosome system. These assays include the initial isolation and characterization of primary human endothelial cells, performing reverse genetics for analyzing protein function; methods used to study receptor trafficking, signaling, and proteolysis; and assays used to measure changes in cell migration, proliferation, and tubulogenesis. Each of these assays has been exemplified with studies performed in our laboratories. In conclusion, we describe necessary techniques for studying the role of VEGF-A in endothelial cell function. © 2014 Elsevier Inc. All rights reserved.

  17. Therapeutic efficacy of percutaneous radiofrequency ablation versus microwave ablation for hepatocellular carcinoma.

    Directory of Open Access Journals (Sweden)

    Lei Zhang

    Full Text Available The aim of this study was to investigate the therapeutic efficacy of percutaneous radiofrequency (RF ablation versus microwave (MW ablation for hepatocellular carcinoma (HCC measuring ≤ 5 cm in greatest diameter. From January 2006 to December 2006, 78 patients had undergone RF ablation whereas 77 had undergone MW ablation. Complete ablation (CA, local tumour progression (LTP and distant recurrence (DR were compared. The overall survival curves were calculated with the Kaplan-Meier technique and compared with the log-rank test. The CA rate was 83.4% (78/93 for RF ablation and 86.7%(91/105 for MW ablation. The LTP rate was 11.8% (11/93 for RF ablation and 10.5% (11/105 for MW ablation. DR was found in 51 (65.4% in the RF ablation and 62 (80.5% in the MW ablation. There was no significant difference in the 1-, 3-, and 5-year overall survival rates (P = 0.780 and the 1-, 3-, and 5-year disease-free survival rates (P = 0.123 between RF and MW ablation. At subgroup analyses, for patients with tumors ≤ 3.0 cm, there was no significant difference in the 1-, 3-, and 5-year overall survival rates (P = 0.067 and the corresponding disease-free survival rates(P = 0.849. For patients with tumor diameters of 3.1-5.0 cm, the 1-, 3-, and 5-year overall survival rates were 87.1%, 61.3%, and 40.1% for RF ablation and 85.4%, 36.6%, and 22% for MW ablation, with no significant difference (P = 0.068. The corresponding disease-free survival rates were 74.2%, 54.8%, and 45.2% for the RF ablation group and 53.3%, 26.8%, and 17.1% for the MW ablation group. The disease-free survival curve for the RF ablation group was significantly better than that for the MW ablation group (P = 0.018. RF ablation and MW ablation are both effective methods in treating hepatocellular carcinomas, with no significant differences in CA, LTP, DR, and overall survival.

  18. Acid-sensing ion channel (ASIC) 4 predominantly localizes to an early endosome-related organelle upon heterologous expression.

    Science.gov (United States)

    Schwartz, Verena; Friedrich, Katharina; Polleichtner, Georg; Gründer, Stefan

    2015-12-15

    Acid-sensing ion channels (ASICs) are voltage-independent proton-gated amiloride sensitive sodium channels, belonging to the DEG/ENaC gene family. Six different ASICs have been identified (ASIC1a, ASIC1b, ASIC2a, ASIC2b, ASIC3, ASIC4) that are activated by a drop in extracellular pH, either as homo- or heteromers. An exception is ASIC4, which is not activated by protons as a homomer and which does not contribute to functional heteromeric ASICs. Insensitivity of ASIC4 to protons and its comparatively low sequence identity to other ASICs (45%) raises the question whether ASIC4 may have different functions than other ASICs. In this study, we therefore investigated the subcellular localization of ASIC4 in heterologous cell lines, which revealed a surprising accumulation of the channel in early endosome-related vacuoles. Moreover, we identified an unique amino-terminal motif as important for forward-trafficking from the ER/Golgi to the early endosome-related compartment. Collectively, our results show that heterologously expressed ASIC4 predominantly resides in an intracellular endosomal compartment.

  19. African Swine Fever Virus Undergoes Outer Envelope Disruption, Capsid Disassembly and Inner Envelope Fusion before Core Release from Multivesicular Endosomes.

    Directory of Open Access Journals (Sweden)

    Bruno Hernáez

    2016-04-01

    Full Text Available African swine fever virus (ASFV is a nucleocytoplasmic large DNA virus (NCLDV that causes a highly lethal disease in domestic pigs. As other NCLDVs, the extracellular form of ASFV possesses a multilayered structure consisting of a genome-containing nucleoid successively wrapped by a thick protein core shell, an inner lipid membrane, an icosahedral protein capsid and an outer lipid envelope. This structural complexity suggests an intricate mechanism of internalization in order to deliver the virus genome into the cytoplasm. By using flow cytometry in combination with pharmacological entry inhibitors, as well as fluorescence and electron microscopy approaches, we have dissected the entry and uncoating pathway used by ASFV to infect the macrophage, its natural host cell. We found that purified extracellular ASFV is internalized by both constitutive macropinocytosis and clathrin-mediated endocytosis. Once inside the cell, ASFV particles move from early endosomes or macropinosomes to late, multivesicular endosomes where they become uncoated. Virus uncoating requires acidic pH and involves the disruption of the outer membrane as well as of the protein capsid. As a consequence, the inner viral membrane becomes exposed and fuses with the limiting endosomal membrane to release the viral core into the cytosol. Interestingly, virus fusion is dependent on virus protein pE248R, a transmembrane polypeptide of the inner envelope that shares sequence similarity with some members of the poxviral entry/fusion complex. Collective evidence supports an entry model for ASFV that might also explain the uncoating of other multienveloped icosahedral NCLDVs.

  20. Caveolin targeting to late endosome/lysosomal membranes is induced by perturbations of lysosomal pH and cholesterol content

    Science.gov (United States)

    Mundy, Dorothy I.; Li, Wei Ping; Luby-Phelps, Katherine; Anderson, Richard G. W.

    2012-01-01

    Caveolin-1 is an integral membrane protein of plasma membrane caveolae. Here we report that caveolin-1 collects at the cytosolic surface of lysosomal membranes when cells are serum starved. This is due to an elevation of the intralysosomal pH, since ionophores and proton pump inhibitors that dissipate the lysosomal pH gradient also trapped caveolin-1 on late endosome/lysosomes. Accumulation is both saturable and reversible. At least a portion of the caveolin-1 goes to the plasma membrane upon reversal. Several studies suggest that caveolin-1 is involved in cholesterol transport within the cell. Strikingly, we find that blocking cholesterol export from lysosomes with progesterone or U18666A or treating cells with low concentrations of cyclodextrin also caused caveolin-1 to accumulate on late endosome/lysosomal membranes. Under these conditions, however, live-cell imaging shows cavicles actively docking with lysosomes, suggesting that these structures might be involved in delivering caveolin-1. Targeting of caveolin-1 to late endosome/lysosomes is not observed normally, and the degradation rate of caveolin-1 is not altered by any of these conditions, indicating that caveolin-1 accumulation is not a consequence of blocked degradation. We conclude that caveolin-1 normally traffics to and from the cytoplasmic surface of lysosomes during intracellular cholesterol trafficking. PMID:22238363

  1. Gallbladder ablation by radiologic intervention

    International Nuclear Information System (INIS)

    Becker, C.D.; Quenville, N.F.; Burhenne, H.J.

    1988-01-01

    Thirty pigs underwent cystic duct occlusion by means of transcatheter endoluminal bipolar radiofrequency electrocoagulation under fluoroscopic guidance. Twenty animals subsequently underwent gallbladder ablation with alcohol and sotradecol; ten animals served as controls. Serial histologic blocking of the common bile duct, cystic duct, and gallbladder in all animals revealed no adverse effects of the sclerosants on the bile ducts or the structures adjacent to the gallbladder. The combination of 95% alcohol plus 3% sotradecol resulted in necrosis of the gallbladder mucosa within 2 weeks (two of two animals) and complete eradication of the mucosa and fibrotic obliteration of the gallbladder lumen within 8 weeks (six of eight animals)

  2. Hyperkalaemia after radiofrequency ablation of hepatocellular carcinoma

    NARCIS (Netherlands)

    Verhoevena, BH; Haagsma, EB; Appeltans, BMG; Slooff, MJH; de Jong, KP

    Radiofrequency ablation of liver tumours is a useful therapy for otherwise unresectable tumours. The complication rate is said to be low. In this case report we describe hyperkalaemia after radiofrequency ablation of a hepatocellular carcinoma in a patient with end-stage renal insufficiency. (C)

  3. Testing and evaluation of light ablation decontamination

    International Nuclear Information System (INIS)

    Demmer, R.L.; Ferguson, R.L.

    1994-10-01

    This report details the testing and evaluation of light ablation decontamination. It details WINCO contracted research and application of light ablation efforts by Ames Laboratory. Tests were conducted with SIMCON (simulated contamination) coupons and REALCON (actual radioactive metal coupons) under controlled conditions to compare cleaning effectiveness, speed and application to plant process type equipment

  4. Ablative Laser Propulsion: An Update, Part I

    International Nuclear Information System (INIS)

    Pakhomov, Andrew V.; Cohen, Timothy; Lin Jun; Thompson, M. Shane; Herren, Kenneth A.

    2004-01-01

    This paper presents an updated review of studies on Ablative Laser Propulsion conducted by the Laser Propulsion Group (LPG) at the University of Alabama in Huntsville. In particular, we describe the newest results of our experimental study of specific impulses and coupling coefficients achieved by double-pulsed ablation of graphite, aluminum, copper and lead targets

  5. Modeling the signaling endosome hypothesis: Why a drive to the nucleus is better than a (random walk

    Directory of Open Access Journals (Sweden)

    Howe Charles L

    2005-10-01

    Full Text Available Abstract Background Information transfer from the plasma membrane to the nucleus is a universal cell biological property. Such information is generally encoded in the form of post-translationally modified protein messengers. Textbook signaling models typically depend upon the diffusion of molecular signals from the site of initiation at the plasma membrane to the site of effector function within the nucleus. However, such models fail to consider several critical constraints placed upon diffusion by the cellular milieu, including the likelihood of signal termination by dephosphorylation. In contrast, signaling associated with retrogradely transported membrane-bounded organelles such as endosomes provides a dephosphorylation-resistant mechanism for the vectorial transmission of molecular signals. We explore the relative efficiencies of signal diffusion versus retrograde transport of signaling endosomes. Results Using large-scale Monte Carlo simulations of diffusing STAT-3 molecules coupled with probabilistic modeling of dephosphorylation kinetics we found that predicted theoretical measures of STAT-3 diffusion likely overestimate the effective range of this signal. Compared to the inherently nucleus-directed movement of retrogradely transported signaling endosomes, diffusion of STAT-3 becomes less efficient at information transfer in spatial domains greater than 200 nanometers from the plasma membrane. Conclusion Our model suggests that cells might utilize two distinct information transmission paradigms: 1 fast local signaling via diffusion over spatial domains on the order of less than 200 nanometers; 2 long-distance signaling via information packets associated with the cytoskeletal transport apparatus. Our model supports previous observations suggesting that the signaling endosome hypothesis is a subset of a more general hypothesis that the most efficient mechanism for intracellular signaling-at-a-distance involves the association of signaling

  6. Decoupling internalization, acidification and phagosomal-endosomal/lysosomal fusion during phagocytosis of InlA coated beads in epithelial cells.

    Directory of Open Access Journals (Sweden)

    Craig D Blanchette

    Full Text Available BACKGROUND: Phagocytosis has been extensively examined in 'professional' phagocytic cells using pH sensitive dyes. However, in many of the previous studies, a separation between the end of internalization, beginning of acidification and completion of phagosomal-endosomal/lysosomal fusion was not clearly established. In addition, very little work has been done to systematically examine phagosomal maturation in 'non-professional' phagocytic cells. Therefore, in this study, we developed a simple method to measure and decouple particle internalization, phagosomal acidification and phagosomal-endosomal/lysosomal fusion in Madin-Darby Canine Kidney (MDCK and Caco-2 epithelial cells. METHODOLOGY/PRINCIPAL FINDINGS: Our method was developed using a pathogen mimetic system consisting of polystyrene beads coated with Internalin A (InlA, a membrane surface protein from Listeria monocytogenes known to trigger receptor-mediated phagocytosis. We were able to independently measure the rates of internalization, phagosomal acidification and phagosomal-endosomal/lysosomal fusion in epithelial cells by combining the InlA-coated beads (InlA-beads with antibody quenching, a pH sensitive dye and an endosomal/lysosomal dye. By performing these independent measurements under identical experimental conditions, we were able to decouple the three processes and establish time scales for each. In a separate set of experiments, we exploited the phagosomal acidification process to demonstrate an additional, real-time method for tracking bead binding, internalization and phagosomal acidification. CONCLUSIONS/SIGNIFICANCE: Using this method, we found that the time scales for internalization, phagosomal acidification and phagosomal-endosomal/lysosomal fusion ranged from 23-32 min, 3-4 min and 74-120 min, respectively, for MDCK and Caco-2 epithelial cells. Both the static and real-time methods developed here are expected to be readily and broadly applicable, as they simply

  7. Actin polymerization in the endosomal pathway, but not on the Coxiella-containing vacuole, is essential for pathogen growth.

    Directory of Open Access Journals (Sweden)

    Heather E Miller

    2018-04-01

    Full Text Available Coxiella burnetii is an intracellular bacterium that replicates within an expansive phagolysosome-like vacuole. Fusion between the Coxiella-containing vacuole (CCV and late endosomes/multivesicular bodies requires Rab7, the HOPS tethering complex, and SNARE proteins, with actin also speculated to play a role. Here, we investigated the importance of actin in CCV fusion. Filamentous actin patches formed around the CCV membrane that were preferred sites of vesicular fusion. Accordingly, the mediators of endolysosomal fusion Rab7, VAMP7, and syntaxin 8 were concentrated in CCV actin patches. Generation of actin patches required C. burnetii type 4B secretion and host retromer function. Patches decorated with VPS29 and VPS35, components of the retromer, FAM21 and WASH, members of the WASH complex that engage the retromer, and Arp3, a component of the Arp2/3 complex that generates branched actin filaments. Depletion by siRNA of VPS35 or VPS29 reduced CCV actin patches and caused Rab7 to uniformly distribute in the CCV membrane. C. burnetii grew normally in VPS35 or VPS29 depleted cells, as well as WASH-knockout mouse embryo fibroblasts, where CCVs are devoid of actin patches. Endosome recycling to the plasma membrane and trans-Golgi of glucose transporter 1 (GLUT1 and cationic-independent mannose-6-phosphate receptor (CI-M6PR, respectively, was normal in infected cells. However, siRNA knockdown of retromer resulted in aberrant trafficking of GLUT1, but not CI-M6PR, suggesting canonical retrograde trafficking is unaffected by retromer disruption. Treatment with the specific Arp2/3 inhibitor CK-666 strongly inhibited CCV formation, an effect associated with altered endosomal trafficking of transferrin receptor. Collectively, our results show that CCV actin patches generated by retromer, WASH, and Arp2/3 are dispensable for CCV biogenesis and stability. However, Arp2/3-mediated production of actin filaments required for cargo transport within the

  8. The atrial fibrillation ablation pilot study

    DEFF Research Database (Denmark)

    Arbelo, Elena; Brugada, Josep; Hindricks, Gerhard

    2014-01-01

    AIMS: The Atrial Fibrillation Ablation Pilot Study is a prospective registry designed to describe the clinical epidemiology of patients undergoing an atrial fibrillation (AFib) ablation, and the diagnostic/therapeutic processes applied across Europe. The aims of the 1-year follow-up were to analyse...... was achieved in 40.7% of patients (43.7% in paroxysmal AF; 30.2% in persistent AF; 36.7% in long-lasting persistent AF). A second ablation was required in 18% of the cases and 43.4% were under antiarrhythmic treatment. Thirty-three patients (2.5%) suffered an adverse event, 272 (21%) experienced a left atrial...... tachycardia, and 4 patients died (1 haemorrhagic stroke, 1 ventricular fibrillation in a patient with ischaemic heart disease, 1 cancer, and 1 of unknown cause). CONCLUSION: The AFib Ablation Pilot Study provided crucial information on the epidemiology, management, and outcomes of catheter ablation of AFib...

  9. Advances in laser ablation of materials

    International Nuclear Information System (INIS)

    Singh, R.K.; Lowndes, D.H.; Chrisey, D.B.; Fogarassy, E.; Narayan, J.

    1998-01-01

    The symposium, Advances in Laser Ablation of Materials, was held at the 1998 MRS Spring Meeting in San Francisco, California. The papers in this symposium illustrate the advances in pulsed laser ablation for a wide variety of applications involving semiconductors, superconductors, metals, ceramics, and polymers. In particular, advances in the deposition of oxides and related materials are featured. Papers dealing with both fundamentals and the applications of laser ablation are presented. Topical areas include: fundamentals of ablation and growth; in situ diagnostics and nanoscale synthesis advances in laser ablation techniques; laser surface processing; pulsed laser deposition of ferroelectric, magnetic, superconducting and optoelectronic thin films; and pulsed laser deposition of carbon-based and polymeric materials. Sixty papers have been processed separately for inclusion on the data base

  10. Growth of anatase and rutile phase TiO{sub 2} nanoparticles using pulsed laser ablation in liquid: Influence of surfactant addition and ablation time variation

    Energy Technology Data Exchange (ETDEWEB)

    Chaturvedi, Amita, E-mail: amita-chaturvedi@rrcat.gov.in [Laser Material Processing Division, Raja Ramanna Centre for Advanced Technology, Indore 452013, MP (India); Joshi, M.P. [Laser Material Processing Division, Raja Ramanna Centre for Advanced Technology, Indore 452013, MP (India); Homi Bhabha National Institute, Training School Complex, Anushakti Nagar, Mumbai – 400094 (India); Mondal, P.; Sinha, A.K.; Srivastava, A.K. [Indus Synchrotron Utilization Division, Raja Ramanna Centre for Advanced Technology, Indore 452013, MP (India)

    2017-02-28

    Highlights: • Ablations of Ti metal target were carried out in DI water and in 0.001 M SDS solution for different times using PLAL process. • Different characterization studies have been carried out to confirm the growth of TiO{sub 2} nanoparticles in both the liquid mediums. • Anatase phase TiO{sub 2} nanoparticles were obtained in DI water and rutile phase in 0.001 M SDS aqueous solution. • In surfactant solution, longer time ablation leads depletion of SDS molecules causes growth of anatase phase for 90 min. • Our studies confirmed the role of liquid ambience conditions variation over the different phase formations of nanoparticles. - Abstract: Titanium dioxide (TiO{sub 2}) nanoparticles were grown using nanosecond pulsed laser ablation of Ti target in DI water and in 0.001 M sodium dodecyl sulfate (SDS) surfactant aqueous solution. Growth was carried out with varying ablation times i. e. 30 min, 60 min and 90 min. The objective of our study was to investigate the influence of variations in liquid ambience conditions on the growth of the nanoparticles in a pulsed laser ablation in liquid (PLAL) process. Size, composition and optical properties of the grown TiO{sub 2} nanoparticles were investigated using transmission electron microscopy (TEM), X-ray photoelectron spectroscopy (XPS), optical absorption, photoluminescence (PL) spectroscopy and X-ray diffraction (XRD) studies. The obtained nanoparticles of TiO{sub 2} were found almost spherical in shape and polycrystalline in nature in both the liquid mediums i.e. DI water and aqueous solution of surfactant. Nanoparticles number density was also found to increase with increasing ablation time in both the liquid mediums. However crystalline phase of the grown TiO{sub 2} nanoparticles differs with the change in liquid ambience conditions. Selected area electron diffraction (SAED), PL and XRD studies suggest that DI water ambience is favorable for the growth of anatase phase TiO{sub 2} nanoparticles for all

  11. Catheter Ablation versus Thoracoscopic Surgical Ablation in Long Standing Persistent Atrial Fibrillation (CASA-AF): study protocol for a randomised controlled trial.

    Science.gov (United States)

    Khan, Habib Rehman; Kralj-Hans, Ines; Haldar, Shouvik; Bahrami, Toufan; Clague, Jonathan; De Souza, Anthony; Francis, Darrel; Hussain, Wajid; Jarman, Julian; Jones, David Gareth; Mediratta, Neeraj; Mohiaddin, Raad; Salukhe, Tushar; Jones, Simon; Lord, Joanne; Murphy, Caroline; Kelly, Joanna; Markides, Vias; Gupta, Dhiraj; Wong, Tom

    2018-02-20

    Atrial fibrillation is the commonest arrhythmia which raises the risk of heart failure, thromboembolic stroke, morbidity and death. Pharmacological treatments of this condition are focused on heart rate control, rhythm control and reduction in risk of stroke. Selective ablation of cardiac tissues resulting in isolation of areas causing atrial fibrillation is another treatment strategy which can be delivered by two minimally invasive interventions: percutaneous catheter ablation and thoracoscopic surgical ablation. The main purpose of this trial is to compare the effectiveness and safety of these two interventions. Catheter Ablation versus Thoracoscopic Surgical Ablation in Long Standing Persistent Atrial Fibrillation (CASA-AF) is a prospective, multi-centre, randomised controlled trial within three NHS tertiary cardiovascular centres specialising in treatment of atrial fibrillation. Eligible adults (n = 120) with symptomatic, long-standing, persistent atrial fibrillation will be randomly allocated to either catheter ablation or thoracoscopic ablation in a 1:1 ratio. Pre-determined lesion sets will be delivered in each treatment arm with confirmation of appropriate conduction block. All patients will have an implantable loop recorder (ILR) inserted subcutaneously immediately following ablation to enable continuous heart rhythm monitoring for at least 12 months. The devices will be programmed to detect episodes of atrial fibrillation and atrial tachycardia ≥ 30 s in duration. The patients will be followed for 12 months, completing appropriate clinical assessments and questionnaires every 3 months. The ILR data will be wirelessly transmitted daily and evaluated every month for the duration of the follow-up. The primary endpoint in the study is freedom from atrial fibrillation and atrial tachycardia at the end of the follow-up period. The CASA-AF Trial is a National Institute for Health Research-funded study that will provide first-class evidence on the

  12. Catheter ablation of epicardial ventricular tachycardia

    Directory of Open Access Journals (Sweden)

    Takumi Yamada, MD, PhD

    2014-08-01

    Full Text Available Ventricular tachycardias (VTs can usually be treated by endocardial catheter ablation. However, some VTs can arise from the epicardial surface, and their substrate can be altered only by epicardial catheter ablation. There are two approaches to epicardial catheter ablation: transvenous and transthoracic. The transvenous approach through the coronary venous system (CVS has been commonly used because it is easily accessible. However, this approach may be limited by the distribution of the CVS and insufficient radiofrequency energy delivery. Transthoracic epicardial catheter ablation has been developed to overcome these limitations of the transvenous approach. It is a useful supplemental or even preferred strategy to eliminate epicardial VTs in the electrophysiology laboratory. This technique has been applied for scar-related VTs secondary to often non-ischemic cardiomyopathy and sometimes ischemic cardiomyopathy, and idiopathic VTs as the epicardial substrates of these VTs have become increasingly recognized. When endocardial ablation and epicardial ablation through the CVS are unsuccessful, transthoracic epicardial ablation should be the next option. Intrapericardial access is usually obtained through a subxiphoidal pericardial puncture. This approach might not be possible in patients with pericardial adhesions caused by prior cardiac surgery or pericarditis. In such cases, a hybrid procedure involving surgical access with a subxiphoid pericardial window and a limited anterior or lateral thoracotomy might be a feasible and safe method of performing an epicardial catheter ablation in the electrophysiology laboratory. Potential complications associated with this technique include bleeding and collateral damage to the coronary arteries and phrenic nerve. Although the risk of these complications is low, electrophysiologists who attempt epicardial catheter ablation should know the complications associated with this technique, how to minimize their

  13. Determination of ablation threshold for composite resins and amalgam irradiated with femtosecond laser pulses

    International Nuclear Information System (INIS)

    Freitas, A Z; Samad, R E; Zezell, D M; Vieira Jr, N D; Freschi, L R; Gouw-Soares, S C

    2010-01-01

    The use of laser for caries removal and cavity preparation is already a reality in the dental clinic. The objective of the present study was to consider the viability of ultrashort laser pulses for restorative material selective removal, by determining the ablation threshold fluence for composite resins and amalgam irradiated with femtosecond laser pulses. Lasers pulses centered at 830 nm with 50 fs of duration and 1 kHz of repetition rate, with energies in the range of 300 to 770 μJ were used to irradiate the samples. The samples were irradiated using two different geometrical methods for ablation threshold fluence determinations and the volume ablation was measured by optical coherence tomography. The shape of the ablated surfaces were analyzed by optical microscopy and scanning electron microscopy. The determined ablation threshold fluence is 0.35 J/cm 2 for the composite resins Z-100 and Z-350, and 0.25 J/cm 2 for the amalgam. These values are half of the value for enamel in this temporal regime. Thermal damages were not observed in the samples. Using the OCT technique (optical coherence tomography) was possible to determine the ablated volume and the total mass removed

  14. Decoupling Internalization, Acidification and Phagosmal-Endosomal/Iysosomal Phagocytosis of Internalin A coated Beads in epithelial cells

    Energy Technology Data Exchange (ETDEWEB)

    Blanchette, C D; Woo, Y; Thomas, C; Shen, N; Sulchek, T A; Hiddessen, A L

    2008-12-22

    Phagocytosis has been extensively examined in 'professional' phagocytic cells using pH sensitive dyes. However, in many of the previous studies, a separation between the end of internalization, beginning of acidification and completion of phagosomal-endosomal/lysosomal fusion was not clearly established, and in several cases, it was treated as a one-step process. In addition, very little work has been done to systematically examine phagosomal maturation in 'non-professional' phagocytic cells, such as epithelial cells. Therefore, in this study, we developed a simple and novel method to decouple and accurately measure particle internalization, phagosomal acidification and phagosomal-endosomal/lysosomal fusion in Madin-Darby Canine Kidney (MDCK) and Caco-2 epithelial cells. Our method was developed using a pathogen mimetic system consisting of polystyrene beads coated with Internalin A (InlA), a membrane surface protein from Listeria monocytogenes known to trigger receptor-mediated internalization. We achieved independent measurements of the rates of internalization, phagosomal acidification and phagosomal-endosomal/lysosomal fusion in epithelial cells by combining the InlA-coated beads (InlA-beads) with antibody quenching, pH sensitive dyes and endosomal/lysosomal dyes, as follows: the rate of InlA bead internalization was measured via antibody quenching of a pH independent dye (Alexa488) conjugated to InlA-beads, the rate at which phagosomes containing internalized InlA beads became acidified was measured using a pH dependent dye (FITC) conjugated to the beads and the rate of phagosomal-endosomal/lysosomal fusion was measured using a combination of unlabeled InlA-beads and an endosomal/lysosomal dye. By performing these independent measurements under identical experimental conditions, we were able to decouple the three processes and establish time scales for each. In a separate set of experiments, we also exploited the phagosomal acidification

  15. Optical properties of tin oxide nanoparticles prepared by laser ablation in water: Influence of laser ablation time duration and laser fluence

    International Nuclear Information System (INIS)

    Desarkar, Himadri Sankar; Kumbhakar, P.; Mitra, A.K.

    2012-01-01

    Colloidal tin oxide nanoparticles are prepared by laser (having a wavelength of 1064 nm) ablation of tin metallic target immersed in pure deionized water. The influences of laser ablation time and laser fluence on the size and optical properties of the synthesized nanoparticles are studied. Prepared tin oxide nanoparticles are characterized by transmission electron microscope, selected area electron diffraction and UV–Visible absorption spectroscopy. The morphology of prepared tin oxide nanoparticles is found to be mostly spherical and with sizes in the nanometric range (mean radius of 3.2 to 7.3 nm). The measured UV–Visible absorption spectra show the presence of absorption peaks in the ultraviolet region. The band gap energy of samples prepared with different laser ablation time duration is calculated and is found to be increased with decrease in size (radius) of the prepared nanoparticles. Photoluminescence emission measurements at room temperature show that all the samples exhibit photoluminescence in the visible region. The peak photoluminescence emission intensity in the sample prepared with 50 min of laser ablation time is 3.5 times larger than that obtained in the sample prepared with 10 min of laser ablation time. - Highlights: ► SnO 2 nanoparticles (6.4–14.6 nm) are prepared by laser ablation in liquid technique. ► The influences of laser ablation time and laser fluence are studied. ► Samples are characterized by TEM and UV–Visible absorption spectroscopy. ► UV–Visible absorption spectra exhibit quantum confinement effect. ► Samples exhibit enhanced photoluminescence emissions in the visible region.

  16. Physical mechanisms of SiNx layer structuring with ultrafast lasers by direct and confined laser ablation

    International Nuclear Information System (INIS)

    Rapp, S.; Heinrich, G.; Wollgarten, M.; Huber, H. P.; Schmidt, M.

    2015-01-01

    In the production process of silicon microelectronic devices and high efficiency silicon solar cells, local contact openings in thin dielectric layers are required. Instead of photolithography, these openings can be selectively structured with ultra-short laser pulses by confined laser ablation in a fast and efficient lift off production step. Thereby, the ultrafast laser pulse is transmitted by the dielectric layer and absorbed at the substrate surface leading to a selective layer removal in the nanosecond time domain. Thermal damage in the substrate due to absorption is an unwanted side effect. The aim of this work is to obtain a deeper understanding of the physical laser-material interaction with the goal of finding a damage-free ablation mechanism. For this, thin silicon nitride (SiN x ) layers on planar silicon (Si) wafers are processed with infrared fs-laser pulses. Two ablation types can be distinguished: The known confined ablation at fluences below 300 mJ/cm 2 and a combined partial confined and partial direct ablation at higher fluences. The partial direct ablation process is caused by nonlinear absorption in the SiN x layer in the center of the applied Gaussian shaped laser pulses. Pump-probe investigations of the central area show ultra-fast reflectivity changes typical for direct laser ablation. Transmission electron microscopy results demonstrate that the Si surface under the remaining SiN x island is not damaged by the laser ablation process. At optimized process parameters, the method of direct laser ablation could be a good candidate for damage-free selective structuring of dielectric layers on absorbing substrates

  17. Benign thyroid nodule unresponsive to radiofrequency ablation treated with laser ablation: a case report.

    Science.gov (United States)

    Oddo, Silvia; Balestra, Margherita; Vera, Lara; Giusti, Massimo

    2018-05-11

    Radiofrequency ablation and laser ablation are safe and effective techniques for reducing thyroid nodule volume, neck symptoms, and cosmetic complaints. Therapeutic success is defined as a nodule reduction > 50% between 6 and 12 months after the procedure, but a percentage of nodules inexplicably do not respond to thermal ablation. We describe the case of a young Caucasian woman with a solid benign thyroid nodule who refused surgery and who had undergone radiofrequency ablation in 2013. The nodule did not respond in terms of either volume reduction or improvement in neck symptoms. After 2 years, given the patient's continued refusal of thyroidectomy, we proposed laser ablation. The nodule displayed a significant volume reduction (- 50% from radiofrequency ablation baseline volume, - 57% from laser ablation baseline), and the patient reported a significant improvement in neck symptoms (from 6/10 to 1/10 on a visual analogue scale). We conjecture that some benign thyroid nodules may be intrinsically resistant to necrosis when one specific ablation technique is used, but may respond to another technique. To the best of our knowledge, this is the first description of the effect of performing a different percutaneous ablation technique in a nodule that does not respond to radiofrequency ablation.

  18. The Serotonin Transporter Undergoes Constitutive Internalization and Is Primarily Sorted to Late Endosomes and Lysosomal Degradation*

    Science.gov (United States)

    Rahbek-Clemmensen, Troels; Bay, Tina; Eriksen, Jacob; Gether, Ulrik; Jørgensen, Trine Nygaard

    2014-01-01

    The serotonin transporter (SERT) plays a critical role in regulating serotonin signaling by mediating reuptake of serotonin from the extracellular space. The molecular and cellular mechanisms controlling SERT levels in the membrane remain poorly understood. To study trafficking of the surface resident SERT, two functional epitope-tagged variants were generated. Fusion of a FLAG-tagged one-transmembrane segment protein Tac to the SERT N terminus generated a transporter with an extracellular epitope suited for trafficking studies (TacSERT). Likewise, a construct with an extracellular antibody epitope was generated by introducing an HA (hemagglutinin) tag in the extracellular loop 2 of SERT (HA-SERT). By using TacSERT and HA-SERT in antibody-based internalization assays, we show that SERT undergoes constitutive internalization in a dynamin-dependent manner. Confocal images of constitutively internalized SERT demonstrated that SERT primarily co-localized with the late endosomal/lysosomal marker Rab7, whereas little co-localization was observed with the Rab11, a marker of the “long loop” recycling pathway. This sorting pattern was distinct from that of a prototypical recycling membrane protein, the β2-adrenergic receptor. Furthermore, internalized SERT co-localized with the lysosomal marker LysoTracker and not with transferrin. The sorting pattern was further confirmed by visualizing internalization of SERT using the fluorescent cocaine analog JHC1-64 and by reversible and pulse-chase biotinylation assays showing evidence for lysosomal degradation of the internalized transporter. Finally, we found that SERT internalized in response to stimulation with 12-myristate 13-acetate co-localized primarily with Rab7- and LysoTracker-positive compartments. We conclude that SERT is constitutively internalized and that the internalized transporter is sorted mainly to degradation. PMID:24973209

  19. Eps homology domain endosomal transport proteins differentially localize to the neuromuscular junction

    Directory of Open Access Journals (Sweden)

    Mate Suzanne E

    2012-09-01

    Full Text Available Abstract Background Recycling of endosomes is important for trafficking and maintenance of proteins at the neuromuscular junction (NMJ. We have previously shown high expression of the endocytic recycling regulator Eps15 homology domain-containing (EHD1 proteinin the Torpedo californica electric organ, a model tissue for investigating a cholinergic synapse. In this study, we investigated the localization of EHD1 and its paralogs EHD2, EHD3, and EHD4 in mouse skeletal muscle, and assessed the morphological changes in EHD1−/− NMJs. Methods Localization of the candidate NMJ protein EHD1 was assessed by confocal microscopy analysis of whole-mount mouse skeletal muscle fibers after direct gene transfer and immunolabeling. The potential function of EHD1 was assessed by specific force measurement and α-bungarotoxin-based endplate morphology mapping in EHD1−/− mouse skeletal muscle. Results Endogenous EHD1 localized to primary synaptic clefts of murine NMJ, and this localization was confirmed by expression of recombinant green fluorescent protein labeled-EHD1 in murine skeletal muscle in vivo. EHD1−/− mouse skeletal muscle had normal histology and NMJ morphology, and normal specific force generation during muscle contraction. The EHD 1–4 proteins showed differential localization in skeletal muscle: EHD2 to muscle vasculature, EHD3 to perisynaptic regions, and EHD4 to perinuclear regions and to primary synaptic clefts, but at lower levels than EHD1. Additionally, specific antibodies raised against mammalian EHD1-4 recognized proteins of the expected mass in the T. californica electric organ. Finally, we found that EHD4 expression was more abundant in EHD1−/− mouse skeletal muscle than in wild-type skeletal muscle. Conclusion EHD1 and EHD4 localize to the primary synaptic clefts of the NMJ. Lack of obvious defects in NMJ structure and muscle function in EHD1−/− muscle may be due to functional compensation by other EHD paralogs.

  20. Radiofrequency ablation of lung and liver lesions using CT fluoroscopy

    International Nuclear Information System (INIS)

    Chai, A.; Glenn, D.

    2002-01-01

    Full text: Tumour ablation with radiofrequency (RF) energy is a relatively new procedure for the treatment of focal malignant disease. At our institution this is currently being used in the treatment of certain liver and lung lesions with the patients involved being enrolled in clinical trials. The poster describes the technique used at our institution for the placement of the radiofrequency ablation electrode using CT fluoroscopy. Criteria for patient selection are included. Complications from the procedure are described, as well as follow up appearances and results. Our results from the treatment of primary and secondary lesions in the liver correlate well with published literature. Treatment is still not as successful as surgical resection but there is significantly less morbidity. Where this method may be appropriate is when the patient is not a candidate for surgical resection. The treatment of colorectal metastases in the lung shows early promise as a possible second line treatment (as for liver) where the patient is not a candidate for surgery. Preliminary results are soon to be published in conjunction with the Department of Surgery at our institution. RF Electrode placement using CT Fluoroscopy is performed at our institution. While still at its early stages, RF Ablation shows promise as a possible second line treatment (with other adjuvant therapy) for the management of focal malignant disease in the lung and liver. Copyright (2002) Blackwell Science Pty Ltd

  1. Transarterial ablation of hepatocellular carcinoma. Status and developments

    International Nuclear Information System (INIS)

    Radeleff, B.A.; Stampfl, U.; Sommer, C.M.; Bellemann, N.; Kauczor, H.U.; Hoffmann, K.; Ganten, T.; Ehehalt, R.

    2012-01-01

    Hepatocellular carcinoma (HCC) is the fifth most common cancer worldwide and represents the main cause of death among European patients with liver cirrhosis. Only 30-40% of patients diagnosed with HCC are candidates for curative treatment options (e.g. surgical resection, liver transplantation or ablation). The remaining majority of patients must undergo local regional and palliative therapies. Transvascular ablation of HCC takes advantage of the fact that the hypervascularized HCC receives most of its blood supply from the hepatic artery. In this context transvascular ablation describes different therapy regimens which can be assigned to four groups: cTACE (conventional transarterial chemoembolization), bland embolization (transarterial embolization TAE), DEB-TACE (TACE with drug-eluting beads, DEB) and SIRT (selective internal radiation therapy, radioembolization). Conventional TACE is the most common type of transvascular ablation and represents a combination of intra-arterial chemotherapy and embolization with occlusion of the arterial blood supply. However, there is no standardized regimen with respect to the chemotherapeutic drug, the embolic agent, the usage of lipiodol and the interval between the TACE procedures. Even the exact course of a cTACE procedure (order of chemotherapy or embolization) is not standardized. It remains unclear whether or not intra-arterial chemotherapy is definitely required as bland embolization using very small, tightly calibrated spherical particles (without intra-arterial administration of a chemotherapeutic drug) shows tumor necrosis comparable to cTACE. For DEB-TACE microparticles loaded with a chemotherapeutic drug combine the advantages of cTACE and bland embolization. Thereby, a continuing chemotherapeutic effect within the tumor might cause a further increase in intratumoral cytotoxicity and at the same time a decrease in systemic toxicity. (orig.) [de

  2. A spectral analysis of ablating meteors

    Science.gov (United States)

    Bloxam, K.; Campbell-Brown, M.

    2017-09-01

    Meteor ablation features in the spectral lines occurring at 394, 436, 520, and 589 nm were observed using a four-camera spectral system between September and December 2015. In conjunction with this multi-camera system the Canadian Automated Meteor Observatory was used to observe the orbital parameters and fragmentation of these meteors. In total, 95 light curves with complete data in the 520 and 589 nm filters were analyzed; some also had partial or complete data in the 394 nm filter, but no usable data was collected with the 436 nm filter. Of the 95 events, 70 exhibited some degree of differential ablation, and in all except 3 of these 70 events the 589 nm filter started or ended sooner compared with the 520 nm filter, indicating early ablation at the 589 nm wavelength. In the majority of cases the meteor showed evidence of fragmentation regardless of the type of ablation (differential or uniform). A surprising result was the lack of correlation found concerning the KB parameter, linked to meteoroid strength, and differential ablation. In addition, 22 shower-associated meteors were observed; Geminids showed mainly slight differential ablation, while Taurids were more likely to ablate uniformly.

  3. Early to Late Endosome Trafficking Controls Secretion and Zymogen Activation in Rodent and Human Pancreatic Acinar CellsSummary

    Directory of Open Access Journals (Sweden)

    Scott W. Messenger

    2015-11-01

    Full Text Available Background & Aims: Pancreatic acinar cells have an expanded apical endosomal system, the physiologic and pathophysiologic significance of which is still emerging. Phosphatidylinositol-3,5-bisphosphate [PI(3,5P2] is an essential phospholipid generated by phosphatidylinositol 3-phosphate 5-kinase (PIKfyve, which phosphorylates phosphatidylinositol-3-phosphate (PI3P. PI(3,5P2 is necessary for maturation of early endosomes (EE to late endosomes (LE. Inhibition of EE to LE trafficking enhances anterograde endosomal trafficking and secretion at the plasma membrane by default through a recycling endosome (RE intermediate. We assessed the effects of modulating PIKfyve activity on apical trafficking and pancreatitis responses in pancreatic acinar cells. Methods: Inhibition of EE to LE trafficking was achieved using pharmacologic inhibitors of PIKfyve, expression of dominant negative PIKfyve K1877E, or constitutively active Rab5-GTP Q79L. Anterograde endosomal trafficking was manipulated by expression of constitutively active and dominant negative Rab11a mutants. The effects of these agents on secretion, endolysosomal exocytosis of lysosome associated membrane protein (LAMP1, and trypsinogen activation in response to supramaximal cholecystokinin (CCK-8, bile acids, and cigarette toxin was determined. Results: PIKfyve inhibition increased basal and stimulated secretion. Adenoviral overexpression of PIKfyve decreased secretion leading to cellular death. Expression of Rab5-GTP Q79L or Rab11a-GTP Q70L enhanced secretion. Conversely, dominant-negative Rab11a-GDP S25N reduced secretion. High-dose CCK inhibited endolysosomal exocytosis that was reversed by PIKfyve inhibition. PIKfyve inhibition blocked intracellular trypsin accumulation and cellular damage responses to supramaximal CCK-8, tobacco toxin, and bile salts in both rodent and human acini. Conclusions: These data demonstrate that EE-LE trafficking acutely controls acinar secretion and the intracellular

  4. An experimental study of simultaneous ablation with dual probes in radiofrequency thermal ablation

    International Nuclear Information System (INIS)

    Jang, Il Soo; Rhim, Hyun Chul; Koh, Byung Hee; Cho, On Koo; Seo, Heung Suk; Kim, Yong Soo; Kim, Young Sun; Heo, Jeong Nam

    2003-01-01

    To determine the differences between sequential ablation with a single probe and simultaneous ablation with dual probes. Using two 14-gauge expandable probes (nine internal prongs with 4-cm deployment), radiofrequency was applied sequentially (n=8) or simultaneously (n=8) to ten ex-vivo cow livers. Before starting ablation, two RF probes with an inter-probe space of 2 cm (n=8) or 3 cm (n=8) were inserted. In the sequential group, switching the connecting cable to an RF generator permitted ablation with the second probe just after ablation with the first probe had finished. In the simultaneous group, single ablation was performed only after connecting the shafts of both RF probes using a connection device. Each ablation lasted 7 minutes at a target temperature of 105-110 .deg. C. The size and shape of the ablated area, and total ablation time were then compared between the two groups. With 2-cm spacing, the group, mean length and overlapping width of ablated lesions were, respectively, 5.20 and 5.05 cm in the sequential group (n=4), and 5.81 and 5.65 cm in the simultaneous group (n=4). With 3-cm spacing, the corresponding figures were 4.99 and 5.60 cm in the sequential group (n=4), and 6.04 and 6.78 cm in the simultaneous group (n=4). With 2-cm spacing, the mean depth of the proximal waist was 0.58 cm in the sequential (group and 0.28 cm in the simultaneous group, while with 3-cm spacing, the corresponding figures were 1.65 and 1.48 cm. In neither group was there a distal waist. Mean total ablation time was 23.4 minutes in the sequential group and 14 minutes in the simultaneous group. In terms of ablation size and ablation time, simultaneous radiofrequency ablation with dual probes is superior to sequential ablation with a single probe. A simultaneous approach will enable an operator to overcome difficulty in probe repositioning during overlapping ablation, resulting in complete ablation with a successful safety margin

  5. Chemically assisted laser ablation ICP mass spectrometry.

    Science.gov (United States)

    Hirata, Takafumi

    2003-01-15

    A new laser ablation technique combined with a chemical evaporation reaction has been developed for elemental ratio analysis of solid samples using an inductively coupled plasma mass spectrometer (ICPMS). Using a chemically assisted laser ablation (CIA) technique developed in this study, analytical repeatability of the elemental ratio measurement was successively improved. To evaluate the reliability of the CLA-ICPMS technique, Pb/U isotopic ratios were determined for zircon samples that have previously been analyzed by other techniques. Conventional laser ablation for Pb/U shows a serious elemental fractionation during ablation mainly due to the large difference in elemental volatility between Pb and U. In the case of Pb/U ratio measurement, a Freon R-134a gas (1,1,1,2-tetrafluoroethane) was introduced into the laser cell as a fluorination reactant. The Freon gas introduced into the laser cell reacts with the ablated sample U, and refractory U compounds are converted to a volatile U fluoride compound (UF6) under the high-temperature condition at the ablation site. This avoids the redeposition of U around the ablation pits. Although not all the U is reacted with Freon, formation of volatile UF compounds improves the transmission efficiency of U. Typical precision of the 206Pb/238U ratio measurement is 3-5% (2sigma) for NIST SRM 610 and Nancy 91500 zircon standard, and the U-Pb age data obtained here show good agreement within analytical uncertainties with the previously reported values. Since the observed Pb/U ratio for solid samples is relatively insensitive to laser power and ablation time, optimization of ablation conditions or acquisition parameters no longer needs to be performed on a sample-to-sample basis.

  6. Direct His bundle pacing post AVN ablation.

    Science.gov (United States)

    Lakshmanadoss, Umashankar; Aggarwal, Ashim; Huang, David T; Daubert, James P; Shah, Abrar

    2009-08-01

    Atrioventricular nodal (AVN) ablation with concomitant pacemaker implantation is one of the strategies that reduce symptoms in patients with atrial fibrillation (AF). However, the long-term adverse effects of right ventricular (RV) apical pacing have led to the search for alternating sites of pacing. Biventricular pacing produces a significant improvement in functional capacity over RV pacing in patients undergoing AVN ablation. Another alternative site for pacing is direct His bundle to reduce the adverse outcome of RV pacing. Here, we present a case of direct His bundle pacing using steerable lead delivery system in a patient with symptomatic paroxysmal AF with concurrent AVN ablation.

  7. TEM investigations of laser ablated particles

    International Nuclear Information System (INIS)

    Fliegel, D.; Dundas, S.; Kosler, J.; Klementova, M.

    2009-01-01

    Full text: Laser ablation inductively coupled plasma mass spectrometry suffers from fractionation effects hindering a non matrix matched calibration strategy. Different reasons for elemental fractionation that are related to the laser ablation, the transport and the vaporization in the plasma are discussed. One major question to be addressed linked to the vaporization yield in the ICP is in which of mineralogical phase the different ablated particle sizes enter the plasma. This contribution will investigate particles generated by a 213 nm laser from different samples such as minerals and alloys with respect to their chemical and phase compositions using high resolution TEM. (author)

  8. Investigation of different liquid media and ablation times on pulsed laser ablation synthesis of aluminum nanoparticles

    International Nuclear Information System (INIS)

    Baladi, Arash; Sarraf Mamoory, Rasoul

    2010-01-01

    Aluminum nanoparticles were synthesized by pulsed laser ablation of Al targets in ethanol, acetone, and ethylene glycol. Transmission Electron Microscope (TEM) and Scanning Electron Microscope (SEM) images, Particle size distribution diagram from Laser Particle Size Analyzer (LPSA), UV-visible absorption spectra, and weight changes of targets were used for the characterization and comparison of products. The experiments demonstrated that ablation efficiency in ethylene glycol is too low, in ethanol is higher, and in acetone is highest. Comparison between ethanol and acetone clarified that acetone medium leads to finer nanoparticles (mean diameter of 30 nm) with narrower size distribution (from 10 to 100 nm). However, thin carbon layer coats some of them, which was not observed in ethanol medium. It was also revealed that higher ablation time resulted in higher ablated mass, but lower ablation rate. Finer nanoparticles, moreover, were synthesized in higher ablation times.

  9. Investigation of different liquid media and ablation times on pulsed laser ablation synthesis of aluminum nanoparticles

    Energy Technology Data Exchange (ETDEWEB)

    Baladi, Arash [Materials Engineering Department, Tarbiat Modares University, Jalal Al Ahmad, P.O. Box 14115-143, Tehran (Iran, Islamic Republic of); Sarraf Mamoory, Rasoul, E-mail: rsarrafm@modares.ac.ir [Materials Engineering Department, Tarbiat Modares University, Jalal Al Ahmad, P.O. Box 14115-143, Tehran (Iran, Islamic Republic of)

    2010-10-01

    Aluminum nanoparticles were synthesized by pulsed laser ablation of Al targets in ethanol, acetone, and ethylene glycol. Transmission Electron Microscope (TEM) and Scanning Electron Microscope (SEM) images, Particle size distribution diagram from Laser Particle Size Analyzer (LPSA), UV-visible absorption spectra, and weight changes of targets were used for the characterization and comparison of products. The experiments demonstrated that ablation efficiency in ethylene glycol is too low, in ethanol is higher, and in acetone is highest. Comparison between ethanol and acetone clarified that acetone medium leads to finer nanoparticles (mean diameter of 30 nm) with narrower size distribution (from 10 to 100 nm). However, thin carbon layer coats some of them, which was not observed in ethanol medium. It was also revealed that higher ablation time resulted in higher ablated mass, but lower ablation rate. Finer nanoparticles, moreover, were synthesized in higher ablation times.

  10. Lsd1 Ablation Triggers Metabolic Reprogramming of Brown Adipose Tissue

    Directory of Open Access Journals (Sweden)

    Delphine Duteil

    2016-10-01

    Full Text Available Previous work indicated that lysine-specific demethylase 1 (Lsd1 can positively regulate the oxidative and thermogenic capacities of white and beige adipocytes. Here we investigate the role of Lsd1 in brown adipose tissue (BAT and find that BAT-selective Lsd1 ablation induces a shift from oxidative to glycolytic metabolism. This shift is associated with downregulation of BAT-specific and upregulation of white adipose tissue (WAT-selective gene expression. This results in the accumulation of di- and triacylglycerides and culminates in a profound whitening of BAT in aged Lsd1-deficient mice. Further studies show that Lsd1 maintains BAT properties via a dual role. It activates BAT-selective gene expression in concert with the transcription factor Nrf1 and represses WAT-selective genes through recruitment of the CoREST complex. In conclusion, our data uncover Lsd1 as a key regulator of gene expression and metabolic function in BAT.

  11. Voltage and pace-capture mapping of linear ablation lesions overestimates chronic ablation gap size.

    Science.gov (United States)

    O'Neill, Louisa; Harrison, James; Chubb, Henry; Whitaker, John; Mukherjee, Rahul K; Bloch, Lars Ølgaard; Andersen, Niels Peter; Dam, Høgni; Jensen, Henrik K; Niederer, Steven; Wright, Matthew; O'Neill, Mark; Williams, Steven E

    2018-04-26

    Conducting gaps in lesion sets are a major reason for failure of ablation procedures. Voltage mapping and pace-capture have been proposed for intra-procedural identification of gaps. We aimed to compare gap size measured acutely and chronically post-ablation to macroscopic gap size in a porcine model. Intercaval linear ablation was performed in eight Göttingen minipigs with a deliberate gap of ∼5 mm left in the ablation line. Gap size was measured by interpolating ablation contact force values between ablation tags and thresholding at a low force cut-off of 5 g. Bipolar voltage mapping and pace-capture mapping along the length of the line were performed immediately, and at 2 months, post-ablation. Animals were euthanized and gap sizes were measured macroscopically. Voltage thresholds to define scar were determined by receiver operating characteristic analysis as voltage, pace-capture, and ablation contact force maps. All modalities overestimated chronic gap size, by 1.4 ± 2.0 mm (ablation contact force map), 5.1 ± 3.4 mm (pace-capture), and 9.5 ± 3.8 mm (voltage mapping). Error on ablation contact force map gap measurements were significantly less than for voltage mapping (P = 0.003, Tukey's multiple comparisons test). Chronically, voltage mapping and pace-capture mapping overestimated macroscopic gap size by 11.9 ± 3.7 and 9.8 ± 3.5 mm, respectively. Bipolar voltage and pace-capture mapping overestimate the size of chronic gap formation in linear ablation lesions. The most accurate estimation of chronic gap size was achieved by analysis of catheter-myocardium contact force during ablation.

  12. Accumulation of dsRNA in endosomes contributes to inefficient RNA interference in the fall armyworm, Spodoptera frugiperda.

    Science.gov (United States)

    Yoon, June-Sun; Gurusamy, Dhandapani; Palli, Subba Reddy

    2017-11-01

    RNA interference (RNAi) efficiency varies among insects studied. The barriers for successful RNAi include the presence of double-stranded ribonucleases (dsRNase) in the lumen and hemolymph that could potentially digest double-stranded RNA (dsRNA) and the variability in the transport of dsRNA into and within the cells. We recently showed that the dsRNAs are transported into lepidopteran cells, but they are not processed into small interference RNAs (siRNAs) because they are trapped in acidic bodies. In the current study, we focused on the identification of acidic bodies in which dsRNAs accumulate in Sf9 cells. Time-lapse imaging studies showed that dsRNAs enter Sf9 cells and accumulate in acidic bodies within 20 min after their addition to the medium. CypHer-5E-labeled dsRNA also accumulated in the midgut and fat body dissected from Spodoptera frugiperda larvae with similar patterns observed in Sf9 cells. Pharmacological inhibitor assays showed that the dsRNAs use clathrin mediated endocytosis pathway for transport into the cells. We investigated the potential dsRNA accumulation sites employing LysoTracker and double labeling experiments using the constructs to express a fusion of green fluorescence protein with early or late endosomal marker proteins and CypHer-5E-labeled dsRNA. Interestingly, CypHer-5E-labeled dsRNA accumulated predominantly in early and late endosomes. These data suggest that entrapment of internalized dsRNA in endosomes is one of the major factors contributing to inefficient RNAi response in lepidopteran insects. Copyright © 2017 Elsevier Ltd. All rights reserved.

  13. Arf6-Dependent Intracellular Trafficking of Pasteurella multocida Toxin and pH-Dependent Translocation from Late Endosomes

    Directory of Open Access Journals (Sweden)

    Tracy P. M. Chong

    2011-03-01

    Full Text Available The potent mitogenic toxin from Pasteurella multocida (PMT is the major virulence factor associated with a number of epizootic and zoonotic diseases caused by infection with this respiratory pathogen. PMT is a glutamine-specific protein deamidase that acts on its intracellular G-protein targets to increase intracellular calcium, cytoskeletal, and mitogenic signaling. PMT enters cells through receptor-mediated endocytosis and then translocates into the cytosol through a pH-dependent process that is inhibited by NH4Cl or bafilomycin A1. However, the detailed mechanisms that govern cellular entry, trafficking, and translocation of PMT remain unclear. Co-localization studies described herein revealed that while PMT shares an initial entry pathway with transferrin (Tfn and cholera toxin (CT, the trafficking pathways of Tfn, CT, and PMT subsequently diverge, as Tfn is trafficked to recycling endosomes, CT is trafficked retrograde to the ER, and PMT is trafficked to late endosomes. Our studies implicate the small regulatory GTPase Arf6 in the endocytic trafficking of PMT. Translocation of PMT from the endocytic vesicle occurs through a pH-dependent process that is also dependent on both microtubule and actin dynamics, as evidenced by inhibition of PMT activity in our SRE-based reporter assay, with nocodazole and cytochalasin D, respectively, suggesting that membrane translocation and cytotoxicity of PMT is dependent on its transfer to late endosomal compartments. In contrast, disruption of Golgi-ER trafficking with brefeldin A increased PMT activity, suggesting that inhibiting PMT trafficking to non-productive compartments that do not lead to translocation, while promoting formation of an acidic tubulovesicle system more conducive to translocation, enhances PMT translocation and activity.

  14. The late endosomal HOPS complex anchors active G-protein signaling essential for pathogenesis in magnaporthe oryzae.

    Directory of Open Access Journals (Sweden)

    Ravikrishna Ramanujam

    Full Text Available In Magnaporthe oryzae, the causal ascomycete of the devastating rice blast disease, the conidial germ tube tip must sense and respond to a wide array of requisite cues from the host in order to switch from polarized to isotropic growth, ultimately forming the dome-shaped infection cell known as the appressorium. Although the role for G-protein mediated Cyclic AMP signaling in appressorium formation was first identified almost two decades ago, little is known about the spatio-temporal dynamics of the cascade and how the signal is transmitted through the intracellular network during cell growth and morphogenesis. In this study, we demonstrate that the late endosomal compartments, comprising of a PI3P-rich (Phosphatidylinositol 3-phosphate highly dynamic tubulo-vesicular network, scaffold active MagA/GαS, Rgs1 (a GAP for MagA, Adenylate cyclase and Pth11 (a non-canonical GPCR in the likely absence of AKAP-like anchors during early pathogenic development in M. oryzae. Loss of HOPS component Vps39 and consequently the late endosomal function caused a disruption of adenylate cyclase localization, cAMP signaling and appressorium formation. Remarkably, exogenous cAMP rescued the appressorium formation defects associated with VPS39 deletion in M. oryzae. We propose that sequestration of key G-protein signaling components on dynamic late endosomes and/or endolysosomes, provides an effective molecular means to compartmentalize and control the spatio-temporal activation and rapid downregulation (likely via vacuolar degradation of cAMP signaling amidst changing cellular geometry during pathogenic development in M. oryzae.

  15. The Processed Amino-Terminal Fragment of Human TLR7 Acts as a Chaperone To Direct Human TLR7 into Endosomes

    Science.gov (United States)

    Shepherd, Dawn; Booth, Sarah; Waithe, Dominic; Reis e Sousa, Caetano

    2015-01-01

    TLR7 mediates innate immune responses to viral RNA in endocytic compartments. Mouse and human (h)TLR7 undergo proteolytic cleavage, resulting in the generation of a C-terminal fragment that accumulates in endosomes and associates with the signaling adaptor MyD88 upon receptor triggering by TLR7 agonists. Although mouse TLR7 is cleaved in endosomes by acidic proteases, hTLR7 processing can occur at neutral pH throughout the secretory pathway through the activity of furin-like proprotein convertases. However, the mechanisms by which cleaved hTLR7 reaches the endosomal compartment remain unclear. In this study, we demonstrate that, after hTLR7 proteolytic processing, the liberated amino (N)-terminal fragment remains bound to the C terminus through disulfide bonds and provides key trafficking information that ensures correct delivery of the complex to endosomal compartments. In the absence of the N-terminal fragment, the C-terminal fragment is redirected to the cell surface, where it is functionally inactive. Our data reveal a novel role for the N terminus of hTLR7 as a molecular chaperone that provides processed hTLR7 with the correct targeting instructions to reach the endosomal compartment, hence ensuring its biological activity and preventing inadvertent cell surface responses to self-RNA. PMID:25917086

  16. The PDZ protein GIPC regulates trafficking of the LPA1 receptor from APPL signaling endosomes and attenuates the cell's response to LPA.

    Directory of Open Access Journals (Sweden)

    Tal Varsano

    Full Text Available Lysophosphatidic acid (LPA mediates diverse cellular responses through the activation of at least six LPA receptors--LPA(1-6, but the interacting proteins and signaling pathways that mediate the specificity of these receptors are largely unknown. We noticed that LPA(1 contains a PDZ binding motif (SVV identical to that present in two other proteins that interact with the PDZ protein GIPC. GIPC is involved in endocytic trafficking of several receptors including TrkA, VEGFR2, lutropin and dopamine D2 receptors. Here we show that GIPC binds directly to the PDZ binding motif of LPA(1 but not that of other LPA receptors. LPA(1 colocalizes and coimmunoprecipitates with GIPC and its binding partner APPL, an activator of Akt signaling found on APPL signaling endosomes. GIPC depletion by siRNA disturbed trafficking of LPA(1 to EEA1 early endosomes and promoted LPA(1 mediated Akt signaling, cell proliferation, and cell motility. We propose that GIPC binds LPA(1 and promotes its trafficking from APPL-containing signaling endosomes to EEA1 early endosomes and thus attenuates LPA-mediated Akt signaling from APPL endosomes.

  17. The translocon protein Sec61 mediates antigen transport from endosomes in the cytosol for cross-presentation to CD8(+) T cells.

    Science.gov (United States)

    Zehner, Matthias; Marschall, Andrea L; Bos, Erik; Schloetel, Jan-Gero; Kreer, Christoph; Fehrenschild, Dagmar; Limmer, Andreas; Ossendorp, Ferry; Lang, Thorsten; Koster, Abraham J; Dübel, Stefan; Burgdorf, Sven

    2015-05-19

    The molecular mechanisms regulating antigen translocation into the cytosol for cross-presentation are under controversial debate, mainly because direct data is lacking. Here, we have provided direct evidence that the activity of the endoplasmic reticulum (ER) translocon protein Sec61 is essential for endosome-to-cytosol translocation. We generated a Sec61-specific intrabody, a crucial tool that trapped Sec61 in the ER and prevented its recruitment into endosomes without influencing Sec61 activity and antigen presentation in the ER. Expression of this ER intrabody inhibited antigen translocation and cross-presentation, demonstrating that endosomal Sec61 indeed mediates antigen transport across endosomal membranes. Moreover, we showed that the recruitment of Sec61 toward endosomes, and hence antigen translocation and cross-presentation, is dependent on dendritic cell activation by Toll-like receptor (TLR) ligands. These data shed light on a long-lasting question regarding antigen cross-presentation and point out a role of the ER-associated degradation machinery in compartments distinct from the ER. Copyright © 2015 Elsevier Inc. All rights reserved.

  18. Role of the Small GTPase Rho3 in Golgi/Endosome trafficking through functional interaction with adaptin in Fission Yeast.

    Directory of Open Access Journals (Sweden)

    Ayako Kita

    Full Text Available BACKGROUND: We had previously identified the mutant allele of apm1(+ that encodes a homolog of the mammalian µ1A subunit of the clathrin-associated adaptor protein-1 (AP-1 complex, and we demonstrated the role of Apm1 in Golgi/endosome trafficking, secretion, and vacuole fusion in fission yeast. METHODOLOGY/PRINCIPAL FINDINGS: In the present study, we isolated rho3(+, which encodes a Rho-family small GTPase, an important regulator of exocystosis, as a multicopy-suppressor of the temperature-sensitive growth of the apm1-1 mutant cells. Overexpression of Rho3 suppressed the Cl(- sensitivity and immunosuppressant sensitivity of the apm1-1 mutant cells. Overexpression of Rho3 also suppressed the fragmentation of vacuoles, and the accumulation of v-SNARE Syb1 in Golgi/endosomes and partially suppressed the defective secretion associated with apm1-deletion cells. Notably, electron microscopic observation of the rho3-deletion cells revealed the accumulation of abnormal Golgi-like structures, vacuole fragmentation, and accumulation of secretory vesicles; these phenotypes were very similar to those of the apm1-deletion cells. Furthermore, the rho3-deletion cells and apm1-deletion cells showed very similar phenotypic characteristics, including the sensitivity to the immunosuppressant FK506, the cell wall-damaging agent micafungin, Cl(-, and valproic acid. Green fluorescent protein (GFP-Rho3 was localized at Golgi/endosomes as well as the plasma membrane and division site. Finally, Rho3 was shown to form a complex with Apm1 as well as with other subunits of the clathrin-associated AP-1 complex in a GTP- and effector domain-dependent manner. CONCLUSIONS/SIGNIFICANCE: Taken together, our findings reveal a novel role of Rho3 in the regulation of Golgi/endosome trafficking and suggest that clathrin-associated adaptor protein-1 and Rho3 co-ordinate in intracellular transport in fission yeast. To the best of our knowledge, this study provides the first evidence

  19. Thyroid radiofrequency ablation: Updates on innovative devices and techniques

    Energy Technology Data Exchange (ETDEWEB)

    Park, Hye Sun; Baek, Jung Hwan; Chung, Sae Rom; Choi, Young Jun; Lee, Jeong Hyun [Dept. of Radiology and Research Institute of Radiology, University of Ulsan College of Medicine, Asan Medical Center, Seoul (Korea, Republic of); Park, Auh Whan [Dept. of Radiology, Vascular and Interventional Radiology Section, University of Virginia Health System, Charlottesville (United States)

    2017-08-01

    Radiofrequency ablation (RFA) is a well-known, effective, and safe method for treating benign thyroid nodules and recurrent thyroid cancers. Thyroid-dedicated devices and basic techniques for thyroid RFA were introduced by the Korean Society of Thyroid Radiology (KSThR) in 2012. Thyroid RFA has now been adopted worldwide, with subsequent advances in devices and techniques. To optimize the treatment efficacy and patient safety, understanding the basic and advanced RFA techniques and selecting the optimal treatment strategy are critical. The goal of this review is to therefore provide updates and analysis of current devices and advanced techniques for RFA treatment of benign thyroid nodules and recurrent thyroid cancers.

  20. Thoracoscopic sympathectomy ganglia ablation in the management ...

    African Journals Online (AJOL)

    Thoracoscopic sympathectomy ganglia ablation in the management of palmer hyperhidrosis: A decade experience in a single institution. D Kravarusic, E Freud. Abstract. Background: Hyperhidrosis can cause significant professional and social handicaps. Surgery is the preferred treatment modality for hyperhidrosis.

  1. Laser ablation in analytical chemistry - A review

    Energy Technology Data Exchange (ETDEWEB)

    Russo, Richard E.; Mao, Xianglei; Liu, Haichen; Gonzalez, Jhanis; Mao, Samuel S.

    2001-10-10

    Laser ablation is becoming a dominant technology for direct solid sampling in analytical chemistry. Laser ablation refers to the process in which an intense burst of energy delivered by a short laser pulse is used to sample (remove a portion of) a material. The advantages of laser ablation chemical analysis include direct characterization of solids, no chemical procedures for dissolution, reduced risk of contamination or sample loss, analysis of very small samples not separable for solution analysis, and determination of spatial distributions of elemental composition. This review describes recent research to understand and utilize laser ablation for direct solid sampling, with emphasis on sample introduction to an inductively coupled plasma (ICP). Current research related to contemporary experimental systems, calibration and optimization, and fractionation is discussed, with a summary of applications in several areas.

  2. Simple spherical ablative-implosion model

    International Nuclear Information System (INIS)

    Mayer, F.J.; Steele, J.T.; Larsen, J.T.

    1980-01-01

    A simple model of the ablative implosion of a high-aspect-ratio (shell radius to shell thickness ratio) spherical shell is described. The model is similar in spirit to Rosenbluth's snowplow model. The scaling of the implosion time was determined in terms of the ablation pressure and the shell parameters such as diameter, wall thickness, and shell density, and compared these to complete hydrodynamic code calculations. The energy transfer efficiency from ablation pressure to shell implosion kinetic energy was examined and found to be very efficient. It may be possible to attach a simple heat-transport calculation to our implosion model to describe the laser-driven ablation-implosion process. The model may be useful for determining other energy driven (e.g., ion beam) implosion scaling

  3. Endometrial ablation: normal appearance and complications.

    Science.gov (United States)

    Drylewicz, Monica R; Robinson, Kathryn; Siegel, Cary Lynn

    2018-03-14

    Global endometrial ablation is a commonly performed, minimally invasive technique aimed at improving/resolving abnormal uterine bleeding and menorrhagia in women. As non-resectoscopic techniques have come into existence, endometrial ablation performance continues to increase due to accessibility and decreased requirements for operating room time and advanced technical training. The increased utilization of this method translates into increased imaging of patients who have undergone the procedure. An understanding of the expected imaging appearances of endometrial ablation using different modalities is important for the abdominal radiologist. In addition, the frequent usage of the technique naturally comes with complications requiring appropriate imaging work-up. We review the expected appearance of the post-endometrial ablated uterus on multiple imaging modalities and demonstrate the more common and rare complications seen in the immediate post-procedural time period and remotely.

  4. Microwave Tissue Ablation: Biophysics, Technology and Applications

    Science.gov (United States)

    2010-01-01

    Microwave ablation is an emerging treatment option for many cancers, cardiac arrhythmias and other medical conditions. During treatment, microwaves are applied directly to tissues to produce rapid temperature elevations sufficient to produce immediate coagulative necrosis. The engineering design criteria for each application differ, with individual consideration for factors such as desired ablation zone size, treatment duration, and procedural invasiveness. Recent technological developments in applicator cooling, power control and system optimization for specific applications promise to increase the utilization of microwave ablation in the future. This article will review the basic biophysics of microwave tissue heating, provide an overview of the design and operation of current equipment, and outline areas for future research for microwave ablation. PMID:21175404

  5. Diagnostics of laser ablated plasma plumes

    DEFF Research Database (Denmark)

    Amoruso, S.; Toftmann, B.; Schou, Jørgen

    2004-01-01

    The effect of an ambient gas on the expansion dynamics of laser ablated plasmas has been studied for two systems by exploiting different diagnostic techniques. First, the dynamics of a MgB2 laser produced plasma plume in an Ar atmosphere has been investigated by space-and time-resolved optical...... of the laser ablated plasma plume propagation in a background gas. (C) 2003 Elsevier B.V All rights reserved....

  6. ROLE OF RADIOFREQUENCY ABLATION IN ADENOMA SEBACEUM

    Directory of Open Access Journals (Sweden)

    Ch. Madh

    2016-03-01

    Full Text Available Adenoma sebaceum, pathognomonic of tuberous sclerosis, are tiny angiofibromas which commonly occur over central part of face. Recurrence after treatment is common and hence a need for inexpensive, safe and efficient treatment is required. Radiofrequency ablation is a safe and an economical procedure and has been known to cause less scarring with good aesthetic results compared to other ablative methods such as electrocautery.

  7. Micrometeoroid ablation simulated in the laboratory

    Science.gov (United States)

    Sternovsky, Zoltan; Thomas, Evan W.; DeLuca, Michael; Horanyi, Mihaly; Janches, Diego; Munsat, Tobin L.; Plane, John M. C.

    2016-04-01

    A facility is developed to simulate the ablation of micrometeoroids in laboratory conditions, which also allows measuring the ionization probability of the ablated material. An electrostatic dust accelerator is used to generate iron and meteoric analog particles with velocities 10-50 km/s. The particles are then introduced into a cell filled with nitrogen, air or carbon dioxide gas with pressures adjustable in the 0.02 - 0.5 Torr range, where the partial or complete ablation of the particle occurs over a short distance. An array of biased electrodes is used to collect the ionized products with spatial resolution along the ablating particles' path, allowing thus the study of the temporal resolution of the process. A simple ablation model is used to match the observations. For completely ablated particles the total collected charge directly yields the ionization efficiency for. The measurements using iron particles in N2 and air are in relatively good agreement with earlier data. The measurements with CO2 and He gases, however, are significantly different from the expectations.

  8. Laser ablation of microparticles for nanostructure generation

    International Nuclear Information System (INIS)

    Waraich, Palneet Singh; Tan, Bo; Venkatakrishnan, Krishnan

    2011-01-01

    The process of laser ablation of microparticles has been shown to generate nanoparticles from microparticles; but the generation of nanoparticle networks from microparticles has never been reported before. We report a unique approach for the generation of nanoparticle networks through ablation of microparticles. Using this approach, two samples containing microparticles of lead oxide (Pb 3 O 4 ) and nickel oxide (NiO), respectively, were ablated under ambient conditions using a femtosecond laser operating in the MHz repetition rate regime. Nanoparticle networks with particle diameter ranging from 60 to 90 nm were obtained by ablation of microparticles without use of any specialized equipment, catalysts or external stimulants. The formation of finer nanoparticle networks has been explained by considering the low pressure region created by the shockwave, causing rapid condensation of microparticles into finer nanoparticles. A comparison between the nanostructures generated by ablating microparticle and those by ablating bulk substrate was carried out; and a considerable reduction in size and narrowed size distribution was observed. Our nanostructure fabrication technique will be a unique process for nanoparticle network generation from a vast array of materials.

  9. Combinations of ERK and p38 MAPK inhibitors ablate tumor necrosis factor-alpha (TNF-alpha ) mRNA induction. Evidence for selective destabilization of TNF-alpha transcripts.

    Science.gov (United States)

    Rutault, K; Hazzalin, C A; Mahadevan, L C

    2001-03-02

    Tumor necrosis factor-alpha (TNF-alpha) is a potent proinflammatory cytokine whose synthesis and secretion are implicated in diverse pathologies. Hence, inhibition of TNF-alpha transcription or translation and neutralization of its protein product represent major pharmaceutical strategies to control inflammation. We have studied the role of ERK and p38 mitogen-activated protein (MAP) kinase in controlling TNF-alpha mRNA levels in differentiated THP-1 cells and in freshly purified human monocytes. We show here that it is possible to produce virtually complete inhibition of lipopolysaccharide-stimulated TNF-alpha mRNA accumulation by using a combination of ERK and p38 MAP kinase inhibitors. Furthermore, substantial inhibition is achievable using combinations of 1 microm of each inhibitor, whereas inhibitors used individually are incapable of producing complete inhibition even at high concentrations. Finally, addressing mechanisms involved, we show that inhibition of p38 MAP kinase selectively destabilizes TNF-alpha transcripts but does not affect degradation of c-jun transcripts. These results impinge on the controversy in the literature surrounding the mode of action of MAP kinase inhibitors on TNF-alpha mRNA and suggest the use of combinations of MAP kinase inhibitors as an effective anti-inflammatory strategy.

  10. Reorganization of the Endosomal System in Salmonella-Infected Cells: The Ultrastructure of Salmonella-Induced Tubular Compartments

    Science.gov (United States)

    Krieger, Viktoria; Liebl, David; Zhang, Yuying; Rajashekar, Roopa; Chlanda, Petr; Giesker, Katrin; Chikkaballi, Deepak; Hensel, Michael

    2014-01-01

    During the intracellular life of Salmonella enterica, a unique membrane-bound compartment termed Salmonella-containing vacuole, or SCV, is formed. By means of translocated effector proteins, intracellular Salmonella also induce the formation of extensive, highly dynamic membrane tubules termed Salmonella-induced filaments or SIF. Here we report the first detailed ultrastructural analyses of the SCV and SIF by electron microscopy (EM), EM tomography and live cell correlative light and electron microscopy (CLEM). We found that a subset of SIF is composed of double membranes that enclose portions of host cell cytosol and cytoskeletal filaments within its inner lumen. Despite some morphological similarities, we found that the formation of SIF double membranes is independent from autophagy and requires the function of the effector proteins SseF and SseG. The lumen of SIF network is accessible to various types of endocytosed material and our CLEM analysis of double membrane SIF demonstrated that fluid phase markers accumulate only between the inner and outer membrane of these structures, a space continual with endosomal lumen. Our work reveals how manipulation of the endosomal membrane system by an intracellular pathogen results in a unique tubular membrane compartmentalization of the host cell, generating a shielded niche permissive for intracellular proliferation of Salmonella. PMID:25254663

  11. Multi-layered nanoparticles for penetrating the endosome and nuclear membrane via a step-wise membrane fusion process.

    Science.gov (United States)

    Akita, Hidetaka; Kudo, Asako; Minoura, Arisa; Yamaguti, Masaya; Khalil, Ikramy A; Moriguchi, Rumiko; Masuda, Tomoya; Danev, Radostin; Nagayama, Kuniaki; Kogure, Kentaro; Harashima, Hideyoshi

    2009-05-01

    Efficient targeting of DNA to the nucleus is a prerequisite for effective gene therapy. The gene-delivery vehicle must penetrate through the plasma membrane, and the DNA-impermeable double-membraned nuclear envelope, and deposit its DNA cargo in a form ready for transcription. Here we introduce a concept for overcoming intracellular membrane barriers that involves step-wise membrane fusion. To achieve this, a nanotechnology was developed that creates a multi-layered nanoparticle, which we refer to as a Tetra-lamellar Multi-functional Envelope-type Nano Device (T-MEND). The critical structural elements of the T-MEND are a DNA-polycation condensed core coated with two nuclear membrane-fusogenic inner envelopes and two endosome-fusogenic outer envelopes, which are shed in stepwise fashion. A double-lamellar membrane structure is required for nuclear delivery via the stepwise fusion of double layered nuclear membrane structure. Intracellular membrane fusions to endosomes and nuclear membranes were verified by spectral imaging of fluorescence resonance energy transfer (FRET) between donor and acceptor fluorophores that had been dually labeled on the liposome surface. Coating the core with the minimum number of nucleus-fusogenic lipid envelopes (i.e., 2) is essential to facilitate transcription. As a result, the T-MEND achieves dramatic levels of transgene expression in non-dividing cells.

  12. Percutaneous Microwave Ablation of Renal Angiomyolipomas

    Energy Technology Data Exchange (ETDEWEB)

    Cristescu, Mircea, E-mail: mcristescu@uwhealth.org [University of Wisconsin, Department of Radiology (United States); Abel, E. Jason, E-mail: abel@urology.wisc.edu [University of Wisconsin, Department of Urology (United States); Wells, Shane, E-mail: swells@uwhealth.org; Ziemlewicz, Timothy J., E-mail: tziemlewicz@uwhealth.org [University of Wisconsin, Department of Radiology (United States); Hedican, Sean P., E-mail: hedican@surgery.wisc.edu [University of Wisconsin, Department of Urology (United States); Lubner, Megan G., E-mail: mlubner@uwhealth.org; Hinshaw, J. Louis, E-mail: jhinshaw@uwhealth.org; Brace, Christopher L., E-mail: cbrace@uwhealth.org; Lee, Fred T., E-mail: flee@uwhealth.org [University of Wisconsin, Department of Radiology (United States)

    2016-03-15

    PurposeTo evaluate the safety and efficacy of US-guided percutaneous microwave (MW) ablation in the treatment of renal angiomyolipoma (AML).Materials and MethodsFrom January 2011 to April 2014, seven patients (5 females and 2 males; mean age 51.4) with 11 renal AMLs (9 sporadic type and 2 tuberous sclerosis associated) with a mean size of 3.4 ± 0.7 cm (range 2.4–4.9 cm) were treated with high-powered, gas-cooled percutaneous MW ablation under US guidance. Tumoral diameter, volume, and CT/MR enhancement were measured on pre-treatment, immediate post-ablation, and delayed post-ablation imaging. Clinical symptoms and creatinine were assessed on follow-up visits.ResultsAll ablations were technically successful and no major complications were encountered. Mean ablation parameters were ablation power of 65 W (range 60–70 W), using 456 mL of hydrodissection fluid per patient, over 4.7 min (range 3–8 min). Immediate post-ablation imaging demonstrated mean tumor diameter and volume decreases of 1.8 % (3.4–3.3 cm) and 1.7 % (27.5–26.3 cm{sup 3}), respectively. Delayed imaging follow-up obtained at a mean interval of 23.1 months (median 17.6; range 9–47) demonstrated mean tumor diameter and volume decreases of 29 % (3.4–2.4 cm) and 47 % (27.5–12.1 cm{sup 3}), respectively. Tumoral enhancement decreased on immediate post-procedure and delayed imaging by CT/MR parameters, indicating decreased tumor vascularity. No patients required additional intervention and no patients experienced spontaneous bleeding post-ablation.ConclusionOur early experience with high-powered, gas-cooled percutaneous MW ablation demonstrates it to be a safe and effective modality to devascularize and decrease the size of renal AMLs.

  13. Effect of ablatant composition on the ablation of a fuelling pellet

    International Nuclear Information System (INIS)

    Chang, C.T.; Thomsen, K.; Piret, S.

    1988-01-01

    The single species neutral-shielding model for the ablation of a hydrogenic pellet is extended by considering the ablatant as a mixture of four species: molecular and atomic hydrogen, protons and electrons. Compared with the results of the frozen flow, (i.e. the single species molecular hydrogen gas model), results of the analysis showed that the presence of dissociation and ionization effects caused a marked difference of the ablatant state. The attenuations of the incoming electron energy and energy flux, however, are very much similar irrespective of whether the ablated flow is in a frozen or an equilibrium state. The scaling law of the pellet ablation rate with respect to the plasma state of Te, ne and the pellet radius remains the same; the ablation rate is reduced by approximately 15%. To examine the possible existence of a spherical shell around the pellet where most of the incoming electron energy is absorbed, acodmparison is made between the local electron collisional mean free path and the electron Larmor radius. A critical field at the ionization radius is evaluated. An effective spherical energyabsorbing region exists when the local field strength is below the critical value. For a plasma state of low Te and ne, (where the ablatant is hardly ionized), and for one near the thermonuclear condition (where a highly dense ablatant exists near the pellet), the effective energy absorption region is nearly spherical. 20 refs. (author)

  14. Ablation mass features in multi-pulses femtosecond laser ablate molybdenum target

    Science.gov (United States)

    Zhao, Dongye; Gierse, Niels; Wegner, Julian; Pretzler, Georg; Oelmann, Jannis; Brezinsek, Sebastijan; Liang, Yunfeng; Neubauer, Olaf; Rasinski, Marcin; Linsmeier, Christian; Ding, Hongbin

    2018-03-01

    In this study, the ablation mass features related to reflectivity of bulk Molybdenum (Mo) were investigated by a Ti: Sa 6 fs laser pulse at central wavelength 790 nm. The ablated mass removal was determined using Confocal Microscopy (CM) technique. The surface reflectivity was calibrated and measured by a Lambda 950 spectrophotometer as well as a CCD camera during laser ablation. The ablation mass loss per pulse increase with the increasing of laser shots, meanwhile the surface reflectivity decrease. The multi-pulses (100 shots) ablation threshold of Mo was determined to be 0.15 J/cm2. The incubation coefficient was estimated as 0.835. The reflectivity change of the Mo target surface following multi-pulses laser ablation were studied as a function of laser ablation shots at various laser fluences from 1.07 J/cm2 to 36.23 J/cm2. The results of measured reflectivity indicate that surface reflectivity of Mo target has a significant decline in the first 3-laser pulses at the various fluences. These results are important for developing a quantitative analysis model for laser induced ablation and laser induced breakdown spectroscopy for the first wall diagnosis of EAST tokamak.

  15. Fractional ablative erbium YAG laser

    DEFF Research Database (Denmark)

    Taudorf, Elisabeth H; Haak, Christina S; Erlendsson, Andrés M

    2014-01-01

    laser parameters with tissue effects. MATERIALS AND METHODS: Ex vivo pig skin was exposed to a miniaturized 2,940 nm AFXL, spot size 225 µm, density 5%, power levels 1.15-2.22 W, pulse durations 50-225 microseconds, pulse repetition rates 100-500 Hz, and 2, 20, or 50 stacked pulses, resulting in pulse......BACKGROUND AND OBJECTIVES: Treatment of a variety of skin disorders with ablative fractional lasers (AFXL) is driving the development of portable AFXLs. This study measures micropore dimensions produced by a small 2,940 nm AFXL using a variety of stacked pulses, and determines a model correlating...... 190 to 347 µm. CONCLUSIONS: Pulse stacking with a small, low power 2,940 nm AFXL created reproducible shallow to deep micropores, and influenced micropore configuration. Mathematical modeling established relations between laser settings and micropore dimensions, which assists in choosing laser...

  16. Radiofrequency thermal ablation of malignant hepatic tumors: post-ablation syndrome

    International Nuclear Information System (INIS)

    Choi, Jung Bin; Rhim, Hyunchul; Kim, Yongsoo; Koh, Byung Hee; Cho, On Koo; Seo, Heung Suk; Lee, Seung Ro

    2000-01-01

    To evaluate post-ablation syndrome after radiofrequency thermal ablation of malignant hepatic tumors. Forty-two patients with primary (n=3D29) or secondary (n=3D13) hepatic tumors underwent radiofrequency thermal ablation. A total of 65 nodules ranging in size from 1.1 to 5.0 (mean, 3.1) cm were treated percutaneously using a 50W RF generator with 15G expandable needle electrodes. We retrospectively evaluated the spectrum of post-ablation syndrome including pain, fever (≥3D 38 deg C), nausea, vomiting, right shoulder pain, and chest discomfort according to frequency, intensity and duration, and the findings were correlated with tumor location and number of ablations. We also evaluated changes in pre-/post-ablation serum aminotransferase (ALT/AST) and prothrombin time, and correlated these findings with the number of ablations. Post-ablation syndrome was noted in 29 of 42 patients (69.0%), and most symptoms improved with conservative treatment. The most important of these were abdominal plan (n=3D20, 47.6%), fever (n=3D8, 19.0%), and nausea (n=3D7, 16.7%), and four of 42 (9.5%) patients complained of severe pain. The abdominal pain lasted from 3 hours to 5.5 days (mean; 20.4 hours), the fever from 6 hours to 5 days (mean; 63.0 hours). And the nausea from 1 hours to 4 days (mean; 21.0 hours). Other symptoms were right shoulder pain (n=3D6, 14.3%), chest discomfort (n=3D3, 7.1%), and headache (n=3D3, 7.1%). Seventeen of 20 patients (85%) with abdominal pain had subcapsular tumor of the liver. There was significant correlation between pain, location of the tumor, and a number of ablations. After ablation, ALT/AST was elevated more than two-fold in 52.6%/73.7% of patients, respectively but there was no significant correlation with the number of ablation. Post-ablation syndrome is a frequent and tolerable post-procedural process after radiofrequency thermal ablation. The spectrum of this syndrome provides a useful guideline for the post-ablation management. (author)

  17. Endovenous Laser Ablation as a Treatment for Postsurgical Recurrent Saphenous Insufficiency

    International Nuclear Information System (INIS)

    Anchala, Praveen R.; Wickman, Christopher; Chen, Richard; Faundeen, Tonya; Pearce, William; Narducy, Lisa; Resnick, Scott A.

    2010-01-01

    The purpose of this study was to investigate the safety and efficacy of endovenous laser ablation as a treatment for recurrent symptomatic saphenous insufficiency occurring after saphenous vein ligation and stripping. A single-center retrospective review of patients who received endovenous laser ablation as a treatment for recurrent symptomatic saphenous insufficiency after ligation and stripping between November 2003 and October 2006 was performed. Fifty-six insufficient saphenous systems were identified in 38 patients. Follow-up consisted of a clinical examination in all patients as well as selective lower-extremity duplex ultrasound as clinically indicated. All 38 patients demonstrated complete closure of the insufficient saphenous vein by clinical examination and/or duplex ultrasound evaluation. Preoperative symptoms resolved after treatment in all 38 patients. No major complications were identified. Endovenous laser ablation of recurrent symptomatic saphenous venous insufficiency is a safe and effective treatment in patients who develop recurrent symptoms after saphenous vein ligation and stripping.

  18. Synthesis of higher diamondoids by pulsed laser ablation plasmas in supercritical CO2

    International Nuclear Information System (INIS)

    Nakahara, Sho; Stauss, Sven; Kato, Toru; Terashima, Kazuo; Sasaki, Takehiko

    2011-01-01

    Pulsed laser ablation (wavelength 532 nm; fluence 18 J/cm 2 ; pulse width 7 ns; repetition rate 10 Hz) of highly oriented pyrolytic graphite was conducted in adamantane-dissolved supercritical CO 2 with and without cyclohexane as a cosolvent. Micro-Raman spectroscopy of the products revealed the presence of hydrocarbons possessing sp 3 -hybridized carbons similar to diamond structures. The synthesis of diamantane and other possible diamondoids consisting of up to 12 cages was confirmed by gas chromatography-mass spectrometry. Furthermore, gas chromatography-mass spectrometry measurements of samples before and after pyrolysis treatment indicate the synthesis of the most compact decamantane, namely, superadamantane. It is thought that oxidant species originating from CO 2 during pulsed laser ablation might lead to the selective dissociation of C-H bonds, enabling the synthesis of low H/C ratio molecules. Therefore, laser ablation in supercritical CO 2 is proposed as a practical method for synthesizing diamondoids.

  19. CuInGaSe{sub 2} nanoparticles by pulsed laser ablation in liquid medium

    Energy Technology Data Exchange (ETDEWEB)

    Mendivil, M.I.; García, L.V. [Facultad de Ingeniería Mecánica y Eléctrica, Universidad Autónoma de Nuevo León, San Nicolás de los Garza, Nuevo León, 66455 (Mexico); Krishnan, B. [Facultad de Ingeniería Mecánica y Eléctrica, Universidad Autónoma de Nuevo León, San Nicolás de los Garza, Nuevo León, 66455 (Mexico); CIIDIT—Universidad Autónoma de Nuevo León, Apodaca, Nuevo León (Mexico); Avellaneda, D. [Facultad de Ingeniería Mecánica y Eléctrica, Universidad Autónoma de Nuevo León, San Nicolás de los Garza, Nuevo León, 66455 (Mexico); and others

    2015-12-15

    Highlights: • CIGS nanocolloids were synthesized using PLAL technique. • Characterized their morphology, structure, composition and optical properties. • Morphologies were dependent on ablation wavelength and liquid medium. • Optical absorption and bandgap of these nanocolloids were tunable. - Abstract: Pulsed laser ablation in liquid medium (PLALM) is a nanofabrication technique to produce complex nanostructures. CuInGaSe{sub 2} (CIGS) is an alloy with applications in photovoltaic industry. In this work, we studied the effects of laser ablation wavelength, energy fluence and liquid medium on the properties of the CIGS nanoparticles synthesized by PLALM. The nanoparticles obtained were analyzed by transmission electron microscopy (TEM), energy dispersive X-ray spectroscopy (EDX), selected area electron diffraction (SAED), X-ray photoelectron spectroscopy (XPS) and UV–vis absorption spectroscopy. XPS results confirmed the chemical states and composition of the ablated products. TEM analysis showed different morphologies for the nanomaterials obtained in different liquid media and ablation wavelengths. The optical properties for these CIGS nanocolloids were analyzed using UV–vis absorption spectroscopy. The results demonstrated the use of PLALM as a useful synthesis technique for nanoparticles of quaternary photovoltaic materials.

  20. "Rescue" ablation of electrical storm in arrhythmogenic right ventricular cardiomyopathy in pregnancy.

    Science.gov (United States)

    Stec, Sebastian; Krynski, Tomasz; Baran, Jakub; Kulakowski, Piotr

    2013-08-13

    Radiofrequency ablation (RFCA) became a treatment of choice in patients with recurrent ventricular tachycardia, ventricular fibrillation, and appropriate interventions of implanted cardioverter-defibrillator (ICD), however, electrical storm (ES) ablation in a pregnant woman has not yet been reported. We describe a case of a successful rescue ablation of recurrent ES in a 26-year-old Caucasian woman during her first pregnancy (23rd week). The arrhythmogenic right ventricular dysplasia/cardiomyopathy (ARVD/C) was diagnosed 3 years earlier and several drugs as well as 2 ablations failed to control recurrences of ventricular tachycardia. RFCA was performed on the day of the third electric storm. The use of electroanatomic mapping allowed very low X-ray exposure, and after applications in the right ventricular outflow tract, arrhythmia disappeared. Three months after ablation, a healthy girl was delivered without any complications. During twelve-month follow-up there was no recurrence of ventricular tachycardia or ICD interventions. This case documents the first successful RFCA during ES due to recurrent unstable ventricular arrhythmias in a patient with ARVD/C in pregnancy. Current guidelines recommend metoprolol, sotalol and intravenous amiodarone for prevention of recurrent ventricular tachycardia in pregnancy, however, RFCA should be considered as a therapeutic option in selected cases. The use of 3D navigating system and near zero X-ray approach is associated with minimal radiation exposure for mother and fetus as well as low risk of procedural complication.

  1. Factors Limiting Complete Tumor Ablation by Radiofrequency Ablation

    International Nuclear Information System (INIS)

    Paulet, Erwan; Aube, Christophe; Pessaux, Patrick; Lebigot, Jerome; Lhermitte, Emilie; Oberti, Frederic; Ponthieux, Anne; Cales, Paul; Ridereau-Zins, Catherine; Pereira, Philippe L.

    2008-01-01

    The purpose of this study was to determine radiological or physical factors to predict the risk of residual mass or local recurrence of primary and secondary hepatic tumors treated by radiofrequency ablation (RFA). Eighty-two patients, with 146 lesions (80 hepatocellular carcinomas, 66 metastases), were treated by RFA. Morphological parameters of the lesions included size, location, number, ultrasound echogenicity, computed tomography density, and magnetic resonance signal intensity were obtained before and after treatment. Parameters of the generator were recorded during radiofrequency application. The recurrence-free group was statistically compared to the recurrence and residual mass groups on all these parameters. Twenty residual masses were detected. Twenty-nine lesions recurred after a mean follow-up of 18 months. Size was a predictive parameter. Patients' sex and age and the echogenicity and density of lesions were significantly different for the recurrence and residual mass groups compared to the recurrence-free group (p < 0.05). The presence of an enhanced ring on the magnetic resonance control was more frequent in the recurrence and residual mass groups. In the group of patients with residual lesions, analysis of physical parameters showed a significant increase (p < 0.05) in the time necessary for the temperature to rise. In conclusion, this study confirms risk factors of recurrence such as the size of the tumor and emphasizes other factors such as a posttreatment enhanced ring and an increase in the time necessary for the rise in temperature. These factors should be taken into consideration when performing RFA and during follow-up

  2. Radioiodine Remnant Ablation: A Critical Review

    International Nuclear Information System (INIS)

    Bal, Chandra Sekhar; Padhy, Ajit Kumar

    2015-01-01

    Radioiodine remnant ablation (RRA) is considered a safe and effective method for eliminating residual thyroid tissue, as well as microscopic disease if at all present in thyroid bed following thyroidectomy. The rationale of RRA is that in the absence of thyroid tissue, serum thyroglobulin (Tg) measurement can be used as an excellent tumor marker. Other considerations are like the presence of significant remnant thyroid tissue makes detection and treatment of nodal or distant metastases difficult. Rarely, microscopic disease in the thyroid bed if not ablated, in the future, could be a source of anaplastic transformation. On the other hand, microscopic tumor emboli in distant sites could be the cause of distant metastasis too. The ablation of remnant tissue would in all probability eliminate these theoretical risks. It may be noted that all these are unproven contentious issues except postablation serum Tg estimation that could be a good tumor marker for detecting early biochemical recurrence in long-term follow-up strategy. Radioactive iodine is administered as a form of “adjuvant therapy” for remnant ablation. There have been several reports with regard to the administered dose for remnant ablation. The first report of a prospective randomized clinical trial was published from India by a prospective randomized study conducted at the All India Institute of Medical Sciences, New Delhi in the year 1996. The study reported that increasing the empirical 131 I initial dose to more than 50 mCi results in plateauing of the dose-response curve and thus, conventional high-dose remnant ablation needs critical evaluation. Recently, two important studies were published: One from French group and the other from UK on a similar line. Interestingly, all three studies conducted in three different geographical regions of the world showed exactly similar conclusion. The new era of low-dose remnant ablation has taken a firm scientific footing across the continents

  3. Ablation threshold and ablation mechanism transition of polyoxymethylene irradiated by CO2 laser.

    Science.gov (United States)

    Li, Gan; Cheng, Mousen; Li, Xiaokang

    2016-09-01

    Polyoxymethylene (POM) decomposes gradually as it is heated up by the irradiation of CO2 laser; the long-chain molecules of POM are broken into short chains, which leads to the lowering of the melting point and the critical temperature of the ablation products. When the product temperature is above the melting point, ablation comes up in the way of vaporization; when the product temperature is higher than the critical temperature, all liquid products are transformed into gas instantly and the ablation mechanism is changed. The laser fluence at which significant ablation is observed is defined as the ablation threshold, and the fluence corresponding to the ablation mechanism changing is denoted as the flyover threshold. In this paper, random pyrolysis is adopted to describe the pyrolytic decomposition of POM, and consequently, the components of the pyrolysis products under different pyrolysis rates are acquired. The Group Contribution method is used to count the thermodynamic properties of the pyrolysis products, and the melting point and the critical temperature of the product mixture are obtained by the Mixing Law. The Knudsen layer relationship is employed to evaluate the ablation mass removal when the product temperature is below the critical temperature. The gas dynamics conservation laws associated with the Jouguet condition are used to calculate the mass removal when the product temperature is higher than the critical temperature. Based on the model, a set of simulations for various laser intensities and lengths are carried out to generalize the relationships between the thresholds and the laser parameters. Besides the ablated mass areal density, which fits the experimental data quite well, the ablation temperature, pyrolysis rate, and product components are also discussed for a better understanding of the ablation mechanism of POM.

  4. Assessment of laser ablation techniques in a-si technologies for position-sensor development

    Science.gov (United States)

    Molpeceres, C.; Lauzurica, S.; Ocana, J. L.; Gandia, J. J.; Urbina, L.; Carabe, J.

    2005-07-01

    Laser micromachining of semiconductor and Transparent Conductive Oxides (TCO) materials is very important for the practical applications in photovoltaic industry. In particular, a problem of controlled ablation of those materials with minimum of debris and small heat affected zone is one of the most vital for the successful implementation of laser micromachining. In particular, selective ablation of thin films for the development of new photovoltaic panels and sensoring devices based on amorphous silicon (a-Si) is an emerging field, in which laser micromachining systems appear as appropriate tools for process development and device fabrication. In particular, a promising application is the development of purely photovoltaic position sensors. Standard p-i-n or Schottky configurations using Transparent Conductive Oxides (TCO), a-Si and metals are especially well suited for these applications, appearing selective laser ablation as an ideal process for controlled material patterning and isolation. In this work a detailed study of laser ablation of a widely used TCO, Indium-tin-oxide (ITO), and a-Si thin films of different thicknesses is presented, with special emphasis on the morphological analysis of the generated grooves. The profiles of ablated grooves have been studied in order to determine the best processing conditions, i.e. laser pulse energy and wavelength, and to asses this technology as potentially competitive to standard photolithographic processes. The encouraging results obtained, with well defined ablation grooves having thicknesses in the order of 10 μm both in ITO and a-Si, open up the possibility of developing a high-performance double Schottky photovoltaic matrix position sensor.

  5. Efficacy of ablation at the anteroseptal line for the treatment of perimitral flutter

    Directory of Open Access Journals (Sweden)

    Bernard Abi-Saleh, MD, FACP, FACC, FHRS

    2015-12-01

    Conclusion: Ablation at the left atrial anteroseptal line is safe and efficacious for the treatment of PMF. Unlike ablation at the traditional mitral isthmus line, ablation at the left atrial anteroseptal line does not require ablation in the coronary sinus.

  6. Increased preference of surface ablation over laser in situ keratomileusis between 2008–2011 is correlated to risk of ecatasia

    Directory of Open Access Journals (Sweden)

    Moisseiev E

    2013-01-01

    Full Text Available Elad Moisseiev,1,3 Tzahi Sela,2 Liza Minkev,2 David Varssano1,31Department of Ophthalmology, Tel-Aviv Medical Center, Tel-Aviv, Israel; 2Care Vision, Tel Aviv, Israel; 3Sackler Faculty of Medicine, Tel Aviv University, Tel Aviv, IsraelPurpose: To evaluate the trends in corneal refractive procedure selection for the correction of myopia, focusing on the relative proportions of laser in situ keratomileusis (LASIK and surface ablation procedures.Methods: Only eyes that underwent LASIK or surface ablation for the correction of myopia between 2008–2011 were included in this retrospective study. Additional recorded parameters included patient age, preoperative manifest refraction, corneal thickness, and calculated residual corneal bed thickness. A risk score was given to each eye, based on these parameters, according to the Ectasia Risk Factor Score System (ERFSS, without the preoperative corneal topography.Results: This study included 16,163 eyes, of which 38.4% underwent LASIK and 61.6% underwent surface ablation. The risk score correlated with procedure selection, with LASIK being preferred in eyes with a score of 0 and surface ablation in eyes with a score of 2 or higher. When controlling for age, preoperative manifest refraction, corneal thickness, and all parameters, the relative proportion of surface ablation compared with LASIK was found to have grown significantly during the study period.Conclusions: Our results indicate that with time, surface ablation tended to be performed more often than LASIK for the correction of myopia in our cohort. Increased awareness of risk factors and preoperative risk assessment tools, such as the ERFSS, have shifted the current practice of refractive surgery from LASIK towards surface ablation despite the former's advantages, especially in cases in which the risk for ectasia is more than minimal (risk score 2 and higher.Keywords: surface ablation, LASIK, PRK, myopia correction, ectasia

  7. In vitro parameter optimization for spatial control of focused ultrasound ablation when using low boiling point phase-change nanoemulsions.

    Science.gov (United States)

    Puett, Connor; Phillips, Linsey C; Sheeran, Paul S; Dayton, Paul A

    2013-01-01

    ). Although the ablation lesions were located within much larger microbubble clouds, optimum insonation times and intensities could be selected to achieve an ablation lesion of desired size and location for a given PSNE concentration. This demonstration of controllable enhancement using a PSNE that contained a volatile PFC component is another step toward developing phase-shift nanotechnology as a potential clinical tool to improve HIFU.

  8. Imaging in percutaneous ablation for atrial fibrillation

    Energy Technology Data Exchange (ETDEWEB)

    Maksimovic, Ruzica [Erasmus Medical Center, Department of Radiology, GD Rotterdam (Netherlands); Institute for Cardiovascular Diseases of the University Medical Center, Belgrade (Czechoslovakia); Dill, Thorsten [Kerckhoff-Heart Center, Department of Cardiology, Bad Nauheim (Germany); Ristic, Arsen D.; Seferovic, Petar M. [Institute for Cardiovascular Diseases of the University Medical Center, Belgrade (Czechoslovakia)

    2006-11-15

    Percutaneous ablation for electrical disconnection of the arrhythmogenic foci using various forms of energy has become a well-established technique for treating atrial fibrillation (AF). Success rate in preventing recurrence of AF episodes is high although associated with a significant incidence of pulmonary vein (PV) stenosis and other rare complications. Clinical workup of AF patients includes imaging before and after ablative treatment using different noninvasive and invasive techniques such as conventional angiography, transoesophageal and intracardiac echocardiography, computed tomography (CT) and magnetic resonance imaging (MRI), which offer different information with variable diagnostic accuracy. Evaluation before percutaneous ablation involves assessment of PVs (PV pattern, branching pattern, orientation and ostial size) to facilitate position and size of catheters and reduce procedure time as well as examining the left atrium (presence of thrombi, dimensions and volumes). Imaging after the percutaneous ablation is important for assessment of overall success of the procedure and revealing potential complications. Therefore, imaging methods enable depiction of PVs and the anatomy of surrounding structures essential for preprocedural management and early detection of PV stenosis and other ablation-related procedures, as well as long-term follow-up of these patients. (orig.)

  9. Imaging in percutaneous ablation for atrial fibrillation

    International Nuclear Information System (INIS)

    Maksimovic, Ruzica; Dill, Thorsten; Ristic, Arsen D.; Seferovic, Petar M.

    2006-01-01

    Percutaneous ablation for electrical disconnection of the arrhythmogenic foci using various forms of energy has become a well-established technique for treating atrial fibrillation (AF). Success rate in preventing recurrence of AF episodes is high although associated with a significant incidence of pulmonary vein (PV) stenosis and other rare complications. Clinical workup of AF patients includes imaging before and after ablative treatment using different noninvasive and invasive techniques such as conventional angiography, transoesophageal and intracardiac echocardiography, computed tomography (CT) and magnetic resonance imaging (MRI), which offer different information with variable diagnostic accuracy. Evaluation before percutaneous ablation involves assessment of PVs (PV pattern, branching pattern, orientation and ostial size) to facilitate position and size of catheters and reduce procedure time as well as examining the left atrium (presence of thrombi, dimensions and volumes). Imaging after the percutaneous ablation is important for assessment of overall success of the procedure and revealing potential complications. Therefore, imaging methods enable depiction of PVs and the anatomy of surrounding structures essential for preprocedural management and early detection of PV stenosis and other ablation-related procedures, as well as long-term follow-up of these patients. (orig.)

  10. Fracture in Phenolic Impregnated Carbon Ablator

    Science.gov (United States)

    Agrawal, Parul; Chavez-Garcia, Jose; Pham, John

    2013-01-01

    This paper describes the development of a novel technique to understand the failure mechanisms inside thermal protection materials. The focus of this research is on the class of materials known as phenolic impregnated carbon ablators. It has successfully flown on the Stardust spacecraft and is the thermal protection system material chosen for the Mars Science Laboratory and SpaceX Dragon spacecraft. Although it has good thermal properties, structurally, it is a weak material. To understand failure mechanisms in carbon ablators, fracture tests were performed on FiberForm(Registered TradeMark) (precursor), virgin, and charred ablator materials. Several samples of these materials were tested to investigate failure mechanisms at a microstructural scale. Stress-strain data were obtained simultaneously to estimate the tensile strength and toughness. It was observed that cracks initiated and grew in the FiberForm when a critical stress limit was reached such that the carbon fibers separated from the binder. However, both for virgin and charred carbon ablators, crack initiation and growth occurred in the matrix (phenolic) phase. Both virgin and charred carbon ablators showed greater strength values compared with FiberForm samples, confirming that the presence of the porous matrix helps in absorbing the fracture energy.

  11. Percutaneous tumor ablation in medical radiology

    Energy Technology Data Exchange (ETDEWEB)

    Vogl, T.J.; Mack, M.G. [University Hospital Frankfurt Univ. (Germany). Inst. for Diagnostic and Interventional Radiology; Helmberger, T.K. [Klinikum Bogenhausen, Academic Teaching Hospital of the Technical Univ. Munich (Germany). Dept. for Diagnostic and Interventional Radiology and Nuclear Medicine; Reiser, M.F. (eds.) [University Hospitals - Grosshadern and Innenstadt Munich Univ. (Germany). Dept. of Clinical Radiology

    2008-07-01

    Thermal ablation has become an integral part of oncology, especially in the field of interventional oncology. This very comprehensive book encompasses the different technologies employed in thermal ablation, its indications and the results achieved in various clinical conditions. The first part of the book clearly explains the basics of thermal ablative techniques such as laser-induced thermotherapy, radiofrequency ablation, microwave ablation, cryotherapy, and localized tumor therapy. The latest developments in the application of minimally invasive therapies in localized neoplastic disease are demonstrated. In the main part of the book, techniques of guiding the applicators to the target structures by use of different imaging tools such as ultrasound, computed tomography and magnetic resonance imaging are discussed. The results are presented for a variety of clinical indications, including liver and lung tumors and metastases and some rather rare conditions involving the kidney, the head and neck, the prostate, and soft tissue structures. A large number of acknowledged experts have contributed to the book, which benefits from a lucid structure and excellent images. (orig.)

  12. Percutaneous tumor ablation in medical radiology

    International Nuclear Information System (INIS)

    Vogl, T.J.; Mack, M.G.; Helmberger, T.K.; Reiser, M.F.

    2008-01-01

    Thermal ablation has become an integral part of oncology, especially in the field of interventional oncology. This very comprehensive book encompasses the different technologies employed in thermal ablation, its indications and the results achieved in various clinical conditions. The first part of the book clearly explains the basics of thermal ablative techniques such as laser-induced thermotherapy, radiofrequency ablation, microwave ablation, cryotherapy, and localized tumor therapy. The latest developments in the application of minimally invasive therapies in localized neoplastic disease are demonstrated. In the main part of the book, techniques of guiding the applicators to the target structures by use of different imaging tools such as ultrasound, computed tomography and magnetic resonance imaging are discussed. The results are presented for a variety of clinical indications, including liver and lung tumors and metastases and some rather rare conditions involving the kidney, the head and neck, the prostate, and soft tissue structures. A large number of acknowledged experts have contributed to the book, which benefits from a lucid structure and excellent images. (orig.)

  13. Dynamic behaviors of laser ablated Si particles

    International Nuclear Information System (INIS)

    Ohyanagi, T.; Murakami, K.; Miyashita, A.; Yoda, O.

    1995-01-01

    The dynamics of laser-ablated Si particles produced by laser ablation have been investigated by time-and-space resolved X-ray absorption spectroscopy in a time scale ranging from 0 ns to 120 ns with a time resolution of 10 ns. Neutral and charged particles are observed through all X-ray absorption spectra. Assignments of transitions from 2s and 2p initial states to higher Rydberg states of Si atom and ions are achieved, and we experimentally determine the L II,III absorption edges of neutral Si atom (Si 0 ) and Si + , Si 2+ , Si 3+ and Si 4+ ions. The main ablated particles are found to be Si atom and Si ions in the initial stage of 0 ns to 120 ns. The relative amounts depend strongly on times and laser energy densities. We find that the spatial distributions of particles produced by laser ablation are changed with supersonic helium gas bombardment, but no cluster formation takes place. This suggests that a higher-density region of helium gas is formed at the top of the plume of ablated particles, and free expansion of particles is restrained by this helium cloud, and that it takes more than 120 ns to form Si clusters. (author)

  14. Influence of ablation wavelength and time on optical properties of laser ablated carbon dots

    Science.gov (United States)

    Isnaeni, Hanna, M. Yusrul; Pambudi, A. A.; Murdaka, F. H.

    2017-01-01

    Carbon dots, which are unique and applicable materials, have been produced using many techniques. In this work, we have fabricated carbon dots made of coconut fiber using laser ablation technique. The purpose of this work is to evaluate two ablation parameters, which are ablation wavelength and ablation time. We used pulsed laser from Nd:YAG laser with emit wavelength at 355 nm, 532 nm and 1064 nm. We varied ablation time one hour and two hours. Photoluminescence and time-resolved photoluminescence setup were used to study the optical properties of fabricated carbon dots. In general, fabricated carbon dots emit bluish green color emission upon excitation by blue laser. We found that carbon dots fabricated using 1064 nm laser produced the highest carbon dots emission among other samples. The peak wavelength of carbon dots emission is between 495 nm until 505 nm, which gives bluish green color emission. Two hours fabricated carbon dots gave four times higher emission than one hour fabricated carbon dot. More emission intensity of carbon dots means more carbon dots nanoparticles were fabricated during laser ablation process. In addition, we also measured electron dynamics of carbon dots using time-resolved photoluminescence. We found that sample with higher emission has longer electron decay time. Our finding gives optimum condition of carbon dots fabrication from coconut fiber using laser ablation technique. Moreover, fabricated carbon dots are non-toxic nanoparticles that can be applied for health, bio-tagging and medical applications.

  15. Bilateral cornual abscess after endometrial ablation following Essure sterilization.

    NARCIS (Netherlands)

    Jansen, N.E.; Vleugels, M.P.; Kluivers, K.B.; Vierhout, M.E.

    2007-01-01

    Endometrial ablation is used extensively to treat dysfunctional bleeding. Since the introduction of Essure tubal sterilization, this permanent contraception method has been widely used. Both endometrial ablation and Essure sterilization are procedures reported to have only a few complications. We

  16. Femtosecond laser ablation of carbon reinforced polymers

    International Nuclear Information System (INIS)

    Moreno, P.; Mendez, C.; Garcia, A.; Arias, I.; Roso, L.

    2006-01-01

    Interaction of intense ultrashort laser pulses (120 fs at 795 nm) with polymer based composites has been investigated. We have found that carbon filled polymers exhibit different ultrafast ablation behaviour depending on whether the filling material is carbon black or carbon fiber and on the polymer matrix itself. The shape and dimensions of the filling material are responsible for some geometrical bad quality effects in the entrance and inner surfaces of drilled microholes. We give an explanation for these non-quality effects in terms of fundamentals of ultrafast ablation process, specifically threshold laser fluences and material removal paths. Since carbon fiber reinforced polymers seemed particularly concerned, this could prevent the use of ultrafast ablation for microprocessing purposes of some of these materials

  17. Effects of endocardial microwave energy ablation

    Directory of Open Access Journals (Sweden)

    Vicente Climent

    2005-07-01

    Full Text Available Until recently the treatment of atrial fibrillation (AF consisted primarily of palliation, mostly in the form of pharmacological intervention. However because of recent advances in nonpharmacologic therapies, the current expectation of patients and referring physicians is that AF will be cured, rather than palliated. In recent years there has been a rapid expansion in the availability and variety of energy sources and devices for ablation. One of these energies, microwave, has been applied clinically only in the last few years, and may be a promising technique that is potentially capable of treating a wide range of ventricular and supraventricular arrhythmias. The purpose of this study was to review microwave energy ablation in surgical treatment of AF with special interest in histology and ultrastructure of lesions produced by this endocardial ablation procedure.

  18. Numerical Modeling of Ablation Heat Transfer

    Science.gov (United States)

    Ewing, Mark E.; Laker, Travis S.; Walker, David T.

    2013-01-01

    A unique numerical method has been developed for solving one-dimensional ablation heat transfer problems. This paper provides a comprehensive description of the method, along with detailed derivations of the governing equations. This methodology supports solutions for traditional ablation modeling including such effects as heat transfer, material decomposition, pyrolysis gas permeation and heat exchange, and thermochemical surface erosion. The numerical scheme utilizes a control-volume approach with a variable grid to account for surface movement. This method directly supports implementation of nontraditional models such as material swelling and mechanical erosion, extending capabilities for modeling complex ablation phenomena. Verifications of the numerical implementation are provided using analytical solutions, code comparisons, and the method of manufactured solutions. These verifications are used to demonstrate solution accuracy and proper error convergence rates. A simple demonstration of a mechanical erosion (spallation) model is also provided to illustrate the unique capabilities of the method.

  19. Advances in Imaging for Atrial Fibrillation Ablation

    International Nuclear Information System (INIS)

    D'Silva, A.; Wright, M.; Wright, M.

    2011-01-01

    Over the last fifteen years, our understanding of the pathophysiology of atrial fibrillation (AF) has paved the way for ablation to be utilized as an effective treatment option. With the aim of gaining more detailed anatomical representation, advances have been made using various imaging modalities, both before and during the ablation procedure, in planning and execution. Options have flourished from procedural fluoroscopy, electro anatomic mapping systems, pre procedural computed tomography (CT), magnetic resonance imaging (MRI), ultrasound, and combinations of these technologies. Exciting work is underway in an effort to allow the electro physiologist to assess scar formation in real time. One advantage would be to lessen the learning curve for what are very complex procedures. The hope of these developments is to improve the likelihood of a successful ablation procedure and to allow more patients access to this treatment

  20. Image-Guided Spinal Ablation: A Review

    Energy Technology Data Exchange (ETDEWEB)

    Tsoumakidou, Georgia, E-mail: gtsoumakidou@yahoo.com; Koch, Guillaume, E-mail: guillaume.koch@chru-strasbourg.fr; Caudrelier, Jean, E-mail: jean.caudrelier@chru-strasbourg.fr; Garnon, Julien, E-mail: julien.garnon@chru-strasbourg.fr; Cazzato, Roberto Luigi, E-mail: roberto-luigi.cazzato@chru-strasbourg.fr; Edalat, Faramarz, E-mail: faramarz.edalat@gmail.com; Gangi, Afshin, E-mail: gangi@unistra.fr [Strasbourg University Hospital (France)

    2016-09-15

    The image-guided thermal ablation procedures can be used to treat a variety of benign and malignant spinal tumours. Small size osteoid osteoma can be treated with laser or radiofrequency. Larger tumours (osteoblastoma, aneurysmal bone cyst and metastasis) can be addressed with radiofrequency or cryoablation. Results on the literature of spinal microwave ablation are scarce, and thus it should be used with caution. A distinct advantage of cryoablation is the ability to monitor the ice-ball by intermittent CT or MRI. The different thermal insulation, temperature and electrophysiological monitoring techniques should be applied. Cautious pre-procedural planning and intermittent intra-procedural monitoring of the ablation zone can help reduce neural complications. Tumour histology, patient clinical-functional status and life-expectancy should define the most efficient and least disabling treatment option.

  1. Deep Dive Topic: Choosing between ablators

    Energy Technology Data Exchange (ETDEWEB)

    Hurricane, O. A. [Lawrence Livermore National Lab. (LLNL), Livermore, CA (United States); Thomas, C. [Lawrence Livermore National Lab. (LLNL), Livermore, CA (United States); Olson, R. [Lawrence Livermore National Lab. (LLNL), Livermore, CA (United States)

    2015-07-14

    Recent data on implosions using identical hohlraums and very similar laser drives underscores the conundrum of making a clear choice of one ablator over another. Table I shows a comparison of Be and CH in a nominal length, gold, 575 μm-diameter, 1.6 mg/cc He gas-fill hohlraum while Table II shows a comparison of undoped HDC and CH in a +700 length, gold, 575 μm diameter, 1.6 mg/cc He gas fill hohlraum. As can be seen in the tables, the net integrated fusion performance of these ablators is the same to within error bars. In the case of the undoped HDC and CH ablators, the hot spot shapes of the implosions were nearly indistinguishable for the experiments listed in Table II.

  2. Kinetic depletion model for pellet ablation

    International Nuclear Information System (INIS)

    Kuteev, Boris V.

    2001-11-01

    A kinetic model for depletion effect, which determines pellet ablation when the pellet passes a rational magnetic surface, is formulated. The model predicts a moderate decrease of the ablation rate compared with the earlier considered monoenergy versions [1, 2]. For typical T-10 conditions the ablation rate reduces by a reactor of 2.5 when the 1-mm pellet penetrates through the plasma center. A substantial deceleration of pellets -about 15% per centimeter of low shire rational q region; is predicted. Penetration for Low Field Side and High Field Side injections is considered taking into account modification of the electron distribution function by toroidal magnetic field. It is shown that Shafranov shift and toroidal effects yield the penetration length for HFS injection higher by a factor of 1.5. This fact should be taken into account when plasma-shielding effects on penetration are considered. (author)

  3. In vivo study of necrosis on the liver tissue of Wistar rats: a combination of photodynamic therapy and carbon dioxide laser ablation

    International Nuclear Information System (INIS)

    Rego, R F; Nicolodelli, G; Bagnato, V S; Araujo, M T; Tirapelli, L F; Araujo-Moreira, F M

    2013-01-01

    Photodynamic therapy (PDT) is known to be limited to applications in large volume tumors due to its limited penetration. Therefore, a combination of PDT and carbon dioxide (CO 2 ) laser ablation may constitute a potential protocol to destroy bulk tumors because it involves an association of these two techniques allowing the removal of visible lesions with a high selectivity of destruction of remnant tumors. The main aim of this study is to investigate the most appropriate procedure to combine use of a CO 2 laser and PDT on livers of healthy rats, and to analyze different techniques of this treatment using three types of photosensitizers (PSs). Forty eight animals were separated to form six groups: (1) only CO 2 laser ablation, (2) drug and CO 2 laser ablation, (3) only PDT, (4) drug and light (PDT) followed by CO 2 laser ablation, (5) ablated with CO 2 laser followed by PDT, and (6) drug followed by CO 2 laser ablation and light. For each group, three types of photosensitization were used: topical 5-aminolevulinic acid (ALA), intravenous ALA and intravenous Photogem ® . Thirty hours after the treatments, the animals were sacrificed and the livers removed. The depth of necrosis was analyzed and measured, considering microscopic and macroscopic aspects. The results show that the effects of the PDT were considerably enhanced when combined with CO 2 laser ablation, especially when the PDT was performed before the CO 2 laser ablation. (paper)

  4. Effect of Radiofrequency Endometrial Ablation on Dysmenorrhea.

    Science.gov (United States)

    Wyatt, Sabrina N; Banahan, Taylor; Tang, Ying; Nadendla, Kavita; Szychowski, Jeff M; Jenkins, Todd R

    To examine rates of dysmenorrhea after radiofrequency endometrial ablation in patients with and without known dysmenorrhea symptoms prior to the procedure in a diverse population. Retrospective cohort study (Canadian Task Force classification II-2). Academic gynecology practice. A total of 307 women underwent endometrial ablation between 2007 and 2013 at our institution. Patients who had preoperative and postoperative pain symptom assessments as well as a description of pain timing recorded were included in our analysis. Exclusion criteria were age dysmenorrhea was evaluated. Demographic information and other outcome variables were used to evaluate factors associated with resolution of dysmenorrhea. A total of 307 patients who underwent radiofrequency endometrial ablation were identified. After exclusions, 296 charts were examined, and 144 patients met our enrollment criteria. The mean age of the study cohort was 45.4 ± 6.2 years; 57 patients (40%) were African American, 16 (11%) had a body mass index (BMI) > 40, and 41 (29%) were of normal weight. Preoperative dysmenorrhea was reported by 100 patients (69%); 48 of these patients (48%) experienced resolution of symptoms postoperatively. Only 3 of the 44 patients (7%) without preoperative dysmenorrhea reported new-onset dysmenorrhea postoperatively. Significantly fewer patients had dysmenorrhea after compared to before radiofrequency ablation (55 of 144 [38%] vs 100 of 144 [69%]; p dysmenorrhea after ablation was associated with reduction in bleeding volume (p = .048) but not with a reduction in frequency of bleeding (p = .12). Approximately one-half of women who undergo radiofrequency endometrial ablation to treat heavy menstrual bleeding who also have preoperative dysmenorrhea exhibit documented pain resolution after the procedure. Resolution of dysmenorrhea is more likely if menstrual flow volume is decreased postprocedure. Copyright © 2016 AAGL. Published by Elsevier Inc. All rights reserved.

  5. Temperature-controlled irrigated tip radiofrequency catheter ablation

    DEFF Research Database (Denmark)

    Petersen, H H; Chen, X; Pietersen, Adrian

    1998-01-01

    INTRODUCTION: In patients with ventricular tachycardias due to structural heart disease, catheter ablation cures radiofrequency ablation. Irrigated tip radiofrequency ablation using power control and high infusion rates enlarges lesion......: We conclude that temperature-controlled radiofrequency ablation with irrigated tip catheters using low target temperature and low infusion rate enlarges lesion size without increasing the incidence of cratering and reduces coagulum formation of the tip....

  6. Subcellular analysis by laser ablation electrospray ionization mass spectrometry

    Science.gov (United States)

    Vertes, Akos; Stolee, Jessica A; Shrestha, Bindesh

    2014-12-02

    In various embodiments, a method of laser ablation electrospray ionization mass spectrometry (LAESI-MS) may generally comprise micro-dissecting a cell comprising at least one of a cell wall and a cell membrane to expose at least one subcellular component therein, ablating the at least one subcellular component by an infrared laser pulse to form an ablation plume, intercepting the ablation plume by an electrospray plume to form ions, and detecting the ions by mass spectrometry.

  7. Thermal Ablation for Benign Thyroid Nodules: Radiofrequency and Laser

    Energy Technology Data Exchange (ETDEWEB)

    Baek, Jung Hwan; Lee, Jeong Hyun [University of Ulsan College of Medicine, Asan Medical Center, Seoul (Korea, Republic of); Valcavi, Roberto [Endocrinology Division and Thyroid Disease Center, Arcispedale Santa Maria Nuova, Reggio Emilia (Italy); Pacella, Claudio M. [Diagnostic Imaging and Interventional Radiology Department, Ospedale Regina Apostolorum, Albano Laziale-Rome (IT); Rhim, Hyun Chul [Samsung Medical Center, Sungkyunkwan University School of Medicine, Seoul (Korea, Republic of); Na, Dong Kyu [Human Medical Imaging and Intervention Center, Seoul (Korea, Republic of)

    2011-10-15

    Although ethanol ablation has been successfully used to treat cystic thyroid nodules, this procedure is less effective when the thyroid nodules are solid. Radiofrequency (RF) ablation, a newer procedure used to treat malignant liver tumors, has been valuable in the treatment of benign thyroid nodules regardless of the extent of the solid component. This article reviews the basic physics, techniques, applications, results, and complications of thyroid RF ablation, in comparison to laser ablation.

  8. Chimeric forms of furin and TGN38 are transported with the plasma membrane in the trans-Golgi network via distinct endosomal pathways.

    Science.gov (United States)

    Mallet, W G; Maxfield, F R

    1999-07-26

    Furin and TGN38 are menbrane proteins that cycle between the plasma membrane and the trans-Golgi network (TGN), each maintaining a predominant distribution in the TGN. We have used chimeric proteins with an extracellular Tac domain and the cytoplasmic domain of TGN38 or furin to study the trafficking of these proteins in endosomes. Previously, we demonstrated that the postendocytic trafficking of Tac-TGN38 to the TGN is via the endocytic recycling pathway (Ghosh, R.N.,W.G. Mallet,T.T. Soe,T.E.McGraw, and F.R. Maxfield.1998.J. Cell Biol.142:923-936). Here we show that internalized Tac-furin is delivered to the TGN through late endosomes, bypassing the endocytic recycling compartment. The transport of Tac-furin from late endosomes to the TGN appears to proceed via an efficient, single-pass mechanism. Delivery of Tac-furin but not Tac-TGN38 to the TGN is blocked by nocodazole, and the two pathways are also differentially affected by wortmannin. These studies demonstrate the existence of two independentpathways for endosomal transport of proteins to the TGN from the plasma membrane.

  9. Increased expression of endosomal members of toll-like receptor family abrogates wound healing in patients with type 2 diabetes mellitus.

    Science.gov (United States)

    Singh, Kanhaiya; Agrawal, Neeraj K; Gupta, Sanjeev K; Mohan, Gyanendra; Chaturvedi, Sunanda; Singh, Kiran

    2016-10-01

    The inflammatory phase of wound healing cascade is an important determinant of the fate of the wound. Acute inflammation is necessary to initiate proper wound healing, while chronic inflammation abrogates wound healing. Different endosomal members of toll-like receptor (TLR) family initiate inflammatory signalling via a range of different inflammatory mediators such as interferons, internal tissue damaged-associated molecular patterns (DAMPs) and hyperactive effector T cells. Sustained signalling of TLR9 and TLR7 contributes to chronic inflammation by activating the plasmacytoid dendritic cells. Diabetic wounds are also characterised by sustained inflammatory phase. The objective of this study was to analyse the differential expression of endosomal TLRs in human diabetic wounds compared with control wounds. We analysed the differential expression of TLR7 and TLR9 both at transcriptional and translational levels in wounds of 84 patients with type 2 diabetes mellitus (T2DM) and 6 control subjects without diabetes using quantitative real-time polymerase chain reaction (RT-PCR), western blot and immunohistochemistry. TLR7 and TLR9 were significantly up-regulated in wounds of the patients with T2DM compared with the controls and were dependent on the infection status of the diabetic wounds, and wounds with microbial infection exhibited lower expression levels of endosomal TLRs. Altered endosomal TLR expression in T2DM subjects might be associated with wound healing impairment. © 2015 Medicalhelplines.com Inc and John Wiley & Sons Ltd.

  10. Strategies for the Activation and Release of the Membranolytic Peptide Melittin from Liposomes Using Endosomal pH as a Trigger

    NARCIS (Netherlands)

    Oude Blenke, E.; Sleszynska, M.; Evers, M. J W; Storm, G.; Martin, N. I.; Mastrobattista, E.

    2017-01-01

    Endosomolytic peptides are often coupled to drug delivery systems to enhance endosomal escape, which is crucial for the delivery of macromolecular drugs that are vulnerable to degradation in the endolysosomal pathway. Melittin is a 26 amino acid peptide derived from bee venom that has a very high

  11. High throughput solar cell ablation system

    Science.gov (United States)

    Harley, Gabriel; Pass, Thomas; Cousins, Peter John; Viatella, John

    2012-09-11

    A solar cell is formed using a solar cell ablation system. The ablation system includes a single laser source and several laser scanners. The laser scanners include a master laser scanner, with the rest of the laser scanners being slaved to the master laser scanner. A laser beam from the laser source is split into several laser beams, with the laser beams being scanned onto corresponding wafers using the laser scanners in accordance with one or more patterns. The laser beams may be scanned on the wafers using the same or different power levels of the laser source.

  12. Endosomal Escape and Delivery of CRISPR/Cas9 Genome Editing Machinery Enabled by Nanoscale Zeolitic Imidazolate Framework

    KAUST Repository

    Alsaiari, Shahad K.

    2017-12-22

    CRISPR/Cas9 is a combined protein (Cas9) and an engineered single guide RNA (sgRNA) genome editing platform that offers revolutionary solutions to genetic diseases. It has, however, a double delivery problem owning to the large protein size and the highly charged RNA component. In this work, we report the first example of CRISPR/Cas9 encapsulated by nanoscale zeolitic imidazole frameworks (ZIFs) with a loading efficiency of 17% and enhanced endosomal escape promoted by the protonated imidazole moieties. The gene editing potential of CRISPR/Cas9 encapsulated by ZIF-8 (CC-ZIFs) is further verified by knocking down the gene expression of green fluorescent protein by 37% over 4 days employing CRISPR/Cas9 machinery. The nanoscale CC-ZIFs are biocompatible and easily scaled-up offering excellent loading capacity and controlled co-delivery of intact Cas9 protein and sgRNA.

  13. Endosomal Escape and Delivery of CRISPR/Cas9 Genome Editing Machinery Enabled by Nanoscale Zeolitic Imidazolate Framework

    KAUST Repository

    Alsaiari, Shahad K.; Patil, Sachin; Alyami, Mram Z.; Alamoudi, Kholod; Aleisa, Fajr A; Merzaban, Jasmeen; Li, Mo; Khashab, Niveen M.

    2017-01-01

    CRISPR/Cas9 is a combined protein (Cas9) and an engineered single guide RNA (sgRNA) genome editing platform that offers revolutionary solutions to genetic diseases. It has, however, a double delivery problem owning to the large protein size and the highly charged RNA component. In this work, we report the first example of CRISPR/Cas9 encapsulated by nanoscale zeolitic imidazole frameworks (ZIFs) with a loading efficiency of 17% and enhanced endosomal escape promoted by the protonated imidazole moieties. The gene editing potential of CRISPR/Cas9 encapsulated by ZIF-8 (CC-ZIFs) is further verified by knocking down the gene expression of green fluorescent protein by 37% over 4 days employing CRISPR/Cas9 machinery. The nanoscale CC-ZIFs are biocompatible and easily scaled-up offering excellent loading capacity and controlled co-delivery of intact Cas9 protein and sgRNA.

  14. Concerted regulation of npc2 binding to endosomal/lysosomal membranes by bis(monoacylglycero)phosphate and sphingomyelin

    DEFF Research Database (Denmark)

    Enkavi, Giray; Mikkolainen, Heikki; Güngör, Burçin

    2017-01-01

    remained elusive. Here, based on an extensive set of atomistic simulations and free energy calculations, we clarify the mechanism and energetics of npc2-membrane binding and characterize the roles of physiologically relevant key lipids associated with the binding process. Our results capture in atomistic......Niemann-Pick Protein C2 (npc2) is a small soluble protein critical for cholesterol transport within and from the lysosome and the late endosome. Intriguingly, npc2-mediated cholesterol transport has been shown to be modulated by lipids, yet the molecular mechanism of npc2-membrane interactions has......). This mode is associated with cholesterol uptake and release. On the other hand, the second mode (Supine) places the cholesterol binding pocket away from the membrane surface, but has overall higher membrane binding affinity. We determined that bis(monoacylglycero)phosphate (bmp) is specifically required...

  15. Constitutively internalized dopamine transporter is targeted to late endosomes and lysosomal degradation in heterologous cell lines and dopaminergic neurons

    DEFF Research Database (Denmark)

    Eriksen, Jacob; Madsen, Kenneth; Vægter, Christian Bjerggaard

    and amphetamine, a substrate of the DAT. In antibody feeding experiments we observed that Tac-DAT was constitutively internalized faster than Tac alone and using an ELISA based assay we could quantify time-dependent intracellular accumulation of the transporter. Incubation with inhibitors of lysosomal degradation...... (leupeptin, chloroquine, or ammonium chloride) increased the amount of transporter accumulated intracellularly over time, suggesting that constitutively endocytosed transporter was targeted to lysosomal degradation. This was further supported by expression of Tac-DAT in the immortalized dopaminergic cell...... dopaminergic neurons and visualized the DAT directly in the neurons using the fluorescent cocaine analog JHC 1-064. These data showed pronounced colocalization upon constitutive internalization with Lysotracker, a late endosomal/lysosomal marker; however only little co-lolization was observed with Alexa488...

  16. Efficacy and satisfaction rate comparing endometrial ablation by rollerball electrocoagulation to uterine balloon thermal ablation in a randomised controlled trial.

    NARCIS (Netherlands)

    Zon-Rabelink, I.A.A. van; Vleugels, M.P.; Merkus, J.M.W.M.; Graaf, R.M. de

    2004-01-01

    OBJECTIVE: To compare two methods of endometrial ablation, hysteroscopic rollerball electrocoagulation (RBE) and non-hysteroscopic uterine balloon thermal ablation (Thermachoice trade mark ), regarding efficacy for reducing dysfunctional uterine bleeding and patients satisfaction rate. METHODS: A

  17. The case against endometrial ablation for treatment of heavy menstrual bleeding.

    Science.gov (United States)

    Louie, Michelle; Wright, Kelly; Siedhoff, Matthew

    2018-04-27

    Endometrial ablation is a common treatment for heavy menstrual bleeding, but serious limitations and long-term complications exist. Our purpose is to summarize the use of endometrial ablation devices, potential short-term and long-term complications, cost effectiveness, and quality of life in relation to alternative treatments. There is insufficient evidence to strongly recommend one endometrial ablation device over another. Providers should consider and discuss with their patients, complications including risk of future pregnancy, endometrial cancer, and hysterectomy for continued bleeding or pain. Patient selection is key to reducing postablation pain and failure; patients with a history of tubal ligation and dysmenorrhea should consider alternative treatments. All patients should also be counseled that the levonorgestrel intrauterine device is a cost-effective alternative with higher quality of life and fewer complications. Hysterectomy is definitive treatment with higher quality of life and fewer complications. Although endometrial ablation can offer adequate symptom control for patients who have failed medical therapy, desire uterine preservation, or who are high-risk surgical candidates, patients should be appropriately selected and counseled regarding the potential for treatment failure and long-term complications.

  18. Conventional versus 3-D Echocardiography to Predict Arrhythmia Recurrence After Atrial Fibrillation Ablation.

    Science.gov (United States)

    Bossard, Matthias; Knecht, Sven; Aeschbacher, Stefanie; Buechel, Ronny R; Hochgruber, Thomas; Zimmermann, Andreas J; Kessel-Schaefer, Arnheid; Stephan, Frank-Peter; Völlmin, Gian; Pradella, Maurice; Sticherling, Christian; Osswald, Stefan; Kaufmann, Beat A; Conen, David; Kühne, Michael

    2017-06-01

    Arrhythmia recurrence after atrial fibrillation (AF) ablation remains high and requires repeat interventions in a substantial number of patients. We assessed the value of conventional and 3-D echocardiography to predict AF recurrence. Consecutive patients undergoing AF ablation by means of pulmonary vein isolation were included in a prospective registry. Echocardiograms were obtained prior to the ablation procedure, and analyzed offline in a standardized manner, including 3-D left atrial (LA) volumetry and determination of LA function and sphericity. The primary endpoint, AF recurrence (>30 seconds) between 3 to 12 months after AF ablation, was independently adjudicated. We included 276 patients (73% male, mean age 59.9 ± 9.9 years). Paroxysmal and persistent AF were present in 178 (64%) and 98 (36%) patients, respectively. Mean left ventricular ejection fraction and indexed LA volume in 3-D (LAVI) were 52 ± 12% and 42 ± 13 mL/m 2 , respectively. AF recurrence was observed in 110 (40%) patients after a single procedure. Median (interquartile range) time to AF recurrence was 123 (92; 236) days. In multivariable Cox regression models, the only predictors for AF recurrence were the minimal, maximal, and indexed 3-D LA volumes, P = 0.024, P = 0.016, and P = 0.014, respectively. Quartile specific analysis of 3-D LAVI showed an HR of 1.885 (95%CI 1.066-3.334; P for trend = 0.015) for the highest compared to the lowest quartile. Our results show the important role of LA volume for the long-term freedom from arrhythmia after AF ablation. These data also highlight the potential of 3-D echocardiography in this context and may facilitate patient selection for AF ablation. © 2017 Wiley Periodicals, Inc.

  19. Endoscopic radiofrequency ablation therapy for the prevention of esophageal cancer in Barrett’s esophagus

    Directory of Open Access Journals (Sweden)

    Ha NH

    2015-07-01

    Full Text Available Ngoc Hoang Ha, Richard Hummel, David I WatsonDepartment of Surgery, Flinders University, Flinders Medical Centre, Bedford Park, Adelaide, South Australia, AustraliaAbstract: Barrett’s esophagus is the only known precursor lesion for esophageal adenocarcinoma. Previous studies have shown that a variety of methods can be applied to destroy Barrett’s esophagus epithelium, and healing with a new esophageal squamous epithelium usually occurs following ablation. Radiofrequency ablation (RFA is a relatively new endoscopic technique. It has been claimed that ablation using RFA reduces the risk of cancer progression. RFA is usually easy to apply and is associated with a low risk of morbidity. It achieves complete eradication of (non dysplastic Barrett’s esophagus in most individuals, and the risk of progression to higher grades of dysplasia or cancer is reduced after RFA, although not completely eliminated. Limitations include recurrence of Barrett’s esophagus in up to one-third of individuals, a risk of “buried islands” of Barrett’s esophagus remaining below the regenerated mucosa, and uncertainty about the biological behavior of the new squamous epithelium after RFA. Current evidence supports the use of RFA in individuals with high-grade dysplasia in Barrett’s esophagus, and early stage (T1a intramucosal cancer, and select individuals with low-grade dysplasia. As accurate diagnosis of low-grade dysplasia remains difficult outside expert centers, it is probably premature to recommend routine RFA for all patients diagnosed with low-grade dysplasia in the community, despite the favorable outcomes from one randomized trial. Furthermore, long-term outcomes following ablation remain uncertain, and ongoing endoscopy surveillance is still required after RFA as progression to cancer remains a possibility. Outcomes from large studies with long-term follow-up are needed to definitively confirm that RFA ablation can reliably prevent cancer

  20. The endosomal recycling of FgSnc1 by FgSnx41-FgSnx4 heterodimer is essential for polarized growth and pathogenicity in Fusarium graminearum.

    Science.gov (United States)

    Zheng, Wenhui; Lin, Yahong; Fang, Wenqin; Zhao, Xu; Lou, Yi; Wang, Guanghui; Zheng, Huawei; Liang, Qifu; Abubakar, Yakubu Saddeeq; Olsson, Stefan; Zhou, Jie; Wang, Zonghua

    2018-04-20

    Endosomal sorting machineries regulate the transport of their cargoes among intracellular compartments. However, the molecular nature of such intracellular trafficking processes in pathogenic fungal development and pathogenicity remains unclear. Here, we dissect the roles and molecular mechanisms of two sorting nexin proteins and their cargoes in endosomal recycling in Fusarium graminearum using high-resolution microscopy and high-throughput co-immunoprecipitation strategies. We show that the sorting nexins, FgSnx41 and FgSnx4, interact with each other and assemble into a functionally interdependent heterodimer through their respective BAR domains. Further analyses demonstrate that the dimer localizes to the early endosomal membrane and coordinates endosomal sorting. The small GTPase FgRab5 regulates the correct localization of FgSnx41-FgSnx4 and is consequently required for its trafficking function. The protein FgSnc1 is a cargo of FgSnx41-FgSnx4 and regulates the fusion of secreted vesicles with the fungal growing apex and plasma membrane. In the absence of FgSnx41 or FgSnx4, FgSnc1 is mis-sorted and degraded in the vacuole, and null deletion of either component causes defects in the fungal polarized growth and virulence. Overall, for the first time, our results reveal the mechanism of FgSnc1 endosomal recycling by FgSnx41-FgSnx4 heterodimer which is essential for polarized growth and pathogenicity in F. graminearum. © 2018 The Authors. New Phytologist © 2018 New Phytologist Trust.

  1. Gαs regulates Glucagon-Like Peptide 1 Receptor-mediated cyclic AMP generation at Rab5 endosomal compartment.

    Science.gov (United States)

    Girada, Shravan Babu; Kuna, Ramya S; Bele, Shilpak; Zhu, Zhimeng; Chakravarthi, N R; DiMarchi, Richard D; Mitra, Prasenjit

    2017-10-01

    Upon activation, G protein coupled receptors (GPCRs) associate with heterotrimeric G proteins at the plasma membrane to initiate second messenger signaling. Subsequently, the activated receptor experiences desensitization, internalization, and recycling back to the plasma membrane, or it undergoes lysosomal degradation. Recent reports highlight specific cases of persistent cyclic AMP generation by internalized GPCRs, although the functional significance and mechanistic details remain to be defined. Cyclic AMP generation from internalized Glucagon-Like Peptide-1 Receptor (GLP-1R) has previously been reported from our laboratory. This study aimed at deciphering the molecular mechanism by which internalized GLP-R supports sustained cyclic AMP generation upon receptor activation in pancreatic beta cells. We studied the time course of cyclic AMP generation following GLP-1R activation with particular emphasis on defining the location where cyclic AMP is generated. Detection involved a novel GLP-1 conjugate coupled with immunofluorescence using specific endosomal markers. Finally, we employed co-immunoprecipitation as well as immunofluorescence to assess the protein-protein interactions that regulate GLP-1R mediated cyclic AMP generation at endosomes. Our data reveal that prolonged association of G protein α subunit Gαs with activated GLP-1R contributed to sustained cyclic AMP generation at Rab 5 endosomal compartment. The findings provide the mechanism of endosomal cyclic AMP generation following GLP-1R activation. We identified the specific compartment that serves as an organizing center to generate endosomal cyclic AMP by internalized activated receptor complex. Copyright © 2017 The Authors. Published by Elsevier GmbH.. All rights reserved.

  2. Laser ablation of posterior urethral valves by fetal cystoscopy.

    Science.gov (United States)

    Martínez, José María; Masoller, Narcis; Devlieger, Roland; Passchyn, Esther; Gómez, Olga; Rodo, Joan; Deprest, Jan A; Gratacós, Eduard

    2015-01-01

    To report the results of fetal cystoscopic laser ablation of posterior urethral valves (PUV) in a consecutive series in two referral centers. Twenty pregnant women with a presumptive isolated PUV were treated with fetal cystoscopy under local anesthesia. Identification and fulguration of the PUV by one or several firing-contacts with diode laser were attempted. Perinatal and long-term outcomes were prospectively recorded. The median gestational age at procedure was 18.1 weeks (range 15.0-25.6), and median operation time was 24 min (range 15-40). Access to the urethra was achieved in 19/20 (95%) cases, and postoperative, normalization of bladder size and amniotic fluid was observed in 16/20 (80%). Overall, there were 9 (45%) terminations of pregnancy and 11 women (55%) delivered a liveborn baby at a mean gestational age of 37.3 (29.1-40.2) weeks. No infants developed pulmonary hypoplasia and all were alive at 15-110 months. Eight (40% of all fetuses, 72.7% of newborns) had normal renal function and 3 (27.3%) had renal failure awaiting renal transplantation. Fetoscopic laser ablation for PUV can achieve bladder decompression and amniotic fluid normalization with a single procedure in selected cases with anyhydramnios. There is still a significant risk of progression to renal failure pre or postnatally. © 2014 S. Karger AG, Basel.

  3. Stereotactic Ablative Radiotherapy for Oligometastatic Disease in Liver

    Directory of Open Access Journals (Sweden)

    Myungsoo Kim

    2014-01-01

    Full Text Available Liver metastasis in solid tumors, including colorectal cancer, is the most frequent and lethal complication. The development of systemic therapy has led to prolonged survival. However, in selected patients with a finite number of discrete lesions in liver, defined as oligometastatic state, additional local therapies such as surgical resection, radiofrequency ablation, cryotherapy, and radiotherapy can lead to permanent local disease control and improve survival. Among these, an advance in radiation therapy made it possible to deliver high dose radiation to the tumor more accurately, without impairing the liver function. In recent years, the introduction of stereotactic ablative radiotherapy (SABR has offered even more intensive tumor dose escalation in a few fractions with reduced dose to the adjacent normal liver. Many studies have shown that SABR for oligometastases is effective and safe, with local control rates widely ranging from 50% to 100% at one or two years. And actuarial survival at one and two years has been reported ranging from 72% to 94% and from 30% to 62%, respectively, without severe toxicities. In this paper, we described the definition and technical aspects of SABR, clinical outcomes including efficacy and toxicity, and related parameters after SABR in liver oligometastases from colorectal cancer.

  4. A review of the safety aspects of radio frequency ablation

    Directory of Open Access Journals (Sweden)

    Abhishek Bhaskaran

    2015-09-01

    Full Text Available In light of recent reports showing high incidence of silent cerebral infarcts and organized atrial arrhythmias following radiofrequency (RF atrial fibrillation (AF ablation, a review of its safety aspects is timely. Serious complications do occur during supraventricular tachycardia (SVT ablations and knowledge of their incidence is important when deciding whether to proceed with ablation. Evidence is emerging for the probable role of prophylactic ischemic scar ablation to prevent VT. This might increase the number of procedures performed. Here we look at the various complications of RF ablation and also the methods to minimize them. Electronic database was searched for relevant articles from 1990 to 2015. With better awareness and technological advancements in RF ablation the incidence of complications has improved considerably. In AF ablation it has decreased from 6% to less than 4% comprising of vascular complications, cardiac tamponade, stroke, phrenic nerve injury, pulmonary vein stenosis, atrio-esophageal fistula (AEF and death. Safety of SVT ablation has also improved with less than 1% incidence of AV node injury in AVNRT ablation. In VT ablation the incidence of major complications was 5–11%, up to 3.4%, up to 1.8% and 4.1–8.8% in patients with structural heart disease, without structural heart disease, prophylactic ablations and epicardial ablations respectively. Vascular and pericardial complications dominated endocardial and epicardial VT ablations respectively. Up to 3% mortality and similar rates of tamponade were reported in endocardial VT ablation. Recent reports about the high incidence of asymptomatic cerebral embolism during AF ablation are concerning, warranting more research into its etiology and prevention.

  5. Ultrasound-guided percutaneous thermal ablation of hepatocellular carcinoma using microwave and radiofrequency ablation

    Energy Technology Data Exchange (ETDEWEB)

    Xu, H.-X.; Xie, X.-Y.; Lu, M.-D. E-mail: lumd@21cn.com; Chen, J.-W.; Yin, X.-Y.; Xu, Z.-F.; Liu, G.-J

    2004-01-01

    AIM: To investigate the therapeutic efficacy of thermal ablation for treatment of hepatocellular carcinoma (HCC) using microwave and radiofrequency (RF) energy application. MATERIALS AND METHODS: A total of 190 nodules in 97 patients (84 male, 13 female; mean age 53.4 years, range 24-74 years) with HCC were treated with microwave or RF ablation in the last 4 years. The applicators were introduced into the tumours under conscious analgesic sedation by intravenous administration of fentanyl citrate and droperidol and local anaesthesia in both thermal ablation procedures. The patients were then followed up with contrast-enhanced computed tomography (CT) to evaluate treatment response. Survival was analysed using the Kaplan-Meier method. RESULTS: Complete ablation was obtained in 92.6% (176/190) nodules. The complete ablation rates were 94.6% (106/112) in microwave ablation and 89.7% (70/78) in RF ablation. The complete ablation rates in tumours{<=}2.0, 2.1-3.9 and {>=}4.0 cm were 93.1, 93.8 and 86.4%, respectively. Local recurrence was found in 9.5% nodules and the rates in tumours{<=}2.0, 2.1-3.9 and {>=}4.0 cm in diameter were 3.4, 9.9 and 31.8%, respectively. In the follow-up period, 7.1% nodules ablated by microwave and 12.8% by RF presented local recurrence. The 1, 2 and 3-year distant recurrence-free survivals were 47.2, 34.9 and 31.0%, respectively. Estimated mean survival was 32 months, and 1, 2 and 3-year cumulative survivals were 75.6, 58.5, and 50.0%, respectively. One and 2 years survivals of Child-Pugh class A, B and C patients were 83.8 and 70.4%, 78.2 and 53.2%, 36.3 and 27.3%, respectively. CONCLUSION: Thermal ablation therapy by means of microwave and RF energy application is an effective and safe therapeutic technique for hepatocellular carcinoma. Large tumours can be completely ablated, but have a significantly higher risk of local recurrence at follow-up.

  6. Ultrasound-guided percutaneous thermal ablation of hepatocellular carcinoma using microwave and radiofrequency ablation

    International Nuclear Information System (INIS)

    Xu, H.-X.; Xie, X.-Y.; Lu, M.-D.; Chen, J.-W.; Yin, X.-Y.; Xu, Z.-F.; Liu, G.-J.

    2004-01-01

    AIM: To investigate the therapeutic efficacy of thermal ablation for treatment of hepatocellular carcinoma (HCC) using microwave and radiofrequency (RF) energy application. MATERIALS AND METHODS: A total of 190 nodules in 97 patients (84 male, 13 female; mean age 53.4 years, range 24-74 years) with HCC were treated with microwave or RF ablation in the last 4 years. The applicators were introduced into the tumours under conscious analgesic sedation by intravenous administration of fentanyl citrate and droperidol and local anaesthesia in both thermal ablation procedures. The patients were then followed up with contrast-enhanced computed tomography (CT) to evaluate treatment response. Survival was analysed using the Kaplan-Meier method. RESULTS: Complete ablation was obtained in 92.6% (176/190) nodules. The complete ablation rates were 94.6% (106/112) in microwave ablation and 89.7% (70/78) in RF ablation. The complete ablation rates in tumours≤2.0, 2.1-3.9 and ≥4.0 cm were 93.1, 93.8 and 86.4%, respectively. Local recurrence was found in 9.5% nodules and the rates in tumours≤2.0, 2.1-3.9 and ≥4.0 cm in diameter were 3.4, 9.9 and 31.8%, respectively. In the follow-up period, 7.1% nodules ablated by microwave and 12.8% by RF presented local recurrence. The 1, 2 and 3-year distant recurrence-free survivals were 47.2, 34.9 and 31.0%, respectively. Estimated mean survival was 32 months, and 1, 2 and 3-year cumulative survivals were 75.6, 58.5, and 50.0%, respectively. One and 2 years survivals of Child-Pugh class A, B and C patients were 83.8 and 70.4%, 78.2 and 53.2%, 36.3 and 27.3%, respectively. CONCLUSION: Thermal ablation therapy by means of microwave and RF energy application is an effective and safe therapeutic technique for hepatocellular carcinoma. Large tumours can be completely ablated, but have a significantly higher risk of local recurrence at follow-up

  7. Economic and clinical benefits of endometrial radiofrequency ablation compared with other ablation techniques in women with menorrhagia: a retrospective analysis with German health claims data

    Directory of Open Access Journals (Sweden)

    Bischoff-Everding C

    2016-01-01

    Full Text Available Christoph Bischoff-Everding,1 Ruediger Soeder,2 Benno Neukirch3 1HGC GesundheitsConsult GmbH, Duesseldorf, Germany; 2Gynecological Joint Practice, Mainz, Germany; 3Faculty of Health Care, Hochschule Niederrhein – University of Applied Sciences, Krefeld, Germany Objective: To evaluate the economic and clinical benefits of endometrial radiofrequency ablation (RFA compared with other ablation techniques for the treatment of menorrhagia.Methods: Using German health claims data, women meeting defined inclusion criteria for the intervention group (RFA were selected. A comparable control group (other endometrial ablations was established using propensity score matching. These two groups were compared during the quarter of treatment (QoT and a follow-up of 2 years for the following outcomes: costs during QoT and during follow-up, repeated menorrhagia diagnoses during follow-up and necessary retreatments during follow-up. Results: After performing propensity score matching, 50 cases could be allocated to the intervention group, while 38 were identified as control cases. Patients in the RFA group had 5% fewer repeat menorrhagia diagnoses (40% vs 45%; not significant and 5% fewer treatments associated with recurrent menorrhagia (6% vs 11%; not significant than cases in the control group. During the QoT, the RFA group incurred €578 additional costs (€2,068 vs €1,490; ns. However, during follow-up, the control group incurred €1,254 additional costs (€4,561 vs €5,815; ns, with medication, outpatient physician consultations, and hospitals costs being the main cost drivers. However, none of the results were statistically significant. Conclusion: Although RFA was more cost-intensive in the QoT compared with other endometrial ablation techniques, an average total savings of €676 was generated during the follow-up period. While having evidence that RFA is clinically equivalent to other endometrial ablation procedures, we generated indications that

  8. Atmospheric Profile Imprint in Firewall Ablation Coefficient

    Science.gov (United States)

    Ceplecha, Z.; Pecina, P.

    1984-01-01

    A general formula which expresses the distance along the meteoric fireball trajectory 1 as a function of t is discussed. Differential equations which include the motion and ablation of a single nonfragmenting meteor body are presented. The importance of the atmospheric density profile in the meteor formula is emphasized.

  9. Ablation of liver metastases by radiofrequency

    International Nuclear Information System (INIS)

    Baere, T. de

    2012-01-01

    Radiofrequency is a thermal ablative technique that is most often used percuteanously under image guidance. Thermal damage is obtained through frictional heating of a high frequency current. The maximal volume of destruction obtained in one radiofrequency delivery is around 4 cm and consequently, best indication for treatment are tumours below 3 cm. When compared, radiofrequency and surgical removal for tumours below 25 mm in diameter demonstrated a rate of incomplete resection/ablation of 6% and 7.3% respectively. Median survival after the first radiofrequency of a liver metastasis of CRC is reported to be 24 to 52 months with a 5 years overall survival of 18 to 44%. The median overall survival increases from 22 to 48 months depending on the use of radiofrequency ablation as rescue treatment after failure of others, or as a first line treatment. For patients with a single tumour, less than 4 cm, the survival rates at 1, 3, and 5 years are respectively 97%, 84% and 40%, with a median survival of 50 months. Follow-up imaging requires to use contrast-enhanced CT or MRI, looking for local recurrences evidenced by local foci of enhancement at the periphery of the ablation zone. (author)

  10. Pulsed Radiofrequency Ablation for Treating Sural Neuralgia.

    Science.gov (United States)

    Abd-Elsayed, Alaa; Jackson, Markus; Plovanich, Elizabeth

    2018-01-01

    Sural neuralgia is persistent pain in the distribution of the sural nerve that provides sensation to the lateral posterior corner of the leg, lateral foot, and fifth toe. Sural neuralgia is a rare condition but can be challenging to treat and can cause significant limitation. We present 2 cases of sural neuralgia resistant to conservative management that were effectively treated by pulsed radiofrequency ablation. A 65-year-old female developed sural neuralgia after a foot surgery and failed conservative management. She had successful sural nerve blocks, and pulsed radiofrequency ablation led to an 80% improvement in her pain. A 33-year-old female presented with sural neuralgia secondary to two falls. The patient had tried several conservative modalities with no success. We performed diagnostic blocks and pulsed radiofrequency ablation, and the patient reported 80% improvement in her pain. Pulsed radiofrequency ablation may be a safe and effective treatment for patients with sural neuralgia that does not respond to conservative therapy. However, studies are needed to elucidate its effectiveness and safety profile.

  11. Organized Atrial Tachycardias after Atrial Fibrillation Ablation

    Science.gov (United States)

    Castrejón-Castrejón, Sergio; Ortega, Marta; Pérez-Silva, Armando; Doiny, David; Estrada, Alejandro; Filgueiras, David; López-Sendón, José L.; Merino, José L.

    2011-01-01

    The efficacy of catheter-based ablation techniques to treat atrial fibrillation is limited not only by recurrences of this arrhythmia but also, and not less importantly, by new-onset organized atrial tachycardias. The incidence of such tachycardias depends on the type and duration of the baseline atrial fibrillation and specially on the ablation technique which was used during the index procedure. It has been repeatedly reported that the more extensive the left atrial surface ablated, the higher the incidence of organized atrial tachycardias. The exact origin of the pathologic substrate of these trachycardias is not fully understood and may result from the interaction between preexistent regions with abnormal electrical properties and the new ones resultant from radiofrequency delivery. From a clinical point of view these atrial tachycardias tend to remit after a variable time but in some cases are responsible for significant symptoms. A precise knowledge of the most frequent types of these arrhythmias, of their mechanisms and components is necessary for a thorough electrophysiologic characterization if a new ablation procedure is required. PMID:21941669

  12. Microwave ablation devices for interventional oncology.

    Science.gov (United States)

    Ward, Robert C; Healey, Terrance T; Dupuy, Damian E

    2013-03-01

    Microwave ablation is one of the several options in the ablation armamentarium for the treatment of malignancy, offering several potential benefits when compared with other ablation, radiation, surgical and medical treatment modalities. The basic microwave system consists of the generator, power distribution system and antennas. Often under image (computed tomography or ultrasound) guidance, a needle-like antenna is inserted percutaneously into the tumor, where local microwave electromagnetic radiation is emitted from the probe's active tip, producing frictional tissue heating, capable of causing cell death by coagulation necrosis. Half of the microwave ablation systems use a 915 MHz generator and the other half use a 2450 MHz generator. To date, there are no completed clinical trials comparing microwave devices head-to-head. Prospective comparisons of microwave technology with other treatment alternatives, as well as head-to-head comparison with each microwave device, is needed if this promising field will garner more widespread support and use in the oncology community.

  13. Plasma dynamics from laser ablated solid lithium

    Indian Academy of Sciences (India)

    b; 52.25.-b; 52.70.-m. 1. Introduction. Pulsed laser ablation of a solid sample generates a dense plasma emission in the shape of ... The multichannel analyser plate of the ICCD was gated for as less as 4 ns using ... to explain the atomic collision processes [4]. .... Within duration of laser pulse, there occurs laser-solid interac-.

  14. Bending diamonds by femtosecond laser ablation

    DEFF Research Database (Denmark)

    Balling, Peter; Esberg, Jakob; Kirsebom, Kim

    2009-01-01

    We present a new method based on femtosecond laser ablation for the fabrication of statically bent diamond crystals. Using this method, curvature radii of 1 m can easily be achieved, and the curvature obtained is very uniform. Since diamond is extremely tolerant to high radiation doses, partly due...

  15. Femtosecond laser ablation of bovine cortical bone

    Science.gov (United States)

    Cangueiro, Liliana T.; Vilar, Rui; Botelho do Rego, Ana M.; Muralha, Vania S. F.

    2012-12-01

    We study the surface topographical, structural, and compositional modifications induced in bovine cortical bone by femtosecond laser ablation. The tests are performed in air, with a Yb:KYW chirped-pulse-regenerative amplification laser system (500 fs, 1030 nm) at fluences ranging from 0.55 to 2.24 J/cm2. The ablation process is monitored by acoustic emission measurements. The topography of the laser-treated surfaces is studied by scanning electron microscopy, and their constitution is characterized by glancing incidence x-ray diffraction, x-ray photoelectron spectroscopy, Fourier transform infrared spectroscopy, and micro-Raman spectroscopy. The results show that femtosecond laser ablation allows removing bone without melting, carbonization, or cracking. The structure and composition of the remaining tissue are essentially preserved, the only constitutional changes observed being a reduction of the organic material content and a partial recrystallization of hydroxyapatite in the most superficial region of samples. The results suggest that, within this fluence range, ablation occurs by a combination of thermal and electrostatic mechanisms, with the first type of mechanism predominating at lower fluences. The associated thermal effects explain the constitutional changes observed. We show that femtosecond lasers are a promising tool for delicate orthopaedic surgeries, where small amounts of bone must be cut with negligible damage, thus minimizing surgical trauma.

  16. Plume collimation for laser ablation electrospray ionization mass spectrometry

    Science.gov (United States)

    Vertes, Akos; Stolee, Jessica A.

    2014-09-09

    In various embodiments, a device may generally comprise a capillary having a first end and a second end; a laser to emit energy at a sample in the capillary to ablate the sample and generate an ablation plume in the capillary; an electrospray apparatus to generate an electrospray plume to intercept the ablation plume to produce ions; and a mass spectrometer having an ion transfer inlet to capture the ions. The ablation plume may comprise a collimated ablation plume. The device may comprise a flow cytometer. Methods of making and using the same are also described.

  17. Laser ablation comparison by picosecond pulses train and nanosecond pulse

    Science.gov (United States)

    Lednev, V. N.; Filippov, M. N.; Bunkin, A. F.; Pershin, S. M.

    2015-12-01

    A comparison of laser ablation by a train of picosecond pulses and nanosecond pulses revealed a difference in laser craters, ablation thresholds, plasma sizes and spectral line intensities. Laser ablation with a train of picosecond pulses resulted in improved crater quality while ablated mass decreased up to 30%. A reduction in laser plasma dimensions for picosecond train ablation was observed while the intensity of atomic/ionic lines in the plasma spectra was greater by a factor of 2-4 indicating an improved excitation and atomization in the plasma.

  18. Burn, freeze, or photo-ablate?: comparative symptom profile in Barrett's dysplasia patients undergoing endoscopic ablation

    Science.gov (United States)

    Gill, Kanwar Rupinder S.; Gross, Seth A.; Greenwald, Bruce D.; Hemminger, Lois L.; Wolfsen, Herbert C.

    2009-06-01

    Background: There are few data available comparing endoscopic ablation methods for Barrett's esophagus with high-grade dysplasia (BE-HGD). Objective: To determine differences in symptoms and complications associated with endoscopic ablation. Design: Prospective observational study. Setting: Two tertiary care centers in USA. Patients: Consecutive patients with BE-HGD Interventions: In this pilot study, symptoms profile data were collected for BE-HGD patients among 3 endoscopic ablation methods: porfimer sodium photodynamic therapy, radiofrequency ablation and low-pressure liquid nitrogen spray cryotherapy. Main Outcome Measurements: Symptom profiles and complications from the procedures were assessed 1-8 weeks after treatment. Results: Ten BE-HGD patients were treated with each ablation modality (30 patients total; 25 men, median age: 69 years (range 53-81). All procedures were performed in the clinic setting and none required subsequent hospitalization. The most common symptoms among all therapies were chest pain, dysphagia and odynophagia. More patients (n=8) in the porfimer sodium photodynamic therapy group reported weight loss compared to radio-frequency ablactation (n=2) and cryotherapy (n=0). Four patients in the porfimer sodium photodynamic therapy group developed phototoxicity requiring medical treatment. Strictures, each requiring a single dilation, were found in radiofrequency ablactation (n=1) and porfimer sodium photodynamic therapy (n=2) patients. Limitations: Small sample size, non-randomized study. Conclusions: These three endoscopic therapies are associated with different types and severity of post-ablation symptoms and complications.

  19. Effects of pressure rise on cw laser ablation of tissue

    Science.gov (United States)

    LeCarpentier, Gerald L.; Motamedi, Massoud; Welch, Ashley J.

    1991-06-01

    The objectives of this research were to identify mechanisms responsible for the initiation of continuous wave (cw) laser ablation of tissue and investigate the role of pressure in the ablation process. Porcine aorta samples were irradiated in a chamber pressurized from 1 X 10-4 to 12 atmospheres absolute pressure. Acrylic and Zn-Se windows in the experimental pressure chamber allowed video and infrared cameras to simultaneously record mechanical and thermal events associated with cw argon laser ablation of these samples. Video and thermal images of tissue slabs documented the explosive nature of cw laser ablation of soft biological media and revealed similar ablation threshold temperatures and ablation onset times under different environmental pressures; however, more violent initiation explosions with decreasing environmental pressures were observed. These results suggest that ablation initiates with thermal alterations in the mechanical strength of the tissue and proceeds with an explosion induced by the presence superheated liquid within the tissue.

  20. Thermal Ablation of T1c Renal Cell Carcinoma: A Comparative Assessment of Technical Performance, Procedural Outcome, and Safety of Microwave Ablation, Radiofrequency Ablation, and Cryoablation.

    Science.gov (United States)

    Zhou, Wenhui; Arellano, Ronald S

    2018-04-06

    To evaluate perioperative outcomes of thermal ablation with microwave (MW), radiofrequency (RF), and cryoablation for stage T1c renal cell carcinoma (RCC). A retrospective analysis of 384 patients (mean age, 71 y; range, 22-88 y) was performed between October 2006 and October 2016. Mean radius, exophytic/endophytic, nearness to collecting system or sinus, anterior/posterior, and location relative to polar lines; preoperative aspects and dimensions used for anatomic classification; and centrality index scores were 6.3, 7.9, and 2.7, respectively. Assessment of pre- and postablation serum blood urea nitrogen, creatinine, and estimated glomerular filtration rate was performed to assess functional outcomes. Linear regression analyses were performed to compare sedation medication dosages among the three treatment cohorts. Univariable and multivariable logistic regression analyses were performed to compare rates of residual disease and complications among treatment modalities. A total of 437 clinical stage T1N0M0 biopsy-proven RCCs measuring 1.2-6.9 cm were treated with computed tomography (CT)-guided MW ablation (n = 44; 10%), RF ablation (n = 347; 79%), or cryoablation (n = 46; 11%). There were no significant differences in patient demographic or tumor characteristics among cohorts. Complication rates and immediate renal function changes were similar among the three ablation modalities (P = .46 and P = .08, respectively). MW ablation was associated with significantly decreased ablation time (P < .05), procedural time (P < .05), and dosage of sedative medication (P < .05) compared with RF ablation and cryoablation. CT-guided percutaneous MW ablation is comparable to RF ablation or cryoablation for the treatment of stage T1N0M0 RCC with regard to treatment response and is associated with shorter treatment times and less sedation than RF ablation or cryoablation. In addition, the safety profile of CT-guided MW ablation is noninferior to those of RF ablation or

  1. Tetraspanin CD63 Bridges Autophagic and Endosomal Processes To Regulate Exosomal Secretion and Intracellular Signaling of Epstein-Barr Virus LMP1

    Science.gov (United States)

    Hurwitz, Stephanie N; Cheerathodi, Mujeeb R; Nkosi, Dingani; York, Sara B; Meckes, David G

    2018-03-01

    The tetraspanin protein CD63 has been recently described as a key factor in extracellular vesicle (EV) production and endosomal cargo sorting. In the context of Epstein-Barr virus (EBV) infection, CD63 is required for the efficient packaging of the major viral oncoprotein latent membrane protein 1 (LMP1) into exosomes and other EV populations and acts as a negative regulator of LMP1 intracellular signaling. Accumulating evidence has also pointed to intersections of the endosomal and autophagy pathways in maintaining cellular secretory processes and as sites for viral assembly and replication. Indeed, LMP1 can activate the mammalian target of rapamycin (mTOR) pathway to suppress host cell autophagy and facilitate cell growth and proliferation. Despite the growing recognition of cross talk between endosomes and autophagosomes and its relevance to viral infection, little is understood about the molecular mechanisms governing endosomal and autophagy convergence. Here, we demonstrate that CD63-dependent vesicle protein secretion directly opposes intracellular signaling activation downstream of LMP1, including mTOR-associated proteins. Conversely, disruption of normal autolysosomal processes increases LMP1 secretion and dampens signal transduction by the viral protein. Increases in mTOR activation following CD63 knockout are coincident with the development of serum-dependent autophagic vacuoles that are acidified in the presence of high LMP1 levels. Altogether, these findings suggest a key role of CD63 in regulating the interactions between endosomal and autophagy processes and limiting cellular signaling activity in both noninfected and virally infected cells. IMPORTANCE The close connection between extracellular vesicles and viruses is becoming rapidly and more widely appreciated. EBV, a human gamma herpesvirus that contributes to the progression of a multitude of lymphomas and carcinomas in immunocompromised or genetically susceptible populations, packages its major

  2. Study on the lithium compound clusters using laser ablation

    International Nuclear Information System (INIS)

    Yokoyama, Keiichi

    2001-01-01

    Lithium-rich nonstoichiometric binary clusters including hyperlithiated molecules were found to be produced by a nanosecond laser ablation of lithium metal or compound target. Structural information on Li 3 O was obtained for the first time from experiments by measuring and analyzing photoionization efficiency curves of mass-selected ions. For example, the structure of Li 3 O was concluded to have both D 3h and C 2v symmetry. In other words, the vibrational wavefunction even at the ground state spreads over the C 2v and D 3h minima, which has been predicted as the global minimum in the latest theoretical calculations. Also, this is the first experimental evidence for electronomers'. (author)

  3. Five-year survival in 309 patients with colorectal liver metastases treated with radiofrequency ablation

    International Nuclear Information System (INIS)

    Gillams, A.R.; Lees, W.R.

    2009-01-01

    There is little published long-term survival data for patients with colorectal liver metastases treated with radiofrequency ablation (RFA). We present a multivariate analysis of 5-year survival in 309 patients (198 male, aged 64 (24-92)) treated at 617 sessions. Our standard protocol used internally cooled electrodes introduced percutaneously under combined US and CT guidance/monitoring. The number and size of liver metastases, the presence and location of extrahepatic disease, primary resection, clinical, chemotherapy and follow-up data were recorded. Data analysis was performed using SPSS v.10. On multivariate analysis, significant survival factors were the presence of extrahepatic disease (p < 0.001) and liver tumour volume (p = 0.001). For 123 patients with five or less metastases of 5 cm or less maximum diameter and no extrahepatic disease median survival was 46 and 36 months from liver metastasis diagnosis and ablation, respectively; corresponding 3- and 5-year survival rates were 63%, 34% and 49%, 24%. Sixty-nine patients had three or less tumours of below 3.5 cm in diameter and their 5-year survival from ablation was 33%. There were 23/617(3.7%) local complications requiring intervention. Five-year survival of 24-33% post ablation in selected patients is superior to any published chemotherapy data and approaches the results of liver resection. (orig.)

  4. Pulmonary hemorrhage complicating radiofrequency ablation, from mild hemoptysis to life-threatening pattern

    Energy Technology Data Exchange (ETDEWEB)

    Nour-Eldin, Nour-Eldin A.; Naguib, Nagy N.N.; Mack, Martin; Abskharon, John E.; Vogl, Thomas J. [Johann Wolfgang Goethe-University Hospital, Institute for Diagnostic and Interventional Radiology, Frankfurt am Main (Germany)

    2011-01-15

    To assess risk factors and the extent of pulmonary hemorrhage complicating radiofrequency ablation (RFA) of pulmonary neoplasms. This retrospective study involved 248 ablation sessions for lung tumors (20 primary lesions and 228 metastatic lesions) in 164 patients (mean age 59.7 years, SD: 10.2). Both unipolar and bipolar radiofrequency systems were used under CT fluoroscopic guidance. Extent and underlying factors associated with development of pulmonary hemorrhage were analyzed. Incidence of intra-parenchymal pulmonary hemorrhage, pleural effusion, and hemoptysis were 17.7% (44/248 sessions), 4% (8/248 sessions), and 16.1% (40/248 sessions), respectively. Death because of massive bleeding occurred in one session (0.4%). Significant risk factors associated with intra-parenchymal hemorrhage included: lesions of <1.5 cm diameter (P = 0.007); basal and middle lung zone lesions (P = 0.026); increased needle track distance traversing the lung parenchyma >2.5 cm (P = 0.0017); traversing pulmonary vessels in the track of ablation (P < 0.001); and the use of multi-tined electrodes (P = 0.004). Concomitant incidence of pulmonary hemorrhage and pneumothorax was 29.2% (14/48 sessions). While typically safe, RFA of pulmonary neoplasms can result in pulmonary hemorrhage ranging from mild to life-threatening. Management of this complication is mainly preventive through adequate patient selection for ablation therapy and exclusion of technically avoidable risk factors. (orig.)

  5. Percutaneous ultrasound-guided radiofrequency ablation for kidney tumors in patients with surgical risk

    International Nuclear Information System (INIS)

    Salagierski, Marek; Salagierski, Maciej; Sosnowski, Marek; Salagierska-Barwinska, Anna

    2006-01-01

    The aim of this study was to describe our experience with percutaneous ultrasound-guided radiofrequency ablation of kidney tumors. From July 2002 to August 2005, 45 radiofrequency ablations (RFA) in 42 selected patients with kidney tumor were performed. The patients had either contraindications to surgery procedures or had a solitary kidney. The average tumor size was 37.5 mm (range, 18-59 mm) with the mean age of 68 years (range, 28-83 years). RFA were performed based on radiographic findings. Needle biopsy was made only twice. Monopolar Cool-tip Tyco or bipolar Celon Olympus radiofrequency devices were used. The procedure was performed under conscious sedation with local anesthesia. Treatment efficacy was assessed by computed tomography and by Doppler ultrasound. The absence of contrast enhancement on computed tomography was considered to be a successful treatment. The average follow up was 14 months (range, 3-36 months). In 42 tumors (93%), total absence of contrast enhancement was obtained after the initial RFA and in three tumors (7%) after the second ablation session. There were no complications following 41 procedures, including all ablations in small (<35 mm) renal masses. In four procedures, minor complications were observed. All patients are alive. There has been no need for chronic hemodialysis and, until now, we have not observed any local recurrences with the exception of one metastasis to an ipsilateral adrenal gland. RFA of kidney tumors is a promising alternative treatment which could be considered for patients who are not suitable for surgery. (author)

  6. Anorexia and impaired glucose metabolism in mice with hypothalamic ablation of Glut4 neurons.

    Science.gov (United States)

    Ren, Hongxia; Lu, Taylor Y; McGraw, Timothy E; Accili, Domenico

    2015-02-01

    The central nervous system (CNS) uses glucose independent of insulin. Nonetheless, insulin receptors and insulin-responsive glucose transporters (Glut4) often colocalize in neurons (Glut4 neurons) in anatomically and functionally distinct areas of the CNS. The apparent heterogeneity of Glut4 neurons has thus far thwarted attempts to understand their function. To answer this question, we used Cre-dependent, diphtheria toxin-mediated cell ablation to selectively remove basal hypothalamic Glut4 neurons and investigate the resulting phenotypes. After Glut4 neuron ablation, mice demonstrate altered hormone and nutrient signaling in the CNS. Accordingly, they exhibit negative energy balance phenotype characterized by reduced food intake and increased energy expenditure, without locomotor deficits or gross neuronal abnormalities. Glut4 neuron ablation affects orexigenic melanin-concentrating hormone neurons but has limited effect on neuropeptide Y/agouti-related protein and proopiomelanocortin neurons. The food intake phenotype can be partially normalized by GABA administration, suggesting that it arises from defective GABAergic transmission. Glut4 neuron-ablated mice show peripheral metabolic defects, including fasting hyperglycemia and glucose intolerance, decreased insulin levels, and elevated hepatic gluconeogenic genes. We conclude that Glut4 neurons integrate hormonal and nutritional cues and mediate CNS actions of insulin on energy balance and peripheral metabolism. © 2015 by the American Diabetes Association. Readers may use this article as long as the work is properly cited, the use is educational and not for profit, and the work is not altered.

  7. The effect of ethanol infusion on the size of the ablated lesion in radiofrequency thermal ablation: A pilot study

    International Nuclear Information System (INIS)

    Kim, Young Sun; Rhim, Hyun Chul; Koh, Byung Hee; Cho, On Koo; Seo, Heung Suk; Kim, Yong Soo; Joo, Kyoung Bin

    2001-01-01

    To assess the effect of ethanol infusion on the size of ablated lesion during radiofrequency (RF) thermal ablation. We performed an ex vivo experimental study using a total of 15 pig livers. Three groups were designed: 1)normal control (n=10), 2) saline infusion (n=10) 3) ethanol infusion (n=10). Two radiofrequency ablations were done using a 50 watt RF generator and a 15 guage expandable elections with four prongs in each liver. During ablation for 8 minutes, continuous infusion of fluid at a rate of 0.5 ml/min through the side arm of electrode was performed. We checked the frequency of the 'impeded-out' phenomenon due to abrupt increase of impedance during ablation. Size of ablated lesion was measured according to length, width, height, and subsequently volume after the ablations. The sizes of the ablated lesions were compared between the three groups. 'Impeded-out' phenomenon during ablation was noted 4 times in control group, although that never happened in saline or ethanol infusion groups. There were significant differences in the volumes of ablated lesions between control group (10.62 ± 1.45 cm 3 ) and saline infusion group (15.33 ± 2.47 cm 3 ), and saline infusion group and ethanol infusion group (18.78 ± 3.58 cm 3 ) (p<0.05). Fluid infusion during radiofrequency thermal ablation decrease a chance of charming and increase the volume of the ablated lesion. Ethanol infusion during ablation may induce larger volume of ablated lesion than saline infusion.

  8. Value of local electrogram characteristics predicting successful catheter ablation of left-versus right-sided accessory atrioventricular pathways by radiofrequency current.

    Science.gov (United States)

    Lin, J L; Schie, J T; Tseng, C D; Chen, W J; Cheng, T F; Tsou, S S; Chen, J J; Tseng, Y Z; Lien, W P

    1995-01-01

    Despite similar guidance by local electrogram criteria, catheter ablation of right-sided accessory atrioventricular (AV) pathways by radiofrequency current has been less effective than that of left-sided ones. In order to elucidate the possible diversities in local electrosignal criteria, we systematically analyzed the morphological and timing characteristics of 215 bipolar local electrograms from catheter ablation sites of 65 left-sided accessory AV pathways and of 356 from those of 37 right-sided ones in 92 consecutive patients with Wolff-Parkinson-White syndrome or AV reentrant tachycardia incorporating concealed accessory AV pathways. After stepwise multivariate analysis, we selected the presence of a possible accessory pathway potential, local ventricular activation preceding QRS complex for 20 ms or more during ventricular insertion mapping, and the local retrograde ventriculoatrial (VA) continuity, local retrograde VA interval right-sided targets only) during atrial insertion mapping, as independent local electrogram predictors for successful ablation of left- and right-sided accessory AV pathways. Combination of all local electrogram predictors could have moderate chance of success (80 and 51%) for the ventricular and atrial insertion ablation of left-sided accessory AV pathways, but only low probability of success (40% in ventricular insertion ablation) or very low sensitivity (12.5% in atrial insertion ablation) for right-sided ones. In conclusion, with the present approach, successful catheter ablation of right-sided accessory AV pathways, compared to left-sided ones, still necessitate a breakthrough in the precision mapping and the efficiency of energy delivery.

  9. Automated planning of ablation targets in atrial fibrillation treatment

    Science.gov (United States)

    Keustermans, Johannes; De Buck, Stijn; Heidbüchel, Hein; Suetens, Paul

    2011-03-01

    Catheter based radio-frequency ablation is used as an invasive treatment of atrial fibrillation. This procedure is often guided by the use of 3D anatomical models obtained from CT, MRI or rotational angiography. During the intervention the operator accurately guides the catheter to prespecified target ablation lines. The planning stage, however, can be time consuming and operator dependent which is suboptimal both from a cost and health perspective. Therefore, we present a novel statistical model-based algorithm for locating ablation targets from 3D rotational angiography images. Based on a training data set of 20 patients, consisting of 3D rotational angiography images with 30 manually indicated ablation points, a statistical local appearance and shape model is built. The local appearance model is based on local image descriptors to capture the intensity patterns around each ablation point. The local shape model is constructed by embedding the ablation points in an undirected graph and imposing that each ablation point only interacts with its neighbors. Identifying the ablation points on a new 3D rotational angiography image is performed by proposing a set of possible candidate locations for each ablation point, as such, converting the problem into a labeling problem. The algorithm is validated using a leave-one-out-approach on the training data set, by computing the distance between the ablation lines obtained by the algorithm and the manually identified ablation points. The distance error is equal to 3.8+/-2.9 mm. As ablation lesion size is around 5-7 mm, automated planning of ablation targets by the presented approach is sufficiently accurate.

  10. Percutaneous thermal ablation of renal neoplasms; Perkutane Thermoablation von Nierentumoren

    Energy Technology Data Exchange (ETDEWEB)

    Tacke, J. [Inst. fuer Diagnostische und Interventionelle Radiologie/Neuroradiologie, Klinikum Passau (Germany); Mahnken, A.H.; Guenther, R.W. [Klinik fuer Radiologische Diagnostik, Universitaetsklinikum Aachen (Germany)

    2005-12-15

    Due to modern examination techniques such as multidetector computed tomography and high-field magnetic resonance imaging, the detection rate of renal neoplasms is continually increasing. Even though tumors exceeding 4 cm in diameter rarely metastasize, all renal lesions that are possible neoplasms should be treated. Traditional treatment techniques include radical nephrectomy or nephron-sparing resection, which are increasingly performed laparoscopically. Modern thermal ablation techniques such as hyperthermal techniques like radiofrequency ablation RFA, laser induced thermal ablation LITT, focused ultrasound FUS and microwave therapy MW, as well as hypothermal techniques (cryotherapy) may be a useful treatment option for patients who are unfit for or refuse surgical resection. Cryotherapy is the oldest and best known thermal ablation technique and can be performed laparoscopically or percutaneously. Since subzero temperatures have no antistyptic effect, additional maneuvers must be performed to control bleeding. Percutaneous cryotherapy of renal tumors is a new and interesting method, but experience with it is still limited. Radiofrequency ablation is the most frequently used method. Modern probe design allows volumes between 2 and 5 cm in diameter to be ablated. Due to hyperthermal tract ablation, the procedure is deemed to be safe and has a low complication rate. Although there are no randomized comparative studies to open resection, the preliminary results for renal RFA are promising and show RFA to be superior to other thermal ablation techniques. Clinical success rates are over 90% for both, cryo- and radiofrequency ablation. Whereas laser induced thermal therapy is established in hepatic ablation, experience is minimal with respect to renal application. For lesions of more than 2 cm in diameter, additional cooling catheters are required. MR thermometry offers temperature control during ablation. Microwave ablation is characterized by small ablation volumes

  11. Device-based approaches for renal nerve ablation for hypertension and beyond

    OpenAIRE

    Alicia Ann Thorp; Markus Peter Schlaich; Markus Peter Schlaich

    2015-01-01

    Animal and human studies have demonstrated that chronic activation of renal sympathetic nerves is critical in the pathogenesis and perpetuation of treatment-resistant hypertension. Bilateral renal denervation has emerged as a safe and effective, non-pharmacological treatment for resistant hypertension that involves the selective ablation of efferent and afferent renal nerves to lower blood pressure. However, the most recent and largest randomized controlled trial failed to confirm the primacy...

  12. Anorexia and Impaired Glucose Metabolism in Mice With Hypothalamic Ablation of Glut4 Neurons

    OpenAIRE

    Ren, Hongxia; Lu, Taylor Y.; McGraw, Timothy E.; Accili, Domenico

    2014-01-01

    The central nervous system (CNS) uses glucose independent of insulin. Nonetheless, insulin receptors and insulin-responsive glucose transporters (Glut4) often colocalize in neurons (Glut4 neurons) in anatomically and functionally distinct areas of the CNS. The apparent heterogeneity of Glut4 neurons has thus far thwarted attempts to understand their function. To answer this question, we used Cre-dependent, diphtheria toxin?mediated cell ablation to selectively remove basal hypothalamic Glut4 ...

  13. Transmission Geometry Laser Ablation into a Non-Contact Liquid Vortex Capture Probe for Mass Spectrometry Imaging

    Energy Technology Data Exchange (ETDEWEB)

    Ovchinnikova, Olga S [ORNL; Bhandari, Deepak [ORNL; Lorenz, Matthias [ORNL; Van Berkel, Gary J [ORNL

    2014-01-01

    chemical image. This imaging resolution was 20 times better than the previous best reported results with laser ablation/liquid sample capture mass spectrometry imaging. Using thin sections of brain tissue the chemical image of a selected lipid was obtained with an estimated imaging resolution of about 50 um. Conclusions: A vertically aligned, transmission geometry laser ablation liquid vortex capture probe, electrospray ionization mass spectrometry system provides an effective means for spatially resolved spot sampling and imaging with mass spectrometry.

  14. Impact of chronic lymphocytic thyroiditis co-existing with differentiated thyroid cancer on the effectiveness of remnants ablation

    International Nuclear Information System (INIS)

    Boughattas, S.; Chatti, K.; Trimeche, M.; Mokni, M.

    2004-01-01

    Full text: Some stages of chronic lymphocytic thyroiditis (CLT) are functionally characterized by an organification defect with large intra thyroid inorganic iodide pool, which can be discharged during perchlorate test. Fluorescent scan study indicates that most patients with CLT have decreased stable iodine store in the thyroid gland. The aim of our study was to investigate the possible consequences of these organification abnormalities during remnants ablation in patients with coexisting differentiated thyroid cancer (DTC) and chronic lymphocytic thyroiditis. We reviewed our series of patients of DTC being followed at the department of nuclear medicine of the university hospital Sahloul. Among the 350 patients with DTC, 30 (8.5%) had histologically proved chronic lymphocytic thyroiditis, with infiltration of the non-tumoral thyroid tissue. A second group of 60 patients (without evidence of lymphocytic infiltration) was selected randomly and used as control. The median of follow-up for these two groups was 4 years. All patients had undergone total thyroidectomy followed by scintigraphy 4-6 weeks later. In patients with thyroid remnants, standard ablative dose of 3.7 GBq of I-131 (100 mCi) was administrated with 6 months duration between all therapies, until the negativity of thyroid bed activity on follow up survey scan performed 48 to 72 hours after administering 2 to 3 mCi of I-131. Thyroglobulin (Tg) serum level was not considered as a criterion of ablation, because of the frequency of anti-thyroid antibodies in CLT. In the group with CLT, 3 patients had negative postoperative neck scintigraphy. Complete ablation was achieved with a single standard dose in 14, two standard doses in 5, and more than 200 mCi in two patients (300 in one and 400 in two). In five patients, ablation is not yet achieved. In the control group, ablation was obtained with 100 mCi in 43 patients, 200 mCi in 9, and 300 mCi in 3. In five patients ablation has not been achieved. Considering

  15. 3D Multifunctional Ablative Thermal Protection System

    Science.gov (United States)

    Feldman, Jay; Venkatapathy, Ethiraj; Wilkinson, Curt; Mercer, Ken

    2015-01-01

    NASA is developing the Orion spacecraft to carry astronauts farther into the solar system than ever before, with human exploration of Mars as its ultimate goal. One of the technologies required to enable this advanced, Apollo-shaped capsule is a 3-dimensional quartz fiber composite for the vehicle's compression pad. During its mission, the compression pad serves first as a structural component and later as an ablative heat shield, partially consumed on Earth re-entry. This presentation will summarize the development of a new 3D quartz cyanate ester composite material, 3-Dimensional Multifunctional Ablative Thermal Protection System (3D-MAT), designed to meet the mission requirements for the Orion compression pad. Manufacturing development, aerothermal (arc-jet) testing, structural performance, and the overall status of material development for the 2018 EM-1 flight test will be discussed.

  16. Does Left Atrial Volume and Pulmonary Venous Anatomy Predict the Outcome of Catheter Ablation of Atrial Fibrillation ?

    NARCIS (Netherlands)

    Hof, Irene; Chilukuri, Karuna; Arbab-Zadeh, Armin; Scherr, Daniel; Dalal, Darshan; Nazarian, Saman; Henrikson, Charles; Spragg, David; Berger, Ronald; Marine, Joseph; Calkins, Hugh

    Introduction: Preprocedural factors may be helpful in selecting patients with atrial fibrillation (AF) for treatment with catheter ablation and in making an assumption regarding their prognosis. The aims of this study were to investigate whether left atrial (LA) volume and pulmonary venous (PV)

  17. Design calculations for NIF convergent ablator experiments

    Directory of Open Access Journals (Sweden)

    Olson R.E.

    2013-11-01

    Full Text Available The NIF convergent ablation tuning effort is underway. In the early experiments, we have discovered that the design code simulations over-predict the capsule implosion velocity and shock flash ρr, but under-predict the hohlraum x-ray flux measurements. The apparent inconsistency between the x-ray flux and radiography data implies that there are important unexplained aspects of the hohlraum and/or capsule behavior.

  18. Radiofrequency ablation of two femoral head chondroblastomas

    Energy Technology Data Exchange (ETDEWEB)

    Petsas, Theodore [Department of Radiology, University of Patras (Greece); Megas, Panagiotis [Department of Orthopaedic Surgery, University of Patras (Greece)]. E-mail: panmegas@med.upatras.gr; Papathanassiou, Zafiria [Department of Radiology, University of Patras (Greece)

    2007-07-15

    Chondroblastoma is a rare benign cartilaginous bone tumor. Surgical resection is the treatment of choice for pain relief and prevention of further growth. Open surgical techniques are associated with complications, particularly when the tumors are located in deep anatomical sites. The authors performed RF ablation in two cases of subarticular femoral head chondroblastomas and emphasize its positive impact. The clinical course, the radiological findings and the post treatment results are discussed.

  19. A Review of Laser Ablation Propulsion

    International Nuclear Information System (INIS)

    Phipps, Claude; Bohn, Willy; Lippert, Thomas; Sasoh, Akihiro; Schall, Wolfgang; Sinko, John

    2010-01-01

    Laser Ablation Propulsion is a broad field with a wide range of applications. We review the 30-year history of laser ablation propulsion from the transition from earlier pure photon propulsion concepts of Oberth and Saenger through Kantrowitz's original laser ablation propulsion idea to the development of air-breathing 'Lightcraft' and advanced spacecraft propulsion engines. The polymers POM and GAP have played an important role in experiments and liquid ablation fuels show great promise. Some applications use a laser system which is distant from the propelled object, for example, on another spacecraft, the Earth or a planet. Others use a laser that is part of the spacecraft propulsion system on the spacecraft. Propulsion is produced when an intense laser beam strikes a condensed matter surface and produces a vapor or plasma jet. The advantages of this idea are that exhaust velocity of the propulsion engine covers a broader range than is available from chemistry, that it can be varied to meet the instantaneous demands of the particular mission, and that practical realizations give lower mass and greater simplicity for a payload delivery system. We review the underlying theory, buttressed by extensive experimental data. The primary problem in laser space propulsion theory has been the absence of a way to predict thrust and specific impulse over the transition from the vapor to the plasma regimes. We briefly discuss a method for combining two new vapor regime treatments with plasma regime theory, giving a smooth transition from one regime to the other. We conclude with a section on future directions.

  20. Rabies virus co-localizes with early (Rab5) and late (Rab7) endosomal proteins in neuronal and SH-SY5Y cells.

    Science.gov (United States)

    Ahmad, Waqas; Li, Yingying; Guo, Yidi; Wang, Xinyu; Duan, Ming; Guan, Zhenhong; Liu, Zengshan; Zhang, Maolin

    2017-06-01

    Rabies virus (RABV) is a highly neurotropic virus that follows clathrin-mediated endocytosis and pH-dependent pathway for trafficking and invasion into endothelial cells. Early (Rab5, EEA1) and late (Rab7, LAMP1) endosomal proteins play critical roles in endosomal sorting, maturity and targeting various molecular cargoes, but their precise functions in the early stage of RABV neuronal infection remain elusive. In this study, the relationship between enigmatic entry of RABV with these endosomal proteins into neuronal and SH-SY5Y cells was investigated. Immunofluorescence, TCID 50 titers, electron microscopy and western blotting were carried out to determine the molecular interaction of the nucleoprotein (N) of RABV with early or late endosomal proteins in these cell lines. The expression of N was also determined by down-regulating Rab5 and Rab7 in both cell lines through RNA interference. The results were indicative that N proficiently colocalized with Rab5/EEA1 and Rab7/LAMP1 in both cell lines at 24 and 48 h post-infection, while N titers significantly decreased in early infection of RABV. Down-regulation of Rab5 and Rab7 did not inhibit N expression, but it prevented productive infection via blocking the normal trafficking of RABV in a low pH environment. Ultrathin sections of cells studied by electron microscope also verified the close association of RABV with Rab5 and Rab7 in neurons. From the data it was concluded that primary entry of RABV strongly correlates with the kinetics of Rab-proteins present on early and late vesicles, which provides helpful clues to explain the early events of RABV in nerve cells.

  1. Strategies for the Activation and Release of the Membranolytic Peptide Melittin from Liposomes Using Endosomal pH as a Trigger.

    Science.gov (United States)

    Oude Blenke, E; Sleszynska, M; Evers, M J W; Storm, G; Martin, N I; Mastrobattista, E

    2017-02-15

    Endosomolytic peptides are often coupled to drug delivery systems to enhance endosomal escape, which is crucial for the delivery of macromolecular drugs that are vulnerable to degradation in the endolysosomal pathway. Melittin is a 26 amino acid peptide derived from bee venom that has a very high membranolytic activity. However, such lytic peptides also impose a significant safety risk when applied in vivo as they often have similar activity against red blood cells and other nontarget cell membranes. Our aim is to control the membrane-disrupting capacity of these peptides in time and space by physically constraining them to a nanocarrier surface in such a way that they only become activated when delivered inside acidic endosomes. To this end, a variety of chemical approaches for the coupling of lytic peptides to liposomes via functionalized PEG-lipids were explored, including maleimide-thiol chemistry, click-chemistry, and aldehyde-hydrazide chemistry. The latter enables reversible conjugation via a hydrazone bond, allowing for release of the peptide under endosomal conditions. By carefully choosing the conjugation site and by using a pH activated analog of the melittin peptide, lytic activity toward a model membrane is completely inhibited at physiological pH. At endosomal pH the activity is restored by hydrolysis of the acid-labile hydrazone bond, releasing the peptide in its most active, free form. Furthermore, using an analogue containing a nonhydrolyzable bond as a control, it was shown that the activity observed can be completely attributed to release of the peptide, validating dynamic covalent conjugation as a suitable strategy to maintain safety during circulation.

  2. Imaging endosomes and autophagosomes in whole mammalian cells using correlative cryo-fluorescence and cryo-soft X-ray microscopy (cryo-CLXM)

    International Nuclear Information System (INIS)

    Duke, Elizabeth M.H.; Razi, Minoo; Weston, Anne; Guttmann, Peter; Werner, Stephan; Henzler, Katja; Schneider, Gerd; Tooze, Sharon A.; Collinson, Lucy M.

    2014-01-01

    Cryo-soft X-ray tomography (cryo-SXT) is a powerful imaging technique that can extract ultrastructural information from whole, unstained mammalian cells as close to the living state as possible. Subcellular organelles including the nucleus, the Golgi apparatus and mitochondria have been identified by morphology alone, due to the similarity in contrast to transmission electron micrographs. In this study, we used cryo-SXT to image endosomes and autophagosomes, organelles that are particularly susceptible to chemical fixation artefacts during sample preparation for electron microscopy. We used two approaches to identify these compartments. For early and recycling endosomes, which are accessible to externally-loaded markers, we used an anti-transferrin receptor antibody conjugated to 10 nm gold particles. For autophagosomes, which are not accessible to externally-applied markers, we developed a correlative cryo-fluorescence and cryo-SXT workflow (cryo-CLXM) to localise GFP-LC3 and RFP-Atg9. We used a stand-alone cryo-fluorescence stage in the home laboratory to localise the cloned fluorophores, followed by cryo-soft X-ray tomography at the synchrotron to analyse cellular ultrastructure. We mapped the 3D ultrastructure of the endocytic and autophagic structures, and discovered clusters of omegasomes arising from ‘hotspots’ on the ER. Thus, immunogold markers and cryo-CLXM can be used to analyse cellular processes that are inaccessible using other imaging modalities. - Highlights: • We image whole, unstained mammalian cells using cryo-soft X-ray tomography. • Endosomes are identified using a gold marker for the transferrin receptor. • A new workflow for correlative cryo-fluorescence and cryo-SXT is used to locate early autophagosomes. • Interactions between endosomes, endoplasmic reticulum and forming autophagosomes are mapped in 3D. • Multiple omegasomes are shown to form at ‘hotspots’ on the endoplasmic reticulum

  3. The small G protein Arl5 contributes to endosome-to-Golgi traffic by aiding the recruitment of the GARP complex to the Golgi

    Directory of Open Access Journals (Sweden)

    Cláudia Rosa-Ferreira

    2015-03-01

    Full Text Available The small G proteins of the Arf family play critical roles in membrane trafficking and cytoskeleton organization. However, the function of some members of the family remains poorly understood including Arl5 which is widely conserved in eukaryotes. Humans have two closely related Arl5 paralogues (Arl5a and Arl5b, and both Arl5a and Arl5b localize to the trans-Golgi with Arl5b being involved in retrograde traffic from endosomes to the Golgi apparatus. To investigate the function of Arl5, we have used Drosophila melanogaster as a model system. We find that the single Arl5 orthologue in Drosophila also localizes to the trans-Golgi, but flies lacking the Arl5 gene are viable and fertile. By using both liposome and column based affinity chromatography methods we find that Arl5 interacts with the Golgi-associated retrograde protein (GARP complex that acts in the tethering of vesicles moving from endosomes to the trans-Golgi network (TGN. In Drosophila tissues the GARP complex is partially displaced from the Golgi when Arl5 is absent, and the late endosomal compartment is enlarged. In addition, in HeLa cells GARP also becomes cytosolic upon depletion of Arl5b. These phenotypes are consistent with a role in endosome-to-Golgi traffic, but are less severe than loss of GARP itself. Thus it appears that Arl5 is one of the factors that directs the recruitment of the GARP complex to the trans-Golgi, and this function is conserved in both flies and humans.

  4. A tubular electrode for radiofrequency ablation therapy

    KAUST Repository

    Antunes, Carlos Lemos Lemos Lemos

    2012-07-06

    Purpose – Due to its good mechanical and biocompatibility characteristics, nitinol SEMS is a popular endoprothesis used for relieving stricture problems in hollow organs due to carcinomas. Besides its mechanical application, SEMS can be regarded as well as potential electrode for performing RF ablation therapy on the tumor. The purpose of this work is to perform numerical and experimental analyses in order to characterize the lesion volume induced in biological tissue using this kind of tubular electrode. Design/methodology/approach – Data concerning electrical conductivity and dimension of the damaged tissue after RF ablation procedure were obtained from ex vivo samples. Next, numerical models using 3D finite element method were obtained reassembling the conditions considered at experimentation setup and results were compared. Findings – Numerical and experimental results show that a regular volume of damaged tissue can be obtained considering this type of electrode. Also, results obtained from numerical simulation are close to those obtained by experimentation. Originality/value – SEMSs, commonly used as devices to minimize obstruction problems due to the growth of tumors, may still be considered as an active electrode for RF ablation procedures. A method considering this observation is presented in this paper. Also, numerical simulation can be regarded in this case as a tool for determining the lesion volume.

  5. Interactive Volumetry Of Liver Ablation Zones.

    Science.gov (United States)

    Egger, Jan; Busse, Harald; Brandmaier, Philipp; Seider, Daniel; Gawlitza, Matthias; Strocka, Steffen; Voglreiter, Philip; Dokter, Mark; Hofmann, Michael; Kainz, Bernhard; Hann, Alexander; Chen, Xiaojun; Alhonnoro, Tuomas; Pollari, Mika; Schmalstieg, Dieter; Moche, Michael

    2015-10-20

    Percutaneous radiofrequency ablation (RFA) is a minimally invasive technique that destroys cancer cells by heat. The heat results from focusing energy in the radiofrequency spectrum through a needle. Amongst others, this can enable the treatment of patients who are not eligible for an open surgery. However, the possibility of recurrent liver cancer due to incomplete ablation of the tumor makes post-interventional monitoring via regular follow-up scans mandatory. These scans have to be carefully inspected for any conspicuousness. Within this study, the RF ablation zones from twelve post-interventional CT acquisitions have been segmented semi-automatically to support the visual inspection. An interactive, graph-based contouring approach, which prefers spherically shaped regions, has been applied. For the quantitative and qualitative analysis of the algorithm's results, manual slice-by-slice segmentations produced by clinical experts have been used as the gold standard (which have also been compared among each other). As evaluation metric for the statistical validation, the Dice Similarity Coefficient (DSC) has been calculated. The results show that the proposed tool provides lesion segmentation with sufficient accuracy much faster than manual segmentation. The visual feedback and interactivity make the proposed tool well suitable for the clinical workflow.

  6. Interactive Volumetry Of Liver Ablation Zones

    Science.gov (United States)

    Egger, Jan; Busse, Harald; Brandmaier, Philipp; Seider, Daniel; Gawlitza, Matthias; Strocka, Steffen; Voglreiter, Philip; Dokter, Mark; Hofmann, Michael; Kainz, Bernhard; Hann, Alexander; Chen, Xiaojun; Alhonnoro, Tuomas; Pollari, Mika; Schmalstieg, Dieter; Moche, Michael

    2015-10-01

    Percutaneous radiofrequency ablation (RFA) is a minimally invasive technique that destroys cancer cells by heat. The heat results from focusing energy in the radiofrequency spectrum through a needle. Amongst others, this can enable the treatment of patients who are not eligible for an open surgery. However, the possibility of recurrent liver cancer due to incomplete ablation of the tumor makes post-interventional monitoring via regular follow-up scans mandatory. These scans have to be carefully inspected for any conspicuousness. Within this study, the RF ablation zones from twelve post-interventional CT acquisitions have been segmented semi-automatically to support the visual inspection. An interactive, graph-based contouring approach, which prefers spherically shaped regions, has been applied. For the quantitative and qualitative analysis of the algorithm’s results, manual slice-by-slice segmentations produced by clinical experts have been used as the gold standard (which have also been compared among each other). As evaluation metric for the statistical validation, the Dice Similarity Coefficient (DSC) has been calculated. The results show that the proposed tool provides lesion segmentation with sufficient accuracy much faster than manual segmentation. The visual feedback and interactivity make the proposed tool well suitable for the clinical workflow.

  7. L-Type Voltage-Gated Ca2+ Channels Regulate Synaptic-Activity-Triggered Recycling Endosome Fusion in Neuronal Dendrites

    Directory of Open Access Journals (Sweden)

    Brian G. Hiester

    2017-11-01

    Full Text Available The repertoire and abundance of proteins displayed on the surface of neuronal dendrites are tuned by regulated fusion of recycling endosomes (REs with the dendritic plasma membrane. While this process is critical for neuronal function and plasticity, how synaptic activity drives RE fusion remains unexplored. We demonstrate a multistep fusion mechanism that requires Ca2+ from distinct sources. NMDA receptor Ca2+ initiates RE fusion with the plasma membrane, while L-type voltage-gated Ca2+ channels (L-VGCCs regulate whether fused REs collapse into the membrane or reform without transferring their cargo to the cell surface. Accordingly, NMDA receptor activation triggered AMPA-type glutamate receptor trafficking to the dendritic surface in an L-VGCC-dependent manner. Conversely, potentiating L-VGCCs enhanced AMPA receptor surface expression only when NMDA receptors were also active. Thus L-VGCCs play a role in tuning activity-triggered surface expression of key synaptic proteins by gating the mode of RE fusion.

  8. The Human ABCG1 Transporter Mobilizes Plasma Membrane and Late Endosomal Non-Sphingomyelin-Associated-Cholesterol for Efflux and Esterification

    Directory of Open Access Journals (Sweden)

    Edward B. Neufeld

    2014-12-01

    Full Text Available We have previously shown that GFP-tagged human ABCG1 on the plasma membrane (PM and in late endosomes (LE mobilizes sterol on both sides of the membrane lipid bilayer, thereby increasing cellular cholesterol efflux to lipid surfaces. In the present study, we examined ABCG1-induced changes in membrane cholesterol distribution, organization, and mobility. ABCG1-GFP expression increased the amount of mobile, non-sphingomyelin(SM-associated cholesterol at the PM and LE, but not the amount of SM-associated-cholesterol or SM. ABCG1-mobilized non-SM-associated-cholesterol rapidly cycled between the PM and LE and effluxed from the PM to extracellular acceptors, or, relocated to intracellular sites of esterification. ABCG1 increased detergent-soluble pools of PM and LE cholesterol, generated detergent-resistant, non-SM-associated PM cholesterol, and increased resistance to both amphotericin B-induced (cholesterol-mediated and lysenin-induced (SM-mediated cytolysis, consistent with altered organization of both PM cholesterol and SM. ABCG1 itself resided in detergent-soluble membrane domains. We propose that PM and LE ABCG1 residing at the phase boundary between ordered (Lo and disordered (Ld membrane lipid domains alters SM and cholesterol organization thereby increasing cholesterol flux between Lo and Ld, and hence, the amount of cholesterol available for removal by acceptors on either side of the membrane bilayer for either efflux or esterification.

  9. The Cdc42 guanine nucleotide exchange factor FGD6 coordinates cell polarity and endosomal membrane recycling in osteoclasts.

    Science.gov (United States)

    Steenblock, Charlotte; Heckel, Tobias; Czupalla, Cornelia; Espírito Santo, Ana Isabel; Niehage, Christian; Sztacho, Martin; Hoflack, Bernard

    2014-06-27

    The initial step of bone digestion is the adhesion of osteoclasts onto bone surfaces and the assembly of podosomal belts that segregate the bone-facing ruffled membrane from other membrane domains. During bone digestion, membrane components of the ruffled border also need to be recycled after macropinocytosis of digested bone materials. How osteoclast polarity and membrane recycling are coordinated remains unknown. Here, we show that the Cdc42-guanine nucleotide exchange factor FGD6 coordinates these events through its Src-dependent interaction with different actin-based protein networks. At the plasma membrane, FGD6 couples cell adhesion and actin dynamics by regulating podosome formation through the assembly of complexes comprising the Cdc42-interactor IQGAP1, the Rho GTPase-activating protein ARHGAP10, and the integrin interactors Talin-1/2 or Filamin A. On endosomes and transcytotic vesicles, FGD6 regulates retromer-dependent membrane recycling through its interaction with the actin nucleation-promoting factor WASH. These results provide a mechanism by which a single Cdc42-exchange factor controlling different actin-based processes coordinates cell adhesion, cell polarity, and membrane recycling during bone degradation. © 2014 by The American Society for Biochemistry and Molecular Biology, Inc.

  10. Enrichment of Phosphatidylethanolamine in Viral Replication Compartments via Co-opting the Endosomal Rab5 Small GTPase by a Positive-Strand RNA Virus.

    Directory of Open Access Journals (Sweden)

    Kai Xu

    2016-10-01

    Full Text Available Positive-strand RNA viruses build extensive membranous replication compartments to support replication and protect the virus from antiviral responses by the host. These viruses require host factors and various lipids to form viral replication complexes (VRCs. The VRCs built by Tomato bushy stunt virus (TBSV are enriched with phosphatidylethanolamine (PE through a previously unknown pathway. To unravel the mechanism of PE enrichment within the TBSV replication compartment, in this paper, the authors demonstrate that TBSV co-opts the guanosine triphosphate (GTP-bound active form of the endosomal Rab5 small GTPase via direct interaction with the viral replication protein. Deletion of Rab5 orthologs in a yeast model host or expression of dominant negative mutants of plant Rab5 greatly decreases TBSV replication and prevents the redistribution of PE to the sites of viral replication. We also show that enrichment of PE in the viral replication compartment is assisted by actin filaments. Interestingly, the closely related Carnation Italian ringspot virus, which replicates on the boundary membrane of mitochondria, uses a similar strategy to the peroxisomal TBSV to hijack the Rab5-positive endosomes into the viral replication compartments. Altogether, usurping the GTP-Rab5-positive endosomes allows TBSV to build a PE-enriched viral replication compartment, which is needed to support peak-level replication. Thus, the Rab family of small GTPases includes critical host factors assisting VRC assembly and genesis of the viral replication compartment.

  11. The functional interplay of Rab11, FIP3 and Rho proteins on the endosomal recycling pathway controls cell shape and symmetry.

    Science.gov (United States)

    Bouchet, Jérôme; McCaffrey, Mary W; Graziani, Andrea; Alcover, Andrés

    2018-07-04

    Several families of small GTPases regulate a variety of fundamental cellular processes, encompassing growth factor signal transduction, vesicular trafficking and control of the cytoskeleton. Frequently, their action is hierarchical and complementary, but much of the detail of their functional interactions remains to be clarified. It is well established that Rab family members regulate a variety of intracellular vesicle trafficking pathways. Moreover, Rho family GTPases are pivotal for the control of the actin and microtubule cytoskeleton. However, the interplay between these 2 types of GTPases has been rarely reported. We discuss here our recent findings showing that Rab11, a key regulator of endosomal recycling, and Rac1, a central actin cytoskeleton regulator involved in lamellipodium formation and cell migration, interplay on endosomes through the Rab11 effector FIP3. In the context of the rapidly reactive T lymphocytes, Rab11-Rac1 endosomal functional interplay is important to control cell shape changes and cell symmetry during lymphocyte spreading and immunological synapse formation and ultimately modulate T cell activation.

  12. The Vici Syndrome Protein EPG5 Is a Rab7 Effector that Determines the Fusion Specificity of Autophagosomes with Late Endosomes/Lysosomes.

    Science.gov (United States)

    Wang, Zheng; Miao, Guangyan; Xue, Xue; Guo, Xiangyang; Yuan, Chongzhen; Wang, Zhaoyu; Zhang, Gangming; Chen, Yingyu; Feng, Du; Hu, Junjie; Zhang, Hong

    2016-09-01

    Mutations in the human autophagy gene EPG5 cause the multisystem disorder Vici syndrome. Here we demonstrated that EPG5 is a Rab7 effector that determines the fusion specificity of autophagosomes with late endosomes/lysosomes. EPG5 is recruited to late endosomes/lysosomes by direct interaction with Rab7 and the late endosomal/lysosomal R-SNARE VAMP7/8. EPG5 also binds to LC3/LGG-1 (mammalian and C. elegans Atg8 homolog, respectively) and to assembled STX17-SNAP29 Qabc SNARE complexes on autophagosomes. EPG5 stabilizes and facilitates the assembly of STX17-SNAP29-VAMP7/8 trans-SNARE complexes, and promotes STX17-SNAP29-VAMP7-mediated fusion of reconstituted proteoliposomes. Loss of EPG5 activity causes abnormal fusion of autophagosomes with various endocytic vesicles, in part due to elevated assembly of STX17-SNAP25-VAMP8 complexes. SNAP25 knockdown partially suppresses the autophagy defect caused by EPG5 depletion. Our study reveals that EPG5 is a Rab7 effector involved in autophagosome maturation, providing insight into the molecular mechanism underlying Vici syndrome. Copyright © 2016 Elsevier Inc. All rights reserved.

  13. Modeling CO2 Laser Ablative Impulse with Polymers

    International Nuclear Information System (INIS)

    Sinko, John E.; Phipps, Claude R.; Sasoh, Akihiro

    2010-01-01

    Laser ablation vaporization models have usually ignored the spatial dependence of the laser beam. Here, we consider effects from modeling using a Gaussian beam for both photochemical and photothermal conditions. The modeling results are compared to experimental and literature data for CO 2 laser ablation of the polymer polyoxymethylene under vacuum, and discussed in terms of the ablated mass areal density and momentum coupling coefficient. Extending the scope of discussion, laser ablative impulse generation research has lacked a cohesive strategy for linking the vaporization and plasma regimes. Existing models, mostly formulated for ultraviolet laser systems or metal targets, appear to be inappropriate or impractical for applications requiring CO 2 laser ablation of polymers. A recently proposed method for linking the vaporization and plasma regimes for analytical modeling is addressed here along with the implications of its use. Key control parameters are considered, along with the major propulsion parameters needed for laser ablation propulsion modeling.

  14. Transarterial ethanol ablation for sporadic and non-hemorrhaging angiomyolipoma in the kidney

    International Nuclear Information System (INIS)

    Takebayashi, Shigeo; Horikawa, Ayumi; Arai, Mito; Iso, Shinichiroh; Noguchi, Kazumi

    2009-01-01

    Purpose: We evaluated the efficacy and side effects of transarterial ethanol ablation in sporadic and non-hemorrhaging angiomyolipomas (AMLs) in the kidney. Material and Methods: A total of 10 patients with solitary and sporadic AMLs underwent selective transarterial absolute ethanol ablation for prophylaxis against hemorrhage. We confirmed the ratio areas of tumor vessel on angiogram, those of infraction on post-ablation computed tomography (CT) and those of tumor reduction in a 3-, 6- and 12-month follow-up CT. Results: Once or twice a single infusion of 1 or 2 ml absolute ethanol achieved in a total occlusion of 22 feeding arteries which consisted of 7 proximal interlobar arteries, 12 distal interlobar arteries and 3 renal capsular arteries. Nontarget occlusion did not occur by ethanol reflux in any cases but occurred causing spasms provoked by repeated inflation and deflation of the balloon in one case. Total occlusion of tumor vessels was observed in 7 patients and 92-95% occlusion in 3. Ethanol ablation produced 1.8-22.5% (mean 8.4 ± 6.8%) areas of infarctions but the outcome was not serious in all cases. Mean percentage areas of tumor reduction were 29.4 ± 10.6% in a 3-month follow-up, 45.7 ± 11.9% in a 6-month and 59.3 ± 11.5% in a 12-month follow-up. Conclusions: Absolute ethanol ablation for sporadic and non-hemorrhaging AML is safe and effective in reducing majority of tumor area in a 1-year follow-up.

  15. Innovative techniques for image-guided ablation of benign thyroid nodules: Combined ethanol and radiofrequency ablation

    Energy Technology Data Exchange (ETDEWEB)

    Park, Hye Sun; Baek, Jung Hwan; Choi, Young Jun; Lee, Jeong Hyun [Dept. of Radiology and Research Institute of Radiology, University of Ulsan College of Medicine, Asan Medical Center, Seoul (Korea, Republic of)

    2017-06-15

    In the treatment of benign thyroid nodules, ethanol ablation (EA), and radiofrequency ablation (RFA) have been suggested for cystic and solid thyroid nodules, respectively. Although combining these ablation techniques may be effective, no guidelines for or reviews of the combination have been published. Currently, there are three ways of combining EA and RFA: additional RFA is effective for treatment of incompletely resolved symptoms and solid residual portions of a thyroid nodule after EA. Additional EA can be performed for the residual unablated solid portion of a nodule after RFA if it is adjacent to critical structures (e.g., trachea, esophagus, and recurrent laryngeal nerve). In the concomitant procedure, ethanol is injected to control venous oozing after aspiration of cystic fluid prior to RFA of the remaining solid nodule.

  16. Convergent ablation measurements of plastic ablators in gas-filled rugby hohlraums on OMEGA

    Science.gov (United States)

    Casner, A.; Jalinaud, T.; Masse, L.; Galmiche, D.

    2015-10-01

    Indirect-drive implosions experiments were conducted on the Omega Laser Facility to test the performance of uniformly doped plastic ablators for Inertial Confinement Fusion. The first convergent ablation measurements in gas-filled rugby hohlraums are reported. Ignition relevant limb velocities in the range from 150 to 300 μm .n s-1 have been reached by varying the laser drive energy and the initial capsule aspect ratio. The measured capsule trajectory and implosion velocity are in good agreement with 2D integrated simulations and a zero-dimensional modeling of the implosions. We demonstrate experimentally the scaling law for the maximum implosion velocity predicted by the improved rocket model [Y. Saillard, Nucl. Fusion 46, 1017 (2006)] in the high-ablation regime case.

  17. Ablation by ultrashort laser pulses: Atomistic and thermodynamic analysis of the processes at the ablation threshold

    International Nuclear Information System (INIS)

    Upadhyay, Arun K.; Inogamov, Nail A.; Rethfeld, Baerbel; Urbassek, Herbert M.

    2008-01-01

    Ultrafast laser irradiation of solids may ablate material off the surface. We study this process for thin films using molecular-dynamics simulation and thermodynamic analysis. Both metals and Lennard-Jones (LJ) materials are studied. We find that despite the large difference in thermodynamical properties between these two classes of materials--e.g., for aluminum versus LJ the ratio T c /T tr of critical to triple-point temperature differs by more than a factor of 4--the values of the ablation threshold energy E abl normalized to the cohesion energy, ε abl =E abl /E coh , are surprisingly universal: all are near 0.3 with ±30% scattering. The difference in the ratio T c /T tr means that for metals the melting threshold ε m is low, ε m abl , while for LJ it is high, ε m >ε abl . This thermodynamical consideration gives a simple explanation for the difference between metals and LJ. It explains why despite the universality in ε abl , metals thermomechanically ablate always from the liquid state. This is opposite to LJ materials, which (near threshold) ablate from the solid state. Furthermore, we find that immediately below the ablation threshold, the formation of large voids (cavitation) in the irradiated material leads to a strong temporary expansion on a very slow time scale. This feature is easily distinguished from the acoustic oscillations governing the material response at smaller intensities, on the one hand, and the ablation occurring at larger intensities, on the other hand. This finding allows us to explain the puzzle of huge surface excursions found in experiments at near-threshold laser irradiation

  18. Ablation and optical third-order nonlinearities in Ag nanoparticles

    Directory of Open Access Journals (Sweden)

    Carlos Torres-Torres

    2010-11-01

    Full Text Available Carlos Torres-Torres1, Néstor Peréa-López2, Jorge Alejandro Reyes-Esqueda3, Luis Rodríguez-Fernández3, Alejandro Crespo-Sosa3, Juan Carlos Cheang-Wong3, Alicia Oliver31Section of Graduate Studies and Research, School of Mechanical and Electrical Engineering, National Polytechnic Institute, Zacatenco, Distrito Federal, Mexico; 2Laboratory for Nanoscience and Nanotechnology Research and Advanced Materials Department, IPICYT, Camino a la Presa San Jose, San Luis Potosi, Mexico; 3Instituto de Física, Universidad Nacional Autónoma de México, A.P. 20-364, México, D.F. 01000, MéxicoAbstract: The optical damage associated with high intensity laser excitation of silver nanoparticles (NPs was studied. In order to investigate the mechanisms of optical nonlinearity of a nanocomposite and their relation with its ablation threshold, a high-purity silica sample implanted with Ag ions was exposed to different nanosecond and picosecond laser irradiations. The magnitude and sign of picosecond refractive and absorptive nonlinearities were measured near and far from the surface plasmon resonance (SPR of the Ag NPs with a self-diffraction technique. Saturable optical absorption and electronic polarization related to self-focusing were identified. Linear absorption is the main process involved in nanosecond laser ablation, but nonlinearities are important for ultrashort picosecond pulses when the absorptive process become significantly dependent on the irradiance. We estimated that near the resonance, picosecond intraband transitions allow an expanded distribution of energy among the NPs, in comparison to the energy distribution resulting in a case of far from resonance, when the most important absorption takes place in silica. We measured important differences in the ablation threshold and we estimated that the high selectiveness of the SPR of Ag NPs as well as their corresponding optical nonlinearities can be strongly significant for laser

  19. Enhanced Radiofrequency Ablation With Magnetically Directed Metallic Nanoparticles.

    Science.gov (United States)

    Nguyen, Duy T; Tzou, Wendy S; Zheng, Lijun; Barham, Waseem; Schuller, Joseph L; Shillinglaw, Benjamin; Quaife, Robert A; Sauer, William H

    2016-05-01

    Remote heating of metal located near a radiofrequency ablation source has been previously demonstrated. Therefore, ablation of cardiac tissue treated with metallic nanoparticles may improve local radiofrequency heating and lead to larger ablation lesions. We sought to evaluate the effect of magnetic nanoparticles on tissue sensitivity to radiofrequency energy. Ablation was performed using an ablation catheter positioned with 10 g of force over prepared ex vivo specimens. Tissue temperatures were measured and lesion volumes were acquired. An in vivo porcine thigh model was used to study systemically delivered magnetically guided iron oxide (FeO) nanoparticles during radiofrequency application. Magnetic resonance imaging and histological staining of ablated tissue were subsequently performed as a part of ablation lesion analysis. Ablation of ex vivo myocardial tissue treated with metallic nanoparticles resulted in significantly larger lesions with greater impedance changes and evidence of increased thermal conductivity within the tissue. Magnet-guided localization of FeO nanoparticles within porcine thigh preps was demonstrated by magnetic resonance imaging and iron staining. Irrigated ablation in the regions with greater FeO, after FeO infusion and magnetic guidance, created larger lesions without a greater incidence of steam pops. Metal nanoparticle infiltration resulted in significantly larger ablation lesions with altered electric and thermal conductivity. In vivo magnetic guidance of FeO nanoparticles allowed for facilitated radiofrequency ablation without direct infiltration into the targeted tissue. Further research is needed to assess the clinical applicability of this ablation strategy using metallic nanoparticles for the treatment of cardiac arrhythmias. © 2016 American Heart Association, Inc.

  20. Design of Ablation Test Device for Brick Coating of Gun

    Science.gov (United States)

    shirui, YAO; yongcai, CHEN; fei, WANG; jianxin, ZHAO

    2018-03-01

    As a result of the live ammunition test conditions, the barrel resistance of the barrel coating has high cost, time consuming, low efficiency and high test site requirements. This article designed a simple, convenient and efficient test device. Through the internal trajectory calculation by Matlab, the ablation environment produced by the ablation test device has achieved the expected effect, which is consistent with the working condition of the tube in the launching state, which can better reflect the ablation of the coating.

  1. A 6-year review of the outcome of endometrial ablation.

    Science.gov (United States)

    Tsaltas, J; Taylor, N; Healey, M

    1998-02-01

    In June, 1995 a postal questionnaire was distributed to all 232 women who had an endometrial ablation at Monash Medical Centre between July, 1989 and December, 1994. Data was analyzed from the 149 who responded. Length of follow-up ranged from 6 months to 6 years 6 months. Of these 78% were satisfied with their ablation and 84% found their menses to be lighter or to have stopped. The repeat ablation rate was 13% and the hysterectomy rate was 17%.

  2. Measurement of intrahepatic pressure during radiofrequency ablation in porcine liver.

    Science.gov (United States)

    Kawamoto, Chiaki; Yamauchi, Atsushi; Baba, Yoko; Kaneko, Keiko; Yakabi, Koji

    2010-04-01

    To identify the most effective procedures to avoid increased intrahepatic pressure during radiofrequency ablation, we evaluated different ablation methods. Laparotomy was performed in 19 pigs. Intrahepatic pressure was monitored using an invasive blood pressure monitor. Radiofrequency ablation was performed as follows: single-step standard ablation; single-step at 30 W; single-step at 70 W; 4-step at 30 W; 8-step at 30 W; 8-step at 70 W; and cooled-tip. The array was fully deployed in single-step methods. In the multi-step methods, the array was gradually deployed in four or eight steps. With the cooled-tip, ablation was performed by increasing output by 10 W/min, starting at 40 W. Intrahepatic pressure was as follows: single-step standard ablation, 154.5 +/- 30.9 mmHg; single-step at 30 W, 34.2 +/- 20.0 mmHg; single-step at 70 W, 46.7 +/- 24.3 mmHg; 4-step at 30 W, 42.3 +/- 17.9 mmHg; 8-step at 30 W, 24.1 +/- 18.2 mmHg; 8-step at 70 W, 47.5 +/- 31.5 mmHg; and cooled-tip, 114.5 +/- 16.6 mmHg. The radiofrequency ablation-induced area was spherical with single-step standard ablation, 4-step at 30 W, and 8-step at 30 W. Conversely, the ablated area was irregular with single-step at 30 W, single-step at 70 W, and 8-step at 70 W. The ablation time was significantly shorter for the multi-step method than for the single-step method. Increased intrahepatic pressure could be controlled using multi-step methods. From the shapes of the ablation area, 30-W 8-step expansions appear to be most suitable for radiofrequency ablation.

  3. Comparative evaluation of diode laser ablation and surgical stripping technique for gingival depigmentation: A clinical and immunohistochemical study.

    Science.gov (United States)

    Bakutra, Gaurav; Shankarapillai, Rajesh; Mathur, Lalit; Manohar, Balaji

    2017-01-01

    There are various treatment modalities to remove the black patches of melanin pigmentation. The aim of the study is to clinically compare the diode laser ablation and surgical stripping technique for gingival depigmentation and to evaluate their effect on the histological changes in melanocyte activity. A total of 40 sites of 20 patients with bilateral melanin hyperpigmentation were treated with the surgical stripping and diode laser ablation technique. Change in Hedin index score, change in area of pigmentation using image analyzing software, pain perception, patient preference of treatment were recorded. All 40 sites were selected for immunohistochemical analysis using HMB-45 immunohistochemical marker. At 12 months post-operative visit, in all sites, repigmentation was observed with different grades of Hedin index. Paired t -test, analysis of variance, and Chi-square tests were used for statistical analysis. Repigmentation in surgical stripping is significantly lesser compared to laser ablation. Lesser numbers of melanocytes were found on immunohistological examination at 12 months postoperatively. Comparison for patient preference and pain indices give statistically significant values for diode laser techniques. Gingival hyperpigmentation is effectively managed by diode laser ablation technique and surgical stripping method. In this study, surgical stripping technique found to be better compared to diode laser ablation.

  4. Fiber-Optic Temperature and Pressure Sensors Applied to Radiofrequency Thermal Ablation in Liver Phantom: Methodology and Experimental Measurements

    Directory of Open Access Journals (Sweden)

    Daniele Tosi

    2015-01-01

    Full Text Available Radiofrequency thermal ablation (RFA is a procedure aimed at interventional cancer care and is applied to the treatment of small- and midsize tumors in lung, kidney, liver, and other tissues. RFA generates a selective high-temperature field in the tissue; temperature values and their persistency are directly related to the mortality rate of tumor cells. Temperature measurement in up to 3–5 points, using electrical thermocouples, belongs to the present clinical practice of RFA and is the foundation of a physical model of the ablation process. Fiber-optic sensors allow extending the detection of biophysical parameters to a vast plurality of sensing points, using miniature and noninvasive technologies that do not alter the RFA pattern. This work addresses the methodology for optical measurement of temperature distribution and pressure using four different fiber-optic technologies: fiber Bragg gratings (FBGs, linearly chirped FBGs (LCFBGs, Rayleigh scattering-based distributed temperature system (DTS, and extrinsic Fabry-Perot interferometry (EFPI. For each instrument, methodology for ex vivo sensing, as well as experimental results, is reported, leading to the application of fiber-optic technologies in vivo. The possibility of using a fiber-optic sensor network, in conjunction with a suitable ablation device, can enable smart ablation procedure whereas ablation parameters are dynamically changed.

  5. Efficiency of ablative plasma energy transfer into a massive aluminum target using different atomic number ablators

    Czech Academy of Sciences Publication Activity Database

    Kasperczuk, A.; Pisarczyk, T.; Chodukowski, T.; Kalinowska, Z.; Stepniewski, W.; Jach, K.; Swierczynski, R.; Renner, Oldřich; Šmíd, Michal; Ullschmied, Jiří; Cikhart, J.; Klír, D.; Kubeš, P.; Řezáč, K.; Krouský, Eduard; Pfeifer, Miroslav; Skála, Jiří

    2015-01-01

    Roč. 33, č. 3 (2015), s. 379-386 ISSN 0263-0346 R&D Projects: GA MŠk ED1.1.00/02.0061; GA MŠk(CZ) LD14089 EU Projects: European Commission(XE) 284464 - LASERLAB-EUROPE Grant - others:ELI Beamlines(XE) CZ.1.05/1.1.00/02.0061; AVČR(CZ) M100101208 Institutional support: RVO:68378271 ; RVO:61389021 Keywords : ablator atomic number * crater volume * laser energy transfer * plasma ablative pressure Subject RIV: BL - Plasma and Gas Discharge Physics; BH - Optics, Masers, Lasers (UFP-V) Impact factor: 1.649, year: 2015

  6. Efficacy and safety of ultrasound-guided radiofrequency ablation of hyperplastic parathyroid gland for secondary hyperparathyroidism associated with chronic kidney disease.

    Science.gov (United States)

    Peng, Chengzhong; Zhang, Zhengxian; Liu, Jibin; Chen, Hongyu; Tu, Xiao; Hu, Rihong; Ni, Jun; Weng, Ning; Pang, Haisu; Xue, Zhengmei

    2017-03-01

    The purpose of this study was to determine if ultrasound-guided radiofrequency ablation (RFA) of hyperplastic parathyroid glands could be used to treat secondary hyperparathyroidism (HPT) in patients with chronic kidney disease. RFA of the hyperplastic parathyroid glands was performed in 34 patients with secondary HPT. Intact parathyroid hormone (iPTH), calcium, and phosphorus were measured. The outcome was based on the ablation extent (ie, 4, 3, and 1-2 glands). The iPTH, calcium, and phosphorus levels decreased in all groups after RFA. One year after ablation, these parameters remained significantly lower in the 4-gland ablation group compared with the 3-gland and 1 to 2-gland groups. The same tendency was observed for the symptom score. The iPTH levels of secondary HPT is feasible in selected patients. © 2016 Wiley Periodicals, Inc. Head Neck 39: 564-571, 2017. © 2016 Wiley Periodicals, Inc.

  7. CT-guided radiofrequency tumor ablation in children

    International Nuclear Information System (INIS)

    Botsa, Evanthia; Poulou, Loukia S.; Koundouraki, Antonia; Thanos, Loukas; Koutsogiannis, Ioannis; Ziakas, Panayiotis D.; Alexopoulou, Efthimia

    2014-01-01

    Image-guided radiofrequency ablation is a well-accepted technique of interventional oncology in adults. To evaluate the efficacy and safety of CT-guided radiofrequency ablation as a minimally invasive treatment for metastatic neoplasms in children. A total of 15 radiofrequency ablation sessions were performed in 12 children and young adults (median age 9.5; range 5-18 years) with metastatic malignancies. Seven children and young adults had secondary hepatic lesions, three had pulmonary and two had bone lesions. Radiofrequency ablation was performed under conscious sedation. The median lesion size was 1.7 cm (range 1.3-2.8 cm). The median time for ablation was 8 min (range 7-10 min). Radiofrequency procedures were technically successful in all tumors. Postablation imaging immediately after, and 1 month and 3 months after radiofrequency ablation showed total necrosis in all patients. At 6-month follow-up, three patients (all with lesion size >2 cm) had local recurrence and underwent a second radiofrequency ablation session. At 2-year follow-up no patient had recurrence of the treated tumor. Post-ablation syndrome occurred in four children. No major complication occurred. CT-guided radiofrequency tumor ablation was safe and efficient for palliative treatment in our cohort of patients. (orig.)

  8. Image-guided radiofrequency ablation of renal cell carcinoma

    International Nuclear Information System (INIS)

    Boss, Andreas; Clasen, Stephan; Pereira, Philippe L.; Kuczyk, Markus; Schick, Fritz

    2007-01-01

    The incidence of renal cell carcinoma is rising with the increased number of incidental detection of small tumours. During the past few years, percutaneous imaging-guided radiofrequency ablation has evolved as a minimally invasive treatment of small unresectable renal tumours offering reduced patient morbidity and overall health care costs. In radiofrequency ablation, thermal energy is deposited into a targeted tumour by means of a radiofrequency applicator. In recent studies, radiofrequency ablation was shown to be an effective and safe modality for local destruction of renal cell carcinoma. Radiofrequency applicator navigation can be performed via ultrasound, computed tomography or magnetic resonance guidance; however, ultrasound seems less favourable because of the absence of monitoring capabilities during ablation. On-line monitoring of treatment outcome can only be performed with magnetic resonance imaging giving the possibility of eventual applicator repositioning to ablate visible residual tumour tissue. Long-term follow-up is crucial to assess completeness of tumour ablation. New developments in ablation technology and radiological equipment will further increase the indication field for radiofrequency ablation of renal cell carcinoma. Altogether, radiofrequency ablation seems to be a promising new modality for the minimally invasive treatment of renal cell carcinoma, which was demonstrated to exhibit high short-term effectiveness. (orig.)

  9. Osteoid Osteoma: Experience with Laser- and Radiofrequency-Induced Ablation

    International Nuclear Information System (INIS)

    Gebauer, Bernhard; Tunn, Per-Ulf; Gaffke, Gunnar; Melcher, Ingo; Felix, Roland; Stroszczynski, Christian

    2006-01-01

    The purpose of this study was to analyze the clinical outcome of osteoid osteoma treated by thermal ablation after drill opening. A total of 17 patients and 20 procedures were included. All patients had typical clinical features (age, pain) and a typical radiograph showing a nidus. In 5 cases, additional histological specimens were acquired. After drill opening of the osteoid osteoma nidus, 12 thermal ablations were induced by laser interstitial thermal therapy (LITT) (9F Power-Laser-Set; Somatex, Germany) and 8 ablations by radiofrequency ablation (RFA) (RITA; StarBurst, USA). Initial clinical success with pain relief has been achieved in all patients after the first ablation. Three patients had an osteoid osteoma recurrence after 3, 9, and 10 months and were successfully re-treated by thermal ablation. No major complication and one minor complication (sensible defect) were recorded. Thermal ablation is a safe and minimally invasive therapy option for osteoid osteoma. Although the groups are too small for a comparative analysis, we determined no difference between laser- and radiofrequency-induced ablation in clinical outcome after ablation

  10. A thermal model for nanosecond pulsed laser ablation of aluminum

    Directory of Open Access Journals (Sweden)

    Yu Zhang

    2017-07-01

    Full Text Available In order to simulate the nanosecond pulsed laser ablation of aluminum, a novel model was presented for the target ablation and plume expansion. The simulation of the target ablation was based on one-dimensional heat conduction, taking into account temperature dependent material properties, phase transition, dielectric transition and phase explosion. While the simulation of the plume expansion was based on one-dimensional gas-dynamical equation, taking into account ionization, plume absorption and shielding. By coupling the calculations of the target ablation and plume expansion, the characteristics of the target and plume were obtained. And the calculated results were in good agreement with the experimental data, in terms of ablation threshold and depth within the fluence range of the tested laser. Subsequently, investigations were carried out to analyze the mechanisms of nanosecond pulsed laser ablation. The calculated results showed that the maximum surface temperature remained at about 90% of the critical temperature (0.9Tc due to phase explosion. Moreover, the plume shielding has significant effects on the laser ablation, and the plume shielding proportion increase as the laser fluence increasing. The ambient pressure belows 100 Pa is more suitable for laser ablation, which can obtained larger ablation depth.

  11. Using Target Ablation for Ion Beam Quality Improvement

    International Nuclear Information System (INIS)

    Zhao Shuan; Chen Jia-Er; Lin Chen; Ma Wen-Jun; Yan Xue-Qing; Wang Jun-Jie

    2016-01-01

    During the laser foil interaction, the output ion beam quality including the energy spread and beam divergence can be improved by the target ablation, due to the direct laser acceleration (DLA) electrons generated in the ablation plasma. The acceleration field established at the target rear by these electrons, which is highly directional and triangle-envelope, is helpful for the beam quality. With the help of the target ablation, both the beam divergence and energy spread will be reduced. If the ablation is more sufficient, the impact of DLA-electron-caused field will be strengthened, and the beam quality will be better, confirmed by the particle-in-cell simulation. (paper)

  12. Development of laser ablation plasma by anisotropic self-radiation

    Directory of Open Access Journals (Sweden)

    Ohnishi Naofumi

    2013-11-01

    Full Text Available We have proposed a method for reproducing an accurate solution of low-density ablation plasma by properly treating anisotropic radiation. Monte-Carlo method is employed for estimating Eddington tensor with limited number of photon samples in each fluid time step. Radiation field from ablation plasma is significantly affected by the anisotropic Eddington tensor. Electron temperature around the ablation surface changes with the radiation field and is responsible for the observed emission. An accurate prediction of the light emission from the laser ablation plasma requires a careful estimation of the anisotropic radiation field.

  13. CT-guided radiofrequency tumor ablation in children

    Energy Technology Data Exchange (ETDEWEB)

    Botsa, Evanthia [National and Kapodistrian University of Athens, First Pediatric Clinic, Agia Sofia Children' s Hospital, Athens (Greece); Poulou, Loukia S.; Koundouraki, Antonia; Thanos, Loukas [Sotiria General Hospital for Chest Diseases, Department of Medical Imaging and Interventional Radiology, Athens (Greece); Koutsogiannis, Ioannis [General Military Hospital NIMTS, Department of Medical Imaging, Athens (Greece); Ziakas, Panayiotis D. [Warren Alpert Medical School of Brown University Rhode Island Hospital, Division of Infectious Diseases, Providence, RI (United States); Alexopoulou, Efthimia [Attikon University Hospital, Second Department of Radiology, Athens University School of Medicine, Athens (Greece)

    2014-11-15

    Image-guided radiofrequency ablation is a well-accepted technique of interventional oncology in adults. To evaluate the efficacy and safety of CT-guided radiofrequency ablation as a minimally invasive treatment for metastatic neoplasms in children. A total of 15 radiofrequency ablation sessions were performed in 12 children and young adults (median age 9.5; range 5-18 years) with metastatic malignancies. Seven children and young adults had secondary hepatic lesions, three had pulmonary and two had bone lesions. Radiofrequency ablation was performed under conscious sedation. The median lesion size was 1.7 cm (range 1.3-2.8 cm). The median time for ablation was 8 min (range 7-10 min). Radiofrequency procedures were technically successful in all tumors. Postablation imaging immediately after, and 1 month and 3 months after radiofrequency ablation showed total necrosis in all patients. At 6-month follow-up, three patients (all with lesion size >2 cm) had local recurrence and underwent a second radiofrequency ablation session. At 2-year follow-up no patient had recurrence of the treated tumor. Post-ablation syndrome occurred in four children. No major complication occurred. CT-guided radiofrequency tumor ablation was safe and efficient for palliative treatment in our cohort of patients. (orig.)

  14. Second-generation endometrial ablation technologies: the hot liquid balloons.

    Science.gov (United States)

    Vilos, George A; Edris, Fawaz

    2007-12-01

    Hysteroscopic endometrial ablation (HEA) was introduced in the 1980s to treat menorrhagia. Its use required additional training, surgical expertise and specialized equipment to minimize emergent complications such as uterine perforations, thermal injuries and excessive fluid absorption. To overcome these difficulties and concerns, thermal balloon endometrial ablation (TBEA) was introduced in the 1990s. Four hot liquid balloons have been introduced into clinical practice. All systems consist of a catheter (4-10mm diameter), a silicone balloon and a control unit. Liquids used to inflate the balloons include internally heated dextrose in water (ThermaChoice, 87 degrees C), and externally heated glycine (Cavaterm, 78 degrees C), saline (Menotreat, 85 degrees ) and glycerine (Thermablate, 173 degrees C). All balloons require pressurization from 160 to 240 mmHg for treatment cycles of 2 to 10 minutes. Prior to TBEA, preoperative endometrial thinning, including suction curettage, is optional. Several RCTs and cohort studies indicate that the advantages of TBEA include portability, ease of use and short learning curve. In addition, small diameter catheters requiring minimal cervical dilatation (5-7 mm) and short duration of treatment cycles (2-8 min) allow treatment under minimal analgesia/anesthesia requirements in a clinic setting. Following TBEA serious adverse events, including thermal injuries to viscera have been experienced. To minimize such injuries some surgeons advocate the use of routine post-dilatation hysteroscopy and/or ultrasonography to confirm correct intrauterine placement of the balloon prior to initiating the treatment cycle. After 10 years of clinical practice, TBEA is thought to be the preferred first-line surgical treatment of menorrhagia in appropriately selected candidates. Economic modeling also suggested that TBEA may be more cost-effective than HEA.

  15. Junctional rhythm occurring during AV nodal reentrant tachycardia ablation, is it different among Egyptians?

    Directory of Open Access Journals (Sweden)

    Ayman M. Abdel Moteleb

    2013-12-01

    Conclusion: Junctional rhythm is a sensitive predictor of successful ablation. The pattern of JR is a useful predictor of successful ablation. Egyptian population has distinctive patterns of JR during AVNRT ablation.

  16. CT-guided Bipolar and Multipolar Radiofrequency Ablation (RF Ablation) of Renal Cell Carcinoma: Specific Technical Aspects and Clinical Results

    International Nuclear Information System (INIS)

    Sommer, C. M.; Lemm, G.; Hohenstein, E.; Bellemann, N.; Stampfl, U.; Goezen, A. S.; Rassweiler, J.; Kauczor, H. U.; Radeleff, B. A.; Pereira, P. L.

    2013-01-01

    Purpose. This study was designed to evaluate the clinical efficacy of CT-guided bipolar and multipolar radiofrequency ablation (RF ablation) of renal cell carcinoma (RCC) and to analyze specific technical aspects between both technologies. Methods. We included 22 consecutive patients (3 women; age 74.2 ± 8.6 years) after 28 CT-guided bipolar or multipolar RF ablations of 28 RCCs (diameter 2.5 ± 0.8 cm). Procedures were performed with a commercially available RF system (Celon AG Olympus, Berlin, Germany). Technical aspects of RF ablation procedures (ablation mode [bipolar or multipolar], number of applicators and ablation cycles, overall ablation time and deployed energy, and technical success rate) were analyzed. Clinical results (local recurrence-free survival and local tumor control rate, renal function [glomerular filtration rate (GFR)]) and complication rates were evaluated. Results. Bipolar RF ablation was performed in 12 procedures and multipolar RF ablation in 16 procedures (2 applicators in 14 procedures and 3 applicators in 2 procedures). One ablation cycle was performed in 15 procedures and two ablation cycles in 13 procedures. Overall ablation time and deployed energy were 35.0 ± 13.6 min and 43.7 ± 17.9 kJ. Technical success rate was 100 %. Major and minor complication rates were 4 and 14 %. At an imaging follow-up of 15.2 ± 8.8 months, local recurrence-free survival was 14.4 ± 8.8 months and local tumor control rate was 93 %. GFR did not deteriorate after RF ablation (50.8 ± 16.6 ml/min/1.73 m 2 before RF ablation vs. 47.2 ± 11.9 ml/min/1.73 m 2 after RF ablation; not significant). Conclusions. CT-guided bipolar and multipolar RF ablation of RCC has a high rate of clinical success and low complication rates. At short-term follow-up, clinical efficacy is high without deterioration of the renal function.

  17. CT-guided Bipolar and Multipolar Radiofrequency Ablation (RF Ablation) of Renal Cell Carcinoma: Specific Technical Aspects and Clinical Results

    Energy Technology Data Exchange (ETDEWEB)

    Sommer, C. M., E-mail: christof.sommer@med.uni-heidelberg.de [University Hospital Heidelberg, INF 110, Department of Diagnostic and Interventional Radiology (Germany); Lemm, G.; Hohenstein, E. [Minimally Invasive Therapies and Nuclear Medicine, SLK Kliniken Heilbronn GmbH, Clinic for Radiology (Germany); Bellemann, N.; Stampfl, U. [University Hospital Heidelberg, INF 110, Department of Diagnostic and Interventional Radiology (Germany); Goezen, A. S.; Rassweiler, J. [Clinic for Urology, SLK Kliniken Heilbronn GmbH (Germany); Kauczor, H. U.; Radeleff, B. A. [University Hospital Heidelberg, INF 110, Department of Diagnostic and Interventional Radiology (Germany); Pereira, P. L. [Minimally Invasive Therapies and Nuclear Medicine, SLK Kliniken Heilbronn GmbH, Clinic for Radiology (Germany)

    2013-06-15

    Purpose. This study was designed to evaluate the clinical efficacy of CT-guided bipolar and multipolar radiofrequency ablation (RF ablation) of renal cell carcinoma (RCC) and to analyze specific technical aspects between both technologies. Methods. We included 22 consecutive patients (3 women; age 74.2 {+-} 8.6 years) after 28 CT-guided bipolar or multipolar RF ablations of 28 RCCs (diameter 2.5 {+-} 0.8 cm). Procedures were performed with a commercially available RF system (Celon AG Olympus, Berlin, Germany). Technical aspects of RF ablation procedures (ablation mode [bipolar or multipolar], number of applicators and ablation cycles, overall ablation time and deployed energy, and technical success rate) were analyzed. Clinical results (local recurrence-free survival and local tumor control rate, renal function [glomerular filtration rate (GFR)]) and complication rates were evaluated. Results. Bipolar RF ablation was performed in 12 procedures and multipolar RF ablation in 16 procedures (2 applicators in 14 procedures and 3 applicators in 2 procedures). One ablation cycle was performed in 15 procedures and two ablation cycles in 13 procedures. Overall ablation time and deployed energy were 35.0 {+-} 13.6 min and 43.7 {+-} 17.9 kJ. Technical success rate was 100 %. Major and minor complication rates were 4 and 14 %. At an imaging follow-up of 15.2 {+-} 8.8 months, local recurrence-free survival was 14.4 {+-} 8.8 months and local tumor control rate was 93 %. GFR did not deteriorate after RF ablation (50.8 {+-} 16.6 ml/min/1.73 m{sup 2} before RF ablation vs. 47.2 {+-} 11.9 ml/min/1.73 m{sup 2} after RF ablation; not significant). Conclusions. CT-guided bipolar and multipolar RF ablation of RCC has a high rate of clinical success and low complication rates. At short-term follow-up, clinical efficacy is high without deterioration of the renal function.

  18. Microsecond enamel ablation with 10.6μm CO2 laser radiation

    Science.gov (United States)

    Góra, W. S.; McDonald, A.; Hand, D. P.; Shephard, J. D.

    2016-02-01

    Lasers have been previously been used for dental applications, however there remain issues with thermally-induced cracking. In this paper we investigate the impact of pulse length on CO2 laser ablation of human dental enamel. Experiments were carried in vitro on molar teeth without any modification to the enamel surface, such as grinding or polishing. In addition to varying the pulse length, we also varied pulse energy and focal position, to determine the most efficient ablation of dental hard tissue and more importantly to minimize or eradicate cracking. The maximum temperature rise during the multi pulse ablation process was monitored using a set of thermocouples embedded into the pulpal chamber. The application of a laser device in dental surgery allows removal of tissue with higher precision, which results in minimal loss of healthy dental tissue. In this study we use an RF discharge excited CO2 laser operating at 10.6μm. The wavelength of 10.6 μm overlaps with a phosphate band (PO3-4) absorption in dental hard tissue hence the CO2 laser radiation has been selected as a potential source for modification of the tissue. This research describes an in-depth analysis of single pulse laser ablation. To determine the parameters that are best suited for the ablation of hard dental tissue without thermal cracking, a range of pulse lengths (10-200 μs), and fluences (0-100 J/cm2) are tested. In addition, different laser focusing approaches are investigated to select the most beneficial way of delivering laser radiation to the surface (divergent/convergent beam). To ensure that these processes do not increase the temperature above the critical threshold and cause the necrosis of the tissue a set of thermocouples was placed into the pulpal chambers. Intermittent laser radiation was investigated with and without application of a water spray to cool down the ablation site and the adjacent area. Results show that the temperature can be kept below the critical threshold

  19. Theoretical analyses of the refractive implications of transepithelial PRK ablations.

    Science.gov (United States)

    Arba Mosquera, Samuel; Awwad, Shady T

    2013-07-01

    To analyse the refractive implications of single-step, transepithelial photorefractive keratectomy (TransPRK) ablations. A simulation for quantifying the refractive implications of TransPRK ablations has been developed. The simulation includes a simple modelling of corneal epithelial profiles, epithelial ablation profiles as well as refractive ablation profiles, and allows the analytical quantification of the refractive implications of TransPRK in terms of wasted tissue, achieved optical zone (OZ) and induced refractive error. Wasted tissue occurs whenever the actual corneal epithelial profile is thinner than the applied epithelial ablation profile, achieved OZ is reduced whenever the actual corneal epithelial profile is thicker than the applied epithelial ablation profile and additional refractive errors are induced whenever the actual difference centre-to-periphery in the corneal epithelial profile deviates from the difference in the applied epithelial ablation profile. The refractive implications of TransPRK ablations can be quantified using simple theoretical simulations. These implications can be wasted tissue (∼14 µm, if the corneal epithelial profile is thinner than the ablated one), reduced OZ (if the corneal epithelial profile is thicker than ablated one, very severe for low corrections) and additional refractive errors (∼0.66 D, if the centre-to-periphery progression of the corneal epithelial profile deviates from the progression of the ablated one). When TransPRK profiles are applied to normal, not previously treated, non-pathologic corneas, no specific refractive implications associated to the transepithelial profile can be anticipated; TransPRK would provide refractive outcomes equal to those of standard PRK. Adjustments for the planned OZ and, in the event of retreatments, for the target sphere can be easily derived.

  20. Precise femtosecond laser ablation of dental hard tissue: preliminary investigation on adequate laser parameters

    International Nuclear Information System (INIS)

    Hikov, Todor; Pecheva, Emilia; Petrov, Todor; Montgomery, Paul; Antoni, Frederic; Leong-Hoi, Audrey

    2017-01-01

    This work aims at evaluating the possibility of introducing state-of-the-art commercial femtosecond laser system in restorative dentistry by maintaining well-known benefits of lasers for caries removal, but also in overcoming disadvantages such as thermal damage of irradiated substrate. Femtosecond ablation of dental hard tissue is investigated by changing the irradiation parameters (pulsed laser energy, scanning speed and pulse repetition rate), assessed for enamel and dentin. The femtosecond laser system used in this work may be suitable for cavity preparation in dentin and enamel, due to the expected effective ablation and low temperature increase when using ultra short laser pulses. If adequate laser parameters are selected, this system seems to be promising for promoting a laser-assisted, minimally invasive approach in restorative dentistry. (paper)

  1. Electroporation ablation: A new energy modality for ablation of arrhythmogenic cardiac substrate

    NARCIS (Netherlands)

    van Driel, VJHM

    2016-01-01

    At the very end of the Direct Current (DC) era, low-energy DC ablation was demonstrated to cause myocardial lesions by non-thermal irreversible electroporation (IRE) (permanent formation of pores in the cell membrane, leading to cell death), without arcing and/or barotrauma. To eliminate rather

  2. Similarities and differences in ablative and non-ablative iron oxide nanoparticle hyperthermia cancer treatment

    Science.gov (United States)

    Petryk, Alicia A.; Misra, Adwiteeya; Kastner, Elliot J.; Mazur, Courtney M.; Petryk, James D.; Hoopes, P. Jack

    2015-03-01

    The use of hyperthermia to treat cancer is well studied and has utilized numerous delivery techniques, including microwaves, radio frequency, focused ultrasound, induction heating, infrared radiation, warmed perfusion liquids (combined with chemotherapy), and recently, metallic nanoparticles (NP) activated by near infrared radiation (NIR) and alternating magnetic field (AMF) based platforms. It has been demonstrated by many research groups that ablative temperatures and cytotoxicity can be produced with locally NP-based hyperthermia. Such ablative NP techniques have demonstrated the potential for success. Much attention has also been given to the fact that NP may be administered systemically, resulting in a broader cancer therapy approach, a lower level of tumor NP content and a different type of NP cancer therapy (most likely in the adjuvant setting). To use NP based hyperthermia successfully as a cancer treatment, the technique and its goal must be understood and utilized in the appropriate clinical context. The parameters include, but are not limited to, NP access to the tumor (large vs. small quantity), cancer cell-specific targeting, drug carrying capacity, potential as an ionizing radiation sensitizer, and the material properties (magnetic characteristics, size and charge). In addition to their potential for cytotoxicity, the material properties of the NP must also be optimized for imaging, detection and direction. In this paper we will discuss the differences between, and potential applications for, ablative and non-ablative magnetic nanoparticle hyperthermia.

  3. Multifunctional Cationic Lipid-Based Nanoparticles Facilitate Endosomal Escape and Reduction-Triggered Cytosolic siRNA Release

    Science.gov (United States)

    Gujrati, Maneesh; Malamas, Anthony; Shin, Tesia; Jin, Erlei; Sun, Lulu; Lu, Zheng-Rong

    2015-01-01

    Small interfering RNA (siRNA) has garnered much attention in recent years as a promising avenue for cancer gene therapy due to its ability to silence disease-related genes. Effective gene silencing is contingent upon the delivery of siRNA into the cytosol of target cells and requires the implementation of delivery systems possessing multiple functionalities to overcome delivery barriers. The present work explores the multifunctional properties and biological activity of a recently developed cationic lipid carrier, (1-aminoethyl)iminobis[N-(oleicylcysteinyl-1-amino-ethyl)propionamide]) (ECO). The physicochemical properties and biological activity of ECO/siRNA nanoparticles were assessed over a range of N/P ratios to optimize the formulation. Potent and sustained luciferase silencing in a U87 glioblastoma cell line was observed, even in the presence of serum proteins. ECO/siRNA nanoparticles exhibited pH-dependent membrane disruption at pH levels corresponding to various stages of the intracellular trafficking pathway. It was found that disulfide linkages created during nanoparticle formation enhanced the protection of siRNA from degradation and facilitated site-specific siRNA release in the cytosol by glutathione-mediated reduction. Confocal microscopy confirmed that ECO/siRNA nanoparticles readily escaped from late endosomes prior to cytosolic release of the siRNA cargo. These results demonstrate that the rationally designed multifunctionality of ECO/siRNA nanoparticles is critical for intracellular siRNA delivery and the continuing development of safe and effective delivery systems. PMID:25020033

  4. Photochemical internalisation, a minimally invasive strategy for light-controlled endosomal escape of cancer stem cell-targeting therapeutics.

    Science.gov (United States)

    Selbo, Pål Kristian; Bostad, Monica; Olsen, Cathrine Elisabeth; Edwards, Victoria Tudor; Høgset, Anders; Weyergang, Anette; Berg, Kristian

    2015-08-01

    Despite progress in radio-, chemo- and photodynamic-therapy (PDT) of cancer, treatment resistance still remains a major problem for patients with aggressive tumours. Cancer stem cells (CSCs) or tumour-initiating cells are intrinsically and notoriously resistant to conventional cancer therapies and are proposed to be responsible for the recurrence of tumours after therapy. According to the CSC hypothesis, it is imperative to develop novel anticancer agents or therapeutic strategies that take into account the biology and role of CSCs. The present review outlines our recent study on photochemical internalisation (PCI) using the clinically relevant photosensitiser TPCS2a/Amphinex® as a rational, non-invasive strategy for the light-controlled endosomal escape of CSC-targeting drugs. PCI is an intracellular drug delivery method based on light-induced ROS-generation and a subsequent membrane-disruption of endocytic vesicles, leading to cytosolic release of the entrapped drugs of interest. In different proof-of-concept studies we have demonstrated that PCI of CSC-directed immunotoxins targeting CD133, CD44, CSPG4 and EpCAM is a highly specific and effective strategy for killing cancer cells and CSCs. CSCs overexpressing CD133 are PDT-resistant; however, this is circumvented by PCI of CD133-targeting immunotoxins. In view of the fact that TPCS2a is not a substrate of the efflux pumps ABCG2 and P-glycoprotein (ABCB1), the PCI-method is a promising anti-CSC therapeutic strategy. Due to a laser-controlled exposure, PCI of CSC-targeting drugs will be confined exclusively to the tumour tissue, suggesting that this drug delivery method has the potential to spare distant normal stem cells.

  5. Pannexin2 oligomers localize into endosomal vesicles in mammalian cells while Pannexin1 channels traffic to the plasma membrane

    Directory of Open Access Journals (Sweden)

    Daniela eBoassa

    2015-02-01

    Full Text Available Pannexin2 (Panx2 is the largest of three members of the pannexin proteins. Pannexins are topologically related to connexins and innexins, but serve different functional roles than forming gap junctions. We previously showed that pannexins form oligomeric channels but unlike connexins and innexins, they form only single membrane channels. High levels of Panx2 mRNA and protein in the Central Nervous System (CNS have been documented. Whereas Pannexin1 (Panx1 is fairly ubiquitous and Pannexin3 (Panx3 is found in skin and connective tissue, both are fully glycosylated, traffic to the plasma membrane and have functions correlated with extracellular ATP release. Here, we describe trafficking and subcellular localizations of exogenous Panx2 and Panx1 protein expression in MDCK, HeLa and HEK293T cells as well as endogenous Panx1 and Panx2 patterns in the CNS. Panx2 was found in intracellular localizations, was partially N-glycosylated, and localizations were non-overlapping with Panx1. Confocal images of hippocampal sections immunolabeled for the astrocytic protein GFAP, Panx1 and Panx2 demonstrated that the two isoforms, Panx1 and Panx2, localized at different subcellular compartments in both astrocytes and neurons. Using recombinant fusions of Panx2 with appended genetic tags developed for correlated light and electron microscopy and then expressed in different cell lines, we determined that Panx2 is localized in the membrane of intracellular vesicles and not in the endoplasmic reticulum as initially indicated by calnexin colocalization experiments. Dual immunofluorescence imaging with protein markers for specific vesicle compartments showed that Panx2 vesicles are early endosomal in origin. In electron tomographic volumes, cross-sections of these vesicles displayed fine structural details and close proximity to actin filaments. Thus, pannexins expressed at different subcellular compartments likely exert distinct functional roles, particularly in the

  6. A voltage-gated calcium channel regulates lysosomal fusion with endosomes and autophagosomes and is required for neuronal homeostasis.

    Directory of Open Access Journals (Sweden)

    Xuejun Tian

    2015-03-01

    Full Text Available Autophagy helps deliver sequestered intracellular cargo to lysosomes for proteolytic degradation and thereby maintains cellular homeostasis by preventing accumulation of toxic substances in cells. In a forward mosaic screen in Drosophila designed to identify genes required for neuronal function and maintenance, we identified multiple cacophony (cac mutant alleles. They exhibit an age-dependent accumulation of autophagic vacuoles (AVs in photoreceptor terminals and eventually a degeneration of the terminals and surrounding glia. cac encodes an α1 subunit of a Drosophila voltage-gated calcium channel (VGCC that is required for synaptic vesicle fusion with the plasma membrane and neurotransmitter release. Here, we show that cac mutant photoreceptor terminals accumulate AV-lysosomal fusion intermediates, suggesting that Cac is necessary for the fusion of AVs with lysosomes, a poorly defined process. Loss of another subunit of the VGCC, α2δ or straightjacket (stj, causes phenotypes very similar to those caused by the loss of cac, indicating that the VGCC is required for AV-lysosomal fusion. The role of VGCC in AV-lysosomal fusion is evolutionarily conserved, as the loss of the mouse homologues, Cacna1a and Cacna2d2, also leads to autophagic defects in mice. Moreover, we find that CACNA1A is localized to the lysosomes and that loss of lysosomal Cacna1a in cerebellar cultured neurons leads to a failure of lysosomes to fuse with endosomes and autophagosomes. Finally, we show that the lysosomal CACNA1A but not the plasma-membrane resident CACNA1A is required for lysosomal fusion. In summary, we present a model in which the VGCC plays a role in autophagy by regulating the fusion of AVs with lysosomes through its calcium channel activity and hence functions in maintaining neuronal homeostasis.

  7. High-Resolution Spectroscopy of Laser Ablation Plumes Using Laser-Induced Fluorescence

    Energy Technology Data Exchange (ETDEWEB)

    Harilal, Sivanandan S.; LaHaye, Nicole L.; Phillips, Mark C.

    2017-02-06

    We used a CW laser as a narrow-band (~50kHz) tunable LIF excitation source to probe absorption from selected atomic transitions (Al, U etc. ) in a ns laser ablation plume. A comparison of fluorescence signal with respect to emission spectroscopy show significant increase in the magnitude and persistence from selected Al and U transitions in a LIBS plume. The high spectral resolution provided by the LIF measurement allows peaks to be easily separated even if they overlap in the emission spectra.

  8. MRI-guided prostate focal laser ablation therapy using a mechatronic needle guidance system

    Science.gov (United States)

    Cepek, Jeremy; Lindner, Uri; Ghai, Sangeet; Davidson, Sean R. H.; Trachtenberg, John; Fenster, Aaron

    2014-03-01

    Focal therapy of localized prostate cancer is receiving increased attention due to its potential for providing effective cancer control in select patients with minimal treatment-related side effects. Magnetic resonance imaging (MRI)-guided focal laser ablation (FLA) therapy is an attractive modality for such an approach. In FLA therapy, accurate placement of laser fibers is critical to ensuring that the full target volume is ablated. In practice, error in needle placement is invariably present due to pre- to intra-procedure image registration error, needle deflection, prostate motion, and variability in interventionalist skill. In addition, some of these sources of error are difficult to control, since the available workspace and patient positions are restricted within a clinical MRI bore. In an attempt to take full advantage of the utility of intraprocedure MRI, while minimizing error in needle placement, we developed an MRI-compatible mechatronic system for guiding needles to the prostate for FLA therapy. The system has been used to place interstitial catheters for MRI-guided FLA therapy in eight subjects in an ongoing Phase I/II clinical trial. Data from these cases has provided quantification of the level of uncertainty in needle placement error. To relate needle placement error to clinical outcome, we developed a model for predicting the probability of achieving complete focal target ablation for a family of parameterized treatment plans. Results from this work have enabled the specification of evidence-based selection criteria for the maximum target size that can be confidently ablated using this technique, and quantify the benefit that may be gained with improvements in needle placement accuracy.

  9. Inertial effects in laser-driven ablation

    International Nuclear Information System (INIS)

    Harrach, R.J.; Szeoke, A.; Howard, W.M.

    1983-01-01

    The gasdynamic partial differential equations (PDE's) governing the motion of an ablatively accelerated target (rocket) contain an inertial force term that arises from acceleration of the reference frame in which the PDE's are written. We give a simple, intuitive description of this effect, and estimate its magnitude and parametric dependences by means of approximate analytical formulas inferred from our computer hydrocode calculations. Often this inertial term is negligible, but for problems in the areas of laser fusion and laser equation of state studies we find that it can substantially reduce the attainable hydrodynamic efficiency of acceleration and implosion

  10. Ablative Material Testing at Lewis Rocket Lab

    Science.gov (United States)

    1997-01-01

    The increasing demand for a low-cost, reliable way to launch commercial payloads to low- Earth orbit has led to the need for inexpensive, expendable propulsion systems for new launch vehicles. This, in turn, has renewed interest in less complex, uncooled rocket engines that have combustion chambers and exhaust nozzles fabricated from ablative materials. A number of aerospace propulsion system manufacturers have utilized NASA Lewis Research Center's test facilities with a high degree of success to evaluate candidate materials for application to new propulsion devices.

  11. Laser systems for ablative fractional resurfacing

    DEFF Research Database (Denmark)

    Paasch, Uwe; Haedersdal, Merete

    2011-01-01

    of a variety of skin conditions, primarily chronically photodamaged skin, but also acne and burn scars. In addition, it is anticipated that AFR can be utilized in the laser-assisted delivery of topical drugs. Clinical efficacy coupled with minimal downtime has driven the development of various fractional...... ablative laser systems. Fractionated CO(2) (10,600-nm), erbium yttrium aluminum garnet, 2940-nm and yttrium scandium gallium garnet, 2790-nm lasers are available. In this article, we present an overview of AFR technology, devices and histopathology, and we summarize the current clinical possibilities...

  12. Resonant laser ablation: mechanisms and applications

    International Nuclear Information System (INIS)

    Anderson, J.E.; Bodla, R.; Eiden, G.C.; Nogar, N.S.; Smith, C.H.

    1996-01-01

    Resonant laser ablation (RLA) typically relies on irradiation of a sample in a mass spectrometer with modest intensity laser pulses tuned to a one or two photon resonant transition in the analyte of interest. This paper shows that RLA is well suited for highly sensitive analyses of complex samples. The examples actually studied are trace components in rhenium and technetium in nickel. The authors also studied the 2+1 multiphoton ionization spectrum of iron-56 detected by RLA of Re containing 70 ppm iron. Two-photon transition rates for Fe transitions were calculated perturbatively and found to agree semi-quantitatively with experimentally observed intensities. 17 refs., 3 figs

  13. Obtention of Ti nanoparticles by laser ablation

    International Nuclear Information System (INIS)

    Diaz E, J.R.; Escobar A, L.; Camps, E.; Santiago, P.; Ascencio, J.

    2002-01-01

    The obtention of Ti nanoparticles around 5-30 nm diameter through the laser ablation technique is reported. The formation of nanoparticles is carried out in He atmosphere to different pressures, placing directly in Si substrates (100) and in Cu grids. The results show that the work pressure is an important parameter that allows to control the nanoparticles size. Also the plasma characterization results are presented where the Ti II is the predominant specie with an average kinetic energy of 1824 eV. (Author)

  14. Excimer laser ablation of the cornea

    Science.gov (United States)

    Pettit, George H.; Ediger, Marwood N.; Weiblinger, Richard P.

    1995-03-01

    Pulsed ultraviolet laser ablation is being extensively investigated clinically to reshape the optical surface of the eye and correct vision defects. Current knowledge of the laser/tissue interaction and the present state of the clinical evaluation are reviewed. In addition, the principal findings of internal Food and Drug Administration research are described in some detail, including a risk assessment of the laser-induced-fluorescence and measurement of the nonlinear optical properties of cornea during the intense UV irradiation. Finally, a survey is presented of the alternative laser technologies being explored for this ophthalmic application.

  15. Laser systems for ablative fractional resurfacing

    DEFF Research Database (Denmark)

    Paasch, Uwe; Haedersdal, Merete

    2011-01-01

    ablative laser systems. Fractionated CO(2) (10,600-nm), erbium yttrium aluminum garnet, 2940-nm and yttrium scandium gallium garnet, 2790-nm lasers are available. In this article, we present an overview of AFR technology, devices and histopathology, and we summarize the current clinical possibilities...... of a variety of skin conditions, primarily chronically photodamaged skin, but also acne and burn scars. In addition, it is anticipated that AFR can be utilized in the laser-assisted delivery of topical drugs. Clinical efficacy coupled with minimal downtime has driven the development of various fractional...

  16. Ablation of polymers by ultraviolet pulsed laser

    International Nuclear Information System (INIS)

    Brezini, A.; Benharrats, N.

    1993-08-01

    The surface modifications of different polymers treated by far UV-Excimer laser (λ = 193mn, 248, 308nm) are analysed by X-Ray Photoelectrons Spectroscopy. The main feature observed depends strongly on the absorption coefficients. For the high absorbing polymers such (PVC, PS, PI,...) the mechanism of the UV-Excimer Laser interaction appears to be governed by an ablative photodecomposition process (APD) with an APD threshold. In the other limit, i.e. low absorbing polymer the interaction leads to a photothermal process. (author). 51 refs, 24 figs, 7 tabs

  17. Laser ablation studies in southern Africa

    Science.gov (United States)

    McKenzie, Edric; Forbes, A.; Turner, G. R.; Michaelis, Max M.

    2000-08-01

    With the launch of the South African National Laser Centre, new programs will need to be defined. Medical, environmental and industrial laser applications must obviously take top priority -- as opposed to the uranium isotope separation and military applications of the past. We argue however, that a small effort in laser ablation for space propulsion is justifiable, since a few very large CO2 lasers are available and since two tentative propulsion experiments have already been conducted in South Africa. We attempt to give LISP (Laser Impulse Space Propulsion) an equatorial and a Southern dimension.

  18. Rail gun performance and plasma characteristics due to wall ablation

    Science.gov (United States)

    Ray, P. K.

    1986-01-01

    The experiment of Bauer, et al. (1982) is analyzed by considering wall ablation and viscous drag in the plasma. Plasma characteristics are evaluated through a simple fluid-mechanical analysis considering only wall ablation. By equating the energy dissipated in the plasma with the radiation heat loss, the average properties of the plasma are determined as a function of time.

  19. Ganglion Plexus Ablation in Advanced Atrial Fibrillation: The AFACT Study

    NARCIS (Netherlands)

    Driessen, Antoine H. G.; Berger, Wouter R.; Krul, Sébastien P. J.; van den Berg, Nicoline W. E.; Neefs, Jolien; Piersma, Femke R.; Chan Pin Yin, Dean R. P. P.; de Jong, Jonas S. S. G.; van Boven, WimJan P.; de Groot, Joris R.

    2016-01-01

    Patients with long duration of atrial fibrillation (AF), enlarged atria, or failed catheter ablation have advanced AF and may require more extensive treatment than pulmonary vein isolation. The aim of this study was to investigate the efficacy and safety of additional ganglion plexus (GP) ablation

  20. Delayed Development of Pneumothorax After Pulmonary Radiofrequency Ablation

    International Nuclear Information System (INIS)

    Clasen, Stephan; Kettenbach, Joachim; Kosan, Bora; Aebert, Hermann; Schernthaner, Melanie; Kroeber, Stefan-Martin; Boemches, Andrea; Claussen, Claus D.; Pereira, Philippe L.

    2009-01-01

    Acute pneumothorax is a frequent complication after percutaneous pulmonary radiofrequency (RF) ablation. In this study we present three cases showing delayed development of pneumothorax after pulmonary RF ablation in 34 patients. Our purpose is to draw attention to this delayed complication and to propose a possible approach to avoid this major complication. These three cases occurred subsequent to 44 CT-guided pulmonary RF ablation procedures (6.8%) using either internally cooled or multitined expandable RF electrodes. In two patients, the pneumothorax, being initially absent at the end of the intervention, developed without symptoms. One of these patients required chest drain placement 32 h after RF ablation, and in the second patient therapy remained conservative. In the third patient, a slight pneumothorax at the end of the intervention gradually increased and led into tension pneumothorax 5 days after ablation procedure. Underlying bronchopleural fistula along the coagulated former electrode track was diagnosed in two patients. In conclusion, delayed development of pneumothorax after pulmonary RF ablation can occur and is probably due to underlying bronchopleural fistula, potentially leading to tension pneumothorax. Patients and interventionalists should be prepared for delayed onset of this complication, and extensive track ablation following pulmonary RF ablation should be avoided.

  1. Spectroscopic photoacoustic imaging of radiofrequency ablation in the left atrium

    NARCIS (Netherlands)

    S. Iskander-Rizk (Sophinese); P. Kruizinga (Pieter); A.F.W. van der Steen (Ton); G. van Soest (Gijs)

    2018-01-01

    textabstractCatheter-based radiofrequency ablation for atrial fibrillation has long-term success in 60-70% of cases. A better assessment of lesion quality, depth, and continuity could improve the procedure’s outcome. We investigate here photoacoustic contrast between ablated and healthy atrial-wall

  2. Catheter Ablation of Focal Atrial Tachycardia Using Remote Magnetic Navigation

    DEFF Research Database (Denmark)

    Liu, Xiao-Yu; Jacobsen, Peter Karl; Pehrson, Steen

    2018-01-01

    , a total of 56 atrial foci were found. Acute success of the primary ablation was obtained in 52 patients (98%). Mean procedure duration was 109 ± 35 min, ablation duration was 401 sec (interquartile range [IQR], 332 sec), and fluoroscopy time was 5.0 min (IQR, 3.0 min). After a mean follow-up of 31 ± 18...

  3. Subtotal ablation of parietal epithelial cells induces crescent formation.

    NARCIS (Netherlands)

    Sicking, E.M.; Fuss, A.; Uhlig, S.; Jirak, P.; Dijkman, H.; Wetzels, J.; Engel, D.R.; Urzynicok, T.; Heidenreich, S.; Kriz, W.; Kurts, C.; Ostendorf, T.; Floege, J.; Smeets, B.; Moeller, M.J.

    2012-01-01

    Parietal epithelial cells (PECs) of the renal glomerulus contribute to the formation of both cellular crescents in rapidly progressive GN and sclerotic lesions in FSGS. Subtotal transgenic ablation of podocytes induces FSGS but the effect of specific ablation of PECs is unknown. Here, we established

  4. Unusual tumour ablations: report of difficult and interesting cases

    OpenAIRE

    Mauri, Giovanni; Nicosia, Luca; Varano, Gianluca Maria; Shyn, Paul; Sartori, Sergio; Tombesi, Paola; Di Vece, Francesca; Orsi, Franco; Solbiati, Luigi

    2017-01-01

    Image-guided ablations are nowadays applied in the treatment of a wide group of diseases and in different organs and regions, and every day interventional radiologists have to face more difficult and unusual cases of tumour ablation. In the present case review, we report four difficult and unusual cases, reporting some tips and tricks for a successful image-guided treatment.

  5. Low vulnerability of the right phrenic nerve to electroporation ablation

    NARCIS (Netherlands)

    van Driel, Vincent J. H. M.; Neven, KGEJ; van Wessel, Harri; Vink, Aryan; Doevendans, Pieter A. F. M.; Wittkampf, Fred H. M.

    BACKGROUND Circular electroporation ablation is a novel ablation modality for electrical pulmonary vein isolation. With a single 200-3 application, deep circular myocardial lesions can be created. However, the acute and chronic effects of this energy source on phrenic nerve (PN) function are

  6. Pulsed Tm:YAG laser ablation of knee joint tissues

    Science.gov (United States)

    Shi, Wei-Qiang; Vari, Sandor G.; Duffy, J. T.; Miller, J. M.; Weiss, Andrew B.; Fishbein, Michael C.; Grundfest, Warren S.

    1992-06-01

    We investigated the effect of a free-running 2.01 micron pulsed Tm:YAG laser on bovine knee joint tissues. Ablation rates of fresh fibrocartilage, hyaline cartilage, and bone were measured in saline as a function of laser fluence (160 - 640 J/cm2) and fiber core size (400 and 600 microns). All tissues could be effectively ablated and the ablation rate increased linearly with the increasing fluence. Use of fibers of different core sizes, while maintaining constant energy fluence, did not result in significant difference in ablation rate. Histology analyses of the ablated tissue samples reveal average Tm:YAG radiation induced thermal damage (denatunalization) zones ranging between 130 and 540 microns, depending on the laser parameters and the tissue type.

  7. Nanoparticle fabrication of hydroxyapatite by laser ablation in water

    International Nuclear Information System (INIS)

    Musaev, O. R.; Wieliczka, D. M.; Wrobel, J. M.; Kruger, M. B.; Dusevich, V.

    2008-01-01

    Synthetic polycrystalline hydroxyapatite was ablated in water with 337 nm radiation from a UV nitrogen pulsed laser. According to transmission electron microscopy micrographs, the ablated particles were approximately spherical and had a size of ∼80 nm. Raman spectroscopic analysis demonstrated that particles had the same structure as the original crystal. X-ray photoelectron spectroscopy showed that the surface chemical composition was close to that of the original material. The characteristics of the ablated particles and estimations of the temperature rise of the hydroxyapatite surface under laser irradiation are consistent with the mechanism of explosive boiling being responsible for ablation. The experimental observations offer the basis for preparation of hydroxyapatite nanoparticles by laser ablation in water

  8. Chemothermal Therapy for Localized Heating and Ablation of Tumor

    Directory of Open Access Journals (Sweden)

    Zhong-Shan Deng

    2013-01-01

    Full Text Available Chemothermal therapy is a new hyperthermia treatment on tumor using heat released from exothermic chemical reaction between the injected reactants and the diseased tissues. With the highly minimally invasive feature and localized heating performance, this method is expected to overcome the ubiquitous shortcomings encountered by many existing hyperthermia approaches in ablating irregular tumor. This review provides a relatively comprehensive review on the latest advancements and state of the art in chemothermal therapy. The basic principles and features of two typical chemothermal ablation strategies (acid-base neutralization-reaction-enabled thermal ablation and alkali-metal-enabled thermal/chemical ablation are illustrated. The prospects and possible challenges facing chemothermal ablation are analyzed. The chemothermal therapy is expected to open many clinical possibilities for precise tumor treatment in a minimally invasive way.

  9. Computational modeling of ultra-short-pulse ablation of enamel

    Energy Technology Data Exchange (ETDEWEB)

    London, R.A.; Bailey, D.S.; Young, D.A. [and others

    1996-02-29

    A computational model for the ablation of tooth enamel by ultra-short laser pulses is presented. The role of simulations using this model in designing and understanding laser drilling systems is discussed. Pulses of duration 300 sec and intensity greater than 10{sup 12} W/cm{sup 2} are considered. Laser absorption proceeds via multi-photon initiated plasma mechanism. The hydrodynamic response is calculated with a finite difference method, using an equation of state constructed from thermodynamic functions including electronic, ion motion, and chemical binding terms. Results for the ablation efficiency are presented. An analytic model describing the ablation threshold and ablation depth is presented. Thermal coupling to the remaining tissue and long-time thermal conduction are calculated. Simulation results are compared to experimental measurements of the ablation efficiency. Desired improvements in the model are presented.

  10. Picosecond laser ablation of porcine sclera

    Science.gov (United States)

    Góra, Wojciech S.; Harvey, Eleanor M.; Dhillon, Baljean; Parson, Simon H.; Maier, Robert R. J.; Hand, Duncan P.; Shephard, Jonathan D.

    2013-03-01

    Lasers have been shown to be successful in certain medical procedures and they have been identified as potentially making a major contribution to the development of minimally invasive procedures. However, the uptake is not as widespread and there is scope for many other applications where laser devices may offer a significant advantage in comparison to the traditional surgical tools. The purpose of this research is to assess the potential of using a picosecond laser for minimally invasive laser sclerostomy. Experiments were carried out on porcine scleral samples due to the comparable properties to human tissue. Samples were prepared with a 5mm diameter trephine and were stored in lactated Ringer's solution. After laser machining, the samples were fixed in 3% glutaraldehyde, then dried and investigated under SEM. The laser used in the experiments is an industrial picosecond TRUMPF TruMicro laser operating at a wavelength of 1030nm, pulse length of 6ps, repetition rate of 1 kHz and a focused spot diameter of 30μm. The laser beam was scanned across the samples with the use of a galvanometer scan head and various ablation patterns were investigated. Processing parameters (pulse energy, spot and line separation) which allow for the most efficient laser ablation of scleral tissue without introducing any collateral damage were investigated. The potential to create various shapes, such as linear incisions, square cavities and circular cavities was demonstrated.

  11. Recolonization of laser-ablated bacterial biofilm.

    Science.gov (United States)

    Nandakumar, Kanavillil; Obika, Hideki; Utsumi, Akihiro; Toshihiko, Ooie; Yano, Tetsuo

    2004-01-20

    The recolonization of laser-ablated bacterial monoculture biofilm was studied in the laboratory by using a flow-cytometer system. The marine biofilm-forming bacterium Pseudoalteromonas carrageenovora was used to develop biofilms on titanium coupons. Upon exposure to a low-power pulsed irradiation from an Nd:YAG laser, the coupons with biofilm were significantly reduced both in terms of total viable count (TVC) and area cover. The energy density used for a pulse of 5 ns was 0.1 J/cm(2) and the durations of irradiation exposure were 5 and 10 min. When placed in a flow of dilute ZoBell marine broth medium (10%) the laser-destructed bacterial film in a flow-cytometer showed significant recovery over a period of time. The flow of medium was regulated at 3.2 ml/min. The increase in area cover and TVC, however, was significantly less than that observed for nonirradiated control (t-test, Precolonization compared to control was thought be due to the lethal and sublethal impacts of laser irradiation on bacteria. This observation thus provided data on the online recolonization speed of biofilm, which is important when considering pulsed laser irradiation as an ablating technique of biofilm formation and removal in natural systems. Copyright 2003 Wiley Periodicals, Inc.

  12. Radiofrequency ablation for hepatocellular carcinoma: assistant techniques for difficult cases.

    Science.gov (United States)

    Inoue, Tatsuo; Minami, Yasunori; Chung, Hobyung; Hayaishi, Sousuke; Ueda, Taisuke; Tatsumi, Chie; Takita, Masahiro; Kitai, Satoshi; Hatanaka, Kinuyo; Ishikawa, Emi; Yada, Norihisa; Hagiwara, Satoru; Ueshima, Kazuomi; Kudo, Masatoshi

    2010-07-01

    To confirm the safety and effectiveness of techniques to assist radiofrequency ablation (RFA) for difficult cases, we retrospectively evaluated successful treatment rates, early complications and local tumor progressions. Between June 1999 and April 2009, a total of 341 patients with 535 nodules were treated as difficult cases. Artificial pleural effusion assisted ablation was performed on 64 patients with 82 nodules. Artificial ascites-assisted ablation was performed on 11 patients with 13 nodules. Cooling by endoscopic nasobiliary drainage (ENBD) tube-assisted ablation was performed on 6 patients with 8 nodules. When the tumors were not well visualized with conventional B-mode ultrasonography (US), contrast-enhanced US-assisted ablation with Levovist or Sonazoid or virtual CT sonography-assisted ablation was performed. Contrast-enhanced US-assisted ablation was performed on 139 patients with 224 nodules and virtual CT sonography-assisted ablation was performed on 121 patients with 209 nodules. In total, complete ablation was achieved in 514 of 535 (96%) nodules in difficult cases. For RFA with artificial pleural effusion, artificial ascites and ENBD, complete response was confirmed in all cases. For contrast-enhanced US- and CT sonography-assisted ablation, complete response was 95%. Early complications were recognized in 24 cases (4.5%). All cases recovered with no invasive treatment. Local tumor recurrence was investigated in 377 nodules of 245 patients, and 69 (18%) nodules were positive. Tumor recurrences in each assisted technique were 14.7% in artificial pleural effusion cases, 7% in artificial ascites, 12.5% in ENBD tube cases, 31% in virtual CT sonography, and 8.5% in contrast-enhanced US. Although local tumor progression needs to be carefully monitored, assisted techniques of RFA for difficult cases are well tolerated and expand the indications of RFA. Copyright (c) 2010 S. Karger AG, Basel.

  13. The Success Rate of Initial {sup 131I} Ablation in Differentiated Thyroid Cancer: Comparison Between Less strict and Very Strict Low Iodine Diets

    Energy Technology Data Exchange (ETDEWEB)

    Yoo, Ik Dong; Kim, Sung Hoon; Seo, Ye Young; Oh, Jin Kyoung; O, Joo Hyun; Chung, Soo Kyo [The Catholic Univ. of Korea, Seoul (Korea, Republic of)

    2012-03-15

    To decrease the risk of recurrence or metastasis in differentiated thyroid cancer (DTC), selected patients receive radioactive iodine ablation of remnant thyroid tissue or tumor. A low iodine diet can enhance uptake of radioactive iodine. We compared the success rates of radioactive iodine ablation therapy in patients who followed two different low iodine diets (LIDs). The success rates of postsurgical radioactive iodine ablation in DTC patients receiving empiric doses of 150 mCi were retrospectively reviewed. First-time radioactive iodine ablation therapy was done in 71 patients following less strict LID. Less strict LID restricted seafood, iodized salt, egg yolk, dairy products, processed meat, instant prepared meals, and multivitamins. Very strict LID additionally restricted rice, freshwater fish, spinach, and soybean products. Radioactive iodine ablation therapy was considered successful when follow up {sup 123I} whole body scan was negative and stimulated serum thyroglobulin level was less than 2.0 ng/mL. The success rate of patients following less strict LID was 80.3% and for very strict LID 75.6%. There was no statistically significant difference in the success rates between the two LID groups (P=0.48). Very strict LID may not contribute to improving the success rate of initial radioactive iodine ablation therapy at the cost of great inconvenience to the patient.

  14. Ventricular fibrillation occurring after atrioventricular node ablation despite minimal difference between pre- and post-ablation heart rates.

    Science.gov (United States)

    Squara, F; Theodore, G; Scarlatti, D; Ferrari, E

    2017-02-01

    We report the case of an 82-year-old man presenting with ventricular fibrillation (VF) occurring acutely after atrioventricular node (AVN) ablation. This patient had severe valvular cardiomyopathy, chronic atrial fibrillation (AF), and underwent prior to the AVN ablation a biventricular implantable cardiac defibrillator positioning. The VF was successfully cardioverted with one external electrical shock. What makes this presentation original is that the pre-ablation spontaneous heart rate in AF was slow (84 bpm), and that VF occurred after ablation despite a minimal heart rate drop of only 14 bpm. VF is the most feared complication of AVN ablation, but it had previously only been described in case of acute heart rate drop after ablation of at least 30 bpm (and more frequently>50 bpm). This case report highlights the fact that VF may occur after AVN ablation regardless of the heart rate drop, rendering temporary fast ventricular pacing mandatory whatever the pre-ablation heart rate. Copyright © 2016 Elsevier Masson SAS. All rights reserved.

  15. Improvement of the surface finish obtained by laser ablation with a Nd: YAG laser on pre-ablated tool steel

    CSIR Research Space (South Africa)

    Steyn, J

    2007-01-01

    Full Text Available . In recent years, these lasers have been used in other fields, such as laser ablation of small tools for plastics injection moulding. Laser ablation is a technology that is investigated as a method to improve the surface finish in tool steel. Different...

  16. Clinical effects of non-ablative and ablative fractional lasers on various hair disorders: a case series of 17 patients.

    Science.gov (United States)

    Cho, Suhyun; Choi, Min Ju; Zheng, Zhenlong; Goo, Boncheol; Kim, Do-Young; Cho, Sung Bin

    2013-04-01

    Both ablative and non-ablative fractional lasers have been applied to various uncommon hair disorders. The purpose of this study was to demonstrate the clinical effects of fractional laser therapy on the course of primary follicular and perifollicular pathologies and subsequent hair regrowth. A retrospective review of 17 patients with uncommon hair disorders - including ophiasis, autosomal recessive woolly hair/hypotrichosis, various secondary cicatricial alopecias, pubic hypotrichosis, frontal fibrosing alopecia, and perifolliculitis abscedens et suffodiens - was conducted. All patients had been treated with non-ablative and/or ablative fractional laser therapies. The mean clinical improvement score in these 17 patients was 2.2, while the mean patient satisfaction score was 2.5. Of the 17 subjects, 12 (70.6%) demonstrated a clinical response to non-ablative and/or ablative fractional laser treatments, including individuals with ophiasis, autosomal recessive woolly hair/hypotrichosis, secondary cicatricial alopecia (scleroderma and pressure-induced alopecia), frontal fibrosing alopecia, and perifolliculitis abscedens et suffodiens. Conversely, patients with long-standing ophiasis, surgical scar-induced secondary cicatricial alopecia, and pubic hypotrichosis did not respond to fractional laser therapy. Our findings demonstrate that the use of non-ablative and/or ablative fractional lasers promoted hair growth in certain cases of uncommon hair disorders without any remarkable side effects.

  17. Transcatheter radiofrequency ablation under the guidance of three-dimensional mapping for the treatment of complex cardiac arrhythmias

    International Nuclear Information System (INIS)

    Hong Lang; Wang Hong; Lai Hengli; Ying Qiulin; Chen Zhangqiang; Lu Linxiang; Qiu Yun; Xiao Chengwei

    2010-01-01

    Objective: To investigate the effectiveness and safety of transcatheter radiofrequency ablation guided by a three-dimensional mapping system (Ensite or Carto) for the treatment of complex cardiac arrhythmias. Methods: A cohort of 123 consecutive hospitalized inpatients during the period from February 2006 to December 2008 were selected for this study. These patients suffered from various arrhythmias, including paroxysmal atrial fibrillation (n = 58), persistent or permanent atrial fibrillation (n = 10), atrial flutter (n = 13), atrial tachycardia (n = 12) and ventricular tachycardia or frequent ventricular premature beats (n = 30). Transcatheter radiofrequency ablation for arrhythmias was performed under the guidance of an EnSite3000 / NavX or Array mapping system in 80 cases, and under the guidance of a CARTO mapping system in the remaining 43 cases. Results: Successful ablation of arrhythmias was obtained by single operation in 106 cases (86.18%), including 59 cases with atrial fibrillation, 11 cases with atrial flutter, 10 cases with atrial tachycardia, and 26 cases with ventricular tachycardia or premature ventricular beat.Ablation procedure was carried out and was successful in 10 cases with a successful rate of 94.31%, including 5 cases with atrial fibrillation, 1 case with recurred atrial flutter, 1 case with recurrent atrial tachycardia, and 3 cases with ventricular tachycardia or premature ventricular beat.After operation, complications occurred in 6 cases, including cardiac tamponade in 4 cases, distal embolism of the left anterior descending coronary artery in 1 case, and pulmonary embolism in 1 case. Conclusion: Three-dimensional mapping system can clearly and stereoscopically display the cardiac structures. Therefore, this technique is of great value in guiding the transcatheter radiofrequency ablation for complex arrhythmias, in improving the success rate of ablation and in increasing the safety of the procedure. (authors)

  18. Time-resolved photoluminescence for evaluating laser-induced damage during dielectric stack ablation in silicon solar cells

    Energy Technology Data Exchange (ETDEWEB)

    Parola, Stéphanie [Université de Lyon, Institut des Nanotechnologies de Lyon INL-UMR5270, CNRS, INSA Lyon, Villeurbanne, F-69621 (France); Blanc-Pélissier, Danièle, E-mail: daniele.blanc@insa-lyon.fr [Université de Lyon, Institut des Nanotechnologies de Lyon INL-UMR5270, CNRS, INSA Lyon, Villeurbanne, F-69621 (France); Barbos, Corina; Le Coz, Marine [Université de Lyon, Institut des Nanotechnologies de Lyon INL-UMR5270, CNRS, INSA Lyon, Villeurbanne, F-69621 (France); Poulain, Gilles [TOTAL MS—New Energies, R& D Division, La Défense (France); Lemiti, Mustapha [Université de Lyon, Institut des Nanotechnologies de Lyon INL-UMR5270, CNRS, INSA Lyon, Villeurbanne, F-69621 (France)

    2016-06-30

    Highlights: • Ablation of Al{sub 2}O{sub 3} and Al{sub 2}O{sub 3}/SiN{sub x} on Si substrates was performed with a nanosecond UV laser. • Ablation thresholds were found in good agreement with COMSOL simulation, around 0.85 and 0.95 J cm{sup −2} for Al{sub 2}O{sub 3} and Al{sub 2}O{sub 3}/SiN{sub X}, respectively. • Laser-induced damage was evaluated at room temperature by time-resolved photoluminescence decay with a single photon counting detector. • Minority carrier lifetime in silicon as a function of the ablation fluence was derived from the photoluminescence decay and related to the thickness of the heat affected zone. • Quantitative measurements of laser-induced damage can be used to evaluate laser ablation of dielectrics in photovoltaics. - Abstract: Selective laser ablation of dielectric layers on crystalline silicon wafers was investigated for solar cell fabrication. Laser processing was performed on Al{sub 2}O{sub 3}, and bi-layers Al{sub 2}O{sub 3}/SiN{sub X}:H with a nanosecond UV laser at various energy densities ranging from 0.4 to 2 J cm{sup −2}. Ablation threshold was correlated to the simulated temperature at the interface between the dielectric coatings and the silicon substrate. Laser-induced damage to the silicon substrate was evaluated by time-resolved photoluminescence. The minority carrier lifetime deduced from time-resolved photoluminescence was related to the depth of the heat affected zone in the substrate.

  19. Efficacy and Safety of Radiofrequency Ablation for Focal Hepatic Lesions Adjacent to Gallbladder: Reconfiguration of the Ablation Zone through Probe Relocation and Ablation Time Reduction.

    Science.gov (United States)

    Choi, In Young; Kim, Pyo Nyun; Lee, Sung Gu; Won, Hyung Jin; Shin, Yong Moon

    2017-10-01

    To evaluate the safety and efficacy of radiofrequency (RF) ablation for treatment of focal hepatic lesions adjacent to the gallbladder with electrode relocation and ablation time reduction. Thirty-nine patients who underwent RF ablation for focal hepatic lesions adjacent to the gallbladder (≤ 10 mm) were evaluated retrospectively from January 2011 to December 2014 (30 men and 9 women; age range, 51-85 y; mean age, 65 y). Of 36 patients with hepatocellular carcinoma, 3 had a second treatment for recurrence (mean tumor size, 15 mm ± 6). Patients were divided into 2 subgroups based on lesion distance from the gallbladder: nonabutting (> 5 mm; n = 19) and abutting (≤ 5 mm; n = 20). Electrodes were inserted parallel to the gallbladder through the center of a tumor in the nonabutting group and through the center of the expected ablation zone between a 5-mm safety zone on the liver side and the gallbladder in the abutting group. Ablation time was decreased in proportion to the transverse diameter of the expected ablation zone. Technical success and technical effectiveness rates were 89.7% and 97.4%, respectively, with no significant differences between groups (P = 1.00). Local tumor progression was observed in 3 patients (1 in the nonabutting group and 2 in the abutting group; P = 1.00). There were no major complications. The gallbladder was thickened in 10 patients, with no significant difference between groups (P = .72). Biloma occurred in 1 patient in the nonabutting group. RF ablation with electrode relocation and reduction of ablation time can be a safe and effective treatment for focal hepatic lesions adjacent to the gallbladder. Copyright © 2017 SIR. Published by Elsevier Inc. All rights reserved.

  20. Percutaneous Renal Tumor Ablation: Radiation Exposure During Cryoablation and Radiofrequency Ablation

    Energy Technology Data Exchange (ETDEWEB)

    McEachen, James C., E-mail: james.mceachen2@gmail.com [Mayo Clinic, Division of Preventive, Occupational, and Aerospace Medicine (United States); Leng, Shuai; Atwell, Thomas D. [Mayo Clinic, Department of Radiology (United States); Tollefson, Matthew K. [Mayo Clinic, Department of Urology (United States); Friese, Jeremy L. [Mayo Clinic, Department of Radiology (United States); Wang, Zhen; Murad, M. Hassan [Mayo Clinic, Division of Preventive, Occupational, and Aerospace Medicine (United States); Schmit, Grant D. [Mayo Clinic, Department of Radiology (United States)

    2016-02-15

    IntroductionOnce reserved solely for non-surgical cases, percutaneous ablation is becoming an increasingly popular treatment option for a wider array of patients with small renal masses and the radiation risk needs to be better defined as this transition continues.Materials and MethodsRetrospective review of our renal tumor ablation database revealed 425 patients who underwent percutaneous ablation for treatment of 455 renal tumors over a 5-year time period. Imparted radiation dose information was reviewed for each procedure and converted to effective patient dose and skin dose using established techniques. Statistical analysis was performed with each ablative technique.ResultsFor the 331 cryoablation procedures, the mean DLP was 6987 mGycm (SD = 2861) resulting in a mean effective dose of 104.7 mSv (SD = 43.5) and the mean CTDI{sub vol} was 558 mGy (SD = 439) resulting in a mean skin dose of 563.2 mGy (SD = 344.1). For the 124 RFA procedures, the mean DLP was 3485 mGycm (SD = 1630) resulting in a mean effective dose of 50.3 mSv (SD = 24.0) and the mean CTDI{sub vol} was 232 mGy (SD = 149) resulting in a mean skin dose of 233.2 mGy (SD = 117.4). The difference in patient radiation exposure between the two renal ablation techniques was statistically significant (p < 0.001).ConclusionBoth cryoablation and RFA imparted an average skin dose that was well below the 2 Gy deterministic threshold for appreciable sequela. Renal tumor cryoablation resulted in a mean skin and effective radiation dose more than twice that for RFA. The radiation exposure for both renal tumor ablation techniques was at the high end of the medical imaging radiation dose spectrum.

  1. PDGF-regulated rab4-dependent recycling of alphavbeta3 integrin from early endosomes is necessary for cell adhesion and spreading.

    Science.gov (United States)

    Roberts, M; Barry, S; Woods, A; van der Sluijs, P; Norman, J

    2001-09-18

    It has been postulated that the regulation of integrin vesicular traffic facilitates cell migration by internalizing integrins at the rear of the cell and transporting them forward within vesicles for exocytosis at the leading edge to form new contacts with the extracellular matrix. The rab family of GTPases control key targeting events in the endo/exocytic pathway; therefore, these GTPases may be involved in the regulation of cell-matrix contact assembly. The endo/exocytic cycle of alphavbeta3 and alpha5beta1 integrins was studied using mouse 3T3 fibroblast cell lines. In serum-starved cells, internalized integrins were transported through rab4-positive, early endosomes and arrived at the rab11-positive, perinuclear recycling compartment approximately 30 min after endocytosis. From the recycling compartment, integrins were recycled to the plasma membrane in a rab11-dependent fashion. Following treatment with PDGF, alphavbeta3 integrin, but not alpha5beta1, was rapidly recycled directly back to the plasma membrane from the early endosomes via a rab4-dependent mechanism without the involvement of rab11. This rapid recycling pathway directed alphavbeta3 to numerous small puncta distributed evenly across the dorsal surface of the cell, and the integrin only became localized into focal complexes at later times following PDGF addition. Interestingly, inhibition of PDGF-stimulated alphavbeta3 recycling using dominant-negative rab4 mutants compromised cell adhesion and spreading on vitronectin (a ligand for alphavbeta3), but adhesion to fibronectin (a ligand for alpha5beta1 and alphavbeta3) was unchanged. We propose that growth factor-regulated, rab4-dependent recycling of alphavbeta3 integrin from early endosomes to the plasma membrane is a critical upstream event in the assembly of cell-matrix contacts.

  2. Avian Influenza Virus Infection of Immortalized Human Respiratory Epithelial Cells Depends upon a Delicate Balance between Hemagglutinin Acid Stability and Endosomal pH.

    Science.gov (United States)

    Daidoji, Tomo; Watanabe, Yohei; Ibrahim, Madiha S; Yasugi, Mayo; Maruyama, Hisataka; Masuda, Taisuke; Arai, Fumihito; Ohba, Tomoyuki; Honda, Ayae; Ikuta, Kazuyoshi; Nakaya, Takaaki

    2015-04-24

    The highly pathogenic avian influenza (AI) virus, H5N1, is a serious threat to public health worldwide. Both the currently circulating H5N1 and previously circulating AI viruses recognize avian-type receptors; however, only the H5N1 is highly infectious and virulent in humans. The mechanism(s) underlying this difference in infectivity remains unclear. The aim of this study was to clarify the mechanisms responsible for the difference in infectivity between the current and previously circulating strains. Primary human small airway epithelial cells (SAECs) were transformed with the SV40 large T-antigen to establish a series of clones (SAEC-Ts). These clones were then used to test the infectivity of AI strains. Human SAEC-Ts could be broadly categorized into two different types based on their susceptibility (high or low) to the viruses. SAEC-T clones were poorly susceptible to previously circulating AI but were completely susceptible to the currently circulating H5N1. The hemagglutinin (HA) of the current H5N1 virus showed greater membrane fusion activity at higher pH levels than that of previous AI viruses, resulting in broader cell tropism. Moreover, the endosomal pH was lower in high susceptibility SAEC-T clones than that in low susceptibility SAEC-T clones. Taken together, the results of this study suggest that the infectivity of AI viruses, including H5N1, depends upon a delicate balance between the acid sensitivity of the viral HA and the pH within the endosomes of the target cell. Thus, one of the mechanisms underlying H5N1 pathogenesis in humans relies on its ability to fuse efficiently with the endosomes in human airway epithelial cells. © 2015 by The American Society for Biochemistry and Molecular Biology, Inc.

  3. Radiofrequency catheter ablation of idiopathic ventricular arrhythmias originating from intramural foci in the left ventricular outflow tract: efficacy of sequential versus simultaneous unipolar catheter ablation.

    Science.gov (United States)

    Yamada, Takumi; Maddox, William R; McElderry, H Thomas; Doppalapudi, Harish; Plumb, Vance J; Kay, G Neal

    2015-04-01

    Idiopathic ventricular arrhythmias (VAs) originating from the left ventricular outflow tract (LVOT) sometimes require catheter ablation from the endocardial and epicardial sides for their elimination, suggesting the presence of intramural VA foci. This study investigated the efficacy of sequential and simultaneous unipolar radiofrequency catheter ablation from the endocardial and epicardial sides in treating intramural LVOT VAs. Fourteen consecutive LVOT VAs, which required sequential or simultaneous irrigated unipolar radiofrequency ablation from the endocardial and epicardial sides for their elimination, were studied. The first ablation was performed at the site with the earliest local ventricular activation and best pace map on the endocardial or epicardial side. When the first ablation was unsuccessful, the second ablation was delivered on the other surface. If this sequential unipolar ablation failed, simultaneous unipolar ablation from both sides was performed. The first ablation was performed on the epicardial side in 9 VAs and endocardial side in 5 VAs. The intramural LVOT VAs were successfully eliminated by the sequential (n=9) or simultaneous (n=5) unipolar catheter ablation. Simultaneous ablation was most likely to be required for the elimination of the VAs when the distance between the endocardial and epicardial ablation sites was >8 mm and the earliest local ventricular activation time relative to the QRS onset during the VAs of sequential unipolar radiofrequency ablation and sometimes required simultaneous ablation from both the endocardial and epicardial sides. © 2015 American Heart Association, Inc.

  4. Quantitative images of metals in plant tissues measured by laser ablation inductively coupled plasma mass spectrometry

    International Nuclear Information System (INIS)

    Becker, J.S.; Dietrich, R.C.; Matusch, A.; Pozebon, D.; Dressler, V.L.

    2008-01-01

    Laser ablation inductively coupled plasma mass spectrometry (LA-ICP-MS) was used for quantitative imaging of toxic and essential elements in thin sections (thickness of 30 or 40 μm) of tobacco plant tissues. Two-dimensional images of Mg, Fe, Mn, Zn, Cu, Cd, Rh, Pt and Pb in leaves, shoots and roots of tobacco were produced. Sections of the plant tissues (fixed onto glass slides) were scanned by a focused beam of a Nd:YAG laser in a laser ablation chamber. The ablated material was transported with argon as carrier gas to the ICP ion source at a quadrupole ICP-MS instrument. Ion intensities of the investigated elements were measured together with 13 C + , 33 S + and 34 S + within the entire plant tissue section. Matrix matching standards (prepared using powder of dried tobacco leaves) were used to constitute calibration curves, whereas the regression coefficient of the attained calibration curves was typically 0.99. The variability of LA-ICP-MS process, sample heterogeneity and water content in the sample were corrected by using 13 C + as internal standard. Quantitative imaging of the selected elements revealed their inhomogeneous distribution in leaves, shoots and roots

  5. Percutaneous local ablation of unifocal subclinical breast cancer: clinical experience and preliminary results of cryotherapy

    Energy Technology Data Exchange (ETDEWEB)

    Manenti, Guglielmo; Perretta, Tommaso; Gaspari, Eleonora; Pistolese, Chiara A.; Scarano, Lia; Cossu, Elsa; Simonetti, Giovanni; Masala, Salvatore [University Hospital ' ' Tor Vergata' ' , Department of Diagnostic Imaging and Interventional Radiology, Molecular Imaging and Radiotherapy, Rome (Italy); Bonanno, Elena [University Hospital ' ' Tor Vergata' ' , Department of Biopathology, Rome (Italy); Buonomo, Oreste C.; Petrella, Giuseppe [University Hospital ' ' Tor Vergata' ' , Department of General Surgery Division, Rome (Italy)

    2011-11-15

    To assess the ablative effectiveness, the oncological and cosmetic efficacy of image-guided percutaneous cryoablation in the treatment of single breast nodules with subclinical dimensions after identification with ultrasonography (US), mammography, magnetic resonance (MRI) and characterization by vacuum assisted biopsy. Fifteen women with a mean age of 73 {+-} 5 years (range 64-82 years) and lesion diameter of 8 {+-} 4 mm were undergoing cryotherapy technology with a single probe under US-guidance associated with intra-procedural lymph-node mapping and excision of the sentinel node. All the patients underwent surgical resection (lumpectomy) from 30 to 45 days after the percutaneous ablation. The iceball size generated by the cryoprobe during the procedure at minus 40 C was 16 x 41 mm. In 14 of the 15 patients was observed a complete necrosis of the cryo-ablated lesion both in post-procedural MRI follow-up and anatomo-pathological evaluation after surgical resection. In one case there was a residual disease in post-procedural MRI and postoperative histological examination, probably justified by an incorrect positioning of the probe. The percutaneous cryoablation as a ''minimally invasive'' technique can provide excellent oncological and cosmetic results on selected cases handled by experienced operators by using the tested devices. (orig.)

  6. Percutaneous local ablation of unifocal subclinical breast cancer: clinical experience and preliminary results of cryotherapy

    International Nuclear Information System (INIS)

    Manenti, Guglielmo; Perretta, Tommaso; Gaspari, Eleonora; Pistolese, Chiara A.; Scarano, Lia; Cossu, Elsa; Simonetti, Giovanni; Masala, Salvatore; Bonanno, Elena; Buonomo, Oreste C.; Petrella, Giuseppe

    2011-01-01

    To assess the ablative effectiveness, the oncological and cosmetic efficacy of image-guided percutaneous cryoablation in the treatment of single breast nodules with subclinical dimensions after identification with ultrasonography (US), mammography, magnetic resonance (MRI) and characterization by vacuum assisted biopsy. Fifteen women with a mean age of 73 ± 5 years (range 64-82 years) and lesion diameter of 8 ± 4 mm were undergoing cryotherapy technology with a single probe under US-guidance associated with intra-procedural lymph-node mapping and excision of the sentinel node. All the patients underwent surgical resection (lumpectomy) from 30 to 45 days after the percutaneous ablation. The iceball size generated by the cryoprobe during the procedure at minus 40 C was 16 x 41 mm. In 14 of the 15 patients was observed a complete necrosis of the cryo-ablated lesion both in post-procedural MRI follow-up and anatomo-pathological evaluation after surgical resection. In one case there was a residual disease in post-procedural MRI and postoperative histological examination, probably justified by an incorrect positioning of the probe. The percutaneous cryoablation as a ''minimally invasive'' technique can provide excellent oncological and cosmetic results on selected cases handled by experienced operators by using the tested devices. (orig.)

  7. Laser ablation of a silicon target in chloroform: formation of multilayer graphite nanostructures

    Science.gov (United States)

    Abderrafi, Kamal; García-Calzada, Raúl; Sanchez-Royo, Juan F.; Chirvony, Vladimir S.; Agouram, Saïd; Abargues, Rafael; Ibáñez, Rafael; Martínez-Pastor, Juan P.

    2013-04-01

    With the use of high-resolution transmission electron microscopy, selected area electron diffraction and x-ray photoelectron spectroscopy methods of analysis we show that the laser ablation of a Si target in chloroform (CHCl3) by nanosecond UV pulses (40 ns, 355 nm) results in the formation of about 50-80 nm core-shell nanoparticles with a polycrystalline core composed of small (5-10 nm) Si and SiC mono-crystallites, the core being coated by several layers of carbon with the structure of graphite (the shell). In addition, free carbon multilayer nanostructures (carbon nano-onions) are also found in the suspension. On the basis of a comparison with similar laser ablation experiments implemented in carbon tetrachloride (CCl4), where only bare (uncoated) Si nanoparticles are produced, we suggest that a chemical (solvent decomposition giving rise to highly reactive CH-containing radicals) rather than a physical (solvent atomization followed by carbon nanostructure formation) mechanism is responsible for the formation of graphitic shells. The silicon carbonization process found for the case of laser ablation in chloroform may be promising for silicon surface protection and functionalization.

  8. PPARγ ablation sensitizes proopiomelanocortin neurons to leptin during high-fat feeding.

    Science.gov (United States)

    Long, Lihong; Toda, Chitoku; Jeong, Jing Kwon; Horvath, Tamas L; Diano, Sabrina

    2014-09-01

    Activation of central PPARγ promotes food intake and body weight gain; however, the identity of the neurons that express PPARγ and mediate the effect of this nuclear receptor on ene