Sample records for target cells pulsed

  1. Comparative study of photothermolysis of cancer cells with nuclear-targeted or cytoplasm-targeted gold nanospheres: continuous wave or pulsed lasers

    NASA Astrophysics Data System (ADS)

    Huang, Xiaohua; Kang, Bin; Qian, Wei; Mackey, Megan A.; Chen, Po C.; Oyelere, Adegboyega K.; El-Sayed, Ivan H.; El-Sayed, Mostafa A.

    2010-09-01

    We conduct a comparative study on the efficiency and cell death pathways of continuous wave (cw) and nanosecond pulsed laser photothermal cancer therapy using gold nanospheres delivered to either the cytoplasm or nucleus of cancer cells. Cytoplasm localization is achieved using arginine-glycine-aspartate peptide modified gold nanospheres, which target integrin receptors on the cell surface and are subsequently internalized by the cells. Nuclear delivery is achieved by conjugating the gold nanospheres with nuclear localization sequence peptides originating from the simian virus. Photothermal experiments show that cell death can be induced with a single pulse of a nanosecond laser more efficiently than with a cw laser. When the cw laser is applied, gold nanospheres localized in the cytoplasm are more effective in inducing cell destruction than gold nanospheres localized at the nucleus. The opposite effect is observed when the nanosecond pulsed laser is used, suggesting that plasmonic field enhancement of the nonlinear absorption processes occurs at high localization of gold nanospheres at the nucleus. Cell death pathways are further investigated via a standard apoptosis kit to show that the cell death mechanisms depend on the type of laser used. While the cw laser induces cell death via apoptosis, the nanosecond pulsed laser leads to cell necrosis. These studies add mechanistic insight to gold nanoparticle-based photothermal therapy of cancer.

  2. Target charging in short-pulse-laser-plasma experiments.

    PubMed

    Dubois, J-L; Lubrano-Lavaderci, F; Raffestin, D; Ribolzi, J; Gazave, J; Compant La Fontaine, A; d'Humières, E; Hulin, S; Nicolaï, Ph; Poyé, A; Tikhonchuk, V T

    2014-01-01

    Interaction of high-intensity laser pulses with solid targets results in generation of large quantities of energetic electrons that are the origin of various effects such as intense x-ray emission, ion acceleration, and so on. Some of these electrons are escaping the target, leaving behind a significant positive electric charge and creating a strong electromagnetic pulse long after the end of the laser pulse. We propose here a detailed model of the target electric polarization induced by a short and intense laser pulse and an escaping electron bunch. A specially designed experiment provides direct measurements of the target polarization and the discharge current in the function of the laser energy, pulse duration, and target size. Large-scale numerical simulations describe the energetic electron generation and their emission from the target. The model, experiment, and numerical simulations demonstrate that the hot-electron ejection may continue long after the laser pulse ends, enhancing significantly the polarization charge.

  3. Fc receptor-targeting of immunogen as a strategy for enhanced antigen loading, vaccination, and protection using intranasally administered antigen-pulsed dendritic cells.

    PubMed

    Pham, Giang H; Iglesias, Bibiana V; Gosselin, Edmund J

    2014-09-08

    Dendritic cells (DCs) play a critical role in the generation of adaptive immunity via the efficient capture, processing, and presentation of antigen (Ag) to naïve T cells. Administration of Ag-pulsed DCs is also an effective strategy for enhancing immunity to tumors and infectious disease organisms. Studies have also demonstrated that targeting Ags to Fcγ receptors (FcγR) on Ag presenting cells can enhance humoral and cellular immunity in vitro and in vivo. Specifically, our studies using a Francisella tularensis (Ft) infectious disease vaccine model have demonstrated that targeting immunogens to FcγR via intranasal (i.n.) administration of monoclonal antibody (mAb)-inactivated Ft (iFt) immune complexes (ICs) enhances protection against Ft challenge. Ft is the causative agent of tularemia, a debilitating disease of humans and other mammals and a category A biothreat agent for which there is no approved vaccine. Therefore, using iFt Ag as a model immunogen, we sought to determine if ex vivo targeting of iFt to FcγR on DCs would enhance the potency of i.n. administered iFt-pulsed DCs. In this study, bone marrow-derived DCs (BMDCs) were pulsed ex vivo with iFt or mAb-iFt ICs. Intranasal administration of mAb-iFt-pulsed BMDCs enhanced humoral and cellular immune responses, as well as protection against Ft live vaccine strain (LVS) challenge. Increased protection correlated with increased iFt loading on the BMDC surface as a consequence of FcγR-targeting. However, the inhibitory FcγRIIB had no impact on this enhancement. In conclusion, targeting Ag ex vivo to FcγR on DCs provides a method for enhanced Ag loading of DCs ex vivo, thereby reducing the amount of Ag required, while also avoiding the inhibitory impact of FcγRIIB. Thus, this represents a simple and less invasive strategy for increasing the potency of ex vivo-pulsed DC vaccines against chronic infectious diseases and cancer. Copyright © 2014 Elsevier Ltd. All rights reserved.

  4. Fc Receptor-Targeting of Immunogen as a Strategy for Enhanced Antigen Loading, Vaccination, and Protection Using Intranasally-Administered Antigen-Pulsed Dendritic Cells

    PubMed Central

    Pham, Giang H.; Iglesias, Bibiana V.; Gosselin, Edmund J.

    2014-01-01

    Dendritic cells (DCs) play a critical role in the generation of adaptive immunity via the efficient capture, processing, and presentation of antigen (Ag) to naïve T cells. Administration of Ag-pulsed DCs is also an effective strategy for enhancing immunity to tumors and infectious disease organisms. Studies have also demonstrated that targeting Ags to Fcγ receptors (FcγR) on Ag presenting cells can enhance humoral and cellular immunity in vitro and in vivo. Specifically, our studies using an F. tularensis (Ft) infectious disease vaccine model have demonstrated that targeting immunogens to FcγR via intranasal (i.n.) administration of monoclonal antibody (mAb)-inactivated Ft (iFt) immune complexes (ICs) enhances protection against Ft challenge. Ft is the causative agent of tularemia, a debilitating disease of humans and other mammals and a category A biothreat agent for which there is no approved vaccine. Therefore, using iFt Ag as a model immunogen, we sought to determine if ex vivo targeting of iFt to FcγR on DCs would enhance the potency of i.n. administered iFt-pulsed DCs. In this study, bone marrow-derived DCs (BMDCs) were pulsed ex vivo with iFt or mAb-iFt ICs. Intranasal administration of mAb-iFt-pulsed BMDCs enhanced humoral and cellular immune responses, as well as protection against Ft live vaccine strain (LVS) challenge. Increased protection correlated with increased iFt loading on the BMDC surface as a consequence of FcγR targeting. However, the inhibitory FcγRIIB had no impact on this enhancement. In conclusion, targeting Ag ex vivo to FcγR on DCs provides a method for enhanced Ag loading of DCs ex vivo, thereby reducing the amount of Ag required, while also avoiding the inhibitory impact of FcγRIIB. Thus, this represents a simple and less invasive strategy for increasing the potency of ex vivo-pulsed DC vaccines against chronic infectious diseases and cancer. PMID:25068496

  5. Influence of lateral target size on hot electron production and electromagnetic pulse emission from laser-irradiated metallic targets

    NASA Astrophysics Data System (ADS)

    Chen, Zi-Yu; Li, Jian-Feng; Yu, Yong; Wang, Jia-Xiang; Li, Xiao-Ya; Peng, Qi-Xian; Zhu, Wen-Jun

    2012-11-01

    The influences of lateral target size on hot electron production and electromagnetic pulse emission from laser interaction with metallic targets have been investigated. Particle-in-cell simulations at high laser intensities show that the yield of hot electrons tends to increase with lateral target size, because the larger surface area reduces the electrostatic field on the target, owing to its expansion along the target surface. At lower laser intensities and longer time scales, experimental data characterizing electromagnetic pulse emission as a function of lateral target size also show target-size effects. Charge separation and a larger target tending to have a lower target potential have both been observed. The increase in radiation strength and downshift in radiation frequency with increasing lateral target size can be interpreted using a simple model of the electrical capacity of the target.

  6. Targeted cellular ablation based on the morphology of malignant cells

    NASA Astrophysics Data System (ADS)

    Ivey, Jill W.; Latouche, Eduardo L.; Sano, Michael B.; Rossmeisl, John H.; Davalos, Rafael V.; Verbridge, Scott S.

    2015-11-01

    Treatment of glioblastoma multiforme (GBM) is especially challenging due to a shortage of methods to preferentially target diffuse infiltrative cells, and therapy-resistant glioma stem cell populations. Here we report a physical treatment method based on electrical disruption of cells, whose action depends strongly on cellular morphology. Interestingly, numerical modeling suggests that while outer lipid bilayer disruption induced by long pulses (~100 μs) is enhanced for larger cells, short pulses (~1 μs) preferentially result in high fields within the cell interior, which scale in magnitude with nucleus size. Because enlarged nuclei represent a reliable indicator of malignancy, this suggested a means of preferentially targeting malignant cells. While we demonstrate killing of both normal and malignant cells using pulsed electric fields (PEFs) to treat spontaneous canine GBM, we proposed that properly tuned PEFs might provide targeted ablation based on nuclear size. Using 3D hydrogel models of normal and malignant brain tissues, which permit high-resolution interrogation during treatment testing, we confirmed that PEFs could be tuned to preferentially kill cancerous cells. Finally, we estimated the nuclear envelope electric potential disruption needed for cell death from PEFs. Our results may be useful in safely targeting the therapy-resistant cell niches that cause recurrence of GBM tumors.

  7. Targeted cellular ablation based on the morphology of malignant cells

    PubMed Central

    Ivey, Jill W.; Latouche, Eduardo L.; Sano, Michael B.; Rossmeisl, John H.; Davalos, Rafael V.; Verbridge, Scott S.

    2015-01-01

    Treatment of glioblastoma multiforme (GBM) is especially challenging due to a shortage of methods to preferentially target diffuse infiltrative cells, and therapy-resistant glioma stem cell populations. Here we report a physical treatment method based on electrical disruption of cells, whose action depends strongly on cellular morphology. Interestingly, numerical modeling suggests that while outer lipid bilayer disruption induced by long pulses (~100 μs) is enhanced for larger cells, short pulses (~1 μs) preferentially result in high fields within the cell interior, which scale in magnitude with nucleus size. Because enlarged nuclei represent a reliable indicator of malignancy, this suggested a means of preferentially targeting malignant cells. While we demonstrate killing of both normal and malignant cells using pulsed electric fields (PEFs) to treat spontaneous canine GBM, we proposed that properly tuned PEFs might provide targeted ablation based on nuclear size. Using 3D hydrogel models of normal and malignant brain tissues, which permit high-resolution interrogation during treatment testing, we confirmed that PEFs could be tuned to preferentially kill cancerous cells. Finally, we estimated the nuclear envelope electric potential disruption needed for cell death from PEFs. Our results may be useful in safely targeting the therapy-resistant cell niches that cause recurrence of GBM tumors. PMID:26596248

  8. Strong electromagnetic pulses generated in high-intensity short-pulse laser interactions with thin foil targets

    NASA Astrophysics Data System (ADS)

    Rączka, P.; Dubois, J.-L.; Hulin, S.; Tikhonchuk, V.; Rosiński, M.; Zaraś-Szydłowska, A.; Badziak, J.

    2017-12-01

    Measurements are reported of the target neutralization current, the target charge, and the tangential component of the magnetic field generated as a result of laser-target interaction by pulses with the energy in the range of 45 mJ to 92 mJ on target and the pulse duration from 39 fs to 1000 fs. The experiment was performed at the Eclipse facility in CELIA, Bordeaux. The aim of the experiment was to extend investigations performed for the thick (mm scale) targets to the case of thin (micrometer thickness) targets in a way that would allow for a straightforward comparison of the results. We found that thin foil targets tend to generate 20 to 50 percent higher neutralization current and the target charge than the thick targets. The measurement of the tangential component of the magnetic field had shown that the initial spike is dominated by the 1 ns pulse consistent with the 1 ns pulse of the neutralization current, but there are some differences between targets of different type on sub-ns scale, which is an effect going beyond a simple picture of the target acting as an antenna. The sub-ns structure appears to be reproducible to surprising degree. We found that there is in general a linear correlation between the maximum value of the magnetic field and the maximum neutralization current, which supports the target-antenna picture, except for pulses hundreds of fs long.

  9. Nurse opinions and pulse oximeter saturation target limits for preterm infants.

    PubMed

    Nghiem, Tuyet-Hang; Hagadorn, James I; Terrin, Norma; Syke, Sally; MacKinnon, Brenda; Cole, Cynthia H

    2008-05-01

    The objectives of this study were to compare pulse oximeter saturation limits targeted by nurses for extremely preterm infants during routine care with nurse opinions regarding appropriate pulse oximeter saturation limits and with policy-specified pulse oximeter saturation limits and to identify factors that influence pulse oximeter saturation limits targeted by nurses. We surveyed nurses in US NICUs with neonatal-perinatal fellowships in 2004. Data collected included pulse oximeter saturation limits targeted by nurses and by NICU policy when present, nurses' opinions about appropriate pulse oximeter saturation limits, and NICU and nurse characteristics. Factors associated with pulse oximeter saturation limits targeted by nurses were identified with hierarchical linear modeling. Among those eligible, 2805 (45%) nurses in 59 (60%) NICUs responded. Forty (68%) of 59 NICUs had a policy that specified a pulse oximeter saturation target range for extremely preterm infants. Among 1957 nurses at NICUs with policies, 540 (28%) accurately identified the upper and lower limits of their NICU's policy and also targeted these values in practice. NICU-specific SDs for individual nurse target limits were less at NICUs with versus without a policy for both upper and lower limits. Hierarchical linear modeling identified presence of pulse oximeter saturation policy, NICU-specific nurse group opinion, and individual nurse opinion as factors significantly associated with individual pulse oximeter saturation target limits. For each percentage point increase in individual opinion upper limit, the individual target upper limit increased by 0.41 percentage point at NICUs with a policy compared with 0.6 percentage point at NICUs with no policy. Presence of policy-specified pulse oximeter saturation limits, nurse group opinion, and individual nurse opinion were independently associated with individual nurse pulse oximeter saturation target limits during routine care of extremely preterm

  10. Atmospheric electromagnetic pulse propagation effects from thick targets in a terawatt laser target chamber.

    PubMed

    Remo, John L; Adams, Richard G; Jones, Michael C

    2007-08-20

    Generation and effects of atmospherically propagated electromagnetic pulses (EMPs) initiated by photoelectrons ejected by the high density and temperature target surface plasmas from multiterawatt laser pulses are analyzed. These laser radiation pulse interactions can significantly increase noise levels, thereby obscuring data (sometimes totally) and may even damage sensitive probe and detection instrumentation. Noise effects from high energy density (approximately multiterawatt) laser pulses (approximately 300-400 ps pulse widths) interacting with thick approximately 1 mm) metallic and dielectric solid targets and dielectric-metallic powder mixtures are interpreted as transient resonance radiation associated with surface charge fluctuations on the target chamber that functions as a radiating antenna. Effective solutions that minimize atmospheric EMP effects on internal and proximate electronic and electro-optical equipment external to the system based on systematic measurements using Moebius loop antennas, interpretations of signal periodicities, and dissipation indicators determining transient noise origin characteristics from target emissions are described. Analytic models for the effect of target chamber resonances and associated noise current and temperature in a probe diode laser are described.

  11. Atmospheric electromagnetic pulse propagation effects from thick targets in a terawatt laser target chamber

    NASA Astrophysics Data System (ADS)

    Remo, John L.; Adams, Richard G.; Jones, Michael C.

    2007-08-01

    Generation and effects of atmospherically propagated electromagnetic pulses (EMPs) initiated by photoelectrons ejected by the high density and temperature target surface plasmas from multiterawatt laser pulses are analyzed. These laser radiation pulse interactions can significantly increase noise levels, thereby obscuring data (sometimes totally) and may even damage sensitive probe and detection instrumentation. Noise effects from high energy density (approximately multiterawatt) laser pulses (˜300-400 ps pulse widths) interacting with thick (˜1 mm) metallic and dielectric solid targets and dielectric-metallic powder mixtures are interpreted as transient resonance radiation associated with surface charge fluctuations on the target chamber that functions as a radiating antenna. Effective solutions that minimize atmospheric EMP effects on internal and proximate electronic and electro-optical equipment external to the system based on systematic measurements using Moebius loop antennas, interpretations of signal periodicities, and dissipation indicators determining transient noise origin characteristics from target emissions are described. Analytic models for the effect of target chamber resonances and associated noise current and temperature in a probe diode laser are described.

  12. Atmospheric electromagnetic pulse propagation effects from thick targets in a terawatt laser target chamber

    DOE PAGES

    Remo, John L.; Adams, Richard G.; Jones, Michael C.

    2007-08-16

    Generation and effects of atmospherically propagated electromagnetic pulses (EMPs) initiated by photoelectrons ejected by the high density and temperature target surface plasmas from multiterawatt laser pulses are analyzed. These laser radiation pulse interactions can significantly increase noise levels, thereby obscuring data (sometimes totally) and may even damage sensitive probe and detection instrumentation. Noise effects from high energy density (approximately multiterawatt) laser pulses (~300–400 ps pulse widths) interacting with thick (~1 mm) metallic and dielectric solid targets and dielectric–metallic powder mixtures are interpreted as transient resonance radiation associated with surface charge fluctuations on the target chamber that functions as a radiatingmore » antenna. Effective solutions that minimize atmospheric EMP effects on internal and proximate electronic and electro-optical equipment external to the system based on systematic measurements using Moebius loop antennas, interpretations of signal periodicities, and dissipation indicators determining transient noise origin characteristics from target emissions are described. Analytic models for the effect of target chamber resonances and associated noise current and temperature in a probe diode laser are described.« less

  13. Heat pulse excitability of vestibular hair cells and afferent neurons

    PubMed Central

    Brichta, Alan M.; Tabatabaee, Hessam; Boutros, Peter J.; Ahn, JoongHo; Della Santina, Charles C.; Poppi, Lauren A.; Lim, Rebecca

    2016-01-01

    In the present study we combined electrophysiology with optical heat pulse stimuli to examine thermodynamics of membrane electrical excitability in mammalian vestibular hair cells and afferent neurons. We recorded whole cell currents in mammalian type II vestibular hair cells using an excised preparation (mouse) and action potentials (APs) in afferent neurons in vivo (chinchilla) in response to optical heat pulses applied to the crista (ΔT ≈ 0.25°C per pulse). Afferent spike trains evoked by heat pulse stimuli were diverse and included asynchronous inhibition, asynchronous excitation, and/or phase-locked APs synchronized to each infrared heat pulse. Thermal responses of membrane currents responsible for APs in ganglion neurons were strictly excitatory, with Q10 ≈ 2. In contrast, hair cells responded with a mix of excitatory and inhibitory currents. Excitatory hair cell membrane currents included a thermoelectric capacitive current proportional to the rate of temperature rise (dT/dt) and an inward conduction current driven by ΔT. An iberiotoxin-sensitive inhibitory conduction current was also evoked by ΔT, rising in <3 ms and decaying with a time constant of ∼24 ms. The inhibitory component dominated whole cell currents in 50% of hair cells at −68 mV and in 67% of hair cells at −60 mV. Responses were quantified and described on the basis of first principles of thermodynamics. Results identify key molecular targets underlying heat pulse excitability in vestibular sensory organs and provide quantitative methods for rational application of optical heat pulses to examine protein biophysics and manipulate cellular excitability. PMID:27226448

  14. Enhanced proton acceleration from an ultrathin target irradiated by laser pulses with plateau ASE.

    PubMed

    Wang, Dahui; Shou, Yinren; Wang, Pengjie; Liu, Jianbo; Li, Chengcai; Gong, Zheng; Hu, Ronghao; Ma, Wenjun; Yan, Xueqing

    2018-02-07

    We report a simulation study on proton acceleration driven by ultraintense laser pulses with normal contrast (10 7 -10 9 ) containing nanosecond plateau amplified spontaneous emission (ASE). It's found in hydrodynamic simulations that if the thickness of the targets lies in the range of hundreds nanometer matching the intensity and duration of ASE, the ablation pressure would push the whole target in the forward direction with speed exceeding the expansion velocity of plasma, resulting in a plasma density profile with a long extension at the target front and a sharp gradient at the target rear. When the main pulse irradiates the plasma, self-focusing happens at the target front, producing highly energetic electrons through direct laser acceleration(DLA) building the sheath field. The sharp plasma gradient at target rear ensures a strong sheath field. 2D particle-in-cell(PIC) simulations reveal that the proton energy can be enhanced by a factor of 2 compared to the case of using micrometer-thick targets.

  15. Dielectrophoretic focusing integrated pulsed laser activated cell sorting

    NASA Astrophysics Data System (ADS)

    Zhu, Xiongfeng; Kung, Yu-Chun; Wu, Ting-Hsiang; Teitell, Michael A.; Chiou, Pei-Yu

    2017-08-01

    We present a pulsed laser activated cell sorter (PLACS) integrated with novel sheathless size-independent dielectrophoretic (DEP) focusing. Microfluidic fluorescence activated cell sorting (μFACS) systems aim to provide a fully enclosed environment for sterile cell sorting and integration with upstream and downstream microfluidic modules. Among them, PLACS has shown a great potential in achieving comparable performance to commercial aerosol-based FACS (>90% purity at 25,000 cells sec-1). However conventional sheath flow focusing method suffers a severe sample dilution issue. Here we demonstrate a novel dielectrophoresis-integrated pulsed laser activated cell sorter (DEP-PLACS). It consists of a microfluidic channel with 3D electrodes laid out to provide a tunnel-shaped electric field profile along a 4cmlong channel for sheathlessly focusing microparticles/cells into a single stream in high-speed microfluidic flows. All focused particles pass through the fluorescence detection zone along the same streamline regardless of their sizes and types. Upon detection of target fluorescent particles, a nanosecond laser pulse is triggered and focused in a neighboring channel to generate a rapidly expanding cavitation bubble for precise sorting. DEP-PLACS has achieved a sorting purity of 91% for polystyrene beads at a throughput of 1,500 particle/sec.

  16. Self-calibrating d-scan: measuring ultrashort laser pulses on-target using an arbitrary pulse compressor.

    PubMed

    Alonso, Benjamín; Sola, Íñigo J; Crespo, Helder

    2018-02-19

    In most applications of ultrashort pulse lasers, temporal compressors are used to achieve a desired pulse duration in a target or sample, and precise temporal characterization is important. The dispersion-scan (d-scan) pulse characterization technique usually involves using glass wedges to impart variable, well-defined amounts of dispersion to the pulses, while measuring the spectrum of a nonlinear signal produced by those pulses. This works very well for broadband few-cycle pulses, but longer, narrower bandwidth pulses are much more difficult to measure this way. Here we demonstrate the concept of self-calibrating d-scan, which extends the applicability of the d-scan technique to pulses of arbitrary duration, enabling their complete measurement without prior knowledge of the introduced dispersion. In particular, we show that the pulse compressors already employed in chirped pulse amplification (CPA) systems can be used to simultaneously compress and measure the temporal profile of the output pulses on-target in a simple way, without the need of additional diagnostics or calibrations, while at the same time calibrating the often-unknown differential dispersion of the compressor itself. We demonstrate the technique through simulations and experiments under known conditions. Finally, we apply it to the measurement and compression of 27.5 fs pulses from a CPA laser.

  17. Hypericin and pulsed laser therapy of squamous cell cancer in vitro.

    PubMed

    Bublik, Michael; Head, Christian; Benharash, Peyman; Paiva, Marcos; Eshraghi, Adrian; Kim, Taiho; Saxton, Romaine

    2006-06-01

    This in vitro study compares continuous wave and pulsed laser light at longer wavelengths for activation of the phototoxic drug hypericin in human cancer cells. Two-photon pulsed laser light now allows high-resolution fluorescent imaging of cancer cells and should provide deeper tissue penetration with near infrared light for improved detection as well as phototoxicity in human tumors. Cultured Seoul National University (SNU)-1 tumor cells from a squamous cell carcinoma (SCC) were incubated with hypericin before photoirradiation at four laser wavelengths. Phototoxicity of hypericin sensitized SCC cells was measured by dimethyl thiazoldiphenyl (MTT) tetrazolium bromide cell viability assays and by confocal fluorescence microscopy via 532-nm and infrared two-photon pulsed laser light. Phototoxic response increased linearly with hypericin dose of 0.1-2 microM, light exposure time of 5-120 sec, and pulsed dye laser wavelengths of 514-593 nm. Light energy delivery for 50% cell phototoxicity (LD50) response was 9 joules at 514 nm, 3 joules at 550 nm, and less than 1 joule at the 593 nm hypericin light absorption maxima. Fluorescence confocal microscopy revealed membrane and perinuclear localization of hypericin in the SNU cells with membrane damage seen after excitation with visible 532 nm continuous wave light or two-photon 700-950 nm picosecond pulsed laser irradiation. Hypericin may be a powerful tumor targetting drug when combined with pulsed laser light in patients with recurrent head and neck SCC.

  18. Monoenergetic ion acceleration and Rayleigh-Taylor instability of the composite target irradiated by the laser pulse

    NASA Astrophysics Data System (ADS)

    Khudik, Vladimir; Yi, S. Austin; Shvets, Gennady

    2012-10-01

    Acceleration of ions in the two-specie composite target irradiated by a circularly polarized laser pulse is studied analytically and via particle-in-cell (PIC) simulations. A self-consistent analytical model of the composite target is developed. In this model, target parameters are stationary in the center of mass of the system: heavy and light ions are completely separated from each other and form two layers, while electrons are bouncing in the potential well formed by the laser ponderomotive and electrostatic potentials. They are distributed in the direction of acceleration by the Boltzmann law and over velocities by the Maxwell-Juttner law. The laser pulse interacts directly only with electrons in a thin sheath layer, and these electrons transfer the laser pressure to the target ions. In the fluid approximation it is shown, the composite target is still susceptible to the Rayleigh-Taylor instability [1]. Using PIC simulations we found the growth rate of initially seeded perturbations as a function of their wavenumber for different composite target parameters and compare it with analytical results. Useful scaling laws between this rate and laser pulse pressure and target parameters are discussed.[4pt] [1] T.P. Yu, A. Pukhov, G. Shvets, M. Chen, T. H. Ratliff, S. A. Yi, and V. Khudik, Phys. Plasmas, 18, 043110 (2011).

  19. Heating of solid targets with laser pulses

    NASA Technical Reports Server (NTRS)

    Bechtel, J. H.

    1975-01-01

    Analytical and numerical solutions to the heat-conduction equation are obtained for the heating of absorbing media with pulsed lasers. The spatial and temporal form of the temperature is determined using several different models of the laser irradiance. Both surface and volume generation of heat are discussed. It is found that if the depth of thermal diffusion for the laser-pulse duration is large compared to the optical-attenuation depth, the surface- and volume-generation models give nearly identical results. However, if the thermal-diffusion depth for the laser-pulse duration is comparable to or less than the optical-attenuation depth, the surface-generation model can give significantly different results compared to the volume-generation model. Specific numerical results are given for a tungsten target irradiated by pulses of different temporal durations and the implications of the results are discussed with respect to the heating of metals by picosecond laser pulses.

  20. Targeted disruption of deep-lying neocortical microvessels in rat using ultrashort laser pulses

    NASA Astrophysics Data System (ADS)

    Nishimura, Nozomi; Schaffer, Christopher B.; Friedman, Beth; Tsai, Philbert S.; Lyden, Patrick D.; Kleinfeld, David

    2004-06-01

    The study of neurovascular diseases such as vascular dementia and stroke require novel models of targeted vascular disruption in the brain. We describe a model of microvascular disruption in rat neocortex that uses ultrashort laser pulses to induce localized injury to specific targeted microvessels and uses two-photon microscopy to monitor and guide the photodisruption process. In our method, a train of high-intensity, 100-fs laser pulses is tightly focused into the lumen of a blood vessel within the upper 500 μm of cortex. Photodisruption induced by these laser pulses creates injury to a single vessel located at the focus of the laser, leaving the surrounding tissue intact. This photodisruption results in three modalities of localized vascular injury. At low power, blood plasma extravasation can be induced. The vessel itself remains intact, while serum is extravasated into the intercellular space. Localized ischemia caused by an intravascular clot results when the photodisruption leads to a brief disturbance of the vascular walls that initiates an endogenous clotting cascade. The formation of a localized thrombus stops the blood flow at the location of the photodisruption. A hemorrhage, defined as a large extravasation of blood including plasma and red blood cells, results when higher laser power is used. The targeted vessel does not remain intact.

  1. Pulsed Laser Illumination of Photovoltaic Cells

    NASA Technical Reports Server (NTRS)

    Yater, Jane A.; Lowe, Roland; Jenkins, Philip; Landis, Geoffrey A.

    1994-01-01

    In future space missions, free electron lasers (FEL) may be used to illuminate photovoltaic array receivers to provide remote power. The induction FEL and the radio-frequency (RF) FEL both produce pulsed rather than continuous output. In this work, we investigate cell response to pulsed laser light which simulates the RF FEL format, producing 50 ps pulses at a frequency of 78 MHz. A variety of Si, GaAs, CaSb and CdInSe2 (CIS) solar cells are tested at average incident powers between 4 mW/sq cm and 425 mW/sq cm. The results indicate that if the pulse repetition is high, cell efficiencies are only slightly reduced by using a pulsed laser source compared to constant illumination at the same wavelength. Because the pulse separation is less than or approximately equal to the minority carrier lifetime, the illumination conditions are effectively those of a continuous wave laser. The time dependence of the voltage and current response of the cells are also measured using a sampling oscilloscope equipped with a high frequency voltage probe and current transformer. The frequency response of the cells is weak, with both voltage and current outputs essentially dc in nature. Comparison with previous experiments shows that the RF FEL pulse format yields much more efficient photovoltaic conversion of light than does an induction FEL pulse format.

  2. Pulsed laser illumination of photovoltaic cells

    NASA Technical Reports Server (NTRS)

    Yater, Jane A.; Lowe, Roland A.; Jenkins, Phillip P.; Landis, Geoffrey A.

    1994-01-01

    In future space missions, free electron lasers (FEL) may be used to illuminate photovoltaic array receivers to provide remote power. Both the radio-frequency (RF) and induction FEL provide FEL produce pulsed rather than continuous output. In this work we investigate cell response to pulsed laser light which simulates the RF FEL format. The results indicate that if the pulse repetition is high, cell efficiencies are only slightly reduced compared to constant illumination at the same wavelength. The frequency response of the cells is weak, with both voltage and current outputs essentially dc in nature. Comparison with previous experiments indicates that the RF FEL pulse format yields more efficient photovoltaic conversion than does an induction FEL pulse format.

  3. Pulse charging of lead-acid traction cells

    NASA Technical Reports Server (NTRS)

    Smithrick, J. J.

    1980-01-01

    Pulse charging, as a method of rapidly and efficiently charging 300 amp-hour lead-acid traction cells for an electric vehicle application was investigated. A wide range of charge pulse current square waveforms were investigated and the results were compared to constant current charging at the time averaged pulse current values. Representative pulse current waveforms were: (1) positive waveform-peak charge pulse current of 300 amperes (amps), discharge pulse-current of zero amps, and a duty cycle of about 50%; (2) Romanov waveform-peak charge pulse current of 300 amps, peak discharge pulse current of 15 amps, and a duty of 50%; and (3) McCulloch waveform peak charge pulse current of 193 amps, peak discharge pulse current of about 575 amps, and a duty cycle of 94%. Experimental results indicate that on the basis of amp-hour efficiency, pulse charging offered no significant advantage as a method of rapidly charging 300 amp-hour lead-acid traction cells when compared to constant current charging at the time average pulse current value. There were, however, some disadvantages of pulse charging in particular a decrease in charge amp-hour and energy efficiencies and an increase in cell electrolyte temperature. The constant current charge method resulted in the best energy efficiency with no significant sacrifice of charge time or amp-hour output. Whether or not pulse charging offers an advantage over constant current charging with regard to the cell charge/discharge cycle life is unknown at this time.

  4. Pulsed laser illumination of photovoltaic cells

    NASA Technical Reports Server (NTRS)

    Yater, Jane A.; Lowe, Roland A.; Jenkins, Phillip P.; Landis, Geoffrey A.

    1995-01-01

    In future space missions, free electron lasers (FEL) may be used to illuminate photovoltaic receivers to provide remote power. Both the radio-frequency (RF) and induction FEL produce pulsed rather than continuous output. In this work we investigate cell response to pulsed laser light which simulates the RF FEL format. The results indicate that if the pulse repetition is high, cell efficiencies are only slightly reduced compared to constant illumination at the same wavelength. The frequency response of the cells is weak, with both voltage and current outputs essentially dc in nature. Comparison with previous experiments indicates that the RF FEL pulse format yields more efficient photovoltaic conversion than does an induction FEL format.

  5. Cell-Specific Multifunctional Processing of Heterogeneous Cell Systems in a Single Laser Pulse Treatment

    PubMed Central

    Lukianova-Hleb, Ekaterina Y.; Mutonga, Martin B. G.; Lapotko, Dmitri O.

    2012-01-01

    Current methods of cell processing for gene and cell therapies use several separate procedures for gene transfer and cell separation or elimination, because no current technology can offer simultaneous multi-functional processing of specific cell sub-sets in highly heterogeneous cell systems. Using the cell-specific generation of plasmonic nanobubbles of different sizes around cell-targeted gold nanoshells and nanospheres, we achieved simultaneous multifunctional cell-specific processing in a rapid single 70 ps laser pulse bulk treatment of heterogeneous cell suspension. This method supported the detection of cells, delivery of external molecular cargo to one type of cells and the concomitant destruction of another type of cells without damaging other cells in suspension, and real-time guidance of the two above cellular effects. PMID:23167546

  6. Manifestation of anharmonic resonance in the interaction of intense ultrashort laser pulses with microstructured targets

    NASA Astrophysics Data System (ADS)

    Dalui, Malay; Kundu, M.; Madhu Trivikram, T.; Ray, Krishanu; Krishnamurthy, M.

    2016-10-01

    Identification of the basic processes responsible for an efficient heating of intense laser produced plasmas is one of the important features of high intensity laser matter interaction studies. Collisionless absorption due to the anharmonicity in the self-consistent electrostatic potential of the plasma, known as anharmonic resonance (AHR), has been proposed to be a basic mechanism but a clear experimental demonstration is needed. Here, we show that microstructured targets enhance X-ray emission and the polarization dependence ascribes the enhancement to anharmonic resonance heating. It is found that p-polarized pulses of 5 ×1017 W/cm2 intensity bring in a 16-fold enhancement in the X-ray emission in the energy range 20-350 keV compared to s-polarized pulses with microstructured targets. This ratio is 2 for the case of polished targets under otherwise identical conditions. Particle-in-cell simulations clearly show that AHR is the key absorption mechanism responsible for this effect.

  7. Killing of targets by effector CD8 T cells in the mouse spleen follows the law of mass action

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    Ganusov, Vitaly V

    2009-01-01

    In contrast with antibody-based vaccines, it has been difficult to measure the efficacy of T cell-based vaccines and to correlate the efficacy of CD8 T cell responses with protection again viral infections. In part, this difficulty is due to poor understanding of the in vivo efficacy of CD8 T cells produced by vaccination. Using a: recently developed experimental method of in vivo cytotoxicity we have investigated quantitative aspects of killing of peptide-pulsed targets by effector and memory CD8 T cells, specific to three epitopes of lymphocytic choriomeningitis virus (LCMV), in the mouse spleen. By analyzing data on killing of targetsmore » with varying number of epitope-specific effector and memory CD8 T cells, we find that killing of targets by effectors follows the law of mass-action, that is the death rate of peptide-pulsed targets is proportional to the frequency of CTLs in the spleen. In contrast, killing of targets by memory CD8 T cells does not follow the mass action law because the death rate of targets saturates at high frequencies of memory CD8 T cells. For both effector and memory cells, we also find little support for the killing term that includes the decrease of the death rate of targets with target cell density. Interestingly, our analysis suggests that at low CD8 T cell frequencies, memory CD8 T cells on the per capita basis are more efficient at killing peptide-pulsed targets than effectors, but at high frequencies, effectors are more efficient killers than memory T cells. Comparison of the estimated killing efficacy of effector T cells with the value that is predicted from theoretical physics and based on motility of T cells in lymphoid tissues, suggests that limiting step in the killing of peptide-pulsed targets is delivering the lethal hit and not finding the target. Our results thus form a basis for quantitative understanding of the process of killing of virus-infected cells by T cell responses in tissues and can be used to correlate the

  8. Antigen targeting to antigen-presenting cells enhances presentation to class II-restricted T lymphocytes.

    PubMed Central

    Scardino, A; Paroli, M; De Petrillo, G; Michel, M L; Barnaba, V

    1994-01-01

    Receptor-mediated uptake increases by several orders of magnitude the efficiency of APC to internalize Ag, and is stringently required for the Ag-presenting function of T lymphocytes due to their inability to take up Ag non-specifically. We have previously reported that hepatitis B envelope antigen (HBenvAg) can be internalized by T cells via transferrin receptor (TfR). To evaluate if Ag targeting to receptors expressed on APC could be an effective tool for promoting Ag uptake and presentation, we tested the capacity of activated T cells not expressing TfR to induce HBenvAg-specific T-cell responses when pulsed with a hybrid particle containing HBenvAg coupled to gp120 of human immunodeficiency virus (HIV), exploiting the ability of gp120 to bind to CD4 receptor. We found that CD4+/TfR- T cells pulsed either with the hybrid particle or peptide (S193-207) but not with S, L Ag, a recombinant form of HBenvAg, induced a specific proliferative response of a T-cell clone recognizing peptide (S193-207) of HBenvAg. The finding that the addition of anti-CD4 monoclonal antibody (mAb) before the pulsing of CD4+/TfR- T cells with the hybrid particle drastically blocked the specific T-cell response, together with the finding that CD8+/TfR- T cells were unable to serve as APC even if pulsed with this molecule, demonstrated that CD4 receptor was crucial for the HBenvAg internalization. On the other hand, HBenvAg presentation by CD4+/TfR+ T cells pulsed with the hybrid particle was inhibited only when both anti-CD4 and anti-TfR were added before the pulsing. These results suggest that Ag targeting to APC receptors may be usefully exploited to improve Ag-presentation efficiency in potential immunotherapeutic approaches. PMID:7907575

  9. Shock ion acceleration by an ultrashort circularly polarized laser pulse via relativistic transparency in an exploded target.

    PubMed

    Kim, Young-Kuk; Cho, Myung-Hoon; Song, Hyung Seon; Kang, Teyoun; Park, Hyung Ju; Jung, Moon Youn; Hur, Min Sup

    2015-10-01

    We investigated ion acceleration by an electrostatic shock in an exploded target irradiated by an ultrashort, circularly polarized laser pulse by means of one- and three-dimensional particle-in-cell simulations. We discovered that the laser field penetrating via relativistic transparency (RT) rapidly heated the upstream electron plasma to enable the formation of a high-speed electrostatic shock. Owing to the RT-based rapid heating and the fast compression of the initial density spike by a circularly polarized pulse, a new regime of the shock ion acceleration driven by an ultrashort (20-40 fs), moderately intense (1-1.4 PW) laser pulse is envisaged. This regime enables more efficient shock ion acceleration under a limited total pulse energy than a linearly polarized pulse with crystal laser systems of λ∼1μm.

  10. Near-IR laser-triggered target cell collection using a carbon nanotube-based cell-cultured substrate.

    PubMed

    Sada, Takao; Fujigaya, Tsuyohiko; Niidome, Yasuro; Nakazawa, Kohji; Nakashima, Naotoshi

    2011-06-28

    Unique near-IR optical properties of single-walled carbon nanotube (SWNTs) are of interest in many biological applications. Here we describe the selective cell detachment and collection from an SWNT-coated cell-culture dish triggered by near-IR pulse laser irradiation. First, HeLa cells were cultured on an SWNT-coated dish prepared by a spraying of an aqueous SWNT dispersion on a glass dish. The SWNT-coated dish was found to show a good cell adhesion behavior as well as a cellular proliferation rate similar to a conventional glass dish. We discovered, by near-IR pulse laser irradiation (at the laser power over 25 mW) to the cell under optical microscopic observation, a quick single-cell detachment from the SWNT-coated surface. Shockwave generation from the irradiated SWNTs is expected to play an important role for the cell detachment. Moreover, we have succeeded in catapulting the target single cell from the cultured medium when the depth of the medium was below 150 μm and the laser power was stronger than 40 mW. The captured cell maintained its original shape. The retention of the genetic information of the cell was confirmed by the polymerase chain reaction (PCR) technique. A target single-cell collection from a culture medium under optical microscopic observation is significant in wide fields of single-cell studies in biological areas.

  11. Effects of high voltage nanosecond electric pulses on eukaryotic cells (in vitro): A systematic review.

    PubMed

    Batista Napotnik, Tina; Reberšek, Matej; Vernier, P Thomas; Mali, Barbara; Miklavčič, Damijan

    2016-08-01

    For this systematic review, 203 published reports on effects of electroporation using nanosecond high-voltage electric pulses (nsEP) on eukaryotic cells (human, animal, plant) in vitro were analyzed. A field synopsis summarizes current published data in the field with respect to publication year, cell types, exposure configuration, and pulse duration. Published data were analyzed for effects observed in eight main target areas (plasma membrane, intracellular, apoptosis, calcium level and distribution, survival, nucleus, mitochondria, stress) and an additional 107 detailed outcomes. We statistically analyzed effects of nsEP with respect to three pulse duration groups: A: 1-10ns, B: 11-100ns and C: 101-999ns. The analysis confirmed that the plasma membrane is more affected with longer pulses than with short pulses, seen best in uptake of dye molecules after applying single pulses. Additionally, we have reviewed measurements of nsEP and evaluations of the electric fields to which cells were exposed in these reports, and we provide recommendations for assessing nanosecond pulsed electric field effects in electroporation studies. Copyright © 2016 The Authors. Published by Elsevier B.V. All rights reserved.

  12. Nanosurgery of cells and chromosomes using near-infrared twelve-femtosecond laser pulses

    NASA Astrophysics Data System (ADS)

    Uchugonova, Aisada; Lessel, Matthias; Nietzsche, Sander; Zeitz, Christian; Jacobs, Karin; Lemke, Cornelius; König, Karsten

    2012-10-01

    Laser-assisted surgery based on multiphoton absorption of near-infrared laser light has great potential for high precision surgery at various depths within the cells and tissues. Clinical applications include refractive surgery (fs-LASIK). The non-contact laser method also supports contamination-free cell nanosurgery. In this paper we describe usage of an ultrashort femtosecond laser scanning microscope for sub-100 nm surgery of human cells and metaphase chromosomes. A mode-locked 85 MHz Ti:Sapphire laser with an M-shaped ultrabroad band spectrum (maxima: 770 nm/830 nm) and an in situ pulse duration at the target ranging from 12 fs up to 3 ps was employed. The effects of laser nanoprocessing in cells and chromosomes have been quantified by atomic force microscopy. These studies demonstrate the potential of extreme ultrashort femtosecond laser pulses at low mean milliwatt powers for sub-100 nm surgery of cells and cellular organelles.

  13. Modeling of intense pulsed ion beam heated masked targets for extreme materials characterization

    NASA Astrophysics Data System (ADS)

    Barnard, John J.; Schenkel, Thomas

    2017-11-01

    Intense, pulsed ion beams locally heat materials and deliver dense electronic excitations that can induce material modifications and phase transitions. Material properties can potentially be stabilized by rapid quenching. Pulsed ion beams with pulse lengths of order ns have recently become available for materials processing. Here, we optimize mask geometries for local modification of materials by intense ion pulses. The goal is to rapidly excite targets volumetrically to the point where a phase transition or local lattice reconstruction is induced followed by rapid cooling that stabilizes desired material's properties fast enough before the target is altered or damaged by, e.g., hydrodynamic expansion. By using a mask, the longitudinal dimension can be large compared to the transverse dimension, allowing the possibility of rapid transverse cooling. We performed HYDRA simulations that calculate peak temperatures for a series of excitation conditions and cooling rates of silicon targets with micro-structured masks and compare these to a simple analytical model. The model gives scaling laws that can guide the design of targets over a wide range of pulsed ion beam parameters.

  14. Detection of target-probe oligonucleotide hybridization using synthetic nanopore resistive pulse sensing.

    PubMed

    Booth, Marsilea Adela; Vogel, Robert; Curran, James M; Harbison, SallyAnn; Travas-Sejdic, Jadranka

    2013-07-15

    Despite the plethora of DNA sensor platforms available, a portable, sensitive, selective and economic sensor able to rival current fluorescence-based techniques would find use in many applications. In this research, probe oligonucleotide-grafted particles are used to detect target DNA in solution through a resistive pulse nanopore detection technique. Using carbodiimide chemistry, functionalized probe DNA strands are attached to carboxylated dextran-based magnetic particles. Subsequent incubation with complementary target DNA yields a change in surface properties as the two DNA strands hybridize. Particle-by-particle analysis with resistive pulse sensing is performed to detect these changes. A variable pressure method allows identification of changes in the surface charge of particles. As proof-of-principle, we demonstrate that target hybridization is selectively detected at micromolar concentrations (nanomoles of target) using resistive pulse sensing, confirmed by fluorescence and phase analysis light scattering as complementary techniques. The advantages, feasibility and limitations of using resistive pulse sensing for sample analysis are discussed. Copyright © 2013 Elsevier B.V. All rights reserved.

  15. Noninvasive pulsed focused ultrasound allows spatiotemporal control of targeted homing for multiple stem cell types in murine skeletal muscle and the magnitude of cell homing can be increased through repeated applications.

    PubMed

    Burks, Scott R; Ziadloo, Ali; Kim, Saejeong J; Nguyen, Ben A; Frank, Joseph A

    2013-11-01

    Stem cells are promising therapeutics for cardiovascular diseases, and i.v. injection is the most desirable route of administration clinically. Subsequent homing of exogenous stem cells to pathological loci is frequently required for therapeutic efficacy and is mediated by chemoattractants (cell adhesion molecules, cytokines, and growth factors). Homing processes are inefficient and depend on short-lived pathological inflammation that limits the window of opportunity for cell injections. Noninvasive pulsed focused ultrasound (pFUS), which emphasizes mechanical ultrasound-tissue interactions, can be precisely targeted in the body and is a promising approach to target and maximize stem cell delivery by stimulating chemoattractant expression in pFUS-treated tissue prior to cell infusions. We demonstrate that pFUS is nondestructive to murine skeletal muscle tissue (no necrosis, hemorrhage, or muscle stem cell activation) and initiates a largely M2-type macrophage response. We also demonstrate that local upregulation of chemoattractants in pFUS-treated skeletal muscle leads to enhance homing, permeability, and retention of human mesenchymal stem cells (MSC) and human endothelial precursor cells (EPC). Furthermore, the magnitude of MSC or EPC homing was increased when pFUS treatments and cell infusions were repeated daily. This study demonstrates that pFUS defines transient "molecular zip codes" of elevated chemoattractants in targeted muscle tissue, which effectively provides spatiotemporal control and tunability of the homing process for multiple stem cell types. pFUS is a clinically translatable modality that may ultimately improve homing efficiency and flexibility of cell therapies for cardiovascular diseases. © AlphaMed Press.

  16. Nanosurgery of cells and chromosomes using near-infrared twelve-femtosecond laser pulses.

    PubMed

    Uchugonova, Aisada; Lessel, Matthias; Nietzsche, Sander; Zeitz, Christian; Jacobs, Karin; Lemke, Cornelius; König, Karsten

    2012-10-01

    ABSTRACT. Laser-assisted surgery based on multiphoton absorption of near-infrared laser light has great potential for high precision surgery at various depths within the cells and tissues. Clinical applications include refractive surgery (fs-LASIK). The non-contact laser method also supports contamination-free cell nanosurgery. In this paper we describe usage of an ultrashort femtosecond laser scanning microscope for sub-100 nm surgery of human cells and metaphase chromosomes. A mode-locked 85 MHz Ti:Sapphire laser with an M-shaped ultrabroad band spectrum (maxima: 770  nm/830  nm) and an in situ pulse duration at the target ranging from 12 fs up to 3 ps was employed. The effects of laser nanoprocessing in cells and chromosomes have been quantified by atomic force microscopy. These studies demonstrate the potential of extreme ultrashort femtosecond laser pulses at low mean milliwatt powers for sub-100 nm surgery of cells and cellular organelles.

  17. Modeling of intense pulsed ion beam heated masked targets for extreme materials characterization

    DOE PAGES

    Barnard, John J.; Schenkel, Thomas

    2017-11-15

    Intense, pulsed ion beams locally heat materials and deliver dense electronic excitations that can induce material modifications and phase transitions. Material properties can potentially be stabilized by rapid quenching. Pulsed ion beams with pulse lengths of order ns have recently become available for materials processing. Here, we optimize mask geometries for local modification of materials by intense ion pulses. The goal is to rapidly excite targets volumetrically to the point where a phase transition or local lattice reconstruction is induced followed by rapid cooling that stabilizes desired material's properties fast enough before the target is altered or damaged by, e.g.,more » hydrodynamic expansion. By using a mask, the longitudinal dimension can be large compared to the transverse dimension, allowing the possibility of rapid transverse cooling. We performed HYDRA simulations that calculate peak temperatures for a series of excitation conditions and cooling rates of silicon targets with micro-structured masks and compare these to a simple analytical model. In conclusion, the model gives scaling laws that can guide the design of targets over a wide range of pulsed ion beam parameters.« less

  18. Plasma block acceleration based upon the interaction between double targets and an ultra-intense linearly polarized laser pulse

    NASA Astrophysics Data System (ADS)

    Xu, Yanxia; Wang, Jiaxiang; Hora, Heinrich; Qi, Xin; Xing, Yifan; Yang, Lei; Zhu, Wenjun

    2018-04-01

    A new scheme of plasma block acceleration based upon the interaction between double targets and an ultra-intense linearly polarized laser pulse with intensity I ˜ 1022 W/cm2 is investigated via two-dimensional particle-in-cell simulations. The targets are composed of a pre-target of low-density aluminium plasma and an overdense main-target of hydrogen plasma. Through intensive parameter optimization, we have observed highly efficient plasma block accelerations with a monochromatic proton beam peaked at GeVs. The underlying mechanism is attributed to the enhancement of the charge separation field due to the properly selected pre-target.

  19. Dynamic model of target charging by short laser pulse interactions

    NASA Astrophysics Data System (ADS)

    Poyé, A.; Dubois, J.-L.; Lubrano-Lavaderci, F.; D'Humières, E.; Bardon, M.; Hulin, S.; Bailly-Grandvaux, M.; Ribolzi, J.; Raffestin, D.; Santos, J. J.; Nicolaï, Ph.; Tikhonchuk, V.

    2015-10-01

    A model providing an accurate estimate of the charge accumulation on the surface of a metallic target irradiated by a high-intensity laser pulse of fs-ps duration is proposed. The model is confirmed by detailed comparisons with specially designed experiments. Such a model is useful for understanding the electromagnetic pulse emission and the quasistatic magnetic field generation in laser-plasma interaction experiments.

  20. Dynamic model of target charging by short laser pulse interactions.

    PubMed

    Poyé, A; Dubois, J-L; Lubrano-Lavaderci, F; D'Humières, E; Bardon, M; Hulin, S; Bailly-Grandvaux, M; Ribolzi, J; Raffestin, D; Santos, J J; Nicolaï, Ph; Tikhonchuk, V

    2015-10-01

    A model providing an accurate estimate of the charge accumulation on the surface of a metallic target irradiated by a high-intensity laser pulse of fs-ps duration is proposed. The model is confirmed by detailed comparisons with specially designed experiments. Such a model is useful for understanding the electromagnetic pulse emission and the quasistatic magnetic field generation in laser-plasma interaction experiments.

  1. Enhanced Induction of T Cell Immunity Using Dendritic Cells Pulsed with HIV Tat and HCMV-pp65 Fusion Protein In Vitro

    PubMed Central

    Park, Jung-Sun; Park, Soo-Young; Cho, Hyun-Il; Sohn, Hyun-Jung

    2011-01-01

    Background Cytotoxic T lymphocytes (CTLs) appear to play an important role in the control and prevention of human cytomegalovirus (HCMV) infection. The pp65 antigen is a structural protein, which has been defined as a potential target for effective immunity against HCMV infection. Incorporation of an 11 amino acid region of the HIV TAT protein transduction domain (Tat) into protein facilitates rapid, efficient entry into cells. Methods To establish a strategy for the generation of HCMV-specific CTLs in vitro, recombinant truncated N- and C-terminal pp65 protein (pp65 N&C) and N- and C-terminal pp65 protein fused with Tat (Tat/pp65 N&C) was produced in E.coli system. Peripheral blood mononuclear cells were stimulated with dendritic cells (DCs) pulsed with pp65 N&C or Tat/pp65 N&C protein and immune responses induced was examined using IFN-γ ELISPOT assay, cytotoxicity assay and tetramer staining. Results DCs pulsed with Tat/pp65N&C protein could induce higher T-cell responses in vitro compared with pp65N&C. Moreover, the DCs pulsed with Tat/pp65 N&C could stimulate both of CD8+ and CD4+ T-cell responses. The T cells induced by DCs pulsed with Tat/pp65 N&C showed higher cytotoxicity than that of pp65-pulsed DCs against autologous lymphoblastoid B-cell line (LCL) expressing the HCMV-pp65 antigen. Conclusion Our results suggest that DCs pulsed with Tat/pp65 N&C protein effectively induced pp65-specific CTL in vitro. Tat fusion recombinant protein may be useful for the development of adoptive T-cell immunotherapy and DC-based vaccines. PMID:21860612

  2. DNA Damage in Bone Marrow Cells Induced by Femtosecond and Nanosecond Ultraviolet Laser Pulses.

    PubMed

    Morkunas, Vaidotas; Gabryte, Egle; Vengris, Mikas; Danielius, Romualdas; Danieliene, Egle; Ruksenas, Osvaldas

    2015-12-01

    The purpose of this study was to investigate the possible genotoxic impact of new generation 205 nm femtosecond solid-state laser irradiation on the DNA of murine bone marrow cells in vitro, and to compare the DNA damage caused by both femtosecond and nanosecond UV laser pulses. Recent experiments of corneal stromal ablation in vitro and in vivo applying femtosecond UV pulses showed results comparable with or superior to those obtained using nanosecond UV lasers. However, the possible genotoxic effect of ultrashort laser pulses was not investigated. Mouse bone marrow cells were exposed to different doses of 205 nm femtosecond, 213 and 266 nm nanosecond lasers, and 254 nm UV lamp irradiation. The comet assay was used for the evaluation of DNA damage. All types of irradiation demonstrated intensity-dependent genotoxic impact. The DNA damage induced depended mainly upon wavelength rather than on other parameters such as pulse duration, repetition rate, or beam delivery to a target. Both 205 nm femtosecond and clinically applied 213 nm nanosecond lasers' pulses induced a comparable amount of DNA breakage in cells exposed to the same irradiation dose. To further evaluate the suitability of femtosecond UV laser sources for microsurgery, a separate investigation of the genotoxic and mutagenic effects on corneal cells in vitro and, particularly, in vivo is needed.

  3. Pulsed Irradiation Improves Target Selectivity of Infrared Laser-Evoked Gene Operator for Single-Cell Gene Induction in the Nematode C. elegans

    PubMed Central

    Suzuki, Motoshi; Toyoda, Naoya; Takagi, Shin

    2014-01-01

    Methods for turning on/off gene expression at the experimenter’s discretion would be useful for various biological studies. Recently, we reported on a novel microscope system utilizing an infrared laser-evoked gene operator (IR-LEGO) designed for inducing heat shock response efficiently in targeted single cells in living organisms without cell damage, thereby driving expression of a transgene under the control of a heat shock promoter. Although the original IR-LEGO can be successfully used for gene induction, several limitations hinder its wider application. Here, using the nematode Caenorhabditis elegans (C. elegans) as a subject, we have made improvements in IR-LEGO. For better spatial control of heating, a pulsed irradiation method using an optical chopper was introduced. As a result, single cells of C. elegans embryos as early as the 2-cell stage and single neurons in ganglia can be induced to express genes selectively. In addition, the introduction of site-specific recombination systems to IR-LEGO enables the induction of gene expression controlled by constitutive and cell type-specific promoters. The strategies adopted here will be useful for future applications of IR-LEGO to other organisms. PMID:24465705

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

    NASA Astrophysics Data System (ADS)

    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.

  5. Influence of pulsed electromagnetic and pulsed vector magnetic potential field on the growth of tumor cells.

    PubMed

    Loja, Tomas; Stehlikova, Olga; Palko, Lukas; Vrba, Kamil; Rampl, Ivan; Klabusay, Martin

    2014-09-01

    Tumor diseases cause 20% of deaths in Europe and they are the second most common cause of death and morbidity after cardiovascular diseases. Thus, tumor cells are target of many therapeutic strategies and tumor research is focused on searching more efficient and specific drugs as well as new therapeutic approaches. One of the areas of tumor research is an issue of external fields. In our work, we tested influence of a pulsed electromagnetic field (PEMF) and a hypothetic field of the pulsed vector magnetic potential (PVMP) on the growth of tumor cells; and further the possible growth inhibition effect of the PVMP. Both unipolar and bipolar PEMF fields of 5 mT and PVMP fields of 0 mT at frequencies of 15 Hz, 125 Hz and 625 Hz were tested on cancer cell lines derived from various types of tumors: CEM/C2 (acute lymphoblastic leukemia), SU-DHL-4 (B-cell lymphoma), COLO-320DM (colorectal adenocarcinoma), MDA-BM-468 (breast adenocarcinoma), and ZR-75-1 (ductal carcinoma). Cell morphology was observed, proliferation activity using WST assay was measured and simultaneous proportion of live, early apoptotic and dead cells was detected using flow cytometry. A PEMF of 125 Hz and 625 Hz for 24 h-48 h increased proliferation activity in the 2 types of cancer cell lines used, i.e. COLO-320DM and ZR-75-1. In contrast, any of employed methods did not confirm a significant inhibitory effect of hypothetic PVMP field on tumor cells.

  6. Approaches to solar cell design for pulsed laser power receivers

    NASA Technical Reports Server (NTRS)

    Jain, Raj K.; Landis, Geoffrey A.

    1993-01-01

    Using a laser to beam power from Earth to a photovoltaic receiver in space could be a technology with applications to many space missions. Extremely high average-power lasers would be required in a wavelength range of 700-1000 nm. However, high-power lasers inherently operate in a pulsed format. Existing solar cells are not well designed to respond to pulsed incident power. To better understand cell response to pulsed illumination at high intensity, the PC-1D finite-element computer model was used to analyze the response of solar cells to continuous and pulsed laser illumination. Over 50 percent efficiency was calculated for both InP and GaAs cells under steady-state illumination near the optimum wavelength. The time-dependent response of a high-efficiency GaAs concentrator cell to a laser pulse was modeled, and the effect of laser intensity, wavelength, and bias point was studied. Three main effects decrease the efficiency of a solar cell under pulsed laser illumination: series resistance, L-C 'ringing' with the output circuit, and current limiting due to the output inductance. The problems can be solved either by changing the pulse shape or designing a solar cell to accept the pulsed input. Cell design possibilities discussed are a high-efficiency, light-trapping silicon cell, and a monolithic, low-inductance GaAs cell.

  7. An apoptosis targeted stimulus with nanosecond pulsed electric fields (nsPEFs) in E4 squamous cell carcinoma.

    PubMed

    Ren, Wei; Beebe, Stephen J

    2011-04-01

    Stimuli directed towards activation of apoptosis mechanisms are an attractive approach to eliminate evasion of apoptosis, a ubiquitous cancer hallmark. In these in vitro studies, kinetics and electric field thresholds for several apoptosis characteristics are defined in E4 squamous carcinoma cells (SCC) exposed to ten 300 ns pulses with increasing electric fields. Cell death was >95% at the highest electric field and coincident with phosphatidylserine externalization, caspase and calpain activation in the presence and absence of cytochrome c release, decreases in Bid and mitochondria membrane potential (Δψm) without apparent changes reactive oxygen species levels or in Bcl2 and Bclxl levels. Bid cleavage was caspase-dependent (55-60%) and calcium-dependent (40-45%). Intracellular calcium as an intrinsic mechanism and extracellular calcium as an extrinsic mechanism were responsible for about 30 and 70% of calcium dependence for Bid cleavage, respectively. The results reveal electric field-mediated cell death induction and progression, activating pro-apoptotic-like mechanisms and affecting plasma membrane and intracellular functions, primarily through extrinsic-like pathways with smaller contributions from intrinsic-like pathways. Nanosecond second pulsed electric fields trigger heterogeneous cell death mechanisms in E4 SCC populations to delete them, with caspase-associated cell death as a predominant, but not an unaccompanied event.

  8. Intense picosecond pulsed electric fields inhibit proliferation and induce apoptosis of HeLa cells.

    PubMed

    Zhang, Min; Xiong, Zheng-Ai; Chen, Wen-Juan; Yao, Cheng-Guo; Zhao, Zhong-Yong; Hua, Yuan-Yuan

    2013-06-01

    A picosecond pulsed electric field (psPEF) is a localized physical therapy for tumors that has been developed in recent years, and that may in the future be utilized as a targeted non‑invasive treatment. However, there are limited studies regarding the biological effects of psPEF on cells. Electric field amplitude and pulse number are the main parameters of psPEF that influence its biological effects. In this study, we exposed HeLa cells to a psPEF with a variety of electric field amplitudes, from 100 to 600 kV/cm, and various pulse numbers, from 1,000 to 3,000. An MTT assay was used to detect the growth inhibition, while flow cytometry was used to determine the occurrence of apoptosis and the cell cycle of the HeLa cells following treatment. The morphological changes during cell apoptosis were observed using transmission electron microscopy (TEM). The results demonstrated that the cell growth inhibition rate gradually increased, in correlation with the increasing electric field amplitude and pulse number, and achieved a plateau of maximum cell inhibition 12 h following the pulses. In addition, typical characteristics of HeLa cell apoptosis in the experimental groups were observed by TEM. The results demonstrated that the rate of apoptosis in the experimental groups was significantly elevated in comparison with the untreated group. In the treatment groups, the rate of apoptosis was greater in the higher amplitude groups than in the lower amplitude groups. The same results were obtained when the variable was the pulse number. Flow cytometric analysis indicated that the cell cycle of the HeLa cells was arrested at the G2/M phase following psPEF treatment. Overall, our results indicated that psPEF inhibited cell proliferation and induced cell apoptosis, and that these effects occurred in a dose-dependent manner. In addition, the results demonstrated that the growth of the HeLa cells was arrested at the G2/M phase following treatment. This study may provide a

  9. A bright attosecond x-ray pulse train generation in a double-laser-driven cone target

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    Hu, Li-Xiang; Yu, Tong-Pu, E-mail: tongpu@nudt.edu.cn; Shao, Fu-Qiu

    By using full three-dimensional particle-in-cell and Monte Carlo simulations, we investigate the generation of a high-brightness attosecond x-ray pulse train in a double-laser-driven cone target. The scheme makes use of two lasers: the first high-intensity laser with a laser peak intensity 1.37 × 10{sup 20 }W/cm{sup 2} irradiates the cone and produces overdense attosecond electron bunches; the second counterpropagating weakly relativistic laser with a laser peak intensity 4.932 × 10{sup 17 }W/cm{sup 2} interacts with the produced electron bunches and a bright x-ray pulse train is generated by Thomson backscattering of the second laser off the attosecond electron bunches. It is shown that the photon fluxmore » rises by 5 times using the cone target as compared with a normal channel. Meanwhile, the x-ray peak brightness increases significantly from 1.4 × 10{sup 21}/(s mm{sup 2} mrad{sup 2} 0.1 keV) to 6.0 × 10{sup 21}/(s mm{sup 2} mrad{sup 2} 0.1 keV), which is much higher than that of the Thomson x-ray source generated from traditional accelerators. We also discuss the influence of the laser and target parameters on the x-ray pulse properties. This compact bright x-ray source may have diverse applications, e.g., the study of electric dynamics and harmonics emission in the atomic scale.« less

  10. Size-based cell sorting with a resistive pulse sensor and an electromagnetic pump in a microfluidic chip.

    PubMed

    Song, Yongxin; Li, Mengqi; Pan, Xinxiang; Wang, Qi; Li, Dongqing

    2015-02-01

    An electrokinetic microfluidic chip is developed to detect and sort target cells by size from human blood samples. Target-cell detection is achieved by a differential resistive pulse sensor (RPS) based on the size difference between the target cell and other cells. Once a target cell is detected, the detected RPS signal will automatically actuate an electromagnetic pump built in a microchannel to push the target cell into a collecting channel. This method was applied to automatically detect and sort A549 cells and T-lymphocytes from a peripheral fingertip blood sample. The viability of A549 cells sorted in the collecting well was verified by Hoechst33342 and propidium iodide staining. The results show that as many as 100 target cells per minute can be sorted out from the sample solution and thus is particularly suitable for sorting very rare target cells, such as circulating tumor cells. The actuation of the electromagnetic valve has no influence on RPS cell detection and the consequent cell-sorting process. The viability of the collected A549 cell is not impacted by the applied electric field when the cell passes the RPS detection area. The device described in this article is simple, automatic, and label-free and has wide applications in size-based rare target cell sorting for medical diagnostics. © 2014 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  11. EFFECTS OF LASER RADIATION ON MATTER: Simulation of photon acceleration upon irradiation of a mylar target by femtosecond laser pulses

    NASA Astrophysics Data System (ADS)

    Andreev, Stepan N.; Rukhadze, Anri A.; Tarakanov, V. P.; Yakutov, B. P.

    2010-01-01

    Acceleration of protons is simulated by the particle-in-cell (PIC) method upon irradiation of mylar targets of different thicknesses by femtosecond plane-polarised pulsed laser radiation and at different angles of radiation incidence on the target. The comparison of the results of calculations with the experimental data obtained in recent experiments shows their good agreement. The optimal angle of incidence (458) at which the proton energy achieves its absolute maximum is obtained.

  12. Plasmonic nanobubbles for target cell-specific gene and drug delivery and multifunctional processing of heterogeneous cell systems

    NASA Astrophysics Data System (ADS)

    Lukianova-Hleb, Ekaterina Y.; Huye, Leslie E.; Brenner, Malcolm K.; Lapotko, Dmitri O.

    2014-03-01

    Cell and gene cancer therapies require ex vivo cell processing of human grafts. Such processing requires at least three steps - cell enrichment, cell separation (destruction), and gene transfer - each of which requires the use of a separate technology. While these technologies may be satisfactory for research use, they are of limited usefulness in the clinical treatment setting because they have a low processing rate, as well as a low transfection and separation efficacy and specificity in heterogeneous human grafts. Most problematic, because current technologies are administered in multiple steps - rather than in a single, multifunctional, and simultaneous procedure - they lengthen treatment process and introduce an unnecessary level of complexity, labor, and resources into clinical treatment; all these limitations result in high losses of valuable cells. We report a universal, high-throughput, and multifunctional technology that simultaneously (1) inject free external cargo in target cells, (2) destroys unwanted cells, and (3) preserve valuable non-target cells in heterogeneous grafts. Each of these functions has single target cell specificity in heterogeneous cell system, processing rate > 45 mln cell/min, injection efficacy 90% under 96% viability of the injected cells, target cell destruction efficacy > 99%, viability of not-target cells >99% The developed technology employs novel cellular agents, called plasmonic nanobubbles (PNBs). PNBs are not particles, but transient, intracellular events, a vapor nanobubbles that expand and collapse in mere nanoseconds under optical excitation of gold nanoparticles with short picosecond laser pulses. PNBs of different, cell-specific, size (1) inject free external cargo with small PNBs, (2) Destroy other target cells mechanically with large PNBs and (3) Preserve non-target cells. The multi-functionality, precision, and high throughput of all-in-one PNB technology will tremendously impact cell and gene therapies and other

  13. Granule size control and targeting in pulsed spray fluid bed granulation.

    PubMed

    Ehlers, Henrik; Liu, Anchang; Räikkönen, Heikki; Hatara, Juha; Antikainen, Osmo; Airaksinen, Sari; Heinämäki, Jyrki; Lou, Honxiang; Yliruusi, Jouko

    2009-07-30

    The primary aim of the study was to investigate the effects of pulsed liquid feed on granule size. The secondary aim was to increase knowledge of this technique in granule size targeting. Pulsed liquid feed refers to the pump changing between on- and off-positions in sequences, called duty cycles. One duty cycle consists of one on- and off-period. The study was performed with a laboratory-scale top-spray fluid bed granulator with duty cycle length and atomization pressure as studied variables. The liquid feed rate, amount and inlet air temperature were constant. The granules were small, indicating that the powder has only undergone ordered mixing, nucleation and early growth. The effect of atomizing pressure on granule size depends on inlet air relative humidity, with premature binder evaporation as a reason. The duty cycle length was of critical importance to the end product attributes, by defining the extent of intermittent drying and rewetting. By varying only the duty cycle length, it was possible to control granule nucleation and growth, with a wider granule size target range in increased relative humidity. The present study confirms that pulsed liquid feed in fluid bed granulation is a useful tool in end product particle size targeting.

  14. Intense picosecond pulsed electric fields induce apoptosis through a mitochondrial-mediated pathway in HeLa cells.

    PubMed

    Hua, Yuan-Yuan; Wang, Xiao-Shu; Zhang, Yu; Yao, Chen-Guo; Zhang, Xi-Ming; Xiong, Zheng-Ai

    2012-04-01

    The application of pulsed electric fields (PEF) is emerging as a new technique for tumor therapy. Picosecond pulsed electric fields (psPEF) can be transferred to target deep tissue non-invasively and precisely, but the research of the biological effects of psPEF on cells is limited. Electric theory predicts that intense psPEF will target mitochondria and lead to changes in transmembrane potential, therefore, it is hypothesized that it can induce mitochondrial-mediated apoptosis. HeLa cells were exposed to psPEF in this study to investigate this hypothesis. MTT assay demonstrated that intense psPEF significantly inhibited the proliferation of HeLa cells in a dose-dependent manner. Typical characteristics of apoptosis in HeLa cells were observed, using transmission electron microscopy. Loss of mitochondrial transmembrane potential was explored using laser scanning confocal microscopy with Rhodamine-123 (Rh123) staining. Furthermore, the mitochondrial apoptotic events were also confirmed by western blot analysis for the release of cytochrome C and apoptosis-inducing factor from mitochondria into the cytosol. In addition, activation of caspase-3, caspase-9, upregulation of Bax, p53 and downregulation of Bcl-2 were observed in HeLa cells also indicating apoptosis. Taken together, these results demonstrate that intense psPEF induce cell apoptosis through a mitochondrial-mediated pathway.

  15. Preparation of Ga-doped ZnO films by pulsed dc magnetron sputtering with cylindrical rotating target for thin film solar cell applications

    NASA Astrophysics Data System (ADS)

    Shin, Beom-Ki; Lee, Tae-Il; Park, Ji-Hyeon; Park, Kang-Il; Ahn, Kyung-Jun; Park, Sung-Kee; Lee, Woong; Myoung, Jae-Min

    2011-11-01

    Applicability of Ga-doped ZnO (GZO) films for thin film solar cells (TFSCs) was investigated by preparing GZO films via pulsed dc magnetron sputtering (PDMS) with rotating target. The GZO films showed improved crystallinity and increasing degree of Ga doping with increasing thickness to a limit of 1000 nm. The films also fulfilled requirements for the transparent electrodes of TFSCs in terms of electrical and optical properties. Moreover, the films exhibited good texturing potential based on etching studies with diluted HCl, which yielded an improved light trapping capability without significant degradation in electrical propreties. It is therefore suggested that the surface-textured GZO films prepared via PDMS and etching are promising candidates for indium-free transparent electrodes for TFSCs.

  16. Enhanced cytotoxic activity of effector T-cells against cholangiocarcinoma by dendritic cells pulsed with pooled mRNA.

    PubMed

    Junking, Mutita; Grainok, Janya; Thepmalee, Chutamas; Wongkham, Sopit; Yenchitsomanus, Pa-Thai

    2017-10-01

    Cholangiocarcinoma is a malignancy of bile duct epithelia with an increasing in incidence rate worldwide. Surgery is the only curative treatment, while adjuvant chemotherapy and radiotherapy render poor responses. Cell-based immunotherapy is a potential strategy for cholangiocarcinoma treatment. However, variation of tumor antigens in cholangiocarcinoma leads to the ineffectiveness of cell-based immunotherapy. In this study, we examined the activation of effector T-cells by dendritic cells pulsed with protein lysate or total RNA from cholangiocarcinoma cell lines for their cytolytic activity against cholangiocarcinoma. Broad-spectrum antigen types with respect to RNA antigen sources were obtained from combination of three cholangiocarcinoma cell lines (KKU-213, KKU-100, and KKU-055). Compared with protein lysate-pulsed dendritic cells, total RNA-pulsed dendritic cells induced anti-tumor effector T-cell response with higher killing ability to KKU-100 and KKU-213 cells compared with protein lysate-pulsed dendritic cells. Moreover, pooled messenger RNA from three cholangiocarcinoma cell lines significantly increased the specific killing capacity of activated lymphocytes against KKU-213 cells. These results suggest that activation of anti-tumor effector T-cells against cholangiocarcinoma by RNA-pulsed dendritic cells is more effective than that by protein lysate-pulsed dendritic cells. In addition, pulsing dendritic cells with pooled messenger RNA from multiple cell lines enhanced the efficacy of a cellular immune response against cholangiocarcinoma.

  17. Nanoshell-mediated targeted photothermal therapy of HER2 human breast cancer cells using pulsed and continuous wave lasers: an in vitro study.

    PubMed

    Khosroshahi, Mohammad E; Hassannejad, Zahra; Firouzi, Masoumeh; Arshi, Ahmad R

    2015-09-01

    In this study, we report the apoptosis induction in HER2 overexpressed breast cancer cells using pulsed, continuous wave lasers and polyvinylpyrrolidone (PVP)-stabilized magneto-plasmonic nanoshells (PVP-MPNS) delivered by immunoliposomes. The immunoliposomes containing PVP-MPNS were fabricated and characterized. Heating efficiency of the synthesized nanostructures was calculated. The effect of functionalization on cellular uptake of nanoparticles was assessed using two cell lines of BT-474 and Calu-6. The best uptake result was achieved by functionalized liposome (MPNS-LAb) and BT-474. Also, the interaction of 514 nm argon (Ar) and Nd/YAG second harmonic 532-nm lasers with nanoparticles was investigated based on the temperature rise of the nanoshell suspension and the release value of 5(6)-carboxyfluorescein (CF) from CF/MPNS-loaded liposomes. The temperature increase of the suspensions after ten consecutive pulses of 532 nm and 5 min of irradiation by Ar laser were measured approximately 2 and 12 °C, respectively. The irradiation of CF/MPNS-loaded liposomes by Ar laser for 3 min resulted in 24.3 % release of CF, and in the case of 532 nm laser, the release was laser energy dependent. Furthermore, the comparison of CF release showed a higher efficiency for the Ar laser than by direct heating of nanoshell suspension using circulating water. The percentage of cell apoptosis after irradiation by Ar and 532 nm lasers were 44.6 and 42.6 %, respectively. The obtained results suggest that controlling the NP-laser interaction using optical properties of nanoshells and the laser parameters can be used to develop a new cancer therapy modality via targeted nanoshell and drug delivery.

  18. Nanopore formation in neuroblastoma cells following ultrashort electric pulse exposure

    NASA Astrophysics Data System (ADS)

    Roth, Caleb C.; Payne, Jason A.; Wilmink, Gerald J.; Ibey, Bennett L.

    2011-03-01

    Ultrashort or nanosecond electrical pulses (USEP) cause repairable damage to the plasma membranes of cells through formation of nanopores. These nanopores are able to pass small ions such as sodium, calcium, and potassium, but remain impermeable to larger molecules like trypan blue and propidium iodide. What remains uncertain is whether generation of nanopores by ultrashort electrical pulses can inhibit action potentials in excitable cells. In this paper, we explored the sensitivity of excitable cells to USEP using Calcium Green AM 1 ester fluorescence to measure calcium uptake indicative of nanopore formation in the plasma membrane. We determined the threshold for nanopore formation in neuroblastoma cells for three pulse parameters (amplitude, pulse width, and pulse number). Measurement of such thresholds will guide future studies to determine if USEP can inhibit action potentials without causing irreversible membrane damage.

  19. Multifunctional gold nanorods for selective plasmonic photothermal therapy in pancreatic cancer cells using ultra-short pulse near-infrared laser irradiation.

    PubMed

    Patino, Tania; Mahajan, Ujjwal; Palankar, Raghavendra; Medvedev, Nikolay; Walowski, Jakob; Münzenberg, Markus; Mayerle, Julia; Delcea, Mihaela

    2015-03-12

    Gold nanorods (AuNRs) have attracted considerable attention in plasmonic photothermal therapy for cancer treatment by exploiting their selective and localized heating effect due to their unique photophysical properties. Here we describe a strategy to design a novel multifunctional platform based on AuNRs to: (i) specifically target the adenocarcinoma MUC-1 marker through the use of the EPPT-1 peptide, (ii) enhance cellular uptake through a myristoylated polyarginine peptide (MPAP) and (iii) selectively induce cell death by ultra-short near infrared laser pulses. We used a biotin-avidin based approach to conjugate EPPT-1 and MPAP to AuNRs. Dual-peptide (EPPT-1+MPAP) labelled AuNRs showed a significantly higher uptake by pancreatic ductal adenocarcinoma cells when compared to their single peptide or avidin conjugated counterparts. In addition, we selectively induced cell death by ultra-short near infrared laser pulses in small target volumes (∼1 μm3), through the creation of plasmonic nanobubbles that lead to the destruction of a local cell environment. Our approach opens new avenues for conjugation of multiple ligands on AuNRs targeting cancer cells and tumors and it is relevant for plasmonic photothermal therapy.

  20. The efficiency of photovoltaic cells exposed to pulsed laser light

    NASA Technical Reports Server (NTRS)

    Lowe, R. A.; Landis, G. A.; Jenkins, P.

    1993-01-01

    Future space missions may use laser power beaming systems with a free electron laser (FEL) to transmit light to a photovoltaic array receiver. To investigate the efficiency of solar cells with pulsed laser light, several types of GaAs, Si, CuInSe2, and GaSb cells were tested with the simulated pulse format of the induction and radio frequency (RF) FEL. The induction pulse format was simulated with an 800-watt average power copper vapor laser and the RF format with a frequency-doubled mode-locked Nd:YAG laser. Averaged current vs bias voltage measurements for each cell were taken at various optical power levels and the efficiency measured at the maximum power point. Experimental results show that the conversion efficiency for the cells tested is highly dependent on cell minority carrier lifetime, the width and frequency of the pulses, load impedance, and the average incident power. Three main effects were found to decrease the efficiency of solar cells exposed to simulated FEL illumination: cell series resistance, LC 'ringing', and output inductance. Improvements in efficiency were achieved by modifying the frequency response of the cell to match the spectral energy content of the laser pulse with external passive components.

  1. Investigation of longitudinal proton acceleration in exploded targets irradiated by intense short-pulse laser

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    Gauthier, M.; CEA, DAM, DIF, 91297 Arpajon; Lévy, A.

    2014-01-15

    It was recently shown that a promising way to accelerate protons in the forward direction to high energies is to use under-dense or near-critical density targets instead of solids. Simulations have revealed that the acceleration process depends on the density gradients of the plasma target. Indeed, under certain conditions, the most energetic protons are predicted to be accelerated by a collisionless shock mechanism that significantly increases their energy. We report here the results of a recent experiment dedicated to the study of longitudinal ion acceleration in partially exploded foils using a high intensity (∼5 × 10{sup 18} W/cm{sup 2}) picosecond laser pulse. Wemore » show that protons accelerated using targets having moderate front and rear plasma gradients (up to ∼8 μm gradient length) exhibit similar maximum proton energy and number compared to proton beams that are produced, in similar laser conditions, from solid targets, in the well-known target normal sheath acceleration regime. Particle-In-Cell simulations, performed in the same conditions as the experiment and consistent with the measurements, allow laying a path for further improvement of this acceleration scheme.« less

  2. Multifunctional gold nanorods for selective plasmonic photothermal therapy in pancreatic cancer cells using ultra-short pulse near-infrared laser irradiation

    NASA Astrophysics Data System (ADS)

    Patino, Tania; Mahajan, Ujjwal; Palankar, Raghavendra; Medvedev, Nikolay; Walowski, Jakob; Münzenberg, Markus; Mayerle, Julia; Delcea, Mihaela

    2015-03-01

    Gold nanorods (AuNRs) have attracted considerable attention in plasmonic photothermal therapy for cancer treatment by exploiting their selective and localized heating effect due to their unique photophysical properties. Here we describe a strategy to design a novel multifunctional platform based on AuNRs to: (i) specifically target the adenocarcinoma MUC-1 marker through the use of the EPPT-1 peptide, (ii) enhance cellular uptake through a myristoylated polyarginine peptide (MPAP) and (iii) selectively induce cell death by ultra-short near infrared laser pulses. We used a biotin-avidin based approach to conjugate EPPT-1 and MPAP to AuNRs. Dual-peptide (EPPT-1 + MPAP) labelled AuNRs showed a significantly higher uptake by pancreatic ductal adenocarcinoma cells when compared to their single peptide or avidin conjugated counterparts. In addition, we selectively induced cell death by ultra-short near infrared laser pulses in small target volumes (~1 μm3), through the creation of plasmonic nanobubbles that lead to the destruction of a local cell environment. Our approach opens new avenues for conjugation of multiple ligands on AuNRs targeting cancer cells and tumors and it is relevant for plasmonic photothermal therapy.Gold nanorods (AuNRs) have attracted considerable attention in plasmonic photothermal therapy for cancer treatment by exploiting their selective and localized heating effect due to their unique photophysical properties. Here we describe a strategy to design a novel multifunctional platform based on AuNRs to: (i) specifically target the adenocarcinoma MUC-1 marker through the use of the EPPT-1 peptide, (ii) enhance cellular uptake through a myristoylated polyarginine peptide (MPAP) and (iii) selectively induce cell death by ultra-short near infrared laser pulses. We used a biotin-avidin based approach to conjugate EPPT-1 and MPAP to AuNRs. Dual-peptide (EPPT-1 + MPAP) labelled AuNRs showed a significantly higher uptake by pancreatic ductal adenocarcinoma

  3. Hydrodynamic Determinants of Cell Necrosis and Molecular Delivery Produced by Pulsed Laser Microbeam Irradiation of Adherent Cells

    PubMed Central

    Compton, Jonathan L.; Hellman, Amy N.; Venugopalan, Vasan

    2013-01-01

    Time-resolved imaging, fluorescence microscopy, and hydrodynamic modeling were used to examine cell lysis and molecular delivery produced by picosecond and nanosecond pulsed laser microbeam irradiation in adherent cell cultures. Pulsed laser microbeam radiation at λ = 532 nm was delivered to confluent monolayers of PtK2 cells via a 40×, 0.8 NA microscope objective. Using laser microbeam pulse durations of 180–1100 ps and pulse energies of 0.5–10.5 μJ, we examined the resulting plasma formation and cavitation bubble dynamics that lead to laser-induced cell lysis, necrosis, and molecular delivery. The cavitation bubble dynamics are imaged at times of 0.5 ns to 50 μs after the pulsed laser microbeam irradiation, and fluorescence assays assess the resulting cell viability and molecular delivery of 3 kDa dextran molecules. Reductions in both the threshold laser microbeam pulse energy for plasma formation and the cavitation bubble energy are observed with decreasing pulse duration. These energy reductions provide for increased precision of laser-based cellular manipulation including cell lysis, cell necrosis, and molecular delivery. Hydrodynamic analysis reveals critical values for the shear-stress impulse generated by the cavitation bubble dynamics governs the location and spatial extent of cell necrosis and molecular delivery independent of pulse duration and pulse energy. Specifically, cellular exposure to a shear-stress impulse J≳0.1 Pa s ensures cell lysis or necrosis, whereas exposures in the range of 0.035≲J≲0.1 Pa s preserve cell viability while also enabling molecular delivery of 3 kDa dextran. Exposure to shear-stress impulses of J≲0.035 Pa s leaves the cells unaffected. Hydrodynamic analysis of these data, combined with data from studies of 6 ns microbeam irradiation, demonstrates the primacy of shear-stress impulse in determining cellular outcome resulting from pulsed laser microbeam irradiation spanning a nearly two

  4. Sacrificial-layer free transfer of mammalian cells using near infrared femtosecond laser pulses

    PubMed Central

    Zhang, Jun; Hartmann, Bastian; Siegel, Julian; Marchi, Gabriele; Clausen-Schaumann, Hauke; Sudhop, Stefanie; Huber, Heinz P.

    2018-01-01

    Laser-induced cell transfer has been developed in recent years for the flexible and gentle printing of cells. Because of the high transfer rates and the superior cell survival rates, this technique has great potential for tissue engineering applications. However, the fact that material from an inorganic sacrificial layer, which is required for laser energy absorption, is usually transferred to the printed target structure, constitutes a major drawback of laser based cell printing. Therefore alternative approaches using deep UV laser sources and protein based acceptor films for energy absorption, have been introduced. Nevertheless, deep UV radiation can introduce DNA double strand breaks, thereby imposing the risk of carcinogenesis. Here we present a method for the laser-induced transfer of hydrogels and mammalian cells, which neither requires any sacrificial material for energy absorption, nor the use of UV lasers. Instead, we focus a near infrared femtosecond (fs) laser pulse (λ = 1030 nm, 450 fs) directly underneath a thin cell layer, suspended on top of a hydrogel reservoir, to induce a rapidly expanding cavitation bubble in the gel, which generates a jet of material, transferring cells and hydrogel from the gel/cell reservoir to an acceptor stage. By controlling laser pulse energy, well-defined cell-laden droplets can be transferred with high spatial resolution. The transferred human (SCP1) and murine (B16F1) cells show high survival rates, and good cell viability. Time laps microscopy reveals unaffected cell behavior including normal cell proliferation. PMID:29718923

  5. Plasmonic nanodiamonds: targeted core-shell type nanoparticles for cancer cell thermoablation.

    PubMed

    Rehor, Ivan; Lee, Karin L; Chen, Kevin; Hajek, Miroslav; Havlik, Jan; Lokajova, Jana; Masat, Milan; Slegerova, Jitka; Shukla, Sourabh; Heidari, Hamed; Bals, Sara; Steinmetz, Nicole F; Cigler, Petr

    2015-02-18

    Targeted biocompatible nanostructures with controlled plasmonic and morphological parameters are promising materials for cancer treatment based on selective thermal ablation of cells. Here, core-shell plasmonic nanodiamonds consisting of a silica-encapsulated diamond nanocrystal coated in a gold shell are designed and synthesized. The architecture of particles is analyzed and confirmed in detail using electron tomography. The particles are biocompatibilized using a PEG polymer terminated with bioorthogonally reactive alkyne groups. Azide-modified transferrin is attached to these particles, and their high colloidal stability and successful targeting to cancer cells overexpressing the transferrin receptor are demonstrated. The particles are nontoxic to the cells and they are readily internalized upon binding to the transferrin receptor. The high plasmonic cross section of the particles in the near-infrared region is utilized to quantitatively ablate the cancer cells with a short, one-minute irradiation by a pulse 750-nm laser. © 2014 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  6. Pulsed DC Electric Field–Induced Differentiation of Cortical Neural Precursor Cells

    PubMed Central

    Chang, Hui-Fang; Lee, Ying-Shan; Tang, Tang K.; Cheng, Ji-Yen

    2016-01-01

    We report the differentiation of neural stem and progenitor cells solely induced by direct current (DC) pulses stimulation. Neural stem and progenitor cells in the adult mammalian brain are promising candidates for the development of therapeutic neuroregeneration strategies. The differentiation of neural stem and progenitor cells depends on various in vivo environmental factors, such as nerve growth factor and endogenous EF. In this study, we demonstrated that the morphologic and phenotypic changes of mouse neural stem and progenitor cells (mNPCs) could be induced solely by exposure to square-wave DC pulses (magnitude 300 mV/mm at frequency of 100-Hz). The DC pulse stimulation was conducted for 48 h, and the morphologic changes of mNPCs were monitored continuously. The length of primary processes and the amount of branching significantly increased after stimulation by DC pulses for 48 h. After DC pulse treatment, the mNPCs differentiated into neurons, astrocytes, and oligodendrocytes simultaneously in stem cell maintenance medium. Our results suggest that simple DC pulse treatment could control the fate of NPCs. With further studies, DC pulses may be applied to manipulate NPC differentiation and may be used for the development of therapeutic strategies that employ NPCs to treat nervous system disorders. PMID:27352251

  7. Thermonuclear targets for direct-drive ignition by a megajoule laser pulse

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    Bel’kov, S. A.; Bondarenko, S. V.; Vergunova, G. A.

    2015-10-15

    Central ignition of a thin two-layer-shell fusion target that is directly driven by a 2-MJ profiled pulse of Nd laser second-harmonic radiation has been studied. The parameters of the target were selected so as to provide effective acceleration of the shell toward the center, which was sufficient for the onset of ignition under conditions of increased hydrodynamic stability of the ablator acceleration and compression. The aspect ratio of the inner deuterium-tritium layer of the shell does not exceed 15, provided that a major part (above 75%) of the outer layer (plastic ablator) is evaporated by the instant of maximum compression.more » The investigation is based on two series of numerical calculations that were performed using one-dimensional (1D) hydrodynamic codes. The first 1D code was used to calculate the absorption of the profiled laser-radiation pulse (including calculation of the total absorption coefficient with allowance for the inverse bremsstrahlung and resonance mechanisms) and the spatial distribution of target heating for a real geometry of irradiation using 192 laser beams in a scheme of focusing with a cubo-octahedral symmetry. The second 1D code was used for simulating the total cycle of target evolution under the action of absorbed laser radiation and for determining the thermonuclear gain that was achieved with a given target.« less

  8. Optoacoustic imaging of gold nanoparticles targeted to breast cancer cells

    NASA Astrophysics Data System (ADS)

    Eghtedari, Mohammad; Motamedi, Massoud; Popov, Vsevolod L.; Kotov, Nicholas A.; Oraevsky, Alexander A.

    2004-07-01

    Optoacoustic Tomography (OAT) is a rapidly growing technology that enables noninvasive deep imaging of biological tissues based on their light absorption. In OAT, the interaction of a pulsed laser with tissue increases the temperature of the absorbing components in a confined volume of tissue. Rapid perturbation of the temperature (<1°C) deep within tissue produces weak acoustic waves that easily travel to the surface of the tissue with minor attenuation. Abnormal angiogenesis in a malignant tumor, that increases its blood content, makes a native contrast for optoacoustic imaging; however, the application of OAT for early detection of malignant tumors requires the enhancement of optoacoustic signals originated from tumor by using an exogenous contrast agent. Due to their strong absorption, we have used gold nanoparticles (NP) as a contrast agent. 40nm spherical gold nanoparticles were attached to monoclonal antibody to target cell surface of breast cancer cells. The targeted cancer cells were implanted at depth of 5-6cm within a gelatinous object that optically resembles human breast. Experimental sensitivity measurements along with theoretical analysis showed that our optoacoustic imaging system is capable of detecting a phantom breast tumor with the volume of 0.15ml, which is composed of 25 million NP-targeted cancer cells, at a depth of 5 centimeters in vitro.

  9. Evidence for speckle effects on pulsed CO2 lidar signal returns from remote targets

    NASA Technical Reports Server (NTRS)

    Menzies, R. T.; Kavaya, M. J.; Flamant, P. H.

    1984-01-01

    A pulsed CO2 lidar was used to study statistical properties of signal returns from various rough surfaces at distances near 2 km. These included natural in situ topographic materials as well as man-made hard targets. Three lidar configurations were used: heterodyne detection with single temporal mode transmitter pulses, and direct detection with single and multiple temporal mode pulses. The significant differences in signal return statistics, due largely to speckle effects, are discussed.

  10. Theoretical studies of defect formation and target heating by intense pulsed ion beams

    NASA Astrophysics Data System (ADS)

    Barnard, J. J.; Schenkel, T.; Persaud, A.; Seidl, P. A.; Friedman, A.; Grote, D. P.; Davidson, R. C.; Gilson, E. P.; Kaganovich, I.

    2015-11-01

    We present results of three studies related to experiments on NDCX-II, the Neutralized Drift Compression Experiment, a short-pulse (~ 1ns), high-current (~ 70A) linear accelerator for 1.2 MeV ions at LBNL. These include: (a) Coupled transverse and longitudinal envelope calculations of the final non-neutral ion beam transport, followed by neutralized drift and final focus, for a number of focus and drift lengths and with a series of ion species (Z =1-19). Predicted target fluences were obtained and target temperatures in the 1 eV range estimated. (b) HYDRA simulations of the target response for Li and He ions and for Al and Au targets at various ion fluences (up to 1012 ions/pulse/mm2) and pulse durations, benchmarking temperature estimates from the envelope calculations. (c) Crystal-Trim simulations of ion channeling through single-crystal lattices, with comparisons to ion transmission data as a function of orientation angle of the crystal foil and for different ion intensities and ion species. This work was performed under the auspices of the U.S. DOE under contracts DE-AC52-07NA27344 (LLNL), DE-AC02-05CH11231 (LBNL) and DE-AC02-76CH0307 (PPPL) and was supported by the US DOE Office of Science, Fusion Energy Sciences. LLNL-ABS-67521.

  11. On-target diagnosing of few-cycle pulses by high-order-harmonic generation

    NASA Astrophysics Data System (ADS)

    Brambila, Danilo S.; Husakou, Anton; Ivanov, Misha; Zhavoronkov, Nickolai

    2017-12-01

    We propose an approach to determine the residual phase distortion directly in the interaction region of few-cycle laser radiation with a gaseous target. We describe how the spectra of the generated high harmonics measured as a function of externally introduced dispersion into the driving few-cycle laser pulse can be used to decode small amounts of second- and third-order spectral phase, including the sign. The diagnosis is based on the analysis of several key features in the high-harmonic spectrum: the depth of spectral modulation, the position of the cutoff, and the symmetry of the spectrum with respect to the introduced dispersion. The approach is applicable to pulses without carrier-envelope phase (CEP) stabilization. Surprisingly, we find that for nearly-single-cycle pulses with nonstabilized CEP, deep spectral modulations in the harmonic spectra emerge for positively rather than negatively chirped pulses, in contrast to the case of CEP-stabilized pulses.

  12. Microjet formation and hard x-ray production from a liquid metal target irradiated by intense femtosecond laser pulses

    NASA Astrophysics Data System (ADS)

    Lar'kin, A.; Uryupina, D.; Ivanov, K.; Savel'ev, A.; Bonnet, T.; Gobet, F.; Hannachi, F.; Tarisien, M.; Versteegen, M.; Spohr, K.; Breil, J.; Chimier, B.; Dorchies, F.; Fourment, C.; Leguay, P.-M.; Tikhonchuk, V. T.

    2014-09-01

    By using a liquid metal as a target one may significantly enhance the yield of hard x-rays with a sequence of two intense femtosecond laser pulses. The influence of the time delay between the two pulses is studied experimentally and interpreted with numerical simulations. It was suggested that the first arbitrary weak pulse produces microjets from the target surface, while the second intense pulse provides an efficient electron heating and acceleration along the jet surface. These energetic electrons are the source of x-ray emission while striking the target surface. The microjet formation is explained based on the results given by both optical diagnostics and hydrodynamic modeling by a collision of shocks originated from two distinct zones of laser energy deposition.

  13. Basic features of electromagnetic pulse generated in a laser-target chamber at 3-TW laser facility PALS

    NASA Astrophysics Data System (ADS)

    De Marco, M.; Pfeifer, M.; Krousky, E.; Krasa, J.; Cikhardt, J.; Klir, D.; Nassisi, V.

    2014-04-01

    We describe the radiofrequency emission taking place when 300 ps laser pulses irradiate various solid targets with an intensity of 1016 W/cm2. The emission of intense electromagnetic pulses was observed outside the laser target chamber by two loop antennas up to 1 GHz. Electromagnetic pulses can be 800 MHz transients, which decay from a peak electromagnetic field of E0 ≊ 7 kV/m and H0 ≊ 15 A/m. The occurrence of these electromagnetic pulses is associated with generation of hard x-rays with photon energies extending beyond 1 MeV. This contribution reports the first observation of this effect at the PALS facility.

  14. Thermal-hydraulic simulation of mercury target concepts for a pulsed spallation neutron source

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    Siman-Tov, M.; Wendel, M.; Haines, J.

    1996-06-01

    The Oak Ridge Spallation Neutron Source (ORSNS) is a high-power, accelerator-based pulsed spallation neutron source being designed by a multi-laboratory team led by Oak Ridge National Laboratory to achieve very high fluxes of neutrons for scientific experiments. The ORSNS is projected to have a 1 MW proton beam upgradable to 5 MW. About 60% of the beam power (1-5 MW, 17-83 kJ/pulse in 0.5 microsec at 60 cps) is deposited in the liquid metal (mercury) target having the dimensions of 65x30x10 cm (about 19.5 liter). Peak steady state power density is about 150 and 785 MW/m{sup 3} for 1 MWmore » and 5 MW beam respectively, whereas peak pulsed power density is as high as 5.2 and 26.1 GW/m{sup 3}, respectively. The peak pulse temperature rise rate is 14 million C/s (for 5 MW beam) whereas the total pulse temperature rise is only 7 C. In addition to thermal shock and materials compatibility, key feasibility issues for the target are related to its thermal-hydraulic performance. This includes proper flow distribution, flow reversals, possible {open_quotes}hot spots{close_quotes} and the challenge of mitigating the effects of thermal shock through possible injection of helium bubbles throughout the mercury volume or other concepts. The general computational fluid dynamics (CFD) code CFDS-FLOW3D was used to simulate the thermal and flow distribution in three preliminary concepts of the mercury target. Very initial CFD simulation of He bubbles injection demonstrates some potential for simulating behavior of He bubbles in flowing mercury. Much study and development will be required to be able to `predict`, even in a crude way, such a complex phenomena. Future direction in both design and R&D is outlined.« less

  15. Interaction Of CO2 Laser Nanosecond Pulse Train With The Metallic Targets In Optical Breakdown Regime

    NASA Astrophysics Data System (ADS)

    Apollonov, V. V.; Firsov, K. N.; Konov, V. I.; Nikitin, P. I.; Prokhorov, A. M.; Silenok, A. S.; Sorochenko, V. R.

    1986-11-01

    In the present paper the electric field and currents in the air-breakdown plasma, produced by the train of nanosecond pulses of TEA-002 - regenerative amplifier near the un-charged targets are studied. The breakdown thresholds and the efficiency of plasma-target heat transmission are also measured. The results of numerical calculations made for increasing of the pulse train contrast with respect to the background in a regenerative amplifier are advanced.

  16. Stabilizing laser energy density on a target during pulsed laser deposition of thin films

    DOEpatents

    Dowden, Paul C.; Jia, Quanxi

    2016-05-31

    A process for stabilizing laser energy density on a target surface during pulsed laser deposition of thin films controls the focused laser spot on the target. The process involves imaging an image-aperture positioned in the beamline. This eliminates changes in the beam dimensions of the laser. A continuously variable attenuator located in between the output of the laser and the imaged image-aperture adjusts the energy to a desired level by running the laser in a "constant voltage" mode. The process provides reproducibility and controllability for deposition of electronic thin films by pulsed laser deposition.

  17. Plasmonic Nanodiamonds – Targeted Core-shell Type Nanoparticles for Cancer Cell Thermoablation

    PubMed Central

    Rehor, Ivan; Lee, Karin L.; Chen, Kevin; Hajek, Miroslav; Havlik, Jan; Lokajova, Jana; Masat, Milan; Slegerova, Jitka; Shukla, Sourabh; Heidari, Hamed; Bals, Sara

    2015-01-01

    Targeted biocompatible nanostructures with controlled plasmonic and morphological parameters are promising materials for cancer treatment based on selective thermal ablation of cells. Here, core-shell plasmonic nanodiamonds consisting of a silica-encapsulated diamond nanocrystal coated in a gold shell is designed and synthesized. The architecture of particles is analyzed and confirmed in detail using 3-dimensional transmission electron microscope tomography. The particles are biocompatibilized using a PEG polymer terminated with bioorthogonally reactive alkyne groups. Azide-modified transferrin is attached to these particles, and their high colloidal stability and successful targeting to cancer cells overexpressing the transferrin receptor is demonstrated. The particles are nontoxic to the cells and they are readily internalized upon binding to the transferrin receptor. The high plasmonic cross section of the particles in the near-infrared region is utilized to quantitatively ablate the cancer cells with a short, one-minute irradiation by a pulse 750-nm laser. PMID:25336437

  18. Evidence that pulsed electric field treatment enhances the cell wall porosity of yeast cells.

    PubMed

    Ganeva, Valentina; Galutzov, Bojidar; Teissie, Justin

    2014-02-01

    The application of rectangular electric pulses, with 0.1-2 ms duration and field intensity of 2.5-4.5 kV/cm, to yeast suspension mediates liberation of cytoplasmic proteins without cell lysis. The aim of this study was to evaluate the effect of pulsed electric field with similar parameters on cell wall porosity of different yeast species. We found that electrically treated cells become more susceptible to lyticase digestion. In dependence on the strain and the electrical conditions, cell lysis was obtained at 2-8 times lower enzyme concentration in comparison with control untreated cells. The increase of the maximal lysis rate was between two and nine times. Furthermore, when applied at low concentration (1 U/ml), the lyticase enhanced the rate of protein liberation from electropermeabilized cells without provoking cell lysis. Significant differences in the cell surface of control and electrically treated cells were revealed by scanning electron microscopy. Data presented in this study allow us to conclude that electric field pulses provoke not only plasma membrane permeabilization, but also changes in the cell wall structure, leading to increased wall porosity.

  19. The target material influence on the current pulse during high power pulsed magnetron sputtering

    NASA Astrophysics Data System (ADS)

    Moens, Filip; Konstantinidis, Stéphanos; Depla, Diederik

    2017-10-01

    The current-time characteristic during high power pulsed magnetron sputtering is measured under identical conditions for seventeen different target materials. Based on physical processes such as gas rarefaction, ion-induced electron emission, and electron impact ionization, two test parameters were derived that significantly correlate with specific features of the current-time characteristic: i) the peak current is correlated to the momentum transfer between the sputtered material and the argon gas, ii) while the observed current plateau after the peak is connected to the metal ionization rate.

  20. Evidence of femtosecond-laser pulse induced cell membrane nanosurgery

    NASA Astrophysics Data System (ADS)

    Katchinskiy, Nir; Godbout, Roseline; Elezzabi, Abdulhakem Y.

    2017-02-01

    The mechanism of femtosecond laser nanosurgical attachment is investigated in the following article. Using sub-10 femtosecond laser pulses with 800 nm central wavelength were used to attach retinoblastoma cells. During the attachment process the cell membrane phospholipid bilayers hemifuse into one shared phospholipid bilayer, at the location of attachment. Transmission electron microscopy was used in order to verify the above hypothesis. Based on the imaging results, it was concluded that the two cell membrane coalesce to form one single shared membrane. The technique of cell-cell attachment via femtosecond laser pulses could potentially serve as a platform for precise cell membrane manipulation. Manipulation of the cellular membrane is valuable for studying diseases such as cancer; where the expression level of plasma proteins on the cell membrane is altered.

  1. Soft-state biomicrofluidic pulse generator for single cell analysis

    NASA Astrophysics Data System (ADS)

    Sabounchi, Poorya; Ionescu-Zanetti, Cristian; Chen, Roger; Karandikar, Manjiree; Seo, Jeonggi; Lee, Luke P.

    2006-05-01

    We present the design, fabrication, and characterization of a soft-state biomicrofluidic pulse generator for single cell analysis. Hydrodynamic cell trapping via lateral microfluidic junctions allows the trapping of single cells from a bulk suspension. Microfluidic injection sites adjacent to the cell-trapping channels enable the pulsed delivery of nanoliter volumes of biochemical reagent. We demonstrated the application and removal of reagent at a frequency of 10Hz with a rise time of less than 33ms and a reagent consumption rate of 0.2nL/s. It is shown that this system operates as a low-pass filter with a cutoff frequency of 7Hz.

  2. Electrical control of calcium oscillations in mesenchymal stem cells using microsecond pulsed electric fields.

    PubMed

    Hanna, Hanna; Andre, Franck M; Mir, Lluis M

    2017-04-20

    Human mesenchymal stem cells are promising tools for regenerative medicine due to their ability to differentiate into many cellular types such as osteocytes, chondrocytes and adipocytes amongst many other cell types. These cells present spontaneous calcium oscillations implicating calcium channels and pumps of the plasma membrane and the endoplasmic reticulum. These oscillations regulate many basic functions in the cell such as proliferation and differentiation. Therefore, the possibility to mimic or regulate these oscillations might be useful to regulate mesenchymal stem cells biological functions. One or several electric pulses of 100 μs were used to induce Ca 2+ spikes caused by the penetration of Ca 2+ from the extracellular medium, through the transiently electropermeabilized plasma membrane, in human adipose mesenchymal stem cells from several donors. Attached cells were preloaded with Fluo-4 AM and exposed to the electric pulse(s) under the fluorescence microscope. Viability was also checked. According to the pulse(s) electric field amplitude, it is possible to generate a supplementary calcium spike with properties close to those of calcium spontaneous oscillations, or, on the contrary, to inhibit the spontaneous calcium oscillations for a very long time compared to the pulse duration. Through that inhibition of the oscillations, Ca 2+ oscillations of desired amplitude and frequency could then be imposed on the cells using subsequent electric pulses. None of the pulses used here, even those with the highest amplitude, caused a loss of cell viability. An easy way to control Ca 2+ oscillations in mesenchymal stem cells, through their cancellation or the addition of supplementary Ca 2+ spikes, is reported here. Indeed, the direct link between the microsecond electric pulse(s) delivery and the occurrence/cancellation of cytosolic Ca 2+ spikes allowed us to mimic and regulate the Ca 2+ oscillations in these cells. Since microsecond electric pulse delivery

  3. Stoichiometry control of complex oxides by sequential pulsed-laser deposition from binary-oxide targets

    DOE PAGES

    Herklotz, Andreas; Dorr, Kathrin; Ward, Thomas Zac; ...

    2015-04-03

    To have precise atomic layer control over interfaces, we examine the growth of complex oxides through the sequential deposition from binary targets by pulsed laser deposition. In situ reflection high-energy electron diffraction (RHEED) is used to control the growth and achieve films with excellent structural quality. The growth from binary oxide targets is fundamentally different from single target growth modes and shows more similarities to shuttered growth by molecular beam epitaxy. The RHEED intensity oscillations of non-stoichiometric growth are consistent with a model of island growth and accumulation of excess material on the surface that can be utilized to determinemore » the correct stoichiometry for growth. Correct monolayer doses can be determined through an envelope frequency in the RHEED intensity oscillations. In order to demonstrate the ability of this growth technique to create complex heterostructures, the artificial n = 2 and 3 Sr n+1Ti nO 3 n+1 Ruddlesden-Popper phases are grown with good long-range order. Furthermore, this method enables the precise unit-cell level control over the structure of perovskite-type oxides, and thus the growth of complex materials with improved structural quality and electronic functionality.« less

  4. Biophysical Studies of Nanosecond Pulsed Electric Field Induced Cell Membrane Permeabilization

    NASA Astrophysics Data System (ADS)

    Wu, Yu-Hsuan

    Nanosecond megavolts-per-meter pulsed electric field (nsPEF) offers a non-invasive manipulation of intracellular organelles and functions of biological cells. Accordingly, nsPEF is a potential technique for biophysical research and cancer therapy, and is of growing interest. Although, the application of nsPEF has shown electroperturbation on cell plasma membranes and intracellular membranes as well, the mechanisms underlying the electropermeabilization are still not clear. In this thesis, we systematically study nsPEFs (5 and 30 ns) induced membrane permeability change in biological cell in-vitro with different pulse parameters. In Chapter 3, we investigate the nsPEF-induced intracellular membrane permeabilization of mitochondria which play key roles in activating apoptosis in mammalian cells. The results show the evidences of nsPEF-induced membrane permeability increase in mitochondria, and suggest that nsPEF is a potential technology for cancer cell ablation without delivery of drug or gene into cells. In Chapter 2, 4 and 6, we study the properties of nsPEF-induced plasma membrane permeabilization. In the beginning, the change of plasma membrane permeability is studied by uptake of YO-PRO-1 and propidium iodide, fluorescent dyes specifically used as indicators of plasma membrane permeabilization. However, the detection is limited by the fluorescent emission efficiency and detector capability. To increase the detection sensitivity, we later develop a method based on cell volume change due to regulation of osmotic balance that causes water and small ions transport through plasma membrane. We find that even a single 10 MV/m pulse of 5 ns duration produces measureable cell swelling. The results demonstrate that cell swelling is susceptible to nsPEF and can detect membrane permeabilization more easily and precisely than fluorescent dyes. We compare the effects of different pulse parameters (pulse duration, pulse number, electric field amplitude and pulse repetition

  5. Proton acceleration by a pair of successive ultraintense femtosecond laser pulses

    NASA Astrophysics Data System (ADS)

    Ferri, J.; Senje, L.; Dalui, M.; Svensson, K.; Aurand, B.; Hansson, M.; Persson, A.; Lundh, O.; Wahlström, C.-G.; Gremillet, L.; Siminos, E.; DuBois, T. C.; Yi, L.; Martins, J. L.; Fülöp, T.

    2018-04-01

    We investigate the target normal sheath acceleration of protons in thin aluminum targets irradiated at a relativistic intensity by two time-separated ultrashort (35 fs) laser pulses. When the full-energy laser pulse is temporally split into two identical half-energy pulses, and using target thicknesses of 3 and 6 μm, we observe experimentally that the second half-pulse boosts the maximum energy and charge of the proton beam produced by the first half-pulse for time delays below ˜0.6-1 ps. Using two-dimensional particle-in-cell simulations, we examine the variation of the proton energy spectra with respect to the time-delay between the two pulses. We demonstrate that the expansion of the target front surface caused by the first pulse significantly enhances the hot-electron generation by the second pulse arriving after a few hundreds of fs time delay. This enhancement, however, does not suffice to further accelerate the fastest protons driven by the first pulse once three-dimensional quenching effects have set in. This implies a limit to the maximum time delay that leads to proton energy enhancement, which we theoretically determine.

  6. XUV generation from the interaction of pico- and nanosecond laser pulses with nanostructured targets

    NASA Astrophysics Data System (ADS)

    Barte, Ellie Floyd; Lokasani, Ragava; Proska, Jan; Stolcova, Lucie; Maguire, Oisin; Kos, Domagoj; Sheridan, Paul; O'Reilly, Fergal; Sokell, Emma; McCormack, Tom; O'Sullivan, Gerry; Dunne, Padraig; Limpouch, Jiri

    2017-05-01

    Laser-produced plasmas are intense sources of XUV radiation that can be suitable for different applications such as extreme ultraviolet lithography, beyond extreme ultraviolet lithography and water window imaging. In particular, much work has focused on the use of tin plasmas for extreme ultraviolet lithography at 13.5 nm. We have investigated the spectral behavior of the laser produced plasmas formed on closely packed polystyrene microspheres and porous alumina targets covered by a thin tin layer in the spectral region from 2.5 to 16 nm. Nd:YAG lasers delivering pulses of 170 ps (Ekspla SL312P )and 7 ns (Continuum Surelite) duration were focused onto the nanostructured targets coated with tin. The intensity dependence of the recorded spectra was studied; the conversion efficiency (CE) of laser energy into the emission in the 13.5 nm spectral region was estimated. We have observed an increase in CE using high intensity 170 ps Nd:YAG laser pulses as compared with a 7 ns pulse.

  7. Nanosecond pulsed electric field (nsPEF) enhance cytotoxicity of cisplatin to hepatocellular cells by microdomain disruption on plasma membrane.

    PubMed

    Yin, Shengyong; Chen, Xinhua; Xie, Haiyang; Zhou, Lin; Guo, Danjing; Xu, Yuning; Wu, Liming; Zheng, Shusen

    2016-08-15

    Previous studies showed nanosecond pulsed electric field (nsPEF) can ablate solid tumors including hepatocellular carcinoma (HCC) but its effect on cell membrane is not fully understood. We hypothesized nsPEF disrupt the microdomains on outer-cellular membrane with direct mechanical force and as a result the plasma membrane permeability increases to facilitate the small molecule intake. Three HCC cells were pulsed one pulse per minute, an interval longer than nanopore resealing time. The cationized ferritin was used to mark up the electronegative microdomains, propidium iodide (PI) for membrane permeabilization, energy dispersive X-ray spectroscopy (EDS) for the negative cell surface charge and cisplatin for inner-cellular cytotoxicity. We demonstrated that the ferritin marked-microdomain and negative cell surface charge were disrupted by nsPEF caused-mechanical force. The cell uptake of propidium and cytotoxicity of DNA-targeted cisplatin increased with a dose effect. Cisplatin gains its maximum inner-cellular cytotoxicity when combining with nsPEF stimulation. We conclude that nsPEF disrupt the microdomains on the outer cellular membrane directly and increase the membrane permeabilization for PI and cisplatin. The microdomain disruption and membrane infiltration changes are caused by the mechanical force from the changes of negative cell surface charge. Copyright © 2016 Elsevier Inc. All rights reserved.

  8. Effects of electron recirculation on a hard x-ray source observed during the interaction of a high intensity laser pulse with thin Au targets

    NASA Astrophysics Data System (ADS)

    Compant La Fontaine, A.; Courtois, C.; Lefebvre, E.; Bourgade, J. L.; Landoas, O.; Thorp, K.; Stoeckl, C.

    2013-12-01

    The interaction of a high intensity laser pulse on the preplasma of a high-Z solid target produced by the pulse's pedestal generates high-energy electrons. These electrons subsequently penetrate inside the solid target and produce bremsstrahlung photons, generating an x-ray source which can be used for photonuclear studies or to radiograph high area density objects. The source characteristics are compared for targets with thin (20 μm) and thick (100 μm) Au foils on the Omega EP laser at Laboratory for Laser Energetics. Simulations using the particle-in-cell code CALDER show that for a 20 μm thickness Au target, electrons perform multiple round-trips in the target under the effect of the laser ponderomotive potential and the target electrostatic potential. These relativistic electrons have random transverse displacements, with respect to the target normal, attributed to electrostatic fluctuation fields. As a result, the x-ray spot size is increased by a factor 2 for thin target compared to thick targets, in agreement with experimental results. In addition, the computed doses agree with the measured ones provided that electron recirculation in the thin target is taken into account. A dose increase by a factor 1.7 is then computed by allowing for recirculation. In the 100 μm target case, on the other hand, this effect is found to be negligible.

  9. Picosecond pulsed electric fields induce apoptosis in HeLa cells via the endoplasmic reticulum stress and caspase-dependent signaling pathways.

    PubMed

    Chen, Wen-Juan; Xiong, Zheng-Ai; Zhang, Min; Yao, Chen-Guo; Zhao, Zhong-Yong; Hua, Yuan-Yuan; Zhou, Wei

    2013-03-01

    The non-invasive treatment of tumors with preserved fertility holds great promise. The application of pulsed electric field (PEF) is a new biomedical engineering technique for tumor therapy. Picosecond pulsed electric fields (psPEF) can be transferred to target deep tissue non-invasively and precisely; however, research of the biological effects of psPEF on cells is limited. Electric theory predicts that when the pulse duration decreases to nanoseconds and picoseconds, it will mainly affect organelles and lead to intracellular electromanipulations. Previous studies have shown that psPEF targets the mitochondria and induces apoptosis through a mitochondrial-mediated pathway in HeLa cells. The endoplasmic reticulum is also involved in the intrinsic pathways of apoptosis. In the present study, HeLa cells were exposed to psPEF to investigate the underlying mechanisms of apoptosis. MTT assay demonstrated that psPEF displayed strong growth inhibitory effects on HeLa cells. Treatment with psPEF led to marked cell apoptosis and cell cycle arrest at the G2/M phase. In addition, psPEF affected the phosphorylation levels of endoplasmic reticulum sensors and upregulated the expression of glucose-regulated protein 78 (GRP78), glucose-regulated protein 94 (GRP94) and CCAAT enhancer-binding protein (C/EBP) homologous protein (CHOP). These changes were accompanied by the elevation of intracellular Ca2+ concentrations. Furthermore, the activation of caspase-12, -9 and -3, led to the release of cytochrome c, as well as the upregulation of Bax and the downregulation of Bcl-2, as observed in the HeLa cells. Taken together, these data suggest that psPEF is an efficient apoptosis-inducing agent for HeLa cells, which exerts its effects, at least partially, via the endoplasmic reticulum stress and caspase-dependent signaling pathways.

  10. Nanosecond pulsed electric field induced changes in cell surface charge density.

    PubMed

    Dutta, Diganta; Palmer, Xavier-Lewis; Asmar, Anthony; Stacey, Michael; Qian, Shizhi

    2017-09-01

    This study reports that the surface charge density changes in Jurkat cells with the application of single 60 nanosecond pulse electric fields, using atomic force microscopy. Using an atomic force microscope tip and Jurkat cells on silica in a 0.01M KCl ionic concentration, we were able to measure the interfacial forces, while also predicting surface charge densities of both Jurkat cell and silica surfaces. The most important finding is that the pulsing conditions varyingly reduced the cells' surface charge density. This offers a novel way in which to examine cellular effects of pulsed electric fields that may lead to the identification of unique mechanical responses. Compared to a single low field strength NsPEF (15kV/cm) application, exposure of Jurkat cells to a single high field strength NsPEF (60kV/cm) resulted in a further reduction in charge density and major morphological changes. The structural, physical, and chemical properties of biological cells immensely influence their electrostatic force; we were able to investigate this through the use of atomic force microscopy by measuring the surface forces between the AFM's tip and the Jurkat cells under different pulsing conditions as well as the interfacial forces in ionic concentrations. Copyright © 2017 Elsevier Ltd. All rights reserved.

  11. A pulsed supersonic gas jet target for precision spectroscopy at the HITRAP facility at GSI

    NASA Astrophysics Data System (ADS)

    Tiedemann, D.; Stiebing, K. E.; Winters, D. F. A.; Quint, W.; Varentsov, V.; Warczak, A.; Malarz, A.; Stöhlker, Th.

    2014-11-01

    A pulsed supersonic gas jet target for experiments at the HITRAP facility at the GSI Helmholtzzentrum für Schwerionenforschung in Darmstadt has been designed and built as a multi-purpose installation for key experiments on fundamental atomic physics in strong fields. This setup is currently installed at the Institut für Kernphysik of Goethe-University, Frankfurt am Main (IKF), in order to explore its operation prior to its installation at the HITRAP facility. Design and performance of the target are described. The measured target densities of 5.9×1012 atoms/cm3 for helium and 8.1×1012 atoms/cm³ for argon at the stagnation pressure of 30 bar match the required values. The target-beam diameter of 0.9 mm and the pulsed operation mode (jet built-up-time ≤15 ms) are well suited for the use at HITRAP.

  12. Dynamic target ionization using an ultrashort pulse of a laser field

    NASA Astrophysics Data System (ADS)

    Makarov, D. N.; Matveev, V. I.; Makarova, K. A.

    2014-09-01

    Ionization processes under the interaction of an ultrashort pulse of an electromagnetic field with atoms in nonstationary states are considered. As an example, the ionization probability of the hydrogen-like atom upon the decay of quasi-stationary state is calculated. The method developed can be applied to complex systems, including targets in collisional states and various chemical reactions.

  13. Effects of Pulsed Electromagnetic Fields on Breast Cancer Cell Line MCF 7 Using Absorption Spectroscopy.

    PubMed

    Alcantara, Dominic Z; Soliman, Ian Jerry S; Pobre, Romeric F; Naguib, Raouf N G

    2017-07-01

    We present an analysis of the effects of pulsed electromagnetic fields (PEMF) with 3.3 MHz carrier frequency and modulated by audio resonant frequencies on the MCF-7 breast cancer cell line in vitro using absorption spectroscopy. This involves a fluorescence dye called PrestoBlue™ Cell Viability Reagent and a spectrophotometry to test the viability of MCF-7 breast cancer cells under different PEMF treatment conditions in terms of the cell absorption values. The DNA molecule of the MCF-7 breast cancer cells has an electric dipole property that renders it sensitive and reactive to applied electromagnetic fields. Resonant frequencies derived from four genes mutated in MCF-7 breast cancer cells [rapamycin-insensitive companion of mammalian target of rapamycin (RICTOR), peroxisome proliferator-activated receptor (PPARG), Nijmegen breakage syndrome 1 (NBN) and checkpoint kinase 2 (CHEK2)] were applied in generating square pulsed electromagnetic waves. Effects were monitored through measurement of absorption of the samples with PrestoBlue™, and the significance of the treatment was determined using the t-test. There was a significant effect on MCF-7 cells after treatment with PEMF at the resonant frequencies of the following genes for specific durations of exposure: RICTOR for 10 min, PPARG for 10 min, NBN for 15 min, and CHEK2 for 5 min. Copyright© 2017, International Institute of Anticancer Research (Dr. George J. Delinasios), All rights reserved.

  14. Thin-film preparation by back-surface irradiation pulsed laser deposition using metal powder targets

    NASA Astrophysics Data System (ADS)

    Kawasaki, Hiroharu; Ohshima, Tamiko; Yagyu, Yoshihito; Ihara, Takeshi; Yamauchi, Makiko; Suda, Yoshiaki

    2017-01-01

    Several kinds of functional thin films were deposited using a new thin-film preparation method named the back-surface irradiation pulsed laser deposition (BIPLD) method. In this BIPLD method, powder targets were used as the film source placed on a transparent target holder, and then a visible-wavelength pulsed laser was irradiated from the holder side to the substrate. Using this new method, titanium oxide and boron nitride thin films were deposited on the silicon substrate. Surface scanning electron microscopy (SEM) images suggest that all of the thin films were deposited on the substrate with some large droplets irrespective of the kind of target used. The deposition rate of the films prepared by using this method was calculated from film thickness and deposition time to be much lower than that of the films prepared by conventional PLD. X-ray diffraction (XRD) measurement results suggest that rutile and anatase TiO2 crystal peaks were formed for the films prepared using the TiO2 rutile powder target. Crystal peaks of hexagonal boron nitride were observed for the films prepared using the boron nitride powder target. The crystallinity of the prepared films was changed by annealing after deposition.

  15. Targeting dendritic cells--why bother?

    PubMed

    Kreutz, Martin; Tacken, Paul J; Figdor, Carl G

    2013-04-11

    Vaccination is among the most efficient forms of immunotherapy. Although sometimes inducing lifelong protective B-cell responses, T-cell-mediated immunity remains challenging. Targeting antigen to dendritic cells (DCs) is an extensively explored concept aimed at improving cellular immunity. The identification of various DC subsets with distinct functional characteristics now allows for the fine-tuning of targeting strategies. Although some of these DC subsets are regarded as superior for (cross-) priming of naive T cells, controversies still remain about which subset represents the best target for immunotherapy. Because targeting the antigen alone may not be sufficient to obtain effective T-cell responses, delivery systems have been developed to target multiple vaccine components to DCs. In this Perspective, we discuss the pros and cons of targeting DCs: if targeting is beneficial at all and which vaccine vehicles and immunization routes represent promising strategies to reach and activate DCs.

  16. EWS/FLI-l peptide-pulsed dendritic cells induces the antitumor immunity in a murine Ewing's sarcoma cell model.

    PubMed

    Peng, Wei; Huang, Xunwu; Yang, Dazhi

    2014-08-01

    An increasing number of T-cell epitopes derived from various tumor-associated antigens have been reported, and they proved to play significant roles for tumor rejection both in vivo and in vitro. Over 85% of Ewing's sarcoma family of tumors (ESFTs) express tumor-specific chimeric protein EWS/FLI-1, making it an attractive target for therapeutic cytotoxic T-lymphocyte responses. Here, we identified a novel peptide epitope derived from the EWS/FLI-1 protein and demonstrated that effectors induced by the peptide could specifically secrete IFN-γ and lyse the tumor cell line of EWS/FLI-1-positive and HLA-matched cells. In addition, mice treated with dendritic cells pulsed with the EWS/FLI-1 epitope were able to reject a lethal tumor inoculation of the Ewing's sarcoma A673 cells. Therefore, these data provide evidence for the use of the EWS/FLI-l peptide epitope in T cell-based immunotherapeutic concepts against Ewing's sarcoma cell in vitro and in vivo. Copyright © 2014 Elsevier B.V. All rights reserved.

  17. Characterization of femtosecond-laser pulse induced cell membrane nanosurgical attachment.

    PubMed

    Katchinskiy, Nir; Godbout, Roseline; Elezzabi, Abdulhakem Y

    2016-07-01

    This article provides insight into the mechanism of femtosecond laser nanosurgical attachment of cells. We have demonstrated that during the attachment of two retinoblastoma cells using sub-10 femtosecond laser pulses, with 800 nm central wavelength, the phospholipid molecules of both cells hemifuse and form one shared phospholipid bilayer, at the attachment location. In order to verify the hypothesis that hemifusion takes place, transmission electron microscope images of the cell membranes of retinoblastoma cells were taken. It is shown that at the attachment interface, the two cell membranes coalesce and form one single membrane shared by both cells. Thus, further evidence is provided to support the hypothesis that laser-induced ionization process led to an ultrafast reversible destabilization of the phospholipid layer of the cellular membrane, which resulted in cross-linking of the phospholipid molecules in each membrane. This process of hemifusion occurs throughout the entire penetration depth of the femtosecond laser pulse train. Thus, the attachment between the cells takes place across a large surface area, which affirms our findings of strong physical attachment between the cells. The femtosecond laser pulse hemifusion technique can potentially provide a platform for precise molecular manipulation of cellular membranes. Manipulation of the cellular membrane is an important procedure that could aid in studying diseases such as cancer; where the expression level of plasma proteins on the cell membrane is altered.

  18. Pulse labeling of RNA of mammalian cells.

    PubMed

    Rovera, G; Berman, S; Baserga, R

    1970-04-01

    When cells from a hypotetraploid strain of Ehrlich ascites tumor are exposed to uridine-(3)H either in vivo or in vitro, the amount of radioactivity incorporated into RNA reaches a maximum within ten minutes, after which any further incorporation stops. (3)H-uridine triphosphate disappears from the acid soluble pool within 30 minutes and the findings indicate that the RNA of these cells can be pulse labeled without the use of any antibiotic or the need of a "chase." The stability of the pulse labeled RNA in the presence of pentobarbital (an inhibitor of RNA synthesis) indicates the virtual absence of RNA breakdown. However, actinomycin D, at a dosage of 250 mug/mouse in vivo and 10 mug/ml in vitro produces breakdown of labeled RNA, thus confirming earlier observations that the drug is not a suitable tool for RNA kinetics determinations. The pulse-labeled RNA leaves the nucleus slowly and some radioactive RNA is still present in the nuclear fraction after 24 hours. Radioactivity begins to appear in cytoplasmic ribosomal RNA after 20 minutes and continues to increase up to six hours.

  19. Macrophage and tumor cell responses to repetitive pulsed X-ray radiation

    NASA Astrophysics Data System (ADS)

    Buldakov, M. A.; Tretyakova, M. S.; Ryabov, V. B.; Klimov, I. A.; Kutenkov, O. P.; Kzhyshkowska, J.; Bol'shakov, M. A.; Rostov, V. V.; Cherdyntseva, N. V.

    2017-05-01

    To study a response of tumor cells and macrophages to the repetitive pulsed low-dose X-ray radiation. Methods. Tumor growth and lung metastasis of mice with an injected Lewis lung carcinoma were analysed, using C57Bl6. Monocytes were isolated from a human blood, using CD14+ magnetic beads. IL6, IL1-betta, and TNF-alpha were determined by ELISA. For macrophage phenotyping, a confocal microscopy was applied. “Sinus-150” was used for the generation of pulsed X-ray radiation (the absorbed dose was below 0.1 Gy, the pulse repetition frequency was 10 pulse/sec). The irradiation of mice by 0.1 Gy pulsed X-rays significantly inhibited the growth of primary tumor and reduced the number of metastatic colonies in the lung. Furthermore, the changes in macrophage phenotype and cytokine secretion were observed after repetitive pulsed X-ray radiation. Conclusion. Macrophages and tumor cells had a different response to a low-dose pulsed X-ray radiation. An activation of the immune system through changes of a macrophage phenotype can result in a significant antitumor effect of the low-dose repetitive pulsed X-ray radiation.

  20. Stoichiometry control of complex oxides by sequential pulsed-laser deposition from binary-oxide targets

    DOE PAGES

    Herklotz, A.; Dörr, Kathrin; Ward, T. Z.; ...

    2015-04-03

    In this paper, to have precise atomic layer control over interfaces, we examine the growth of complex oxides through the sequential deposition from binary targets by pulsed laser deposition. In situ reflection high-energy electron diffraction (RHEED) is used to control the growth and achieve films with excellent structural quality. The growth from binary oxide targets is fundamentally different from single target growth modes and shows more similarities to shuttered growth by molecular beam epitaxy. The RHEED intensity oscillations of non-stoichiometric growth are consistent with a model of island growth and accumulation of excess material on the surface that can bemore » utilized to determine the correct stoichiometry for growth. Correct monolayer doses can be determined through an envelope frequency in the RHEED intensity oscillations. In order to demonstrate the ability of this growth technique to create complex heterostructures, the artificial n = 2 and 3 Sr n +1Ti n O 3 n +1 Ruddlesden-Popper phases are grown with good long-range order. Finally, this method enables the precise unit-cell level control over the structure of perovskite-type oxides, and thus the growth of complex materials with improved structural quality and electronic functionality.« less

  1. Avoiding the side effects of electric current pulse application to electroporated cells in disposable small volume cuvettes assures good cell survival.

    PubMed

    Grys, Maciej; Madeja, Zbigniew; Korohoda, Włodzimierz

    2017-01-01

    The harmful side effects of electroporation to cells due to local changes in pH, the appearance of toxic electrode products, temperature increase, and the heterogeneity of the electric field acting on cells in the cuvettes used for electroporation were observed and discussed in several laboratories. If cells are subjected to weak electric fields for prolonged periods, for example in experiments on cell electrophoresis or galvanotaxis the same effects are seen. In these experiments investigators managed to reduce or eliminate the harmful side effects of electric current application. For the experiments, disposable 20 μl cuvettes with two walls made of dialysis membranes were constructed and placed in a locally focused electric field at a considerable distance from the electrodes. Cuvettes were mounted into an apparatus for horizontal electrophoresis and the cells were subjected to direct current electric field (dcEF) pulses from a commercial pulse generator of exponentially declining pulses and from a custom-made generator of double and single rectangular pulses. More than 80% of the electroporated cells survived the dcEF pulses in both systems. Side effects related to electrodes were eliminated in both the flow through the dcEF and in the disposable cuvettes placed in the focused dcEFs. With a disposable cuvette system, we also confirmed the sensitization of cells to a dcEF using procaine by observing the loading of AT2 cells with calceine and using a square pulse generator, applying 50 ms single rectangular pulses. We suggest that the same methods of avoiding the side effects of electric current pulse application as in cell electrophoresis and galvanotaxis should also be used for electroporation. This conclusion was confirmed in our electroporation experiments performed in conditions assuring survival of over 80% of the electroporated cells. If the amplitude, duration, and shape of the dcEF pulse are known, then electroporation does not depend on the type of

  2. A table-top monochromator for tunable femtosecond XUV pulses generated in a semi-infinite gas cell: Experiment and simulations

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    Conta, A. von; Huppert, M.; Wörner, H. J.

    2016-07-15

    We present a new design of a time-preserving extreme-ultraviolet (XUV) monochromator using a semi-infinite gas cell as a source. The performance of this beamline in the photon-energy range of 20 eV–42 eV has been characterized. We have measured the order-dependent XUV pulse durations as well as the flux and the spectral contrast. XUV pulse durations of ≤40 fs using 32 fs, 800 nm driving pulses were measured on the target. The spectral contrast was better than 100 over the entire energy range. A simple model based on the strong-field approximation is presented to estimate different contributions to the measured XUVmore » pulse duration. On-axis phase-matching calculations are used to rationalize the variation of the photon flux with pressure and intensity.« less

  3. Natural Killer Cell Immunotherapy Targeting Cancer Stem Cells

    PubMed Central

    Luna, Jesus I; Grossenbacher, Steven K.; Murphy, William J; Canter, Robert J

    2017-01-01

    Introduction Standard cytoreductive cancer therapy, such as chemotherapy and radiotherapy, are frequently resisted by a small portion of cancer cells with “stem-cell” like properties including quiescence and repopulation. Immunotherapy represents a breakthrough modality for improving oncologic outcomes in cancer patients. Since the success of immunotherapy is not contingent on target cell proliferation, it may also be uniquely suited to address the problem of resistance and repopulation exerted by cancer stem cells (CSCs). Areas covered Natural killer (NK) cells have long been known for their ability to reject allogeneic hematopoietic stem cells, and there are increasing data demonstrating that NK cells can selectively identify and lyse CSCs. In this report, we review the current knowledge of CSCs and NK cells and highlight recent studies that support the concept that NK cells are capable of targeting CSC in solid tumors, especially in the context of combination therapy simultaneously targeting non-CSCs and CSCs. Expert Opinion Unlike cytotoxic cancer treatments, NK cells are able to target and eliminate quiescent/non-proliferating cells such as CSCs, and these enigmatic cells are an important source of relapse and metastasis. NK targeting of CSCs represents a novel and potentially high impact method to capitalize on the intrinsic therapeutic potential of NK cells. PMID:27960589

  4. Monoenergetic acceleration of a target foil by circularly polarized laser pulse in RPA regime without thermal heating

    NASA Astrophysics Data System (ADS)

    Khudik, V.; Yi, S. A.; Siemon, C.; Shvets, G.

    2012-12-01

    A kinetic model of the monoenergetic acceleration of a target foil irradiated by the circularly polarized laser pulse is developed. The target moves without thermal heating with constant acceleration which is provided by chirping the frequency of the laser pulse and correspondingly increasing its intensity. In the accelerated reference frame, bulk plasma in the target is neutral and its parameters are stationary: cold ions are immobile while nonrelativistic electrons bounce back and forth inside the potential well formed by ponderomotive and electrostatic potentials. It is shown that a positive charge left behind of the moving target in the ion tail and a negative charge in front of the target in the electron sheath form a capacitor whose constant electric field accelerates the ions of the target. The charge separation is maintained by the radiation pressure pushing electrons forward. The scalings of the target thickness and electromagnetic radiation with the electron temperature are found.

  5. Dendritic Cells Pulsed with Leukemia Cell-Derived Exosomes More Efficiently Induce Antileukemic Immunities

    PubMed Central

    Wei, Wei; Shen, Chang; Deng, Xiaohui; Chen, Linjun; Ma, Liyuan; Hao, Siguo

    2014-01-01

    Dendritic cells (DCs) and tumor cell-derived exosomes have been used to develop antitumor vaccines. However, the biological properties and antileukemic effects of leukemia cell-derived exosomes (LEXs) are not well described. In this study, the biological properties and induction of antileukemic immunity of LEXs were investigated using transmission electron microscopy, western blot analysis, cytotoxicity assays, and animal studies. Similar to other tumor cells, leukemia cells release exosomes. Exosomes derived from K562 leukemia cells (LEXK562) are membrane-bound vesicles with diameters of approximately 50–100 μm and harbor adhesion molecules (e.g., intercellular adhesion molecule-1) and immunologically associated molecules (e.g., heat shock protein 70). In cytotoxicity assays and animal studies, LEXs-pulsed DCs induced an antileukemic cytotoxic T-lymphocyte immune response and antileukemic immunity more effectively than did LEXs and non-pulsed DCs (P<0.05). Therefore, LEXs may harbor antigens and immunological molecules associated with leukemia cells. As such, LEX-based vaccines may be a promising strategy for prolonging disease-free survival in patients with leukemia after chemotherapy or hematopoietic stem cell transplantation. PMID:24622345

  6. Pulsed laser triggered high speed microfluidic fluorescence activated cell sorter†‡

    PubMed Central

    Wu, Ting-Hsiang; Chen, Yue; Park, Sung-Yong; Hong, Jason; Teslaa, Tara; Zhong, Jiang F.; Di Carlo, Dino; Teitell, Michael A.

    2014-01-01

    We report a high speed and high purity pulsed laser triggered fluorescence activated cell sorter (PLACS) with a sorting throughput up to 20 000 mammalian cells s−1 with 37% sorting purity, 90% cell viability in enrichment mode, and >90% purity in high purity mode at 1500 cells s−1 or 3000 beads s−1. Fast switching (30 μs) and a small perturbation volume (~90 pL) is achieved by a unique sorting mechanism in which explosive vapor bubbles are generated using focused laser pulses in a single layer microfluidic PDMS channel. PMID:22361780

  7. Modeling of high efficiency solar cells under laser pulse for power beaming applications

    NASA Technical Reports Server (NTRS)

    Jain, Raj K.; Landis, Geoffrey A.

    1994-01-01

    Solar cells have been used to convert sunlight to electrical energy for many years and also offer great potential for non-solar energy conversion applications. Their greatly improved performance under monochromatic light compared to sunlight, makes them suitable as photovoltaic (PV) receivers in laser power beaming applications. Laser beamed power to a PV array receiver could provide power to satellites, an orbital transfer vehicle, or a lunar base. Gallium arsenide (GaAs) and indium phosphide (InP) solar cells have calculated efficiencies of more than 50 percent under continuous illumination at the optimum wavelength. Currently high power free-electron lasers are being developed which operate in pulsed conditions. Understanding cell behavior under a laser pulse is important in the selection of the solar cell material and the laser. An experiment by NAsA lewis and JPL at the AVLIS laser facility in Livermore, CA presented experimental data on cell performance under pulsed laser illumination. Reference 5 contains an overview of technical issues concerning the use of solar cells for laser power conversion, written before the experiments were performed. As the experimental results showed, the actual effects of pulsed operation are more complicated. Reference 6 discusses simulations of the output of GaAs concentrator solar cells under pulsed laser illumination. The present paper continues this work, and compares the output of Si and GaAs solar cells.

  8. Hundred joules plasma focus device as a potential pulsed source for in vitro cancer cell irradiation

    NASA Astrophysics Data System (ADS)

    Jain, J.; Moreno, J.; Andaur, R.; Armisen, R.; Morales, D.; Marcelain, K.; Avaria, G.; Bora, B.; Davis, S.; Pavez, C.; Soto, L.

    2017-08-01

    Plasma focus devices may arise as useful source to perform experiments aimed to study the effects of pulsed radiation on human cells in vitro. In the present work, a table top hundred joules plasma focus device, namely "PF-400J", was adapted to irradiate colorectal cancer cell line, DLD-1. For pulsed x-rays, the doses (energy absorbed per unit mass, measured in Gy) were measured using thermoluminescence detectors (TLD-100 dosimeters). The neutron fluence and the average energy were used to estimate the pulsed neutron doses. Fifty pulses of x-rays (0.12 Gy) and fifty pulses of neutrons (3.5 μGy) were used to irradiate the cancer cells. Irradiation-induced DNA damage and cell death were assessed at different time points after irradiation. Cell death was observed using pulsed neutron irradiation, at ultralow doses. Our results indicate that the PF-400J can be used for in vitro assessment of the effect of pulsed radiation in cancer cell research.

  9. [The electroporation effects of high power pulse microwave and electromagnetic pulse irradiation on the membranes of cardiomyocyte cells and the mechanism therein involved].

    PubMed

    Deng, Hua; Wang, Dewen; Peng, Ruiyun; Wang, Shuiming; Chen, Jiankui; Zhang, Sa; Dong, Bo; Wang, Xiaomin

    2005-08-01

    Though there is ongoing public concern on potential hazards and risk of electromagnetic radiation, the bioeffects mechanism of electromagnetic fields remains obscure. Heart is one of the organs susceptive to electromagnetic fields (EMF). This study was designed to assess the influence of high power pulse microwave and electromagnetic pulse irradiation on cardiomyocytes, to explore the critical mechanism of electromagnetic fields, and to explain the regular course of injury caused by exposure to pulse EMF. Cultured cardiomyocytes were irradiated by high power pulse microwave and electromagnetic pulse first, then a series of apparatus including atom force microscope, laser scanning confocal microscope and flow cytometer were used to examine the changes of cell membrane conformation, structure and function. After irradiation, the cardiomyocytes pulsated slower or stop, the cells conformation was abnormal, the cells viability declined, and the percentage of apoptosis and necrosis increased significantly (P< 0.01). The cell membrane had pores unequal in size, and lost its penetration character. The concentration of Na+, K+, Ca2+, Cl-, Mg2+, Ca2+ and P3+ in cell culture medium increased significantly (P< 0.01). and the concentration of Ca2+ in cells ([Ca2+]i) decreased significantly (P<0.01). The results indicated that cardiomyocytes are susceptible to non-ionizing radiation. Pulse electromagnetic field can induce cardiomyocytes electroporation, and can do great damage to cells conformation, structure and function. Electroporation is one of the most critical mechanisms to explain the athermal effects of electromagnetic radiation.

  10. Multi probes measurements at the PALS Facility Research Centre during high intense laser pulse interactions with various target materials

    NASA Astrophysics Data System (ADS)

    De Marco, Massimo; Krása, Josef; Cikhardt, Jakub; Consoli, Fabrizio; De Angelis, Riccardo; Pfeifer, Miroslav; Krůs, Miroslav; Dostál, Jan; Margarone, Daniele; Picciotto, Antonino; Velyhan, Andriy; Klír, Daniel; Dudžák, Roman; Limpouch, Jiří; Korn, Georg

    2018-01-01

    During the interaction of high intense laser pulse with solid target, a large amount of hot electrons is produced and a giant Electromagnetic Pulse (EMP) is generated due to the current flowing into the system target-target holder, as well as due to the escaping charged particles in vacuum. EMP production for different target materials is investigated inside and outside the target chamber, using monopole antenna, super wide-band microstrip antenna and Moebius antenna. The EMP consists in a fast transient magnetic field lasting hundreds of nanosecond with frequencies ranging from MHz to tens of GHz. Measurements of magnetic field and return target current in the range of kA were carried out by an inductive target probe (Cikhardt J. et al. Rev. Sci. Instrum. 85 (2014) 103507).

  11. Membrane permeabilization of mammalian cells using bursts of high magnetic field pulses

    PubMed Central

    Grainys, Audrius; Kranjc, Matej; Miklavčič, Damijan

    2017-01-01

    Background Cell membrane permeabilization by pulsed electromagnetic fields (PEMF) is a novel contactless method which results in effects similar to conventional electroporation. The non-invasiveness of the methodology, independence from the biological object homogeneity and electrical conductance introduce high flexibility and potential applicability of the PEMF in biomedicine, food processing, and biotechnology. The inferior effectiveness of the PEMF permeabilization compared to standard electroporation and the lack of clear description of the induced transmembrane transport are currently of major concern. Methods The PEMF permeabilization experiments have been performed using a 5.5 T, 1.2 J pulse generator with a multilayer inductor as an applicator. We investigated the feasibility to increase membrane permeability of Chinese Hamster Ovary (CHO) cells using short microsecond (15 µs) pulse bursts (100 or 200 pulses) at low frequency (1 Hz) and high dB/dt (>106 T/s). The effectiveness of the treatment was evaluated by fluorescence microscopy and flow cytometry using two different fluorescent dyes: propidium iodide (PI) and YO-PRO®-1 (YP). The results were compared to conventional electroporation (single pulse, 1.2 kV/cm, 100 µs), i.e., positive control. Results The proposed PEMF protocols (both for 100 and 200 pulses) resulted in increased number of permeable cells (70 ± 11% for PI and 67 ± 9% for YP). Both cell permeabilization assays also showed a significant (8 ± 2% for PI and 35 ± 14% for YP) increase in fluorescence intensity indicating membrane permeabilization. The survival was not affected. Discussion The obtained results demonstrate the potential of PEMF as a contactless treatment for achieving reversible permeabilization of biological cells. Similar to electroporation, the PEMF permeabilization efficacy is influenced by pulse parameters in a dose-dependent manner. PMID:28462057

  12. Oxygen targeting in preterm infants using the Masimo SET Radical pulse oximeter.

    PubMed

    Johnston, Ewen D; Boyle, Breidge; Juszczak, Ed; King, Andy; Brocklehurst, Peter; Stenson, Ben J

    2011-11-01

    A pretrial clinical improvement project for the BOOST-II UK trial of oxygen saturation targeting revealed an artefact affecting saturation profiles obtained from the Masimo Set Radical pulse oximeter. Saturation was recorded every 10 s for up to 2 weeks in 176 oxygen dependent preterm infants in 35 UK and Irish neonatal units between August 2006 and April 2009 using Masimo SET Radical pulse oximeters. Frequency distributions of % time at each saturation were plotted. An artefact affecting the saturation distribution was found to be attributable to the oximeter's internal calibration algorithm. Revised software was installed and saturation distributions obtained were compared with four other current oximeters in paired studies. There was a reduction in saturation values of 87-90%. Values above 87% were elevated by up to 2%, giving a relative excess of higher values. The software revision eliminated this, improving the distribution of saturation values. In paired comparisons with four current commercially available oximeters, Masimo oximeters with the revised software returned similar saturation distributions. A characteristic of the software algorithm reduces the frequency of saturations of 87-90% and increases the frequency of higher values returned by the Masimo SET Radical pulse oximeter. This effect, which remains within the recommended standards for accuracy, is removed by installing revised software (board firmware V4.8 or higher). Because this observation is likely to influence oxygen targeting, it should be considered in the analysis of the oxygen trial results to maximise their generalisability.

  13. Cooperative tumour cell membrane targeted phototherapy

    NASA Astrophysics Data System (ADS)

    Kim, Heegon; Lee, Junsung; Oh, Chanhee; Park, Ji-Ho

    2017-06-01

    The targeted delivery of therapeutics using antibodies or nanomaterials has improved the precision and safety of cancer therapy. However, the paucity and heterogeneity of identified molecular targets within tumours have resulted in poor and uneven distribution of targeted agents, thus compromising treatment outcomes. Here, we construct a cooperative targeting system in which synthetic and biological nanocomponents participate together in the tumour cell membrane-selective localization of synthetic receptor-lipid conjugates (SR-lipids) to amplify the subsequent targeting of therapeutics. The SR-lipids are first delivered selectively to tumour cell membranes in the perivascular region using fusogenic liposomes. By hitchhiking with extracellular vesicles secreted by the cells, the SR-lipids are transferred to neighbouring cells and further spread throughout the tumour tissues where the molecular targets are limited. We show that this tumour cell membrane-targeted delivery of SR-lipids leads to uniform distribution and enhanced phototherapeutic efficacy of the targeted photosensitizer.

  14. Pulsed Magnetic Field Improves the Transport of Iron Oxide Nanoparticles through Cell Barriers

    PubMed Central

    Min, Kyoung Ah; Shin, Meong Cheol; Yu, Faquan; Yang, Meizhu; David, Allan E.; Yang, Victor C.; Rosania, Gus R.

    2013-01-01

    Understanding how a magnetic field affects the interaction of magnetic nanoparticles (MNPs) with cells is fundamental to any potential downstream applications of MNPs as gene and drug delivery vehicles. Here, we present a quantitative analysis of how a pulsed magnetic field influences the manner in which MNPs interact with, and penetrate across a cell monolayer. Relative to a constant magnetic field, the rate of MNP uptake and transport across cell monolayers was enhanced by a pulsed magnetic field. MNP transport across cells was significantly inhibited at low temperature under both constant and pulsed magnetic field conditions, consistent with an active mechanism (i.e. endocytosis) mediating MNP transport. Microscopic observations and biochemical analysis indicated that, in a constant magnetic field, transport of MNPs across the cells was inhibited due to the formation of large (>2 μm) magnetically-induced MNP aggregates, which exceeded the size of endocytic vesicles. Thus, a pulsed magnetic field enhances the cellular uptake and transport of MNPs across cell barriers relative to a constant magnetic field by promoting accumulation while minimizing magnetically-induced MNP aggregates at the cell surface. PMID:23373613

  15. Effect of pulsed current charging on the performance of nickel-cadium cells

    NASA Technical Reports Server (NTRS)

    Bedrossian, A. A.; Cheh, H. Y.

    1977-01-01

    The effect of pulsed current charging on the charge acceptance of NiCd cells in terms of mass transfer, kinetic, and structural considerations was investigated. A systemic investigation on the performance of Ni-Cd cells by pulsed current charging was conducted under a variety of well-defined charge-discharge conditions. Experiments were carried out with half cells and film electrodes. The system behavior was studied by charge acceptance, mechanistic, and structural measurements.

  16. Conceptual moderator studies for the Spallation Neutron Source short-pulse second target station

    DOE PAGES

    Gallmeier, F. X.; Lu, W.; Riemer, B. W.; ...

    2016-06-14

    We identified candidate moderator configurations for a short-pulse second target station (STS) at the Oak Ridge National Laboratory Spallation Neutron Source (SNS) using a global optimizer framework built around the MCNPX particle transport code. Neutron brightness metrics were selected as the figure-of-merit. We assumed that STS would use one out of six proton pulses produced by an SNS accelerator upgraded to operate at 1.3 GeV proton energy, 2.8 MW power and 60 Hz repetition rate. The simulations indicate that the peak brightness can be increased by a factor of 5 and 2.5 on a per proton pulse basis compared tomore » the SNS first target station for both coupled and decoupled para-hydrogen moderators, respectively. Additional increases by factors of 3 and 2 were demonstrated for coupled and decoupled moderators, respectively, by reducing the area of neutron emission from 100 × 100 mm 2 to 20 × 20 mm 2. Furthermore, this increase in brightness has the potential to translate to an increase of beam intensity at the instruments’ sample positions even though the total neutron emission of the smaller moderator is less than that of the larger. This is especially true for instruments with small samples (beam dimensions). The increased fluxes in the STS moderators come at accelerated poison and de-coupler burnout and higher radiation-induced material damage rates per unit power, which overall translate into lower moderator lifetimes. Our first effort decoupled group moderators into a cluster collectively positioning them at the peak neutron production zone in the target and having a three-port neutron emission scheme that complements that of a cylindrical coupled moderator.« less

  17. Conceptual moderator studies for the Spallation Neutron Source short-pulse second target station

    NASA Astrophysics Data System (ADS)

    Gallmeier, F. X.; Lu, W.; Riemer, B. W.; Zhao, J. K.; Herwig, K. W.; Robertson, J. L.

    2016-06-01

    Candidate moderator configurations for a short-pulse second target station (STS) at the Oak Ridge National Laboratory Spallation Neutron Source (SNS) have been identified using a global optimizer framework built around the MCNPX particle transport code. Neutron brightness metrics were selected as the figure-of-merit. We assumed that STS would use one out of six proton pulses produced by an SNS accelerator upgraded to operate at 1.3 GeV proton energy, 2.8 MW power and 60 Hz repetition rate. The simulations indicate that the peak brightness can be increased by a factor of 5 and 2.5 on a per proton pulse basis compared to the SNS first target station for both coupled and decoupled para-hydrogen moderators, respectively. Additional increases by factors of 3 and 2 were demonstrated for coupled and decoupled moderators, respectively, by reducing the area of neutron emission from 100 × 100 mm2 to 20 × 20 mm2. This increase in brightness has the potential to translate to an increase of beam intensity at the instruments' sample positions even though the total neutron emission of the smaller moderator is less than that of the larger. This is especially true for instruments with small samples (beam dimensions). The increased fluxes in the STS moderators come at accelerated poison and de-coupler burnout and higher radiation-induced material damage rates per unit power, which overall translate into lower moderator lifetimes. A first effort was undertaken to group decoupled moderators into a cluster collectively positioning them at the peak neutron production zone in the target and having a three-port neutron emission scheme that complements that of a cylindrical coupled moderator.

  18. Conceptual moderator studies for the Spallation Neutron Source short-pulse second target station

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    Gallmeier, F. X.; Lu, W.; Riemer, B. W.

    We identified candidate moderator configurations for a short-pulse second target station (STS) at the Oak Ridge National Laboratory Spallation Neutron Source (SNS) using a global optimizer framework built around the MCNPX particle transport code. Neutron brightness metrics were selected as the figure-of-merit. We assumed that STS would use one out of six proton pulses produced by an SNS accelerator upgraded to operate at 1.3 GeV proton energy, 2.8 MW power and 60 Hz repetition rate. The simulations indicate that the peak brightness can be increased by a factor of 5 and 2.5 on a per proton pulse basis compared tomore » the SNS first target station for both coupled and decoupled para-hydrogen moderators, respectively. Additional increases by factors of 3 and 2 were demonstrated for coupled and decoupled moderators, respectively, by reducing the area of neutron emission from 100 × 100 mm 2 to 20 × 20 mm 2. Furthermore, this increase in brightness has the potential to translate to an increase of beam intensity at the instruments’ sample positions even though the total neutron emission of the smaller moderator is less than that of the larger. This is especially true for instruments with small samples (beam dimensions). The increased fluxes in the STS moderators come at accelerated poison and de-coupler burnout and higher radiation-induced material damage rates per unit power, which overall translate into lower moderator lifetimes. Our first effort decoupled group moderators into a cluster collectively positioning them at the peak neutron production zone in the target and having a three-port neutron emission scheme that complements that of a cylindrical coupled moderator.« less

  19. Nike Experiment to Observe Strong Areal Mass Oscillations in a Rippled Target Hit by a Short Laser Pulse

    NASA Astrophysics Data System (ADS)

    Aglitskiy, Y.; Karasik, M.; Velikovich, A. L.; Serlin, V.; Weaver, J. L.; Kessler, T. J.; Schmitt, A. J.; Obenschain, S. P.; Metzler, N.; Oh, J.

    2010-11-01

    When a short (sub-ns) laser pulse deposits finite energy in a target, the shock wave launched into it is immediately followed by a rarefaction wave. If the irradiated surface is rippled, theory and simulations predict strong oscillations of the areal mass perturbation amplitude in the target [A. L. Velikovich et al., Phys. Plasmas 10, 3270 (2003).] The first experiment designed to observe this effect has become possible by adding short-driving-pulse capability to the Nike laser, and has been scheduled for the fall of 2010. Simulations show that while the driving pulse of 0.3 ns is on, the areal mass perturbation amplitude grows by a factor ˜2 due to ablative Richtmyer-Meshkov instability. It then decreases, reverses phase, and reaches another maximum, also about twice its initial value, shortly after the shock breakout at the rear target surface. This signature behavior is observable with the monochromatic x-ray imaging diagnostics fielded on Nike.

  20. Cryosurgery with pulsed electric fields.

    PubMed

    Daniels, Charlotte S; Rubinsky, Boris

    2011-01-01

    This study explores the hypothesis that combining the minimally invasive surgical techniques of cryosurgery and pulsed electric fields will eliminate some of the major disadvantages of these techniques while retaining their advantages. Cryosurgery, tissue ablation by freezing, is a well-established minimally invasive surgical technique. One disadvantage of cryosurgery concerns the mechanism of cell death; cells at high subzero temperature on the outer rim of the frozen lesion can survive. Pulsed electric fields (PEF) are another minimally invasive surgical technique in which high strength and very rapid electric pulses are delivered across cells to permeabilize the cell membrane for applications such as gene delivery, electrochemotherapy and irreversible electroporation. The very short time scale of the electric pulses is disadvantageous because it does not facilitate real time control over the procedure. We hypothesize that applying the electric pulses during the cryosurgical procedure in such a way that the electric field vector is parallel to the heat flux vector will have the effect of confining the electric fields to the frozen/cold region of tissue, thereby ablating the cells that survive freezing while facilitating controlled use of the PEF in the cold confined region. A finite element analysis of the electric field and heat conduction equations during simultaneous tissue treatment with cryosurgery and PEF (cryosurgery/PEF) was used to study the effect of tissue freezing on electric fields. The study yielded motivating results. Because of decreased electrical conductivity in the frozen/cooled tissue, it experienced temperature induced magnified electric fields in comparison to PEF delivered to the unfrozen tissue control. This suggests that freezing/cooling confines and magnifies the electric fields to those regions; a targeting capability unattainable in traditional PEF. This analysis shows how temperature induced magnified and focused PEFs could be used to

  1. Cryosurgery with Pulsed Electric Fields

    PubMed Central

    Daniels, Charlotte S.; Rubinsky, Boris

    2011-01-01

    This study explores the hypothesis that combining the minimally invasive surgical techniques of cryosurgery and pulsed electric fields will eliminate some of the major disadvantages of these techniques while retaining their advantages. Cryosurgery, tissue ablation by freezing, is a well-established minimally invasive surgical technique. One disadvantage of cryosurgery concerns the mechanism of cell death; cells at high subzero temperature on the outer rim of the frozen lesion can survive. Pulsed electric fields (PEF) are another minimally invasive surgical technique in which high strength and very rapid electric pulses are delivered across cells to permeabilize the cell membrane for applications such as gene delivery, electrochemotherapy and irreversible electroporation. The very short time scale of the electric pulses is disadvantageous because it does not facilitate real time control over the procedure. We hypothesize that applying the electric pulses during the cryosurgical procedure in such a way that the electric field vector is parallel to the heat flux vector will have the effect of confining the electric fields to the frozen/cold region of tissue, thereby ablating the cells that survive freezing while facilitating controlled use of the PEF in the cold confined region. A finite element analysis of the electric field and heat conduction equations during simultaneous tissue treatment with cryosurgery and PEF (cryosurgery/PEF) was used to study the effect of tissue freezing on electric fields. The study yielded motivating results. Because of decreased electrical conductivity in the frozen/cooled tissue, it experienced temperature induced magnified electric fields in comparison to PEF delivered to the unfrozen tissue control. This suggests that freezing/cooling confines and magnifies the electric fields to those regions; a targeting capability unattainable in traditional PEF. This analysis shows how temperature induced magnified and focused PEFs could be used to

  2. Assessment of the electrochemical effects of pulsed electric fields in a biological cell suspension.

    PubMed

    Chafai, Djamel Eddine; Mehle, Andraž; Tilmatine, Amar; Maouche, Bachir; Miklavčič, Damijan

    2015-12-01

    Electroporation of cells is successfully used in biology, biotechnology and medicine. Practical problems still arise in the electroporation of cells in suspension. For example, the determination of cell electroporation is still a demanding and time-consuming task. Electric pulses also cause contamination of the solution by the metal released from the electrodes and create local enhancements of the electric field, leading to the occurrence of electrochemical reactions at the electrode/electrolyte interface. In our study, we investigated the possibility of assessing modifications to the cell environment caused by pulsed electric fields using electrochemical impedance spectroscopy. We designed an experimental protocol to elucidate the mechanism by which a pulsed electric field affects the electrode state in relation to different electrolyte conductivities at the interface. The results show that a pulsed electric field affects electrodes and its degree depends on the electrolyte conductivity. Evolution of the electrochemical reaction rate depends on the initial free charges and those generated by the pulsed electric field. In the presence of biological cells, the initial free charges in the medium are reduced. The electrical current path at low frequency is longer, i.e., conductivity is decreased, even in the presence of increased permeability of the cell membrane created by the pulsed electric field. Copyright © 2015 Elsevier B.V. All rights reserved.

  3. Synchrotron emission from nanowire array targets irradiated by ultraintense laser pulses

    NASA Astrophysics Data System (ADS)

    Martinez, B.; d’Humières, E.; Gremillet, L.

    2018-07-01

    We present a numerical study, based on two-dimensional particle-in-cell simulations, of the synchrotron emission induced during the interaction of femtosecond laser pulses of intensities I = 1021–1023 W cm‑2 with nanowire arrays. Through an extensive parametric scan on the target parameters, we identify and characterize several dominant radiation mechanisms, mainly depending on the transparency or opacity of the plasma produced by the wire expansion. At I = 1022 W m‑2, the emission of high-energy (>10 keV) photons attains a maximum conversion efficiency of ∼10% for 36–50 nm wire widths and 1 μm interspacing. This maximum radiation yield is found to be similar to that achieved in a uniform plasma of same average (sub-solid) density, but nanowire arrays provide efficient radiation sources over a broader parameter range. Moreover, we examine the variations of the photon spectra with the laser intensity and the wire material, and we demonstrate that the radiation efficiency can be further enhanced by adding a plasma mirror at the backside of the nanowire array. Finally, we briefly consider the influence of a finite laser focal spot and oblique incidence angle.

  4. Pulsed laser interactions with space debris: Target shape effects

    DOE PAGES

    Liedahl, D. A.; Rubenchik, A.; Libby, S. B.; ...

    2013-05-24

    Among the approaches to the proposed mitigation and remediation of the space debris problem is the de-orbiting of objects in low Earth orbit through irradiation by ground-based high-intensity pulsed lasers. Laser ablation of a thin surface layer causes target recoil, resulting in the depletion of orbital angular momentum and accelerated atmospheric re-entry. However, both the magnitude and direction of the recoil are shape dependent, a feature of the laser-based remediation concept that has received little attention. Since the development of a predictive capability is desirable, we have investigated the dynamical response to ablation of objects comprising a variety of shapes.more » We derive and demonstrate a simple analytical technique for calculating the ablation-driven transfer of linear momentum, emphasizing cases for which the recoil is not exclusively parallel to the incident beam. For the purposes of comparison and contrast, we examine one case of momentum transfer in the low-intensity regime, where photon pressure is the dominant momentum transfer mechanism, showing that shape and orientation effects influence the target response in a similar, but not identical, manner. As a result, we address the related problem of target spin and, by way of a few simple examples, show how ablation can alter the spin state of a target, which often has a pronounced effect on the recoil dynamics.« less

  5. Pulsed laser interactions with space debris: Target shape effects

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    Liedahl, D. A.; Rubenchik, A.; Libby, S. B.

    Among the approaches to the proposed mitigation and remediation of the space debris problem is the de-orbiting of objects in low Earth orbit through irradiation by ground-based high-intensity pulsed lasers. Laser ablation of a thin surface layer causes target recoil, resulting in the depletion of orbital angular momentum and accelerated atmospheric re-entry. However, both the magnitude and direction of the recoil are shape dependent, a feature of the laser-based remediation concept that has received little attention. Since the development of a predictive capability is desirable, we have investigated the dynamical response to ablation of objects comprising a variety of shapes.more » We derive and demonstrate a simple analytical technique for calculating the ablation-driven transfer of linear momentum, emphasizing cases for which the recoil is not exclusively parallel to the incident beam. For the purposes of comparison and contrast, we examine one case of momentum transfer in the low-intensity regime, where photon pressure is the dominant momentum transfer mechanism, showing that shape and orientation effects influence the target response in a similar, but not identical, manner. As a result, we address the related problem of target spin and, by way of a few simple examples, show how ablation can alter the spin state of a target, which often has a pronounced effect on the recoil dynamics.« less

  6. Nanosecond pulsed electric fields and the cell cycle

    NASA Astrophysics Data System (ADS)

    Mahlke, Megan A.

    Exposure to nanosecond pulsed electrical fields (nsPEFs) can cause poration of external and internal cell membranes, DNA damage, and disassociation of cytoskeletal components, all of which are capable of disrupting a cell's ability to replicate. The phase of the cell cycle at the time of exposure is linked to differential sensitivities to nsPEFs across cell lines, as DNA structure, membrane elasticity, and cytoskeletal structure change dramatically during the cell cycle. Additionally, nsPEFs are capable of activating cell cycle checkpoints, which could lead to apoptosis or slow population growth. NsPEFs are emerging as a method for treating tumors via apoptotic induction; therefore, investigating the relevance of nsPEFs and the cell cycle could translate into improved efficacy in tumor treatment. Populations of Jurkat and Chinese Hamster Ovary (CHO) cells were examined post-exposure (10 ns pulse trains at 150kV/cm) by analysis of DNA content via propidium iodide staining and flow cytometric analysis at various time points (1, 6, and 12h post-exposure) to determine population distribution in cell cycle phases. Additionally, CHO and Jurkat cells were synchronized in G1/S and G2/M phases, pulsed, and analyzed to evaluate the role of cell cycle phase in survival of nsPEFs. CHO populations appeared similar to sham populations post-nsPEFs but exhibited arrest in the G1 phase at 6h after exposure. Jurkat cells exhibited increased cell death after nsPEFs compared to CHO cells but did not exhibit checkpoint arrest at any observed time point. The G1/S phase checkpoint is partially controlled by the action of p53; the lack of an active p53 response in Jurkat cells could contribute to their ability to pass this checkpoint and resist cell cycle arrest. Both cell lines exhibited increased sensitivity to nsPEFs in G2/M phase. Live imaging of CHO cells after nsPEF exposure supports the theory of G1/S phase arrest, as a reduced number of cells undergo mitosis within 24 h when

  7. Bright attosecond γ-ray pulses from nonlinear Compton scattering with laser-illuminated compound targets

    NASA Astrophysics Data System (ADS)

    Zhu, Xing-Long; Chen, Min; Yu, Tong-Pu; Weng, Su-Ming; Hu, Li-Xiang; McKenna, Paul; Sheng, Zheng-Ming

    2018-04-01

    Attosecond light sources have the potential to open up totally unexplored research avenues in ultrafast science. However, the photon energies achievable using existing generation schemes are limited to the keV range. Here, we propose and numerically demonstrate an all-optical mechanism for the generation of bright MeV attosecond γ-photon beams with desirable angular momentum. Using a circularly polarized Laguerre-Gaussian laser pulse focused onto a cone-foil target, dense attosecond bunches ( ≲ 170 as ) of electrons are produced. The electrons interact with the laser pulse which is reflected by a plasma mirror, producing ultra-brilliant (˜1023 photons/s/mm2/mrad2/0.1%BW) multi-MeV (Eγ,max > 30 MeV) isolated attosecond ( ≲ 260 as ) γ-ray pulse trains. Moreover, the angular momentum is transferred to γ-photon beams via nonlinear Compton scattering of ultra-intense tightly focused laser pulse by energetic electrons. Such a brilliant attosecond γ-photon source would provide the possibilities in attosecond nuclear science.

  8. Modeling of beam-target interaction during pulsed electron beam ablation of graphite: Case of melting

    NASA Astrophysics Data System (ADS)

    Ali, Muddassir; Henda, Redhouane

    2017-02-01

    A one-dimensional thermal model based on a two-stage heat conduction equation is employed to investigate the ablation of graphite target during nanosecond pulsed electron beam ablation. This comprehensive model accounts for the complex physical phenomena comprised of target heating, melting and vaporization upon irradiation with a polyenergetic electron beam. Melting and vaporization effects induced during ablation are taken into account by introducing moving phase boundaries. Phase transition induced during ablation is considered through the temperature dependent thermodynamic properties of graphite. The effect of electron beam efficiency, power density, and accelerating voltage on ablation is analyzed. For an electron beam operating at an accelerating voltage of 15 kV and efficiency of 0.6, the model findings show that the target surface temperature can reach up to 7500 K at the end of the pulse. The surface begins to melt within 25 ns from the pulse start. For the same process conditions, the estimated ablation depth and ablated mass per unit area are about 0.60 μm and 1.05 μg/mm2, respectively. Model results indicate that ablation takes place primarily in the regime of normal vaporization from the surface. The results obtained at an accelerating voltage of 15 kV and efficiency factor of 0.6 are satisfactorily in good accordance with available experimental data in the literature.

  9. Oxygen targeting in preterm infants using the Masimo SET Radical pulse oximeter

    PubMed Central

    Johnston, Ewen D; Boyle, Breidge; Juszczak, Ed; King, Andy; Brocklehurst, Peter; Stenson, Ben J

    2011-01-01

    Background A pretrial clinical improvement project for the BOOST-II UK trial of oxygen saturation targeting revealed an artefact affecting saturation profiles obtained from the Masimo Set Radical pulse oximeter. Methods Saturation was recorded every 10 s for up to 2 weeks in 176 oxygen dependent preterm infants in 35 UK and Irish neonatal units between August 2006 and April 2009 using Masimo SET Radical pulse oximeters. Frequency distributions of % time at each saturation were plotted. An artefact affecting the saturation distribution was found to be attributable to the oximeter's internal calibration algorithm. Revised software was installed and saturation distributions obtained were compared with four other current oximeters in paired studies. Results There was a reduction in saturation values of 87–90%. Values above 87% were elevated by up to 2%, giving a relative excess of higher values. The software revision eliminated this, improving the distribution of saturation values. In paired comparisons with four current commercially available oximeters, Masimo oximeters with the revised software returned similar saturation distributions. Conclusions A characteristic of the software algorithm reduces the frequency of saturations of 87–90% and increases the frequency of higher values returned by the Masimo SET Radical pulse oximeter. This effect, which remains within the recommended standards for accuracy, is removed by installing revised software (board firmware V4.8 or higher). Because this observation is likely to influence oxygen targeting, it should be considered in the analysis of the oxygen trial results to maximise their generalisability. PMID:21378398

  10. Cell Fragmentation and Permeabilization by a 1 ns Pulse Driven Triple-Point Electrode

    PubMed Central

    Li, Joy; Cho, Michael

    2018-01-01

    Ultrashort electric pulses (ns-ps) are useful in gaining understanding as to how pulsed electric fields act upon biological cells, but the electric field intensity to induce biological responses is typically higher than longer pulses and therefore a high voltage ultrashort pulse generator is required. To deliver 1 ns pulses with sufficient electric field but at a relatively low voltage, we used a glass-encapsulated tungsten wire triple-point electrode (TPE) at the interface among glass, tungsten wire, and water when it is immersed in water. A high electric field (2 MV/cm) can be created when pulses are applied. However, such a high electric field was found to cause bubble emission and temperature rise in the water near the electrode. They can be attributed to Joule heating near the electrode. Adherent cells on a cover slip treated by the combination of these stimuli showed two major effects: (1) cells in a crater (<100 μm from electrode) were fragmented and the debris was blown away. The principal mechanism for the damage is presumed to be shear forces due to bubble collapse; and (2) cells in the periphery of the crater were permeabilized, which was due to the combination of bubble movement and microstreaming as well as pulsed electric fields. These results show that ultrashort electric fields assisted by microbubbles can cause significant cell response and therefore a triple-point electrode is a useful ablation tool for applications that require submillimeter precision. PMID:29744357

  11. Short infrared laser pulses increase cell membrane fluidity

    NASA Astrophysics Data System (ADS)

    Walsh, Alex J.; Cantu, Jody C.; Ibey, Bennett L.; Beier, Hope T.

    2017-02-01

    Short infrared laser pulses induce a variety of effects in cells and tissues, including neural stimulation and inhibition. However, the mechanism behind these physiological effects is poorly understood. It is known that the fast thermal gradient induced by the infrared light is necessary for these biological effects. Therefore, this study tests the hypothesis that the fast thermal gradient induced in a cell by infrared light exposure causes a change in the membrane fluidity. To test this hypothesis, we used the membrane fluidity dye, di-4-ANEPPDHQ, to investigate membrane fluidity changes following infrared light exposure. Di-4-ANEPPDHQ fluorescence was imaged on a wide-field fluorescence imaging system with dual channel emission detection. The dual channel imaging allowed imaging of emitted fluorescence at wavelengths longer and shorter than 647 nm for ratiometric assessment and computation of a membrane generalized polarization (GP) value. Results in CHO cells show increased membrane fluidity with infrared light pulse exposure and this increased fluidity scales with infrared irradiance. Full recovery of pre-infrared exposure membrane fluidity was observed. Altogether, these results demonstrate that infrared light induces a thermal gradient in cells that changes membrane fluidity.

  12. Solar cells utilizing pulsed-energy crystallized microcrystalline/polycrystalline silicon

    DOEpatents

    Kaschmitter, J.L.; Sigmon, T.W.

    1995-10-10

    A process for producing multi-terminal devices such as solar cells wherein a pulsed high energy source is used to melt and crystallize amorphous silicon deposited on a substrate which is intolerant to high processing temperatures, whereby the amorphous silicon is converted into a microcrystalline/polycrystalline phase. Dopant and hydrogenation can be added during the fabrication process which provides for fabrication of extremely planar, ultra shallow contacts which results in reduction of non-current collecting contact volume. The use of the pulsed energy beams results in the ability to fabricate high efficiency microcrystalline/polycrystalline solar cells on the so-called low-temperature, inexpensive plastic substrates which are intolerant to high processing temperatures.

  13. Solar cells utilizing pulsed-energy crystallized microcrystalline/polycrystalline silicon

    DOEpatents

    Kaschmitter, James L.; Sigmon, Thomas W.

    1995-01-01

    A process for producing multi-terminal devices such as solar cells wherein a pulsed high energy source is used to melt and crystallize amorphous silicon deposited on a substrate which is intolerant to high processing temperatures, whereby to amorphous silicon is converted into a microcrystalline/polycrystalline phase. Dopant and hydrogenization can be added during the fabrication process which provides for fabrication of extremely planar, ultra shallow contacts which results in reduction of non-current collecting contact volume. The use of the pulsed energy beams results in the ability to fabricate high efficiency microcrystalline/polycrystalline solar cells on the so-called low-temperature, inexpensive plastic substrates which are intolerant to high processing temperatures.

  14. An ex vivo feasibility experimental study on targeted cell surgery by high intensity focused ultrasound

    NASA Astrophysics Data System (ADS)

    Wang, Zhi Biao; Wu, Junru; Fang, Liao Qiong; Wang, Hua; Li, Fa Qi; Tian, Yun Bo; Gong, Xiao Bo; Zhang, Hong; Zhang, Lian; Feng, Ruo

    2012-10-01

    High intensity focused ultrasound (HIFU) has become a new noninvasive surgical modality in medicine. A portion of tissue seated inside a patient's body may experience coagulative necrosis after a few seconds of insonification by high intensity focused ultrasound (US) generated by an extracorporeal focusing US transducer. The region of tissue affected by coagulative necrosis (CN) usually has an ellipsoidal shape when the thermal effect due to US absorption plays the dominant role. Its long and short axes are parallel and perpendicular to the US propagation direction respectively. It was shown by ex vivo experiments that the dimension of the short and long axes of the tissue which experiences CN can be as small as 50 μm and 250 μm respectively after one second exposure of US pulse (the spatial and pulse average acoustic power is on the order of tens of Watts and the local acoustic spatial and temporal pulse averaged intensity is on the order of 3 × 104 W/cm2) generated by a 1.6 MHz HIFU transducer of 12 cm diameter and 11 cm geometric focal length (f-number = 0.92). The numbers of cells which suffered CN were estimated to be on the order of 40. This result suggests that HIFU is able to interact with tens of cells at/near its focal zone while keeping the neighboring cells minimally affected, and thus the targeted cell surgery may be achievable.

  15. High field terahertz pulse generation from plasma wakefield driven by tailored laser pulses

    NASA Astrophysics Data System (ADS)

    Chen, Zi-Yu

    2013-06-01

    A scheme to generate high field terahertz (THz) pulses by using tailored laser pulses interaction with a gas target is proposed. The laser wakefield based THz source is emitted from the asymmetric laser shape induced plasma transverse transient net currents. Particle-in-cell simulations show that THz emission with electric filed strength over 1 GV/cm can be obtained with incident laser at 1×1019 W/cm2 level, and the corresponding energy conversion efficiency is more than 10-4. The intensity scaling holds up to high field strengths. Such a source also has a broad tunability range in amplitude, frequency spectra, and temporal shape.

  16. Silicon solar cells by ion implantation and pulsed energy processing

    NASA Technical Reports Server (NTRS)

    Kirkpatrick, A. R.; Minnucci, J. A.; Shaughnessy, T. S.; Greenwald, A. C.

    1976-01-01

    A new method for fabrication of silicon solar cells is being developed around ion implantation in conjunction with pulsed electron beam techniques to replace conventional furnace processing. Solar cells can be fabricated totally in a vacuum environment at room temperature. Cells with 10% AM0 efficiency have been demonstrated. High efficiency cells and effective automated processing capabilities are anticipated.

  17. Conceptual moderator studies for the Spallation Neutron Source short-pulse second target station

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    Gallmeier, F. X., E-mail: gallmeierfz@ornl.gov; Lu, W.; Riemer, B. W.

    Candidate moderator configurations for a short-pulse second target station (STS) at the Oak Ridge National Laboratory Spallation Neutron Source (SNS) have been identified using a global optimizer framework built around the MCNPX particle transport code. Neutron brightness metrics were selected as the figure-of-merit. We assumed that STS would use one out of six proton pulses produced by an SNS accelerator upgraded to operate at 1.3 GeV proton energy, 2.8 MW power and 60 Hz repetition rate. The simulations indicate that the peak brightness can be increased by a factor of 5 and 2.5 on a per proton pulse basis comparedmore » to the SNS first target station for both coupled and decoupled para-hydrogen moderators, respectively. Additional increases by factors of 3 and 2 were demonstrated for coupled and decoupled moderators, respectively, by reducing the area of neutron emission from 100 × 100 mm{sup 2} to 20 × 20 mm{sup 2}. This increase in brightness has the potential to translate to an increase of beam intensity at the instruments’ sample positions even though the total neutron emission of the smaller moderator is less than that of the larger. This is especially true for instruments with small samples (beam dimensions). The increased fluxes in the STS moderators come at accelerated poison and de-coupler burnout and higher radiation-induced material damage rates per unit power, which overall translate into lower moderator lifetimes. A first effort was undertaken to group decoupled moderators into a cluster collectively positioning them at the peak neutron production zone in the target and having a three-port neutron emission scheme that complements that of a cylindrical coupled moderator.« less

  18. Observation of ionization shifts in K-shell emission from short-pulse laser irradiated micro-dot targets

    NASA Astrophysics Data System (ADS)

    Neumayer, Paul; Kritcher, Andrea; Landen, Otto; Lee, Haeja; Offerman, Dustin; Shipton, Eric; Glenzer, Siegfried

    2006-10-01

    X-ray Thomson scattering using short pulse laser generated intense line radiation has a great potential as a time-resolved temperature and density diagnostic for high-energy density states of matter. We present recent results characterizing Chlorine K-alpha and K-beta line emission obtained by irradiating Saran foil with 50 Terawatt laser pulses from the Callisto laser (Jupiter Laser Facility, Lawrence Livermore National Laboratory). Spectra from front and rear side emission are recorded simultaneously with high resolution HOPG spectrometers employing imaging plate detectors. Conversion efficiencies of laser pulse energy into x-ray line emission of several 10-5 are achieved and are maintained throughout up to 7 J of laser energy, thus constituting a short pulsed narrow band x-ray source of more than 10^11 photons. When the target size is reduced to 50 micrometer (``micro-dot'') a significant blue-shift of up to 5 eV is clearly observed. This can be attributed to higher ionization states of the target atoms indicating achievement of a high-temperature solid density state. This work was performed under the auspices of the U.S. Department of Energy by the Lawrence Livermore National Laboratory under Contract No. W-7405-ENG-48 and LDRD 05-ERI-003.

  19. Mechanical Strains Induced in Osteoblasts by Use of Point Femtosecond Laser Targeting

    PubMed Central

    Bomzon, Ze'ev; Day, Daniel; Gu, Min; Cartmell, Sarah

    2006-01-01

    A study demonstrating how ultrafast laser radiation stimulates osteoblasts is presented. The study employed a custom made optical system that allowed for simultaneous confocal cell imaging and targeted femtosecond pulse laser irradiation. When femtosecond laser light was focused onto a single cell, a rise in intracellular Ca2+ levels was observed followed by contraction of the targeted cell. This contraction caused deformation of neighbouring cells leading to a heterogeneous strain field throughout the monolayer. Quantification of the strain fields in the monolayer using digital image correlation revealed local strains much higher than threshold values typically reported to stimulate extracellular bone matrix production in vitro. This use of point targeting with femtosecond pulse lasers could provide a new method for stimulating cell activity in orthopaedic tissue engineering. PMID:23165014

  20. Pulse Width Dependence Of Pigment Cell Damage At 694 nm In Guinea Pig Skin

    NASA Astrophysics Data System (ADS)

    Dover, Jeffrey S.; Polla, Luigi L.; Margolis, Randall J.; Whitaker, Diana; Watanabe, Schinichi; Murphy, George F.; Parrish, John A.; Anderson, R. R.

    1987-03-01

    351 nm, 20-nsec XeF excimer laser irradiation has previously been shown to selectively target and damage melanosomes in human skin. In the following studies selective targeting with melanosomal photodisruption has been demonstrated in pigmented guinea pig skin with a Q-switched 40-nsec ruby laser, and a 750-nsec pulsed dye laser but not with a 400-usec pulsed dye laser. The pulse width dependence of melanosomal disruption, occurring only at pulsewidths shorter than the thermal relaxation time of the melanosome (0.5 - 1.0 usec), is in accordance with the theory of selective photothermolysis. Possible mechanisms of melanosomal photodisruption include development of sudden thermal gradients leading to cavitation or shock wave production.

  1. Correlation between electric field pulse induced long-lived permeabilization and fusogenicity in cell membranes.

    PubMed Central

    Teissié, J; Ramos, C

    1998-01-01

    Electric field pulses have been reported to induce long-lived permeabilization and fusogenicity on cell membranes. The two membrane property alterations are under the control of the field strength, the pulse duration, and the number of pulses. Experiments on mammalian cells pulsed by square wave form pulses and then brought into contact randomly through centrifugation revealed an even stronger analogy between the two processes. Permeabilization was known to affect well-defined regions of the cell surface. Fusion can be obtained only when permeabilized surfaces on the two partners were brought into contact. Permeabilization was under the control of the pulse duration and of the number of pulses. A similar relationship was observed as far as fusion is concerned. But a critical level of local permeabilization must be present for fusion to take place when contacts are created. The same conclusions are obtained from previous experiments on ghosts subjected to exponentially decaying field pulses and then brought into contact by dielectrophoresis. These observations are in agreement with a model of membrane fusion in which the merging of local random defects occurs when the two membranes are brought into contact. The local defects are considered part of the structural membrane reorganization induced by the external field. Their density is dependent on the pulse duration and number of pulses. They support the long-lived permeabilization. Their number must be very large to support the occurrence of membrane fusion. PMID:9545050

  2. Correlation between electric field pulse induced long-lived permeabilization and fusogenicity in cell membranes.

    PubMed

    Teissié, J; Ramos, C

    1998-04-01

    Electric field pulses have been reported to induce long-lived permeabilization and fusogenicity on cell membranes. The two membrane property alterations are under the control of the field strength, the pulse duration, and the number of pulses. Experiments on mammalian cells pulsed by square wave form pulses and then brought into contact randomly through centrifugation revealed an even stronger analogy between the two processes. Permeabilization was known to affect well-defined regions of the cell surface. Fusion can be obtained only when permeabilized surfaces on the two partners were brought into contact. Permeabilization was under the control of the pulse duration and of the number of pulses. A similar relationship was observed as far as fusion is concerned. But a critical level of local permeabilization must be present for fusion to take place when contacts are created. The same conclusions are obtained from previous experiments on ghosts subjected to exponentially decaying field pulses and then brought into contact by dielectrophoresis. These observations are in agreement with a model of membrane fusion in which the merging of local random defects occurs when the two membranes are brought into contact. The local defects are considered part of the structural membrane reorganization induced by the external field. Their density is dependent on the pulse duration and number of pulses. They support the long-lived permeabilization. Their number must be very large to support the occurrence of membrane fusion.

  3. Applications of the pulsed gas stripper technique at the GSI UNILAC

    NASA Astrophysics Data System (ADS)

    Scharrer, P.; Barth, W.; Bevcic, M.; Düllmann, Ch. E.; Gerhard, P.; Groening, L.; Horn, K. P.; Jäger, E.; Khuyagbaatar, J.; Krier, J.; Vormann, H.; Yakushev, A.

    2017-08-01

    In the frame of an upgrade program for the GSI UNILAC, preparing it for the use as an injector system for FAIR, a pulsed gas stripper cell was developed. It utilizes the required low duty cycle by applying a pulsed gas injection instead of a continuous gas inlet. The resulting lower gas consumption rate enables the use of low-Z gas targets over a wide range of stripper target thicknesses. The setup enables an increased flexibility for the accelerator by allowing the gas stripper to be used in time-sharing beam operation matching the capabilities of the GSI UNILAC like the acceleration of different ion beams in quasi-parallel operation. Measured charge state distributions of 238U, 50Ti, and CH3 beams on H2 and N2 gas highlight the benefits of the pulsed gas stripper cell for the accelerator operation and performance.

  4. INTERACTION OF LASER RADIATION WITH MATTER: Influence of a target on operation of a pulsed CO2 laser emitting microsecond pulses

    NASA Astrophysics Data System (ADS)

    Baranov, V. Yu; Dolgov, V. A.; Malyuta, D. D.; Mezhevov, V. S.; Semak, V. V.

    1987-12-01

    The profile of pulses emitted by a TEA CO2 laser with an unstable resonator changed as a result of interaction of laser radiation with the surface of a metal in the presence of a breakdown plasma. This influence of a target on laser operation and its possible applications in laser processing of materials are analyzed.

  5. Nanosecond laser pulse stimulation of spiral ganglion neurons and model cells.

    PubMed

    Rettenmaier, Alexander; Lenarz, Thomas; Reuter, Günter

    2014-04-01

    Optical stimulation of the inner ear has recently attracted attention, suggesting a higher frequency resolution compared to electrical cochlear implants due to its high spatial stimulation selectivity. Although the feasibility of the effect is shown in multiple in vivo experiments, the stimulation mechanism remains open to discussion. Here we investigate in single-cell measurements the reaction of spiral ganglion neurons and model cells to irradiation with a nanosecond-pulsed laser beam over a broad wavelength range from 420 nm up to 1950 nm using the patch clamp technique. Cell reactions were wavelength- and pulse-energy-dependent but too small to elicit action potentials in the investigated spiral ganglion neurons. As the applied radiant exposure was much higher than the reported threshold for in vivo experiments in the same laser regime, we conclude that in a stimulation paradigm with nanosecond-pulses, direct neuronal stimulation is not the main cause of optical cochlea stimulation.

  6. Effect of positive pulse charge waveforms on the energy efficiency of lead-acid traction cells

    NASA Technical Reports Server (NTRS)

    Smithrick, J. J.

    1981-01-01

    The effects of four different charge methods on the energy conversion efficiency of 300 ampere hour lead acid traction cells were investigated. Three of the methods were positive pulse charge waveforms; the fourth, a constant current method, was used as a baseline of comparison. The positive pulse charge waveforms were: 120 Hz full wave rectified sinusoidal; 120 Hz silicon controlled rectified; and 1 kHz square wave. The constant current charger was set at the time average pulse current of each pulse waveform, which was 150 amps. The energy efficiency does not include charger losses. The lead acid traction cells were charged to 70 percent of rated ampere hour capacity in each case. The results of charging the cells using the three different pulse charge waveforms indicate there was no significant difference in energy conversion efficiency when compared to constant current charging at the time average pulse current value.

  7. The influence of femtosecond laser pulse wavelength on embryonic stem cell differentiation

    NASA Astrophysics Data System (ADS)

    Mthunzi, Patience

    2012-10-01

    Stem cells are rich in proteins, carbohydrates, deoxyribonucleic acid (DNA), ribonucleic acid (RNA) and various other cellular components which are responsible for a diversity of functions. Mostly the building blocks of these intracellular entities play an active role in absorbing ultra-violet (UV) and visible light sources. Light-matter interactions in biomaterials are a complex situation and subsequent damage may not always amount only from wavelength dependent effects but may also be driven by a wealth of other optical parameters which may lead to a variety photochemical reactions. Previously, literature has reported efficient photo-transfection and differentiation of pluripotent stem cells via near infrared (NIR) femtosecond (fs) laser pulses with minimum compromise to their viability. Therefore, in this study the influence of using different fs laser wavelengths on optical stem cell transfection and differentiation is investigated. A potassium titanyl phosphate (KTP) crystal was employed in frequency doubling a 1064 nm fs laser beam. The newly generated 532 nm fs pulsed beam was then utilized for the first time in transient photo-transfection of ES-E14TG2a mouse embryonic stem (mES) cells. Compared to using 1064 nm fs pulses which non-invasively introduce plasmid DNA and other macromolecules into mES cells, our results showed a significant decline in the photo-transfection efficiency following transfecting with a pulsed fs visible green beam.

  8. Targeting B Cells and Plasma Cells in Autoimmune Diseases

    PubMed Central

    Hofmann, Katharina; Clauder, Ann-Katrin; Manz, Rudolf Armin

    2018-01-01

    Success with B cell depletion using rituximab has proven the concept that B lineage cells represent a valid target for the treatment of autoimmune diseases, and has promoted the development of other B cell targeting agents. Present data confirm that B cell depletion is beneficial in various autoimmune disorders and also show that it can worsen the disease course in some patients. These findings suggest that B lineage cells not only produce pathogenic autoantibodies, but also significantly contribute to the regulation of inflammation. In this review, we will discuss the multiple pro- and anti-inflammatory roles of B lineage cells play in autoimmune diseases, in the context of recent findings using B lineage targeting therapies. PMID:29740441

  9. Investigation of the LMJ ignition target sensitivity to the laser pulse shape with 2D integrated calculations

    NASA Astrophysics Data System (ADS)

    Cherfils, Catherine; Malinie, Guy; Boniface, Claude; Gauthier, Pascal; Laffite, Stephane; Loiseau, Pascal

    2010-11-01

    The A943 cryogenic target in a Rugby hohlraum is our current nominal design for ignition with 160 beams on the Laser MegaJoule (Laffite et al 2007, 49th Annual Meeting of the Division of Plasma Physics, Loiseau et al 2010, 40th Annual Anomalous Absorption Conference). In this study we redesign the laser pulse of the target under the form of a sum of six supergaussians, which is more amenable to a sensitivity study : four supergaussians are used to launch the four main shocks in the capsule, and two additional supergaussians are used first to remove the LEH windows and then to control the acceleration of the first shock, respectively. We use our 2D FCI2 code to compare the radiation hydro of the capsule, obtained with this new pulse, to what was previously obtained. We investigate the sensitivity of the yield on some parameters, which are the maximum powers and respective timings of the different components of the laser pulse.

  10. Expression of a Broad Array of Negative Costimulatory Molecules and Blimp-1 in T Cells following Priming by HIV-1 Pulsed Dendritic Cells

    PubMed Central

    Shankar, Esaki Muthu; Che, Karlhans Fru; Messmer, Davorka; Lifson, Jeffrey D; Larsson, Marie

    2011-01-01

    Accumulating evidence indicates that immune impairment in persistent viral infections could lead to T-cell exhaustion. To evaluate the potential contribution of induction of negative costimulatory molecules to impaired T-cell responses, we primed naïve T cells with mature monocyte-derived dendritic cells (MDDCs) pulsed with HIV-1 in vitro. We used quantitative real-time polymerase chain reaction and flow cytometry, respectively, to compare the gene and surface-protein expression profiles of naïve T cells primed with HIV-pulsed or mock-pulsed DCs. We detected elevated expressions of negative costimulatory molecules, including lymphocyte activation gene-3 (LAG-3), CD160, cytolytic T-lymphocyte antigen-4 (CTLA-4), T-cell immunoglobulin mucin-containing domain-3 (TIM-3), programmed death-1 (PD-1) and TRAIL (tumor necrosis-factor–related apoptosis-inducing ligand) in T cells primed by HIV-pulsed DCs. The PD-1+ T-cell population also coexpressed TIM-3, LAG-3, and CTLA-4. Interestingly, we also found an increase in gene expression of the transcriptional repressors Blimp-1 (B-lymphocyte–induced maturation protein-1) and Foxp3 (forkhead transcription factor) in T-cells primed by HIV-pulsed DCs; Blimp-1 expression was directly proportional to the expression of the negative costimulatory molecules. Furthermore, levels of the effector cytokines interleukin-2, tumor necrosis factor-α and interferon-γ, and perforin and granzyme B were decreased in T-cell populations primed by HIV-pulsed DCs. In conclusion, in vitro priming of naïve T-cells with HIV-pulsed DC leads to expansion of T cells with coexpression of a broad array of negative costimulatory molecules and Blimp-1, with potential deleterious consequences for T-cell responses. PMID:21103670

  11. Piston cylinder cell for high pressure ultrasonic pulse echo measurements.

    PubMed

    Kepa, M W; Ridley, C J; Kamenev, K V; Huxley, A D

    2016-08-01

    Ultrasonic techniques such as pulse echo, vibrating reed, or resonant ultrasound spectroscopy are powerful probes not only for studying elasticity but also for investigating electronic and magnetic properties. Here, we report on the design of a high pressure ultrasonic pulse echo apparatus, based on a piston cylinder cell, with a simplified electronic setup that operates with a single coaxial cable and requires sample lengths of mm only. The design allows simultaneous measurements of ultrasonic velocities and attenuation coefficients up to a pressure of 1.5 GPa. We illustrate the performance of the cell by probing the phase diagram of a single crystal of the ferromagnetic superconductor UGe2.

  12. Piston cylinder cell for high pressure ultrasonic pulse echo measurements

    NASA Astrophysics Data System (ADS)

    Kepa, M. W.; Ridley, C. J.; Kamenev, K. V.; Huxley, A. D.

    2016-08-01

    Ultrasonic techniques such as pulse echo, vibrating reed, or resonant ultrasound spectroscopy are powerful probes not only for studying elasticity but also for investigating electronic and magnetic properties. Here, we report on the design of a high pressure ultrasonic pulse echo apparatus, based on a piston cylinder cell, with a simplified electronic setup that operates with a single coaxial cable and requires sample lengths of mm only. The design allows simultaneous measurements of ultrasonic velocities and attenuation coefficients up to a pressure of 1.5 GPa. We illustrate the performance of the cell by probing the phase diagram of a single crystal of the ferromagnetic superconductor UGe2.

  13. Target diagnostics for commissioning the AWE HELEN Laser Facility 100 TW chirped pulse amplification beam

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    Eagleton, R. T.; Clark, E. L.; Davies, H. M.

    2006-10-15

    The capability of the HELEN laser at the Atomic Weapons Establishment Aldermaston has been enhanced by the addition of a short-pulse laser beam to augment the twin opposing nanosecond time scale beams. The short-pulse beam utilizes the chirped pulse amplification (CPA) technique and is capable of delivering up to 60 J on target in a 500 fs pulse, around 100 TW, at the fundamental laser wavelength of 1.054 {mu}m. During the commissioning phase a number of diagnostic systems have been fielded, these include: x-ray pinhole imaging of the laser heated spot, charged particle time of flight, thermoluminescent dosimeter array, calibratedmore » radiochromic film, and CR39 nuclear track detector. These diagnostic systems have been used to verify the performance of the CPA beam to achieve a focused intensity of around 10{sup 19} W cm{sup -2} and to underwrite the facility radiological safety system.« less

  14. Pharmacologic suppression of target cell recognition by engineered T cells expressing chimeric T-cell receptors.

    PubMed

    Alvarez-Vallina, L; Yañez, R; Blanco, B; Gil, M; Russell, S J

    2000-04-01

    Adoptive therapy with autologous T cells expressing chimeric T-cell receptors (chTCRs) is of potential interest for the treatment of malignancy. To limit possible T-cell-mediated damage to normal tissues that weakly express the targeted tumor antigen (Ag), we have tested a strategy for the suppression of target cell recognition by engineered T cells. Jurkat T cells were transduced with an anti-hapten chTCR tinder the control of a tetracycline-suppressible promoter and were shown to respond to Ag-positive (hapten-coated) but not to Ag-negative target cells. The engineered T cells were then reacted with hapten-coated target cells at different effector to target cell ratios before and after exposure to tetracycline. When the engineered T cells were treated with tetracycline, expression of the chTCR was greatly decreased and recognition of the hapten-coated target cells was completely suppressed. Tetracycline-mediated suppression of target cell recognition by engineered T cells may be a useful strategy to limit the toxicity of the approach to cancer gene therapy.

  15. Spectral and temporal characteristics of target current and electromagnetic pulse induced by nanosecond laser ablation

    NASA Astrophysics Data System (ADS)

    Krása, J.; De Marco, M.; Cikhardt, J.; Pfeifer, M.; Velyhan, A.; Klír, D.; Řezáč, K.; Limpouch, J.; Krouský, E.; Dostál, J.; Ullschmied, J.; Dudžák, R.

    2017-06-01

    The current balancing the target charging and the emission of transient electromagnetic pulses (EMP) driven by the interaction of a focused 1.315 μm iodine 300 ps PALS laser with metallic and plastic targets were measured with the use of inductive probes. It is experimentally proven that the duration of return target currents and EMPs is much longer than the duration of laser-target interaction. The laser-produced plasma is active after the laser-target interaction. During this phase, the target acts as a virtual cathode and the plasma-target interface expands. A double exponential function is used in order to obtain the temporal characteristics of EMP. The rise time of EMPs fluctuates in the range up to a few tens of nanoseconds. Frequency spectra of EMP and target currents are modified by resonant frequencies of the interaction chamber.

  16. Generation of quasi-monoenergetic protons from a double-species target driven by the radiation pressure of an ultraintense laser pulse

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    Pae, Ki Hong; Kim, Chul Min, E-mail: chulmin@gist.ac.kr; Advanced Photonics Research Institute, Gwangju Institute of Science and Technology, Gwangju 61005

    In laser-driven proton acceleration, generation of quasi-monoenergetic proton beams has been considered a crucial feature of the radiation pressure acceleration (RPA) scheme, but the required difficult physical conditions have hampered its experimental realization. As a method to generate quasi-monoenergetic protons under experimentally viable conditions, we investigated using double-species targets of controlled composition ratio in order to make protons bunched in the phase space in the RPA scheme. From a modified optimum condition and three-dimensional particle-in-cell simulations, we showed by varying the ion composition ratio of proton and carbon that quasi-monoenergetic protons could be generated from ultrathin plane targets irradiated withmore » a circularly polarized Gaussian laser pulse. The proposed scheme should facilitate the experimental realization of ultrashort quasi-monoenergetic proton beams for unique applications in high field science.« less

  17. Frequency specificity in intercellular communication. Influence of patterns of periodic signaling on target cell responsiveness.

    PubMed Central

    Li, Y; Goldbeter, A

    1989-01-01

    Cells often communicate by means of periodic signals, as exemplified by a large number of hormones and by the aggregation of Dictyostelium discoideum amebas in response to periodic pulses of cyclic AMP. Periodic signaling allows bypassing the phenomenon of desensitization brought about by constant stimuli. To gain further insight into the efficiency of pulsatile signaling, we analyze the effect of periodic stimulation on the dynamic behavior of a receptor system capable of desensitization toward its ligand. We first show that the receptor system adapts to square-wave stimuli, i.e., the response eventually reaches a steady, periodic pattern after a transient phase. By analyzing the dependence of the response on the characteristics of the square-wave stimulation, we show that there exist a waveform and a period of that signal that result in maximum responsiveness of the target system. Similar results are obtained when the signal takes the more realistic form of a periodically repeated stimulation followed by exponential decay of the ligand. The results are discussed with respect to the role of pulsatile secretion of gonadotropin-releasing hormone (GnRH) by the hypothalamus and of periodic signaling by cyclic AMP pulses in Dictyostelium. The analysis accounts for the existence, in both cases, of an optimal frequency and waveform of the periodic stimulus that correspond to maximum target cell responsiveness. PMID:2930817

  18. Electron heating enhancement by frequency-chirped laser pulses

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    Yazdani, E.; Afarideh, H., E-mail: hafarideh@aut.ac.ir; Sadighi-Bonabi, R., E-mail: Sadighi@sharif.ir

    2014-09-14

    Propagation of a chirped laser pulse with a circular polarization through an uprising plasma density profile is studied by using 1D-3V particle-in-cell simulation. The laser penetration depth is increased in an overdense plasma compared to an unchirped pulse. The induced transparency due to the laser frequency chirp results in an enhanced heating of hot electrons as well as increased maximum longitudinal electrostatic field at the back side of the solid target, which is very essential in target normal sheath acceleration regime of proton acceleration. For an applied chirp parameter between 0.008 and 0.01, the maximum amount of the electrostatic fieldmore » is improved by a factor of 2. Furthermore, it is noticed that for a chirped laser pulse with a₀=5, because of increasing the plasma transparency length, the laser pulse can penetrate up to about n{sub e}≈6n{sub c}, where n{sub c} is plasma critical density. It shows 63% increase in the effective critical density compared to the relativistic induced transparency regime for an unchirped condition.« less

  19. Piston cylinder cell for high pressure ultrasonic pulse echo measurements

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    Kepa, M. W., E-mail: mkepa@staffmail.ed.ac.uk; Huxley, A. D.; Ridley, C. J.

    2016-08-15

    Ultrasonic techniques such as pulse echo, vibrating reed, or resonant ultrasound spectroscopy are powerful probes not only for studying elasticity but also for investigating electronic and magnetic properties. Here, we report on the design of a high pressure ultrasonic pulse echo apparatus, based on a piston cylinder cell, with a simplified electronic setup that operates with a single coaxial cable and requires sample lengths of mm only. The design allows simultaneous measurements of ultrasonic velocities and attenuation coefficients up to a pressure of 1.5 GPa. We illustrate the performance of the cell by probing the phase diagram of a singlemore » crystal of the ferromagnetic superconductor UGe{sub 2}.« less

  20. Human immune cell targeting of protein nanoparticles - caveospheres

    NASA Astrophysics Data System (ADS)

    Glass, Joshua J.; Yuen, Daniel; Rae, James; Johnston, Angus P. R.; Parton, Robert G.; Kent, Stephen J.; de Rose, Robert

    2016-04-01

    Nanotechnology has the power to transform vaccine and drug delivery through protection of payloads from both metabolism and off-target effects, while facilitating specific delivery of cargo to immune cells. However, evaluation of immune cell nanoparticle targeting is conventionally restricted to monocultured cell line models. We generated human caveolin-1 nanoparticles, termed caveospheres, which were efficiently functionalized with monoclonal antibodies. Using this platform, we investigated CD4+ T cell and CD20+ B cell targeting within physiological mixtures of primary human blood immune cells using flow cytometry, imaging flow cytometry and confocal microscopy. Antibody-functionalization enhanced caveosphere binding to targeted immune cells (6.6 to 43.9-fold) within mixed populations and in the presence of protein-containing fluids. Moreover, targeting caveospheres to CCR5 enabled caveosphere internalization by non-phagocytic CD4+ T cells--an important therapeutic target for HIV treatment. This efficient and flexible system of immune cell-targeted caveosphere nanoparticles holds promise for the development of advanced immunotherapeutics and vaccines.

  1. Energy transport in short-pulse-laser-heated targets measured using extreme ultraviolet laser backlighting.

    PubMed

    Wilson, L A; Tallents, G J; Pasley, J; Whittaker, D S; Rose, S J; Guilbaud, O; Cassou, K; Kazamias, S; Daboussi, S; Pittman, M; Delmas, O; Demailly, J; Neveu, O; Ros, D

    2012-08-01

    The accurate characterization of thermal electron transport and the determination of heating by suprathermal electrons in laser driven solid targets are both issues of great importance to the current experiments being performed at the National Ignition Facility, which aims to achieve thermonuclear fusion ignition using lasers. Ionization, induced by electronic heat conduction, can cause the opacity of a material to drop significantly once bound-free photoionization is no longer energetically possible. We show that this drop in opacity enables measurements of the transmission of extreme ultraviolet (EUV) laser pulses at 13.9 nm to act as a signature of the heating of thin (50 nm) iron layers with a 50-nm thick parylene-N (CH) overlay irradiated by 35-fs pulses at irradiance 3×10(16) Wcm(-2). Comparing EUV transmission measurements at different times after irradiation to fluid code simulations shows that the target is instantaneously heated by hot electrons (with approximately 10% of the laser energy), followed by thermal conduction with a flux limiter of ≈0.05.

  2. Nanosecond pulsed electric fields have differential effects on cells in the S-phase.

    PubMed

    Hall, Emily H; Schoenbach, Karl H; Beebe, Stephen J

    2007-03-01

    Nanosecond pulsed electric fields (nsPEFs) are a type of nonthermal, nonionizing radiation that exhibit intense electric fields with high power, but low energy. NsPEFs extend conventional electroporation (EP) to affect intracellular structures and functions and depending on the intensity, can induce lethal and nonlethal cell signaling. In this study, HCT116 human colon carcinoma cells were synchronized to the S-phase or remained unsynchronized, exposed to electric fields of 60 kV/cm with either 60-ns or 300-ns durations, and analyzed for apoptosis and proliferative markers. Several nsPEF structural and functional targets were identified. Unlike unsynchronized cells, S-phase cells under limiting conditions exhibited greater membrane integrity and caspase activation and maintained cytoskeletal structure. Regardless of synchronization, cells exposed to nsPEFs under these conditions primarily survived, but exhibited some turnover and delayed proliferation in cell populations, as well as reversible increases in phosphatidylserine externalization, membrane integrity, and nuclei size. These results show that nsPEFs can act as a nonligand agonist to modulate plasma membrane (PM) and intracellular structures and functions, as well as differentially affect cells in the S-phase, but without effect on cell survival. Furthermore, nsPEF effects on the nucleus and cytoskeleton may provide synergistic therapeutic actions with other agents, such as ionizing radiation or chemotherapeutics that affect these same structures.

  3. Der p 1-pulsed myeloid and plasmacytoid dendritic cells from house dust mite-sensitized allergic patients dysregulate the T cell response.

    PubMed

    Charbonnier, Anne-Sophie; Hammad, Hamida; Gosset, Philippe; Stewart, Geoffrey A; Alkan, Sefik; Tonnel, André-Bernard; Pestel, Joël

    2003-01-01

    Although reports suggest that dendritic cells (DC) are involved in the allergic reaction characterized by a T helper cell type 2 (Th2) profile, the role of myeloid (M-DC) and plasmacytoid DC (P-DC), controlling the balance Th1/Th2, remains unknown. Here, we showed that in Dermatophagoides pteronyssinus (Dpt)-sensitized allergic patients and in healthy donors, M-DC displayed a higher capacity to capture Der p 1, a major allergen of Dpt, than did P-DC. However, Der p 1-pulsed M-DC from healthy subjects overexpressed CD80 and secreted interleukin (IL)-10, whereas M-DC from allergic patients did not. In contrast, with Der p 1-pulsed P-DC from both groups, no increase in human leukocyte antigen-DR, CD80, and CD86 and no IL-10 secretion were detected. When cocultured with allogeneic naive CD4(+) T cells from healthy donors, Der p 1-pulsed M-DC from allergic patients favored a Th1 profile [interferon (IFN)-gamma(high)/IL-4(low)] and Der p 1-pulsed P-DC, a Th2 profile (IFN-gamma(low)/IL-4(high)). In healthy donors, no T cell polarization (IFN-gamma(low)/IL-4(low)) was induced by Der p 1-pulsed M-DC or P-DC, but in response to Der p 1-pulsed M-DC, T cells secreted IL-10. The neutralization of IL-10 produced by Der p 1-pulsed M-DC from healthy donors led to an inhibition of IL-10 production by T cells and a polarization toward a type 1. Thus, IL-10 produced by M-DC might be an essential mediator controlling the balance between tolerance and allergic status. In addition, P-DC could contribute to the steady state in healthy donors or to the development of a Th2 response in allergic donors.

  4. Aligned copper nanorod arrays for highly efficient generation of intense ultra-broadband THz pulses.

    PubMed

    Mondal, S; Wei, Q; Ding, W J; Hafez, H A; Fareed, M A; Laramée, A; Ropagnol, X; Zhang, G; Sun, S; Sheng, Z M; Zhang, J; Ozaki, T

    2017-01-10

    We demonstrate an intense broadband terahertz (THz) source based on the interaction of relativistic-intensity femtosecond lasers with aligned copper nanorod array targets. For copper nanorod targets with a length of 5 μm, a maximum 13.8 times enhancement in the THz pulse energy (in ≤20 THz spectral range) is measured as compared to that with a thick plane copper target under the same laser conditions. A further increase in the nanorod length leads to a decrease in the THz pulse energy at medium frequencies (≤20 THz) and increase of the electromagnetic pulse energy in the high-frequency range (from 20-200 THz). For the latter, we measure a maximum energy enhancement of 28 times for the nanorod targets with a length of 60 μm. Particle-in-cell simulations reveal that THz pulses are mostly generated by coherent transition radiation of laser produced hot electrons, which are efficiently enhanced with the use of nanorod targets. Good agreement is found between the simulation and experimental results.

  5. Aligned copper nanorod arrays for highly efficient generation of intense ultra-broadband THz pulses

    PubMed Central

    Mondal, S.; Wei, Q.; Ding, W. J.; Hafez, H. A.; Fareed, M. A.; Laramée, A.; Ropagnol, X.; Zhang, G.; Sun, S.; Sheng, Z. M.; Zhang, J.; Ozaki, T.

    2017-01-01

    We demonstrate an intense broadband terahertz (THz) source based on the interaction of relativistic-intensity femtosecond lasers with aligned copper nanorod array targets. For copper nanorod targets with a length of 5 μm, a maximum 13.8 times enhancement in the THz pulse energy (in ≤20 THz spectral range) is measured as compared to that with a thick plane copper target under the same laser conditions. A further increase in the nanorod length leads to a decrease in the THz pulse energy at medium frequencies (≤20 THz) and increase of the electromagnetic pulse energy in the high-frequency range (from 20–200 THz). For the latter, we measure a maximum energy enhancement of 28 times for the nanorod targets with a length of 60 μm. Particle-in-cell simulations reveal that THz pulses are mostly generated by coherent transition radiation of laser produced hot electrons, which are efficiently enhanced with the use of nanorod targets. Good agreement is found between the simulation and experimental results. PMID:28071764

  6. Aligned copper nanorod arrays for highly efficient generation of intense ultra-broadband THz pulses

    NASA Astrophysics Data System (ADS)

    Mondal, S.; Wei, Q.; Ding, W. J.; Hafez, H. A.; Fareed, M. A.; Laramée, A.; Ropagnol, X.; Zhang, G.; Sun, S.; Sheng, Z. M.; Zhang, J.; Ozaki, T.

    2017-01-01

    We demonstrate an intense broadband terahertz (THz) source based on the interaction of relativistic-intensity femtosecond lasers with aligned copper nanorod array targets. For copper nanorod targets with a length of 5 μm, a maximum 13.8 times enhancement in the THz pulse energy (in ≤20 THz spectral range) is measured as compared to that with a thick plane copper target under the same laser conditions. A further increase in the nanorod length leads to a decrease in the THz pulse energy at medium frequencies (≤20 THz) and increase of the electromagnetic pulse energy in the high-frequency range (from 20-200 THz). For the latter, we measure a maximum energy enhancement of 28 times for the nanorod targets with a length of 60 μm. Particle-in-cell simulations reveal that THz pulses are mostly generated by coherent transition radiation of laser produced hot electrons, which are efficiently enhanced with the use of nanorod targets. Good agreement is found between the simulation and experimental results.

  7. Time differentiated nuclear resonance spectroscopy coupled with pulsed laser heating in diamond anvil cells

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    Kupenko, I., E-mail: kupenko@esrf.fr; Strohm, C.; ESRF-The European Synchrotron, CS 40220, 38043 Grenoble Cedex 9

    2015-11-15

    Developments in pulsed laser heating applied to nuclear resonance techniques are presented together with their applications to studies of geophysically relevant materials. Continuous laser heating in diamond anvil cells is a widely used method to generate extreme temperatures at static high pressure conditions in order to study the structure and properties of materials found in deep planetary interiors. The pulsed laser heating technique has advantages over continuous heating, including prevention of the spreading of heated sample and/or the pressure medium and, thus, a better stability of the heating process. Time differentiated data acquisition coupled with pulsed laser heating in diamondmore » anvil cells was successfully tested at the Nuclear Resonance beamline (ID18) of the European Synchrotron Radiation Facility. We show examples applying the method to investigation of an assemblage containing ε-Fe, FeO, and Fe{sub 3}C using synchrotron Mössbauer source spectroscopy, FeCO{sub 3} using nuclear inelastic scattering, and Fe{sub 2}O{sub 3} using nuclear forward scattering. These examples demonstrate the applicability of pulsed laser heating in diamond anvil cells to spectroscopic techniques with long data acquisition times, because it enables stable pulsed heating with data collection at specific time intervals that are synchronized with laser pulses.« less

  8. Heating in short-pulse laser-driven cone-capped wire targets

    NASA Astrophysics Data System (ADS)

    Mason, R. J.; Wei, M.; King, J.; Beg, F.; Stephens, R. B.

    2007-11-01

    The 2-D implicit hybrid simulation code e-PLAS has been used to study heating in cone-capped copper wire targets. The code e-PLAS tracks collisional particle-in-cell (PIC) electrons traversing background plasma of collisional Eulerian cold electron and ion fluids. It computes E- and B-fields by the Implicit Moment Method [1,2]. In recent experiments [3] at the Vulcan laser facility, sub- picosecond laser pulses at 1.06 μm, and 4.0 x 10^20 W/cm^2 intensity were focused into thin-walled (˜10 μm) cones attached to copper wires. The wire diameter was varied from 10-40 μm with a typical length of 1 mm. We characterize heating of the wires as a function of their diameters and length, and relate modifications of this heating to changes in the assumed laser-generated hot electron spectrum and directivity. As in recent nail experiments [4], the cones can serve as reservoirs for hot electrons, diverting them from passage down the wires. [1] R. J. Mason, and C. Cranfill, IEEE Trans. Plasma Sci. PS-14, 45 (1986). [2] R. J. Mason, J. Comp. Phys. 71, 429 (1987). [3] J. King et al., to be submitted to Phys. Rev. Lett.. [4] R. J. Mason, M. Wei, F. Beg, R. Stephens, and C. Snell, in Proc. of ICOPS07, Albuquerque, NM, June 17-22, 2007, Talk 7D4.

  9. Rigor of cell fate decision by variable p53 pulses and roles of cooperative gene expression by p53

    PubMed Central

    Murakami, Yohei; Takada, Shoji

    2012-01-01

    Upon DNA damage, the cell fate decision between survival and apoptosis is largely regulated by p53-related networks. Recent experiments found a series of discrete p53 pulses in individual cells, which led to the hypothesis that the cell fate decision upon DNA damage is controlled by counting the number of p53 pulses. Under this hypothesis, Sun et al. (2009) modeled the Bax activation switch in the apoptosis signal transduction pathway that can rigorously “count” the number of uniform p53 pulses. Based on experimental evidence, here we use variable p53 pulses with Sun et al.’s model to investigate how the variability in p53 pulses affects the rigor of the cell fate decision by the pulse number. Our calculations showed that the experimentally anticipated variability in the pulse sizes reduces the rigor of the cell fate decision. In addition, we tested the roles of the cooperativity in PUMA expression by p53, finding that lower cooperativity is plausible for more rigorous cell fate decision. This is because the variability in the p53 pulse height is more amplified in PUMA expressions with more cooperative cases. PMID:27857606

  10. High energy protons generation by two sequential laser pulses

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    Wang, Xiaofeng; Shen, Baifei, E-mail: bfshen@mail.shcnc.ac.cn, E-mail: zhxm@siom.ac.cn; Zhang, Xiaomei, E-mail: bfshen@mail.shcnc.ac.cn, E-mail: zhxm@siom.ac.cn

    2015-04-15

    The sequential proton acceleration by two laser pulses of relativistic intensity is proposed to produce high energy protons. In the scheme, a relativistic super-Gaussian (SG) laser pulse followed by a Laguerre-Gaussian (LG) pulse irradiates dense plasma attached by underdense plasma. A proton beam is produced from the target and accelerated in the radiation pressure regime by the short SG pulse and then trapped and re-accelerated in a special bubble driven by the LG pulse in the underdense plasma. The advantages of radiation pressure acceleration and LG transverse structure are combined to achieve the effective trapping and acceleration of protons. Inmore » a two-dimensional particle-in-cell simulation, protons of 6.7 GeV are obtained from a 2 × 10{sup 22 }W/cm{sup 2} SG laser pulse and a LG pulse at a lower peak intensity.« less

  11. Particulate reduction in ternary-compound film growth via pulsed laser deposition from segmented binary-targets

    NASA Astrophysics Data System (ADS)

    Grant-Jacob, James A.; Prentice, Jake J.; Beecher, Stephen J.; Shepherd, David P.; Eason, Robert W.; Mackenzie, Jacob I.

    2018-03-01

    We present the hetero-epitaxial growth of high-quality crystalline Y3Ga5O12 onto a 〈100〉-oriented YAG substrate via pulsed laser deposition, using mixed ternary-compound and segmented binary-compound targets. We observe that a Y3Ga5O12 film fabricated using a segmented target (Y2O3/Ga2O3) contained ∼100 times fewer scattering points than a film grown using a mixed Y3Ga5O12 target. We show that following ablation, the surface of the mixed compound (ternary) target had laser-induced cone structures, whereas the surface of single compound (binary) targets did not. It is concluded that the different ablation dynamics of the oxide constituents in the respective targets plays a significant role in the origin of the scattering points in the resultant films.

  12. Analysis of Current Pulses in HeLa-Cell Permeabilization Due to High Voltage DC Corona Discharge.

    PubMed

    Chetty, Nevendra K; Chonco, Louis; Ijumba, Nelson M; Chetty, Leon; Govender, Thavendran; Parboosing, Raveen; Davidson, Innocent E

    2016-09-01

    Corona discharges are commonly utilized for numerous practical applications, including bio-technological ones. The corona induced transfer of normally impermeant molecules into the interior of biological cells has recently been successfully demonstrated. The exact nature of the interaction of the corona discharge with a cell membrane is still unknown, however, previous studies have suggested that it is either the electric fields produced by ions or the chemical interaction of the reactive species that result in the disruption of the cell membrane. This disruption of the cell membrane allows molecules to permeate into the cell. Corona discharge current constitutes a series of pulses, and it is during these pulses that the ions and reactive species are produced. It stands to reason, therefore, that the nature of these corona pulses would have an influence on the level of cell permeabilization and cell destruction. In this investigation, an analysis of the width, rise-time, characteristic frequencies, magnitude, and repetition rate of the nanosecond pulses was carried out in order to establish the relationship between these factors and the levels of cell membrane permeabilization and cell destruction. Results obtained are presented and discussed.

  13. Influence of Pulsed Electric Fields and Mitochondria-Cytoskeleton Interactions on Cell Respiration.

    PubMed

    Goswami, Ishan; Perry, Justin B; Allen, Mitchell E; Brown, David A; von Spakovsky, Michael R; Verbridge, Scott S

    2018-06-19

    Pulsed electric fields with microsecond pulse width (μsPEFs) are used clinically; namely, irreversible electroporation/Nanoknife is used for soft tissue tumor ablation. The μsPEF pulse parameters used in irreversible electroporation (0.5-1 kV/cm, 80-100 pulses, ∼100 μs each, 1 Hz frequency) may cause an internal field to develop within the cell because of the disruption of the outer cell membrane and subsequent penetration of the electric field. An internal field may disrupt voltage-sensitive mitochondria, although the research literature has been relatively unclear regarding whether such disruptions occur with μsPEFs. This investigation reports the influence of clinically used μsPEF parameters on mitochondrial respiration in live cells. Using a high-throughput Agilent Seahorse machine, it was observed that μsPEF exposure comprising 80 pulses with amplitudes of 600 or 700 V/cm did not alter mitochondrial respiration in 4T1 cells measured after overnight postexposure recovery. To record alterations in mitochondrial function immediately after μsPEF exposure, high-resolution respirometry was used to measure the electron transport chain state via responses to glutamate-malate and ADP and mitochondrial membrane potential via response to carbonyl cyanide-p-trifluoromethoxyphenylhydrazone. In addition to measuring immediate mitochondrial responses to μsPEF exposure, measurements were also made on cells permeabilized using digitonin and those with compromised cytoskeleton due to actin depolymerization via treatment with the drug latrunculin B. The former treatment was used as a control to tease out the effects of plasma membrane permeabilization, whereas the latter was used to investigate indirect effects on the mitochondria that may occur if μsPEFs impact the cytoskeleton on which the mitochondria are anchored. Based on the results, it was concluded that within the pulse parameters tested, μsPEFs alone do not hinder mitochondrial physiology but can be used

  14. Oxygen saturation targeting by pulse oximetry in the extremely low gestational age neonate: a quixotic quest.

    PubMed

    Cummings, James J; Lakshminrusimha, Satyan

    2017-04-01

    A collaboration of comparative effectiveness research trials of pulse oximeter saturation (SpO2) targeting in extremely low-gestational-age neonates have begun to report their aggregate results. We examine the results of those trials, collectively referred to as the Neonatal Oxygenation Prospective Meta-analysis or NeOProM. We also discuss the uncertainties that remain and the clinical challenges that lie ahead. The primary outcome from NeOProM was a composite of death or disability at 18-24 months corrected age. In 2016 the last of these reports was published. Although there were no differences in the primary outcome overall, analyses of secondary outcomes and data subsets following a pulse oximeter revision show significant treatment differences between targeting a lower compared with a higher SpO2. NeOProM represents the largest collaborative clinical research study of SpO2 targets in extremely low-gestational-age neonates. Although aggregate results give us some insight into the feasibility and efficacy of SpO2 targeting in this population, many questions remain. A patient-level analysis, tracking individual outcomes based on actual SpO2 experienced, may shed some light on these questions. However, finding a single optimal SpO2 range seems unlikely.

  15. Measurement of electromagnetic pulses generated during interactions of high power lasers with solid targets

    NASA Astrophysics Data System (ADS)

    De Marco, M.; Krása, J.; Cikhardt, J.; Pfeifer, M.; Krouský, E.; Margarone, D.; Ahmed, H.; Borghesi, M.; Kar, S.; Giuffrida, L.; Vrana, R.; Velyhan, A.; Limpouch, J.; Korn, G.; Weber, S.; Velardi, L.; Delle Side, D.; Nassisi, V.; Ullschmied, J.

    2016-06-01

    A target irradiated with a high power laser pulse, blows off a large amount of charge and as a consequence the target itself becomes a generator of electromagnetic pulses (EMP) owing to high return current flowing to the ground through the target holder. The first measurement of the magnetic field induced by the neutralizing current reaching a value of a few kA was performed with the use of an inductive target probe at the PALS Laser Facility (Cikhardt et al. Rev. Sci. Instrum. 85 (2014) 103507). A full description of EMP generation should contain information on the spatial distribution and temporal variation of the electromagnetic field inside and outside of the interaction chamber. For this reason, we consider the interaction chamber as a resonant cavity in which different modes of EMP oscillate for hundreds of nanoseconds, until the EMP is transmitted outside through the glass windows and EM waves are attenuated. Since the experimental determination of the electromagnetic field distribution is limited by the number of employed antennas, a mapping of the electromagnetic field has to be integrated with numerical simulations. Thus, this work reports on a detailed numerical mapping of the electromagnetic field inside the interaction chamber at the PALS Laser Facility (covering a frequency spectrum from 100 MHz to 3 GHz) using the commercial code COMSOL Multiphysics 5.2. Moreover we carried out a comparison of the EMP generated in the parallelepiped-like interaction chamber used in the Vulcan Petawatt Laser Facility at the Rutherford Appleton Laboratory, against that produced in the spherical interaction chamber of PALS.

  16. Evolution of Gas Cell Targets for Magnetized Liner Inertial Fusion Experiments at the Sandia National Laboratories PECOS Test Facility

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    Paguio, R. R.; Smith, G. E.; Taylor, J. L.

    Z-Beamlet (ZBL) experiments conducted at the PECOS test facility at Sandia National Laboratories (SNL) investigated the nonlinear processes in laser plasma interaction (or laserplasma instabilities LPI) that complicate the deposition of laser energy by enhanced absorption, backscatter, filamentation and beam-spray that can occur in large-scale laser-heated gas cell targets. These targets and experiments were designed to provide better insight into the physics of the laser preheat stage of the Magnetized Liner Inertial Fusion (MagLIF) scheme being tested on the SNL Z-machine. The experiments aim to understand the tradeoffs between laser spot size, laser pulse shape, laser entrance hole (LEH) windowmore » thickness, and fuel density for laser preheat. Gas cell target design evolution and fabrication adaptations to accommodate the evolving experiment and scientific requirements are also described in this paper.« less

  17. Evolution of Gas Cell Targets for Magnetized Liner Inertial Fusion Experiments at the Sandia National Laboratories PECOS Test Facility

    DOE PAGES

    Paguio, R. R.; Smith, G. E.; Taylor, J. L.; ...

    2017-12-04

    Z-Beamlet (ZBL) experiments conducted at the PECOS test facility at Sandia National Laboratories (SNL) investigated the nonlinear processes in laser plasma interaction (or laserplasma instabilities LPI) that complicate the deposition of laser energy by enhanced absorption, backscatter, filamentation and beam-spray that can occur in large-scale laser-heated gas cell targets. These targets and experiments were designed to provide better insight into the physics of the laser preheat stage of the Magnetized Liner Inertial Fusion (MagLIF) scheme being tested on the SNL Z-machine. The experiments aim to understand the tradeoffs between laser spot size, laser pulse shape, laser entrance hole (LEH) windowmore » thickness, and fuel density for laser preheat. Gas cell target design evolution and fabrication adaptations to accommodate the evolving experiment and scientific requirements are also described in this paper.« less

  18. Membrane nanotubes facilitate long-distance interactions between natural killer cells and target cells

    PubMed Central

    Chauveau, Anne; Aucher, Anne; Eissmann, Philipp; Vivier, Eric; Davis, Daniel M.

    2010-01-01

    Membrane nanotubes are membranous tethers that physically link cell bodies over long distances. Here, we present evidence that nanotubes allow human natural killer (NK) cells to interact functionally with target cells over long distances. Nanotubes were formed when NK cells contacted target cells and moved apart. The frequency of nanotube formation was dependent on the number of receptor/ligand interactions and increased on NK cell activation. Most importantly, NK cell nanotubes contained a submicron scale junction where proteins accumulated, including DAP10, the signaling adaptor that associates with the activating receptor NKG2D, and MHC class I chain-related protein A (MICA), a cognate ligand for NKG2D, as occurs at close intercellular synapses between NK cells and target cells. Quantitative live-cell fluorescence imaging suggested that MICA accumulated at small nanotube synapses in sufficient numbers to trigger cell activation. In addition, tyrosine-phosphorylated proteins and Vav-1 accumulated at such junctions. Functionally, nanotubes could aid the lysis of distant target cells either directly or by moving target cells along the nanotube path into close contact for lysis via a conventional immune synapse. Target cells moving along the nanotube path were commonly polarized such that their uropods faced the direction of movement. This is the opposite polarization than for normal cell migration, implying that nanotubes can specifically drive target cell movement. Finally, target cells that remained connected to an NK cell by a nanotube were frequently lysed, whereas removing the nanotube using a micromanipulator reduced lysis of these target cells. PMID:20212116

  19. Investigation into the electromagnetic impulses from long-pulse laser illuminating solid targets inside a laser facility

    NASA Astrophysics Data System (ADS)

    Yi, Tao; Yang, Jinwen; Yang, Ming; Wang, Chuanke; Yang, Weiming; Li, Tingshuai; Liu, Shenye; Jiang, Shaoen; Ding, Yongkun; Xiao, Shaoqiu

    2016-09-01

    Emission of the electromagnetic pulses (EMP) due to laser-target interaction in laser facility had been evaluated using a cone antenna in this work. The microwave in frequencies ranging from several hundreds of MHz to 2 GHz was recorded when long-pulse lasers with several thousands of joules illuminated the solid targets, meanwhile the voltage signals from 1 V to 4 V were captured as functions of laser energy and backlight laser, where the corresponding electric field strengths were obtained by simulating the cone antenna in combination with conducting a mathematical process (Tiknohov Regularization with L curve). All the typical coupled voltage oscillations displayed multiple peaks and had duration of up to 80 ns before decaying into noise and mechanisms of the EMP generation was schematically interpreted in basis of the practical measuring environments. The resultant data were expected to offer basic know-how to achieve inertial confinement fusion.

  20. Cell-specific targeting by heterobivalent ligands.

    PubMed

    Josan, Jatinder S; Handl, Heather L; Sankaranarayanan, Rajesh; Xu, Liping; Lynch, Ronald M; Vagner, Josef; Mash, Eugene A; Hruby, Victor J; Gillies, Robert J

    2011-07-20

    Current cancer therapies exploit either differential metabolism or targeting to specific individual gene products that are overexpressed in aberrant cells. The work described herein proposes an alternative approach--to specifically target combinations of cell-surface receptors using heteromultivalent ligands ("receptor combination approach"). As a proof-of-concept that functionally unrelated receptors can be noncovalently cross-linked with high avidity and specificity, a series of heterobivalent ligands (htBVLs) were constructed from analogues of the melanocortin peptide ligand ([Nle(4), dPhe(7)]-α-MSH) and the cholecystokinin peptide ligand (CCK-8). Binding of these ligands to cells expressing the human Melanocortin-4 receptor and the Cholecystokinin-2 receptor was analyzed. The MSH(7) and CCK(6) were tethered with linkers of varying rigidity and length, constructed from natural and/or synthetic building blocks. Modeling data suggest that a linker length of 20-50 Å is needed to simultaneously bind these two different G-protein coupled receptors (GPCRs). These ligands exhibited up to 24-fold enhancement in binding affinity to cells that expressed both (bivalent binding), compared to cells with only one (monovalent binding) of the cognate receptors. The htBVLs had up to 50-fold higher affinity than that of a monomeric CCK ligand, i.e., Ac-CCK(6)-NH(2). Cell-surface targeting of these two cell types with labeled heteromultivalent ligand demonstrated high avidity and specificity, thereby validating the receptor combination approach. This ability to noncovalently cross-link heterologous receptors and target individual cells using a receptor combination approach opens up new possibilities for specific cell targeting in vivo for therapy or imaging.

  1. Cell-Specific Targeting by Heterobivalent Ligands

    PubMed Central

    Josan, Jatinder S.; Handl, Heather L.; Sankaranarayanan, Rajesh; Xu, Liping; Lynch, Ronald M.; Vagner, Josef; Mash, Eugene A.; Hruby, Victor J.; Gillies, Robert J.

    2012-01-01

    Current cancer therapies exploit either differential metabolism or targeting to specific individual gene products that are overexpressed in aberrant cells. The work described herein proposes an alternative approach—to specifically target combinations of cell-surface receptors using heteromultivalent ligands (“receptor combination approach”). As a proof-of-concept that functionally unrelated receptors can be noncovalently cross-linked with high avidity and specificity, a series of heterobivalent ligands (htBVLs) were constructed from analogues of the melanocortin peptide ligand ([Nle4, DPhe7]-α-MSH) and the cholecystokinin peptide ligand (CCK-8). Binding of these ligands to cells expressing the human Melanocortin-4 receptor and the Cholecystokinin-2 receptor was analyzed. The MSH(7) and CCK(6) were tethered with linkers of varying rigidity and length, constructed from natural and/or synthetic building blocks. Modeling data suggest that a linker length of 20–50 Å is needed to simultaneously bind these two different G-protein coupled receptors (GPCRs). These ligands exhibited up to 24-fold enhancement in binding affinity to cells that expressed both (bivalent binding), compared to cells with only one (monovalent binding) of the cognate receptors. The htBVLs had up to 50-fold higher affinity than that of a monomeric CCK ligand, i.e., Ac-CCK(6)-NH2. Cell-surface targeting of these two cell types with labeled heteromultivalent ligand demonstrated high avidity and specificity, thereby validating the receptor combination approach. This ability to noncovalently cross-link heterologous receptors and target individual cells using a receptor combination approach opens up new possibilities for specific cell targeting in vivo for therapy or imaging. PMID:21639139

  2. Ultra-bright γ-ray flashes and dense attosecond positron bunches from two counter-propagating laser pulses irradiating a micro-wire target.

    PubMed

    Li, Han-Zhen; Yu, Tong-Pu; Hu, Li-Xiang; Yin, Yan; Zou, De-Bin; Liu, Jian-Xun; Wang, Wei-Quan; Hu, Shun; Shao, Fu-Qiu

    2017-09-04

    We propose a novel scheme to generate ultra-bright ultra-short γ-ray flashes and high-energy-density attosecond positron bunches by using multi-dimensional particle-in-cell simulations with quantum electrodynamics effects incorporated. By irradiating a 10 PW laser pulse with an intensity of 10 23 W/cm 2 onto a micro-wire target, surface electrons are dragged-out of the micro-wire and are effectively accelerated to several GeV energies by the laser ponderomotive force, forming relativistic attosecond electron bunches. When these electrons interact with the probe pulse from the other side, ultra-short γ-ray flashes are emitted with an ultra-high peak brightness of 1.8 × 10 24 photons s -1 mm -2 mrad -2 per 0.1%BW at 24 MeV. These photons propagate with a low divergence and collide with the probe pulse, triggering the Breit-Wheeler process. Dense attosecond e - e + pair bunches are produced with the positron energy density as high as 10 17 J/m 3 and number of 10 9 . Such ultra-bright ultra-short γ-ray flashes and secondary positron beams may have potential applications in fundamental physics, high-energy-density physics, applied science and laboratory astrophysics.

  3. Laser imprint reduction for the critical-density foam buffered target driven by a relatively strong foot pulse at early stage of laser implosions

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    Li, J. W., E-mail: li-jiwei@iapcm.ac.cn; He, X. T.; Institute of Applied Physics and Computational Mathematics, P. O. Box 8009, Beijing 100094

    In order to reduce the effect of laser imprint in direct-drive ignition scheme a low-density foam buffered target has been proposed. This target is driven by a laser pulse with a low-intensity foot at the early stage of implosion, which heats the foam and elongates the thermal conduction zone between the laser absorption region and ablation front, increasing the thermal smoothing effect. In this paper, a relatively strong foot pulse is adopted to irradiate the critical-density foam buffered target. The stronger foot, near 1 × 10{sup 14 }W/cm{sup 2}, is able to drive a radiative shock in the low-density foam, which helps smoothmore » the shock and further reduce the effect of laser imprint. The radiative shock also forms a double ablation front structure between the two ablation fronts to further stabilize the hydrodynamics, achieving the similar results to a target with a high-Z dopant in the ablator. 2D analysis shows that for the critical-density foam buffered target irradiated by the strong foot pulse, the laser imprint can be reduced due to the radiative shock in the foam and an increased thermal smoothing effect. It seems viable for the critical-density foam buffered target to be driven by a relatively strong foot pulse with the goal of reducing the laser imprint and achieving better implosion symmetry in the direct-drive laser fusion.« less

  4. Enhancing adoptive cancer immunotherapy with Vγ2Vδ2 T cells through pulse zoledronate stimulation.

    PubMed

    Nada, Mohanad H; Wang, Hong; Workalemahu, Grefachew; Tanaka, Yoshimasa; Morita, Craig T

    2017-01-01

    Human γδ T cells expressing Vγ2Vδ2 T cell receptors monitor foreign- and self-prenyl pyrophosphate metabolites in isoprenoid biosynthesis to mediate immunity to microbes and tumors. Adoptive immunotherapy with Vγ2Vδ2 T cells has been used to treat cancer patients with partial and complete remissions. Most clinical trials and preclinical studies have used continuous zoledronate exposure to expand Vγ2Vδ2 cells where zoledronate is slowly diluted over the course of the culture. Zoledronate inhibits farnesyl diphosphate synthase (FDPS) in monocytes causing isopentenyl pyrophosphate to accumulate that then stimulates Vγ2Vδ2 cells. Because zoledronate inhibition of FDPS is also toxic for T cells, we hypothesized that a short period of exposure would reduce T cell toxicity but still be sufficient for monocytes uptake. Additionally, IL-15 increases the anti-tumor activity of murine αβ T cells in mice but its effect on the in vivo anti-tumor activity of human Vγ2Vδ2 cells has not been assessed. Human Vγ2Vδ2 T cells were expanded by pulse or continuous zoledronate stimulation with IL-2 or IL-15. Expanded Vγ2Vδ2 cells were tested for their expression of effector molecules and killing of tumor cells as well as their in vivo control of human prostate cancer tumors in immunodeficient NSG mice. Pulse zoledronate stimulation with either IL-2 or IL-15 resulted in more uniform expansion of Vγ2Vδ2 cells with higher purity and cell numbers as compared with continuous exposure. The Vγ2Vδ2 cells had higher levels of CD107a and perforin and increased tumor cytotoxicity. Adoptive immunotherapy with Vγ2Vδ2 cells derived by pulse stimulation controlled human PC-3 prostate cancer tumors in NSG mice significantly better than those derived by continuous stimulation, halting tumor growth. Although pulse zoledronate stimulation with IL-15 preserved early memory subsets, adoptive immunotherapy with IL-15-derived Vγ2Vδ2 cells equally inhibited PC-3 tumor growth as those

  5. Slow-Frequency Pulsed Transcranial Electrical Stimulation for Modulation of Cortical Plasticity Based on Reciprocity Targeting with Precision Electrical Head Modeling

    PubMed Central

    Luu, Phan; Essaki Arumugam, Easwara Moorthy; Anderson, Erik; Gunn, Amanda; Rech, Dennis; Turovets, Sergei; Tucker, Don M.

    2016-01-01

    In pain management as well as other clinical applications of neuromodulation, it is important to consider the timing parameters influencing activity-dependent plasticity, including pulsed versus sustained currents, as well as the spatial action of electrical currents as they polarize the complex convolutions of the cortical mantle. These factors are of course related; studying temporal factors is not possible when the spatial resolution of current delivery to the cortex is so uncertain to make it unclear whether excitability is increased or decreased with anodal vs. cathodal current flow. In the present study we attempted to improve the targeting of specific cortical locations by applying current through flexible source-sink configurations of 256 electrodes in a geodesic array. We constructed a precision electric head model for 12 healthy individuals. Extraction of the individual’s cortical surface allowed computation of the component of the induced current that is normal to the target cortical surface. In an effort to replicate the long-term depression (LTD) induced with pulsed protocols in invasive animal research and transcranial magnetic stimulation studies, we applied 100 ms pulses at 1.9 s intervals either in cortical-surface-anodal or cortical-surface-cathodal directions, with a placebo (sham) control. The results showed significant LTD of the motor evoked potential as a result of the cortical-surface-cathodal pulses in contrast to the placebo control, with a smaller but similar LTD effect for anodal pulses. The cathodal LTD after-effect was sustained over 90 min following current injection. These results support the feasibility of pulsed protocols with low total charge in non-invasive neuromodulation when the precision of targeting is improved with a dense electrode array and accurate head modeling. PMID:27531976

  6. Targeting Cell Polarity Machinery to Exhaust Breast Cancer Stem Cells

    DTIC Science & Technology

    2016-10-01

    AWARD NUMBER: W81XWH-15-1-0644 TITLE: Targeting Cell Polarity Machinery to Exhaust Breast Cancer Stem Cells PRINCIPAL INVESTIGATOR: Chun-Ju...U.S. Army Medical Research and Materiel Command Fort Detrick, Maryland 21702-5012 DISTRIBUTION STATEMENT: Approved for Public Release...Targeting Cell Polarity Machinery to Exhaust Breast Cancer Stem Cells 5a. CONTRACT NUMBER 5b. GRANT NUMBER W81XWH-15-1-0644 5c. PROGRAM ELEMENT

  7. Repetitive pulses and laser-induced retinal injury thresholds

    NASA Astrophysics Data System (ADS)

    Lund, David J.

    2007-02-01

    Experimental studies with repetitively pulsed lasers show that the ED 50, expressed as energy per pulse, varies as the inverse fourth power of the number of pulses in the exposure, relatively independently of the wavelength, pulse duration, or pulse repetition frequency of the laser. Models based on a thermal damage mechanism cannot readily explain this result. Menendez et al. proposed a probability-summation model for predicting the threshold for a train of pulses based on the probit statistics for a single pulse. The model assumed that each pulse is an independent trial, unaffected by any other pulse in the train of pulses and assumes that the probability of damage for a single pulse is adequately described by the logistic curve. The requirement that the effect of each pulse in the pulse train be unaffected by the effects of other pulses in the train is a showstopper when the end effect is viewed as a thermal effect with each pulse in the train contributing to the end temperature of the target tissue. There is evidence that the induction of cell death by microcavitation bubbles around melanin granules heated by incident laser irradiation can satisfy the condition of pulse independence as required by the probability summation model. This paper will summarize the experimental data and discuss the relevance of the probability summation model given microcavitation as a damage mechanism.

  8. Direct regulatory immune activity of lactic acid bacteria on Der p 1-pulsed dendritic cells from allergic patients.

    PubMed

    Pochard, Pierre; Hammad, Hamida; Ratajczak, Céline; Charbonnier-Hatzfeld, Anne-Sophie; Just, Nicolas; Tonnel, André-Bernard; Pestel, Joël

    2005-07-01

    Lactic acid bacteria (LAB) are suggested to play a regulatory role in the development of allergic reactions. However, their potential effects on dendritic cells (DCs) directing the immune polarization remain unclear. The immunologic effect of Lactobacillus plantarum NCIMB 8826 (LAB1) on monocyte-derived dendritic cells (MD-DCs) from patients allergic to house dust mite was evaluated. MD-DCs were stimulated for 24 hours with the related allergen Der p 1 in the presence or absence of LAB1. Cell-surface markers were assessed by means of FACS analysis, and the key polarizing cytokines IL-12 and IL-10 were quantified. The subsequent regulatory effect of pulsed MD-DCs on naive or memory T cells was evaluated by determining the T-cell cytokine profile. LAB1 induced the maturation of MD-DCs, even if pulsed with Der p 1. Interestingly, after incubation with LAB1 and Der p 1, MD-DCs produced higher amounts of IL-12 than Der p 1-pulsed DCs. Indeed, the T H 2 cytokine (IL-4 and IL-5) production observed when naive or memory autologous T cells were cocultured with Der p 1-pulsed MD-DCs was highly reduced in the presence of LAB1. Finally, in contrast to naive or memory T cells exposed once to Der p 1-pulsed DCs, T cells stimulated by MD-DCs pulsed with Der p 1 and LAB1 failed to produce T H 2 cytokines in response to a new stimulation with Der p 1-pulsed DCs. Thus in the presence of LAB1, MD-DCs from allergic patients tend to reorientate the T-cell response toward a beneficial T H 1 profile.

  9. A hypothesis of target cell formation in sickle cell disease.

    PubMed

    Wong, P

    2016-08-01

    A fraction of erythrocytes appear as target cells in stained blood smears in sickle cell disease, due to a inheritance of the hemoglobin variant Hb S, polymerizing upon deoxygenation. These cells appear in a three dimension as thin cups. A process of their formation in this disease is proposed based on a band 3-based mechanism of the erythrocyte shape control, able to explain the erythrocyte echinocytosis by glucose depletion. It indicates that their formation is due to a stomatocytogenic slow outward transport of the dibasic form of endogenous Pi with an H(+) by band 3, promoted by the decrease of the Donnan ratio, which decreases cell pH and volume, attributed by a decrease of cell KCl concentration by the higher efflux of K(+)Cl(-) cotransport and Ca(2+) activation of the Gardos channel. Its implications are briefly discussed with respect to target cells per se, target cell formation in other hemoglobinopathies, acquired and inherited disorders of the lipid metabolism and dehydrated hereditary stomatocytosis as well as a stomatocyte presence in a double heterozygote of Hb S and Hb C and of an involvement of the process of target cell formation in acanthocytosis in acquired and inherited disorders. Copyright © 2016 Elsevier Ltd. All rights reserved.

  10. Pulsed source ion implantation apparatus and method

    DOEpatents

    Leung, K.N.

    1996-09-24

    A new pulsed plasma-immersion ion-implantation apparatus that implants ions in large irregularly shaped objects to controllable depth without overheating the target, minimizing voltage breakdown, and using a constant electrical bias applied to the target. Instead of pulsing the voltage applied to the target, the plasma source, for example a tungsten filament or a RF antenna, is pulsed. Both electrically conducting and insulating targets can be implanted. 16 figs.

  11. Pulsed source ion implantation apparatus and method

    DOEpatents

    Leung, Ka-Ngo

    1996-01-01

    A new pulsed plasma-immersion ion-implantation apparatus that implants ions in large irregularly shaped objects to controllable depth without overheating the target, minimizing voltage breakdown, and using a constant electrical bias applied to the target. Instead of pulsing the voltage applied to the target, the plasma source, for example a tungsten filament or a RF antenna, is pulsed. Both electrically conducting and insulating targets can be implanted.

  12. Noncontact microsurgery of cell membranes using femtosecond laser pulses for optoinjection of specified substances into cells

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    Il'ina, I V; Ovchinnikov, A V; Chefonov, O V

    IR femtosecond laser pulses were used for microsurgery of a cell membrane aimed at local and short-duration change in its permeability and injection of specified extracellular substances into the cells. The possibility of noncontact laser delivery of the propidium iodide fluorescent dye and the pEGFP plasmid, encoding the green fluorescent protein, into the cells with preservation of the cell viability was demonstrated. (extreme light fields and their applications)

  13. Evolving phage vectors for cell targeted gene delivery.

    PubMed

    Larocca, David; Burg, Michael A; Jensen-Pergakes, Kristen; Ravey, Edward Prenn; Gonzalez, Ana Maria; Baird, Andrew

    2002-03-01

    We adapted filamentous phage vectors for targeted gene delivery to mammalian cells by inserting a mammalian reporter gene expression cassette (GFP) into the vector backbone and fusing the pIII coat protein to a cell targeting ligand (i.e. FGF2, EGF). Like transfection with animal viral vectors, targeted phage gene delivery is concentration, time, and ligand dependent. Importantly, targeted phage particles are specific for the appropriate target cell surface receptor. Phage have distinct advantages over existing gene therapy vectors because they are simple, economical to produce at high titer, have no intrinsic tropism for mammalian cells, and are relatively simple to genetically modify and evolve. Initially transduction by targeted phage particles was low resulting in foreign gene expression in 1-2% of transfected cells. We increased transduction efficiency by modifying both the transfection protocol and vector design. For example, we stabilized the display of the targeting ligand to create multivalent phagemid-based vectors with transduction efficiencies of up to 45% in certain cell lines when combined with genotoxic treatment. Taken together, these studies establish that the efficiency of phage-mediated gene transfer can be significantly improved through genetic modification. We are currently evolving phage vectors with enhanced cell targeting, increased stability, reduced immunogenicity and other properties suitable for gene therapy.

  14. In vitro stimulation with a strongly pulsed electromagnetic field on rat basophilic leukemia cells

    NASA Astrophysics Data System (ADS)

    Choi, J. W.; Shin, S. C.; Kim, S.; Chung, E. R.; Bang, J. H.; Cho, G. I.; Choi, S. D.; Park, Y. S.; Jang, T. S.; Yoo, Y. M.; Lee, S. S.; Hwang, D. G.

    2010-05-01

    In this study, the effects of pulsed electromagnetic field stimulation with a strong magnetic field on rat basophilic leukemia (RBL-2H3) cells were investigated to confirm the efficacy of the magnetic stimulator for biomedical applications. The maximum intensity of the magnetic field generated from the stimulation coil was 0.203 T, and the transition time was 126 μs. The oscillation time and frequency of the pulsed field were almost 0.1 ms and 8 kHz, respectively. The cell count as well as the mRNA expression and DNA sequence of the cytokine genes, such as the tumor necrosis factor-α (TNF-α) and interleukin-4 (IL-4), of the stimulated RBL-2H3 cells were analyzed with a hemocytometer and via reverse transcriptase polymerase chain reaction to determine the physiological response under a strong pulse field. After 12 h stimulation, cell death was observed at an increasing scale with the increase in the stimulation time. On the other hand, the cells that were stimulated for 10 min almost doubled as the interval time between the stimulations was extended.

  15. P-doped strontium titanate grown using two target pulsed laser deposition for thin film solar cells

    NASA Astrophysics Data System (ADS)

    Man, Hamdi

    Thin-film solar cells made of Mg-doped SrTiO3 p-type absorbers are promising candidates for clean energy generation. This material shows p-type conductivity and also demonstrates reasonable absorption of light. In addition, p-type SrTiO3 can be deposited as thin films so that the cost can be lower than the competing methods. In this work, Mg-doped SrTiO3 (STO) thin-films were synthesized and analyzed in order to observe their potential to be employed as the base semiconductor in photovoltaic applications. Mg-doped STO thin-films were grown by using pulsed laser deposition (PLD) using a frequency quadrupled Yttrium Aluminum Garnet (YAG) laser and with a substrate that was heated by back surface absorption of infrared (IR) laser light. The samples were characterized using X-ray photoelectron spectroscopy (XPS) and it was observed that Mg atoms were doped successfully in the stoichiometry. Reflection high energy electron diffraction (RHEED) spectroscopy proved that the thin films were polycrystalline. Kelvin Probe work function measurements indicated that the work function of the films were 4.167 eV after annealing. UV/Vis Reflection spectroscopy showed that Mg-doped STO thin-films do not reflect significantly except in the ultraviolet region of the spectrum where the reflection percentage increased up to 80%. Self-doped STO thin-films, Indium Tin Oxide (ITO) thin films and stainless steel foil (SSF) were studied in order to observe their characteristics before employing them in Mg-doped STO based solar cells. Self-doped STO thin films were grown using PLD and the results showed that they are capable of serving as the n-type semiconductor in solar cell applications with oxygen vacancies in their structure and low reflectivity. Indium Tin Oxide thin-films grown by PLD system showed low 25-50 ?/square sheet resistance and very low reflection features. Finally, commercially available stainless steel foil substrates were excellent substrates for the inexpensive growth of

  16. Renal Allograft Survival in Nonhuman Primates Infused With Donor Antigen-Pulsed Autologous Regulatory Dendritic Cells.

    PubMed

    Ezzelarab, M B; Raich-Regue, D; Lu, L; Zahorchak, A F; Perez-Gutierrez, A; Humar, A; Wijkstrom, M; Minervini, M; Wiseman, R W; Cooper, D K C; Morelli, A E; Thomson, A W

    2017-06-01

    Systemic administration of autologous regulatory dendritic cells (DCreg; unpulsed or pulsed with donor antigen [Ag]), prolongs allograft survival and promotes transplant tolerance in rodents. Here, we demonstrate that nonhuman primate (NHP) monocyte-derived DCreg preloaded with cell membrane vesicles from allogeneic peripheral blood mononuclear cells induce T cell hyporesponsiveness to donor alloantigen (alloAg) in vitro. These donor alloAg-pulsed autologous DCreg (1.4-3.6 × 10 6 /kg) were administered intravenously, 1 day before MHC-mismatched renal transplantation to rhesus monkeys treated with costimulation blockade (cytotoxic T lymphocyte Ag 4 immunoglobulin [CTLA4] Ig) and tapered rapamycin. Prolongation of graft median survival time from 39.5 days (no DCreg infusion; n = 6 historical controls) and 29 days with control unpulsed DCreg (n = 2), to 56 days with donor Ag-pulsed DCreg (n = 5) was associated with evidence of modulated host CD4 + and CD8 + T cell responses to donor Ag and attenuation of systemic IL-17 production. Circulating anti-donor antibody (Ab) was not detected until CTLA4 Ig withdrawal. One monkey treated with donor Ag-pulsed DCreg rejected its graft in association with progressively elevated anti-donor Ab, 525 days posttransplant (160 days after withdrawal of immunosuppression). These findings indicate a modest but not statistically significant beneficial effect of donor Ag-pulsed autologous DCreg infusion on NHP graft survival when administered with a minimal immunosuppressive drug regimen. © 2016 The American Society of Transplantation and the American Society of Transplant Surgeons.

  17. Preliminary ex vivo feasibility study on targeted cell surgery by high intensity focused ultrasound (HIFU).

    PubMed

    Wang, Zhi Biao; Wu, Junru; Fang, Liao Qiong; Wang, Hua; Li, Fa Qi; Tian, Yun Bo; Gong, Xiao Bo; Zhang, Hong; Zhang, Lian; Feng, Ruo

    2011-04-01

    High intensity focused ultrasound (HIFU) has become a new noninvasive surgical modality in medicine. A portion of tissue seated inside a patient's body may experience coagulative necrosis after a few seconds of insonification by high intensity focused ultrasound (US) generated by an extracorporeal focusing US transducer. The region of tissue affected by coagulative necrosis (CN) usually has an ellipsoidal shape when the thermal effect due to US absorption plays the dominant role. Its long and short axes are parallel and perpendicular to the US propagation direction respectively. It was shown by numerical computations using a nonlinear Gaussian beams model to describe the sound field in a focal zone and ex vivo experiments that the dimension of the short and long axes of the tissue which experiences CN can be as small as 50μm and 250μm respectively after one second exposure of US pulse (the spatial and pulse average acoustic power is on the order of tens of Watts and the local acoustic spatial and temporal pulse averaged intensity is on the order of 3×10(4)W/cm(2)) generated by a 1.6MHz HIFU transducer of 12cm diameter and 11cm geometric focal length (f-number=0.92). The concept of thermal dose of cumulative equivalent minutes was used to describe the possible tissue coagulative necrosis generated by HIFU. The numbers of cells which suffered CN were estimated to be on the order of 40. This result suggests that HIFU is able to interact with tens of cells at/near its focal zone while keeping the neighboring cells minimally affected, and thus the targeted cell surgery may be achievable. Copyright © 2010 Elsevier B.V. All rights reserved.

  18. Suppression of suprathermal ions from a colloidal microjet target containing SnO2 nanoparticles by using double laser pulses

    NASA Astrophysics Data System (ADS)

    Higashiguchi, Takeshi; Kaku, Masanori; Katto, Masahito; Kubodera, Shoichi

    2007-10-01

    We have demonstrated suppression of suprathermal ions from a colloidal microjet target plasma containing tin-dioxide (SnO2) nanoparticles irradiated by double laser pulses. We observed a significant decrease of the tin and oxygen ion signals in the charged-state-separated energy spectra when double laser pulses were irradiated. The peak energy of the singly ionized tin ions decreased from 9to3keV when a preplasma was produced. The decrease in the ion energy, considered as debris suppression, is attributed to the interaction between an expanding low-density preplasma and a main laser pulse.

  19. Pulsed laser activated cell sorter (PLACS) for high-throughput fluorescent mammalian cell sorting

    NASA Astrophysics Data System (ADS)

    Chen, Yue; Wu, Ting-Hsiang; Chung, Aram; Kung, Yu-Chung; Teitell, Michael A.; Di Carlo, Dino; Chiou, Pei-Yu

    2014-09-01

    We present a Pulsed Laser Activated Cell Sorter (PLACS) realized by exciting laser induced cavitation bubbles in a PDMS microfluidic channel to create high speed liquid jets to deflect detected fluorescent samples for high speed sorting. Pulse laser triggered cavitation bubbles can expand in few microseconds and provide a pressure higher than tens of MPa for fluid perturbation near the focused spot. This ultrafast switching mechanism has a complete on-off cycle less than 20 μsec. Two approaches have been utilized to achieve 3D sample focusing in PLACS. One is relying on multilayer PDMS channels to provide 3D hydrodynamic sheath flows. It offers accurate timing control of fast (2 m sec-1) passing particles so that synchronization with laser bubble excitation is possible, an critically important factor for high purity and high throughput sorting. PLACS with 3D hydrodynamic focusing is capable of sorting at 11,000 cells/sec with >95% purity, and 45,000 cells/sec with 45% purity using a single channel in a single step. We have also demonstrated 3D focusing using inertial flows in PLACS. This sheathless focusing approach requires 10 times lower initial cell concentration than that in sheath-based focusing and avoids severe sample dilution from high volume sheath flows. Inertia PLACS is capable of sorting at 10,000 particles sec-1 with >90% sort purity.

  20. Statistical Modeling of Single Target Cell Encapsulation

    PubMed Central

    Moon, SangJun; Ceyhan, Elvan; Gurkan, Umut Atakan; Demirci, Utkan

    2011-01-01

    High throughput drop-on-demand systems for separation and encapsulation of individual target cells from heterogeneous mixtures of multiple cell types is an emerging method in biotechnology that has broad applications in tissue engineering and regenerative medicine, genomics, and cryobiology. However, cell encapsulation in droplets is a random process that is hard to control. Statistical models can provide an understanding of the underlying processes and estimation of the relevant parameters, and enable reliable and repeatable control over the encapsulation of cells in droplets during the isolation process with high confidence level. We have modeled and experimentally verified a microdroplet-based cell encapsulation process for various combinations of cell loading and target cell concentrations. Here, we explain theoretically and validate experimentally a model to isolate and pattern single target cells from heterogeneous mixtures without using complex peripheral systems. PMID:21814548

  1. Targeted femtosecond laser driven drug delivery within HIV-1 infected cells: in-vitro studies

    NASA Astrophysics Data System (ADS)

    Maphanga, Charles; Ombinda-Lemboumba, Saturnin; Manoto, Sello; Maaza, Malik; Mthunzi-Kufa, Patience

    2017-02-01

    Human immunodeficiency virus (HIV-1) infection still remains one amongst the world's most challenging infections since its discovery. Antiretroviral therapy is the recommended treatment of choice for HIV-1 infection taken by patients orally. The highly active antiretroviral therapy (HAART) prevents the replication of HIV-1 and further destruction of the immune system, therefore enabling the body to fight opportunistic life-threatening infections, cancers, and also arrest HIV infection from advancing to AIDS. The major challenge with HAART is the inability to reach the viral reservoirs where the HIV-1 remains latent and persistent, leading to inability to fully eradicate the virus. This study is aimed at initially designing and assembling a fully functional optical translocation setup to optically deliver antiretroviral drugs into HIV-1 infected cells in a targeted manner using Gaussian beam mode femtosecond laser pulses in-vitro. The main objective of our study is to define the in-vitro drug photo-translocation parameters to allow future design of an efficient drug delivery device with potential in-vivo drug delivery applications. In our experiments, HEK 293T cells were used to produce HIV-1 enveloped pseudovirus (ZM53) to infect TZM-bl cells which were later treated with laser pulses emitted by a titanium sapphire laser (800 nm, 1KHz, 113 fs, 6.5 μW) to create sub-microscopic pores on the cell membrane enabling influx of extracellular media. Following laser treatment, changes in cellular responses were analysed using cell morphology studies, cytotoxicity, and luciferase assay studies. Controls included laser untreated cells incubated with the drug for 72 hours. The data in this study was statistically analysed using the SigmaPlot software version 13.

  2. Effects of nanosecond pulsed electrical fields (nsPEFs) on the cell cycle of CHO and Jurkat cells

    NASA Astrophysics Data System (ADS)

    Mahlke, Megan A.; Navara, Christopher; Ibey, Bennett L.

    2014-03-01

    Exposure to nano-second pulsed electrical fields (nsPEFs) can cause poration of external and internal cell membranes, DNA damage, and disassociation of cytoskeletal components, all of which are capable of disrupting a cell's ability to replicate. Variations between cell lines in membrane and cytoskeletal structure as well as in survival of nsPEF exposure should correspond to unique line-dependent cell cycle effects. Additionally, phase of cell cycle during exposure may be linked to differential sensitivities to nsPEFs across cell lines, as DNA structure, membrane elasticity, and cytoskeletal structure change dramatically during the cell cycle. Populations of Jurkat and Chinese Hamster Ovary (CHO) cells were examined post-exposure (10 ns pulse trains at 150kV/cm) by analysis of DNA content via propidium iodide staining and flow cytometric analysis at various time points (1, 6, and 12h post-exposure) to determine population distribution in cell cycle phases. Additionally, CHO and Jurkat cells were synchronized in G1/S and G2/M phases, pulsed, and analyzed to evaluate role of cell cycle phase in survival of nsPEFs. CHO populations recovered similarly to sham populations postnsPEF exposure and did not exhibit a phase-specific change in response. Jurkat cells exhibited considerable apoptosis/necrosis in response to nsPEF exposure and were unable to recover and proliferate in a manner similar to sham exposed cells. Additionally, Jurkat cells appear to be more sensitive to nsPEFs in G2/M phases than in G1/S phases. Recovery of CHO populations suggests that nsPEFs do not inhibit proliferation in CHO cells; however, inhibition of Jurkat cells post-nsPEF exposure coupled with preferential cell death in G2/M phases suggest that cell cycle phase during exposure may be an important factor in determining nsPEF toxicity in certain cell lines. Interestingly, CHO cells have a more robust and rigid cytoskeleton than Jurkat cells which is thought to contribute to their ability to

  3. Oxidant-induced DNA damage of target cells.

    PubMed Central

    Schraufstätter, I; Hyslop, P A; Jackson, J H; Cochrane, C G

    1988-01-01

    In this study we examined the leukocytic oxidant species that induce oxidant damage of DNA in whole cells. H2O2 added extracellularly in micromolar concentrations (10-100 microM) induced DNA strand breaks in various target cells. The sensitivity of a specific target cell was inversely correlated to its catalase content and the rate of removal of H2O2 by the target cell. Oxidant species produced by xanthine oxidase/purine or phorbol myristate acetate-stimulated monocytes induced DNA breakage of target cells in proportion to the amount of H2O2 generated. These DNA strand breaks were prevented by extracellular catalase, but not by superoxide dismutase. Cytotoxic doses of HOCl, added to target cells, did not induce DNA strand breakage, and myeloperoxidase added extracellularly in the presence of an H2O2-generating system, prevented the formation of DNA strand breaks in proportion to its H2O2 degrading capacity. The studies also indicated that H2O2 formed hydroxyl radical (.OH) intracellularly, which appeared to be the most likely free radical responsible for DNA damage: .OH was detected in cells exposed to H2O2; the DNA base, deoxyguanosine, was hydroxylated in cells exposed to H2O2; and intracellular iron was essential for induction of DNA strand breaks. PMID:2843565

  4. A hormone pulse induces transient changes in the subcellular distribution and leads to a lysosomal accumulation of the estradiol receptor alpha in target tissues.

    PubMed

    Qualmann, B; Kessels, M M; Thole, H H; Sierralta, W D

    2000-06-01

    An intrauterine pulse-stimulation with estradiol induced changes in the subcellular localization of estrogen receptor alpha in porcine endometrium, as detected with F(ab') fragments of various anti-receptor antibodies covalently linked to nanogold. The low-sterically hindered immunoreagents--recognizing different epitopes within the hormone binding domain--allowed for an efficient immunolabeling of estradiol receptor alpha, detecting it both in the cytoplasm and the nucleus of nonstimulated epithelium cells. In the cytoplasm, the receptor often seemed to be associated with actin filaments and the endoplasmatic reticulum. After the stimulation with estradiol, a predominantly nuclear localization and a labeling of nucleoli was observed. Our immunoelectron microscopy study demonstrates a localization of the receptor in cytoplasmic organelles that increased after the hormone pulse. These organelles exhibited the morphological properties of lysosomes and relocated to the perinuclear area. In analogous cytoplasmic organelles, the presence of cathepsin D was detected via indirect immunogold labeling, justifying their classification as lysosomes. Quantitative examinations revealed that not only the number of lysosomes in the proximity of the nucleus but also their immunostaining for estradiol receptor alpha increased significantly after the hormone pulse. Thus, estradiol induces both the rapid shift of receptor into the nucleus, a slower perinuclear accumulation of lysosomes and an increase of lysosomal ERalpha-immunoreactivity. These results suggest a role for lysosomes in the degradation of receptor shuttling out of the nucleus. This could serve as termination of the estradiol receptor alpha-dependent activation of target cells. This hypothesis is strengthened by the fact that the receptor content in uterine tissue declined drastically few hours after the hormone pulse.

  5. Renal Allograft Survival in Nonhuman Primates Infused with Donor Antigen-Pulsed Autologous Regulatory Dendritic Cells

    PubMed Central

    Ezzelarab, M.B.; Raich-Regue, D.; Lu, L.; Zahorchak, A.F.; Perez-Gutierrez, A.; Humar, A.; Wijkstrom, M.; Minervini, M.; Wiseman, R.W.; Cooper, D.K.C.; Morelli, A.E.; Thomson, A.W.

    2017-01-01

    Systemic administration of autologous regulatory dendritic cells (DCreg; unpulsed or pulsed with donor antigen [Ag]), prolongs allograft survival and promotes transplant tolerance in rodents. Here, we demonstrate that nonhuman primate (NHP) monocyte-derived DCreg pre-loaded with cell membrane vesicles from allogeneic PBMC, induce T cell hyporesponsiveness to donor alloAg in vitro. These donor alloAg-pulsed autologous DCreg (1.4–3.6 x 106/kg) were administered intravenously, one day before MHC-mismatched renal transplantation to rhesus monkeys treated with costimulation blockade (cytotoxic T lymphocyte Ag 4 [CTLA4] Ig) and tapered rapamycin. Prolongation of graft median survival time from 39.5 days (no DCreg infusion; n=6 historical controls) and 29 days with control unpulsed DCreg (n=2), to 56 days with donor Ag-pulsed DCreg (n=5), was associated with evidence of modulated host CD4+ and CD8+ T cell responses to donor Ag and attenuation of systemic IL-17 production. Circulating anti-donor antibody (Ab) was not detected until CTLA4Ig withdrawal. One monkey treated with donor Ag-pulsed DCreg rejected its graft in association with progressively elevated anti-donor Ab, 525 days post-transplant (160 days after withdrawal of immunosuppression). These findings indicate a modest but not statistically significant beneficial effect of donor Ag-pulsed autologous DCreg infusion on NHP graft survival when administered with a minimal immunosuppressive drug regimen. PMID:28009481

  6. Laser lift-off scribing of the CZTSe thin-film solar cells at different pulse durations

    DOE PAGES

    Markauskas, Edgaras; Gečys, Paulius; Repins, Ingrid; ...

    2017-04-27

    Here, the transition to fully sized solar modules requires additional three-step laser structuring processes to preserve small-scale cell efficiencies over the large areas. The adjacent cell isolation (the P3 scribe) was found to be the most sensitive process in the case of laser induced damage. The laser induced layer lift-off mechanism seems to be a very attractive process for the P3 patterning, since almost all the laser affected material is removed by mechanical spallation. However, a laser induced layer spallation behavior together with scribe electrical validation under the different laser pulse durations was not investigated extensively in the past. Therefore,more » we report our novel results on the P2 and P3 laser lift-off processing of the Cu 2ZnSn(S, Se 4) (CZTSe) thin-film solar cells covering the pulse duration range from 300 fs to 60 ps. Shorter sub-ps pulses enabled us to process smaller P2 and P3 craters, although the lift-off threshold fluences were higher compared to the longer ps pulses. In the case of the layer lift-off, the laser radiation had to penetrate through the layer stack down to the CZTSe/Mo interface. At shorter sub-ps pulses, the nonlinear effects triggered absorption of the laser radiation in the bulk of the material, resulting in increased damage of the CZTSe layer. The Raman measurements confirmed the CZTSe surface stoichiometry changes for shorter pulses. Furthermore, shorter pulses induced higher electrical conductivity of a scribe, resulting in lower photo-electrical efficiency during the mini-module simulation. In the case of the P3 lift-off scribing, the 10 ps pulses were more favorable than shorter femtosecond pulses.« less

  7. Laser lift-off scribing of the CZTSe thin-film solar cells at different pulse durations

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    Markauskas, Edgaras; Gečys, Paulius; Repins, Ingrid

    Here, the transition to fully sized solar modules requires additional three-step laser structuring processes to preserve small-scale cell efficiencies over the large areas. The adjacent cell isolation (the P3 scribe) was found to be the most sensitive process in the case of laser induced damage. The laser induced layer lift-off mechanism seems to be a very attractive process for the P3 patterning, since almost all the laser affected material is removed by mechanical spallation. However, a laser induced layer spallation behavior together with scribe electrical validation under the different laser pulse durations was not investigated extensively in the past. Therefore,more » we report our novel results on the P2 and P3 laser lift-off processing of the Cu 2ZnSn(S, Se 4) (CZTSe) thin-film solar cells covering the pulse duration range from 300 fs to 60 ps. Shorter sub-ps pulses enabled us to process smaller P2 and P3 craters, although the lift-off threshold fluences were higher compared to the longer ps pulses. In the case of the layer lift-off, the laser radiation had to penetrate through the layer stack down to the CZTSe/Mo interface. At shorter sub-ps pulses, the nonlinear effects triggered absorption of the laser radiation in the bulk of the material, resulting in increased damage of the CZTSe layer. The Raman measurements confirmed the CZTSe surface stoichiometry changes for shorter pulses. Furthermore, shorter pulses induced higher electrical conductivity of a scribe, resulting in lower photo-electrical efficiency during the mini-module simulation. In the case of the P3 lift-off scribing, the 10 ps pulses were more favorable than shorter femtosecond pulses.« less

  8. Nanosurgery with near-infrared 12-femtosecond and picosecond laser pulses

    NASA Astrophysics Data System (ADS)

    Uchugonova, Aisada; Zhang, Huijing; Lemke, Cornelius; König, Karsten

    2011-03-01

    Laser-assisted surgery based on multiphoton absorption of NIR laser light has great potential for high precision surgery at various depths within the cells and tissues. Clinical applications include refractive surgery (fs-LASIK). The non-contact laser method also supports contamination-free cell nanosurgery. Here we apply femtosecond laser scanning microscopes for sub-100 nm surgery of human cells and metaphase chromosomes. A mode-locked 85 MHz Ti:Sapphire laser with an M-shaped ultrabroad band spectrum (maxima: 770 nm/830 nm) with an in situ pulse duration at the target ranging from 12 femtoseconds up to 3 picoseconds was employed. The effects of laser nanoprocessing in cells and chromosomes have been quantified by atomic force microscopy (AFM) and electron microscopy. These studies demonstrate the potential of extreme ultrashort femtosecond laser pulses at low mean milliwatt powers for sub-100 nm surgery.

  9. Optical cell cleaning with NIR femtosecond laser pulses

    NASA Astrophysics Data System (ADS)

    Uchugonova, Aisada; Breunig, Hans Georg; Batista, Ana; König, Karsten

    2015-03-01

    Femtosecond laser microscopes have been used as both micro and nanosurgery tools. The optical knock-out of undesired cells in multiplex cell clusters shall be further reported on in this study. Femtosecond laser-induced cell death is beneficial due to the reduced collateral side effects and therefore can be used to selectively destroy target cells within monolayers, as well as within 3D tissues, all the while preserving cells of interest. This is an important characteristic for the application in stem cell research and cancer treatment. Non-precise damage compromises the viability of neighboring cells by inducing side effects such as stress to the cells surrounding the target due to the changes in the microenvironment, resulting from both the laser and laser-exposed cells. In this study, optimum laser parameters for optical cleaning by isolating single cells and cell colonies are exploited through the use of automated software control. Physiological equilibrium and cellular responses to the laser induced damages are also investigated. Cell death dependence on laser focus, determination and selectivity of intensity/dosage, controllable damage and cell recovery mechanisms are discussed.

  10. Nanosecond pulsed electric field thresholds for nanopore formation in neural cells

    NASA Astrophysics Data System (ADS)

    Roth, Caleb C.; Tolstykh, Gleb P.; Payne, Jason A.; Kuipers, Marjorie A.; Thompson, Gary L.; DeSilva, Mauris N.; Ibey, Bennett L.

    2013-03-01

    The persistent influx of ions through nanopores created upon cellular exposure to nanosecond pulse electric fields (nsPEF) could be used to modulate neuronal function. One ion, calcium (Ca), is important to action potential firing and regulates many ion channels. However, uncontrolled hyper-excitability of neurons leads to Ca overload and neurodegeneration. Thus, to prevent unintended consequences of nsPEF-induced neural stimulation, knowledge of optimum exposure parameters is required. We determined the relationship between nsPEF exposure parameters (pulse width and amplitude) and nanopore formation in two cell types: rodent neuroblastoma (NG108) and mouse primary hippocampal neurons (PHN). We identified thresholds for nanoporation using Annexin V and FM1-43, to detect changes in membrane asymmetry, and through Ca influx using Calcium Green. The ED50 for a single 600 ns pulse, necessary to cause uptake of extracellular Ca, was 1.76 kV/cm for NG108 and 0.84 kV/cm for PHN. At 16.2 kV/cm, the ED50 for pulse width was 95 ns for both cell lines. Cadmium, a nonspecific Ca channel blocker, failed to prevent Ca uptake suggesting that observed influx is likely due to nanoporation. These data demonstrate that moderate amplitude single nsPEF exposures result in rapid Ca influx that may be capable of controllably modulating neurological function.

  11. Inorganic nanocomposite films with polymer nanofillers made by the concurrent multi-beam multi-target pulsed laser deposition

    NASA Astrophysics Data System (ADS)

    Darwish, Abdalla M.; Sarkisov, Sergey S.; Mele, Paolo; Saini, Shrikant; Moore, Shaelynn; Bastian, Tyler; Dorlus, Wydglif; Zhang, Xiaodong; Koplitz, Brent

    2017-08-01

    We report on the new class of inorganic nanocomposite films with the inorganic phase hosting the polymer nanofillers made by the concurrent multi-beam multi-target pulsed laser deposition of the inorganic target material and matrix assisted pulsed laser evaporation of the polymer (MBMT-PLD/MAPLE). We used the exemplary nanocomposite thermoelectric films of aluminum-doped ZnO known as AZO with the nanofillers made of poly(methyl methacrylate) known as PMMA on various substrates such as SrTiO3, sapphire, fused silica, and polyimide. The AZO target was ablated with the second harmonic (532 nm) of the Nd:YAG Q-switched laser while PMMA was evaporated from its solution in chlorobenzene frozen in liquid nitrogen with the fundamental harmonic (1064 nm) of the same laser (50 Hz pulse repetition rate). The introduction of the polymer nanofillers increased the electrical conductivity of the nanocomposite films (possibly due to the carbonization of PMMA and the creation of additional channels of electric current) three times and reduced the thermal conductivity by 1.25 times as compared to the pure AZO films. Accordingly, the increase of the thermoelectric figure-of merit ZT would be 4 times. The best performance was observed for the sapphire substrates where the films were the most uniform. The results point to a huge potential of the optimization of a broad variety of optical, opto-electronic, and solar-power nanocomposite inorganic films by the controllable introduction of the polymer nanofillers using the MBMT-PLD/MAPLE method.

  12. Efficient neutron production from sub-nanosecond laser pulse accelerating deuterons on target front side

    NASA Astrophysics Data System (ADS)

    Klir, D.; Krasa, J.; Cikhardt, J.; Dudzak, R.; Krousky, E.; Pfeifer, M.; Rezac, K.; Sila, O.; Skala, J.; Ullschmied, J.; Velyhan, A.

    2015-09-01

    Neutron-producing experiments have been carried out on the Prague Asterix Laser System. At the fundamental wavelength of 1.315 μm, the laser pulse of a 600 J energy and 300 ps duration was focused on a thick deuterated-polyethylene target. Neutron yields reached (4.1 ± 0.8) × 108 at the peak intensity of ≈3 × 1016 W/cm2. A more detailed analysis of neutron time-of-flight signals showed that a significant fraction of neutron yields was produced both by the 2H(d,n)3He reaction and by other neutron-producing reactions. Neutron energies together with delayed neutron and gamma emission showed that MeV deuterons escaped from a laser-produced plasma and interacted ≈50 ns later with a borosilicate blast-shield glass. In order to increase DD neutron yields and to characterize deuteron beams via nuclear reactions, a secondary deuterated polyethylene target was used in a pitcher-catcher scheme at the target front side. In this experimental arrangement, the neutron yield reached (2.0 ± 0.5) × 109 with the peak neutron fluence of (2.5 ± 0.5) × 108 n/sr. From the neutron yield, it was calculated that the secondary target was bombarded by 2 × 1014 deuterons in the 0.5-2.0 MeV energy range. The neutron yield of 2 × 109 at the laser energy of 600 J implied the production efficiency of 3 × 106 n/J. A very important result is that the efficient neutron production was achieved with the low contrast, sub-nanosecond laser pulse of the intensity of 1016 W/cm2. The latter parameters can be achieved in a rep-rate mode more easily than ultra-high intensities and contrasts.

  13. Effect of Shock Waves Generated by Pulsed Electric Discharges in Water on Yeast Cells and Virus Particles

    NASA Astrophysics Data System (ADS)

    Girdyuk, A. E.; Gorshkov, A. N.; Egorov, V. V.; Kolikov, V. A.; Snetov, V. N.; Shneerson, G. A.

    2018-02-01

    The aim of this study is to determine the optimal parameters of the electric pulses and shock waves generated by them for the soft destruction of the virus and yeast envelopes with no changes in the structure of antigenic surface albumin and in the cell morphology in order to use them to produce antivirus vaccines and in biotechnology. The pulse electric discharges in water have been studied for different values of amplitude, pulse duration and the rate of the rise in the current. A mathematical model has been developed to estimate the optimal parameters of pulsed electric charges and shock waves for the complete destruction of the yeast cell envelopes and virus particles at a minimum of pulses.

  14. Designing oral vaccines targeting intestinal dendritic cells.

    PubMed

    Devriendt, Bert; De Geest, Bruno G; Cox, Eric

    2011-04-01

    Most pathogens colonize and invade the host at mucosal surfaces, such as the lung and the intestine. To combat intestinal pathogens the induction of local adaptive immune responses is required, which is mainly achieved through oral vaccination. However, most vaccines are ineffective when given orally owing to the hostile environment in the gastrointestinal tract. The encapsulation of antigens in biodegradable microparticulate delivery systems enhances their immunogenicity; however, the uptake of these delivery systems by intestinal immune cells is rather poor. Surface decoration of the particulates with targeting ligands could increase the uptake and mediate the selective targeting of the vaccine to intestinal antigen-presenting cells, including dendritic cells. In this review, current knowledge on dendritic cell subsets is discussed, along with progress in the development of selective antigen targeting to these cells, in addition to focusing on data obtained in mice and, where possible, the pig, as a non-rodent animal model for humans. Moreover, the potential use and benefits of Fcγ receptor-mediated targeting of antigen delivery systems are highlighted. In conclusion, dendritic cell targeting ligands grafted on antigen carrier systems should preferably bind to a conserved endocytotic receptor, facilitating the design of a multispecies vaccine platform, which could elicit robust protective immune responses against enteric pathogens.

  15. Shared target antigens on cancer cells and tissue stem cells: go or no-go for CAR T cells?

    PubMed

    Hombach, Andreas A; Abken, Hinrich

    2017-02-01

    Adoptive therapy with chimeric antigen receptor (CAR) T cells redirected towards CD19 produces remissions of B cell malignancies, however, it also eradicates healthy B cells sharing the target antigen. Such 'on-target off-tumor' toxicity raises serious safety concerns when the target antigen is also expressed by tissue stem cells, with the risk of lasting tissue destruction. Areas covered: We discuss CAR T cell targeting of activation antigens versus lineage associated antigens on the basis of recent experimental and animal data and the literature in the field. Expert commentary: Targeting an activation associated antigen which is transiently expressed by stem cells seems to be safe, like CAR T cells targeting CD30 spare CD30 + hematopoietic stem and progenitor cells while eliminating CD30 + lymphoma cells, whereas targeting lineage associated antigens which increase in expression during cell maturation, like folate receptor-β and CD123, is of risk to destruct tissue stem cells.

  16. Influence of the laser pulse repetition rate and scanning speed on the morphology of Ag nanostructures fabricated by pulsed laser ablation of solid target in water

    NASA Astrophysics Data System (ADS)

    Nikolov, A. S.; Balchev, I. I.; Nedyalkov, N. N.; Kostadinov, I. K.; Karashanova, D. B.; Atanasova, G. B.

    2017-11-01

    Nanostructures of noble metal were produced by pulsed laser ablation in liquid. A solid Ag target was immersed in double distilled water and a CuBr laser in a master oscillator—power amplifier configuration oscillating at 511 nm and emitting pulses with duration of 30 ns at a repetition rate of up to 20 kHz was employed to produce different colloids. The impact was studied of the laser pulse repetition rate and the beam scanning speed on the morphology of the nanostructures formed. Further, the optical extinction spectra of the colloids in the UV/VIS range were measured and used to make an indirect assessment of the changes in the shape and size distribution of the nanostructures. The transmission values in the near UV range were used to estimate the efficiency of the ablation process under the different experimental conditions implemented. A visualization of the nanostructures was made possible by transmission electron microscopy (TEM). The structure and phase composition of the nanoparticles were studied by high-resolution transmission electron microscopy (HRTEM) and selected area electron diffraction (SAED), while the alteration of the target surface caused by the impact of the high-repetition-rate laser illumination was investigated by X-ray photoelectron spectroscopy (XPS). The optimal conditions were determined yielding the highest efficiency in terms of amount of ablated material.

  17. Effects of the pulse width on the reactive species production and DNA damage in cancer cells exposed to atmospheric pressure microsecond-pulsed helium plasma jets

    NASA Astrophysics Data System (ADS)

    Joh, Hea Min; Choi, Ji Ye; Kim, Sun Ja; Kang, Tae Hong; Chung, T. H.

    2017-08-01

    Plasma-liquid and plasma-cell interactions were investigated using an atmospheric pressure dc microsecond-pulsed helium plasma jet. We investigated the effects of the electrical parameters such as applied voltage and pulse width (determined by the pulse frequency and duty ratio) on the production of reactive species in the gas/liquid phases and on the DNA damage responses in the cancer cells. The densities of reactive species including OH radicals were estimated inside the plasma-treated liquids using a chemical probe method, and the nitrite concentration was detected by Griess assay. Importantly, the more concentration of OH resulted in the more DNA base oxidation and breaks in human lung cancer A549 cells. The data are very suggestive that there is strong correlation between the production of OH in the plasmas/liquids and the DNA damage.

  18. Deposition and characterization of titania-silica optical multilayers by asymmetric bipolar pulsed dc sputtering of oxide targets

    NASA Astrophysics Data System (ADS)

    Sagdeo, P. R.; Shinde, D. D.; Misal, J. S.; Kamble, N. M.; Tokas, R. B.; Biswas, A.; Poswal, A. K.; Thakur, S.; Bhattacharyya, D.; Sahoo, N. K.; Sabharwal, S. C.

    2010-02-01

    Titania-silica (TiO2/SiO2) optical multilayer structures have been conventionally deposited by reactive sputtering of metallic targets. In order to overcome the problems of arcing, target poisoning and low deposition rates encountered there, the application of oxide targets was investigated in this work with asymmetric bipolar pulsed dc magnetron sputtering. In order to evaluate the usefulness of this deposition methodology, an electric field optimized Fabry Perot mirror for He-Cd laser (λ = 441.6 nm) spectroscopy was deposited and characterized. For comparison, this mirror was also deposited by the reactive electron beam (EB) evaporation technique. The mirrors developed by the two complementary techniques were investigated for their microstructural and optical reflection properties invoking atomic force microscopy, ellipsometry, grazing incidence reflectometry and spectrophotometry. From these measurements the layer geometry, optical constants, mass density, topography, surface and interface roughness and disorder parameters were evaluated. The microstructural properties and spectral functional characteristics of the pulsed dc sputtered multilayer mirror were found to be distinctively superior to the EB deposited mirror. The knowledge gathered during this study has been utilized to develop a 21-layer high-pass edge filter for radio photoluminescence dosimetry.

  19. Targeting tumor cell motility to prevent metastasis

    PubMed Central

    Palmer, Trenis D.; Ashby, William J.; Lewis, John D.; Zijlstra, Andries

    2011-01-01

    Mortality and morbidity in patients with solid tumors invariably results from the disruption of normal biological function caused by disseminating tumor cells. Tumor cell migration is under intense investigation as the underlying cause of cancer metastasis. The need for tumor cell motility in the progression of metastasis has been established experimentally and is supported empirically by basic and clinical research implicating a large collection of migration-related genes. However, there are few clinical interventions designed to specifically target the motility of tumor cells and adjuvant therapy to specifically prevent cancer cell dissemination is severely limited. In an attempt to define motility targets suitable for treating metastasis, we have parsed the molecular determinants of tumor cell motility into five underlying principles including cell autonomous ability, soluble communication, cell-cell adhesion, cell-matrix adhesion, and integrating these determinants of migration on molecular scaffolds. The current challenge is to implement meaningful and sustainable inhibition of metastasis by developing clinically viable disruption of molecular targets that control these fundamental capabilities. PMID:21664937

  20. Structural characterization of ultrathin Cr-doped ITO layers deposited by double-target pulsed laser ablation

    NASA Astrophysics Data System (ADS)

    Cesaria, Maura; Caricato, Anna Paola; Leggieri, Gilberto; Luches, Armando; Martino, Maurizio; Maruccio, Giuseppe; Catalano, Massimo; Grazia Manera, Maria; Rella, Roberto; Taurino, Antonietta

    2011-09-01

    In this paper we report on the growth and structural characterization of very thin (20 nm) Cr-doped ITO films, deposited at room temperature by double-target pulsed laser ablation on amorphous silica substrates. The role of Cr atoms in the ITO matrix is carefully investigated with increasing doping content by transmission electron microscopy (TEM). Selected-area electron diffraction, conventional bright field and dark field as well as high-resolution TEM analyses, and energy dispersive x-ray spectroscopy demonstrate that (i) crystallization features occur despite the low growth temperature and small thickness, (ii) no chromium or chromium oxide secondary phases are detectable, regardless of the film doping levels, (iii) the films crystallize as crystalline flakes forming large-angle grain boundaries; (iv) the observed flakes consist of crystalline planes with local bending of the crystal lattice. Thickness and compositional information about the films are obtained by Rutherford back-scattering spectrometry. Results are discussed by considering the combined effects of growth temperature, smaller ionic radius of the Cr cation compared with the trivalent In ion, doping level, film thickness, the double-target doping technique and peculiarities of the pulsed laser deposition method.

  1. Cytotoxic T cells use mechanical force to potentiate target cell killing

    PubMed Central

    Basu, Roshni; Whitlock, Benjamin M.; Husson, Julien; Le Floc’h, Audrey; Jin, Weiyang; Oyler-Yaniv, Alon; Dotiwala, Farokh; Giannone, Gregory; Hivroz, Claire; Biais, Nicolas; Lieberman, Judy; Kam, Lance C.; Huse, Morgan

    2016-01-01

    SUMMARY The immunological synapse formed between a cytotoxic T lymphocyte (CTL) and an infected or transformed target cell is a physically active structure capable of exerting mechanical force. Here, we investigated whether synaptic forces promote the destruction of target cells. CTLs kill by secreting toxic proteases and the pore forming protein perforin into the synapse. Biophysical experiments revealed a striking correlation between the magnitude of force exertion across the synapse and the speed of perforin pore formation on the target cell, implying that force potentiates cytotoxicity by enhancing perforin activity. Consistent with this interpretation, we found that increasing target cell tension augmented pore formation by perforin and killing by CTLs. Our data also indicate that CTLs coordinate perforin release and force exertion in space and time. These results reveal an unappreciated physical dimension to lymphocyte function and demonstrate that cells use mechanical forces to control the activity of outgoing chemical signals. PMID:26924577

  2. Cytotoxic T Cells Use Mechanical Force to Potentiate Target Cell Killing.

    PubMed

    Basu, Roshni; Whitlock, Benjamin M; Husson, Julien; Le Floc'h, Audrey; Jin, Weiyang; Oyler-Yaniv, Alon; Dotiwala, Farokh; Giannone, Gregory; Hivroz, Claire; Biais, Nicolas; Lieberman, Judy; Kam, Lance C; Huse, Morgan

    2016-03-24

    The immunological synapse formed between a cytotoxic T lymphocyte (CTL) and an infected or transformed target cell is a physically active structure capable of exerting mechanical force. Here, we investigated whether synaptic forces promote the destruction of target cells. CTLs kill by secreting toxic proteases and the pore forming protein perforin into the synapse. Biophysical experiments revealed a striking correlation between the magnitude of force exertion across the synapse and the speed of perforin pore formation on the target cell, implying that force potentiates cytotoxicity by enhancing perforin activity. Consistent with this interpretation, we found that increasing target cell tension augmented pore formation by perforin and killing by CTLs. Our data also indicate that CTLs coordinate perforin release and force exertion in space and time. These results reveal an unappreciated physical dimension to lymphocyte function and demonstrate that cells use mechanical forces to control the activity of outgoing chemical signals. Copyright © 2016 Elsevier Inc. All rights reserved.

  3. Isochoric heating of low Z solid targets with sub 10 fs laser pulses

    NASA Astrophysics Data System (ADS)

    Osterholz, Jens

    2004-11-01

    The investigation of high density plasmas plays an important role for astrophysics, inertial confinement fusion and x-ray lasers. Therefore the generation of dense plasmas with ultra-intense laser pulses is a field of enormous topical interest. An upper limit of the maximum plasma density that can be achieved with this method, however, occurs due to the formation of a preplasma and the expansion of the plasma during the interaction [1,2]. Here we describe a novel approach that is based on a laser system that generates sub 10 fs pulses with a low prepulse energy. Isochoric heating is demonstrated with small Z solid targets. Time integrated XUV spectroscopy is used to investigate K-shell emission from the plasma. In the spectra, only the Ly α and He α lines are observed, whereas transitions from orbitals with principal quantum numbers n > 2 are not present. This series limit is explained by pressure ionisation in the dense plasma. The XUV spectra were simulated by two different models [3]. The first calculates the effect of pressure ionisation and the second calculates the line intensity ratios. Preliminary calculations suggest that the plasma density of the emitting region is close to solid density with an electron temperature of about 100eV. We conclude that our laser system is well suited for isochoric heating of solid targets and an efficient transfer of the laser energy to the dense region of the target is possible. In cooperation with: T. Fischer, F. Brandl, G. Pretzler and O. Willi, Heinrich-Heine-University Duesseldorf, Germany, S. J. Rose, University of Oxford, United Kingdom [1] D. Riley et al., PRL 69, 3739 (1992). [2] A. Saemann et al., PRL 82, 4843 (1999). [3] S. J. Rose, J Phys B: Atom Molec Opt Phys, 25, 1667 (1992), 31, 2129 (1998).

  4. Development of pulsed processes for the manufacture of solar cells

    NASA Technical Reports Server (NTRS)

    Minnucci, J. A.

    1978-01-01

    The results of a 1-year program to develop the processes required for low-energy ion implantation for the automated production of silicon solar cells are described. The program included: (1) demonstrating state-of-the-art ion implantation equipment and designing an automated ion implanter, (2) making efforts to improve the performance of ion-implanted solar cells to 16.5 percent AM1, (3) developing a model of the pulse annealing process used in solar cell production, and (4) preparing an economic analysis of the process costs of ion implantation.

  5. Cell-to-Cell Transmission Can Overcome Multiple Donor and Target Cell Barriers Imposed on Cell-Free HIV

    PubMed Central

    Ilinskaya, Anna; Dorjbal, Batsukh; Truong, Rosaline; Derse, David; Uchil, Pradeep D.; Heidecker, Gisela; Mothes, Walther

    2013-01-01

    Virus transmission can occur either by a cell-free mode through the extracellular space or by cell-to-cell transmission involving direct cell-to-cell contact. The factors that determine whether a virus spreads by either pathway are poorly understood. Here, we assessed the relative contribution of cell-free and cell-to-cell transmission to the spreading of the human immunodeficiency virus (HIV). We demonstrate that HIV can spread by a cell-free pathway if all the steps of the viral replication cycle are efficiently supported in highly permissive cells. However, when the cell-free path was systematically hindered at various steps, HIV transmission became contact-dependent. Cell-to-cell transmission overcame barriers introduced in the donor cell at the level of gene expression and surface retention by the restriction factor tetherin. Moreover, neutralizing antibodies that efficiently inhibit cell-free HIV were less effective against cell-to-cell transmitted virus. HIV cell-to-cell transmission also efficiently infected target T cells that were relatively poorly susceptible to cell-free HIV. Importantly, we demonstrate that the donor and target cell types influence critically the extent by which cell-to-cell transmission can overcome each barrier. Mechanistically, cell-to-cell transmission promoted HIV spread to more cells and infected target cells with a higher proviral content than observed for cell-free virus. Our data demonstrate that the frequently observed contact-dependent spread of HIV is the result of specific features in donor and target cell types, thus offering an explanation for conflicting reports on the extent of cell-to-cell transmission of HIV. PMID:23308151

  6. First PIC simulations modeling the interaction of ultra-intense lasers with sub-micron, liquid crystal targets

    NASA Astrophysics Data System (ADS)

    McMahon, Matthew; Poole, Patrick; Willis, Christopher; Andereck, David; Schumacher, Douglass

    2014-10-01

    We recently introduced liquid crystal films as on-demand, variable thickness (50-5000 nanometers), low cost targets for intense laser experiments. Here we present the first particle-in-cell (PIC) simulations of short pulse laser excitation of liquid crystal targets treating Scarlet (OSU) class lasers using the PIC code LSP. In order to accurately model the target evolution, a low starting temperature and field ionization model are employed. This is essential as large starting temperatures, often used to achieve large Debye lengths, lead to expansion of the target causing significant reduction of the target density before the laser pulse can interact. We also present an investigation of the modification of laser pulses by very thin targets. This work was supported by the DARPA PULSE program through a grant from ARMDEC, by the US Department of Energy under Contract No. DE-NA0001976, and allocations of computing time from the Ohio Supercomputing Center.

  7. High-speed precise cell patterning by pulsed electrohydrodynamic jet printing

    NASA Astrophysics Data System (ADS)

    Makaev, A. V.; Mingaliev, E. A.; Karpov, V. R.; Zubarev, I. V.; Shur, V. Ya; El'kina, O. S.

    2017-10-01

    The generation of micro-droplets of nutrient medium with living cells by pulsed electrohydrodynamic printing has been studied. In-situ visualization by high-speed camera made it possible to measure the characteristic times of droplet generation process and to determine the optimal printing parameters. Maximal frequency of stable generation was achieved at 700 Hz. This technique was applied successfully for drop-on-demand printing of culture medium with live HeLa cells and yeasts.

  8. Single-Cell Droplet Microfluidic Screening for Antibodies Specifically Binding to Target Cells.

    PubMed

    Shembekar, Nachiket; Hu, Hongxing; Eustace, David; Merten, Christoph A

    2018-02-20

    Monoclonal antibodies are a main player in modern drug discovery. Many antibody screening formats exist, each with specific advantages and limitations. Nonetheless, it remains challenging to screen antibodies for the binding of cell-surface receptors (the most important class of all drug targets) or for the binding to target cells rather than purified proteins. Here, we present a high-throughput droplet microfluidics approach employing dual-color normalized fluorescence readout to detect antibody binding. This enables us to obtain quantitative data on target cell recognition, using as little as 33 fg of IgG per assay. Starting with an excess of hybridoma cells releasing unspecific antibodies, individual clones secreting specific binders (of target cells co-encapsulated into droplets) could be enriched 220-fold after sorting 80,000 clones in a single experiment. This opens the way for therapeutic antibody discovery, especially since the single-cell approach is in principle also applicable to primary human plasma cells. Copyright © 2018 The Authors. Published by Elsevier Inc. All rights reserved.

  9. Toward investigating changes in cell mechanoelastic properties in response to nanosecond pulsed electric fields

    NASA Astrophysics Data System (ADS)

    Coker, Zachary; Troyanova-Wood, Maria; Traverso, Andrew; Meng, Zhaokai; Ballmann, Charles; Petrov, Georgi; Ibey, Bennett L.; Yakovlev, Vladislav

    2017-02-01

    Nanosecond electric pulses (nsEPs) are known to cause a variety of effects on mammalian cells, ranging from destabilization of cell membranes to changes in cytoskeleton and elastic moduli. Measurement of a cells mechanoelastic properties have previously been limited to only invasive and destructive techniques such as atomic force microscopy or application of optical tweezers. However, due to recent advances, Brillouin spectroscopy has now become viable as a non-contact, non-invasive method for measuring these properties in cells and other materials. Here, we present progress toward applying Brillouin spectroscopy using a unique microscopy system for measuring changes in CHO-K1 cells when exposed to nsEPs of 600ns pulse duration with intensity of 50kV/cm. Successful measurement of mechanoelastic changes in these cells will demonstrate Brillouin spectroscopy as a viable method for measuring changes in elastic properties of other cells and living organisms.

  10. Pulsed Magnetic Field System for Magnetized Target Experiments at the National Ignition Facility

    NASA Astrophysics Data System (ADS)

    Rhodes, M. A.; Solberg, J. M.; Logan, B. G.; Perkins, L. J.

    2014-10-01

    High-magnitude magnetic fields applied to inertially confined targets may improve fusion yield and enable basic science applications. We discuss the development of a pulsed magnetic field system for NIF with the goal of applying 10--70 T to various NIF targets. While the driver may be little more than a spark-gap switched capacitor, numerous complex challenges exist in fielding such a system on NIF. The coil surrounding the metallic hohlraum drives induced current in the hohlraum wall. Both the coil and hohlraum wall must survive ohmic heating and J × B forces for several microseconds. Pulsed power must couple to the coil in the NIF environment. The system must not cause late-time optics damage due to debris. There is very limited volume for the driver in a NIF Diagnostic Instrument Manipulator (DIM). We are modeling the coil and hohlraum MHD effects with the LLNL code, ALE3D. However, the simulations lack complete and accurate data for all the required thermo-physical material properties over the expected range of temperatures (below vaporization) and pressures. Therefore, substantial experimental development is planned in the coming year. We present coil and hohlraum simulations results, overall system design, and progress towards an operational prototype test-stand. LLNL is operated by LLNS, LLC, for the U.S. D.O.E., NNSA under Contract DE-AC52-07NA27344. This work was supported by LLNL LDRD 14-ER-028.

  11. Electromagnetic Pulses Generated From Laser Target Interactions at Shenguang II Laser Facility

    NASA Astrophysics Data System (ADS)

    Yang, Jinwen; Li, Tingshuai; Yi, Tao; Wang, Chuanke; Yang, Ming; Yang, Weiming; Liu, Shenye; Jiang, Shaoen; Ding, Yongkun

    2016-10-01

    Significant electromagnetic pulses (EMP) can be generated by the intensive laser irradiating solid targets in inertial confinement fusion (ICF). To evaluate the EMP intensity and distribution in and outside the laser chamber, we designed and fabricated a discone antenna with ultra-wide bands of over 10 GHz. The return loss (S11 parameter) of this antenna was below -10 dB and could even achieve under -30 dB at 3.1 GHz. The EMP intensity in this study at 80 cm and 40 cm away from the target chamber center (TCC) reached 400 kV/m and 2000 kV/m. The current results are expected to offer preliminary information to study physics regarding laser plasma interactions and will also lay experimental foundation for EMI shielding design to protect various diagnostics. supported by the Fundamental Research Funds for the Central Universities of China (No. ZYGX2015J108) and National Natural Science Foundation of China (Nos. 11575166 and 51581140)

  12. Efficient Intracellular Delivery of Molecules with High Cell Viability Using Nanosecond-Pulsed Laser-Activated Carbon Nanoparticles

    PubMed Central

    2015-01-01

    Conventional physical and chemical methods that efficiently deliver molecules into cells are often associated with low cell viability. In this study, we evaluated the cellular effects of carbon nanoparticles believed to emit photoacoustic waves due to nanosecond-pulse laser activation to test the hypothesis that this method could achieve efficient intracellular delivery while maintaining high cell viability. Suspensions of DU145 human prostate carcinoma cells, carbon black (CB) nanoparticles, and calcein were exposed to 5–9 ns long laser pulses of near-infrared (1064 nm wavelength) light and then analyzed by flow cytometry for intracellular uptake of calcein and cell viability by propidium iodide staining. We found that intracellular uptake increased and in some cases saturated at high levels with only small losses in cell viability as a result of increasing laser fluence, laser exposure time, and as a unifying parameter, the total laser energy. Changing interpulse spacing between 0.1 and 10 s intervals showed no significant change in bioeffects, suggesting that the effects of each pulse were independent when spaced by at least 0.1 s intervals. Pretreatment of CB nanoparticles to intense laser exposure followed by mixing with cells also had no significant effect on uptake or viability. Similar uptake and viability were seen when CB nanoparticles were substituted with India ink, when DU145 cells were substituted with H9c2 rat cardiomyoblast cells, and when calcein was substituted with FITC-dextran. The best laser exposure conditions tested led to 88% of cells with intracellular uptake and close to 100% viability, indicating that nanosecond-pulse laser-activated carbon nanoparticles can achieve efficient intracellular delivery while maintaining high cell viability. PMID:24547946

  13. Experiments and PIC simulations on liquid crystal plasma mirrors for pulse contrast enhancement

    NASA Astrophysics Data System (ADS)

    Cochran, G. E.; Poole, P. L.; Krygier, A.; Foster, P. S.; Scott, G. G.; Wilson, L. A.; Bailey, J.; Bourgeois, N.; Hernandez-Gomez, C.; Heery, R.; Purcell, J.; Neely, D.; Rajeev, P. P.; Freeman, R. R.; Schumacher, D. W.

    2016-10-01

    High pulse contrast is crucial for performing many experiments on high intensity lasers in order to minimize modification of the target surface by pre-pulse. This is often achieved through the use of solid dielectric plasma mirrors which can limit laser shot rates. Liquid crystal films, originally developed as variable thickness ion acceleration targets, have been demonstrated as effective plasma mirrors for pulse cleaning, reaching peak reflectivities over 70%. These films were used as plasma mirrors in an ion acceleration experiment on the Scarlet laser and the resultant increase in peak proton energy and change in acceleration direction will be discussed. Also presented here are novel 2D3V, LSP particle-in-cell simulations of dielectric plasma mirror operation. By including multiphoton ionization and dimensionality corrections, an excellent match to experiment is obtained over 4 decades in intensity. Analysis of pulse shortening and plasma critical surface behavior in these simulations will be discussed. Formation of thin films at 1.5 Hz will also be presented. Performed with support from the DARPA PULSE program through AMRDEC, from NNSA, and from OSC.

  14. In situ single cell detection via microfluidic magnetic bead assay.

    PubMed

    Liu, Fan; Kc, Pawan; Zhang, Ge; Zhe, Jiang

    2017-01-01

    We present a single cell detection device based on magnetic bead assay and micro Coulter counters. This device consists of two successive micro Coulter counters, coupled with a high gradient magnetic field generated by an external magnet. The device can identify single cells in terms of the transit time difference of the cell through the two micro Coulter counters. Target cells are conjugated with magnetic beads via specific antibody and antigen binding. A target cell traveling through the two Coulter counters interacts with the magnetic field, and have a longer transit time at the 1st counter than that at the 2nd counter. In comparison, a non-target cell has no interaction with the magnetic field, and hence has nearly the same transit times through the two counters. Each cell passing through the two counters generates two consecutive voltage pulses one after the other; the pulse widths and magnitudes indicating the cell's transit times through the counters and the cell's size respectively. Thus, by measuring the pulse widths (transit times) of each cell through the two counters, each single target cell can be differentiated from non-target cells even if they have similar sizes. We experimentally proved that the target human umbilical vein endothelial cells (HUVECs) and non-target rat adipose-derived stem cells (rASCs) have significant different transit time distribution, from which we can determine the recognition regions for both cell groups quantitatively. We further demonstrated that within a mixed cell population of rASCs and HUVECs, HUVECs can be detected in situ and the measured HUVECs ratios agree well with the pre-set ratios. With the simple device structure and easy sample preparation, this method is expected to enable single cell detection in a continuous flow and can be applied to facilitate general cell detection applications such as stem cell identification and enumeration.

  15. Enhanced Monitoring of Nanosecond Electric Pulse-Evoked Membrane Conductance Changes in Whole-Cell Patch Clamp Experiments.

    PubMed

    Yoon, Jihwan; Leblanc, Normand; Zaklit, Josette; Vernier, P Thomas; Chatterjee, Indira; Craviso, Gale L

    2016-10-01

    Patch clamp electrophysiology serves as a powerful method for studying changes in plasma membrane ion conductance induced by externally applied high-intensity nanosecond electric pulses (NEPs). This paper describes an enhanced monitoring technique that minimizes the length of time between pulse exposure and data recording in a patch-clamped excitable cell. Whole-cell membrane currents were continuously recorded up to 11 ms before and resumed 8 ms after delivery of a 5-ns, 6 MV/m pulse by a pair of tungsten rod electrodes to a patched adrenal chromaffin cell maintained at a holding potential of -70 mV. This timing was achieved by two sets of relay switches. One set was used to disconnect the patch pipette electrode from the pre-amplifier and connect it to a battery to maintain membrane potential at -70 mV, and also to disconnect the reference electrode from the amplifier. The other set was used to disconnect the electrodes from the pulse generator until the time of NEP/sham exposure. The sequence and timing of both sets of relays were computer-controlled. Using this procedure, we observed that a 5-ns pulse induced an instantaneous inward current that decayed exponentially over the course of several minutes, that a second pulse induced a similar response, and that the current was carried, at least in part, by Na + . This approach for characterizing ion conductance changes in an excitable cell in response to NEPs will yield information essential for assessing the potential use of NEP stimulation for therapeutic applications.

  16. Ground-echo characteristics for a ground-target pulse-Doppler radar fuze of high duty ratio

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    Williams, C.S.

    1973-11-21

    From Tri-service electronic fuse symposium; Washington, District of Columbia, USA (26 Nov 1973). A pulse-Doppler radar fuze for use against ground targets at high burst heights can operate at low peak power provided a high duty ratio is used. The high duty ratio brings about ambiguous ground return that is prevented from firing the fuze by randomly coding the phase of the transmitted pulses. This causes the ambiguous return to appear as random noise. This paper provides formulas for the calculation of the clutter-noise power density and of the signal power so that the performance of the radar can bemore » determined. The paper also discusses the myth of decorrelation'' that is alleged to destroy the transmittedphase modulation in the echo and so make it useless. (auth)« less

  17. EFFECTS OF LASER RADIATION ON MATTER. LASER PLASMA: Generation of currents and propagation of plasma fronts in the case of two-pulse interaction with a target in air

    NASA Astrophysics Data System (ADS)

    Barkhudarov, É. M.; Gelashvili, G. V.; Gumberidze, G. G.; Taktakishvili, M. I.

    1990-06-01

    An investigation was made of the enhancement in the efficiency of generation of currents when a target in air was subjected to two consecutive CO2 laser radiation pulses. Preliminary interaction with a low-energy (1.5-5 J) pulse increased by more than one order of magnitude the currents generated by the second pulse and this was true in a wide range of energies of the latter pulse. The energy conversion efficiency was practically unaffected. The results were in qualitative agreement with the proposed pattern of plasma formation and propagation of shock waves near a target.

  18. Generic Sensor Modeling Using Pulse Method

    NASA Technical Reports Server (NTRS)

    Helder, Dennis L.; Choi, Taeyoung

    2005-01-01

    Recent development of high spatial resolution satellites such as IKONOS, Quickbird and Orbview enable observation of the Earth's surface with sub-meter resolution. Compared to the 30 meter resolution of Landsat 5 TM, the amount of information in the output image was dramatically increased. In this era of high spatial resolution, the estimation of spatial quality of images is gaining attention. Historically, the Modulation Transfer Function (MTF) concept has been used to estimate an imaging system's spatial quality. Sometimes classified by target shapes, various methods were developed in laboratory environment utilizing sinusoidal inputs, periodic bar patterns and narrow slits. On-orbit sensor MTF estimation was performed on 30-meter GSD Landsat4 Thematic Mapper (TM) data from the bridge pulse target as a pulse input . Because of a high resolution sensor s small Ground Sampling Distance (GSD), reasonably sized man-made edge, pulse, and impulse targets can be deployed on a uniform grassy area with accurate control of ground targets using tarps and convex mirrors. All the previous work cited calculated MTF without testing the MTF estimator's performance. In previous report, a numerical generic sensor model had been developed to simulate and improve the performance of on-orbit MTF estimating techniques. Results from the previous sensor modeling report that have been incorporated into standard MTF estimation work include Fermi edge detection and the newly developed 4th order modified Savitzky-Golay (MSG) interpolation technique. Noise sensitivity had been studied by performing simulations on known noise sources and a sensor model. Extensive investigation was done to characterize multi-resolution ground noise. Finally, angle simulation was tested by using synthetic pulse targets with angles from 2 to 15 degrees, several brightness levels, and different noise levels from both ground targets and imaging system. As a continuing research activity using the developed sensor

  19. Two-dimensional particle-in-cell plasma source ion implantation of a prolate spheroid target

    NASA Astrophysics Data System (ADS)

    Liu, Cheng-Sen; Han, Hong-Ying; Peng, Xiao-Qing; Chang, Ye; Wang, De-Zhen

    2010-03-01

    A two-dimensional particle-in-cell simulation is used to study the time-dependent evolution of the sheath surrounding a prolate spheroid target during a high voltage pulse in plasma source ion implantation. Our study shows that the potential contour lines pack more closely in the plasma sheath near the vertex of the major axis, i.e. where a thinner sheath is formed, and a non-uniform total ion dose distribution is incident along the surface of the prolate spheroid target due to the focusing of ions by the potential structure. Ion focusing takes place not only at the vertex of the major axis, where dense potential contour lines exist, but also at the vertex of the minor axis, where sparse contour lines exist. This results in two peaks of the received ion dose, locating at the vertices of the major and minor axes of the prolate spheroid target, and an ion dose valley, staying always between the vertices, rather than at the vertex of the minor axis.

  20. Clinical-scale laser-based scanning and processing of live cells: selective photothermal killing of fluorescent tumor targets for autologous stem cell transplantation

    NASA Astrophysics Data System (ADS)

    Koller, Manfred R.; Hanania, Elie G.; Eisfeld, Timothy; O'Neal, Robert A.; Khovananth, Kevin M.; Palsson, Bernhard O.

    2001-04-01

    High-dose chemotherapy, followed by autologous hematopoietic stem cell (HSC) transplantation, is widely used for the treatment of cancer. However, contaminating tumor cells within HSC harvests continue to be of major concern since re-infused tumor cells have proven to contribute to disease relapse. Many tumor purging methods have been evaluated, but all leave detectable tumor cells in the transplant and result in significant loss of HSCs. These shortcomings cause engraftment delays and compromise the therapeutic value of purging. A novel approach integrating automated scanning cytometry, image analysis, and selective laser-induced killing of labeled cells within a cell mixture is described here. Non-Hodgkin's lymphoma (NHL) cells were spiked into cell mixtures, and fluorochrome-conjugated antibodies were used to label tumor cells within the mixture. Cells were then allowed to settle on a surface, and as the surface was scanned with a fluorescence excitation source, a laser pulse was fired at every detected tumor cell using high-speed beam steering mirrors. Tumor cells were selectively killed with little effect on adjacent non-target cells, demonstrating the feasibility of this automated cell processing approach. This technology has many potential research and clinical applications, one example of which is tumor cell purging for autologous HSC transplantation.

  1. Controversies in targeted therapy of adult T cell leukemia/lymphoma: ON target or OFF target effects?

    PubMed

    Nasr, Rihab; El Hajj, Hiba; Kfoury, Youmna; de Thé, Hugues; Hermine, Olivier; Bazarbachi, Ali

    2011-06-01

    Adult T cell leukemia/lymphoma (ATL) represents an ideal model for targeted therapy because of intrinsic chemo-resistance of ATL cells and the presence of two well identified targets: the HTLV-I retrovirus and the viral oncoprotein Tax. The combination of zidovudine (AZT) and interferon-alpha (IFN) has a dramatic impact on survival of ATL patients. Although the mechanism of action remains unclear, arguments in favor or against a direct antiviral effect will be discussed. Yet, most patients relapse and alternative therapies are mandatory. IFN and arsenic trioxide induce Tax proteolysis, synergize to induce apoptosis in ATL cells and cure Tax-driven ATL in mice through specific targeting of leukemia initiating cell activity. These results provide a biological basis for the clinical success of arsenic/IFN/AZT therapy in ATL patients and suggest that both extinction of viral replication (AZT) and Tax degradation (arsenic/IFN) are needed to cure ATL.

  2. Ion acceleration enhanced by target ablation

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    Zhao, S.; State Key Laboratory of Nuclear Physics and Technology, and Key Lab of HEDPS, CAPT, Peking University, Beijing 100871; Institute of Radiation, Helmholtz-Zentrum Dresden-Rossendorf, 01314 Dresden

    2015-07-15

    Laser proton acceleration can be enhanced by using target ablation, due to the energetic electrons generated in the ablation preplasma. When the ablation pulse matches main pulse, the enhancement gets optimized because the electrons' energy density is highest. A scaling law between the ablation pulse and main pulse is confirmed by the simulation, showing that for given CPA pulse and target, proton energy improvement can be achieved several times by adjusting the target ablation.

  3. In situ single cell detection via microfluidic magnetic bead assay

    PubMed Central

    KC, Pawan; Zhang, Ge; Zhe, Jiang

    2017-01-01

    We present a single cell detection device based on magnetic bead assay and micro Coulter counters. This device consists of two successive micro Coulter counters, coupled with a high gradient magnetic field generated by an external magnet. The device can identify single cells in terms of the transit time difference of the cell through the two micro Coulter counters. Target cells are conjugated with magnetic beads via specific antibody and antigen binding. A target cell traveling through the two Coulter counters interacts with the magnetic field, and have a longer transit time at the 1st counter than that at the 2nd counter. In comparison, a non-target cell has no interaction with the magnetic field, and hence has nearly the same transit times through the two counters. Each cell passing through the two counters generates two consecutive voltage pulses one after the other; the pulse widths and magnitudes indicating the cell’s transit times through the counters and the cell’s size respectively. Thus, by measuring the pulse widths (transit times) of each cell through the two counters, each single target cell can be differentiated from non-target cells even if they have similar sizes. We experimentally proved that the target human umbilical vein endothelial cells (HUVECs) and non-target rat adipose-derived stem cells (rASCs) have significant different transit time distribution, from which we can determine the recognition regions for both cell groups quantitatively. We further demonstrated that within a mixed cell population of rASCs and HUVECs, HUVECs can be detected in situ and the measured HUVECs ratios agree well with the pre-set ratios. With the simple device structure and easy sample preparation, this method is expected to enable single cell detection in a continuous flow and can be applied to facilitate general cell detection applications such as stem cell identification and enumeration. PMID:28222140

  4. Antibody-targeted interleukin 2 stimulates T-cell killing of autologous tumor cells.

    PubMed Central

    Gillies, S D; Reilly, E B; Lo, K M; Reisfeld, R A

    1992-01-01

    A genetically engineered fusion protein consisting of a chimeric anti-ganglioside GD2 antibody (ch14.18) and interleukin 2 (IL2) was tested for its ability to enhance the killing of autologous GD2-expressing melanoma target cells by a tumor-infiltrating lymphocyte line (660 TIL). The fusion of IL2 to the carboxyl terminus of the immunoglobulin heavy chain did not reduce IL2 activity as measured in a standard proliferation assay using either mouse or human T-cell lines. Antigen-binding activity was greater than that of the native chimeric antibody. The ability of resting 660 TIL cells to kill their autologous GD2-positive target cells was enhanced if the target cells were first coated with the fusion protein. This stimulation of killing was greater than that of uncoated cells in the presence of equivalent or higher concentrations of free IL2. Such antibody-cytokine fusion proteins may prove useful in targeting the biological effect of IL2 and other cytokines to tumor cells and in this way stimulate their immune destruction. Images PMID:1741398

  5. Modeling dynamic plasmas driven by ultraintense nano-focused x-ray laser pulses in solid iron targets

    NASA Astrophysics Data System (ADS)

    Royle, Ryan; Sentoku, Yasuhiko; Mancini, Roberto

    2017-10-01

    The hard x-ray free electron laser has proven to be a valuable tool for high energy density (HED) physics as it is able to produce well-characterized samples of HED matter at exactly solid density and homogeneous temperatures. However, if the x-ray pulses are focused to sub-micron spot sizes, where peak intensities can exceed 1020 W/cm2, the plasmas driven by sources of non-thermal photoelectrons and Auger electrons can be highly dynamic and so cannot be modeled by atomic kinetics or fluid codes. We apply the 2D/3D particle-in-cell code, PICLS-which has been extended with numerous physics models to enable the simulation of XFEL-driven plasmas-to the modeling of such dynamic plasmas driven by nano-focused XFEL pulses in solid iron targets. In the case of the smallest focal spot investigated of just 100 nm in diameter, keV plasmas induce strong radial E-fields that accelerate keV ions radially as well as sheath fields that accelerate surface ions to hundreds of keV. The heated spot, which is initially larger than the laser spot due to the kinetic nature of the fast Auger electrons, expands as ion and electron waves propagate radially, leaving a low density region along the laser axis. This research was supported by the US DOE-OFES under Grant No. DE-SC0008827, the DOE-NNSA under Grant No. DE-NA0002075, and the JSPS KAKENHI under Grant No. JP15K21767.

  6. Observation of Gigawatt-Class THz Pulses from a Compact Laser-Driven Particle Accelerator

    NASA Astrophysics Data System (ADS)

    Gopal, A.; Herzer, S.; Schmidt, A.; Singh, P.; Reinhard, A.; Ziegler, W.; Brömmel, D.; Karmakar, A.; Gibbon, P.; Dillner, U.; May, T.; Meyer, H.-G.; Paulus, G. G.

    2013-08-01

    We report the observation of subpicosecond terahertz (T-ray) pulses with energies ≥460μJ from a laser-driven ion accelerator, thus rendering the peak power of the source higher even than that of state-of-the-art synchrotrons. Experiments were performed with intense laser pulses (up to 5×1019W/cm2) to irradiate thin metal foil targets. Ion spectra measured simultaneously showed a square law dependence of the T-ray yield on particle number. Two-dimensional particle-in-cell simulations show the presence of transient currents at the target rear surface which could be responsible for the strong T-ray emission.

  7. The influences of target properties and deposition times on pulsed laser deposited hydroxyapatite films

    NASA Astrophysics Data System (ADS)

    Bao, Quanhe; Chen, Chuanzhong; Wang, Diangang; Liu, Junming

    2008-11-01

    Hydroxyapatite films were produced by pulsed laser deposition from three kinds of hydroxyapatite targets and with different deposition times. A JXA-8800R electron probe microanalyzer (EPMA) with a Link ISIS300 energy spectrum analyzer was used to give the secondary electron image (SE) and determine the element composition of the films. The phases of thin film were analyzed by a D/max-γc X-ray diffractometer (XRD). The Fourier-transform infrared spectroscopy (FT-IR) was used to characterize the hydroxyl, phosphate and other functional groups. The results show that deposited films were amorphous which mainly composed of droplet-like particles and vibration of PO 43- groups. With the target sintering temperature deposition times increasing, the density of droplets is decreased. While with deposition times increasing, the density of droplets is increased. With the target sintering temperature and deposition time increasing, the ratio of Ca/P is increasing and higher than that of theoretical value of HA.

  8. Cell-targeted platinum nanoparticles and nanoparticle clusters.

    PubMed

    Papst, Stefanie; Brimble, Margaret A; Evans, Clive W; Verdon, Daniel J; Feisst, Vaughan; Dunbar, P Rod; Tilley, Richard D; Williams, David E

    2015-06-21

    Herein, we report the facile preparation of cell-targeted platinum nanoparticles (PtNPs), through the design of peptides that, as a single molecule added in small concentration during the synthesis, control the size of PtNP clusters during their growth, stabilise the PtNPs in aqueous suspension and enable the functionalisation of the PtNPs with a versatile range of cell-targeting ligands. Water-soluble PtNPs targeted respectively at blood group antigens and at integrin receptors are demonstrated.

  9. Laser plasma cryogenic target on translating substrate for generation of continuously repetitive EUV and soft X-ray pulses

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    Amano, Sho

    2014-06-15

    To generate continuously repetitive EUV and soft X-ray pulses with various wavelengths from laser-produced plasmas, a one-dimensionally translating substrate system with a closed He gas cryostat that can continuously supply various cryogenic targets for ∼10 Hz laser pulses has been developed. The system was successfully operated at a lowest temperature of 15 K and at a maximum up-down speed of 12 mm/s. Solid Ar, Kr, and Xe layers were formed, and their growth rates and the laser crater sizes on them were studied. By optimization of the operational parameters in accordance with our design rule, it was shown that stablemore » output power was achieved continuously from the plasma emission at frequencies of 1–10 Hz. The average soft X-ray and EUV powers obtained were 19 mW at 3.2 nm, 33 mW at 10.0 nm, and 66 mW at 10.8 nm, with 10% bandwidths, from the Ar, Kr, and Xe solid targets, respectively, with a laser power of 1 W. We will be able to achieve higher frequencies using a high beam quality laser that produces smaller craters, and can expect higher powers. Although only Ar, Kr, and Xe gases were tested in this study, the target system achieved a temperature of 15 K and can thus solidify almost all target gases, apart from H and He, and can continuously supply the solid target. The use of various target materials will enable expansion of the EUV and soft X-ray emission wavelength range.« less

  10. Resonant photoacoustic cell for pulsed laser analysis of gases at high temperature

    NASA Astrophysics Data System (ADS)

    Sorvajärvi, Tapio; Manninen, Albert; Toivonen, Juha; Saarela, Jaakko; Hernberg, Rolf

    2009-12-01

    A new approach to high temperature gas analysis by means of photoacoustic (PA) spectroscopy is presented. The transverse modes of the resonant PA cell were excited with a pulsed laser and detected with a microphone. Changes in the properties of the PA cell resulting from a varying temperature are discussed and considered when processing the PA signal. The feasibility of the proposed method was demonstrated by studying PA response from saturated vapor of potassium chloride (KCl) in the temperature range extending from 410 to 691 °C. The PA spectrum, the detection limit, and the signal saturation of KCl vapor are discussed. At 245 nm excitation wavelength and 300 μJ pulse energy, the achieved detection limit for KCl is 15 ppb.

  11. Pulsed-neutron monochromator

    DOEpatents

    Mook, H.A. Jr.

    1984-01-01

    In one aspect, the invention is an improved pulsed-neutron monochromator of the vibrated-crystal type. The monochromator is designed to provide neutron pulses which are characterized both by short duration and high density. A row of neutron-reflecting crystals is disposed in a neutron beam to reflect neutrons onto a common target. The crystals in the row define progressively larger neutron-scattering angles and are vibrated sequentially in descending order with respect to the size of their scattering angles, thus generating neutron pulses which arrive simultaneously at the target. Transducers are coupled to one end of the crystals to vibrate them in an essentially non-resonant mode. The transducers propagate transverse waves in the crystal which progress longitudinally therein. The waves are absorbed at the undriven ends of the crystals by damping material mounted thereon. In another aspect, the invention is a method for generating neutron pulses characterized by high intensity and short duration.

  12. Pulsed-neutron monochromator

    DOEpatents

    Mook, Jr., Herbert A.

    1985-01-01

    In one aspect, the invention is an improved pulsed-neutron monochromator of the vibrated-crystal type. The monochromator is designed to provide neutron pulses which are characterized both by short duration and high density. A row of neutron-reflecting crystals is disposed in a neutron beam to reflect neutrons onto a common target. The crystals in the row define progressively larger neutron-scattering angles and are vibrated sequentially in descending order with respect to the size of their scattering angles, thus generating neutron pulses which arrive simultaneously at the target. Transducers are coupled to one end of the crystals to vibrate them in an essentially non-resonant mode. The transducers propagate transverse waves in the crystal which progress longitudinally therein. The wave are absorbed at the undriven ends of the crystals by damping material mounted thereon. In another aspect, the invention is a method for generating neutron pulses characterized by high intensity and short duration.

  13. Pulse laser ablation of Au, Ag, and Cu metal targets in liquid for nanoparticle production

    NASA Astrophysics Data System (ADS)

    Herbani, Y.; Irmaniar; Nasution, R. S.; Mujtahid, F.; Masse, S.

    2018-03-01

    We have fabricated metal and oxide nanoparticles using pulse laser ablation of Au, Ag, and Cu metal targets immersed in water. While laser ablation of Au and Ag targets in water produced metal nanoparticles which were stable for a month even without any dispersant, we found CuO nanoparticles for Cu target due to rapid oxidation of Cu in water resulted in its poor stability. Au, Ag, and CuO nanoparticles production were barely identified by naked eyes for their distinctive colour of red, yellow, and dark green colloidal suspensions, respectively. It was also verified using UV-Vis spectrometer that Au, Ag, and CuO colloidal nanoparticles have their respective surface plasmon resonance at 520, 400, and 620 nm. TEM observation showed that particle sizes for all the fabricated nanoparticles were in the range of 20 – 40 nm with crystalline structures.

  14. Effects of front-surface target structures on properties of relativistic laser-plasma electrons.

    PubMed

    Jiang, S; Krygier, A G; Schumacher, D W; Akli, K U; Freeman, R R

    2014-01-01

    We report the results of a study of the role of prescribed geometrical structures on the front of a target in determining the energy and spatial distribution of relativistic laser-plasma electrons. Our three-dimensional particle-in-cell simulation studies apply to short-pulse, high-intensity laser pulses, and indicate that a judicious choice of target front-surface geometry provides the realistic possibility of greatly enhancing the yield of high-energy electrons while simultaneously confining the emission to narrow (<5°) angular cones.

  15. Pulse oximeter saturation target limits for preterm infants: a survey among European neonatal intensive care units.

    PubMed

    Huizing, Maurice J; Villamor-Martínez, Eduardo; Vento, Máximo; Villamor, Eduardo

    2017-01-01

    The optimum range of pulse oximeter oxygen saturation (SpO 2 ) for preterm infants remains controversial. Between November 2015 and February 2016, we conducted a web-based survey aimed to investigate the current and former practices on SpO 2 targets in European neonatal intensive care units (NICUs). We obtained valid responses from 193 NICUs, treating 8590 newborns ≤28 weeks per year, across 27 countries. Forty different saturation ranges were reported, ranging from 82-93 to 94-99%. The most frequently utilized SpO 2 ranges were 90-95% (28%), 88-95% (12%), 90-94% (5%), and 91-95% (5%). A total of 156 NICUs (81%) changed their SpO 2 limits over the last 10 years. The most frequently reported former limits were 88-92% (18%), 85-95% (9%), 88-93 (7%), and 85-92% (6%). The NICUs that increased their SpO 2 ranges expected to obtain a reduction in mortality. A 54% of the NICUs found the scientific evidence supporting their SpO 2 targeting policy strong or very strong. We detected a high degree of heterogeneity in pulse oximeter SpO 2 target limits across European NICUs. The currently used limits are 3 to 5% higher than the former limits, and the most extreme limits, such as lower below 85% or upper above 96%, have almost been abandoned. What is Known: • For preterm infants requiring supplemental oxygen, the optimum range of pulse oximeter oxygen saturation (SpO 2 ) to minimize organ damage, without causing hypoxic injury, remains controversial. What is New: • This survey highlights the lack of consensus regarding SpO 2 target limits for preterm infants among European neonatal intensive care units (NICUs). We detected 40 different SpO 2 ranges, and even the most frequently reported range (i.e., 90-95%) was used in only 28% of the 193 respondent NICUs. • A total of 156 NICUs (81%) changed their SpO 2 limits over the last 10 years. The currently used limits are 3 to 5% higher than the former limits, and the most extreme limits, such as lower below 85% or upper

  16. Cell perforation mediated by plasmonic bubbles generated by a single near infrared femtosecond laser pulse.

    PubMed

    Boutopoulos, Christos; Bergeron, Eric; Meunier, Michel

    2016-01-01

    We report on transient membrane perforation of living cancer cells using plasmonic gold nanoparticles (AuNPs) enhanced single near infrared (NIR) femtosecond (fs) laser pulse. Under optimized laser energy fluence, single pulse treatment (τ = 45 fs, λ = 800 nm) resulted in 77% cell perforation efficiency and 90% cell viability. Using dark field and ultrafast imaging, we demonstrated that the generation of submicron bubbles around the AuNPs is the necessary condition for the cell membrane perforation. AuNP clustering increased drastically the bubble generation efficiency, thus enabling an effective laser treatment using low energy dose in the NIR optical therapeutical window. © 2015 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  17. Controversies in Targeted Therapy of Adult T Cell Leukemia/Lymphoma: ON Target or OFF Target Effects?

    PubMed Central

    Nasr, Rihab; Hajj, Hiba El; Kfoury, Youmna; de Thé, Hugues; Hermine, Olivier; Bazarbachi, Ali

    2011-01-01

    Adult T cell leukemia/lymphoma (ATL) represents an ideal model for targeted therapy because of intrinsic chemo-resistance of ATL cells and the presence of two well identified targets: the HTLV-I retrovirus and the viral oncoprotein Tax. The combination of zidovudine (AZT) and interferon-alpha (IFN) has a dramatic impact on survival of ATL patients. Although the mechanism of action remains unclear, arguments in favor or against a direct antiviral effect will be discussed. Yet, most patients relapse and alternative therapies are mandatory. IFN and arsenic trioxide induce Tax proteolysis, synergize to induce apoptosis in ATL cells and cure Tax-driven ATL in mice through specific targeting of leukemia initiating cell activity. These results provide a biological basis for the clinical success of arsenic/IFN/AZT therapy in ATL patients and suggest that both extinction of viral replication (AZT) and Tax degradation (arsenic/IFN) are needed to cure ATL. PMID:21994752

  18. Harmonizing HeLa cell cytoskeleton behavior by multi-Ti oxide phased nanostructure synthesized through ultrashort pulsed laser

    PubMed Central

    Chinnakkannu Vijayakumar, Chandramouli; Venkatakrishnan, Krishnan; Tan, Bo

    2015-01-01

    Knowledge about cancer cell behavior on heterogeneous nanostructures is relevant for developing a distinct biomaterial that can actuate cancer cells. In this manuscript, we have demonstrated a harmonized approach of forming multi Ti-oxide phases in a nanostructure (MTOP nanostructure) for its unique cancer cell controlling behavior.Conventionally, single phases of TiO2 are used for targeted therapy and as drug carrier systems.In this research, we have shown a biomaterial that can control HeLa cells diligently using a combination of TiO, Ti3O and TiO2 phases when compared to fibroblast (NIH3T3) cells.MTOP-nanostructures are generated by varying the ionization energy in the vapor plume of the ultrashort pulse laser; this interaction with the material allows accurate tuning and composition of phases within the nanostructure. In addition, the lattice spacing of MTOP-nanostructures was analyzed as shown by HR-TEM investigations. An FESEM investigation of MTOP-nanostructures revealed a greater reduction of HeLa cells relative to fibroblast cells. Altered cell adhesion was followed by modulation of HeLa cell architecture with a significant reduction of actin stress fibers.The intricate combination of MTOP-nanostructures renders a biomaterial that can precisely alter HeLa cell but not fibroblast cell behavior, filling a void in the research for a biomaterial to modulate cancer cell behavior. PMID:26469886

  19. Harmonizing HeLa cell cytoskeleton behavior by multi-Ti oxide phased nanostructure synthesized through ultrashort pulsed laser

    NASA Astrophysics Data System (ADS)

    Chinnakkannu Vijayakumar, Chandramouli; Venkatakrishnan, Krishnan; Tan, Bo

    2015-10-01

    Knowledge about cancer cell behavior on heterogeneous nanostructures is relevant for developing a distinct biomaterial that can actuate cancer cells. In this manuscript, we have demonstrated a harmonized approach of forming multi Ti-oxide phases in a nanostructure (MTOP nanostructure) for its unique cancer cell controlling behavior.Conventionally, single phases of TiO2 are used for targeted therapy and as drug carrier systems.In this research, we have shown a biomaterial that can control HeLa cells diligently using a combination of TiO, Ti3O and TiO2 phases when compared to fibroblast (NIH3T3) cells.MTOP-nanostructures are generated by varying the ionization energy in the vapor plume of the ultrashort pulse laser; this interaction with the material allows accurate tuning and composition of phases within the nanostructure. In addition, the lattice spacing of MTOP-nanostructures was analyzed as shown by HR-TEM investigations. An FESEM investigation of MTOP-nanostructures revealed a greater reduction of HeLa cells relative to fibroblast cells. Altered cell adhesion was followed by modulation of HeLa cell architecture with a significant reduction of actin stress fibers.The intricate combination of MTOP-nanostructures renders a biomaterial that can precisely alter HeLa cell but not fibroblast cell behavior, filling a void in the research for a biomaterial to modulate cancer cell behavior.

  20. Microdosimetric study for nanosecond pulsed electric fields on a cell circuit model with nucleus.

    PubMed

    Denzi, Agnese; Merla, Caterina; Camilleri, Paola; Paffi, Alessandra; d'Inzeo, Guglielmo; Apollonio, Francesca; Liberti, Micaela

    2013-10-01

    Recently, scientific interest in electric pulses, always more intense and shorter and able to induce biological effects on both plasma and nuclear membranes, has greatly increased. Hence, microdosimetric models that include internal organelles like the nucleus have assumed increasing importance. In this work, a circuit model of the cell including the nucleus is proposed, which accounts for the dielectric dispersion of all cell compartments. The setup of the dielectric model of the nucleus is of fundamental importance in determining the transmembrane potential (TMP) induced on the nuclear membrane; here, this is demonstrated by comparing results for three different sets of nuclear dielectric properties present in the literature. The results have been compared, even including or disregarding the dielectric dispersion of the nucleus. The main differences have been found when using pulses shorter than 10 ns. This is due to the fact that the high spectral components of the shortest pulses are differently taken into account by the nuclear membrane transfer functions computed with and without nuclear dielectric dispersion. The shortest pulses are also the most effective in porating the intracellular structures, as confirmed by the time courses of the TMP calculated across the plasma and nuclear membranes. We show how dispersive nucleus models are unavoidable when dealing with pulses shorter than 10 ns because of the large spectral contents arriving above 100 MHz, i.e., over the typical relaxation frequencies of the dipolar mechanism of the molecules constituting the nuclear membrane and the subcellular cell compartments.

  1. Agricultural and Food Processing Applications of Pulsed Power Technology

    NASA Astrophysics Data System (ADS)

    Takaki, Koichi; Ihara, Satoshi

    Recent progress of agricultural and food processing applications of pulsed power is described in this paper. Repetitively operated compact pulsed power generators with a moderate peak power have been developed for the agricultural and the food processing applications. These applications are mainly based on biological effects and can be categorized as decontamination of air and liquid, germination promotion, inhabitation of saprophytes growth, extraction of juice from fruits and vegetables, and fertilization of liquid medium, etc. Types of pulsed power that have biological effects are caused with gas discharges, water discharges, and electromagnetic fields. The discharges yield free radicals, UV radiation, intense electric field, and shock waves. Biologically based applications of pulsed power are performed by selecting the type that gives the target objects the adequate result from among these agents or byproducts. For instance, intense electric fields form pores on the cell membrane, which is called electroporation, or influence the nuclei.

  2. Spatial and Temporal Confined Photothermolysis of Cancer Cells Mediated by Hollow Gold Nanospheres Targeted to Epidermal Growth Factor Receptors

    PubMed Central

    2018-01-01

    To date, a few studies have investigated the potential use of a short-pulsed laser in selective tumor cell destruction or its mechanism of cell killing. Computer simulation of the spatial and temporal profiles of temperature elevation after pulsed laser irradiation on an infinitesimal point source estimated that the temperature reached its highest point at ∼35 ns after a single 15 ns laser pulse. Moreover, temperature elevation was confined to a radius of sub-micrometer and returned to baseline within 100 ns. To investigate the effect of 15 ns laser pulses on A431 tumor cells, we conjugated hollow gold nanospheres (HAuNSs) to an antibody (C225) directed at the epithelial growth factor receptor. The resulting nanoparticles, C225-HAuNSs, bound to the cell membrane, internalized, and distributed throughout the cytoplasm, with some nanoparticles transported to the vicinity of the nuclear membrane. On using an optical microscope mounted to a tunable pulsed Ti:sapphire laser, rapid and extensive damage of live cancer cells was observed, whereas irradiation of A431 cells pretreated with nontargeted HAuNSs with a pulsed laser or pretreated with C225-HAuNSs with a continuous-wave laser-induced minimal cellular damage. Furthermore, after a single 15 ns laser pulse, C225-HAuNS-treated A431 cells cocultured with 3T3 fibroblasts showed signs of selective destruction. Thus, compared with a continuous-wave laser, shots of a short-pulsed laser were the most damaging to tumor cells that bound HAuNSs and generated the least heat to the surrounding environment. This mode of action by a short-pulsed laser on cancer cells (i.e., confined photothermolysis) may have potential applications in selective tumor cell destruction. PMID:29876540

  3. Spatial and Temporal Confined Photothermolysis of Cancer Cells Mediated by Hollow Gold Nanospheres Targeted to Epidermal Growth Factor Receptors.

    PubMed

    Ku, Geng; Huang, Qian; Wen, Xiaoxia; Ye, John; Piwnica-Worms, David; Li, Chun

    2018-05-31

    To date, a few studies have investigated the potential use of a short-pulsed laser in selective tumor cell destruction or its mechanism of cell killing. Computer simulation of the spatial and temporal profiles of temperature elevation after pulsed laser irradiation on an infinitesimal point source estimated that the temperature reached its highest point at ∼35 ns after a single 15 ns laser pulse. Moreover, temperature elevation was confined to a radius of sub-micrometer and returned to baseline within 100 ns. To investigate the effect of 15 ns laser pulses on A431 tumor cells, we conjugated hollow gold nanospheres (HAuNSs) to an antibody (C225) directed at the epithelial growth factor receptor. The resulting nanoparticles, C225-HAuNSs, bound to the cell membrane, internalized, and distributed throughout the cytoplasm, with some nanoparticles transported to the vicinity of the nuclear membrane. On using an optical microscope mounted to a tunable pulsed Ti:sapphire laser, rapid and extensive damage of live cancer cells was observed, whereas irradiation of A431 cells pretreated with nontargeted HAuNSs with a pulsed laser or pretreated with C225-HAuNSs with a continuous-wave laser-induced minimal cellular damage. Furthermore, after a single 15 ns laser pulse, C225-HAuNS-treated A431 cells cocultured with 3T3 fibroblasts showed signs of selective destruction. Thus, compared with a continuous-wave laser, shots of a short-pulsed laser were the most damaging to tumor cells that bound HAuNSs and generated the least heat to the surrounding environment. This mode of action by a short-pulsed laser on cancer cells (i.e., confined photothermolysis) may have potential applications in selective tumor cell destruction.

  4. Femtosecond laser pulses for chemical-free embryonic and mesenchymal stem cell differentiation

    NASA Astrophysics Data System (ADS)

    Mthunzi, Patience; Dholakia, Kishan; Gunn-Moore, Frank

    2011-10-01

    Owing to their self renewal and pluripotency properties, stem cells can efficiently advance current therapies in tissue regeneration and/or engineering. Under appropriate culture conditions in vitro, pluripotent stem cells can be primed to differentiate into any cell type some examples including neural, cardiac and blood cells. However, there still remains a pressing necessity to answer the biological questions concerning how stem cell renewal and how differentiation programs are operated and regulated at the genetic level. In stem cell research, an urgent requirement on experimental procedures allowing non-invasive, marker-free observation of growth, proliferation and stability of living stem cells under physiological conditions exists. Femtosecond (fs) laser pulses have been reported to non-invasively deliver exogenous materials, including foreign genetic species into both multipotent and pluripotent stem cells successfully. Through this multi-photon facilitated technique, directly administering fs laser pulses onto the cell plasma membrane induces transient submicrometer holes, thereby promoting cytosolic uptake of the surrounding extracellular matter. To display a chemical-free cell transfection procedure that utilises micro-litre scale volumes of reagents, we report for the first time on 70 % transfection efficiency in ES-E14TG2a cells using the enhanced green fluorescing protein (EGFP) DNA plasmid. We also show how varying the average power output during optical transfection influences cell viability, proliferation and cytotoxicity in embryonic stem cells. The impact of utilizing objective lenses of different numerical aperture (NA) on the optical transfection efficiency in ES-E14TG2a cells is presented. Finally, we report on embryonic and mesenchymal stem cell differentiation. The produced specialized cell types could thereafter be characterized and used for cell based therapies.

  5. PGE2 pulsing of murine bone marrow cells reduces migration of daughter monocytes/macrophages in vitro and in vivo

    PubMed Central

    McGonigle, Terence A.; Dwyer, Amy R.; Greenland, Eloise L.; Scott, Naomi M.; Keane, Kevin N.; Newsholme, Philip; Goodridge, Helen S.; Zon, Leonard I.; Pixley, Fiona J.; Hart, Prue H.

    2018-01-01

    Monocytes/macrophages differentiating from bone marrow (BM) cells pulsed for 2 hours at 37°C with a stabilized derivative of prostaglandin E2, 16,16-dimethyl PGE2 (dmPGE2), migrated less efficiently toward a chemoattractant than monocytes/macrophages differentiated from BM cells pulsed with vehicle. To confirm that the effect on BM cells was long lasting and to replicate human BM transplantation, chimeric mice were established with donor BM cells pulsed for 2 hours with dmPGE2 before injection into marrow-ablated congenic recipient mice. After 12 weeks, when high levels (90%) of engraftment were obtained, regenerated BM-derived monocytes/macrophages differentiating in vitro or in vivo migrated inefficiently toward the chemokines colony-stimulating factor-1 (CSF-1) and chemokine (C-C motif) ligand 2 (CCL2) or thioglycollate, respectively. Our results reveal long-lasting changes to progenitor cells of monocytes/macrophages by a 2-hour dmPGE2 pulse that, in turn, limits the migration of their daughter cells to chemoattractants and inflammatory mediators. PMID:28822771

  6. Noncontact microsurgery and delivery of substances into stem cells by means of femtosecond laser pulses

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    Il'ina, I V; Ovchinnikov, A V; Sitnikov, D S

    We have studied the efficiency of microsurgery of a cell membrane in mesenchymal stem cells and the posterior cell viability under the localised short-time action of femtosecond IR laser pulses aimed at noncontact delivery of specified substances into the cells. (extreme light fields and their applications)

  7. In-situ formation of solidified hydrogen thin-membrane targets using a pulse tube cryocooler

    NASA Astrophysics Data System (ADS)

    Astbury, S.; Bedacht, S.; Brummitt, P.; Carroll, D.; Clarke, R.; Crisp, S.; Hernandez-Gomez, C.; Holligan, P.; Hook, S.; Merchan, J. S.; Neely, D.; Ortner, A.; Rathbone, D.; Rice, P.; Schaumann, G.; Scott, G.; Spindloe, C.; Spurdle, S.; Tebartz, A.; Tomlinson, S.; Wagner, F.; Borghesi, M.; Roth, M.; Tolley, M. K.

    2016-04-01

    An account is given of the Central Laser Facility's work to produce a cryogenic hydrogen targetry system using a pulse tube cryocooler. Due to the increasing demand for low Z thin laser targets, CLF (in collaboration with TUD) have been developing a system which allows the production of solid hydrogen membranes by engineering a design which can achieve this remotely; enabling the gas injection, condensation and solidification of hydrogen without compromising the vacuum of the target chamber. A dynamic sealing mechanism was integrated which allows targets to be grown and then remotely exposed to open vacuum for laser interaction. Further research was conducted on the survivability of the cryogenic targets which concluded that a warm gas effect causes temperature spiking when exposing the solidified hydrogen to the outer vacuum. This effect was shown to be mitigated by improving the pumping capacity of the environment and reducing the minimum temperature obtainable on the target mount. This was achieved by developing a two-stage radiation shield encased with superinsulating blanketing; reducing the base temperature from 14 ± 0.5 K to 7.2 ± 0.2 K about the coldhead as well as improving temperature control stability following the installation of a high-performance temperature controller and sensor apparatus. The system was delivered experimentally and in July 2014 the first laser shots were taken upon hydrogen targets in the Vulcan TAP facility.

  8. Efficient energy absorption of intense ps-laser pulse into nanowire target

    NASA Astrophysics Data System (ADS)

    Habara, H.; Honda, S.; Katayama, M.; Sakagami, H.; Nagai, K.; Tanaka, K. A.

    2016-06-01

    The interaction between ultra-intense laser light and vertically aligned carbon nanotubes is investigated to demonstrate efficient laser-energy absorption in the ps laser-pulse regime. Results indicate a clear enhancement of the energy conversion from laser to energetic electrons and a simultaneously small plasma expansion on the surface of the target. A two-dimensional plasma particle calculation exhibits a high absorption through laser propagation deep into the nanotube array, even for a dense array whose structure is much smaller than the laser wavelength. The propagation leads to the radial expansion of plasma perpendicular to the nanotubes rather than to the front side. These features may contribute to fast ignition in inertial confinement fusion and laser particle acceleration, both of which require high current and small surface plasma simultaneously.

  9. Efficient energy absorption of intense ps-laser pulse into nanowire target

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    Habara, H.; Honda, S.; Katayama, M.

    The interaction between ultra-intense laser light and vertically aligned carbon nanotubes is investigated to demonstrate efficient laser-energy absorption in the ps laser-pulse regime. Results indicate a clear enhancement of the energy conversion from laser to energetic electrons and a simultaneously small plasma expansion on the surface of the target. A two-dimensional plasma particle calculation exhibits a high absorption through laser propagation deep into the nanotube array, even for a dense array whose structure is much smaller than the laser wavelength. The propagation leads to the radial expansion of plasma perpendicular to the nanotubes rather than to the front side. Thesemore » features may contribute to fast ignition in inertial confinement fusion and laser particle acceleration, both of which require high current and small surface plasma simultaneously.« less

  10. Timing of pulsed electromagnetic field stimulation does not affect the promotion of bone cell development.

    PubMed

    Hannay, Gwynne; Leavesley, David; Pearcy, Mark

    2005-12-01

    Pulsed electromagnetic field (PEMF) devices have been used clinically to promote the healing of surgically resistant fractures in vivo. However, there is a sparsity of data on how the timing of an applied PEMF effects the osteogenic cells that would be present within the fracture gap. The purpose of this study was to examine the response of osteoblast-like cells to a PEMF stimulus, mimicking that of a clinically available device, using four protocols for the timing of the stimulus. The PEMF signal consisted of a 5 ms pulse burst (containing 20 pulses) repeated at 15 Hz. Cultures of a human osteosarcoma cell line, SaOS-2, were exposed to the four timing protocols, each conducted over 3 days. Protocol one stimulated the cells for 8 h each day, protocol two stimulated the cells for 24 h on the first day, protocol three stimulated the cells for 24 h on the second day, and protocol four stimulated the cells for 24 h on the third day. Cells were seeded with either 25,000 or 50,000 cells/well (24-well cell culture plates). All assays showed reduced proliferation and increased differentiation (alkaline phosphatase activity) in the PEMF stimulated cultures compared with the control cultures, except for protocol four alkaline phosphatase measurements. No clear trend was observed between the four protocols; however this may be due to cell density. The results indicated that an osteoblast-like cell line is responsive to a 15 Hz PEMF stimulus, which will stimulate the cell line to into an increasing state of maturity. Bioelectromagnetics (c) 2005 Wiley-Liss, Inc

  11. Cell-permeable nanobodies for targeted immunolabelling and antigen manipulation in living cells

    NASA Astrophysics Data System (ADS)

    Herce, Henry D.; Schumacher, Dominik; Schneider, Anselm F. L.; Ludwig, Anne K.; Mann, Florian A.; Fillies, Marion; Kasper, Marc-André; Reinke, Stefan; Krause, Eberhard; Leonhardt, Heinrich; Cardoso, M. Cristina; Hackenberger, Christian P. R.

    2017-08-01

    Functional antibody delivery in living cells would enable the labelling and manipulation of intracellular antigens, which constitutes a long-thought goal in cell biology and medicine. Here we present a modular strategy to create functional cell-permeable nanobodies capable of targeted labelling and manipulation of intracellular antigens in living cells. The cell-permeable nanobodies are formed by the site-specific attachment of intracellularly stable (or cleavable) cyclic arginine-rich cell-penetrating peptides to camelid-derived single-chain VHH antibody fragments. We used this strategy for the non-endocytic delivery of two recombinant nanobodies into living cells, which enabled the relocalization of the polymerase clamp PCNA (proliferating cell nuclear antigen) and tumour suppressor p53 to the nucleolus, and thereby allowed the detection of protein-protein interactions that involve these two proteins in living cells. Furthermore, cell-permeable nanobodies permitted the co-transport of therapeutically relevant proteins, such as Mecp2, into the cells. This technology constitutes a major step in the labelling, delivery and targeted manipulation of intracellular antigens. Ultimately, this approach opens the door towards immunostaining in living cells and the expansion of immunotherapies to intracellular antigen targets.

  12. Optimizing laser-driven proton acceleration from overdense targets

    PubMed Central

    Stockem Novo, A.; Kaluza, M. C.; Fonseca, R. A.; Silva, L. O.

    2016-01-01

    We demonstrate how to tune the main ion acceleration mechanism in laser-plasma interactions to collisionless shock acceleration, thus achieving control over the final ion beam properties (e. g. maximum energy, divergence, number of accelerated ions). We investigate this technique with three-dimensional particle-in-cell simulations and illustrate a possible experimental realisation. The setup consists of an isolated solid density target, which is preheated by a first laser pulse to initiate target expansion, and a second one to trigger acceleration. The timing between the two laser pulses allows to access all ion acceleration regimes, ranging from target normal sheath acceleration, to hole boring and collisionless shock acceleration. We further demonstrate that the most energetic ions are produced by collisionless shock acceleration, if the target density is near-critical, ne ≈ 0.5 ncr. A scaling of the laser power shows that 100 MeV protons may be achieved in the PW range. PMID:27435449

  13. Behavior of yeast cells in aqueous suspension affected by pulsed electric field.

    PubMed

    El Zakhem, H; Lanoisellé, J-L; Lebovka, N I; Nonus, M; Vorobiev, E

    2006-08-15

    This work discusses pulsed electric fields (PEF) induced effects in treatment of aqueous suspensions of concentrated yeast cells (S. cerevisiae). The PEF treatment was done using pulses of near-rectangular shape, electric field strength was within E=2-5 kV/cm and the total time of treatment was t(PEF)=10(-4)-0.1 s. The concentration of aqueous yeast suspensions was in the interval of C(Y)=0-22 (wt%), where 1% concentration corresponds to the cellular density of 2x10(8) cells/mL. Triton X-100 was used for studying non-ionic surfactant additive effects. The electric current peak value I was measured during each pulse application, and from these data the electrical conductivity sigma was estimated. The PEF-induced damage results in increase of sigma with t(PEF) increasing and attains its saturation level sigma approximately sigma(max) at long time of PEF treatment. The value of sigma(max) reflects the efficiency of damage. The reduced efficiency of damage at suspension volume concentration higher than phi(Y) approximately 32 vol% is explained by the percolation phenomenon in the randomly packed suspension of near-spherical cells. The higher cytoplasmic ions leakage was observed in presence of surfactant. Experiments were carried out in the static and continuous flow treatment chambers in order to reveal the effects of mixing in PEF-treatment efficiency. A noticeable aggregation of the yeast cells was observed in the static flow chamber during the PEF treatment, while aggregation was not so pronounced in the continuous flow chamber. The nature of the enhanced aggregation under the PEF treatment was revealed by the zeta-potential measurements: these data demonstrate different zeta-potential signs for alive and dead cells. The effect of the electric field strength on the PEF-induced extraction of the intracellular components of S. cerevisiae is discussed.

  14. Induction of viral interference by IPNV-carrier cells on target cells: A cell co-culture study.

    PubMed

    Parreño, Ricardo; Torres, Susana; Almagro, Lucía; Belló-Pérez, Melissa; Estepa, Amparo; Perez, Luis

    2016-11-01

    IPNV is a salmonid birnavirus that possesses the ability to establish asymptomatic persistent infections in a number of valuable fish species. The presence of IPNV may interfere with subsequent infection by other viruses. In the present study we show that an IPNV-carrier cell line (EPC IPNV ) can induce an antiviral state in fresh EPC by co-cultivating both cell types in three different ways: a "droplet" culture system, a plastic chamber setup, and a transmembrane (Transwell ® ) system. All three cell co-culture methods were proven useful to study donor/target cell interaction. Naïve EPC cells grown in contact with EPC IPNV cells develop resistance to VHSV superinfection. The transmembrane system seems best suited to examine gene expression in donor and target cells separately. Our findings point to the conclusion that one or more soluble factors produced by the IPNV carrier culture induce the innate immune response within the target cells. This antiviral response is associated to the up-regulation of interferon (ifn) and mx gene expression in target EPC cells. To our knowledge this is the first article describing co-culture systems to study the interplay between virus-carrier cells and naive cells in fish. Copyright © 2016 The Author(s). Published by Elsevier Ltd.. All rights reserved.

  15. Changes in the emission properties of metallic targets upon exposure to repetitively pulsed laser radiation

    NASA Astrophysics Data System (ADS)

    Konov, V. I.; Pimenov, S. M.; Prokhorov, A. M.; Chapliev, N. I.

    1988-02-01

    A scanning electron microscope and a repetitively pulsed CO2 laser are used to reveal the relationships which govern the correlation of the transforming metal surface microrelief with the emission of charged particles and the surface luminescence upon exposure to multipulse laser focusing. It is shown that the effect of sorption and laser-stimulated desorption on the emission signals can manifest itself in different ways depending on the current oscillation mode in the target-vacuum chamber circuit.

  16. Selection of Phage Display Peptides Targeting Human Pluripotent Stem Cell-Derived Progenitor Cell Lines.

    PubMed

    Bignone, Paola A; Krupa, Rachel A; West, Michael D; Larocca, David

    2016-01-01

    The ability of human pluripotent stem cells (hPS) to both self-renew and differentiate into virtually any cell type makes them a promising source of cells for cell-based regenerative therapies. However, stem cell identity, purity, and scalability remain formidable challenges that need to be overcome for translation of pluripotent stem cell research into clinical applications. Directed differentiation from hPS cells is inefficient and residual contamination with pluripotent cells that have the potential to form tumors remains problematic. The derivation of scalable (self-renewing) embryonic progenitor stem cell lines offers a solution because they are well defined and clonally pure. Clonally pure progenitor stem cell lines also provide a means for identifying cell surface targeting reagents that are useful for identification, tracking, and repeated derivation of the corresponding progenitor stem cell types from additional hPS cell sources. Such stem cell targeting reagents can then be applied to the manufacture of genetically diverse banks of human embryonic progenitor cell lines for drug screening, disease modeling, and cell therapy. Here we present methods to identify human embryonic progenitor stem cell targeting peptides by selection of phage display libraries on clonal embryonic progenitor cell lines and demonstrate their use for targeting quantum dots (Qdots) for stem cell labeling.

  17. Enhancement of cardiomyogenesis in stem cells by low intensity pulsed ultrasound

    NASA Astrophysics Data System (ADS)

    Teo, Ailing; Morshedi, Amir; Wang, Jen-Chieh; Lim, Mayasari; Zhou, Yufeng

    2017-03-01

    Low intensity pulsed ultrasound (LIPUS) has been shown to enhance bone and cartilage regeneration from stem cells. Gene expression of angiotensin II type 1 (AT1) receptor can be increased in LIPUS-treated osteoblasts. The AT1 receptor is a known mechanoreceptor in cardiomyocytes. It suggests that LIPUS may enhance cardiomyogenesis via mechanotransduction by increasing AT1 expression. Murine embryonic stem cells (ESCs) were treated daily by 10-min 1MHz LIPUS at spatial-average temporal-peak acoustic intensities of 30 mW/cm2 and 300 mW/cm2 in both continuous and pulsed wave (20% duty cycle) for 10 days. Polymerase chain reaction (PCR), immunocytochemistry, and beating rate were used to evaluate the cardiac viability quantitatively. After the treatment of LIPUS, beating rate of contractile areas and cardiac gene expression, such as α- and β-myosin heavy chain, were improved. Furthermore, no deleterious effects to the development of cardiac proteins were observed. All results suggest that LIPUS stimulation has the capacity of enhancing cardiomyogenesis from embryonic stem cells. With the benefit and the ease in incorporating LIPUS into various culture platforms, LIPUS has the potential to produce cardiomyocytes for clinical use in the future.

  18. Simulations of bremsstrahlung emission in ultra-intense laser interactions with foil targets

    NASA Astrophysics Data System (ADS)

    Vyskočil, Jiří; Klimo, Ondřej; Weber, Stefan

    2018-05-01

    Bremsstrahlung emission from interactions of short ultra-intense laser pulses with solid foils is studied using particle-in-cell (PIC) simulations. A module for simulating bremsstrahlung has been implemented in the PIC loop to self-consistently account for the dynamics of the laser–plasma interaction, plasma expansion, and the emission of gamma ray photons. This module made it possible to study emission from thin targets, where refluxing of hot electrons plays an important role. It is shown that the angular distribution of the emitted photons exhibits a four-directional structure with the angle of emission decreasing with the increase of the width of the target. Additionally, a collimated forward flash consisting of high energy photons has been identified in thin targets. The conversion efficiency of the energy of the laser pulse to the energy of the gamma rays rises with both the driving pulse intensity, and the thickness of the target. The amount of gamma rays also increases with the atomic number of the target material, despite a lower absorption of the driving laser pulse. The angular spectrum of the emitted gamma rays is directly related to the increase of hot electron divergence during their refluxing and its measurement can be used in experiments to study this process.

  19. Intracellular Retention of ABL Kinase Inhibitors Determines Commitment to Apoptosis in CML Cells

    PubMed Central

    Dziadosz, Marek; Schnöder, Tina; Heidel, Florian; Schemionek, Mirle; Melo, Junia V.; Kindler, Thomas; Müller-Tidow, Carsten; Koschmieder, Steffen; Fischer, Thomas

    2012-01-01

    Clinical development of imatinib in CML established continuous target inhibition as a paradigm for successful tyrosine kinase inhibitor (TKI) therapy. However, recent reports suggested that transient potent target inhibition of BCR-ABL by high-dose TKI (HD-TKI) pulse-exposure is sufficient to irreversibly commit cells to apoptosis. Here, we report a novel mechanism of prolonged intracellular TKI activity upon HD-TKI pulse-exposure (imatinib, dasatinib) in BCR-ABL-positive cells. Comprehensive mechanistic exploration revealed dramatic intracellular accumulation of TKIs which closely correlated with induction of apoptosis. Cells were rescued from apoptosis upon HD-TKI pulse either by repetitive drug wash-out or by overexpression of ABC-family drug transporters. Inhibition of ABCB1 restored sensitivity to HD-TKI pulse-exposure. Thus, our data provide evidence that intracellular drug retention crucially determines biological activity of imatinib and dasatinib. These studies may refine our current thinking on critical requirements of TKI dose and duration of target inhibition for biological activity of TKIs. PMID:22815843

  20. Coherent synchrotron emission in transmission with double foil target

    NASA Astrophysics Data System (ADS)

    Xu, X. R.; Qiao, B.; Chang, H. X.; Zhang, Y. X.; Zhang, H.; Zhong, C. L.; Zhou, C. T.; Zhu, S. P.; He, X. T.

    2018-04-01

    Generation of intense single attosecond pulses from coherent synchrotron emission (CSE), in the transmitted direction of the laser-irradiated double foil targets, has been investigated theoretically and numerically. Unlike conventional CSE in the single foil target case, here the dense electron nanobunch is formed in the vacuum gap between two foils, which is composed of the electrons blown out from the first ultrathin foil. Owing to the existence of the vacuum gap, the electron nanobunch can be accelerated to more energy. In addition, more laser energy can penetrate through the nanobunch and get reflected from the second foil. These reflected lasers and electron nanobunches interact with each other and results in enhanced CSE and consequently, the generation of intense attosecond pulses. Particle-in-cell simulations show that a single attosecond pulse with duration of 18 {as}, photon energy > 0.16 {keV} and peak intensity of 1.7× {10}20 {{W}}/{cm}}2 can be obtained from the double-foil targets irradiated by a laser at intensity of 7.7× {10}21 {{W}}/{cm}}2.

  1. Development of pulsed processes for the manufacture of solar cells. Quarterly progress report No. 3, April--July 1978

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    Not Available

    1978-07-01

    Third quarter results under a program to develop ion implantation and specialized, associated processes necessary to achieve automated production of silicon solar cells are described. An ion implantation facility development for solar cell production is described, and a design for an automated production implanter is presented. Also, solar cell development efforts using combined ion implantation and pulsed energy techniques are discussed. Cell performance comparisons have also been made in which junctions and back surface fields were prepared by diffusion and ion implantation. A model is presented to explain the mechanism of ion implantation damage annealing using pulsed energy sources. Functionalmore » requirements have been determined for a pulsed electron beam processor for annealing ion implantation damage at a rate compatible with a 100 milliampere ion implanter. These rates result in a throughput of 100 megawatts of solar cell product per year.« less

  2. The Quest for Targets Executing MYC-Dependent Cell Transformation.

    PubMed

    Hartl, Markus

    2016-01-01

    MYC represents a transcription factor with oncogenic potential converting multiple cellular signals into a broad transcriptional response, thereby controlling the expression of numerous protein-coding and non-coding RNAs important for cell proliferation, metabolism, differentiation, and apoptosis. Constitutive activation of MYC leads to neoplastic cell transformation, and deregulated MYC alleles are frequently observed in many human cancer cell types. Multiple approaches have been performed to isolate genes differentially expressed in cells containing aberrantly activated MYC proteins leading to the identification of thousands of putative targets. Functional analyses of genes differentially expressed in MYC-transformed cells had revealed that so far more than 40 upregulated or downregulated MYC targets are actively involved in cell transformation or tumorigenesis. However, further systematic and selective approaches are required for determination of the known or yet unidentified targets responsible for processing the oncogenic MYC program. The search for critical targets in MYC-dependent tumor cells is exacerbated by the fact that during tumor development, cancer cells progressively evolve in a multistep process, thereby acquiring their characteristic features in an additive manner. Functional expression cloning, combinatorial gene expression, and appropriate in vivo tests could represent adequate tools for dissecting the complex scenario of MYC-specified cell transformation. In this context, the central goal is to identify a minimal set of targets that suffices to phenocopy oncogenic MYC. Recently developed genomic editing tools could be employed to confirm the requirement of crucial transformation-associated targets. Knowledge about essential MYC-regulated genes is beneficial to expedite the development of specific inhibitors to interfere with growth and viability of human tumor cells in which MYC is aberrantly activated. Approaches based on the principle of

  3. The Quest for Targets Executing MYC-Dependent Cell Transformation

    PubMed Central

    Hartl, Markus

    2016-01-01

    MYC represents a transcription factor with oncogenic potential converting multiple cellular signals into a broad transcriptional response, thereby controlling the expression of numerous protein-coding and non-coding RNAs important for cell proliferation, metabolism, differentiation, and apoptosis. Constitutive activation of MYC leads to neoplastic cell transformation, and deregulated MYC alleles are frequently observed in many human cancer cell types. Multiple approaches have been performed to isolate genes differentially expressed in cells containing aberrantly activated MYC proteins leading to the identification of thousands of putative targets. Functional analyses of genes differentially expressed in MYC-transformed cells had revealed that so far more than 40 upregulated or downregulated MYC targets are actively involved in cell transformation or tumorigenesis. However, further systematic and selective approaches are required for determination of the known or yet unidentified targets responsible for processing the oncogenic MYC program. The search for critical targets in MYC-dependent tumor cells is exacerbated by the fact that during tumor development, cancer cells progressively evolve in a multistep process, thereby acquiring their characteristic features in an additive manner. Functional expression cloning, combinatorial gene expression, and appropriate in vivo tests could represent adequate tools for dissecting the complex scenario of MYC-specified cell transformation. In this context, the central goal is to identify a minimal set of targets that suffices to phenocopy oncogenic MYC. Recently developed genomic editing tools could be employed to confirm the requirement of crucial transformation-associated targets. Knowledge about essential MYC-regulated genes is beneficial to expedite the development of specific inhibitors to interfere with growth and viability of human tumor cells in which MYC is aberrantly activated. Approaches based on the principle of

  4. Targeting human breast cancer cells by an oncolytic adenovirus using microRNA-targeting strategy.

    PubMed

    Shayestehpour, Mohammad; Moghim, Sharareh; Salimi, Vahid; Jalilvand, Somayeh; Yavarian, Jila; Romani, Bizhan; Mokhtari-Azad, Talat

    2017-08-15

    MicroRNA-targeting strategy is a promising approach that enables oncolytic viruses to replicate in tumor cells but not in normal cells. In this study, we targeted adenoviral replication toward breast cancer cells by inserting ten complementary binding sites for miR-145-5p downstream of E1A gene. In addition, we evaluated the effect of increasing miR-145 binding sites on inhibition of virus replication. Ad5-control and adenoviruses carrying five or ten copies of miR145-5p target sites (Ad5-5miR145T, Ad5-10miR145T) were generated and inoculated into MDA-MB-453, BT-20, MCF-7 breast cancer cell lines and human mammary epithelial cells (HMEpC). Titer of Ad5-10miR145T in HMEpC was significantly lower than Ad5-control titer. Difference between the titer of these two viruses at 12, 24, 36, and 48h after infection was 1.25, 2.96, 3.06, and 3.77 log TCID 50 . No significant difference was observed between the titer of both adenoviruses in MDA-MB-453, BT-20 and MCF-7 cells. The infectious titer of adenovirus containing 10 miR-145 binding sites in HMEpC cells at 24, 36, and 48h post-infection was 1.7, 2.08, and 4-fold, respectively, lower than the titer of adenovirus carrying 5 miR-145 targets. Our results suggest that miR-145-targeting strategy provides selectivity for adenovirus replication in breast cancer cells. Increasing the number of miRNA binding sites within the adenoviral genome confers more selectivity for viral replication in cancer cells. Copyright © 2017. Published by Elsevier B.V.

  5. Selective Gene Transfection of Individual Cells In Vitro with Plasmonic Nanobubbles

    PubMed Central

    Lukianova-Hleb, Ekaterina; Samaniego, Adam P.; Wen, Jianguo; Metelitsa, Leonid; Chang, Chung-Che; Lapotko, Dmitri

    2011-01-01

    Gene delivery and transfection of eukaryotic cells is widely used for research and for developing gene cell therapy. However, the existing methods lack selectivity, efficacy and safety when heterogeneous cell systems must be treated. We report a new method that employs plasmonic nanobubbles (PNBs) for delivery and transfection. A PNB is a novel, tunable cellular agent with a dual mechanical and optical action due to the formation of the vapor nanobubble around a transiently heated gold nanoparticle upon its exposure to a laser pulse. PNBs enabled the mechanical injection of the extracellular cDNA plasmid into the cytoplasm of individual target living cells, cultured leukemia cells and human CD34+CD117+ stem cells and expression of a green fluorescent protein (GFP) in those cells. PNB generation and lifetime correlated with the expression of green fluorescent protein in PNB-treated cells. Optical scattering by PNBs additionally provided the detection of the target cells and the guidance of cDNA injection at single cell level. In both cell models PNBs demonstrated a gene transfection effect in a single pulse treatment with high selectivity, efficacy and safety. Thus, PNBs provided targeted gene delivery at the single cell level in a single pulse procedure that can be used for safe and effective gene therapy. PMID:21315120

  6. Cell cycle-tailored targeting of metastatic melanoma: Challenges and opportunities.

    PubMed

    Haass, Nikolas K; Gabrielli, Brian

    2017-07-01

    The advent of targeted therapies of metastatic melanoma, such as MAPK pathway inhibitors and immune checkpoint antagonists, has turned dermato-oncology from the "bad guy" to the "poster child" in oncology. Current targeted therapies are effective, although here is a clear need to develop combination therapies to delay the onset of resistance. Many antimelanoma drugs impact on the cell cycle but are also dependent on certain cell cycle phases resulting in cell cycle phase-specific drug insensitivity. Here, we raise the question: Have combination trials been abandoned prematurely as ineffective possibly only because drug scheduling was not optimized? Firstly, if both drugs of a combination hit targets in the same melanoma cell, cell cycle-mediated drug insensitivity should be taken into account when planning combination therapies, timing of dosing schedules and choice of drug therapies in solid tumors. Secondly, if the combination is designed to target different tumor cell subpopulations of a heterogeneous tumor, one drug effective in a particular subpopulation should not negatively impact on the other drug targeting another subpopulation. In addition to the role of cell cycle stage and progression on standard chemotherapeutics and targeted drugs, we discuss the utilization of cell cycle checkpoint control defects to enhance chemotherapeutic responses or as targets themselves. We propose that cell cycle-tailored targeting of metastatic melanoma could further improve therapy outcomes and that our real-time cell cycle imaging 3D melanoma spheroid model could be utilized as a tool to measure and design drug scheduling approaches. © 2017 John Wiley & Sons A/S. Published by John Wiley & Sons Ltd.

  7. Experimental evidence for short-pulse laser heating of solid-density target to high bulk temperatures.

    PubMed

    Soloviev, A; Burdonov, K; Chen, S N; Eremeev, A; Korzhimanov, A; Pokrovskiy, G V; Pikuz, T A; Revet, G; Sladkov, A; Ginzburg, V; Khazanov, E; Kuzmin, A; Osmanov, R; Shaikin, I; Shaykin, A; Yakovlev, I; Pikuz, S; Starodubtsev, M; Fuchs, J

    2017-09-22

    Heating efficiently solid-density, or even compressed, matter has been a long-sought goal in order to allow investigation of the properties of such state of matter of interest for various domains, e.g. astrophysics. High-power lasers, pinches, and more recently Free-Electron-Lasers (FELs) have been used in this respect. Here we show that by using the high-power, high-contrast "PEARL" laser (Institute of Applied Physics-Russian Academy of Science, Nizhny Novgorod, Russia) delivering 7.5 J in a 60 fs laser pulse, such coupling can be efficiently obtained, resulting in heating of a slab of solid-density Al of 0.8 µm thickness at a temperature of 300 eV, and with minimal density gradients. The characterization of the target heating is achieved combining X-ray spectrometry and measurement of the protons accelerated from the Al slab. The measured heating conditions are consistent with a three-temperatures model that simulates resistive and collisional heating of the bulk induced by the hot electrons. Such effective laser energy deposition is achieved owing to the intrinsic high contrast of the laser which results from the Optical Parametric Chirped Pulse Amplification technology it is based on, allowing to attain high target temperatures in a very compact manner, e.g. in comparison with large-scale FEL facilities.

  8. Sonoporation of endothelial cells by vibrating targeted microbubbles.

    PubMed

    Kooiman, Klazina; Foppen-Harteveld, Miranda; van der Steen, Antonius F W; de Jong, Nico

    2011-08-25

    Molecular imaging using ultrasound makes use of targeted microbubbles. In this study we investigated whether these microbubbles could also be used to induce sonoporation in endothelial cells. Lipid-coated microbubbles were targeted to CD31 and insonified at 1 MHz at low peak negative acoustic pressures at six sequences of 10 cycle sine-wave bursts. Vibration of the targeted microbubbles was recorded with the Brandaris-128 high-speed camera (~13 million frames per second). In total, 31 cells were studied that all had one microbubble (1.2-4.2 micron in diameter) attached per cell. After insonification at 80 kPa, 30% of the cells (n=6) had taken up propidium iodide, while this was 20% (n=1) at 120 kPa and 83% (n=5) at 200 kPa. Irrespective of the peak negative acoustic pressure, uptake of propidium iodide was observed when the relative vibration amplitude of targeted microbubbles was greater than 0.5. No relationship was found between the position of the microbubble on the cell and induction of sonoporation. This study shows that targeted microbubbles can also be used to induce sonoporation, thus making it possible to combine molecular imaging and drug delivery. Copyright © 2011 Elsevier B.V. All rights reserved.

  9. Hundreds MeV monoenergetic proton bunch from interaction of 10{sup 20-21} W/cm{sup 2} circularly polarized laser pulse with tailored complex target

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    Zhang, Z. M.; Laser Fusion Research Center, CAEP, Mianyang 621900; He, X. T.

    A complex target (CT) configuration tailored for generating high quality proton bunch by circularly polarized laser pulses at intensities of 10{sup 20-21} W/cm{sup 2} is proposed. Two-dimensional particle-in-cell simulations show that both the collimation and mono-energetic qualities of the accelerated proton bunch obtained using a front-shaped thin foil can be greatly enhanced by the backside inhomogeneous plasma layer. The main mechanisms for improving the accelerated protons are identified and discussed. These include stabilization of the photon cavity, providing hole-boring supplementary acceleration and suppressing the thermal-electron effects. A theory for tailoring the CT parameters is also presented.

  10. Targeted silver nanoparticles for ratiometric cell phenotyping

    NASA Astrophysics Data System (ADS)

    Willmore, Anne-Mari A.; Simón-Gracia, Lorena; Toome, Kadri; Paiste, Päärn; Kotamraju, Venkata Ramana; Mölder, Tarmo; Sugahara, Kazuki N.; Ruoslahti, Erkki; Braun, Gary B.; Teesalu, Tambet

    2016-04-01

    Affinity targeting is used to deliver nanoparticles to cells and tissues. For efficient targeting, it is critical to consider the expression and accessibility of the relevant receptors in the target cells. Here, we describe isotopically barcoded silver nanoparticles (AgNPs) as a tool for auditing affinity ligand receptors in cells. Tumor penetrating peptide RPARPAR (receptor: NRP-1) and tumor homing peptide GKRK (receptor: p32) were used as affinity ligands on the AgNPs. The binding and uptake of the peptide-functionalized AgNPs by cultured PPC-1 prostate cancer and M21 melanoma cells was dependent on the cell surface expression of the cognate peptide receptors. Barcoded peptide-functionalized AgNPs were synthesized from silver and palladium isotopes. The cells were incubated with a cocktail of the barcoded nanoparticles [RPARPAR (R), GKRK (K), and control], and cellular binding and internalization of each type of nanoparticle was assessed by inductively coupled plasma mass spectrometry. The results of isotopic analysis were in agreement with data obtained using optical methods. Using ratiometric measurements, we were able to classify the PPC-1 cell line as mainly NRP-1-positive, with 75 +/- 5% R-AgNP uptake, and the M21 cell line as only p32-positive, with 89 +/- 9% K-AgNP uptake. The isotopically barcoded multiplexed AgNPs are useful as an in vitro ratiometric phenotyping tool and have potential uses in functional evaluation of the expression of accessible homing peptide receptors in vivo.Affinity targeting is used to deliver nanoparticles to cells and tissues. For efficient targeting, it is critical to consider the expression and accessibility of the relevant receptors in the target cells. Here, we describe isotopically barcoded silver nanoparticles (AgNPs) as a tool for auditing affinity ligand receptors in cells. Tumor penetrating peptide RPARPAR (receptor: NRP-1) and tumor homing peptide GKRK (receptor: p32) were used as affinity ligands on the AgNPs. The

  11. A novel double-targeted nondrug delivery system for targeting cancer stem cells

    PubMed Central

    Qiao, Shupei; Zhao, Yufang; Geng, Shuai; Li, Yong; Hou, Xiaolu; Liu, Yi; Lin, Feng-Huei; Yao, Lifen; Tian, Weiming

    2016-01-01

    Instead of killing cancer stem cells (CSCs), the conventional chemotherapy used for cancer treatment promotes the enrichment of CSCs, which are responsible for tumor growth, metastasis, and recurrence. However, most therapeutic agents are only able to kill a small proportion of CSCs by targeting one or two cell surface markers or dysregulated CSC pathways, which are usually shared with normal stem cells (NSCs). In this study, we developed a novel nondrug delivery system for the dual targeting of CSCs by conjugating hyaluronic acid (HA) and grafting the doublecortin-like kinase 1 (DCLK1) monoclonal antibody to the surface of poly(ethylene glycol) (PEG)–poly(d,l-lactide-co-glycolide) (PLGA) nanoparticles (NPs), which can specifically target CD44 receptors and the DCLK1 surface marker – the latter was shown to possess the capacity to distinguish between CSCSs and NSCs. The size and morphology of these NPs were characterized by dynamic light scattering (DLS), transmission electron microscopy (TEM), and scanning electron microscopy (SEM). This was followed by studies of NP encapsulation efficiency and in vitro drug release properties. Then, the cytotoxicity of the NPs was tested via Cell Counting Kit-8 assay. Finally, the 4T1 CSCs were obtained from the alginate-based platform, which we developed as an in vitro tumor model. Tumor-bearing nude mice were used as in vivo models to systematically detect the ability of NPs to target CSCs. Our results showed that the DCLK1–HA–PEG–PLGA NPs exhibited a targeting effect toward CSCs both in vitro and in vivo. These findings have important implications for the rational design of drug delivery systems that target CSCs with high efficacy. PMID:27994463

  12. Efficient optical pulse stacker system

    DOEpatents

    Seppala, Lynn G.; Haas, Roger A.

    1982-01-01

    Method and apparatus for spreading and angle-encoding each pulse of a multiplicity of small area, short pulses into several temporally staggered pulses by use of appropriate beam splitters, with the optical elements being arranged so that each staggered pulse is contiguous with one or two other such pulses, and the entire sequence of stacked pulses comprising a single, continuous long pulse. The single long pulse is expanded in area, and then doubly passed through a nonstorage laser amplifier such as KrF. After amplification, the physically separated, angle-encoded and temporally staggered pulses are recombined into a single pulse of short duration. This high intensity output beam is well collimated and may be propagated over long distance, or used for irradiating inertial confinement fusion targets.

  13. Advanced cell therapies: targeting, tracking and actuation of cells with magnetic particles.

    PubMed

    Connell, John J; Patrick, P Stephen; Yu, Yichao; Lythgoe, Mark F; Kalber, Tammy L

    2015-01-01

    Regenerative medicine would greatly benefit from a new platform technology that enabled measurable, controllable and targeting of stem cells to a site of disease or injury in the body. Superparamagnetic iron-oxide nanoparticles offer attractive possibilities in biomedicine and can be incorporated into cells, affording a safe and reliable means of tagging. This review describes three current and emerging methods to enhance regenerative medicine using magnetic particles to guide therapeutic cells to a target organ; track the cells using MRI and assess their spatial localization with high precision and influence the behavior of the cell using magnetic actuation. This approach is complementary to the systemic injection of cell therapies, thus expanding the horizon of stem cell therapeutics.

  14. Phototransfection of mouse embryonic stem cells with plasmid DNA using femtosecond laser pulses

    NASA Astrophysics Data System (ADS)

    Thobakgale, Lebogang; Manoto, Sello Lebohang; Ombinda Lemboumba, Saturnin; Maaza, Malik; Mthunzi-Kufa, Patience

    2017-02-01

    Cellular manipulation by delivery of molecules into cells has been applied extensively in tissue engineering research for medical applications . The different molecular delivery techniques used range from viral and chemical agents to physical and electrical methods. Although successful in most studies, these techniques have inherent difficulties such as toxicity, unwanted genetic mutations and low reproducibility respectively. Literature recognizes pulsed lasers at femtosecond level to be most efficient in photonic interactions with biological material. As of late, laser pulses have been used for drug and DNA delivery into cells via transient optical perforation of the cellular membrane. Thus in this study, we design and construct an optical system coupled to a femtosecond laser for the purpose of phototransfection or insertion of plasmid DNA (pDNA) into cells using lasers. We used fluorescent green protein (pGFP) to transfect mouse embryonic stem cells as our model. Secondly, we applied fluorescence imaging to view the extent of DNA delivery using this method. We also assessed the biocompatibility of our system by performing molecular assays of the cells post irradiation using adenosine triphosphate (ATP) and lactate dehydrogenase (LDH).

  15. A double-pulse approach for electrotransfection.

    PubMed

    Pasquet, L; Bellard, E; Golzio, M; Rols, M P; Teissie, J

    2014-12-01

    Gene transfer and expression can be obtained by delivering calibrated electric pulses on cells in the presence of plasmids coding for the activity of interest. The electric treatment affects the plasma membrane and induces the formation of a transient complex between nucleic acids and the plasma membrane. It results in a delivery of the plasmid in the cytoplasm. Expression is only obtained if the plasmid is translocated inside the nucleus. This is a key limit in the process. We previously showed that delivery of a high-field short-duration electric pulse was inducing a structural alteration of the nuclear envelope. This study investigates if the double-pulse approach (first pulse to transfer the plasmid to the cytoplasm, and second pulse to induce the structural alteration of the envelope) was a way to enhance the protein expression using the green fluorescent protein as a reporter. We observed that not only the double-pulse approach induced the transfection of a lower number of cells but moreover, these transfected cells were less fluorescent than the cells treated only with the first pulse.

  16. Self-targeted salinomycin-loaded DSPE-PEG-methotrexate nanomicelles for targeting both head and neck squamous cell carcinoma cancer cells and cancer stem cells.

    PubMed

    Zhu, Minhui; Chen, Shicai; Hua, Libo; Zhang, Caiyun; Chen, Mengjie; Chen, Donghui; Dong, Yinmei; Zhang, Yingying; Li, Meng; Song, Xianmin; Chen, Huaiwen; Zheng, Hongliang

    2017-02-01

    To target both head and neck squamous cell carcinoma (HNSCC) cells and cancer stem cells (CSCs) by salinomycin-loaded DSPE-PEG-MTX (synthesized using DSPE-PEG2000-NH2 and methotrexate) nanomicelles (M-SAL-MTX). The characterization, antitumor activity and mechanism of M-SAL-MTX were evaluated. M-SAL-MTX showed enhanced inhibitory effect toward both HNSCC CSCs and non-CSCs compared with a single treatment of methotrexate and salinomycin. In nude mice-bearing HNSCC xenografts, M-SAL-MTX suppressed tumor growth more effectively than other controls including combination of methotrexate and salinomycin. Therefore, M-SAL-MTX may provide a strategy for treating HNSCC by targeting both HNSCC CSCs and HNSCC cells.

  17. Photothermal therapy to damage PC3 cancer cells: in vitro studies of a pulsed laser (Conference Presentation)

    NASA Astrophysics Data System (ADS)

    Zamora-Romero, Noe; Aguilar, Guillermo; Devia-Cruz, Luis F.; Banks, Darren; Zhang, Bin; Halaney, David L.

    2017-02-01

    Laser-nanoparticles interactions have been widely used for several years. In biomedicine, several in vitro and in vivo experiments have shown promising results for the detection and treatment of cancer. One of the techniques of interest to us, is the nanoparticle-assisted photothermal therapy (PTT), which consists of irradiating cancer cells incubated with nanoparticles with either a pulsed or continuous (cw) laser in order to damage the cells. However, there is still a debate over which type of laser is most effective for PTT for cancer treatment. On the one hand, cw lasers are minimally invasive and can be used for both detection and treatment of tumors. On the other hand, pulsed lasers offer great spatial precision and can deposit higher energy fluences than cw lasers, making them very efficient for inducing cavitation to damage cancer cells and tumors mechanically. The aim of this study is to investigate whether simultaneous application of cw and pulsed laser could offer a synergetic enhancement of PTT efficacy to damage cancer cells in vitro, compared to either laser applied individually. PTT efficacy is evaluated through cell viability tests following the irradiation of prostate cancer (PC3) cells incubated with gold nanorods (5.7 X10 10 p/ml). By irradiating the PC3-nanorod solution with the cw laser at 808 nm for 60 seconds, the temperature increases from 37.5 to 45°C, which damages some cancer cells via the heat shock response within the cells, and also could increase their sensitivity to the mechanical stress caused by the shock wave generated from inducing cavitation in the solution by the pulsed laser irradiation.

  18. Nail-like targets for laser plasma interaction experiments

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    Pasley, J; Wei, M; Shipton, E

    2007-12-18

    The interaction of ultra-high power picosecond laser pulses with solid targets is of interest both for benchmarking the results of hybrid particle in cell (PIC) codes and also for applications to re-entrant cone guided fast ignition. We describe the construction of novel targets in which copper/titanium wires are formed into 'nail-like' objects by a process of melting and micromachining, so that energy can be reliably coupled to a 24 {micro}m diameter wire. An extreme-ultraviolet image of the interaction of the Titan laser with such a target is shown.

  19. An innovative pre-targeting strategy for tumor cell specific imaging and therapy

    NASA Astrophysics Data System (ADS)

    Qin, Si-Yong; Peng, Meng-Yun; Rong, Lei; Jia, Hui-Zhen; Chen, Si; Cheng, Si-Xue; Feng, Jun; Zhang, Xian-Zheng

    2015-08-01

    A programmed pre-targeting system for tumor cell imaging and targeting therapy was established based on the ``biotin-avidin'' interaction. In this programmed functional system, transferrin-biotin can be actively captured by tumor cells with the overexpression of transferrin receptors, thus achieving the pre-targeting modality. Depending upon avidin-biotin recognition, the attachment of multivalent FITC-avidin to biotinylated tumor cells not only offered the rapid fluorescence labelling, but also endowed the pre-targeted cells with targeting sites for the specifically designed biotinylated peptide nano-drug. Owing to the successful pre-targeting, tumorous HepG2 and HeLa cells were effectively distinguished from the normal 3T3 cells via fluorescence imaging. In addition, the self-assembled peptide nano-drug resulted in enhanced cell apoptosis in the observed HepG2 cells. The tumor cell specific pre-targeting strategy is applicable for a variety of different imaging and therapeutic agents for tumor treatments.A programmed pre-targeting system for tumor cell imaging and targeting therapy was established based on the ``biotin-avidin'' interaction. In this programmed functional system, transferrin-biotin can be actively captured by tumor cells with the overexpression of transferrin receptors, thus achieving the pre-targeting modality. Depending upon avidin-biotin recognition, the attachment of multivalent FITC-avidin to biotinylated tumor cells not only offered the rapid fluorescence labelling, but also endowed the pre-targeted cells with targeting sites for the specifically designed biotinylated peptide nano-drug. Owing to the successful pre-targeting, tumorous HepG2 and HeLa cells were effectively distinguished from the normal 3T3 cells via fluorescence imaging. In addition, the self-assembled peptide nano-drug resulted in enhanced cell apoptosis in the observed HepG2 cells. The tumor cell specific pre-targeting strategy is applicable for a variety of different imaging

  20. Stromal cells in breast cancer as a potential therapeutic target

    PubMed Central

    Dykes, Samantha S.; Hughes, Veronica S.; Wiggins, Jennifer M.; Fasanya, Henrietta O.; Tanaka, Mai; Siemann, Dietmar

    2018-01-01

    Breast cancer in the United States is the second most commonly diagnosed cancer in women. About 1 in 8 women will develop invasive breast cancer over the course of her lifetime and breast cancer remains the second leading cause of cancer-related death. In pursuit of novel therapeutic strategies, researchers have examined the tumor microenvironment as a potential anti-cancer target. In addition to neoplastic cells, the tumor microenvironment is composed of several critical normal cell types, including fibroblasts, vascular and lymph endothelial cells, osteoclasts, adipocytes, and immune cells. These cells have important roles in healthy tissue stasis, which frequently are altered in tumors. Indeed, tumor-associated stromal cells often contribute to tumorigenesis, tumor progression, and metastasis. Consequently, these host cells may serve as a possible target in anti-tumor and anti-metastatic therapeutic strategies. Targeting the tumor associated host cells offers the benefit that such cells do not mutate and develop resistance in response to treatment, a major cause of failure in cancer therapeutics targeting neoplastic cells. This review discusses the role of host cells in the tumor microenvironment during tumorigenesis, progression, and metastasis, and provides an overview of recent developments in targeting these cell populations to enhance cancer therapy efficacy.

  1. Oxygen Saturation Targeting by Pulse Oximetry (SpO2) in the Extremely Low Gestational Age Neonate (ELGAN): A Quixotic Quest

    PubMed Central

    Cummings, James J.; Lakshminrusimha, Satyan

    2017-01-01

    Purpose of review A collaboration of comparative effectiveness research trials of pulse oximeter saturation (SpO2) targeting in extremely low gestational age neonates (ELGANs) have begun to report their aggregate results. We will examine the results of those trials, collectively referred to as the Neonatal Oxygenation Prospective Meta-analysis, or NeOProM. We will also discuss the uncertainties that remain and the clinical challenges that lie ahead. Recent findings The primary outcome from NeOProM was a composite of death or disability at 18–24 months corrected age. Earlier this year, the last of these reports was published. Although there were no differences in the primary outcome overall, analyses of secondary outcomes and data subsets following a pulse oximeter revision show significant treatment differences between targeting a lower compared to a higher SpO2. Summary NeOProM represents the largest collaborative clinical research study of SpO2 targets in ELGANs. While aggregate results give us some insight into the feasibility and efficacy of SpO2 targeting in this population, many questions remain. A patient-level analysis, tracking individual outcomes based on actual SpO2 experienced, may shed some light on these questions. However, finding a single optimal SpO2 range seems unlikely. PMID:28085683

  2. Manipulation of cell membrane using carbon nanotube scaffold as a photoresponsive stimuli generator.

    PubMed

    Sada, Takao; Fujigaya, Tsuyohiko; Nakashima, Naotoshi

    2014-08-01

    We describe, for the first time, the perforation of the cell membrane in the targeted single cell based on the nanosecond pulsed near-infrared (NIR) laser irradiation of a thin film of carbon nanotubes that act as an effective photon absorber as well as stimuli generator. When the power of NIR laser is over 17.5 μ J/pulse, the cell membrane after irradiation is irreversibly disrupted and results in cell death. In sharp contrast, the perforation of the cell membrane occurs at suitable laser power (∼15 μ J/pulse) without involving cell termination.

  3. Intense Non-Linear Soft X-Ray Emission from a Hydride Target during Pulsed D Bombardment

    NASA Astrophysics Data System (ADS)

    Miley, George H.; Yang, Yang; Lipson, Andrei; Haque, Munima; Percel, Ian; Romer, Michael

    Radiation emission from low-energy nuclear radiation (LENR) electrodes (both charged-particle and X-rays) represents an important feature of LENR in general. Here, calibration, measurement techniques, and soft X-ray emission results from deuterium bombardment of a Pd target (cathode) placed in a pulsed deuterium glow discharge (PGD) are described. An X-ray intensity of 13.4 mW/cm2 and a dose of 3.3 μJ/cm2 were calculated over a 0.5 ms pulse time from AXUV photodiode radiation detector measurements. A most striking feature is that X-ray energies >600 V are observed with a discharge voltage only about half of that value. To further investigate this phenomenon, emission during room temperature D-desorption from electrolytically loaded Pd:Dx cathodes was also studied. The X-ray emission energy observed was quite similar to the PGD case. However, the intensity in this case was almost 13 orders of magnitude lower due to the much lower deuterium fluxes involved.

  4. Experimental characterization and numerical modeling of tissue electrical conductivity during pulsed electric fields for irreversible electroporation treatment planning.

    PubMed

    Neal, Robert E; Garcia, Paulo A; Robertson, John L; Davalos, Rafael V

    2012-04-01

    Irreversible electroporation is a new technique to kill cells in targeted tissue, such as tumors, through a nonthermal mechanism using electric pulses to irrecoverably disrupt the cell membrane. Treatment effects relate to the tissue electric field distribution, which can be predicted with numerical modeling for therapy planning. Pulse effects will change the cell and tissue properties through thermal and electroporation (EP)-based processes. This investigation characterizes these changes by measuring the electrical conductivity and temperature of ex vivo renal porcine tissue within a single pulse and for a 200 pulse protocol. These changes are incorporated into an equivalent circuit model for cells and tissue with a variable EP-based resistance, providing a potential method to estimate conductivity as a function of electric field and pulse length for other tissues. Finally, a numerical model using a human kidney volumetric mesh evaluated how treatment predictions vary when EP- and temperature-based electrical conductivity changes are incorporated. We conclude that significant changes in predicted outcomes will occur when the experimental results are applied to the numerical model, where the direction and degree of change varies with the electric field considered.

  5. Mast cell proteases as pharmacological targets

    PubMed Central

    Caughey, George H.

    2015-01-01

    Mast cells are rich in proteases, which are the major proteins of intracellular granules and are released with histamine and heparin by activated cells. Most of these proteases are active in the granule as well outside of the mast cell when secreted, and can cleave targets near degranulating mast cells and in adjoining tissue compartments. Some proteases released from mast cells reach the bloodstream and may have far-reaching actions. In terms of relative amounts, the major mast cell proteases include the tryptases, chymases, cathepsin G, carboxypeptidase A3, dipeptidylpeptidase I/cathepsin C, and cathepsins L and S. Some mast cells also produce granzyme B, plasminogen activators, and matrix metalloproteinases. Tryptases and chymases are almost entirely mast cell-specific, whereas other proteases, such as cathepsins G, C, and L are expressed by a variety of inflammatory cells. Carboxypeptidase A3 expression is a property shared by basophils and mast cells. Other proteases, such as mastins, are largely basophil-specific, although human basophils are protease-deficient compared with their murine counterparts. The major classes of mast cell proteases have been targeted for development of therapeutic inhibitors. Also, a human β-tryptase has been proposed as a potential drug itself, to inactivate of snake venins. Diseases linked to mast cell proteases include allergic diseases, such as asthma, eczema, and anaphylaxis, but also include non-allergic diseases such inflammatory bowel disease, autoimmune arthritis, atherosclerosis, aortic aneurysms, hypertension, myocardial infarction, heart failure, pulmonary hypertension and scarring diseases of lungs and other organs. In some cases, studies performed in mouse models suggest protective or homeostatic roles for specific proteases (or groups of proteases) in infections by bacteria, worms and other parasites, and even in allergic inflammation. At the same time, a clearer picture has emerged of differences in the properties

  6. Efficient neutron production from sub-nanosecond laser pulse accelerating deuterons on target front side

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    Klir, D.; Institute of Plasma Physics, ASCR, Za Slovankou 3, 182 00 Prague 8; Institute of Physics, ASCR, Na Slovance 2, 182 21 Prague 8

    2015-09-15

    Neutron-producing experiments have been carried out on the Prague Asterix Laser System. At the fundamental wavelength of 1.315 μm, the laser pulse of a 600 J energy and 300 ps duration was focused on a thick deuterated-polyethylene target. Neutron yields reached (4.1 ± 0.8) × 10{sup 8} at the peak intensity of ≈3 × 10{sup 16 }W/cm{sup 2}. A more detailed analysis of neutron time-of-flight signals showed that a significant fraction of neutron yields was produced both by the {sup 2}H(d,n){sup 3}He reaction and by other neutron-producing reactions. Neutron energies together with delayed neutron and gamma emission showed that MeV deuterons escaped from a laser-produced plasma and interacted ≈50 nsmore » later with a borosilicate blast-shield glass. In order to increase DD neutron yields and to characterize deuteron beams via nuclear reactions, a secondary deuterated polyethylene target was used in a pitcher-catcher scheme at the target front side. In this experimental arrangement, the neutron yield reached (2.0 ± 0.5) × 10{sup 9} with the peak neutron fluence of (2.5 ± 0.5) × 10{sup 8 }n/sr. From the neutron yield, it was calculated that the secondary target was bombarded by 2 × 10{sup 14} deuterons in the 0.5–2.0 MeV energy range. The neutron yield of 2 × 10{sup 9} at the laser energy of 600 J implied the production efficiency of 3 × 10{sup 6 }n/J. A very important result is that the efficient neutron production was achieved with the low contrast, sub-nanosecond laser pulse of the intensity of 10{sup 16 }W/cm{sup 2}. The latter parameters can be achieved in a rep-rate mode more easily than ultra-high intensities and contrasts.« less

  7. PULSE AMPLITUDE ANALYZER

    DOEpatents

    Gray, G.W.; Jensen, A.S.

    1957-10-22

    A pulse-height analyzer system of improved design for sorting and counting a series of pulses, such as provided by a scintillation detector in nuclear radiation measurements, is described. The analyzer comprises a main transmission line, a cathode-ray tube for each section of the line with its deflection plates acting as the line capacitance; means to bias the respective cathode ray tubes so that the beam strikes a target only when a prearranged pulse amplitude is applied, with each tube progressively biased to respond to smaller amplitudes; pulse generating and counting means associated with each tube to respond when the beam is deflected; a control transmission line having the same time constant as the first line per section with pulse generating means for each tube for initiating a pulse on the second transmission line when a pulse triggers the tube of corresponding amplitude response, the former pulse acting to prevent successive tubes from responding to the pulse under test. This arrangement permits greater deflection sensitivity in the cathode ray tube and overcomes many of the disadvantages of prior art pulse-height analyzer circuits.

  8. Targeting Stromal Recruitment by Prostate Cancer Cells

    DTIC Science & Technology

    2006-03-01

    Ensinger, C., Tumer , Z., Tommerup, N. et al.: Hedgehog signaling in small-cell lung cancer : frequent in vivo but a rare event in vitro. Lung Cancer , 52...W81XWH-04-1-0157 TITLE: Targeting Stromal Recruitment by Prostate Cancer Cells PRINCIPAL INVESTIGATOR: Jingxian Zhang, Ph.D...DATES COVERED (From - To) 15 Feb 2004 – 14 Feb 2006 4. TITLE AND SUBTITLE 5a. CONTRACT NUMBER Targeting Stromal Recruitment by Prostate Cancer

  9. Ultrasonic Analysis of Peptide- and Antibody-Targeted Microbubble Contrast Agents for Molecular Imaging of αvβ3-Expressing Cells

    PubMed Central

    Dayton, Paul A.; Pearson, David; Clark, Jarrod; Simon, Scott; Schumann, Patricia A.; Zutshi, Reena; Matsunaga, Terry O.; Ferrara, Katherine W.

    2008-01-01

    The goal of targeted ultrasound contrast agents is to significantly and selectively enhance the detection of a targeted vascular site. In this manuscript, three distinct contrast agents targeted to the αvβ3 integrin are examined. The αvβ3 integrin has been shown to be highly expressed on metastatic tumors and endothelial cells during neovascularization, and its expression has been shown to correlate with tumor grade. Specific adhesion of these contrast agents to αvβ3-expressing cell monolayers is demonstrated in vitro, and compared with that of nontargeted agents. Acoustic studies illustrate a backscatter amplitude increase from monolayers exposed to the targeted contrast agents of up to 13-fold (22 dB) relative to enhancement due to control bubbles. A linear dependence between the echo amplitude and bubble concentration was observed for bound agents. The decorrelation of the echo from adherent targeted agents is observed over successive pulses as a function of acoustic pressure and bubble density. Frequency–domain analysis demonstrates that adherent targeted bubbles exhibit high-amplitude narrowband echo components, in contrast to the primarily wideband response from free microbubbles. Results suggest that adherent targeted contrast agents are differentiable from free-floating microbubbles, that targeted contrast agents provide higher sensitivity in the detection of angiogenesis, and that conventional ultrasound imaging techniques such as signal subtraction or decorrelation detection can be used to detect integrin-expressing vasculature with sufficient signal-to-noise. PMID:15296677

  10. Widely-duration-tunable nanosecond pulse Nd:YVO4 laser based on double Pockels cells

    NASA Astrophysics Data System (ADS)

    He, Li-Jiao; Liu, Ke; Bo, Yong; Wang, Xiao-Jun; Yang, Jing; Liu, Zhao; Zong, Qing-Shuang; Peng, Qin-Jun; Cui, Da-Fu; Xu, Zu-Yan

    2018-05-01

    The development of duration-tunable pulse lasers with constant output power is important for scientific research and materials processing. We present a widely-duration-tunable nanosecond (ns) pulse Nd:YVO4 laser based on double Pockels cells (PCs), i.e. inserting an extra PC into a conventional electro-optic Q-switched cavity dumped laser resonator. Under the absorbed pump power of 24.9 W, the pulse duration is adjustable from 31.9 ns to 5.9 ns by changing the amplitude of the high voltage on the inserted PC from 1100 V to 4400 V at the pulse repetition rate of 10 kHz. The corresponding average output power is almost entirely maintained in the range of 3.5–4.1 W. This represents more than three times increase in pulse duration tunable regime and average power compared to previously reported results for duration-tunable ns lasers. The laser beam quality factor was measured to be M 2  <  1.18.

  11. Magnetic stem cell targeting to the inner ear

    NASA Astrophysics Data System (ADS)

    Le, T. N.; Straatman, L.; Yanai, A.; Rahmanian, R.; Garnis, C.; Häfeli, U. O.; Poblete, T.; Westerberg, B. D.; Gregory-Evans, K.

    2017-12-01

    Severe sensorineural deafness is often accompanied by a loss of auditory neurons in addition to injury of the cochlear epithelium and hair cell loss. Cochlear implant function however depends on a healthy complement of neurons and their preservation is vital in achieving optimal results. We have developed a technique to target mesenchymal stem cells (MSCs) to a deafened rat cochlea. We then assessed the neuroprotective effect of systematically delivered MSCs on the survival and function of spiral ganglion neurons (SGNs). MSCs were labeled with superparamagnetic nanoparticles, injected via the systemic circulation, and targeted using a magnetized cochlea implant and external magnet. Neurotrophic factor concentrations, survival of SGNs, and auditory function were assessed at 1 week and 4 weeks after treatments and compared against multiple control groups. Significant numbers of magnetically targeted MSCs (>30 MSCs/section) were present in the cochlea with accompanied elevation of brain-derived neurotrophic factor and glial cell-derived neurotrophic factor levels (p < 0.001). In addition we saw improved survival of SGNs (approximately 80% survival at 4 weeks). Hearing threshold levels in magnetically targeted rats were found to be significantly better than those of control rats (p < 0.05). These results indicate that magnetic targeting of MSCs to the cochlea can be accomplished with a magnetized cochlear permalloy implant and an external magnet. The targeted stem cells release neurotrophic factors which results in improved SGN survival and hearing recovery. Combining magnetic cell-based therapy and cochlear implantation may improve cochlear implant function in treating deafness.

  12. [Apoptosis of human lung carcinoma cell line GLC-82 induced by high power electromagnetic pulse].

    PubMed

    Cao, Xiao-zhe; Zhao, Mei-lan; Wang, De-wen; Dong, Bo

    2002-09-01

    Electromagnetic pulse (EMP) could be used for sterilization of food and the efficiency is higher than 2450 MHz continuous microwave done. This study was designed to evaluate the effect of electromagnetic pulse (EMP) on apoptosis of human lung carcinoma cell line GLC-82, so that to explore and develop therapeutic means for cancer. The injury changes in GLC-82 cells after irradiated with EMP (electric field intensity was 60 kV/m, 5 pulses/2 min) were analyzed by cytometry, MTT chronometry, and flow cytometry. The immunohistochemical SP staining was used to determine the expressions of bcl-2 protein and p53 protein. The stained positive cells were analyzed by CMIAS-II image analysis system at a magnification 400. All data were analyzed by SPSS8.0 software. EMP could obviously inhibited proliferation and activity of lung carcinoma cell line GLC-82. The absorbance value (A570) of MTT decreased immediately, at 0 h, 1 h, and 6 h after the GLC-82 cells irradiated by EMP as compared with control group. The highest apoptosis rate was found to reach 13.38% by flow cytometry at 6 h after EMP irradiation. Down-regulation of bcl-2 expression and up-regulation of p53 expression were induced by EMP. EMP promotes apoptosis of GLC-82 cells. At same time, EMP can down-regulate bcl-2 expression and up-regulate p53 expression in GLC-82 cells. The bcl-2 and the p53 protein may involve the apoptotic process.

  13. Surface-modified gold nanorods for specific cell targeting

    NASA Astrophysics Data System (ADS)

    Wang, Chan-Ung; Arai, Yoshie; Kim, Insun; Jang, Wonhee; Lee, Seonghyun; Hafner, Jason H.; Jeoung, Eunhee; Jung, Deokho; Kwon, Youngeun

    2012-05-01

    Gold nanoparticles (GNPs) have unique properties that make them highly attractive materials for developing functional reagents for various biomedical applications including photothermal therapy, targeted drug delivery, and molecular imaging. For in vivo applications, GNPs need to be prepared with very little or negligible cytotoxicitiy. Most GNPs are, however, prepared using growth-directing surfactants such as cetyl trimethylammonium bromide (CTAB), which are known to have considerable cytotoxicity. In this paper, we describe an approach to remove CTAB to a non-toxic concentration. We optimized the conditions for surface modification with methoxypolyethylene glycol thiol (mPEG), which replaced CTAB and formed a protective layer on the surface of gold nanorods (GNRs). The cytotoxicities of pristine and surface-modified GNRs were measured in primary human umbilical vein endothelial cells and human cell lines derived from hepatic carcinoma cells, embryonic kidney cells, and thyroid papillary carcinoma cells. Cytotoxicity assays revealed that treating cells with GNRs did not significantly affect cell viability except for thyroid papillary carcinoma cells. Thyroid cancer cells were more susceptible to residual CTAB, so CTAB had to be further removed by dialysis in order to use GNRs for thyroid cell targeting. PEGylated GNRs are further modified to present monoclonal antibodies that recognize a specific surface marker, Na-I symporter, for thyroid cells. Antibody-conjugated GNRs specifically targeted human thyroid cells in vitro.

  14. Chimeric antigen receptor T cells targeting Fc μ receptor selectively eliminate CLL cells while sparing healthy B cells.

    PubMed

    Faitschuk, Elena; Hombach, Andreas A; Frenzel, Lukas P; Wendtner, Clemens-Martin; Abken, Hinrich

    2016-09-29

    Adoptive cell therapy of chronic lymphocytic leukemia (CLL) with chimeric antigen receptor (CAR)-modified T cells targeting CD19 induced lasting remission of this refractory disease in a number of patients. However, the treatment is associated with prolonged "on-target off-tumor" toxicities due to the targeted elimination of healthy B cells demanding more selectivity in targeting CLL cells. We identified the immunoglobulin M Fc receptor (FcμR), also known as the Fas apoptotic inhibitory molecule-3 or TOSO, as a target for a more selective treatment of CLL by CAR T cells. FcμR is highly and consistently expressed by CLL cells; only minor levels are detected on healthy B cells or other hematopoietic cells. T cells with a CAR specific for FcμR efficiently responded toward CLL cells, released a panel of proinflammatory cytokines and lytic factors, like soluble FasL and granzyme B, and eliminated the leukemic cells. In contrast to CD19 CAR T cells, anti-FcμR CAR T cells did not attack healthy B cells. T cells with anti-FcμR CAR delayed outgrowth of Mec-1-induced leukemia in a xenograft mouse model. T cells from CLL patients in various stages of the disease, modified by the anti-FcμR CAR, purged their autologous CLL cells in vitro without reducing the number of healthy B cells, which is the case with anti-CD19 CAR T cells. Compared with the currently used therapies, the data strongly imply a superior therapeutic index of anti-FcμR CAR T cells for the treatment of CLL. © 2016 by The American Society of Hematology.

  15. High pulse rate high resolution optical radar system

    NASA Technical Reports Server (NTRS)

    Goss, W. C.; Burns, R. H.; Chi, K. (Inventor)

    1973-01-01

    The system is composed of an optical cavity with a laser and a mode locking means to build up an optical pulse. An optical switch is also provided within the cavity to convert the polarization of the optical pulse generated within the cavity. The optical switch comprises an electro-optical crystal driven by a time delayed driver circuit which is triggered by a coincident signal made from an optical pulse signal and a gating pulse signal. The converted optical pulse strikes a polarization sensitive prism and is deflected out of the cavity toward the pending target in the form of a pulse containing most of the optical energy generated by the laser in the pulse build-up period. After striking the target, the reflected energy is picked up by a transceiver with the total travel time of the pulse being recorded.

  16. Pressure waves in liquid mercury target from pulsed heat loads and the possible way controlling their effects

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    Ni, L.; Skala, K.

    1996-06-01

    In ESS project liquid metals are selected as the main target for the pulsed spallation neutron source. Since the very high instantaneous energy is deposited on the heavy molten target in a very short period time, pressure waves are generated. They travel through the liquid and cause high stress in the container. Also, additional stress should be considered in the wall which is the result of direct heating of the target window. These dynamic processes were simulated with computational codes with the static response being analized first. The total resulting dynamic wall stress has been found to have exceeded themore » design stress for the selected container material. Adding a small amount of gas bubbles in the liquid could be a possible way to reduce the pressure waves.« less

  17. Curcumin targets fibroblast–tumor cell interactions in oral squamous cell carcinoma

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    Dudás, József, E-mail: jozsef.dudas@i-med.ac.at; Fullár, Alexandra, E-mail: fullarsz@gmail.com; 1st Department of Pathology and Experimental Cancer Research, Semmelweis University, Üllői út 26, 1085 Budapest

    Co-culture of periodontal ligament fibroblasts (PDLs) and SCC-25 oral squamous carcinoma cells (OSCC) results in conversion of PDLs into carcinoma-associated fibroblasts (CAFs) and induces epithelial-to mesenchymal transition (EMT) of OSCC tumor cells. We hypothesized that Curcumin targets this dynamic mutual interaction between CAFs and tumor cells. Normal and 2 μM Curcumin-treated co-culture were performed for 4 days, followed by analysis of tumor cell invasivity, mRNA/protein expression of EMT-markers and mediators, activity measure of matrix metalloproteinase 9 (MMP-9), and western blot analysis of signal transduction in tumor cells and fibroblasts. In Curcumin-treated co-culture, in tumor cells, the levels of nuclear factormore » κB (NFκBα) and early response kinase (ERK)—decreased, in fibroblasts, integrin αv protein synthesis decreased compared to corresponding cells in normal co-culture. The signal modulatory changes induced by Curcumin caused decreased release of EMT-mediators in CAFs and reversal of EMT in tumor cells, which was associated with decreased invasion. These data confirm the palliative potential of Curcumin in clinical application. - Graphical abstract: Co-culture of periodontal ligament fibroblasts (PDLs) and SCC-25 oral squamous carcinoma cells (OSCC) results in conversion of PDLs into carcinoma-associated fibroblasts (CAFs) and induces epithelial-to mesenchymal transition (EMT) of tumor cells. Curcumin targets this dynamic mutual interaction between CAFs and tumor cells by inhibiting the production of EMT mediators in CAFs and by modification of intracellular signaling in tumor cells. This causes less invasivity and reversal of EMT in tumor cells. Highlights: ► Curcumin targets tumor–fibroblast interaction in head and neck cancer. ► Curcumin suppresses mediators of epithelial–mesenchymal transition. ► Curcumin decreases the invasivity of tumor cells.« less

  18. Cells exposed to nanosecond electrical pulses exhibit biomarkers of mechanical stress

    NASA Astrophysics Data System (ADS)

    Roth, Caleb C.; Barnes, Ronald A.; Ibey, Bennett L.; Beier, Hope T.; Moen, Erick K.; Glickman, Randolph D.

    2015-03-01

    Exposure of cells to very short (<1 μs) electric pulses in the megavolt/meter range have been shown to cause disruption of the plasma membrane. This disruption is often characterized by the formation of numerous small pores (<2 nm in diameter) in the plasma membrane that last for several minutes, allowing the flow of ions into the cell. These small pores are called nanopores and the resulting damage to the plasma membrane is referred to as nanoporation. Nanosecond electrical pulse (nsEP) exposure can impart many different stressors on a cell, including electrical, electro-chemical, and mechanical stress. Thus, nsEP exposure is not a "clean" insult, making determination of the mechanism of nanoporation quite difficult. We hypothesize that nsEP exposure creates acoustic shock waves capable of causing nanoporation. Microarray analysis of primary adult human dermal fibroblasts (HDFa) exposed to nsEP, indicated several genes associated with mechanical stress were selectively upregulated 4 h post exposure. The idea that nanoporation is caused by external mechanical force from acoustic shock waves has, to our knowledge, not been investigated. This work will critically challenge the existing paradigm that nanoporation is caused solely by an electric-field driven event and could provide the basis for a plausible explanation for electroporation.

  19. Plasma Membrane Integrity and Survival of Melanoma Cells After Nanosecond Laser Pulses

    PubMed Central

    Pérez-Gutiérrez, Francisco G.; Camacho-López, Santiago; Evans, Rodger; Guillén, Gabriel; Goldschmidt, Benjamin S.; Viator, John A.

    2010-01-01

    Circulating tumor cells (CTCs) photoacoustic detection systems can aid clinical decision-making in the treatment of cancer. Interaction of melanin within melanoma cells with nanosecond laser pulses generates photoacoustic waves that make its detection possible. This study aims at: (1) determining melanoma cell survival after laser pulses of 6 ns at λ = 355 and 532 nm; (2) comparing the potential enhancement in the photoacoustic signal using λ = 355 nm in contrast with λ = 532 nm; (3) determining the critical laser fluence at which melanin begins to leak out from melanoma cells; and (4) developing a time-resolved imaging (TRI) system to study the intracellular interactions and their effect on the plasma membrane integrity. Monolayers of melanoma cells were grown on tissue culture-treated clusters and irradiated with up to 1.0 J/cm2. Surviving cells were stained with trypan blue and counted using a hemacytometer. The phosphate buffered saline absorbance was measured with a nanodrop spectrophotometer to detect melanin leakage from the melanoma cells post-laser irradiation. Photoacoustic signal magnitude was studied at both wavelengths using piezoelectric sensors. TRI with 6 ns resolution was used to image plasma membrane damage. Cell survival decreased proportionally with increasing laser fluence for both wavelengths, although the decrease is more pronounced for 355 nm radiation than for 532 nm. It was found that melanin leaks from cells equally for both wavelengths. No significant difference in photoacoustic signal was found between wavelengths. TRI showed clear damage to plasma membrane due to laser-induced bubble formation. PMID:20589533

  20. Quantifying pulsed electric field-induced membrane nanoporation in single cells.

    PubMed

    Moen, Erick K; Ibey, Bennett L; Beier, Hope T; Armani, Andrea M

    2016-11-01

    Plasma membrane disruption can trigger a host of cellular activities. One commonly observed type of disruption is pore formation. Molecular dynamic (MD) simulations of simplified lipid membrane structures predict that controllably disrupting the membrane via nano-scale poration may be possible with nanosecond pulsed electric fields (nsPEF). Until recently, researchers hoping to verify this hypothesis experimentally have been limited to measuring the relatively slow process of fluorescent markers diffusing across the membrane, which is indirect evidence of nanoporation that could be channel-mediated. Leveraging recent advances in nonlinear optical microscopy, we elucidate the role of pulse parameters in nsPEF-induced membrane permeabilization in live cells. Unlike previous techniques, it is able to directly observe loss of membrane order at the onset of the pulse. We also develop a complementary theoretical model that relates increasing membrane permeabilization to membrane pore density. Due to the significantly improved spatial and temporal resolution possible with our imaging method, we are able to directly compare our experimental and theoretical results. Their agreement provides substantial evidence that nanoporation does occur and that its development is dictated by the electric field distribution. Copyright © 2016 Elsevier B.V. All rights reserved.

  1. Cell-type-specific, Aptamer-functionalized Agents for Targeted Disease Therapy

    PubMed Central

    Zhou, Jiehua; Rossi, John J.

    2014-01-01

    One hundred years ago, Dr. Paul Ehrlich popularized the “magic bullet” concept for cancer therapy in which an ideal therapeutic agent would only kill the specific tumor cells it targeted. Since then, “targeted therapy” that specifically targets the molecular defects responsible for a patient's condition has become a long-standing goal for treating human disease. However, safe and efficient drug delivery during the treatment of cancer and infectious disease remains a major challenge for clinical translation and the development of new therapies. The advent of SELEX technology has inspired many groundbreaking studies that successfully adapted cell-specific aptamers for targeted delivery of active drug substances in both in vitro and in vivo models. By covalently linking or physically functionalizing the cell-specific aptamers with therapeutic agents, such as siRNA, microRNA, chemotherapeutics or toxins, or delivery vehicles, such as organic or inorganic nanocarriers, the targeted cells and tissues can be specifically recognized and the therapeutic compounds internalized, thereby improving the local concentration of the drug and its therapeutic efficacy. Currently, many cell-type-specific aptamers have been developed that can target distinct diseases or tissues in a cell-type-specific manner. In this review, we discuss recent advances in the use of cell-specific aptamers for targeted disease therapy, as well as conjugation strategies and challenges. PMID:24936916

  2. Trispecific antibodies for CD16A-directed NK cell engagement and dual-targeting of tumor cells.

    PubMed

    Gantke, Thorsten; Weichel, Michael; Herbrecht, Carmen; Reusch, Uwe; Ellwanger, Kristina; Fucek, Ivica; Eser, Markus; Müller, Thomas; Griep, Remko; Molkenthin, Vera; Zhukovsky, Eugene A; Treder, Martin

    2017-09-01

    Bispecific antibodies that redirect the lytic activity of cytotoxic immune effector cells, such as T- and NK cells, onto tumor cells have emerged as a highly attractive and clinically validated treatment modality for hematological malignancies. Advancement of this therapeutic concept into solid tumor indications, however, is hampered by the scarcity of targetable antigens that are surface-expressed on tumor cells but demonstrate only limited expression on healthy tissues. To overcome this limitation, the concept of dual-targeting, i.e. the simultaneous targeting of two tumor-expressed surface antigens with limited co-expression on non-malignant cells, with multispecific antibodies has been proposed to increase tumor selectivity of antibody-induced effector cell cytotoxicity. Here, a novel CD16A (FcγRIIIa)-directed trispecific, tetravalent antibody format, termed aTriFlex, is described, that is capable of redirecting NK cell cytotoxicity to two surface-expressed antigens. Using a BCMA/CD200-based in vitro model system, the potential use of aTriFlex antibodies for dual-targeting and selective induction of NK cell-mediated target cell lysis was investigated. Bivalent bispecific target cell binding was found to result in significant avidity gains and up to 17-fold increased in vitro potency. These data suggest trispecific aTriFlex antibodies may support dual-targeting strategies to redirect NK cell cytotoxicity with increased selectivity to enable targeting of solid tumor antigens. © The Author 2017. Published by Oxford University Press. All rights reserved. For Permissions, please e-mail: journals.permissions@oup.com.

  3. HTLV-1 Tax-Specific CTL Epitope-Pulsed Dendritic Cell Therapy Reduces Proviral Load in Infected Rats with Immune Tolerance against Tax.

    PubMed

    Ando, Satomi; Hasegawa, Atsuhiko; Murakami, Yuji; Zeng, Na; Takatsuka, Natsuko; Maeda, Yasuhiro; Masuda, Takao; Suehiro, Youko; Kannagi, Mari

    2017-02-01

    Adult T cell leukemia/lymphoma (ATL), a CD4 + T cell malignancy with a poor prognosis, is caused by human T cell leukemia virus type 1 (HTLV-1) infection. High proviral load (PVL) is a risk factor for the progression to ATL. We previously reported that some asymptomatic carriers had severely reduced functions of CTLs against HTLV-1 Tax, the major target Ag. Furthermore, the CTL responses tended to be inversely correlated with PVL, suggesting that weak HTLV-1-specific CTL responses may be involved in the elevation of PVL. Our previous animal studies indicated that oral HTLV-1 infection, the major route of infection, caused persistent infection with higher PVL in rats compared with other routes. In this study, we found that Tax-specific CD8 + T cells were present, but not functional, in orally infected rats as observed in some human asymptomatic carriers. Even in the infected rats with immune unresponsiveness against Tax, Tax-specific CTL epitope-pulsed dendritic cell (DC) therapy reduced the PVL and induced Tax-specific CD8 + T cells capable of proliferating and producing IFN-γ. Furthermore, we found that monocyte-derived DCs from most infected individuals still had the capacity to stimulate CMV-specific autologous CTLs in vitro, indicating that DC therapy may be applicable to most infected individuals. These data suggest that peptide-pulsed DC immunotherapy will be useful to induce functional HTLV-1-specific CTLs and decrease PVL in infected individuals with high PVL and impaired HTLV-1-specific CTL responses, thereby reducing the risk of the development of ATL. Copyright © 2017 by The American Association of Immunologists, Inc.

  4. Dendritic cell targeted vaccines: Recent progresses and challenges

    PubMed Central

    Chen, Pengfei; Liu, Xinsheng; Sun, Yuefeng; Zhou, Peng; Wang, Yonglu; Zhang, Yongguang

    2016-01-01

    ABSTRACT Dendritic cells (DCs) are known to be a set of morphology, structure and function of heterogeneous professional antigen presenting cells (APCs), as well as the strongest functional antigen presenting cells, which can absorb, process and present antigens. As the key regulators of innate and adaptive immune responses, DCs are at the center of the immune system and capable of interacting with both B cells and T cells, thereby manipulating the humoral and cellular immune responses. DCs provide an essential link between the innate and adaptive immunity, and the strong immune activation function of DCs and their properties of natural adjuvants, make them a valuable target for antigen delivery. Targeting antigens to DC-specific endocytic receptors in combination with the relevant antibodies or ligands along with immunostimulatory adjuvants has been recently recognized as a promising strategy for designing an effective vaccine that elicits a strong and durable T cell response against intracellular pathogens and cancer. This opinion article provides a brief summary of the rationales, superiorities and challenges of existing DC-targeting approaches. PMID:26513200

  5. Selective tumor cell targeting by the disaccharide moiety of bleomycin.

    PubMed

    Yu, Zhiqiang; Schmaltz, Ryan M; Bozeman, Trevor C; Paul, Rakesh; Rishel, Michael J; Tsosie, Krystal S; Hecht, Sidney M

    2013-02-27

    In a recent study, the well-documented tumor targeting properties of the antitumor agent bleomycin (BLM) were studied in cell culture using microbubbles that had been derivatized with multiple copies of BLM. It was shown that BLM selectively targeted MCF-7 human breast carcinoma cells but not the "normal" breast cell line MCF-10A. Furthermore, it was found that the BLM analogue deglycobleomycin, which lacks the disaccharide moiety of BLM, did not target either cell line, indicating that the BLM disaccharide moiety is necessary for tumor selectivity. Not resolved in the earlier study were the issues of whether the BLM disaccharide moiety alone is sufficient for tumor cell targeting and the possible cellular uptake of the disaccharide. In the present study, we conjugated BLM, deglycoBLM, and BLM disaccharide to the cyanine dye Cy5**. It was found that the BLM and BLM disaccharide conjugates, but not the deglycoBLM conjugate, bound selectively to MCF-7 cells and were internalized. The same was also true for the prostate cancer cell line DU-145 (but not for normal PZ-HPV-7 prostate cells) and for the pancreatic cancer cell line BxPC-3 (but not for normal SVR A221a pancreas cells). The targeting efficiency of the disaccharide was only slightly less than that of BLM in MCF-7 and DU-145 cells and comparable to that of BLM in BxPC-3 cells. These results establish that the BLM disaccharide is both necessary and sufficient for tumor cell targeting, a finding with obvious implications for the design of novel tumor imaging and therapeutic agents.

  6. An accelerated framework for the classification of biological targets from solid-state micropore data.

    PubMed

    Hanif, Madiha; Hafeez, Abdul; Suleman, Yusuf; Mustafa Rafique, M; Butt, Ali R; Iqbal, Samir M

    2016-10-01

    Micro- and nanoscale systems have provided means to detect biological targets, such as DNA, proteins, and human cells, at ultrahigh sensitivity. However, these devices suffer from noise in the raw data, which continues to be significant as newer and devices that are more sensitive produce an increasing amount of data that needs to be analyzed. An important dimension that is often discounted in these systems is the ability to quickly process the measured data for an instant feedback. Realizing and developing algorithms for the accurate detection and classification of biological targets in realtime is vital. Toward this end, we describe a supervised machine-learning approach that records single cell events (pulses), computes useful pulse features, and classifies the future patterns into their respective types, such as cancerous/non-cancerous cells based on the training data. The approach detects cells with an accuracy of 70% from the raw data followed by an accurate classification when larger training sets are employed. The parallel implementation of the algorithm on graphics processing unit (GPU) demonstrates a speedup of three to four folds as compared to a serial implementation on an Intel Core i7 processor. This incredibly efficient GPU system is an effort to streamline the analysis of pulse data in an academic setting. This paper presents for the first time ever, a non-commercial technique using a GPU system for realtime analysis, paired with biological cluster targeting analysis. Copyright © 2016 Elsevier Ireland Ltd. All rights reserved.

  7. Selective gene transfection of individual cells in vitro with plasmonic nanobubbles.

    PubMed

    Lukianova-Hleb, Ekaterina Y; Samaniego, Adam P; Wen, Jianguo; Metelitsa, Leonid S; Chang, Chung-Che; Lapotko, Dmitri O

    2011-06-10

    Gene delivery and transfection of eukaryotic cells are widely used for research and for developing gene cell therapy. However, the existing methods lack selectivity, efficacy and safety when heterogeneous cell systems must be treated. We report a new method that employs plasmonic nanobubbles (PNBs) for delivery and transfection. A PNB is a novel, tunable cellular agent with a dual mechanical and optical action due to the formation of the vapor nanobubble around a transiently heated gold nanoparticle upon its exposure to a laser pulse. PNBs enabled the mechanical injection of the extracellular cDNA plasmid into the cytoplasm of individual target living cells, cultured leukemia cells and human CD34+ CD117+ stem cells and expression of a green fluorescent protein (GFP) in those cells. PNB generation and lifetime correlated with the expression of green fluorescent protein in PNB-treated cells. Optical scattering by PNBs additionally provided the detection of the target cells and the guidance of cDNA injection at single cell level. In both cell models PNBs demonstrated a gene transfection effect in a single pulse treatment with high selectivity, efficacy and safety. Thus, PNBs provided targeted gene delivery at the single cell level in a single pulse procedure that can be used for safe and effective gene therapy. Copyright © 2011 Elsevier B.V. All rights reserved.

  8. An innovative pre-targeting strategy for tumor cell specific imaging and therapy.

    PubMed

    Qin, Si-Yong; Peng, Meng-Yun; Rong, Lei; Jia, Hui-Zhen; Chen, Si; Cheng, Si-Xue; Feng, Jun; Zhang, Xian-Zheng

    2015-09-21

    A programmed pre-targeting system for tumor cell imaging and targeting therapy was established based on the "biotin-avidin" interaction. In this programmed functional system, transferrin-biotin can be actively captured by tumor cells with the overexpression of transferrin receptors, thus achieving the pre-targeting modality. Depending upon avidin-biotin recognition, the attachment of multivalent FITC-avidin to biotinylated tumor cells not only offered the rapid fluorescence labelling, but also endowed the pre-targeted cells with targeting sites for the specifically designed biotinylated peptide nano-drug. Owing to the successful pre-targeting, tumorous HepG2 and HeLa cells were effectively distinguished from the normal 3T3 cells via fluorescence imaging. In addition, the self-assembled peptide nano-drug resulted in enhanced cell apoptosis in the observed HepG2 cells. The tumor cell specific pre-targeting strategy is applicable for a variety of different imaging and therapeutic agents for tumor treatments.

  9. Pulsed laser deposition of chalcogenide sulfides from multi- and single-component targets: the non-stoichiometric material transfer

    NASA Astrophysics Data System (ADS)

    Schou, Jørgen; Gansukh, Mungunshagai; Ettlinger, Rebecca B.; Cazzaniga, Andrea; Grossberg, Maarja; Kauk-Kuusik, Marit; Canulescu, Stela

    2018-01-01

    The mass transfer from target to films is incongruent for chalcogenide sulfides in contrast to the expectations of pulsed laser deposition (PLD) as a stoichiometric film growth process. Films produced from a CZTS (Cu2ZnSnS4) multi-component target have no Cu below a fluence threshold of 0.2 J/cm2, and the Cu content is also very low at low fluence from a single-component target. Above this threshold, the Cu content in the films increases almost linearly up to a value above the stoichiometric value, while the ratio of the concentration of the other metals Zn to Sn (Zn/Sn) remains constant. Films of a similar material CTS (Cu2SnS3) have been produced by PLD from a CTS target and exhibits a similar trend in the same fluence region. The results are discussed on the basis of solid-state data and the existing data from the literature.

  10. Photothermal and photoacoustic processes of laser activated nano-thermolysis of cells

    NASA Astrophysics Data System (ADS)

    Lapotko, Dmitri; Lukianova, Ekaterina; Mitskevich, Pavel; Smolnikova, Victoria; Potapnev, Michail; Konopleva, Marina; Andreeff, Michael; Oraevsky, Alexander

    2007-02-01

    Laser Activated Nano-Thermolysis was recently proposed for selective damage of individual target (cancer) cells by pulsed laser induced microbubbles around superheated clusters of optically absorbing nanoparticles (NP). One of the clinical applications of this technology is the elimination of residual tumor cells from human blood and bone marrow. Clinical standards for the safety and efficacy of such procedure require the development and verification of highly selective and controllable mechanisms of cell killing. Our previous experiments showed that laser-induced microbubble is the main damaging factor in the case cell irradiation by short laser pulses above the threshold. Our current aim was to study the cell damage mechanisms and analyze selectivity and efficacy of cell damage as a function of NP parameters, NP-cell interaction conditions, and conditions of bubble generation around NP and NP clusters in cells. Generation of laser-induced bubbles around gold NP with diameters 10-250 nm was studied in Acute Myeloblast Leukemia (AML) cultures, normal stem and model K562 human cells. Short laser pulses (10 ns, 532 nm) were applied to those cells in vitro and the processes in cells were investigated with photothermal, fluorescent and atomic force microscopies and also with fluorescence flow cytometry. We have found that the best selectivity of cell damage is achieved by (1) forming large clusters of optically absorbing NP in target cells and (2) irradiating the cells with single laser pulses with the lowest fluence that can generate microbubble only around large clusters but not around single NP. Laser microbubbles with the lifetime from 20 ns to 2000 ns generated in individual cells caused damage and lysis of the cellular membrane and consequently cell death. Laser microbubbles did not damage normal cells around the damaged target (tumor) cell. Laser irradiation with equal fluence did not cause any damage of cells without accumulated NP clusters.

  11. Strategies to target non-T-cell HIV reservoirs.

    PubMed

    Sacha, Jonah B; Ndhlovu, Lishomwa C

    2016-07-01

    A central question for the HIV cure field is to determine new ways to target clinically relevant, latently and actively replicating HIV-infected cells beyond resting memory CD4 T cells, particularly in anatomical areas of low drug penetrability. HIV eradication strategies being positioned for targeting HIV for extinction in the CD4 T-cell compartment may also show promise in non-CD4 T-cells reservoirs. Furthermore, several exciting novel therapeutic approaches specifically focused on HIV clearance from non-CD4 T-cell populations are being developed. Although reservoir validity in these non-CD4 T cells continues to remain debated, this review will highlight recent advances and make an argument as to their clinical relevancy as we progress towards an HIV cure.

  12. Diagnosis of warm dense conditions in foil targets heated by intense femtosecond laser pulses using Kα imaging spectroscopy

    DOE PAGES

    Bae, L. J.; Zastrau, U.; Chung, H. -K.; ...

    2018-03-01

    Warm dense conditions in titanium foils irradiated with intense femtosecond laser pulses are diagnosed using an x-ray imaging spectroscopy technique. The line shapes of radially resolved titanium Kα spectra are measured with a toroidally bent GaAs crystal and an x-ray charge-coupled device. Measured spectra are compared with the K-shell emissions modeled using an atomic kinetics – spectroscopy simulation code. Kα line shapes are strongly affected by warm (5-40 eV) bulk electron temperatures and imply multiple temperature distributions in the targets. Finally, the spatial distribution of temperature is dependent on the target thickness, and a thin target shows an advantage tomore » generate uniform warm dense conditions in a large area.« less

  13. Diagnosis of warm dense conditions in foil targets heated by intense femtosecond laser pulses using Kα imaging spectroscopy

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    Bae, L. J.; Zastrau, U.; Chung, H. -K.

    Warm dense conditions in titanium foils irradiated with intense femtosecond laser pulses are diagnosed using an x-ray imaging spectroscopy technique. The line shapes of radially resolved titanium Kα spectra are measured with a toroidally bent GaAs crystal and an x-ray charge-coupled device. Measured spectra are compared with the K-shell emissions modeled using an atomic kinetics – spectroscopy simulation code. Kα line shapes are strongly affected by warm (5-40 eV) bulk electron temperatures and imply multiple temperature distributions in the targets. Finally, the spatial distribution of temperature is dependent on the target thickness, and a thin target shows an advantage tomore » generate uniform warm dense conditions in a large area.« less

  14. Optical reprogramming of human somatic cells using ultrashort Bessel-shaped near-infrared femtosecond laser pulses

    NASA Astrophysics Data System (ADS)

    Uchugonova, Aisada; Breunig, Hans Georg; Batista, Ana; König, Karsten

    2015-11-01

    We report a virus-free optical approach to human cell reprogramming into induced pluripotent stem cells with low-power nanoporation using ultrashort Bessel-shaped laser pulses. Picojoule near-infrared sub-20 fs laser pulses at a high 85 MHz repetition frequency are employed to generate transient nanopores in the membrane of dermal fibroblasts for the introduction of four transcription factors to induce the reprogramming process. In contrast to conventional approaches which utilize retro- or lentiviruses to deliver genes or transcription factors into the host genome, the laser method is virus-free; hence, the risk of virus-induced cancer generation limiting clinical application is avoided.

  15. Potential targets for lung squamous cell carcinoma

    Cancer.gov

    Researchers have identified potential therapeutic targets in lung squamous cell carcinoma, the second most common form of lung cancer. The Cancer Genome Atlas (TCGA) Research Network study comprehensively characterized the lung squamous cell carcinoma gen

  16. N-acetylgalactosamine-functionalized dendrimers as hepatic cancer cell-targeted carriers.

    PubMed

    Medina, Scott H; Tekumalla, Venkatesh; Chevliakov, Maxim V; Shewach, Donna S; Ensminger, William D; El-Sayed, Mohamed E H

    2011-06-01

    There is an urgent need for novel polymeric carriers that can selectively deliver a large dose of chemotherapeutic agents into hepatic cancer cells to achieve high therapeutic activity with minimal systemic side effects. PAMAM dendrimers are characterized by a unique branching architecture and a large number of chemical surface groups suitable for coupling of chemotherapeutic agents. In this article, we report the coupling of N-acetylgalactosamine (NAcGal) to generation 5 (G5) of poly(amidoamine) (PAMAM-NH₂) dendrimers via peptide and thiourea linkages to prepare NAcGal-targeted carriers used for targeted delivery of chemotherapeutic agents into hepatic cancer cells. We describe the uptake of NAcGal-targeted and non-targeted G5 dendrimers into hepatic cancer cells (HepG2) as a function of G5 concentration and incubation time. We examine the contribution of the asialoglycoprotein receptor (ASGPR) to the internalization of NAcGal-targeted dendrimers into hepatic cancer cells through a competitive inhibition assay. Our results show that uptake of NAcGal-targeted G5 dendrimers into hepatic cancer cells occurs via ASGPR-mediated endocytosis. Internalization of these targeted carriers increased with the increase in G5 concentration and incubation time following Michaelis-Menten kinetics characteristic of receptor-mediated endocytosis. These results collectively indicate that G5-NAcGal conjugates function as targeted carriers for selective delivery of chemotherapeutic agents into hepatic cancer cells. Copyright © 2011 Elsevier Ltd. All rights reserved.

  17. Electric pulses used in electrochemotherapy and electrogene therapy do not significantly change the expression profile of genes involved in the development of cancer in malignant melanoma cells

    PubMed Central

    2009-01-01

    Background Electroporation is a versatile method for in vitro or in vivo delivery of different molecules into cells. However, no study so far has analysed the effects of electric pulses used in electrochemotherapy (ECT pulses) or electric pulses used in electrogene therapy (EGT pulses) on malignant cells. We studied the effect of ECT and EGT pulses on human malignant melanoma cells in vitro in order to understand and predict the possible effect of electric pulses on gene expression and their possible effect on cell behaviour. Methods We used microarrays with 2698 different oligonucleotides to obtain the expression profile of genes involved in apoptosis and cancer development in a malignant melanoma cell line (SK-MEL28) exposed to ECT pulses and EGT pulses. Results Cells exposed to ECT pulses showed a 68.8% average survival rate, while cells exposed to EGT pulses showed a 31.4% average survival rate. Only seven common genes were found differentially expressed in cells 16 h after exposure to ECT and EGT pulses. We found that ECT and EGT pulses induce an HSP70 stress response mechanism, repress histone protein H4, a major protein involved in chromatin assembly, and down-regulate components involved in protein synthesis. Conclusion Our results show that electroporation does not significantly change the expression profile of major tumour suppressor genes or oncogenes of the cell cycle. Moreover, electroporation also does not changes the expression of genes involved in the stability of DNA, supporting current evidence that electroporation is a safe method that does not promote tumorigenesis. However, in spite of being considered an isothermal method, it does to some extent induce stress, which resulted in the expression of the environmental stress response mechanism, HSP70. PMID:19709437

  18. Electric pulses used in electrochemotherapy and electrogene therapy do not significantly change the expression profile of genes involved in the development of cancer in malignant melanoma cells.

    PubMed

    Mlakar, Vid; Todorovic, Vesna; Cemazar, Maja; Glavac, Damjan; Sersa, Gregor

    2009-08-26

    Electroporation is a versatile method for in vitro or in vivo delivery of different molecules into cells. However, no study so far has analysed the effects of electric pulses used in electrochemotherapy (ECT pulses) or electric pulses used in electrogene therapy (EGT pulses) on malignant cells. We studied the effect of ECT and EGT pulses on human malignant melanoma cells in vitro in order to understand and predict the possible effect of electric pulses on gene expression and their possible effect on cell behaviour. We used microarrays with 2698 different oligonucleotides to obtain the expression profile of genes involved in apoptosis and cancer development in a malignant melanoma cell line (SK-MEL28) exposed to ECT pulses and EGT pulses. Cells exposed to ECT pulses showed a 68.8% average survival rate, while cells exposed to EGT pulses showed a 31.4% average survival rate. Only seven common genes were found differentially expressed in cells 16 h after exposure to ECT and EGT pulses. We found that ECT and EGT pulses induce an HSP70 stress response mechanism, repress histone protein H4, a major protein involved in chromatin assembly, and down-regulate components involved in protein synthesis. Our results show that electroporation does not significantly change the expression profile of major tumour suppressor genes or oncogenes of the cell cycle. Moreover, electroporation also does not changes the expression of genes involved in the stability of DNA, supporting current evidence that electroporation is a safe method that does not promote tumorigenesis. However, in spite of being considered an isothermal method, it does to some extent induce stress, which resulted in the expression of the environmental stress response mechanism, HSP70.

  19. Cell cycle proteins as promising targets in cancer therapy.

    PubMed

    Otto, Tobias; Sicinski, Piotr

    2017-01-27

    Cancer is characterized by uncontrolled tumour cell proliferation resulting from aberrant activity of various cell cycle proteins. Therefore, cell cycle regulators are considered attractive targets in cancer therapy. Intriguingly, animal models demonstrate that some of these proteins are not essential for proliferation of non-transformed cells and development of most tissues. By contrast, many cancers are uniquely dependent on these proteins and hence are selectively sensitive to their inhibition. After decades of research on the physiological functions of cell cycle proteins and their relevance for cancer, this knowledge recently translated into the first approved cancer therapeutic targeting of a direct regulator of the cell cycle. In this Review, we focus on proteins that directly regulate cell cycle progression (such as cyclin-dependent kinases (CDKs)), as well as checkpoint kinases, Aurora kinases and Polo-like kinases (PLKs). We discuss the role of cell cycle proteins in cancer, the rationale for targeting them in cancer treatment and results of clinical trials, as well as the future therapeutic potential of various cell cycle inhibitors.

  20. Manipulation of cell membrane using carbon nanotube scaffold as a photoresponsive stimuli generator

    PubMed Central

    Sada, Takao; Fujigaya, Tsuyohiko; Nakashima, Naotoshi

    2014-01-01

    We describe, for the first time, the perforation of the cell membrane in the targeted single cell based on the nanosecond pulsed near-infrared (NIR) laser irradiation of a thin film of carbon nanotubes that act as an effective photon absorber as well as stimuli generator. When the power of NIR laser is over 17.5 μJ/pulse, the cell membrane after irradiation is irreversibly disrupted and results in cell death. In sharp contrast, the perforation of the cell membrane occurs at suitable laser power (∼15 μJ/pulse) without involving cell termination. PMID:27877703

  1. FAST TRACK COMMUNICATION: Selective inactivation of micro-organisms with near-infrared femtosecond laser pulses

    NASA Astrophysics Data System (ADS)

    Tsen, K. T.; Tsen, Shaw-Wei D.; Sankey, Otto F.; Kiang, Juliann G.

    2007-11-01

    We demonstrate an unconventional and revolutionary method for selective inactivation of micro-organisms by using near-infrared femtosecond laser pulses. We show that if the wavelength and pulse width of the excitation femtosecond laser are appropriately selected, there exists a window in power density that enables us to achieve selective inactivation of target viruses and bacteria without causing cytotoxicity in mammalian cells. This strategy targets the mechanical (vibrational) properties of micro-organisms, and thus its antimicrobial efficacy is likely unaffected by genetic mutation in the micro-organisms. Such a method may be effective against a wide variety of drug resistant micro-organisms and has broad implications in disinfection as well as in the development of novel treatments for viral and bacterial pathogens.

  2. Folate-conjugated immunoglobulin targets melanoma tumor cells for NK cell effector functions

    PubMed Central

    Skinner, Cassandra C.; McMichael, Elizabeth L.; Jaime-Ramirez, Alena C.; Abrams, Zachary B.; Lee, Robert J.; Carson, William E.

    2016-01-01

    The folate receptor (FR) is over-expressed on the vascular side of cancerous cells including those of the breast, ovaries, testes, and cervix. We hypothesized that a folate-conjugated immunoglobulin (F-IgG) would bind to the FR that is over-expressed on melanoma tumor cells to target these cells for lysis by natural killer (NK) cells. Folate receptor expression was confirmed in the Mel-39 (human melanoma) cell line by flow cytometry and immunoblot analysis, using KB (human oral epithelial) and F01 (human melanoma) as a positive and negative control, respectively. FR-positive and negative cell lines were treated with F-IgG or control immunoglobulin G (C-IgG) in the presence or absence of cytokines in order to determine NK cell ability to lyse FR-positive cell lines. NK cell activation was significantly upregulated and lysis of Mel 39 tumor cells enhanced following treatment with F-IgG, as compared to C-IgG at all effector:target (E:T) ratios (p<0.01). This trend was further enhanced by NK cell stimulation with the activating cytokine interleukin-12 (IL-12). NK cell production of cytokines such as interferon-gamma (IFN-γ), macrophage inflammatory protein 1 alpha (MIP-1α), and regulated on activation normal T-cell expressed and secreted (RANTES) were also significantly increased in response to co-stimulation with IL-12 stimulation and F-IgG-coated Mel 39 target cells, as compared to controls (p<0.01). In contrast, F-IgG did not bind to the FR-negative cell line F01 and had no significant effect on NK cell lysis or cytokine production. This research indicates the potential use of F-IgG for its ability to induce an immune response from NK cells against FR-positive melanoma tumor cells which can be further enhanced by the addition of cytokines. PMID:27035691

  3. Pulse stretcher

    DOEpatents

    Horton, J.A.

    1994-05-03

    Apparatus for increasing the length of a laser pulse to reduce its peak power without substantial loss in the average power of the pulse is disclosed. The apparatus uses a White cell having a plurality of optical delay paths of successively increasing number of passes between the field mirror and the objective mirrors. A pulse from a laser travels through a multi-leg reflective path between a beam splitter and a totally reflective mirror to the laser output. The laser pulse is also simultaneously injected through the beam splitter to the input mirrors of the optical delay paths. The pulses from the output mirrors of the optical delay paths go simultaneously to the laser output and to the input mirrors of the longer optical delay paths. The beam splitter is 50% reflective and 50% transmissive to provide equal attenuation of all of the pulses at the laser output. 6 figures.

  4. Measurement of plasma momentum exerted on target by a small helicon plasma thruster and comparison with direct thrust measurement.

    PubMed

    Takahashi, Kazunori; Komuro, Atsushi; Ando, Akira

    2015-02-01

    Momentum, i.e., force, exerted from a small helicon plasma thruster to a target plate is measured simultaneously with a direct thrust measurement using a thrust balance. The calibration coefficient relating a target displacement to a steady-state force is obtained by supplying a dc to a calibration coil mounted on the target, where a force acting to a small permanent magnet located near the coil is directly measured by using a load cell. As the force exerted by the plasma flow to the target plate is in good agreement with the directly measured thrust, the validity of the target technique is demonstrated under the present operating conditions, where the thruster is operated in steady-state. Furthermore, a calibration coefficient relating a swing amplitude of the target to an impulse bit is also obtained by pulsing the calibration coil current. The force exerted by the pulsed plasma, which is estimated from the measured impulse bit and the pulse width, is also in good agreement with that obtained for the steady-state operation; hence, the thrust assessment of the helicon plasma thruster by the target is validated for both the steady-state and pulsed operations.

  5. Curcumin: a promising agent targeting cancer stem cells.

    PubMed

    Zang, Shufei; Liu, Tao; Shi, Junping; Qiao, Liang

    2014-01-01

    Cancer stem cells are a subset of cells that are responsible for cancer initiation and relapse. They are generally resistant to the current anticancer agents. Successful anticancer therapy must consist of approaches that can target not only the differentiated cancer cells, but also cancer stem cells. Emerging evidence suggested that the dietary agent curcumin exerted its anti-cancer activities via targeting cancer stem cells of various origins such as those of colorectal cancer, pancreatic cancer, breast cancer, brain cancer, and head and neck cancer. In order to enhance the therapeutic potential of curcumin, this agent has been modified or used in combination with other agents in the experimental therapy for many cancers. In this mini-review, we discussed the effect of curcumin and its derivatives in eliminating cancer stem cells and the possible underlying mechanisms.

  6. The mechanism of T-cell mediated cytotoxicity. VI. T-cell projections and their role in target cell killing.

    PubMed Central

    Sanderson, C J; Glauert, A M

    1979-01-01

    Electron micrographs of material fixed during the first 10 min of a T-cell cytotoxic system showed T-cell projections and T-cell burrowing into target cells. These observations were made possible by using a system with a very high rate of killing. The projections vary in shape and size, and can push deeply into the target cell, distorting organelles in their path, including the nucleus. The projections contain fine fibrillar material, to the exclusion of organelles. They push the target cell membrane in front of them to form pockets approximating to the shape of the projection. Areas of close contact occur between the projections and the target cell membrane, particularly at the leading edges. The likelihood that these projections develop as a result of contact with specific antigen, and are involved in the cytotoxic mechanism is discussed. Images Figure 1 Figure 2 Figure 3 Figure 4 Figure 5 Figure 6 Figure 7 Figure 8 Figure 9 Figure 10 Figure 11 Figure 12 Figure 13 Figure 14 Figure 15 Figure 16 PMID:311336

  7. Targeting survival pathways in chronic myeloid leukaemia stem cells

    PubMed Central

    Sinclair, A; Latif, A L; Holyoake, T L

    2013-01-01

    Chronic myeloid leukaemia (CML) is a clonal myeloproliferative disorder characterized by the presence of a fusion oncogene BCR-ABL, which encodes a protein with constitutive TK activity. The implementation of tyrosine kinase inhibitors (TKIs) marked a major advance in CML therapy; however, there are problems with current treatment. For example, relapse occurs when these drugs are discontinued in the majority of patients who have achieved a complete molecular response on TKI and these agents are less effective in patients with mutations in the BCR-ABL kinase domain. Importantly, TKI can effectively target proliferating mature cells, but do not eradicate quiescent leukaemic stem cells (LSCs), therefore allowing disease persistence despite treatment. It is essential that alternative strategies are used to target the LSC population. BCR-ABL activation is responsible for the modulation of different signalling pathways, which allows the LSC fraction to evade cell death. Several pathways have been shown to be modulated by BCR-ABL, including PI3K/AKT/mTOR, JAK-STAT and autophagy signalling pathways. Targeting components of these survival pathways, alone or in combination with TKI, therefore represents an attractive potential therapeutic approach for targeting the LSC. However, many pathways are also active in normal stem cells. Therefore, potential targets must be validated to effectively eradicate CML stem cells while sparing normal counterparts. This review summarizes the main pathways modulated in CML stem cells, the recent developments and the use of novel drugs to target components in these pathways which may be used to target the LSC population. Linked Articles This article is part of a themed section on Emerging Therapeutic Aspects in Oncology. To view the other articles in this section visit http://dx.doi.org/10.1111/bph.2013.169.issue-8 PMID:23517124

  8. Effect of dietary pulse intake on established therapeutic lipid targets for cardiovascular risk reduction: a systematic review and meta-analysis of randomized controlled trials

    PubMed Central

    Ha, Vanessa; Sievenpiper, John L.; de Souza, Russell J.; Jayalath, Viranda H.; Mirrahimi, Arash; Agarwal, Arnav; Chiavaroli, Laura; Mejia, Sonia Blanco; Sacks, Frank M.; Di Buono, Marco; Bernstein, Adam M.; Leiter, Lawrence A.; Kris-Etherton, Penny M.; Vuksan, Vladimir; Bazinet, Richard P.; Josse, Robert G.; Beyene, Joseph; Kendall, Cyril W.C.; Jenkins, David J.A.

    2014-01-01

    Background: Evidence from controlled trials encourages the intake of dietary pulses (beans, chickpeas, lentils and peas) as a method of improving dyslipidemia, but heart health guidelines have stopped short of ascribing specific benefits to this type of intervention or have graded the beneficial evidence as low. We conducted a systematic review and meta-analysis of randomized controlled trials (RCTs) to assess the effect of dietary pulse intake on established therapeutic lipid targets for cardiovascular risk reduction. Methods: We searched electronic databases and bibliographies of selected trials for relevant articles published through Feb. 5, 2014. We included RCTs of at least 3 weeks’ duration that compared a diet emphasizing dietary pulse intake with an isocaloric diet that did not include dietary pulses. The lipid targets investigated were low-density lipoprotein (LDL) cholesterol, apolipoprotein B and non–high-density lipoprotein (non-HDL) cholesterol. We pooled data using a random-effects model. Results: We identified 26 RCTs (n = 1037) that satisfied the inclusion criteria. Diets emphasizing dietary pulse intake at a median dose of 130 g/d (about 1 serving daily) significantly lowered LDL cholesterol levels compared with the control diets (mean difference −0.17 mmol/L, 95% confidence interval −0.25 to −0.09 mmol/L). Treatment effects on apolipoprotein B and non-HDL cholesterol were not observed. Interpretation: Our findings suggest that dietary pulse intake significantly reduces LDL cholesterol levels. Trials of longer duration and higher quality are needed to verify these results. Trial registration: ClinicalTrials.gov, no. NCT01594567. PMID:24710915

  9. Comparison study of a long-pulse pulsed dye laser and a long-pulse pulsed alexandrite laser in the treatment of port wine stains.

    PubMed

    Li, Li; Kono, Taro; Groff, William Frederick; Chan, Henry H; Kitazawa, Yoshihiko; Nozaki, Motohiro

    2008-03-01

    Port wine stains (PWSs) are commonly treated by the pulsed dye laser. Recently, a long-pulse pulsed alexandrite laser was used to treat bulky vascular malformations. In the present study, we compare the efficacy and complications of the long-pulse pulsed dye laser (LPPDL) and the long-pulse pulsed alexandrite laser (LPPAL) in the treatment of PWSs. Eleven patients with Fitzpatrick skin types III-IV were enrolled in this study. One section of each patient's PWS was treated with LPPDL and another section was treated with LPPAL. The patients' PWSs were evaluated for efficacy of elimination of erythema and for treatment-related side effects. Both LPPDL and LPPAL treatment are effective in the treatment of PWSs. Hyperpigmentation was seen in two areas treated with LPPDL and in three areas treated with LPPAL. Hypopigmentation was seen in one area treated with LPPAL, but not in any of the areas treated with LPPDL. There was no scarring. LPPAL works best with hypertrophic, purple PWSs, while LPPDL yields better clinical improvements with the flat, pink PWSs. Targeting of deoxyhemoglobin, deeper penetration, and higher fluence may explain the effectiveness of LPPAL in purple, hypertrophic PWSs. However, there is a risk of dyspigmentation when using the LPPAL.

  10. Breast cancer stem cells, EMT and therapeutic targets

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    Kotiyal, Srishti; Bhattacharya, Susinjan, E-mail: s.bhattacharya@jiit.ac.in

    Highlights: • Therapeutic targeting or inhibition of the key molecules of signaling pathways can control growth of breast cancer stem cells (BCSCs). • Development of BCSCs also involves miRNA interactions. • Therapeutic achievement can be done by targeting identified targets in the BCSC pathways. - Abstract: A small heterogeneous population of breast cancer cells acts as seeds to induce new tumor growth. These seeds or breast cancer stem cells (BCSCs) exhibit great phenotypical plasticity which allows them to undergo “epithelial to mesenchymal transition” (EMT) at the site of primary tumor and a future reverse transition. Apart from metastasis they aremore » also responsible for maintaining the tumor and conferring it with drug and radiation resistance and a tendency for post-treatment relapse. Many of the signaling pathways involved in induction of EMT are involved in CSC generation and regulation. Here we are briefly reviewing the mechanism of TGF-β, Wnt, Notch, TNF-α, NF-κB, RTK signalling pathways which are involved in EMT as well as BCSCs maintenance. Therapeutic targeting or inhibition of the key/accessory players of these pathways could control growth of BCSCs and hence malignant cancer. Additionally several miRNAs are dysregulated in cancer stem cells indicating their roles as oncogenes or tumor suppressors. This review also lists the miRNA interactions identified in BCSCs and discusses on some newly identified targets in the BCSC regulatory pathways like SHIP2, nicastrin, Pin 1, IGF-1R, pro-inflammatory cytokines and syndecan which can be targeted for therapeutic achievements.« less

  11. Splitter target for controlling magnetic reconnection in relativistic laser plasma interactions

    NASA Astrophysics Data System (ADS)

    Gu, Y. J.; Bulanov, S. S.; Korn, G.; Bulanov, S. V.

    2018-04-01

    The utilization of a conical target irradiated by a high power laser is proposed to study fast magnetic reconnection in relativistic plasma interactions. Such target, placed in front of the near critical density gas jet, splits the laser pulse, forming two parallel laser pulses in the 2D case and a donut shaped pulse in the 3D case. The magnetic annihilation and reconnection occur in the density downramp region of the subsequent gas jet. The magnetic field energy is converted into the particle kinetic energy. As a result, a backward accelerated electron beam is obtained as a signature of reconnection. The above mechanisms are demonstrated using particle-in-cell simulations in both 2D and 3D cases. Facilitating the synchronization of two laser beams, the proposed approach can be used in designing the corresponding experiments on studying fundamental problems of relativistic plasma physics.

  12. Plasma Membrane Permeabilization by Trains of Ultrashort Electric Pulses

    PubMed Central

    Ibey, Bennett L.; Mixon, Dustin G.; Payne, Jason A.; Bowman, Angela; Sickendick, Karl; Wilmink, Gerald J.; Roach, W. Patrick; Pakhomov, Andrei G.

    2010-01-01

    Ultrashort electric pulses (USEP) cause long-lasting increase of cell membrane electrical conductance, and that a single USEP increased cell membrane electrical conductance proportionally to the absorbed dose (AD) with a threshold of about 10 mJ/g. The present study extends quantification of the membrane permeabilization effect to multiple USEP and employed a more accurate protocol that identified USEP effect as the difference between post- and pre-exposure conductance values (Δg) in individual cells. We showed that Δg can be increased by either increasing the number of pulses at a constant E-field, or by increasing the E-field at a constant number of pulses. For 60-ns pulses, an E-field threshold of 6 kV/cm for a single pulse was lowered to less than 1.7 kV/cm by applying 100-pulse or longer trains. However, the reduction of the E-field threshold was only achieved at the expense of a higher AD compared to a single pulse exposure. Furthermore, the effect of multiple pulses was not fully determined by AD, suggesting that cells permeabilized by the first pulse(s) in the train become less vulnerable to subsequent pulses. This explanation was corroborated by a model that treated multiple-pulse exposures as a series of single-pulse exposures and assumed an exponential decline of cell susceptibility to USEP as Δg increased after each pulse during the course of the train. PMID:20171148

  13. The Mechanism of Gene Targeting in Human Somatic Cells

    PubMed Central

    Kan, Yinan; Ruis, Brian; Lin, Sherry; Hendrickson, Eric A.

    2014-01-01

    Gene targeting in human somatic cells is of importance because it can be used to either delineate the loss-of-function phenotype of a gene or correct a mutated gene back to wild-type. Both of these outcomes require a form of DNA double-strand break (DSB) repair known as homologous recombination (HR). The mechanism of HR leading to gene targeting, however, is not well understood in human cells. Here, we demonstrate that a two-end, ends-out HR intermediate is valid for human gene targeting. Furthermore, the resolution step of this intermediate occurs via the classic DSB repair model of HR while synthesis-dependent strand annealing and Holliday Junction dissolution are, at best, minor pathways. Moreover, and in contrast to other systems, the positions of Holliday Junction resolution are evenly distributed along the homology arms of the targeting vector. Most unexpectedly, we demonstrate that when a meganuclease is used to introduce a chromosomal DSB to augment gene targeting, the mechanism of gene targeting is inverted to an ends-in process. Finally, we demonstrate that the anti-recombination activity of mismatch repair is a significant impediment to gene targeting. These observations significantly advance our understanding of HR and gene targeting in human cells. PMID:24699519

  14. Pro-Tumoral Inflammatory Myeloid Cells as Emerging Therapeutic Targets.

    PubMed

    Szebeni, Gabor J; Vizler, Csaba; Nagy, Lajos I; Kitajka, Klara; Puskas, Laszlo G

    2016-11-23

    Since the observation of Virchow, it has long been known that the tumor microenvironment constitutes the soil for the infiltration of inflammatory cells and for the release of inflammatory mediators. Under certain circumstances, inflammation remains unresolved and promotes cancer development. Here, we review some of these indisputable experimental and clinical evidences of cancer related smouldering inflammation. The most common myeloid infiltrate in solid tumors is composed of myeloid-derived suppressor cells (MDSCs) and tumor-associated macrophages (TAMs). These cells promote tumor growth by several mechanisms, including their inherent immunosuppressive activity, promotion of neoangiogenesis, mediation of epithelial-mesenchymal transition and alteration of cellular metabolism. The pro-tumoral functions of TAMs and MDSCs are further enhanced by their cross-talk offering a myriad of potential anti-cancer therapeutic targets. We highlight these main pro-tumoral mechanisms of myeloid cells and give a general overview of their phenotypical and functional diversity, offering examples of possible therapeutic targets. Pharmacological targeting of inflammatory cells and molecular mediators may result in therapies improving patient condition and prognosis. Here, we review experimental and clinical findings on cancer-related inflammation with a major focus on creating an inventory of current small molecule-based therapeutic interventions targeting cancer-related inflammatory cells: TAMs and MDSCs.

  15. From cells to embryos: the application of femtosecond laser pulses for altering cellular material in complex biological systems

    NASA Astrophysics Data System (ADS)

    Kohli, V.; Elezzabi, A. Y.

    2008-02-01

    We report the application of high-intensity femtosecond laser pulses as a novel tool for manipulating biological specimens. When femtosecond laser pulses were focused to a near diffraction-limited focal spot, cellular material within the laser focal volume was surgically ablated. Several dissection cuts were made in the membrane of live mammalian cells, and membrane surgery was accomplished without inducing cell collapse or disassociation. By altering how the laser pulses were applied, focal adhesions joining live epithelial cells were surgically removed, resulting in single cell isolation. To further examine the versatility of this reported tool, cells were transiently permeabilized for introducing foreign material into the cytoplasm of live mammalian cells. Localizing focused femtosecond laser pulses on the biological membrane resulted in the formation of transient pores, which were harnessed as a pathway for the delivery of exogenous material. Individual mammalian cells were permeabilized in the presence of a hyperosmotic cryoprotective disaccharide. Material delivery was confirmed by measuring the volumetric response of cells permeabilized in 0.2, 0.3, 0.4 and 0.5 M cryoprotective sugar. The survival of permeabilized cells in increasing osmolarity of sugar was assessed using a membrane integrity assay. Further demonstrating the novelty of this reported tool, laser surgery of an aquatic embryo, the zebrafish (Danio rerio), was also performed. Utilizing the transient pores that were formed in the embryonic cells of the zebrafish embryo, an exogenous fluorescent probe FITC, Streptavidin-conjugated quantum dots or plasmid DNA (sCMV) encoding EGFP was introduced into the developing embryonic cells. To determine if the laser induced any short- or long-term effects on development, laser-manipulated embryos were reared to 2 and 7 days post-fertilization and compared to control embryos at the same developmental stages. Light microscopy and scanning electron microscopy

  16. Pulse shaping with transmission lines

    DOEpatents

    Wilcox, Russell B.

    1987-01-01

    A method and apparatus for forming shaped voltage pulses uses passive reflection from a transmission line with nonuniform impedance. The impedance of the reflecting line varies with length in accordance with the desired pulse shape. A high voltage input pulse is transmitted to the reflecting line. A reflected pulse is produced having the desired shape and is transmitted by pulse removal means to a load. Light activated photoconductive switches made of silicon can be utilized. The pulse shaper can be used to drive a Pockels cell to produce shaped optical pulses.

  17. Pulse shaping with transmission lines

    DOEpatents

    Wilcox, R.B.

    1985-08-15

    A method and apparatus for forming shaped voltage pulses uses passive reflection from a transmission line with nonuniform impedance. The impedance of the reflecting line varies with length in accordance with the desired pulse shape. A high voltage input pulse is transmitted to the reflecting line. A reflected pulse is produced having the desired shape and is transmitted by pulse removal means to a load. Light activated photoconductive switches made of silicon can be utilized. The pulse shaper can be used to drive a Pockels cell to produce shaped optical pulses.

  18. Prodrug strategy for cancer cell-specific targeting: A recent overview.

    PubMed

    Zhang, Xian; Li, Xiang; You, Qidong; Zhang, Xiaojin

    2017-10-20

    The increasing development of targeted cancer therapy provides extensive possibilities in clinical trials, and numerous strategies have been explored. The prodrug is one of the most promising strategies in targeted cancer therapy to improve the selectivity and efficacy of cytotoxic compounds. Compared with normal tissues, cancer cells are characterized by unique aberrant markers, thus inactive prodrugs targeting these markers are excellent therapeutics to release active drugs, killing cancer cells without damaging normal tissues. In this review, we explore an integrated view of potential prodrugs applied in targeted cancer therapy based on aberrant cancer specific markers and some examples are provided for inspiring new ideas of prodrug strategy for cancer cell-specific targeting. Copyright © 2017. Published by Elsevier Masson SAS.

  19. Design of a secondary ionization target for direct production of a C- beam from CO2 pulses for online AMS.

    PubMed

    Salazar, Gary; Ognibene, Ted

    2013-01-01

    We designed and optimized a novel device "target" that directs a CO 2 gas pulse onto a Ti surface where a Cs + beam generates C - from the CO 2 . This secondary ionization target enables an accelerator mass spectrometer to ionize pulses of CO 2 in the negative mode to measure 14 C/ 12 C isotopic ratios in real time. The design of the targets were based on computational flow dynamics, ionization mechanism and empirical optimization. As part of the ionization mechanism, the adsorption of CO 2 on the Ti surface was fitted with the Jovanovic-Freundlich isotherm model using empirical and simulation data. The inferred adsorption constants were in good agreement with other works. The empirical optimization showed that amount of injected carbon and the flow speed of the helium carrier gas improve the ionization efficiency and the amount of 12 C - produced until reaching a saturation point. Linear dynamic range between 150 and 1000 ng of C and optimum carrier gas flow speed of around 0.1 mL/min were shown. It was also shown that the ionization depends on the area of the Ti surface and Cs + beam cross-section. A range of ionization efficiency of 1-2.5% was obtained by optimizing the described parameters.

  20. Bystander Effect Induced by Electroporation is Possibly Mediated by Microvesicles and Dependent on Pulse Amplitude, Repetition Frequency and Cell Type.

    PubMed

    Prevc, Ajda; Bedina Zavec, Apolonija; Cemazar, Maja; Kloboves-Prevodnik, Veronika; Stimac, Monika; Todorovic, Vesna; Strojan, Primoz; Sersa, Gregor

    2016-10-01

    Bystander effect, a known phenomenon in radiation biology, where irradiated cells release signals which cause damage to nearby, unirradiated cells, has not been explored in electroporated cells yet. Therefore, our aim was to determine whether bystander effect is present in electroporated melanoma cells in vitro, by determining viability of non-electroporated cells exposed to medium from electroporated cells and by the release of microvesicles as potential indicators of the bystander effect. Here, we demonstrated that electroporation of cells induces bystander effect: Cells exposed to electric pulses mediated their damage to the non-electroporated cells, thus decreasing cell viability. We have shown that shedding microvesicles may be one of the ways used by the cells to mediate the death signals to the neighboring cells. The murine melanoma B16F1 cell line was found to be more electrosensitive and thus more prone to bystander effect than the canine melanoma CMeC-1 cell line. In B16F1 cell line, bystander effect was present above the level of electropermeabilization of the cells, with the threshold at 800 V/cm. Furthermore, with increasing electric field intensities and the number of pulses, the bystander effect also increased. In conclusion, electroporation can induce bystander effect which may be mediated by microvesicles, and depends on pulse amplitude, repetition frequency and cell type.

  1. In Situ Target Engagement Studies in Adherent Cells.

    PubMed

    Axelsson, Hanna; Almqvist, Helena; Otrocka, Magdalena; Vallin, Michaela; Lundqvist, Sara; Hansson, Pia; Karlsson, Ulla; Lundbäck, Thomas; Seashore-Ludlow, Brinton

    2018-04-20

    A prerequisite for successful drugs is effective binding of the desired target protein in the complex environment of a living system. Drug-target engagement has typically been difficult to monitor in physiologically relevant models, and with current methods, especially, while maintaining spatial information. One recent technique for quantifying drug-target engagement is the cellular thermal shift assay (CETSA), in which ligand-induced protein stabilization is measured after a heat challenge. Here, we describe a CETSA protocol in live A431 cells for p38α (MAPK14), where remaining soluble protein is detected in situ, using high-content imaging in 384-well, microtiter plates. We validate this assay concept using a number of known p38α inhibitors and further demonstrate the potential of this technology for chemical probe and drug discovery purposes by performing a small pilot screen for novel p38α binders. Importantly, this protocol creates a workflow that is amenable to adherent cells in their native state and yields spatially resolved target engagement information measurable at the single-cell level.

  2. Cytostatic response of NB69 cells to weak pulse-modulated 2.2 GHz radar-like signals.

    PubMed

    Trillo, María A; Cid, María Antonia; Martínez, Maria Antonia; Page, Juan E; Esteban, Jaime; Úbeda, Alejandro

    2011-07-01

    The present study investigates the response of two human cancer cell lines to a 24-h treatment with a 2.2-GHz, pulse-modulated (5 µs pulse duration, 100 Hz repetition rate) radar-like signal at an average SAR = 0.023 W/kg, using a newly designed setup for in vitro exposure to radiofrequency (RF) fields. A complete discretized model of the setup was created for numerical dosimetry using finite-difference time-domain (FDTD) software, SEMCAD X. The average dose of RF radiation absorbed by the cultures was calculated to be subthermal (ΔT < 0.1 °C). The RF exposure induced a consistent, statistically significant reduction in the cell number (13.5% below controls, P < 0.001) in the neuroblastoma NB69 line. This effect was accompanied with slight but statistically significant increases in the proportions of cells in phases G0/G1 and G2/M of the cell cycle (6% and 9%, respectively; P < 0.05 over controls). By contrast, the hepatocarcinoma cell line HepG2 did not respond to the same RF treatment. These results indicate that a pulse-modulated RF radiation with high instantaneous amplitude and low average power can induce cytostatic responses on specific, sensitive cancer cell lines. The effect would be mediated, at least in part, by alterations in the kinetics of the cell cycle. Copyright © 2011 Wiley-Liss, Inc.

  3. Femtosecond-laser assisted cell reprogramming

    NASA Astrophysics Data System (ADS)

    Breunig, Hans Georg; Uchugonova, Aisada; Batista, Ana; König, Karsten

    2017-02-01

    Femtosecond-laser pulses can assist to transfect cells by creating transient holes in the cell membrane, thus making them temporarily permeable for extraneous genetic material. This procedure offers the advantage of being completely "virus free" since no viruses are used for the delivery and integration of gene factors into the host genome and, thereby, avoiding serious side effects which so far prevent clinical application. Unfortunately, focusing of the laser radiation onto individual cell membranes is quite elaborate and time consuming. Regarding these obstacles, we briefly review two optical setups for fast, efficient and high throughput laser-assisted cell transfection based on femtosecond laser pulse excitation. The first setup aims at assisting the transfection of adherent cells. It comprises of a modified laser-scanning microscope with beamshaping optics as well as home-made software to automate the detection, targeting and laser-irradiation process. The second setup aims at laser-assisted transfection of non-adherent cells in suspension which move in a continuous flow through the laser focus region. The setup allows to address a large number of cells, however, with much lower transfection efficiency than the individual-cell targeting approach.

  4. Hybrid Pulsed Nd:YAG Laser

    NASA Astrophysics Data System (ADS)

    Miller, Sawyer; Trujillo, Skyler; Fort Lewis College Laser Group Team

    This work concerns the novel design of an inexpensive pulsed Nd:YAG laser, consisting of a hybrid Kerr Mode Lock (KLM) and Q-switch pulse. The two pulse generation systems work independently, non simultaneously of each other, thus generating the ability for the user to easily switch between ultra-short pulse widths or large energy density pulses. Traditionally, SF57 glass has been used as the Kerr medium. In this work, novel Kerr mode-locking mediums are being investigated including: tellurite compound glass (TeO2), carbon disulfide (CS2), and chalcogenide glass. These materials have a nonlinear index of refraction orders of magnitude,(n2), larger than SF57 glass. The Q-switched pulse will utilize a Pockels cell. As the two pulse generation systems cannot be operated simultaneously, the Pockels cell and Kerr medium are attached to kinematic mounts, allowing for quick interchange between systems. Pulse widths and repetition rates will vary between the two systems. A goal of 100 picosecond pulse widths are desired for the mode-locked system. A goal of 10 nanosecond pulse widths are desired for the Q-switch system, with a desired repetition rate of 50 Hz. As designed, the laser will be useful in imaging applications.

  5. Fragments of Target Cells are Internalized into Retroviral Envelope Protein-Expressing Cells during Cell-Cell Fusion by Endocytosis

    PubMed Central

    Izumida, Mai; Kamiyama, Haruka; Suematsu, Takashi; Honda, Eri; Koizumi, Yosuke; Yasui, Kiyoshi; Hayashi, Hideki; Ariyoshi, Koya; Kubo, Yoshinao

    2016-01-01

    Retroviruses enter into host cells by fusion between viral and host cell membranes. Retroviral envelope glycoprotein (Env) induces the membrane fusion, and also mediates cell-cell fusion. There are two types of cell-cell fusions induced by the Env protein. Fusion-from-within is induced by fusion between viral fusogenic Env protein-expressing cells and susceptible cells, and virions induce fusion-from-without by fusion between adjacent cells. Although entry of ecotropic murine leukemia virus (E-MLV) requires host cell endocytosis, the involvement of endocytosis in cell fusion is unclear. By fluorescent microscopic analysis of the fusion-from-within, we found that fragments of target cells are internalized into Env-expressing cells. Treatment of the Env-expressing cells with an endocytosis inhibitor more significantly inhibited the cell fusion than that of the target cells, indicating that endocytosis in Env-expressing cells is required for the cell fusion. The endocytosis inhibitor also attenuated the fusion-from-without. Electron microscopic analysis suggested that the membrane fusion resulting in fusion-from-within initiates in endocytic membrane dents. This study shows that two types of the viral cell fusion both require endocytosis, and provides the cascade of fusion-from-within. PMID:26834711

  6. Short-pulse, compressed ion beams at the Neutralized Drift Compression Experiment

    DOE PAGES

    Seidl, P. A.; Barnard, J. J.; Davidson, R. C.; ...

    2016-05-01

    We have commenced experiments with intense short pulses of ion beams on the Neutralized Drift Compression Experiment (NDCX-II) at Lawrence Berkeley National Laboratory, with 1-mm beam spot size within 2.5 ns full-width at half maximum. The ion kinetic energy is 1.2 MeV. To enable the short pulse duration and mm-scale focal spot radius, the beam is neutralized in a 1.5-meter-long drift compression section following the last accelerator cell. A short-focal-length solenoid focuses the beam in the presence of the volumetric plasma that is near the target. In the accelerator, the line-charge density increases due to the velocity ramp imparted onmore » the beam bunch. The scientific topics to be explored are warm dense matter, the dynamics of radiation damage in materials, and intense beam and beam-plasma physics including select topics of relevance to the development of heavy-ion drivers for inertial fusion energy. Below the transition to melting, the short beam pulses offer an opportunity to study the multi-scale dynamics of radiation-induced damage in materials with pump-probe experiments, and to stabilize novel metastable phases of materials when short-pulse heating is followed by rapid quenching. First experiments used a lithium ion source; a new plasma-based helium ion source shows much greater charge delivered to the target.« less

  7. Nanosecond pulsed electric fields induce poly(ADP-ribose) formation and non-apoptotic cell death in HeLa S3 cells

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    Morotomi-Yano, Keiko; Akiyama, Hidenori; Yano, Ken-ichi, E-mail: yanoken@kumamoto-u.ac.jp

    Highlights: •Nanosecond pulsed electric field (nsPEF) is a new and unique means for life sciences. •Apoptosis was induced by nsPEF exposure in Jurkat cells. •No signs of apoptosis were detected in HeLa S3 cells exposed to nsPEFs. •Formation of poly(ADP-ribose) was induced in nsPEF-exposed HeLa S3 cells. •Two distinct modes of cell death were activated by nsPEF in a cell-dependent manner. -- Abstract: Nanosecond pulsed electric fields (nsPEFs) have recently gained attention as effective cancer therapy owing to their potency for cell death induction. Previous studies have shown that apoptosis is a predominant mode of nsPEF-induced cell death in severalmore » cell lines, such as Jurkat cells. In this study, we analyzed molecular mechanisms for cell death induced by nsPEFs. When nsPEFs were applied to Jurkat cells, apoptosis was readily induced. Next, we used HeLa S3 cells and analyzed apoptotic events. Contrary to our expectation, nsPEF-exposed HeLa S3 cells exhibited no molecular signs of apoptosis execution. Instead, nsPEFs induced the formation of poly(ADP-ribose) (PAR), a hallmark of necrosis. PAR formation occurred concurrently with a decrease in cell viability, supporting implications of nsPEF-induced PAR formation for cell death. Necrotic PAR formation is known to be catalyzed by poly(ADP-ribose) polymerase-1 (PARP-1), and PARP-1 in apoptotic cells is inactivated by caspase-mediated proteolysis. Consistently, we observed intact and cleaved forms of PARP-1 in nsPEF-exposed and UV-irradiated cells, respectively. Taken together, nsPEFs induce two distinct modes of cell death in a cell type-specific manner, and HeLa S3 cells show PAR-associated non-apoptotic cell death in response to nsPEFs.« less

  8. The therapeutic potential of cell cycle targeting in multiple myeloma.

    PubMed

    Maes, Anke; Menu, Eline; Veirman, Kim De; Maes, Ken; Vand Erkerken, Karin; De Bruyne, Elke

    2017-10-27

    Proper cell cycle progression through the interphase and mitosis is regulated by coordinated activation of important cell cycle proteins (including cyclin-dependent kinases and mitotic kinases) and several checkpoint pathways. Aberrant activity of these cell cycle proteins and checkpoint pathways results in deregulation of cell cycle progression, which is one of the key hallmarks of cancer. Consequently, intensive research on targeting these cell cycle regulatory proteins identified several candidate small molecule inhibitors that are able to induce cell cycle arrest and even apoptosis in cancer cells. Importantly, several of these cell cycle regulatory proteins have also been proposed as therapeutic targets in the plasma cell malignancy multiple myeloma (MM). Despite the enormous progress in the treatment of MM the past 5 years, MM still remains most often incurable due to the development of drug resistance. Deregulated expression of the cyclins D is observed in virtually all myeloma patients, emphasizing the potential therapeutic interest of cyclin-dependent kinase inhibitors in MM. Furthermore, other targets have also been identified in MM, such as microtubules, kinesin motor proteins, aurora kinases, polo-like kinases and the anaphase promoting complex/cyclosome. This review will provide an overview of the cell cycle proteins and checkpoint pathways deregulated in MM and discuss the therapeutic potential of targeting proteins or protein complexes involved in cell cycle control in MM.

  9. Target Glint Suppression Technology.

    DTIC Science & Technology

    1980-09-01

    report is organized into two principal sections. Section 2 addresses the impact of target effects on the noncoherent detection problem associated with...zero pulse-to-pulse correlation. Results are presented for a scanning search radar which is assumed to noncoherently integrate N pulses. Generally...speaking, detection performance is shown to be a maximum when the pulse-to-pulse correlation is a minimum. As a result noncoherent search radars should

  10. Development of the dense plasma focus for short-pulse applications

    NASA Astrophysics Data System (ADS)

    Bennett, N.; Blasco, M.; Breeding, K.; Constantino, D.; DeYoung, A.; DiPuccio, V.; Friedman, J.; Gall, B.; Gardner, S.; Gatling, J.; Hagen, E. C.; Luttman, A.; Meehan, B. T.; Misch, M.; Molnar, S.; Morgan, G.; O'Brien, R.; Robbins, L.; Rundberg, R.; Sipe, N.; Welch, D. R.; Yuan, V.

    2017-01-01

    The dense plasma focus (DPF) has long been considered a compact source for pulsed neutrons and has traditionally been optimized for the total neutron yield. In this paper, we describe the efforts to optimize the DPF for short-pulse applications by introducing a reentrant cathode at the end of the coaxial plasma gun. The resulting neutron pulse widths are reduced by an average of 21 ±9 % from the traditional long-drift DPF design. Pulse widths and yields achieved from deuterium-tritium fusion at 2 MA are 61.8 ±30.7 ns FWHM and 1.84 ±0.49 ×1012 neutrons per shot. Simulations were conducted concurrently to elucidate the DPF operation and confirm the role of the reentrant cathode. A hybrid fluid-kinetic particle-in-cell modeling capability demonstrates correct sheath velocities, plasma instabilities, and fusion yield rates. Consistent with previous findings that the DPF is dominated by beam-target fusion from superthermal ions, we estimate that the thermonuclear contribution is at the 1% level.

  11. Cancer stem cell-targeted therapeutics and delivery strategies.

    PubMed

    Ahmad, Gulzar; Amiji, Mansoor M

    2017-08-01

    Cancer initiating or stem cells (CSCs) are a small population of cells in the tumor mass, which have been reported to be present in different types of cancers. CSCs usually reside within the tumor and are responsible for reoccurrence of cancer. The imprecise, inaccessible nature and increased efflux of conventional therapeutic drugs make these cells resistant to drugs. We discuss the specific markers for identification of these cells, role of CSCs in chemotherapy resistance and use of different therapeutic means to target them, including elucidation of specific cell markers, exploitation of different signaling pathways and use of nanotechnology. Area covered: This review covers cancer stem cell signaling which are used by these cells to maintain their quiescence, stemness and resistant phenotype, distinct cell surface markers, contribution of these cells in drug resistance, inevitability to cure cancer and use of nanotechnology to overcome this hurdle. Expert opinion: Cancer stem cells are the main culprit of our failure to cure cancer. In order to cure cancer along with other cells types in cancer, cancer stem cells need to be targeted in the tumor bed. Nanotechnology solutions can facilitate clinical translation of the therapeutics along with other emerging technologies to cure cancer.

  12. [EFFECT OF PULSE-PERIODIC CORONA DISCHARGE ON VIABILITY OF ESCHERICHIA COLI M17 CELLS IN BIOFILMS].

    PubMed

    Rybalchenko, O V; Stepanova, O M; Orlova, O G; Astafiev, A M; Kudryavtsev, A A; Kapustina, V V

    2015-01-01

    Detection of bactericidal effect of pulse-periodic corona discharge (PPCD) on cells and biofilms of Escherichia coli M17. A gas-discharge device was created based on PPCD in air with power supply parameters: amplitude values of voltage of 30 - 60 kV, pulse repetition rate of 250 - 400 kHz. Ultrastructure changes in cells and biofilms of E. coli M17, affected by PPCD, generated in air, were studied by typical methods of transmission electron microscopy. Disturbances of integrity of surface and abyssal structures of biofilms, as well as changes of morphological properties of E. coli M17 cells, characteristic for sub-lethal heat impact, were detected. Destructive changes of bacterial cells were developed by formation of focal disturbance of cytoplasmic membrane, extension of periplasmic space, formation of globular structures, characteristic for heat effect, and destruction of cytoplasm. Bactericidal effect of PPCD on E. coli M17 cells as part of biofilms was shown. Destructive morphological changes in cells and biofilms of E. coli M17 after the effect of PPCD were detected for the first time on electron-microscopic level.

  13. Mechanisms that enhance sustainability of p53 pulses.

    PubMed

    Kim, Jae Kyoung; Jackson, Trachette L

    2013-01-01

    The tumor suppressor p53 protein shows various dynamic responses depending on the types and extent of cellular stresses. In particular, in response to DNA damage induced by γ-irradiation, cells generate a series of p53 pulses. Recent research has shown the importance of sustaining repeated p53 pulses for recovery from DNA damage. However, far too little attention has been paid to understanding how cells can sustain p53 pulses given the complexities of genetic heterogeneity and intrinsic noise. Here, we explore potential molecular mechanisms that enhance the sustainability of p53 pulses by developing a new mathematical model of the p53 regulatory system. This model can reproduce many experimental results that describe the dynamics of p53 pulses. By simulating the model both deterministically and stochastically, we found three potential mechanisms that improve the sustainability of p53 pulses: 1) the recently identified positive feedback loop between p53 and Rorα allows cells to sustain p53 pulses with high amplitude over a wide range of conditions, 2) intrinsic noise can often prevent the dampening of p53 pulses even after mutations, and 3) coupling of p53 pulses in neighboring cells via cytochrome-c significantly reduces the chance of failure in sustaining p53 pulses in the presence of heterogeneity among cells. Finally, in light of these results, we propose testable experiments that can reveal important mechanisms underlying p53 dynamics.

  14. Surface Modification of ICF Target Capsules by Pulsed Laser Ablation

    DOE PAGES

    Carlson, Lane C.; Johnson, Michael A.; Bunn, Thomas L.

    2016-06-30

    Topographical modifications of spherical surfaces are imprinted on National Ignition Facility (NIF) target capsules by extending the capabilities of a recently developed full surface (4π) laser ablation and mapping apparatus. The laser ablation method combines the precision, energy density and long reach of a focused laser beam to pre-impose sinusoidal modulations on the outside surface of High Density Carbon (HDC) capsules and the inside surface of Glow Discharge Polymer (GDP) capsules. Sinusoidal modulations described in this paper have sub-micron to 10’s of microns vertical scale and wavelengths as small as 30 μm and as large as 200 μm. The modulatedmore » patterns are created by rastering a focused laser fired at discrete capsule surface locations for a specified number of pulses. The computer program developed to create these raster patterns uses inputs such as laser beam intensity profile, the material removal function, the starting surface figure and the desired surface figure. The patterns are optimized to minimize surface roughness. Lastly, in this paper, simulated surfaces are compared with actual ablated surfaces measured using confocal microscopy.« less

  15. Femtosecond laser pulse optimization for multiphoton cytometry and control of fluorescence

    NASA Astrophysics Data System (ADS)

    Tkaczyk, Eric Robert

    This body of work encompasses optimization of near infrared femtosecond laser pulses both for enhancement of flow cytometry as well as adaptive pulse shaping to control fluorescence. A two-photon system for in vivo flow cytometry is demonstrated, which allows noninvasive quantification of circulating cell populations in a single live mouse. We monitor fluorescently-labeled red blood cells for more than two weeks, and are also able to noninvasively measure circulation times of two distinct populations of breast cancer cells simultaneously in a single mouse. We build a custom laser excitation source in the form of an extended cavity mode-locked oscillator, which enables superior detection in whole blood or saline of cell lines expressing fluorescent proteins including the green fluorescent protein (GFP), tdTomato and mPlum. A mathematical model explains unique features of the signals. The ability to distinguish different fluorescent species is central to simultaneous measurement of multiple molecular targets in high throughput applications including the multiphoton flow cytometer. We demonstrate that two dyes which are not distinguishable to one-photon measurements can be differentiated and in fact quantified in mixture via phase-shaped two-photon excitation pulses found by a genetic algorithm. We also selectively enhance or suppress two-photon fluorescence of numerous common dyes with tailored pulse shapes. Using a multiplicative (rather than ratiometric) fitness parameter, we are able to control the fluorescence while maintaining a strong signal. With this method, we control the two-photon fluorescence of the blue fluorescent protein (BFP), which is of particular interest in investigations of protein-protein interactions, and has frustrated previous attempts of control. Implementing an acousto-optic interferometer, we use the same experimental setup to measure two-photon excitation cross-sections of dyes and prove that photon-photon interferences are the

  16. Pros and Cons of Antigen-Presenting Cell Targeted Tumor Vaccines.

    PubMed

    Goyvaerts, Cleo; Breckpot, Karine

    2015-01-01

    In therapeutic antitumor vaccination, dendritic cells play the leading role since they decide if, how, when, and where a potent antitumor immune response will take place. Since the disentanglement of the complexity and merit of different antigen-presenting cell subtypes, antitumor immunotherapeutic research started to investigate the potential benefit of targeting these subtypes in situ. This review will discuss which antigen-presenting cell subtypes are at play and how they have been targeted and finally question the true meaning of targeting antitumor-based vaccines.

  17. Progress toward a practical laser driven ion source using variable thickness liquid crystal targets

    NASA Astrophysics Data System (ADS)

    Poole, Patrick; Cochran, Ginevra; Zeil, Karl; Metzkes, Josephine; Obst, Lieselotte; Kluge, Thomas; Schlenvoigt, Hans-Peter; Prencipe, Irene; Cowan, Tom; Schramm, Uli; Schumacher, Douglass

    2016-10-01

    Ion acceleration from ultra-intense laser interaction has been long investigated in pursuit of requisite energies and spectral distributions for applications like proton cancer therapy. However, the details of ion acceleration mechanisms and their laser intensity scaling are not fully understood, especially the complete role of pulse contrast and target thickness. Additionally, target delivery and alignment at appropriate rates for study and subsequent treatment pose significant challenges. We present results from a campaign on the Draco laser using liquid crystal targets that have on-demand, in-situ thickness tunability over more than three orders of magnitude, enabling rapid data collection due to <1 minute, automatically aligned target formation. Diagnostics include spectral and spatial measurement of ions, electrons, and reflected and transmitted light, all with thickness, laser focus, and pulse contrast variations. In particular we discuss optimal thickness vs. contrast and details of ultra-thin target normal ion acceleration, along with supporting particle-in-cell studies. This work was supported by the DARPA PULSE program through AMRDEC, by the NNSA (DE-NA0001976), by EC Horizon 2020 LASERLAB-EUROPE/LEPP (654148), and by the German Federal Ministry of Education and Research (BMBF, 03Z1O511).

  18. INTERACTION OF LASER RADIATION WITH MATTER AND OTHER LASER APPLICATIONS: Changes in the emission properties of metal targets during pulse-periodic laser irradiation

    NASA Astrophysics Data System (ADS)

    Konov, Vitalii I.; Pimenov, S. M.; Prokhorov, A. M.; Chapliev, N. I.

    1988-02-01

    A scanning electron microscope was used with a pulse-periodic CO2 laser to discover the laws governing the correlation of the modified microrelief of metal surfaces, subjected to the action of multiple laser pulses, with the emission of charged particles and the luminescence of the irradiated zone. It was established that the influence of sorption and laser-induced desorption on the emission signals may be manifested differently depending on the regime of current generation in the "target-vacuum chamber" circuit.

  19. Electro-optic analysis of the influence of target geometry on electromagnetic pulses generated by petawatt laser-matter interactions

    NASA Astrophysics Data System (ADS)

    Robinson, Timothy; Giltrap, Samuel; Eardley, Samuel; Consoli, Fabrizio; De Angelis, Riccardo; Ingenito, Francesco; Stuart, Nicholas; Verona, Claudio; Smith, Roland A.

    2018-01-01

    We present an analysis of strong laser-driven electromagnetic pulses using novel electro-optic diagnostic techniques. A range of targets were considered, including thin plastic foils (20-550 nm) and mass-limited, optically-levitated micro-targets. Results from foils indicate a dependence of EMP on target thickness, with larger peak electric fields observed with thinner targets. Spectral analysis suggests high repeatability between shots, with identified spectral features consistently detected with <1 MHz standard deviations of the peak position. This deviation is reduced for shots taken on the same day, suggesting that local conditions, such as movement of metal objects within the target chamber, are more likely to lead to minor spectral modifications, highlighting the role of the local environment in determining the details of EMP production. Levitated targets are electrically isolated from their environment, hence these targets should be unable to draw a neutralization current from the earth following ejection of hot electrons from the plasma, in contrast to predictions for pin-mounted foils in the Poyé EMP generation model. With levitated targets, no EMP was measurable above the noise threshold of any diagnostic, despite observation of protons accelerated to >30 MeV energies, suggesting the discharge current contribution to EMP is dominant.

  20. Purification-Free, Target-Selective Immobilization of a Protein from Cell Lysates.

    PubMed

    Cha, Jaehyun; Kwon, Inchan

    2018-02-27

    Protein immobilization has been widely used for laboratory experiments and industrial processes. Preparation of a recombinant protein for immobilization usually requires laborious and expensive purification steps. Here, a novel purification-free, target-selective immobilization technique of a protein from cell lysates is reported. Purification steps are skipped by immobilizing a target protein containing a clickable non-natural amino acid (p-azidophenylalanine) in cell lysates onto alkyne-functionalized solid supports via bioorthogonal azide-alkyne cycloaddition. In order to achieve a target protein-selective immobilization, p-azidophenylalanine was introduced into an exogenous target protein, but not into endogenous non-target proteins using host cells with amber codon-free genomic DNAs. Immobilization of superfolder fluorescent protein (sfGFP) from cell lysates is as efficient as that of the purified sfGFP. Using two fluorescent proteins (sfGFP and mCherry), the authors also demonstrated that the target proteins are immobilized with a minimal immobilization of non-target proteins (target-selective immobilization). © 2018 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  1. Novel target design for enhanced laser driven proton acceleration

    NASA Astrophysics Data System (ADS)

    Dalui, Malay; Kundu, M.; Tata, Sheroy; Lad, Amit D.; Jha, J.; Ray, Krishanu; Krishnamurthy, M.

    2017-09-01

    We demonstrate a simple method of preparing structured target for enhanced laser-driven proton acceleration under target-normal-sheath-acceleration scheme. A few layers of genetically modified, clinically grown micron sized E. Coli bacteria cell coated on a thin metal foil has resulted in an increase in the maximum proton energy by about 1.5 times and the total proton yield is enhanced by approximately 25 times compared to an unstructured reference foil at a laser intensity of 1019 W/cm2. Particle-in-cell simulations on the system shows that the structures on the target-foil facilitates anharmonic resonance, contributing to enhanced hot electron production which leads to stronger accelerating field. The effect is observed to grow as the number of structures is increased in the focal area of the laser pulse.

  2. Visualisation of an nsPEF induced calcium wave using the genetically encoded calcium indicator GCaMP in U87 human glioblastoma cells.

    PubMed

    Carr, Lynn; Bardet, Sylvia M; Arnaud-Cormos, Delia; Leveque, Philippe; O'Connor, Rodney P

    2018-02-01

    Cytosolic, synthetic chemical calcium indicators are typically used to visualise the rapid increase in intracellular calcium ion concentration that follows nanosecond pulsed electric field (nsPEF) application. This study looks at the application of genetically encoded calcium indicators (GECIs) to investigate the spatiotemporal nature of nsPEF-induced calcium signals using fluorescent live cell imaging. Calcium responses to 44kV/cm, 10ns pulses were observed in U87-MG cells expressing either a plasma membrane targeted GECI (GCaMP5-G), or one cytosolically expressed (GCaMP6-S), and compared to the response of cells loaded with cytosolic or plasma membrane targeted chemical calcium indicators. Application of 100 pulses, to cells containing plasma membrane targeted indicators, revealed a wave of calcium across the cell initiating at the cathode side. A similar spatial wave was not observed with cytosolic indicators with mobile calcium buffering properties. The speed of the wave was related to pulse application frequency and it was not propagated by calcium induced calcium release. Copyright © 2017 Elsevier B.V. All rights reserved.

  3. Release of Cell-free MicroRNA Tumor Biomarkers into the Blood Circulation with Pulsed Focused Ultrasound: A Noninvasive, Anatomically Localized, Molecular Liquid Biopsy

    PubMed Central

    Chevillet, John R.; Khokhlova, Tatiana D.; Giraldez, Maria D.; Schade, George R.; Starr, Frank; Wang, Yak-Nam; Gallichotte, Emily N.; Wang, Kai; Hwang, Joo Ha

    2017-01-01

    Purpose To compare the abilities of three pulsed focused ultrasound regimes (that cause tissue liquefaction, permeabilization, or mild heating) to release tumor-derived microRNA into the circulation in vivo and to evaluate release dynamics. Materials and Methods All rat experiments were approved by the University of Washington Institutional Animal Care and Use Committee. Reverse-transcription quantitative polymerase chain reaction array profiling was used to identify candidate microRNA biomarkers in a rat solid tumor cell line. Rats subcutaneously grafted with these cells were randomly assigned among three pulsed focused ultrasound treatment groups: (a) local tissue liquefaction via boiling histotripsy, (b) tissue permeabilization via inertial cavitation, and (c) mild (<10°C) heating of tissue, as well as a sham-treated control group. Blood specimens were drawn immediately prior to treatment and serially over 24 hours afterward. Plasma microRNA was quantified with reverse-transcription quantitative polymerase chain reaction, and statistical significance was determined with one-way analysis of variance (Kruskal-Wallis and Friedman tests), followed by the Dunn multiple-comparisons test. Results After tissue liquefaction and cavitation treatments (but not mild heating), plasma quantities of candidate biomarkers increased significantly (P value range, <.0001 to .04) relative to sham-treated controls. A threefold to 32-fold increase occurred within 15 minutes after initiation of pulsed focused ultrasound tumor treatment, and these increases persisted for 3 hours. Histologic examination confirmed complete liquefaction of the targeted tumor area with boiling histotripsy, in addition to areas of petechial hemorrhage and tissue disruption by means of cavitation-based treatment. Conclusion Mechanical tumor tissue disruption with pulsed focused ultrasound–induced bubble activity significantly increases the plasma abundance of tumor-derived microRNA rapidly after treatment

  4. Embedded 32-bit Differential Pulse Voltammetry (DPV) Technique for 3-electrode Cell Sensing

    NASA Astrophysics Data System (ADS)

    N, Aqmar N. Z.; Abdullah, W. F. H.; Zain, Z. M.; Rani, S.

    2018-03-01

    This paper addresses the development of differential pulse voltammetry (DPV) embedded algorithm using an ARM cortex processor with new developed potentiostat circuit design for in-situ 3-electrode cell sensing. This project is mainly to design a low cost potentiostat for the researchers in laboratories. It is required to develop an embedded algorithm for analytical technique to be used with the designed potentiostat. DPV is one of the most familiar pulse technique method used with 3-electrode cell sensing in chemical studies. Experiment was conducted on 10mM solution of Ferricyanide using the designed potentiostat and the developed DPV algorithm. As a result, the device can generate an excitation signal of DPV from 0.4V to 1.2V and produced a peaked voltammogram with relatively small error compared to the commercial potentiostat; which is only 6.25% difference in peak potential reading. The design of potentiostat device and its DPV algorithm is verified.

  5. Optically pulsed electron accelerator

    DOEpatents

    Fraser, John S.; Sheffield, Richard L.

    1987-01-01

    An optically pulsed electron accelerator can be used as an injector for a free electron laser and comprises a pulsed light source, such as a laser, for providing discrete incident light pulses. A photoemissive electron source emits electron bursts having the same duration as the incident light pulses when impinged upon by same. The photoemissive electron source is located on an inside wall of a radio frequency powered accelerator cell which accelerates the electron burst emitted by the photoemissive electron source.

  6. Optically pulsed electron accelerator

    DOEpatents

    Fraser, J.S.; Sheffield, R.L.

    1985-05-20

    An optically pulsed electron accelerator can be used as an injector for a free electron laser and comprises a pulsed light source, such as a laser, for providing discrete incident light pulses. A photoemissive electron source emits electron bursts having the same duration as the incident light pulses when impinged upon by same. The photoemissive electron source is located on an inside wall of a radiofrequency-powered accelerator cell which accelerates the electron burst emitted by the photoemissive electron source.

  7. Ion mediated targeting of cells with nanoparticles

    NASA Astrophysics Data System (ADS)

    Maheshwari, Vivek; Fu, Jinlong

    2010-03-01

    In eukaryotic cells, Ca^2+ ions are necessary for intracellular signaling, in activity of mitochondria and a variety of other cellular process that have been linked to cell apoptosis, proteins synthesis and cell-cycle regulation. Here we show that Ca^2+ ions, serving as the bio-compatible interface can be used to target Saccharomyces cerevisiae (SaC, baker's yeast), a model eukaryotic cell, with Au nanoparticles (10 nm). The Ca^2+ ions bind to the carboxylic acid groups in the citrate functionalized Au nanoparticles. This transforms the nanoparticles into micron long 1-D branched chain assemblies due to inter-particle dipole-dipole interaction and inter-particle bonding due to the divalent nature of the Ca^2+ ion. A similar transformation is observed with the use of divalent ions Mg^2+, Cd^2+ and Fe^2+. The 1-D assembly aids the interfacing of ion-nanoparticles on the cell by providing multiple contact points. Further monovalent ions such as Na^+ are also effective for the targeting of the cell with nanoparticles. However Na-Au nanoparticles are limited in their deposition as they exist in solution as single particles. The cells remain alive after the deposition process and their vitality is unaffected by the interfacing with ion-nanoparticles.

  8. T-REX on-demand redox targeting in live cells.

    PubMed

    Parvez, Saba; Long, Marcus J C; Lin, Hong-Yu; Zhao, Yi; Haegele, Joseph A; Pham, Vanha N; Lee, Dustin K; Aye, Yimon

    2016-12-01

    This protocol describes targetable reactive electrophiles and oxidants (T-REX)-a live-cell-based tool designed to (i) interrogate the consequences of specific and time-resolved redox events, and (ii) screen for bona fide redox-sensor targets. A small-molecule toolset comprising photocaged precursors to specific reactive redox signals is constructed such that these inert precursors specifically and irreversibly tag any HaloTag-fused protein of interest (POI) in mammalian and Escherichia coli cells. Syntheses of the alkyne-functionalized endogenous reactive signal 4-hydroxynonenal (HNE(alkyne)) and the HaloTag-targetable photocaged precursor to HNE(alkyne) (also known as Ht-PreHNE or HtPHA) are described. Low-energy light prompts photo-uncaging (t 1/2 <1-2 min) and target-specific modification. The targeted modification of the POI enables precisely timed and spatially controlled redox events with no off-target modification. Two independent pathways are described, along with a simple setup to functionally validate known targets or discover novel sensors. T-REX sidesteps mixed responses caused by uncontrolled whole-cell swamping with reactive signals. Modification and downstream response can be analyzed by in-gel fluorescence, proteomics, qRT-PCR, immunofluorescence, fluorescence resonance energy transfer (FRET)-based and dual-luciferase reporters, or flow cytometry assays. T-REX targeting takes 4 h from initial probe treatment. Analysis of targeted redox responses takes an additional 4-24 h, depending on the nature of the pathway and the type of readouts used.

  9. T-REX on-demand redox targeting in live cells

    PubMed Central

    Parvez, Saba; Long, Marcus J C; Lin, Hong-Yu; Zhao, Yi; Haegele, Joseph A; Pham, Vanha N; Lee, Dustin K; Aye, Yimon

    2017-01-01

    This protocol describes targetable reactive electrophiles and oxidants (T-REX)—a live-cell-based tool designed to (i) interrogate the consequences of specific and time-resolved redox events, and (ii) screen for bona fide redox-sensor targets. A small-molecule toolset comprising photocaged precursors to specific reactive redox signals is constructed such that these inert precursors specifically and irreversibly tag any HaloTag-fused protein of interest (POI) in mammalian and Escherichia coli cells. Syntheses of the alkyne-functionalized endogenous reactive signal 4-hydroxynonenal (HNE (alkyne)) and the HaloTag-targetable photocaged precursor to HNE (alkyne) (also known as Ht-PreHNE or HtPHA) are described. Low-energy light prompts photo-uncaging (t1/2 <1–2 min) and target-specific modification. The targeted modification of the POI enables precisely timed and spatially controlled redox events with no off-target modification. Two independent pathways are described, along with a simple setup to functionally validate known targets or discover novel sensors. T-REX sidesteps mixed responses caused by uncontrolled whole-cell swamping with reactive signals. Modification and downstream response can be analyzed by in-gel fluorescence, proteomics, qRT-PCR, immunofluorescence, fluorescence resonance energy transfer (FRET)-based and dual-luciferase reporters, or flow cytometry assays. T-REX targeting takes 4 h from initial probe treatment. Analysis of targeted redox responses takes an additional 4–24 h, depending on the nature of the pathway and the type of readouts used. PMID:27809314

  10. Particle in cell simulation on plasma grating contrast enhancement induced by infrared laser pulse

    NASA Astrophysics Data System (ADS)

    Li, M.; Yuan, T.; Xu, Y. X.; Wang, J. X.; Luo, S. N.

    2018-05-01

    The dynamics of plasma grating contrast enhancement (PGCE) irradiated by an infrared laser pulse is investigated with one dimensional particle-in-cell simulation where field ionization and impact ionization are simultaneously considered for the first time. The numeric results show that the impact ionization dominates the PGCE process. Upon the interaction with the laser pulse, abundant free electrons are efficiently accelerated and subsequently triggered massive impact ionizations in the density ridges of the plasma grating for the higher local plasma energy density, which efficiently enhances the grating contrast. Besides the dynamic analysis of PGCE, we explore the parameter space of the incident infrared laser pulse to optimize the PGCE effect, which can provide useful guidance to experiments related to laser-plasma-grating interactions and may find applications in prolonging the duration of the plasma grating.

  11. Landscape phages and their fusion proteins targeted to breast cancer cells

    PubMed Central

    Fagbohun, Olusegun A.; Bedi, Deepa; Grabchenko, Natalia I.; Deinnocentes, Patricia A.; Bird, Richard C.; Petrenko, Valery A.

    2012-01-01

    Breast cancer is a leading cause of death among women in the USA. The efficacy of existing anticancer therapeutics can be improved by targeting them through conjugation with ligands binding to cellular receptors. Recently, we developed a novel drug targeting strategy based on the use of pre-selected cancer-specific ‘fusion pVIII proteins’ (fpVIII), as targeting ligands. To study the efficiency of this approach in animal models, we developed a panel of breast cancer cell-binding phages as a source of targeted fpVIIIs. Two landscape phage peptide libraries (8-mer f8/8 and 9-mer f8/9) were screened to isolate 132 phage variants that recognize breast carcinoma cells MCF-7 and ZR-75-1 and internalize into the cells. When tested for their interaction with the breast cancer cells in comparison with liver cancer cells HepG2, human mammary cells MCF-10A cells and serum, 16 of the phage probes selectively interacted with the breast cancer cells whereas 32 bound both breast and liver cancer cells. The most prominent cancer-specific phage DMPGTVLP, demonstrating sub-nanomolar Kd in interaction with target cells, was used for affinity chromatography of cellular membrane molecules to reveal its potential binding receptor. The isolated protein was identified by direct sequencing as cellular surface nucleolin. This conclusion was confirmed by inhibition of the phage–cell interaction with nucleolin antibodies. Other prominent phage binders VPTDTDYS, VEEGGYIAA, and DWRGDSMDS demonstrate consensus motifs common to previously identified cancer-specific peptides. Isolated phage proteins exhibit inherent binding specificity towards cancer cells, demonstrating the functional activity of the selected fused peptides. The selected phages, their peptide inserts and intact fusion proteins can serve as promising ligands for the development of targeted nanomedicines and their study in model mice with xenograft of human cells MCF-7 and ZR-75-1. PMID:22490956

  12. Modular cell-internalizing aptamer nanostructure enables targeted delivery of large functional RNAs in cancer cell lines.

    PubMed

    Porciani, David; Cardwell, Leah N; Tawiah, Kwaku D; Alam, Khalid K; Lange, Margaret J; Daniels, Mark A; Burke, Donald H

    2018-06-11

    Large RNAs and ribonucleoprotein complexes have powerful therapeutic potential, but effective cell-targeted delivery tools are limited. Aptamers that internalize into target cells can deliver siRNAs (<15 kDa, 19-21 nt/strand). We demonstrate a modular nanostructure for cellular delivery of large, functional RNA payloads (50-80 kDa, 175-250 nt) by aptamers that recognize multiple human B cell cancer lines and transferrin receptor-expressing cells. Fluorogenic RNA reporter payloads enable accelerated testing of platform designs and rapid evaluation of assembly and internalization. Modularity is demonstrated by swapping in different targeting and payload aptamers. Both modules internalize into leukemic B cell lines and remained colocalized within endosomes. Fluorescence from internalized RNA persists for ≥2 h, suggesting a sizable window for aptamer payloads to exert influence upon targeted cells. This demonstration of aptamer-mediated, cell-internalizing delivery of large RNAs with retention of functional structure raises the possibility of manipulating endosomes and cells by delivering large aptamers and regulatory RNAs.

  13. ELECTRIC PULSE GENERATOR

    DOEpatents

    Buntenbach, R.W.

    1959-06-01

    S>An electro-optical apparatus is described which produces electric pulses in programmed sequences at times and durations controlled with great accuracy. An oscilloscope CRT is supplied with signals to produce a luminous spot moving in a circle. An opaque mask with slots of variable width transmits light from the spot to a photoelectric transducer. For shorter pulse decay times a CRT screen which emits UV can be used with a UVtransmitting filter and a UV- sensitive photoelectric cell. Pulses are varied by changing masks or by using masks with variable slots. This device may be used in multiple arrangements to produce other pulse aT rangements, or it can be used to trigger an electronic pulse generator. (T.R.H.)

  14. Asymmetrical Transmembrane Potential in Intracellular Organelles of Adrenal Chromatin Cells Exposed to Nanosecond Electric Pulses

    NASA Astrophysics Data System (ADS)

    Aramendia Zabaleta, Guillermo Jose

    In our research on exploring the effects of 5 ns, high intensity electric pulses on neurosecretory adrenal chromaffin cells, cell modeling has played an important role in understanding and explaining the experimental results. Externally applied nanosecond-duration electric pulses (NEPs) can affect cells by creating nanopores in the cell and intracellular organelle membranes, making these membranes permeable to certain ions. A chromaffin cell contains, at a minimum, 7000 secretory granules plus other organelles such as mitochondria and the endoplasmic reticulum. In all the biological cell models constructed in the literature, there is no evidence of asymmetrical Transmembrane Potential (TMP) distribution in the intracellular membranes. However, these models do not include a realistic number of intracellular organelles. The goal of this research was to construct a more realistic cell model that incorporates a large number of secretory granules in the cytosol. To this end, a beta-version of the real-valued unstructured mesh Finite Element Method (FEM) electro-quasi-static module in Sim4life (SPEAG, Switzerland) has been used to model a chromaffin cell in which 1000 secretory granules are included in the cytosol. The model is, we believe, the most detailed geometrical cell model developed. It includes a spherical chromaffin cell (radius 8 mum), nucleus (radius 2.5 mum) located off-center, 500 granules (radius 200 nm) randomly located within a distance of 2 mum from the surface of the nucleus, and additional 500 granules randomly located in the remaining region of the cytosol. Cell and granule membrane thickness was set to 5 nm and nuclear membrane thickness to 10 nm. Dielectric properties of all constituents of the model were obtained from the literature or measured. Because the FEM Low Frequency solver is a quasi-static solver and not capable of accepting a time-varying pulse as input, all computations have been performed at single frequencies in the range DC to 60

  15. Label-free resistive-pulse cytometry.

    PubMed

    Chapman, M R; Sohn, L L

    2011-01-01

    Numerous methods have recently been developed to characterize cells for size, shape, and specific cell-surface markers. Most of these methods rely upon exogenous labeling of the cells and are better suited for large cell populations (>10,000). Here, we review a label-free method of characterizing and screening cells based on the Coulter-counter technique of particle sizing: an individual cell transiting a microchannel (or "pore") causes a downward pulse in the measured DC current across that "pore". Pulse magnitude corresponds to the cell size, pulse width to the transit time needed for the cell to pass through the pore, and pulse shape to how the cell traverses across the pore (i.e., rolling or tumbling). When the pore is functionalized with an antibody that is specific to a surface-epitope of interest, label-free screening of a specific marker is possible, as transient binding between the two results in longer time duration than when the pore is unfunctionalized or functionalized with a nonspecific antibody. While this method cannot currently compete with traditional technology in terms of throughput, there are a number of applications for which this technology is better suited than current commercial cytometry systems. Applications include the rapid and nondestructive analysis of small cell populations (<100), which is not possible with current technology, and a platform for providing true point-of-care clinical diagnostics, due to the simplicity of the device, low manufacturing costs, and ease of use. Copyright © 2011 Elsevier Inc. All rights reserved.

  16. Dynamic effects and applications for nanosecond pulsed electric fields in cells and tissues

    NASA Astrophysics Data System (ADS)

    Beebe, Stephen J.; Blackmore, Peter F.; Hall, Emily; White, Jody A.; Willis, Lauren K.; Fauntleroy, Laura; Kolb, Juergen F.; Schoenbach, Karl H.

    2005-04-01

    Nanosecond, high intensity pulsed electric fields [nsPEFs] that are below the plasma membrane [PM] charging time constant have decreasing effects on the PM and increasing effects on intracellular structures and functions as the pulse duration decreases. When human cell suspensions were exposed to nsPEFs where the electric fields were sufficiently intense [10-300ns, <=300 kV/cm.], apoptosis signaling pathways could be activated in several cell models. Multiple apoptosis markers were observed in Jurkat, HL-60, 3T3L1-preadipocytes, and isolated rat adipocytes including decreased cell size and number, caspase activation, DNA fragmentation, and/or cytochrome c release into the cytoplasm. Phosphatidylserine externalization was observed as a biological response to nsPEFs in 3T3-L1 preadipocytes and p53-wildtype and -null human colon carcinoma cells. B10.2 mouse fibrosarcoma tumors that were exposed to nsPEFs ex vivo and in vivo exhibited DNA fragmentation, elevated caspase activity, and reduced size and weight compared to contralateral sham-treated control tumors. When nsPEF conditions were below thresholds for apoptosis and classical PM electroporation, non-apoptotic responses were observed similar to those initiated through PM purinergic receptors in HL-60 cells and thrombin in human platelets. These included Ca2+ mobilization from intracellular stores [endoplasmic reticulum] and subsequently through store-operated Ca2+ channels in the PM. In addition, platelet activation measured as aggregation responses were observed in human platelets. Finally, when nsPEF conditions followed classical electroporation-mediated transfection, the expression intensity and number of GFP-expressing cells were enhanced above cells exposed to electroporation conditions alone. These studies demonstrate that application of nsPEFs to cells or tissues can modulate cell-signaling mechanisms with possible applications as a new basic science tool, cancer treatment, wound healing, and gene therapy.

  17. The cancer cell adhesion resistome: mechanisms, targeting and translational approaches.

    PubMed

    Dickreuter, Ellen; Cordes, Nils

    2017-06-27

    Cell adhesion-mediated resistance limits the success of cancer therapies and is a great obstacle to overcome in the clinic. Since the 1990s, where it became clear that adhesion of tumor cells to the extracellular matrix is an important mediator of therapy resistance, a lot of work has been conducted to understand the fundamental underlying mechanisms and two paradigms were deduced: cell adhesion-mediated radioresistance (CAM-RR) and cell adhesion-mediated drug resistance (CAM-DR). Preclinical work has evidently demonstrated that targeting of integrins, adapter proteins and associated kinases comprising the cell adhesion resistome is a promising strategy to sensitize cancer cells to both radiotherapy and chemotherapy. Moreover, the cell adhesion resistome fundamentally contributes to adaptation mechanisms induced by radiochemotherapy as well as molecular drugs to secure a balanced homeostasis of cancer cells for survival and growth. Intriguingly, this phenomenon provides a basis for synthetic lethal targeted therapies simultaneously administered to standard radiochemotherapy. In this review, we summarize current knowledge about the cell adhesion resistome and highlight targeting strategies to override CAM-RR and CAM-DR.

  18. Application of stem cells in targeted therapy of breast cancer: a systematic review.

    PubMed

    Madjd, Zahra; Gheytanchi, Elmira; Erfani, Elham; Asadi-Lari, Mohsen

    2013-01-01

    The aim of this systematic review was to investigate whether stem cells could be effectively applied in targeted therapy of breast cancer. A systematic literature search was performed for original articles published from January 2007 until May 2012. Nine studies met the inclusion criteria for phase I or II clinical trials, of which three used stem cells as vehicles, two trials used autologous hematopoetic stem cells and in four trials cancer stem cells were targeted. Mesenchymal stem cells (MSCs) were applied as cellular vehicles to transfer therapeutic agents. Cell therapy with MSC can successfully target resistant cancers. Cancer stem cells were selectively targeted via a proteasome-dependent suicide gene leading to tumor regression. Wnt/β-catenin signaling pathway has been also evidenced to be an attractive CSC-target. This systematic review focused on two different concepts of stem cells and breast cancer marking a turning point in the trials that applied stem cells as cellular vehicles for targeted delivery therapy as well as CSC-targeted therapies. Applying stem cells as targeted therapy could be an effective therapeutic approach for treatment of breast cancer in the clinic and in therapeutic marketing; however this needs to be confirmed with further clinical investigations.

  19. High Energy electron and proton acceleration by circularly polarized laser pulse from near critical density hydrogen gas target.

    PubMed

    Sharma, Ashutosh

    2018-02-01

    Relativistic electron rings hold the possibility of very high accelerating rates, and hopefully a relatively cheap and compact accelerator/collimator for ultrahigh energy proton source. In this work, we investigate the generation of helical shaped quasi-monoenergetic relativistic electron beam and high-energy proton beam from near critical density plasmas driven by petawatt-circularly polarized-short laser pulses. We numerically observe the efficient proton acceleration from magnetic vortex acceleration mechanism by using the three dimensional particle-in-cell simulations; proton beam with peak energy 350 MeV, charge ~10nC and conversion efficiency more than 6% (which implies 2.4 J proton beam out of the 40 J incident laser energy) is reported. We detailed the microphysics involved in the ion acceleration mechanism, which requires investigating the role of self-generated plasma electric and magnetic fields. The concept of efficient generation of quasi-monoenergetic electron and proton beam from near critical density gas targets may be verified experimentally at advanced high power - high repetition rate laser facilities e.g. ELI-ALPS. Such study should be an important step towards the development of high quality electron and proton beam.

  20. Alternating-pulse iontophoresis for targeted cutaneous anesthesia

    NASA Technical Reports Server (NTRS)

    Meyer, Peter F.; Oddsson, Lars I E.

    2003-01-01

    In studies of sensory contributions to motor control, it may be advantageous to temporarily reduce the sensitivity of specific sensory systems. This article details a method for non-invasively inducing cutaneous anesthesia, leaving proprioceptive and motor functions intact. This method, called alternating-pulse iontophoresis, differs from conventional direct-current (DC) iontophoretic drug delivery in that adjacent drug delivery electrodes are stimulated out-of-phase. The total current delivered at any instant is then less than that produced during a comparable DC application, while the uniformity of drug delivery is expected to improve. Effective delivery of local anesthetics to the cutaneous foot soles by alternating-pulse iontophoresis was demonstrated using cutaneous pressure sensory threshold levels (STL's) assessed with Semmes-Weinstein monofilaments (arbitrary units of perceived force, or a.u.). Thirteen of 16 healthy subjects achieved a level of anesthesia greater than or equal to that normally associated with clinical peripheral sensory neuropathy. Average STL's measured prior to the anesthesia procedure were 4.00 a.u. ( approximately 10 mN). Immediately following the procedure, STL's were elevated to an average of 5.40 a.u. ( approximately 246 mN) and averaged 4.97 a.u. ( approximately 92 mN) after 50 min of standing. A number of research and clinical applications for this technique are suggested.

  1. Method and apparatus for pulse stacking

    DOEpatents

    Harney, Robert C.

    1977-01-01

    An active pulse stacking system including an etalon and an electro-optical modulator apparatus combined with a pulse-forming network capable of forming and summing a sequence of time-delayed optical waveforms arising from, for example, a single laser pulse. The Pockels cell pulse stacker may attain an efficiency of about 2.6% while providing a controllable faster-than-exponential time rise in transmitted pulse intensity.

  2. Intense isolated attosecond pulse generation from relativistic laser plasmas using few-cycle laser pulses

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    Ma, Guangjin, E-mail: guangjin.ma@mpq.mpg.de; Max-Planck-Institut für Quantenoptik, D-85748 Garching; Dallari, William

    2015-03-15

    We have performed a systematic study through particle-in-cell simulations to investigate the generation of attosecond pulse from relativistic laser plasmas when laser pulse duration approaches the few-cycle regime. A significant enhancement of attosecond pulse energy has been found to depend on laser pulse duration, carrier envelope phase, and plasma scale length. Based on the results obtained in this work, the potential of attaining isolated attosecond pulses with ∼100 μJ energy for photons >16 eV using state-of-the-art laser technology appears to be within reach.

  3. Agricultural and Food Processing Applications of Pulsed Power and Plasma Technologies

    NASA Astrophysics Data System (ADS)

    Takaki, Koichi

    Agricultural and food processing applications of pulsed power and plasma technologies are described in this paper. Repetitively operated compact pulsed power generators with a moderate peak power are developed for the agricultural and the food processing applications. These applications are mainly based on biological effects and can be categorized as germination control of plants such as Basidiomycota and arabidopsis inactivation of bacteria in soil and liquid medium of hydroponics; extraction of juice from fruits and vegetables; decontamination of air and liquid, etc. Types of pulsed power that have biological effects are caused with gas discharges, water discharges, and electromagnetic fields. The discharges yield free radicals, UV radiation, intense electric field, and shock waves. Biologically based applications of pulsed power and plasma are performed by selecting the type that gives the target objects the adequate result from among these agents or byproducts. For instance, intense electric fields form pores on the cell membrane, which is called electroporation, or influence the nuclei. This paper mainly describes the application of the pulsed power for the germination control of Basidiomycota i.e. mushroom, inactivation of fungi in the soil and the liquid medium in hydroponics, and extraction of polyphenol from skins of grape.

  4. Concise Review: Emerging Drugs Targeting Epithelial Cancer Stem-Like Cells.

    PubMed

    Ahmed, Mehreen; Chaudhari, Kritika; Babaei-Jadidi, Roya; Dekker, Lodewijk V; Shams Nateri, Abdolrahman

    2017-04-01

    Increasing evidence suggests that cancer cell populations contain a small proportion of cells that display stem-like cell properties and which may be responsible for overall tumor maintenance. These cancer stem-like cells (CSCs) appear to have unique tumor-initiating ability and innate survival mechanisms that allow them to resist cancer therapies, consequently promoting relapses. Selective targeting of CSCs may provide therapeutic benefit and several recent reports have indicated this may be possible. In this article, we review drugs targeting CSCs, in selected epithelial cell-derived cancers. Stem Cells 2017;35:839-850. © 2017 AlphaMed Press.

  5. MicroRNA-944 Affects Cell Growth by Targeting EPHA7 in Non-Small Cell Lung Cancer.

    PubMed

    Liu, Minxia; Zhou, Kecheng; Cao, Yi

    2016-09-26

    MicroRNAs (miRNAs) have critical roles in lung tumorigenesis and development. To determine aberrantly expressed miRNAs involved in non-small cell lung cancer (NSCLC) and investigate pathophysiological functions and mechanisms, we firstly carried out small RNA deep sequencing in NSCLC cell lines (EPLC-32M1, A549 and 801D) and a human immortalized cell line 16HBE, we then studied miRNA function by cell proliferation and apoptosis. cDNA microarray, luciferase reporter assay and miRNA transfection were used to investigate interaction between the miRNA and target gene. miR-944 was significantly down-regulated in NSCLC and had many putative targets. Moreover, the forced expression of miR-944 significantly inhibited the proliferation of NSCLC cells in vitro. By integrating mRNA expression data and miR-944-target prediction, we disclosed that EPHA7 was a potential target of miR-944, which was further verified by luciferase reporter assay and microRNA transfection. Our data indicated that miR-944 targets EPHA7 in NSCLC and regulates NSCLC cell proliferation, which may offer a new mechanism underlying the development and progression of NSCLC.

  6. The effect of pulsed electric fields on the electrotactic migration of human neural progenitor cells through the involvement of intracellular calcium signaling.

    PubMed

    Hayashi, Hisamitsu; Edin, Fredrik; Li, Hao; Liu, Wei; Rask-Andersen, Helge

    2016-12-01

    Endogenous electric fields (EFs) are required for the physiological control of the central nervous system development. Application of the direct current EFs to neural stem cells has been studied for the possibility of stem cell transplantation as one of the therapies for brain injury. EFs generated within the nervous system are often associated with action potentials and synaptic activity, apparently resulting in a pulsed current in nature. The aim of this study is to investigate the effect of pulsed EF, which can reduce the cytotoxicity, on the migration of human neural progenitor cells (hNPCs). We applied the mono-directional pulsed EF with a strength of 250mV/mm to hNPCs for 6h. The migration distance of the hNPCs exposed to pulsed EF was significantly greater compared with the control not exposed to the EF. Pulsed EFs, however, had less of an effect on the migration of the differentiated hNPCs. There was no significant change in the survival of hNPCs after exposure to the pulsed EF. To investigate the role of Ca 2+ signaling in electrotactic migration of hNPCs, pharmacological inhibition of Ca 2+ channels in the EF-exposed cells revealed that the electrotactic migration of hNPCs exposed to Ca 2+ channel blockers was significantly lower compared to the control group. The findings suggest that the pulsed EF induced migration of hNPCs is partly influenced by intracellular Ca 2+ signaling. Copyright © 2016 Elsevier B.V. All rights reserved.

  7. New Strategies in Engineering T-cell Receptor Gene-Modified T cells to More Effectively Target Malignancies.

    PubMed

    Schmitt, Thomas M; Stromnes, Ingunn M; Chapuis, Aude G; Greenberg, Philip D

    2015-12-01

    The immune system, T cells in particular, have the ability to target and destroy malignant cells. However, antitumor immune responses induced from the endogenous T-cell repertoire are often insufficient for the eradication of established tumors, as illustrated by the failure of cancer vaccination strategies or checkpoint blockade for most tumors. Genetic modification of T cells to express a defined T-cell receptor (TCR) can provide the means to rapidly generate large numbers of tumor-reactive T cells capable of targeting tumor cells in vivo. However, cell-intrinsic factors as well as immunosuppressive factors in the tumor microenvironment can limit the function of such gene-modified T cells. New strategies currently being developed are refining and enhancing this approach, resulting in cellular therapies that more effectively target tumors and that are less susceptible to tumor immune evasion. ©2015 American Association for Cancer Research.

  8. Pulsed-DC selfsputtering of copper

    NASA Astrophysics Data System (ADS)

    Wiatrowski, A.; Posadowski, W. M.; Radzimski, Z. J.

    2008-03-01

    At standard magnetron sputtering conditions (argon pressure ~0.5 Pa) inert gas particles are often entrapped in the formed films. Inert gas contamination can be eliminated by using the self-sustained magnetron sputtering process because it is done in the absence of the inert gas atmosphere. The self-sustained sputtering (SSS) gives also a unique condition during the transport of sputtered particles to the substrate. It is especially useful for filling high aspect ratio submicron scale structures for microelectronics. So far it has been shown that the self-sputtering process can be sustained in the DC operation mode (DC-SSS) only. The main disadvantage of DC-SSS process is instability related to possible arc formation. Usage of pulsed sputtering, similarly to reactive pulsed magnetron sputtering, could eliminate this problem. In this paper results of pulsed-DC self-sustained magnetron sputtering (pulsed DC-SSS) of copper are presented for the first time. The planar magnetron equipped with a 50 mm in diameter and 6 mm thick copper target was powered by DC-power supply modulated by power switch. The maximum target power was about 11 kW (~550W/cm2). The magnetron operation was investigated as a function of pulsing frequency (20-100 kHz) and duty factor (50-90%). The discharge extinction pressure was determined for these conditions. The plasma emission spectra (400-410nm range) and deposition rates were observed for both DC and pulsed DC sustained self-sputtering processes. The presented results illustrate that stable pulsed DC-SSS process can be obtained at pulsing frequency in the range of 60-100 kHz and duty factor of 70-90%.

  9. Probing Xist RNA Structure in Cells Using Targeted Structure-Seq

    PubMed Central

    Rutenberg-Schoenberg, Michael; Simon, Matthew D.

    2015-01-01

    The long non-coding RNA (lncRNA) Xist is a master regulator of X-chromosome inactivation in mammalian cells. Models for how Xist and other lncRNAs function depend on thermodynamically stable secondary and higher-order structures that RNAs can form in the context of a cell. Probing accessible RNA bases can provide data to build models of RNA conformation that provide insight into RNA function, molecular evolution, and modularity. To study the structure of Xist in cells, we built upon recent advances in RNA secondary structure mapping and modeling to develop Targeted Structure-Seq, which combines chemical probing of RNA structure in cells with target-specific massively parallel sequencing. By enriching for signals from the RNA of interest, Targeted Structure-Seq achieves high coverage of the target RNA with relatively few sequencing reads, thus providing a targeted and scalable approach to analyze RNA conformation in cells. We use this approach to probe the full-length Xist lncRNA to develop new models for functional elements within Xist, including the repeat A element in the 5’-end of Xist. This analysis also identified new structural elements in Xist that are evolutionarily conserved, including a new element proximal to the C repeats that is important for Xist function. PMID:26646615

  10. Cellular response to high pulse repetition rate nanosecond pulses varies with fluorescent marker identity

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    Steelman, Zachary A., E-mail: zachary.steelman@duke.edu; Tolstykh, Gleb P.; Beier, Hope T.

    Nanosecond electric pulses (nsEP's) are a well-studied phenomena in biophysics that cause substantial alterations to cellular membrane dynamics, internal biochemistry, and cytoskeletal structure, and induce apoptotic and necrotic cell death. While several studies have attempted to measure the effects of multiple nanosecond pulses, the effect of pulse repetition rate (PRR) has received little attention, especially at frequencies greater than 100 Hz. In this study, uptake of Propidium Iodide, FM 1–43, and YO-PRO-1 fluorescent dyes in CHO-K1 cells was monitored across a wide range of PRRs (5 Hz–500 KHz) using a laser-scanning confocal microscope in order to better understand how high frequency repetition ratesmore » impact induced biophysical changes. We show that frequency trends depend on the identity of the dye under study, which could implicate transmembrane protein channels in the uptake response due to their chemical selectivity. Finally, YO-PRO-1 fluorescence was monitored in the presence of Gadolinium (Gd{sup 3+}), Ruthenium Red, and in calcium-free solution to elucidate a mechanism for its unique frequency trend. - Highlights: • Pulse repetition rate (PRR) is understudied in nanosecond electric pulsing. • 200 V pulses were applied to CHO-K1 cells from 5 Hz to 500 KHz. • Pulsing was repeated using a variety of fluorophores and imaging conditions. • The response is highly dependent on the fluorophore and the imaging conditions. • This may implicate protein channels in the nanoporation response.« less

  11. Cell targeting peptides as smart ligands for targeting of therapeutic or diagnostic agents: a systematic review.

    PubMed

    Mousavizadeh, Ali; Jabbari, Ali; Akrami, Mohammad; Bardania, Hassan

    2017-10-01

    Cell targeting peptides (CTP) are small peptides which have high affinity and specificity to a cell or tissue targets. They are typically identified by using phage display and chemical synthetic peptide library methods. CTPs have attracted considerable attention as a new class of ligands to delivery specifically therapeutic and diagnostic agents, because of the fact they have several advantages including easy synthesis, smaller physical sizes, lower immunogenicity and cytotoxicity and their simple and better conjugation to nano-carriers and therapeutic or diagnostic agents compared to conventional antibodies. In this systematic review, we will focus on the basic concepts concerning the use of cell-targeting peptides (CTPs), following the approaches of selecting them from peptide libraries. We discuss several developed strategies for cell-specific delivery of different cargos by CTPs, which are designed for drug delivery and diagnostic applications. Copyright © 2017 Elsevier B.V. All rights reserved.

  12. Characterization of nanoparticle mediated laser transfection by femtosecond laser pulses for applications in molecular medicine.

    PubMed

    Schomaker, Markus; Heinemann, Dag; Kalies, Stefan; Willenbrock, Saskia; Wagner, Siegfried; Nolte, Ingo; Ripken, Tammo; Murua Escobar, Hugo; Meyer, Heiko; Heisterkamp, Alexander

    2015-02-03

    In molecular medicine, the manipulation of cells is prerequisite to evaluate genes as therapeutic targets or to transfect cells to develop cell therapeutic strategies. To achieve these purposes it is essential that given transfection techniques are capable of handling high cell numbers in reasonable time spans. To fulfill this demand, an alternative nanoparticle mediated laser transfection method is presented herein. The fs-laser excitation of cell-adhered gold nanoparticles evokes localized membrane permeabilization and enables an inflow of extracellular molecules into cells. The parameters for an efficient and gentle cell manipulation are evaluated in detail. Efficiencies of 90% with a cell viability of 93% were achieved for siRNA transfection. The proof for a molecular medical approach is demonstrated by highly efficient knock down of the oncogene HMGA2 in a rapidly proliferating prostate carcinoma in vitro model using siRNA. Additionally, investigations concerning the initial perforation mechanism are conducted. Next to theoretical simulations, the laser induced effects are experimentally investigated by spectrometric and microscopic analysis. The results indicate that near field effects are the initial mechanism of membrane permeabilization. This methodical approach combined with an automated setup, allows a high throughput targeting of several 100,000 cells within seconds, providing an excellent tool for in vitro applications in molecular medicine. NIR fs lasers are characterized by specific advantages when compared to lasers employing longer (ps/ns) pulses in the visible regime. The NIR fs pulses generate low thermal impact while allowing high penetration depths into tissue. Therefore fs lasers could be used for prospective in vivo applications.

  13. Tumor-targeting domains for chimeric antigen receptor T cells.

    PubMed

    Bezverbnaya, Ksenia; Mathews, Ashish; Sidhu, Jesse; Helsen, Christopher W; Bramson, Jonathan L

    2017-01-01

    Immunotherapy with chimeric antigen receptor (CAR) T cells has been advancing steadily in clinical trials. Since the ability of engineered T cells to recognize intended tumor-associated targets is crucial for the therapeutic success, antigen-binding domains play an important role in shaping T-cell responses. Single-chain antibody and T-cell receptor fragments, natural ligands, repeat proteins, combinations of the above and universal tag-specific domains have all been used in the antigen-binding moiety of chimeric receptors. Here we outline the advantages and disadvantages of different domains, discuss the concepts of affinity and specificity, and highlight the recent progress of each targeting strategy.

  14. A simple solution to the problem of effective utilisation of the target material for pulsed laser deposition of thin films

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    Kuzanyan, A S; Kuzanyan, A A; Petrosyan, V A

    The factors determining the efficiency of the target material utilisation for pulsed laser deposition of films are considered. The target volume is calculated, which is evaporated in the ablation process by the focused laser radiation having a rectangular form. The new device is suggested and developed for obtaining thin films by the method of laser deposition, which is specific in the employment of a simple optical system mounted outside a deposition chamber that comprises two lenses and the diaphragm and focuses the laser beam onto a target in the form of a sector-like spot. Thin films of CuO and YBaCuOmore » were deposited with this device. Several deposition cycles revealed that the target material is consumed uniformly from the entire surface of the target. A maximal spread of the target thickness was not greater than ±2% both prior to deposition and after it. The device designed provides a high coefficient of the target material utilisation efficiency. (laser deposition of thin films)« less

  15. Development of bipolar-pulse accelerator for intense pulsed ion beam acceleration

    NASA Astrophysics Data System (ADS)

    Masugata, Katsumi; Shimizu, Yuichro; Fujioka, Yuhki; Kitamura, Iwao; Tanoue, Hisao; Arai, Kazuo

    2004-12-01

    To improve the purity of intense pulsed ion beams, a new type of pulsed ion beam accelerator named "bipolar pulse accelerator" was proposed. To confirm the principle of the accelerator a prototype of the experimental system was developed. The system utilizes By type magnetically insulated acceleration gap and operated with single polar negative pulse. A coaxial gas puff plasma gun was used as an ion source, which was placed inside the grounded anode. Source plasma (nitrogen) of current density ≈25 A/cm2, duration ≈1.5 μs was injected into the acceleration gap by the plasma gun. The ions were successfully accelerated from the grounded anode to the drift tube by applying negative pulse of voltage 240 kV, duration 100 ns to the drift tube. Pulsed ion beam of current density ≈40 A/cm2, duration ≈50 ns was obtained at 41 mm downstream from the anode surface. To evaluate the irradiation effect of the ion beam to solid material, an amorphous silicon thin film of thickness ≈500 nm was used as the target, which was deposited on the glass substrate. The film was found to be poly-crystallized after 4-shots of the pulsed nitrogen ion beam irradiation.

  16. Optical cell monitoring system for underwater targets

    NASA Astrophysics Data System (ADS)

    Moon, SangJun; Manzur, Fahim; Manzur, Tariq; Demirci, Utkan

    2008-10-01

    We demonstrate a cell based detection system that could be used for monitoring an underwater target volume and environment using a microfluidic chip and charge-coupled-device (CCD). This technique allows us to capture specific cells and enumerate these cells on a large area on a microchip. The microfluidic chip and a lens-less imaging platform were then merged to monitor cell populations and morphologies as a system that may find use in distributed sensor networks. The chip, featuring surface chemistry and automatic cell imaging, was fabricated from a cover glass slide, double sided adhesive film and a transparent Polymethlymetacrylate (PMMA) slab. The optically clear chip allows detecting cells with a CCD sensor. These chips were fabricated with a laser cutter without the use of photolithography. We utilized CD4+ cells that are captured on the floor of a microfluidic chip due to the ability to address specific target cells using antibody-antigen binding. Captured CD4+ cells were imaged with a fluorescence microscope to verify the chip specificity and efficiency. We achieved 70.2 +/- 6.5% capturing efficiency and 88.8 +/- 5.4% specificity for CD4+ T lymphocytes (n = 9 devices). Bright field images of the captured cells in the 24 mm × 4 mm × 50 μm microfluidic chip were obtained with the CCD sensor in one second. We achieved an inexpensive system that rapidly captures cells and images them using a lens-less CCD system. This microfluidic device can be modified for use in single cell detection utilizing a cheap light-emitting diode (LED) chip instead of a wide range CCD system.

  17. Biomarker evaluation of face transplant rejection: association of donor T cells with target cell injury.

    PubMed

    Lian, Christine Guo; Bueno, Ericka M; Granter, Scott R; Laga, Alvaro C; Saavedra, Arturo P; Lin, William M; Susa, Joseph S; Zhan, Qian; Chandraker, Anil K; Tullius, Stefan G; Pomahac, Bohdan; Murphy, George F

    2014-06-01

    This series of 113 sequential biopsies of full facial transplants provides findings of potential translational significance as well as biological insights that could prompt reexamination of conventional paradigms of effector pathways in skin allograft rejection. Serial biopsies before, during, and after rejection episodes were evaluated for clinicopathological assessment that in selected cases included specific biomarkers for donor-versus-recipient T cells. Histologic evidence of rejection included lymphocyte-associated injury to epidermal rete ridges, follicular infundibula, and dermal microvessels. Surprisingly, during active rejection, immune cells spatially associated with target cell injury consisted abundantly or predominantly of lymphocytes of donor origin with an immunophenotype typical of the resident memory T-cell subset. Current dogma assumes that skin allograft rejection is mediated by recipient T cells that attack epidermal targets, and the association of donor T cells with sites of target cell injury raises questions regarding the potential complexity of immune cell interactions in the rejection process. A more histopathologically refined and immune-based biomarker approach to assessment of rejection of facial transplants is now indicated.

  18. Efficient Generation of Somatic Cell Nuclear Transfer-Competent Porcine Cells with Mutated Alleles at Multiple Target Loci by Using CRISPR/Cas9 Combined with Targeted Toxin-Based Selection System.

    PubMed

    Sato, Masahiro; Miyoshi, Kazuchika; Nakamura, Shingo; Ohtsuka, Masato; Sakurai, Takayuki; Watanabe, Satoshi; Kawaguchi, Hiroaki; Tanimoto, Akihide

    2017-12-04

    The recent advancement in genome editing such a CRISPR/Cas9 system has enabled isolation of cells with knocked multiple alleles through a one-step transfection. Somatic cell nuclear transfer (SCNT) has been frequently employed as one of the efficient tools for the production of genetically modified (GM) animals. To use GM cells as SCNT donor, efficient isolation of transfectants with mutations at multiple target loci is often required. The methods for the isolation of such GM cells largely rely on the use of drug selection-based approach using selectable genes; however, it is often difficult to isolate cells with mutations at multiple target loci. In this study, we used a novel approach for the efficient isolation of porcine cells with at least two target loci mutations by one-step introduction of CRISPR/Cas9-related components. A single guide (sg) RNA targeted to GGTA1 gene, involved in the synthesis of cell-surface α-Gal epitope (known as xenogenic antigen), is always a prerequisite. When the transfected cells were reacted with toxin-labeled BS-I-B₄ isolectin for 2 h at 37 C to eliminate α-Gal epitope-expressing cells, the surviving clones lacked α-Gal epitope expression and were highly expected to exhibit induced mutations at another target loci. Analysis of these α-Gal epitope-negative surviving cells demonstrated a 100% occurrence of genome editing at target loci. SCNT using these cells as donors resulted in the production of cloned blastocysts with the genotype similar to that of the donor cells used. Thus, this novel system will be useful for SCNT-mediated acquisition of GM cloned piglets, in which multiple target loci may be mutated.

  19. Curcumin suppresses proliferation of colon cancer cells by targeting CDK2.

    PubMed

    Lim, Tae-Gyu; Lee, Sung-Young; Huang, Zunnan; Lim, Do Young; Chen, Hanyong; Jung, Sung Keun; Bode, Ann M; Lee, Ki Won; Dong, Zigang

    2014-04-01

    Curcumin, the yellow pigment of turmeric found in Southeast Indian food, is one of the most popular phytochemicals for cancer prevention. Numerous reports have demonstrated modulation of multiple cellular signaling pathways by curcumin and its molecular targets in various cancer cell lines. To identify a new molecular target of curcumin, we used shape screening and reverse docking to screen the Protein Data Bank against curcumin. Cyclin-dependent kinase 2 (CDK2), a major cell-cycle protein, was identified as a potential molecular target of curcumin. Indeed, in vitro and ex vivo kinase assay data revealed a dramatic suppressive effect of curcumin on CDK2 kinase activity. Furthermore, curcumin induced G1 cell-cycle arrest, which is regulated by CDK2 in HCT116 cells. Although the expression levels of CDK2 and its regulatory subunit, cyclin E, were not changed, the phosphorylation of retinoblastoma (Rb), a well-known CDK2 substrate, was reduced by curcumin. Because curcumin induced cell-cycle arrest, we investigated the antiproliferative effect of curcumin on HCT116 colon cancer cells. In this experiment, curcumin suppressed HCT116 cell proliferation effectively. To determine whether CDK2 is a direct target of curcumin, CDK2 expression was knocked down in HCT116 cells. As expected, HCT116 sh-CDK2 cells exhibited G1 arrest and reduced proliferation. Because of the low levels of CDK2 in HCT116 sh-CDK2 cells, the effects of curcumin on G1 arrest and cell proliferation were not substantially relative to HCT116 sh-control cells. From these results, we identified CDK2 as a direct target of curcumin in colon cancer cells.

  20. Curcumin suppresses proliferation of colon cancer cells by targeting CDK2

    PubMed Central

    Lim, Tae-Gyu; Lee, Sung-Young; Huang, Zunnan; Lim, Do Young; Chen, Hanyong; Jung, Sung Keun; Bode, Ann M.; Lee, Ki Won; Dong, Zigang

    2014-01-01

    Curcumin, the yellow pigment of turmeric found in Southeast Indian food, is one of the most popular phytochemicals for cancer prevention. Numerous reports have demonstrated modulation of multiple cellular signaling pathways by curcumin and its molecular targets in various cancer cell lines. To identify a new molecular target of curcumin, we used shape screening and reverse docking to screen the protein data bank against curcumin. Cyclin dependent kinase 2 (CDK2), a major cell cycle protein, was identified as a potential molecular target of curcumin. Indeed, in vitro and ex vivo kinase assay data revealed a dramatic suppressive effect of curcumin on CDK2 kinase activity. Furthermore, curcumin induced G1 cell cycle arrest, which is regulated by CDK2 in HCT116 cells. Although the expression levels of CDK2 and its regulatory subunit, cyclin E, were not changed, the phosphorylation of Rb, a well-known CDK2 substrate, was reduced by curcumin. Because curcumin induced cell cycle arrest, we investigated the anti-proliferative effect of curcumin on HCT116 colon cancer cells. In this experiment, curcumin suppressed HCT116 cell proliferation effectively. To determine if CDK2 is a direct target of curcumin, CDK2 expression was knocked down in HCT116 cells. As expected, HCT116 sh-CDK2 cells exhibited G1 arrest and reduced proliferation. Because of the low levels of CDK2 in HCT116 sh-CDK2 cells, the effects of curcumin on G1 arrest and cell proliferation were not substantial relative to HCT116 sh-control cells. From these results, we identified CDK2 as a direct target of curcumin in colon cancer cells. PMID:24550143

  1. Disassembly of actin structures by nanosecond pulsed electric field is a downstream effect of cell swelling.

    PubMed

    Pakhomov, Andrei G; Xiao, Shu; Pakhomova, Olga N; Semenov, Iurii; Kuipers, Marjorie A; Ibey, Bennett L

    2014-12-01

    Disruption of the actin cytoskeleton structures was reported as one of the characteristic effects of nanosecond-duration pulsed electric field (nsPEF) in both mammalian and plant cells. We utilized CHO cells that expressed the monomeric fluorescent protein (mApple) tagged to actin to test if nsPEF modifies the cell actin directly or as a consequence of cell membrane permeabilization. A train of four 600-ns pulses at 19.2 kV/cm (2 Hz) caused immediate cell membrane poration manifested by YO-PRO-1 dye uptake, gradual cell rounding and swelling. Concurrently, bright actin features were replaced by dimmer and uniform fluorescence of diffuse actin. To block the nsPEF-induced swelling, the bath buffer was isoosmotically supplemented with an electropore-impermeable solute (sucrose). A similar addition of a smaller, electropore-permeable solute (adonitol) served as a control. We demonstrated that sucrose efficiently blocked disassembly of actin features by nsPEF, whereas adonitol did not. Sucrose also attenuated bleaching of mApple-tagged actin in nsPEF-treated cells (as integrated over the cell volume), although did not fully prevent it. We conclude that disintegration of the actin cytoskeleton was a result of cell swelling, which, in turn, was caused by cell permeabilization by nsPEF and transmembrane diffusion of solutes which led to the osmotic imbalance. Copyright © 2014 Elsevier B.V. All rights reserved.

  2. Enhancing Oral Vaccine Potency by Targeting Intestinal M Cells

    PubMed Central

    Azizi, Ali; Kumar, Ashok; Diaz-Mitoma, Francisco; Mestecky, Jiri

    2010-01-01

    The immune system in the gastrointestinal tract plays a crucial role in the control of infection, as it constitutes the first line of defense against mucosal pathogens. The attractive features of oral immunization have led to the exploration of a variety of oral delivery systems. However, none of these oral delivery systems have been applied to existing commercial vaccines. To overcome this, a new generation of oral vaccine delivery systems that target antigens to gut-associated lymphoid tissue is required. One promising approach is to exploit the potential of microfold (M) cells by mimicking the entry of pathogens into these cells. Targeting specific receptors on the apical surface of M cells might enhance the entry of antigens, initiating the immune response and consequently leading to protection against mucosal pathogens. In this article, we briefly review the challenges associated with current oral vaccine delivery systems and discuss strategies that might potentially target mouse and human intestinal M cells. PMID:21085599

  3. Microcinematographic and electron microscopic analysis of target cell lysis induced by cytotoxic T lymphocytes.

    PubMed Central

    Matter, A

    1979-01-01

    A study was carried out to determine the sequence of events of T-cell mediated target cell lysis in microcinematography and electron microscopy. Highly efficient cytotoxic T lymphocytes (CTL) were generated in vivo and in vitro using preimmunized spleen cells and purification procedures. Such CTL were highly specific. This specificity correlated well with the number of adhesions formed between CTL and targets and this criterion was used to study killer-target cell interaction. Microcinematography showed that target cell lysis at the single cell level, despite time variations, could be clearly separated into three phases: (a) a recognition phase, visible by random crawling of CTL over the target cell surface until firm contact was established; (b) a post-recognition phase, during which firm contact between CTL and target was maintained without gross modification of either cell; (c) a phase of target cell disintegration, mainly characterized by vigorous blebbing of the cell membrane resulting in a motionless carcass of the target cell but not in its total dissolution. Only later this carcass decayed and formed a necrotic ghost. Electron microscopic observations were put into sequence according to microcinematography. Post-recognition phase was characterized by a tight apposition of the membranes of CTL and target cell. No gap junctions could be observed. During target cell disintegration, profound cytoplasmic and nuclear changes occurred simultaneous with surface blebbing. Most noticeable were extensive internal vacuolization, mitochondrial swelling, nuclear pycnosis and dissolution of the nucleolus. These observations suggested that target cell lysis does not start with a surface phenomenon similar to complement lysis, but a process involving practically the whole cell simultaneously. It is conceivable, therefore, that the signal from the CTL is transmitted across the target cell, and that the switch to sudden cell death is manipulated deep inside the cell. Images

  4. Colon-targeted delivery of live bacterial cell biotherapeutics including microencapsulated live bacterial cells

    PubMed Central

    Prakash, Satya; Malgorzata Urbanska, Aleksandra

    2008-01-01

    There has been an ample interest in delivery of therapeutic molecules using live cells. Oral delivery has been stipulated as best way to deliver live cells to humans for therapy. Colon, in particular, is a part of gastrointestinal (GI) tract that has been proposed to be an oral targeted site. The main objective of these oral therapy procedures is to deliver live cells not only to treat diseases like colorectal cancer, inflammatory bowel disease, and other GI tract diseases like intestinal obstruction and gastritis, but also to deliver therapeutic molecules for overall therapy in various diseases such as renal failure, coronary heart disease, hypertension, and others. This review provides a comprehensive summary of recent advancement in colon targeted live bacterial cell biotherapeutics. Current status of bacterial cell therapy, principles of artificial cells and its potentials in oral delivery of live bacterial cell biotherapeutics for clinical applications as well as biotherapeutic future perspectives are also discussed in our review. PMID:19707368

  5. The effect of graphitic target density on carbon nanotube synthesis by pulsed laser ablation method

    NASA Astrophysics Data System (ADS)

    Kazeimzadeh, Fatemeh; Malekfar, Rasoul; Houshiar, Mahboubeh

    2018-01-01

    Carbon nanotube (CNT) was synthesized by pulsed laser ablation (PLA) of a graphitic target in vacuum chamber filled by argon gas. The effect of different condition of target preparation on the amount and quality of carbon nanotube generation was investigated. The graphite powder with 2 at% micrometer nickel (Ni) powder was mixed and packed in to a mold using a hydraulic press device at a pressure of 1000 kg/cm3. The obtained pellet which contained the mixture powder provided the carbon source for CNTs formation in PLA method. Two pellets with the pressure time of 15 and 200 min was prepared. It has been shown that the time which graphitic target is under pressure is an effective parameter that can increase the amount of produced CNTs. Field emission scanning electron microscopy (FESEM) images show that if the density of graphitic target is increased by raising up the pressure time, CNTs can grow even under the condition in which usually no nanotube can be formed. It can be due to the elimination of the distances between the graphite and catalyst grains that causes the catalysis performance improvement. The experiment was performed for nanometer cobalt ferrite (CoFe2O4) together with Ni catalyst particles too. The diameter of synthesized CNPs was larger in the case of pure nickel that is related to the size of catalysts. Moreover, it was also observed that the production rate of the nanotubes increased for high density targets. This shows that the results are independent of the type of catalyst.

  6. Analysis of short pulse laser altimetry data obtained over horizontal path

    NASA Technical Reports Server (NTRS)

    Im, K. E.; Tsai, B. M.; Gardner, C. S.

    1983-01-01

    Recent pulsed measurements of atmospheric delay obtained by ranging to the more realistic targets including a simulated ocean target and an extended plate target are discussed. These measurements are used to estimate the expected timing accuracy of a correlation receiver system. The experimental work was conducted using a pulsed two color laser altimeter.

  7. Targeting Notch signalling pathway of cancer stem cells.

    PubMed

    Venkatesh, Vandana; Nataraj, Raghu; Thangaraj, Gopenath S; Karthikeyan, Murugesan; Gnanasekaran, Ashok; Kaginelli, Shanmukhappa B; Kuppanna, Gobianand; Kallappa, Chandrashekrappa Gowdru; Basalingappa, Kanthesh M

    2018-01-01

    Cancer stem cells (CSCs) have been defined as cells within tumor that possess the capacity to self-renew and to cause the heterogeneous lineages of cancer cells that comprise the tumor. CSCs have been increasingly identified in blood cancer, prostate, ovarian, lung, melanoma, pancreatic, colon, brain and many more malignancies. CSCs have slow growth rate and are resistant to chemotherapy and radiotherapy that lead to the failure of traditional current therapy. Eradicating the CSCs and recurrence, is promising aspect for the cure of cancer. The CSCs like any other stem cells activate the signal transduction pathways that involve the development and tissue homeostasis, which include Notch signaling pathway. The new treatment targets these pathway that control stem-cell replication, survival and differentiation that are under development. Notch inhibitors either single or in combination with chemotherapy drugs have been developed to treat cancer and its recurrence. This approach of targeting signaling pathway of CSCs represents a promising future direction for the therapeutic strategy to cure cancer.

  8. Development of pulsed processes for the manufacture of solar cells

    NASA Technical Reports Server (NTRS)

    Minnucci, J. A.

    1979-01-01

    Low-energy ion implantation processes for the automated production of silicon solar cells were investigated. Phosphorus ions at an energy of 10 keV and dose of 2 x 10 to the 15th power/sq cm were implanted in silicon solar cells to produce junctions, while boron ions at 25 keV and 5 x 10 to the 15th power were implanted in the cells to produce effective back surface fields. An ion implantation facility with a beam current up to 4 mA and a production throughput of 300 wafers per hour was designed and installed. A design was prepared for a 100 mA, automated implanter with a production capacity of 100 MW sub e/sq cm per year. Two process sequences were developed which employ ion implantation and furnace or pulse annealing. A computer program was used to determine costs for junction formation by ion implantation and various furnace annealing cycles to demonstrate cost effectiveness of these methods.

  9. Laser activated nanothermolysis of leukemia cells monitored by photothermal microscopy

    NASA Astrophysics Data System (ADS)

    Lapotko, Dmitri; Lukianova, Ekaterina; Shnip, Alexander; Zheltov, George; Potapnev, Michail; Savitsky, Valeriy; Klimovich, Olga; Oraevsky, Alexander

    2005-04-01

    We are developing new diagnostic and therapeutic technologies for leukemia based on selective targeting of leukemia cells with gold nanoparticles and thermomechanical destruction of the tumor cells with laser-induced microbubbles. Clusters of spherical gold nanoparticles that have strong optical absorption of laser pulses at 532 nm served as nucleation sites of vapor microbubbles. The nanoparticles were targeted selectively to leukemia cells using leukemia-specific surface receptors and a set of two monoclonal antibodies. Application of a primary myeloid-specific antibody to tumor cells followed by targeting the cells with 30-nm nanoparticles conjugated with a secondary antibody (IgG) resulted in formation of nanoparticulate clusters due to aggregation of IgGs. Formation of clusters resulted in substantial decrease of the damage threshold for target cells. The results encourage development of Laser Activated Nanothermolysis as a Cell Elimination Therapy (LANCET) for leukemia. The proposed technology can be applied separately or in combination with chemotherapy for killing leukemia cells without damage to other blood cells. Potential applications include initial reduction of concentration of leukemia cells in blood prior to chemotherapy and treatment of residual tumor cells after the chemotherapy. Laser-induced bubbles in individual cells and cell damage were monitored by analyzing profile of photothermal response signals over the entire cell after irradiation with a single 10-ns long laser pulse. Photothermal microscopy was utilized for imaging formation of microbubbles around nanoparticulate clusters.

  10. Preparation of W–Sc{sub 2}O{sub 3} targets and scandate cathodes with film prepared by pulsed laser deposition

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    Zhang, Xizhu; Wang, Jinshu, E-mail: wangjsh@bjut.edu.cn; Liu, Wei

    2013-12-15

    Graphical abstract: - Highlights: • W–Sc{sub 2}O{sub 3} film containing 5% Sc{sub 2}O{sub 3} and 95% W were prepared by pulsed laser deposition. • W–Sc{sub 2}O{sub 3} film on scandate cathode surface improves emission property. • The film improves Sc distribution uniformity and is favorable for forming Ba–Sc–O layer. - Abstract: Sub-micrometer Sc{sub 2}O{sub 3}–W powder with a narrow particle size distribution has been obtained by a sol–gel method combined with two-step hydrogen reduction process. Based on the obtained powder, the W–Sc{sub 2}O{sub 3} targets have been sintered via spark plasma sintering (SPS) at 1300 °C. The W–Sc{sub 2}O{sub 3}more » targets have the average grain size of about 1 μm. Both the sintering temperature and holding time are much lower than those of the targets prepared with micrometer sized powders. The obtained W–Sc{sub 2}O{sub 3} targets have a high comparative density of 96.4% and rockwell hardness of 86.4 HRC. Using the target, the scandate cathode deposited with a film containing 5% Sc{sub 2}O{sub 3} and 95% W has been obtained by pulsed laser deposition (PLD) method. This cathode has good emission property, i.e., the highest thermionic emission current density reaches 43.09 A/cm{sup 2} of J{sub div} at 900 °C{sub b} after being activated for 8 h, which is much higher than that of scandate cathode without film. Scandium (Sc) supplied by the film on the surface during the activation forms a Ba–Sc–O active layer, which helps to the emission.« less

  11. Ultra-intense laser interaction with specially-designed targets as a source of energetic protons

    NASA Astrophysics Data System (ADS)

    Psikal, J.; Matys, M.

    2017-05-01

    In this contribution, we discuss the optimization of laser driven proton acceleration efficiency by nanostructured targets, interpret the experimental results showing the manipulation of proton beam profiles by nanosctructured rear surface of the targets and investigate the acceleration of protons from hydrogen solid ribbon by PW-class lasers, with the help of multidimensional particle-in-cell simulations. Microstructured hollow targets are proposed to enhance the absorption of the laser pulse energy while keeping the target thickness to minimum, which is both favorable for enhanced efficiency of the acceleration of protons. Thin targets with grating structures of various configurations on their rear sides stretch the proton beams in the perpendicular direction to the grating orientation due to transverse electric fields generated inside the target grooves and can reduce the proton beam divergence in the parallel direction to the grating due to a lower density of the stretched beam compared with flat foils. Finally, it is shown that when multiPW laser pulse interacts with hydrogen solid ribbon, hole boring radiation pressure acceleration (RPA) dominates over the target normal sheath acceleration (TNSA).

  12. Pulsed power molten salt battery

    NASA Technical Reports Server (NTRS)

    Argade, Shyam D.

    1992-01-01

    It was concluded that carbon cathodes with chlorine work well. Lithium alloy chlorine at 450 C, 1 atm given high power capability, high energy density, DC + pulsing yields 600 pulses, no initial peak, and can go to red heat without burn-up. Electrochemical performance at the cell and cell stack level out under demanding test regime. Engineering and full prototype development for advancing this technology is warranted.

  13. A new prospect in cancer therapy: targeting cancer stem cells to eradicate cancer.

    PubMed

    Chen, Li-Sha; Wang, An-Xin; Dong, Bing; Pu, Ke-Feng; Yuan, Li-Hua; Zhu, Yi-Min

    2012-12-01

    According to the cancer stem cell theory, cancers can be initiated by cancer stem cells. This makes cancer stem cells prime targets for therapeutic intervention. Eradicating cancer stem cells by efficient targeting agents may have the potential to cure cancer. In this review, we summarize recent breakthroughs that have improved our understanding of cancer stem cells, and we discuss the therapeutic strategy of targeting cancer stem cells, a promising future direction for cancer stem cell research.

  14. Glioblastoma Stem Cells as a New Therapeutic Target for Glioblastoma.

    PubMed

    Kalkan, Rasime

    2015-01-01

    Primary and secondary glioblastomas (GBMs) are two distinct diseases. The genetic and epigenetic background of these tumors is highly variable. The treatment procedure for these tumors is often unsuccessful because of the cellular heterogeneity and intrinsic ability of the tumor cells to invade healthy tissues. The fatal outcome of these tumors promotes researchers to find out new markers associated with the prognosis and treatment planning. In this communication, the role of glioblastoma stem cells in tumor progression and the malignant behavior of GBMs are summarized with attention to the signaling pathways and molecular regulators that are involved in maintaining the glioblastoma stem cell phenotype. A better understanding of these stem cell-like cells is necessary for designing new effective treatments and developing novel molecular strategies to target glioblastoma stem cells. We discuss hypoxia as a new therapeutic target for GBM. We focus on the inhibition of signaling pathways, which are associated with the hypoxia-mediated maintenance of glioblastoma stem cells, and the knockdown of hypoxia-inducible factors, which could be identified as attractive molecular target approaches for GBM therapeutics.

  15. Monoclonal antibodies targeting non-small cell lung cancer stem-like cells by multipotent cancer stem cell monoclonal antibody library.

    PubMed

    Cao, Kaiyue; Pan, Yunzhi; Yu, Long; Shu, Xiong; Yang, Jing; Sun, Linxin; Sun, Lichao; Yang, Zhihua; Ran, Yuliang

    2017-02-01

    Cancer stem cells (CSCs) are a rare subset of cancer cells that play a significant role in cancer initiation, spreading, and recurrence. In this study, a subpopulation of lung cancer stem-like cells (LCSLCs) was identified from non-small cell lung carcinoma cell lines, SPCA-1 and A549, using serum-free suspension sphere-forming culture method. A monoclonal antibody library was constructed using immunized BLAB/c mice with the multipotent CSC cell line T3A-A3. Flow cytometry analysis showed that 33 mAbs targeted antigens can be enriched in sphere cells compared with the parental cells of SPCA-1 and A549 cell lines. Then, we performed functional antibody screening including sphere-forming inhibiting and invasion inhibiting assay. The results showed that two antibodies, 12C7 and 9B8, notably suppressed the self-renewal and invasion of LCSLCs. Fluorescence-activated cell sorting (FACs) found that the positive cells recognized by mAbs, 12C7 or 9B8, displayed features of LCSLCs. Interestingly, we found that these two antibodies recognized different subsets of cells and their combination effect was superior to the individual effect both in vitro and in vivo. Tissue microarrays were applied to detect the expression of the antigens targeted by these two antibodies. The positive expression of 12C7 and 9B8 targeted antigen was 84.4 and 82.5%, respectively, which was significantly higher than that in the non-tumor lung tissues. In conclusion, we screened two potential therapeutic antibodies that target different subsets of LCSLCs.

  16. A new pulsed laser deposition technique: scanning multi-component pulsed laser deposition method.

    PubMed

    Fischer, D; de la Fuente, G F; Jansen, M

    2012-04-01

    The scanning multi-component pulsed laser deposition (PLD) method realizes uniform depositions of desired coatings by a modified pulsed laser deposition process, preferably with a femto-second laser-system. Multi-component coatings (single or multilayered) are thus deposited onto substrates via laser induced ablation of segmented targets. This is achieved via horizontal line-scanning of a focused laser beam over a uniformly moving target's surface. This process allows to deposit the desired composition of the coating simultaneously, starting from the different segments of the target and adjusting the scan line as a function of target geometry. The sequence and thickness of multilayers can easily be adjusted by target architecture and motion, enabling inter/intra layer concentration gradients and thus functional gradient coatings. This new, simple PLD method enables the achievement of uniform, large-area coatings. Case studies were performed with segmented targets containing aluminum, titanium, and niobium. Under the laser irradiation conditions applied, all three metals were uniformly ablated. The elemental composition within the rough coatings obtained was fixed by the scanned area to Ti-Al-Nb = 1:1:1. Crystalline aluminum, titanium, and niobium were found to coexist side by side at room temperature within the substrate, without alloy formation up to 600 °C. © 2012 American Institute of Physics

  17. HLA-targeted flow cytometric sorting of blood cells allows separation of pure and viable microchimeric cell populations.

    PubMed

    Drabbels, Jos J M; van de Keur, Carin; Kemps, Berit M; Mulder, Arend; Scherjon, Sicco A; Claas, Frans H J; Eikmans, Michael

    2011-11-10

    Microchimerism is defined by the presence of low levels of nonhost cells in a person. We developed a reliable method for separating viable microchimeric cells from the host environment. For flow cytometric cell sorting, HLA antigens were targeted with human monoclonal HLA antibodies (mAbs). Optimal separation of microchimeric cells (present at a proportion as low as 0.01% in artificial mixtures) was obtained with 2 different HLA mAbs, one targeting the chimeric cells and the other the background cells. To verify purity of separated cell populations, flow-sorted fractions of 1000 cells were processed for DNA analysis by HLA-allele-specific and Y-chromosome-directed real-time quantitative PCR assays. After sorting, PCR signals of chimeric DNA markers in the positive fractions were significantly enhanced compared with those in the presort samples, and they were similar to those in 100% chimeric control samples. Next, we demonstrate applicability of HLA-targeted FACS sorting after pregnancy by separating chimeric maternal cells from child umbilical cord mononuclear cells. Targeting allelic differences with anti-HLA mAbs with FACS sorting allows maximal enrichment of viable microchimeric cells from a background cell population. The current methodology enables reliable microchimeric cell detection and separation in clinical specimens.

  18. Ultrashort pulse high intensity laser illumination of a simple metal

    NASA Astrophysics Data System (ADS)

    Milchberg, H. M.; Freeman, R. R.; Davey, S. C.

    1988-10-01

    We have observed the self-reflection of intense, sub-picosecond 308 nm light pulse incident on a planar Al target and have inferred the electrical conductivity of solid density Al. The pulse lengths were sufficiently short that no significant expansion of the target occurred during the measurement.

  19. Zn(II)-curc targets p53 in thyroid cancer cells.

    PubMed

    Garufi, Alessia; D'Orazi, Valerio; Crispini, Alessandra; D'Orazi, Gabriella

    2015-10-01

    TP53 mutation is a common event in many cancers, including thyroid carcinoma. Defective p53 activity promotes cancer resistance to therapies and a more malignant phenotype, acquiring oncogenic functions. Rescuing the function of mutant p53 (mutp53) protein is an attractive anticancer therapeutic strategy. Zn(II)-curc is a novel small molecule that has been shown to target mutp53 protein in several cancer cells, but its effect in thyroid cancer cells remains unclear. Here, we investigated whether Zn(II)-curc could affect p53 in thyroid cancer cells with both p53 mutation (R273H) and wild-type p53. Zn(II)-curc induced mutp53H273 downregulation and reactivation of wild-type functions, such as binding to canonical target promoters and target gene transactivation. This latter effect was similar to that induced by PRIMA-1. In addition, Zn(II)-curc triggered p53 target gene expression in wild-type p53-carrying cells. In combination treatments, Zn(II)-curc enhanced the antitumor activity of chemotherapeutic drugs, in both mutant and wild-type-carrying cancer cells. Taken together, our data indicate that Zn(II)-curc promotes the reactivation of p53 in thyroid cancer cells, providing in vitro evidence for a potential therapeutic approach in thyroid cancers.

  20. ErbB-targeted CAR T-cell immunotherapy of cancer.

    PubMed

    Whilding, Lynsey M; Maher, John

    2015-01-01

    Chimeric antigen receptor (CAR) based immunotherapy has been under development for the last 25 years and is now a promising new treatment modality in the field of cancer immunotherapy. The approach involves genetically engineering T cells to target malignant cells through expression of a bespoke fusion receptor that couples an HLA-independent antigen recognition domain to one or more intracellular T-cell activating modules. Multiple clinical trials are now underway in several centers to investigate CAR T-cell immunotherapy of diverse hematologic and solid tumor types. The most successful results have been achieved in the treatment of patients with B-cell malignancies, in whom several complete and durable responses have been achieved. This review focuses on the preclinical and clinical development of CAR T-cell immunotherapy of solid cancers, targeted against members of the ErbB family.

  1. Comparison of pulse wave velocity and pulse pressure amplification in association with target organ damage in community-dwelling elderly: The Northern Shanghai Study.

    PubMed

    Bai, Bin; Teliewubai, Jiadela; Lu, Yuyan; Yu, Shikai; Xiong, Jing; Chi, Chen; Zhou, Yiwu; Ji, Hongwei; Fan, Ximin; Blacher, Jacques; Li, Jue; Zhang, Yi; Xu, Yawei

    2018-05-01

    This study aimed to investigate the discrepancy between pulse wave velocity (PWV) and pulse pressure amplification (PPA) in association with hypertensive target organ damage (TOD) in the elderly. From June 2014 to August 2015, 1599 participants aged >65 years old from communities located in northern Shanghai were recruited. Carotid-femoral pulse wave velocity (cfPWV), peripheral blood pressure (BP), central BP and other TOD indicators, including the ratio of the early ventricular filling velocity (E) to the peak velocity of the tissue Doppler velocity of septal mitral annulus (E/Ea), left ventricular mass index (LVMI), carotid intima-medium thickness (CIMT), estimated glomerular filtration rate (eGFR), and urinary albumin-creatinine ratio (ACR), were determined for each participant. PPA was defined as the peripheral-to-central pulse pressure ratio. In multivariable linear regression analysis, cfPWV was significantly associated with CIMT (β = 12.83 ± 4.28 μm per SD; P = 0.003) and eGFR (β = -1.85 ± 0.69 ml/min/1.73 m 2 per SD; P = 0.007), whereas PPA was significantly associated with E/Ea (β = -0.25 ± 0.10 per SD; P = 0.01) and LVMI (β = -3.00 ± 0.78 g/m 2 per SD; P < 0.001). Similarly, in multivariable logistic regression analysis, cfPWV was significantly associated with arterial plaque (odds ratio [OR], 1.21 [95% confidence interval [CI], 1.05-1.39]; P = 0.007), peripheral artery disease (OR, 1.22 [95% CI, 1.06-1.42]; P = 0.007), chronic kidney diseases (OR, 1.24 [95% CI, 1.01-1.54]; P = 0.04) and microalbuminuria (OR, 1.21 [95% CI, 1.07-1.37]; P = 0.002), while PPA was tightly associated with left ventricular hypertrophy (OR, 0.85 [95% CI, 0.72-0.99]; P = 0.04) and diastolic dysfunction (OR, 0.78 [95% CI, 0.64-0.96]; P = 0.02). In conclusion, cfPWV is a vessel-related and renal-related biomarker, while PPA is a cardiac-related biomarker in community-based elderly.

  2. Pulse stretcher

    DOEpatents

    Horton, James A.

    1994-01-01

    Apparatus (20) for increasing the length of a laser pulse to reduce its peak power without substantial loss in the average power of the pulse. The apparatus (20) uses a White cell (10) having a plurality of optical delay paths (18a-18d) of successively increasing number of passes between the field mirror (13) and the objective mirrors (11 and 12). A pulse (26) from a laser (27) travels through a multi-leg reflective path (28) between a beam splitter (21) and a totally reflective mirror (24) to the laser output (37). The laser pulse (26) is also simultaneously injected through the beam splitter (21) to the input mirrors (14a-14d) of the optical delay paths (18a-18d). The pulses from the output mirrors (16a-16d) of the optical delay paths (18a-18d) go simultaneously to the laser output (37) and to the input mirrors ( 14b-14d) of the longer optical delay paths. The beam splitter (21) is 50% reflective and 50% transmissive to provide equal attenuation of all of the pulses at the laser output (37).

  3. Timing of target discrimination in human frontal eye fields.

    PubMed

    O'Shea, Jacinta; Muggleton, Neil G; Cowey, Alan; Walsh, Vincent

    2004-01-01

    Frontal eye field (FEF) neurons discharge in response to behaviorally relevant stimuli that are potential targets for saccades. Distinct visual and motor processes have been dissociated in the FEF of macaque monkeys, but little is known about the visual processing capacity of FEF in humans. We used double-pulse transcranial magnetic stimulation [(d)TMS] to investigate the timing of target discrimination during visual conjunction search. We applied dual TMS pulses separated by 40 msec over the right FEF and vertex. These were applied in five timing conditions to sample separate time windows within the first 200 msec of visual processing. (d)TMS impaired search performance, reflected in reduced d' scores. This effect was limited to a time window between 40 and 80 msec after search array onset. These parameters correspond with single-cell activity in FEF that predicts monkeys' behavioral reports on hit, miss, false alarm, and correct rejection trials. Our findings demonstrate a crucial early role for human FEF in visual target discrimination that is independent of saccade programming.

  4. GEM-loaded magnetic albumin nanospheres modified with cetuximab for simultaneous targeting, magnetic resonance imaging, and double-targeted thermochemotherapy of pancreatic cancer cells.

    PubMed

    Wang, Ling; An, Yanli; Yuan, Chenyan; Zhang, Hao; Liang, Chen; Ding, Fengan; Gao, Qi; Zhang, Dongsheng

    2015-01-01

    Targeted delivery is a promising strategy to improve the diagnostic imaging and therapeutic effect of cancers. In this paper, novel cetuximab (C225)-conjugated, gemcitabine (GEM)-containing magnetic albumin nanospheres (C225-GEM/MANs) were fabricated and applied as a theranostic nanocarrier to conduct simultaneous targeting, magnetic resonance imaging (MRI), and double-targeted thermochemotherapy against pancreatic cancer cells. Fe3O4 nanoparticles (NPs) and GEM co-loaded albumin nanospheres (GEM/MANs) were prepared, and then C225 was further conjugated to synthesize C225-GEM/MANs. Their morphology, mean particle size, GEM encapsulation ratio, specific cell-binding ability, and thermal dynamic profiles were characterized. The effects of discriminating different EGFR-expressing pancreatic cancer cells (AsPC-1 and MIA PaCa-2) and monitoring cellular targeting effects were assessed by targeted MRI. Lastly, the antitumor efficiency of double/C225/magnetic-targeted and nontargeted thermochemotherapy was compared with chemotherapy alone using 3-(4, 5-dimethyl-2-thiazolyl)-2,5-diphenyl-2H-tetrazolium bromide (MTT) and flow cytometry (FCM) assay. When treated with targeted nanospheres, AsPC-1 cells showed a significantly less intense MRI T2 signal than MIA PaCa-2 cells, while both cells had similar signal strength when incubated with nontargeted nanospheres. T2 signal intensity was significantly lower when magnetic and C225 targeting were combined, rather than used alone. The inhibitory and apoptotic rates of each thermochemotherapy group were significantly higher than those of the chemotherapy-alone groups. Additionally, both MTT and FCM analysis verified that double-targeted thermochemotherapy had the highest targeted killing efficiency among all groups. The C225-GEM/MANs can distinguish various EGFR-expressing live pancreatic cancer cells, monitor diverse cellular targeting effects using targeted MRI imaging, and efficiently mediate double-targeted thermochemotherapy

  5. 3D pulsed laser-triggered high-speed microfluidic fluorescence-activated cell sorter

    PubMed Central

    Chen, Yue; Wu, Ting-Hsiang; Kung, Yu-Chun; Teitell, Michael A.; Chiou, Pei-Yu

    2014-01-01

    We report a 3D microfluidic pulsed laser-triggered fluorescence-activated cell sorter capable of sorting at a throughput of 23,000 cells sec−1 with 90% purity in high-purity mode and at a throughput of 45,000 cells sec−1 with 45% purity in enrichment mode in one stage and in a single channel. This performance is realized by exciting laser-induced cavitation bubbles in a 3D PDMS microfluidic channel to generate high-speed liquid jets that deflect detected fluorescent cells and particles focused by 3D sheath flows. The ultrafast switching mechanism (20 μsec complete on-off cycle), small liquid jet perturbation volume, and three-dimensional sheath flow focusing for accurate timing control of fast (1.5 m sec−1) passing cells and particles are three critical factors enabling high-purity sorting at high-throughput in this sorter. PMID:23844418

  6. Development of the dense plasma focus for short-pulse applications

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    Bennett, N.; Blasco, M.; Breeding, K.

    The dense plasma focus (DPF) has long been considered a compact source for pulsed neutrons and has traditionally been optimized for the total neutron yield. Here, we describe the efforts to optimize the DPF for short-pulse applications by introducing a reentrant cathode at the end of the coaxial plasma gun. We reduced the resulting neutron pulse widths by an average of 21±921±9% from the traditional long-drift DPF design. Pulse widths and yields achieved from deuterium-tritium fusion at 2 MA are 61.8±30.761.8±30.7 ns FWHM and 1.84±0.49×10121.84±0.49×10 12 neutrons per shot. Simulations were conducted concurrently to elucidate the DPF operation and confirmmore » the role of the reentrant cathode. Furthermore, a hybrid fluid-kinetic particle-in-cell modeling capability demonstrates correct sheath velocities, plasma instabilities, and fusion yield rates. Consistent with previous findings that the DPF is dominated by beam-target fusion from superthermal ions, we estimate that the thermonuclear contribution is at the 1% level.« less

  7. Development of the dense plasma focus for short-pulse applications

    DOE PAGES

    Bennett, N.; Blasco, M.; Breeding, K.; ...

    2017-01-05

    The dense plasma focus (DPF) has long been considered a compact source for pulsed neutrons and has traditionally been optimized for the total neutron yield. Here, we describe the efforts to optimize the DPF for short-pulse applications by introducing a reentrant cathode at the end of the coaxial plasma gun. We reduced the resulting neutron pulse widths by an average of 21±921±9% from the traditional long-drift DPF design. Pulse widths and yields achieved from deuterium-tritium fusion at 2 MA are 61.8±30.761.8±30.7 ns FWHM and 1.84±0.49×10121.84±0.49×10 12 neutrons per shot. Simulations were conducted concurrently to elucidate the DPF operation and confirmmore » the role of the reentrant cathode. Furthermore, a hybrid fluid-kinetic particle-in-cell modeling capability demonstrates correct sheath velocities, plasma instabilities, and fusion yield rates. Consistent with previous findings that the DPF is dominated by beam-target fusion from superthermal ions, we estimate that the thermonuclear contribution is at the 1% level.« less

  8. Propulsion of a flat tin target with pulsed CO2 laser radiation: measurements using a ballistic pendulum

    NASA Astrophysics Data System (ADS)

    Lakatosh, B. V.; Abramenko, D. B.; Ivanov, V. V.; Medvedev, V. V.; Krivtsun, V. M.; Koshelev, K. N.; Yakunin, A. M.

    2018-01-01

    The recoil momentum transfer produced by irradiation of a flat tin (Sn) target with pulses of high-power CO2 laser with intensity ranging from 107 to 1010 W cm-2 has been studied. Momentum measurements were performed using a ballistic pendulum, capable of measuring momenta as small as 0.001 g · cm s-1 . It has been established that the recoil momentum monotonically increases with the laser energy and asymptotically reaches the power scaling law p ∼ Iα with α = 0.96 +/- 0.07 . Results are compared with previously published theoretical studies.

  9. Phenotypic high-throughput screening elucidates target pathway in breast cancer stem cell-like cells.

    PubMed

    Carmody, Leigh C; Germain, Andrew R; VerPlank, Lynn; Nag, Partha P; Muñoz, Benito; Perez, Jose R; Palmer, Michelle A J

    2012-10-01

    Cancer stem cells (CSCs) are resistant to standard cancer treatments and are likely responsible for cancer recurrence, but few therapies target this subpopulation. Due to the difficulty in propagating CSCs outside of the tumor environment, previous work identified CSC-like cells by inducing human breast epithelial cells into an epithelial-to-mesenchymal transdifferentiated state (HMLE_sh_ECad). A phenotypic screen was conducted against HMLE_sh_ECad with 300 718 compounds from the Molecular Libraries Small Molecule Repository to identify selective inhibitors of CSC growth. The screen yielded 2244 hits that were evaluated for toxicity and selectivity toward an isogenic control cell line. An acyl hydrazone scaffold emerged as a potent and selective scaffold targeting HMLE_sh_ECad. Fifty-three analogues were acquired and tested; compounds ranged in potency from 790 nM to inactive against HMLE_sh_ECad. Of the analogues, ML239 was best-in-class with an IC(50)= 1.18 µM against HMLE_sh_ECad, demonstrated a >23-fold selectivity over the control line, and was toxic to another CSC-like line, HMLE_shTwist, and a breast carcinoma cell line, MDA-MB-231. Gene expression studies conducted with ML239-treated cells showed altered gene expression in the NF-κB pathway in the HMLE_sh_ECad line but not in the isogenic control line. Future studies will be directed toward the identification of ML239 target(s).

  10. Wavelength Locking to CO2 Absorption Line-Center for 2-Micron Pulsed IPDA Lidar Application

    NASA Technical Reports Server (NTRS)

    Refaat, Tamer F.; Petros, Mulugeta; Antill, Charles W.; Singh, Upendra N.; Yu, Jirong

    2016-01-01

    An airborne 2-micron triple-pulse integrated path differential absorption (IPDA) lidar is currently under development at NASA Langley Research Center (LaRC). This IPDA lidar system targets both atmospheric carbon dioxide (CO2) and water vapor (H2O) column measurements. Independent wavelength control of each of the transmitted laser pulses is a key feature for the success of this instrument. The wavelength control unit provides switching, tuning and locking for each pulse in reference to a 2-micron CW (Continuous Wave) laser source locked to CO2 line-center. Targeting the CO2 R30 line center, at 2050.967 nanometers, a wavelength locking unit has been integrated using semiconductor laser diode. The CO2 center-line locking unit includes a laser diode current driver, temperature controller, center-line locking controller and CO2 absorption cell. This paper presents the CO2 center-line locking unit architecture, characterization procedure and results. Assessment of wavelength jitter on the IPDA measurement error will also be addressed by comparison to the system design.

  11. Detecting drug-target binding in cells using fluorescence-activated cell sorting coupled with mass spectrometry analysis.

    PubMed

    Wilson, Kris; Webster, Scott P; Iredale, John P; Zheng, Xiaozhong; Homer, Natalie Z; Pham, Nhan T; Auer, Manfred; Mole, Damian J

    2017-12-15

    The assessment of drug-target engagement for determining the efficacy of a compound inside cells remains challenging, particularly for difficult target proteins. Existing techniques are more suited to soluble protein targets. Difficult target proteins include those with challenging in vitro solubility, stability or purification properties that preclude target isolation. Here, we report a novel technique that measures intracellular compound-target complex formation, as well as cellular permeability, specificity and cytotoxicity-the toxicity-affinity-permeability-selectivity (TAPS) technique. The TAPS assay is exemplified here using human kynurenine 3-monooxygenase (KMO), a challenging intracellular membrane protein target of significant current interest. TAPS confirmed target binding of known KMO inhibitors inside cells. We conclude that the TAPS assay can be used to facilitate intracellular hit validation on most, if not all intracellular drug targets.

  12. Detecting drug-target binding in cells using fluorescence-activated cell sorting coupled with mass spectrometry analysis

    NASA Astrophysics Data System (ADS)

    Wilson, Kris; Webster, Scott P.; Iredale, John P.; Zheng, Xiaozhong; Homer, Natalie Z.; Pham, Nhan T.; Auer, Manfred; Mole, Damian J.

    2018-01-01

    The assessment of drug-target engagement for determining the efficacy of a compound inside cells remains challenging, particularly for difficult target proteins. Existing techniques are more suited to soluble protein targets. Difficult target proteins include those with challenging in vitro solubility, stability or purification properties that preclude target isolation. Here, we report a novel technique that measures intracellular compound-target complex formation, as well as cellular permeability, specificity and cytotoxicity-the toxicity-affinity-permeability-selectivity (TAPS) technique. The TAPS assay is exemplified here using human kynurenine 3-monooxygenase (KMO), a challenging intracellular membrane protein target of significant current interest. TAPS confirmed target binding of known KMO inhibitors inside cells. We conclude that the TAPS assay can be used to facilitate intracellular hit validation on most, if not all intracellular drug targets.

  13. Targeting myeloid-derived suppressor cells for cancer immunotherapy.

    PubMed

    Liu, Yijun; Wei, Guowei; Cheng, Wesley A; Dong, Zhenyuan; Sun, Han; Lee, Vincent Y; Cha, Soung-Chul; Smith, D Lynne; Kwak, Larry W; Qin, Hong

    2018-05-31

    Myeloid-derived suppressor cells (MDSCs) are a heterogeneous population of immature myeloid cells with an immune suppressive phenotype. They represent a critical component of the immune suppressive niche described in cancer, where they support immune escape and tumor progression through direct effects on both the innate and adaptive immune responses, largely by contributing to maintenance of a high oxidative stress environment. The number of MDSCs positively correlates with protumoral activity, and often diminishes the effectiveness of immunotherapies, which is particularly problematic with the emergence of personalized medicine. Approaches targeting MDSCs showed promising results in preclinical studies and are under active investigation in clinical trials in combination with various immune checkpoint inhibitors. In this review, we discuss MDSC targets and therapeutic approaches targeting MDSC that have the aim of enhancing the existing tumor therapies.

  14. Many si/shRNAs can kill cancer cells by targeting multiple survival genes through an off-target mechanism

    PubMed Central

    van Dongen, Stijn; Haluck-Kangas, Ashley; Sarshad, Aishe A; Bartom, Elizabeth T; Kim, Kwang-Youn A; Scholtens, Denise M; Hafner, Markus; Zhao, Jonathan C; Murmann, Andrea E

    2017-01-01

    Over 80% of multiple-tested siRNAs and shRNAs targeting CD95 or CD95 ligand (CD95L) induce a form of cell death characterized by simultaneous activation of multiple cell death pathways preferentially killing transformed and cancer stem cells. We now show these si/shRNAs kill cancer cells through canonical RNAi by targeting the 3’UTR of critical survival genes in a unique form of off-target effect we call DISE (death induced by survival gene elimination). Drosha and Dicer-deficient cells, devoid of most miRNAs, are hypersensitive to DISE, suggesting cellular miRNAs protect cells from this form of cell death. By testing 4666 shRNAs derived from the CD95 and CD95L mRNA sequences and an unrelated control gene, Venus, we have identified many toxic sequences - most of them located in the open reading frame of CD95L. We propose that specific toxic RNAi-active sequences present in the genome can kill cancer cells. PMID:29063830

  15. Strong electromagnetic pulses generated in high-intensity laser-matter interactions

    NASA Astrophysics Data System (ADS)

    Rączka, P.; Dubois, J.-L.; Hulin, S.; Rosiński, M.; Zaraś-Szydłowska, A.; Badziak, J.

    2018-01-01

    Results are reported of an experiment performed at the Eclipse laser facility in CELIA, Bordeaux, on the generation of strong electromagnetic pulses. Measurements were performed of the target neutralization current, the total target charge and the tangential component of the magnetic field for the laser energies ranging from 45 mJ to 92 mJ with the pulse duration approximately 40 fs, and for the pulse durations ranging from 39 fs to 1000 fs, with the laser energy approximately 90 mJ. It was found that the values obtained for thick (mm scale) Cu targets are visibly higher than values reported in previous experiments, which is argued to be a manifestation of a strong dependence of the target electric polarization process on the laser contrast and hence on the amount of preplasma. It was also found that values obtained for thin (μm scale) Al foils were visibly higher than values for thick Cu targets, especially for pulse durations longer than 100 fs. The correlations between the total target charge versus the maximum value of the target neutralization current, and the maximum value of the tangential component of the magnetic field versus the total target charge were analysed. They were found to be in very good agreement with correlations seen in data from previous experiments, which provides a good consistency check on our experimental procedures.

  16. Relativistic Electron Acceleration with Ultrashort Mid-IR Laser Pulses

    NASA Astrophysics Data System (ADS)

    Feder, Linus; Woodbury, Daniel; Shumakova, Valentina; Gollner, Claudia; Miao, Bo; Schwartz, Robert; Pugžlys, Audrius; Baltuška, Andrius; Milchberg, Howard

    2017-10-01

    We report the first results of laser plasma wakefield acceleration driven by ultrashort mid-infrared laser pulses (λ = 3.9 μm , pulsewidth 100 fs, energy <20 mJ, peak power <1 TW), which enables near- and above-critical density interactions with moderate-density gas jets. We present thresholds for electron acceleration based on critical parameters for relativistic self-focusing and target width, as well as trends in the accelerated beam profiles, charge and energy spectra which are supported by 3D particle-in-cell simulations. These results extend earlier work with sub-TW self-modulated laser wakefield acceleration using near IR drivers to the Mid-IR, and enable us to capture time-resolved images of relativistic self-focusing of the laser pulse. This work supported by DOE (DESC0010706TDD, DESC0015516); AFOSR(FA95501310044, FA95501610121); NSF(PHY1535519); DHS.

  17. Enhancement of conversion efficiency of extreme ultraviolet radiation from a liquid aqueous solution microjet target by use of dual laser pulses

    NASA Astrophysics Data System (ADS)

    Higashiguchi, Takeshi; Dojyo, Naoto; Hamada, Masaya; Kawasaki, Keita; Sasaki, Wataru; Kubodera, Shoichi

    2006-03-01

    We demonstrated a debris-free, efficient laser-produced plasma extreme ultraviolet (EUV) source by use of a regenerative liquid microjet target containing tin-dioxide (SnO II) nano-particles. By using a low SnO II concentration (6%) solution and dual laser pulses for the plasma control, we observed the EUV conversion efficiency of 1.2% with undetectable debris.

  18. Note: Pulsed optically pumped atomic clock based on a paraffin-coated cell

    NASA Astrophysics Data System (ADS)

    Lin, Haixiao; Deng, Jianliao; Lin, Jinda; Zhang, Song; Hu, Yao; Wang, Yuzhu

    2018-06-01

    We report on the implementation of a pulsed optically pumped atomic clock based on a paraffin-coated cell. The relaxation times are measured, with the longitudinal relaxation time, T1 = 9.7 ± 0.4 ms, and the transversal relaxation time, T2 = 0.40 ± 0.03 ms. We demonstrated that the measured frequency stability of the clock is 3.9 × 10-13 τ-1/2 (1 s ≤ τ ≤ 100 s) and reaches a value of 3.1 × 10-14 for τ = 1000 s, where τ is the averaging time. This is an unprecedented result for a paraffin-coated vapor cell clock, and it makes significant contributions toward improving the performance of the wall-coated vapor cell atomic clock.

  19. Pulse generation and preamplification for long pulse beamlines of Orion laser facility.

    PubMed

    Hillier, David I; Winter, David N; Hopps, Nicholas W

    2010-06-01

    We describe the pulse generation, shaping, and preamplification system for the nanosecond beamlines of the Orion laser facility. The system generates shaped laser pulses of up to approximately 1 J of 100 ps-5 ns duration with a programmable temporal profile. The laser has a 30th-power supergaussian spatial profile and is diffraction limited. The system is capable of imposing 2D smoothing by spectral dispersion upon the beam, which will produce a nonuniformity of 10% rms at the target.

  20. Rotary target method to prepare thin films of CdS/SiO 2 by pulsed laser deposition

    NASA Astrophysics Data System (ADS)

    Wang, H.; Zhu, Y.; Ong, P. P.

    2000-12-01

    Thin films of CdS-doped SiO 2 glass were prepared by using the conventional pulsed laser deposition (PLD) technique. The laser target consisted of a specially constructed rotary wheel which provided easy control of the exposure-area ratio to expose alternately the two materials to the laser beam. The physical target assembly avoided the potential complications inherent in chemically mixed targets such as in the sol-gel method. Time-of-flight (TOF) spectra confirmed the existence of the SiO 2 and CdS components in the thin-film samples so produced. X-ray diffraction (XRD) and atomic force microscopy(AFM) results showed the different sizes and structures of the as-deposited and annealed films. The wurtzite phase of CdS was found in the 600 oC-annealed sample, while the as-deposited film showed a cubic-hexagonal mixed structure. In the corresponding PL (photoluminescence) spectra, a red shift of the CdS band edge emission was found, which may be a result of the interaction between the CdS nanocrystallite and SiO 2 at their interface.

  1. Radially polarized, half-cycle, attosecond pulses from laser wakefields through coherent synchrotronlike radiation.

    PubMed

    Li, F Y; Sheng, Z M; Chen, M; Yu, L L; Meyer-ter-Vehn, J; Mori, W B; Zhang, J

    2014-10-01

    Attosecond bursts of coherent synchrotronlike radiation are found when driving ultrathin relativistic electron disks in a quasi-one-dimensional regime of wakefield acceleration, in which the laser waist is larger than the wake wavelength. The disks of overcritical density shrink radially due to focusing wakefields, thus providing the transverse currents for the emission of an intense, radially polarized, half-cycle pulse of about 100 attoseconds in duration. The electromagnetic pulse first focuses to a peak intensity (7×10(20)W/cm(2)) 10 times larger than the driving pulse and then emerges as a conical beam. Basic dynamics of the radiative process are derived analytically and in agreement with particle-in-cell simulations. By making use of gas targets instead of solids to form the ultrathin disks, this method allows for high repetition rates required for applications.

  2. Characterizing rapid capacity fade and impedance evolution in high rate pulsed discharged lithium iron phosphate cells for complex, high power loads

    NASA Astrophysics Data System (ADS)

    Wong, Derek N.; Wetz, David A.; Heinzel, John M.; Mansour, Azzam N.

    2016-10-01

    Three 26650 LiFePO4 (LFP) cells are cycled using a 40 A pulsed charge/discharge profile to study their performance in high rate pulsed applications. This profile is used to simulate naval pulsed power loads planned for deployment aboard future vessels. The LFP cells studied experienced an exponential drop in their usable high-rate recharge capacity within sixty cycles due to a rapid rise in their internal resistance. Differential capacitance shows that the voltage window for charge storage is pushed outside of the recommended voltage cutoff limits. Investigation into the state of health of the electrodes shows minimal loss of active material from the cathode to side reactions. Post-mortem examination of the anodic surface films reveals a large increase in the concentration of reduced salt compounds indicating that the pulsed profile creates highly favorable conditions for LiPF6 salt to break down into LiF. This film slows the ionic movement at the interface, affecting transfer kinetics, resulting in charge buildup in the bulk anode without successful energy storage. The results indicate that the use of these cells as a power supply for high pulsed power loads is hindered because of ionically resistant film development and not by an increasing rate of active material loss.

  3. EGFR-targeted granzyme B expressed in NK cells enhances natural cytotoxicity and mediates specific killing of tumor cells.

    PubMed

    Oberoi, Pranav; Jabulowsky, Robert A; Bähr-Mahmud, Hayat; Wels, Winfried S

    2013-01-01

    Natural killer (NK) cells are highly specialized effectors of the innate immune system that hold promise for adoptive cancer immunotherapy. Their cell killing activity is primarily mediated by the pro-apoptotic serine protease granzyme B (GrB), which enters targets cells with the help of the pore-forming protein perforin. We investigated expression of a chimeric GrB fusion protein in NK cells as a means to augment their antitumoral activity. For selective targeting to tumor cells, we fused the epidermal growth factor receptor (EGFR) peptide ligand transforming growth factor α (TGFα) to human pre-pro-GrB. Established human NKL natural killer cells transduced with a lentiviral vector expressed this GrB-TGFα (GrB-T) molecule in amounts comparable to endogenous wildtype GrB. Activation of the genetically modified NK cells by cognate target cells resulted in the release of GrB-T together with endogenous granzymes and perforin, which augmented the effector cells' natural cytotoxicity against NK-sensitive tumor cells. Likewise, GrB-T was released into the extracellular space upon induction of degranulation with PMA and ionomycin. Secreted GrB-T fusion protein displayed specific binding to EGFR-overexpressing tumor cells, enzymatic activity, and selective target cell killing in the presence of an endosomolytic activity. Our data demonstrate that ectopic expression of a targeted GrB fusion protein in NK cells is feasible and can enhance antitumoral activity of the effector cells.

  4. Repetitive output laser system and method using target reflectivity

    DOEpatents

    Johnson, Roy R.

    1978-01-01

    An improved laser system and method for implosion of a thermonuclear fuel pellet in which that portion of a laser pulse reflected by the target pellet is utilized in the laser system to initiate a succeeding target implosion, and in which the energy stored in the laser system to amplify the initial laser pulse, but not completely absorbed thereby, is used to amplify succeeding laser pulses initiated by target reflection.

  5. Effects of the pulsed electromagnetic field PST® on human tendon stem cells: a controlled laboratory study.

    PubMed

    Randelli, Pietro; Menon, Alessandra; Ragone, Vincenza; Creo, Pasquale; Alfieri Montrasio, Umberto; Perucca Orfei, Carlotta; Banfi, Giuseppe; Cabitza, Paolo; Tettamanti, Guido; Anastasia, Luigi

    2016-08-18

    Current clinical procedures for rotator cuff tears need to be improved, as a high rate of failure is still observed. Therefore, new approaches have been attempted to stimulate self-regeneration, including biophysical stimulation modalities, such as low-frequency pulsed electromagnetic fields, which are alternative and non-invasive methods that seem to produce satisfying therapeutic effects. While little is known about their mechanism of action, it has been speculated that they may act on resident stem cells. Thus, the purpose of this study was to evaluate the effects of a pulsed electromagnetic field (PST®) on human tendon stem cells (hTSCs) in order to elucidate the possible mechanism of the observed therapeutic effects. hTSCs from the rotator cuff were isolated from tendon biopsies and cultured in vitro. Then, cells were exposed to a 1-h PST® treatment and compared to control untreated cells in terms of cell morphology, proliferation, viability, migration, and stem cell marker expression. Exposure of hTSCs to PST® did not cause any significant changes in proliferation, viability, migration, and morphology. Instead, while stem cell marker expression significantly decreased in control cells during cell culturing, PST®-treated cells did not have a significant reduction of the same markers. While PST® did not have significant effects on hTSCs proliferation, the treatment had beneficial effects on stem cell marker expression, as treated cells maintained a higher expression of these markers during culturing. These results support the notion that PST® treatment may increase the patient stem cell regenerative potential.

  6. Adoptive therapy with CAR redirected T cells: the challenges in targeting solid tumors.

    PubMed

    Abken, Hinrich

    2015-01-01

    Recent spectacular success in the adoptive cell therapy of leukemia and lymphoma with chimeric antigen receptor (CAR)-modified T cells raised the expectations that this therapy may be efficacious in a wide range of cancer entities. The expectations are based on the predefined specificity of CAR T cells by an antibody-derived binding domain that acts independently of the natural T-cell receptor, recognizes targets independently of presentation by the major histocompatibility complex and allows targeting toward virtually any cell surface antigen. We here discuss that targeting CAR T cells toward solid tumors faces certain circumstances critical for the therapeutic success. Targeting tumor stroma and taking advantage of TRUCK cells, in other words, CAR T cells with inducible release of a transgenic payload, are some strategies envisaged to overcome current limitations in the near future.

  7. Subcellular Biological Effects of Nanosecond Pulsed Electric Fields

    NASA Astrophysics Data System (ADS)

    Kolb, Juergen F.; Stacey, Michael

    Membranes of biological cells can be charged by exposure to pulsed electric fields. After the potential difference across the barrier reaches critical values on the order of 1 V, pores will form. For moderate pulse parameters of duration and amplitude, the effect is limited to the outer cell membrane. With the exposure to nanosecond pulses of several tens of kilovolts per centimeter, a similar effect is also expected for subcellular membranes and structures. Cells will respond to the disruption by different biochemical processes. This offers possibilities for the development of novel medical therapies, the manipulation of cells and microbiological decontamination.

  8. Steering population transfer of the Na2 molecule by an ultrashort pulse train

    NASA Astrophysics Data System (ADS)

    Niu, Dong-Hua; Wang, Shuo; Zhan, Wei-Shen; Tao, Hong-Cai; Wang, Si-Qi

    2018-05-01

    We theoretically investigate the complete population transfer among quantum states of the Na2 molecule using ultrashort pulse trains using the time-dependent wave packet method. The population accumulation of the target state can be steered by controlling the laser parameters, such as the variable pulse pairs, the different pulse widths, the time delays and the repetition period between two contiguous pulses; in particular, the pulse pairs and the pulse widths have a great effect on the population transfer. The calculations show that the ultrashort pulse train is a feasible solution, which can steer the population transfer from the initial state to the target state efficiently with lower peak intensities.

  9. Preservation of protein fluorescence in embedded human dendritic cells for targeted 3D light and electron microscopy

    PubMed Central

    HÖHN, K.; FUCHS, J.; FRÖBER, A.; KIRMSE, R.; GLASS, B.; ANDERS‐ÖSSWEIN, M.; WALTHER, P.; KRÄUSSLICH, H.‐G.

    2015-01-01

    Summary In this study, we present a correlative microscopy workflow to combine detailed 3D fluorescence light microscopy data with ultrastructural information gained by 3D focused ion beam assisted scanning electron microscopy. The workflow is based on an optimized high pressure freezing/freeze substitution protocol that preserves good ultrastructural detail along with retaining the fluorescence signal in the resin embedded specimens. Consequently, cellular structures of interest can readily be identified and imaged by state of the art 3D confocal fluorescence microscopy and are precisely referenced with respect to an imprinted coordinate system on the surface of the resin block. This allows precise guidance of the focused ion beam assisted scanning electron microscopy and limits the volume to be imaged to the structure of interest. This, in turn, minimizes the total acquisition time necessary to conduct the time consuming ultrastructural scanning electron microscope imaging while eliminating the risk to miss parts of the target structure. We illustrate the value of this workflow for targeting virus compartments, which are formed in HIV‐pulsed mature human dendritic cells. PMID:25786567

  10. Personalized targeted therapy for esophageal squamous cell carcinoma

    PubMed Central

    Kang, Xiaozheng; Chen, Keneng; Li, Yicheng; Li, Jianying; D'Amico, Thomas A; Chen, Xiaoxin

    2015-01-01

    Esophageal squamous cell carcinoma continues to heavily burden clinicians worldwide. Researchers have discovered the genomic landscape of esophageal squamous cell carcinoma, which holds promise for an era of personalized oncology care. One of the most pressing problems facing this issue is to improve the understanding of the newly available genomic data, and identify the driver-gene mutations, pathways, and networks. The emergence of a legion of novel targeted agents has generated much hope and hype regarding more potent treatment regimens, but the accuracy of drug selection is still arguable. Other problems, such as cancer heterogeneity, drug resistance, exceptional responders, and side effects, have to be surmounted. Evolving topics in personalized oncology, such as interpretation of genomics data, issues in targeted therapy, research approaches for targeted therapy, and future perspectives, will be discussed in this editorial. PMID:26167067

  11. IT-25DEVELOPMENTALLY REGULATED ANTIGENS FOR IMMUNOLOGIC TARGETING OF MEDULLOBLASTOMA SUBTYPES

    PubMed Central

    Pham, Christina; Flores, Catherine; Pei, Yanxin; Wechsler-Reya, Robert; Mitchell, Duane

    2014-01-01

    INTRODUCTION: Medulloblastoma (MB) remains incurable in one third of patients despite aggressive multi-modality standard therapies. Immunotherapy presents a promising alternative by specifically targeting cancer cells. To date, there have been no successful immunologic applications targeting MB. Emerging evidence from integrated genomic studies has suggested MB variants arise from deregulation of pathways affecting proliferation of progenitor cell populations within the developing cerebellum. Using total embryonic RNA as a source of tumor rejection antigens is attractive because it can be delivered as a single vaccine, target both known and unknown fetal proteins, and can be refined to preferentially treat distinct MB subtypes. METHODS: We have created two transplantable, syngeneic animal MB models recapitulating human SHH and Group 3 variants to investigate the immunologic targeting of different MB subtypes. We generated T cells specific to the developing mouse cerebellum (P5) and tested their reactivity to target cells pulsed with total RNA from two MB subtypes and the normal brain. Immune responses were evaluated by measuring cytokine secretion following re-stimulation of activated T cells with both normal and tumor cell targets. In vivo antitumor efficacy was also tested in survival studies of intracranial tumor-bearing animals. RESULTS: We generated T cells specific to the developing cerebellum in vitro, confirming the immunogenicity of developmentally regulated antigens. Additionally, we have shown that developmental antigen-specific T cells produce high levels of Th1-type cytokines in response to tumor cells of two immunologically distinct subtypes of MB. Interestingly, developmental antigen specific T cells do not show cross reactivity with the normal brain or subsequent stages of the developing brain after P5. Targeting developmental antigens also conferred a significant survival benefit in a treatment model of Group 3 tumor bearing animals. CONCLUSIONS

  12. Systematic Identification of Combinatorial Drivers and Targets in Cancer Cell Lines

    PubMed Central

    Tabchy, Adel; Eltonsy, Nevine; Housman, David E.; Mills, Gordon B.

    2013-01-01

    There is an urgent need to elicit and validate highly efficacious targets for combinatorial intervention from large scale ongoing molecular characterization efforts of tumors. We established an in silico bioinformatic platform in concert with a high throughput screening platform evaluating 37 novel targeted agents in 669 extensively characterized cancer cell lines reflecting the genomic and tissue-type diversity of human cancers, to systematically identify combinatorial biomarkers of response and co-actionable targets in cancer. Genomic biomarkers discovered in a 141 cell line training set were validated in an independent 359 cell line test set. We identified co-occurring and mutually exclusive genomic events that represent potential drivers and combinatorial targets in cancer. We demonstrate multiple cooperating genomic events that predict sensitivity to drug intervention independent of tumor lineage. The coupling of scalable in silico and biologic high throughput cancer cell line platforms for the identification of co-events in cancer delivers rational combinatorial targets for synthetic lethal approaches with a high potential to pre-empt the emergence of resistance. PMID:23577104

  13. Systematic identification of combinatorial drivers and targets in cancer cell lines.

    PubMed

    Tabchy, Adel; Eltonsy, Nevine; Housman, David E; Mills, Gordon B

    2013-01-01

    There is an urgent need to elicit and validate highly efficacious targets for combinatorial intervention from large scale ongoing molecular characterization efforts of tumors. We established an in silico bioinformatic platform in concert with a high throughput screening platform evaluating 37 novel targeted agents in 669 extensively characterized cancer cell lines reflecting the genomic and tissue-type diversity of human cancers, to systematically identify combinatorial biomarkers of response and co-actionable targets in cancer. Genomic biomarkers discovered in a 141 cell line training set were validated in an independent 359 cell line test set. We identified co-occurring and mutually exclusive genomic events that represent potential drivers and combinatorial targets in cancer. We demonstrate multiple cooperating genomic events that predict sensitivity to drug intervention independent of tumor lineage. The coupling of scalable in silico and biologic high throughput cancer cell line platforms for the identification of co-events in cancer delivers rational combinatorial targets for synthetic lethal approaches with a high potential to pre-empt the emergence of resistance.

  14. Mannan-MUC1-pulsed dendritic cell immunotherapy: a phase I trial in patients with adenocarcinoma.

    PubMed

    Loveland, Bruce E; Zhao, Anne; White, Shane; Gan, Hui; Hamilton, Kate; Xing, Pei-Xiang; Pietersz, Geoffrey A; Apostolopoulos, Vasso; Vaughan, Hilary; Karanikas, Vaios; Kyriakou, Peter; McKenzie, Ian F C; Mitchell, Paul L R

    2006-02-01

    Tumor antigen-loaded dendritic cells show promise for cancer immunotherapy. This phase I study evaluated immunization with autologous dendritic cells pulsed with mannan-MUC1 fusion protein (MFP) to treat patients with advanced malignancy. Eligible patients had adenocarcinoma expressing MUC1, were of performance status 0 to 1, with no autoimmune disease. Patients underwent leukapheresis to generate dendritic cells by culture ex vivo with granulocyte macrophage colony-stimulating factor and interleukin 4 for 5 days. Dendritic cells were then pulsed overnight with MFP and harvested for reinjection. Patients underwent three cycles of leukapheresis and reinjection at monthly intervals. Patients with clinical benefit were able to continue with dendritic cell-MFP immunotherapy. Ten patients with a range of tumor types were enrolled, with median age of 60 years (range, 33-70 years); eight patients were of performance status 0 and two of performance status 1. Dendritic cell-MFP therapy led to strong T-cell IFNgamma Elispot responses to the vaccine and delayed-type hypersensitivity responses at injection sites in nine patients who completed treatments. Immune responses were sustained at 1 year in monitored patients. Antibody responses were seen in three patients only and were of low titer. Side effects were grade 1 only. Two patients with clearly progressive disease (ovarian and renal carcinoma) at entry were stable after initial therapy and went on to further leukapheresis and dendritic cell-MFP immunotherapy. These two patients have now each completed over 3 years of treatment. Immunization produced T-cell responses in all patients with evidence of tumor stabilization in 2 of the 10 advanced cancer patients treated. These data support further clinical evaluation of this dendritic cell-MFP immunotherapy.

  15. Microchimeric cells in systemic lupus erythematosus: targets or innocent bystanders?

    PubMed

    Stevens, A M

    2006-01-01

    During pregnancy maternal and fetal cells commute back and forth leading to fetal microchimerism in the mother and maternal microchimerism in the child that can persist for years after the birth. Chimeric fetal and maternal cells can be hematopoietic or can differentiate into somatic cells in multiple organs, potentially acting as targets for 'autoimmunity' and so have been implicated in the pathogenesis of autoimmune diseases that resemble graft-versus-host disease after stem cell transplantation. Fetal cells have been found in women with systemic lupus erythematosus, both in the blood and a target organ, the kidney, suggesting that they may be involved in pathogenesis. Future studies will address how the host immune system normally tolerates maternal and fetal cells or how the balance may change during autoimmunity.

  16. Free Extracellular miRNA Functionally Targets Cells by Transfecting Exosomes from Their Companion Cells.

    PubMed

    Bryniarski, Krzysztof; Ptak, Wlodzimierz; Martin, Emilia; Nazimek, Katarzyna; Szczepanik, Marian; Sanak, Marek; Askenase, Philip W

    2015-01-01

    Lymph node and spleen cells of mice doubly immunized by epicutaneous and intravenous hapten application produce a suppressive component that inhibits the action of the effector T cells that mediate contact sensitivity reactions. We recently re-investigated this phenomenon in an immunological system. CD8+ T lymphocyte-derived exosomes transferred suppressive miR-150 to the effector T cells antigen-specifically due to exosome surface coat of antibody light chains made by B1a lymphocytes. Extracellular RNA (exRNA) is protected from plasma RNases by carriage in exosomes or by chaperones. Exosome transfer of functional RNA to target cells is well described, whereas the mechanism of transfer of exRNA free of exosomes remains unclear. In the current study we describe extracellular miR-150, extracted from exosomes, yet still able to mediate antigen-specific suppression. We have determined that this was due to miR-150 association with antibody-coated exosomes produced by B1a cell companions of the effector T cells, which resulted in antigen-specific suppression of their function. Thus functional cell targeting by free exRNA can proceed by transfecting companion cell exosomes that then transfer RNA cargo to the acceptor cells. This contrasts with the classical view on release of RNA-containing exosomes from the multivesicular bodies for subsequent intercellular targeting. This new alternate pathway for transfer of exRNA between cells has distinct biological and immunological significance, and since most human blood exRNA is not in exosomes may be relevant to evaluation and treatment of diseases.

  17. Free Extracellular miRNA Functionally Targets Cells by Transfecting Exosomes from Their Companion Cells

    PubMed Central

    Bryniarski, Krzysztof; Ptak, Wlodzimierz; Martin, Emilia; Nazimek, Katarzyna; Szczepanik, Marian; Sanak, Marek; Askenase, Philip W.

    2015-01-01

    Lymph node and spleen cells of mice doubly immunized by epicutaneous and intravenous hapten application produce a suppressive component that inhibits the action of the effector T cells that mediate contact sensitivity reactions. We recently re-investigated this phenomenon in an immunological system. CD8+ T lymphocyte-derived exosomes transferred suppressive miR-150 to the effector T cells antigen-specifically due to exosome surface coat of antibody light chains made by B1a lymphocytes. Extracellular RNA (exRNA) is protected from plasma RNases by carriage in exosomes or by chaperones. Exosome transfer of functional RNA to target cells is well described, whereas the mechanism of transfer of exRNA free of exosomes remains unclear. In the current study we describe extracellular miR-150, extracted from exosomes, yet still able to mediate antigen-specific suppression. We have determined that this was due to miR-150 association with antibody-coated exosomes produced by B1a cell companions of the effector T cells, which resulted in antigen-specific suppression of their function. Thus functional cell targeting by free exRNA can proceed by transfecting companion cell exosomes that then transfer RNA cargo to the acceptor cells. This contrasts with the classical view on release of RNA-containing exosomes from the multivesicular bodies for subsequent intercellular targeting. This new alternate pathway for transfer of exRNA between cells has distinct biological and immunological significance, and since most human blood exRNA is not in exosomes may be relevant to evaluation and treatment of diseases. PMID:25923429

  18. Adoptive immunotherapy for B-cell malignancies with autologous chimeric antigen receptor modified tumor targeted T cells.

    PubMed

    Park, Jae H; Brentjens, Renier J

    2010-04-01

    Chemotherapy-resistant B-cell hematologic malignancies may be cured with allogeneic hematopoietic stem cell transplantation (HSCT), demonstrating the potential susceptibility of these tumors to donor T-cell mediated immune responses. However, high rates of transplant-related morbidity and mortality limit this approach. For this reason, there is an urgent need for less-toxic forms of immune-based cellular therapy to treat these malignancies. Adoptive transfer of autologous T cells genetically modified to express chimeric antigen receptors (CARs) targeted to specific tumor-associated antigens represents an attractive means of overcoming the limitations of conventional HSCT. To this end, investigators have generated CARs targeted to various antigens expressed by B-cell malignancies, optimized the design of these CARs to enhance receptor mediated T cell signaling, and demonstrated significant anti-tumor efficacy of the resulting CAR modified T cells both in vitro and in vivo mouse tumor models. These encouraging preclinical data have justified the translation of this approach to the clinical setting with currently 12 open clinical trials and one completed clinical trial treating various B-cell malignancies utilizing CAR modified T cells targeted to either the CD19 or CD20 B-cell specific antigens.

  19. Vesicle-associated membrane protein 7 (VAMP-7) is essential for target cell killing in a natural killer cell line

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    Marcet-Palacios, Marcelo; Odemuyiwa, Solomon O.; Coughlin, Jason J.

    2008-02-15

    Natural killer cells recognize and induce apoptosis in foreign, transformed or virus-infected cells through the release of perforin and granzymes from secretory lysosomes. Clinically, NK-cell mediated killing is a major limitation to successful allo- and xenotransplantation. The molecular mechanisms that regulate the fusion of granzyme B-containing secretory lysosomes to the plasma membrane in activated NK cells, prior to target cell killing, are not fully understood. Using the NK cell line YT-Indy as a model, we have investigated the expression of SNAP REceptors (SNAREs), both target (t-) and vesicular (v-) SNAREs, and their function in granzyme B-mediated target cell killing. Ourmore » data showed that YT-Indy cells express VAMP-7 and SNAP-23, but not VAMP-2. VAMP-7 was associated with granzyme B-containing lysosomal granules. Using VAMP-7 small interfering RNA (siRNA), we successfully knocked down the expression of VAMP-7 protein in YT-Indy to less than 10% of untreated cells in 24 h. VAMP7-deficient YT-Indy cells activated via co-culture with Jurkat cells released <1 ng/mL of granzyme B, compared to 1.5-2.5 {mu}g/mL from controls. Using Jurkat cells as targets, we showed a 7-fold reduction in NK cell-mediated killing by VAMP-7 deficient YT-Indy cells. Our results show that VAMP-7 is a crucial component of granzyme B release and target cell killing in the NK cell line YT-Indy. Thus, targeting VAMP-7 expression specifically with siRNA, following transplantation, may be a viable strategy for preventing NK cell-mediated transplant rejection, in vivo.« less

  20. Tumor-targeting delivery of herb-based drugs with cell-penetrating/tumor-targeting peptide-modified nanocarriers

    PubMed Central

    Kebebe, Dereje; Liu, Yuanyuan; Wu, Yumei; Vilakhamxay, Maikhone; Liu, Zhidong; Li, Jiawei

    2018-01-01

    Cancer has become one of the leading causes of mortality globally. The major challenges of conventional cancer therapy are the failure of most chemotherapeutic agents to accumulate selectively in tumor cells and their severe systemic side effects. In the past three decades, a number of drug delivery approaches have been discovered to overwhelm the obstacles. Among these, nanocarriers have gained much attention for their excellent and efficient drug delivery systems to improve specific tissue/organ/cell targeting. In order to enhance targeting efficiency further and reduce limitations of nanocarriers, nanoparticle surfaces are functionalized with different ligands. Several kinds of ligand-modified nanomedicines have been reported. Cell-penetrating peptides (CPPs) are promising ligands, attracting the attention of researchers due to their efficiency to transport bioactive molecules intracellularly. However, their lack of specificity and in vivo degradation led to the development of newer types of CPP. Currently, activable CPP and tumor-targeting peptide (TTP)-modified nanocarriers have shown dramatically superior cellular specific uptake, cytotoxicity, and tumor growth inhibition. In this review, we discuss recent advances in tumor-targeting strategies using CPPs and their limitations in tumor delivery systems. Special emphasis is given to activable CPPs and TTPs. Finally, we address the application of CPPs and/or TTPs in the delivery of plant-derived chemotherapeutic agents. PMID:29563797

  1. Deep magnetic capture of magnetically loaded cells for spatially targeted therapeutics.

    PubMed

    Huang, Zheyong; Pei, Ning; Wang, Yanyan; Xie, Xinxing; Sun, Aijun; Shen, Li; Zhang, Shuning; Liu, Xuebo; Zou, Yunzeng; Qian, Juying; Ge, Junbo

    2010-03-01

    Magnetic targeting has recently demonstrated potential in promoting magnetically loaded cell delivery to target lesion, but its application is limited by magnetic attenuation. For deep magnetic capture of cells for spatial targeting therapeutics, we designed a magnetic pole, in which the magnetic field density can be focused at a distance from the pole. As flowing through a tube served as a model of blood vessels, the magnetically loaded mesenchymal stem cells (MagMSCs) were highly enriched at the site distance from the magnetic pole. The cell capture efficiency was positively influenced by the magnetic flux density, and inversely influenced by the flow velocity, and well-fitted with the deductive value by theoretical considerations. It appeared to us that the spatially-focused property of the magnetic apparatus promises a new deep targeting strategy to promote homing and engraftment for cellular therapy. Copyright (c) 2009 Elsevier Ltd. All rights reserved.

  2. Controversies in cancer stem cells: targeting embryonic signaling pathways.

    PubMed

    Takebe, Naoko; Ivy, S Percy

    2010-06-15

    Selectively targeting cancer stem cells (CSC) or tumor-initiating cells (TIC; from this point onward referred to as CSCs) with novel agents is a rapidly emerging field of oncology. Our knowledge of CSCs and their niche microenvironments remains a nascent field. CSC's critical dependence upon self-renewal makes these regulatory signaling pathways ripe for the development of experimental therapeutic agents. Investigational agents targeting the Notch, Hedgehog, and Wnt pathways are currently in late preclinical development stages, with some early phase 1-2 testing in human subjects. This series of articles will provide an overview and summary of the current state of knowledge of CSCs, their interactive microenvironment, and how they may serve as important targets for antitumor therapies. We also examine the scope and stage of development of early experimental agents that specifically target these highly conserved embryonic signaling pathways. (c) 2010 AACR.

  3. Pulse electro-deposition of copper on molybdenum for Cu(In,Ga)Se2 and Cu2ZnSnSe4 solar cell applications

    NASA Astrophysics Data System (ADS)

    Bi, Jinlian; Yao, Liyong; Ao, Jianping; Gao, Shoushuai; Sun, Guozhong; He, Qing; Zhou, Zhiqiang; Sun, Yun; Zhang, Yi

    2016-09-01

    The issues of rough surface morphology and the incorporated additives of the electro-deposited Cu layers, which exists in electrodeposition-based processes, is one of the major obstacles to improve the efficiency of Cu(In,Ga)Se2 (CIGSe) and Cu2ZnSnSe4 (CZTSe) solar cells. In this study, the pulse current electro-deposition method is employed to deposit smooth Cu film on Mo substrate in CuSO4 solution without any additives. Grain size of the deposited Cu film is decreased by high cathode polarization successfully. And the concentration polarization, which results from high pulse current density, is controlled successfully by adjusting the pulse frequency. Flat Cu film with smooth surface and compact structure is deposited as pulse current density @ 62.5 mA cm-2, pulse frequency @100,000 Hz, and duty cycle @ 25%. CIGSe and CZTSe absorber films with flat surface and uniform elemental distribution are prepared by selenizing the stacking metal layers electro-deposited by pulse current method. Finally, the CIGSe and CZTSe solar cells with conversion efficiency of 10.39% and 7.83% respectively are fabricated based on the smooth Cu films, which are better than the solar cells fabricated by the rough Cu film deposited by direct current electro-deposition method.

  4. MPN estimation of qPCR target sequence recoveries from whole cell calibrator samples.

    PubMed

    Sivaganesan, Mano; Siefring, Shawn; Varma, Manju; Haugland, Richard A

    2011-12-01

    DNA extracts from enumerated target organism cells (calibrator samples) have been used for estimating Enterococcus cell equivalent densities in surface waters by a comparative cycle threshold (Ct) qPCR analysis method. To compare surface water Enterococcus density estimates from different studies by this approach, either a consistent source of calibrator cells must be used or the estimates must account for any differences in target sequence recoveries from different sources of calibrator cells. In this report we describe two methods for estimating target sequence recoveries from whole cell calibrator samples based on qPCR analyses of their serially diluted DNA extracts and most probable number (MPN) calculation. The first method employed a traditional MPN calculation approach. The second method employed a Bayesian hierarchical statistical modeling approach and a Monte Carlo Markov Chain (MCMC) simulation method to account for the uncertainty in these estimates associated with different individual samples of the cell preparations, different dilutions of the DNA extracts and different qPCR analytical runs. The two methods were applied to estimate mean target sequence recoveries per cell from two different lots of a commercially available source of enumerated Enterococcus cell preparations. The mean target sequence recovery estimates (and standard errors) per cell from Lot A and B cell preparations by the Bayesian method were 22.73 (3.4) and 11.76 (2.4), respectively, when the data were adjusted for potential false positive results. Means were similar for the traditional MPN approach which cannot comparably assess uncertainty in the estimates. Cell numbers and estimates of recoverable target sequences in calibrator samples prepared from the two cell sources were also used to estimate cell equivalent and target sequence quantities recovered from surface water samples in a comparative Ct method. Our results illustrate the utility of the Bayesian method in accounting for

  5. Feedback Interactions of Polymerized Actin with the Cell Membrane: Waves, Pulses, and Oscillations

    NASA Astrophysics Data System (ADS)

    Carlsson, Anders

    Polymerized filaments of the protein actin have crucial functions in cell migration, and in bending the cell membrane to drive endocytosis or the formation of protrusions. The nucleation and polymerization of actin filaments are controlled by upstream agents in the cell membrane, including nucleation-promoting factors (NPFs) that activate the Arp2/3 complex to form new branches on pre-existing filaments. But polymerized actin (F-actin) also feeds back on the assembly of NPFs. We explore the effects of the resulting feedback loop of F-actin and NPFs on two phenomena: actin pulses that drive endocytosis in yeast, and actin waves traveling along the membrane of several cell types. In our model of endocytosis in yeast, the actin network is grown explicitly in three dimensions, exerts a negative feedback interaction on localized patch of NPFs in the membrane, and bends the membrane by exerting a distribution of forces. This model explains observed actin and NPF pulse dynamics, and the effects of several interventions including i) NPF mutations, ii) inhibition of actin polymerization, and iii) deletion of a protein that allows F-actin to bend the cell membrane. The model predicts that mutation of the active region of an NPF will enhance the accumulation of that NPF, and we confirm this prediction by quantitative fluorescence microscopy. For actin waves, we treat a similar model, with NPFs distributed over a larger region of the cell membrane. This model naturally generates actin waves, and predicts a transition from wave behavior to spatially localized oscillations when NPFs are confined to a small region. We also predict a transition from waves to static polarization as the negative-feedback coupling between F-actin and the NPFs is reduced. Supported by NIGMS Grant R01 GM107667.

  6. ERK/p38 MAPK inhibition reduces radio-resistance to a pulsed proton beam in breast cancer stem cells

    NASA Astrophysics Data System (ADS)

    Jung, Myung-Hwan; Park, Jeong Chan

    2015-10-01

    Recent studies have identified highly tumorigenic cells with stem cell-like characteristics, termed cancer stem cells (CSCs) in human cancers. CSCs are resistant to conventional radiotherapy and chemotherapy owing to their high DNA repair ability and oncogene overexpression. However, the mechanisms regulating CSC radio-resistance, particularly proton beam resistance, remain unclear. We isolated CSCs from the breast cancer cell lines MCF-7 and MDA-MB-231, which expressed the characteristic breast CSC membrane protein markers CD44+/CD24-/ low , and irradiated the CSCs with pulsed proton beams. We confirmed that CSCs were resistant to pulsed proton beams and showed that treatment with p38 and ERK inhibitors reduced CSC radio-resistance. Based on these results, BCSC radio-resistance can be reduced during proton beam therapy by co-treatment with ERK1/2 or p38 inhibitors, a novel approach to breast cancer therapy.

  7. Ultrafast laser ablation for targeted atherosclerotic plaque removal

    NASA Astrophysics Data System (ADS)

    Lanvin, Thomas; Conkey, Donald B.; Descloux, Laurent; Frobert, Aurelien; Valentin, Jeremy; Goy, Jean-Jacques; Cook, Stéphane; Giraud, Marie-Noelle; Psaltis, Demetri

    2015-07-01

    Coronary artery disease, the main cause of heart disease, develops as immune cells and lipids accumulate into plaques within the coronary arterial wall. As a plaque grows, the tissue layer (fibrous cap) separating it from the blood flow becomes thinner and increasingly susceptible to rupturing and causing a potentially lethal thrombosis. The stabilization and/or treatment of atherosclerotic plaque is required to prevent rupturing and remains an unsolved medical problem. Here we show for the first time targeted, subsurface ablation of atherosclerotic plaque using ultrafast laser pulses. Excised atherosclerotic mouse aortas were ablated with ultrafast near-infrared (NIR) laser pulses. The physical damage was characterized with histological sections of the ablated atherosclerotic arteries from six different mice. The ultrafast ablation system was integrated with optical coherence tomography (OCT) imaging for plaque-specific targeting and monitoring of the resulting ablation volume. We find that ultrafast ablation of plaque just below the surface is possible without causing damage to the fibrous cap, which indicates the potential use of ultrafast ablation for subsurface atherosclerotic plaque removal. We further demonstrate ex vivo subsurface ablation of a plaque volume through a catheter device with the high-energy ultrafast pulse delivered via hollow-core photonic crystal fiber.

  8. Pulsed Electromagnetic Field Assisted in vitro Electroporation: A Pilot Study

    NASA Astrophysics Data System (ADS)

    Novickij, Vitalij; Grainys, Audrius; Lastauskienė, Eglė; Kananavičiūtė, Rūta; Pamedytytė, Dovilė; Kalėdienė, Lilija; Novickij, Jurij; Miklavčič, Damijan

    2016-09-01

    Electroporation is a phenomenon occurring due to exposure of cells to Pulsed Electric Fields (PEF) which leads to increase of membrane permeability. Electroporation is used in medicine, biotechnology, and food processing. Recently, as an alternative to electroporation by PEF, Pulsed ElectroMagnetic Fields (PEMF) application causing similar biological effects was suggested. Since induced electric field in PEMF however is 2-3 magnitudes lower than in PEF electroporation, the membrane permeabilization mechanism remains hypothetical. We have designed pilot experiments where Saccharomyces cerevisiae and Candida lusitaniae cells were subjected to single 100-250 μs electrical pulse of 800 V with and without concomitant delivery of magnetic pulse (3, 6 and 9 T). As expected, after the PEF pulses only the number of Propidium Iodide (PI) fluorescent cells has increased, indicative of membrane permeabilization. We further show that single sub-millisecond magnetic field pulse did not cause detectable poration of yeast. Concomitant exposure of cells to pulsed electric (PEF) and magnetic field (PMF) however resulted in the increased number PI fluorescent cells and reduced viability. Our results show increased membrane permeability by PEF when combined with magnetic field pulse, which can explain electroporation at considerably lower electric field strengths induced by PEMF compared to classical electroporation.

  9. Pulsed Electromagnetic Field Assisted in vitro Electroporation: A Pilot Study

    PubMed Central

    Novickij, Vitalij; Grainys, Audrius; Lastauskienė, Eglė; Kananavičiūtė, Rūta; Pamedytytė, Dovilė; Kalėdienė, Lilija; Novickij, Jurij; Miklavčič, Damijan

    2016-01-01

    Electroporation is a phenomenon occurring due to exposure of cells to Pulsed Electric Fields (PEF) which leads to increase of membrane permeability. Electroporation is used in medicine, biotechnology, and food processing. Recently, as an alternative to electroporation by PEF, Pulsed ElectroMagnetic Fields (PEMF) application causing similar biological effects was suggested. Since induced electric field in PEMF however is 2–3 magnitudes lower than in PEF electroporation, the membrane permeabilization mechanism remains hypothetical. We have designed pilot experiments where Saccharomyces cerevisiae and Candida lusitaniae cells were subjected to single 100–250 μs electrical pulse of 800 V with and without concomitant delivery of magnetic pulse (3, 6 and 9 T). As expected, after the PEF pulses only the number of Propidium Iodide (PI) fluorescent cells has increased, indicative of membrane permeabilization. We further show that single sub-millisecond magnetic field pulse did not cause detectable poration of yeast. Concomitant exposure of cells to pulsed electric (PEF) and magnetic field (PMF) however resulted in the increased number PI fluorescent cells and reduced viability. Our results show increased membrane permeability by PEF when combined with magnetic field pulse, which can explain electroporation at considerably lower electric field strengths induced by PEMF compared to classical electroporation. PMID:27634482

  10. Liver cell-targeted delivery of therapeutic molecules.

    PubMed

    Kang, Jeong-Hun; Toita, Riki; Murata, Masaharu

    2016-01-01

    The liver is the largest internal organ in mammals and is involved in metabolism, detoxification, synthesis of proteins and lipids, secretion of cytokines and growth factors and immune/inflammatory responses. Hepatitis, alcoholic or non-alcoholic liver disease, hepatocellular carcinoma, hepatic veno-occlusive disease, and liver fibrosis and cirrhosis are the most common liver diseases. Safe and efficient delivery of therapeutic molecules (drugs, genes or proteins) into the liver is very important to increase the clinical efficacy of these molecules and to reduce their side effects in other organs. Several liver cell-targeted delivery systems have been developed and tested in vivo or ex vivo/in vitro. In this review, we discuss the literature concerning liver cell-targeted delivery systems, with a particular emphasis on the results of in vivo studies.

  11. Metabolic and structural integrity of magnetic nanoparticle-loaded primary endothelial cells for targeted cell therapy.

    PubMed

    Orynbayeva, Zulfiya; Sensenig, Richard; Polyak, Boris

    2015-05-01

    To successfully translate magnetically mediated cell targeting from bench to bedside, there is a need to systematically assess the potential adverse effects of magnetic nanoparticles (MNPs) interacting with 'therapeutic' cells. Here, we examined in detail the effects of internalized polymeric MNPs on primary rat endothelial cells' structural intactness, metabolic integrity and proliferation potential. The intactness of cytoskeleton and organelles was studied by fluorescent confocal microscopy, flow cytometry and high-resolution respirometry. MNP-loaded primary endothelial cells preserve intact cytoskeleton and organelles, maintain normal rate of proliferation, calcium signaling and mitochondria energy metabolism. This study provides supportive evidence that MNPs at doses necessary for targeting did not induce significant adverse effects on structural integrity and functionality of primary endothelial cells - potential cell therapy vectors.

  12. Cooperative pulses for pseudo-pure state preparation

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    Wei, Daxiu; Chang, Yan; Yang, Xiaodong, E-mail: steffen.glaser@tum.de, E-mail: xiaodong.yang@sibet.ac.cn

    2014-06-16

    Using an extended version of the optimal-control-based gradient ascent pulse engineering algorithm, cooperative (COOP) pulses are designed for multi-scan experiments to prepare pseudo-pure states in quantum computation. COOP pulses can cancel undesired signal contributions, complementing and generalizing phase cycles. They also provide more flexibility and, in particular, eliminate the need to select specific individual target states and achieve the fidelity of theoretical limit by flexibly choosing appropriate number of scans and duration of pulses. The COOP approach is experimentally demonstrated for three-qubit and four-qubit systems.

  13. Nanoparticle mediated ablation of breast cancer cells using a nanosecond pulsed electric field

    NASA Astrophysics Data System (ADS)

    Burford, Christopher

    In the past, both nanomaterials and various heating modalities have been researched as means for treating cancers. However, many of the current methodologies have the flaws of inconsistent tumor ablation and significant destruction of healthy cells. Based on research performed using constant radiofrequency electric fields and metallic nanoparticles (where cell necrosis is induced by the heating of these nanoparticles) we have developed a modality that simlarly uses functionalized metallic nanoparticles, specific for the T47D breast cancer cell line, and nanosecond pulsed electric fields as the hyperthermic inducer. Using both iron oxide and gold nanoparticles the results of our pilot studies indicated that up to 90% of the cancer cells were ablated given the optimal treatment parameters. These quantities of ablated cells were achieved using a cumulative exposure time 6 orders of magnitude less than most in vitro radiofrequency electric field studies.

  14. EUV emission stimulated by use of dual laser pulses from continus liquid microjet targets

    NASA Astrophysics Data System (ADS)

    Higashiguchi, Takeshi; Rajyaguru, Chirag; Sasaki, Wataru; Kubodera, Shoichi

    2004-11-01

    A continuous water-jet or water-jet mixed with LiF with several tens μm diameter was formed in a vacuum chamber through a small capillary nozzle. Usage of two laser pulses is an efficient way to produce EUV emission, since a density and temperature of a plasma formed by the first laser pulse are regulated by the second laser pulse. By adjusting the delay of the second pulse, one could maximize the EUV emission. A subpicosecond Ti:Sapphire laser at a wavelength of 800 nm produced a maximum energy around 30 mJ. The beam was divided by a Michelson interferometer, which produced two laser pulses with energies of 5 mJ. The pulse duration was adjusted around 300 fs (FWHM). Both beams were focused on a micro-jet using a lens with a focal length of 15 cm. The delay time between the two pulses was varied from 100 to 800 ps by use of an optical delay line. Clear enhancement of the EUV emission yield was observed when the delay between the two pulses was around 500 ps. The experimentally observed delay agrees reasonably well with that of a plasma to expand to its critical density of 10^21 cm-3.

  15. Irradiation of amelanotic melanoma cells with 532 nm high peak power pulsed laser radiation in the presence of the photothermal sensitizer Cu(II)-hematoporphyrin: a new approach to cell photoinactivation.

    PubMed

    Soncin, M; Busetti, A; Fusi, F; Jori, G; Rodgers, M A

    1999-06-01

    Cu(II)-hematoporphyrin (CuHp) was efficiently accumulated by B78H1 amelanotic melanoma cells upon incubation with porphyrin concentrations up to 52 microM. When the cells incubated for 18 h with 13 microM CuHp were irradiated with 532 nm light from a Q-switched Nd: YAG laser operated in a pulsed mode (10 ns pulses, 10 Hz) a significant decrease in cell survival was observed. The cell photoinactivation was not the consequence of a photodynamic process, as CuHp gave no detectable triplet signal upon laser flash photolysis excitation and no decrease in cell survival was observed upon continuous wave irradiation. Thus, it is likely that CuHp sensitization takes place by photothermal pathways. The efficiency of the photoprocess was modulated by different parameters; thus, while varying the amount of added CuHp in the 3.25-26 microM range had little effect, pulse energies larger than 50 mJ and irradiation times of at least 120 s were necessary to induce a cell inactivation of about 50%. The porphyrin-cell incubation time prior to irradiation had a major influence on cell survival, suggesting that the nature of the CuHp microenvironment can control the efficiency of photothermal sensitization.

  16. A 20-Amino Acid Module of Protein Kinase Cϵ Involved in Translocation and Selective Targeting at Cell-Cell Contacts*

    PubMed Central

    Diouf, Barthélémy; Collazos, Alejandra; Labesse, Gilles; Macari, Françoise; Choquet, Armelle; Clair, Philippe; Gauthier-Rouvière, Cécile; Guérineau, Nathalie C.; Jay, Philippe; Hollande, Frédéric; Joubert, Dominique

    2009-01-01

    In the pituitary gland, activated protein kinase C (PKC) isoforms accumulate either selectively at the cell-cell contact (α and ϵ) or at the entire plasma membrane (β1 and δ). The molecular mechanisms underlying these various subcellular locations are not known. Here, we demonstrate the existence within PKCϵ of a cell-cell contact targeting sequence (3CTS) that, upon stimulation, is capable of targeting PKCδ, chimerin-α1, and the PKCϵ C1 domain to the cell-cell contact. We show that this selective targeting of PKCϵ is lost upon overexpression of 3CTS fused to a (R-Ahx-R)4 (where Ahx is 6-aminohexanoic acid) vectorization peptide, reflecting a dominant-negative effect of the overexpressed 3CTS on targeting selectivity. 3CTS contains a putative amphipathic α-helix, a 14-3-3-binding site, and the Glu-374 amino acid, involved in targeting selectivity. We show that the integrity of the α-helix is important for translocation but that 14-3-3 is not involved in targeting selectivity. However, PKCϵ translocation is increased when PKCϵ/14-3-3 interaction is abolished, suggesting that phorbol 12-myristate 13-acetate activation may initiate two sets of PKCϵ functions, those depending on 14-3-3 and those depending on translocation to cell-cell contacts. Thus, 3CTS is involved in the modulation of translocation via its 14-3-3-binding site, in cytoplasmic desequestration via the α-helix, and in selective PKCϵ targeting at the cell-cell contact via Glu-374. PMID:19429675

  17. E-selectin liposomal and nanotube-targeted delivery of doxorubicin to circulating tumor cells

    PubMed Central

    Mitchell, Michael J.; Chen, Christina S.; Ponmudi, Varun; Hughes, Andrew D.; King, Michael R.

    2012-01-01

    The presence of circulating tumor cells (CTCs) is believed to lead to the formation of secondary tumors via an adhesion cascade involving interaction between adhesion receptors of endothelial cells and ligands on CTCs. Many CTCs express sialylated carbohydrate ligands on their surfaces that adhere to selectin protein found on inflamed endothelial cells. We have investigated the feasibility of using immobilized selectin proteins as a targeting mechanism for CTCs under flow. Herein, targeted liposomal doxorubicin (L-DXR) was functionalized with recombinant human E-selectin (ES) and polyethylene glycol (PEG) to target and kill cancer cells under shear flow, both when immobilized along a microtube device or sheared in a cone-and-plate viscometer in a dilute suspension. Healthy circulating cells such as red blood cells were not targeted by this mechanism and were left to freely circulate, and minimal leukocyte death was observed. Halloysite nanotube (HNT)-coated microtube devices immobilized with nanoscale liposomes significantly enhanced the targeting, capture, and killing of cancer cells. This work demonstrates that E-selectin functionalized L-DXR, sheared in suspension or immobilized onto microtube devices, provides a novel approach to selectively target and deliver chemotherapeutics to CTCs in the bloodstream. PMID:22421423

  18. The effect of frequency doubled double pulse Nd:YAG laser fiber proximity to the target stone on transient cavitation and acoustic emission.

    PubMed

    Fuh, Eric; Haleblian, George E; Norris, Regina D; Albala, W David M; Simmons, Neal; Zhong, Pei; Preminger, Glenn M

    2007-04-01

    Scant information has been published describing the effect of laser fiber distance from the stone target on the mechanism of calculus fragmentation. Using high speed photography and acoustic emission measurements we characterized the impact of laser fiber proximity on stone comminution. We evaluated the effect of laser fiber distance from the stone target on resultant cavitation bubble formation and shock wave generation. Stone fragmentation was assessed using a FREDDY (frequency doubled double pulse Nd:YAG) (World of Medicine, Orlando, Florida) laser and a holmium laser. The FREDDY laser was operated using a 420 microm fiber at an output energy of 120 and 160 mJ in single and double pulse settings, and a pulse repetition rate of 1 Hz. The holmium laser was operated using a 200 microm fiber at an output energy of 1 to 3 J and a pulse repetition rate of 1 Hz. The surface of a 1 cm square BegoStone (Bego, Bremen, Germany) attached to an X-Y-Z translational stage was aligned perpendicular to the laser fiber, which was immersed in a Lucite tank filled with water at room temperature. An Imacon 200 high speed camera was used to capture transient cavitation bubbles at a framing rate of up to 1,000,000 frames per second. Acoustic emission signals associated with shock waves generated during the rapid expansion and collapse of the cavitation bubble were measured using a 1 MHz focused ultrasound transducer. At laser fiber distances of 3.0 mm or less cavitation bubbles and shock waves were observed with the FREDDY laser. In contrast to the holmium laser, the bubble size and shock wave intensity of the FREDDY laser was inversely related to the fiber-to-stone distance over the range tested (0.5 to 3.0 mm). While bubble size was noted to increase with a larger stone-to-fiber distance using the holmium laser, to consistently generate cavitation bubbles and shock waves using the FREDDY laser the laser fiber should be operated within 3.0 mm of the target stone. These findings have

  19. Concise Review: Cell Surface N-Linked Glycoproteins as Potential Stem Cell Markers and Drug Targets.

    PubMed

    Boheler, Kenneth R; Gundry, Rebekah L

    2017-01-01

    Stem cells and their derivatives hold great promise to advance regenerative medicine. Critical to the progression of this field is the identification and utilization of antibody-accessible cell-surface proteins for immunophenotyping and cell sorting-techniques essential for assessment and isolation of defined cell populations with known functional and therapeutic properties. Beyond their utility for cell identification and selection, cell-surface proteins are also major targets for pharmacological intervention. Although comprehensive cell-surface protein maps are highly valuable, they have been difficult to define until recently. In this review, we discuss the application of a contemporary targeted chemoproteomic-based technique for defining the cell-surface proteomes of stem and progenitor cells. In applying this approach to pluripotent stem cells (PSCs), these studies have improved the biological understanding of these cells, led to the enhanced use and development of antibodies suitable for immunophenotyping and sorting, and contributed to the repurposing of existing drugs without the need for high-throughput screening. The utility of this latter approach was first demonstrated with human PSCs (hPSCs) through the identification of small molecules that are selectively toxic to hPSCs and have the potential for eliminating confounding and tumorigenic cells in hPSC-derived progeny destined for research and transplantation. Overall, the cutting-edge technologies reviewed here will accelerate the development of novel cell-surface protein targets for immunophenotyping, new reagents to improve the isolation of therapeutically qualified cells, and pharmacological studies to advance the treatment of intractable diseases amenable to cell-replacement therapies. Stem Cells Translational Medicine 2017;6:131-138. © 2016 The Authors Stem Cells Translational Medicine published by Wiley Periodicals, Inc. on behalf of AlphaMed Press.

  20. The effects of pulsed electromagnetic field (PEMF) on osteoblast-like cells cultured on titanium and titanium-zirconium surfaces.

    PubMed

    Atalay, Belir; Aybar, Buket; Ergüven, Mine; Emes, Yusuf; Bultan, Özgür; Akça, Kivanç; Yalçin, Serhat; Baysal, Uğur; Işsever, Halim; Çehreli, Murat Cavit; Bilir, Ayhan

    2013-11-01

    Commercially pure Ti, together with Ti Ni, Ti-6Al-4V, and Ti-6Al-7Nb alloys, are among the materials currently being used for this purpose. Titanium-zirconium (TiZr) has been developed that allows SLActive surface modification and that has comparable or better mechanical strength and improved biocompatibility compared with existing Ti alloys. Furthermore, approaches have targeted making the implant surface more hydrophilic, as with the Straumann SLActive surface, a modification of the SLA surface. The aim of this study is to evaluate the effects of pulsed electromagnetic field (PEMF) to the behavior of neonatal rat calvarial osteoblast-like cells cultured on commercially pure titanium (cpTi) and titanium-zirconium alloy (TiZr) discs with hydrophilic surface properties. Osteoblast cells were cultured on titanium and TiZr discs, and PEMF was applied. Cell proliferation rates, cell numbers, cell viability rates, alkaline phosphatase, and midkine (MK) levels were measured at 24 and 72 hours. At 24 hours, the number of cells was significantly higher in the TiZr group. At 72 hours, TiZr had a significantly higher number of cells when compared to SLActive, SLActive + PEMF, and machine surface + PEMF groups. At 24 hours, cell proliferation was significantly higher in the TiZr group than SLActive and TiZr + PEMF group. At 72 hours, TiZr group had higher proliferation rate than machine surface and TiZr + PEMF. Cell proliferation in the machine surface group was lower than both SLActive + PEMF and machine surface + PEMF. MK levels of PEMF-treated groups were lower than untreated groups for 72 hours. Our findings conclude that TiZr surfaces are similar to cpTi surfaces in terms of biocompatibility. However, PEMF application has a higher stimulative effect on cells cultured on cpTi surfaces when compared to TiZr.