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Sample records for monocrystalline si targets

  1. Surface Damage Mechanism of Monocrystalline Si Under Mechanical Loading

    NASA Astrophysics Data System (ADS)

    Zhao, Qingliang; Zhang, Quanli; To, Suet; Guo, Bing

    2017-03-01

    Single-point diamond scratching and nanoindentation on monocrystalline silicon wafer were performed to investigate the surface damage mechanism of Si under the contact loading. The results showed that three typical stages of material removal appeared during dynamic scratching, and a chemical reaction of Si with the diamond indenter and oxygen occurred under the high temperature. In addition, the Raman spectra of the various points in the scratching groove indicated that the Si-I to β-Sn structure (Si-II) and the following β-Sn structure (Si-II) to amorphous Si transformation appeared under the rapid loading/unloading condition of the diamond grit, and the volume change induced by the phase transformation resulted in a critical depth (ductile-brittle transition) of cut (˜60 nm ± 15 nm) much lower than the theoretical calculated results (˜387 nm). Moreover, it also led to abnormal load-displacement curves in the nanoindentation tests, resulting in the appearance of elbow and pop-out effects (˜270 nm at 20 s, 50 mN), which were highly dependent on the loading/unloading conditions. In summary, phase transformation of Si promoted surface deformation and fracture under both static and dynamic mechanical loading.

  2. The epitaxial growth of (1 1 1) oriented monocrystalline Si film based on a 4:5 Si-to-SiC atomic lattice matching interface

    SciTech Connect

    Yang, Chen; Chen, Zhiming; Hu, Jichao; Ren, Zhanqiang; Lin, Shenghuang

    2012-06-15

    Highlights: ► A monocrystalline Si film was demonstrated by XRD to epitaxially grow on the 6H-SiC substrate. ► A 4:5 Si-to-SiC lattice matching structure was observed at the Si/SiC interface. ► The calculated value of the actual lattice mismatch is only 0.26%. ► Defects can be effectively reduced at the 4:5 Si-to-SiC lattice matching Si/SiC interface. -- Abstract: Due to a huge lattice mismatch of about 20% theoretically existing between SiC and Si, it is difficult for growing monocrystalline Si/SiC heterojunction to realize the light control of SiC devices. However, based on a 4:5 Si-to-SiC atomic lattice matching interface structure, the monocrystalline Si films were epitaxially prepared on the 6H-SiC (0 0 0 1) substrate by hot-wall chemical vapor deposition in our work. The film was characterized by X-ray diffraction analysis with only (1 1 1) orientation occurring. The X-ray rocking curves illustrated good symmetry with a full width at half maximum of 0.4339° omega. A 4:5 Si-to-SiC atomic matching structure of the Si/6H-SiC interface clearly observed by the transmission electron microscope revealed the essence of growing the monocrystalline Si film on the SiC substrate.

  3. Physical assembly of Ag nanocrystals on enclosed surfaces in monocrystalline Si

    PubMed Central

    Martin, Michael S.; Theodore, N. David; Wei, Chao-Chen; Shao, Lin

    2014-01-01

    Growth of thin crystals on external substrate surfaces by many different methods is a well-known technique, but its extension to inner, enclosed surfaces of large defects in monocrystalline materials has not yet been reported. The literature on thin film growth and defects in materials can be leveraged to fabricate new structures for a variety of applications. Here we show a physical process of nucleation and evolution of nanocrystalline silver inside voids in monocrystalline silicon. We found that the Ag growth is hetero-epitaxial using a coincident site lattice. Alignment of Ag and Si atomic planes is uniformly observed by high resolution transmission electron microscopy and macroscopically by channeling Rutherford backscattering spectrometry. PMID:25376502

  4. Nanofabrication on monocrystalline silicon through friction-induced selective etching of Si3N4 mask.

    PubMed

    Guo, Jian; Yu, Bingjun; Wang, Xiaodong; Qian, Linmao

    2014-01-01

    A new fabrication method is proposed to produce nanostructures on monocrystalline silicon based on the friction-induced selective etching of its Si3N4 mask. With low-pressure chemical vapor deposition (LPCVD) Si3N4 film as etching mask on Si(100) surface, the fabrication can be realized by nanoscratching on the Si3N4 mask and post-etching in hydrofluoric acid (HF) and potassium hydroxide (KOH) solution in sequence. Scanning Auger nanoprobe analysis indicated that the HF solution could selectively etch the scratched Si3N4 mask and then provide the gap for post-etching of silicon substrate in KOH solution. Experimental results suggested that the fabrication depth increased with the increase of the scratching load or KOH etching period. Because of the excellent masking ability of the Si3N4 film, the maximum fabrication depth of nanostructure on silicon can reach several microns. Compared to the traditional friction-induced selective etching technique, the present method can fabricate structures with lesser damage and deeper depths. Since the proposed method has been demonstrated to be a less destructive and flexible way to fabricate a large-area texture structure, it will provide new opportunities for Si-based nanofabrication.

  5. Nanofabrication on monocrystalline silicon through friction-induced selective etching of Si3N4 mask

    PubMed Central

    2014-01-01

    A new fabrication method is proposed to produce nanostructures on monocrystalline silicon based on the friction-induced selective etching of its Si3N4 mask. With low-pressure chemical vapor deposition (LPCVD) Si3N4 film as etching mask on Si(100) surface, the fabrication can be realized by nanoscratching on the Si3N4 mask and post-etching in hydrofluoric acid (HF) and potassium hydroxide (KOH) solution in sequence. Scanning Auger nanoprobe analysis indicated that the HF solution could selectively etch the scratched Si3N4 mask and then provide the gap for post-etching of silicon substrate in KOH solution. Experimental results suggested that the fabrication depth increased with the increase of the scratching load or KOH etching period. Because of the excellent masking ability of the Si3N4 film, the maximum fabrication depth of nanostructure on silicon can reach several microns. Compared to the traditional friction-induced selective etching technique, the present method can fabricate structures with lesser damage and deeper depths. Since the proposed method has been demonstrated to be a less destructive and flexible way to fabricate a large-area texture structure, it will provide new opportunities for Si-based nanofabrication. PMID:24940174

  6. Performance Degradation of Encapsulated Monocrystalline-Si Solar Cells upon Accelerated Weathering Exposures: Preprint

    SciTech Connect

    Glick, S. H.; Pern, F. J.; Watson, G. L.; Tomek, D.; Raaff, J.

    2001-10-01

    Presented at 2001 NCPV Program Review Meeting: Performed accelerated exposures to study performance reliability/materials degradation of encapsulated c-Si cells using weathering protocols in 2 weatherometers. We have performed accelerated exposures to study performance reliability and materials degradation of a total of forty-one 3-cm x 3-cm monocrystalline-Si (c-Si) solar cells that were variously encapsulated using accelerated weathering protocols in two weatherometers (WOMs), with and without front specimen water sprays. Laminated cells (EVA/c-Si/EVA, ethylene vinyl acetate) with one of five superstrate/substrate variations and other features including with and without: (i) load resistance, (ii) Al foil light masks, and (iii) epoxy edge-sealing were studied. Three additional samples, omitting EVA, were exposed under a full-spectrum solar simulator, or heated in an oven, for comparison. After exposures, cell performance decreased irregularly, but to a relatively greater extent for samples exposed in WOM where light, heat, and humidity cycles were present (solar simulator or oven lacked such cycles). EVA laminates in the samples masked with aluminum (Al) foils were observed to retain moisture in WOM with water spray. Moisture effects caused substantial efficiency losses probably related in part to increasing series resistance.

  7. Convoy electron production in polycrystalline and monocrystalline targets

    SciTech Connect

    Huldt, S.

    1980-01-01

    The velocity distribution of electrons ejected close to the forward direction by 0.8-2 MeV/A ions traversing various solid targets, including a Au monocrystal, is measured in coincidence with emerging charge-selected ions. The velocity spectrum is observed to be independent of outgoing projectile velocity and charge state for polycrystalline targets. Measurements on the Au crystal under channeling conditions show dependences on final charge state, and are tentatively explained by assuming that the main contribution to the production yield comes from the non-channeled fraction of the ions. A simple model for the creation of the forward-ejected electrons is proposed, which accounts for most of the experimental findings.

  8. Theoretically predicted and experimentally determined effects of the Si/(Si + C) gas phase ratio on the growth and character of monocrystalline beta silicon carbide films

    NASA Astrophysics Data System (ADS)

    Kim, H. J.; Davis, R. F.

    1986-10-01

    The effects of the Si/(Si + C) ratio in the reaction gas stream on the growth and properties of monocrystalline beta-SiC films grown on Si(100) substrates via chemical vapor deposition have been theoretically and experimentally studied. The amounts of condensed phases of beta-SiC and Si, and the partial pressures of the remaining Si and C-containing gases as a function of the Si/(Si + C) ratio in the source gases have been initially obtained from thermodynamic calculations using the 'SOLGASMIX-PV' computer program. Complementary and comparative experimental growth studies have shown that inclusion-free films having maximum values in growth rate and carrier concentration and a minimum value of resistivity were obtained near Si/(Si + C) = 0.5.

  9. Determination of SiO2 and C layers on a monocrystalline silicon sphere by reference-free x-ray fluorescence analysis

    NASA Astrophysics Data System (ADS)

    Hönicke, Philipp; Holfelder, Ina; Kolbe, Michael; Lubeck, Janin; Pollakowski-Herrmann, Beatrix; Unterumsberger, Rainer; Weser, Jan; Beckhoff, Burkhard

    2017-08-01

    The redefinition of the kilogram will be based on Planck’s constant, which can be calculated from Avogadro’s constant, and hence by ‘counting’ the silicon atoms in a monocrystalline 28Si sphere weighing one kilogram. To reduce the influence of the surface layers on the determined value requires, amongst other issues, an accurate quantification of these layers on the 28Si spheres. Apart from the native SiO2 layer, carbonaceous contamination layers and adsorbed water molecules are expected to be crucial in determining the surface composition. The overall mass contribution of these surface layers must be determined with an accuracy below 10% in order to be able to meet the targeted uncertainty budget for the redefinition. In this work, we performed a quantification of these surface layers, applying reference-free x-ray fluorescence spectrometry using a modified manipulator within an ultra-high vacuum chamber. Using this setup, we are able to quantitatively assess the surface of the spheres on a large area (about 20%) with quantification uncertainties below the required target of 10% for oxygen and carbon in order to meet an absolute uncertainty value of the mass below 10 µg.

  10. Highly c-axis-oriented monocrystalline Pb(Zr, Ti)O₃ thin films on si wafer prepared by fast cooling immediately after sputter deposition.

    PubMed

    Yoshida, Shinya; Hanzawa, Hiroaki; Wasa, Kiyotaka; Esashi, Masayoshi; Tanaka, Shuji

    2014-09-01

    We successfully developed sputter deposition technology to obtain a highly c-axis-oriented monocrystalline Pb(Zr, Ti)O3 (PZT) thin film on a Si wafer by fast cooling (~-180°C/min) of the substrate after deposition. The c-axis orientation ratio of a fast-cooled film was about 90%, whereas that of a slow-cooled (~-40°C/min) film was only 10%. The c-axis-oriented monocrystalline Pb(Zr0.5, Ti0.5)O3 films showed reasonably large piezoelectric coefficients, e(31,f) = ~-11 C/m(2), with remarkably small dielectric constants, ϵ(r) = ~220. As a result, an excellent figure of merit (FOM) was obtained for piezoelectric microelectromechanical systems (MEMS) such as a piezoelectric gyroscope. This c-axis orientation technology on Si will extend industrial applications of PZT-based thin films and contribute further to the development of piezoelectric MEMS.

  11. In-plane thermal conductivity of sub-20 nm thick suspended mono-crystalline Si layers.

    PubMed

    Ferrando-Villalba, P; Lopeandia, A F; Abad, Ll; Llobet, J; Molina-Ruiz, M; Garcia, G; Gerbolès, M; Alvarez, F X; Goñi, A R; Muñoz-Pascual, F J; Rodríguez-Viejo, J

    2014-05-09

    We measure the thermal conductivity of a 17.5-nm-thick single crystalline Si layer by using a suspended structure developed from a silicon-on-insulator wafer, in which the Si layer bridges the suspended platforms. The obtained value of 19 Wm(-1) K(-1) at room temperature represents a tenfold reduction with respect to bulk Si. This design paves the way for subsequent lateral nanostructuration of the layer with lithographic techniques, to define different geometries such as Si nanowires, nanostrips or phononic grids. As a proof of concept, nanostrips of 0.5 × 10 μm have been defined by focused ion beam (FIB) in the ultrathin Si layer. After the FIB cutting process with Ga ions at 30 kV and 100 pA, the measured thermal conductivity dramatically decreased to 1.7 Wm(-1) K(-1), indicating that the structure became severely damaged (amorphous). Re-crystallization of the structure was promoted by laser annealing while monitoring the Raman spectra. The thermal conductivity of the layer increased again to a value of 9.5 Wm(-1) K(-1) at room temperature, below that of the single crystalline material due to phonon scattering at the grain boundaries.

  12. New monocrystalline Si{sub 1-x}Ge{sub x} solar cells

    SciTech Connect

    Losada, B.R.; Moehlecke, A.; Ruiz, J.M.; Luque, A.

    1995-08-01

    The development of solar cells on Si{sub 1-x}Ge{sub x} might be interesting because they might present more current photo-response than the silicon cells, based on the lower bandgap of the alloyed crystal. In particular the use of Si{sub 1-x}Ge{sub x} solar cells in dual bandgap concentration structures as GaAs/Si{sub 1-x}Ge{sub x} can lead to total efficiency increase of about 1% as compared to the GaAs/Si structure, according to our calculations. Our effort is devoted to solar cells with low content of Ge, lower than 20% at. This choice is based on two previous hypothesis (1) A low content of Ge suggests that the well known silicon cell process, slightly modified, can be applied to the Si{sub 1-x}Ge{sub x} cells. (2) Calculations suggest that for utilisation in tandem with GaAs cells, the gain of efficiency is low above 20at % Ge.

  13. Experimental and computational investigation of microcrack behavior under combined environments in monocrystalline Si

    NASA Astrophysics Data System (ADS)

    Huang, W.-J.; Bringuier, S.; Paul, J.; Simmons-Potter, K.; Muralidharan, K.; Potter, B. G.

    2015-09-01

    An investigation of microindenter-induced crack evolution with independent variation of both temperature and relative humidity has been pursued in PV-grade Si wafers. Under static tensile strain conditions, an increase in subcritical crack elongation with increasing atmospheric water content was observed. To provide further insight into the potential physical and chemical conditions at the microcrack tip, micro-Raman measurements were performed. Preliminary results confirm a spatial variation in the frequency of the primary Si vibrational resonance within the cracktip region, associated with local stress state, whose magnitude is influenced by environmental conditions during the period of applied static strain. The experimental effort was paired with molecular dynamics (MD) investigations of microcrack evolution in single-crystal Si to furnish additional insight into mechanical contributions to crack elongation. The MD results demonstrate that crack-tip energetics and associated crack elongation velocity and morphology are intimately related to the crack and applied strain orientations with respect to the principal crystallographic axes. The resulting elastic strain energy release rate and the stress-strain response of the Si under these conditions form the basis for preliminary micro-scale peridynamics (PD) simulations of microcrack development under constant applied strain. These efforts will be integrated with the experimental results to further inform the mechanisms contributing to this important degradation mode in Si-based photovoltaics.

  14. In-plane thermal conductivity of sub-20 nm thick suspended mono-crystalline Si layers

    NASA Astrophysics Data System (ADS)

    Ferrando-Villalba, P.; Lopeandia, A. F.; Abad, Ll; Llobet, J.; Molina-Ruiz, M.; Garcia, G.; Gerbolès, M.; Alvarez, F. X.; Goñi, A. R.; Muñoz-Pascual, F. J.; Rodríguez-Viejo, J.

    2014-05-01

    We measure the thermal conductivity of a 17.5-nm-thick single crystalline Si layer by using a suspended structure developed from a silicon-on-insulator wafer, in which the Si layer bridges the suspended platforms. The obtained value of 19 Wm-1 K-1 at room temperature represents a tenfold reduction with respect to bulk Si. This design paves the way for subsequent lateral nanostructuration of the layer with lithographic techniques, to define different geometries such as Si nanowires, nanostrips or phononic grids. As a proof of concept, nanostrips of 0.5 × 10 μm have been defined by focused ion beam (FIB) in the ultrathin Si layer. After the FIB cutting process with Ga ions at 30 kV and 100 pA, the measured thermal conductivity dramatically decreased to 1.7 Wm-1 K-1, indicating that the structure became severely damaged (amorphous). Re-crystallization of the structure was promoted by laser annealing while monitoring the Raman spectra. The thermal conductivity of the layer increased again to a value of 9.5 Wm-1 K-1 at room temperature, below that of the single crystalline material due to phonon scattering at the grain boundaries.

  15. Improvement of minority carrier life time in N-type monocrystalline Si by the Czochralski method

    NASA Astrophysics Data System (ADS)

    Baik, Sungsun; Pang, Ilsun; Kim, Jaemin; Kim, Kwanghun

    2016-07-01

    The installation amount of solar power plants increases every year. Multi-crystalline Si solar cells comprise a large share of the market of solar power plants. Multi-crystalline and single-crystalline Si solar cells are competing against one another in the market. Many single-crystalline companies are trying to develop and produce n-type solar cells with higher cell efficiency than that of p-type. In n-type wafers with high cell efficiency, wafer quality has become increasingly important. In order to make ingots with higher MCLT, the effects of both poly types related to metal impurities and pull speeds related to vacancy concentration on minority carrier life time were studied. In the final part of ingots, poly types related to the metal impurities are a dominant factor on MCLT. In the initial part of ingots, pull speeds related to vacancy concentration are a dominant factor on MCLT. [Figure not available: see fulltext.

  16. Design of monocrystalline Si/SiGe multi-quantum well microbolometer detector for infrared imaging systems

    NASA Astrophysics Data System (ADS)

    Shafique, Atia; Durmaz, Emre C.; Cetindogan, Barbaros; Yazici, Melik; Kaynak, Mehmet; Kaynak, Canan B.; Gurbuz, Yasar

    2016-05-01

    This paper presents the design, modelling and simulation results of silicon/silicon-germanium (Si/SiGe) multi-quantum well based bolometer detector for uncooled infrared imaging system. The microbolometer is designed to detect light in the long wave length infrared (LWIR) range from 8 to 14 μm with pixel size of 25 x 25 μm. The design optimization strategy leads to achieve the temperature coefficient of resistance (TCR) 4.5%/K with maximum germanium (Ge) concentration of 50%. The design of microbolometer entirely relies on standard CMOS and MEMS processes which makes it suitable candidate for commercial infrared imaging systems.

  17. Buried Porous Silicon-Germanium Layers in Monocrystalline Silicon Lattices

    NASA Technical Reports Server (NTRS)

    Fathauer, Robert W. (Inventor); George, Thomas (Inventor); Jones, Eric W. (Inventor)

    1998-01-01

    Monocrystalline semiconductor lattices with a buried porous semiconductor layer having different chemical composition is discussed and monocrystalline semiconductor superlattices with a buried porous semiconductor layers having different chemical composition than that of its monocrystalline semiconductor superlattice are discussed. Lattices of alternating layers of monocrystalline silicon and porous silicon-germanium have been produced. These single crystal lattices have been fabricated by epitaxial growth of Si and Si-Ge layers followed by patterning into mesa structures. The mesa structures are strain etched resulting in porosification of the Si-Ge layers with a minor amount of porosification of the monocrystalline Si layers. Thicker Si-Ge layers produced in a similar manner emitted visible light at room temperature.

  18. Photocarrier radiometry for predicting the degradation of electrical parameters of monocrystalline silicon (c-Si) solar cell irradiated by 100 KeV proton beams

    NASA Astrophysics Data System (ADS)

    Song, P.; Liu, J. Y.; Yuan, H. M.; Oliullah, Md.; Wang, F.; Wang, Y.

    2016-09-01

    In this study, the monocrystalline silicon (c-Si) solar cell irradiated by 100 KeV proton beams at various fluences is investigated. A one-dimensional two-layer carrier density wave model has been developed to estimate the minority carrier lifetime of n-region and p-region of the non-irradiated c-Si solar cell by best fitting with the experimental photocarrier radiometry (PCR) signal (the amplitude and the phase). Furthermore, the lifetime is used to determine the initial defect density of the quasi-neutral region (QNR) of the solar cell to predict its I-V characteristics. The theoretically predicted short-circuit current density (Jsc), and open-circuit voltage (Voc) of the non-irradiated samples are in good agreement with experiment. Then a three-region defect distribution model for the c-Si solar cell irradiated by proton beams is carried out to describe the defect density distribution according to Monte Carlo simulation results and the initial defect density of the non-irradiated sample. Finally, we find that the electrical measurements of Jsc and Voc of the solar cells irradiated at different fluences using 100 KeV proton beams are consistent with the PCR predicting results.

  19. Characteristics of poly- and mono-crystalline BeO and SiO2 as thermal and cold neutron filters

    NASA Astrophysics Data System (ADS)

    Adib, M.; Habib, N.; Bashter, I. I.; Morcos, H. N.; El-Mesiry, M. S.; Mansy, M. S.

    2015-09-01

    A simple model along with a computer code "HEXA-FILTERS" is used to carry out the calculation of the total cross-sections of BeO and SiO2 having poly or mono-crystalline form as a function of neutron wavelength at room (R.T.) and liquid nitrogen (L.N.) temperatures. An overall agreement is indicated between the calculated neutron cross-sections and experimental data. Calculation shows that 25 cm thick of polycrystalline BeO cooled at liquid nitrogen temperature was found to be a good filter for neutron wavelengths longer than 0.46 nm. While, 50 cm of SiO2, with much less transmission, for neutrons with wavelengths longer than 0.85 nm. It was also found that 10 cm of BeO and 15 cm SiO2 thick mono-crystals cut along their (0 0 2) plane, with 0.5° FWHM on mosaic spread and cooled at L.N., are a good thermal neutron filter, with high effect-to-noise ratio.

  20. Effect of cutting temperature on hardness of SiC and diamond in the nano-cutting process of monocrystalline silicon

    NASA Astrophysics Data System (ADS)

    Wang, Jiachun; Li, Yuntao; Liu, Xiaoxuan; Lv, Maoqiang

    2016-10-01

    In the process of cutting silicon by natural diamond tools, groove wear happens on the flank face of cutting tool frequently.Scholars believe that one of the wear reasons is mechanical scratching effect by hard particles like SiC. To reveal the mechanical scratching mechanism, it is essential to study changes in the mechanical properties of hard particles and diamond, especially the effect of cutting temperature on hardness of diamond and hard particles. Molecular dynamics (MD) model that contact-zone temperature between tool and workpiece was calculated by dividing zone while nano-cutting monocrystalline silicon was established, cutting temperature values in different regions were computed as the simulation was carried out.On this basis, the models of molecular dynamics simulation of SiC and diamond were established separately with setting the initial temperature to room temperature. The laws of length change of C-C bond and Si-C bond varing with increase of simulation temperature were studied. And drawing on predecessors' research on theoretical calculation of hardness of covalent crystals and the relationship between crystal valence electron density and bond length, the curves that the hardness of diamond and SiC varing with bond length were obtained. The effect of temperature on the hardness was calculated. Results show that, local cutting temperature can reach 1300K.The rise in cutting temperature leaded to a decrease in the diamond local atomic clusters hardness,SiC local atomic clusters hardness increased. As the cutting temperature was more than 1100K,diamond began to soften, the local clusters hardness was less than that of SiC.

  1. Photocarrier Radiometry for Noncontact Evaluation of Monocrystalline Silicon (c-Si) Solar Cell Irradiated by 1 MeV Electron Beams

    NASA Astrophysics Data System (ADS)

    Song, P.; Liu, J. Y.; Yuan, H. M.; Wang, F.; Wang, Y.

    2016-08-01

    In this paper, the monocrystalline silicon (c-Si) solar cell irradiated by 1 MeV electron beams was investigated using noncontact photocarrier radiometry (PCR). A theoretical 1D two-layer PCR model including the impedance effect of the p-n junction was used to characterize the transport properties (carrier lifetime, diffusion coefficient, and surface recombination velocities) of c-Si solar cells irradiated by 1 MeV electron beams with different fluences. The carrier transport parameters were derived by the best fit through PCR measurements. Furthermore, an Ev+0.56 eV trap was introduced into the band gap based on the minority carrier lifetime reduction. An I-V characteristic was obtained by both AFORS-HET simulation and experimental study, and the simulation results shows in good agreement with the experimental results. Moreover, the simulation and experiment results also indicate that the increase of fluences of electron beams results in the reduction of short-circuit current and open-circuit voltage.

  2. Growth, Characterization and Device Development in Monocrystalline Diamond Films

    DTIC Science & Technology

    1992-09-01

    AD-A256 283 Quarterly Letter Report Growth, Characterization and Device Development in Monocrystalline Diamond Films DT C * F LEC EGOT 2 1992L... Characterization and Device Development in s400003srrO8 Monocrystalline Diamond Films 1114SS 6. AUTHOR(S) N00179N66005 Robert F. Davis 4B855 7. PERFORMING...deposited on single crystal Si(100) substrates via in-situ carburization followed by bias-enhanced nucleation. Photoluminescence and Raman spectroscopy

  3. The Seebeck coefficient of monocrystalline α-SiC and polycrystalline β-SiC measured at 300-533 K

    NASA Astrophysics Data System (ADS)

    Abu-Ageel, N.; Aslam, M.; Ager, R.; Rimai, L.

    2000-01-01

    The temperature dependence of the Seebeck coefficient of polycrystalline icons/Journals/Common/beta" ALT="beta" ALIGN="TOP"/> -SiC films deposited on quartz substrates by laser ablation and of commercially available icons/Journals/Common/alpha" ALT="alpha" ALIGN="TOP"/> -SiC wafers is reported in a temperature range of 300-533 K for the first time. The Seebeck emf of icons/Journals/Common/alpha" ALT="alpha" ALIGN="TOP"/> -SiC substrates and icons/Journals/Common/beta" ALT="beta" ALIGN="TOP"/> -SiC samples ranges between -9 µV °C-1 and -108 µV °C-1 which is higher than that of commercial Pt thermocouples.

  4. Method of producing buried porous silicon-geramanium layers in monocrystalline silicon lattices

    NASA Technical Reports Server (NTRS)

    Fathauer, Robert W. (Inventor); George, Thomas (Inventor); Jones, Eric W. (Inventor)

    1997-01-01

    Lattices of alternating layers of monocrystalline silicon and porous silicon-germanium have been produced. These single crystal lattices have been fabricated by epitaxial growth of Si and Si--Ge layers followed by patterning into mesa structures. The mesa structures are stain etched resulting in porosification of the Si--Ge layers with a minor amount of porosification of the monocrystalline Si layers. Thicker Si--Ge layers produced in a similar manner emitted visible light at room temperature.

  5. Quasi-monocrystalline silicon for thin-film devices

    NASA Astrophysics Data System (ADS)

    Rinke, T. J.; Bergmann, R. B.; Werner, J. H.

    Thermal crystallization of a double layer porous Si film creates a monocrystalline Si film with a thin separation layer between the Si film and the reusable starting wafer. The process enables transfer of thin monocrystalline Si films to foreign substrates, whereby devices may be formed before or after separation of the film. Sub-micrometer thick films are almost compact, while films with a thickness of several μm contain voids, and are therefore termed ``quasi-monocrystalline''. Internal voids strongly enhance optical absorption by light scattering. The hole mobility is 78 cm2V-1s-1 at a p-type starting wafer resistivity of 0.05 Ωcm.

  6. Monocrystalline germanium film on sapphire

    NASA Astrophysics Data System (ADS)

    Godbey, David J.; Qadri, Syed B.

    1993-04-01

    A monocrystalline germanium film is grown on a sapphire substrate with a (I 102) orientation. The substrate is first pretreated to restructure the (1102) surface plane. Typically, restructuring is accomplished by either an anneal at high temperature or ion bombardment. A monocrystalline germanium layer is grown on the pretreated surface by a vapor deposition process such as molecular beam epitaxy or chemical vapor deposition.

  7. Methods for manufacturing monocrystalline or near-monocrystalline cast materials

    DOEpatents

    Stoddard, Nathan G

    2014-04-29

    Methods are provided for casting one or more of a semiconductor, an oxide, and an intermetallic material. With such methods, a cast body of a monocrystalline form of the one or more of a semiconductor, an oxide, and an intermetallic material may be formed that is free of, or substantially free of, radially-distributed impurities and defects and having at least two dimensions that are each at least about 35 cm.

  8. Development of brazing foils to join monocrystalline tungsten alloys with ODS-EUROFER steel

    NASA Astrophysics Data System (ADS)

    Kalin, B. A.; Fedotov, V. T.; Sevrjukov, O. N.; Kalashnikov, A. N.; Suchkov, A. N.; Moeslang, A.; Rohde, M.

    2007-08-01

    Results on rapidly solidified filler metals for brazing W with W and monocrystalline W with EUROFER steel (FS) are presented. Rapidly quenched powder-type filler metals based on Ti bal-V-Cr-Be were developed to braze polycrystalline W with monocrystalline W. In addition, Fe bal-Ta-Ge-Si-B-Pd alloys were developed to braze monocrystalline W with FS for helium gas cooled divertors and plasma-facing components. The W to FS brazed joints were fabricated under vacuum at 1150 °C, using a Ta spacer of 0.1 mm in thickness to account for the different thermal expansions. The monocrystalline tungsten as well as the related brazed joints withstood 30 cycles between 750 °C/20 min and air cooling/3-5 min.

  9. Laser compression of monocrystalline tantalum

    NASA Astrophysics Data System (ADS)

    Lu, Chia-Hui; Remington, B.; Maddox, Brian; Kad, Bimal; Park, Hye-Sook; Prisbrey, S. T.; Luo, Rain; Meyers, Marc

    2012-03-01

    Monocrystalline tantalum with orientations [100] and [111] was subjected to laser driven compression at laser energies of 350 to 685 J, generating shock amplitudes varying from 15 to 100 GPa. The laser beam, with a beam spot diameter of ~1 mm, created a crater of significant depth (~ 80 to ~ 200 μm). Twins were observed just below the crater surface (~ 42 μm) by back-scattered SEM. Transmission electron microscopy (TEM) revealed profuse mechanical twinning within a distance from the energy deposition surface of ~ 1.5 mm at 684 J compression power, corresponding to an approximate pressure of 35 GPa. The decay of the pulse through the specimens was accompanied by an attendant decrease in the density of shock-generated dislocations. Microhardness measurements were conducted on the recovered samples. The experimentally measured dislocation densities and threshold stress for twinning are compared with predictions using analyses based on the constitutive response and the similarities and differences are discussed in terms of the mechanisms of defect generation.

  10. Laser Compression of Monocrystalline Tantalum

    NASA Astrophysics Data System (ADS)

    Meyers, Marc; Lu, Chia-Hui; Remington, Bruce; Maddox, Brian; Park, Hye-Sook; Priesbrey, Shon; Kad, Bimal; Luo, Rain

    2011-06-01

    Monocrystalline tantalum with orientations [100] and [111] was subjected to laser driven compression at laser energies of 350 to 685 J, generating shock amplitudes varying from 15 to 100 GPa. The laser beam, with a beam spot diameter of ~1 mm, created a crater of significant depth (~80 to ~200 μm). Twins were observed just below the crater surface (~ 42 μm) by back-scattered SEM. Transmission electron microscopy (TEM) revealed profuse mechanical twinning within a distance from the energy deposition surface of about 1.5 mm (~1.3 mm from residual crater vertex) at 684 J compression power, corresponding to an approximate pressure of 35 GPa. The decay of the pulse through the specimens was accompanied by an attendant decrease in the density of shock-generated dislocations. Dislocation densities as a function of pressure were calculated for the case of homogeneous nucleation and for Orowan hardening. The observed results are compared with predictions. Microhardness measurements were conducted on the recovered samples. The experimentally measured threshold stress for twinning is compared with predictions using an analysis based on the constitutive response and the similarities and differences are discussed.

  11. Targeting siRNA Missiles to HER2+ Breast Cancer

    DTIC Science & Technology

    2007-06-01

    Breast Cancer PRINCIPAL INVESTIGATOR: Lali K. Medina-Kauwe, Ph.D. CONTRACTING ORGANIZATION: Cedars-Sinai Medical Center...5a. CONTRACT NUMBER Targeting siRNA Missiles to HER2+ Breast Cancer 5b. GRANT NUMBER W81XWH-06-1-0549 5c. PROGRAM ELEMENT NUMBER 6...that delivery conjugates can be assembled that can direct siRNA molecules to target cells, including HER2+ human breast cancer cells, in culture in

  12. Nanotechnology for in vivo targeted siRNA delivery.

    PubMed

    Dahlman, James E; Kauffman, Kevin J; Langer, Robert; Anderson, Daniel G

    2014-01-01

    Small interfering RNAs (siRNAs) can specifically inhibit gene expression. As a result, they have tremendous scientific and clinical potential. However, the use of these molecules in patients and animal models has been limited by challenges with delivery. Intracellular RNA delivery is difficult; it requires a system that protects the siRNA from degradative nucleases in the bloodstream, minimizes clearance by the reticuloendothelial system, maximizes delivery to the target tissue, and promotes entry into, and out of, an endocytic vesicle. Despite these barriers, recent data suggest that RNA may be targeted to cells of interest in vivo. Herein we outline strategies for targeted siRNA delivery, and describe how these strategies may be improved.

  13. The SiC Direct Target Prototype for SPES

    SciTech Connect

    Rizzi, V.; Andrighetto, A.; Barbui, M.; Carturan, S.; Cinausero, M.; Giacchini, M.; Gramegna, F.; Lollo, M.; Maggioni, G.; Prete, G.; Tonezzer, M.; Antonucci, C.; Cevolani, S.; Petrovich, C.; Biasetto, L.; Colombo, P.; Manzolaro, M.; Meneghetti, M.; Celona, L.; Chines, F.

    2007-10-26

    A R and D study for the realization of a Direct Target is in progress within the SPES project for RIBs production at the Laboratori Nazionali of Legnaro. A proton beam (40 MeV energy, 0.2 mA current) is supposed to impinge directly on a UCx multiple thin disks target, the power released by the proton beam is dissipated mainly through irradiation. A SiC target prototype with a 1:5 scale has been developed and tested. Thermal, mechanical and release calculations have been performed to fully characterize the prototype. An online test has been performed at the HRIBF facility of the Oak Ridge National Laboratory (ORNL), showing that our SiC target can sustain a proton beam current considerably higher than the maximum beam current used with the standard HRIBF target configuration.

  14. The SiC Direct Target Prototype for SPES

    SciTech Connect

    Rizzi, Valentina; Andrighetto, Alberto; Antonucci, C.; Barbui, Marina; Biasetto, Lisa; Carturan, S.; Celona, L.; Cevolani, S.; Chines, Francesco; Cinausero, Marco; Colombo, P.; Cuttone, G.; Di Bernardo, P.; Giacchini, Mauro; Gramegna, Fabiana; Lollo, M.; Maggioni, G.; Manzolaro, Mattia; Meneghetti, G.; Messina, G. Esteban; Petrovich, C.; Piga, L.; Prete, Gianfranco; Re, Maurizio; Rizzo, D.; Stracener, Daniel W; Tonezzer, Michele; Zanonato, P.

    2007-01-01

    A R&D study for the realization of a Direct Target is in progress within the SPES project for RIBs production at the Laboratori Nazionali of Legnaro. A proton beam (40 MeV energy, 0.2 niA current) is supposed to impinge directly on a UCx multiple thin disks target, the power released by the proton beam is dissipated mainly through irradiation. A SiC target prototype with a 1:5 scale has been developed and tested. Thermal, mechanical and release calculations have been performed to fully characterize the prototype. An online test has been performed at the HRIBF facility of the Oak Ridge National Laboratory (ORNL), showing that our Sic target can sustain a proton beam current considerably higher than the maximum beam current used with the standard HRIBF target configuration.

  15. Enhancing potency of siRNA targeting fusion genes by optimization outside of target sequence

    PubMed Central

    Gavrilov, Kseniya; Seo, Young-Eun; Tietjen, Gregory T.; Cui, Jiajia; Cheng, Christopher J.; Saltzman, W. Mark

    2015-01-01

    Canonical siRNA design algorithms have become remarkably effective at predicting favorable binding regions within a target mRNA, but in some cases (e.g., a fusion junction site) region choice is restricted. In these instances, alternative approaches are necessary to obtain a highly potent silencing molecule. Here we focus on strategies for rational optimization of two siRNAs that target the junction sites of fusion oncogenes BCR-ABL and TMPRSS2-ERG. We demonstrate that modifying the termini of these siRNAs with a terminal G-U wobble pair or a carefully selected pair of terminal asymmetry-enhancing mismatches can result in an increase in potency at low doses. Importantly, we observed that improvements in silencing at the mRNA level do not necessarily translate to reductions in protein level and/or cell death. Decline in protein level is also heavily influenced by targeted protein half-life, and delivery vehicle toxicity can confound measures of cell death due to silencing. Therefore, for BCR-ABL, which has a long protein half-life that is difficult to overcome using siRNA, we also developed a nontoxic transfection vector: poly(lactic-coglycolic acid) nanoparticles that release siRNA over many days. We show that this system can achieve effective killing of leukemic cells. These findings provide insights into the implications of siRNA sequence for potency and suggest strategies for the design of more effective therapeutic siRNA molecules. Furthermore, this work points to the importance of integrating studies of siRNA design and delivery, while heeding and addressing potential limitations such as restricted targetable mRNA regions, long protein half-lives, and nonspecific toxicities. PMID:26627251

  16. Methods and apparatuses for manufacturing monocrystalline cast silicon and monocrystalline cast silicon bodies for photovoltaics

    DOEpatents

    Stoddard, Nathan G [Gettysburg, PA

    2011-11-01

    Methods and apparatuses are provided for casting silicon for photovoltaic cells and other applications. With such methods and apparatuses, a cast body of monocrystalline silicon may be formed that is free of, or substantially free of, radially-distributed impurities and defects and having at least two dimensions that are each at least about 35 cm is provided.

  17. Methods and apparatus for manufacturing monocrystalline cast silicon and monocrystalline cast silicon bodies for photovoltaics

    DOEpatents

    Stoddard, Nathan G

    2014-01-14

    Methods and apparatuses are provided for casting silicon for photovoltaic cells and other applications. With such methods and apparatuses, a cast body of monocrystalline silicon may be formed that is free of, or substantially free of, radially-distributed impurities and defects and having at least two dimensions that are each at least about 35 cm is provided.

  18. Targeting siRNA Missiles to Her2+ Breast Cancer

    DTIC Science & Technology

    2009-06-01

    testing for targeted toxicity of HerPBK10-siRNA, we assessed cytotoxic potential in vivo after intratumoral (IT) injection of complexes in tumor-bearing...single tail vein injection of Dox alone or the complex (HerDox) (0.02 mg/kg with respect to Dox conc) and were imaged live using a customized macro...mice were injected 0.02 mg/kg (final Dox dose) of HerDox ox via the tail vein and imaged with a om small animal imager. A, Imaging of mice after

  19. Molecular dynamics investigations of mechanical behaviours in monocrystalline silicon due to nanoindentation at cryogenic temperatures and room temperature.

    PubMed

    Du, Xiancheng; Zhao, Hongwei; Zhang, Lin; Yang, Yihan; Xu, Hailong; Fu, Haishuang; Li, Lijia

    2015-11-05

    Molecular dynamics simulations of nanoindentation tests on monocrystalline silicon (010) surface were conducted to investigate the mechanical properties and deformation mechanism from cryogenic temperature being 10 K to room temperature being 300 K. Furthermore, the load-displacement curves were obtained and the phase transformation was investigated at different temperatures. The results show that the phase transformation occurs both at cryogenic temperatures and at room temperature. By searching for the presence of the unique non-bonded fifth neighbour atom, the metastable phases (Si-III and Si-XII) with fourfold coordination could be distinguished from Si-I phase during the loading stage of nanoindentation process. The Si-II, Si-XIII, and amorphous phase were also found in the region beneath the indenter. Moreover, through the degree of alignment of the metastable phases along specific crystal orientation at different temperatures, it was found that the temperature had effect on the anisotropy of the monocrystalline silicon, and the simulation results indicate that the anisotropy of monocrystalline silicon is strengthened at low temperatures.

  20. Molecular dynamics investigations of mechanical behaviours in monocrystalline silicon due to nanoindentation at cryogenic temperatures and room temperature

    PubMed Central

    Du, Xiancheng; Zhao, Hongwei; Zhang, Lin; Yang, Yihan; Xu, Hailong; Fu, Haishuang; Li, Lijia

    2015-01-01

    Molecular dynamics simulations of nanoindentation tests on monocrystalline silicon (010) surface were conducted to investigate the mechanical properties and deformation mechanism from cryogenic temperature being 10 K to room temperature being 300 K. Furthermore, the load-displacement curves were obtained and the phase transformation was investigated at different temperatures. The results show that the phase transformation occurs both at cryogenic temperatures and at room temperature. By searching for the presence of the unique non-bonded fifth neighbour atom, the metastable phases (Si-III and Si-XII) with fourfold coordination could be distinguished from Si-I phase during the loading stage of nanoindentation process. The Si-II, Si-XIII, and amorphous phase were also found in the region beneath the indenter. Moreover, through the degree of alignment of the metastable phases along specific crystal orientation at different temperatures, it was found that the temperature had effect on the anisotropy of the monocrystalline silicon, and the simulation results indicate that the anisotropy of monocrystalline silicon is strengthened at low temperatures. PMID:26537978

  1. siRNA-based topical microbicides targeting sexually transmitted diseases

    PubMed Central

    Katakowski, Joseph A

    2012-01-01

    Sexually transmitted infections (STIs) are a major cause of morbidity and mortality worldwide. Although a vaccine is available for HPV, no effective vaccines exist for the HIV-1 and HSV-2 viral pathogens, and there are no cures for these infections. Furthermore, recent setbacks in clinical trials, such as the failure of the STEP trial to prevent HIV-1 infection, have emphasized the need to develop alternative approaches to interrupt transmission of these pathogens. One alternative strategy is represented by the use of topically applied microbicides, and such agents are being developed against various viruses. RNAi-based microbicides have recently been demonstrated to prevent HSV-2 transmission, and may be useful for targeting multiple STIs. In this review, microbicides that are under development for the prevention of STIs are described, with a focus on topically applied microbicidal siRNAs. PMID:20373263

  2. Damage Mechanics Model Development for Monocrystalline Superalloys (PREPRINT)

    DTIC Science & Technology

    2010-02-01

    AFRL-RX-WP-TP-2010-4148 DAMAGE MECHANICS MODEL DEVELOPMENT FOR MONOCRYSTALLINE SUPERALLOYS (PREPRINT) Mark A. Tschopp Mississippi State...October 2009 4. TITLE AND SUBTITLE DAMAGE MECHANICS MODEL DEVELOPMENT FOR MONOCRYSTALLINE SUPERALLOYS (PREPRINT) 5a. CONTRACT NUMBER FA8650-07-D-5800...better predict damage initiation, such as cracking, in superalloys under engine representative conditions. This report details work that focuses on

  3. Targeted tumor-penetrating siRNA nanocomplexes for credentialing the ovarian cancer target ID4

    PubMed Central

    Ren, Yin; Cheung, Hiu Wing; von Maltzhan, Geoffrey; Agrawal, Amit; Cowley, Glenn S.; Weir, Barbara A.; Boehm, Jesse S.; Tamayo, Pablo; Karst, Alison M.; Liu, Joyce F.; Hirsch, Michelle S.; Mesirov, Jill P.; Drapkin, Ronny; Root, David E.; Lo, Justin; Fogal, Valentina; Ruoslahti, Erkki; Hahn, William C.; Bhatia, Sangeeta N.

    2013-01-01

    The comprehensive characterization of a large number of cancer genomes will eventually lead to a compendium of genetic alterations in specific cancers. Unfortunately, the number and complexity of identified alterations complicate endeavors to identify biologically relevant mutations critical for tumor maintenance, because many of these targets are not amenable to manipulation by small molecules or antibodies. RNAi provides a direct way to study putative cancer targets; however, specific delivery of therapeutics to the tumor parenchyma remains an intractable problem. We describe a platform for the discovery and initial validation of cancer targets, composed of a systematic effort to identify amplified and essential genes in human cancer cell lines and tumors partnered with a novel modular delivery technology. We developed a tumor-penetrating nanocomplex (TPN) comprised of siRNA complexed with a tandem tumor-penetrating and membrane-translocating peptide, which enabled the specific delivery of siRNA deep into the tumor parenchyma. We employed TPN in vivo to evaluate inhibitor of DNA binding 4 (ID4) as a novel oncogene. Treatment of ovarian tumor-bearing mice with ID4-specific TPN suppressed growth of established tumors and significantly improved survival. These observations not only credential ID4 as an oncogene in 32% of high-grade ovarian cancers, but also provide a framework for the identification, validation, and understanding of potential therapeutic cancer targets. PMID:22896676

  4. Multi-target siRNA: Therapeutic Strategy for Hepatocellular Carcinoma

    PubMed Central

    Li, Tiejun; Xue, Yuwen; Wang, Guilan; Gu, Tingting; Li, Yunlong; Zhu, York Yuanyuan; Chen, Li

    2016-01-01

    Multiple targets RNAi strategy is a preferred way to treat multigenic diseases, especially cancers. In the study, multi-target siRNAs were designed to inhibit NET-1, EMS1 and VEGF genes in hepatocellular carcinoma (HCC) cells. And multi-target siRNAs showed better silencing effects on NET-1, EMS1 and VEGF, compared with single target siRNA. Moreover, multi-target siRNA showed greater suppression effects on proliferation, migration, invasion, angiogenesis and induced apoptosis in HCC cells. The results suggested that multi-target siRNA might be a preferred strategy for cancer therapy and NET-1, EMS1 and VEGF could be effective targets for HCC treatments. PMID:27390607

  5. Minimizing off-target effects by using diced siRNAs for RNA interference

    PubMed Central

    Myers, Jason W; Chi, Jen-Tsan; Gong, Delquin; Schaner, Marci E; Brown, Patrick O; Ferrell, James E

    2006-01-01

    Microarray studies have shown that individual synthetic small interfering RNAs (siRNAs) can have substantial off-target effects. Pools of siRNAs, produced by incubation of dsRNAs with recombinant Dicer or RNase III, can also be used to silence genes. Here we show that diced siRNA pools are highly complex, containing hundreds of different individual siRNAs. This high complexity could either compound the problem of off-target effects, since the number of potentially problematic siRNAs is high, or it could diminish the problem, since the concentration of any individual problematic siRNA is low. We therefore compared the off-target effects of diced siRNAs to chemically synthesized siRNAs. In agreement with previous reports, we found that two chemically synthesized siRNAs targeted against p38α MAPK (MAPK14) induced off-target changes in the abundance of hundreds of mRNAs. In contrast, three diced siRNA pools against p38α MAPK had almost no off-target effects. The off-target effects of a synthetic siRNA were reduced when the siRNA was diluted 3-fold in a diced pool and completely alleviated when it was diluted 30- or 300-fold, suggesting that when problematic siRNAs are present within a diced pool, their absolute concentration is too low to result in significant off-target effects. These data rationalize the observed high specificity of RNA interference in C. elegans and D. melanogaster, where gene suppression is mediated by endogenously-generated diced siRNA pools, and provide a strategy for improving the specificity of RNA interference experiments and screens in mammalian cells. PMID:19771225

  6. Development of siRNA payloads to target KRAS-mutant cancer

    PubMed Central

    Ritchie, Cayde D.; Thapar, Vishal; Lee, Liam C.; Hsu, Dennis J.; Grace, Danielle; Carver, Joseph O.; Zuber, Johannes; Luo, Ji; McCormick, Frank; Lowe, Scott W.

    2014-01-01

    RNA interference (RNAi) is a powerful tool for target identification and can lead to novel therapies for pharmacologically intractable targets such as KRAS. RNAi therapy must combine potent siRNA payloads with reliable in vivo delivery for efficient target inhibition. We employed a functional “Sensor” assay to establish a library of potent siRNAs against RAS pathway genes and show they efficiently suppress their targets at low dose. This reduces off-target effects and enables combination gene knockdown. We administered Sensor siRNAs in vitro and in vivo and validated the delivery of KRAS siRNA alone and siRNA targeting the complete RAF effector node (A/B/C-RAF) as promising strategies to treat KRAS-mutant colorectal cancer. We further demonstrate that improved therapeutic efficacy is achieved by formulating siRNA payloads that combine both single-gene siRNA and node-targeted siRNAs (KRAS+PIK3C-A/B). The customizable nature of Sensor siRNA payloads offers a universal platform for combination target identification and development of RNAi therapeutics. PMID:25100204

  7. Magnetic properties of Fe/FeSi2/Fe3Si trilayered films prepared by facing targets sputtering deposition

    NASA Astrophysics Data System (ADS)

    Ishibashi, Kazuya; Nakashima, Kazutoshi; Sakai, Ken-Ichiro; Yoshitake, Tsuyoshi

    2015-09-01

    Whereas giant magnetoresistance and tunnel magnetoresistance films generally employ nonmagnetic metal and insulator spacers, respectively, we have studied Fe3Si/FeSi artificial lattices, in which FeSi2 is semiconducting and its employment as spacers is specific to our research. For the formation of parallel/antiparallel alignments of layer magnetizations, the employment of ferromagnetic layers with different coercive forces is required. There have been few studies on the fabrication of Fe-Si system spin valves comprising ferromagnetic layers with different coercive forces. In this work, Fe3Si and Fe were employed as ferromagnetic layer materials with different coercive forces. Fe/FeSi2/Fe3Si trilayered spin valve junctions by facing targets direct-current sputtering deposition combined with a mask method, and their electrical and magnetic properties were studied. An Fe3Si layer was epitaxially grown on Si(111) substrate as a bottom layer. After that, An Fe layer with a large coercive force was deposited as a top layer, posterior to a FeSi2 layer being deposited. From magnetization curves measured by a vibrating sample magnetometer, it was confirmed that the parallel and antiparallel magnetization alignments of ferromagnetic layers are clearly realized. This work was supported by JSPS KAKENHI Grant Number 15K21594.

  8. The siRNA Non-seed Region and Its Target Sequences Are Auxiliary Determinants of Off-Target Effects

    PubMed Central

    Kamola, Piotr J.; Nakano, Yuko; Takahashi, Tomoko; Wilson, Paul A.; Ui-Tei, Kumiko

    2015-01-01

    RNA interference (RNAi) is a powerful tool for post-transcriptional gene silencing. However, the siRNA guide strand may bind unintended off-target transcripts via partial sequence complementarity by a mechanism closely mirroring micro RNA (miRNA) silencing. To better understand these off-target effects, we investigated the correlation between sequence features within various subsections of siRNA guide strands, and its corresponding target sequences, with off-target activities. Our results confirm previous reports that strength of base-pairing in the siRNA seed region is the primary factor determining the efficiency of off-target silencing. However, the degree of downregulation of off-target transcripts with shared seed sequence is not necessarily similar, suggesting that there are additional auxiliary factors that influence the silencing potential. Here, we demonstrate that both the melting temperature (Tm) in a subsection of siRNA non-seed region, and the GC contents of its corresponding target sequences, are negatively correlated with the efficiency of off-target effect. Analysis of experimentally validated miRNA targets demonstrated a similar trend, indicating a putative conserved mechanistic feature of seed region-dependent targeting mechanism. These observations may prove useful as parameters for off-target prediction algorithms and improve siRNA ‘specificity’ design rules. PMID:26657993

  9. Self-assembly of microscopic chiplets at a liquid-liquid-solid interface forming a flexible segmented monocrystalline solar cell.

    PubMed

    Knuesel, Robert J; Jacobs, Heiko O

    2010-01-19

    This paper introduces a method for self-assembling and electrically connecting small (20-60 micrometer) semiconductor chiplets at predetermined locations on flexible substrates with high speed (62500 chips/45 s), accuracy (0.9 micrometer, 0.14 degrees), and yield (> 98%). The process takes place at the triple interface between silicone oil, water, and a penetrating solder-patterned substrate. The assembly is driven by a stepwise reduction of interfacial free energy where chips are first collected and preoriented at an oil-water interface before they assemble on a solder-patterned substrate that is pulled through the interface. Patterned transfer occurs in a progressing linear front as the liquid layers recede. The process eliminates the dependency on gravity and sedimentation of prior methods, thereby extending the minimal chip size to the sub-100 micrometer scale. It provides a new route for the field of printable electronics to enable the integration of microscopic high performance inorganic semiconductors on foreign substrates with the freedom to choose target location, pitch, and integration density. As an example we demonstrate a fault-tolerant segmented flexible monocrystalline silicon solar cell, reducing the amount of Si that is used when compared to conventional rigid cells.

  10. Self-assembly of microscopic chiplets at a liquid–liquid–solid interface forming a flexible segmented monocrystalline solar cell

    PubMed Central

    Knuesel, Robert J.; Jacobs, Heiko O.

    2010-01-01

    This paper introduces a method for self-assembling and electrically connecting small (20–60 micrometer) semiconductor chiplets at predetermined locations on flexible substrates with high speed (62500 chips/45 s), accuracy (0.9 micrometer, 0.14°), and yield (> 98%). The process takes place at the triple interface between silicone oil, water, and a penetrating solder-patterned substrate. The assembly is driven by a stepwise reduction of interfacial free energy where chips are first collected and preoriented at an oil-water interface before they assemble on a solder-patterned substrate that is pulled through the interface. Patterned transfer occurs in a progressing linear front as the liquid layers recede. The process eliminates the dependency on gravity and sedimentation of prior methods, thereby extending the minimal chip size to the sub-100 micrometer scale. It provides a new route for the field of printable electronics to enable the integration of microscopic high performance inorganic semiconductors on foreign substrates with the freedom to choose target location, pitch, and integration density. As an example we demonstrate a fault-tolerant segmented flexible monocrystalline silicon solar cell, reducing the amount of Si that is used when compared to conventional rigid cells. PMID:20080682

  11. Femtosecond laser direct writing of monocrystalline hexagonal silver prisms

    SciTech Connect

    Vora, Kevin; Kang, SeungYeon; Moebius, Michael; Mazur, Eric

    2014-10-06

    Bottom-up growth methods and top-down patterning techniques are both used to fabricate metal nanostructures, each with a distinct advantage: One creates crystalline structures and the other offers precise positioning. Here, we present a technique that localizes the growth of metal crystals to the focal volume of a laser beam, combining advantages from both approaches. We report the fabrication of silver nanoprisms—hexagonal nanoscale silver crystals—through irradiation with focused femtosecond laser pulses. The growth of these nanoprisms is due to a nonlinear optical interaction between femtosecond laser pulses and a polyvinylpyrrolidone film doped with silver nitrate. The hexagonal nanoprisms have bases hundreds of nanometers in size and the crystal growth occurs over exposure times of less than 1 ms (8 orders of magnitude faster than traditional chemical techniques). Electron backscatter diffraction analysis shows that the hexagonal nanoprisms are monocrystalline. The fabrication method combines advantages from both wet chemistry and femtosecond laser direct-writing to grow silver crystals in targeted locations. The results presented in this letter offer an approach to directly positioning and growing silver crystals on a substrate, which can be used for plasmonic devices.

  12. Femtosecond laser direct writing of monocrystalline hexagonal silver prisms

    NASA Astrophysics Data System (ADS)

    Vora, Kevin; Kang, SeungYeon; Moebius, Michael; Mazur, Eric

    2014-10-01

    Bottom-up growth methods and top-down patterning techniques are both used to fabricate metal nanostructures, each with a distinct advantage: One creates crystalline structures and the other offers precise positioning. Here, we present a technique that localizes the growth of metal crystals to the focal volume of a laser beam, combining advantages from both approaches. We report the fabrication of silver nanoprisms—hexagonal nanoscale silver crystals—through irradiation with focused femtosecond laser pulses. The growth of these nanoprisms is due to a nonlinear optical interaction between femtosecond laser pulses and a polyvinylpyrrolidone film doped with silver nitrate. The hexagonal nanoprisms have bases hundreds of nanometers in size and the crystal growth occurs over exposure times of less than 1 ms (8 orders of magnitude faster than traditional chemical techniques). Electron backscatter diffraction analysis shows that the hexagonal nanoprisms are monocrystalline. The fabrication method combines advantages from both wet chemistry and femtosecond laser direct-writing to grow silver crystals in targeted locations. The results presented in this letter offer an approach to directly positioning and growing silver crystals on a substrate, which can be used for plasmonic devices.

  13. Whole-Genome Thermodynamic Analysis Reduces siRNA Off-Target Effects

    PubMed Central

    Chen, Xi; Liu, Peng; Chou, Hui-Hsien

    2013-01-01

    Small interfering RNAs (siRNAs) are important tools for knocking down targeted genes, and have been widely applied to biological and biomedical research. To design siRNAs, two important aspects must be considered: the potency in knocking down target genes and the off-target effect on any nontarget genes. Although many studies have produced useful tools to design potent siRNAs, off-target prevention has mostly been delegated to sequence-level alignment tools such as BLAST. We hypothesize that whole-genome thermodynamic analysis can identify potential off-targets with higher precision and help us avoid siRNAs that may have strong off-target effects. To validate this hypothesis, two siRNA sets were designed to target three human genes IDH1, ITPR2 and TRIM28. They were selected from the output of two popular siRNA design tools, siDirect and siDesign. Both siRNA design tools have incorporated sequence-level screening to avoid off-targets, thus their output is believed to be optimal. However, one of the sets we tested has off-target genes predicted by Picky, a whole-genome thermodynamic analysis tool. Picky can identify off-target genes that may hybridize to a siRNA within a user-specified melting temperature range. Our experiments validated that some off-target genes predicted by Picky can indeed be inhibited by siRNAs. Similar experiments were performed using commercially available siRNAs and a few off-target genes were also found to be inhibited as predicted by Picky. In summary, we demonstrate that whole-genome thermodynamic analysis can identify off-target genes that are missed in sequence-level screening. Because Picky prediction is deterministic according to thermodynamics, if a siRNA candidate has no Picky predicted off-targets, it is unlikely to cause off-target effects. Therefore, we recommend including Picky as an additional screening step in siRNA design. PMID:23484018

  14. Notch1 targeting siRNA delivery nanoparticles for rheumatoid arthritis therapy.

    PubMed

    Kim, Min Ju; Park, Jong-Sung; Lee, So Jin; Jang, Jiyeon; Park, Jin Su; Back, Seung Hyun; Bahn, Gahee; Park, Jae Hyung; Kang, Young Mo; Kim, Sun Hwa; Kwon, Ick Chan; Jo, Dong-Gyu; Kim, Kwangmeyung

    2015-10-28

    Notch pathway plays a pivotal role in synoviocytes involved in progression of rheumatoid arthritis (RA). Herein, we designed the Notch1 targeting siRNA delivery nanoparticles (siRNA-NPs) in order to confirm the anti-inflammatory effect in collagen-induced arthritis (CIA) model. The siRNA-NPs were successfully produced by encapsulating polymerized siRNA (poly-siRNA) into thiolated glycol chitosan (tGC) nanoparticles in aqueous condition. The in vitro Notch1 inhibition of siRNA-NPs in murine macrophage cell (RAW 264.7) was confirmed using confocal microscopy and real time PCR. Fluorescently labeled siRNA-NPs were successfully transfected in RAW 264.7 and modulated the expression of Notch1 in mRNA level. For in vivo study, siRNA-NPs exhibited the higher targeting efficiency in the arthritic joins of CIA mice, confirmed by the near-infrared fluorescence (NIRF) imaging. Furthermore, inhibition of Notch1 with siRNA-NPs resulted in retarded progression of inflammation, bone erosion, and cartilage damage in CIA mice. Novel Notch1 targeting siRNA delivery system of siRNA-NPs showed effective RA treatment by suppressing Notch1 signaling pathway without undesirable severe toxicity. Thus, Notch1 inhibiting siRNA-NPs demonstrated the great potential in RA therapeutics that was hard to be achieved using conventional drugs. Copyright © 2015 Elsevier B.V. All rights reserved.

  15. Cancer-targeting siRNA delivery from porous silicon nanoparticles.

    PubMed

    Wan, Yuan; Apostolou, Sinoula; Dronov, Roman; Kuss, Bryone; Voelcker, Nicolas H

    2014-10-01

    Porous silicon nanoparticles (pSiNPs) with tunable pore size are biocompatible and biodegradable, suggesting that they are suitable biomaterials as vehicles for drug delivery. Loading of small interfering RNA (siRNA) into the pores of pSiNPs can protect siRNA from degradation as well as improve the cellular uptake. We aimed to deliver MRP1 siRNA loaded into pSiNPs to glioblastoma cells, and to demonstrate downregulation of MRP1 at the mRNA and protein levels. 50-220 nm pSiNPs with an average pore size of 26 nm were prepared, followed by electrostatic adsorption of siRNA into pores. Oligonucleotide loading and release profiles were investigated; MRP1 mRNA and protein expression, cell viability and cell apoptosis were studied. Approximately 7.7 µg of siRNA was loaded per mg of pSiNPs. Cells readily took up nanoparticles after 30 min incubation. siRNA-loaded pSiNPs were able to effectively downregulate target mRNA (~40%) and protein expression (31%), and induced cell apoptosis and necrosis (33%). siRNA loaded pSiNPs downregulated mRNA and protein expression and induced cell death. This novel siRNA delivery system may pave the way towards developing more effective tumor therapies.

  16. Combined-environment influence on microcrack evolution in mono-crystalline silicon

    NASA Astrophysics Data System (ADS)

    Huang, W.-J.; Fortuno, Z. D.; Li, M.; Liu, J.; Liao, H.; Simmons-Potter, K.; Potter, B. G.

    2014-10-01

    The impact of combined environment conditions (mechanical state, temperature, and relative humidity) on microcrack propagation characteristics in p-type monocrystalline, photovoltaic-grade Si wafers was examined. A four-point bend apparatus was used to impose static strain conditions in 280 micron thick monocrystalline Si wafers containing microindentation-initiated crack centers. The specimen under test was simultaneously subjected to varied temperature and relative humidity conditions within a controlled environment chamber. Microcrack length was monitored after exposure to two sets of temperature and relative humidity conditions (i.e. 20° and 33%, 40° and 60% respectively) using scanning electron microscopy. Two primary stages of crack elongation behavior were observed under both of the combined environment conditions. Specifically, an early-time, more rapid growth period occurred, followed by more limited crack growth at later times. The deceleration of crack propagation is consistent with stress relaxation accompanying crack elongation under the constant strain conditions imposed. In general, an increase in the average microcrack propagation rate within both growth rate ranges and in the final overall change in average crack length was observed under elevated temperature and humidity conditions. These findings support the probable role of local crack-tip environment on microcrack evolution.

  17. Reducible hyaluronic acid-siRNA conjugate for target specific gene silencing.

    PubMed

    Park, Kitae; Yang, Jeong-A; Lee, Min-Young; Lee, Hwiwon; Hahn, Sei Kwang

    2013-07-17

    Despite wide applications of polymer-drug conjugates, there are only a few polymer-siRNA conjugates like poly(ethylene glycol) conjugated siRNA. In this work, reducible hyaluronic acid (HA)-siRNA conjugate was successfully developed for target specific systemic delivery of siRNA to the liver. The conjugation of siRNA to HA made it possible to form a compact nanocomplex of siRNA with relatively nontoxic linear polyethyleneimine (LPEI). After characterization of HA-siRNA conjugate by size exclusion chromatography (SEC) and gel electrophoresis, its complex formation with LPEI was investigated with a particle analyzer. The HA-siRNA/LPEI complex had a mean particle size of ca. 250 nm and a negative or neutral surface charge at physiological condition. The reducible HA-siRNA/LPEI complex showed a higher in vitro gene silencing efficiency than noncleavable HA-siRNA/LPEI complex. Furthermore, after systemic delivery, apolipoprotein B (ApoB) specific HA-siApoB/LPEI complex was target specifically delivered to the liver, which resulted in statistically significant reduction of ApoB mRNA expression in a dose dependent manner. The HA-siRNA conjugate can be effectively applied as a model system to the treatment of liver diseases using various siRNAs and relatively nontoxic polycations.

  18. Delivery of siRNA into breast cancer cells via phage fusion protein-targeted liposomes.

    PubMed

    Bedi, Deepa; Musacchio, Tiziana; Fagbohun, Olusegun A; Gillespie, James W; Deinnocentes, Patricia; Bird, R Curtis; Bookbinder, Lonnie; Torchilin, Vladimir P; Petrenko, Valery A

    2011-06-01

    Efficacy of siRNAs as potential anticancer therapeutics can be increased by their targeted delivery into cancer cells via tumor-specific ligands. Phage display offers a unique approach to identify highly specific and selective ligands that can deliver nanocarriers to the site of disease. In this study, we proved a novel approach for intracellular delivery of siRNAs into breast cancer cells through their encapsulation into liposomes targeted to the tumor cells with preselected intact phage proteins. The targeted siRNA liposomes were obtained by a fusion of two parental liposomes containing spontaneously inserted siRNA and fusion phage proteins. The presence of pVIII coat protein fused to a MCF-7 cell-targeting peptide DMPGTVLP in the liposomes was confirmed by Western blotting. The novel phage-targeted siRNA-nanopharmaceuticals demonstrate significant down-regulation of PRDM14 gene expression and PRDM14 protein synthesis in the target MCF-7 cells. This approach offers the potential for development of new anticancer siRNA-based targeted nanomedicines. In this study, the authors report a novel approach for targeted intracellular delivery of siRNAs into breast cancer cells through encapsulation into liposomes targeted to the tumor cells with preselected intact phage proteins. Copyright © 2011 Elsevier Inc. All rights reserved.

  19. A Nonpolycationic Fully Proteinaceous Multiagent System for Potent Targeted Delivery of siRNA

    PubMed Central

    Liu, David V; Yang, Nicole J; Wittrup, K Dane

    2014-01-01

    Protein-based methods of targeted short-interfering RNA (siRNA) delivery have the potential to solve some of the problems faced by nanoparticle-based methods, such as poor pharmacokinetics and biodistribution, low tumor penetration, and polydispersity. However, protein-based targeted delivery has been limited to fusion proteins with polycationic peptides as siRNA carriers, whose high charge density in some cases results in undesirable biophysical and in vivo properties. Here, we present a fully proteinaceous, multiagent approach for targeted siRNA delivery to epidermal growth factor receptor (EGFR), using a nonpolycationic carrier for siRNA. Each agent contributes a fundamentally different mechanism of action that work together for potent targeted RNA interference. The first agent is an EGFR-targeted fusion protein that uses a double-stranded RNA-binding domain as a nonpolycationic siRNA carrier. This double-stranded RNA-binding domain fusion protein can deliver siRNA to the endosomes of an EGFR-expressing cell line. A second agent delivers the cholesterol-dependent cytolysin, perfringolysin O, in a targeted manner, which enhances the endosomal escape of siRNA and induces gene silencing. A third agent that clusters EGFR increases gene-silencing potency and decreases cytolysin toxicity. Altogether, this system is potent, with only 16 nmol/l siRNA required for gene silencing and a therapeutic window that spans two orders of magnitude of targeted cytolysin concentrations. PMID:24825362

  20. Kinetic analysis of the effects of target structure on siRNA efficiency

    NASA Astrophysics Data System (ADS)

    Chen, Jiawen; Zhang, Wenbing

    2012-12-01

    RNAi efficiency for target cleavage and protein expression is related to the target structure. Considering the RNA-induced silencing complex (RISC) as a multiple turnover enzyme, we investigated the effect of target mRNA structure on siRNA efficiency with kinetic analysis. The 4-step model was used to study the target cleavage kinetic process: hybridization nucleation at an accessible target site, RISC-mRNA hybrid elongation along with mRNA target structure melting, target cleavage, and enzyme reactivation. At this model, the terms accounting for the target accessibility, stability, and the seed and the nucleation site effects are all included. The results are in good agreement with that of experiments which show different arguments about the structure effects on siRNA efficiency. It shows that the siRNA efficiency is influenced by the integrated factors of target's accessibility, stability, and the seed effects. To study the off-target effects, a simple model of one siRNA binding to two mRNA targets was designed. By using this model, the possibility for diminishing the off-target effects by the concentration of siRNA was discussed.

  1. Picosecond optical vortex pulse illumination forms a monocrystalline silicon needle.

    PubMed

    Takahashi, Fuyuto; Miyamoto, Katsuhiko; Hidai, Hirofumi; Yamane, Keisaku; Morita, Ryuji; Omatsu, Takashige

    2016-02-24

    The formation of a monocrystalline silicon needle by picosecond optical vortex pulse illumination was demonstrated for the first time in this study. The dynamics of this silicon needle formation was further revealed by employing an ultrahigh-speed camera. The melted silicon was collected through picosecond pulse deposition to the dark core of the optical vortex, forming the silicon needle on a submicrosecond time scale. The needle was composed of monocrystalline silicon with the same lattice index (100) as that of the silicon substrate, and had a height of approximately 14 μm and a thickness of approximately 3 μm. Overlaid vortex pulses allowed the needle to be shaped with a height of approximately 40 μm without any changes to the crystalline properties. Such a monocrystalline silicon needle can be applied to devices in many fields, such as core-shell structures for silicon photonics and photovoltaic devices as well as nano- or microelectromechanical systems.

  2. Monocrystalline test structures, and use for calibrating instruments

    DOEpatents

    Cresswell, Michael W.; Ghoshtagore, R. N.; Linholm, Loren W.; Allen, Richard A.; Sniegowski, Jeffry J.

    1997-01-01

    An improved test structure for measurement of width of conductive lines formed on substrates as performed in semiconductor fabrication, and for calibrating instruments for such measurements, is formed from a monocrystalline starting material, having an insulative layer formed beneath its surface by ion implantation or the equivalent, leaving a monocrystalline layer on the surface. The monocrystalline surface layer is then processed by preferential etching to accurately define components of the test structure. The substrate can be removed from the rear side of the insulative layer to form a transparent window, such that the test structure can be inspected by transmissive-optical techniques. Measurements made using electrical and optical techniques can be correlated with other measurements, including measurements made using scanning probe microscopy.

  3. Structure-Guided Control of siRNA Off-Target Effects.

    PubMed

    Suter, Scott R; Sheu-Gruttadauria, Jessica; Schirle, Nicole T; Valenzuela, Rachel; Ball-Jones, Alexi A; Onizuka, Kazumitsu; MacRae, Ian J; Beal, Peter A

    2016-07-20

    Short interfering RNAs (siRNAs) are promising therapeutics that make use of the RNA interference (RNAi) pathway, but liabilities arising from the native RNA structure necessitate chemical modification for drug development. Advances in the structural characterization of components of the human RNAi pathway have enabled structure-guided optimization of siRNA properties. Here we report the 2.3 Å resolution crystal structure of human Argonaute 2 (hAgo2), a key nuclease in the RNAi pathway, bound to an siRNA guide strand bearing an unnatural triazolyl nucleotide at position 1 (g1). Unlike natural nucleotides, this analogue inserts deeply into hAgo2's central RNA binding cleft and thus is able to modulate pairing between guide and target RNAs. The affinity of the hAgo2-siRNA complex for a seed-only matched target was significantly reduced by the triazolyl modification, while the affinity for a fully matched target was unchanged. In addition, siRNA potency for off-target repression was reduced (4-fold increase in IC50) by the modification, while on-target knockdown was improved (2-fold reduction in IC50). Controlling siRNA on-target versus microRNA (miRNA)-like off-target potency by projection of substituent groups into the hAgo2 central cleft from g1 is a new approach to enhance siRNA selectivity with a strong structural rationale.

  4. Topics in LIFE Target Survival: 11-SI-004 Final Report

    SciTech Connect

    Miles, Robin; Benett, Bill; Bond, Tiziana; Chang, Allan; Dawson, David; Fornasiero, Francesco; Hamilton, Julie; Havstad, Mark; Kucheyev, Sergei; LeBlanc, Mary; Rosso, Paul; Schebler, Greg; Van Cleve, Eli; Worsley, Marcus

    2014-10-20

    The LIFE target design incorporates many considerations to generate the desired fusion gain including the physics design, the cost of manufacturing of the target, the injectability of the target, the aerodynamic flight characteristics of the target, the ability to track and engage the target and to maintain the structural and thermal integrity of the target. This document describes the effort that was made in support of issues of survivability of the target during injection which included issues massmanufactural materials and processes which could be used in the target.

  5. RGD-based active targeting of novel polycation liposomes bearing siRNA for cancer treatment.

    PubMed

    Yonenaga, Norihito; Kenjo, Eriya; Asai, Tomohiro; Tsuruta, Atsushi; Shimizu, Kosuke; Dewa, Takehisa; Nango, Mamoru; Oku, Naoto

    2012-06-10

    For the purpose of systemic delivery of siRNA, we previously developed polycation liposomes (PCLs) containing dicetylphosphate-tetraethylenepentamine (DCP-TEPA) as an effective siRNA carrier. In the present study, to endow these PCLs (TEPA-PCL) actively target cancer cells and angiogenic vessels, we decorated the PCLs with cyclic RGD, by using cyclic RGD-grafted distearoylphosphatidylethanolamine-polyethylene glycol (DSPE-PEG), and investigated the usefulness of this type of carrier (RGD-PEG-PCL) for active targeting. Firstly, the gene-silencing efficacy of siRNA for luciferase (siLuc2) formulated in RGD-PEG-PCL (RGD-PEG-PCL/siLuc2) was examined in vitro by using B16F10-luc2 murine melanoma cells stably expressing the luciferase 2 gene, where the siRNA was grafted with cholesterol at the 3'-end of the sense strand (siRNA-C) for the stable association of the siRNA with the PCL. RGD-PEG-PCL/siLuc2 showed high knockdown efficiency compared with siLuc2 formulated in PEGylated TEPA-PCL without cyclic RGD (PEG-PCL). Next, the gene-silencing efficacy of RGD-PEG-PCL/siLuc2 was examined in vivo by use of B16F10-luc2 lung metastatic model mice. The intravenous injection of RGD-PEG-PCL/siLuc2 showed high knockdown efficiency against metastatic B16F10-luc2 tumors in the lungs of the mice, as assessed with an in vivo imaging system. These data strongly suggest that systemic and active targeting siRNA delivery using RGD-PEG-PCL is useful for cancer RNAi therapy.

  6. Targeted Delivery of Anti-coxsackievirus siRNAs Using Ligand-conjugated Packaging RNAs

    PubMed Central

    Zhang, Huifang M.; Su, Yue; Guo, Songchuan; Yuan, Ji; Lim, Travis; Liu, Jing; Guo, Peixuan; Yang, Decheng

    2013-01-01

    Coxsackievirus B3 (CVB3) is a common pathogen of myocarditis. We previously synthesized a siRNA targeting the CVB3 protease 2A (siRNA/2A) gene and achieved reduction of CVB3 replication by 92% in vitro. However, like other drugs under development, CVB3 siRNA faces a major challenge of targeted delivery. In this study, we investigated a novel approach to deliver CVB3 siRNAs to a specific cell population (e.g. HeLa cells containing folate receptor) using receptor ligand (folate)-linked packaging RNA (pRNA) from bacterial phage phi29. pRNA monomers can spontaneously form dimers and multimers under optimal conditions by base-pairing between their stem loops. By covalently linking a fluorescence-tag to folate, we delivered the conjugate specifically to HeLa cells without the need of transfection. We further demonstrated that pRNA covalently conjugated to siRNA/2A achieved an equivalent antiviral effect to that of the siRNA/2A alone. Finally, the drug targeted delivery was further evaluated by using pRNA monomers or dimers, which carried both the siRNA/2A and folate ligand and demonstrated that both of them strongly inhibited CVB3 replication. These data indicate that pRNA as a siRNA carrier can specifically deliver the drug to target cells via its ligand and specific receptor interaction and inhibit virus replication effectively. PMID:19616030

  7. DELIVERY OF siRNA INTO BREAST CANCER CELLS VIA PHAGE FUSION PROTEIN-TARGETED LIPOSOMES

    PubMed Central

    Bedi, Deepa; Musacchio, Tiziana; Fagbohun, Olusegun A.; Gillespie, James W.; Deinnocentes, Patricia; Bird, R. Curtis; Bookbinder, Lonnie; Torchilin, Vladimir P.; Petrenko, Valery A.

    2011-01-01

    Efficacy of siRNAs as potential anticancer therapeutics can be increased by their targeted delivery into cancer cells via tumor-specific ligands. Phage display offers an unique approach to identify highly specific and selective ligands that can deliver nanocarriers to the site of disease. In this study, we proved a novel approach for intracellular delivery of siRNAs into breast cancer cells through their encapsulation into liposomes targeted to the tumor cells with preselected intact phage proteins. The targeted siRNA liposomes were obtained by a fusion of two parental liposomes containing spontaneously inserted siRNA and fusion phage proteins. The presence of pVIII coat protein fused to a MCF-7 cell-targeting peptide DMPGTVLP in the liposomes was confirmed by Western blotting. The novel phage-targeted siRNA-nanopharmaceuticals demonstrate significant down-regulation of PRDM14 gene expression and PRDM14 protein synthesis in the target MCF- 7 cells. This approach offers the potential for development of new anticancer siRNA-based targeted nanomedicines. PMID:21050894

  8. Effect of crystal plane orientation on tribochemical removal of monocrystalline silicon

    NASA Astrophysics Data System (ADS)

    Xiao, Chen; Guo, Jian; Zhang, Peng; Chen, Cheng; Chen, Lei; Qian, Linmao

    2017-01-01

    The effect of crystal plane orientation on tribochemical removal of monocrystalline silicon was investigated using an atomic force microscope. Experimental results indicated that the tribochemical removal of silicon by SiO2 microsphere presented strong crystallography-induced anisotropy. Further analysis suggested that such anisotropic tribochemical removal of silicon was not dependent on the crystallography-dependent surface mechanical properties (i.e., hardness and elastic modulus), but was mainly attributed to various atomic planar density and interplanar spacing in different crystal planes. Phenomenological results speculated that higher density of silicon atom could promote the formation of Si-O-Si bonds between the SiO2 microsphere and silicon substrate, resulting in more severe tribochemical material removal. Larger interplanar spacing with smaller energy barrier facilitated the rupture of the Si-Si network with the help of mechanical shearing stress, which caused more serious wear of the silicon surface. The results may help understand the material removal mechanism of silicon and provide useful knowledge for chemical mechanical polishing.

  9. Effect of crystal plane orientation on tribochemical removal of monocrystalline silicon

    PubMed Central

    Xiao, Chen; Guo, Jian; Zhang, Peng; Chen, Cheng; Chen, Lei; Qian, Linmao

    2017-01-01

    The effect of crystal plane orientation on tribochemical removal of monocrystalline silicon was investigated using an atomic force microscope. Experimental results indicated that the tribochemical removal of silicon by SiO2 microsphere presented strong crystallography-induced anisotropy. Further analysis suggested that such anisotropic tribochemical removal of silicon was not dependent on the crystallography-dependent surface mechanical properties (i.e., hardness and elastic modulus), but was mainly attributed to various atomic planar density and interplanar spacing in different crystal planes. Phenomenological results speculated that higher density of silicon atom could promote the formation of Si-O-Si bonds between the SiO2 microsphere and silicon substrate, resulting in more severe tribochemical material removal. Larger interplanar spacing with smaller energy barrier facilitated the rupture of the Si-Si network with the help of mechanical shearing stress, which caused more serious wear of the silicon surface. The results may help understand the material removal mechanism of silicon and provide useful knowledge for chemical mechanical polishing. PMID:28084433

  10. Thick target yields of proton induced gamma-ray emission from Al, Si and P

    NASA Astrophysics Data System (ADS)

    Jokar, A.; Kakuee, O.; Lamehi-Rachti, M.; Fathollahi, V.

    2017-03-01

    Thick target excitation yield curves of gamma-rays from the reactions 27Al(p,p‧γ)27Al (Eγ = 844 and 1014 keV), 27Al(p,αγ)27Al (Eγ = 1369 keV), 28Si(p,p‧γ)28Si (Eγ = 1779 keV), 29Si(p,p‧γ)29Si (Eγ = 1273 keV) and 31P(p,p‧γ)31P (Eγ = 1266 keV) were measured by bombarding pure-element targets with protons at energies below 3 MeV. Gamma-rays were detected with a High Purity Ge detector placed at an angle of 90° with respect to the beam direction. The obtained thick target gamma-ray yields were compared with the previously published data. The overall systematic uncertainty of the thick target yield values was estimated to be better than ±9%.

  11. Polarization effects in femtosecond laser induced amorphization of monocrystalline silicon

    NASA Astrophysics Data System (ADS)

    Bai, Feng; Li, Hong-Jin; Huang, Yuan-Yuan; Fan, Wen-Zhong; Pan, Huai-Hai; Wang, Zhuo; Wang, Cheng-Wei; Qian, Jing; Li, Yang-Bo; Zhao, Quan-Zhong

    2016-10-01

    We have used femtosecond laser pulses to ablate monocrystalline silicon wafer. Raman spectroscopy and X-ray diffraction analysis of ablation surface indicates horizontally polarized laser beam shows an enhancement in amorphization efficiency by a factor of 1.6-1.7 over the circularly polarized laser ablation. This demonstrates that one can tune the amorphization efficiency through the polarization of irradiation laser.

  12. Ultrasound-Targeted Microbubble Destruction to Deliver siRNA Cancer Therapy

    PubMed Central

    Carson, Andrew R; McTiernan, Charles F; Lavery, Linda; Grata, Michelle; Leng, Xiaoping; Wang, Jianjun; Chen, Xucai; Villanueva, Flordeliza S

    2012-01-01

    Microbubble contrast agents can specifically deliver nucleic acids to target tissues when exposed to ultrasound treatment parameters that mediate microbubble destruction. In this study, we evaluated whether microbubbles and ultrasound targeted microbubble destruction (UTMD) could be used to enhance delivery of EGFR-directed small inhibitory RNA (siRNA) to murine squamous cell carcinomas. Custom designed microbubbles efficiently bound siRNA and mediated RNAse protection. UTMD-mediated delivery of microbubbles loaded with EGFR-directed siRNA to murine squamous carcinoma cells in vitro reduced EGFR expression and EGF-dependent growth, relative to delivery of control siRNA. Similarly, serial UTMD-mediated delivery of EGFR siRNA to squamous cell carcinoma in vivo decreased EGFR expression and increased tumor doubling times, relative to controls receiving EGFR siRNA loaded microbubbles but not ultrasound or control siRNA loaded microbubbles and UTMD. Taken together, our results offer a preclinical proof of concept for customized microbubbles and UTMD to deliver gene-targeted siRNA for cancer therapy. PMID:23010078

  13. Exosomes for targeted siRNA delivery across biological barriers.

    PubMed

    El Andaloussi, Samir; Lakhal, Samira; Mäger, Imre; Wood, Matthew J A

    2013-03-01

    Using oligonucleotide-based drugs to modulate gene expression has opened a new avenue for drug discovery. In particular small interfering RNAs (siRNAs) are being rapidly recognized as promising therapeutic tools, but their poor bioavailability limits the full realization of their clinical potential. In recent years, cumulating evidence has emerged for the role of membrane vesicles, secreted by most cells and found in all body fluids, as key mediators of information transmission between cells. Importantly, a sub-group of these termed exosomes, have recently been shown to contain various RNA species and to mediate their horizontal transfer to neighbouring- or distant recipient cells. Here, we provide a brief overview on membrane vesicles and their role in exchange of genetic information. We also describe how these natural carriers of genetic material can be harnessed to overcome the obstacle of poor delivery and allow efficient systemic delivery of exogenous siRNA across biological barriers such as the blood-brain barrier.

  14. ICS-283: a system for targeted intravenous delivery of siRNA.

    PubMed

    Schiffelers, Raymond M; Storm, Gert

    2006-05-01

    ICS-283 was developed within Intradigm Corporation as a system that is designed for the systemic delivery of therapeutic small interfering (siRNA) to sites of pathological angiogenesis. The non-viral siRNA delivery system is based on synthetic nanoparticles, known as Targe (Intradigm Corporation), which functions as a broad-platform technology to deliver siRNA to specific target cells in diseased tissues. The system is constructed to incorporate different functionalities that address critical needs for successful nucleic acid delivery. The TargeTran synthetic vector is a self-assembling, layered nanoparticle that protects and targets siRNA to specific cell types in pathological tissues. At present, ICS-283 is the only antiangiogenic siRNA delivery system that is designed for intravenous administration to treat angiogenesis-driven diseases.

  15. Targeted Sterically Stabilized Phospholipid siRNA Nanomedicine for Hepatic and Renal Fibrosis

    PubMed Central

    Khaja, Fatima; Jayawardena, Dulari; Kuzmis, Antonina; Önyüksel, Hayat

    2016-01-01

    Since its discovery, small interfering RNA (siRNA) has been considered a potent tool for modulating gene expression. It has the ability to specifically target proteins via selective degradation of messenger RNA (mRNA) not easily accessed by conventional drugs. Hence, RNA interference (RNAi) therapeutics have great potential in the treatment of many diseases caused by faulty protein expression such as fibrosis and cancer. However, for clinical application siRNA faces a number of obstacles, such as poor in vivo stability, and off-target effects. Here we developed a unique targeted nanomedicine to tackle current siRNA delivery issues by formulating a biocompatible, biodegradable and relatively inexpensive nanocarrier of sterically stabilized phospholipid nanoparticles (SSLNPs). This nanocarrier is capable of incorporating siRNA in its core through self-association with a novel cationic lipid composed of naturally occuring phospholipids and amino acids. This overall assembly protects and delivers sufficient amounts of siRNA to knockdown over-expressed protein in target cells. The siRNA used in this study, targets connective tissue growth factor (CTGF), an important regulator of fibrosis in both hepatic and renal cells. Furthermore, asialoglycoprotein receptors are targeted by attaching the galactosamine ligand to the nanocarries which enhances the uptake of nanoparticles by hepatocytes and renal tubular epithelial cells, the major producers of CTGF in fibrosis. On animals this innovative nanoconstruct, small interfering RNA in sterically stabilized phospholipid nanoparticles (siRNA-SSLNP), showed favorable pharmacokinetic properties and accumulated mostly in hepatic and renal tissues making siRNA-SSLNP a suitable system for targeting liver and kidney fibrotic diseases.

  16. Construction of a PLGA based, targeted siRNA delivery system for treatment of osteoporosis.

    PubMed

    Sezlev Bilecen, Deniz; Rodriguez-Cabello, Jose Carlos; Uludag, Hasan; Hasirci, Vasif

    2017-11-01

    Osteoporosis, a systemic skeletal disorder, occurs when bone turnover balance is disrupted. With the identification of the genes involved in the pathogenesis of the disease, studies on development of new treatments has intensified. Short interfering RNA (siRNA) is used to knockdown disease related gene expressions. Targeting siRNA in vivo is challenging. The maintenance of therapeutic plasma level is hampered by clearance of siRNA from the body. Targeted systems are useful in increasing the drug concentration at the target site and decreasing side effects. Aim of the present study was to develop an injectable siRNA delivery system to protect siRNA during systemic distribution and target the siRNA to bone tissue using a thermoresponsive, genetically engineered, elastin-like recombinamer (ELR), designed to interact with the mineral component of bone. The delivery system consisted of DNAoligo as a siRNA substitute complexed with the cationic polymer, polyethyleneimine (PEI), at N/P ratio of 20. The complex was encapsulated in poly(lactic acid-co-glycolic acid) (PLGA) nanocapsules. PLGA capsules were characterized by SEM, TEM and XPS. FTIR was used to show the preferential attachment of ELR to HAp. Encapsulation efficiency of the complex in PLGA nanocapsules was 48%. The release kinetics of the complex fits the Higuchi release kinetics.

  17. [siRNAs with high specificity to the target: a systematic design by CRM algorithm].

    PubMed

    Alsheddi, T; Vasin, L; Meduri, R; Randhawa, M; Glazko, G; Baranova, A

    2008-01-01

    'Off-target' silencing effect hinders the development of siRNA-based therapeutic and research applications. Common solution to this problem is an employment of the BLAST that may miss significant alignments or an exhaustive Smith-Waterman algorithm that is very time-consuming. We have developed a Comprehensive Redundancy Minimizer (CRM) approach for mapping all unique sequences ("targets") 9-to-15 nt in size within large sets of sequences (e.g. transcriptomes). CRM outputs a list of potential siRNA candidates for every transcript of the particular species. These candidates could be further analyzed by traditional "set-of-rules" types of siRNA designing tools. For human, 91% of transcripts are covered by candidate siRNAs with kernel targets of N = 15. We tested our approach on the collection of previously described experimentally assessed siRNAs and found that the correlation between efficacy and presence in CRM-approved set is significant (r = 0.215, p-value = 0.0001). An interactive database that contains a precompiled set of all human siRNA candidates with minimized redundancy is available at http://129.174.194.243. Application of the CRM-based filtering minimizes potential "off-target" silencing effects and could improve routine siRNA applications.

  18. Ocular neuroprotection by siRNA targeting caspase-2

    PubMed Central

    Ahmed, Z; Kalinski, H; Berry, M; Almasieh, M; Ashush, H; Slager, N; Brafman, A; Spivak, I; Prasad, N; Mett, I; Shalom, E; Alpert, E; Di Polo, A; Feinstein, E; Logan, A

    2011-01-01

    Retinal ganglion cell (RGC) loss after optic nerve damage is a hallmark of certain human ophthalmic diseases including ischemic optic neuropathy (ION) and glaucoma. In a rat model of optic nerve transection, in which 80% of RGCs are eliminated within 14 days, caspase-2 was found to be expressed and cleaved (activated) predominantly in RGC. Inhibition of caspase-2 expression by a chemically modified synthetic short interfering ribonucleic acid (siRNA) delivered by intravitreal administration significantly enhanced RGC survival over a period of at least 30 days. This exogenously delivered siRNA could be found in RGC and other types of retinal cells, persisted inside the retina for at least 1 month and mediated sequence-specific RNA interference without inducing an interferon response. Our results indicate that RGC apoptosis induced by optic nerve injury involves activation of caspase-2, and that synthetic siRNAs designed to inhibit expression of caspase-2 represent potential neuroprotective agents for intervention in human diseases involving RGC loss. PMID:21677688

  19. CXCR4-targeted modular peptide carriers for efficient anti-VEGF siRNA delivery.

    PubMed

    Egorova, Anna; Shubina, Anastasia; Sokolov, Dmitriy; Selkov, Sergey; Baranov, Vladislav; Kiselev, Anton

    2016-12-30

    The application of small interfering RNA (siRNA) for specific gene inhibition is a promising strategy in gene therapy treatments. The efficient cellular delivery of therapeutic siRNA is a critical step in RNA interference (RNAi) application. Highly efficient siRNA carriers should be developed for specific cellular uptake, stable RNA-complexes formation and intracellular RNA release. To study these features, we evaluated modular peptide carriers bearing CXCR4 targeting ligand for their ability to condense siRNA, facilitate endosomal escape and VEGFA gene silencing in CXCR4-expressing endothelial and glioblastoma cells. Peptide carriers were shown to condense and protect siRNA from RNAse degradation. Various N/P ratios were used for physicochemical characterization to optimize siRNA/peptide complexes for in vitro studies. On average, cytotoxicity of siRNA-polyplexes depended on cell type and was not higher than that of PEI/siRNA complexes. VEGFA gene knockdown was significantly improved with CXCR4-targeted carriers in contrast to nontargeted peptides. siRNA delivery by means of ligandconjugated carriers resulted in 2.5-3-fold decrease of VEGF expression in glioblastoma cells and in 1.5-2-fold decrease of VEGF expression in endothelial cells. Delivery of siRNA/peptide complexes resulted in 2-6- fold decrease in VEGF protein yield and in significant inhibition of endothelial cells migration. The study shows that implication of peptide carriers modified with CXCR4 ligand is a promising approach to develop targeted siRNA delivery system into CXCR4-expressing cancer and endothelial cells. Copyright © 2016 Elsevier B.V. All rights reserved.

  20. Selecting effective siRNA target sequences by using Bayes' theorem.

    PubMed

    Takasaki, Shigeru

    2009-10-01

    Short interfering RNA (siRNA) has been widely used for studying gene functions in mammalian cells but varies markedly in its gene silencing efficacy. Although many design rules/guidelines for effective siRNAs based on various criteria have been reported recently, there are few consistencies among them. This makes it difficult to select effective siRNA sequences in mammalian genes. Another shortcoming of most previously reported methods is that they cannot estimate the probability that a candidate sequence will silence the target gene. The analytical prediction method proposed in the present study uses Bayes' theorem to select effective siRNA target sequences from many possible candidate sequences. It is quite different from the previous score-based siRNA design techniques and can predict the probability that a candidate siRNA sequence will be effective. The results of evaluating it by applying it to recently reported effective and ineffective siRNA sequences for various genes indicate that it would be useful for many other genes. It should therefore be useful for selecting siRNA sequences effective for mammalian genes.

  1. Targeted delivery of siRNA into breast cancer cells via phage fusion proteins.

    PubMed

    Bedi, Deepa; Gillespie, James W; Petrenko, Vasily A; Ebner, Andreas; Leitner, Michael; Hinterdorfer, Peter; Petrenko, Valery A

    2013-02-04

    Nucleic acids, including antisense oligonucleotides, small interfering RNA (siRNA), aptamers, and rybozymes, emerged as versatile therapeutics due to their ability to interfere in a well-planned manner with the flow of genetic information from DNA to protein. However, a systemic use of NAs is hindered by their instability in physiological liquids and inability of intracellular accumulation in the site of action. We first evaluated the potential of cancer specific phage fusion proteins as targeting ligands that provide encapsulation, protection, and navigation of siRNA to the target cell. The tumor-specific proteins were isolated from phages that were affinity selected from a landscape phage library against target breast cancer cells. It was found that fusion phage coat protein fpVIII displaying cancer-targeting peptides can effectively encapsulate siRNAs and deliver them into the cells leading to specific silencing of the model gene GAPDH. Complexes of siRNA and phage protein form nanoparticles (nanophages), which were characterized by atomic force microscopy and ELISA, and their stability was demonstrated by resistance of encapsulated siRNA to degradation by serum nucleases. The phage protein/siRNA complexes can make a new type of highly selective, stable, active, and physiologically acceptable cancer nanomedicine.

  2. Target-specific delivery of siRNA into hepatoma cells' cytoplasm by bifunctional carrier peptide.

    PubMed

    Liu, Xiaoxuan; Zhu, Lin; Ma, Jingjing; Qiao, Xinxiao; Zhu, Dunwan; Liu, Lanxia; Leng, Xigang

    2017-02-01

    RNA interference (RNAi) is among the most potential approach for the therapy of hepatocellular carcinoma and the major barrier hindering siRNA therapeutics is the low efficiency of delivery to the desired cells. The current study aimed at developing a novel peptide for more efficient hepatoma targeted siRNA delivery, by combining luteinizing hormone-releasing hormone with hepatoma targeting specificity and MPG(△NLS) with cytoplasm-delivery tendency. The developed bifunctional peptide LHRH-MPG(△NLS) and siRNA were mixed together and resulted in LHRH-MPG(△NLS)/siRNA polyplexes through self-assembly. The polyplexes were characterized by agarose gel retardation and dynamic light scatting analysis. Hepatoma targeting specificity was analyzed with the GE IN Cell Analyzer 2000 High-Content Cellular Analysis System after cell transfection, and the effect of RNA interference was detected by RT-PCR. The results demonstrated that LHRH-MPG(△NLS) was able to assemble with siRNA to form stable and nano-sized peptide/siRNA polyplexes, which could inhibit the expression of the target gene and was essentially non-cytotoxic, as compared with the commercial transfection reagent lipofectamine 2000.

  3. Targeted delivery of Dicer-substrate siRNAs using a dual targeting peptide decorated dendrimer delivery system.

    PubMed

    Liu, Xiaoxuan; Liu, Cheng; Chen, Chao; Bentobji, Mélanie; Cheillan, Francine Azario; Piana, Jeanne Thomassin; Qu, Fanqi; Rocchi, Palma; Peng, Ling

    2014-11-01

    Small interfering RNAs (siRNA) are emerging as novel therapeutic agents, providing competent delivery systems that are available. Dendrimers, a special family of synthetic macromolecules, represent an exciting delivery platform by virtue of their well-defined dendritic structure and unique multivalency and cooperativity confined within a nanoscale volume. Here, we report a Dicer-substrate siRNA (dsiRNA) which, when delivered using a structurally flexible triethanolamine-core poly(amidoamine) dendrimer of generation 5 as the nanocarrier, gives rise to a much greater RNAi response than that produced with conventional siRNA. Further decoration of the dsiRNA/dendrimer complexes with a dual targeting peptide simultaneously promoted cancer cell targeting through interacting with integrins and cell penetration via the interaction with neuropilin-1 receptors, which led to improved gene silencing and anticancer activity. Altogether, our results disclosed here open a new avenue for therapeutic implementation of RNAi using dendrimer nanovector based targeted delivery. This study demonstrates superior therapeutic properties of siRNA when combined with a dendrimer-based targeted nano-delivery system. Similar approaches may eventually gain clinical utility following additional studies determining safety and efficacy. Copyright © 2014 Elsevier Inc. All rights reserved.

  4. Tumor-targeted delivery of siRNA using fatty acyl-CGKRK peptide conjugates.

    PubMed

    Sharma, Meenakshi; El-Sayed, Naglaa Salem; Do, Hung; Parang, Keykavous; Tiwari, Rakesh Kumar; Aliabadi, Hamidreza Montazeri

    2017-07-21

    Tumor-targeted carriers provide efficient delivery of chemotherapeutic agents to tumor tissue. CGKRK is one of the well-known tumor targeting peptides with significant specificity for angiogenic blood vessels and tumor cells. Here, we designed fatty acyl conjugated CGKRK peptides, based on the hypothesis that hydrophobically-modified CGKRK peptide could enhance cellular permeation and delivery of siRNA targeted to tumor cells for effective silencing of selected proteins. We synthesized six fatty acyl-peptide conjugates, using a diverse chain of saturated and unsaturated fatty acids to study the efficiency of this approach. At peptide:siRNA weight/weight ratio of 10:1 (N/P ≈ 13.6), almost all the peptides showed complete binding with siRNA, and at a w/w ratio of 20:1 (N/P ≈ 27.3), complete protection of siRNA from early enzymatic degradation was observed. Conjugated peptides and peptide/siRNA complexes did not show significant cytotoxicity in selected cell lines. The oleic acid-conjugated peptide showed the highest efficiency in siRNA uptake and silencing of kinesin spindle protein at peptide:siRNA w/w ratio of 80:1 (N/P ≈ 109). The siRNA internalization into non-tumorigenic kidney cells was negligible with all fatty acyl-peptide conjugates. These results indicate that conjugation of fatty acids to CGKRK could create an efficient delivery system for siRNA silencing specifically in tumor cells.

  5. Mechanical instability of monocrystalline and polycrystalline methane hydrates

    PubMed Central

    Wu, Jianyang; Ning, Fulong; Trinh, Thuat T.; Kjelstrup, Signe; Vlugt, Thijs J. H.; He, Jianying; Skallerud, Bjørn H.; Zhang, Zhiliang

    2015-01-01

    Despite observations of massive methane release and geohazards associated with gas hydrate instability in nature, as well as ductile flow accompanying hydrate dissociation in artificial polycrystalline methane hydrates in the laboratory, the destabilising mechanisms of gas hydrates under deformation and their grain-boundary structures have not yet been elucidated at the molecular level. Here we report direct molecular dynamics simulations of the material instability of monocrystalline and polycrystalline methane hydrates under mechanical loading. The results show dislocation-free brittle failure in monocrystalline hydrates and an unexpected crossover from strengthening to weakening in polycrystals. Upon uniaxial depressurisation, strain-induced hydrate dissociation accompanied by grain-boundary decohesion and sliding destabilises the polycrystals. In contrast, upon compression, appreciable solid-state structural transformation dominates the response. These findings provide molecular insight not only into the metastable structures of grain boundaries, but also into unusual ductile flow with hydrate dissociation as observed during macroscopic compression experiments. PMID:26522051

  6. Hollow carbon nanobubbles: monocrystalline MOF nanobubbles and their pyrolysis.

    PubMed

    Zhang, Wei; Jiang, Xiangfen; Zhao, Yanyi; Carné-Sánchez, Arnau; Malgras, Victor; Kim, Jeonghun; Kim, Jung Ho; Wang, Shaobin; Liu, Jian; Jiang, Ji-Sen; Yamauchi, Yusuke; Hu, Ming

    2017-05-01

    While bulk-sized metal-organic frameworks (MOFs) face limits to their utilization in various research fields such as energy storage applications, nanoarchitectonics is believed to be a possible solution. It is highly challenging to realize MOF nanobubbles with monocrystalline frameworks. By a spatially controlled etching approach, here, we can achieve the synthesis of zeolitic imidazolate framework (ZIF-8) nanobubbles with a uniform size of less than 100 nm. Interestingly, the ZIF-8 nanobubbles possess a monocrystalline nanoshell with a thickness of around 10 nm. Under optimal pyrolytic conditions, the ZIF-8 nanobubbles can be converted into hollow carbon nanobubbles while keeping their original shapes. The structure of the nanobubble enhances the fast Na(+)/K(+) ion intercalation performance. Such remarkable improvement cannot be realized by conventional MOFs or their derived carbons.

  7. Silica substrate or portion formed from oxidation of monocrystalline silicon

    DOEpatents

    Matzke, Carolyn M.; Rieger, Dennis J.; Ellis, Robert V.

    2003-07-15

    A method is disclosed for forming an inclusion-free silica substrate using a monocrystalline silicon substrate as the starting material and oxidizing the silicon substrate to convert it entirely to silica. The oxidation process is performed from both major surfaces of the silicon substrate using a conventional high-pressure oxidation system. The resulting product is an amorphous silica substrate which is expected to have superior etching characteristics for microfabrication than conventional fused silica substrates. The present invention can also be used to convert only a portion of a monocrystalline silicon substrate to silica by masking the silicon substrate and locally thinning a portion the silicon substrate prior to converting the silicon portion entirely to silica. In this case, the silica formed by oxidizing the thinned portion of the silicon substrate can be used, for example, as a window to provide optical access through the silicon substrate.

  8. Sunlight-thin nanophotonic monocrystalline silicon solar cells

    NASA Astrophysics Data System (ADS)

    Depauw, Valérie; Trompoukis, Christos; Massiot, Inès; Chen, Wanghua; Dmitriev, Alexandre; Cabarrocas, Pere Roca i.; Gordon, Ivan; Poortmans, Jef

    2017-09-01

    Introducing nanophotonics into photovoltaics sets the path for scaling down the surface texture of crystalline-silicon solar cells from the micro- to the nanoscale, allowing to further boost the photon absorption while reducing silicon material loss. However, keeping excellent electrical performance has proven to be very challenging, as the absorber is damaged by the nanotexturing and the sensitivity to the surface recombination is dramatically increased. Here we realize a light-wavelength-scale nanotextured monocrystalline silicon cell with the confirmed efficiency of 8.6% and an effective thickness of only 830 nm. For this we adopt a self-assembled large-area and industry-compatible amorphous ordered nanopatterning, combined with an advanced surface passivation, earning strongly enhanced solar light absorption while retaining efficient electron collection. This prompts the development of highly efficient flexible and semitransparent photovoltaics, based on the industrially mature monocrystalline silicon technology.

  9. Method and apparatus for drawing monocrystalline ribbon from a melt

    DOEpatents

    Ciszek, Theodore F.; Schwuttke, Guenter H.

    1981-11-10

    A method and apparatus for drawing a monocrystalline ribbon or web from a melt comprising utilizing a shaping die including at least two elements spaced one from the other each having a portion thereof located below the level of the melt and another portion located above the level of the melt a distance sufficient to form a raised meniscus of melt about the corresponding element.

  10. Nanostructured porous Si-based nanoparticles for targeted drug delivery

    PubMed Central

    Shahbazi, Mohammad-Ali; Herranz, Barbara; Santos, Hélder A.

    2012-01-01

    One of the backbones in nanomedicine is to deliver drugs specifically to unhealthy cells. Drug nanocarriers can cross physiological barriers and access different tissues, which after proper surface biofunctionalization can enhance cell specificity for cancer therapy. Recent developments have highlighted the potential of mesoporous silica (PSiO2) and silicon (PSi) nanoparticles for targeted drug delivery. In this review, we outline and discuss the most recent advances on the applications and developments of cancer therapies by means of PSiO2 and PSi nanomaterials. Bio-engineering and fine tuning of anti-cancer drug vehicles, high flexibility and potential for sophisticated release mechanisms make these nanostructures promising candidates for “smart” cancer therapies. As a result of their physicochemical properties they can be controllably loaded with large amounts of drugs and coupled to homing molecules to facilitate active targeting. The main emphasis of this review will be on the in vitro and in vivo studies. PMID:23507894

  11. Picosecond optical vortex pulse illumination forms a monocrystalline silicon needle

    PubMed Central

    Takahashi, Fuyuto; Miyamoto, Katsuhiko; Hidai, Hirofumi; Yamane, Keisaku; Morita, Ryuji; Omatsu, Takashige

    2016-01-01

    The formation of a monocrystalline silicon needle by picosecond optical vortex pulse illumination was demonstrated for the first time in this study. The dynamics of this silicon needle formation was further revealed by employing an ultrahigh-speed camera. The melted silicon was collected through picosecond pulse deposition to the dark core of the optical vortex, forming the silicon needle on a submicrosecond time scale. The needle was composed of monocrystalline silicon with the same lattice index (100) as that of the silicon substrate, and had a height of approximately 14 μm and a thickness of approximately 3 μm. Overlaid vortex pulses allowed the needle to be shaped with a height of approximately 40 μm without any changes to the crystalline properties. Such a monocrystalline silicon needle can be applied to devices in many fields, such as core–shell structures for silicon photonics and photovoltaic devices as well as nano- or microelectromechanical systems. PMID:26907639

  12. Investigation of machining mechanism of monocrystalline silicon in nanometric grinding

    NASA Astrophysics Data System (ADS)

    He, Liping; Zhu, Fulong; Liu, Yuhong; Liu, Sheng

    2017-05-01

    Monocrystalline silicon is the foundation of the computer industry, so it has a great significance to study the ultra-high precision machining of silicon. Molecular dynamics has been proved as a very effective method for the study of ultra-precision machining in nanoscale. During the grinding of brittle materials in nano-level, there are some unique phenomena such as brittle-ductile transition. To study the machining mechanism in nanometric grinding of monocrystalline silicon, the subsurface damage of < 0 0 1 > oriented Monocrystalline silicon under different grinding speeds were investigated by means of molecular dynamics simulations. The interactions between different atoms are described by the Morse and Tersoff potential. Based on analyzing the mechanism of diamond tool extrusion induced silicon lattice slip and distortion, the grinding process is explained. The movement of atoms and phase transformation are studied. The results show that there is not enough time for atoms beneath the tool to rearrange when increasing grinding speed at a low speed, which leads to the subsurface damage thickness decreases. When the diamond tool radius is small enough but still bigger than the undeformed chip thickness, the brittle-ductile transition can be achieved in the grinding region. And during the grinding process, the normal force is smaller than the tangential force. However, when the radius of the diamond tool increases to a certain value, the normal force could be larger than the tangential force.

  13. Dual-Functional Nanoparticles Targeting CXCR4 and Delivering Antiangiogenic siRNA Ameliorate Liver Fibrosis.

    PubMed

    Liu, Chun-Hung; Chan, Kun-Ming; Chiang, Tsaiyu; Liu, Jia-Yu; Chern, Guann-Gen; Hsu, Fu-Fei; Wu, Yu-Hsuan; Liu, Ya-Chi; Chen, Yunching

    2016-07-05

    The progression of liver fibrosis, an intrinsic response to chronic liver injury, is associated with hepatic hypoxia, angiogenesis, abnormal inflammation, and significant matrix deposition, leading to the development of cirrhosis and hepatocellular carcinoma (HCC). Due to the complex pathogenesis of liver fibrosis, antifibrotic drug development has faced the challenge of efficiently and specifically targeting multiple pathogenic mechanisms. Therefore, CXCR4-targeted nanoparticles (NPs) were formulated to deliver siRNAs against vascular endothelial growth factor (VEGF) into fibrotic livers to block angiogenesis during the progression of liver fibrosis. AMD3100, a CXCR4 antagonist that was incorporated into the NPs, served dual functions: it acted as a targeting moiety and suppressed the progression of fibrosis by inhibiting the proliferation and activation of hepatic stellate cells (HSCs). We demonstrated that CXCR4-targeted NPs could deliver VEGF siRNAs to fibrotic livers, decrease VEGF expression, suppress angiogenesis and normalize the distorted vessels in the fibrotic livers in the carbon tetrachloride (CCl4) induced mouse model. Moreover, blocking SDF-1α/CXCR4 by CXCR4-targeted NPs in combination with VEGF siRNA significantly prevented the progression of liver fibrosis in CCl4-treated mice. In conclusion, the multifunctional CXCR4-targeted NPs delivering VEGF siRNAs provide an effective antifibrotic therapeutic strategy.

  14. Biodegradable Inorganic Nanovector: Passive versus Active Tumor Targeting in siRNA Transportation.

    PubMed

    Park, Dae-Hwan; Cho, Jaeyong; Kwon, Oh-Joon; Yun, Chae-Ok; Choy, Jin-Ho

    2016-03-24

    The biodegradable inorganic nanovector based on a layered double hydroxide (LDH) holds great promise for gene and drug delivery systems. However, in vivo targeted delivery of genes through LDH still remains a key challenge in the development of RNA interference therapeutics. Here, we describe in vivo and in vitro delivery system for Survivin siRNA (siSurvivin) assembled with passive LDH with a particle size of 100 nm or active LDH conjugated with a cancer overexpressing receptor targeting ligand, folic acid (LDHFA), conferring them an ability to target the tumor by either EPR-based clathrin-mediated or folate receptor-mediated endocytosis. When not only transfected into KB cells but also injected into xenograft mice, LDHFA/siSurvivin induced potent gene silencing at mRNA and protein levels in vitro, and consequently achieved a 3.0-fold higher suppression of tumor volume than LDH/siSurvivin in vivo. This anti-tumor effect was attributed to a selectively 1.2-fold higher accumulation of siSurvivin in tumor tissue compared with other organs. Targeting to the tumor with inorganic nanovector can guide and accelerate an evolution of next-generation theranosis system. © 2016 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  15. Investigation of surface passivation schemes for p-type monocrystalline silicon solar cell

    NASA Astrophysics Data System (ADS)

    Rahman, Md. Momtazur; Udoy, Ariful Banna

    2016-10-01

    This paper represents an experiment to analyze the dark saturation current densities of passivated surfaces for monocrystalline silicon solar cells. The samples are diffused at peak temperatures of 800-950 °C. Basically, symmetrical lifetime samples with different doping profiles are prepared with alkaline textured and saw damage etched (planar) surfaces. After POCl3 diffusion, the phosphorous silicate glass layers are removed in a wet chemical etching step. Several designs are chosen for the determination of the sheet resistance ( R sh), the concentration profile for excess charge carrier and the minority carrier effective lifetime of the diffused surfaces. The dark saturation current densities ( J o ) and the doping profiles are determined accordingly via quasi-steady state photoconductance decay measurement and electrochemical capacitance-voltage measurement. Three different passivation schemes are investigated as follows: silicon nitride (SiN x ) deposited by plasma-enhanced chemical vapor deposition (PECVD) technique, silicon-rich oxynitride (SiriO x N y ) capped with a PECVD SiN x layer, and thin thermally grown oxide, capped with a PECVD SiN x layer.

  16. siRNA Targeting the 2Apro Genomic Region Prevents Enterovirus 71 Replication In Vitro

    PubMed Central

    Kong, Zhenzhen; Shao, Qixiang; Su, Zhaoliang; Wang, Shengjun; Chen, Jianguo

    2016-01-01

    Enterovirus 71 (EV71) is the most important etiological agent of hand, foot, and mouth disease (HFMD) in young children, which is associated with severe neurological complications and has caused significant mortalities in recent HFMD outbreaks in Asia. However, there is no effective antiviral therapy against EV71. In this study, RNA interference (RNAi) was used as an antiviral strategy to inhibit EV71 replication. Three small interfering RNAs (siRNAs) targeting the 2Apro region of the EV71 genome were designed and synthesized. All the siRNAs were transfected individually into rhabdomyosarcoma (RD) cells, which were then infected with strain EV71-2006-52-9. The cytopathic effects (CPEs) in the infected RD cells, cell viability, viral titer, and viral RNA and protein expression were examined to evaluate the specific viral inhibition by the siRNAs. The results of cytopathogenicity and MTT tests indicated that the RD cells transfected with the three siRNAs showed slight CPEs and significantly high viability. The 50% tissue culture infective dose (TCID50) values demonstrated that the viral titer of the groups treated with three siRNAs were lower than those of the control groups. qRT–PCR and western blotting revealed that the levels of viral RNA and protein in the RD cells treated with the three siRNAs were lower than those in the controls. When RD cells transfected with siRNAs were also infected with strain EV71-2008-43-16, the expression of the VP1 protein was significantly inhibited. The levels of interferon α (IFN-α) and IFN-β did not differ significantly in any group. These results suggest that siRNAs targeting the 2Apro region of the EV71 genome exerted antiviral effects in vitro. PMID:26886455

  17. Molecularly self-assembled nucleic acid nanoparticles for targeted in vivo siRNA delivery.

    PubMed

    Lee, Hyukjin; Lytton-Jean, Abigail K R; Chen, Yi; Love, Kevin T; Park, Angela I; Karagiannis, Emmanouil D; Sehgal, Alfica; Querbes, William; Zurenko, Christopher S; Jayaraman, Muthusamy; Peng, Chang G; Charisse, Klaus; Borodovsky, Anna; Manoharan, Muthiah; Donahoe, Jessica S; Truelove, Jessica; Nahrendorf, Matthias; Langer, Robert; Anderson, Daniel G

    2012-06-03

    Nanoparticles are used for delivering therapeutics into cells. However, size, shape, surface chemistry and the presentation of targeting ligands on the surface of nanoparticles can affect circulation half-life and biodistribution, cell-specific internalization, excretion, toxicity and efficacy. A variety of materials have been explored for delivering small interfering RNAs (siRNAs)--a therapeutic agent that suppresses the expression of targeted genes. However, conventional delivery nanoparticles such as liposomes and polymeric systems are heterogeneous in size, composition and surface chemistry, and this can lead to suboptimal performance, a lack of tissue specificity and potential toxicity. Here, we show that self-assembled DNA tetrahedral nanoparticles with a well-defined size can deliver siRNAs into cells and silence target genes in tumours. Monodisperse nanoparticles are prepared through the self-assembly of complementary DNA strands. Because the DNA strands are easily programmable, the size of the nanoparticles and the spatial orientation and density of cancer-targeting ligands (such as peptides and folate) on the nanoparticle surface can be controlled precisely. We show that at least three folate molecules per nanoparticle are required for optimal delivery of the siRNAs into cells and, gene silencing occurs only when the ligands are in the appropriate spatial orientation. In vivo, these nanoparticles showed a longer blood circulation time (t(1/2) ≈ 24.2 min) than the parent siRNA (t(1/2) ≈ 6 min).

  18. RNA nanoparticle as a vector for targeted siRNA delivery into glioblastoma mouse model

    PubMed Central

    Lee, Tae Jin; Haque, Farzin; Shu, Dan; Yoo, Ji Young; Li, Hui; Yokel, Robert A.; Horbinski, Craig; Kim, Tae Hyong; Kim, Sung-Hak; Kwon, Chang-Hyuk; Nakano, Ichiro; Kaur, Balveen; Guo, Peixuan; Croce, Carlo M.

    2015-01-01

    Systemic siRNA administration to target and treat glioblastoma, one of the most deadly cancers, requires robust and efficient delivery platform without immunogenicity. Here we report newly emerged multivalent naked RNA nanoparticle (RNP) based on pRNA 3-way-junction (3WJ) from bacteriophage phi29 to target glioblastoma cells with folate (FA) ligand and deliver siRNA for gene silencing. Systemically injected FA-pRNA-3WJ RNPs successfully targeted and delivered siRNA into brain tumor cells in mice, and efficiently reduced luciferase reporter gene expression (4-fold lower than control). The FA-pRNA-3WJ RNP also can target human patient-derived glioblastoma stem cells, thought to be responsible for tumor initiation and deadly recurrence, without accumulation in adjacent normal brain cells, nor other major internal organs. This study provides possible application of pRNA-3WJ RNP for specific delivery of therapeutics such as siRNA, microRNA and/or chemotherapeutic drugs into glioblastoma cells without inflicting collateral damage to healthy tissues. PMID:25885522

  19. RNA nanoparticle as a vector for targeted siRNA delivery into glioblastoma mouse model.

    PubMed

    Lee, Tae Jin; Haque, Farzin; Shu, Dan; Yoo, Ji Young; Li, Hui; Yokel, Robert A; Horbinski, Craig; Kim, Tae Hyong; Kim, Sung-Hak; Kwon, Chang-Hyuk; Nakano, Ichiro; Kaur, Balveen; Guo, Peixuan; Croce, Carlo M

    2015-06-20

    Systemic siRNA administration to target and treat glioblastoma, one of the most deadly cancers, requires robust and efficient delivery platform without immunogenicity. Here we report newly emerged multivalent naked RNA nanoparticle (RNP) based on pRNA 3-way-junction (3WJ) from bacteriophage phi29 to target glioblastoma cells with folate (FA) ligand and deliver siRNA for gene silencing. Systemically injected FA-pRNA-3WJ RNPs successfully targeted and delivered siRNA into brain tumor cells in mice, and efficiently reduced luciferase reporter gene expression (4-fold lower than control). The FA-pRNA-3WJ RNP also can target human patient-derived glioblastoma stem cells, thought to be responsible for tumor initiation and deadly recurrence, without accumulation in adjacent normal brain cells, nor other major internal organs. This study provides possible application of pRNA-3WJ RNP for specific delivery of therapeutics such as siRNA, microRNA and/or chemotherapeutic drugs into glioblastoma cells without inflicting collateral damage to healthy tissues.

  20. Molecularly Self-Assembled Nucleic Acid Nanoparticles for Targeted In Vivo siRNA Delivery

    PubMed Central

    Lee, Hyukjin; Lytton-Jean, Abigail K. R.; Chen, Yi; Love, Kevin T.; Park, Angela I.; Karagiannis, Emmanouil D.; Sehgal, Alfica; Querbes, William; Zurenko, Christopher S.; Jayaraman, Muthusamy; Peng, Chang G.; Charisse, Klaus; Borodovsky, Anna; Manoharan, Muthiah; Donahoe, Jessica S.; Truelove, Jessica; Nahrendorf, Matthias; Langer, Robert; Anderson, Daniel G.

    2013-01-01

    Nanoparticles are employed for delivering therapeutics into cells1,2. However, size, shape, surface chemistry and the presentation of targeting ligands on the surface of nanoparticles can affect circulation half-life and biodistribution, cell specific internalization, excretion, toxicity, and efficacy3-7. A variety of materials have been explored for delivering small interfering RNAs (siRNAs) - a therapeutic agent that suppresses the expression of targeted genes8,9. However, conventional delivery nanoparticles such as liposomes and polymeric systems are heterogeneous in size, composition and surface chemistry, and this can lead to suboptimal performance, lack of tissue specificity and potential toxicity10-12. Here, we show that self-assembled DNA tetrahedral nanoparticles with a well-defined size can deliver siRNAs into cells and silence target genes in tumours. Monodisperse nanoparticles are prepared through the self-assembly of complementary DNA strands. Because the DNA strands are easily programmable, the size of the nanoparticles and the spatial orientation and density of cancer targeting ligands (such as peptides and folate) on the nanoparticle surface can be precisely controlled. We show that at least three folate molecules per nanoparticle is required for optimal delivery of the siRNAs into cells and, gene silencing occurs only when the ligands are in the appropriate spatial orientation. In vivo, these nanoparticles showed a longer blood circulation time (t1/2 ∼ 24.2 min) than the parent siRNA (t1/2 ∼ 6 min). PMID:22659608

  1. Molecularly self-assembled nucleic acid nanoparticles for targeted in vivo siRNA delivery

    NASA Astrophysics Data System (ADS)

    Lee, Hyukjin; Lytton-Jean, Abigail K. R.; Chen, Yi; Love, Kevin T.; Park, Angela I.; Karagiannis, Emmanouil D.; Sehgal, Alfica; Querbes, William; Zurenko, Christopher S.; Jayaraman, Muthusamy; Peng, Chang G.; Charisse, Klaus; Borodovsky, Anna; Manoharan, Muthiah; Donahoe, Jessica S.; Truelove, Jessica; Nahrendorf, Matthias; Langer, Robert; Anderson, Daniel G.

    2012-06-01

    Nanoparticles are used for delivering therapeutics into cells. However, size, shape, surface chemistry and the presentation of targeting ligands on the surface of nanoparticles can affect circulation half-life and biodistribution, cell-specific internalization, excretion, toxicity and efficacy. A variety of materials have been explored for delivering small interfering RNAs (siRNAs)--a therapeutic agent that suppresses the expression of targeted genes. However, conventional delivery nanoparticles such as liposomes and polymeric systems are heterogeneous in size, composition and surface chemistry, and this can lead to suboptimal performance, a lack of tissue specificity and potential toxicity. Here, we show that self-assembled DNA tetrahedral nanoparticles with a well-defined size can deliver siRNAs into cells and silence target genes in tumours. Monodisperse nanoparticles are prepared through the self-assembly of complementary DNA strands. Because the DNA strands are easily programmable, the size of the nanoparticles and the spatial orientation and density of cancer-targeting ligands (such as peptides and folate) on the nanoparticle surface can be controlled precisely. We show that at least three folate molecules per nanoparticle are required for optimal delivery of the siRNAs into cells and, gene silencing occurs only when the ligands are in the appropriate spatial orientation. In vivo, these nanoparticles showed a longer blood circulation time (t1/2 ~ 24.2 min) than the parent siRNA (t1/2 ~ 6 min).

  2. Fluorescence imaging as a diagnostic of M-band x-ray drive condition in hohlraum with fluorescent Si targets

    NASA Astrophysics Data System (ADS)

    Li, Qi; Hu, Zhimin; Yao, Li; Huang, Chengwu; Yuan, Zheng; Zhao, Yang; Xiong, Gang; Qing, Bo; Lv, Min; Zhu, Tuo; Deng, Bo; Li, Jin; Wei, Minxi; Zhan, Xiayu; Li, Jun; Yang, Yimeng; Su, Chunxiao; Yang, Guohong; Zhang, Jiyan; Li, Sanwei; Yang, Jiamin; Ding, Yongkun

    2017-01-01

    Fluorescence imaging of surrogate Si-doped CH targets has been used to provide a measurement for drive condition of high-energy x-ray (i.e. M-band x-ray) drive symmetry upon the capsule in hohlraum on Shenguang-II laser facility. A series of experiments dedicated to the study of photo-pumping and fluorescence effect in Si-plasma are presented. To investigate the feasibility of fluorescence imaging in Si-plasma, an silicon plasma in Si-foil target is pre-formed at ground state by the soft x-ray from a half-hohlraum, which is then photo-pumped by the K-shell lines from a spatially distinct laser-produced Si-plasma. The resonant Si photon pump is used to improve the fluorescence signal and cause visible image in the Si-foil. Preliminary fluorescence imaging of Si-ball target is performed in both Si-doped and pure Au hohlraum. The usual capsule at the center of the hohlraum is replaced with a solid Si-doped CH-ball (Si-ball). Since the fluorescence is proportional to the photon pump upon the Si-plasma, high-energy x-ray drive symmetry is equal to the fluorescence distribution of the Si-ball.

  3. Recombinant high density lipoprotein nanoparticles for target-specific delivery of siRNA.

    PubMed

    Rui, Mengjie; Tang, Hailing; Li, Yan; Wei, Xiaohui; Xu, Yuhong

    2013-05-01

    Regulation of gene expression using small interfering RNA (siRNA) is a promising strategy for treatments of numerous diseases. However, the progress towards broad application of siRNA requires the development of safe and effective vectors that target to specific cells. In this study, we developed a novel recombinant high density lipoprotein (rHDL) vector with high siRNA encapsulation efficiency. They were prepared by condensing siRNA with various commercial cationic polymers and coating the polyplex with a layer of lipids and apolipoprotein AI (apo AI). The rHDL nanoparticles were used to transfect SMMC-7721 hepatoma cells with stable luciferase expression. The uptake and intracellular trafficing of siRNA were also investigated. Characterization studies revealed these rHDL nanoparticles had similar physical properties as natural HDLs. The various rHDL formulations had high silencing efficiency (more than 70% knockdown) in hepatocytes with minimum cytotoxicity. Moreover, the uptake of rHDL by SMMC-7721 was confirmed to be mediated through the natural HDL uptake pathway. The work described here demonstrated the optimized rHDL nanoparticles may offer a promising tool for siRNA delivery to the liver.

  4. Targeted delivery of siRNA to macrophages for anti-inflammatory treatment.

    PubMed

    Kim, Sang-Soo; Ye, Chunting; Kumar, Priti; Chiu, Isaac; Subramanya, Sandesh; Wu, Haoquan; Shankar, Premlata; Manjunath, N

    2010-05-01

    Inflammation mediated by tumor necrosis factor-alpha (TNF-alpha) and the associated neuronal apoptosis characterizes a number of neurologic disorders. Macrophages and microglial cells are believed to be the major source of TNF-alpha in the central nervous system (CNS). Here, we show that suppression of TNF-alpha by targeted delivery of small interfering RNA (siRNA) to macrophage/microglial cells dramatically reduces lipopolysaccharide (LPS)-induced neuroinflammation and neuronal apoptosis in vivo. Because macrophage/microglia express the nicotinic acetylcholine receptor (AchR) on their surface, we used a short AchR-binding peptide derived from the rabies virus glycoprotein (RVG) as a targeting ligand. This peptide was fused to nona-D-arginine residues (RVG-9dR) to enable siRNA binding. RVG-9dR was able to deliver siRNA to induce gene silencing in macrophages and microglia cells from wild type, but not AchR-deficient mice, confirming targeting specificity. Treatment with anti-TNF-alpha siRNA complexed to RVG-9dR achieved efficient silencing of LPS-induced TNF-alpha production by primary macrophages and microglia cells in vitro. Moreover, intravenous injection with RVG-9dR-complexed siRNA in mice reduced the LPS-induced TNF-alpha levels in blood as well as in the brain, leading to a significant reduction in neuronal apoptosis. These results demonstrate that RVG-9dR provides a tool for siRNA delivery to macrophages and microglia and that suppression of TNF-alpha can potentially be used to suppress neuroinflammation in vivo.

  5. Dynamic PolyConjugates for targeted in vivo delivery of siRNA to hepatocytes.

    PubMed

    Rozema, David B; Lewis, David L; Wakefield, Darren H; Wong, So C; Klein, Jason J; Roesch, Paula L; Bertin, Stephanie L; Reppen, Tom W; Chu, Qili; Blokhin, Andrei V; Hagstrom, James E; Wolff, Jon A

    2007-08-07

    Achieving efficient in vivo delivery of siRNA to the appropriate target cell would be a major advance in the use of RNAi in gene function studies and as a therapeutic modality. Hepatocytes, the key parenchymal cells of the liver, are a particularly attractive target cell type for siRNA delivery given their central role in several infectious and metabolic disorders. We have developed a vehicle for the delivery of siRNA to hepatocytes both in vitro and in vivo, which we have named siRNA Dynamic PolyConjugates. Key features of the Dynamic PolyConjugate technology include a membrane-active polymer, the ability to reversibly mask the activity of this polymer until it reaches the acidic environment of endosomes, and the ability to target this modified polymer and its siRNA cargo specifically to hepatocytes in vivo after simple, low-pressure i.v. injection. Using this delivery technology, we demonstrate effective knockdown of two endogenous genes in mouse liver: apolipoprotein B (apoB) and peroxisome proliferator-activated receptor alpha (ppara). Knockdown of apoB resulted in clear phenotypic changes that included a significant reduction in serum cholesterol and increased fat accumulation in the liver, consistent with the known functions of apoB. Knockdown of ppara also resulted in a phenotype consistent with its known function, although with less penetrance than observed in apoB knockdown mice. Analyses of serum liver enzyme and cytokine levels in treated mice indicated that the siRNA Dynamic PolyConjugate was nontoxic and well tolerated.

  6. Dynamic PolyConjugates for targeted in vivo delivery of siRNA to hepatocytes

    PubMed Central

    Rozema, David B.; Lewis, David L.; Wakefield, Darren H.; Wong, So C.; Klein, Jason J.; Roesch, Paula L.; Bertin, Stephanie L.; Reppen, Tom W.; Chu, Qili; Blokhin, Andrei V.; Hagstrom, James E.; Wolff, Jon A.

    2007-01-01

    Achieving efficient in vivo delivery of siRNA to the appropriate target cell would be a major advance in the use of RNAi in gene function studies and as a therapeutic modality. Hepatocytes, the key parenchymal cells of the liver, are a particularly attractive target cell type for siRNA delivery given their central role in several infectious and metabolic disorders. We have developed a vehicle for the delivery of siRNA to hepatocytes both in vitro and in vivo, which we have named siRNA Dynamic PolyConjugates. Key features of the Dynamic PolyConjugate technology include a membrane-active polymer, the ability to reversibly mask the activity of this polymer until it reaches the acidic environment of endosomes, and the ability to target this modified polymer and its siRNA cargo specifically to hepatocytes in vivo after simple, low-pressure i.v. injection. Using this delivery technology, we demonstrate effective knockdown of two endogenous genes in mouse liver: apolipoprotein B (apoB) and peroxisome proliferator-activated receptor alpha (ppara). Knockdown of apoB resulted in clear phenotypic changes that included a significant reduction in serum cholesterol and increased fat accumulation in the liver, consistent with the known functions of apoB. Knockdown of ppara also resulted in a phenotype consistent with its known function, although with less penetrance than observed in apoB knockdown mice. Analyses of serum liver enzyme and cytokine levels in treated mice indicated that the siRNA Dynamic PolyConjugate was nontoxic and well tolerated. PMID:17652171

  7. Novel targeted therapy for neuroblastoma: silencing the MXD3 gene using siRNA.

    PubMed

    Duong, Connie; Yoshida, Sakiko; Chen, Cathy; Barisone, Gustavo; Diaz, Elva; Li, Yueju; Beckett, Laurel; Chung, Jong; Antony, Reuben; Nolta, Jan; Nitin, Nitin; Satake, Noriko

    2017-09-01

    BackgroundNeuroblastoma is the second most common extracranial cancer in children. Current therapies for neuroblastoma, which use a combination of chemotherapy drugs, have limitations for high-risk subtypes and can cause significant long-term adverse effects in young patients. Therefore, a new therapy is needed. In this study, we investigated the transcription factor MXD3 as a potential therapeutic target in neuroblastoma.MethodsMXD3 expression was analyzed in five neuroblastoma cell lines by immunocytochemistry and quantitative real-time reverse transcription PCR, and in 18 primary patient tumor samples by immunohistochemistry. We developed nanocomplexes using siRNA and superparamagnetic iron oxide nanoparticles to target MXD3 in neuroblastoma cell lines in vitro as a single-agent therapeutic and in combination with doxorubicin, vincristine, cisplatin, or maphosphamide-common drugs used in current neuroblastoma treatment.ResultsMXD3 was highly expressed in neuroblastoma cell lines and in patient tumors that had high-risk features. Neuroblastoma cells treated in vitro with the MXD3 siRNA nanocomplexes showed MXD3 protein knockdown and resulted in cell apoptosis. Furthermore, on combining MXD3 siRNA nanocomplexes with each of the four drugs, all showed additive efficacy.ConclusionThese results indicate that MXD3 is a potential new target and that the use of MXD3 siRNA nanocomplexes is a novel therapeutic approach for neuroblastoma.

  8. Phosphorus and boron diffusion gettering of iron in monocrystalline silicon

    NASA Astrophysics Data System (ADS)

    Talvitie, H.; Vähänissi, V.; Haarahiltunen, A.; Yli-Koski, M.; Savin, H.

    2011-05-01

    We have studied experimentally the phosphorus diffusion gettering (PDG) of iron in monocrystalline silicon at the temperature range of 650-800 °C. Our results fill the lack of data at low temperatures so that we can obtain a reliable segregation coefficient for iron between a phosphorus diffused layer and bulk silicon. The improved segregation coefficient is verified by time dependent PDG simulations. Comparison of the PDG to boron diffusion gettering (BDG) in the same temperature range shows PDG to be only slightly more effective than BDG. In general, we found that BDG requires more carefully designed processing conditions than PDG to reach a high gettering efficiency.

  9. Non-targeting siRNA induces NPGPx expression to cooperate with exoribonuclease XRN2 for releasing the stress.

    PubMed

    Wei, Pei-Chi; Lo, Wen-Ting; Su, Mei-I; Shew, Jin-Yuh; Lee, Wen-Hwa

    2012-01-01

    Short interfering RNAs (siRNAs) target specific mRNAs for their degradation mediated by RNA-induced silencing complex (RISC). Persistent activation of siRNA-RISC frequently leads to non-targeting toxicity. However, how cells mediate this stress remains elusive. In this communication, we found that the presence of non-targeting siRNA selectively induced the expression of an endoplasmic reticulum (ER)-resident protein, non-selenocysteine containing phospholipid hydroperoxide glutathione peroxidase (NPGPx), but not other ER-stress proteins including GRP78, Calnexin and XBP1. Cells suffering from constant non-targeting siRNA stress grew slower and prolonged G1 phase, while NPGPx-depleted cells accumulated mature non-targeting siRNA and underwent apoptosis. Upon the stress, NPGPx covalently bound to exoribonuclease XRN2, facilitating XRN2 to remove accumulated non-targeting siRNA. These results suggest that NPGPx serves as a novel responder to non-targeting siRNA-induced stress in facilitating XRN2 to release the non-targeting siRNA accumulation.

  10. pH-responsive hybrid quantum dots for targeting hypoxic tumor siRNA delivery.

    PubMed

    Zhu, HongYan; Zhang, ShengYu; Ling, Yong; Meng, GuoLiang; Yang, Yu; Zhang, Wei

    2015-12-28

    Hypoxia is a characteristic of cancer and plays a key role in tumorigenesis, angiogenesis and resistance to cancer therapies. SiRNA treatment is effective against hypoxic tumors by gene silencing. However, siRNA delivery to the hypoxic regions of solid tumors still presents a challenge due to the distance from blood vessels and the increased presence of efflux transporters. Therefore, tumor therapies would be improved through the immediate development of an effective siRNA delivery system to hypoxic regions. To this end, we synthesized a system to deliver HIF-1α siRNA into hypoxic tumor cells. The system consists of a functional shell composed of 2-deoxyglucose (DG)-polyethylene glycol (PEG) connected with the compound of lipoic acid, lysine and 9-poly-d-arginine (LA-Lys-9R) by a hydrazone bond and a core of CdTe quantum dots (QDs). The molecular structure of DG-PEG-LA-Lys-9R was confirmed by liquid chromatography-mass spectrometry (LC-MS), nuclear magnetic resonance (NMR) spectroscopy, Fourier transform infrared spectroscopy (FTIR), and sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE). The multifunctional CdTe QDs measured approximately 200 nm and showed excellent biocompatibility, perfect siRNA binding capability and enhanced hypoxic tumor targeting. Importantly, the system described here is pH-responsive with a hydrazone bond; therefore, it avoids GLUT1 receptor-mediated endocytic recycling, resulting in irreversible delivery of the siRNA. We used Western blots to confirm the superior gene silencing efficiency induced by the DG-PEG-LA-Lys-9R with hydrazone modified CdTe QDs. Here, we demonstrate high efficacy of the siRNA tumor delivery system using in vitro and in vivo experiments. In addition, these studies demonstrate that pH-responsive hybrid quantum dots show improved antitumor efficacy with decreased organ toxicity, indicating a promising siRNA delivery system for hypoxic cancer therapy.

  11. Selective Modification of HK Peptides Enhances siRNA Silencing of Tumor Targets In Vivo

    PubMed Central

    Chou, Szu-Ting; Leng, Qixin; Scaria, Puthupparampil; Woodle, Martin; Mixson, A. James

    2011-01-01

    Our research has focused on systemic delivery of small interference RNA (siRNA) by branched peptides composed of histidine and lysine, called HK peptides. After studying several HK peptides, one four-branched peptide, H3K(+H)4b, with a predominant repeating pattern of -HHHK-, was found to be an effective carrier of siRNA. Although the unmodified H3K(+H)4b carrier of siRNA targeting an oncogene was previously shown to have promise in a tumor-bearing mouse model, we sought to develop a more effective HK carrier of siRNA in the current study. Our primary goal was to determine whether different ligand (cyclic RGD)-pegylation patterns on the H3K(+H)4b peptide affect siRNA delivery in vitro and in vivo. We compared the unmodified H3K(+H)4b with two modified H3K(+H)4b peptides for their ability to deliver siRNA in a tumor-bearing mouse model; one modified HK peptide, (RGD-PEG)4-H3K(+H)4b, had four cRGD-PEG conjugated to each molecule, while the other peptide, (RGD-PEG)-H3K(+H)4b, had one cRGD-PEG per molecule. Although the modified HK peptides by themselves did not form stable nanoplexes with siRNA, combination of a highly charged unmodified HK peptide, H2K4b, with either of the modified HK peptides did form stable siRNA nanoparticles. For in vitro experiments with MDA-MB-435 cells that expressed luciferase, the H3K(+H)4b siRNA nanoplexes targeting luciferase decreased its activity by 90% compared with negligible down-regulation by the modified H3K(+H)4b nanoplexes (P<0.01). In contrast, the two modified H3K(+H)4b siRNA nanoplexes administered intravenously were more effective than the H3K(+H)4b nanoplexes in silencing luciferase in a tumor xenograft model. The luciferase activity in tumor lysates of mice administered H3K(+H)4b, (RGD-PEG)-H3K(+H)4b, and (RGD-PEG)4-H3K(+H)4b nanoplexes decreased by 18%, 35%, and 75%, respectively. Thus, the siRNA nanoplex incorporating the highly modified peptide, (RGD-PEG)4-H3K(+H)4b, was the most effective at silencing its target in vivo

  12. Unexpected off-targeting effects of anti-huntingtin ribozymes and siRNA in vivo

    PubMed Central

    Denovan-Wright, Eileen M.; Rodriguez-Lebron, Edgardo; Lewin, Alfred S.; Mandel, Ronald J.

    2009-01-01

    Gene transfer strategies to reduce levels of mutant huntingtin (mHtt) mRNA and protein by targeting human Htt have shown therapeutic promise in vivo. Previously, we have reported that a specific, adeno-associated viral vector (rAAV)-delivered short-hairpin RNA (siHUNT-2) targeting human Htt mRNA unexpectedly decreased levels of striatal-specific transcripts in both wild-type and R6/1 transgenic HD mice. The goal of this study was to determine whether the siHUNT-2-mediated effect was due to adverse effects of RNA interference (RNAi) expression in the brain. To this end, we designed two catalytically active hammerhead ribozymes directed against the same region of human Htt mRNA targeted by siHUNT-2 and delivered them to wild-type and R6/1 transgenic HD mice. After 10 weeks of continuous expression, these ribozymes, like siHUNT-2, negatively impacted the expression of a subset of genes in the striatum. This effect was independent of rAAV transduction and specific to the targeting of a unique sequence in human Htt mRNA. After consideration of the known potential RNAi-specific toxic mechanisms, only cleavage of an unintended RNA target can account for the data reported herein. Thus, long-term rAAV-mediated RNAi in the brain does not, in and of itself, negatively affect striatal gene expression. These findings have important implications in the development of therapeutic RNAi for the treatment of neurological disease. PMID:18166484

  13. Soft computing model for optimized siRNA design by identifying off target possibilities using artificial neural network model.

    PubMed

    Murali, Reena; John, Philips George; Peter S, David

    2015-05-15

    The ability of small interfering RNA (siRNA) to do posttranscriptional gene regulation by knocking down targeted genes is an important research topic in functional genomics, biomedical research and in cancer therapeutics. Many tools had been developed to design exogenous siRNA with high experimental inhibition. Even though considerable amount of work has been done in designing exogenous siRNA, design of effective siRNA sequences is still a challenging work because the target mRNAs must be selected such that their corresponding siRNAs are likely to be efficient against that target and unlikely to accidentally silence other transcripts due to sequence similarity. In some cases, siRNAs may tolerate mismatches with the target mRNA, but knockdown of genes other than the intended target could make serious consequences. Hence to design siRNAs, two important concepts must be considered: the ability in knocking down target genes and the off target possibility on any nontarget genes. So before doing gene silencing by siRNAs, it is essential to analyze their off target effects in addition to their inhibition efficacy against a particular target. Only a few methods have been developed by considering both efficacy and off target possibility of siRNA against a gene. In this paper we present a new design of neural network model with whole stacking energy (ΔG) that enables to identify the efficacy and off target effect of siRNAs against target genes. The tool lists all siRNAs against a particular target with their inhibition efficacy and number of matches or sequence similarity with other genes in the database. We could achieve an excellent performance of Pearson Correlation Coefficient (R=0. 74) and Area Under Curve (AUC=0.906) when the threshold of whole stacking energy is ≥-34.6 kcal/mol. To the best of the author's knowledge, this is one of the best score while considering the "combined efficacy and off target possibility" of siRNA for silencing a gene. The proposed model

  14. Mannose-conjugated layered double hydroxide nanocomposite for targeted siRNA delivery to enhance cancer therapy.

    PubMed

    Li, Li; Zhang, Run; Gu, Wenyi; Xu, Zhi Ping

    2017-06-22

    Sheet-like layered double hydroxide nanoparticles (LDH NPs) have showed great potentials in biomedical applications such as nanocarriers for drug and gene delivery, biosensors and imaging agents. However, target delivery of drugs and genes using LDH NPs to the desired tumor sites is a major challenge in cancer therapy. The aim of this study is to develop a functional LDH-based nanocomposite for target delivery of siRNA to cancer cells. Mannose as a targeting moiety was firstly conjugated onto SiO2-coated LDH nanocomposite. Cellular uptake data have demonstrated that siRNA is more efficiently delivered to osteosarcoma (U2OS) cells by mannose-conjugated SiO2 coated LDH nanocomposite (Man-SiO2@LDH) compared to unmodified LDH NP. A commercial cell death-siRNA (CD-siRNA) delivered by Man-SiO2@LDH can kill cancer cells more effectively. These results reveal that the Man-SiO2@LDH nanocomposite is capable of target-delivering siRNA or drugs to tumor cells for more effective cancer treatment, which provides great potentials in cancer therapy. Copyright © 2017 Elsevier Inc. All rights reserved.

  15. High frequency guided wave propagation in monocrystalline silicon wafers

    NASA Astrophysics Data System (ADS)

    Pizzolato, Marco; Masserey, Bernard; Robyr, Jean-Luc; Fromme, Paul

    2017-04-01

    Monocrystalline silicon wafers are widely used in the photovoltaic industry for solar panels with high conversion efficiency. The cutting process can introduce micro-cracks in the thin wafers and lead to varying thickness. High frequency guided ultrasonic waves are considered for the structural monitoring of the wafers. The anisotropy of the monocrystalline silicon leads to variations of the wave characteristics, depending on the propagation direction relative to the crystal orientation. Full three-dimensional Finite Element simulations of the guided wave propagation were conducted to visualize and quantify these effects for a line source. The phase velocity (slowness) and skew angle of the two fundamental Lamb wave modes (first anti-symmetric mode A0 and first symmetric mode S0) for varying propagation directions relative to the crystal orientation were measured experimentally. Selective mode excitation was achieved using a contact piezoelectric transducer with a custom-made wedge and holder to achieve a controlled contact pressure. The out-of-plane component of the guided wave propagation was measured using a noncontact laser interferometer. Good agreement was found with the simulation results and theoretical predictions based on nominal material properties of the silicon wafer.

  16. Mechanical properties of monocrystalline and polycrystalline monolayer black phosphorus

    NASA Astrophysics Data System (ADS)

    Cao, Pinqiang; Wu, Jianyang; Zhang, Zhisen; Ning, Fulong

    2017-01-01

    The mechanical properties of monocrystalline and polycrystalline monolayer black phosphorus (MBP) are systematically investigated using classic molecular dynamic simulations. For monocrystalline MBP, it is found that the shear strain rate, sample dimensions, temperature, atomic vacancies and applied statistical ensemble affect the shear behaviour. The wrinkled morphology is closely connected with the direction of the in-plane shear, dimensions of the samples, and applied ensembles. Particularly, small samples subjected to loading/unloading of the shear deformation along the armchair direction demonstrate a clear mechanical hysteresis loop. For polycrystalline MBP, the maximum shear stress as a function of the average grain size follows an inverse pseudo Hall-Petch type relationship under an isothermal-isobaric (NPT) ensemble, whereas under a canonical (NVT) ensemble, the maximum shear stress of polycrystalline MBP exhibits a ‘flipped’ behaviour. Furthermore, polycrystalline MBP subjected to uniaxial tension also exhibits a strongly grain size-dependent mechanical response, and it can fail by brittle intergranular and transgranular fractures because of its weaker grain boundary structures and the direction-dependent edge energy, respectively. These findings provide useful insight into the mechanical design of BP for nanoelectronic devices.

  17. Monocrystalline CdTe solar cells with open-circuit voltage over 1 V and efficiency of 17%

    NASA Astrophysics Data System (ADS)

    Zhao, Yuan; Boccard, Mathieu; Liu, Shi; Becker, Jacob; Zhao, Xin-Hao; Campbell, Calli M.; Suarez, Ernesto; Lassise, Maxwell B.; Holman, Zachary; Zhang, Yong-Hang

    2016-06-01

    The open-circuit voltages of mature single-junction photovoltaic devices are lower than the bandgap energy of the absorber, typically by a gap of 400 mV. For CdTe, which has a bandgap of 1.5 eV, the gap is larger; for polycrystalline samples, the open-circuit voltage of solar cells with the record efficiency is below 900 mV, whereas for monocrystalline samples it has only recently achieved values barely above 1 V. Here, we report a monocrystalline CdTe/MgCdTe double-heterostructure solar cell with open-circuit voltages of up to 1.096 V. The latticed-matched MgCdTe barrier layers provide excellent passivation to the CdTe absorber, resulting in a carrier lifetime of 3.6 μs. The solar cells are made of 1- to 1.5-μm-thick n-type CdTe absorbers, and passivated hole-selective p-type a-SiCy:H contacts. This design allows CdTe solar cells to be made thinner and more efficient. The best power conversion efficiency achieved in a device with this structure is 17.0%.

  18. Engineering RNA for Targeted siRNA Delivery and Medical Application

    PubMed Central

    Guo, Peixuan; Coban, Oana; Snead, Nick; Trebley, Joe; Hoeprich, Steve; Guo, Songchuan; Shu, Yi

    2010-01-01

    RNA engineering for nanotechnology and medical applications is an exciting emerging research field. RNA has intrinsically defined features on the nanometer scale and is a particularly interesting candidate for such applications due to its amazing diversity, flexibility and versatility in structure and function. Specifically, the current use of siRNA to silence target genes involved in disease has generated much excitement in the scientific community. The intrinsic ability to sequence-specifically down-regulate gene expression in a temporally- and spatially-controlled fashion has led to heightened interest and rapid development of siRNA-based therapeutics. Though methods for gene silencing with high efficacy and specificity have been achieved in vitro, the effective delivery of nucleic acids to specific cells in vivo has been a hurdle for RNA therapeutics. This review covers different RNA-based approaches for diagnosis, prevention and treatment of human disease, with a focus on the latest developments of nonviral carriers of siRNA for delivery in vivo. The applications and challenges of siRNA therapy, as well as potential solutions to these problems, the approaches for using phi29 pRNA-based vectors as polyvalent vehicles for specific delivery of siRNA, ribozymes, drugs or other therapeutic agents to specific cells for therapy will also be addressed. PMID:20230868

  19. Integrated hollow mesoporous silica nanoparticles for target drug/siRNA co-delivery.

    PubMed

    Ma, Xing; Zhao, Yun; Ng, Kee Woei; Zhao, Yanli

    2013-11-11

    A hollow mesoporous silica nanoparticle (HMSNP) based drug/siRNA co-delivery system was designed and fabricated, aiming at overcoming multidrug resistance (MDR) in cancer cells for targeted cancer therapy. The as-prepared HMSNPs have perpendicular nanochannels connecting to the internal hollow cores, thereby facilitating drug loading and release. The extra volume of the hollow core enhances the drug loading capacity by two folds as compared with conventional mesoporous silica nanoparticles (MSNPs). Folic acid conjugated polyethyleneimine (PEI-FA) was coated on the HMSNP surfaces under neutral conditions through electrostatic interactions between the partially charged amino groups of PEI-FA and the phosphate groups on the HMSNP surfaces, blocking the mesopores and preventing the loaded drugs from leakage. Folic acid acts as the targeting ligand that enables the co-delivery system to selectively bind with and enter into the target cancer cells. PEI-FA-coated HMSNPs show enhanced siRNA binding capability on account of electrostatic interactions between the amino groups of PEI-FA and siRNA, as compared with that of MSNPs. The electrostatic interactions provide the feasibility of pH-controlled release. In vitro pH-responsive drug/siRNA co-delivery experiments were conducted on HeLa cell lines with high folic acid receptor expression and MCF-7 cell lines with low folic acid receptor expression for comparison, showing effective target delivery to the HeLa cells through folic acid receptor meditated cellular endocytosis. The pH-responsive intracellular drug/siRNA release greatly minimizes the prerelease and possible side effects of the delivery system. By simultaneously delivering both doxorubicin (Dox) and siRNA against the Bcl-2 protein into the HeLa cells, the expression of the anti-apoptotic protein Bcl-2 was successfully suppressed, leading to an enhanced therapeutic efficacy. Thus, the present multifunctional nanoparticles show promising potentials for controlled and

  20. Galactosylated polyaspartamide copolymers for siRNA targeted delivery to hepatocellular carcinoma cells.

    PubMed

    Cavallaro, Gennara; Farra, Rossella; Craparo, Emanuela Fabiola; Sardo, Carla; Porsio, Barbara; Giammona, Gaetano; Perrone, Francesca; Grassi, Mario; Pozzato, Gabriele; Grassi, Gabriele; Dapas, Barbara

    2017-06-20

    The limited efficacy of available treatments for hepatocellular carcinoma (HCC) requires the development of novel therapeutic approaches. We synthesized a novel cationic polymer based on α,β-poly-(N-2-hydroxyethyl)-d,L-aspartamide (PHEA) for drug delivery to HCC cells. The copolymer was synthesized by subsequent derivatization of PHEA with diethylene triamine (DETA) and with a polyethylene glycol (PEG) derivative bearing galactose (GAL) molecules, obtaining the cationic derivative PHEA-DETA-PEG-GAL. PHEA-DETA-PEG-GAL has suitable chemical-physical characteristics for a potential systemic use and can effectively deliver a siRNA (siE2F1) targeted against the transcription factor E2F1, a gene product involved in HCC. The presence of GAL residues in the polyplexes allows the targeting of HCC cells that express the asialo-glycoprotein receptor (ASGP-R). In these cells, but not in ASGP-R non-expressing cells, PHEA-DETA-PEG-GAL/siE2F1 polyplexes induce the reduction of the mRNA and protein levels of E2F1 and of E2F1-regulated genes, all involved in the promotion of the G1/S phase transition. This results in a decrease of cell proliferation with a G1/G0 phase cells accumulation. Notably, removal of GAL residue almost completely abrogates the targeting capacity of the developed polyplexes. In conclusion, the generated polyplexes demonstrate the potential to effectively contributing to the development of novel anti-HCC therapeutic approaches via a siRNA-targeted delivery. Copyright © 2017 Elsevier B.V. All rights reserved.

  1. Inter-molecular β-sheet structure facilitates lung-targeting siRNA delivery

    PubMed Central

    Zhou, Jihan; Li, Dong; Wen, Hao; Zheng, Shuquan; Su, Cuicui; Yi, Fan; Wang, Jue; Liang, Zicai; Tang, Tao; Zhou, Demin; Zhang, Li-He; Liang, Dehai; Du, Quan

    2016-01-01

    Size-dependent passive targeting based on the characteristics of tissues is a basic mechanism of drug delivery. While the nanometer-sized particles are efficiently captured by the liver and spleen, the micron-sized particles are most likely entrapped within the lung owing to its unique capillary structure and physiological features. To exploit this property in lung-targeting siRNA delivery, we designed and studied a multi-domain peptide named K-β, which was able to form inter-molecular β-sheet structures. Results showed that K-β peptides and siRNAs formed stable complex particles of 60 nm when mixed together. A critical property of such particles was that, after being intravenously injected into mice, they further associated into loose and micron-sized aggregates, and thus effectively entrapped within the capillaries of the lung, leading to a passive accumulation and gene-silencing. The large size aggregates can dissociate or break down by the shear stress generated by blood flow, alleviating the pulmonary embolism. Besides the lung, siRNA enrichment and targeted gene silencing were also observed in the liver. This drug delivery strategy, together with the low toxicity, biodegradability, and programmability of peptide carriers, show great potentials in vivo applications. PMID:26955887

  2. Inter-molecular β-sheet structure facilitates lung-targeting siRNA delivery

    NASA Astrophysics Data System (ADS)

    Zhou, Jihan; Li, Dong; Wen, Hao; Zheng, Shuquan; Su, Cuicui; Yi, Fan; Wang, Jue; Liang, Zicai; Tang, Tao; Zhou, Demin; Zhang, Li-He; Liang, Dehai; Du, Quan

    2016-03-01

    Size-dependent passive targeting based on the characteristics of tissues is a basic mechanism of drug delivery. While the nanometer-sized particles are efficiently captured by the liver and spleen, the micron-sized particles are most likely entrapped within the lung owing to its unique capillary structure and physiological features. To exploit this property in lung-targeting siRNA delivery, we designed and studied a multi-domain peptide named K-β, which was able to form inter-molecular β-sheet structures. Results showed that K-β peptides and siRNAs formed stable complex particles of 60 nm when mixed together. A critical property of such particles was that, after being intravenously injected into mice, they further associated into loose and micron-sized aggregates, and thus effectively entrapped within the capillaries of the lung, leading to a passive accumulation and gene-silencing. The large size aggregates can dissociate or break down by the shear stress generated by blood flow, alleviating the pulmonary embolism. Besides the lung, siRNA enrichment and targeted gene silencing were also observed in the liver. This drug delivery strategy, together with the low toxicity, biodegradability, and programmability of peptide carriers, show great potentials in vivo applications.

  3. Hyaluronic acid based self-assembling nanosystems for CD44 target mediated siRNA delivery to solid tumors.

    PubMed

    Ganesh, Shanthi; Iyer, Arun K; Morrissey, David V; Amiji, Mansoor M

    2013-04-01

    Anticancer therapeutics employing RNA interference mechanism holds promising potentials for sequence-specific silencing of target genes. However targeted delivery of siRNAs to tumor tissues and cells and more importantly, their intracellular release at sites of interest still remains a major challenge that needs to be addressed before this technique could become a clinically viable option. In the current study, we have engineered and screened a series of CD44 targeting hyaluronic acid (HA) based self-assembling nanosystems for targeted siRNA delivery. The HA polymer was functionalized with lipids of varying carbon chain lengths/nitrogen content, as well as polyamines for assessing siRNA encapsulation. From the screens, several HA-derivatives were identified that could stably encapsulate/complex siRNAs and form self-assembled nanosystems, as determined by gel retardation assays and dynamic light scattering. Many HA derivatives could transfect siRNAs into cancer cells overexpressing CD44 receptors. Interestingly, blocking the CD44 receptors on the cells using free excess soluble HA prior to incubation of cy3-labeled-siRNA loaded HA nano-assemblies resulted in >90% inhibition of the receptor mediated uptake, confirming target specificity. In addition, SSB/PLK1 siRNA encapsulated in HA-PEI/PEG nanosystems demonstrated dose dependent and target specific gene knockdown in both sensitive and resistant A549 lung cancer cells overexpressing CD44 receptors. More importantly, these siRNA encapsulated nanosystems demonstrated tumor selective uptake and target specific gene knock down in vivo in solid tumors as well as in metastatic tumors. The HA based nanosystems thus portend to be promising siRNA delivery vectors for systemic targeting of CD44 overexpressing cancers including tumor initiating (stem-) cells and metastatic lesions.

  4. Targeted tumor-penetrating siRNA nanocomplexes for credentialing the ovarian cancer oncogene ID4.

    PubMed

    Ren, Yin; Cheung, Hiu Wing; von Maltzhan, Geoffrey; Agrawal, Amit; Cowley, Glenn S; Weir, Barbara A; Boehm, Jesse S; Tamayo, Pablo; Karst, Alison M; Liu, Joyce F; Hirsch, Michelle S; Mesirov, Jill P; Drapkin, Ronny; Root, David E; Lo, Justin; Fogal, Valentina; Ruoslahti, Erkki; Hahn, William C; Bhatia, Sangeeta N

    2012-08-15

    The comprehensive characterization of a large number of cancer genomes will eventually lead to a compendium of genetic alterations in specific cancers. Unfortunately, the number and complexity of identified alterations complicate endeavors to identify biologically relevant mutations critical for tumor maintenance because many of these targets are not amenable to manipulation by small molecules or antibodies. RNA interference provides a direct way to study putative cancer targets; however, specific delivery of therapeutics to the tumor parenchyma remains an intractable problem. We describe a platform for the discovery and initial validation of cancer targets, composed of a systematic effort to identify amplified and essential genes in human cancer cell lines and tumors partnered with a novel modular delivery technology. We developed a tumor-penetrating nanocomplex (TPN) that comprised small interfering RNA (siRNA) complexed with a tandem tumor-penetrating and membrane-translocating peptide, which enabled the specific delivery of siRNA deep into the tumor parenchyma. We used TPN in vivo to evaluate inhibitor of DNA binding 4 (ID4) as a novel oncogene. Treatment of ovarian tumor-bearing mice with ID4-specific TPN suppressed growth of established tumors and significantly improved survival. These observations not only credential ID4 as an oncogene in 32% of high-grade ovarian cancers but also provide a framework for the identification, validation, and understanding of potential therapeutic cancer targets.

  5. Going beyond the liver: progress and challenges of targeted delivery of siRNA therapeutics.

    PubMed

    Lorenzer, Cornelia; Dirin, Mehrdad; Winkler, Anna-Maria; Baumann, Volker; Winkler, Johannes

    2015-04-10

    Therapeutic gene silencing promises significant progress in pharmacotherapy, including considerable expansion of the druggable target space and the possibility for treating orphan diseases. Technological hurdles have complicated the efficient use of therapeutic oligonucleotides, and siRNA agents suffer particularly from insufficient pharmacokinetic properties and poor cellular uptake. Intense development and evolution of delivery systems have resulted in efficient uptake predominantly in liver tissue, in which practically all nanoparticulate and liposomal delivery systems show the highest accumulation. The most efficacious strategies include liposomes and bioconjugations with N-acetylgalactosamine. Both are in early clinical evaluation stages for treatment of liver-associated diseases. Approaches for achieving knockdown in other tissues and tumors have been proven to be more complicated. Selective targeting to tumors may be enabled through careful modulation of physical properties, such as particle size, or by taking advantage of specific targeting ligands. Significant barriers stand between sufficient accumulation in other organs, including endothelial barriers, cellular membranes, and the endosome. The brain, which is shielded by the blood-brain barrier, is of particular interest to facilitate efficient oligonucleotide therapy of neurological diseases. Transcytosis of the blood-brain barrier through receptor-specific docking is investigated to increase accumulation in the central nervous system. In this review, the current clinical status of siRNA therapeutics is summarized, as well as innovative and promising preclinical concepts employing tissue- and tumor-targeted ligands. The requirements and the respective advantages and drawbacks of bioconjugates and ligand-decorated lipid or polymeric particles are discussed.

  6. Effects of targeted nano-delivery systems combined with hTERT-siRNA and Bmi-1-siRNA on MCF-7 cells

    PubMed Central

    Liu, Lei; Li, Huixiang; Zhang, Min; Lv, Xinquan

    2015-01-01

    The aim of this study was to evaluate the efficiency of a targeted siRNA nano-delivery system to silence the expression of Bmi-1 and hTERT, and to verify the toxicity of this delivery system in MCF-7 breast cancer cells. The most effective Bmi-1 siRNA and hTERT siRNA sequences were selected using RT-PCR and Western blotting. The polyethyleneimine (PEI)/siRNA nano-condensate was synthesized using PEI and modified using an NGR peptide fragment for targeting to tumor cells. The vector morphology, particle size and zeta potential were observed using an atomic force microscope and a laser particle size analyzer. The MCF-7 breast cancer cell line was transfected with the vector, and cytotoxicity was tested by MTT assays. The transfection efficiency was evaluated by qRT-PCR and Western blotting. Changes in gene expression and apoptosis rate were measured by flow cytometry. The size of LPN carrier and the condensate particle was between 100 and 200 nm and the potentials were close to neutral. There was maximum transfection efficiency and no significant increase in toxicity at 15 pmol/L. Bmi-1 and hTERT expression decreased, but the inhibition rate increased in the hTERT siRNA group, the hTERT+Bmi-1 siRNA group and the hTERT+Bmi-1 siRNA group compared with the scrambled siRNA group and the control group. Moreover, the hTERT+Bmi-1 siRNA group had the highest level of gene silencing. The complex, composed of Lipo, PEI and siRNA, is low toxicity and efficient transfection vectors. The expression level of Bmi-1 and hTERT was decreased by the gene silencing of either Bmi-1 or hTERT, but the effects were more significant when both were silenced simultaneously. PMID:26261549

  7. Modular plasmonic nanocarriers for efficient and targeted delivery of cancer-therapeutic siRNA.

    PubMed

    Huang, Xiao; Pallaoro, Alessia; Braun, Gary B; Morales, Demosthenes P; Ogunyankin, Maria O; Zasadzinski, Joseph; Reich, Norbert O

    2014-01-01

    We have combined a versatile and powerful route to deliver nucleic acids with peptide-based cell-specific targeting. siRNA targeting the polo-like kinase gene is in clinical trials for cancer treatment, and here we deliver this RNA selectively to cancer cells displaying the neuropilin-1 epitope using gold nanoshells. Release of the siRNA from the nanoparticles results from irradiation with a pulsed near-infrared laser, which also provides efficient endosomal escape within the cell. As a result, our approach requires 10-fold less material than standard nucleic acid transduction materials and is significantly more efficient than other particle-based methods. We also describe a particle-nucleic acid design that does not rely on modified RNA, thereby making the preparation of these materials more efficient and much less expensive. These improvements, when combined with control over when and where the siRNA is released, could provide the basis for diverse cell biological studies.

  8. Calculations and First Results Obtained with a SiC Prototype of the SPES Direct Target

    SciTech Connect

    Barbui, Marina; Andrighetto, Alberto; Antonucci, C.; Biasetto, Lisa; Carturan, S.; Cervellera, F.; Cevolani, S.; Cinausero, Marco; Colombo, P.; Dainelli, A.; Di Bernardo, P.; Giacchini, Mauro; Gramegna, Fabiana; Lollo, M.; Maggioni, G.; Manzolaro, Mattia; Meneghetti, G.; Petrovich, C.; Piga, L.; Prete, Gianfranco; Re, Maurizio; Rizzi, Valentina; Stracener, Daniel W; Tonezzer, Michele; Zafiropoulos, D.; Zanonato, P.

    2008-01-01

    In the framework of the SPES project at LNL [A. Bracco, A. Pisent (Ed.), REP 181/02, LNL-INFN, 2002], the realization of a direct ISOL Target for a mid-term radioactive ion beam facility is in progress. Using a primary proton beam of energy 40 MeV and intensity 0.2 mA, a high number of fission products will be obtained in the SPES multi-foil uranium carbide target, keeping a low power density deposition in the refractory matrix [A. Andrighetto, S. Cevolani, C. Petrovich, Eur. Phys J. A 25 (2005) 41]. The exotic species produced by Uranium fission in the target are collected in the ion source after the diffusion and the effusion processes. When short lived isotopes are produced it is very important to optimize the release properties of the target. To this purpose the RIBO code (radioactive ion beam optimiser) [M. Santana Leitner, A Monte Carlo Code to Optimize the Production of Radioactive Ion Beams by the ISOL Technique, PhD. Thesis, UPC-ETSEIB/CERN] has been used in order to estimate the target release efficiency for some neutron-rich nuclei. A SiC prototype of the target was recently produced at LNL and tested at ORNL using a 42 MeV proton beam. The yield of some aluminum isotopes was measured as a function of the target temperature. Some preliminary results of the data analysis will be presented.

  9. SiASR4, the Target Gene of SiARDP from Setaria italica, Improves Abiotic Stress Adaption in Plants

    PubMed Central

    Li, Jianrui; Dong, Yang; Li, Cong; Pan, Yanlin; Yu, Jingjuan

    2017-01-01

    Drought and other types of abiotic stresses negatively affect plant growth and crop yields. The abscisic acid-, stress-, and ripening-induced (ASR) proteins play important roles in the protection of plants against abiotic stress. However, the regulatory pathway of the gene encoding this protein remains to be elucidated. In this study, the foxtail millet (Setaria italica) ASR gene, SiASR4, was cloned and characterized. SiASR4 localized to the cell nucleus, cytoplasm and cytomembrane, and the protein contained 102 amino acids, including an ABA/WDS (abscisic acid/water-deficit stress) domain, with a molecular mass of 11.5 kDa. The abundance of SiASR4 transcripts increased after treatment with ABA, NaCl, and PEG in foxtail millet seedlings. It has been reported that the S. italica ABA-responsive DRE-binding protein (SiARDP) binds to a DNA sequence with a CCGAC core and that there are five dehydration-responsive element (DRE) motifs within the SiASR4 promoter. Our analyses demonstrated that the SiARDP protein could bind to the SiASR4 promoter in vitro and in vivo. The expression of SiASR4 increased in SiARDP-overexpressing plants. SiASR4-transgenic Arabidopsis and SiASR4-overexpressing foxtail millet exhibited enhanced tolerance to drought and salt stress. Furthermore, the transcription of stress-responsive and reactive oxygen species (ROS) scavenger-associated genes was activated in SiASR4 transgenic plants. Together, these findings show that SiASR4 functions in the adaption to drought and salt stress and is regulated by SiARDP via an ABA-dependent pathway. PMID:28127300

  10. Receptor-targeted liposome-peptide-siRNA nanoparticles represent an efficient delivery system for MRTF silencing in conjunctival fibrosis

    NASA Astrophysics Data System (ADS)

    Yu-Wai-Man, Cynthia; Tagalakis, Aristides D.; Manunta, Maria D.; Hart, Stephen L.; Khaw, Peng T.

    2016-02-01

    There is increasing evidence that the Myocardin-related transcription factor/Serum response factor (MRTF/SRF) pathway plays a key role in fibroblast activation and that knocking down MRTF can lead to reduced scarring and fibrosis. Here, we have developed a receptor-targeted liposome-peptide-siRNA nanoparticle as a non-viral delivery system for MRTF-B siRNA in conjunctival fibrosis. Using 50 nM siRNA, the MRTF-B gene was efficiently silenced by 76% and 72% with LYR and LER nanoparticles, respectively. The silencing efficiency was low when non-targeting peptides or siRNA alone or liposome-siRNA alone were used. LYR and LER nanoparticles also showed higher silencing efficiency than PEGylated LYR-P and LER-P nanoparticles. The nanoparticles were not cytotoxic using different liposomes, targeting peptides, and 50 nM siRNA. Three-dimensional fibroblast-populated collagen matrices were also used as a functional assay to measure contraction in vitro, and showed that MRTF-B LYR nanoparticles completely blocked matrix contraction after a single transfection treatment. In conclusion, this is the first study to develop and show that receptor-targeted liposome-peptide-siRNA nanoparticles represent an efficient and safe non-viral siRNA delivery system that could be used to prevent fibrosis after glaucoma filtration surgery and other contractile scarring conditions in the eye.

  11. Development of RNAi technology for targeted therapy--a track of siRNA based agents to RNAi therapeutics.

    PubMed

    Zhou, Yinjian; Zhang, Chunling; Liang, Wei

    2014-11-10

    RNA interference (RNAi) was intensively studied in the past decades due to its potential in therapy of diseases. The target specificity and universal treatment spectrum endowed siRNA advantages over traditional small molecules and protein drugs. However, barriers exist in the blood circulation system and the diseased tissues blocked the actualization of RNAi effect, which raised function versatility requirements to siRNA therapeutic agents. Appropriate functionalization of siRNAs is necessary to break through these barriers and target diseased tissues in local or systemic targeted application. In this review, we summarized that barriers exist in the delivery process and popular functionalized technologies for siRNA such as chemical modification and physical encapsulation. Preclinical targeted siRNA delivery and the current status of siRNA based RNAi therapeutic agents in clinical trial were reviewed and finally the future of siRNA delivery was proposed. The valuable experience from the siRNA agent delivery study and the RNAi therapeutic agents in clinical trial paved ways for practical RNAi therapeutics to emerge early.

  12. Receptor-targeted liposome-peptide-siRNA nanoparticles represent an efficient delivery system for MRTF silencing in conjunctival fibrosis

    PubMed Central

    Yu-Wai-Man, Cynthia; Tagalakis, Aristides D.; Manunta, Maria D.; Hart, Stephen L.; Khaw, Peng T.

    2016-01-01

    There is increasing evidence that the Myocardin-related transcription factor/Serum response factor (MRTF/SRF) pathway plays a key role in fibroblast activation and that knocking down MRTF can lead to reduced scarring and fibrosis. Here, we have developed a receptor-targeted liposome-peptide-siRNA nanoparticle as a non-viral delivery system for MRTF-B siRNA in conjunctival fibrosis. Using 50 nM siRNA, the MRTF-B gene was efficiently silenced by 76% and 72% with LYR and LER nanoparticles, respectively. The silencing efficiency was low when non-targeting peptides or siRNA alone or liposome-siRNA alone were used. LYR and LER nanoparticles also showed higher silencing efficiency than PEGylated LYR-P and LER-P nanoparticles. The nanoparticles were not cytotoxic using different liposomes, targeting peptides, and 50 nM siRNA. Three-dimensional fibroblast-populated collagen matrices were also used as a functional assay to measure contraction in vitro, and showed that MRTF-B LYR nanoparticles completely blocked matrix contraction after a single transfection treatment. In conclusion, this is the first study to develop and show that receptor-targeted liposome-peptide-siRNA nanoparticles represent an efficient and safe non-viral siRNA delivery system that could be used to prevent fibrosis after glaucoma filtration surgery and other contractile scarring conditions in the eye. PMID:26905457

  13. siRNA targeting PLK-1 induces apoptosis of synoviocytes in rheumatoid arthritis

    SciTech Connect

    Wada, Makoto; Kawahito, Yutaka . E-mail: kawahity@koto.kpu-m.ac.jp; Kimura, Shinya; Kohno, Masataka; Ishino, Hidetaka; Kimura, Mizuho; Omoto, Atsushi; Yamamoto, Aihiro; Hamaguchi, Masahide; Tsubouchi, Yasunori; Tokunaga, Daisaku; Hojo, Tatsuya; Ashihara, Eishi; Maekawa, Taira; Yoshikawa, Toshikazu

    2007-06-01

    Polo-like kinase-1 (PLK-1) is a member of the PLK family and participates in the control of cell mitosis. Here, we show that immunoreactive PLK-1 is strongly expressed in synoviocytes and some infiltrative mononuclear cells in synovial tissues from patients with rheumatoid arthritis (RA), while patients with osteoarthritis and injury show little or no expression of PLK-1 in synovial tissues. Western blot analysis shows that PLK is expressed and its expression is enhanced by IL-1{beta} in RA synoviocytes. IL-1{beta} also enhanced the cell growth of RA synoviocytes. Moreover, siRNA targeted against PLK-1 significantly decreases the expression of PLK-1 of RA synoviocytes stimulated by IL-1{beta} and suppresses the proliferation of these synoviocytes through apoptosis. These findings suggest that PLK-1 plays a critical role in the proliferation of RA synoviocytes leading to bone destruction, and siRNA against PLK-1 is potentially useful for the treatment of RA.

  14. Targeting BACE1 with siRNAs ameliorates Alzheimer disease neuropathology in a transgenic model.

    PubMed

    Singer, Oded; Marr, Robert A; Rockenstein, Edward; Crews, Leslie; Coufal, Nicole G; Gage, Fred H; Verma, Inder M; Masliah, Eliezer

    2005-10-01

    In Alzheimer disease, increased beta-secretase (BACE1) activity has been associated with neurodegeneration and accumulation of amyloid precursor protein (APP) products. Thus, inactivation of BACE1 could be important in the treatment of Alzheimer disease. In this study, we found that lowering BACE1 levels using lentiviral vectors expressing siRNAs targeting BACE1 reduced amyloid production and the neurodegenerative and behavioral deficits in APP transgenic mice, a model of Alzheimer disease. Our results suggest that lentiviral vector delivery of BACE1 siRNA can specifically reduce the cleavage of APP and neurodegeneration in vivo and indicate that this approach could have potential therapeutic value for treatment of Alzheimer disease.

  15. Systemic delivery of siRNA by hyaluronan-functionalized calcium phosphate nanoparticles for tumor-targeted therapy

    NASA Astrophysics Data System (ADS)

    Qiu, Chong; Wei, Wei; Sun, Jing; Zhang, Hai-Tao; Ding, Jing-Song; Wang, Jian-Cheng; Zhang, Qiang

    2016-06-01

    In this study, hyaluronan (HA)-functionalized calcium phosphate nanoparticles (CaP-AHA/siRNA NPs) were developed for an injectable and targetable delivery of siRNA, which were prepared by coating the alendronate-hyaluronan graft polymer (AHA) around the surface of calcium phosphate-siRNA co-precipitates. The prepared CaP-AHA/siRNA NPs had a uniform spherical core-shell morphology with an approximate size of 170 nm and zeta potential of -12 mV. The coating of hydrophilic HA improved the physical stability of nanoparticles over one month due to the strong interactions between phosphonate and calcium. In vitro experiments demonstrated that the negatively charged CaP-AHA/siRNA NPs could effectively deliver EGFR-targeted siRNA into A549 cells through CD44-mediated endocytosis and significantly down-regulate the level of EGFR expression. Also, the internalized CaP-AHA/siRNA NPs exhibited a pH-responsive release of siRNA, indicating that the acidification of lysosomes probably facilitated the disassembling of nanoparticles and the resultant ions sharply increased the inner osmotic pressure and thus expedited the release of siRNA from late lysosomes into the cytoplasm. Furthermore, in vivo tumor therapy demonstrated that high accumulation of CaP-AHA/siEGFR NPs in tumor led to a significant tumor growth inhibition with a specific EGFR gene silencing effect after intravenous administration in nude mice xenografted with A549 tumor, along with a negligible body weight loss. These results suggested that the CaP-AHA/siRNA NPs could be an effective and safe systemic siRNA delivery system for a RNAi-based tumor targeted therapy strategy.In this study, hyaluronan (HA)-functionalized calcium phosphate nanoparticles (CaP-AHA/siRNA NPs) were developed for an injectable and targetable delivery of siRNA, which were prepared by coating the alendronate-hyaluronan graft polymer (AHA) around the surface of calcium phosphate-siRNA co-precipitates. The prepared CaP-AHA/siRNA NPs had a uniform

  16. Evidence of RNAi in humans from systemically administered siRNA via targeted nanoparticles

    PubMed Central

    Davis, Mark E.; Zuckerman, Jonathan E.; Choi, Chung Hang J.; Seligson, David; Tolcher, Anthony; Alabi, Christopher A.; Yen, Yun; Heidel, Jeremy D.; Ribas, Antoni

    2010-01-01

    Therapeutics that are designed to engage RNA interference (RNAi) pathways have the potential to provide new, major ways of imparting therapy to patients.1,2 Fire et al. first demonstrated that long, double stranded RNAs mediate RNAi in Caenorhabditis elegans,3 and Elbashir et al. opened the pathway to the use of RNAi for human therapy by showing that small interfering RNAs (siRNAs: ca. 21 base pair double stranded RNA) can elicit RNAi in mammalian cells without producing an interferon response.4 We are currently conducting the first-in-human Phase I clinical trial involving the systemic administration of siRNA to patients with solid cancers using a targeted, nanoparticle delivery system. Here we provide evidence of inducing an RNAi mechanism of action in a human from the delivered siRNA. Tumor biopsies from melanoma patients obtained after treatment reveal: (i) the presence of intracellularly-localized nanoparticles in amounts that correlate with dose levels of the nanoparticles administered (this is a first for systemically delivered nanoparticles of any kind), and (ii) reduction in both the specific mRNA (M2 subunit of ribonucleotide reductase (RRM2)) and the protein (RRM2) when compared to pre-dosing tissue. Most importantly, we detect the presence of an mRNA fragment that demonstrates siRNA mediated mRNA cleavage occurs specifically at the site predicted for an RNAi mechanism from a patient who received the highest dose of the nanoparticles. These data when taken in total demonstrate that siRNA administered systemically to a human can produce a specific gene inhibition (reduction in mRNA and protein) by an RNAi mechanism of action. PMID:20305636

  17. Rolling circle transcription-based polymeric siRNA nanoparticles for tumor-targeted delivery.

    PubMed

    Lee, Jae Hyeop; Ku, Sook Hee; Kim, Min Ju; Lee, So Jin; Kim, Hyun Cheol; Kim, Kwangmeyung; Kim, Sun Hwa; Kwon, Ick Chan

    2017-10-10

    RNA, one of the major biological macromolecules, has been considered as an attractive building material for bottom-up fabrication of nanostructures in the past few decades due to advancements in RNA biology, RNA chemistry and RNA nanotechnology. Most recently, an isothermal enzymatic nucleic acid amplification method termed rolling circle transcription (RCT), which achieves a large-scale synthesis of RNA nanostructures, has emerged as one of fascinating techniques for RNAi-based therapies. Herein, we proposed a newly designed RCT method for synthesis of polymeric siRNA nanoflower, referred to 'RCT and annealing-generated polymeric siRNA (RAPSI)': (1) Amplification of the antisense strand of siRNA via RCT process and (2) annealing of chimeric sense strand containing 3'-terminal DNA nucleotides that provide enzyme cleavage sites. To verify its potentials in RNAi-based cancer therapy, the newly designed RAPSI nanoflower was further complexed with glycol chitosan (GC) derivatives, and systemically delivered to PC-3 xenograft tumors. The resultant RAPSI nanoparticles exhibited the improved particle stability against polyanion competition or nuclease attack. When the RAPSI nanoparticles reached to the cytoplasmic region, active mono siRNA was liberated and significantly down-regulated the expression of target VEGF gene in PC-3 cells. Excellent tumor-homing efficacy and anti-tumor effects of the RAPSI nanoparticles were further demonstrated. Overall, the proposed RCT-based polymeric siRNA nanoflower formulation can provide a new platform technology that allows further functional modifications via an advanced annealing method for systemic cancer RNAi therapy. Copyright © 2017 Elsevier B.V. All rights reserved.

  18. Galactosylated Liposomes for Targeted Co-Delivery of Doxorubicin/Vimentin siRNA to Hepatocellular Carcinoma

    PubMed Central

    Oh, Hea Ry; Jo, Hyun-Young; Park, James S.; Kim, Dong-Eun; Cho, Je-Yoel; Kim, Pyung-Hwan; Kim, Keun-Sik

    2016-01-01

    The combination of therapeutic nucleic acids and chemotherapeutic drugs has shown great promise for cancer therapy. In this study, asialoglycoprotein receptors (ASGPR) targeting-ligand-based liposomes were tested to determine whether they can co-deliver vimentin siRNA and doxorubicin to hepatocellular carcinoma (HCC) selectively. To achieve this goal, we developed an ASGPR receptor targeted co-delivery system called gal-doxorubicin/vimentin siRNA liposome (Gal-DOX/siRNA-L). The Gal-DOX/siRNA-L was created via electrostatic interaction of galactose linked-cationic liposomal doxorubicin (Gal-DOX-L) on vimentin siRNA. Previous studies have shown that Gal-DOX/siRNA-L inhibited tumor growth by combined effect of DOX and vimentin siRNA than single delivery of either DOX or vimentin siRNA. These Gal-DOX/siRNA-Ls showed stronger affinity to human hepatocellular carcinoma cells (Huh7) than other cells (lung epithelial carcinoma, A549). These liposomes also have demonstrated that novel hepatic drug/gene delivery systems composed of cationic lipid (DMKE: O,O’-dimyristyl-N-lysyl glutamate), cholesterol, galactosylated ceramide, POPC (1-palmitoyl-2-oleoyl-sn-glycero-3-phosphocholine), and PEG2000-DSPE (distearoyl phosphatidyl ethanolamine) at 2:1:1:1:0.2 (moral ratios) can be used as an effective drug/gene carrier specifically targeting the liver in vivo. These results suggest that Gal-DOX-siRNA-L could effectively target tumor cells, enhance transfection efficacy and subsequently achieve the co-delivery of DOX and siRNA, demonstrating great potential for synergistic anti-tumor therapy. PMID:28335269

  19. Atelocollagen-mediated in vivo siRNA transfection in ovarian carcinoma is influenced by tumor site, siRNA target and administration route.

    PubMed

    Meryet-Figuière, Matthieu; Lecerf, Charlotte; Varin, Emilie; Coll, Jean-Luc; Louis, Marie-Hélène; Dutoit, Soizic; Giffard, Florence; Blanc-Fournier, Cécile; Hedir, Siham; Vigneron, Nicolas; Brotin, Emilie; Pelletier, Laurent; Josserand, Véronique; Denoyelle, Christophe; Poulain, Laurent

    2017-10-01

    Ovarian cancer is the leading cause of death from gynecological malignancies worldwide, and innate or acquired chemoresistance of ovarian cancer cells is the major cause of therapeutic failure. It has been demonstrated that the concomitant inhibition of Bcl-xL and Mcl-1 anti-apoptotic activities is able to trigger apoptosis in chemoresistant ovarian cancer cells. In this context, siRNA-mediated Bcl‑xL and Mcl-1 inhibition constitutes an appealing strategy by which to eliminate chemoresistant cancer cells. However, the safest and most efficient way to vectorize siRNAs in vivo is still under debate. In the present study, using in vivo bioluminescence imaging, we evaluated the interest of atelocollagen to vectorize siRNAs by intraperitoneal (i.p.) or intravenous (i.v.) administration in 2 xenografted ovarian cancer models (peritoneal carcinomatosis and subcutaneous tumors in nude mice). Whereas i.p. administration of atelocollagen-vectorized siRNA in the peritoneal carcinomatosis model did not induce any gene downregulation, a 70% transient downregulation of luciferase expression was achieved after i.v. injection of atelocollagen-vectorized siRNA in the subcutaneous (s.c.) model. However, the use of siRNA targeting Bcl-xL or Mcl-1 did not induce target-specific downregulation in vivo in nude mice. Our results therefore show that atelocollagen complex formulation, the administration route, tumor site and the identity of the siRNA target influence the efficiency of atelocollagen‑mediated siRNA delivery.

  20. Transparent conducting Si-codoped Al-doped ZnO thin films prepared by magnetron sputtering using Al-doped ZnO powder targets containing SiC

    SciTech Connect

    Nomoto, Jun-ichi; Miyata, Toshihiro; Minami, Tadatsugu

    2009-07-15

    Transparent conducting Al-doped ZnO (AZO) thin films codoped with Si, or Si-codoped AZO (AZO:Si), were prepared by radio-frequency magnetron sputtering using a powder mixture of ZnO, Al{sub 2}O{sub 3}, and SiC as the target; the Si content (Si/[Si+Zn] atomic ratio) was varied from 0 to 1 at. %, but the Al content (Al/[Al+Zn] atomic ratio) was held constant. To investigate the effect of carbon on the electrical properties of AZO:Si thin films prepared using the powder targets containing SiC, the authors also prepared thin films using a mixture of ZnO, Al{sub 2}O{sub 3}, and SiO{sub 2} or SiO powders as the target. They found that when AZO:Si thin films were deposited on glass substrates at about 200 degree sign C, both Al and Si doped into ZnO acted as effective donors and the atomic carbon originating from the sputtered target acted as a reducing agent. As a result, sufficient improvement was obtained in the spatial distribution of resistivity on the substrate surface in AZO:Si thin films prepared with a Si content (Si/[Si+Zn] atomic ratio) of 0.75 at. % using powder targets containing SiC. The improvement in resistivity distribution was mainly attributed to increases in both carrier concentration and Hall mobility at locations on the substrate corresponding to the target erosion region. In addition, the resistivity stability of AZO: Si thin films exposed to air for 30 min at a high temperature was found to improve with increasing Si content.

  1. Properties of Native High-Density Lipoproteins Inspire Synthesis of Actively Targeted In Vivo siRNA Delivery Vehicles.

    PubMed

    McMahon, Kaylin M; Plebanek, Michael P; Thaxton, C Shad

    2016-11-15

    Efficient systemic administration of therapeutic short interfering RNA (siRNA) is challenging. High-density lipoproteins (HDL) are natural in vivo RNA delivery vehicles. Specifically, native HDLs: 1) Load single-stranded RNA; 2) Are anionic, which requires charge reconciliation between the RNA and HDL, and 3) Actively target scavenger receptor type B-1 (SR-B1) to deliver RNA. Emphasizing these particular parameters, we employed templated lipoprotein particles (TLP), mimics of spherical HDLs, and self-assembled them with single-stranded complements of, presumably, any highly unmodified siRNA duplex pair after formulation with a cationic lipid. Resulting siRNA templated lipoprotein particles (siRNA-TLP) are anionic and tunable with regard to RNA assembly and function. Data demonstrate that the siRNA-TLPs actively target SR-B1 to potently reduce androgen receptor (AR) and enhancer of zeste homolog 2 (EZH2) proteins in multiple cancer cell lines. Systemic administration of siRNA-TLPs demonstrated no off-target toxicity and significantly reduced the growth of prostate cancer xenografts. Thus, native HDLs inspired the synthesis of a hybrid siRNA delivery vehicle that can modularly load single-stranded RNA complements after charge reconciliation with a cationic lipid, and that function due to active targeting of SR-B1.

  2. Natural Particulates Inspired Specific-Targeted Codelivery of siRNA and Paclitaxel for Collaborative Antitumor Therapy.

    PubMed

    Wang, Ruoning; Zhao, Ziqiang; Han, Yue; Hu, Shihao; Opoku-Damoah, Yaw; Zhou, Jianping; Yin, Lifang; Ding, Yang

    2017-09-05

    The effective combination of drugs promoting antiangiogenesis and apoptosis effects has proven to be a promising collaborative tumor antidote; and the codelivery of small interfering RNA (siRNA) and chemotherapy agents within one efficient vehicle has gained more attention over single regimen administration. Herein, vascular endothelial growth factor specific siRNA (siVEGF) and paclitaxel (PTX) were introduced as therapeutic companions and coencapsulated into naturally mimic high-density lipoproteins (rHDL/siVEGF-PTX), so that various mechanisms of treatment can occur simultaneously. The terminal nanoparticles share capacity of specific-targeting to tumor cells overexpressed scavenger receptor class B type I (SR-BI) and deliver siVEGF and PTX into cytoplasm by a nonendocytosis mechanism. By exchanging HDL core lipids with hydrophobic therapeutics, rHDL/siVEGF-PTX possessed particle size of ∼160 nm, surface potential of ∼-20 mV, and desirable long-term storage stability. In vitro results confirmed that the parallel activity of siVEGF and PTX displayed enhanced anticancer efficacy. The half-maximal inhibitory concentration (IC50) of rHDL/siVEGF-PTX toward human breast cancer MCF-7 cell is 0.26 μg/mL (PTX concentration), which presents a 14.96-fold increase in cytotoxicity by taking Taxol as comparison. Moreover, in vivo results further demonstrated that rHDL/siVEGF-PTX performed enhanced tumor growth inhibition via natural targeting pathway, accompanied by remarkable inhibition of neovascularization in situ caused by siVEGF silencing in down-regulation of VEGF proteins. On the premise of effective drug codelivery, rHDL/siVEGF-PTX demonstrated high tumor targeting for collaborative antitumor efficacy without side effects after systemic administration, and this bioinspired strategy could open an avenue for exploration of combined anticancer therapy.

  3. Co-delivery of doxorubicin and siRNA using octreotide-conjugated gold nanorods for targeted neuroendocrine cancer therapy

    NASA Astrophysics Data System (ADS)

    Xiao, Yuling; Jaskula-Sztul, Renata; Javadi, Alireza; Xu, Wenjin; Eide, Jacob; Dammalapati, Ajitha; Kunnimalaiyaan, Muthusamy; Chen, Herbert; Gong, Shaoqin

    2012-10-01

    A multifunctional gold (Au) nanorod (NR)-based nanocarrier capable of co-delivering small interfering RNA (siRNA) against achaete-scute complex-like 1 (ASCL1) and an anticancer drug (doxorubicin (DOX)) specifically to neuroendocrine (NE) cancer cells was developed and characterized for combined chemotherapy and siRNA-mediated gene silencing. The Au NR was conjugated with (1) DOX, an anticancer drug, via a pH-labile hydrazone linkage to enable pH-controlled drug release, (2) polyarginine, a cationic polymer for complexing siRNA, and (3) octreotide (OCT), a tumor-targeting ligand, to specifically target NE cancer cells with overexpressed somatostatin receptors. The Au NR-based nanocarriers exhibited a uniform size distribution as well as pH-sensitive drug release. The OCT-conjugated Au NR-based nanocarriers (Au-DOX-OCT, targeted) exhibited a much higher cellular uptake in a human carcinoid cell line (BON cells) than non-targeted Au NR-based nanocarriers (Au-DOX) as measured by both flow cytometry and confocal laser scanning microscopy (CLSM). Moreover, Au-DOX-OCT-ASCL1 siRNA (Au-DOX-OCT complexed with ASCL1 siRNA) resulted in significantly higher gene silencing in NE cancer cells than Au-DOX-ASCL1 siRNA (non-targeted Au-DOX complexed with ASCL1 siRNA) as measured by an immunoblot analysis. Additionally, Au-DOX-OCT-ASCL1 siRNA was the most efficient nanocarrier at altering the NE phenotype of NE cancer cells and showed the strongest anti-proliferative effect. Thus, combined chemotherapy and RNA silencing using NE tumor-targeting Au NR-based nanocarriers could potentially enhance the therapeutic outcomes in treating NE cancers.A multifunctional gold (Au) nanorod (NR)-based nanocarrier capable of co-delivering small interfering RNA (siRNA) against achaete-scute complex-like 1 (ASCL1) and an anticancer drug (doxorubicin (DOX)) specifically to neuroendocrine (NE) cancer cells was developed and characterized for combined chemotherapy and siRNA-mediated gene silencing. The

  4. Delivery of kinesin spindle protein targeting siRNA in solid lipid nanoparticles to cellular models of tumor vasculature

    SciTech Connect

    Ying, Bo; Campbell, Robert B.

    2014-04-04

    Highlights: • siRNA-lipid nanoparticles are solid particles not lipid bilayers with aqueous core. • High, but not low, PEG content can prevent nanoparticle encapsulation of siRNA. • PEG reduces cellular toxicity of cationic nanoparticles in vitro. • PEG reduces zeta potential while improving gene silencing of siRNA nanoparticles. • Kinesin spindle protein can be an effective target for tumor vascular targeting. - Abstract: The ideal siRNA delivery system should selectively deliver the construct to the target cell, avoid enzymatic degradation, and evade uptake by phagocytes. In the present study, we evaluated the importance of polyethylene glycol (PEG) on lipid-based carrier systems for encapsulating, and delivering, siRNA to tumor vessels using cellular models. Lipid nanoparticles containing different percentage of PEG were evaluated based on their physical chemical properties, density compared to water, siRNA encapsulation, toxicity, targeting efficiency and gene silencing in vitro. siRNA can be efficiently loaded into lipid nanoparticles (LNPs) when DOTAP is included in the formulation mixture. However, the total amount encapsulated decreased with increase in PEG content. In the presence of siRNA, the final formulations contained a mixed population of particles based on density. The major population which contains the majority of siRNA exhibited a density of 4% glucose, and the minor fraction associated with a decreased amount of siRNA had a density less than PBS. The inclusion of 10 mol% PEG resulted in a greater amount of siRNA associated with the minor fraction. Finally, when kinesin spindle protein (KSP) siRNA was encapsulated in lipid nanoparticles containing a modest amount of PEG, the proliferation of endothelial cells was inhibited due to the efficient knock down of KSP mRNA. The presence of siRNA resulted in the formation of solid lipid nanoparticles when prepared using the thin film and hydration method. LNPs with a relatively modest amount of

  5. Comparing the soft x-rays transport in Si and Ge-sandwich targets by measuring transmission flux

    NASA Astrophysics Data System (ADS)

    Zhang, Lu; Li, Liling; Guo, Liang; Jing, Longfei; Lin, Zhiwei; Qing, Bo; Li, Zhichao; Li, Hang; Kuang, Longyu; Zheng, Jianhua; Zhang, Jiyan; Yang, Jiamin; Jiang, Shaoen; Ding, Yongkun

    2017-03-01

    Mid-Z dopant in ablator is very important in point design targets. In this paper, we develop a method to evaluate the soft x-ray transport of doped material with one dimensional planar target. The targets are designed as sandwich, so that the mid-layer is heated by M band, shock and thermal wave. The transmission fluxes from heated targets are measured with band-pass x-ray diodes. The Si and Ge dopants are evaluated with this method. The experimental results show that 1-1.6 keV x-ray transmission flux through Si-sandwich is higher than that through Ge-sandwich all the time. Also, the comparison of opacities shows that the opacity of Si is lower than that of Ge in the range of 1-1.6 keV, which is consistent with the experimental results.

  6. Dual Tumor-Targeting Nanocarrier System for siRNA Delivery Based on pRNA and Modified Chitosan.

    PubMed

    Li, Lin; Hu, Xiaoqin; Zhang, Min; Ma, Siyu; Yu, Fanglin; Zhao, Shiqing; Liu, Nan; Wang, Zhiyuan; Wang, Yu; Guan, Hua; Pan, Xiujie; Gao, Yue; Zhang, Yue; Liu, Yan; Yang, Yang; Tang, Xuemei; Li, Mingyuan; Liu, Cheng; Li, Zhiping; Mei, Xingguo

    2017-09-15

    Highly specific and efficient delivery of siRNA is still unsatisfactory. Herein, a dual tumor-targeting siRNA delivery system combining pRNA dimers with chitosan nanoparticles (CNPPs) was designed to improve the specificity and efficiency of siRNA delivery. In this dual delivery system, folate-conjugated and PEGylated chitosan nanoparticles encapsulating pRNA dimers were used as the first class of delivery system and would selectively deliver intact pRNA dimers near or into target cells. pRNA dimers simultaneously carrying siRNA and targeting aptamer, the second class of delivery system, would specifically deliver siRNA into the target cells via aptamer-mediated endocytosis or proper particle size. To certify the delivering efficiency of this dual system, CNPPs, pRNA dimers alone, chitosan nanoparticles containing siRNA with folate conjugation and PEGylation (CNPS), and chitosan nanoparticles containing pRNA dimers alone (CN) were first prepared. Then, we observed that treatment with CNPPs resulted in increased cellular uptake, higher cell apoptosis, stronger cell cytotoxicity, and more efficacious gene silencing compared to the other three formulations. Higher accumulation of siRNA in the tumor site, stronger tumor inhibition, and longer circulating time were also observed with CNPPs compared to other formulations. In conclusion, this dual nanocarrier system showed high targeting and favorable therapeutic efficacy both in vitro and in vivo. Thereby, a new approach is provided in this study for specific and efficient delivery of siRNA, which lays a foundation for the development of pRNA hexamers, which can simultaneously carry six different substances. Copyright © 2017 The Author(s). Published by Elsevier Inc. All rights reserved.

  7. Silencing β3 Integrin by Targeted ECO/siRNA Nanoparticles Inhibits EMT and Metastasis of Triple Negative Breast Cancer

    PubMed Central

    Mack, Margaret A.; Schiemann, William P.; Lu, Zheng-Rong

    2015-01-01

    Metastatic breast cancer is the second leading cause of cancer-related deaths amongst women. Triple-negative breast cancer (TNBC) is a highly aggressive subcategory of breast cancer and currently lacks well-defined molecular targets for effective targeted therapies. Disease relapse, metastasis, and drug resistance render standard chemotherapy ineffective in the treatment of TNBC. Since previous studies coupled β3 integrin to epithelial-mesenchymal transition (EMT) and metastasis, we exploited β3 integrin as a therapeutic target to treat TNBC by delivering β3 integrin siRNA via lipid ECO-based nanoparticles (ECO/siβ3). Treatment of TNBC cells with ECO/siβ3 was sufficient to effectively silence β3 integrin expression, attenuate TGF-β-mediated EMT and invasion, restore TGF-β-mediated cytostasis, and inhibit 3-dimensional organoid growth. Modification of ECO/siβ3 nanoparticles with an RGD peptide via a PEG spacer enhanced siRNA uptake by post-EMT cells. Intravenous injections of RGD-targeted ECO/siβ3 nanoparticles in vivo alleviated primary tumor burden, and more importantly, significantly inhibited metastasis. Mice bearing orthotopic, TGF-β-pre-stimulated MDA-MB-231 tumors that were treated with RGD-targeted ECO/siβ3 nanoparticles were free of metastases and relapse after primary tumor resection and 4 weeks after release from the treatment, in comparison to untreated mice. Collectively, these results highlight ECO/siβ3 nanoparticles as a promising therapeutic regimen to combat TNBC. PMID:25858145

  8. Mitochondria apoptosis pathway synergistically activated by hierarchical targeted nanoparticles co-delivering siRNA and lonidamine.

    PubMed

    Zhang, Bing-Feng; Xing, Lei; Cui, Peng-Fei; Wang, Feng-Zhen; Xie, Rong-Lin; Zhang, Jia-Liang; Zhang, Mei; He, Yu-Jing; Lyu, Jin-Yuan; Qiao, Jian-Bin; Chen, Bao-An; Jiang, Hu-Lin

    2015-08-01

    The mitochondria-mediated apoptosis pathway is an effective option for cancer therapy due to the presence of cell-suicide weapons in mitochondria. However, anti-apoptotic proteins that are over-expressed in the mitochondria of many malignant tumors, such as Bcl-2 protein, could allow the cancer cells to evade apoptosis, greatly reducing the efficacy of this type of chemotherapy. Here, we constructed a hierarchical targeted delivery system that can deliver siRNA and chemotherapeutic agents sequentially to tumor cells and mitochondria. In detail, the copolymer TPP-CP-LND (TCPL) was synthesized by the mitochondria-targeting ligand triphenylphosphine (TPP) and therapeutic drug lonidamine (LND) conjugated to the polyethyleneimine in chitosan-graft-PEI (CP), and then complexed with siRNA. Followed, the complexes were coated with poly(acrylic acid)-polyethylene glycol-folic acid (PPF) copolymer to form a hierarchical targeted co-delivery system (TCPL/siRNA/PPF NPs). The TCPL/siRNA/PPF NPs had a neutral surface charge, were stable in plasma and exhibited pH-responsive shell separation. Remarkably, the TCPL/siRNA/PPF NPs simultaneously released siBcl-2 into the cytoplasm and delivered LND to mitochondria in the same cancer cell after FA-directed internalization, and even synergistically activated mitochondria apoptosis pathway. This work demonstrated the potential of RNA-interference and mitochondria-targeted chemotherapeutics to collaboratively stimulate the mitochondria apoptosis pathway for cancer therapy. Copyright © 2015 Elsevier Ltd. All rights reserved.

  9. In Silico Design and Experimental Validation of siRNAs Targeting Conserved Regions of Multiple Hepatitis C Virus Genotypes

    PubMed Central

    ElHefnawi, Mahmoud; Kim, TaeKyu; Kamar, Mona A.; Min, Saehong; Hassan, Nafisa M.; El-Ahwany, Eman; Kim, Heeyoung; Zada, Suher; Amer, Marwa; Windisch, Marc P.

    2016-01-01

    RNA interference (RNAi) is a post-transcriptional gene silencing mechanism that mediates the sequence-specific degradation of targeted RNA and thus provides a tremendous opportunity for development of oligonucleotide-based drugs. Here, we report on the design and validation of small interfering RNAs (siRNAs) targeting highly conserved regions of the hepatitis C virus (HCV) genome. To aim for therapeutic applications by optimizing the RNAi efficacy and reducing potential side effects, we considered different factors such as target RNA variations, thermodynamics and accessibility of the siRNA and target RNA, and off-target effects. This aim was achieved using an in silico design and selection protocol complemented by an automated MysiRNA-Designer pipeline. The protocol included the design and filtration of siRNAs targeting highly conserved and accessible regions within the HCV internal ribosome entry site, and adjacent core sequences of the viral genome with high-ranking efficacy scores. Off-target analysis excluded siRNAs with potential binding to human mRNAs. Under this strict selection process, two siRNAs (HCV353 and HCV258) were selected based on their predicted high specificity and potency. These siRNAs were tested for antiviral efficacy in HCV genotype 1 and 2 replicon cell lines. Both in silico-designed siRNAs efficiently inhibited HCV RNA replication, even at low concentrations and for short exposure times (24h); they also exceeded the antiviral potencies of reference siRNAs targeting HCV. Furthermore, HCV353 and HCV258 siRNAs also inhibited replication of patient-derived HCV genotype 4 isolates in infected Huh-7 cells. Prolonged treatment of HCV replicon cells with HCV353 did not result in the appearance of escape mutant viruses. Taken together, these results reveal the accuracy and strength of our integrated siRNA design and selection protocols. These protocols could be used to design highly potent and specific RNAi-based therapeutic oligonucleotide

  10. Efficient VEGF targeting delivery of DOX using Bevacizumab conjugated SiO2@LDH for anti-neuroblastoma therapy.

    PubMed

    Zhu, Rongrong; Wang, Zhaoqi; Liang, Peng; He, Xiaolie; Zhuang, Xizhen; Huang, Ruiqi; Wang, Mei; Wang, Qigang; Qian, Yechang; Wang, Shilong

    2017-09-18

    Vascular endothelial growth factor (VEGF) plays an important role in angiogenesis and is highly expressed in carcinoma, which make it an important target for tumor targeting therapy. Neuroblastoma is the main cause for cancer-related death in children. Like most solid tumors, it is also accompanied with the overexpression of VEGF. Doxorubicin Hydrochloride (DOX), a typical chemotherapeutic agent, exhibits efficient anticancer activities for various cancers. However, DOX, without targeting ability, usually causes severe damage to normal tissues. To overcome the shortages, we designed a novel nano-composite, which is Bevacizumab (Bev) modified SiO2@LDH nanoparticles (SiO2@LDH-Bev), loading with DOX to achieve targeting ability and curative efficiency. SiO2@LDH-DOX and SiO2@LDH-Bev-DOX nanoparticles were synthesized and the physicochemical properties were characterized by TEM detection, Zeta potential analysis, FTIR, Raman and XPS analysis. Then in vitro and in vivo anti-neuroblastoma efficiency, targeting ability and mechanisms of anti-carcinoma and anti-angiogenesis of SiO2@LDH-Bev-DOX were explored. Our results indicated that we obtained the core-shell structure SiO2@LDH-Bev with an average diameter of 253±10nm and the amount of conjugated Bev was 4.59±0.38μg/mg SiO2@LDH-Bev. SiO2@LDH-Bev-DOX could improve the cellular uptake and the targeting effect of DOX to brain and tumor, enhance the anti-neuroblastoma and anti-angiogenesis efficiency both in vitro and in vivo, and alleviate side effects of DOX sharply, especially hepatic injury. In addition, we also demonstrated that angiogenesis inhibitory effect was mediated by DOX and VEGF triggered signal pathways, including PI3K/Akt, Raf/MEK/ERK, and adhesion related pathways. In summary, SiO2@LDH-Bev could be a potential VEGF targeting nanocarrier applied in VEGF positive cancer therapy. This paper explored that a novel core-shell structure nanomaterial SiO2@LDH and modified SiO2@LDH with Bevacizumab (Bev) to form a

  11. Delivery of siRNA using ternary complexes containing branched cationic peptides: the role of peptide sequence, branching and targeting.

    PubMed

    Kudsiova, Laila; Welser, Katharina; Campbell, Frederick; Mohammadi, Atefeh; Dawson, Natalie; Cui, Lili; Hailes, Helen C; Lawrence, M Jayne; Tabor, Alethea B

    2016-03-01

    Ternary nanocomplexes, composed of bifunctional cationic peptides, lipids and siRNA, as delivery vehicles for siRNA have been investigated. The study is the first to determine the optimal sequence and architecture of the bifunctional cationic peptide used for siRNA packaging and delivery using lipopolyplexes. Specifically three series of cationic peptides of differing sequence, degrees of branching and cell-targeting sequences were co-formulated with siRNA and vesicles prepared from a 1 : 1 molar ratio of the cationic lipid DOTMA and the helper lipid, DOPE. The level of siRNA knockdown achieved in the human alveolar cell line, A549-luc cells, in both reduced serum and in serum supplemented media was evaluated, and the results correlated to the nanocomplex structure (established using a range of physico-chemical tools, namely small angle neutron scattering, transmission electron microscopy, dynamic light scattering and zeta potential measurement); the conformational properties of each component (circular dichroism); the degree of protection of the siRNA in the lipopolyplex (using gel shift assays) and to the cellular uptake, localisation and toxicity of the nanocomplexes (confocal microscopy). Although the size, charge, structure and stability of the various lipopolyplexes were broadly similar, it was clear that lipopolyplexes formulated from branched peptides containing His-Lys sequences perform best as siRNA delivery agents in serum, with protection of the siRNA in serum balanced against efficient release of the siRNA into the cytoplasm of the cell.

  12. Improvement of cytomegalovirus pp65 DNA vaccine efficacy by co-administration of siRNAs targeting BAK and BAX.

    PubMed

    Liu, Jixiao; Feng, Keke; Zhao, Lu; Luo, Haining; Zhu, Yingjun

    2017-06-01

    The efficacy of DNA vaccines may be improved by small interfering (si)RNA adjuvants targeting pro-apoptotic genes. The aim of the present study was to investigate the capacity of siRNAs targeting B-cell lymphoma 2 homologous antagonist killer (BAK) and B-cell lymphoma 2-associated X protein (BAX) to improve the efficacy of a cytomegalovirus (CMV) vaccine. BALB/c mice were divided into four groups (n=18 in each): unimmunized and immunized with pcDNA 3.1-pp65 expressing CMV 65 kDa matrix phosphoprotein and BAK + BAX siRNAs, pcDNA 3.1-pp65 and control siRNA, or control pcDNA 3.1 and BAK + BAX siRNAs. Immunizations were performed twice with an interval of 3 weeks. CMV-specific mouse splenocyte interferon (IFN)-γ secretion was assessed by ELISPOT; furthermore, an in vivo cytotoxic T lymphocyte assay was performed 2 weeks after the last immunization. After lethal CMV challenge of the mice, body weight, virus titers in the spleens and salivary glands as well as survival were recorded. The amount of splenocytes secreting IFN-γ in response to CMV pp65 peptides and specific lysis of peptide-pulsed target cells were significantly higher in mice administered pcDNA3.1-pp65 and BAK + BAX siRNAs than those in mice administered pcDNA3.1-pp65 and control siRNA (P<0.05 for each). After the virus challenge, the virus titers in the spleens and salivary glands of mice given pcDNA3.1-pp65 and BAK + BAX siRNAs were significantly lower than those in mice immunized with pcDNA3.1-pp65 and control siRNA (P<0.05 for each). Furthermore, mice immunized with pcDNA 3.1-pp65 and control siRNA or BAK + BAX siRNAs survived for longer, and at 21 days after lethal CMV challenge, 66 and 100% of these mice survived, respectively. These mice also experienced less weight loss compared with mice immunized with pcDNA3.1-pp65 and control siRNA (P<0.05). In conclusion, intradermal administration of siRNAs targeting BAK and BAX improved the efficacy of CMV pp65 DNA vaccine.

  13. Dual peptide-mediated targeted delivery of bioactive siRNAs to oral cancer cells in vivo.

    PubMed

    Alexander-Bryant, Angela A; Zhang, Haiwen; Attaway, Christopher C; Pugh, William; Eggart, Laurence; Sansevere, Robert M; Andino, Lourdes M; Dinh, Lu; Cantini, Liliana P; Jakymiw, Andrew

    2017-09-01

    Despite significant advances in cancer treatment, the prognosis for oral cancer remains poor in comparison to other cancer types, including breast, skin, and prostate. As a result, more effective therapeutic modalities are needed for the treatment of oral cancer. Consequently, in the present study, we examined the feasibility of using a dual peptide carrier approach, combining an epidermal growth factor receptor (EGFR)-targeting peptide with an endosome-disruptive peptide, to mediate targeted delivery of small interfering RNAs (siRNAs) into EGFR-overexpressing oral cancer cells and induce silencing of the targeted oncogene, cancerous inhibitor of protein phosphatase 2A (CIP2A). Fluorescence microscopy, real-time PCR, Western blot analysis, and in vivo bioimaging of mice containing orthotopic xenograft tumors were used to examine the ability of the dual peptide carrier to mediate specific delivery of bioactive siRNAs into EGFR-overexpressing oral cancer cells/tissues. Co-complexation of the EGFR-targeting peptide, GE11R9, with the endosome-disruptive 599 peptide facilitated the specific uptake of siRNAs into oral cancer cells overexpressing EGFR in vitro with optimal gene silencing observed at a 60:30:1 (GE11R9:599:siRNA) molar ratio. Furthermore, when administered systemically to mice bearing xenograft oral tumors, this dual peptide complex mediated increased targeted delivery of siRNAs into tumor tissues in comparison to the 599 peptide alone and significantly enhanced CIP2A silencing. Herein we provide the first report demonstrating the clinical potential of a dual peptide strategy for siRNA-based therapeutics by synergistically mediating the effective targeting and delivery of bioactive siRNAs into EGFR-overexpressing oral cancer cells. Copyright © 2017 Elsevier Ltd. All rights reserved.

  14. Monocrystalline silicon gradiometer for gravity experiments in space

    NASA Technical Reports Server (NTRS)

    Richard, Jean-Paul

    1987-01-01

    A very important research effort has been made in the last decade in the field of high precision measurement with laser instrumentation. The development of a space borne gradiometer operating at a high sensitivity level using laser measurement of the distance between proof mass over a short base line of order one meter is discussed. Two aspects of laser technology make it a promising tool for gradiometry measurements: quantum limited accuracy and absolute distance measurements. The quantum limit associated with laser instrumentation was formulated. The relevant quantum and classical sources of errors in laser measurements were reviewed and corresponding laser performance requirements for gradient measurements were evaluated. Some mechanical quality factor measurements were made on simple resonant monocrystalline silicon suspensions. It was discovered that the use of zero derivative restoring forces to position the gradiometer test masses will dramatically reduce the gradiometer temperature control requirements. A laser beam side injection scheme was discovered which permits rejection of common mode accelerations. These concepts are briefly discussed.

  15. Large-area monocrystalline silicon thin films by annealing of macroporous arrays: Understanding and tackling defects in the material

    NASA Astrophysics Data System (ADS)

    Depauw, Valérie; Gordon, Ivan; Beaucarne, Guy; Poortmans, Jef; Mertens, Robert; Celis, Jean-Pierre

    2009-08-01

    A concept that could provide a thin monocrystalline-silicon absorber layer without resorting to the expensive step of epitaxy would be very appealing for reducing the cost of solar cells. The empty-space-in-silicon technique by which thin films of silicon can be formed by reorganization of regular arrays of cylindrical voids at high temperature may be such a concept if the high quality of the thin film could be ensured on centimeter-large areas. While previous works mainly investigated the influence of the porous array on the final structure, this work focuses on the practical aspects of the high-temperature step and its application to large areas. An insight into the defects that may form is given and the origin of these defects is discussed, providing recommendations on how to avoid them. Surface roughening, pitting, formation of holes, and silicon pillars could be attributed to the nonuniform reactions between Si, SiO2, and SiO. Hydrogen atmospheres are therefore preferred for reorganization of macroporous arrays. Argon atmospheres, however, may provide high-quality silicon thin films as well, possibly even more easily transferable, as long as annealing is performed in controlled, clean, and oxygen-free conditions. Our experiments on large areas also highlight the importance of kinetics, which had not been considered up to now and which will require further understanding to ensure a complete reorganization over any wafer area.

  16. Immunostimulatory motifs enhance antiviral siRNAs targeting highly pathogenic avian influenza H5N1.

    PubMed

    Stewart, Cameron R; Karpala, Adam J; Lowther, Sue; Lowenthal, John W; Bean, Andrew G

    2011-01-01

    Highly pathogenic avian influenza (HPAI) H5N1 virus is endemic in many regions around the world and remains a significant pandemic threat. To date H5N1 has claimed almost 300 human lives worldwide, with a mortality rate of 60% and has caused the death or culling of hundreds of millions of poultry since its initial outbreak in 1997. We have designed multi-functional RNA interference (RNAi)-based therapeutics targeting H5N1 that degrade viral mRNA via the RNAi pathway while at the same time augmenting the host antiviral response by inducing host type I interferon (IFN) production. Moreover, we have identified two factors critical for maximising the immunostimulatory properties of short interfering (si)RNAs in chicken cells (i) mode of synthesis and (ii) nucleoside sequence to augment the response to virus. The 5-bp nucleoside sequence 5'-UGUGU-3' is a key determinant in inducing high levels of expression of IFN-α, -β, -λ and interleukin 1-β in chicken cells. Positioning of this 5'-UGUGU-3' motif at the 5'-end of the sense strand of siRNAs, but not the 3'-end, resulted in a rapid and enhanced induction of type I IFN. An anti-H5N1 avian influenza siRNA directed against the PB1 gene (PB1-2257) tagged with 5'-UGUGU-3' induced type I IFN earlier and to a greater extent compared to a non-tagged PB1-2257. Tested against H5N1 in vitro, the tagged PB1-2257 was more effective than non-tagged PB1-2257. These data demonstrate the ability of an immunostimulatory motif to improve the performance of an RNAi-based antiviral, a finding that may influence the design of future RNAi-based anti-influenza therapeutics.

  17. Tumor targeting RGD conjugated bio-reducible polymer for VEGF siRNA expressing plasmid delivery.

    PubMed

    Kim, Hyun Ah; Nam, Kihoon; Kim, Sung Wan

    2014-08-01

    Targeted delivery of therapeutic genes to the tumor site is critical for successful and safe cancer gene therapy. The arginine grafted bio-reducible poly (cystamine bisacrylamide-diaminohexane, CBA-DAH) polymer (ABP) conjugated poly (amido amine) (PAMAM), PAM-ABP (PA) was designed previously as an efficient gene delivery carrier. To achieve high efficacy in cancer selective delivery, we developed the tumor targeting bio-reducible polymer, PA-PEG1k-RGD, by conjugating cyclic RGDfC (RGD) peptides, which bind αvβ3/5 integrins, to the PAM-ABP using polyethylene glycol (PEG, 1 kDa) as a spacer. Physical characterization showed nanocomplex formation with bio-reducible properties between PA-PEG1k-RGD and plasmid DNA (pDNA). In transfection assays, PA-PEG1k-RGD showed significantly higher transfection efficiency in comparison with PAM-ABP or PA-PEG1k-RAD in αvβ3/5 positive MCF7 breast cancer and PANC-1 pancreatic cancer cells. The targeting ability of PA-PEG1k-RGD was further established using a competition assay. To confirm the therapeutic effect, the VEGF siRNA expressing plasmid was constructed and then delivered into cancer cells using PA-PEG1k-RGD. PA-PEG1k-RGD showed 20-59% higher cellular uptake rate into MCF7 and PANC-1 than that of non-targeted polymers. In addition, MCF7 and PANC-1 cancer cells transfected with PA-PEG1k-RGD/pshVEGF complexes had significantly decreased VEGF gene expression (51-71%) and cancer cell viability (35-43%) compared with control. These results demonstrate that a tumor targeting bio-reducible polymer with an anti-angiogenic therapeutic gene could be used for efficient and safe cancer gene therapy. Copyright © 2014 Elsevier Ltd. All rights reserved.

  18. Tumor targeting RGD conjugated bio-reducible polymer for VEGF siRNA expressing plasmid delivery

    PubMed Central

    Kim, Hyun Ah; Nam, Kihoon; Kim, Sung Wan

    2014-01-01

    Targeted delivery of therapeutic genes to the tumor site is critical for successful and safe cancer gene therapy. The arginine grafted bio-reducible poly (cystamine bisacrylamide-diaminohexane, CBA-DAH) polymer (ABP) conjugated poly (amido amine) (PAMAM), PAM-ABP (PA) was designed previously as an efficient gene delivery carrier. To achieve high efficacy in cancer selective delivery, we developed the tumor targeting bio-reducible polymer, PA-PEG1k-RGD, by conjugating cyclic RGDfC (RGD) peptides, which bind αvβ3/5 integrins, to the PAM-ABP using polyethylene glycol (PEG,1kDa) as a spacer. Physical characterization showed nanocomplex formation with bio-reducible properties between PA-PEG1k-RGD and plasmid DNA (pDNA). In transfection assays, PA-PEG1k-RGD showed significantly higher transfection efficiency in comparison with PAM-ABP or PA-PEG1k-RGD in αvβ3/5 positive MCF7 breast cancer and PANC-1 pancreatic cancer cells. The targeting ability of PA-PEG1k-RGD was further established using a competition assay. To confirm the therapeutic effect, the VEGF siRNA expressing plasmid was constructed and then delivered into cancer cells using PA-PEG1k-RGD. PA-PEG1k-RGD showed 20-59% higher cellular uptake rate into MCF7 and PANC-1 than that of non-targeted polymers. In addition, MCF7 and PANC-1 cancer cells transfected with PA-PEG1k-RGD/pshVEGF complexes had significantly decreased VEGF gene expression (51-71%) and cancer cell viability (35-43%) compared with control. These results demonstrate that a tumor targeting bio-reducible polymer with an anti-angiogenic therapeutic gene could be used for efficient and safe cancer gene therapy. PMID:24894645

  19. PEG grafting of polyethylenimine (PEI) exerts different effects on DNA transfection and siRNA-induced gene targeting efficacy.

    PubMed

    Malek, Anastasia; Czubayko, Frank; Aigner, Achim

    2008-02-01

    Gene targeting by RNA interference (RNAi) is mediated through small interfering RNA (siRNA), which, as plasmid DNA molecules, can be delivered into cells by polyethylenimines (PEI). Grafting with poly(ethylene glycol) has been introduced previously to improve PEI biocompatibility; however, data on the effects of PEGylation have been somewhat contradictory and various PEI(-PEG) need to be evaluated independently for DNA transfection and siRNA gene targeting efficacies. We directly compare plasmid DNA transfection and siRNA-mediated gene targeting efficacies, employing a larger set of polyethylenimine-graft-poly(ethylene glycol) (PEI-g-PEG; PEI(-PEG)) with different molecular weights and degrees of PEG substitution. We performed tissue culture-based bioassays on DNA transfection and siRNA-mediated targeting efficacies as well as on toxicity and cellular nucleic acid uptake, and, using sensitive assays based on radioactive labelling, physicochemically characterize the complexes regarding the degree of nucleic acid complexation and complex stabilities under various conditions. In contrast to the DNA transfection efficacy, siRNA-mediated gene targeting is much less dependent on the PEGylation of PEI or on the N/P ( = PEI nitrogen/nucleic acid phosphate) ratio. A more detailed analysis reveals that, in order to define optimal N/P ratios for DNA transfection, complex toxicities and nucleic acid uptake are the most critical parameters. In contrast, at optimal N/P ratios, complex stabilities and complexation efficacies determine PEI(-PEG)/DNA transfection efficacies and the major differences between various PEI(-PEG) are observed. All these parameters are less critical for PEI(-PEG)/siRNA gene targeting efficacy. Thus, our data lead to the distinction between three PEI(-PEG) groups, which relies on the differences in transfection rather than gene targeting efficacies, and which is correlated with the molecular weights and degrees of PEG substitution. In contrast to PEI

  20. Orally delivered thioketal nanoparticles loaded with TNF-α-siRNA target inflammation and inhibit gene expression in the intestines

    NASA Astrophysics Data System (ADS)

    Wilson, D. Scott; Dalmasso, Guillaume; Wang, Lixin; Sitaraman, Shanthi V.; Merlin, Didier; Murthy, Niren

    2010-11-01

    Small interfering RNAs (siRNAs) directed against proinflammatory cytokines have the potential to treat numerous diseases associated with intestinal inflammation; however, the side-effects caused by the systemic depletion of cytokines demands that the delivery of cytokine-targeted siRNAs be localized to diseased intestinal tissues. Although various delivery vehicles have been developed to orally deliver therapeutics to intestinal tissue, none of these strategies has demonstrated the ability to protect siRNA from the harsh environment of the gastrointestinal tract and target its delivery to inflamed intestinal tissue. Here, we present a delivery vehicle for siRNA, termed thioketal nanoparticles (TKNs), that can localize orally delivered siRNA to sites of intestinal inflammation, and thus inhibit gene expression in inflamed intestinal tissue. TKNs are formulated from a polymer, poly-(1,4-phenyleneacetone dimethylene thioketal), that degrades selectively in response to reactive oxygen species (ROS). Therefore, when delivered orally, TKNs release siRNA in response to the abnormally high levels of ROS specific to sites of intestinal inflammation. Using a murine model of ulcerative colitis, we demonstrate that orally administered TKNs loaded with siRNA against the proinflammatory cytokine tumour necrosis factor-alpha (TNF-α) diminish TNF-α messenger RNA levels in the colon and protect mice from ulcerative colitis.

  1. Noninvasive Drug Delivery Using Ultrasound: Targeting Melanoma Using siRNA Against Mutant (V600E) B-Raf

    NASA Astrophysics Data System (ADS)

    Tran, Melissa A.; Gowda, Raghavendra; Park, Eun-Joo; Adair, James; Smith, Nadine; Kester, Mark; Robertson, Gavin P.

    2009-04-01

    Melanoma is the most deadly form of skin cancer. Currently early surgical removal is the best treatment option for melanoma patients with little hope of successful treatment of late stage melanoma. Clearly new treatment options must be explored. Topical administration of drugs provides the advantage of being able to apply large quantities of drug in close proximity to the tumor without the issue of systemic side effects. However, the natural barrier formed by the skin must first be overcome for topical treatment to become a viable option. With this in mind we have sought to use low-frequency ultrasound to transiently permeabilize the stratum corneum and successfully deliver liposomal siRNA to melanoma cells residing at the basement membrane. B-Raf is one of the most frequently activated genes in melanoma, making it an ideal candidate for targeting via siRNA. The novel liposomes used in this study load siRNA, protect if from the outside environment and lead to knockdown of target message. Combining ultrasound with liposomal siRNA we show that siRNA can be delivered into melanoma cells. Additionally, we show that siRNA to mutant B-Raf can effectively inhibit melanoma growth in reconstructs and in mice by 60% and 30% respectively. Therefore, ultrasound with liposomal siRNA is a potentially valuable treatment option for melanoma patients.

  2. siRNA screen identifies QPCT as a druggable target for Huntington's disease.

    PubMed

    Jimenez-Sanchez, Maria; Lam, Wun; Hannus, Michael; Sönnichsen, Birte; Imarisio, Sara; Fleming, Angeleen; Tarditi, Alessia; Menzies, Fiona; Ed Dami, Teresa; Xu, Catherine; Gonzalez-Couto, Eduardo; Lazzeroni, Giulia; Heitz, Freddy; Diamanti, Daniela; Massai, Luisa; Satagopam, Venkata P; Marconi, Guido; Caramelli, Chiara; Nencini, Arianna; Andreini, Matteo; Sardone, Gian Luca; Caradonna, Nicola P; Porcari, Valentina; Scali, Carla; Schneider, Reinhard; Pollio, Giuseppe; O'Kane, Cahir J; Caricasole, Andrea; Rubinsztein, David C

    2015-05-01

    Huntington's disease (HD) is a currently incurable neurodegenerative condition caused by an abnormally expanded polyglutamine tract in huntingtin (HTT). We identified new modifiers of mutant HTT toxicity by performing a large-scale 'druggable genome' siRNA screen in human cultured cells, followed by hit validation in Drosophila. We focused on glutaminyl cyclase (QPCT), which had one of the strongest effects on mutant HTT-induced toxicity and aggregation in the cell-based siRNA screen and also rescued these phenotypes in Drosophila. We found that QPCT inhibition induced the levels of the molecular chaperone αB-crystallin and reduced the aggregation of diverse proteins. We generated new QPCT inhibitors using in silico methods followed by in vitro screening, which rescued the HD-related phenotypes in cell, Drosophila and zebrafish HD models. Our data reveal a new HD druggable target affecting mutant HTT aggregation and provide proof of principle for a discovery pipeline from druggable genome screen to drug development.

  3. Non-Condensing Polymeric Nanoparticles for Targeted Gene and siRNA Delivery

    PubMed Central

    Xu, Jing; Ganesh, Shanthi; Amiji, Mansoor

    2011-01-01

    Gene therapy has shown a tremendous potential to benefit patients in a variety of disease conditions. However, finding a safe and effective systemic delivery system is the major obstacle in this area. Although viral vectors showed promise for high transfection rate, the immunogenicity associated with these systems has hindered further development. As an alternative to viral gene delivery, this review focuses on application of novel safe and effective non-condensing polymeric systems that have shown high transgene expression when administered systemically or by the oral route. Type B gelatin-based engineered nanocarriers were evaluated for passive and active tumor-targeted delivery and transfection using both reporter and therapeutic plasmid DNA. Additionally, we have shown that nanoparticles-in-microsphere oral system (NiMOS) can efficiently deliver reporter and therapeutic gene constructs in the gastrointestinal tract. Additionally, there has been a significant recent interest in the use small interfering RNA (siRNA) as a therapeutic system for gene silencing. Both gelatin nanoparticles and NiMOS have shown activity in systemic and oral delivery of siRNA, respectively. PMID:21621597

  4. siRNA screen identifies QPCT as a druggable target for Huntington’s disease

    PubMed Central

    Jimenez-Sanchez, Maria; Lam, Wun; Tarditi, Alessia; Menzies, Fiona; Dami, Teresa Ed; Xu, Catherine; Gonzalez-Couto, Eduardo; Lazzeroni, Giulia; Heitz, Freddy; Diamanti, Daniela; Massai, Luisa; Satagopam, Venkata P.; Marconi, Guido; Caramelli, Chiara; Nencini, Arianna; Andreini, Matteo; Sardone, Gian Luca; Caradonna, Nicola P.; Porcari, Valentina; Scali, Carla; Schneider, Reinhard; Pollio, Giuseppe; O’Kane, Cahir J.; Caricasole, Andrea; Rubinsztein, David C.

    2015-01-01

    Huntington’s disease (HD) is a currently incurable neurodegenerative condition caused by an abnormally expanded polyglutamine tract in huntingtin (HTT). We identified novel modifiers of mutant HTT toxicity by performing a large-scale “druggable genome” siRNA screen in human cultured cells, followed by hit validation in Drosophila. We focused on glutaminyl cyclase (QPCT), which had one of the strongest effects on mutant HTT-induced toxicity and aggregation in the cell-based siRNA screen, and which also rescued these phenotypes in Drosophila. We found that QPCT inhibition induced the levels of the molecular chaperone alpha B-crystallin and reduced the aggregation of diverse proteins. We generated novel QPCT inhibitors using in silico methods followed by in vitro screens, which rescued the HD-related phenotypes in cell, Drosophila and zebrafish HD models. Our data reveal a novel HD druggable target affecting mutant huntingtin aggregation, and provide proof-of-principle for a discovery pipeline from druggable genome screen to drug development. PMID:25848931

  5. Anti-HBV efficacy of combined siRNAs targeting viral gene and heat shock cognate 70

    PubMed Central

    2012-01-01

    Background Hepatitis B virus (HBV) infection is a major health concern with more than two billion individuals currently infected worldwide. Because of the limited effectiveness of existing vaccines and drugs, development of novel antiviral strategies is urgently needed. Heat stress cognate 70 (Hsc70) is an ATP-binding protein of the heat stress protein 70 family. Hsc70 has been found to be required for HBV DNA replication. Here we report, for the first time, that combined siRNAs targeting viral gene and siHsc70 are highly effective in suppressing ongoing HBV expression and replication. Methods We constructed two plasmids (S1 and S2) expressing short hairpin RNAs (shRNAs) targeting surface open reading frame of HBV(HBVS) and one plasmid expressing shRNA targeting Hsc70 (siHsc70), and we used the EGFP-specific siRNA plasmid (siEGFP) as we had previously described. First, we evaluated the gene-silencing efficacy of both shRNAs using an enhanced green fluorescent protein (EGFP) reporter system and flow cytometry in HEK293 and T98G cells. Then, the antiviral potencies of HBV-specific siRNA (siHBV) in combination with siHsc70 in HepG2.2.15 cells were investigated. Moreover, type I IFN and TNF-α induction were measured by quantitative real-time PCR and ELISA. Results Cotransfection of either S1 or S2 with an EGFP plasmid produced an 80%–90% reduction in EGFP signal relative to the control. This combinational RNAi effectively and specifically inhibited HBV protein, mRNA and HBV DNA, resulting in up to a 3.36 log10 reduction in HBV load in the HepG2.2.15 cell culture supernatants. The combined siRNAs were more potent than siHBV or siHsc70 used separately, and this approach can enhance potency in suppressing ongoing viral gene expression and replication in HepG2.2.15 cells while forestalling escape by mutant HBV. The antiviral synergy of siHBV used in combination with siHsc70 produced no cytotoxicity and induced no production of IFN-α, IFN-β and TNF-α in transfected

  6. Influence of SiRNA targeting survivin on chemosensitivity of H460/cDDP lung cancer cells.

    PubMed

    Yang, H; Fu, J-H; Hu, Y; Huang, W-Z; Zheng, B; Wang, G; Zhang, X; Wen, J

    2008-01-01

    We investigated the difference in survivin expression between a multidrug-resistant lung cancer cell line (H460/cDDP) and its parental counterpart (H460) and the influence of siRNA targeting survivin on the chemosensitivity of H460/cDDP. SiRNA targeting survivin was transfected into H460/cDDP cells using a liposome approach. Survivin mRNA and protein expression were significantly higher in H460/cDDP than H460 cells. The median inhibitory concentrations (IC(50)s) for cisplatin and paclitaxol in vitro against H460/cDDP cells were significantly lower in cells treated with survivin-specific siRNA than in control cells. Apoptosis and cleaved caspase-3 expression were analysed using annexin V and Western blotting, respectively, and showed a significant increase in apoptosis after treatment with the chemotherapeutic agents plus specific siRNA. Specific siRNA sensitized H460/cDDP cells to both cisplatin and paclitaxol. Thus, survivin appears to participate in the multidrug resistance mechanism of H460/cDDP cells and siRNA targeting survivin has the potential to increase the sensitivity of drug-resistant cancer cells to anticancer drugs.

  7. Nanoparticle-formulated siRNA targeting integrins inhibits hepatocellular carcinoma progression in mice

    PubMed Central

    Bogorad, Roman L; Yin, Hao; Zeigerer, Anja; Nonaka, Hidenori; Ruda, Vera; Zerial, Marino; Anderson, Daniel G; Koteliansky, Victor

    2014-01-01

    Integrins play an important role during development, regulating cell differentiation, proliferation and survival. Here we show that knockdown of integrin subunits slows down the progression of hepatocellular carcinoma (HCC). Using nanoparticulate delivery of short interfering RNAs targeting β1 and αv integrin subunits we downregulate all integrin receptors in hepatocytes. Short-term integrin knockdown (two weeks) does not cause apparent structural or functional perturbations of normal liver tissue. Alterations in liver morphology accumulate upon sustained integrin downregulation (seven weeks). The integrin knockdown leads to significant retardation of HCC progression, reducing proliferation and increasing tumour cell death. This tumour retardation is accompanied by reduced activation of MET oncogene as well as expression of its mature form on the cell surface. Our data suggest that transformed proliferating cells from HCC are more sensitive to knockdown of integrins than normal quiescent hepatocytes, highlighting the potential of siRNA-mediated inhibition of integrins as an anti-cancer therapeutic approach. PMID:24844798

  8. Iron-Oxide-Based Nanovector for Tumor Targeted siRNA Delivery in an Orthotopic Hepatocellular Carcinoma Xenograft Mouse Model.

    PubMed

    Wang, Kui; Kievit, Forrest M; Sham, Jonathan G; Jeon, Mike; Stephen, Zachary R; Bakthavatsalam, Arvind; Park, James O; Zhang, Miqin

    2016-01-27

    Hepatocellular carcinoma (HCC) is one of the deadliest cancers worldwide. Small interfering RNA (siRNA) holds promise as a new class of therapeutics for HCC, as it can achieve sequence-specific gene knockdown with low cytotoxicity. However, the main challenge in the clinical application of siRNA lies in the lack of effective delivery approaches that need to be highly specific and thus incur low or no systemic toxicity. Here, a nonviral nanoparticle-based gene carrier is presented that can specifically deliver siRNA to HCC. The nanovector (NP-siRNA-GPC3 Ab) is made of an iron oxide core coated with chitosan-polyethylene glycol (PEG) grafted polyethyleneimine copolymer, which is further functionalized with siRNA and conjugated with a monoclonal antibody (Ab) against human glypican-3 (GPC3) receptor highly expressed in HCC. A rat RH7777 HCC cell line that coexpresses human GPC3 and firefly luciferase (Luc) is established to evaluate the nanovector. The nanoparticle-mediated delivery of siRNA against Luc effectively suppresses Luc expression in vitro without notable cytotoxicity. Significantly, NP-siLuc-GPC3 Ab administered intravenously in an orthotopic model of HCC is able to specifically bound to tumor and induce remarkable inhibition of Luc expression. The findings demonstrate the potential of using this nanovector for targeted delivery of therapeutic siRNA to HCC. © 2015 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  9. Targeting (V600E) B-Raf and Akt3 using Nanoliposomal-siRNA Inhibits Cutaneous Melanocytic Lesion Development

    PubMed Central

    Tran, Melissa A.; Gowda, Raghavendra; Sharma, Arati; Park, E. J.; Adair, James; Kester, Mark; Smith, Nadine Barrie; Robertson, Gavin P.

    2013-01-01

    Most events promoting early melanoma development are yet to be identified but deregulation of the B-Raf and Akt3 signaling cascades are important regulators of this process. Approximately 90% of normal moles and ~60% of early invasive cutaneous melanomas contain a T1799A B-Raf mutation (V600EB-Raf), leading to 10X higher enzyme activity and constitutive activation of the MAP kinase pathway. Furthermore, ~70% of melanomas have elevated Akt3 signaling due to increased gene copy number and PTEN loss. Therefore, targeting V600EB-Raf and Akt3 signaling is necessary to prevent or treat cutaneous melanocytic lesions. Agents specifically targeting these proteins are needed, having fewer side effects than those inhibiting both normal and mutant B-Raf protein or targeting all three Akt isoforms. In this study, a unique nanoliposomal-ultrasound mediated approach has been developed for delivering siRNA specifically targeting V600EB-Raf and Akt3 into melanocytic tumors present in skin to retard melanoma development. Novel cationic nanoliposomes stably encapsulate siRNA targeting V600EB-Raf or Akt3, providing protection from degradation and facilitating entry into melanoma cells to decrease expression of these proteins. Low-frequency ultrasound using a lightweight 4-cymbal transducer array enables penetration of nanoliposomal-siRNA complex throughout epidermal and dermal layers of laboratory-generated or animal skin. Nanoliposomal-mediated siRNA targeting of V600EB-Raf and Akt3 led to a cooperatively acting ~65% decrease in early or invasive cutaneous melanoma compared to inhibition of each singly with negligible associated systemic toxicity. Thus, cationic nanoliposomes loaded with siRNA targeting V600EB-Raf and Akt3 provide an effective approach for targeted inhibition of early or invasive cutaneous melanomas. PMID:18794153

  10. Plastic Deformation, Amorphization, and Phase Changes in Extreme Laser Shock Compression of Ta and Si

    NASA Astrophysics Data System (ADS)

    Meyers, Marc

    2015-06-01

    High-amplitude pulsed lasers probe the response of materials at pressures up to 100s of GPa and strain rates of 108 s-1, revealing plastic deformation, phase transformations, and amorphization. Molecular dynamics simulations provide modeling at comparable strain rates and time durations. Shock compression of monocrystalline Ta reveals dislocations at low pressures and twinning at higher pressures (above 24 GPa). Results are compared with predictions from homogeneous dislocation generation and multiplication and the latter mechanism is dominant. The formation of an Omega phase was observed in monocrystalline tantalum at a shock amplitude of approximately 70 GPa. The shear stresses may play a role in the transformation. As shock energy increases, the following structural changes in monocrystalline Si are observed: dislocations and stacking faults; bands of amorphized Si forming on crystallographic orientations consistent with slip; broad regions of amorphized Si; nanocrystalline Si resulting from re-crystallization. MD simulations display similar amorphous regions. Funding: UCOP and DOE SSAP.

  11. High-density fluids and the growth of monocrystalline diamonds

    NASA Astrophysics Data System (ADS)

    Weiss, Y.; Kiflawi, I.; Davies, N.; Navon, O.

    2014-09-01

    The chemical nature and composition of the growth medium of monocrystalline (MC) diamonds is still a matter of debate, partially because carbonate-bearing high-density fluids (HDFs) that are common in fibrous diamonds have not been found in MC diamonds. Here we report the first finding of HDF microinclusions in a MC octahedral diamond from Finsch, South Africa and in the MC octahedral core of a coated diamond from Kankan, Guinea; both diamonds carry nitrogen in B-centers. Numerous microinclusions in diamond Finsch_2a_cap1 are restricted to two thin layers parallel to the (1 1 1) face, ∼20 and 200 μm from the diamond rim. Low-Mg carbonatitic HDFs are found along the inner layer while the outer layer trapped saline compositions. The major and trace element compositions of the inclusions and their infrared spectra are highly similar to those of microinclusions found in fibrous diamonds. A few isolated microinclusions of saline compositions are scattered around a sulfide inclusion in the center of the octahedral core of diamond ON-KAN-383. This evidence for the involvement of oxidized fluids in the formation of MC diamonds adds to previous reports on the antiquity of HDFs in fibrous diamonds, the presence of carbonate and halide phases in inclusions in MC diamonds and the similarity of trace element pattern of a MC diamond to those of low-Mg carbonatitic HDF in fibrous diamonds. In addition, we show that the interaction of HDFs with depleted garnets can produce sinusoidal REE patterns which are one of the primary features of lherzolitic and harzburgitic garnet inclusions in MC diamonds. Together, these observations suggest that HDFs are involved in the formation of many types of diamonds from the Archaean to the Phanerozoic. HDFs are trapped in large quantities during rapid, fibrous growth, but must also be present during the growth of many MC diamonds.

  12. A screen of chemical modifications identifies position-specific modification by UNA to most potently reduce siRNA off-target effects

    PubMed Central

    Bramsen, Jesper B.; Pakula, Malgorzata M.; Hansen, Thomas B.; Bus, Claus; Langkjær, Niels; Odadzic, Dalibor; Smicius, Romualdas; Wengel, Suzy L.; Chattopadhyaya, Jyoti; Engels, Joachim W.; Herdewijn, Piet; Wengel, Jesper; Kjems, Jørgen

    2010-01-01

    Small interfering RNAs (siRNAs) are now established as the preferred tool to inhibit gene function in mammalian cells yet trigger unintended gene silencing due to their inherent miRNA-like behavior. Such off-target effects are primarily mediated by the sequence-specific interaction between the siRNA seed regions (position 2–8 of either siRNA strand counting from the 5′-end) and complementary sequences in the 3′UTR of (off-) targets. It was previously shown that chemical modification of siRNAs can reduce off-targeting but only very few modifications have been tested leaving more to be identified. Here we developed a luciferase reporter-based assay suitable to monitor siRNA off-targeting in a high throughput manner using stable cell lines. We investigated the impact of chemically modifying single nucleotide positions within the siRNA seed on siRNA function and off-targeting using 10 different types of chemical modifications, three different target sequences and three siRNA concentrations. We found several differently modified siRNAs to exercise reduced off-targeting yet incorporation of the strongly destabilizing unlocked nucleic acid (UNA) modification into position 7 of the siRNA most potently reduced off-targeting for all tested sequences. Notably, such position-specific destabilization of siRNA–target interactions did not significantly reduce siRNA potency and is therefore well suited for future siRNA designs especially for applications in vivo where siRNA concentrations, expectedly, will be low. PMID:20453030

  13. Selective gene silencing in activated leukocytes by targeting siRNAs to the integrin lymphocyte function-associated antigen-1

    PubMed Central

    Peer, Dan; Zhu, Pengcheng; Carman, Christopher V.; Lieberman, Judy; Shimaoka, Motomu

    2007-01-01

    Silencing gene expression by RNAi is a powerful method for exploring gene function and validating drug targets and potentially for therapy. Lymphocytes and other primary blood cells are resistant to lipid-based transfection in vitro and are difficult to target in vivo. We show here that antibody-protamine fusion proteins targeting the human integrin lymphocyte function-associated antigen-1 (LFA-1) efficiently deliver siRNAs and specifically induce silencing in primary lymphocytes, monocytes, and dendritic cells. Moreover, a fusion protein constructed from an antibody that preferentially recognizes activation-dependent conformational changes in LFA-1 selectively targets activated leukocytes and can be used to suppress gene expression and cell proliferation only in activated lymphocytes. The siRNA-fusion protein complexes do not cause lymphocyte activation or induce IFN responses. K562 cells expressing latent WT or constitutively activated LFA-1 engrafted in the lungs of SCID mice are selectively targeted by intravenously injected fusion protein–siRNA complexes, demonstrating the potential in vivo applicability of LFA-1-directed siRNA delivery. PMID:17360483

  14. Fe and Fe+2%Si targets as ion sources via UV laser ablation plasma

    NASA Astrophysics Data System (ADS)

    Lorusso, A.; Krása, J.; Láska, L.; Nassisi, V.; Velardi, L.

    2009-08-01

    In the last years the ion component of a laser-produced plasma has been considered and studied as an object to provide high-density ion sources, which can be applied in many fields such as laser-induced implantation. In this work a KrF laser beam of 108 W/cm2 irradiance was focused onto single-crystalline Fe and single-crystalline Fe with 2% of Si targets and the characteristics of both free expanding laser-produced plasmas were compared. The time-of-flight (TOF) method was applied to determine the ion charge yield at various laser fluences and the ion angular spread. The analyses of TOF spectra showed a synergetic effect of the silicon admixture in target material on the Fe ions production. Besides, this admixture was also responsible of the increasing of the plasma temperature which corresponds in turn to the increasing of the average kinetic energy of the particles as well as of the more collimated ion distribution.

  15. Nanoindentation-induced phase transformation and structural deformation of monocrystalline germanium: a molecular dynamics simulation investigation

    PubMed Central

    2013-01-01

    Molecular dynamics simulations were conducted to study the nanoindentation of monocrystalline germanium. The path of phase transformation and distribution of transformed region on different crystallographic orientations were investigated. The results indicate the anisotropic behavior of monocrystalline germanium. The nanoindentation-induced phase transformation from diamond cubic structure to β-tin-Ge was found in the subsurface region beneath the tool when indented on the (010) plane, while direct amorphization was observed in the region right under the indenter when the germanium was loaded along the [101] and [111] directions. The transformed phases extend along the < 110 > slip direction of germanium. The depth and shape of the deformed layers after unloading are quite different according to the crystal orientation of the indentation plane. The study results suggest that phase transformation is the dominant mechanism of deformation of monocrystalline germanium film in nanoindentation. PMID:23947487

  16. Long-term strength determination for cooled blades made of monocrystalline superalloys

    NASA Astrophysics Data System (ADS)

    Getsov, L. B.; Semenov, A. S.; Besschetnov, V. A.; Grishchenko, A. I.; Semenov, S. G.

    2017-04-01

    For the manufacture of blades for modern gas-turbine installations, monocrystalline alloys are used. Traditional methods for the calculation of stressed-deformed state and safety factors for these alloys developed and verified for polycrystalline materials need to be adjusted. This paper deals with methodological principles for an approach to the solving of the problem concerning a finite-element determination of the long-term static strength for cooled monocrystalline blades employed in gas-turbine installations based on the use of two different models (phenomenological and micromechanical) considering the inelastic deformation of monocrystalline superalloys. An analysis has been performed for the distribution of Schmid factors in the spherical triangle for primary and secondary octahedral and cubic slip systems. Calculations are performed using Larson-Miller's parametric dependences taking into account the crystallographic orientation of the material. A determination procedure for the anisotropy coefficients of long-term strength is described based on data for different orientations. A comparative analysis of the results of finite-element calculations made using phenomenological and micromechanical (crystallographic) creep models for the long-term static strength of cooled monocrystalline blades used in a gas-turbine engine has been performed. It is shown that the location of the most loaded sections of such a blade coincide with the results of calculations according to these models. It has been found that the micromechanical deformation model results in the obtaining of the most conservative estimate for the long-term strength of turbine blades made of monocrystalline alloys. It is shown that the calculations using models for materials with isotropic properties can produce considerable errors in determining the durability of the blades. The possibility is considered for using 1D-, 2D-, and 3D-models for turbine monocrystalline blades in the determination of

  17. Fabrication of Sr silicate buffer layer on Si(100) substrate by pulsed laser deposition using a SrO target

    SciTech Connect

    Imanaka, Atsuhiro; Sasaki, Tsubasa; Hotta, Yasushi Satoh, Shin-ichi

    2014-09-01

    The authors fabricated 2 × 1 Sr-reconstructed Si(100) substrates using thin SrO layers, and used them to direct growth of crystalline perovskite oxide on Si. The SrO layers used to reconstruct the Si(100) substrates were grown by pulsed laser deposition from a SrO single crystal target, followed by postdeposition-annealing (PDA) of the SrO/Si(100) structure. In situ observations of reflective high-energy electron diffraction during PDA confirmed a 2 × 1 reconstruction of the Si surface and x-ray photoemission spectroscopy of the annealed samples confirmed the existence of Sr atoms in a silicate phase, which indicated that a 2 × 1 Sr-reconstructed Si surface was achieved. The optimal fabrication conditions were annealing at 720 °C for 1 min and an equivalent SrO layer thickness (ML{sub eq}) of 2.5 ML{sub eq}. The temperature condition was very narrow, at 720 ± 20 °C, for an acceptable product. Subsequently, the authors demonstrated the growth of crystalline SrTiO{sub 3} films on the 2 × 1 Sr-reconstructed Si(100) surfaces.

  18. Cytosolic mRNA Target and Bioavailability of Nanoparticulate siRNA delivery systems for gene silencing.

    PubMed

    Leucuta, Sorin Emilian

    2017-03-22

    Recent research in medical and pharmaceutical sciences has benefited from advances in molecular biology and genetics, which made possible a diagnosis at the molecular level in more and more diseases. This implies the drug treatment at the molecular level. The interest in Ribonucleic acid interference (RNAi) is based on the mechanism operates by eliminating the messenger RNAs (mRNAs) coding for multiple proteins, which open solutions for treating many types of diseases. Small (short) interfering RNA (siRNA) has quickly been established as an effective gene-silencing strategy in animal models, and more recently in human clinical trials, as a potential therapeutic approach. Various nanoparticulate drug delivery systems for siRNA delivery have been explored extensively. However, there are many more barriers and challenges that need to be addressed and overcome to achieve the ideal formulation in terms of selectivity, efficacy and safety. One of the major causes of the drawback of these treatments is the difficulty to transport the nucleic acids in the cytosol and organelles. These delivery systems will favorably alter the pharmacokinetics and biodistribution of siRNAs, should be biocompatible and genocompatible to avoid immune stimulation and off-target gene effects. These properties are essential for systemic use, as they prolong siRNA half-lives in blood and increase intracellular bioavailability of siRNA. Future research needs drug delivery systems with more effective design, enhanced biological stability, subcellular bioavailability, and efficient targeted delivery in vivo for improved targeting and specificity of siRNA molecules for any given clinical condition. The paper shows how to overcome physiological barriers to achieve the target, and examples in which significant results were obtained in therapeutic in vitro and in vivo research including nanoparticulate systems.To day, only a few nanoparticle-based siRNA delivery systems have been approved by the Food

  19. Trilayer micelles for combination delivery of rapamycin and siRNA targeting Y-box binding protein-1 (siYB-1)

    PubMed Central

    Zeng, San; Xiong, May P.

    2013-01-01

    A three layer (trilayer) polymeric micelle system based on the self-association of the triblock polymer poly(ethylene glycol)-b-poly{N-[N-(2-aminoethyl)-2-aminoethyl] aspartamide}-b-poly(ε-caprolactone) (PEG-b-PAsp(DET)-b-PCL) has been synthesized and investigated for combination delivery of rapamycin (RAP) and siRNA targeting Y-box binding protein-1 (siYB-1). The trilayer micelle is composed of (a) a hydrophilic poly(ethylene glycol) (PEG) block constituting the outer layer to improve pharmacokinetics, (b) an intermediate compartment composed of the cationic poly{2-[(2-aminoethyl)amino] ethyl aspartamide} (PAsp(DET)) segment for interacting with siYB-1, and (c) an inner hydrophobic poly(ε-caprolactone) (PCL) compartment for encapsulation of RAP. A major advantage of this system is biocompatibility since PEG and PCL are both approved by the FDA, and PAsp(DET) is a non-toxic pH responsive cationic poly(amino acid)-based polymer. In this study, it has been shown that PCL can encapsulate RAP with high loading efficiencies, and PAsp(DET) can successfully interact with siRNA for efficient transfection/knockdown with negligible cytotoxicity. The enhanced therapeutic efficacy of RAP/ siYB-1 micelles was demonstrated in cell cultures and in a PC3 xenograft nude mouse model of human prostate cancer. Herein, we demonstrate that trilayer micelles are a promising approach to improve the simultaneous delivery of combination siRNA/drug therapies. PMID:23768780

  20. siRNA as a tool to improve the treatment of brain diseases: Mechanism, targets and delivery.

    PubMed

    Gomes, Maria João; Martins, Susana; Sarmento, Bruno

    2015-05-01

    As the population ages, brain pathologies such as neurodegenerative diseases and brain cancer increase their incidence, being the need to find successful treatments of upmost importance. Drug delivery to the central nervous system (CNS) is required in order to reach diseases causes and treat them. However, biological barriers, mainly blood-brain barrier (BBB), are the key obstacles that prevent the effectiveness of possible treatments due to their ability to strongly limit the perfusion of compounds into the brain. Over the past decades, new approaches towards overcoming BBB and its efflux transporters had been proposed. One of these approaches here reviewed is through small interfering RNA (siRNA), which is capable to specifically target one gene and silence it in a post-transcriptional way. There are different possible functional proteins at the BBB, as the ones responsible for transport or just for its tightness, which could be a siRNA target. As important as the effective silence is the way to delivery siRNA to its anatomical site of action. This is where nanotechnology-based systems may help, by protecting siRNA circulation and providing cell/tissue-targeting and intracellular siRNA delivery. After an initial overview on incidence of brain diseases and basic features of the CNS, BBB and its efflux pumps, this review focuses on recent strategies to reach brain based on siRNA, and how to specifically target these approaches in order to treat brain diseases. Copyright © 2015 Elsevier B.V. All rights reserved.

  1. Identification of novel cellular targets for therapeutic intervention against Ebola virus infection by siRNA screening.

    PubMed

    Kolokoltsov, Andrey A; Saeed, Mohammad F; Freiberg, Alexander N; Holbrook, Michael R; Davey, Robert A

    2009-06-01

    While much progress has been made in developing drugs against a few prominent viruses such as HIV, few examples exist for emerging infectious agents. In some cases broad spectrum anti-viral drugs, such as ribavirin, are effective, but for some groups of viruses, these show little efficacy in animal models. Traditional methods focus on screening small molecule libraries to identify drugs that target virus factors, with the intention that side-effects to the host can be minimized. However, this greatly limits potential drug targets and virus genes can rapidly mutate to avoid drug action. Recent advances in siRNA gene targeting technologies have provided a powerful tool to specifically target and suppress the expression of cell genes. Since viruses are completely dependent upon host cell proteins for propagation, siRNA screening promises to reveal novel cell proteins and signaling pathways that may be viable targets for drug therapy regimens. Here we used an siRNA screening approach to identify gene products that play critical roles in Ebola virus infection. By gene cluster analysis, proteins in phosphatidylinositol-3-kinase and calcium/calmodulin kinase related networks were identified as important for Zaire Ebola virus infection and prioritized for further evaluation. Key roles of each were confirmed by testing available drugs specific for members of each pathway. Interestingly, both sets of proteins are also important in cancer and subject to intense investigation. Thus development of new drugs against these cancer targets may also prove useful in combating Ebola virus.

  2. Preparation and evaluation of nanoparticles loading plasmid DNAs inserted with siRNA fragments targeting c-Myc gene.

    PubMed

    Ma, Tao; Jiang, Jin-Ling; Liu, Ying; Ye, Zheng-Bao; Zhang, Jun

    2014-09-01

    c-Myc plays a key role in glioma cancer stem cell maintenance. A drug delivery system, nanoparticles loading plasmid DNAs inserted with siRNA fragments targeting c-Myc gene (NPs-c-Myc-siRNA-pDNAs), for the treatment of glioma, has not previously been reported. NPs-c-Myc-siRNA-pDNAs were prepared and evaluated in vitro. Three kinds of c-Myc-siRNA fragments were separately synthesized and linked with empty siRNA expression vectors in the mole ratio of 3:1 by T4 DNA ligase. The linked products were then separately transfected into Escherichia coli. DH5α followed by extraction with Endofree plasmid Mega kit (Qiagen, Hilden, Germany) obtained c-Myc-siRNA-pDNAs. Finally, the recombinant c-Myc-siRNA3-pDNAs, generating the highest transfection efficiency and the greatest apoptotic ability, were chosen for encapsulation into NPs by the double-emulsion solvent-evaporation procedure, followed by stability, transfection efficiency, as well as qualitative and quantitative apoptosis evaluation. NPs-c-Myc-siRNA3-pDNAs were obtained with spherical shape in uniform size below 150 nm, with the zeta potential about -18 mV, the encapsulation efficiency and loading capacity as 76.3 ± 5.4% and 1.91 ± 0.06%, respectively. The stability results showed that c-Myc-siRNA3-pDNAs remained structurally and functionally stable after encapsulated into NPs, and NPs could prevent the loaded c-Myc-siRNA3-pDNAs from DNase degradation. The transfection efficiency of NPs-c-Myc-siRNA3-pDNAs was proven to be positive. Furthermore, NPs-c-Myc-siRNA3-pDNAs produced significant apoptosis with the apoptotic rate at 24.77 ± 5.39% and early apoptosis cells observed. Methoxy-poly-(ethylene-glycol)-poly-(lactide-co-glycolide) nanoparticles (MPEG-PLGA-NPs) are potential delivery carriers for c-Myc-siRNA3-pDNAs.

  3. A hybrid siRNA delivery complex for enhanced brain penetration and precise amyloid plaque targeting in Alzheimer's disease mice.

    PubMed

    Zheng, Xiaoyao; Pang, Xiaoying; Yang, Peng; Wan, Xu; Wei, Yue; Guo, Qian; Zhang, Qizhi; Jiang, Xinguo

    2017-02-01

    To realize the therapeutic potential of gene drugs for Alzheimer's disease (AD), non-invasive, tissue-specific and efficient delivery technologies must be developed. Here, a hybrid system for amyloid plaques targeted siRNA delivery was formed by PEGylated Poly(2-(N,N-dimethylamino) ethyl methacrylate) (PEG-PDMAEMA) conjugated with two d-peptides, a CGN for brain penetration and a QSH for β-amyloid binding. The hybrid complex CQ/siRNA, composed of 25% MPEG-PDMAEMA, 50% CGN-PEG-PDMAEMA and 25% QSH-PEG-PDMAEMA, showed negligible cytotoxicity and could protect siRNA from enzyme degradation. Being taken up by neuron cells, the complexes could escape from lysosomes, release siRNA in the cytoplasm and thus producing effective gene silence (down-regulated protein level to 18.5%). After intravenous injection, CQ/siRNA penetrated into the brain in an intact form and located around the plaques in transgenic AD mice. The precisely amyloid plaques delivery resulted in increased therapeutic activities, which was demonstrated by the strong mRNA (36.4%) knockdown of BACE1 (a therapeutic target of AD), the less yield of enzyme-digested products sAPPβ (-42.6%), as well as the better neurons protection than the single component complexes. In conclusion, the hybrid complex could efficiently and precisely deliver an siRNA to the AD lesion and might be a potential candidate for gene therapy for AD. The gene delivery system achieving high brain penetration and lesion region accumulation was first applied to treat AD, and the preparation exhibited a significantly better neuroprotective effect than that modified with a single ligand. The intracellular process of which the complexes escape from lysosomes and release the siRNA in cytoplasm was revealed. The brain targeting and amyloid plaque binding ability of the complex were systemic evaluated, and the in vivo co-location experiments provided a direct evidence of the precise delivery of the siRNA to the amyloid plaques. One of the

  4. Tumor responsive targeted multifunctional nanosystems for cancer imaging, chemo- and siRNA therapy

    NASA Astrophysics Data System (ADS)

    Savla, Ronak

    Cancer is one of the most insidious diseases. Compromising of over 100 different types and sharing the unifying factors of uncontrolled growth and metastasis, unmet clinical needs in terms of cancer diagnosis and treatment continue to exist. It is widely accepted that most forms of cancer are treatable or even curable if detected before widespread metastasis occurs. Nearly a quarter of deaths in the United States is the result of cancer and it only trails heart disease in terms of annual mortality. Surgery, chemotherapy, and radiation therapy are the primary treatment modalities for cancer. Research in these procedures has resulted in substantial benefits for cancer patients, but there is still room for an improvement. However, a time has been reached at which it appears that the benefits from these modalities have been reached the maximum. Therefore, it is vital to develop new strategies for the diagnosis and treatment of cancer. The field of nanotechnology is concerned with structures in the nanometer size range and holds the potential to drastically impact and improve the lives of patients suffering from cancer. Not only can nanotechnology improve current methods of diagnosis and treatment, it has a possibility of introducing newer and better modalities. The overall purpose of this work is to develop novel nanotechnology-based methodologies for the diagnosis and treatment of various forms of cancers. The first aim of the project is the development of a multifunctional targeted nanosystem for the delivery of siRNA to overcome drug resistance. The second aspect is the synthesis of a quantum dot-based delivery system that releases drug in response to pH changes. The third aim is the development of a targeted, tumor environment responsive magnetic resonance nanoparticle contrast agent coupled with a nanoparticle-based treatment.

  5. LncRNA260-specific siRNA targeting IL28RA gene inhibit cardiomyocytes hypoxic/reoxygenation injury.

    PubMed

    Gong, Ge; Yang, Xin-Xing; Li, Yanyan; Geng, Hong-Yu; Yang, Zhi-Jian; Wang, Lian-Sheng; Kim, Hyun Jun; Lu, Xin-Zheng

    2017-08-01

    The interleukin 28 receptor alpha (IL28RA) gene was indicated to be associated with apoptosis. However, it was not clear whether long non-coding RNA 260 (lncRNA 260)-specific siRNA targeting IL28RA gene could inhibit hypoxic reoxygenation (H/R) cardiomyocytes injury or not. To explore the mechanisms underlying the protective effects of lncRNA260-specific siRNA-mediated inhibition of IL28RA from H/R injury in cardiomyocytes, the current research was performed. The primary neonatal rat cardiomyocytes were transfected with three different pairs of siRNA specific to lncRNA260 targeting IL28RA gene and then were undergone with the conditions simulating H/R injury. All three groups of cardiomyocytes treated with lncRNA260-specific siRNA experienced significantly decreased levels of lactate dehydrogenase activity and apoptosis rate relative to the non-treatment and negative control groups (P<0.05), also expressed reduced levels of IL28RA, and increased levels of PI3KCG and Bcl-2/Bax (P<0.05). The lncRNA260-specific siRNA may reduce cardiomyocyte apoptosis associated with H/R injury by decreasing levels of the IL28RA gene product and thus activating the PI3K/AKT signaling pathway.

  6. Single base mismatches in the mRNA target site allow specific seed region-mediated off-target binding of siRNA targeting human coagulation factor 7.

    PubMed

    Ravon, Morgane; Berrera, Marco; Ebeling, Martin; Certa, Ulrich

    2012-01-01

    We have analyzed the off-target activity of two siRNAs (F7-1, F7-2) that knock-down human blood coagulation factor 7 mRNA. F7-1 modulates a significant number of non-target transcripts while F7-2 shows high selectivity for the target transcript under various experimental conditions. The 3'-UTRs of all F7-1 off-target genes show statistically significant enrichment of the reverse complement of the F7-1 siRNA seed region located in the guide strand. Seed region enrichment was confirmed in off-target transcripts modulated by siRNA targeting the glucocorticoid receptor. To investigate how these sites contribute to off-target recognition of F7-1, we employed CXCL5 transcript as model system because it contains five F7-1 seed sequence motifs with single base mismatches. We show by transient transfection of reporter gene constructs into HEK293 cells that three out of five sites located in the 3'-UTR region are required for F7-1 off-target activity. For further mechanistic dissection, the sequences of these sites were synthesized and inserted either individually or joined in dimeric or trimeric constructs. Only the fusion constructs were silenced by F7-1 while the individual sites had no off-target activity. Based on F7-1 as a model, a single mismatch between the siRNA seed region and mRNA target sites is tolerated for target recognition and the CXCL5 data suggest a requirement for binding to multiple target sites in off-target transcripts.

  7. Single base mismatches in the mRNA target site allow specific seed region-mediated off-target binding of siRNA targeting human coagulation factor 7

    PubMed Central

    Ravon, Morgane; Berrera, Marco; Ebeling, Martin; Certa, Ulrich

    2012-01-01

    We have analyzed the off-target activity of two siRNAs (F7-1, F7-2) that knock-down human blood coagulation factor 7 mRNA. F7-1 modulates a significant number of non-target transcripts while F7-2 shows high selectivity for the target transcript under various experimental conditions. The 3′-UTRs of all F7-1 off-target genes show statistically significant enrichment of the reverse complement of the F7-1 siRNA seed region located in the guide strand. Seed region enrichment was confirmed in off-target transcripts modulated by siRNA targeting the glucocorticoid receptor. To investigate how these sites contribute to off-target recognition of F7-1, we employed CXCL5 transcript as model system because it contains five F7-1 seed sequence motifs with single base mismatches. We show by transient transfection of reporter gene constructs into HEK293 cells that three out of five sites located in the 3′-UTR region are required for F7-1 off-target activity. For further mechanistic dissection, the sequences of these sites were synthesized and inserted either individually or joined in dimeric or trimeric constructs. Only the fusion constructs were silenced by F7-1 while the individual sites had no off-target activity. Based on F7-1 as a model, a single mismatch between the siRNA seed region and mRNA target sites is tolerated for target recognition and the CXCL5 data suggest a requirement for binding to multiple target sites in off-target transcripts. PMID:22258146

  8. siRNA targeting of Trop2 suppresses the proliferation and invasion of lung adenocarcinoma H460 cells.

    PubMed

    Gao, Xiao-Yan; Zhu, Ye-Han; Zhang, Li-Xin; Lu, Hui-Yu; Jiang, Ai-Gui

    2015-08-01

    The aim of the present study was to investigate the effect of the small interfering RNA (siRNA)-induced inhibition of the Trop2 gene on the proliferation and invasion of lung adenocarcinoma H460 cells. A recombinant adenovirus expression vector, which contained siRNA targeting open reading frames for Trop2 (rAd5-siTrop2), was transfected into lung adenocarcinoma H460 cells. Three groups were included in the study, namely the Ctrl (non-transfected control), rAd5-siCtrl (native control) and rAd5-siTrop2 (knockdown Trop2 gene) groups. The mRNA and protein expression levels of Trop2 were detected using quantitative polymerase chain reaction and western blot analysis, respectively. In addition, the expression levels of cyclin Dl and phospho-extracellular signal regulated kinase (p-ERK)-1 were detected using western blot analysis. The effects of Trop2 inhibition on the proliferation and invasion of lung adenocarcinoma H460 cells were investigated using a 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide and Transwell assay. Trop2-targeted siRNA recombinant plasmids were successfully constructed. The recombinant adenovirus vector, rAd5-siTrop2, significantly downregulated the mRNA and protein expression levels of Trop2 in the lung adenocarcinoma H460 cells, with cyclin D1 and p-ERK-1 expression downregulated simultaneously. In addition, following the silencing of Trop2, the proliferation and invasion rates of the lung adenocarcinoma H460 cells were reduced. Therefore, the results indicated that Trop2 serves a key function in the proliferation and invasion of lung adenocarcinoma H460 cells in vitro.

  9. The Effects of Cells Temperature Increment and Variations of Irradiation for Monocrystalline Photovoltaic

    NASA Astrophysics Data System (ADS)

    Fuad Rahman Soeharto, Faishal; Hermawan

    2017-04-01

    Photovoltaic cell technology has been developed to meet the target of 17% Renewable Energy in 2025 accordance with Indonesia Government Regulation No. 5 2006. Photovoltaic cells are made of semiconductor materials, namely silicon or germanium (p-n junction). These cells need the light that comes from solar irradiation which brings energy photons to convert light energy into electrical energy. It is different from the solar heater that requires heat energy or thermal of sunlight that is normally used for drying or heating water. Photovoltaic cells requires energy photons to perform the energy conversion process, the photon energy can be derived from sunlight. Energy photon is taken from the sun light along with the advent of heat due to black-body radiation, which can lead to temperature increments of photovoltaic cells. Increment of 1°C can decreased photovoltaic cell voltage of up to 2.3 mV per cell. In this research, it will be discuss the analysis of the effect of rising temperatures and variations of irradiation on the type monocrystalline photovoltaic. Those variation are analyzed, simulated and experiment by using a module of experiment. The test results show that increment temperature from 25° C to 80° C at cell of photovoltaic decrease the output voltage of the photovoltaic cell at 4.21 V, and it also affect the power output of the cell which decreases up to 0.7523 Watt. In addition, the bigger the value of irradiation received by cell at amount of 1000 W / m2, produce more output power cells at the same temperature.

  10. pH-sensitive siRNA Nanovector for Targeted Gene Silencing and Cytotoxic Effect in Cancer Cells

    PubMed Central

    Mok, Hyejung; Veiseh, Omid; Fang, Chen; Kievit, Forrest M.; Wang, Freddy Y.; Park, James O.; Zhang, Miqin

    2010-01-01

    A small interfering RNA (siRNA) nanovector with dual targeting specificity and dual therapeutic effect is developed for targeted cancer imaging and therapy. The nanovector is comprised of an iron oxide magnetic nanoparticle core coated with three different functional molecules: polyethyleneimine (PEI), siRNA, and chlorotoxin (CTX). The primary amine group of PEI is blocked with citraconic anhydride that is removable at acidic conditions, not only to increase its biocompatibility at physiological conditions but also to elicit a pH-sensitive cytotoxic effect in the acidic tumor microenvironment. The PEI is covalently immobilized on the nanovector via a disulfide linkage that is cleavable after cellular internalization of the nanovector. CTX as a tumor-specific targeting ligand and siRNA as a therapeutic payload are conjugated on the nanovector via a flexible and hydrophilic PEG linker for targeted gene silencing in cancer cells. With a size of ~ 60 nm, the nanovector exhibits long-term stability and good magnetic property for magnetic resonance imaging. The multifunctional nanovector exhibits both significant cytotoxic and gene silencing effects at acidic pH conditions for C6 glioma cells, but not at physiological pH conditions. Our results suggest that this nanovector system could be safely used as a potential therapeutic agent for targeted treatment of glioma as well as other cancers. PMID:20722417

  11. Targeted exosome-mediated delivery of opioid receptor Mu siRNA for the treatment of morphine relapse

    PubMed Central

    Liu, Yuchen; Li, Dameng; Liu, Zhengya; Zhou, Yu; Chu, Danping; Li, Xihan; Jiang, Xiaohong; Hou, Dongxia; Chen, Xi; Chen, Yuda; Yang, Zhanzhao; Jin, Ling; Jiang, Waner; Tian, Chenfei; Zhou, Geyu; Zen, Ke; Zhang, Junfeng; Zhang, Yujing; Li, Jing; Zhang, Chen-Yu

    2015-01-01

    Cell-derived exosomes have been demonstrated to be efficient carriers of small RNAs to neighbouring or distant cells, highlighting the preponderance of exosomes as carriers for gene therapy over other artificial delivery tools. In the present study, we employed modified exosomes expressing the neuron-specific rabies viral glycoprotein (RVG) peptide on the membrane surface to deliver opioid receptor mu (MOR) siRNA into the brain to treat morphine addiction. We found that MOR siRNA could be efficiently packaged into RVG exosomes and was associated with argonaute 2 (AGO2) in exosomes. These exosomes efficiently and specifically delivered MOR siRNA into Neuro2A cells and the mouse brain. Functionally, siRNA-loaded RVG exosomes significantly reduced MOR mRNA and protein levels. Surprisingly, MOR siRNA delivered by the RVG exosomes strongly inhibited morphine relapse via the down-regulation of MOR expression levels. In conclusion, our results demonstrate that targeted RVG exosomes can efficiently transfer siRNA to the central nervous system and mediate the treatment of morphine relapse by down-regulating MOR expression levels. Our study provides a brand new strategy to treat drug relapse and diseases of the central nervous system. PMID:26633001

  12. Analysis of the siRNA-Mediated Gene Silencing Process Targeting Three Homologous Genes Controlling Soybean Seed Oil Quality.

    PubMed

    Lu, Sha; Yin, Xiaoyan; Spollen, William; Zhang, Ning; Xu, Dong; Schoelz, James; Bilyeu, Kristin; Zhang, Zhanyuan J

    2015-01-01

    In the past decade, RNA silencing has gained significant attention because of its success in genomic scale research and also in the genetic improvement of crop plants. However, little is known about the molecular basis of siRNA processing in association with its target transcript. To reveal this process for improving hpRNA-mediated gene silencing in crop plants, the soybean GmFAD3 gene family was chosen as a test model. We analyzed RNAi mutant soybean lines in which three members of the GmFAD3 gene family were silenced. The silencing levels of FAD3A, FAD3B and FAD3C were correlated with the degrees of sequence homology between the inverted repeat of hpRNA and the GmFAD3 transcripts in the RNAi lines. Strikingly, transgenes in two of the three RNAi lines were heavily methylated, leading to a dramatic reduction of hpRNA-derived siRNAs. Small RNAs corresponding to the loop portion of the hairpin transcript were detected while much lower levels of siRNAs were found outside of the target region. siRNAs generated from the 318-bp inverted repeat were found to be diced much more frequently at stem sequences close to the loop and associated with the inferred cleavage sites on the target transcripts, manifesting "hot spots". The top candidate hpRNA-derived siRNA share certain sequence features with mature miRNA. This is the first comprehensive and detailed study revealing the siRNA-mediated gene silencing mechanism in crop plants using gene family GmFAD3 as a test model.

  13. Synthetic siRNAs effectively target cystein protease 12 and α-actinin transcripts in Trichomonas vaginalis.

    PubMed

    Ravaee, Roya; Ebadi, Parimah; Hatam, Gholamreza; Vafafar, Arghavan; Ghahramani Seno, Mohammad Mahdi

    2015-10-01

    The flagellated protozoan Trichomonas vaginalis (T. vaginalis) causes trichomoniasis, a reproductive tract infection, in humans. Trichomoniasis is the most common non-viral sexually transmitted disease worldwide. In addition to direct consequences such as infertility and abortion, there are indications that trichomoniasis favours development of prostate cancer and it has also been associated with increased risk of spreading human immunodeficiency virus and papillomavirus infections. Reports from around the world show that the rate of drug resistance in T. vaginalis is increasing, and therefore new therapeutic approaches have to be developed. Studying molecular biology of T. vaginalis will be quite helpful in identifying new drugable targets. RNAi is a powerful technique which allows biologist to specifically target gene products (i.e. mRNA) helping them in unravelling gene functions and biology of systems. However, due to lack of some parts of the required intrinsic RNAi machinery, the RNAi system is not functional in all orders of life. Here, by using synthetic siRNAs targeting two genes, i.e. α-actinin and cystein protease 12 (cp12), we demonstrate T. vaginalis cells are amenable to RNAi experiments conducted by extrinsic siRNAs. Electroporation of siRNAs targeting α-actinin or cp12 into T. vaginalis cells resulted in, respectively, 48-67% and 33-72% downregulation of the cognate transcripts compared to the T. vaginalis cells received siRNAs targeting GL2 luciferase as a control. This finding is helpful in that it demonstrates the potential of using extrinsically induced RNAi in studies on molecular biology of T. vaginalis such as those aiming at identifying new drug targets. Copyright © 2015 Elsevier Inc. All rights reserved.

  14. Targeted in vivo delivery of EGFR siRNA inhibits ovarian cancer growth and enhances drug sensitivity

    PubMed Central

    Satpathy, Minati; Mezencev, Roman; Wang, Lijuan; McDonald, John F.

    2016-01-01

    A functionalized nanohydrogel siRNA delivery system and a mouse model of serous ovarian cancer were used to test predictions from previous cell line studies that knockdown of EGFR (epidermal growth factor receptor) may be of clinical significance in the treatment of epithelial tumors especially with respect to the enhancement of platinum based therapies. Our results support these predictions and suggest that targeted delivery of EGFR siRNA may be an effective strategy for the treatment of ovarian and other epithelial tumors associated with elevated levels of EGFR and especially those demonstrating resistance to platinum-based therapies. PMID:27819259

  15. Functionalized silicon quantum dots tailored for targeted siRNA delivery

    SciTech Connect

    Klein, S.; Zolk, O.; Fromm, M.F.; Schroedl, F.; Kryschi, C.

    2009-09-11

    For RNA interference (RNAi) mediated silencing of the ABCB1 gene in Caco-2 cells biocompatible luminescent silicon quantum dots (SiQDs) were developed to serve as self-tracking transfection tool for ABCB1 siRNA. While the 2-3 nm sized SiQD core exhibits green luminescence, the QD surfaces are completely saturated with covalently linked 2-vinylpyridine that may electrostatically bind siRNA. For down-regulating P-glycoprotein (Pgp) expression of the ABCB1 gene the SiQDs were complexed with siRNA. The cellular uptake and allocation of SiQD-siRNA complexes in Caco-2 cells were monitored using confocal laser scanning microscopy and transmission electron microscopy. The release of siRNA to the cytoplasm was verified through real-time PCR quantification of the reduced ABCB1 mRNA level. Additional evidence was obtained from time-resolved in situ fluorescence spectroscopic monitoring of the Pgp efflux dynamics in transfected Caco-2 cells which yielded significantly reduced transporter efficiencies for the Pgp substrate Rhodamine 123.

  16. In situ hole doping of wide-gap semiconductors by dual-target simultaneous laser ablation: GaN and SiC epitaxial films

    NASA Astrophysics Data System (ADS)

    Muto, Hachizo; Asano, Takashi; Wang, Rong-Ping; Kusumori, Takeshi

    2005-10-01

    Apparatus for dual-target simultaneous laser ablation deposition and in situ doping techniques have been developed to achieve p-type doping during epitaxial growth of wide-band-gap semiconductors. The apparatus has two target holders with a target-rotation mechanism and a rotation-axis adjusting mechanism to obtain homogeneously doped films. Mg-doped GaN films have been fabricated on 6H-SiC(0001) and Si(111) substrates in NH3 ambient by simultaneous ablation of GaN and Mg-metal targets using two lasers. Junctions of the films with n-type substrates show a diode curve characteristic of p-n junctions, but not for junction with p-Si, indicating hole doping without further procedures. In situ p-type doping to SiC was also achieved by using SiC and Al4C3 targets.

  17. SiRNA Targeting mTOR Effectively Prevents the Proliferation and Migration of Human Lens Epithelial Cells

    PubMed Central

    Zhang, Chunmei; Liu, Jingjing; Jin, Na; Zhang, Guiming; Xi, Yahui; Liu, Hongling

    2016-01-01

    Posterior capsule opacification (PCO) is the most common complication that causes visual decrease after extracapsular cataract surgery. The primary cause of PCO formation is the proliferation of the residual lens epithelial cells (LECs). The mammalian target of rapamycin (mTOR) plays an important role in the growth and migration of LECs. In the current study, we used small interfering RNA (siRNA) to specifically attenuate mTOR in human lens epithelial B3 cells (HLE B3). We aimed to examine the effect of mTOR-siRNA on the proliferation, migration and epithelial-to-mesenchymal transition (EMT) of HLE B3 cells and explore the underlying mechanisms. The mTOR-siRNA was transfected into HLE B3 cells using lipofectamine 2000. The mRNA and protein levels of mTOR were examined to confirm the efficiency of mTOR-siRNA. The levels of mRNA and protein as well as the activity of mTOR down-stream effectors p70 ribosomal protein S6 kinase (p70S6K) and protein kinase B (PKB, AKT) were examined using real-time PCR or Western blot, respectively. The cell proliferation was determined using cell counting kit (CCK) 8 and cell growth curve assay. The cell migration was examined using Transwell system and Scratch assay. MTOR-siRNA effectively eliminated mTOR mRNA and protein. The proliferation and migration were significantly suppressed by mTOR-siRNA transfection. mTOR-siRNA reduced the mRNA of p70S6K and AKT in a time-dependent manner. Furthermore, the phosphorylation of p70S6K and AKT was decreased by mTOR-siRNA. MTOR-siRNA also eliminated the formation of mTORC1 and mTORC2 protein complex and blocked the transforming growth factor (TGF)-β-induced EMT. Our results suggested that mTOR-siRNA could effectively inhibit the proliferation, migration and EMT of HLE B3 cells through the inhibition of p70S6K and AKT. These results indicated that mTOR-siRNA might be an effective agent inhibiting HLE cells growth and EMT following cataract surgery and provide an alternative therapy for preventing

  18. Targeted delivery of biodegradable nanoparticles with ultrasound-targeted microbubble destruction-mediated hVEGF-siRNA transfection in human PC-3 cells in vitro.

    PubMed

    Li, Yun-Hua; Shi, Qiu-Sheng; Du, Jing; Jin, Li-Fang; Du, Lian-Fang; Liu, Pei-Feng; Duan, You-Rong

    2013-01-01

    A potentially viable approach for treating late-stage prostate cancer is gene therapy. Successful gene therapy requires safe and efficient delivery systems. In this study, we report the efficient delivery of small interfering RNA (siRNA) via the use of biodegradable nanoparticles (NPs) made from monomethoxypoly(ethylene glycol)-poly(lactic-co-glycolic acid)-poly-l-lysine (mPEG-PLGA-PLL) triblock copolymers. On the basis of previous findings, cyclic Arg-Gly-Asp (cRGD) peptides were conjugated to NPs to recognize the target site, integrin αvβ3, expressed in high levels in PC-3 prostate cancer cells. The suppression of angiogenesis by the downregulation of vascular endothelial growth factor (VEGF) expression has been widely used to inhibit the growth of malignant tumors. In our study, human VEGF (hVEGF)-siRNA was encapsulated in NPs to inhibit VEGF expression in PC-3 cells. Concurrently, sonoporation induced by ultrasound-targeted microbubble destruction (UTMD) was utilized for the delivery of siRNA-loaded NPs. Our results showed low cytotoxicity and high gene transfection efficiency, demonstrating that the targeted delivery of biodegradable NPs with UTMD may be potentially applied as new vector system for gene delivery.

  19. Human CD64-targeted non-viral siRNA delivery system for blood monocyte gene modulation

    PubMed Central

    Yong, Seok-Beom; Kim, Hyung Jin; Kim, Jang Kyoung; Chung, Jee Young; Kim, Yong-Hee

    2017-01-01

    A subset of phagocytes including inflammatory monocytes in blood migrate and give rise to macrophages in inflammatory tissues which generated the idea that blood monocytes are the therapeutic targets for drug delivery. Fc gamma receptor I (CD64) is a membrane receptor for the Fc region of immunoglobulin G, primarily expressed on monocyte-lineage, and H22 a monoclonal antibody for human CD64 had shown rapid blood monocyte binding and occupation in clinical studies. Small interfering RNA-mediated gene silencing as a therapeutic has been proposed and is a promising strategy in terms of its “knock-down” ability on the target gene prior to translation. However, its instability and off-targeting effect must be overcome for success in clinical studies. In this study, we developed a non-viral delivery system composed of oligo-nona-arginine (9R) and anti-human CD64 single chain antibodies (H22) for human monocyte-specific siRNA delivery. A targeted and efficient siRNA delivery mediated by anti-CD64 scFv-9R was observed in CD64 positive human leukemia cells, THP-1. With primary human blood cells, anti-CD64 scFv-9R mediated gene silencing was quantitatively confirmed representing blood monocyte selective gene delivery. These results demonstrate the potential of anti-CD64 scFv-9R mediated siRNA delivery for the treatment of human inflammatory diseases via blood monocytes gene delivery. PMID:28169353

  20. Long-circulating siRNA nanoparticles for validating Prohibitin1-targeted non-small cell lung cancer treatment.

    PubMed

    Zhu, Xi; Xu, Yingjie; Solis, Luisa M; Tao, Wei; Wang, Liangzhe; Behrens, Carmen; Xu, Xiaoyang; Zhao, Lili; Liu, Danny; Wu, Jun; Zhang, Ning; Wistuba, Ignacio I; Farokhzad, Omid C; Zetter, Bruce R; Shi, Jinjun

    2015-06-23

    RNA interference (RNAi) represents a promising strategy for identification and validation of putative therapeutic targets and for treatment of a myriad of important human diseases including cancer. However, the effective systemic in vivo delivery of small interfering RNA (siRNA) to tumors remains a formidable challenge. Using a robust self-assembly strategy, we develop a unique nanoparticle (NP) platform composed of a solid polymer/cationic lipid hybrid core and a lipid-poly(ethylene glycol) (lipid-PEG) shell for systemic siRNA delivery. The new generation lipid-polymer hybrid NPs are small and uniform, and can efficiently encapsulate siRNA and control its sustained release. They exhibit long blood circulation (t1/2 ∼ 8 h), high tumor accumulation, effective gene silencing, and negligible in vivo side effects. With this RNAi NP, we delineate and validate the therapeutic role of Prohibitin1 (PHB1), a target protein that has not been systemically evaluated in vivo due to the lack of specific and effective inhibitors, in treating non-small cell lung cancer (NSCLC) as evidenced by the drastic inhibition of tumor growth upon PHB1 silencing. Human tissue microarray analysis also reveals that high PHB1 tumor expression is associated with poorer overall survival in patients with NSCLC, further suggesting PHB1 as a therapeutic target. We expect this long-circulating RNAi NP platform to be of high interest for validating potential cancer targets in vivo and for the development of new cancer therapies.

  1. Long-circulating siRNA nanoparticles for validating Prohibitin1-targeted non-small cell lung cancer treatment

    PubMed Central

    Zhu, Xi; Xu, Yingjie; Solis, Luisa M.; Tao, Wei; Wang, Liangzhe; Behrens, Carmen; Xu, Xiaoyang; Zhao, Lili; Liu, Danny; Wu, Jun; Zhang, Ning; Wistuba, Ignacio I.; Farokhzad, Omid C.; Zetter, Bruce R.; Shi, Jinjun

    2015-01-01

    RNA interference (RNAi) represents a promising strategy for identification and validation of putative therapeutic targets and for treatment of a myriad of important human diseases including cancer. However, the effective systemic in vivo delivery of small interfering RNA (siRNA) to tumors remains a formidable challenge. Using a robust self-assembly strategy, we develop a unique nanoparticle (NP) platform composed of a solid polymer/cationic lipid hybrid core and a lipid-poly(ethylene glycol) (lipid-PEG) shell for systemic siRNA delivery. The new generation lipid–polymer hybrid NPs are small and uniform, and can efficiently encapsulate siRNA and control its sustained release. They exhibit long blood circulation (t1/2 ∼8 h), high tumor accumulation, effective gene silencing, and negligible in vivo side effects. With this RNAi NP, we delineate and validate the therapeutic role of Prohibitin1 (PHB1), a target protein that has not been systemically evaluated in vivo due to the lack of specific and effective inhibitors, in treating non-small cell lung cancer (NSCLC) as evidenced by the drastic inhibition of tumor growth upon PHB1 silencing. Human tissue microarray analysis also reveals that high PHB1 tumor expression is associated with poorer overall survival in patients with NSCLC, further suggesting PHB1 as a therapeutic target. We expect this long-circulating RNAi NP platform to be of high interest for validating potential cancer targets in vivo and for the development of new cancer therapies. PMID:26056316

  2. In vivo biodistribution of siRNA and cisplatin administered using CD44-targeted hyaluronic acid nanoparticles.

    PubMed

    Ganesh, Shanthi; Iyer, Arun K; Gattacceca, Florence; Morrissey, David V; Amiji, Mansoor M

    2013-12-28

    Multidrug resistance (MDR) is a significant problem in the clinical management of several cancers. Overcoming MDR generally involves multi-modal therapeutic approaches that integrate enhancement of delivery efficiency using targeted nano-platforms as well as strategies that can sensitize cancer cells to drug treatments. We recently demonstrated that tandem delivery of siRNAs that downregulate anti-apoptotic genes overexpressed in cisplatin resistant tumors followed by therapeutic challenge using cisplatin loaded CD44 targeted hyaluronic acid (HA) nanoparticle (NP) induced synergistic antitumor response CD44 expressing tumors that are resistant to cisplatin. In the current study, a near infrared (NIR) dye-loaded HA NP was employed to image the whole body localization of NPs after intravenous (i.v.) injection into live mice bearing human lung tumors that were sensitive and resistant to cisplatin. In addition, we quantified the siRNA duplexes and cisplatin dose distribution in various tissues and organs using an ultra-sensitive quantitative PCR method and inductively coupled plasma-mass spectrometry (ICP-MS), respectively, after i.v. injection of the payload loaded HA NPs in tumor bearing mice. Our findings demonstrate that the distribution pattern of the siRNA and cisplatin using specifically engineered CD44 targeting HA NPs correlated well with the tumor targeting capability as well as the activity and efficacy obtained with combination treatments.

  3. Efficient siRNA-peptide conjugation for specific targeted delivery into tumor cells.

    PubMed

    Gandioso, Albert; Massaguer, Anna; Villegas, Núria; Salvans, Cándida; Sánchez, Dani; Brun-Heath, Isabelle; Marchán, Vicente; Orozco, Modesto; Terrazas, Montserrat

    2017-03-02

    Despite the broad applicability of the Huisgen cycloaddition reaction, the click functionalization of RNAs with peptides still remains a challenge. Here we describe a straightforward method for the click functionalization of siRNAs with peptides of different sizes and complexities. Among them, a promising peptide carrier for the selective siRNA delivery into HER2+ breast cancer cell lines has been reported.

  4. Nanoengineered strategies for siRNA delivery: from target assessment to cancer therapeutic efficacy.

    PubMed

    Mishra, Dinesh Kumar; Balekar, Neelam; Mishra, Pradyumna Kumar

    2017-04-01

    The promise of RNA interference (RNAi) technology in cancer therapeutics aims to deliver small interfering RNA (siRNA) for silencing of gene expression in cell type-specific pathway. However, the challenge for the delivery of stable siRNA is hindered by an immune-hostile tumor microenvironment and physiological barriers of the circulatory system. Therefore, the development and validation of safe, stable, and efficient nanoengineered delivery systems are highly essential for effective delivery of siRNA into cancer cells. This review focuses on gene-silencing mechanisms, challenges to siRNA delivery, design and delivery of nanocarrier systems, ongoing clinical trials, and translational prospects for siRNA-mediated cancer therapeutics.

  5. Co-targeting EGFR and survivin with a bivalent aptamer-dual siRNA chimera effectively suppresses prostate cancer

    PubMed Central

    Liu, Hong Yan; Yu, Xiaolin; Liu, Haitao; Wu, Daqing; She, Jin-Xiong

    2016-01-01

    Current targeted therapies using small kinase inhibitors and antibodies have limited efficacy in treating prostate cancer (PCa), a leading cause of cancer death in American men. We have developed a novel strategy by engineering an RNA-based aptamer-siRNA chimera, in which a bivalent aptamer specifically binds prostate-specific membrane antigen (PSMA) via an antibody-like structure to promote siRNA internalization in PCa cells, and two siRNAs specific to EGFR and survivin are fused between two aptamers. The chimera is able to inhibit EGFR and survivin simultaneously and induce apoptosis effectively in vitro and in vivo. In the C4-2 PCa xenograft model, the treatment with the chimera significantly suppresses tumor growth and angiogenesis. The inhibition of angiogenesis is mediated by an EGFR-HIF1α-VEGF-dependent mechanism. Our results support that the bivalent aptamer-driven delivery of two siRNAs could be a new combination therapeutic strategy to effectively inhibit multiple and conventionally “undruggable” targets. PMID:27456457

  6. New therapeutic strategies in neuroblastoma: combined targeting of a novel tyrosine kinase inhibitor and liposomal siRNAs against ALK

    PubMed Central

    Di Paolo, Daniela; Yang, D.; Pastorino, Fabio; Emionite, Laura; Cilli, Michele; Daga, Antonio; Destefanis, Elisa; Di Fiore, Annarita; Piaggio, Francesca; Brignole, Chiara; Xu, Xiaobao; Liang, Chris; Gibbons, James

    2015-01-01

    Many different aberrations in the Anaplastic Lymphoma Kinase (ALK) were found to be oncogenic drivers in several cancers including neuroblastoma (NB), therefore ALK is now considered a critical player in NB oncogenesis and a promising therapeutic target. The ALK-inhibitor crizotinib has a limited activity against the various ALK mutations identified in NB patients. We tested: the activity of the novel ALK-inhibitor X-396 administered alone or in combination with Targeted Liposomes carrying ALK-siRNAs (TL[ALK-siRNA]) that are active irrespective of ALK gene mutational status; the pharmacokinetic profiles and the biodistribution of X-396; the efficacy of X-396 versus crizotinib treatment in NB xenografts; whether the combination of X-396 with the TL[ALK-siRNA] could promote long-term survival in NB mouse models. X-396 revealed good bioavailability, moderate half-life, high mean plasma and tumor concentrations. X-396 was more effective than crizotinib in inhibiting in vitro cell proliferation of NB cells and in reducing tumor volume in subcutaneous NB models in a dose-dependent manner. In orthotopic NB xenografts, X-396 significantly increased life span independently of the ALK mutation status. In combination studies, all effects were significantly improved in the mice treated with TL[ALK-siRNA] and X-396 compared to mice receiving the single agents. Our findings provide a rational basis to design innovative molecular-based treatment combinations for clinical application in ALK-driven NB tumors. PMID:26299615

  7. New therapeutic strategies in neuroblastoma: combined targeting of a novel tyrosine kinase inhibitor and liposomal siRNAs against ALK.

    PubMed

    Di Paolo, Daniela; Yang, D; Pastorino, Fabio; Emionite, Laura; Cilli, Michele; Daga, Antonio; Destafanis, Elisa; Di Fiore, Annarita; Piaggio, Francesca; Brignole, Chiara; Xu, Xiaobao; Liang, Chris; Gibbons, James; Ponzoni, Mirco; Perri, Patrizia

    2015-10-06

    Many different aberrations in the Anaplastic Lymphoma Kinase (ALK) were found to be oncogenic drivers in several cancers including neuroblastoma (NB), therefore ALK is now considered a critical player in NB oncogenesis and a promising therapeutic target. The ALK-inhibitor crizotinib has a limited activity against the various ALK mutations identified in NB patients. We tested: the activity of the novel ALK-inhibitor X-396 administered alone or in combination with Targeted Liposomes carrying ALK-siRNAs (TL[ALK-siRNA]) that are active irrespective of ALK gene mutational status; the pharmacokinetic profiles and the biodistribution of X-396; the efficacy of X-396 versus crizotinib treatment in NB xenografts; whether the combination of X-396 with the TL[ALK-siRNA] could promote long-term survival in NB mouse models. X-396 revealed good bioavailability, moderate half-life, high mean plasma and tumor concentrations. X-396 was more effective than crizotinib in inhibiting in vitro cell proliferation of NB cells and in reducing tumor volume in subcutaneous NB models in a dose-dependent manner. In orthotopic NB xenografts, X-396 significantly increased life span independently of the ALK mutation status. In combination studies, all effects were significantly improved in the mice treated with TL[ALK-siRNA] and X-396 compared to mice receiving the single agents. Our findings provide a rational basis to design innovative molecular-based treatment combinations for clinical application in ALK-driven NB tumors.

  8. siRNA delivery targeting to the lung via agglutination-induced accumulation and clearance of cationic tetraamino fullerene

    NASA Astrophysics Data System (ADS)

    Minami, Kosuke; Okamoto, Koji; Doi, Kent; Harano, Koji; Noiri, Eisei; Nakamura, Eiichi

    2014-05-01

    The efficient treatment of lung diseases requires lung-selective delivery of agents to the lung. However, lung-selective delivery is difficult because the accumulation of micrometer-sized carriers in the lung often induces inflammation and embolization-related toxicity. Here we demonstrate a lung-selective delivery system of small interfering RNA (siRNA) by controlling the size of carrier vehicle in blood vessels. The carrier is made of tetra(piperazino)fullerene epoxide (TPFE), a water-soluble cationic tetraamino fullerene. TPFE and siRNA form sub-micrometer-sized complexes in buffered solution and these complexes agglutinate further with plasma proteins in the bloodstream to form micrometer-sized particles. The agglutinate rapidly clogs the lung capillaries, releases the siRNA into lung cells to silence expression of target genes, and is then cleared rapidly from the lung after siRNA delivery. We applied our delivery system to an animal model of sepsis, indicating the potential of TPFE-based siRNA delivery for clinical applications.

  9. Targeted Delivery of siRNA with pH-Responsive Hybrid Gold Nanostars for Cancer Treatment.

    PubMed

    Zhu, Hongyan; Liu, Wanwan; Cheng, Ziting; Yao, Ke; Yang, Yu; Xu, Bohui; Su, Gaoxing

    2017-09-22

    In this work, we report the engineering of gold nanostars (GNS) to deliver small interfering RNA (siRNA) into HepG2 cells. The ligand DG-PEG-Lipoic acid (LA)-Lys-9R (hydrazone) was designed to functionalize GNS, and create the nanoparticles named as 9R/DG-GNS (hydrazone). In the ligand, 2-deoxyglucose (DG) is the targeting molecule, polyethylene glycol (PEG) helps to improve the dispersity and biocompatibility, 9-poly-d-arginine (9R) is employed to provide a positive surface charge and adsorb negative siRNA, and hydrazone bonds are pH-responsive and can avoid receptor-mediated endosomal recycling. Compared to GNS alone, 9R/DG-GNS (hydrazone) showed superior transfection efficiency. The expressions of cyclooxygenase-2 (COX-2) in HepG2 and SGC7901 cells were significantly suppressed by siRNA/9R/DG-GNS (hydrazone) complex. Notably, 9R/DG-GNS (hydrazone) possessed low cytotoxicity even at high concentrations in both normal cells and tumor cells. The combination treatment of siRNA/9R/DG-GNS (hydrazone) complex inhibited the cell growth rate by more than 75%. These results verified that the pH-responsive GNS complex is a promising siRNA delivery system for cancer therapy, and it is anticipated that near-infrared absorbing GNS with good photothermal conversion efficiency can be potentially used for photothermal therapy of tumors.

  10. Straining of SiGe ultrathin films with mesoporous Si substrates

    SciTech Connect

    Boucherif, A.; Guillot, G.; Lysenko, V.; Blanchard, N. P.; Regreny, P.; Grenet, G.; Marty, O.

    2010-09-27

    We report on the fabrication and characterization of ultrathin (down to 50 nm) tensile strained SiGe films on mesoporous Si substrates. Low temperature oxidation of the porous substrate relaxes the compressive strain in the as grown monocrystalline (mc) SiGe. Applying this method to a 50 nm thick mc-Si{sub 0.72}Ge{sub 0.28} film, a tensile strain >0.78% can be achieved without compromising crystalline quality and up to 1.45 % without the appearance of cracks.

  11. Fabrication of Source/Drain Electrodes for a-Si:H Thin-Film Transistors Using a Single Cu Alloy Target

    NASA Astrophysics Data System (ADS)

    Lee, J. H.; Lee, C. Y.; Nam, H. S.; Lee, J. G.; Yang, H. J.; Ho, W. J.; Jeong, J. Y.; Koo, D. H.

    2011-11-01

    A Cu alloy/Cu alloy oxide bilayer structure was formed on an n +-a-Si:H substrate using a single Cu alloy target. It was employed for the source/drain electrodes in the fabrication of a-Si:H thin-film transistors with good electrical performance, high thermal stability, and good adhesion. Transmission electron microscopy and electron energy-loss spectroscopy analyses revealed that the initial sputtering of the Cu alloy in O2/Ar allowed for preferential oxidation of Si and the formation of a SiO x /Cu-supersaturated a-Si:H bilayer at the copper oxide-a-Si:H interface. This bilayer turned into an SiO x /Cu3Si bilayer after annealing at 300°C. It provided a stable contact structure with low contact resistance.

  12. Multifunctional peptide-lipid nanocomplexes for efficient targeted delivery of DNA and siRNA into breast cancer cells.

    PubMed

    Wan, Yu; Dai, Wei; Nevagi, Reshma J; Toth, Istvan; Moyle, Peter M

    2017-09-01

    The development of carriers for the delivery of oligonucleotide therapeutics is essential for the successful translation of gene therapies to the clinic. In the present study, a delivery system, which combines the fusogenic lipid 1,2-dioleoyl-sn-glycero-3-phosphoethanolamine (DOPE) with a well-defined synthetic multifunctional peptide, was produced and optimized for gene delivery, with the aim to develop an efficient gene delivery platform for breast cancer cells. For this purpose, a breast cancer-specific cell targeting peptide (CTP) was incorporated into our leading peptide-based gene delivery system (to generate DEN-K(GALA)-TAT-K(STR)-CTP) to improve its cell-specific internalization, and investigated in combination with a formulation approach (DOPE/1,2-dioleoyl-3-trimethylammonium-propane (DOTAP)). DEN-K(GALA)-TAT-K(STR)-CTP alone efficiently complexed with DNA or siRNA, and promoted efficient cellular uptake, but low levels of gene expression. By adding the formulation approach, synergistic improvements in gene expression and silencing were observed compared to the peptide or formulation approaches alone. Of significance, this current system demonstrated more efficient gene knockdown when compared to the leading commercial siRNA delivery agent Lipofectamine® RNAiMAX. The utility of this system was demonstrated through the delivery of BCL2 (B-cell lymphoma 2) siRNA to MCF-7 cells, which led to near complete knockdown of the Bcl-2 protein, and inhibition of MCF-7 cell migration in a wound healing assay. The present peptide/lipid hybrid system is an excellent candidate for the delivery of DNA or siRNA into breast cancer cells. The identification of safe and effective delivery systems for DNA and siRNA is of great importance. Herein, we developed a well-defined, multifunctional and cell-specific lipidic peptide DEN-K(GALA)-TAT-K(STR)-CTP as a breast cancer cell targeted gene delivery vector. When combined with a lipid component (DOTAP/DOPE), the peptide

  13. Efficient Receptor Mediated siRNA Delivery in Vitro by Folic Acid Targeted Pentablock Copolymer-Based Micelleplexes.

    PubMed

    Lehner, Roman; Liu, Kegang; Wang, Xueya; Hunziker, Patrick

    2017-08-14

    Novel, biocompatible polyplexes, based on the combination of cationic pentablock copolymers with folic acid functionalized copolymers, were designed and developed for target-specific siRNA delivery. The resulting micelleplexes spontaneously formed polymeric micelles with a hydrophobic core surrounded directly by a cationic poly-2-(4-aminobutyl)-oxazole (PABOXA) and subsequently shielded by hydrophilic poly-2-methyl-oxazole (PMOXA) layer. The described micelleplexes form highly stable particles even in complete serum after 24 h compared with the highly cationic polymer PEI, which show aggregate formation in serum containing buffer solution. Targeted siRNA delivery and gene knockdown could be shown using green fluorescent protein (GFP) expressing HeLa cells, resulting in ∼31% and ∼8% suppression of the expression of GFP for targeted and nontargeted micelleplexes, respectively. Comparison studies of folic-receptor positive HeLa cells with normal folic-receptor-negative HEK293 cells revealed involvement of receptor mediated cellular uptake of fluorescently labeled siRNA. The new designed nanocarrier showed no cytotoxicity, having a potential application. The presented concept of shielding a nucleic-acid complexing cationic chains with a stealth layer and combining it with receptor ligand overcomes typical problems with undesired protein and cell interactions in delivery of nucleic acids using polymeric systems, opening new doors for application if RNA inhibition in the organism.

  14. Focused ultrasound for targeted delivery of siRNA and efficient knockdown of Htt expression.

    PubMed

    Burgess, Alison; Huang, Yuexi; Querbes, William; Sah, Dinah W; Hynynen, Kullervo

    2012-10-28

    RNA interference is a promising strategy for the treatment of Huntington's disease (HD) as it can specifically decrease the expression of the mutant Huntingtin protein (Htt). However, siRNA does not cross the blood-brain barrier and therefore delivery to the brain is limited to direct CNS delivery. Non-invasive delivery of siRNA through the blood-brain barrier (BBB) would be a significant advantage for translating this therapy to HD patients. Focused ultrasound (FUS), combined with intravascular delivery of microbubble contrast agent, was used to locally and transiently disrupt the BBB in the right striatum of adult rats. 48h following treatment with siRNA, the right (treated) and the left (control) striatum were dissected and analyzed for Htt mRNA levels. We demonstrate that FUS can non-invasively deliver siRNA-Htt directly to the striatum leading to a significant reduction of Htt expression in a dose dependent manner. Furthermore, we show that reduction of Htt with siRNA-Htt was greater when the extent of BBB disruption was increased. This study demonstrates that siRNA treatment for knockdown of mutant Htt is feasible without the surgical intervention previously required for direct delivery to the brain.

  15. Anisotropic etching of monocrystalline silicon under subcritical conditions

    NASA Astrophysics Data System (ADS)

    Gonzalez-Pereyra, Nestor Gabriel

    Sub- and supercritical fluids remain an underexploited resource for materials processing. Around its critical point a common compound such as water behaves like a different substance exhibiting changes in its properties that modify its behavior as a solvent and unlock reaction paths not viable in other conditions. In the subcritical region water's properties can be directed by controlling temperature and pressure. Water and silicon are two of the most abundant, versatile, environmentally non-harmful, and simplest substances on Earth. They are among the most researched and best-known substances. Both are ubiquitous and essential for present-day world. Silicon is fundamental in semiconductor fabrication, microelectromechanical systems, and photovoltaic cells. Wet etching of silicon is a fabrication strategy shared by these three applications. Processing of silicon requires large amounts of water, often involving dangerous and environmentally hazardous chemicals. Yet, minimal knowledge is available on the ways high temperature water interacts with crystalline silicon. The purpose of this project is to identify and implement a method for the modification of monocrystalline silicon surfaces with three important characteristics: 1) requires minimal amounts of added chemicals, 2) controllability of morphological features formed, 3) reduced processing time. This will be accomplished by subjecting crystalline silicon to diluted alkaline solutions working in the subcritical region of water. This approach allows for variations on surface morphologies and etching rates by adapting the reactions conditions, with focus on composition and temperature of the solutions used. The work reported discusses the techniques used for producing surfaces with a variety of morphologies that ultimately allowed to create patterns and textures on silicon wafers, using highly diluted alkaline solutions that can be used for photovoltaic applications. These morphologies were created with a

  16. Triple combination of siRNAs targeting TGFβ1, TGFβR2, and CTGF enhances reduction of collagen I and smooth muscle actin in corneal fibroblasts.

    PubMed

    Sriram, Sriniwas; Robinson, Paulette; Pi, Liya; Lewin, Alfred S; Schultz, Gregory

    2013-12-17

    Transforming growth factor β1 (TGFβ1), TGFβ receptor (TGFβR2), and connective tissue growth factor (CTGF) are key regulators of fibrosis in the cornea and in other tissues, including liver, skin, and kidney. We developed an antifibrotic treatment targeting these three critical scarring genes by using a combination of small interfering RNAs (siRNAs) and assessed its effect on downstream scarring genes, collagen I, and α smooth muscle actin (SMA). Up to six individual siRNAs for each of the three target gene mRNAs were transfected into cultures of rabbit corneal fibroblasts at concentrations from 15 to 90 nM. The knockdown of target gene proteins was measured by ELISA, and the two most effective siRNAs were tested in dual combinations. Knockdown percentages of both individual and dual siRNA combinations were analyzed for synergy by using combination index to predict "effective" and "ineffective" triple siRNA combinations. Effects of both triple siRNA combinations on target and downstream mRNAs were measured by using quantitative RT-PCR, and levels of SMA protein were assessed by immunohistochemistry. Single and dual siRNA combinations produced a wide range of protein knockdown of target genes (5%-80%). The effective triple siRNA combination significantly reduced mRNA levels of target genes (>80%) and downstream scarring genes (>85%), and of SMA protein (>95%), and significantly reduced cell migration without reducing cell viability. Simultaneous targeting of TGFβ1, TGFβR2, and CTGF genes by effective triple siRNA combination produced high knockdown of target and downstream scarring genes without cell toxicity, which may have clinical applications in reducing corneal fibrosis and scarring in other tissues.

  17. Triple Combination of siRNAs Targeting TGFβ1, TGFβR2, and CTGF Enhances Reduction of Collagen I and Smooth Muscle Actin in Corneal Fibroblasts

    PubMed Central

    Sriram, Sriniwas; Robinson, Paulette; Pi, Liya; Lewin, Alfred S.; Schultz, Gregory

    2013-01-01

    Purpose. Transforming growth factor β1 (TGFβ1), TGFβ receptor (TGFβR2), and connective tissue growth factor (CTGF) are key regulators of fibrosis in the cornea and in other tissues, including liver, skin, and kidney. We developed an antifibrotic treatment targeting these three critical scarring genes by using a combination of small interfering RNAs (siRNAs) and assessed its effect on downstream scarring genes, collagen I, and α smooth muscle actin (SMA). Methods. Up to six individual siRNAs for each of the three target gene mRNAs were transfected into cultures of rabbit corneal fibroblasts at concentrations from 15 to 90 nM. The knockdown of target gene proteins was measured by ELISA, and the two most effective siRNAs were tested in dual combinations. Knockdown percentages of both individual and dual siRNA combinations were analyzed for synergy by using combination index to predict “effective” and “ineffective” triple siRNA combinations. Effects of both triple siRNA combinations on target and downstream mRNAs were measured by using quantitative RT-PCR, and levels of SMA protein were assessed by immunohistochemistry. Results. Single and dual siRNA combinations produced a wide range of protein knockdown of target genes (5%–80%). The effective triple siRNA combination significantly reduced mRNA levels of target genes (>80%) and downstream scarring genes (>85%), and of SMA protein (>95%), and significantly reduced cell migration without reducing cell viability. Conclusions. Simultaneous targeting of TGFβ1, TGFβR2, and CTGF genes by effective triple siRNA combination produced high knockdown of target and downstream scarring genes without cell toxicity, which may have clinical applications in reducing corneal fibrosis and scarring in other tissues. PMID:24282226

  18. Roles of target site location and sequence complementarity in trans-acting siRNA formation in Arabidopsis.

    PubMed

    Zhang, Changqing; Ng, Danny W-K; Lu, Jie; Chen, Z Jeffrey

    2012-01-01

    In plants, many mRNAs and non-coding RNAs are cleaved by RNA-induced silencing complexes. After cleavage, only a limited number of RNAs are processed into trans-acting siRNAs (tasiRNAs). One reason is that 22 nt small RNAs, but not the more common 21 nt small RNAs, can efficiently trigger tasiRNA formation. The characteristics of the target transcripts may also affect tasiRNA production. Here we report the effects of target site location and sequence complementarity on tasiRNA formation. A synthetic sequence that included a miR173 target site and two siRNAs targeting an endogenous mRNA encoding PHYTOENE DESATURASE3 was introduced into a protein-coding (GFP) gene in the coding region or 3' UTR. tasiRNAs were generated in the transgenic seedlings, and the PDS3 mRNA level was reduced, leading to a photobleaching phenotype. It was found that tasiRNAs were most efficiently produced when the miR173 target site was placed immediately after the stop codon. Introducing premature stop codons caused a dramatic reduction of tasiRNAs and over-accumulation of 3' cleavage products, suggesting positive effects of translation on processing the 3' cleavage products into tasiRNAs. By systematically mutating the miR173 target site, we found that perfect complementarity between the 3' end of miR173 and the 5' end of the target sequence was crucial. Mismatches at that position abolished tasiRNA formation, but mismatches at the 5' end of miR173 had less effect. These data suggest important roles for translation and specific sequence complementarity in tasiRNA formation, providing new insights into tasiRNA biogenesis as well as a strategy for improving the efficiency of RNA interference (RNAi) using tasiRNAs. © 2011 The Authors. The Plant Journal © 2011 Blackwell Publishing Ltd.

  19. Lipoprotein-biomimetic nanostructure enables efficient targeting delivery of siRNA to Ras-activated glioblastoma cells via macropinocytosis

    NASA Astrophysics Data System (ADS)

    Huang, Jia-Lin; Jiang, Gan; Song, Qing-Xiang; Gu, Xiao; Hu, Meng; Wang, Xiao-Lin; Song, Hua-Hua; Chen, Le-Pei; Lin, Ying-Ying; Jiang, Di; Chen, Jun; Feng, Jun-Feng; Qiu, Yong-Ming; Jiang, Ji-Yao; Jiang, Xin-Guo; Chen, Hong-Zhuan; Gao, Xiao-Ling

    2017-05-01

    Hyperactivated Ras regulates many oncogenic pathways in several malignant human cancers including glioblastoma and it is an attractive target for cancer therapies. Ras activation in cancer cells drives protein internalization via macropinocytosis as a key nutrient-gaining process. By utilizing this unique endocytosis pathway, here we create a biologically inspired nanostructure that can induce cancer cells to `drink drugs' for targeting activating transcription factor-5 (ATF5), an overexpressed anti-apoptotic transcription factor in glioblastoma. Apolipoprotein E3-reconstituted high-density lipoprotein is used to encapsulate the siRNA-loaded calcium phosphate core and facilitate it to penetrate the blood-brain barrier, thus targeting the glioblastoma cells in a macropinocytosis-dependent manner. The nanostructure carrying ATF5 siRNA exerts remarkable RNA-interfering efficiency, increases glioblastoma cell apoptosis and inhibits tumour cell growth both in vitro and in xenograft tumour models. This strategy of targeting the macropinocytosis caused by Ras activation provides a nanoparticle-based approach for precision therapy in glioblastoma and other Ras-activated cancers.

  20. Lipoprotein-biomimetic nanostructure enables efficient targeting delivery of siRNA to Ras-activated glioblastoma cells via macropinocytosis

    PubMed Central

    Huang, Jia-Lin; Jiang, Gan; Song, Qing-Xiang; Gu, Xiao; Hu, Meng; Wang, Xiao-Lin; Song, Hua-Hua; Chen, Le-Pei; Lin, Ying-Ying; Jiang, Di; Chen, Jun; Feng, Jun-Feng; Qiu, Yong-Ming; Jiang, Ji-Yao; Jiang, Xin-Guo; Chen, Hong-Zhuan; Gao, Xiao-Ling

    2017-01-01

    Hyperactivated Ras regulates many oncogenic pathways in several malignant human cancers including glioblastoma and it is an attractive target for cancer therapies. Ras activation in cancer cells drives protein internalization via macropinocytosis as a key nutrient-gaining process. By utilizing this unique endocytosis pathway, here we create a biologically inspired nanostructure that can induce cancer cells to ‘drink drugs' for targeting activating transcription factor-5 (ATF5), an overexpressed anti-apoptotic transcription factor in glioblastoma. Apolipoprotein E3-reconstituted high-density lipoprotein is used to encapsulate the siRNA-loaded calcium phosphate core and facilitate it to penetrate the blood–brain barrier, thus targeting the glioblastoma cells in a macropinocytosis-dependent manner. The nanostructure carrying ATF5 siRNA exerts remarkable RNA-interfering efficiency, increases glioblastoma cell apoptosis and inhibits tumour cell growth both in vitro and in xenograft tumour models. This strategy of targeting the macropinocytosis caused by Ras activation provides a nanoparticle-based approach for precision therapy in glioblastoma and other Ras-activated cancers. PMID:28489075

  1. Microstructure and temperature coefficient of resistance of thin cermet resistor films deposited from CrSi{sub 2}-Cr-SiC targets by S-gun magnetron

    SciTech Connect

    Felmetsger, Valery V.

    2010-01-15

    Technological solutions for producing nanoscale cermet resistor films with sheet resistances above 1000 {Omega}/{open_square} and low temperature coefficients of resistance (TCR) have been investigated. 2-40 nm thick cermet films were sputter deposited from CrSi{sub 2}-Cr-SiC targets by a dual cathode dc S-gun magnetron. In addition to studying film resistance versus temperature, the nanofilm structural features and composition were analyzed using scanning electron microscopy, atomic force microscopy, high-resolution transmission electron microscopy, energy-dispersive x-ray spectroscopy, and electron energy loss spectroscopy. This study has revealed that all cermet resistor films deposited at ambient and elevated temperatures were amorphous. The atomic ratio of Si to Cr in these films was about 2 to 1. The film TCR displayed a significant increase when the deposited film thickness was reduced below 2.5 nm. An optimized sputter process consisting of wafer degassing, cermet film deposition at elevated temperature with rf substrate bias, and a double annealing in vacuum, consisting of in situ annealing following the film sputtering and an additional annealing following the exposure of the wafers to air, has been found to be very effective for the film thermal stabilization and for fine tuning the film TCR. Cermet films with thicknesses in the range of 2.5-4 nm deposited using this technique had sheet resistances ranging from 1800 to 1200 {Omega}/{open_square} and TCR values from -50 ppm/ deg. C to near zero, respectively. A possible mechanism responsible for the high efficiency of annealing the cermet films in vacuum (after preliminary exposure to air), resulting in resistance stabilization and TCR reduction, is also discussed.

  2. Targeted siRNA delivery by anti-HER2 antibody-modified nanoparticles of mPEG-chitosan diblock copolymer.

    PubMed

    Wang, Yiyi; Liu, Peifeng; Du, Jing; Sun, Ying; Li, Fenghua; Duan, Yourong

    2013-01-01

    This study aims to determine the specificity of anti-human epidermal growth factor receptor antibody (anti-HER2) modified monomethoxy polyethylene glycol-chitosan (mPEG-CS) nanoparticles (anti-HER2/mPEG-CS NPs) in delivering small interfering RNA (siRNA) to the human epidermal growth factor receptor 2 (HER2) positive cancer cells. Physicochemical properties of the siRNA-loaded anti-HER2/mPEG-CS NPs (anti-HER2/mPEG-CS-siRNA NPs), including size, surface charge, siRNA encapsulation efficiency, and in vitro release profile of siRNA from NPs, were characterized by particle size and zeta potential analyzer, and ultraviolet-visible spectrophotometer. MTT assay was used to study the in vitro cytotoxicity of the NPs. Fluorescent microscope and flow cytometer analysis results showed that anti-HER2/mPEG-CS-siRNA NPs had much efficient delivery of siRNA than the siRNA alone, Lipofectamine-siRNA complexes and mPEG-CS-siRNA NPs. These results demonstrated that anti-HER2/mPEG-CS-siRNA NPs had great potential applications as a targeted strategy for siRNA delivery.

  3. Fabrication of Fe@mSiO2 nanowires with large remanence and low cytotoxicity for targeted drug delivery

    NASA Astrophysics Data System (ADS)

    Song, Meng-Meng; Bi, Hong; Zhang, Ye

    2012-04-01

    Core-shell structured Fe@mSiO2 nanowires for targeted drug delivery have been prepared through electrodeposition followed by a CTAB-template sol-gel process. The magnetic Fe nanowire core has a diameter of ˜40 nm and the mesoporous silica shell has a uniform thickness of ˜40 nm with an average pore size of ˜2.45 nm. The drug loading experiment indicates Fe@mSiO2 nanowires have a good capability for loading drug molecules due to the large surface area of the mesoporous silica shell. Furthermore, MDA-MB-231 human breast cancer cells were chosen as model cells to investigate cyototoxicity of the nanowires by 3-(4,5)-dimethylthiahiazo (-z-y1)-3,5-di-phenytetrazoliumromide (MTT) and lactate dehydrogenase (LDH) assays. MTT results show low cytotoxicity of the nanowires, which is concentration-dependent and the cell viability is still nearly 80% while the concentration reaches as high as 250 μg/mL. Moreover, LDH assay has demonstrated that the nanowires have no influence on the integrity of the cell membrane. All results indicate that the as-prepared Fe@mSiO2 nanowires have a potential application as a drug nanocarrier for magnetic-targeted drug delivery.

  4. Polysaccharide Nanoparticles for Efficient siRNA Targeting in Cancer Cells by Supramolecular pKa Shift

    PubMed Central

    Zhang, Ying-Ming; Yang, Yang; Zhang, Yu-Hui; Liu, Yu

    2016-01-01

    Biomacromolecular pKa shifting is considered as one of the most ubiquitous processes in biochemical events, e.g., the enzyme-catalyzed reaction and protein conformational stabilization. In this paper, we report on the construction of biocompatible polysaccharide nanoparticle with targeting ability and lower toxicity by supramolecular pKa shift strategy. This was realized through a ternary assembly constructed by the dual host‒guest interactions of an adamantane-bis(diamine) conjugate (ADA) with cucurbit[6]uril (CB[6]) and a polysaccharide. The potential application of such biocompatible nanostructure was further implemented by the selective transportation of small interfering RNA (siRNA) in a controlled manner. It is demonstrated that the strong encapsulation of the ADA’s diammonium tail by CB[6] not only reduced the cytotoxicity of the nano-scaled vehicle but also dramatically enhanced cation density through an obvious positive macrocycle-induced pKa shift, which eventually facilitated the subsequent siRNA binding. With a targeted polysaccharide shell containing a cyclodextrin‒hyaluronic acid conjugate, macrocycle-incorporated siRNA polyplexes were specifically delivered into malignant human prostate PC-3 cells. The supramolecular polysaccharide nanoparticles, the formation of which was enabled and promoted by the complexation-assisted pKa shift, may be used as a versatile tool for controlled capture and release of biofunctional substrates. PMID:27363811

  5. EGF receptor targeted lipo-oligocation polyplexes for antitumoral siRNA and miRNA delivery

    NASA Astrophysics Data System (ADS)

    Müller, Katharina; Klein, Philipp M.; Heissig, Philipp; Roidl, Andreas; Wagner, Ernst

    2016-11-01

    Antitumoral siRNA and miRNA delivery was demonstrated by epidermal growth factor receptor (EGFR) targeted oligoaminoamide polyplexes. For this purpose, the T-shaped lipo-oligomer 454 was used to complex RNA into a core polyplex, which was subsequently functionalized with the targeting peptide ligand GE11 via a polyethylene glycol (PEG) linker. To this end, free cysteines on the surface of 454 polyplex were coupled with a maleimide-PEG-GE11 reagent (Mal-GE11). Resulting particles with sizes of 120-150 nm showed receptor-mediated uptake into EGFR-positive T24 bladder cancer cells, MDA-MB 231 breast cancer cells and Huh7 liver cancer cells. Furthermore, these formulations led to ligand-dependent gene silencing. RNA interference (RNAi) triggered antitumoral effects were observed for two different therapeutic RNAs, a miRNA-200c mimic or EG5 siRNA. Using polyplexes modified with a ratio of 0.8 molar equivalents of Mal-GE11, treatment of T24 or MDA-MB 231 cancer cells with miR-200c led to the expected decreased proliferation and migration, changes in cell cycle and enhanced sensitivity towards doxorubicin. Delivery of EG5 siRNA into Huh7 cells resulted in antitumoral activity with G2/M arrest, triggered by loss of mitotic spindle separation and formation of mono-astral spindles. These findings demonstrate the potential of GE11 ligand-containing RNAi polyplexes for cancer treatment.

  6. Polysaccharide Nanoparticles for Efficient siRNA Targeting in Cancer Cells by Supramolecular pKa Shift

    NASA Astrophysics Data System (ADS)

    Zhang, Ying-Ming; Yang, Yang; Zhang, Yu-Hui; Liu, Yu

    2016-07-01

    Biomacromolecular pKa shifting is considered as one of the most ubiquitous processes in biochemical events, e.g., the enzyme-catalyzed reaction and protein conformational stabilization. In this paper, we report on the construction of biocompatible polysaccharide nanoparticle with targeting ability and lower toxicity by supramolecular pKa shift strategy. This was realized through a ternary assembly constructed by the dual host‒guest interactions of an adamantane-bis(diamine) conjugate (ADA) with cucurbit[6]uril (CB[6]) and a polysaccharide. The potential application of such biocompatible nanostructure was further implemented by the selective transportation of small interfering RNA (siRNA) in a controlled manner. It is demonstrated that the strong encapsulation of the ADA’s diammonium tail by CB[6] not only reduced the cytotoxicity of the nano-scaled vehicle but also dramatically enhanced cation density through an obvious positive macrocycle-induced pKa shift, which eventually facilitated the subsequent siRNA binding. With a targeted polysaccharide shell containing a cyclodextrin‒hyaluronic acid conjugate, macrocycle-incorporated siRNA polyplexes were specifically delivered into malignant human prostate PC-3 cells. The supramolecular polysaccharide nanoparticles, the formation of which was enabled and promoted by the complexation-assisted pKa shift, may be used as a versatile tool for controlled capture and release of biofunctional substrates.

  7. Disruption of Aedes aegypti Olfactory System Development through Chitosan/siRNA Nanoparticle Targeting of semaphorin-1a

    PubMed Central

    Mysore, Keshava; Flannery, Ellen M.; Tomchaney, Michael; Severson, David W.; Duman-Scheel, Molly

    2013-01-01

    Despite the devastating impact of mosquito-borne illnesses on human health, surprisingly little is known about mosquito developmental biology, including development of the olfactory system, a tissue of vector importance. Analysis of mosquito olfactory developmental genetics has been hindered by a lack of means to target specific genes during the development of this sensory system. In this investigation, chitosan/siRNA nanoparticles were used to target semaphorin-1a (sema1a) during olfactory system development in the dengue and yellow fever vector mosquito Aedes aegypti. Immunohistochemical analyses and anterograde tracing of antennal sensory neurons, which were used to track the progression of olfactory development in this species, revealed antennal lobe defects in sema1a knockdown fourth instar larvae. These findings, which correlated with a larval odorant tracking behavioral phenotype, identified previously unreported roles for Sema1a in the developing insect larval olfactory system. Analysis of sema1a knockdown pupae also revealed a number of olfactory phenotypes, including olfactory receptor neuron targeting and projection neuron defects coincident with a collapse in the structure and shape of the antennal lobe and individual glomeruli. This study, which is to our knowledge the first functional genetic analysis of insect olfactory development outside of D. melanogaster, identified critical roles for Sema1a during Ae. aegypti larval and pupal olfactory development and advocates the use of chitosan/siRNA nanoparticles as an effective means of targeting genes during post-embryonic Ae. aegypti development. Use of siRNA nanoparticle methodology to understand sensory developmental genetics in mosquitoes will provide insight into the evolutionary conservation and divergence of key developmental genes which could be exploited in the development of both common and species-specific means for intervention. PMID:23696908

  8. A new texturing technique of monocrystalline silicon surface with sodium hypochlorite

    NASA Astrophysics Data System (ADS)

    Sun, Linfeng; Tang, Jiuyao

    2009-08-01

    This work proposes a new texturing technique of monocrystalline silicon surface for solar cells with sodium hypochlorite. A mixed solution consisting of 5 wt% sodium hypochlorite and 10 vl% ethanol has been found that results in a homogeneous pyramidal structure, and an optimal size of pyramids on the silicon surface. The textured silicon surface exhibits a lower average reflectivity (about 10.8%) in the main range of solar spectrum (400-1000 nm).

  9. Modified gold-based siRNA nanotherapeutics for targeted therapy of triple-negative breast cancer.

    PubMed

    Shahbazi, Reza; Asik, Elif; Kahraman, Nermin; Turk, Mustafa; Ozpolat, Bulent; Ulubayram, Kezban

    2017-08-01

    In this study, we aimed to therapeutically target eukaryotic elongation factor 2 kinase (eEF-2K) in an in vivo triple-negative breast cancer (TNBC) tumor model. We synthesized a highly monodisperse nanoformulation using polyethylenimine-modified gold nanoparticles (AuNP-PEI) as siRNA delivery vehicle and evaluated gene downregulation. We found that AuNP-PEI/eEF-2K nanoformulation was highly effective for in vitro and in vivo gene downregulation and showed remarkable antitumor efficacy that was associated with eEF-2K knockdown, inhibition of Src and MAPK-ERK signaling pathways in a TNBC orthotopic tumor model. Our study suggests that eEF-2K plays an important role in TNBC tumorigenesis and its inhibition by AuNP-PEI/eEF-2K siRNA-based nanotherapeutics may be a potential therapeutic strategy for TNBC.

  10. Use of Monocrystalline Silicon as Tool Material for Highly Accurate Blanking of Thin Metal Foils

    SciTech Connect

    Hildering, Sven; Engel, Ulf; Merklein, Marion

    2011-05-04

    The trend towards miniaturisation of metallic mass production components combined with increased component functionality is still unbroken. Manufacturing these components by forming and blanking offers economical and ecological advantages combined with the needed accuracy. The complexity of producing tools with geometries below 50 {mu}m by conventional manufacturing methods becomes disproportional higher. Expensive serial finishing operations are required to achieve an adequate surface roughness combined with accurate geometry details. A novel approach for producing such tools is the use of advanced etching technologies for monocrystalline silicon that are well-established in the microsystems technology. High-precision vertical geometries with a width down to 5 {mu}m are possible. The present study shows a novel concept using this potential for the blanking of thin copper foils with monocrystallline silicon as a tool material. A self-contained machine-tool with compact outer dimensions was designed to avoid tensile stresses in the brittle silicon punch by an accurate, careful alignment of the punch, die and metal foil. A microscopic analysis of the monocrystalline silicon punch shows appropriate properties regarding flank angle, edge geometry and surface quality for the blanking process. Using a monocrystalline silicon punch with a width of 70 {mu}m blanking experiments on as-rolled copper foils with a thickness of 20 {mu}m demonstrate the general applicability of this material for micro production processes.

  11. Fabrication of high resolution and lightweight monocrystalline silicon x-ray mirrors

    NASA Astrophysics Data System (ADS)

    Riveros, Raul E.; Kolos, Linette D.; Mazzarella, James R.; McKeon, Kevin P.; Zhang, William W.

    2015-09-01

    Monocrystalline silicon as an x-ray mirror substrate material promises significant improvements over the x- ray mirror technologies used to date, since it is mechanically stiff, stress-free, highly thermally conductive, and widely commercially available. Producing highly accurate and lightweight x-ray mirrors from monocrystalline silicon requires a unique and specialized manufacturing process capable of producing mirrors quickly and cost effectively. The identification, development, and testing of this process is the focus of the work described in this proceeding. Monocrystalline silicon blocks were obtained, and a variety of processes (wire electro-discharge machining, etching, polishing) were applied to generate an accurate and stress-free cylindrical or Wolter-I mirror surface. The mirror surface is then sliced off at a thickness of <1 mm and further processed to yield a mirror segment with <1 arcsecond RMS slope errors. Furthermore, our experiments suggest that this mirror production process requires ~2 days to produce a mirror segment and is easily integrated into a cost-reducing parallel processing scheme. Presently, there is strong evidence that the mirror production process described in this paper will meet the stringent requirements of future x-ray missions.

  12. Growth, Characterization and Device Development in Monocrystalline Diamond Films

    DTIC Science & Technology

    1990-09-30

    silicon substrate surfaces. The aspect ratio of diamond grown on single crystals of different substrate materials correlates with their surface free ...to check the feasibility of such considerations for the present research. The surface free energy of diamond is estimated to be 3387 ergs/cm 2, [Il...while the surface free energies of Si, Ni, Mo, Ta, and W are 1457 ergs/cm2 , 2072 ergs/cm 2, 2463 ergs/cm 2 , 2628 ergs/cm 2 and 3111 ergs/cm 2

  13. Targeting Intestinal Inflammation With CD98 siRNA/PEI–loaded Nanoparticles

    PubMed Central

    Laroui, Hamed; Geem, Duke; Xiao, Bo; Viennois, Emilie; Rakhya, Poonam; Denning, Timothy; Merlin, Didier

    2014-01-01

    Intestinal CD98 expression plays a crucial role in controlling homeostatic and innate immune responses in the gut. Modulation of CD98 expression in intestinal cells therefore represents a promising therapeutic strategy for the treatment and prevention of inflammatory intestinal diseases, such as inflammatory bowel disease. Here, the advantages of nanoparticles (NPs) are used, including their ability to easily pass through physiological barriers and evade phagocytosis, high loading concentration, rapid kinetics of mixing and resistance to degradation. Using physical chemistry characterizations techniques, CD98 siRNA/polyethyleneimine (PEI)–loaded NPs was characterized (diameter of ~480 nm and a zeta potential of −5.26 mV). Interestingly, CD98 siRNA can be electrostatically complexed by PEI and thus protected from RNase. In addition, CD98 siRNA/PEI–loaded NPs are nontoxic and biocompatible with intestinal cells. Oral administration of CD98/PEI-loaded NPs encapsulated in a hydrogel reduced CD98 expression in mouse colonic tissues and decreased dextran sodium sulfate–induced colitis in a mouse model. Finally, flow cytometry showed that CD98 was effectively downregulated in the intestinal epithelial cells and intestinal macrophages of treated mice. Finally, the results collectively demonstrated the therapeutic effect of “hierarchical nano-micro particles” with colon-homing capabilities and the ability to directly release “molecularly specific” CD98 siRNA in colonic cells, thereby decreasing colitis. PMID:24025751

  14. Multifunctional polyglycerol-grafted Fe₃O₄@SiO₂ nanoparticles for targeting ovarian cancer cells.

    PubMed

    Wang, Liang; Neoh, Koon Gee; Kang, En-Tang; Shuter, Borys

    2011-03-01

    Ligand-mediated magnetic resonance (MR) contrast agents would be highly desirable for cancer diagnosis. In the present study, nanoparticles of Fe₃O₄ core with fluorescent SiO₂ shell were synthesized and grafted with hyperbranched polyglycerol (HPG-grafted Fe₃O₄@SiO₂ nanoparticles). These nanoparticles have a hydrodynamic diameter of 47.0 ± 4.0 nm, and are very stable in aqueous solution as well as in cell culture medium. Numerous surface hydroxyl groups of these nanoparticles were conjugated with folic acid by a thiol 'click' reaction. The successful covalent attachment of folic acid on the nanoparticles was confirmed by FTIR and XPS analyses. Both MR imaging and fluorescence microscopy show significant preferential uptake of the folic acid-conjugated polyglycerol-grafted Fe₃O₄@SiO₂ (FA-HPG-grafted Fe₃O₄@SiO₂) nanoparticles by human ovarian carcinoma cells (SKOV-3) as compared to macrophages and fibroblasts. Such nanoparticles can potentially be used to provide real-time imaging in ovarian cancer resection.

  15. A tumor-targeting cRGD-EGFR siRNA conjugate and its anti-tumor effect on glioblastoma in vitro and in vivo.

    PubMed

    He, Shuai; Cen, Bohong; Liao, Lumin; Wang, Zhen; Qin, Yixin; Wu, Zhuomin; Liao, Wenjie; Zhang, Zhongyi; Ji, Aimin

    2017-11-01

    The epidermal growth factor receptor (EGFR) is an important anti-tumor target. The development of novel molecular-targeted anti-tumor drugs that can target the interior of tumor cells and specifically silence EGFR expression is valuable and promising. In this work, a promising anti-tumor conjugate comprising methoxy-modified EGFR siRNA and cyclic arginine-glycine-aspartic acid (cRGD) peptides, which selectively bind to αvβ3 integrins, was synthesized and examined. To prepare cRGD-EGFR siRNA (cRGD-siEGFR), cRGD was covalently conjugated to the 5'-end of an siRNA sense strand using a thiol-maleimide linker. The cellular uptake and cytotoxicity of cRGD-siEGFR in vitro were tested using an αvβ3-positive U87MG cell line. In vivo bio-distribution, anti-tumor activity, immunogenicity and toxicity were investigated in a nude mouse tumor model through repeated i.v. administration of cRGD-siEGFR (7 times over a 48 h interval). Analyses of in vitro data showed that cRGD-siEGFR silenced EGFR expression effectively, with high tumor targeting ability. Administration of cRGD-siEGFR to tumor-bearing nude mice led to significant inhibition of tumor growth, obvious reduction of EGFR expression and down-regulation of EGFR mRNA and protein in tumor tissue. Furthermore, serum biochemistry and pathological section evaluation did not indicate any serious toxicity of cRGD-siEGFR in vivo. cRGD-siEGFR is likely a promising candidate with high targeting ability, substantial anti-tumor effects and low toxicity in vitro and in vivo.

  16. Impact of tumor-specific targeting on the biodistribution and efficacy of siRNA nanoparticles measured by multimodality in vivo imaging.

    PubMed

    Bartlett, Derek W; Su, Helen; Hildebrandt, Isabel J; Weber, Wolfgang A; Davis, Mark E

    2007-09-25

    Targeted delivery represents a promising approach for the development of safer and more effective therapeutics for oncology applications. Although macromolecules accumulate nonspecifically in tumors through the enhanced permeability and retention (EPR) effect, previous studies using nanoparticles to deliver chemotherapeutics or siRNA demonstrated that attachment of cell-specific targeting ligands to the surface of nanoparticles leads to enhanced potency relative to nontargeted formulations. Here, we use positron emission tomography (PET) and bioluminescent imaging to quantify the in vivo biodistribution and function of nanoparticles formed with cyclodextrin-containing polycations and siRNA. Conjugation of 1,4,7,10-tetraazacyclododecane-1,4,7,10-tetraacetic acid to the 5' end of the siRNA molecules allows labeling with (64)Cu for PET imaging. Bioluminescent imaging of mice bearing luciferase-expressing Neuro2A s.c. tumors before and after PET imaging enables correlation of functional efficacy with biodistribution data. Although both nontargeted and transferrin-targeted siRNA nanoparticles exhibit similar biodistribution and tumor localization by PET, transferrin-targeted siRNA nanoparticles reduce tumor luciferase activity by approximately 50% relative to nontargeted siRNA nanoparticles 1 d after injection. Compartmental modeling is used to show that the primary advantage of targeted nanoparticles is associated with processes involved in cellular uptake in tumor cells rather than overall tumor localization. Optimization of internalization may therefore be key for the development of effective nanoparticle-based targeted therapeutics.

  17. Polarized 3He target and Final State Interactions in SiDIS

    DOE PAGES

    Del Dotto, Alessio; Kaptari, Leonid; Pace, Emanuele; ...

    2017-01-03

    Jefferson Lab is starting a wide experimental program aimed at studying the neutron’s structure, with a great emphasis on the extraction of the parton transverse-momentum distributions (TMDs). To this end, Semi-inclusive deep-inelastic scattering (SiDIS) experiments on polarized $^3$He will be carried out, providing, together with proton and deuteron data, a sound flavor decomposition of the TMDs. Here, given the expected high statistical accuracy, it is crucial to disentangle nuclear and partonic degrees of freedom to get an accurate theoretical description of both initial and final states. In this contribution, a preliminary study of the Final State Interaction (FSI) in themore » standard SiDIS, where a pion (or a Kaon) is detected in the final state is presented, in view of constructing a realistic description of the nuclear initial and final states.« less

  18. Polarized ^{\\varvec{3}}He Target and Final State Interactions in SiDIS

    NASA Astrophysics Data System (ADS)

    Del Dotto, Alessio; Kaptari, Leonid; Pace, Emanuele; Salmè, Giovanni; Scopetta, Sergio

    2017-01-01

    Jefferson Lab is starting a wide experimental program aimed at studying the neutron's structure, with a great emphasis on the extraction of the parton transverse-momentum distributions (TMDs). To this end, Semi-inclusive deep-inelastic scattering (SiDIS) experiments on polarized ^3He will be carried out, providing, together with proton and deuteron data, a sound flavor decomposition of the TMDs. Given the expected high statistical accuracy, it is crucial to disentangle nuclear and partonic degrees of freedom to get an accurate theoretical description of both initial and final states. In this contribution, a preliminary study of the Final State Interaction (FSI) in the standard SiDIS, where a pion (or a Kaon) is detected in the final state is presented, in view of constructing a realistic description of the nuclear initial and final states.

  19. Preparation of soft magnetic films of nanocrystalline Fe-Cu-Nb-Si-B alloy by facing targets sputtering

    NASA Astrophysics Data System (ADS)

    Naoe, Masahiko; Matsumiya, Hiroaki; Ichihara, Takayuki; Nakagawa, Shigeki

    1998-06-01

    Soft magnetic thin films of nanocrystalline Fe-Cu-Nb-Si-B alloy were deposited using the facing targets sputtering (FTS) apparatus. It was found that the Fe-Cu-Nb-Si-B single layers thinner than 100 nm revealed good soft magnetic properties, of which the saturation magnetization 4πMs and the relative permeability μr were 11.3 kG and 500, respectively. However, when these films were thicker than 100 nm, their soft magnetic properties degraded due to the perpendicular magnetic anisotropy. On the other hand, the soft magnetic properties of the post-annealed films were improved owing to the release of stress in the films. Especially, μr of the post-annealed films with thickness of 120 nm increased drastically up to around 6200. Furthermore, Fe-Cu-Nb-Si-B/Al multilayers revealed superior soft magnetic properties due to the magnetostatic coupling between the two ferromagnetic layers. These multilayers post-annealed at 300 °C revealed softer magnetic properties than single layers. They exhibited very low coercivity Hc of 0.63 Oe, large 4πMs of 13.2 kG and high μr of 4600.

  20. Retrovirus-mediated siRNA targeting TRPM7 gene induces apoptosis in RBL-2H3 cells.

    PubMed

    Ng, N-M; Jiang, S-P; Lv, Z-Q

    2012-09-01

    Calcium signaling is important for both normal physiologic processes and pathology of various diseases. Transient receptor potential melastatin 7 (TRPM7) gene has been reported to be a potential candidate for calcium influx. The present study aimed to investigate the possible role of TRPM7 channels in apoptosis in rat basophilic leukemia mast cell line (RBL-2H3), which is widely used in mast cell-associated studies. A recombinant retrovirus vector siRNA targeting rat TRPM7 gene was constructed and identified. Cellular survival was assessed by MTT. Cell apoptosis was evaluated by flow cytometry and TUNEL-FITC/Hoechst 33258 staining. The transfection efficiency by retrovirus vector was about 60%-70%. Transfection with TRPM7 siRNA significantly reduced TRPM7 expression both at mRNA and protein levels. Suppression of TRPM7 expression by siRNA led to significantly decreased cellular survival rates and increased apoptosis rates in RBL-2H3 cells. This study indicates that TRPM7 is involved in the apoptosis process in RBL-2H3 cells.

  1. Thermodynamic stability and Watson-Crick base pairing in the seed duplex are major determinants of the efficiency of the siRNA-based off-target effect.

    PubMed

    Ui-Tei, Kumiko; Naito, Yuki; Nishi, Kenji; Juni, Aya; Saigo, Kaoru

    2008-12-01

    Short interfering RNA (siRNA) may down-regulate many unintended genes whose transcripts possess complementarity to the siRNA seed region, which contains 7 nt. The capability of siRNA to induce this off-target effect was highly correlated with the calculated melting temperature or standard free-energy change for formation of protein-free seed duplex, indicating that thermodynamic stability of seed duplex formed between the seed and target is one of the major factor in determining the degree of off-target effects. Furthermore, unlike intended gene silencing (RNA interference), off-target effect was completely abolished by introduction of a G:U pair into the seed duplex, and this loss in activity was completely recovered by a second mutation regenerating Watson-Crick pairing, indicating that seed duplex Watson-Crick pairing is also essential for off-target gene silencing. The off-target effect was more sensitive to siRNA concentration compared to intended gene silencing, which requires a near perfect sequence match between the siRNA guide strand and target mRNA.

  2. In Vitro and In Vivo Tumor-Targeting siRNA Delivery Using Folate-PEG-appended Dendrimer (G4)/α-Cyclodextrin Conjugates.

    PubMed

    Ohyama, Ayumu; Higashi, Taishi; Motoyama, Keiichi; Arima, Hidetoshi

    2016-03-16

    We previously reported that folate-polyethylene glycol (PEG)-appended dendrimer (generation 3)/α-cyclodextrin conjugate (Fol-PαC (G3)) shows folate receptor-α (FR-α)-overexpressing tumor cell-selective in vitro siRNA transfer activity. However, Fol-PαC (G3)/siRNA complex did not induce a significant in vivo RNAi effect after intravenous administration to tumor-bearing mice, possibly resulting from immediate dissociation of the complex in blood. Herein, to develop the novel siRNA carrier having high blood circulating ability, high in vivo siRNA transfer activity, and high safety profile, we newly prepared Fol-PαCs with higher generation (G4) and evaluated their potential as tumor-targeting siRNA carriers in vitro and in vivo. Fol-PαC (G4, average degree of substitution of α-cyclodextrin (DSC) 2.9, average degree of substitution of folate-PEG (DSF) 2)/siRNA complex had the prominent RNAi effect through adequate physicochemical properties, FR-α-mediated endocytosis, efficient endosomal escape, and siRNA delivery to cytoplasm with negligible cytotoxicity. Importantly, Fol-PαC (G4, DSC2.9, DSF2) improved the serum stability, blood circulating ability, and in vivo RNAi effects of siRNA, compared to Fol-PαC (G3). Furthermore, Fol-PαC (G4, DSC2.9, DSF2) complex with siRNA against Polo-like kinase 1 (siPLK1) suppressed the tumor growth compared to control siRNA complex. These results suggest that Fol-PαC (G4, DSC2.9, DSF2) has the potential as a novel tumor-targeting siRNA carrier in vitro and in vivo.

  3. Dual-Targeting Multifuntional Mesoporous Silica Nanocarrier for Codelivery of siRNA and Ursolic Acid to Folate Receptor Overexpressing Cancer Cells.

    PubMed

    Zheng, Guirong; Shen, Yiling; Zhao, Ruirui; Chen, Fan; Zhang, Ying; Xu, Aixiao; Shao, Jingwei

    2017-08-16

    A targeting drug delivery system (TDDS) can selectively deliver antitumor drugs to cancerous parts to improve its anticancer efficacy. Hence, a targeted drug delivery system (UA/siVEGF@MSN-FA) coloading ursolic acid (UA) and vascular endothelial growth factor (VEGF) targeted siRNA (siVEGF) based on mesoporous silica (MSN) nanocarrier modified by a folic acid (FA) molecule was designed and synthesized. The MSN-FA nanoparticles were investigated for shape, diameter, and zeta potential and and by infrared (IR) spectroscopy. FR-overexpressing HeLa cells and FR-negative HepG2 cell lines were used to evaluate the in vitro cellular uptake and the cytotoxicity of MSN-FA nanoparticles. The morphology of HeLa cells transfected with siVEGF@MSN-FA was observed using fluorescence microscopy. Our findings demonstrated that UA@MSN-FA nanoparticles were near-spherical, and the particle size was about 209 ± 9.21 nm. The MSN-FA nanocarrier not only could enhance the in vitro transfection efficiency and the stability of siVEGF but also could further improve the targeted anticancer efficacy of UA and siVEGF via the active targeting property of FA. Overall, the MSN-FA drug delivery system could serve as an excellent material in biomedical applications.

  4. Interleukin-4 receptor-targeted delivery of Bcl-xL siRNA sensitizes tumors to chemotherapy and inhibits tumor growth.

    PubMed

    Guruprasath, Padmanaban; Kim, Jihoon; Gunassekaran, Gowri Rangaswamy; Chi, Lianhua; Kim, Soyoun; Park, Rang-Woon; Kim, Sang-Hyun; Baek, Moon-Chang; Bae, Sang Mun; Kim, Sang-Yeob; Kim, Dong-Kyu; Park, In-Kyu; Kim, Won-Jong; Lee, Byungheon

    2017-10-01

    IL-4 receptor (IL-4R) is commonly up-regulated on tumor cells, and interactions between the receptor and Interleukin-4 (IL-4) can induce the expression of anti-apoptotic proteins, including Bcl-xL. This contributes to tumor cell survival and their resistance to chemotherapy. In this study, we exploited IL-4R-targeted delivery of Bcl-xL siRNA to IL-4R-expressing tumor cells in order to sensitize them to chemotherapy. To target IL-4R, an IL-4R-binding peptide, IL4RPep-1, was attached to branched polyethyleneimine-superparamagnetic iron oxide nanoparticles (BPEI-SPION). These nanoparticles were then complexed with Bcl-xL-targeting siRNA. IL-4R-targeted BPEI-SPION/Bcl-xL siRNA more efficiently reduced Bcl-xL gene expression and enhanced cytotoxicity of doxorubicin in MDA-MB231 breast tumor cells compared to untargeted BPEI-SPION/Bcl-xL siRNA. The siRNA was released from the complexes after 15 h of incubation at pH 5.5 and was stable in the complexes up to 72 h in the serum. The IL-4R-targeted BPEI-SPION/siRNA was internalized by cells through IL-4R, successfully escaped the endosomes, and was dispersed into the cytoplasm. Near-infrared fluorescence and magnetic resonance imaging demonstrated that in vivo tumor homing and accumulation of IL-4R-targeted BPEI-SPION/siRNA were both higher than untargeted BPEI-SPION/siRNA. The IL-4R-targeted BPEI-SPION/Bcl-xL siRNA, in combination with doxorubicin, significantly inhibited tumor growth in mice compared to untargeted BPEI-SPION/Bcl-xL siRNA. These results suggest that the IL-4R-targeted delivery of Bcl-xL siRNA to IL-4R-expressing tumors can sensitize tumors to chemotherapy and enhance the efficacy of anti-tumor therapeutics. Copyright © 2017 Elsevier Ltd. All rights reserved.

  5. Targeting Cell Cycle Proteins in Breast Cancer Cells with siRNA by Using Lipid-Substituted Polyethylenimines

    PubMed Central

    Parmar, Manoj B.; Aliabadi, Hamidreza Montazeri; Mahdipoor, Parvin; Kucharski, Cezary; Maranchuk, Robert; Hugh, Judith C.; Uludağ, Hasan

    2015-01-01

    The cell cycle proteins are key regulators of cell cycle progression whose deregulation is one of the causes of breast cancer. RNA interference (RNAi) is an endogenous mechanism to regulate gene expression and it could serve as the basis of regulating aberrant proteins including cell cycle proteins. Since the delivery of small interfering RNA (siRNA) is a main barrier for implementation of RNAi therapy, we explored the potential of a non-viral delivery system, 2.0 kDa polyethylenimines substituted with linoleic acid and caprylic acid, for this purpose. Using a library of siRNAs against cell cycle proteins, we identified cell division cycle protein 20 (CDC20), a recombinase RAD51, and serine–threonine protein kinase CHEK1 as effective targets for breast cancer therapy, and demonstrated their therapeutic potential in breast cancer MDA-MB-435, MDA-MB-231, and MCF7 cells with respect to another well-studied cell cycle protein, kinesin spindle protein. We also explored the efficacy of dicer-substrate siRNA (DsiRNA) against CDC20, RAD51, and CHEK1, where a particular DsiRNA against CDC20 showed an exceptionally high inhibition of cell growth in vitro. There was no apparent effect of silencing selected cell cycle proteins on the potency of the chemotherapy drug doxorubicin. The efficacy of DsiRNA against CDC20 was subsequently assessed in a xenograft model, which indicated a reduced tumor growth as a result of CDC20 DsiRNA therapy. The presented study highlighted specific cell cycle protein targets critical for breast cancer therapy, and provided a polymeric delivery system for their effective down-regulation. PMID:25763370

  6. Atomistic-continuum modeling of short laser pulse melting of Si targets

    NASA Astrophysics Data System (ADS)

    Lipp, V. P.; Rethfeld, B.; Garcia, M. E.; Ivanov, D. S.

    2014-12-01

    We present an atomistic-continuum model to simulate ultrashort-pulse laser melting processes in semiconductor solids on the example of silicon. The kinetics of transient nonequilibrium phase transition mechanisms is addressed with a molecular dynamics method at atomic level, whereas the laser light absorption, strong generated electron-phonon nonequilibrium, fast diffusion of and heat conduction due to photoexcited free carriers are accounted for in the continuum. We give a detailed description of the model, which is then applied to study the mechanism of short laser pulse melting of freestanding Si films. The effect of laser-induced pressure and temperature of the lattice on the melting kinetics is investigated. Two competing melting mechanisms, heterogeneous and homogeneous, were identified. Apart from the classical heterogeneous melting mechanism, the nucleation of the liquid phase homogeneously inside the material significantly contributes to the melting process. The simulations showed, that due to the open diamond structure of the crystal, the laser-generated internal compressive stresses reduce the crystal stability against the homogeneous melting. Consequently, the latter can take a massive character within several picoseconds upon the laser heating. Due to the negative volume of melting of modeled Si material, -7.5%, the material contracts upon the phase transition, relaxes the compressive stresses, and the subsequent melting proceeds heterogeneously until the excess of thermal energy is consumed. The threshold fluence value, at which homogeneous nucleation of liquid starts contributing to the classical heterogeneous propagation of the solid-liquid interface, is found from the series of simulations at different laser input fluences. For the example of Si, the laser melting kinetics of semiconductors was found to be noticeably different from that of metals with a fcc crystal structure.

  7. Thin-film monocrystalline-silicon solar cells based on a seed layer approach with 11% efficiency

    NASA Astrophysics Data System (ADS)

    Gordon, I.; Qiu, Y.; Van Gestel, D.; Poortmans, J.

    2010-09-01

    Solar modules made from thin-film crystalline-silicon layers of high quality on glass substrates could lower the price of photovoltaic electricity substantially. Almost half of the price of wafer-based silicon solar modules is currently due to the cost of the silicon wafers themselves. Using crystalline-silicon thin-film as the active material would substantially reduce the silicon consumption while still ensuring a high cell-efficiency potential and a stable cell performance. One way to create a crystalline-silicon thin film on glass is by using a seed layer approach in which a thin crystalline-silicon layer is first created on a non-silicon substrate, followed by epitaxial thickening of this layer. In this paper, we present new solar cell results obtained on 10-micron thick monocrystalline-silicon layers, made by epitaxial thickening of thin seed layers on transparent glass-ceramic substrates. We used thin (001)-oriented silicon single-crystal seed layers on glass-ceramic substrates provided by Corning Inc. that are made by a process based on anodic bonding and implant-induced separation. Epitaxial thickening of these seed layers was realized in an atmospheric-pressure chemical vapor deposition system. Simple solar cell structures in substrate configuration were made from the epitaxial mono-silicon layers. The Si surface was plasma-textured to reduce the front-side reflection. No other light trapping features were incorporated. Efficiencies of up to 11% were reached with Voc values above 600 mV indicating the good electronic quality of the material. We believe that by further optimizing the material quality and by integrating an efficient light trapping scheme, the efficiency potential of these single-crystal silicon thin films on glass-ceramics should be higher than 15%.

  8. MDR1 siRNA loaded hyaluronic acid-based CD44 targeted nanoparticle systems circumvent paclitaxel resistance in ovarian cancer

    NASA Astrophysics Data System (ADS)

    Yang, Xiaoqian; Lyer, Arun K.; Singh, Amit; Choy, Edwin; Hornicek, Francis J.; Amiji, Mansoor M.; Duan, Zhenfeng

    2015-02-01

    Development of multidrug resistance (MDR) is an almost universal phenomenon in patients with ovarian cancer, and this severely limits the ultimate success of chemotherapy in the clinic. Overexpression of the MDR1 gene and corresponding P-glycoprotein (Pgp) is one of the best known MDR mechanisms. MDR1 siRNA based strategies were proposed to circumvent MDR, however, systemic, safe, and effective targeted delivery is still a major challenge. Cluster of differentiation 44 (CD44) targeted hyaluronic acid (HA) based nanoparticle has been shown to successfully deliver chemotherapy agents or siRNAs into tumor cells. The goal of this study is to evaluate the ability of HA-PEI/HA-PEG to deliver MDR1 siRNA and the efficacy of the combination of HA-PEI/HA-PEG/MDR1 siRNA with paclitaxel to suppress growth of ovarian cancer. We observed that HA-PEI/HA-PEG nanoparticles can efficiently deliver MDR1 siRNA into MDR ovarian cancer cells, resulting in down-regulation of MDR1 and Pgp expression. Administration of HA-PEI/HA-PEG/MDR1 siRNA nanoparticles followed by paclitaxel treatment induced a significant inhibitory effect on the tumor growth, decreased Pgp expression and increased apoptosis in MDR ovarian cancer mice model. Our findings suggest that CD44 targeted HA-PEI/HA-PEG/MDR1 siRNA nanoparticles can serve as a therapeutic tool with great potentials to circumvent MDR in ovarian cancer.

  9. MDR1 siRNA loaded hyaluronic acid-based CD44 targeted nanoparticle systems circumvent paclitaxel resistance in ovarian cancer

    PubMed Central

    Yang, Xiaoqian; lyer, Arun K.; Singh, Amit; Choy, Edwin; Hornicek, Francis J.; Amiji, Mansoor M.; Duan, Zhenfeng

    2015-01-01

    Development of multidrug resistance (MDR) is an almost universal phenomenon in patients with ovarian cancer, and this severely limits the ultimate success of chemotherapy in the clinic. Overexpression of the MDR1 gene and corresponding P-glycoprotein (Pgp) is one of the best known MDR mechanisms. MDR1 siRNA based strategies were proposed to circumvent MDR, however, systemic, safe, and effective targeted delivery is still a major challenge. Cluster of differentiation 44 (CD44) targeted hyaluronic acid (HA) based nanoparticle has been shown to successfully deliver chemotherapy agents or siRNAs into tumor cells. The goal of this study is to evaluate the ability of HA-PEI/HA-PEG to deliver MDR1 siRNA and the efficacy of the combination of HA-PEI/HA-PEG/MDR1 siRNA with paclitaxel to suppress growth of ovarian cancer. We observed that HA-PEI/HA-PEG nanoparticles can efficiently deliver MDR1 siRNA into MDR ovarian cancer cells, resulting in down-regulation of MDR1 and Pgp expression. Administration of HA-PEI/HA-PEG/MDR1 siRNA nanoparticles followed by paclitaxel treatment induced a significant inhibitory effect on the tumor growth, decreased Pgp expression and increased apoptosis in MDR ovarian cancer mice model. Our findings suggest that CD44 targeted HA-PEI/HA-PEG/MDR1 siRNA nanoparticles can serve as a therapeutic tool with great potentials to circumvent MDR in ovarian cancer. PMID:25687880

  10. Detection of siRNA-mediated target mRNA cleavage activities in human cells by a novel stem-loop array RT-PCR analysis

    PubMed Central

    Lin, Jing; Xu, Kai; Roth, Jack A.; Ji, Lin

    2016-01-01

    The small interfering RNA (siRNA)-mediated target mRNA cleavage activity generates cleaved mRNA fragments with varied termini, which creates major technical challenges for the accurate and efficient detection and verification of cleavage sites on target mRNAs. Here we used a sensitive stem-loop array reverse transcription polymerase chain reaction (SLA-RT-PCR) approach to detect and verify the siRNA-mediated target mRNA cleavage sites by determining precise sequences at the 3′-termini of cleaved mRNA fragments in human cells under physiological conditions. Our results demonstrated the great potential and broad applications of using the SLA-RT-PCR as a sensitive, cost-efficient, and high-throughput tool to systematically detect siRNA-targeted mRNA cleavage sites and fragments in human cells. PMID:26949742

  11. Silencing β3 Integrin by Targeted ECO/siRNA Nanoparticles Inhibits EMT and Metastasis of Triple-Negative Breast Cancer.

    PubMed

    Parvani, Jenny G; Gujrati, Maneesh D; Mack, Margaret A; Schiemann, William P; Lu, Zheng-Rong

    2015-06-01

    Metastatic breast cancer is the second leading cause of cancer-related deaths among women. Triple-negative breast cancer (TNBC) is a highly aggressive subcategory of breast cancer and currently lacks well-defined molecular targets for effective targeted therapies. Disease relapse, metastasis, and drug resistance render standard chemotherapy ineffective in the treatment of TNBC. Because previous studies coupled β3 integrin (ITGB3) to epithelial-mesenchymal transition (EMT) and metastasis, we exploited β3 integrin as a therapeutic target to treat TNBC by delivering β3 integrin siRNA via lipid ECO-based nanoparticles (ECO/siβ3). Treatment of TNBC cells with ECO/siβ3 was sufficient to effectively silence β3 integrin expression, attenuate TGFβ-mediated EMT and invasion, restore TGFβ-mediated cytostasis, and inhibit three-dimensional organoid growth. Modification of ECO/siβ3 nanoparticles with an RGD peptide via a PEG spacer enhanced siRNA uptake by post-EMT cells. Intravenous injections of RGD-targeted ECO/siβ3 nanoparticles in vivo alleviated primary tumor burden and, more importantly, significantly inhibited metastasis. In the span of 16 weeks of the experiments and observations, including primary tumor resection at week 9 and release from the treatment for 4 weeks, the mice bearing orthotopic, TGFβ-prestimulated MDA-MB-231 tumors that were treated with RGD-targeted ECO/siβ3 nanoparticles were free of metastases and relapse, in comparison with untreated mice. Collectively, these results highlight ECO/siβ3 nanoparticles as a promising therapeutic regimen to combat TNBC. ©2015 American Association for Cancer Research.

  12. Therapeutic silence of pleiotrophin by targeted delivery of siRNA and its effect on the inhibition of tumor growth and metastasis

    PubMed Central

    Xie, Weidong; Cheng, Jin; Li, Tong; Mohsen, Mona O.; Lei, Fan; Storni, Federico; Bachmann, Martin; Chen, Hongquan; Zhang, Yaou

    2017-01-01

    Pleiotrophin (PTN) is a secreted cytokine that is expressed in various cancer cell lines and human tumor such as colon cancer, lung cancer, gastric cancer and melanoma. It plays significant roles in angiogenesis, metastasis, differentiation and cell growth. The expression of PTN in the adult is limited to the hippocampus in an activity-dependent manner, making it a very attractive target for cancer therapy. RNA interference (RNAi) offers great potential as a new powerful therapeutic strategy based on its highly specific and efficient silencing of a target gene. However, efficient delivery of small interfering RNA (siRNA) in vivo remains a significant hurdle for its successful therapeutic application. In this study, we first identified, on a cell-based experiment, applying a 1:1 mixture of two PTN specific siRNA engenders a higher silencing efficiency on both mRNA and protein level than using any of them discretely at the same dose. As a consequence, slower melanoma cells growth was also observed for using two specific siRNA combinatorially. To establish a robust way for siRNA delivery in vivo and further investigate how silence of PTN affects tumor growth, we tested three different methods to deliver siRNA in vivo: first non-targeted in-vivo delivery of siRNA via jetPEI; second lung targeted delivery of siRNA via microbubble coated jetPEI; third tumor cell targeted delivery of siRNA via transferrin-polyethylenimine (Tf-PEI). As a result, we found that all three in-vivo siRNAs delivery methods led to an evident inhibition of melanoma growth in non-immune deficiency C57BL/6 mice without a measureable change of ALT and AST activities. Both targeted delivery methods showed more significant curative effect than jetPEI. The lung targeted delivery by microbubble coated jetPEI revealed a comparable therapeutic effect with Tf-PEI, indicating its potential application for target delivery of siRNA in vivo. PMID:28562667

  13. Therapeutic silence of pleiotrophin by targeted delivery of siRNA and its effect on the inhibition of tumor growth and metastasis.

    PubMed

    Zha, Lisha; He, Lichun; Xie, Weidong; Cheng, Jin; Li, Tong; Mohsen, Mona O; Lei, Fan; Storni, Federico; Bachmann, Martin; Chen, Hongquan; Zhang, Yaou

    2017-01-01

    Pleiotrophin (PTN) is a secreted cytokine that is expressed in various cancer cell lines and human tumor such as colon cancer, lung cancer, gastric cancer and melanoma. It plays significant roles in angiogenesis, metastasis, differentiation and cell growth. The expression of PTN in the adult is limited to the hippocampus in an activity-dependent manner, making it a very attractive target for cancer therapy. RNA interference (RNAi) offers great potential as a new powerful therapeutic strategy based on its highly specific and efficient silencing of a target gene. However, efficient delivery of small interfering RNA (siRNA) in vivo remains a significant hurdle for its successful therapeutic application. In this study, we first identified, on a cell-based experiment, applying a 1:1 mixture of two PTN specific siRNA engenders a higher silencing efficiency on both mRNA and protein level than using any of them discretely at the same dose. As a consequence, slower melanoma cells growth was also observed for using two specific siRNA combinatorially. To establish a robust way for siRNA delivery in vivo and further investigate how silence of PTN affects tumor growth, we tested three different methods to deliver siRNA in vivo: first non-targeted in-vivo delivery of siRNA via jetPEI; second lung targeted delivery of siRNA via microbubble coated jetPEI; third tumor cell targeted delivery of siRNA via transferrin-polyethylenimine (Tf-PEI). As a result, we found that all three in-vivo siRNAs delivery methods led to an evident inhibition of melanoma growth in non-immune deficiency C57BL/6 mice without a measureable change of ALT and AST activities. Both targeted delivery methods showed more significant curative effect than jetPEI. The lung targeted delivery by microbubble coated jetPEI revealed a comparable therapeutic effect with Tf-PEI, indicating its potential application for target delivery of siRNA in vivo.

  14. Efficient siRNA Delivery by Lipid Nanoparticles Modified with a Nonstandard Macrocyclic Peptide for EpCAM-Targeting.

    PubMed

    Sakurai, Yu; Mizumura, Wataru; Murata, Manami; Hada, Tomoya; Yamamoto, Shoshiro; Ito, Kenichiro; Iwasaki, Kazuhiro; Katoh, Takayuki; Goto, Yuki; Takagi, Asako; Kohara, Michinori; Suga, Hiroaki; Harashima, Hideyoshi

    2017-08-25

    The development of a specific, effective method for the delivery of therapeutics including small molecules and nucleic acids to tumor tissue remains to be solved. Numerous types of lipid nanoparticles (LNPs) have been developed in attempts to achieve this goal. However, LNPs are probably not taken up by target cells because cancer-targeting LNPs are typically modified with poly(ethylene glycol) (PEG), which inhibits the cellular uptake of LNPs, to passively accumulate in tumor tissue via the enhanced permeability and retention (EPR) effect. It would clearly be important to develop a LNP with both a prolonged circulation and cancer-specific efficient uptake for use in an innovative nanodrug delivery system. Herein, we assessed the effect of nonstandard macrocyclic peptides against the epithelial cell adhesion molecule (EpCAM) Epi-1, which was discovered by a random nonstandard peptides integrated discovery (RaPID) system, on the cellular uptake and therapeutics delivery of LNPs. A liposomal siRNA delivery system (MEND) was modified with an Epi-1 lipid-derivative (EpCAM-targeting MEND; ET-MEND). The resulting ET-MEND showed a more than 27-fold increase in cellular uptake in EpCAM-positive cell lines. In the case of negative cells, cellular uptake and the efficiency of the ET-MEND for delivering therapeutics were comparable with those of nonmodified MEND. In addition, when systemically injected, the ET-MEND successfully inhibited gene expression in the tumor tissue at a dose of 0.5 mg siRNA/kg without any obvious toxicity. These results suggest that a combination of a specific peptide ligand can be used to identify a RaPID system and that the use of such a MEND for liposomal drug delivery has the potential for use in developing a system for the efficacious delivery of pharmaceuticals to various cancer cells.

  15. In vitro study of color stability of polycrystalline and monocrystalline ceramic brackets

    PubMed Central

    de Oliveira, Cibele Braga; Maia, Luiz Guilherme Martins; Santos-Pinto, Ary; Gandini Júnior, Luiz Gonzaga

    2014-01-01

    Objective The aim of this in vitro study was to analyze color stability of monocrystalline and polycrystalline ceramic brackets after immersion in dye solutions. Methods Seven ceramic brackets of four commercial brands were tested: Two monocrystalline and two polycrystalline. The brackets were immersed in four dye solutions (coffee, red wine, Coke and black tea) and in artificial saliva for the following times: 24 hours, 7, 14 and 21 days, respectively. Color changes were measured by a spectrophotometer. Data were assessed by Multivariate Profile Analysis, Analysis of Variance (ANOVA) and Multiple Comparison Tests of means. Results There was a perceptible change of color in all ceramic brackets immersed in coffee (ΔE* Allure = 7.61, Inspire Ice = 6.09, Radiance = 6.69, Transcend = 7.44), black tea (ΔE* Allure = 6.24, Inspire Ice = 5.21, Radiance = 6.51, Transcend = 6.14) and red wine (ΔE* Allure = 6.49, Inspire Ice = 4.76, Radiance = 5.19, Transcend = 5.64), but no change was noticed in Coke and artificial saliva (ΔE < 3.7). Conclusion Ceramic brackets undergo color change when exposed to solutions of coffee, black tea and red wine. However, the same crystalline structure, either monocrystalline or polycrystalline, do not follow the same or a similar pattern in color change, varying according to the bracket fabrication, which shows a lack of standardization in the manufacturing process. Coffee dye produced the most marked color changes after 21 days of immersion for most ceramic brackets evaluated. PMID:25279530

  16. FAST TRACK COMMUNICATION: Synthesis of cubic-structured monocrystalline titanium nitride nanoparticles by means of a dual plasma process

    NASA Astrophysics Data System (ADS)

    Tavares, J.; Coulombe, S.; Meunier, J.-L.

    2009-05-01

    Titanium nitride has long been used for its favourable mechanical and chemical properties and it has been demonstrated that monocrystallinity in thin films enhances these properties. While the synthesis of monocrystalline thin films is well documented, common synthesis processes for titanium nitride nanoparticles yield only polycrystalline, spherically shaped powders. The process presented here allows for the synthesis of monocrystalline, cube-shaped nanoparticles by means of a dual plasma process. Pulsed electric arc erosion of a Ti cathode in a N-rich atmosphere produced by a radio-frequency discharge is used for the synthesis of the TiN nanoparticles. Electron microscopy revealed the cubic morphology of the synthesized powders and electron diffraction patterning confirmed the crystalline structure of the TiN nanoparticles.

  17. Destruction of monocrystalline silicon with nanosecond pulsed fiber laser accompanied by the oxidation of ablation microparticles

    NASA Astrophysics Data System (ADS)

    Veiko, V. P.; Skvortsov, A. M.; Huynh, C. T.; Petrov, A. A.

    2013-11-01

    In this work, we report an observation of process of local destruction monocrystalline silicon with a scanning beam irradiation of pulse ytterbium fiber laser with a wavelength λ= 1062 nm, accompanied by the oxidation of ablation microparticles. It is shown that depending on the power density of irradiation was observed a large scatter size of the microparticles. From a certain average power density is observed beginning oxidation particulate emitted from the surface of the irradiated area. By varying the parameters of the laser beam such as scanning speed, pulse repetition rate, overlap of laser spot, radiation dose can be achieved almost complete oxidation of all formed during the ablation of microparticles.

  18. Folate-targeted amphiphilic cyclodextrin nanoparticles incorporating a fusogenic peptide deliver therapeutic siRNA and inhibit the invasive capacity of 3D prostate cancer tumours.

    PubMed

    Evans, James C; Malhotra, Meenakshi; Sweeney, Katrina; Darcy, Raphael; Nelson, Colleen C; Hollier, Brett G; O'Driscoll, Caitriona M

    2017-09-13

    The main barrier to the development of an effective RNA interference (RNAi) therapy is the lack of a suitable delivery vector. Modified cyclodextrins have emerged in recent years for the delivery of siRNA. In the present study, a folate-targeted amphiphilic cyclodextrin was formulated using DSPE-PEG5000-folate to target prostate cancer cells. The fusogenic peptide GALA was included in the formulation to aid in the endosomal release of siRNA. Targeted nanoparticles were less than 200nm in size with a neutral surface charge. The complexes were able to bind siRNA and protect it from serum nucleases. Incubation with excess free folate resulted in a significant decrease in the uptake of targeted nanoparticles in LNCaP and PC3 cells, both of which have been reported to have differing pathways of folate uptake. There was a significant reduction in the therapeutic targets, ZEB1 and NRP1 at mRNA and protein level following treatment with targeted complexes. In preliminary functional assays using 3D spheroids, treatment of PC3 tumours with targeted complexes with ZEB1 and NRP1 siRNA resulted in more compact colonies relative to the untargeted controls and inhibited infiltration into the Matrigel™ layer. Copyright © 2017 Elsevier B.V. All rights reserved.

  19. Synergistic Suppression of Tumor Angiogenesis by the Co-delivering of Vascular Endothelial Growth Factor Targeted siRNA and Candesartan Mediated by Functionalized Carbon Nanovectors.

    PubMed

    Ding, Xuefang; Su, Yujie; Wang, Cheng; Zhang, Fangrong; Chen, Kerong; Wang, Yu; Li, Min; Wang, Wei

    2017-07-19

    Single-walled carbon nanotubes (SWNTs) with unique physicochemical properties have exhibited promising biomedical applications as drug and gene carriers. In this study, polyethylenimine (PEI)-modified SWNT conjugates linked with candesartan (CD) were developed to deliver vascular endothelial growth factor (VEGF)-targeted siRNA (siVEGF) for the synergistic and targeted treatment of tumor angiogenesis. The characterization results revealed that SWNT-PEI-CD conjugates were successfully synthesized and exhibited desirable dispersibility and superior stability. Confocal laser scanning microscopy (CLSM) and flow cytometry (FCM) results showed that SWNT-PEI-CD/siVEGF complexes could achieve high cellular uptake and specific intracellular distribution of siRNA in AT1R overexpressed PANC-1 cells. Strong down-regulation of VEGF was also verified by qualitative real-time polymerase chain reaction, enzyme-linked immunosorbent assay, and Western blot in complex-treated PANC-1 cells. The in vitro angiogenesis assay showed that SWNT-PEI-CD/siVEGF complexes highly inhibited tube formation of human umbilical vein endothelial cells. Furthermore, in vivo observation in PANC-1 xenografted nude mice demonstrated that SWNT-PEI-CD/siVEGF complexes exhibited significant distribution at tumor sites and caused obvious inhibition of tumor growth and tumor-associated angiogenesis repression induced by the drug combination of CD and siVEGF. Finally, a WST-1 assay indicated that SWNT-PEI-CD possessed low cytotoxicity, and a hemolysis test showed good biocompatibility of SWNT-PEI-CD. Hematological and histological analyses confirmed that SWNT-PEI-CD/siVEGF complexes did not cause any obvious toxic effects to blood and major organs. These findings suggested that the SWNT-PEI-CD/siVEGF co-delivery system with tumor-targeting specificity, improved endosomal escaping properties, and collaboration of angiogenesis inhibition could be a prospective method for efficient tumor antiangiogenic therapy.

  20. Analysis of Laser-Imprinting Reduction in Spherical-RT Experiments with Si-Doped Plastic Targets

    NASA Astrophysics Data System (ADS)

    Hu, S. X.; Fiksel, G.; Goncharov, V. N.; Skupsky, S.

    2011-10-01

    Nonuniformities seeded by both long- and short-wavelength laser perturbations grow during shell implosion as a result of the RT instability, affecting the target performance. To study the effect of high- Z dopants in the ablator material on laser imprint, spherical-RT experiments have been performed at the Omega Laser Facility using Si-doped plastic targets in the cone-in-shell configuration. Compared to the pure plastic target, radiation preheating from the dopant is expected to decrease the mass density at the ablation front and increase the stand-off distance between the ablation front and laser-deposition region, reducing the imprinting efficiency and the RT growth. Analyses of experimental data using two-dimensional DRACO simulations for cases with and without dopants will be presented. This work was supported by the U.S. Department of Energy Office of Inertial Confinement Fusion under Cooperative Agreement No. DE-FC52-08NA28302. V. N. Goncharovet al., Phys. Plasmas 10, 1906 (2003).

  1. On the efficiency calibration of Si(Li) detector in the low-energy region using thick-target bremsstrahlung

    NASA Astrophysics Data System (ADS)

    An, Z.; Liu, M. T.

    2002-10-01

    In this paper, the efficiency calibration of a Si(Li) detector in the low-energy region down to 0.58 keV has been performed using thick-carbon-target bremsstrahlung by 19 keV electron impact. The shape of the efficiency calibration curve was determined from the thick-carbon-target bremsstrahlung spectrum, and the absolute value for the efficiency calibration was obtained from the use of 241Am radioactive standard source. The modified Wentzel's formula for thick-target bremsstrahlung was employed and it was also compared with the most recently developed theoretical model based upon the doubly differential cross-sections for bremsstrahlung of Kissel, Quarles and Pratt. In the present calculation of theoretical bremsstrahlung, the self-absorption correction and the convolution of detector's response function with the bremsstrahlung spectrum have simultaneously been taken into account. The accuracy for the efficiency calibration in the low-energy region with the method described here was estimated to be about 6%. Moreover, the self-absorption correction calculation based upon the prescription of Wolters et al. has also been presented as an analytical factor with the accuracy of ˜1%.

  2. A linear polyethylenimine mediated siRNA-based therapy targeting human epidermal growth factor receptor in SPC-A1 xenograft mice.

    PubMed

    Zhang, Pinghai; Xu, Nuo; Zhou, Lei; Xu, Xin; Wang, Yuehong; Li, Ka; Zeng, Zhaochong; Wang, Xiangdong; Zhang, Xin; Bai, Chunxue

    2013-12-01

    Linear polyethylenimine (LPEI) is considered as a desirable gene in vivo delivery system, but whether it could deliver the specific siRNA targeted EGFR to the tumor site to inhibit the growth of NSCLC xenograft in nude mice still needs to be examined. In this study, LPEI/siRNA was made into a complex and SPC-A1-xenografted mice model was established. Then, stable LPEI/siRNA-EGFR complexes were intraperitoneally administrated. Afterwards, tumor growth was measured every 3 days. At the end of the experiment, tumor volume was calculated, and tumors were weighed, and examined for EGFR expression, proliferation, and apoptosis evaluations. By using blood samples, toxicity tests including aspartate aminotransferase (AST), alanine aminotransferase (ALT), urea and creatinine (Cr) were measured for liver and renal function evaluation. Serum concentrations of TNF-α and IFN-γ were also examined. Our results demonstrated that LPEI/siRNA-EGFR complexes could downregulate EGFR expression in SPC-A1 xenografted tumor upon single i.p. injection. LPEI/siRNA-EGFR complexes inhibited tumor growth and did not induce organ toxicity in SPC-A1-xenografted mice. At the end of the experiment no significant IFN-α increase was detected in LPEI/siRNA complexes or glucose-treated groups. The novel modality of siRNA-based therapy targeting EGFR may be of great potential in NSCLC treatment.

  3. Endothelin system in oral squamous carcinoma cells: specific siRNA targeting of ECE-1 blocks cell proliferation.

    PubMed

    Awano, Shuji; Dawson, Louise A; Hunter, Alison R; Turner, Anthony J; Usmani, Badar A

    2006-04-01

    The present study focused on the endothelin axis in human oral squamous cell carcinoma (SCC) cells. We investigated the expression and distribution of endothelin-1 (ET-1), its receptors (endothelin-A receptor (ET(A)R) and endothelin-B receptor (ET(B)R)) and isoforms of its specific converting enzyme (ECE-1a, 1b, 1c) and the report on their relative influences on cell proliferation. We also investigated the effect of an ECE-specific inhibitor (ECE-i) and siRNA targeting of the ECE-1 gene on SCC cell proliferation. We observed the expression of ET-1, ET(A)R, ET(B)R and all endothelin-converting enzyme-1 (ECE-1) isoforms in oral SCC cells, but only the expression of ET-1, ET(B)R and ECE-1 was increased when compared to normal human epidermal keratinocytes. ET-1 alone stimulated proliferation of oral SCC cells. Antagonists of either ET(A)R or ET(B)R inhibited ET-1-mediated proliferation. Decreased ECE-1 expression after ECE siRNA treatment reduced SCC cell proliferation. Antiproliferative effects were also observed by inhibiting ECE activity with ECE-i. In conclusion, the present study demonstrates that modulation of the endothelin system in oral SCC cells might provide a novel therapeutic protocol for oral cancer.

  4. 5-ASA-loaded SiO2 nanoparticles-a novel drug delivery system targeting therapy on ulcerative colitis in mice

    PubMed Central

    Tang, Haiying; Xiang, Dan; Wang, Feng; Mao, Jingwei; Tan, Xiaoyan; Wang, Yingde

    2017-01-01

    The targeting of 5-aminosalicylic acid (5-ASA), a first-line therapeutic agent for mild to moderate active ulcerative colitis (UC), to the site of inflammation has remained a challenge and an unmet requirement in the treatment of UC. However, nanoscale carriers for targeted drug delivery are promising for pharmacotherapy, and nanoparticles improve the pharmacokinetics of the loaded therapeutics based on their physical properties. To design and prepare 5-ASA-loaded silicon dioxide nanoparticles (5-ASA-SiO2 NPs), a micro-emulsion method was conducted, and their respective therapeutic effects were validated in a mouse model of UC. Cytotoxicity of 5-ASA-SiO2 NPs was detected in vitro using the Cell Counting Kit-8 method. The therapeutic effect of 5-ASA-SiO2 NPs was assessed based on their disease activity index (DAI), colon histopathology, myeloperoxidase (MPO) and levels of tumor necrosis factor-α (TNF-α) and interleukin-6 (IL-6). SiO2 NPs were successfully prepared, and cytotoxicity of 5-ASA-SiO2 NPs was identified as being similar to 5-ASA and SiO2 NPs. DAI and colonic histopathology scores in the normal dosage, high dosage and the 5-ASA-SiO2 NP groups demonstrated a significant improvement when compared with the model group. DAI in the high dosage and 5-ASA-SiO2 NP groups also demonstrated a significant improvement when compared with the normal dosage group. However, MPO, serum IL-6 and TNF-α levels in normal dosage, high dosage and 5-ASA-SiO2 NPs groups were significantly lower than in the model group, and these indexes in the high dosage group and 5-ASA-SiO2 NP group were significantly lower than that in the normal dosage group. Expression of IL-6 and TNF-α mRNA in colonic mucosa in the normal dosage, high dosage and 5-ASA-SiO2 NP group was significantly lower than that in the model group. Colonic mucosal IL-6 and TNF-α mRNA expression in the high dosage and 5-ASA-SiO2 NP groups was significantly lower than that in the normal dosage group (P<0.05). In

  5. Lipid nanoparticles for targeted siRNA delivery – going from bench to bedside

    PubMed Central

    Zatsepin, Timofei S; Kotelevtsev, Yuri V; Koteliansky, Victor

    2016-01-01

    This review covers the basic aspects of small interfering RNA delivery by lipid nano-particles (LNPs) and elaborates on the current status of clinical trials for these systems. We briefly describe the roles of all LNP components and possible strategies for their improvement. We also focus on the current clinical trials using LNP-formulated RNA and the possible outcomes for therapy in the near future. Also, we present a critical analysis of selected clinical trials that reveals the common logic behind target selection. We address this review to a wide audience, especially to medical doctors who are interested in the application of RNA interference–based treatment platforms. We anticipate that this review may spark interest in this particular audience and generate new ideas in target selection for the disorders they are dealing with. PMID:27462152

  6. Nano-siRNA Particles and Combination Therapies for Ovarian Tumor Targeting

    DTIC Science & Technology

    2015-08-01

    reaction parameters. Treating the multi-RNAi-MS particles with different polycations, such as poly-L-lysine ( PLL ) and polyethylene imine (PEI), yielded...suppression and systemic toxicity. PLL /HA functionalized liposome nanoparticles with ratiometric drug loaded combination therapy will selectively target...therapeutic efficacy. Cells were treated with increasing concentration of both free (A,C,D,E) and liposome PLL /HA drug encapsulated nanoparticles (B). Cells

  7. Nanoencapsulated anti-CK2 small molecule drug or siRNA specifically targets malignant cancer but not benign cells.

    PubMed

    Trembley, Janeen H; Unger, Gretchen M; Korman, Vicci L; Tobolt, Diane K; Kazimierczuk, Zygmunt; Pinna, Lorenzo A; Kren, Betsy T; Ahmed, Khalil

    2012-02-01

    CK2, a pleiotropic Ser/Thr kinase, is an important target for cancer therapy. We tested our novel tenfibgen-based nanocapsule for delivery of the inhibitor 2-dimethylamino-4,5,6,7-tetrabromo-1H-benzimidazole (DMAT) and an siRNA directed against both CK2α and α' catalytic subunits to prostate cancer cells. We present data on the TBG nanocapsule itself and on CK2 inhibition or downregulation in treated cells, including effects on Nuclear Factor-kappa B (NF-κB) p65. By direct comparison of two CK2-directed cargos, our data provide proof that the TBG encapsulation design for delivery of drugs specifically to cancer cells has strong potential for small molecule- and nucleic acid-based cancer therapy.

  8. Enhanced delivery of PEAL nanoparticles with ultrasound targeted microbubble destruction mediated siRNA transfection in human MCF-7/S and MCF-7/ADR cells in vitro

    PubMed Central

    Teng, Yanwei; Bai, Min; Sun, Ying; Wang, Qi; Li, Fan; Xing, Jinfang; Du, Lianfang; Gong, Tao; Duan, Yourong

    2015-01-01

    The gene knockdown activity of small interfering RNA (siRNA) has led to their use as potential therapeutics for a variety of diseases. However, successful gene therapy requires safe and efficient delivery systems. In this study, we choose mPEG-PLGA-PLL nanoparticles (PEAL NPs) with ultrasound targeted microbubble destruction (UTMD) to efficiently deliver siRNA into cells. An emulsification-solvent evaporation method was used to prepare siRNA-loaded PEAL NPs. The NPs possessed an average size of 132.6±10.3 nm (n=5), with a uniform spherical shape, and had an encapsulation efficiency (EE) of more than 98%. As demonstrated by MTT assay, neither PEAL NPs nor siRNA-loaded PEAL NPs showed cytotoxicity even at high concentrations. The results of cellular uptake showed, with the assistance of UTMD, the siRNA-loaded PEAL NPs can be effectively internalized and can subsequently release siRNA in cells. Taken together, PEAL NPs with UTMD may be highly promising for siRNA delivery, making it possible to fully exploit the potential of siRNA-based therapeutics. PMID:26346350

  9. Targeted Delivery of siRNA to Activated T Cells via Transferrin-Polyethylenimine (Tf-PEI) as a Potential Therapy of Asthma

    PubMed Central

    Xie, Yuran; Kim, Na Hyung; Nadithe, Venkatareddy; Schalk, Dana; Thakur, Archana; Kılıç, Ayşe; Lum, Lawrence G.; Bassett, David JP; Merkel, Olivia M

    2016-01-01

    Asthma is a worldwide health problem. Activated T cells (ATCs) in the lung, particularly T helper 2 cells (Th2), are strongly associated with inducing airway inflammatory responses and chemoattraction of inflammatory cells in asthma. Small interfering RNA (siRNA) as a promising anti-sense molecule can specifically silence inflammation related genes in ATCs, however, lack of safe and efficient siRNA delivery systems limits the application of siRNA as a therapeutic molecule in asthma. Here, we designed a novel pulmonary delivery system of siRNA, transferrin-polyethylenimine (Tf-PEI), to selectively deliver siRNA to ATCs in the lung. Tf-PEI polyplexes demonstrated optimal physicochemical properties such as size, distribution, zeta-potential, and siRNA condensation efficiency. Moreover, in vitro studies showed significantly enhanced cellular uptake and gene knockdown mediated by Tf-PEI polyplexes in human primary ATCs. Biodistribution of polyplexes in a murine asthmatic model confirmed that Tf-PEI polyplexes can efficiently and selectively deliver siRNA to ATCs. In conclusion, the present work proves the feasibility to target ATCs in asthma via Tf receptor. This strategy could potentially be used to design an efficient siRNA delivery system for asthma therapy. PMID:27001893

  10. RIsearch2: suffix array-based large-scale prediction of RNA-RNA interactions and siRNA off-targets.

    PubMed

    Alkan, Ferhat; Wenzel, Anne; Palasca, Oana; Kerpedjiev, Peter; Rudebeck, Anders Frost; Stadler, Peter F; Hofacker, Ivo L; Gorodkin, Jan

    2017-01-20

    Intermolecular interactions of ncRNAs are at the core of gene regulation events, and identifying the full map of these interactions bears crucial importance for ncRNA functional studies. It is known that RNA-RNA interactions are built up by complementary base pairings between interacting RNAs and high level of complementarity between two RNA sequences is a powerful predictor of such interactions. Here, we present RIsearch2, a large-scale RNA-RNA interaction prediction tool that enables quick localization of potential near-complementary RNA-RNA interactions between given query and target sequences. In contrast to previous heuristics which either search for exact matches while including G-U wobble pairs or employ simplified energy models, we present a novel approach using a single integrated seed-and-extend framework based on suffix arrays. RIsearch2 enables fast discovery of candidate RNA-RNA interactions on genome/transcriptome-wide scale. We furthermore present an siRNA off-target discovery pipeline that not only predicts the off-target transcripts but also computes the off-targeting potential of a given siRNA. This is achieved by combining genome-wide RIsearch2 predictions with target site accessibilities and transcript abundance estimates. We show that this pipeline accurately predicts siRNA off-target interactions and enables off-targeting potential comparisons between different siRNA designs. RIsearch2 and the siRNA off-target discovery pipeline are available as stand-alone software packages from http://rth.dk/resources/risearch.

  11. Large three-dimensional photonic crystals based on monocrystalline liquid crystal blue phases.

    PubMed

    Chen, Chun-Wei; Hou, Chien-Tsung; Li, Cheng-Chang; Jau, Hung-Chang; Wang, Chun-Ta; Hong, Ching-Lang; Guo, Duan-Yi; Wang, Cheng-Yu; Chiang, Sheng-Ping; Bunning, Timothy J; Khoo, Iam-Choon; Lin, Tsung-Hsien

    2017-09-28

    Although there have been intense efforts to fabricate large three-dimensional photonic crystals in order to realize their full potential, the technologies developed so far are still beset with various material processing and cost issues. Conventional top-down fabrications are costly and time-consuming, whereas natural self-assembly and bottom-up fabrications often result in high defect density and limited dimensions. Here we report the fabrication of extraordinarily large monocrystalline photonic crystals by controlling the self-assembly processes which occur in unique phases of liquid crystals that exhibit three-dimensional photonic-crystalline properties called liquid-crystal blue phases. In particular, we have developed a gradient-temperature technique that enables three-dimensional photonic crystals to grow to lateral dimensions of ~1 cm (~30,000 of unit cells) and thickness of ~100 μm (~ 300 unit cells). These giant single crystals exhibit extraordinarily sharp photonic bandgaps with high reflectivity, long-range periodicity in all dimensions and well-defined lattice orientation.Conventional fabrication approaches for large-size three-dimensional photonic crystals are problematic. By properly controlling the self-assembly processes, the authors report the fabrication of monocrystalline blue phase liquid crystals that exhibit three-dimensional photonic-crystalline properties.

  12. Solution-Phase Epitaxial Growth of Quasi-Monocrystalline Cuprous Oxide on Metal Nanowires

    PubMed Central

    2014-01-01

    The epitaxial growth of monocrystalline semiconductors on metal nanostructures is interesting from both fundamental and applied perspectives. The realization of nanostructures with excellent interfaces and material properties that also have controlled optical resonances can be very challenging. Here we report the synthesis and characterization of metal–semiconductor core–shell nanowires. We demonstrate a solution-phase route to obtain stable core–shell metal–Cu2O nanowires with outstanding control over the resulting structure, in which the noble metal nanowire is used as the nucleation site for epitaxial growth of quasi-monocrystalline Cu2O shells at room temperature in aqueous solution. We use X-ray and electron diffraction, high-resolution transmission electron microscopy, energy dispersive X-ray spectroscopy, photoluminescence spectroscopy, and absorption spectroscopy, as well as density functional theory calculations, to characterize the core–shell nanowires and verify their structure. Metal–semiconductor core–shell nanowires offer several potential advantages over thin film and traditional nanowire architectures as building blocks for photovoltaics, including efficient carrier collection in radial nanowire junctions and strong optical resonances that can be tuned to maximize absorption. PMID:25233392

  13. Solution-phase epitaxial growth of quasi-monocrystalline cuprous oxide on metal nanowires.

    PubMed

    Sciacca, Beniamino; Mann, Sander A; Tichelaar, Frans D; Zandbergen, Henny W; van Huis, Marijn A; Garnett, Erik C

    2014-10-08

    The epitaxial growth of monocrystalline semiconductors on metal nanostructures is interesting from both fundamental and applied perspectives. The realization of nanostructures with excellent interfaces and material properties that also have controlled optical resonances can be very challenging. Here we report the synthesis and characterization of metal-semiconductor core-shell nanowires. We demonstrate a solution-phase route to obtain stable core-shell metal-Cu2O nanowires with outstanding control over the resulting structure, in which the noble metal nanowire is used as the nucleation site for epitaxial growth of quasi-monocrystalline Cu2O shells at room temperature in aqueous solution. We use X-ray and electron diffraction, high-resolution transmission electron microscopy, energy dispersive X-ray spectroscopy, photoluminescence spectroscopy, and absorption spectroscopy, as well as density functional theory calculations, to characterize the core-shell nanowires and verify their structure. Metal-semiconductor core-shell nanowires offer several potential advantages over thin film and traditional nanowire architectures as building blocks for photovoltaics, including efficient carrier collection in radial nanowire junctions and strong optical resonances that can be tuned to maximize absorption.

  14. Controlled Growth of Monocrystalline Organo-Lead Halide Perovskite and Its Application in Photonic Devices.

    PubMed

    Mao, Wenxin; Zheng, Jialu; Zhang, Yupeng; Chesman, Anthony S R; Ou, Qingdong; Hicks, Jamie; Li, Feng; Wang, Ziyu; Graystone, Brenton; Bell, Toby D M; Rothmann, Mathias Uller; Duffy, Noel W; Spiccia, Leone; Cheng, Yi-Bing; Bao, Qiaoliang; Bach, Udo

    2017-10-02

    Organo-lead halide perovskites (OHPs) have recently emerged as a new class of exceptional optoelectronic materials, which may find use in many applications, including solar cells, light emitting diodes, and photodetectors. More complex applications, such as lasers and electro-optic modulators, require the use of monocrystalline perovskite materials to reach their ultimate performance levels. Conventional methods for forming single crystals of OHPs like methylammonium lead bromide (MAPbBr3 ) afford limited control over the product morphology, rendering the assembly of defined microcavity nanostructures difficult. We overcame this by synthesizing for the first time (MA)[PbBr3 ]⋅DMF (1), and demonstrating its facile transformation into monocrystalline MAPbBr3 microplatelets. The MAPbBr3 microplatelets were tailored into waveguide based photonic devices, of which an ultra-low propagation loss of 0.04 dB μm(-1) for a propagation distance of 100 μm was demonstrated. An efficient active electro-optical modulator (AEOM) consisting of a MAPbBr3 non-linear arc waveguide was demonstrated, exhibiting a 98.4 % PL intensity modulation with an external voltage of 45 V. This novel synthetic approach, as well as the demonstration of effective waveguiding, will pave the way for developing a wide range of photonic devices based on organo-lead halide perovskites. © 2017 Wiley-VCH Verlag GmbH & Co. KGaA, Weinheim.

  15. Enhanced cooling in mono-crystalline ultra-thin silicon by embedded micro-air channels

    NASA Astrophysics Data System (ADS)

    Ghoneim, Mohamed T.; Fahad, Hossain M.; Hussain, Aftab M.; Rojas, Jhonathan P.; Torres Sevilla, Galo A.; Alfaraj, Nasir; Lizardo, Ernesto B.; Hussain, Muhammad M.

    2015-12-01

    In today's digital world, complementary metal oxide semiconductor (CMOS) technology enabled scaling of bulk mono-crystalline silicon (100) based electronics has resulted in their higher performance but with increased dynamic and off-state power consumption. Such trade-off has caused excessive heat generation which eventually drains the charge of battery in portable devices. The traditional solution utilizing off-chip fans and heat sinks used for heat management make the whole system bulky and less mobile. Here we show, an enhanced cooling phenomenon in ultra-thin (>10 μm) mono-crystalline (100) silicon (detached from bulk substrate) by utilizing deterministic pattern of porous network of vertical "through silicon" micro-air channels that offer remarkable heat and weight management for ultra-mobile electronics, in a cost effective way with 20× reduction in substrate weight and a 12% lower maximum temperature at sustained loads. We also show the effectiveness of this event in functional MOS field effect transistors (MOSFETs) with high-κ/metal gate stacks.

  16. Task-dependent modulation of SI physiological responses to targets and distractors.

    PubMed

    Spingath, Elsie; Kang, Hyun-Sug; Blake, David T

    2013-02-01

    Selective attention experimental designs have shown that neural responses to stimuli in primary somatosensory cortex are stronger when the sensory stimuli are task relevant. Other studies have used animals under no task demands for data collection. The relationship between neural responses in the brain during behavior, and while an animal has no task demands, remains underexplored. We trained two animals to perform somatosensory detection for several weeks, followed by somatosensory discrimination for several weeks. Data in response to physically identical stimuli were collected from cortical implants while the animal was under no task demands before each behavioral session and also during that behavioral session. The Fourier spectra of the field potentials during detection or discrimination compared with the no task condition demonstrated suppression of the somatosensory μ-rhythm that is associated with readiness and anticipation of cognitive use of somatosensory and motor inputs. Responses to the task target were stronger during detection and discrimination than in the no task condition. The amplitude normalized time course of the target evoked response was similar in both cases. Evoked responses to the task distractor were not significantly stronger during behavior than in recordings under no task demands. The normalized time course of the distractor responses showed a suppression that peaks 30-35 ms after the onset of the response. The selectivity of this within trial suppression is the same as the selectivity of enduring suppression evident in studies of sensory cortical plasticity, which suggests the same neural process may be responsible for both.

  17. Intracellular siRNA delivery dynamics of integrin-targeted, PEGylated chitosan-poly(ethylene imine) hybrid nanoparticles: A mechanistic insight.

    PubMed

    Ragelle, Héloïse; Colombo, Stefano; Pourcelle, Vincent; Vanvarenberg, Kevin; Vandermeulen, Gaëlle; Bouzin, Caroline; Marchand-Brynaert, Jacqueline; Feron, Olivier; Foged, Camilla; Préat, Véronique

    2015-08-10

    Integrin-targeted nanoparticles are promising for the delivery of small interfering RNA (siRNA) to tumor cells or tumor endothelium in cancer therapy aiming at silencing genes essential for tumor growth. However, during the process of optimizing and realizing their full potential, it is pertinent to gain a basic mechanistic understanding of the bottlenecks existing for nanoparticle-mediated intracellular delivery. We designed αvβ3 integrin-targeted nanoparticles by coupling arginine-glycine-aspartate (RGD) or RGD peptidomimetic (RGDp) ligands to the surface of poly(ethylene glycol) (PEG) grafted chitosan-poly(ethylene imine) hybrid nanoparticles. The amount of intracellular siRNA delivered by αvβ3-targeted versus non-targeted nanoparticles was quantified in the human non-small cell lung carcinoma cell line H1299 expressing enhanced green fluorescent protein (EGFP) using a stem-loop reverse transcription quantitative polymerase chain reaction (RT-qPCR) approach. Data demonstrated that the internalization of αvβ3-targeted nanoparticles was highly dependent on the surface concentration of the ligand. Above a certain threshold concentration, the use of targeted nanoparticles provided a two-fold increase in the number of siRNA copies/cell, subsequently resulting in as much as 90% silencing of EGFP at well-tolerated carrier concentrations. In contrast, non-targeted nanoparticles mediated low levels of gene silencing, despite relatively high intracellular siRNA concentrations, indicating that these nanoparticles might end up in late endosomes or lysosomes without releasing their cargo to the cell cytoplasm. Thus, the silencing efficiency of the chitosan-based nanoparticles is strongly dependent on the uptake and the intracellular trafficking in H1299 EGFP cells, which is critical information towards a more complete understanding of the delivery mechanism that can facilitate the future design of efficient siRNA delivery systems.

  18. Dynamic Contrast Enhanced MRI Assessing the Antiangiogenic Effect of Silencing HIF-1α with Targeted Multifunctional ECO/siRNA Nanoparticles.

    PubMed

    Malamas, Anthony S; Jin, Erlei; Gujrati, Maneesh; Lu, Zheng-Rong

    2016-07-05

    Stabilization of hypoxia inducible factor 1α (HIF-1α), a biomarker of hypoxia, in hypoxic tumors mediates a variety of downstream genes promoting tumor angiogenesis and cancer cell survival as well as invasion, and compromising therapeutic outcome. In this study, dynamic contrast enhanced MRI (DCE-MRI) with a biodegradable macromolecular MRI contrast agent was used to noninvasively assess the antiangiogenic effect of RGD-targeted multifunctional lipid ECO/siHIF-1α nanoparticles in a mouse HT29 colon cancer model. The RGD-targeted ECO/siHIF-1α nanoparticles resulted in over 50% reduction in tumor size after intravenous injection at a dose of 2.0 mg of siRNA/kg every 3 days for 3 weeks compared to a saline control. DCE-MRI revealed significant decline in vascularity and over a 70% reduction in the tumor blood flow, permeability-surface area product, and plasma volume fraction vascular parameters in the tumor treated with the targeted ECO/siHIF-1α nanoparticles. The treatment with targeted ECO/siRNA nanoparticles resulted in significant silencing of HIF-1α expression at the protein level, which also significantly suppressed the expression of VEGF, Glut-1, HKII, PDK-1, LDHA, and CAIX, which are all important players in tumor angiogenesis, glycolytic metabolism, and pH regulation. By possessing the ability to elicit a multifaceted effect on tumor biology, silencing HIF-1α with RGD-targeted ECO/siHIF-1α nanoparticles has great promise as a single therapy or in combination with traditional chemotherapy or radiation strategies to improve cancer treatment.

  19. Preparation and characterization of polymeric nanoparticles for siRNA delivery to down-regulate the expressions of exogenous and endogenous target genes.

    PubMed

    Huang, Wei; Lv, Ming; Gao, Zhong-Gao; Jin, Ming-Ji; Xu, Yuan-Ji; Yu, Xiao-Dan; Jin, Zhe-Hu; Yin, Xue-Zhe

    2012-08-01

    Gene silencing induced by RNA interference using small interfering RNA (siRNA) provides a promising therapeutic approach for cancers. However, the lack of siRNA delivery vector has limited the development of siRNA therapy. The purpose of this study was to use the novel copolymer (mPEG5k-PCL1.2k)1.4-g-PEl10k to prepare siRNA-loaded nanoparticles for siRNA delivery. The results suggested that (mPEG5k-PCL1.2k)1.4-g-PEl10k could load siRNA to form nanoparticles with particle size less than 200 nm in a narrow distribution. Moreover, a certain density of positive charge existed onto the surfaces of nanoparticles. MTT assay results demonstrated that (mPEG5k-PCL1.2k)1.4-g-PEl10k/siRNA nanoparticles showed very low cytotoxicity. The gene silencing efficiency of (mPEG5k-PCL1.2k)1.4-g-PEl10k/siRNA nanoparticles was investigated through luciferase reporter gene assays. The expression of exogenous luciferase gene was significantly downregulated at a range of N/P ratio from 50 to 125, and was maximally inhibited at the N/P ratio of 125 with 54% and 59% reduction in MCF-7 and HepG2 cells, respectively. In the 4T1-luc cell line expressing luciferase stably, the silencing of endogenous luciferase gene also has a similar overall profile with maximal 54% reduction of luciferase expression. These results suggested that (mPEG5k-PCL1.2k)1.4-g-PEI10k/SiRNA nanoparticles could serve as a kind of highly efficient siRNA delivery system for down-regulating the expression of exogenous and endogenous target genes.

  20. SKI2 mediates degradation of RISC 5′-cleavage fragments and prevents secondary siRNA production from miRNA targets in Arabidopsis

    PubMed Central

    Branscheid, Anja; Marchais, Antonin; Schott, Gregory; Lange, Heike; Gagliardi, Dominique; Andersen, Stig Uggerhøj; Voinnet, Olivier; Brodersen, Peter

    2015-01-01

    Small regulatory RNAs are fundamental in eukaryotic and prokaryotic gene regulation. In plants, an important element of post-transcriptional control is effected by 20–24 nt microRNAs (miRNAs) and short interfering RNAs (siRNAs) bound to the ARGONAUTE1 (AGO1) protein in an RNA induced silencing complex (RISC). AGO1 may cleave target mRNAs with small RNA complementarity, but the fate of the resulting cleavage fragments remains incompletely understood. Here, we show that SKI2, SKI3 and SKI8, subunits of a cytoplasmic cofactor of the RNA exosome, are required for degradation of RISC 5′, but not 3′-cleavage fragments in Arabidopsis. In the absence of SKI2 activity, many miRNA targets produce siRNAs via the RNA-dependent RNA polymerase 6 (RDR6) pathway. These siRNAs are low-abundant, and map close to the cleavage site. In most cases, siRNAs were produced 5′ to the cleavage site, but several examples of 3′-spreading were also identified. These observations suggest that siRNAs do not simply derive from RDR6 action on stable 5′-cleavage fragments and hence that SKI2 has a direct role in limiting secondary siRNA production in addition to its function in mediating degradation of 5′-cleavage fragments. PMID:26464441

  1. Measurement of proton induced thick target γ-ray yields on B, N, Na, Al and Si from 2.5 to 4.1 MeV

    NASA Astrophysics Data System (ADS)

    Chiari, M.; Ferraccioli, G.; Melon, B.; Nannini, A.; Perego, A.; Salvestrini, L.; Lagoyannis, A.; Preketes-Sigalas, K.

    2016-01-01

    Thick target yields for proton induced γ-ray emission (PIGE) on low-Z nuclei, namely B, N, Na, Al and Si, were measured for proton energies from 2.5 to 4.1 MeV and emission angles of 0°, 45° and 90°, at the 3 MV Tandetron laboratory of INFN-LABEC in Florence. The studied reactions were: 10B(p,α‧γ)7Be (Eγ = 429 keV), 10B(p,p‧γ)10B (Eγ = 718 keV) and 11B(p,p‧γ)11B (Eγ = 2125 keV) for boron; 14N(p,p‧γ)14N (Eγ = 2313 keV) for nitrogen; 23Na(p,p‧γ)23Na (Eγ = 441 and 1636 keV) and 23Na(p,α‧γ)20Ne (Eγ = 1634 keV) for sodium; 27Al(p,p‧γ)27Al (Eγ = 844 and 1014 keV) and 27Al(p,α‧γ)24Mg (Eγ = 1369 keV) for aluminum; 28Si(p,p‧γ)28Si (Eγ = 1779 keV) and 29Si(p,p‧γ)29Si (Eγ = 1273 keV) for silicon. The PIGE thick target yields have been measured with an overall uncertainty typically better than 10%. The use of the measured thick target yield to benchmark and validate experimental cross sections available in the literature is demonstrated.

  2. Ultrasound-assisted siRNA delivery via arginine-grafted bioreducible polymer and microbubbles targeting VEGF for ovarian cancer treatment.

    PubMed

    Florinas, Stelios; Kim, Jaesung; Nam, Kihoon; Janát-Amsbury, Margit M; Kim, Sung Wan

    2014-06-10

    The major drawback hampering siRNA therapies from being more widely accepted in clinical practice is its insufficient accumulation at the target site mainly due to poor cellular uptake and rapid degradation in serum. Therefore, we designed a novel polymeric siRNA carrier system, which would withstand serum-containing environments and tested its performance in vitro as well as in vivo. Delivering siRNA with a system combining an arginine-grafted bioreducible polymer (ABP), microbubbles (MBs), and ultrasound technology (US) we were able to synergize the advantages each delivery system owns individually, and created our innovative siRNA-ABP-MB (SAM) complexes. SAM complexes show significantly higher siRNA uptake and VEGF protein knockdown in vitro with serum-containing media when compared to naked siRNA, and 25k-branched-polyethylenimine (bPEI) representing the current standard in nonviral gene therapy. SAM complexes activated by US are also able to improve siRNA uptake in tumor tissue resulting in decelerating tumor growth in vivo. Copyright © 2014 Elsevier B.V. All rights reserved.

  3. Ovarian cancer immunotherapy using PD-L1 siRNA targeted delivery from folic acid-functionalized polyethylenimine: strategies to enhance T cell killing.

    PubMed

    Teo, Pei Yun; Yang, Chuan; Whilding, Lynsey M; Parente-Pereira, Ana C; Maher, John; George, Andrew J T; Hedrick, James L; Yang, Yi Yan; Ghaem-Maghami, Sadaf

    2015-06-03

    Adoptive T cell immunotherapy is a promising treatment strategy for epithelial ovarian cancer (EOC). However, programmed death ligand-1 (PD-L1), highly expressed on EOC cells, interacts with programmed death-1 (PD-1), expressed on T cells, causing immunosuppression. This study aims to block PD-1/PD-L1 interactions by delivering PD-L1 siRNA, using various folic acid (FA)-functionalized polyethylenimine (PEI) polymers, to SKOV-3-Luc EOC cells, and investigate the sensitization of the EOC cells to T cell killing. To enhance siRNA uptake into EOC cells, which over express folate receptors, PEI is modified with FA or PEG-FA so that siRNA is complexed into nanoparticles with folate molecules on the surface. PEI modification with a single functional group lowers the polymer cytotoxicity compared to unmodified PEI. FA-conjugated polymers increase siRNA uptake into SKOV-3-luc cells and decrease unspecific uptake into monocytes. All polymers result in 40% to 50% PD-L1 protein knockdown. Importantly, SKOV-3-Luc cells treated with either PEI-FA or PEI- polyethylene glycol (PEG)-FA/PD-L1 siRNA complexes are up to twofold more sensitive to T cell killing compared to scrambled siRNA treated controls. These findings are the first to demonstrate that PD-L1 knockdown in EOC cells, via siRNA/FA-targeted delivery, are able to sensitize cancer cells to T cell killing.

  4. Folate conjugated Mn{sub 3}O{sub 4}@SiO{sub 2} nanoparticles for targeted magnetic resonance imaging in vivo

    SciTech Connect

    Yang, Xinyi; Zhou, Zhiguo; Wang, Li; Tang, Caizhi; Yang, Hong; Yang, Shiping

    2014-09-15

    Graphical abstract: The Mn{sub 3}O{sub 4}@SiO{sub 2}(PEG)–FA has been used as a T{sub 1}-MRI probe for in vivo. - Highlights: • The PEG and FA modified Mn{sub 3}O{sub 4}@SiO{sub 2} nanoparticles (Mn{sub 3}O{sub 4}@SiO{sub 2}–FA) were prepared. • Mn{sub 3}O{sub 4}@SiO{sub 2}–FA exhibited the good colloidal stability in the simulated biological medium. • Mn{sub 3}O{sub 4}@SiO{sub 2}–FA showed the targeting ability to HeLa cells overexpressed the FA receptor. • The T{sub 1}-weighted magnetic resonance (MR) imaging demonstrated the targeting ability of Mn{sub 3}O{sub 4}@SiO{sub 2}–FA in vivo tumor. - Abstract: The monodisperse silica-coated manganese oxide nanoparticles (Mn{sub 3}O{sub 4}@SiO{sub 2} NPs) were synthesized via the high temperature pyrolysis approach and were aminated through silanization. The amine-functionalized Mn{sub 3}O{sub 4} NPs enabled the covalent conjugation of hydrophilic methoxypoly(ethylene glycol) (PEG) and the targeting ligand of folate (FA) onto their surface. The formed PEG and FA modified Mn{sub 3}O{sub 4} NPs (Mn{sub 3}O{sub 4}@SiO{sub 2}(PEG)–FA) exhibited the good colloidal stability in the simulated biological medium and the targeting ability to HeLa cells overexpressed the FA receptor. The T{sub 1}-weighted magnetic resonance (MR) imaging and inductively coupled plasma atomic emission spectroscopy (ICP-AES) analysis of Mn{sub 3}O{sub 4}@SiO{sub 2}(PEG)–FA NPs further demonstrated their targeting ability in tumor.

  5. Biodistribution and pharmacokinetics of Mad2 siRNA-loaded EGFR-targeted chitosan nanoparticles in cisplatin sensitive and resistant lung cancer models

    PubMed Central

    Nascimento, Ana Vanessa; Gattacceca, Florence; Singh, Amit; Bousbaa, Hassan; Ferreira, Domingos; Sarmento, Bruno; Amiji, Mansoor M

    2016-01-01

    Background: The present study focuses on biodistribution profile and pharmacokinetic parameters of EGFR-targeted chitosan nanoparticles (TG CS nanoparticles) for siRNA/cisplatin combination therapy of lung cancer. Material & methods: Mad2 siRNA was encapsulated in EGFR targeted and nontargeted (NTG) CS nanoparticles by electrostatic interaction. The biodistribution of the nanoparticles was assessed qualitatively and quantitatively in cisplatin (DDP) sensitive and resistant lung cancer xenograft model. Results: TG nanoparticles showed a consistent and preferential tumor targeting ability with rapid clearance from the plasma to infiltrate and sustain within the tumor up to 96 h. They exhibit a sixfold higher tumor targeting efficiency compared with the NTG nanoparticles. Conclusion: TG nanoparticles present as an attractive drug delivery platform for RNAi therapeutics against NSCLC. PMID:26980454

  6. Screening target specificity of siRNAs by rapid amplification of cDNA ends (RACE) for non-sequenced species.

    PubMed

    Sabirzhanov, Boris; Sabirzhanova, Inna B; Keifer, Joyce

    2011-05-01

    RNA interference (RNAi) is the process of sequence-specific posttranslational gene silencing triggered by double-stranded RNAs (dsRNAs). RNAi is a widely used approach for studying gene function. However, studies have shown that using siRNA can lead to off-target effects when the siRNA contains sufficient sequence identity to non-target mRNA sequences. One of the important steps in designing dsRNA is verification that it has sequence identity to only the target mRNA. In this report, we propose an approach for primary screening dsRNAs for potential off-target effects by using rapid amplification of cDNA ends. This method can be especially useful for model systems using species that have limited availability of sequence data.

  7. Construction of simple and efficient siRNA validation systems for screening and identification of effective RNAi-targeted sequences from mammalian genes.

    PubMed

    Tsai, Wen-Hui; Chang, Wen-Tsan

    2014-01-01

    RNA interference (RNAi) is an evolutionarily conserved mechanism of gene silencing induced by double-stranded RNAs (dsRNAs). Among the widely used dsRNAs, small interfering RNAs (siRNAs) and short hairpin RNAs have evolved as extremely powerful and the most popular gene silencing reagents. The key challenge to achieving efficient gene silencing especially for the purpose of therapeutics is mainly dependent on the effectiveness and specificity of the selected RNAi-targeted sequences. Practically, only a small number of dsRNAs are capable of inducing highly effective and sequence-specific gene silencing via RNAi mechanism. In addition, the efficiency of gene silencing induced by dsRNAs can only be experimentally examined based on inhibition of the target gene expression. Therefore, it is essential to develop a fully robust and comparative validation system for measuring the efficacy of designed dsRNAs. In this chapter, we focus our discussion on a reliable and quantitative reporter-based siRNA validation system that has been previously established in our laboratory. The system consists of a short synthetic DNA fragment containing an RNAi-targeted sequence of interest and two expression vectors for targeting reporter and triggering siRNA expressions. The efficiency of siRNAs is determined by their abilities to inhibit expression of the targeting reporters with easily quantified readouts including enhanced green fluorescence protein and firefly luciferase. Since only a readily available short synthetic DNA fragment is needed for constructing this reliable and efficient reporter-based siRNA validation system, this system not only provides a powerful strategy for screening highly effective RNAi-targeted sequences from mammalian genes but also implicates the use of RNAi-based dsRNA reagents for reverse functional genomics and molecular therapeutics.

  8. Self-assembled nanoparticles based on the c(RGDfk) peptide for the delivery of siRNA targeting the VEGFR2 gene for tumor therapy

    PubMed Central

    Liu, Li; Liu, Xiaoxia; Xu, Qian; Wu, Ping; Zuo, Xialin; Zhang, Jingjing; Deng, Houliang; Wu, Zhuomin; Ji, Aimin

    2014-01-01

    The clinical application of small interfering RNA (siRNA) has been restricted by their poor intracellular uptake, low serum stability, and inability to target specific cells. During the last several decades, a great deal of effort has been devoted to exploring materials for siRNA delivery. In this study, biodegradable, tumor-targeted, self-assembled peptide nanoparticles consisting of cyclo(Arg–Gly–Asp–d–Phe–Lys)-8–amino–3,6–dioxaoctanoic acid–β–maleimidopropionic acid (hereafter referred to as RPM) were found to be an effective siRNA carrier both in vitro and in vivo. The nanoparticles were characterized based on transmission electron microscopy, circular dichroism spectra, and dynamic light scattering. In vitro analyses showed that the RPM/VEGFR2-siRNA exhibited negligible cytotoxicity and induced effective gene silencing. Delivery of the RPM/VEGFR2 (zebrafish)-siRNA into zebrafish embryos resulted in inhibition of neovascularization. Administration of RPM/VEGFR2 (mouse)-siRNA to tumor-bearing nude mice led to a significant inhibition of tumor growth, a marked reduction of vessels, and a down-regulation of VEGFR2 (messenger RNA and protein) in tumor tissue. Furthermore, the levels of IFN-α, IFN-γ, IL-12, and IL-6 in mouse serum, assayed via enzyme-linked immunosorbent assay, did not indicate any immunogenicity of the RPM/VEGFR2 (mouse)-siRNA in vivo. In conclusion, RPM may provide a safe and effective delivery vector for the clinical application of siRNAs in tumor therapy. PMID:25114522

  9. Fabrication of poly-crystalline Si-based Mie resonators via amorphous Si on SiO2 dewetting

    NASA Astrophysics Data System (ADS)

    Naffouti, Meher; David, Thomas; Benkouider, Abdelmalek; Favre, Luc; Ronda, Antoine; Berbezier, Isabelle; Bidault, Sebastien; Bonod, Nicolas; Abbarchi, Marco

    2016-01-01

    We report the fabrication of Si-based dielectric Mie resonators via a low cost process based on solid-state dewetting of ultra-thin amorphous Si on SiO2. We investigate the dewetting dynamics of a few nanometer sized layers annealed at high temperature to form submicrometric Si-particles. Morphological and structural characterization reveal the polycrystalline nature of the semiconductor matrix as well as rather irregular morphologies of the dewetted islands. Optical dark field imaging and spectroscopy measurements of the single islands reveal pronounced resonant scattering at visible frequencies. The linewidth of the low-order modes can be ~20 nm in full width at half maximum, leading to a quality factor Q exceeding 25. These values reach the state-of-the-art ones obtained for monocrystalline Mie resonators. The simplicity of the dewetting process and its cost-effectiveness opens the route to exploiting it over large scales for applications in silicon-based photonics.

  10. Fabrication of poly-crystalline Si-based Mie resonators via amorphous Si on SiO2 dewetting.

    PubMed

    Naffouti, Meher; David, Thomas; Benkouider, Abdelmalek; Favre, Luc; Ronda, Antoine; Berbezier, Isabelle; Bidault, Sebastien; Bonod, Nicolas; Abbarchi, Marco

    2016-02-07

    We report the fabrication of Si-based dielectric Mie resonators via a low cost process based on solid-state dewetting of ultra-thin amorphous Si on SiO2. We investigate the dewetting dynamics of a few nanometer sized layers annealed at high temperature to form submicrometric Si-particles. Morphological and structural characterization reveal the polycrystalline nature of the semiconductor matrix as well as rather irregular morphologies of the dewetted islands. Optical dark field imaging and spectroscopy measurements of the single islands reveal pronounced resonant scattering at visible frequencies. The linewidth of the low-order modes can be ∼20 nm in full width at half maximum, leading to a quality factor Q exceeding 25. These values reach the state-of-the-art ones obtained for monocrystalline Mie resonators. The simplicity of the dewetting process and its cost-effectiveness opens the route to exploiting it over large scales for applications in silicon-based photonics.

  11. NIR light controlled photorelease of siRNA and its targeted intracellular delivery based on upconversion nanoparticles

    NASA Astrophysics Data System (ADS)

    Yang, Yanmei; Liu, Fang; Liu, Xiaogang; Xing, Bengang

    2012-12-01

    The most notable role of small interfering RNA (siRNA) is in RNA interference (RNAi) and post-transcriptional gene silencing, which leads to a surge of interest in RNAi for both biomedical research and therapeutic applications. However, ``naked'' siRNA cannot cross cellular membranes freely because of highly negative charges which limits its utility for gene therapy. In this work, a system of near-infrared (NIR) light-induced siRNA release from silica coated upconversion nanoparticles (Si-UCNPs) is presented. These Si-UCNPs were functionalized with cationic photocaged linkers through covalent bonding, which could effectively adsorb anionic siRNA through electrostatic attractions and were easily internalized by living cells. Upon NIR light irradiation, the photocaged linker on the Si-UCNPs surface could be cleaved by the upconverted UV light and thus initiated the intracellular release of the siRNA. The in vitro agarose gel electrophoresis and intracellular imaging results indicated that the Si-UCNPs-based gene carrier system allowed effective siRNA delivery and the applications of NIR light instead of direct high energy UV irradiation may greatly guarantee less cell damage.The most notable role of small interfering RNA (siRNA) is in RNA interference (RNAi) and post-transcriptional gene silencing, which leads to a surge of interest in RNAi for both biomedical research and therapeutic applications. However, ``naked'' siRNA cannot cross cellular membranes freely because of highly negative charges which limits its utility for gene therapy. In this work, a system of near-infrared (NIR) light-induced siRNA release from silica coated upconversion nanoparticles (Si-UCNPs) is presented. These Si-UCNPs were functionalized with cationic photocaged linkers through covalent bonding, which could effectively adsorb anionic siRNA through electrostatic attractions and were easily internalized by living cells. Upon NIR light irradiation, the photocaged linker on the Si-UCNPs surface

  12. Solution growth of Si on reorganized porous Si foils and on glass substrates

    NASA Astrophysics Data System (ADS)

    Ehlers, C.; Bansen, R.; Markurt, T.; Uebel, D.; Teubner, Th.; Boeck, T.

    2017-06-01

    We have developed a thin film growth process, which allows for the deposition of closed layers of crystalline Si onto inexpensive substrates in a continuous fashion. Deposition is performed by steady-state solution growth on either reorganized porous Si foils, or on glass substrates with a thin amorphous Si seed layer. The respective monocrystalline and polycrystalline Si films are grown up to a thickness of several ten micrometers, making them suitable for an efficient absorption of sunlight in a photovoltaic device. The structural properties of the Si films have been investigated by scanning electron microscopy (SEM) and transmission electron microscopy (TEM). The determined real structure of the polycrystalline layers with grains in the dimension of several 10 μm offers good prospects to utilize the material as an absorber layer for solar cells.

  13. Efficient and Tumor Targeted siRNA Delivery by Polyethylenimine-graft-polycaprolactone-block-poly(ethylene glycol)-folate (PEI-PCL-PEG-Fol).

    PubMed

    Liu, Li; Zheng, Mengyao; Librizzi, Damiano; Renette, Thomas; Merkel, Olivia M; Kissel, Thomas

    2016-01-04

    Efficient delivery of functional nucleic acids into specific cells or tissues is still a challenge for gene therapy and largely depends on targeted delivery strategies. The folate receptor (FR) is known to be overexpressed extracellularly on a variety of human cancers and is therefore an outstanding gate for tumor-targeted Trojan horse-like delivery of therapeutics. In this study, an amphiphilic and biodegradable ternary copolymer conjugated with folate as ligand, polyethylenimine-graft-polycaprolactone-block-poly(ethylene glycol)-folate (PEI-PCL-PEG-Fol) was synthesized and evaluated for targeted siRNA delivery via folate-FR recognition. The amphiphilic character of similar polymers was shown previously to support endosomal release of endocytosed nanocarriers and to promote formation of long circulating micelles. The obtained PEI-PCL-PEG-Fol exhibited less cytotoxicity in comparison with the corresponding ternary copolymer without folate (PEI-PCL-PEG) and with unmodified PEI25kDa. Stable micelle-like polyplexes with hydrodynamic diameters about 100 nm were found to have a zeta potential of +8.6 mV, which was lower than that of micelleplexes without folate-conjugation (+13-16 mV). Nonetheless, increased cellular uptake and in vitro gene knockdown of PEI-PCL-PEG-Fol/siRNA micelleplexes were observed in SKOV-3 cells, an FR overexpressing cell line, in comparison with the nonfolate-conjugated ones. Moreover, PEI-PCL-PEG-Fol/siRNA micelleplexes exhibited excellent stability in vivo during the analysis of 120 min and a longer circulation half life than hyPEI25kDa/siRNA polyplexes. Most interestingly, the targeted delivery system yielded 17% deposition of the i.v. injected siRNA per gram in the tumor after 24 h due to the effective folate targeting and the prolonged circulation.

  14. Progress on the fabrication of high resolution and lightweight monocrystalline silicon x-ray mirrors

    NASA Astrophysics Data System (ADS)

    Riveros, Raul E.; Biskach, Michael P.; Allgood, Kim D.; Mazzarella, James R.; Sharpe, Marton V.; Zhang, William W.

    2016-07-01

    Monocrystalline silicon is an excellent X-ray mirror substrate material due to its high stiffness, low density, high thermal conductivity, zero internal stress, and commercial availability. Our work at NASA Goddard Space Flight Center focuses on identifying and developing a manufacturing process to produce high resolution and lightweight X-ray mirror segments in a cost and time effective manner. Previous efforts focused on demonstrating the feasibility of cylindrical silicon mirror polishing and lightweighting. Present efforts are aimed towards producing true paraboloidal and hyperboloidal mirror surfaces on the lightweight silicon segments. This paper presents results from these recent investigations, including a mirror which features a surface quality sufficient for a 3 arcsecond telescope.

  15. Numerical analysis of monocrystalline silicon solar cells with fine nanoimprinted textured surface

    NASA Astrophysics Data System (ADS)

    Yoshinaga, Seiya; Ishikawa, Yasuaki; Araki, Shinji; Honda, Tatsuki; Jiang, Yunjiang; Uraoka, Yukiharu

    2017-02-01

    We investigated the surface reflectance of nanoimprinted textures on silicon. Zirconium oxide, which is a wide-bandgap inorganic dielectric material, was used as the texturing material. We performed several calculations to optimize the textures for the production of high-efficiency bulk-type monocrystalline silicon solar cells. Our analysis revealed that nanoimprinted textured solar cells exhibit a lower reverse saturation current density than a solar cell with a conventional etched texture. It was also confirmed that the photocarrier generation rate for a solar cell with a submicron-scale nanoimprinted texture has little dependence on the texture shape. Furthermore, the weighted average reflectance of an optimized nanoimprinted textured solar cell was substantially reduced to 3.72%, suggesting that texture formation by nanoimprint lithography is an extremely effective technology for producing high-efficiency solar cells at a low cost.

  16. Photoconductivities in monocrystalline layered V2O5 nanowires grown by physical vapor deposition

    NASA Astrophysics Data System (ADS)

    Chen, Ruei-San; Wang, Wen-Chun; Chan, Ching-Hsiang; Hsu, Hung-Pin; Tien, Li-Chia; Chen, Yu-Jyun

    2013-10-01

    Photoconductivities of monocrystalline vanadium pentoxide (V2O5) nanowires (NWs) with layered orthorhombic structure grown by physical vapor deposition (PVD) have been investigated from the points of view of device and material. Optimal responsivity and gain for single-NW photodetector are at 7,900 A W-1 and 30,000, respectively. Intrinsic photoconduction (PC) efficiency (i.e., normalized gain) of the PVD-grown V2O5 NWs is two orders of magnitude higher than that of the V2O5 counterpart prepared by hydrothermal approach. In addition, bulk and surface-controlled PC mechanisms have been observed respectively by above- and below-bandgap excitations. The coexistence of hole trapping and oxygen sensitization effects in this layered V2O5 nanostructure is proposed, which is different from conventional metal oxide systems, such as ZnO, SnO2, TiO2, and WO3.

  17. Low cost monocrystalline silicon sheet fabrication for solar cells by advanced ingot technology

    NASA Technical Reports Server (NTRS)

    Fiegl, G. F.; Bonora, A. C.

    1980-01-01

    The continuous liquid feed (CLF) Czochralski furnace and the enhanced I.D. slicing technology for the low-cost production of monocrystalline silicon sheets for solar cells are discussed. The incorporation of the CLF system is shown to improve ingot production rate significantly. As demonstrated in actual runs, higher than average solidification rates (75 to 100 mm/hr for 150 mm 1-0-0 crystals) can be achieved, when the system approaches steady-state conditions. The design characteristics of the CLF furnace are detailed, noting that it is capable of precise control of dopant impurity incorporation in the axial direction of the crystal. The crystal add-on cost is computed to be $11.88/sq m, considering a projected 1986 25-slice per cm conversion factor with an 86% crystal growth yield.

  18. Progress on the Fabrication of High Resolution and Lightweight Monocrystalline Silicon X-ray Mirrors

    NASA Technical Reports Server (NTRS)

    Riveros, Raul E.; Biskach, Michael P.; Allgood, Kim D.; Mazzarella, James R.; Sharpe, Marton V.; Zhang, William W.

    2016-01-01

    Monocrystalline silicon is an excellent X-ray mirror substrate material due to its high stiffness, low density, high thermal conductivity, zero internal stress, and commercial availability. Our work at NASA Goddard Space Flight Center focuses on identifying and developing a manufacturing process to produce high resolution and lightweight X-ray mirror segments in a cost and time effective manner. Previous efforts focused on demonstrating the feasibility of cylindrical silicon mirror polishing and lightweighting. Present efforts are aimed towards producing true paraboloidal and hyperboloidal mirror surfaces on the lightweight silicon segments. This paper presents results from these recent investigations, including a mirror which features a surface quality sufficient for a 3 arcsecond telescope.

  19. Photoconductivities in monocrystalline layered V2O5 nanowires grown by physical vapor deposition

    PubMed Central

    2013-01-01

    Photoconductivities of monocrystalline vanadium pentoxide (V2O5) nanowires (NWs) with layered orthorhombic structure grown by physical vapor deposition (PVD) have been investigated from the points of view of device and material. Optimal responsivity and gain for single-NW photodetector are at 7,900 A W-1 and 30,000, respectively. Intrinsic photoconduction (PC) efficiency (i.e., normalized gain) of the PVD-grown V2O5 NWs is two orders of magnitude higher than that of the V2O5 counterpart prepared by hydrothermal approach. In addition, bulk and surface-controlled PC mechanisms have been observed respectively by above- and below-bandgap excitations. The coexistence of hole trapping and oxygen sensitization effects in this layered V2O5 nanostructure is proposed, which is different from conventional metal oxide systems, such as ZnO, SnO2, TiO2, and WO3. PMID:24160337

  20. Device-based local delivery of siRNA against mammalian target of rapamycin (mTOR) in a murine subcutaneous implant model to inhibit fibrous encapsulation

    PubMed Central

    Takahashi, Hironobu; Wang, Yuwei; Grainger, David W.

    2010-01-01

    Fibrous encapsulation of surgically implant devices is associated with elevated proliferation and activation of fibroblasts in tissues surrounding these implants, frequently causing foreign body complications. Here we test the hypothesis that inhibition of the expression of mammalian target of rapamycin (mTOR) in fibroblasts can mitigate the soft tissue implant foreign body response by suppressing fibrotic responses around implants. In this study, mTOR was knocked down using small interfering RNA conjugated with branched cationic polyethylenimine (bPEI) in fibroblastic lineage cells in serum-based cell culture as shown by both gene and protein analysis. This mTOR knockdown led to an inhibition in fibroblast proliferation by 70% and simultaneous down-regulation in the expression of type I collagen in fibroblasts in vitro. These siRNA/bPEI complexes were released from poly(ethylene glycol) (PEG)-based hydrogel coatings surrounding model polymer implants in a subcutaneous rodent model in vivo. No significant reduction in fibrous capsule thickness and mTOR expression in the foreign body capsules was observed. Observed siRNA inefficacy in this in vivo implant model was attributed to siRNA dosing limitations in the gel delivery system, and lack of targeting ability of the siRNA complex specifically to fibroblasts. While in vitro data supported mTOR knock-down in fibroblast cultures, in vivo siRNA delivery must be further improved to produce clinically relevant effects on fibrotic encapsulation around implants. PMID:20727922

  1. Photomechanical Wave-Driven Delivery of siRNAs Targeting Intermediate Filament Proteins Promotes Functional Recovery after Spinal Cord Injury in Rats

    PubMed Central

    Ando, Takahiro; Sato, Shunichi; Toyooka, Terushige; Kobayashi, Hiroaki; Nawashiro, Hiroshi; Ashida, Hiroshi; Obara, Minoru

    2012-01-01

    The formation of glial scars after spinal cord injury (SCI) is one of the factors inhibiting axonal regeneration. Glial scars are mainly composed of reactive astrocytes overexpressing intermediate filament (IF) proteins such as glial fibrillary acidic protein (GFAP) and vimentin. In the current study, we delivered small interfering RNAs (siRNAs) targeting these IF proteins to SCI model rats using photomechanical waves (PMWs), and examined the restoration of motor function in the rats. PMWs are generated by irradiating a light-absorbing material with 532-nm nanosecond laser pulses from a Q-switched Nd:YAG laser. PMWs can site-selectively increase the permeability of the cell membrane for molecular delivery. Rat spinal cord was injured using a weight-drop device and the siRNA(s) solutions were intrathecally injected into the vicinity of the exposed SCI, to which PMWs were applied. We first confirmed the substantial uptake of fluorescence-labeled siRNA by deep glial cells; then we delivered siRNAs targeting GFAP and vimentin into the lesion. The treatment led to a significant improvement in locomotive function from five days post-injury in rats that underwent PMW-mediated siRNA delivery. This was attributable to the moderate silencing of the IF proteins and the subsequent decrease in the cavity area in the injured spinal tissue. PMID:23272155

  2. Animal models for target diseases in gene therapy--using DNA and siRNA delivery strategies.

    PubMed

    Blagbrough, Ian S; Zara, Chiara

    2009-01-01

    Nanoparticles, including lipopolyamines leading to lipoplexes, liposomes, and polyplexes are targeted drug carrier systems in the current search for a successful delivery system for polynucleic acids. This review is focused on the impact of gene and siRNA delivery for studies of efficacy, pharmacodynamics, and pharmacokinetics within the setting of the wide variety of in vivo animal models now used. This critical appraisal of the recent literature sets out the different models that are currently being investigated to bridge from studies in cell lines through towards clinical reality. Whilst many scientists will be familiar with rodent (murine, fecine, cricetine, and musteline) models, few probably think of fish as a clinically relevant animal model, but zebrafish, madake, and rainbow trout are all being used. Larger animal models include rabbit, cat, dog, and cow. Pig is used both for the prevention of foot-and-mouth disease and human diseases, sheep is a model for corneal transplantation, and the horse naturally develops arthritis. Non-human primate models (macaque, common marmoset, owl monkey) are used for preclinical gene vector safety and efficacy trials to bridge the gap prior to clinical studies. We aim for the safe development of clinically effective delivery systems for DNA and RNAi technologies.

  3. Chitosan-based nanoparticles for survivin targeted siRNA delivery in breast tumor therapy and preventing its metastasis

    PubMed Central

    Sun, Ping; Huang, Wei; Jin, Mingji; Wang, Qiming; Fan, Bo; Kang, Lin; Gao, Zhonggao

    2016-01-01

    Nanoparticle-mediated small interfering RNA (siRNA) delivery is a promising therapeutic strategy in various cancers. However, it is difficult to deliver degradative siRNA to tumor tissue, and thus a safe and efficient vector for siRNA delivery is essential for cancer therapy. In this study, poly(ethylene glycol)-modified chitosan (PEG-CS) was synthesized successfully for delivering nucleic acid drug. We deemed that PEGylated CS could improve its solubility by forming a stable siRNA loaded in nanoparticles, and enhancing transfection efficiency of siRNA-loaded CS nanoparticles in cancer cell line. The research results showed that siRNA loaded in PEGylated CS (PEG-CS/siRNA) nanoparticles with smaller particle size had superior structural stability in the physical environment compared to CS nanoparticles. The data of in vitro antitumor activity revealed that 4T1 tumor cell growth was significantly inhibited and cellular uptake of PEG-CS/siRNA nanoparticles in 4T1 cells was dramatically enhanced compared to naked siRNA groups. The results from flow cytometry and confocal laser scanning microscopy showed that PEG-CS/siRNA nanoparticles were more easily taken up than naked siRNA. Importantly, PEG-CS/siRNA nanoparticles significantly reduced the growth of xenograft tumors of 4T1 cells in vivo. It has been demonstrated that the PEG-CS is a safe and efficient vector for siRNA delivery, and it can effectively reduce tumor growth and prevent metastasis. PMID:27729789

  4. Chitosan-based nanoparticles for survivin targeted siRNA delivery in breast tumor therapy and preventing its metastasis.

    PubMed

    Sun, Ping; Huang, Wei; Jin, Mingji; Wang, Qiming; Fan, Bo; Kang, Lin; Gao, Zhonggao

    Nanoparticle-mediated small interfering RNA (siRNA) delivery is a promising therapeutic strategy in various cancers. However, it is difficult to deliver degradative siRNA to tumor tissue, and thus a safe and efficient vector for siRNA delivery is essential for cancer therapy. In this study, poly(ethylene glycol)-modified chitosan (PEG-CS) was synthesized successfully for delivering nucleic acid drug. We deemed that PEGylated CS could improve its solubility by forming a stable siRNA loaded in nanoparticles, and enhancing transfection efficiency of siRNA-loaded CS nanoparticles in cancer cell line. The research results showed that siRNA loaded in PEGylated CS (PEG-CS/siRNA) nanoparticles with smaller particle size had superior structural stability in the physical environment compared to CS nanoparticles. The data of in vitro antitumor activity revealed that 4T1 tumor cell growth was significantly inhibited and cellular uptake of PEG-CS/siRNA nanoparticles in 4T1 cells was dramatically enhanced compared to naked siRNA groups. The results from flow cytometry and confocal laser scanning microscopy showed that PEG-CS/siRNA nanoparticles were more easily taken up than naked siRNA. Importantly, PEG-CS/siRNA nanoparticles significantly reduced the growth of xenograft tumors of 4T1 cells in vivo. It has been demonstrated that the PEG-CS is a safe and efficient vector for siRNA delivery, and it can effectively reduce tumor growth and prevent metastasis.

  5. Antibody h-R3-dendrimer mediated siRNA has excellent endosomal escape and tumor targeted delivery ability, and represents efficient siPLK1 silencing and inhibition of cell proliferation, migration and invasion

    PubMed Central

    Li, Jun; Liu, Jing; Li, Shengnan; Hao, Yanli; Chen, Lei; Zhang, Xiaoning

    2016-01-01

    The major obstacle to developing siRNA delivery is their extracellular and intracellular barriers. Herein, a humanized anti-EGFR monoclonal antibody h-R3 was developed to modify the self-assembled binary complexes (dendriplexes) of PAMAM and siRNA via electrostatic interactions, and two common ligands HSA and EGF were used as a control. Compared to dendriplexes, h-R3/EGF/HSA-dendriplexes showed increased particle size, decreased zeta potentials and lower cytotoxicity. Moreover, h-R3-dendriplexes presented greater cellular uptake and excellent endosomal escape ability in HepG2 cells. Ex vivo fluorescence imaging revealed that h-R3-dendriplexes showed higher targeted delivery and gene expression in the tumors than dendriplexes, HSA-dendriplexes and EGF-dendriplexes, which was in agreement with confocal results of cryosections. Furthermore, h-R3-dendriplexes for siPLK1 delivery indicated efficient gene silencing, potentiated cell growth inhibition and cell apoptosis, and suppressed cellular migration/invasion. These results indicate that h-R3-dendriplexes represent a great potential to be used as efficient targeted siRNA delivery carriers. PMID:26883109

  6. (Si){sub 5-2y}(AlP){sub y} alloys assembled on Si(100) from Al-P-Si{sub 3} building units

    SciTech Connect

    Watkins, T.; Chizmeshya, A. V. G.; Kouvetakis, J.; Jiang, L.; Xu, C.; Smith, D. J.; Menendez, J.

    2012-01-09

    An original class of IV/III-V hybrid (Si){sub 5-2y}(AlP){sub y}/Si(100) semiconductors have been produced via tailored interactions of molecular P(SiH{sub 3}){sub 3} and atomic Al yielding tetrahedral ''Al-P-Si{sub 3}'' building blocks. Extensive structural, optical, and vibrational characterization corroborates that these units condense to assemble single-phase, monocrystalline alloys containing 60%-90% Si (y = 0.3-1.0) as nearly defect-free layers lattice-matched to Si. Spectroscopic ellipsometry and density functional theory band structure calculations indicate mild compositional bowing of the band gaps, suggesting that the tuning needed for optoelectronic applications should be feasible.

  7. Mg(II)-Catechin nanoparticles delivering siRNA targeting EIF5A2 inhibit bladder cancer cell growth in vitro and in vivo.

    PubMed

    Chen, Zhenhua; Yu, Ting; Zhou, Bangfen; Wei, Jinhuan; Fang, Yong; Lu, Jun; Guo, Ling; Chen, Wei; Liu, Zhi-Ping; Luo, Junhang

    2016-03-01

    Emerging evidence indicates that combination of two or more therapeutic strategies can synergistically enhance antitumor activity in cancer therapy. Here, we established a green method of generating nanocomposite particles that can be fabricated using catechin, a natural anti-cancer compound from green tea, and Mg(2+) in an easy one-step approach at room temperature. We show that Mg(II)-Catechin nanocomposite particles (Mg(II)-Cat NPs) have good biocompatibility and high cellular uptake also can load and effectively deliver small interfering RNA (siRNA) into cells in vitro and to tumor site in vivo. Mg(II)-Cat NPs by themselves had tumor-suppression effects. When complexed with siRNA that targets oncogene eukaryotic translation initiation factor 5A2 (EIF5A2), Mg(II)-Cat/siEIF5A2 complex had further enhanced anti-tumor activity. Mechanistically, we show that Mg(II)-Cat/siEIF5A2 inhibits oncogenic PI3K/Akt signal pathway. More importantly, Mg(II)-Cat/siEIF5A2 had tumor suppression effect in a clinically-relevant rat in-situ bladder cancer model. Our studies demonstrated that combination of Mg(II)-Cat NPs and siRNA is a promising therapeutic modality of combining chemotherapy with gene therapy in order to afford higher therapeutic efficacy and provided a proof of principle for such modality in a pre-clinical setting.

  8. Targeted therapy with MXD3 siRNA, anti-CD22 antibody and nanoparticles for precursor B-cell acute lymphoblastic leukaemia.

    PubMed

    Satake, Noriko; Duong, Connie; Chen, Cathy; Barisone, Gustavo A; Diaz, Elva; Tuscano, Joseph; Rocke, David M; Nolta, Jan; Nitin, Nitin

    2014-11-01

    Conventional chemotherapy for precursor B-cell (preB) acute lymphoblastic leukaemia (ALL) has limitations that could be overcome by targeted therapy. Previously, we discovered a potential therapeutic molecular target, MDX3 (MAX dimerization protein 3), in preB ALL. In this study, we hypothesize that an effective siRNA therapy for preB ALL can be developed using antiCD22 antibody (αCD22 Ab) and nanoparticles. We composed nanocomplexes with super paramagnetic iron oxide nanoparticles (SPIO NPs), αCD22 Abs and MXD3 siRNA molecules based on physical interactions between the molecules. We demonstrated that the MXD3 siRNA-αCD22 Ab-SPIO NP complexes entered leukaemia cells and knocked down MXD3, leading the cells to undergo apoptosis and resulting in decreased live cell counts in the cell line Reh and in primary preB ALL samples in vitro. Furthermore, the cytotoxic effects of the MXD3 siRNA-αCD22 Ab-SPIO NP complexes were significantly enhanced by addition of the chemotherapy drugs vincristine or doxorubicin. We also ruled out potential cytotoxic effects of the MXD3 siRNA-αCD22 Ab-SPIO NP complexes on normal primary haematopoietic cells. Normal B cells were affected while CD34-positive haematopoietic stem cells and non-B cells were not. These data suggest that MXD3 siRNA-αCD22 Ab-SPIO NP complexes have the potential to be a new targeted therapy for preB ALL. © 2014 John Wiley & Sons Ltd.

  9. Fab’-bearing siRNA TNFα-loaded nanoparticles targeted to colonic macrophages offer an effective therapy for experimental colitis

    PubMed Central

    Hamed, Laroui; Emilie, Viennois; Xiao, Bo; Canup, Brandon S.; Duke, Geem; Denning, Timothy L.; Didier, Merlin

    2014-01-01

    Patients suffering from Inflammatory Bowel Disease (IBD) are currently treated by systemic drugs that can have significant side effects. Thus, it would be highly desirable to target TNFα siRNA (a therapeutic molecule) to the inflamed tissue. Here, we demonstrate that TNFα siRNA can be efficiently loaded into nanoparticles (NPs) made of poly (lactic acid) poly (ethylene glycol) block copolymer (PLA-PEG), and that grafting of the Fab’ portion of the F4/80 Ab (Fab’-bearing) onto the NP surface via maleimide/thiol group-mediated covalent bonding improves the macrophage (MP)-targeting kinetics of the NPs to RAW264.7 cells in vitro. Direct binding was shown between MPs and the Fab’-bearing NPs. Next, we orally administered hydrogel (chitosan/alginate)-encapsulated Fab’-bearing TNFα-siRNA-loaded NPs to 3% dextran sodium sulfate (DSS)-treated mice and investigated the therapeutic effect on colitis. In vivo, the release of TNFα-siRNA-loaded NPs into the mouse colon attenuated colitis more efficiently when the NPs were covered with Fab’-bearing, compared to uncovered NPs. All DSS-induced parameters of colonic inflammation (e.g., weight loss, myeloperoxidase activity, and Iκbα accumulation) were more attenuated Fab’-bearing NPs loaded with TNFα siRNA than without the Fab’-bearing. Grafting the Fab’-bearing onto the NPs improved the kinetics of endocytosis as well as the MP-targeting ability, as indicated by flow cytometry. Collectively, our results show that Fab’-bearing PLA-PEG NPs are powerful and efficient nanosized tools for delivering siRNAs into colonic macrophages. PMID:24810114

  10. Immune Activation and Target Organ Damage Are Consequences of Hydrodynamic Treatment but Not Delivery of Naked siRNAs in Mice

    PubMed Central

    Rácz, Zsuzsanna; Godó, Mária; Révész, Csaba

    2011-01-01

    Short-interfering RNAs (siRNAs), key mediators of RNA interference comprise a promising therapeutic tool, although side effects such as interferon (IFN) response are still not perfectly understood. Further, delivery to target organs is a major challenge, possibly associated with side effects including immune activation or organ damage. We investigated whether immune activation as a consequence of double-stranded RNA induced IFN response (Jak/STAT pathway activation or cytokine production) or target organ damage is induced by in vivo low-volume (LV) or high-volume (HV) hydrodynamic delivery or treatment with naked siRNA. NMRI mice were injected with naked siRNAs or saline by hydrodynamic injection (HDI) and positive control mice received polyinosinic–polycytidilic acid (poly I:C). LV (1 mL/mouse) and HV (10% of body weight) HDI were compared. After LV HDI, STAT1 and OAS1 gene expression inflammatory cytokine plasma levels and target organ injury were assessed. LV HDI induced slight alanine aminotransferase elevation and mild hepatocyte injury, whereas HV HDI resulted in high ALAT level and extensive hepatocyte necrosis. STAT1 or OAS1 was not induced by LV siRNA; however, HV saline led to a time-dependent slight increase in gene expression. Inflammatory cytokine plasma level and organ histology and functional parameters demonstrated no damage following LV HDI with or without siRNA. Our data demonstrate that naked siRNAs may be harnessed, without the induction of IFN response or immune activation, and that LV HDI is preferable, because HV HDI may cause organ damage. PMID:21749298

  11. Targeted therapy with MXD3 siRNA, anti-CD22 antibody and nanoparticles for precursor B-cell acute lymphoblastic leukaemia

    PubMed Central

    Satake, Noriko; Duong, Connie; Chen, Cathy; Barisone, Gustavo A.; Diaz, Elva; Tuscano, Joseph; Rocke, David M.; Nolta, Jan; Nitin, Nitin

    2014-01-01

    Summary Conventional chemotherapy for precursor B-cell (preB) acute lymphoblastic leukaemia (ALL) has limitations that could be overcome by targeted therapy. Previously, we discovered a potential therapeutic molecular target, MDX3 (MAX dimerization protein 3), in preB ALL. In this study, we hypothesize that an effective siRNA therapy for preB ALL can be developed using antiCD22 antibody (αCD22 Ab) and nanoparticles. We composed nanocomplexes with super paramagnetic iron oxide nanoparticles (SPIO NPs), αCD22 Abs and MXD3 siRNA molecules based on physical interactions between the molecules. We demonstrated that the MXD3 siRNA-αCD22 Ab-SPIO NP complexes entered leukaemia cells and knocked down MXD3, leading the cells to undergo apoptosis and resulting in decreased live cell counts in the cell line Reh and in primary preB ALL samples in vitro. Furthermore, the cytotoxic effects of the MXD3 siRNA-αCD22 Ab-SPIO NP complexes were significantly enhanced by addition of the chemotherapy drugs vincristine or doxorubicin. We also ruled out potential cytotoxic effects of the MXD3 siRNA-αCD22 Ab-SPIO NP complexes on normal primary haematopoietic cells. Normal B cells were affected while CD34-positive haematopoietic stem cells and non-B cells were not. These data suggest that MXD3 siRNA-αCD22 Ab-SPIO NP complexes have the potential to be a new targeted therapy for preB ALL. PMID:25196579

  12. Fab'-bearing siRNA TNFα-loaded nanoparticles targeted to colonic macrophages offer an effective therapy for experimental colitis.

    PubMed

    Laroui, Hamed; Viennois, Emilie; Xiao, Bo; Canup, Brandon S B; Geem, Duke; Denning, Timothy L; Merlin, Didier

    2014-07-28

    Patients suffering from inflammatory bowel disease (IBD) are currently treated by systemic drugs that can have significant side effects. Thus, it would be highly desirable to target TNFα siRNA (a therapeutic molecule) to the inflamed tissue. Here, we demonstrate that TNFα siRNA can be efficiently loaded into nanoparticles (NPs) made of poly (lactic acid) poly (ethylene glycol) block copolymer (PLA-PEG), and that grafting of the Fab' portion of the F4/80 Ab (Fab'-bearing) onto the NP surface via maleimide/thiol group-mediated covalent bonding improves the macrophage (MP)-targeting kinetics of the NPs to RAW264.7 cells in vitro. Direct binding was shown between MPs and the Fab'-bearing NPs. Next, we orally administered hydrogel (chitosan/alginate)-encapsulated Fab'-bearing TNFα-siRNA-loaded NPs to 3% dextran sodium sulfate (DSS)-treated mice and investigated the therapeutic effect on colitis. In vivo, the release of TNFα-siRNA-loaded NPs into the mouse colon attenuated colitis more efficiently when the NPs were covered with Fab'-bearing, compared to uncovered NPs. All DSS-induced parameters of colonic inflammation (e.g., weight loss, myeloperoxidase activity, and Iκbα accumulation) were more attenuated Fab'-bearing NPs loaded with TNFα siRNA than without the Fab'-bearing. Grafting the Fab'-bearing onto the NPs improved the kinetics of endocytosis as well as the MP-targeting ability, as indicated by flow cytometry. Collectively, our results show that Fab'-bearing PLA-PEG NPs are powerful and efficient nanosized tools for delivering siRNAs into colonic macrophages.

  13. A Poly(Propyleneimine) Dendrimer-Based Polyplex-System for Single-Chain Antibody-Mediated Targeted Delivery and Cellular Uptake of SiRNA.

    PubMed

    Tietze, Stefanie; Schau, Isabell; Michen, Susanne; Ennen, Franka; Janke, Andreas; Schackert, Gabriele; Aigner, Achim; Appelhans, Dietmar; Temme, Achim

    2017-07-01

    Therapeutics based on small interfering RNAs (siRNAs) offer a great potential to treat so far incurable diseases or metastatic cancer. However, the broad application of siRNAs using various nonviral carrier systems is hampered by unspecific toxic side effects, poor pharmacokinetics due to unwanted delivery of siRNA-loaded nanoparticles into nontarget organs, or rapid renal excretion. In order to overcome these obstacles, several targeting strategies using chemically linked antibodies and ligands have emerged. This study reports a new modular polyplex carrier system for targeted delivery of siRNA, which is based on transfection-disabled maltose-modified poly(propyleneimine)-dendrimers (mal-PPI) bioconjugated to single chain fragment variables (scFvs). To achieve targeted delivery into tumor cells expressing the epidermal growth factor receptor variant III (EGFRvIII), monobiotinylated anti-EGFRvIII scFv fused to a Propionibacterium shermanii transcarboxylase-derived biotinylation acceptor (P-BAP) is bioconjugated to mal-PPI through a novel coupling strategy solely based on biotin-neutravidin bridging. In contrast to polyplexes containing an unspecific control scFv-P-BAP, the generated EGFRvIII-specific polyplexes are able to exclusively deliver siRNA to tumor cells and tumors by receptor-mediated endocytosis. These results suggest that receptor-mediated uptake of otherwise noninternalized mal-PPI-based polyplexes is a promising avenue to improve siRNA therapy of cancer, and introduce a novel strategy for modular bioconjugation of protein ligands to nanoparticles. © 2017 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  14. CONDENSED MATTER: STRUCTURE, THERMAL AND MECHANICAL PROPERTIES: SiC based Si/SiC heterojunction and its rectifying characteristics

    NASA Astrophysics Data System (ADS)

    Zhu, Feng; Chen, Zhi-Ming; Li, Lian-Bi; Zhao, Shun-Feng; Lin, Tao

    2009-11-01

    The Si on SiC heterojunction is still poorly understood, although it has a number of potential applications in electronic and optoelectronic devices, for example, light-activated SiC power switches where Si may play the role of an light absorbing layer. This paper reports on Si films heteroepitaxially grown on the Si face of (0001) n-type 6H-SiC substrates and the use of B2H6 as a dopant for p-Si grown at temperatures in a range of 700-950 °C. X-ray diffraction (XRD) analysis and transmission electron microscopy (TEM) tests have demonstrated that the samples prepared at the temperatures ranged from 850 °C to 900 °C are characterized as monocrystalline silicon. The rocking XRD curves show a well symmetry with FWHM of 0.4339° Omega. Twin crystals and stacking faults observed in the epitaxial layers might be responsible for widening of the rocking curves. Dependence of the crystal structure and surface topography on growth temperature is discussed based on the experimental results. The energy band structure and rectifying characteristics of the Si/SiC heterojunctions are also preliminarily tested.

  15. In vivo efficacy and off-target effects of locked nucleic acid (LNA) and unlocked nucleic acid (UNA) modified siRNA and small internally segmented interfering RNA (sisiRNA) in mice bearing human tumor xenografts.

    PubMed

    Mook, Orf; Vreijling, Jeroen; Wengel, Suzy L; Wengel, Jesper; Zhou, Chuanzheng; Chattopadhyaya, Jyoti; Baas, Frank; Fluiter, Kees

    2010-07-01

    The clinical use of small interfering RNA (siRNA) is hampered by poor uptake by tissues and instability in circulation. In addition, off-target effects pose a significant additional problem for therapeutic use of siRNA. Chemical modifications of siRNA have been reported to increase stability and reduce off-target effects enabling possible therapeutic use of siRNA. Recently a large scale direct comparison of the impact of 21 different types of novel chemical modifications on siRNA efficiency and cell viability was published.1 It was found that several types of chemical modifications could enhance siRNA activity beyond that of an unmodified siRNA in vitro. In addition, a novel siRNA design, termed small internally segmented interfering RNA (sisiRNA), composed of an intact antisense strand and segmented guide strand stabilized using LNA was shown to be effective in cell based assays. In the present study we examined the in vivo efficacy of the LNA and UNA modified siRNA and sisiRNA in a mouse model bearing human tumor xenografts. We studied the biodistribution and efficacy of target knockdown in the mouse model. In addition we used whole genome profiling to assess the off-target effects in the liver of the mouse and the tumor xenografts. We report that LNA and UNA modified siRNA and sisiRNA improve the efficacy in target knockdown as compared with unmodified siRNA in the tumor xenografts without formulation. However, the level of off-target gene regulation in both the tumor and the liver correlated with the increase in efficacy in target knockdown, unless the seed region of the siRNA was modified.

  16. Synthetic siRNA targeting the breakpoint of EWS/Fli-1 inhibits growth of Ewing sarcoma xenografts in a mouse model.

    PubMed

    Takigami, Iori; Ohno, Takatoshi; Kitade, Yukio; Hara, Akira; Nagano, Akihito; Kawai, Gou; Saitou, Mitsuru; Matsuhashi, Aya; Yamada, Kazunari; Shimizu, Katsuji

    2011-01-01

    The EWS/Fli-1 fusion gene, a product of the translocation t(11;22, q24;q12), is detected in 85% of Ewing sarcomas and primitive neuroectodermal tumors. It is thought to be a transcriptional activator that plays a significant role in tumorigenesis. In this study, we developed a novel EWS/Fli-1 blockade system using RNA interference and tested its application for inhibiting the proliferation of Ewing sarcoma cells in vitro and the treatment of mouse tumor xenografts in vivo. We designed and synthesized a small interfering RNA (siRNA) possessing an aromatic compound at the 3'-end targeting the breakpoint of EWS/Fli-1. As this sequence is present only in tumor cells, it is a potentially relevant target. We found that the siRNA targeting EWS/Fli-1 significantly suppressed the expression of EWS/Fli-1 protein sequence specifically and also reduced the expression of c-Myc protein in Ewing sarcoma cells. We further demonstrated that inhibition of EWS/Fli-1 expression efficiently inhibited the proliferation of the transfected cells but did not induce apoptotic cell death. In addition, the siRNA possessing the aromatic compound at the 3'-end was more resistant to nucleolytic degradation than the unmodified siRNA. Administration of the siRNA with atelocollagen significantly inhibited the tumor growth of TC-135, a Ewing sarcoma cell line, which had been subcutaneously xenografted into mice. Moreover, modification of the 3'-end with an aromatic compound improved its efficiency in vivo. Our data suggest that specific downregulation of EWS/Fli-1 by RNA interference is a possible approach for the treatment of Ewing sarcoma.

  17. Folic acid-conjugated GdPO4:Tb3+@SiO2 Nanoprobe for folate receptor-targeted optical and magnetic resonance bi-modal imaging

    NASA Astrophysics Data System (ADS)

    Xu, Xianzhu; Zhang, Xiaoying; Wu, Yanli

    2016-11-01

    Both fluorescent and magnetic nanoprobes have great potential applications for diagnostics and therapy. In the present work, a folic acid-conjugated and silica-modified GdPO4:Tb3+ (GdPO4:Tb3+@SiO2-FA) dual nanoprobe was strategically designed and synthesized for the targeted dual-modality optical and magnetic resonance (MR) imaging via a facile aqueous method. Their structural, optical, and magnetic properties were determined using transmission electron microscopy (TEM), X-ray diffraction (XRD), Fourier transform infrared (FTIR), ultraviolet-visible spectra (UV-Vis), photoluminescence (PL), and superconducting quantum interference device (SQUID). These results indicated that GdPO4:Tb3+@SiO2-FA were uniform monodisperse core-shell structured nanorods (NRs) with an average length of 200 nm and an average width of 25 nm. The paramagnetic property of the synthesized GdPO4:Tb3+@SiO2-FA NRs was confirmed with its linear hysteresis plot (M-H). In addition, the NRs displayed an obvious T1-weighted effect and thus it could potentially serve as a T1-positive contrast agent. The NRs emitted green lights due to the 5D4 → 7F5 transition of the Tb3+. The in vitro assays with NCI-H460 lung cancer cells and human embryonic kidney cell line 293T cells indicated that the GdPO4:Tb3+@SiO2-FA nanoprobe could specifically bind the cells bearing folate receptors (FR). The MTT assay of the NRs revealed that its cytotoxicity was very low. Further in vivo MRI experiments distinctively depict enhanced anatomical features in a xenograft tumor. These results suggest that the GdPO4:Tb3+@SiO2-FA NPs have excellent imaging and cell-targeting abilities for the folate receptor-targeted dual-modality optical and MR imaging and can be potentially used as the nanoprobe for bioimaging.

  18. Focused ion beam supported growth of monocrystalline wurtzite InAs nanowires grown by molecular beam epitaxy

    NASA Astrophysics Data System (ADS)

    Scholz, Sven; Schott, Rüdiger; Labud, Patrick A.; Somsen, Christoph; Reuter, Dirk; Ludwig, Arne; Wieck, Andreas D.

    2017-07-01

    We investigate monocrystalline InAs nanowires (NWs) which are grown catalyst assisted by molecular beam epitaxy (MBE) and create the catalyst by focused ion beam (FIB) implanted Au spots. With this combination of methods an aspect ratio, i.e. the length to width ratio, of the grown NWs up to 300 was achieved. To control the morphology and crystalline structure of the NWs, the growth parameters like temperature, flux ratios and implantation fluence are varied and optimized. Furthermore, the influence of the used molecular arsenic species, in particular the As2 to As4 ratio, is investigated and adjusted. In addition to the high aspect ratio, this optimization results in the growth of monocrystalline InAs NWs with a negligible number of stacking faults. Single NWs were placed site-controlled by FIB implantation, which supplements the working field of area growth.

  19. Systemic siRNA Delivery via Peptide-Tagged Polymeric Nanoparticles, Targeting PLK1 Gene in a Mouse Xenograft Model of Colorectal Cancer

    PubMed Central

    Malhotra, Meenakshi; Tomaro-Duchesneau, Catherine; Saha, Shyamali; Prakash, Satya

    2013-01-01

    Polymeric nanoparticles were developed from a series of chemical reactions using chitosan, polyethylene glycol, and a cell-targeting peptide (CP15). The nanoparticles were complexed with PLK1-siRNA. The optimal siRNA loading was achieved at an N : P ratio of 129.2 yielding a nanoparticle size of >200 nm. These nanoparticles were delivered intraperitoneally and tested for efficient delivery, cytotoxicity, and biodistribution in a mouse xenograft model of colorectal cancer. Both unmodified and modified chitosan nanoparticles showed enhanced accumulation at the tumor site. However, the modified chitosan nanoparticles showed considerably, less distribution in other organs. The relative gene expression as evaluated showed efficient delivery of PLK1-siRNA (0.5 mg/kg) with 50.7 ± 19.5% knockdown (P = 0.031) of PLK1 gene. The in vivo data reveals no systemic toxicity in the animals, when tested for systemic inflammation and liver toxicity. These results indicate a potential of using peptide-tagged nanoparticles for systemic delivery of siRNA at the targeted tumor site. PMID:24159333

  20. Co-delivery of doxorubicin and siRNA for glioma therapy by a brain targeting system: angiopep-2-modified poly(lactic-co-glycolic acid) nanoparticles.

    PubMed

    Wang, Lei; Hao, Yongwei; Li, Haixia; Zhao, Yalin; Meng, Dehui; Li, Dong; Shi, Jinjin; Zhang, Hongling; Zhang, Zhenzhong; Zhang, Yun

    2015-01-01

    It is very challenging to treat brain cancer because of the blood-brain barrier (BBB) restricting therapeutic drug or gene to access the brain. In this research project, angiopep-2 (ANG) was used as a brain-targeted peptide for preparing multifunctional ANG-modified poly(lactic-co-glycolic acid) (PLGA) nanoparticles (NPs), which encapsulated both doxorubicin (DOX) and epidermal growth factor receptor (EGFR) siRNA, designated as ANG/PLGA/DOX/siRNA. This system could efficiently deliver DOX and siRNA into U87MG cells leading to significant cell inhibition, apoptosis and EGFR silencing in vitro. It demonstrated that this drug system was capable of penetrating the BBB in vivo, resulting in more drugs accumulation in the brain. The animal study using the brain orthotopic U87MG glioma xenograft model indicated that the ANG-targeted co-delivery of DOX and EGFR siRNA resulted in not only the prolongation of the life span of the glioma-bearing mice but also an obvious cell apoptosis in glioma tissue.

  1. siRNAs targeting PB2 and NP genes potentially inhibit replication of Highly Pathogenic H5N1 Avian Influenza Virus.

    PubMed

    Behera, Padmanava; Nagarajan, Shanmugasundaram; Murugkar, Harshad V; Kalaiyarasu, Semmannan; Prakash, Anil; Gothalwal, Ragini; Dubey, Shiv Chandra; Kulkarni, Diwakar D; Tosh, Chakradhar

    2015-06-01

    Highly Pathogenic Avian Influenza (HPAI) H5N1 virus is a threat to animal and public health worldwide. Till date, the H5N1 virus has claimed 402 human lives, with a mortality rate of 58 percent and has caused the death or culling of millions of poultry since 2003. In this study, we have designed three siRNAs (PB2-2235, PB2-479 and NP-865) targeting PB2 and NP genes of avian influenza virus and evaluated their potential, measured by hemagglutination (HA), plaque reduction and Real time RT-PCR assay, in inhibiting H5N1 virus (A/chicken/Navapur/7972/2006) replication in MDCK cells. The siRNAs caused 8- to 16-fold reduction in virus HA titers at 24 h after challenged with 100TCID50 of virus. Among these siRNAs, PB2-2235 offered the highest inhibition of virus replication with 16-fold reduction in virus HA titer, 80 percent reduction in viral plaque counts and 94 percent inhibition in expression of specific RNA at 24 h. The other two siRNAs had 68-73 percent and 87-88 percent reduction in viral plaque counts and RNA copy number, respectively. The effect of siRNA on H5N1 virus replication continued till 48h (maximum observation period). These findings suggest that PB2-2235 could efficiently inhibit HPAI H5N1 virus replication.

  2. Targeted partial surface modification with nano-SiO2@Li2CoPO4F as high-voltage cathode material for LIBs

    NASA Astrophysics Data System (ADS)

    Chang, Caiyun; Huang, Zhipeng; Tian, Runsai; Jiang, Xinyu; Li, Chunsheng; Feng, Jijun

    2017-10-01

    Tuning whole/partial surface modification on cathode material with oxide material is a sought-after method to enhance the electrochemical performance in power storage field. Herein, nano-SiO2 targeted partial surface modified high voltage cathode material Li2CoPO4F has been successfully fabricated via a facile self-assembly process in silica dispersion at ambient temperature. With the aid of polar -OH groups attracted on the surface of SiO2 micelles, the nano-SiO2 preferentially nestle up along the borders and boundaries of Li2CoPO4F particles, where protection should be deployed with emphasis against the undesirable interactions between materials and electrolytes. Compared with pristine Li2CoPO4F, the SiO2 selectively modified Li2CoPO4F cathode materials, especially LCPF-3S, exhibit desirable electrochemical performances with higher discharge capacity, more outstanding cycle stability and favorable rate capability without any additional carbon involved. The greatly enhanced electrochemical properties can be attributed to the improved lithium-ion diffusion kinetics and structure tolerance during repeated lithiation/delithiation process. Such findings reveal a great potential of nano-SiO2 modified Li2CoPO4F as high energy cathode material for lithium ion batteries.

  3. Study of Rayleigh–Taylor growth in laser irradiated planar SiO{sub 2} targets at ignition-relevant conditions

    SciTech Connect

    Hager, J. D.; Collins, T. J. B.; Knauer, J. P.; Meyerhofer, D. D.; Sangster, T. C.; Smalyuk, V. A.

    2013-07-15

    Rayleigh–Taylor (RT) growth experiments were performed on the OMEGA laser [T. R. Boehly et al., Opt. Commun. 133, 495 (1997)] using planar SiO{sub 2} targets seeded with a single mode 60-μm wavelength perturbation driven at peak laser intensities up to 9 × 10{sup 14} W/cm{sup 2}. These are the first RT measurements in SiO{sub 2} at conditions relevant to direct-drive inertial confinement fusion ignition. The measured average modulation growth rates agree with the 2-D hydrodynamics code DRACO, providing an important step in the development of target ablators that are robust to RT growth and hot- electron preheat considerations when driven at the intensities required to achieve thermonuclear ignition.

  4. pH-labile PEGylation of siRNA-loaded lipid nanoparticle improves active targeting and gene silencing activity in hepatocytes.

    PubMed

    Hashiba, Kazuki; Sato, Yusuke; Harashima, Hideyoshi

    2017-09-28

    Lipid nanoparticles (LNPs) are one of the promising technologies for the in vivo delivery of short interfering RNA (siRNA). Modifying LNPs with polyethyleneglycol (PEG) is widely used to inhibit non-specific interactions with serum components in the blood stream, and is a useful strategy for maximizing the efficiency of active targeting. However, it is a widely accepted fact that PEGylation of the LNP surface strongly inhibits fusion between LNPs and endosomal membranes, resulting in poor cytosolic siRNA delivery, a process that is referred to as the 'PEG-dilemma'. In the present study, in an attempt to overcome this problem, siRNA-loaded LNPs were modified with PEG through maleic anhydride, a pH-labile linkage. The in vitro, suppression of cationic charge, stealth function at physiological pH up to 1h and the rapid desorption of PEG and restoration of fusogenic activity under slightly acidic conditions (within only 2min) were achieved by PEG modification of the LNPs through maleic anhydride. In vivo, PEG modification through maleic anhydride resulted in a dramatic improvement in the targeting capability of the active targeting of ligand (N-acetyl-d-galactosamine)-modified LNPs to hepatocytes, with an approximately 14-fold increase in gene silencing activity in factor 7 model mice. Taken together, the maleic anhydride-mediated pH-labile PEGylation of the active targeting LNPs is a useful strategy for achieving the specific and efficient delivery of siRNAs in vivo. Copyright © 2017 Elsevier B.V. All rights reserved.

  5. Targeted systemic delivery of siRNA to cervical cancer model using cyclic RGD-installed unimer polyion complex-assembled gold nanoparticles.

    PubMed

    Yi, Yu; Kim, Hyun Jin; Mi, Peng; Zheng, Meng; Takemoto, Hiroyasu; Toh, Kazuko; Kim, Beob Soo; Hayashi, Kotaro; Naito, Mitsuru; Matsumoto, Yu; Miyata, Kanjiro; Kataoka, Kazunori

    2016-12-28

    For systemic delivery of small interfering RNA (siRNA) to solid tumors, we developed an actively-targeted unimer polyion complex-assembled gold nanoparticle (uPIC-AuNP) by a two-step assembling process. First is the monodispersed uPIC formation from the single molecules of therapeutic siRNA and the block catiomer, cyclic RGD (cRGD) peptide-installed poly(ethylene glycol)-block-poly(l-lysine) modified with lipoic acid (LA) at the ω-end (cRGD-PEG-PLL-LA). Second is the surface decoration of a 20nm-sized AuNP with uPICs. The cRGD-installed uPIC-AuNPs (cRGD-uPIC-AuNP) provided the targetability for selective binding to the cancer and cancer-related endothelial cellular surface, while regulating their size <50nm with a quite narrow distribution. The targeting efficacy of the cRGD-uPIC-AuNP was confirmed by in vitro cellular uptake in cultured cervical cancer (HeLa) cells and in vivo tumor accumulation in a subcutaneous HeLa model after systemic administration, compared with a non-targeted control uPIC-AuNP. Due to the targetability of the ligand, the cRGD-uPIC-AuNP achieved the significantly enhanced gene silencing ability in the subcutaneous HeLa tumor. Ultimately, the systemic delivery of siRNA targeted for papilloma virus-derived E6 oncogene by cRGD-uPIC-AuNP significantly inhibited the growth of subcutaneous HeLa tumor. This research demonstrates that the bottom-up construction of nanocarriers using monodispersed building blocks can be employed as delivery platforms for RNA interference-based cancer therapy.

  6. Control of cancer stem cell like population by intracellular target identification followed by the treatment with peptide-siRNA complex.

    PubMed

    Suh, Jin Sook; Lee, Hyun Jung; Nam, Hyun; Jo, Beom Soo; Lee, Dong Woo; Kim, Ji-Hye; Lee, Jue Yeon; Chung, Chong Pyoung; Lee, Gene; Park, Yoon Jeong

    2017-09-23

    Cancer stem cells (CSCs) are a subpopulation of cancer cells and have been known to create cancer reoccurrence during cancer therapy due to their stem cell-like characteristics. However, exact target to control the CSC has not been fully established. Here, we enriched CD44(High) population of MDA-MB-231 cells by CD44 antibody as a CSC marker. By Phospho Antibody Array, CD44(High) population of MDA-MB-231 cells reveals Feline sarcoma-related tyrosine kinase (FER) protein was highly activated. When FER siRNA and low molecular weight protamine (LMWP) as cell penetrating peptides are applied to this population, cancer migration and colony forming ability are inhibited. Moreover, silencing FER using FER siRNA and LMWP conjugates enhances anti-metastasis related factors including E-cadherin, p75 and p63. Taken together, FER is a new marker for targeting breast CSCs and peptide-mediated siRNA method could be an effective and safe way of delivery and be a new therapeutic strategy for targeting breast cancer. Copyright © 2017. Published by Elsevier Inc.

  7. VRP09 Reduction of Corneal Scarring Following Blast and Burn Injuries to Cornea Using siRNAs Targeting TGFb and CTGF

    DTIC Science & Technology

    2013-03-01

    aSMA ) synthesis. Second, we proposed to develop an advanced ex vivo organ culture system using viable explants of rabbit corneas, and assess the...effect of the most effective triple siRNA combination for reduction of target genes, collagen and alpha smooth muscle actin ( aSMA ) in rabbit corneas...targeting three key genes (TGFb, TGFbRII, and CTGF) that synergistically reduces the level of mRNAs for type I collagen gene and aSMA by >95% without

  8. ICAM-1-Targeted, Lcn2 siRNA-Encapsulating Liposomes are Potent Anti-angiogenic Agents for Triple Negative Breast Cancer

    PubMed Central

    Guo, Peng; Yang, Jiang; Jia, Di; Moses, Marsha A.; Auguste, Debra T.

    2016-01-01

    Lipocalin 2 (Lcn2) is a promising therapeutic target as well as a potential diagnostic biomarker for breast cancer. It has been previously shown to promote breast cancer progression by inducing the epithelial to mesenchymal transition in breast cancer cells as well as by enhancing angiogenesis. Lcn2 levels in urine and tissue samples of breast cancer patients has also been correlated with breast cancer status and poor patient prognosis. In this study, we have engineered a novel liposomal small interfering RNA (siRNA) delivery system to target triple negative breast cancer (TNBC) via a recently identified molecular target, intercellular adhesion molecule-1 (ICAM-1). This ICAM-1-targeted, Lcn2 siRNA- encapsulating liposome (ICAM-Lcn2-LP) binds human TNBC MDA-MB-231cells significantly stronger than non-neoplastic MCF-10A cells. Efficient Lcn2 knockdown by ICAM-Lcn2-LPs led to a significant reduction in the production of vascular endothelial growth factor (VEGF) from MDA-MB-231 cells, which, in turn, led to reduced angiogenesis both in vitro and in vivo. Angiogenesis (neovascularization) is a requirement for solid tumor growth and progression, and its inhibition is an important therapeutic strategy for human cancers. Our results indicate that a tumor-specific strategy such as the TNBC-targeted, anti-angiogenic therapeutic approach developed here, may be clinically useful in inhibiting TNBC progression. PMID:26722369

  9. Biocompatible and colloidally stabilized mPEG-PE/calcium phosphate hybrid nanoparticles loaded with siRNAs targeting tumors

    PubMed Central

    Gao, Pei; Zhang, Xiangyu; Wang, Hongzhi; Zhang, Qinghong

    2016-01-01

    Calcium phosphate nanoparticles are safe and effective delivery vehicles for small interfering RNA (siRNA), as a result of their excellent biocompatibility. In this work, mPEG-PE (polyethylene glycol-L-α-phosphatidylethanolamine) was synthesized and used to prepare nanoparticles composed of mPEG-PE and calcium phosphate for siRNA delivery. Calcium phosphate and mPEG-PE formed the stable hybrid nanoparticles through self-assembly resulting from electrostatic interaction in water. The average size of the hybrid nanoparticles was approximately 53.2 nm with a negative charge of approximately −16.7 mV, which was confirmed by dynamic light scattering (DLS) measurements. The nanoparticles exhibited excellent stability in serum and could protect siRNA from ribonuclease (RNase) degradation. The cellular internalization of siRNA-loaded nanoparticles was evaluated in SMMC-7721 cells using a laser scanning confocal microscope (CLSM) and flow cytometry. The hybrid nanoparticles could efficiently deliver siRNA to cells compared with free siRNA. Moreover, the in vivo distribution of Cy5-siRNA-loaded hybrid nanoparticles was observed after being injected into tumor-bearing nude mice. The nanoparticles concentrated in the tumor regions through an enhanced permeability and retention (EPR) effect based on the fluorescence intensities of tissue distribution. A safety evaluation of the nanoparticles was performed both in vitro and in vivo demonstrating that the hybrid nanoparticle delivery system had almost no toxicity. These results indicated that the mPEG-PE/CaP hybrid nanoparticles could be a stable, safe and promising siRNA nanocarrier for anticancer therapy. PMID:26625203

  10. Biocompatible and colloidally stabilized mPEG-PE/calcium phosphate hybrid nanoparticles loaded with siRNAs targeting tumors.

    PubMed

    Gao, Pei; Zhang, Xiangyu; Wang, Hongzhi; Zhang, Qinghong; Li, He; Li, Yaogang; Duan, Yourong

    2016-01-19

    Calcium phosphate nanoparticles are safe and effective delivery vehicles for small interfering RNA (siRNA), as a result of their excellent biocompatibility. In this work, mPEG-PE (polyethylene glycol-L-α-phosphatidylethanolamine) was synthesized and used to prepare nanoparticles composed of mPEG-PE and calcium phosphate for siRNA delivery. Calcium phosphate and mPEG-PE formed the stable hybrid nanoparticles through self-assembly resulting from electrostatic interaction in water. The average size of the hybrid nanoparticles was approximately 53.2 nm with a negative charge of approximately -16.7 mV, which was confirmed by dynamic light scattering (DLS) measurements. The nanoparticles exhibited excellent stability in serum and could protect siRNA from ribonuclease (RNase) degradation. The cellular internalization of siRNA-loaded nanoparticles was evaluated in SMMC-7721 cells using a laser scanning confocal microscope (CLSM) and flow cytometry. The hybrid nanoparticles could efficiently deliver siRNA to cells compared with free siRNA. Moreover, the in vivo distribution of Cy5-siRNA-loaded hybrid nanoparticles was observed after being injected into tumor-bearing nude mice. The nanoparticles concentrated in the tumor regions through an enhanced permeability and retention (EPR) effect based on the fluorescence intensities of tissue distribution. A safety evaluation of the nanoparticles was performed both in vitro and in vivo demonstrating that the hybrid nanoparticle delivery system had almost no toxicity. These results indicated that the mPEG-PE/CaP hybrid nanoparticles could be a stable, safe and promising siRNA nanocarrier for anticancer therapy.

  11. Lentivirus vectors construction of SiRNA targeting interference GPC3 gene and its biological effects on liver cancer cell lines Huh-7.

    PubMed

    Lei, Chang-Jiang; Yao, Chun; Pan, Qing-Yun; Long, Hao-Cheng; Li, Lei; Zheng, Shu-Ping; Zeng, Cheng; Huang, Jian-Bin

    2014-10-01

    To build GPC3 gene short hairpin interference RNA (shRNA) slow virus vector, observe expression of Huh-7 GPC3 gene in human liver cell line proliferation apoptosis and the effect of GPC3 gene influencing on liver cancer cell growth, and provide theoretical basis for gene therapy of liver cancer. Hepatocellular carcinoma cell line Huh-7 was transfected by a RNA interference technique. GPC3 gene expression in a variety of liver cancer cell lines was detected by fluorescence quantitative PCR. Targeted GPC3 gene sequences of small interfering RNA (siRNA) PGC-shRNA-GPC3 were restructured. Stable expression cell lines of siRNA were screened and established with the help of liposomes (lipofectamine(TM2000)) as carrier transfection of human liver cell lines. In order to validate siRNA interference efficiency, GPC3 siRNA mRNA expression was detected after transfection by using RT-PCR and Western blot. The absorbance value of the cells of blank group, untransfection group and transfection group, the cell cycle and cell apoptosis were calculated, and effects of GPC3 gene on Huh-7 cell proliferation and apoptosis were observed. In the liver cancer cell lines Huh-7, GPC3 gene showed high expression. PGC-shRNA-GPC3 recombinant plasmid was constructed successfully via sequencing validation. Stable recombinant plasmid transfected into liver cancer cell lines Huh-7 can obviously inhibit GPC3 mRNA expression level. The targeted GPC3 siRNA can effectively inhibit the expression of GPC3. Copyright © 2014 Hainan Medical College. Published by Elsevier B.V. All rights reserved.

  12. Targeted Nanomedicine for Suppression of CD44 and Simultaneous Cell Death Induction in Ovarian Cancer: an Optimal Delivery of siRNA and Anticancer Drug

    PubMed Central

    Shah, Vatsal; Taratula, Oleh; Garbuzenko, Olga B.; Taratula, Olena R.; Rodriguez-Rodriguez, Lorna; Minko, Tamara

    2013-01-01

    Purpose: The proposed project is aimed at enhancing the efficiency of epithelial ovarian cancer treatment and reducing adverse side effects of chemotherapy using nanotechnology. Overexpression of the CD44 membrane receptor results in tumor initiation, growth, tumor stem cells specific behavior, development of drug resistance, and metastases. We hypothesize that a developed cancer targeted delivery system which combines CD44 siRNA with paclitaxel would successfully deliver its payload inside cancer cells, effectively induce cell death, and prevent metastases. Experimental Design: We synthesized, characterized, and tested a nanoscale-based drug delivery system containing a modified Polypropylenimine (PPI) dendrimer as a carrier; anticancer drug paclitaxel as a cell death inducer; a synthetic analog of luteinizing hormone-releasing hormone (LHRH) peptide as a tumor targeting moiety, and siRNA targeted to CD44 mRNA. The proposed NDDS was tested in vitro and in vivo using metastatic ovarian cancer cells isolated from patients with malignant ascites. Results: We found that in contrast to cells isolated from primary tumors, CD44 was highly overexpressed in metastatic cancer cells. Treatment with the proposed tumor-targeted nanoscale-based nucleic acid and drug delivery system led to the suppression of CD44 mRNA and protein, efficient induction of cell death, effective tumor shrinkage, and prevention of adverse side effects on healthy organs. Conclusion: We show a high therapeutic potential for combinatorial treatment of ovarian carcinoma with a novel drug delivery system that effectively transports siRNA targeting to CD44 mRNA simultaneously with cytotoxic agents. PMID:24036854

  13. Gene therapy for colorectal cancer by adenovirus-mediated siRNA targeting CD147 based on loss of the IGF2 imprinting system.

    PubMed

    Pan, Yuqin; He, Bangshun; Chen, Jie; Sun, Huiling; Deng, Qiwen; Wang, Feng; Ying, Houqun; Liu, Xian; Lin, Kang; Peng, Hongxin; Xie, Hongguang; Wang, Shukui

    2015-11-01

    Colorectal cancer (CRC) is one of the most common malignant tumors worldwide. Loss of imprinting (LOI) of the insulin-like growth factor 2 (IGF2) gene is an epigenetic abnormality phenomenon in CRC. Recently observed association of CRC with cluster of differentiation 147 (CD147) could provide a novel approach for gene therapy. In the present study, we investigated the feasibility of using adenovirus‑mediated siRNA targeting CD147 based on the IGF2 LOI system for targeted gene therapy of CRC. A novel adenovirus-mediated siRNA targeting CD147, rAd-H19-CD147mirsh, which was driven by the IGF2 imprinting system, was constructed. The results showed that the EGFP expression was detected only in the IGF2 LOI cell lines (HT-29 and HCT-8), but that no EGFP was produced in cell lines with maintenance of imprinting (MOI) (HCT116). Moreover, rAd-H19-CD147mirsh significantly inhibited the expression of CD147, decreased cell viability and invasive ability, and increased sensitivity to chemotherapeutic drugs only in the LOI cell lines in vitro. Furthermore, mice bearing HT-29 xenografted tumors, which received intratumoral administration of the rAd-H19-CD147mirsh, showed significantly reduced tumor growth and enhanced survival. We conclude that recombinant adenovirus-mediated siRNA targeting CD147 based on the IGF2 LOI system inhibited the growth of the LOI cells in vitro and in vivo, which would provide a novel approach for targeted CRC gene therapy.

  14. Improvement of corrosion protection property of Mg-alloy by DLC and Si-DLC coatings with PBII technique and multi-target DC-RF magnetron sputtering

    NASA Astrophysics Data System (ADS)

    Masami, Ikeyama; Setsuo, Nakao; Tsutomu, Sonoda; Junho, Choi

    2009-05-01

    Magnesium alloys have been considered as one of the most promising light weight materials with potential applications for automobile and aircraft components. Their poor corrosion resistance, however, has to date prevented wider usage. Diamond-like carbon (DLC) and silicon-incorporated DLC (Si-DLC) coatings are known to provide a high degree of corrosion protection, and hold accordingly promise for enhancing the corrosion resistance of the magnesium alloys. In this work we have studied the effect of coating conditions of DLC coatings as well as Si incorporation into coating on corrosion resistance, deposited onto AZ91 magnesium alloy substrates by plasma based ion implantation (PBII). The influences of a Ti interlayer beneath the DLC, Si-DLC and Ti incorporated DLC (Ti-DLC) coatings fabricated by multi-target direct-current radio-frequency (DC-RF) magnetron sputtering were also examined on both the adhesion strength and corrosion resistance of the materials. We have also examined the effect of the Si content in the Si-DLC coatings made by magnetron sputtering on the alloys' corrosion resistance. The results of potentiodynamic polarization measurements demonstrate that Si-DLC coating deposited by PBII exhibits the highest corrosion resistance in an aqueous 0.05 M NaCl solution. Although Ti layer is helpful in increasing adhesion between DLC coating and AZ91 substrate, it also influences adversely corrosion protection. The ozone treatment of the magnesium alloy's surface before the formation of coatings has been found to improve both adhesion strength and corrosion resistance.

  15. Effect of combination therapy of siRNA targeting growth hormone receptor and 5-fluorouracil in hepatic metastasis of colon cancer

    PubMed Central

    ZHOU, DONG; ZHANG, YI; LIANG, DAOMING; YUAN, YONG; ZENG, DEMIAO; CHEN, JIAYONG; YANG, JIE

    2015-01-01

    The aim of this study was to investigate the effects of small interfering RNA (siRNA) targeting human growth hormone receptor (hGHR) combined with 5-fluorouracil (5-FU) on the hepatic metastasis of colon cancer. The animal model of liver metastases using human SW480 colon cancer cells was established on BALB/c mice and the siRNA interfering plasmid targeting hGHR gene was constructed. The tumor-bearing mice were randomly divided into the saline control, plasmid, growth hormone (GH), 5-FU, 5-FU+plasmid and 5-FU+plasmid+GH groups. The liver metastasis in each group was observed. All the animals showed liver metastases and using siRNA-interfering plasmid treatment the incidence of liver metastases was significantly reduced in the tumor groups compared to the saline or GH group. The combined treatment of interfering plasmid and 5-FU slightly decreased the incidence of liver metastases in the tumor groups compared to the plasmid alone or 5-FU alone treatment, although the findings were not statistically significant. On the basis of the combination of interfering plasmid and 5-FU, the additional GH did not increase the incidence of liver metastases (P>0.05), but improved the weight loss of the mice (P<0.05) induced by the inhibition of GHR and toxicity of 5-FU. The present results showed that siRNA targeting hGHR is able to reduce the incidence of liver metastases of human SW480 colon cancer cells in mice. Thus, GHR may be important in tumor metastasis. PMID:26788158

  16. Microbeam synchrotron radiation diffraction study of a monocrystalline nickel-base turbine blade after service

    SciTech Connect

    Biermann, H.; Grossmann, B.V.; Mechsner, S.

    1997-11-01

    In turbine blades subjected to service, the hot regions near the leading and trailing edges are subjected to temperatures up to 1,100 C, whereas the regions near the cooling channels are subjected to temperatures of about 800 C. These temperature gradients cause strong inhomogeneities in the local thermal and mechanical loads. In the present paper a monocrystalline turbine blade of the nickel-base superalloy CMSX-6 with an orientation near [001] from a developmental turbine was investigated with high lateral resolution using a Bragg-Fresnel focusing optics in combination with synchrotron radiation at the European Synchrotron Radiation Facility (ESRF) in Grenoble. The blade had been exposed to service in so-called accelerated mission tests for several hundred hours in two test turbines. In the bulk of the material, a {gamma}/{gamma}{prime} raft structure perpendicular to the [001]-direction is observed. At the surface, the protective aluminide coating is visible. Between these two regions, a {gamma}{prime}-enriched zone exists.

  17. Labeling of stem cells with monocrystalline iron oxide for tracking and localization by magnetic resonance imaging

    PubMed Central

    Calzi, Sergio Li; Kent, David L.; Chang, Kyung-Hee; Padgett, Kyle R.; Afzal, Aqeela; Chandra, Saurav B.; Caballero, Sergio; English, Denis; Garlington, Wendy; Hiscott, Paul S.; Sheridan, Carl M.; Grant, Maria B.; Forder, John R.

    2013-01-01

    Precise localization of exogenously delivered stem cells is critical to our understanding of their reparative response. Our current inability to determine the exact location of small numbers of cells may hinder optimal development of these cells for clinical use. We describe a method using magnetic resonance imaging to track and localize small numbers of stem cells following transplantation. Endothelial progenitor cells (EPC) were labeled with monocrystalline iron oxide nanoparticles (MIONs) which neither adversely altered their viability nor their ability to migrate in vitro and allowed successful detection of limited numbers of these cells in muscle. MION-labeled stem cells were also injected into the vitreous cavity of mice undergoing the model of choroidal neovascularization, laser rupture of Bruch’s membrane. Migration of the MION-labeled cells from the injection site towards the laser burns was visualized by MRI. In conclusion, MION labeling of EPC provides a non-invasive means to define the location of small numbers of these cells. Localization of these cells following injection is critical to their optimization for therapy. PMID:19345699

  18. The effect of surface microstructure on the optical reflectance of monocrystalline silicon

    NASA Astrophysics Data System (ADS)

    Wang, Quanji; Zhou, Weidong; Chen, Fangfang; Yang, Ruizhao

    2016-12-01

    Surface texturing is an important technique used to enhance the light absorption by forming certain microstructures on silicon surface. In this article, four different microstructures, based on repeat units of triangles, perpendicular grooves, hexagons and parallel grooves respectively, were fabricated directly on the surface of monocrystalline silicon wafers by using femtosecond laser texturing technique. Compare to the silicon wafers that were not treated by laser, a significant decrease of light reflectance can be observed for those laser etched silicon surfaces. And the treated silicon surface with triangles texture was found to have the lowest relative reflectance of ∼20% in the wavelength range from 400 to 1000 nm, if the textured surfaces were irradiated using the same laser fabrication condition. In addition, the relative reflectance of laser etched silicon surfaces with similar repeat unit but different structural period was investigated as well. The results show that the relative reflectance of the treated surface increases along with the increase of structural period size. These results obtained here can provide a useful guide for fabricating silicon-based optoelectronic devices with a more excellent anti-reflective performance.

  19. Preparation of dendritic-like Ag crystals using monocrystalline silicon as template

    SciTech Connect

    Wei, Yanlin; Chen, Yashao; Ye, Linjing; Chang, Pengmei

    2011-06-15

    Research highlights: {yields} Template-assisted method for synthesis of dendritic silver. {yields} Unique dendritic silver structure with stems, branches, and leaves. {yields} The morphology of silver depends on silicon surface roughness. {yields} Both diffusion and oriented attachment dominating the dendritic structure formation. -- Abstract: Symmetric dendritic silver structures with controlled morphology were successfully synthesized by a solvothermal method with the assistance of monocrystalline silicon. The morphology and structure of the dendritic silver were characterized by transmission electron microscopy (TEM), powder X-ray diffraction (XRD), and scanning electron microscopy (SEM). It was found that the architecture of silver crystals could be controlled via simply adjusting the experiment parameters: AgNO{sub 3} concentration, reaction time and temperature. Moreover, structural characterizations suggested that the dendritic silver structures preferentially grew along (1 1 1) and (2 0 0) directions, leading to the formation of dendritic structures with 1-2 {mu}m in stem diameter and 10-50 {mu}m in length. Additionally, the formation process of the dendritic silver structures was studied, and a possible formation mechanism was proposed based on the experimental results.

  20. Design, fabrication and optical characterization of photonic crystal assisted thin film monocrystalline-silicon solar cells.

    PubMed

    Meng, Xianqin; Depauw, Valérie; Gomard, Guillaume; El Daif, Ounsi; Trompoukis, Christos; Drouard, Emmanuel; Jamois, Cécile; Fave, Alain; Dross, Frédéric; Gordon, Ivan; Seassal, Christian

    2012-07-02

    In this paper, we present the integration of an absorbing photonic crystal within a monocrystalline silicon thin film photovoltaic stack fabricated without epitaxy. Finite difference time domain optical simulations are performed in order to design one- and two-dimensional photonic crystals to assist crystalline silicon solar cells. The simulations show that the 1D and 2D patterned solar cell stacks would have an increased integrated absorption in the crystalline silicon layer would increase of respectively 38% and 50%, when compared to a similar but unpatterned stack, in the whole wavelength range between 300 nm and 1100 nm. In order to fabricate such patterned stacks, we developed an effective set of processes based on laser holographic lithography, reactive ion etching and inductively coupled plasma etching. Optical measurements performed on the patterned stacks highlight the significant absorption increase achieved in the whole wavelength range of interest, as expected by simulation. Moreover, we show that with this design, the angle of incidence has almost no influence on the absorption for angles as high as around 60°.

  1. EFFECT OF SHOCK COMPRESSION METHOD ON THE DEFECT SUBSTRUCTURE IN MONOCRYSTALLINE COPPER

    SciTech Connect

    Cao, B Y; Lassila, D H; Schneider, M S; Kad, B K; Huang, C X; Xu, Y B; Kalantar, D H; Remington, B A; Meyers, M A

    2005-09-23

    Monocrystalline copper samples with orientations of [001] and [221] were shocked at pressures ranging from 20 GPa to 60 GPa using two techniques: direct drive lasers and explosively driven flyer plates. The pulse duration for these techniques differed substantially: 40 ns for the laser experiments at 0.5 mm into the sample and 1.1 {approx} 1.4 {micro}s for the flyer-plate experiments at 5 mm into the sample. The residual microstructures were dependent on orientation, pressure, and shocking method. The much shorter pulse duration in the laser driven shock yielded microstructures closer to the ones generated at the shock front. For the flyer-plate experiments, the longer pulse duration allows shock-generated defects to reorganize into lower energy configurations. Calculations show that the post-shock cooling for the laser driven shock is 10{sup 3} {approx} 10{sup 4} faster than that for plate-impact shock, propitiating recovery and recrystallization conditions for the latter. At the higher pressure level, extensive recrystallization was observed in the plate-impact samples, while it was absent in the laser driven shock. An effect that is proposed to contribute significantly to the formation of recrystallized regions is the existence of micro-shear-bands, which increase the local temperature beyond the prediction from adiabatic compression.

  2. Effect of Shock Compression Method on the Defect Substructure in Monocrystalline Copper

    SciTech Connect

    Cao, B Y; Meyers, M A; Lassila, D H; Schneider, M S; Kad, B K; Huang, C X; Xu, Y B; Kalantar, D H; Remington, B A

    2005-02-17

    Monocrystalline copper samples with orientations of [001] and [221] were shocked at pressures ranging from 20 GPa to 60 GPa using two techniques: direct drive lasers and explosively driven flyer plates. The pulse duration for these techniques differed substantially: 2 ns for the laser experiments and 1.1-1.4 {micro}s for the flyer-plate experiments. The residual microstructures were dependent on orientation, pressure, and shocking method. The much shorter pulse duration in laser shock yielded recovery microstructures with no or limited dislocation motion. For the flyer-plate experiments, the longer pulse duration allow shock-generated defects to reorganize into lower energy configurations. Calculations show that the post shock cooling occurs in a time scale of 0.2 s for laser shock and 1000 s for plate-impact shock, propitiating recovery and recrystallization conditions for the latter. At the higher pressure level extensive recrystallization was observed in the plate-impact samples, while it was absent in laser shock. An effect that is proposed to contribute significantly to the formation of recrystallized regions is the existence of micro-shearbands, which increase the local temperature.

  3. Porous silicon carbide (SiC) semiconductor device

    NASA Technical Reports Server (NTRS)

    Shor, Joseph S. (Inventor); Kurtz, Anthony D. (Inventor)

    1994-01-01

    A semiconductor device employs at least one layer of semiconducting porous silicon carbide (SiC). The porous SiC layer has a monocrystalline structure wherein the pore sizes, shapes, and spacing are determined by the processing conditions. In one embodiment, the semiconductor device is a p-n junction diode in which a layer of n-type SiC is positioned on a p-type layer of SiC, with the p-type layer positioned on a layer of silicon dioxide. Because of the UV luminescent properties of the semiconducting porous SiC layer, it may also be utilized for other devices such as LEDs and optoelectronic devices.

  4. Deposition of copper coatings in a magnetron with liquid target

    SciTech Connect

    Tumarkin, A. V. Kaziev, A. V.; Kolodko, D. V.; Pisarev, A. A.; Kharkov, M. M.; Khodachenko, G. V.

    2015-12-15

    Copper coatings were deposited on monocrystalline Si substrates using a magnetron discharge with a liquid cathode in the metal vapour plasma. During the deposition, the bias voltage in the range from 0 V to–400 V was applied to the substrate. The prepared films were investigated by a scanning electron microscope, and their adhesive properties were studied using a scratch tester. It was demonstrated that the adhesion of the deposited films strongly depends on the bias voltage and varies in a wide range.

  5. The siRNA targeted to mdr1b and mdr1a mRNAs in vivo sensitizes murine lymphosarcoma to chemotherapy

    PubMed Central

    2010-01-01

    Background One of the main obstacles for successful cancer polychemotherapy is multiple drug resistance phenotype (MDR) acquired by tumor cells. Currently, RNA interference represents a perspective strategy to overcome MDR via silencing the genes involved in development of this deleterious phenotype (genes of ABC transporters, antiapoptotic genes, etc.). Methods In this study, we used the siRNAs targeted to mdr1b, mdr1a, and bcl-2 mRNAs to reverse the MDR of tumors and increase tumor sensitivity to chemotherapeutics. The therapy consisting in ex vivo or in vivo application of mdr1b/1a siRNA followed by cyclophosphamide administration was studied in the mice bearing RLS40 lymphosarcoma, displaying high resistance to a wide range of cytostatics. Results Our data show that a single application of mdr1b/1a siRNA followed by treatment with conventionally used cytostatics results in more than threefold decrease in tumor size as compared with the control animals receiving only cytostatics. Conclusions In perspective, mdr1b/1a siRNA may become a well-reasoned adjuvant tool in the therapy of MDR malignancies. PMID:20470373

  6. Targeted nanoconjugate co-delivering siRNA and tyrosine kinase inhibitor to KRAS mutant NSCLC dissociates GAB1-SHP2 post oncogene knockdown

    NASA Astrophysics Data System (ADS)

    Srikar, R.; Suresh, Dhananjay; Zambre, Ajit; Taylor, Kristen; Chapman, Sarah; Leevy, Matthew; Upendran, Anandhi; Kannan, Raghuraman

    2016-08-01

    A tri-block nanoparticle (TBN) comprising of an enzymatically cleavable porous gelatin nanocore encapsulated with gefitinib (tyrosine kinase inhibitor (TKI)) and surface functionalized with cetuximab-siRNA conjugate has been synthesized. Targeted delivery of siRNA to undruggable KRAS mutated non-small cell lung cancer cells would sensitize the cells to TKI drugs and offers an efficient therapy for treating cancer; however, efficient delivery of siRNA and releasing it in cytoplasm remains a major challenge. We have shown TBN can efficiently deliver siRNA to cytoplasm of KRAS mutant H23 Non-Small Cell Lung Cancer (NSCLC) cells for oncogene knockdown; subsequently, sensitizing it to TKI. In the absence of TKI, the nanoparticle showed minimal toxicity suggesting that the cells adapt a parallel GAB1 mediated survival pathway. In H23 cells, activated ERK results in phosphorylation of GAB1 on serine and threonine residues to form GAB1-p85 PI3K complex. In the absence of TKI, knocking down the oncogene dephosphorylated ERK, and negated the complex formation. This event led to tyrosine phosphorylation at Tyr627 domain of GAB1 that regulated EGFR signaling by recruiting SHP2. In the presence of TKI, GAB1-SHP2 dissociation occurs, leading to cell death. The outcome of this study provides a promising platform for treating NSCLC patients harboring KRAS mutation.

  7. Targeted nanoconjugate co-delivering siRNA and tyrosine kinase inhibitor to KRAS mutant NSCLC dissociates GAB1-SHP2 post oncogene knockdown

    PubMed Central

    Srikar, R.; Suresh, Dhananjay; Zambre, Ajit; Taylor, Kristen; Chapman, Sarah; Leevy, Matthew; Upendran, Anandhi; Kannan, Raghuraman

    2016-01-01

    A tri-block nanoparticle (TBN) comprising of an enzymatically cleavable porous gelatin nanocore encapsulated with gefitinib (tyrosine kinase inhibitor (TKI)) and surface functionalized with cetuximab-siRNA conjugate has been synthesized. Targeted delivery of siRNA to undruggable KRAS mutated non-small cell lung cancer cells would sensitize the cells to TKI drugs and offers an efficient therapy for treating cancer; however, efficient delivery of siRNA and releasing it in cytoplasm remains a major challenge. We have shown TBN can efficiently deliver siRNA to cytoplasm of KRAS mutant H23 Non-Small Cell Lung Cancer (NSCLC) cells for oncogene knockdown; subsequently, sensitizing it to TKI. In the absence of TKI, the nanoparticle showed minimal toxicity suggesting that the cells adapt a parallel GAB1 mediated survival pathway. In H23 cells, activated ERK results in phosphorylation of GAB1 on serine and threonine residues to form GAB1-p85 PI3K complex. In the absence of TKI, knocking down the oncogene dephosphorylated ERK, and negated the complex formation. This event led to tyrosine phosphorylation at Tyr627 domain of GAB1 that regulated EGFR signaling by recruiting SHP2. In the presence of TKI, GAB1-SHP2 dissociation occurs, leading to cell death. The outcome of this study provides a promising platform for treating NSCLC patients harboring KRAS mutation. PMID:27530552

  8. Gradient nanostructured coatings obtained by magnetron sputtering of a multiphase AlN-TiB2-TiSi2 target

    NASA Astrophysics Data System (ADS)

    Pogrebnjak, A. D.; Pshyk, A. V.; Coy, E.; Zaleski, K.; Peplinska, B.; Nowaczyk, G.; Kupchishin, A. I.; Beresnev, V. M.; Kassenova, L. G.; Kravchenko, Ya. O.

    2016-10-01

    The preparation and analysis of gradient nanostructured coatings obtained by the method of the magnetron sputtering of a multiphase composite AlN-TiB2-TiSi2 target are described. The structure and phase and elemental compositions have been investigated by the methods of X-ray diffraction (XRD), atomic force microscopy (AFM), and electron microscopy (SEM and TEM, with energy-dispersive analysis). The mechanical properties of coatings were characterized by the method of nanoindentation. The coating formed consisted of three layers different in the elemental composition and structure, which determined its mechanical properties. The formation of structurally inhomogeneous coating is explained by the fact that the target to be sputtered consisted of three different components (AlN, 50 wt %; TiB2, 35 wt %; TiSi2, 15 wt %) inhomogeneously distributed over the volume of the target. The influence of different processes that occur upon the sputtering of multiphase targets by ions of inert gases on the formation of nanocomposite coatings with a gradient structure is discussed.

  9. Multilayered polyion complexes with dissolvable silica layer covered by controlling densities of cRGD-conjugated PEG chains for cancer-targeted siRNA delivery.

    PubMed

    Naito, Mitsuru; Azuma, Ryota; Takemoto, Hiroyasu; Hori, Mao; Yoshinaga, Naoto; Osawa, Shigehito; Kamegawa, Rimpei; Kim, Hyun Jin; Ishii, Takehiko; Nishiyama, Nobuhiro; Miyata, Kanjiro; Kataoka, Kazunori

    2017-03-12

    Surface functionalization of nanoparticles is a crucial factor for nanoparticle-mediated drug and nucleic acid delivery. Particularly, the density of targeting ligands on nanoparticle significantly affects the affinity of nanoparticles to specific cellular surface (or receptor) through the multivalent binding effect. Herein, multilayered polyion complexes (mPICs) are prepared to possess varying densities of cyclic RGD peptide (cRGD) ligands for cancer-targeted small interfering RNA (siRNA) delivery. A template PIC is first prepared by mixing siRNAs with homo catiomers of N-substituted polyaspartamide bearing tetraethylenepentamine (PAsp(TEP)) in aqueous solution, followed by silica-coating through silicate condensation reaction. Then, silica-coated PICs (sPICs) are further covered with block catiomers of PAsp(TEP) and poly(ethylene glycol) (PEG) equipped with cRGD ligand. Successful preparation of targeted mPICs is confirmed from the changes in size and ζ-potential and the elemental analysis by transmission electron microscopy. Notably, the number of cRGD ligands per mPIC is regulated by altering the silicate concentration upon preparation of sPICs, which is confirmed by fluorescence correlation spectroscopy using fluorescent-labeled block catiomers. Ultimately, the targeted mPICs with a higher number of cRGD ligands demonstrate more efficient cellular uptake in cultured cancer cells, leading to enhanced gene silencing activity.

  10. Tailoring Lipid and Polymeric Nanoparticles as siRNA Carriers towards the Blood-Brain Barrier - from Targeting to Safe Administration.

    PubMed

    Gomes, Maria João; Fernandes, Carlos; Martins, Susana; Borges, Fernanda; Sarmento, Bruno

    2017-03-01

    Blood-brain barrier is a tightly packed layer of endothelial cells surrounding the brain that acts as the main obstacle for drugs enter the central nervous system (CNS), due to its unique features, as tight junctions and drug efflux systems. Therefore, since the incidence of CNS disorders is increasing worldwide, medical therapeutics need to be improved. Consequently, aiming to surpass blood-brain barrier and overcome CNS disabilities, silencing P-glycoprotein as a drug efflux transporter at brain endothelial cells through siRNA is considered a promising approach. For siRNA enzymatic protection and efficient delivery to its target, two different nanoparticles platforms, solid lipid (SLN) and poly-lactic-co-glycolic (PLGA) nanoparticles were used in this study. Polymeric PLGA nanoparticles were around 115 nm in size and had 50 % of siRNA association efficiency, while SLN presented 150 nm and association efficiency close to 52 %. Their surface was functionalized with a peptide-binding transferrin receptor, in a site-oriented manner confirmed by NMR, and their targeting ability against human brain endothelial cells was successfully demonstrated by fluorescence microscopy and flow cytometry. The interaction of modified nanoparticles with brain endothelial cells increased 3-fold compared to non-modified lipid nanoparticles, and 4-fold compared to non-modified PLGA nanoparticles, respectively. These nanosystems, which were also demonstrated to be safe for human brain endothelial cells, without significant cytotoxicity, bring a new hopeful breath to the future of brain diseases therapies.

  11. Gas pressure atmosphere annealing: A novel method for the preparation of SiC nanowires

    NASA Astrophysics Data System (ADS)

    Zhang, X.; Zhong, B.; Liu, L.; Huang, X.; Wen, G.; Huang, Y.; Bollmann, J.

    2016-04-01

    Silicon carbide nanowires were fabricated by gas pressure annealing of SiOC nanocomposite powders, which were synthesized by pyrolysis of a SiO2 - sucrose gel. The reaction was carried out in an atmosphere sintering furnace without any additives. The nanowires have pronounced homogenous diameters smaller than 100 nm and lengths of up to several millimetres. The X-ray diffraction pattern indicates the formation of the β-SiC phase and transmission electron microscopy analysis show the monocrystalline structure of the nanowires.

  12. A general approach to prepare conjugated polymer dot embedded silica nanoparticles with a SiO2@CP@SiO2 structure for targeted HER2-positive cellular imaging

    NASA Astrophysics Data System (ADS)

    Geng, Junlong; Liu, Jie; Liang, Jing; Shi, Haibin; Liu, Bin

    2013-08-01

    We report on a one-step synthesis of conjugated polymer (CP) embedded silica nanoparticles (NPs) with a SiO2@CP@SiO2 structure by combination of a precipitation method and a modified Stöber approach. Four types of CPs are employed to demonstrate the versatility of the developed strategy, yielding fluorescent silica NPs with emission across the visible spectrum. Field emission transmission electron microscopy investigation reveals that the entanglement between hydrophobic CPs and the aminopropyl groups of 3-aminopropyl triethoxysilane contributes to the successful encapsulation of CPs into a silica matrix. The synthesized NPs exhibit excellent physical stability and good photostability. In addition, they have amine groups on surfaces, which benefit further conjugation for biological applications. Through reaction with a peptide (GGHAHFG) that is specific to the HER2 receptor, the synthesized NPs have been successfully applied for targeted cellular imaging of HER2-overexpressed SKBR-3 breast cancer cells. Along with its high quantum yield and benign biocompatibility, the developed CP embedded silica NPs have great potential for applications in biological imaging.We report on a one-step synthesis of conjugated polymer (CP) embedded silica nanoparticles (NPs) with a SiO2@CP@SiO2 structure by combination of a precipitation method and a modified Stöber approach. Four types of CPs are employed to demonstrate the versatility of the developed strategy, yielding fluorescent silica NPs with emission across the visible spectrum. Field emission transmission electron microscopy investigation reveals that the entanglement between hydrophobic CPs and the aminopropyl groups of 3-aminopropyl triethoxysilane contributes to the successful encapsulation of CPs into a silica matrix. The synthesized NPs exhibit excellent physical stability and good photostability. In addition, they have amine groups on surfaces, which benefit further conjugation for biological applications. Through

  13. Triple-Layered pH-Responsive Micelleplexes Loaded with siRNA and Cisplatin Prodrug for NF-Kappa B Targeted Treatment of Metastatic Breast Cancer

    PubMed Central

    Yu, Haijun; Guo, Chengyue; Feng, Bing; Liu, Jianping; Chen, Xianzhi; Wang, Dangge; Teng, Lesheng; Li, Youxin; Yin, Qi; Zhang, Zhiwen; Li, Yaping

    2016-01-01

    The combination of chemotherapy and RNA interference is a promising approach for efficient cancer therapy. However, the success of such a strategy is hampered by the lack of suitable vectors to coordinate small interfering RNA (siRNA) and chemotherapeutic drug into one single platform. We herein report a novel triple-layered pH-responsive micelleplex loading siRNA and alkylated cisplatin prodrug for NF-Kappa B targeted treatment of metastatic breast cancer. The micelles were self-assembled from poly(ethylene glycol)-block-poly(aminolated glycidyl methacrylate)-block-poly(2-(diisopropyl amino) ethyl methacrylate) (PEG-b-PAGA-b-PDPA) triblock copolymers. At pH 7.4, the cisplatin prodrug was encapsulated in the hydrophobic PDPA core and siRNA was loaded on the positively charged PAGA interlayer to form the micelleplexes. The PEG corona can prevent protein absorption during blood circulation, minimize non-specific interaction with the reticuloendothelial system, and prolong the systemic circulation of the micelleplexes. The positively charged PAGA interlayer can facilitate deep tumor penetration of the micelleplexes, which, upon cellular uptake, are dissociated in the early endosomes to release anticancer drug payload due to protonation of the PDPA core. Using a 4T1 breast cancer model, we demonstrate that this novel micelleplex co-loaded with cisplatin prodrug and siRNA-p65 is able to simultaneously inhibit tumor growth and suppress distant metastasis of the cancer cells by downregulating NF-kappa B expression. The results reported in this study suggest that siRNA and anticancer drug co-delivery using pH-responsive micelleplexes is a promising strategy for efficient treatment of metastatic cancer. PMID:26722370

  14. Assessment of drug delivery and anticancer potentials of nanoparticles-loaded siRNA targeting STAT3 in lung cancer, in vitro and in vivo.

    PubMed

    Das, Jayeeta; Das, Sreemanti; Paul, Avijit; Samadder, Asmita; Bhattacharyya, Soumya Sundar; Khuda-Bukhsh, Anisur Rahman

    2014-03-21

    Activation of signal transducer and activator of transcription3 (STAT3) is a hallmark of several types of cancer. Failure to inhibit STAT3 expression by injection of siRNA for STAT3 directly to Balb/c mice led us to adopt alternative means. We formulated nanoparticle-based encapsulation of siRNA (NsiRNA) with polyethylenimine (PEI) and poly(lactide-co-glycolide) (PLGA) and characterized them. The siRNA treated and NsiRNA-treated cells were subjected separately to different assay systems. We also checked if NsiRNA could cross the blood brain barrier (BBB). Cell viability reduced dramatically in A549 cells after NsiRNA administration (23.89% at 24 h), thereby implicating considerable silencing of STAT3 by NsiRNA, but not after siRNA administration. Compared to controls, a significant decrease in expression of IL-6 and the angiogenic factor (VEGF) and increase in Caspase 3 activity was observed with corresponding regression in tumor growth in mice treated with NsiRNA. NsiRNA induced apoptosis of cells and arrested cells at G1/G0 stage, both in vitro and in vivo. Apoptosis was also verified by Annexin-V-FITC/Propidium-iodide staining. NsiRNA could cross blood brain barrier. Overall results revealed PEI-PLGA to be a promising carrier for delivery of siRNA targeting STAT3 expression, which can be utilized as an effective strategy for cancer therapy. Copyright © 2014 Elsevier Ireland Ltd. All rights reserved.

  15. Triple-Layered pH-Responsive Micelleplexes Loaded with siRNA and Cisplatin Prodrug for NF-Kappa B Targeted Treatment of Metastatic Breast Cancer.

    PubMed

    Yu, Haijun; Guo, Chengyue; Feng, Bing; Liu, Jianping; Chen, Xianzhi; Wang, Dangge; Teng, Lesheng; Li, Youxin; Yin, Qi; Zhang, Zhiwen; Li, Yaping

    2016-01-01

    The combination of chemotherapy and RNA interference is a promising approach for efficient cancer therapy. However, the success of such a strategy is hampered by the lack of suitable vectors to coordinate small interfering RNA (siRNA) and chemotherapeutic drug into one single platform. We herein report a novel triple-layered pH-responsive micelleplex loading siRNA and alkylated cisplatin prodrug for NF-Kappa B targeted treatment of metastatic breast cancer. The micelles were self-assembled from poly(ethylene glycol)-block-poly(aminolated glycidyl methacrylate)-block-poly(2-(diisopropyl amino) ethyl methacrylate) (PEG-b-PAGA-b-PDPA) triblock copolymers. At pH 7.4, the cisplatin prodrug was encapsulated in the hydrophobic PDPA core and siRNA was loaded on the positively charged PAGA interlayer to form the micelleplexes. The PEG corona can prevent protein absorption during blood circulation, minimize non-specific interaction with the reticuloendothelial system, and prolong the systemic circulation of the micelleplexes. The positively charged PAGA interlayer can facilitate deep tumor penetration of the micelleplexes, which, upon cellular uptake, are dissociated in the early endosomes to release anticancer drug payload due to protonation of the PDPA core. Using a 4T1 breast cancer model, we demonstrate that this novel micelleplex co-loaded with cisplatin prodrug and siRNA-p65 is able to simultaneously inhibit tumor growth and suppress distant metastasis of the cancer cells by downregulating NF-kappa B expression. The results reported in this study suggest that siRNA and anticancer drug co-delivery using pH-responsive micelleplexes is a promising strategy for efficient treatment of metastatic cancer.

  16. Targeting L1 cell adhesion molecule expression using liposome-encapsulated siRNA suppresses prostate cancer bone metastasis and growth

    PubMed Central

    Sung, Shian-Ying; Petros, John A.; Wu, Hsi-Chin; Zeng, Hong-Jie; Huang, Wei-Chien; Chung, Leland W. K.; Hsieh, Chia-Ling

    2014-01-01

    The L1 cell adhesion molecule (L1CAM) has been implicated in tumor progression of many types of cancers, but its role in prostate cancer and its application in targeted gene therapy have not been investigated. Herein, we demonstrated that the L1CAM was expressed in androgen-insensitive and highly metastatic human prostate cancer cell lines. The correlation between L1CAM expression and prostate cancer metastasis was also validated in serum samples of prostate cancer patients. Knockdown of L1CAM expression in prostate cancer cells by RNA interference significantly decreased their aggressive behaviors, including colony formation, migration and invasion in vitro, and tumor formation in a metastatic murine model. These anti-malignant phenotypes of L1CAM-knockdown cancer cells were accompanied by G0/G1 cell cycle arrest and suppression of matrix metalloproteinase (MMP)-2 and MMP-9 expression and nuclear factor NF-κB activation. In vivo targeting of L1CAM expression using liposome-encapsulated L1CAM siRNAs effectively inhibited prostate cancer growth in mouse bone, which was associated with decreased L1CAM expression and cell proliferation by tumor cells. These results provide the first evidence for L1CAM being a major contributor to prostate cancer metastasis and translational application of siRNA-based L1CAM-targeted therapy. PMID:25294816

  17. In vitro and in vivo targeting imaging of pancreatic cancer using a Fe3O4@SiO2 nanoprobe modified with anti-mesothelin antibody

    PubMed Central

    Liu, Fang; Le, Wenjun; Mei, Tianxiao; Wang, Tiegong; Chen, Luguang; Lei, Yi; Cui, Shaobin; Chen, Bingdi; Cui, Zheng; Shao, Chengwei

    2016-01-01

    Pancreatic cancer is a highly malignant disease with a 5-year survival rate <5% mainly due to lack of early diagnosis and effective therapy. In an effort to improve the early diagnostic rate of pancreatic cancer, a nanoprobe Fe3O4@SiO2 modified with anti-mesothelin antibody (A-MFS) was prepared to target cells and tumor tissues highly expressing mesothelin in vitro (human pancreatic cancer cell line SW1990) and in vivo (subcutaneously transplanted tumors) studies. The A-MFS probe was successfully prepared and was spherical and uniform with a hydrodynamic diameter between 110 and 130 nm. Cell Counting Kit-8 testing indicated that A-MFS was nontoxic in vitro and in vivo studies. The in vitro study showed that the A-MFS probe specifically targeted SW1990 cells with high mesothelin expression. The in vivo study was conducted in Siemens 3.0 T magnetic resonance imaging. The average T2-weighted signal values of the xenografts were 966.533±31.56 before injecting A-MFS and 691.133±56.84 before injecting saline solution. After injection of 0.1 mL A-MFS via nude mouse caudal vein for 2.5 hours, the average T2-weighted signal of the xenograft decreased by 342.533±42.6. The signal value decreased by −61.233±33.9 and −58.7±19.4 after injection of the saline and Fe3O4@SiO2. The decrease of tumor signal by A-MFS was much more significant than that by saline and Fe3O4@SiO2 (P<0.05). The results demonstrated the high stability and nontoxicity of A-MFS, which effectively targeted pancreatic cancer in vitro and in vivo. A-MFS is a promising agent for diagnosis of pancreatic cancer. PMID:27274243

  18. Rational Design of GO-Modified Fe3O4/SiO2 Nanoparticles with Combined Rhenium-188 and Gambogic Acid for Magnetic Target Therapy.

    PubMed

    Yang, Yuxiang; Liu, Yicheng; Cheng, Chao; Shi, Haowei; Yang, Huan; Yuan, Hongming; Ni, Chaoying

    2017-08-30

    Peanutlike magnetic-fluorescent Fe3O4/SiO2 nanoparticles, with an effective dynamic diameter of 180 nm, were synthesized via EuO(+) doping and coupling of two Fe3O4 cores and reassembling through the solvothermal process. Spherical pure Fe3O4/SiO2 nanoparticles with an effective dynamic diameter of 230 nm were also prepared for comparison. We designed graphene oxide (GO)-modified core-shell Fe3O4/SiO2 nanoparticles as a nanocarrier for loading gambogic acid (GA) following labeling with radioisotope rhenium-188. We also performed GA loading and releasing on GA-loaded magnetic nanoparticles, in vivo biodistribution, and magnetic drug targeting therapy experiments. Results indicated that the GA-loaded magnetic nanoparticles demonstrate a clear pH-dependent drug release behavior, having a higher release rate in acidic environments. The in vivo biodistribution of the magnetic nanoparticles has morphologic dependency, and the peanutlike nanoparticles (PN-Fe3O4) tend to accumulate more in the spleen, lung, and liver than in the spherical nanoparticles (S-Fe3O4). The targeted therapy showed a higher efficacy of PN-Fe3O4 in inhibiting tumor cell growth than the nontargeted therapy. The polyethyleneimine (PEI) grafting of PN-Fe3O4 with amide bond was also designed to find an effective active targeting antitumor agent considering the fact that the PEI-GO conjugate has a higher GA load efficiency and the convergence effect.

  19. Photoionization of monocrystalline CVD diamond irradiated with ultrashort intense laser pulse

    NASA Astrophysics Data System (ADS)

    Lagomarsino, Stefano; Sciortino, Silvio; Obreshkov, Boyan; Apostolova, Tzveta; Corsi, Chiara; Bellini, Marco; Berdermann, Eleni; Schmidt, Christian J.

    2016-02-01

    Direct laser writing of conductive paths in synthetic diamond is of interest for implementation in radiation detection and clinical dosimetry. Unraveling the microscopic processes involved in laser irradiation of diamond below and close to the graphitization threshold under the same conditions as the experimental procedure used to produce three-dimensional devices is necessary to tune the laser parameters to optimal results. To this purpose a transient currents technique has been used to measure laser-induced current signals in monocrystalline diamond detectors in a wide range of laser intensities and at different bias voltages. The current transients vs time and the overall charge collected have been compared with theoretical simulations of the carrier dynamics along the duration and after the conclusion of the 30 fs laser pulse. The generated charge has been derived from the collected charge by evaluation of the lifetime of the carriers. The plasma volume has also been evaluated by measuring the modified region. The theoretical simulation has been implemented in the framework of the empirical pseudopotential method extended to include time-dependent couplings of valence electrons to the radiation field. The simulation, in the low-intensity regime, I ˜1 TW /cm2 , predicts substantial deviation from the traditional multiphoton ionization, due to nonperturbative effects involving electrons from degenerate valence bands. For strong field with intensity of about 50 TW /cm2, nonadiabatic effects of electron-hole pair excitation become prominent with high carrier densities eventually causing the optical breakdown of diamond. The comparison of theoretical prediction with experimental data of laser-generated charge vs laser energy density yields a good quantitative agreement over six orders of magnitude. At the highest intensities the change of slope in the trend is explained taking into account the dependence of the optical parameters and the carrier mobility on plasma

  20. siRNA targeting vaccinia virus double-stranded RNA binding protein [E3L] exerts potent antiviral effects.

    PubMed

    Dave, Rajnish S; McGettigan, James P; Qureshi, Tazeen; Schnell, Matthias J; Nunnari, Giuseppe; Pomerantz, Roger J

    2006-05-10

    The Vaccinia virus gene, E3L, encodes a double-stranded RNA [dsRNA]-binding protein. We hypothesized that, owing to the critical nature of dsRNA in triggering host innate antiviral responses, E3L-specific small-interfering RNAs [siRNAs] should be effective antiviral agents against pox viruses, for which Vaccinia virus is an appropriate surrogate. In this study, we have utilized two human cell types, namely, HeLa and 293T, one which responds to interferon [IFN]-beta and the other produces and responds to IFN-beta, respectively. The antiviral effects were equally robust in HeLa and 293T cells. However, in the case of 293T cells, several distinct features were observed, when IFN-beta is activated in these cells. Vaccinia virus replication was inhibited by 97% and 98% as compared to control infection in HeLa and 293T cells transfected with E3L-specific siRNAs, respectively. These studies demonstrate the utility of E3L-specific siRNAs as potent antiviral agents for small pox and related pox viruses.

  1. Selective epitaxial growth of graphene on SiC

    NASA Astrophysics Data System (ADS)

    Camara, N.; Rius, G.; Huntzinger, J.-R.; Tiberj, A.; Mestres, N.; Godignon, P.; Camassel, J.

    2008-09-01

    We present a method of selective epitaxial growth of few layers graphene (FLG) on a "prepatterned" silicon carbide (SiC) substrate. The methods involves, successively, the sputtering of a thin aluminium nitride (AlN) layer on top of a monocrystalline SiC substrate and, then, patterning it with e-beam lithography and wet etching. The sublimation of few atomic layers of Si from the SiC substrate occurs only through the selectively etched AlN layer. The presence of the Raman G-band at ˜1582cm-1 in the AlN-free areas is used to validate the concept. It gives absolute evidence of selective FLG growth.

  2. Selective epitaxial growth of graphene on SiC

    SciTech Connect

    Camara, N.; Rius, G.; Godignon, P.; Huntzinger, J.-R.; Tiberj, A.; Camassel, J.

    2008-09-22

    We present a method of selective epitaxial growth of few layers graphene (FLG) on a ''prepatterned'' silicon carbide (SiC) substrate. The methods involves, successively, the sputtering of a thin aluminium nitride (AlN) layer on top of a monocrystalline SiC substrate and, then, patterning it with e-beam lithography and wet etching. The sublimation of few atomic layers of Si from the SiC substrate occurs only through the selectively etched AlN layer. The presence of the Raman G-band at {approx}1582 cm{sup -1} in the AlN-free areas is used to validate the concept. It gives absolute evidence of selective FLG growth.

  3. Study of Nickel Silicide as a Copper Diffusion Barrier in Monocrystalline Silicon Solar Cells

    SciTech Connect

    Kale, Abhijit; Beese, Emily; Saenz, Theresa; Warren, Emily; Nemeth, William; Young, David; Marshall, Alexander; Florent, Karine; Kurinec, Santosh K.; Agarwal, Sumit; Stradins, Pauls

    2016-11-21

    NiSi as a conductive diffusion barrier to silicon has been studied. We demonstrate that the NiSi films formed using the single step annealing process are as good as the two step process using XRD and Raman. Quality of NiSi films formed using e-beam Ni and electroless Ni process has been compared. Incomplete surface coverage and presence of constituents other than Ni are the main challenges with electroless Ni. We also demonstrate that Cu reduces the thermal stability of NiSi films. The detection of Cu has proven to be difficult due to temperature limitations.

  4. siRNA screen for genes that affect Junín virus entry uncovers voltage-gated calcium channels as a therapeutic target

    PubMed Central

    Lavanya, Madakasira; Cuevas, Christian D.; Thomas, Monica; Cherry, Sara; Ross, Susan R.

    2014-01-01

    New world hemorrhagic fever arenaviruses infection of humans results in 15–30% mortality. We performed a high throughput siRNA screen with Junín virus glycoprotein-pseudotyped viruses to find potential host therapeutic targets. Voltage-gated calcium channels (VGCC) subunits, for which there are FDA-approved drugs, were identified in the screen. Knockdown of VGCC subunits or treatment with channel blockers diminished Junín virus-cell fusion and entry into cells and thereby decreased infection. Gabapentin, an FDA-approved drug used to treat neuropathic pain that targets the α2δ2 subunit, inhibited infection of mice by the Candid 1 vaccine strain of the virus. These findings demonstrate that VGCCs play a role in virus infection and have the potential to lead to therapeutic intervention of new world arenavirus infection. PMID:24068738

  5. Intensity analysis of XPS spectra to determine oxide uniformity - Application to SiO2/Si interfaces

    NASA Technical Reports Server (NTRS)

    Vasquez, R. P.; Grunthaner, F. J.

    1980-01-01

    A simple method of determining oxide uniformity is derived which requires no knowlege of film thickness, escape depth, or film composition. The method involves only the measurement of oxide and substrate intensities and is illustrated by analysis of XPS spectral data for thin SiO2 films grown both thermally and by low-temperature chemical vapor deposition on monocrystalline Si. A region 20-30 A thick is found near the SiO2/Si interface on thermally oxidized samples which has an inelastic mean free path 35% less than that found in the bulk oxide. This is interpreted as being due to lattice mismatch resulting in a strained region which is structurally, but not stoichiometrically, distinct from the bulk oxide.

  6. Intensity analysis of XPS spectra to determine oxide uniformity - Application to SiO2/Si interfaces

    NASA Technical Reports Server (NTRS)

    Vasquez, R. P.; Grunthaner, F. J.

    1980-01-01

    A simple method of determining oxide uniformity is derived which requires no knowlege of film thickness, escape depth, or film composition. The method involves only the measurement of oxide and substrate intensities and is illustrated by analysis of XPS spectral data for thin SiO2 films grown both thermally and by low-temperature chemical vapor deposition on monocrystalline Si. A region 20-30 A thick is found near the SiO2/Si interface on thermally oxidized samples which has an inelastic mean free path 35% less than that found in the bulk oxide. This is interpreted as being due to lattice mismatch resulting in a strained region which is structurally, but not stoichiometrically, distinct from the bulk oxide.

  7. Stress control of reactively sputtered thick NbN film on Si wafer changing the location of the substrate Si wafer against the Nb target on a magnetron cathode

    NASA Astrophysics Data System (ADS)

    Suzuki, Y.; Iguchi, N.; Adachi, K.; Hioki, T.; Ichiki, A.; Hsu, C.-W.; Kumagai, S.; Sasaki, M.; Motohiro, T.

    2017-07-01

    We have been developing a superconducting NbN thin film coil in a spiral trench on a Si-wafer using MEMS technology. Connecting the coils on the different wafers using waferbonding process, a cylindrical wafer stack is to be formed as a unit of a compact SMES. The critical current density of our NbN film was measured to be around 1100 A/mm2. We measured critical current I c of 47 mA for the previously fabricated coil of the film thickness t f = 0.5 μm. I c in the spiral coil increases with t f. However, if we make the NbN film thicker, the film is apt to have higher lateral force caused by tensile or compressive stress which can cause peeling of the film from the Si substrate. It is well known that the stress of the sputtered thin films can be controlled from tensile to compressive stress by controlling the bombardment of high energy particles including argon atoms backscattered from the target surface. Based on this knowledge, a specially designed sputter-deposition apparatus was fabricated in which the substrate can be located not only at the different target-to-substrate distances but also at several different lateral distances from the central axis of the target (off-axis lateral shift). Using this apparatus, various stress conditions could be realized which contributed to fabrication of thick NbN film spiral coil in the trench. The film stress was calculated from bending analysis of the substrate Si wafer by stylus method using Stoney’s formula. The maximum compressive stress of 2.5 GPa was measured. By an off-axis lateral shift, t f could be increased from 0.5 to 1 μm. By increasing sputtering gas pressure from 0.7 to 2 Pa, the compressive stress could be mitigated and t f could be further increased from 1.0 to 3 μm. Up to now, we measured I c of 220 mA for a NbN spiral coil at t f around 3 μm. More detailed adjustment of the deposition condition will bring further increase in t f, and hence I c into sight.

  8. Systemic co-delivery of doxorubicin and siRNA using nanoparticles conjugated with EGFR-specific targeting peptide to enhance chemotherapy in ovarian tumor bearing mice

    NASA Astrophysics Data System (ADS)

    Liu, C. W.; Lin, W. J.

    2013-10-01

    This aim of this study was to develop peptide-conjugated nanoparticles (NPs) for systemic co-delivery of siRNA and doxorubicin to enhance chemotherapy in epidermal growth factor receptor (EGFR) high-expressed ovarian tumor bearing mice. The active targeting NPs were prepared using heptapeptide-conjugated poly( d, l-lactic-co-glycolic acid)-poly(ethylene glycol). The particle sizes of peptide-free and peptide-conjugated NPs were 159.3 ± 32.5 and 184.0 ± 52.9 nm, respectively, with zeta potential -21.3 ± 3.8 and -15.3 ± 2.8 mV. The peptide-conjugated NPs uptake were more efficient in EGFR high-expressed SKOV3 cells than in EGFR low-expressed HepG2 cells due to heptapeptide specificity. The NPs were used to deliver small molecule anticancer drug (e.g., doxorubicin) and large molecule genetic agent (e.g., siRNA). The IC50 of doxorubicin-loaded peptide-conjugated NPs (0.09 ± 0.06 μM) was significantly lower than peptide-free NPs (5.72 ± 2.64 μM). The similar result was observed in siRNA-loaded NPs. The peptide-conjugated NPs not only served as a nanocarrier to efficiently deliver doxorubicin and siRNA to EGFR high-expressed ovarian cancer cells but also increased the intracellular accumulation of the therapeutic agents to induce assured anti-tumor growth effect in vivo.

  9. Target RNA-directed tailing and trimming purifies the sorting of endo-siRNAs between the two Drosophila Argonaute proteins.

    PubMed

    Ameres, Stefan L; Hung, Jui-Hung; Xu, Jia; Weng, Zhiping; Zamore, Phillip D

    2011-01-01

    In flies, 22-23-nucleotide (nt) microRNA duplexes typically contain mismatches and begin with uridine, so they bind Argonaute1 (Ago1), whereas 21-nt siRNA duplexes are perfectly paired and begin with cytidine, promoting their loading into Ago2. A subset of Drosophila endogenous siRNAs-the hairpin-derived hp-esiRNAs-are born as mismatched duplexes that often begin with uridine. These would be predicted to load into Ago1, yet accumulate at steady-state bound to Ago2. In vitro, such hp-esiRNA duplexes assemble into Ago1. In vivo, they encounter complementary target mRNAs that trigger their tailing and trimming, causing Ago1-loaded hp-esiRNAs to be degraded. In contrast, Ago2-associated hp-esiRNAs are 2'-O-methyl modified at their 3' ends, protecting them from tailing and trimming. Consequently, the steady-state distribution of esiRNAs reflects not only their initial sorting between Ago1 and Ago2 according to their duplex structure, length, and first nucleotide, but also the targeted destruction of the single-stranded small RNAs after their loading into an Argonaute protein.

  10. Multifunctional NaYF4:Yb, Er@mSiO2@Fe3O4-PEG nanoparticles for UCL/MR bioimaging and magnetically targeted drug delivery

    NASA Astrophysics Data System (ADS)

    Liu, Bei; Li, Chunxia; Ma, Ping'an; Chen, Yinyin; Zhang, Yuanxin; Hou, Zhiyao; Huang, Shanshan; Lin, Jun

    2015-01-01

    A low toxic multifunctional nanoplatform, integrating both mutimodal diagnosis methods and antitumor therapy, is highly desirable to assure its antitumor efficiency. In this work, we show a convenient and adjustable synthesis of multifunctional nanoparticles NaYF4:Yb, Er@mSiO2@Fe3O4-PEG (MFNPs) based on different sizes of up-conversion nanoparticles (UCNPs). With strong up-conversion fluorescence offered by UCNPs, superparamagnetism properties attributed to Fe3O4 nanoparticles and porous structure coming from the mesoporous SiO2 shell, the as-obtained MFNPs can be utilized not only as a contrast agent for dual modal up-conversion luminescence (UCL)/magnetic resonance (MR) bio-imaging, but can also achieve an effective magnetically targeted antitumor chemotherapy both in vitro and in vivo. Furthermore, the UCL intensity of UCNPs and the magnetic properties of Fe3O4 in the MFNPs were carefully balanced. Silica coating and further PEG modifying can improve the hydrophilicity and biocompatibility of the as-synthesized MFNPs, which was confirmed by the in vitro/in vivo biocompatibility and in vivo long-time bio-distributions tests. Those results revealed that the UCNPs based magnetically targeted drug carrier system we synthesized has great promise in the future for multimodal bio-imaging and targeted cancer therapy.A low toxic multifunctional nanoplatform, integrating both mutimodal diagnosis methods and antitumor therapy, is highly desirable to assure its antitumor efficiency. In this work, we show a convenient and adjustable synthesis of multifunctional nanoparticles NaYF4:Yb, Er@mSiO2@Fe3O4-PEG (MFNPs) based on different sizes of up-conversion nanoparticles (UCNPs). With strong up-conversion fluorescence offered by UCNPs, superparamagnetism properties attributed to Fe3O4 nanoparticles and porous structure coming from the mesoporous SiO2 shell, the as-obtained MFNPs can be utilized not only as a contrast agent for dual modal up-conversion luminescence (UCL

  11. High performance LWIR microbolometer with Si/SiGe quantum well thermistor and wafer level packaging

    NASA Astrophysics Data System (ADS)

    Roer, Audun; Lapadatu, Adriana; Bring, Martin; Wolla, Erik; Hohler, Erling; Kittilsland, Gjermund

    2011-11-01

    An uncooled microbolometer with peak responsivity in the long wave infrared region of the electromagnetic radiation is developed at Sensonor Technologies. It is a 384 x 288 focal plane array with a pixel pitch of 25μm, based on monocrystalline Si/SiGe quantum wells as IR sensitive material. The high sensitivity (TCR) and low 1/f noise are the main performance characteristics of the product. The frame rate is maximum 60Hz and the output interface is digital (LVDS). The quantum well thermistor material is transferred to the read-out integrated circuit (ROIC) by direct wafer bonding. The ROIC wafer containing the released pixels is bonded in vacuum with a silicon cap wafer, providing hermetic encapsulation at low cost. The resulting wafer stack is mounted in a standard ceramic package. In this paper the architecture of the pixels and the ROIC, the wafer packaging and the electro-optical measurement results are presented.

  12. A targeted siRNA screen identifies regulators of Cdc42 activity at the natural killer cell immunological synapse.

    PubMed

    Carlin, Leo M; Evans, Rachel; Milewicz, Hanna; Fernandes, Luis; Matthews, Daniel R; Perani, Michela; Levitt, James; Keppler, Melanie D; Monypenny, James; Coolen, Ton; Barber, Paul R; Vojnovic, Borivoj; Suhling, Klaus; Fraternali, Franca; Ameer-Beg, Simon; Parker, Peter J; Thomas, N Shaun B; Ng, Tony

    2011-11-29

    Natural killer (NK) cells kill tumor cells and virally infected cells, and an effective NK cell response requires processes, such as motility, recognition, and directional secretion, that rely on cytoskeletal rearrangement. The Rho guanosine triphosphatase (GTPase) Cdc42 coordinates cytoskeletal reorganization downstream of many receptors. The Rho-related GTPase from plants 1 (ROP1) exhibits oscillatory activation behavior at the apical plasma membrane of growing pollen tubes; however, a similar oscillation in Rho GTPase activity has so far not been demonstrated in mammalian cells. We hypothesized that oscillations in Cdc42 activity might occur within NK cells as they interact with target cells. Through fluorescence lifetime imaging of a Cdc42 biosensor, we observed that in live NK cells forming immunological synapses with target cells, Cdc42 activity oscillated after exhibiting an initial increase. We used protein-protein interaction networks and structural databases to identify candidate proteins that controlled Cdc42 activity, leading to the design of a targeted short interfering RNA screen. The guanine nucleotide exchange factors RhoGEF6 and RhoGEF7 were necessary for Cdc42 activation within the NK cell immunological synapse. In addition, the kinase Akt and the p85α subunit of phosphoinositide 3-kinase (PI3K) were required for Cdc42 activation, the periodicity of the oscillation in Cdc42 activity, and the subsequent polarization of cytotoxic vesicles toward target cells. Given that PI3Ks are targets of tumor therapies, our findings suggest the need to monitor innate immune function during the course of targeted therapy against these enzymes.

  13. Processing of LEU targets for {sup 99}Mo production: Dissolution of U{sub 3}Si{sub 2} targets by alkaline hydrogen peroxide

    SciTech Connect

    Buchholz, B.A.; Vandegrift, G.F.

    1995-09-01

    Low-enriched uranium silicide targets designed to recover fission product {sup 99}Mo were dissolved in alkaline hydrogen peroxide (H{sub 2}O{sub 2} plus NaOH) at about 90C. Sintering of matrix aluminum powder during irradiation and heat treatment retarded aluminum dissolution and prevented silicide particle dispersion. Gas evolved during dissolution is suspected to adhere to particles and block hydroxide ion contact with aluminum. Reduction of base concentrations from 5M to O.lM NaOH yielded similar silicide dissolution and peroxide destruction rates, simplifying later processing. Future work in particle dispersion enhancement, {sup 99}Mo separation, and waste disposal is also discussed.

  14. Flexible carbon nanotube/mono-crystalline Si thin-film solar cells

    PubMed Central

    2014-01-01

    Flexible heterojunction solar cells were fabricated from carbon nanotubes (CNTs) and mono-crystalline Si thin films at room temperature. The Si thin films with thickness less than 50 μm are prepared by chemically etching Si wafer in a KOH solution. The initial efficiency of the thin-film solar cell varies from approximately 3% to 5%. After doping with a few drops of 1 M HNO3, the efficiency increases to 6% with a short-circuit current density of 16.8 mA/cm2 and a fill factor of 71.5%. The performance of the solar cells depends on the surface state and thickness of Si thin films, as well as the interface of CNT/Si. The flexible CNT/Si thin-film solar cells exhibit good stability in bending-recovery cycles. PMID:25258617

  15. Mechanical properties, chemical analysis and evaluation of antimicrobial response of Si-DLC coatings fabricated on AISI 316 LVM substrate by a multi-target DC-RF magnetron sputtering method for potential biomedical applications

    NASA Astrophysics Data System (ADS)

    Bociaga, Dorota; Sobczyk-Guzenda, Anna; Szymanski, Witold; Jedrzejczak, Anna; Jastrzebska, Aleksandra; Olejnik, Anna; Jastrzebski, Krzysztof

    2017-09-01

    In this study silicon doped diamond-like carbon (Si-DLC) coatings were synthesized on two substrates: silicon and AISI 316LVM stainless steel using a multi-target DC-RF magnetron sputtering method. The Si content in the films ranged between 4 and 16 at.%, and was controlled by the electrical power applied in RF regime to Si cathode target. The character of the chemical bonds was revealed by FTIR analysis. With the addition of silicon the hydroxyl absorption (band in the range of 3200-3600 cm-1) increased what suggests more hydrophilic character of the coating. There were also observed significant changes in bonding of Si atoms. For low content of dopant, Si-O-Si bond system is predominant, while for the highest content of silicon there is an evidence of the shift to Si-C bonds in close proximity to methyl groups. The Raman spectroscopy revealed that the G peak position is shifted to a lower wavenumber and the ID/IG ratio decreased with increasing Si content, which indicates an increase in the C-sp3 content. Regardless of the coatings' composition, the improvement of hardness in comparison to pure substrate material (AISI 316 LVM) was observed. Although the reduction of the level of hardness from the level of 10.8 GPa for pure DLC to about 9.4 GPa for the silicon doped coatings was observed, the concomitant improvement of films adhesion with higher amount of Si was revealed. Although incorporation of the dopant to DLC coatings increases the number of E. coli cells which adhered to the examined surfaces, the microbial colonisation remains on the level of substrate material. The presented results prove the potential of Si-DLC coatings in biomedical applications from the point of view of their mechanical properties.

  16. Fabrication of Double Shell Targets with a Glass Inner Capsule Supported by SiO2 Aerogel for Shots on the Omega Laser in 2006

    SciTech Connect

    Bono, M; Bennett, D; Castro, C; Satcher, J; Poco, J; Brown, W; Martz, H; Teslich, N; Hibbard, R; Hamza, A; Amendt, P; Robey, H; Milovich, J; Wallace, R

    2006-10-26

    Indirectly driven double shell implosions are being investigated as a possible noncryogenic path to ignition on the National Ignition Facility. Lawrence Livermore National Laboratory has made several technological advances that have produced double shell targets that represent a significant improvement to previously fielded targets. The inner capsule is supported inside the ablator shell by SiO{sub 2} aerogel with a nominal density of 50 mg/cm{sup 3}. The aerogel is cast around the inner capsule and then machined concentric to it. The seamless sphere of aerogel containing the embedded capsule is then assembled between the two halves of the ablator shell. The concentricity between the two shells has been improved to less than 1.5 {micro}m. The ablator shell consists of two hemispherical shells that mate at a step joint that incorporates a gap with a nominal thickness of 0.1 {micro}m. Using a new flexure-based tool holder that precisely positions the diamond cutting tool on the diamond turning machine, step discontinuities on the inner surface of the ablator of less than 0.5 {micro}m have been achieved. New methods have been used to comprehensively characterize each of the targets using high-resolution x-ray imaging systems.

  17. Fabrication of Polyvalent Therapeutic RNA Nanoparticles for Specific Delivery of siRNA, Ribozyme and Drugs to Targeted Cells for Cancer Therapy

    PubMed Central

    Shu, Yi; Shu, Dan; Diao, Zhijuan; Shen, Guanxin; Guo, Peixuan

    2010-01-01

    Bacteriophage phi29 DNA packaging motor is geared by a six-pRNA ring. pRNA is able to form a multimeric complex and patterned superstructures via the interaction of two reengineered interlocking loops. This unique feature makes it an ideal polyvalent vehicle for nanomachine fabrication, pathogen detection, and the delivery of therapeutics. This report describes novel approaches for the fabrication of polyvalent therapeutic pRNA nanoparticles, especially tetramers for specific siRNA delivery to cancer cells and for the silencing of targeted genes. RNA 3-D design, circular permutation, folding energy alteration, and nucleotide modification were applied to generate stable RNA nanoparticles with low toxicity. Animal trials demonstrated the high efficiency of the polyvalent RNA nanoparticles in the prevention and treatment of cancer. Using such protein-free nanoparticles as therapeutic reagents would allow for long-term administration to avoid the induction of antibody due to repeated treatment for chronic diseases. PMID:21243099

  18. Targeted Dual pH-Sensitive Lipid ECO/siRNA Self-Assembly Nanoparticles Facilitate In Vivo Cytosolic sieIF4E Delivery and Overcome Paclitaxel Resistance in Breast Cancer Therapy.

    PubMed

    Gujrati, Maneesh; Vaidya, Amita M; Mack, Margaret; Snyder, Dayton; Malamas, Anthony; Lu, Zheng-Rong

    2016-11-01

    RNAi-mediated knockdown of oncogenes associated with drug resistance can potentially enhance the efficacy of chemotherapy. Here, we have designed and developed targeted dual pH-sensitive lipid-siRNA self-assembly nanoparticles, RGD-PEG(HZ)-ECO/siRNA, which can efficiently silence the oncogene, eukaryotic translation initiation factor 4E (eIF4E), and consequently resensitize triple-negative breast tumors to paclitaxel. The dual pH-sensitive function of these nanoparticles facilitates effective cytosolic siRNA delivery in cancer cells, both in vitro and in vivo. Intravenous injection of RGD-PEG(HZ)-ECO/siRNA nanoparticles (1.0 mg-siRNA/kg) results in effective gene silencing for at least one week in MDA-MB-231 tumors. In addition, treatment of athymic nude mice with RGD-PEG(HZ)-ECO/sieIF4E every 6 days for 6 weeks down-regulates the overexpression of eIF4E and resensitizes paclitaxel-resistant MDA-MB-231 tumors to paclitaxel, resulting in significant tumor regression at a low dose, with negligible side effects. Moreover, repeated injections of the RGD-PEG(HZ)-ECO/siRNA nanoparticles in immunocompetent mice result in minimal immunogenicity, demonstrating their safety and low toxicity. These multifunctional lipid/siRNA nanoparticles constitute a versatile platform of delivery of therapeutic siRNA for treating cancer and other human diseases. © 2016 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  19. PEGylation of the GALA Peptide Enhances the Lung-Targeting Activity of Nanocarriers That Contain Encapsulated siRNA.

    PubMed

    Santiwarangkool, Sarochin; Akita, Hidekata; Nakatani, Taichi; Kusumoto, Kenji; Kimura, Hiroki; Suzuki, Masaru; Nishimura, Masaharu; Sato, Yusuke; Harashima, Hideyoshi

    2017-09-01

    A α-helical GALA peptide (WEAALAEALAEALAEHLAEALAEALEALAA) has been found to possess dual functions: a pH-dependent inducer of endosomal escape, and a ligand that targets lung endothelium. In the present study, the flexibility of GALA was improved by modifying the edge with polyethylene glycol linker, to increase lung-targeting activity. We first investigated the uptake of the GALA-modified liposomes in which GALA was directly conjugated to the lipid (Cholesterol: GALA/Chol) or the phospholipid-PEG (GALA/PEG2000). The liposomes that were modified with GALA/PEG2000 (GALA/PEG2000-LPs) were taken up at a higher level by human lung endothelial cells (HMVEC-L), in comparison with particles that were modified with GALA/Chol (GALA/Chol-LPs). Small-interfering RNA-encapsulating liposomal-based nanocarriers (multifunctional envelope-type nano device: MEND) that were formulated with a vitamin E-scaffold SS-cleavable pH-activated lipid-like material, namely GALA/PEG2000-MENDssPalmE were also modified with GALA/PEG2000. Gene silencing activity in the lung endothelium was then evaluated against an endothelial marker; CD31. In comparison with the unmodified MENDssPalmE, GALA/PEG2000-MENDssPalmE exhibited a higher silencing activity in the lung. Optimization of GALA/PEG2000-MENDssPalmE resulted in silencing activity in the lung with an ED50 value of 0.21 mg/kg, while non-specific gene silencing in liver was marginal. Collectively, PEGylated GALA is a promising device for use in targeting the lung endothelium. Copyright © 2017 American Pharmacists Association®. Published by Elsevier Inc. All rights reserved.

  20. Nearly zero reflectance of nano-pyramids and dual-antireflection coating structure for monocrystalline silicon solar cells.

    PubMed

    Chang, Hyo Sik; Jung, Hyun-Chul

    2011-04-01

    The effect of two-step surface treatment on monocrystalline silicon solar cells was investigated. We changed the nanostructure on pyramidal surfaces by wet nano-texturing so that less light is reflected. The two-step nano-texturing process reduces the average reflectance to about 4% in the 300-1100 nm wavelength region. The use of an antireflection coating resulted in an effective reflectance of 1%. We found that the reflectance obtained by wet nano-texturing was lower than that obtained by conventional alkaline texturing. Thus, we can expect a further increase in the efficiency of silicon solar cells with two-step nano-texturing by a wet chemical process.

  1. Single heterojunction solar cells on exfoliated flexible ˜25 μm thick mono-crystalline silicon substrates

    NASA Astrophysics Data System (ADS)

    Saha, Sayan; Hilali, Mohamed M.; Onyegam, Emmanuel U.; Sarkar, Dabraj; Jawarani, Dharmesh; Rao, Rajesh A.; Mathew, Leo; Smith, Ryan S.; Xu, Dewei; Das, Ujjwal K.; Sopori, Bhushan; Banerjee, Sanjay K.

    2013-04-01

    Mono-crystalline silicon single heterojunction solar cells on flexible, ultra-thin (˜25 μm) substrates have been developed based on a kerf-less exfoliation method. Optical and electrical measurements demonstrate maintained structural integrity of these flexible substrates. Among several single heterojunction ˜25 μm thick solar cells fabricated with un-optimized processes, the highest open circuit voltage of 603 mV, short circuit current of 34.4 mA/cm2, and conversion efficiency of 14.9% are achieved separately on three different cells. Preliminary reliability test results that include thermal shock and highly accelerated stress tests are also shown to demonstrate compatibility of this technology for use in photovoltaic modules.

  2. Atomistic simulations of the effect of embedded hydrogen and helium on the tensile properties of monocrystalline and nanocrystalline tungsten

    NASA Astrophysics Data System (ADS)

    Chen, Zhe; Kecskes, Laszlo J.; Zhu, Kaigui; Wei, Qiuming

    2016-12-01

    Uniaxial tensile properties of monocrystalline tungsten (MC-W) and nanocrystalline tungsten (NC-W) with embedded hydrogen and helium atoms have been investigated using molecular dynamics (MD) simulations in the context of radiation damage evolution. Different strain rates have been imposed to investigate the strain rate sensitivity (SRS) of the samples. Results show that the plastic deformation processes of MC-W and NC-W are dominated by different mechanisms, namely dislocation-based for MC-W and grain boundary-based activities for NC-W, respectively. For MC-W, the SRS increases and a transition appears in the deformation mechanism with increasing embedded atom concentration. However, no obvious embedded atom concentration dependence of the SRS has been observed for NC-W. Instead, in the latter case, the embedded atoms facilitate GB sliding and intergranular fracture. Additionally, a strong strain enhanced He cluster growth has been observed. The corresponding underlying mechanisms are discussed.

  3. "Pop-in" and "pop-out" effect in monocrystalline silicon. A statistical investigation

    NASA Astrophysics Data System (ADS)

    Sidiropoulos, Alexandros D.; Harea, Evghenii; Konstantinidis, Avraam A.; Aifantis, Elias C.

    2017-04-01

    Pop-in and pop-out effects in silicon (Si) have long been known. They were evidenced in the indentation loading-unloading curves as a sudden displacement discontinuity. They consist in a sudden contraction (pop-in) or a sudden expansion (pop-out) of the material underneath the indenter in a short period of time and are attributed to Si phase transformations that take place during the nanoindentation procedure. In this paper, first we provide a statistic analysis of such pop-in/pop-out events depending on the maximum indentation load and second we examine the dependence of their appearance on the indentation loading-unloading rate.

  4. Apoptosis induced by a HIPK2 full-length-specific siRNA is due to off-target effects rather than prevalence of HIPK2-Δe8 isoform

    PubMed Central

    Di Rocco, Giuliana; Verdina, Alessandra; Gatti, Veronica; Virdia, Ilaria; Toietta, Gabriele; Todaro, Matilde; Stassi, Giorgio; Soddu, Silvia

    2016-01-01

    Small interfering RNAs (siRNAs) are widely used to study gene function and extensively exploited for their potential therapeutic applications. HIPK2 is an evolutionary conserved kinase that binds and phosphorylates several proteins directly or indirectly related to apoptosis. Recently, an alternatively spliced isoform skipping 81 nucleotides of exon 8 (Hipk2-Δe8) has been described. Selective depletion of Hipk2 full-length (Hipk2-FL) with a specific siRNA that spares the Hipk2-Δe8 isoform has been shown to strongly induce apoptosis, suggesting an unpredicted dominant-negative effect of Hipk2-FL over the Δe8 isoform. From this observation, we sought to take advantage and assessed the therapeutic potential of generating Hipk2 isoform unbalance in tumor-initiating cells derived from colorectal cancer patients. Strong reduction of cell viability was induced in vitro and in vivo by the originally described exon 8-specific siRNA, supporting a potential therapeutic application. However, validation analyses performed with additional exon8-specific siRNAs with different stabilities showed that all exon8-targeting siRNAs can induce comparable Hipk2 isoform unbalance but only the originally reported e8-siRNA promotes cell death. These data show that loss of viability does not depend on the prevalence of Hipk2-Δe8 isoform but it is rather due to microRNA-like off-target effects. PMID:26625198

  5. Enhancement of dendritic cell-based vaccine potency by anti-apoptotic siRNAs targeting key pro-apoptotic proteins in cytotoxic CD8(+) T cell-mediated cell death.

    PubMed

    Kim, Jin Hee; Kang, Tae Heung; Noh, Kyung Hee; Bae, Hyun Cheol; Kim, Seok-Ho; Yoo, Young Do; Seong, Seung-Yong; Kim, Tae Woo

    2009-01-29

    Dendritic cells (DCs) have become an important measure for the treatment of malignancies. Current DC preparations, however, generate short-lived DCs because they are subject to cell death from various apoptotic pressures. Antigen-specific CD8(+) cytotoxic T lymphocytes (CTLs) is one of the main obstacles to limit the DC-mediated immune priming since CTLs can recognize the target antigen expressing DCs as target cells and kill the DCs. CTLs secret perforin and serine protease granzymes during CTL killing. Perforin and serine protease granzymes induce the release of a number of mitochondrial pro-apoptotic factors, which are controlled by members of the BCL-2 family, such as BAK, BAX and BIM. FasL linking to Fas on DCs triggers the activation of caspase-8, which eventually leads to mitochondria-mediated apoptosis via truncation of BID. In this study, we tried to enhance the DC priming capacity by prolonging DC survival using anti-apoptotic siRNA targeting these key pro-apoptotic molecules in CTL killing. Human papillomavirus (HPV)-16 E7 antigen presenting DCs that were transfected with these anti-apoptotic siRNAs showed increased resistance to T cell-mediated death, leading to enhanced E7-specific CD8(+) T cell activation in vitro and in vivo. Among them, siRNA targeting BIM (siBIM) generated strongest E7-specific E7-specific CD8(+) T cell immunity. More importantly, vaccination with E7 presenting DCs transfected with siBIM was capable of generating a marked therapeutic effect in vaccinated mice. Our data indicate that ex vivo manipulation of DCs with siBIM may represent a plausible strategy for enhancing dendritic cell-based vaccine potency.

  6. Monocrystalline silicon photovoltaic luminescent solar concentrator with 4.2% power conversion efficiency.

    PubMed

    Desmet, L; Ras, A J M; de Boer, D K G; Debije, M G

    2012-08-01

    We report conversion efficiencies of experimental single and dual light guide luminescent solar concentrators. We have built several 5 cm × 5 cm and 10× cm × 10 cm luminescent solar concentrator (LSC) demonstrators consisting of c-Si photovoltaic cells attached to luminescent light guides of Lumogen F Red 305 dye and perylene perinone dye. The highest overall efficiency obtained was 4.2% on a 5 cm × 5 cm stacked dual light guide using both luminescent materials. To our knowledge, this is the highest reported experimentally determined efficiency for c-Si photovoltaic-based LSCs. Furthermore, we also produced a 5 cm × 5 cm LSC specimen based on an inorganic phosphor layer with an overall efficiency of 2.5%.

  7. An in-situ TEM study of the effects of 6 keV He ion irradiation on Si and SiO2

    NASA Astrophysics Data System (ADS)

    Donnelly, S. E.; Hinks, J. A.; Pawley, C. J.; Abrams, K. J.; van den Berg, J. A.

    2012-07-01

    Using the new MIAMI facility (Microscope and Ion Accelerators for Materials Investigations) at the University of Huddersfield, an in-situ TEM study has been carried out on the effects of helium implantation on a tri-layer system consisting of monocrystalline silicon (c-Si), silicon dioxide (SiO2) and polycrystalline silicon (poly-Si). Co-irradiation of these three components enabled direct comparisons to be made of differences between the response of c-Si and poly-Si to the He irradiation; in particular, differences in the development of interstitial clusters and bubbles and eventual amorphisation. For implantations to high fluences (>1017 ions/cm2), very significant levels of porosity were observed to build up in the Si, leading to changes in the width of the Si layers of up to 29% Although no helium bubbles were formed in the SiO2 layer, a very significant dimensional change (but in this case an observed shrinkage) also occurred in this material. Finally, room temperature amorphisation of the Si was observed at high fluences, beginning at somewhat lower fluences in the poly-Si than in the c-Si. A brief discussion of the origins of these effects is presented.

  8. Crystal structure of laser-induced subsurface modifications in Si

    SciTech Connect

    Verburg, P. C.; Smillie, L. A.; Römer, G. R. B. E.; Haberl, B.; Bradby, J. E.; Williams, J. S.; Huis in ’t Veld, A. J.

    2015-06-04

    Laser-induced subsurface modification of dielectric materials is a well-known technology. Applications include the production of optical components and selective etching. In addition to dielectric materials, the subsurface modification technology can be applied to silicon, by employing near to mid-infrared radiation. An application of subsurface modifications in silicon is laser-induced subsurface separation, which is a method to separate wafers into individual dies. Other applications for which proofs of concept exist are the formation of waveguides and resistivity tuning. However, limited knowledge is available about the crystal structure of subsurface modifications in silicon. In this paper, we investigate the geometry and crystal structure of laser-induced subsurface modifications in monocrystalline silicon wafers. Finally, in addition to the generation of lattice defects, we found that transformations to amorphous silicon and Si-iii/Si-xii occur as a result of the laser irradiation.

  9. Crystal structure of laser-induced subsurface modifications in Si

    DOE PAGES

    Verburg, P. C.; Smillie, L. A.; Römer, G. R. B. E.; ...

    2015-06-04

    Laser-induced subsurface modification of dielectric materials is a well-known technology. Applications include the production of optical components and selective etching. In addition to dielectric materials, the subsurface modification technology can be applied to silicon, by employing near to mid-infrared radiation. An application of subsurface modifications in silicon is laser-induced subsurface separation, which is a method to separate wafers into individual dies. Other applications for which proofs of concept exist are the formation of waveguides and resistivity tuning. However, limited knowledge is available about the crystal structure of subsurface modifications in silicon. In this paper, we investigate the geometry and crystalmore » structure of laser-induced subsurface modifications in monocrystalline silicon wafers. Finally, in addition to the generation of lattice defects, we found that transformations to amorphous silicon and Si-iii/Si-xii occur as a result of the laser irradiation.« less

  10. Folate-targeted star-shaped cationic copolymer co-delivering docetaxel and MMP-9 siRNA for nasopharyngeal carcinoma therapy

    PubMed Central

    Liu, Tao; Wu, Xidong; Wang, Yigang; Zhang, Tao; Wu, Ting; Liu, Fang; Wang, Wansong; Jiang, Gang; Xie, Minqiang

    2016-01-01

    The co-delivery of drug and gene has become the primary strategy in cancer therapy. Based on our previous work, to co-deliver docetaxel (DOC) and MMP-9 siRNA more efficiently for HNE-1 nasopharyngeal carcinoma therapy, a folate-modified star-shaped copolymer (FA-CD-PLLD) consisting of β-cyclodextrin (CD) and poly(L-lysine) dendron (PLLD) was synthesized, and then used for DOC and MMP-9 co-delivery. Different from commonly used amphiphilic copolymers micelles, the obtained CD derivative could be used directly for the combinatorial delivery of nucleic acid and hydrophobic DOC without a complicated micellization process. In vitro and in vivo assays are carried out to confirm the effectiveness of the target strategy and combined treatment. It was found that the conjugation of CD-PLLD with FA could enhance the DOC/MMP-9 delivery effect obviously, inducing a more significant apoptosis and decreasing invasive capacity of HEN-1 cells. In vivo assays showed that FA-CD-PLLD/DOC/MMP-9 could inhibit HNE-1 tumor growth and decrease PCNA expression effectively, indicating a promising strategy for nasopharyngeal carcinoma therapy. Moreover, the in vivo distribution of DOC and MMP-9, blood compatibility and toxicity are also explored. PMID:27259274

  11. Correlating animal and human phase Ia/Ib clinical data with CALAA-01, a targeted, polymer-based nanoparticle containing siRNA

    PubMed Central

    Zuckerman, Jonathan E.; Gritli, Ismael; Tolcher, Anthony; Heidel, Jeremy D.; Lim, Dean; Morgan, Robert; Chmielowski, Bartosz; Ribas, Antoni; Davis, Mark E.; Yen, Yun

    2014-01-01

    Nanoparticle-based experimental therapeutics are currently being investigated in numerous human clinical trials. CALAA-01 is a targeted, polymer-based nanoparticle containing small interfering RNA (siRNA) and, to our knowledge, was the first RNA interference (RNAi)–based, experimental therapeutic to be administered to cancer patients. Here, we report the results from the initial phase I clinical trial where 24 patients with different cancers were treated with CALAA-01 and compare those results to data obtained from multispecies animal studies to provide a detailed example of translating this class of nanoparticles from animals to humans. The pharmacokinetics of CALAA-01 in mice, rats, monkeys, and humans show fast elimination and reveal that the maximum concentration obtained in the blood after i.v. administration correlates with body weight across all species. The safety profile of CALAA-01 in animals is similarly obtained in humans except that animal kidney toxicities are not observed in humans; this could be due to the use of a predosing hydration protocol used in the clinic. Taken in total, the animal models do appear to predict the behavior of CALAA-01 in humans. PMID:25049380

  12. Folic acid-conjugated LaF3:Yb,Tm@SiO2 nanoprobes for targeting dual-modality imaging of upconversion luminescence and X-ray computed tomography.

    PubMed

    Ma, Jiebing; Huang, Peng; He, Meng; Pan, Liyuan; Zhou, Zhijun; Feng, Lili; Gao, Guo; Cui, Daxiang

    2012-12-06

    Development of multimodal contrast agents for in vivo simultaneous multimodality imaging is an emerging interdiscipline that is paving the avenue toward the goal of personalized medicine. Herein, folic acid-conjugated silica-modified LaF(3):Yb,Tm upconversion nanoparticles (UCNPs@SiO(2)-FA) with high La content in a single particle were strategically designed and prepared for simultaneously targeting dual-modality imaging of upconversion luminescence (UCL) and X-ray computed tomography (CT). LaF(3) UCNPs were synthesized by a novel oleic acid (OA)/ionic liquid (IL) two-phase system. Afterward, a folic acid molecule was covalently anchored on the surface of UCNPs with a silane coupling agent. The UCNPs@SiO(2)-FA exhibits good stability, water dispersibility and solubility, low cytotoxicity, good biocompatibility, highly selective targeting, excellent X-ray attenuation, and UCL emission under excitation at 980 nm. In vivo UCL and CT images of mice show the UCNPs@SiO(2)-FA can be used in targeting dual-modality imaging. These results suggest that the as-prepared nanoprobe is a good candidate with excellent imaging and targeting ability for targeting dual-modality imaging of UCL and CT.

  13. 5'-Triphosphate siRNA targeting MDR1 reverses multi-drug resistance and activates RIG-I-induced immune-stimulatory and apoptotic effects against human myeloid leukaemia cells.

    PubMed

    Li, Dengzhe; Gale, Robert Peter; Liu, Yanfeng; Lei, Baoxia; Wang, Yuan; Diao, Dongmei; Zhang, Mei

    2017-07-01

    Multi-drug resistance (MDR), immune suppression and decreased apoptosis are important causes of therapy-failure in leukaemia. Short interfering RNAs (siRNAs) down-regulate gene transcription, have sequence-independent immune-stimulatory effects and synergize with other anti-cancer therapies in some experimental models. We designed a siRNA targeting MDR1 with 5'-triphosphate ends (3p-siRNA-MDR1). Treatment of leukaemia cells with 3p-siRNA-MDR1 down-regulated MDR1 expression, reduced-drug resistance and induced immune and pro-apoptotic effects in drug-resistant HL-60/Adr and K562/Adr human leukaemia cell lines. We show mechanisms-of-action of these effects involve alterations in the anti-viral cytosolic retinoic acid-inducible protein-I (RIG-I; encoded by RIG-I or DDX58) mediated type-I interferon signal induction, interferon-gamma-inducible protein 10 (IP-10; encoded by IP10 or CXCL10) secretion, major histocompatibility complex-I expression (MHC-I) and caspase-mediated cell apoptosis. 3p-siRNA-MDR1 transfection also enhanced the anti-leukaemia efficacy of doxorubicin. These data suggest a possible synergistic role for 3p-siRNA-MDR1 in anti-leukaemia therapy. Copyright © 2017 Elsevier Ltd. All rights reserved.

  14. Behavior of oxygen doped SiC thin films: An x-ray photoelectron spectroscopy study

    NASA Astrophysics Data System (ADS)

    Avila, A.; Montero, I.; Galán, L.; Ripalda, J. M.; Levy, R.

    2001-01-01

    Thin silicon carbide films have been deposited by chemical vapor deposition on p-type (100) silicon substrates. The composition and bonds formed in these films have been analyzed by x-ray photoelectron spectroscopy (XPS) and infrared spectroscopy. The native surface oxide on the silicon carbide surface induced by air exposure has also been studied. Several phases are detected in the near-surface region: elemental Si, Si oxides (mainly SiO2), Si carbide (SiC) and Si oxicarbides (SiOxCy). Quantitative XPS analysis results indicate that, for atomic oxygen fractions <0.15, the Si-C phases are dominant in the films. Above this value no silicon oxicarbide is observed, but a multiphase material formed by elemental Si, Si oxides and Si carbides is observed. In spite of the film being a complex phase mixture, a simple relationship is found between the overall carbon and oxygen compositions. The carbon atomic fraction in the film decreases quasilinearly as the oxygen content increases, with a slope of about -1. An overall composition of SiOxC3-x in the 0.5monocrystalline silicon is made.

  15. Competitive diffusion of gold and copper atoms in Cu/Au/Si and Au/Cu/Si annealed systems

    NASA Astrophysics Data System (ADS)

    Benazzouz, C.; Benouattas, N.; Bouabellou, A.

    2005-04-01

    Thermal evaporation is used to deposit Au/Cu and Cu/Au bilayers on (1 1 1) monocrystalline silicon substrates. The layers composition and surface morphology are performed as function of annealing temperature at 200 and 400 °C using X-ray diffraction, Rutherford backscattering spectroscopy and scanning electron microscopy. Independently to the sequence of copper and gold deposition, Cu3Si and Cu4Si copper silicides result to the reaction and interdiffusion at the different interfaces. The 1000 and 1200 Å thickness of gold and copper diffusion barriers are insufficient to prevent the diffusion of copper and gold atoms respectively, after an annealing of only 200 °C. The fitting of concentration profiles of RBS spectra by using the RUMP software revealed a growth of silicides layers not uniform laterally and in depth.

  16. High-performance LWIR microbolometer with Si/SiGe quantum well thermistor and wafer level packaging

    NASA Astrophysics Data System (ADS)

    Roer, Audun; Lapadatu, Adriana; Wolla, Erik; Kittilsland, Gjermund

    2013-06-01

    An uncooled microbolometer with peak responsivity in the long wave infrared region of the electromagnetic radiation is developed at Sensonor AS. It is a 384 x 288 focal plane array with a pixel pitch of 25µm, based on monocrystalline Si/SiGe quantum wells as IR sensitive material. The high sensitivity (TCR) and low 1/f-noise are the main performance characteristics of the product. The frame rate is maximum 60Hz and the output interface is digital (LVDS). The quantum well thermistor material is transferred to the read-out integrated circuit (ROIC) by direct wafer bonding. The ROIC wafer containing the released pixels is bonded in vacuum with a silicon cap wafer, providing hermetic encapsulation at low cost. The resulting wafer stack is mounted in a standard ceramic package. In this paper the architecture of the pixels and the ROIC, the wafer packaging and the electro-optical measurement results are presented.

  17. Novel targeting of PEGylated liposomes for codelivery of TGF-β1 siRNA and four antitubercular drugs to human macrophages for the treatment of mycobacterial infection: a quantitative proteomic study

    PubMed Central

    Niu, Ning-Kui; Yin, Juan-Juan; Yang, Yin-Xue; Wang, Zi-Li; Zhou, Zhi-Wei; He, Zhi-Xu; Chen, Xiao-Wu; Zhang, Xueji; Duan, Wei; Yang, Tianxin; Zhou, Shu-Feng

    2015-01-01

    Tuberculosis (TB) is still a major public health issue in developing countries, and its chemotherapy is compromised by poor drug compliance and severe side effects. This study aimed to synthesize and characterize new multimodal PEGylated liposomes encapsulated with clinically commonly used anti-TB drugs with linkage to small interfering RNA (siRNA) against transforming growth factor-β1 (TGF-β1). The novel NP-siRNA liposomes could target THP-1-derived human macrophages that were the host cells of mycobacterium infection. The biological effects of the NP-siRNA liposomes were evaluated on cell cycle distribution, apoptosis, autophagy, and the gene silencing efficiency of TGF-β1 siRNA in human macrophages. We also explored the proteomic responses to the newly synthesized NP-siRNA liposomes using the stable isotope labeling with amino acids in cell culture approach. The results showed that the multifunctional PEGylated liposomes were successfully synthesized and chemically characterized with a mean size of 265.1 nm. The novel NP-siRNA liposomes functionalized with the anti-TB drugs and TGF-β1 siRNA were endocytosed efficiently by human macrophages as visualized by transmission electron microscopy and scanning electron microscopy. Furthermore, the liposomes showed a low cytotoxicity toward human macrophages. There was no significant effect on cell cycle distribution and apoptosis in THP-1-derived macrophages after drug exposure at concentrations ranging from 2.5 to 62.5 μg/mL. Notably, there was a 6.4-fold increase in the autophagy of human macrophages when treated with the NP-siRNA liposomes at 62.5 μg/mL. In addition, the TGF-β1 and nuclear factor-κB expression levels were downregulated by the NP-siRNA liposomes in THP-1-derived macrophages. The Ingenuity Pathway Analysis data showed that there were over 40 signaling pathways involved in the proteomic responses to NP-siRNA liposome exposure in human macrophages, with 160 proteins mapped. The top five canonical

  18. CVD-Based Valence-Mending Passivation for Crystalline-Si Solar Cells

    SciTech Connect

    Tao, Meng

    2015-03-01

    The objective of this project is to investigate a new surface passivation technique, valence-mending passivation, for its applications in crystalline-Si solar cells to achieve significant efficiency improvement and cost reduction. As the enabling technique, the project includes the development of chemical vapor deposition recipes to passivate textured Si(100) and multicrystalline-Si surfaces by sulfur and the characterization of the passivated Si surfaces, including thermal stability, Schottky barrier height, contact resistance and surface recombination. One important application is to replace the Ag finger electrode in Si cells with Al to reduce cost, by ~$0.1/Wp, and allow terawatt-scale deployment of crystalline-Si solar cells. These all-Al Si cells require a low-temperature metallization process for the Al electrode, to be compatible with valence-mending passivation and to prevent Al diffusion into n-type Si. Another application is to explore valence-mending passivation of grain boundaries in multicrystalline Si by diffusing sulfur into grain boundaries, to reduce the efficiency gas between monocrystalline-Si solar cells and multicrystalline-Si cells. The major accomplishments of this project include: 1) Demonstration of chemical vapor deposition processes for valence-mending passivation of both monocrystalline Si(100) and multicrystalline Si surfaces. Record Schottky barriers have been demonstrated, with the new record-low barrier of less than 0.08 eV between Al and sulfur-passivated n-type Si(100) and the new record-high barrier of 1.14 eV between Al and sulfur-passivated p-type Si(100). On the textured p-type monocrystalline Si(100) surface, the highest barrier with Al is 0.85 eV by valence-mending passivation. 2) Demonstration of a low-temperature metallization process for Al in crystalline-Si solar cells. The new metallization process is based on electroplating of Al in a room-temperature ionic liquid. The resistivity of the electroplated Al is ~7×10–6

  19. Nonlinear phenomenon in monocrystalline silicon based PV module for low power system: Lead acid battery for low energy storage

    NASA Astrophysics Data System (ADS)

    El Amrani, A.; El Amraoui, M.; El Abbassi, A.; Messaoudi, C.

    2014-11-01

    In the present work, we report the indoor photo-electrical measurements of monocrystalline silicon based photovoltaic (PV) module associated with 4 Ah lead acid battery as a storage unit for low power PV system applications. Concerning the PV module, our measurements show, at low illumination regime, that the short circuit current ISC increases linearly with the illumination power levels. Moreover, for high illumination levels, the mechanism of bimolecular recombination and space charge limitation may be intensified and hence the short current of the PV module ISCMod depends sublinearly on the incident optical power; the behavior is nonlinear. For the open circuit voltage of the PV module VOCMod measurements, a linear variation of the VOCMod versus the short circuit current in semi-logarithmic scale has been noticed. The diode ideality factor n and diode saturation current Is have been investigated; the values of n and Is are approximately of 1.3 and 10-9 A, respectively. In addition, we have shown, for different discharging-charging currents rates (i.e. 0.35 A, 0.2 A and 0.04 A), that the battery voltage decreases with discharging time as well as discharging battery capacity, and on the other hand it increases with the charging time and will rise up until it maximized value. The initial result shows the possibility to use such lead acid battery for low power PV system, which is generally designed for the motorcycle battery.

  20. Halide perovskite solar cells using monocrystalline TiO2 nanorod arrays as electron transport layers: impact of nanorod morphology

    NASA Astrophysics Data System (ADS)

    Thakur, Ujwal Kumar; Askar, Abdelrahman M.; Kisslinger, Ryan; Wiltshire, Benjamin D.; Kar, Piyush; Shankar, Karthik

    2017-07-01

    This is the first report of a 17.6% champion efficiency solar cell architecture comprising monocrystalline TiO2 nanorods (TNRs) coupled with perovskite, and formed using facile solution processing without non-routine surface conditioning. Vertically oriented TNR ensembles are desirable as electron transporting layers (ETLs) in halide perovskite solar cells (HPSCs) because of potential advantages such as vectorial electron percolation pathways to balance the longer hole diffusion lengths in certain halide perovskite semiconductors, ease of incorporating nanophotonic enhancements, and optimization between a high contact surface area for charge transfer (good) versus high interfacial recombination (bad). These advantages arise from the tunable morphology of hydrothermally grown rutile TNRs, which is a strong function of the growth conditions. Fluorescence lifetime imaging microscopy of the HPSCs demonstrated a stronger quenching of the perovskite PL when using TNRs as compared to mesoporous/compact TiO2 thin films. Due to increased interfacial contact area between the ETL and perovskite with easier pore filling, charge separation efficiency is dramatically enhanced. Additionally, solid-state impedance spectroscopy results strongly suggested the suppression of interfacial charge recombination between TNRs and perovskite layer, compared to other ETLs. The optimal ETL morphology in this study was found to consist of an array of TNRs ∼300 nm in length and ∼40 nm in width. This work highlights the potential of TNR ETLs to achieve high performance solution-processed HPSCs.

  1. Connections between morphological and mechanical evolution during galvanic corrosion of micromachined polycrystalline and monocrystalline silicon

    NASA Astrophysics Data System (ADS)

    Miller, David C.; Boyce, Brad L.; Kotula, Paul G.; Stoldt, Conrad R.

    2008-06-01

    Many microsystems fabrication technologies currently employ a metallic overlayer, such as gold, in electrical contact with silicon structural layers. During postprocessing in hydrofluoric-based acid solutions, a galvanic cell is created between the silicon and the metallic layer. Micromachined tensile specimens reveal that such etching in the presence of a galvanic cell can cause a catastrophic reduction in the tensile strength and apparent modulus of silicon. Detailed failure analysis was also used to compare fractured corroded Si to otherwise identical reference specimens via surface based (electron and scanning probe) microscopy as well as cross-section based structural- and composition-characterization techniques. For both polycrystalline and single-crystal silicon, galvanic corrosion can result in a thick corroded surface layer created via porous silicon formation, and/or generalized material removal depending on the etch chemistry and conditions. Under certain etching conditions, the porous silicon formation process results in cavity formation as well as preferential grain-boundary attack leading to intergranular fracture. The nature and severity of corrosion damage are shown to be influenced by the surface wetting characteristics of the etch chemistry, with poor wetting resulting in localized attack facilitated by the microstructure and good wetting resulting in generalized attack. The measured stiffness of the tensile specimens can be used to determine the effective modulus and porosity of the corroded surface layer. Extending beyond previous investigations, the present work examines the quantitative connection between the choice of chemical etchant, the corresponding damage morphology, and the resulting degradation in strength and apparent modulus. The present work also uniquely identifies important differences in polycrystalline and single-crystal Si based on their disparate damage evolution and related mechanical performance.

  2. Development of Targeted Recombinant Polymers that can deliver siRNA to the Cytoplasm and Plasmid DNA to the Cell Nucleus

    PubMed Central

    Canine, Brenda F.; Wang, Yuhua; Ouyang, Wenyun; Hatefi, Arash

    2011-01-01

    One of the major limitations to effective siRNA delivery is the lack of a siRNA-specific delivery system. Currently, the same delivery systems that are used for plasmid DNA (pDNA) delivery to the cell nucleus are used for siRNA delivery to the cytoplasm. To fill this gap, the objective of this study was to design a biopolymer that can be programmed via its amino acid sequence to deliver siRNA specifically to cytoplasm. For pDNA delivery, a nuclear localization signal (NLS) was added to the biopolymer structure to facilitate active translocation of the genetic material towards nucleus. The biopolymers were complexed with pEGFP and GFP-siRNA and used to transfect SKOV-3 (HER2+) cells. The intracellular trafficking of the nanoparticles was also monitored in real-time and live cells. The results demonstrated that the biopolymer with NLS is a suitable carrier for pDNA delivery but not siRNA delivery. Conversely, the biopolymer without NLS was suitable for siRNA delivery to the cytoplasm but not pDNA to the cell nucleus. The potential use of the designed biopolymer for combination therapy of cancer cells with gene (thymidine kinase) and siRNA (BCL2) was also examined in SKOV-3 cancer cells. PMID:21192992

  3. Development of targeted recombinant polymers that can deliver siRNA to the cytoplasm and plasmid DNA to the cell nucleus.

    PubMed

    Canine, Brenda F; Wang, Yuhua; Ouyang, Wenyun; Hatefi, Arash

    2011-04-10

    One of the major limitations to effective siRNA delivery is the lack of a siRNA-specific delivery system. Currently, the same delivery systems that are used for plasmid DNA (pDNA) delivery to the cell nucleus are used for siRNA delivery to the cytoplasm. To fill this gap, the objective of this study was to design a biopolymer that can be programmed via its amino acid sequence to deliver siRNA specifically to cytoplasm. For pDNA delivery, a nuclear localization signal (NLS) was added to the biopolymer structure to facilitate active translocation of the genetic material towards nucleus. The biopolymers were complexed with pEGFP and GFP-siRNA and used to transfect SKOV-3 (HER2+) cells. The intracellular trafficking of the nanoparticles was also monitored in real-time and live cells. The results demonstrated that the biopolymer with NLS is a suitable carrier for pDNA delivery but not siRNA delivery. Conversely, the biopolymer without NLS was suitable for siRNA delivery to the cytoplasm but not pDNA to the cell nucleus. The potential use of the designed biopolymer for combination therapy of cancer cells with gene (thymidine kinase) and siRNA (BCL2) was also examined in SKOV-3 cancer cells.

  4. Growth of Epitaxial SiSn Films with High Sn Content for IR Converters

    NASA Astrophysics Data System (ADS)

    Timofeev, V. A.; Nikiforov, A. I.; Kokhanenko, A. P.; Tuktamyshev, A. R.; Mashanov, V. I.; Loshkarev, I. D.; Novikov, V. A.

    2017-06-01

    Growth of SiSn compounds with a Sn content from 10 to 35% is studied. The morphology and surface structure of the SiSn layers are examined and the kinetic diagram of the morphological state of SiSn films is established in the temperature range of 150-450°C. During the growth of SiSn films from 150 to 300°C, oscillations of specular beam were observed. For the first time, periodic multilayer SiSn/Si structures with pseudomorphic monocrystalline SiSn layers with the Sn content from 10 to 25% are grown. The c(8×4) and (5×1) superstructures are identified during the growth of Si on the SiSn layer and the conditions are determined for the formation of the desired Si surface structure by controlling the growth temperature. From the diffraction reflection curves, the lattice parameter, the SiSn composition, and the period in the multilayer periodic structure are defined, which with high precision correspond to the specified values.

  5. Efficient siRNA delivery and tumor accumulation mediated by ionically cross-linked folic acid-poly(ethylene glycol)-chitosan oligosaccharide lactate nanoparticles: for the potential targeted ovarian cancer gene therapy.

    PubMed

    Li, Tony Shing Chau; Yawata, Toshio; Honke, Koichi

    2014-02-14

    For effective ovarian cancer gene therapy, systemic administrated tumor-targeting siRNA/folic acid-poly(ethylene glycol)-chitosan oligosaccharide lactate (FA-PEG-COL) nanoparticles is vital for delivery to cancer site(s). siRNA/FA-PEG-COL nanoparticles were prepared by ionic gelation for effective FA receptor-expressing ovarian cancer cells transfection and in vivo accumulation. The chemical structure of FA-PEG-COL conjugate was characterized by MALDI-TOF-MS, FT-IR and (1)H NMR. The average size of siRNA/FA-PEG-COL nanoparticles was approximately 200 nm, and the surface charge was +8.4 mV compared to +30.5 mV with siRNA/COL nanoparticles. FA-PEG-COL nanoparticles demonstrated superior compatibility with erythrocytes in terms of degree of aggregation and haemolytic activity and also effects on cell viability was lower when compared with COL nanoparticles. FA grafting significantly facilitated the uptake of nanoparticles via receptor mediated endocytosis as demonstrated by flow cytometry. The in vitro transfection and gene knockdown efficiency of HIF-1α were superior to COL nanoparticles (76-62%, respectively) and was comparable to Lipofectamine 2000 (79%) as demonstrated by RT-qPCR and Western blot. Gene knockdown at the molecular level translated into effective inhibition of proliferation in vitro. Accumulation efficiency of FA-PEG-COL nanoparticles was investigated in BALB/c mice bearing OVK18 #2 tumor xenograft using in vivo imaging. The active targeting FA-PEG-COL nanoparticles showed significantly greater accumulation than the passive targeting COL nanoparticles. Based on the results obtained, siRNA/FA-PEG-COL nanoparticles show much potential for effective ovarian cancer treatment via gene therapy.

  6. Study by simulation of the SnO2 and ZnO anti-reflection layers in n-SiC/p-SiC solar cells

    NASA Astrophysics Data System (ADS)

    Zerfaoui, Hana; Dib, Djalel; Rahmani, Mohamed; Benyelloul, Kamel; Mebarkia, Chafia

    2016-07-01

    Recently, Two technologies of the photovoltaic cells are present today namely the cells crystalline (polycrystalline and monocrystalline) and the cell thin layers. The development of the solar cells requires a technological change of materials used in their manufacturing. The thin layers are parts of these materials and which announced their effectiveness and growth of output of the solar cell. The aim of this paper article is to the study and simulation of photovoltaic cells containing SiC materials. This material is have important having a part in the development of renewable energies. Based on the SCAPS (a Solar Cell Capacitance Simulator) simulation, the obtained results are Vco, Jsc, FF and the output energy of conversion of a solar cell n-SiC/p-SiC with different materials for the anti-reflecting layer ZnO and SnO2.with the SCAPS (a Solar Cell Capacitance Simulator) computer code in one dimension, the results obtained after optimization.

  7. Boron diffusion in nanocrystalline 3C-SiC

    SciTech Connect

    Schnabel, Manuel; Weiss, Charlotte; Rachow, Thomas; Löper, Philipp; Janz, Stefan; Canino, Mariaconcetta; Summonte, Caterina; Mirabella, Salvo; Wilshaw, Peter R.

    2014-05-26

    The diffusion of boron in nanocrystalline silicon carbide (nc-SiC) films with a grain size of 4–7 nm is studied using a poly-Si boron source. Diffusion is found to be much faster than in monocrystalline SiC as it takes place within the grain boundary (GB) network. Drive-in temperatures of 900–1000°C are suitable for creating shallow boron profiles up to 100 nm deep, while 1100°C is sufficient to flood the 200 nm thick films with boron. From the resulting plateau at 1100 °C a boron segregation coefficient of 28 between nc-SiC and the Si substrate, as well as a GB boron solubility limit of 0.2 nm{sup −2} is determined. GB diffusion in the bulk of the films is Fickian and thermally activated with D{sub GB}(T)=(3.1−5.6)×10{sup 7}exp(−5.03±0.16  eV/k{sub B}T) cm{sup 2}s{sup −1}. The activation energy is interpreted in terms of a trapping mechanism at dangling bonds. Higher boron concentrations are present at the nc-SiC surface and are attributed to immobilized boron.

  8. Unveiling the Hybrid n-Si/PEDOT:PSS Interface.

    PubMed

    Jäckle, Sara; Liebhaber, Martin; Niederhausen, Jens; Büchele, Matthias; Félix, Roberto; Wilks, Regan G; Bär, Marcus; Lips, Klaus; Christiansen, Silke

    2016-04-06

    We investigated the buried interface between monocrystalline n-type silicon (n-Si) and the highly conductive polymer poly(3,4-ethylenedioxythiophene)-poly(styrenesulfonate) ( PSS), which is successfully applied as a hole selective contact in hybrid solar cells. We show that a post-treatment of the polymer films by immersion in a suitable solvent reduces the layer thickness by removal of excess material. We prove that this post-treatment does not affect the functionality of the hybrid solar cells. Through the thin layer we are probing the chemical structure at the n-Si/ PSS interface with synchrotron-based hard X-ray photoelectron spectroscopy (HAXPES). From the HAXPES data we conclude that the Si substrate of a freshly prepared hybrid solar cell is already oxidized immediately after preparation. Moreover, we show that even when storing the sample in inert gas such as, e.g., nitrogen the n-Si/SiOx/ PSS interface continues to further oxidize. Thus, without further surface treatment, an unstable Si suboxide will always be present at the hybrid interface.

  9. A small interfering RNA targeting vascular endothelial growth factor efficiently inhibits growth of VX2 cells and VX2 tumor model of hepatocellular carcinoma in rabbit by transarterial embolization-mediated siRNA delivery

    PubMed Central

    Zou, Yu; Guo, Chuan-Gen; Yang, Zheng-Gang; Sun, Jun-Hui; Zhang, Min-Ming; Fu, Cai-Yun

    2016-01-01

    -mediated VEGF-siRNA had no toxicity to the livers and kidneys of rabbits, and VEGF-siRNA had the ability to protect liver damage induced by tumor growth. Conclusion This is the first study to demonstrate that targeting VEGF via TAE-mediated siRNA delivery may become a powerful new option for effective treatment of hepatocellular carcinoma in the clinic. PMID:27069355

  10. Formation Monocrystalline Carbon Micro-and Nanostructures Under Femtosecond Laser Irradiation of graphite in Liquid Nitrogen

    NASA Astrophysics Data System (ADS)

    Khorkov, Kirill S.; Abramov, Dmitrii V.; Kochuev, Dmitriy A.; Arakelian, Sergey M.; Prokoshev, Valery G.

    The combination of high energy and ultra short duration of femtosecond laser pulses allow to reach in the area of impact the local conditions which can change the phase composition of irradiated material. Traditional methods of structural phase transformation of the graphite at high pressures do not provide the abrupt simultaneous cancellation of the applied pressure and temperature. As a result, some of the synthesized nanostructures and metastable forms of carbon are destroyed. The suggested method allows to eliminate this disadvantage. Femtosecond laser radiation provides ultrafast heating of the target material, and the use of liquid nitrogen dramatically accelerates the process of it cooling. The formation of new carbon micro- and nanostructures has been registered at experimental approbation of the proposed method. The check of elemental composition of the created crystals showed that they are formed solely of carbon. The experimental results show the possibility of creation of new (less studied) carbon forms with a variety of properties.

  11. Magnetically targeted delivery of DOX loaded Cu9S5@mSiO2@Fe3O4-PEG nanocomposites for combined MR imaging and chemo/photothermal synergistic therapy

    NASA Astrophysics Data System (ADS)

    Liu, Bei; Zhang, Xinyang; Li, Chunxia; He, Fei; Chen, Yinyin; Huang, Shanshan; Jin, Dayong; Yang, Piaoping; Cheng, Ziyong; Lin, Jun

    2016-06-01

    The combination of multi-theranostic modes in a controlled fashion has received tremendous attention for the construction of cooperative therapeutic systems in nanomedicine. Herein, we have synthesized a smart magnetically targeted nanocarrier system, Cu9S5@mSiO2@Fe3O4-PEG (labelled as CMF), which integrates NIR triggered photothermal therapy, pH/NIR-responsive chemotherapy and MR imaging into one nanoplatform to enhance the therapeutic efficacy. This new multifunctional paradigm has a uniform and monodisperse sesame ball-like structure by decorating tiny Fe3O4 nanoparticles on the surface of Cu9S5@mSiO2 before a further PEG modification to improve its hydrophilicity and biocompatibility. With doxorubicin (DOX) payload, the as-obtained CMF-DOX composites can simultaneously provide an intense heating effect and enhanced DOX release upon 980 nm NIR light exposure, achieving a combined chemo/photothermal therapy. Under the influence of an external magnetic field, the magnetically targeted synergistic therapeutic effect of CMF-DOX can lead to highly superior inhibition of animal H22 tumor in vivo when compared to any of the single approaches alone. The results revealed that this Cu9S5 based magnetically targeted chemo/photothermal synergistic nanocarrier system has great promise in future MR imaging assisted tumor targeted therapy of cancer.

  12. Construction of pH-responsive and up-conversion luminescent NaYF4:Yb3+/Er3+@SiO2@PMAA nanocomposite for colon targeted drug delivery

    PubMed Central

    Tian, Boshi; Liu, Shaohua; Lu, Wei; Jin, Lin; Li, Qingfeng; Shi, Yurong; Li, Chunyang; Wang, Zhenling; Du, Yaping

    2016-01-01

    Colon-targeted drug delivery system has attracted much interest because it can improve therapeutic efficacy and reduce the side effect in practical clinic. Herein, we constructed a multifunctional drug delivery system with colonic targeting and tracking by up-conversion (UC) luminescence based on core-shell structured NaYF4:Yb3+/Er3+@SiO2@PMAA nanocomposite. The resultant materials exhibited bright UC luminescence, pH-responsive property and excellent biocompatibility. The drug release behaviors in different pH environment were investigated using 5-aminosalicylic acid (5-ASA) as a model drug. The 5-ASA molecules release from NaYF4:Yb3+/Er3+@SiO2@PMAA nanocomposite exhibit a significant pH-responsive colon targeted property, i.e., a little amount of drug release in simulated gastric fluid (SGF, pH = 1.2) but a large amount of drug release in simulated colonic fluid (SCF, pH = 7.4) Moreover, the drug release process could be monitored by the change of UC emission intensity. These results implied that the multifunctional nanocomposite is a promising drug carrier for targeted release of 5-ASA in the colon. PMID:26891778

  13. Construction of pH-responsive and up-conversion luminescent NaYF4:Yb3+/Er3+@SiO2@PMAA nanocomposite for colon targeted drug delivery

    NASA Astrophysics Data System (ADS)

    Tian, Boshi; Liu, Shaohua; Lu, Wei; Jin, Lin; Li, Qingfeng; Shi, Yurong; Li, Chunyang; Wang, Zhenling; Du, Yaping

    2016-02-01

    Colon-targeted drug delivery system has attracted much interest because it can improve therapeutic efficacy and reduce the side effect in practical clinic. Herein, we constructed a multifunctional drug delivery system with colonic targeting and tracking by up-conversion (UC) luminescence based on core-shell structured NaYF4:Yb3+/Er3+@SiO2@PMAA nanocomposite. The resultant materials exhibited bright UC luminescence, pH-responsive property and excellent biocompatibility. The drug release behaviors in different pH environment were investigated using 5-aminosalicylic acid (5-ASA) as a model drug. The 5-ASA molecules release from NaYF4:Yb3+/Er3+@SiO2@PMAA nanocomposite exhibit a significant pH-responsive colon targeted property, i.e., a little amount of drug release in simulated gastric fluid (SGF, pH = 1.2) but a large amount of drug release in simulated colonic fluid (SCF, pH = 7.4) Moreover, the drug release process could be monitored by the change of UC emission intensity. These results implied that the multifunctional nanocomposite is a promising drug carrier for targeted release of 5-ASA in the colon.

  14. Theoretical and empirical studies of impurity incorporation into beta-SiC thin films during epitaxial growth

    NASA Astrophysics Data System (ADS)

    Kim, H. J.; Davis, R. F.

    1986-11-01

    A theoretical determination of the vapor species present, and their respective partial pressures, is made using the SOLGASMIX-PV program for the n-type and p-type dopants of N and P, and B, respectively, under conditions used to grow monocrystalline beta-SiC thin films via CVD. The model shows that Al and P behave ideally while B and N apparently interact with the C or Si in the SiC or fill normally unoccupied interstitial positions. The relationship between the carrier concentrations or the atomic concentrations and the partial pressure of the dopant source gases is linear and parallel. The more efficient n-type and p-type dopants of N and Al have been used to produce what is suggested to be the first p-n junction diode in a beta-SiC film.

  15. The effect of substrate type on SiC nanowire orientation.

    PubMed

    Attolini, Giovanni; Rossi, Francesca; Bosi, Matteo; Watts, Bernard Enrico; Salviati, Giancarlo

    2011-05-01

    beta-SiC nanowires were synthesized on different monocrystalline substrates: Si (001), Si (111), 3C-SiC (001), 4H-SiC (0001), 6H-SiC (0001). The SiC nanowire growth was carried out using a Chemical Vapor Deposition method, with silane and propane diluted in hydrogen (3%) as precursors. The deposition was performed at atmospheric pressure and at 1100 degrees C, after dewetting of the Ni catalyst, which had been previously evaporated onto the substrate, to induce 1D growth according to a VLS process. The crystal structure of the nanowires, as determined by X-ray diffraction and High Resolution Transmission Electron Microscopy, corresponds to 3C-SiC polytype growing along a (111) direction, irrespective of the substrate. The occurrence of (111) stacking faults was observed, partly reduced for samples grown on 3C-SiC substrate. The growth on (111) substrate allowed to achieve a good vertical alignment of the nanowires, as investigated by Scanning Electron Microscopy. High Angle Annular Dark Field imaging and Energy Dispersive X-Ray spectroscopy were performed to study the catalyst particle on top of the wires and showed the formation of a nickel-silicon alloy.

  16. Assessment of Nanobiotechnology-Targeted siRNA Designed to inhibit NF-kappaB Classical and Alternative Signaling in Breast Tumor Macrophages

    DTIC Science & Technology

    2013-07-01

    80:1183-96. 5 . Murdoch, C., et al., The role of myeloid cells in the promotion of tumour angiogenesis. Nat Rev Cancer, 2008. 8:618-31. 6. Pollard... Tumour -educated macrophages promote tumour progression and metastasis. Nature Reviews Cancer 4, 71-78 (2004). 5 . Condeelis, J. & Pollard, J.W...using siRNA to knockdown the IκBα classical inhibitor protein in these cells (Figure 5 ). When free siRNAs or empty Mn-NPs are incubated with BMDMs

  17. Dual-targeting and pH/redox-responsive multi-layered nanocomplexes for smart co-delivery of doxorubicin and siRNA.

    PubMed

    Han, Lu; Tang, Cui; Yin, Chunhua

    2015-08-01

    Multi-layered nanocomplexes (MLNs) were designed here to provide smart co-delivery of doxorubicin (DOX) and vascular endothelial growth factor (VEGF) siRNA. The electrostatically self-assembled MLNs were constructed by TAT peptide modified mesoporous silica nanoparticles (TAT-MSN) as the cationic core for DOX loading, poly(allylamine hydrochloride)-citraconic anhydride (PAH-Cit) as the anionic inner layer, and galactose-modified trimethyl chitosan-cysteine (GTC) conjugate as the cationic outer layer to encapsulate siRNA. Their strong stability at pH 7.4 and 6.5 protected siRNA from degradation in the blood and tumor microenvironment. Galactose ligands on the GTC outer layers effectively facilitated the internalization of MLNs through receptor-mediated endocytosis. Afterwards, the endosomal/lysosomal acidity (pH 5.0) triggered the charge reversal of PAH-Cit, thereby inducing the disassembly of MLNs and their escape to the cytosol. Cytoplasmic glutathione further accelerated siRNA release through cleaving disulfide bonds in GTC layers, leading to high silencing efficiencies. Meanwhile, the exposed DOX-loaded cores were transported into the nuclei by virtue of TAT peptide and exhibited sustained release thereafter. As a result, potent antitumor efficacies of MLNs were noted following intravenous injection at a low dose with no apparent toxicity detected. Therefore, MLNs served as an effective and safe vector to maximize synergistic effect of chemodrugs and therapeutic genes. Copyright © 2015 Elsevier Ltd. All rights reserved.

  18. Germ Cell-Specific Targeting of DICER or DGCR8 Reveals a Novel Role for Endo-siRNAs in the Progression of Mammalian Spermatogenesis and Male Fertility

    PubMed Central

    Zimmermann, Céline; Romero, Yannick; Warnefors, Maria; Bilican, Adem; Borel, Christelle; Smith, Lee B.; Kotaja, Noora; Kaessmann, Henrik; Nef, Serge

    2014-01-01

    Small non-coding RNAs act as critical regulators of gene expression and are essential for male germ cell development and spermatogenesis. Previously, we showed that germ cell-specific inactivation of Dicer1, an endonuclease essential for the biogenesis of micro-RNAs (miRNAs) and endogenous small interfering RNAs (endo-siRNAs), led to complete male infertility due to alterations in meiotic progression, increased spermatocyte apoptosis and defects in the maturation of spermatozoa. To dissect the distinct physiological roles of miRNAs and endo-siRNAs in spermatogenesis, we compared the testicular phenotype of mice with Dicer1 or Dgcr8 depletion in male germ cells. Dgcr8 mutant mice, which have a defective miRNA pathway while retaining an intact endo-siRNA pathway, were also infertile and displayed similar defects, although less severe, to Dicer1 mutant mice. These included cumulative defects in meiotic and haploid phases of spermatogenesis, resulting in oligo-, terato-, and azoospermia. In addition, we found by RNA sequencing of purified spermatocytes that inactivation of Dicer1 and the resulting absence of miRNAs affected the fine tuning of protein-coding gene expression by increasing low level gene expression. Overall, these results emphasize the essential role of miRNAs in the progression of spermatogenesis, but also indicate a role for endo-siRNAs in this process. PMID:25244517

  19. Adenovirus-mediated siRNA targeting TNF-α and overexpression of bone morphogenetic protein-2 promotes early osteoblast differentiation on a cell model of Ti particle-induced inflammatory response in vitro.

    PubMed

    Guo, H H; Yu, C C; Sun, S X; Ma, X J; Yang, X C; Sun, K N; Jin, Q H

    2013-10-01

    Wear particles are phagocytosed by macrophages and other inflammatory cells, resulting in cellular activation and release of proinflammatory factors, which cause periprosthetic osteolysis and subsequent aseptic loosening, the most common causes of total joint arthroplasty failure. During this pathological process, tumor necrosis factor-alpha (TNF-α) plays an important role in wear-particle-induced osteolysis. In this study, recombination adenovirus (Ad) vectors carrying both target genes [TNF-α small interfering RNA (TNF-α-siRNA) and bone morphogenetic protein 2 (BMP-2)] were synthesized and transfected into RAW264.7 macrophages and pro-osteoblastic MC3T3-E1 cells, respectively. The target gene BMP-2, expressed on pro-osteoblastic MC3T3-E1 cells and silenced by the TNF-α gene on cells, was treated with titanium (Ti) particles that were assessed by real-time PCR and Western blot. We showed that recombinant adenovirus (Ad-siTNFα-BMP-2) can induce osteoblast differentiation when treated with conditioned medium (CM) containing RAW264.7 macrophages challenged with a combination of Ti particles and Ad-siTNFα-BMP-2 (Ti-ad CM) assessed by alkaline phosphatase activity. The receptor activator of nuclear factor-κB ligand was downregulated in pro-osteoblastic MC3T3-E1 cells treated with Ti-ad CM in comparison with conditioned medium of RAW264.7 macrophages challenged with Ti particles (Ti CM). We suggest that Ad-siTNFα-BMP-2 induced osteoblast differentiation and inhibited osteoclastogenesis on a cell model of a Ti particle-induced inflammatory response, which may provide a novel approach for the treatment of periprosthetic osteolysis.

  20. Re-examination of siRNA specificity questions role of PICH and Tao1 in the spindle checkpoint and identifies Mad2 as a sensitive target for small RNAs.

    PubMed

    Hübner, Nadja C; Wang, Lily Hui-Ching; Kaulich, Manuel; Descombes, Patrick; Poser, Ina; Nigg, Erich A

    2010-04-01

    The DNA-dependent adenosine triphosphatase (ATPase) Plk1-interacting checkpoint helicase (PICH) has recently been implicated in spindle checkpoint (SAC) signaling (Baumann et al., Cell 128(1):101-114, 2007). Depletion of PICH by siRNA abolished the SAC and resulted in an apparently selective loss of Mad2 from kinetochores, suggesting a role for PICH in the regulation of the Mad1-Mad2 interaction. An apparent rescue of SAC functionality by overexpression of PICH in PICH-depleted cells initially seemed to confirm a role for PICH in the SAC. However, we have subsequently discovered that all PICH-directed siRNA oligonucleotides that abolish the SAC also reduce Mad2 mRNA and protein expression. This reduction is functionally significant, as PICH siRNA does not abolish SAC activity in a cell line that harbors a bacterial artificial chromosome driving the expression of murine Mad2. Moreover, we identified several siRNA duplexes that effectively deplete PICH but do not significantly affect SAC functionality or Mad2 abundance or localization. Finally, we discovered that the ability of overexpressed PICH to restore SAC activity in PICH-depleted cells depends on sequestration of the mitotic kinase Plk1 rather than ATPase activity of PICH, pointing to an underlying mechanism of "bypass suppression." In support of this view, depletion or inhibition of Plk1 also rescued SAC activity in cells harboring low levels of Mad2. This observation suggests that a reduction of Plk1 activity partially compensates for reduced Mad2 levels and argues that Plk1 normally reduces the strength of SAC signaling. Collectively, our results question the role of PICH in the SAC and instead identify Mad2 as a sensitive off target for small RNA duplexes. In support of the latter conclusion, our evidence suggests that an off-target effect on Mad2 may also contribute to explain the apparent role of the Tao1 kinase in SAC signaling.

  1. Systemic siRNA Delivery with a Dual pH-Responsive and Tumor-targeted Nanovector for Inhibiting Tumor Growth and Spontaneous Metastasis in Orthotopic Murine Model of Breast Carcinoma

    PubMed Central

    Fan, Bo; Kang, Lin; Chen, Liqing; Sun, Ping; Jin, Mingji; Wang, Qiming; Bae, You Han; Huang, Wei; Gao, Zhonggao

    2017-01-01

    Phenylboronic acid (PBA)-mediated tumor targeting nanovector is an attractive strategy for enhancing siRNA delivery and treatment of metastatic cancers. However, its nonspecific binding with various biological membranes containing cis-diol moieties restricts its potential application by systematic administration. Herein, we constructed a novel pH-activated “sheddable” PEG-coated nanoparticle for effective treatment of primary tumors and metastases, which was based on the conjugation of catechol group modified poly(ethylene glycol) (PEG-Cat) and PBA-terminated polyethylenimine (PEI-PBA) via the borate ester formed between PBA and Cat. By virtue of the pH-dependent stability of borate ester in an aqueous medium, the PEG-shell could “shield” the PBA ligand in systemic circulation to reduce its “off-target effect”, while PEG was detached at tumor extracellular pH (~6.5) to expose intact PBA moiety. Simultaneously, the PBA ligand could bind with overexpressed sialic acid residues on cancer cells, giving rise to enhanced cellular internalization. In addition, the PBA moieties could also couple with each 3'-end ribose of double-stranded siRNA. siRNAs were used as both a payload and a pH-responsive intermolecular cross-linker, and thereby acquired sufficient stability during circulating in blood and a rapidly triggered release in response to acidic endosomal/lysosomal pH-stimuli. As a result, this dual pH-sensitive nanoparticle showed enhanced siRNA uptake, gene silencing efficacy and anti-metastatic effects in vitro. Furthermore, in vivo studies demonstrated that PBA-based nanoparticles effectively accumulated in tumor and inhibited tumor growth and metastasis in 4T1 orthotopic mammary tumor model after intravenous administration. PMID:28042340

  2. Performance Comparison of Stion CIGS Modules to Baseline Monocrystalline Modules at the New Mexico Florida and Vermont Regional Test Centers: January 2015-December 2016.

    SciTech Connect

    Lave, Matthew Samuel; Stein, Joshua; Burnham, Laurie

    2017-01-01

    This report provides performance data and analysis for two Stion copper indium gallium selenide (CIGS) module types, one framed, the other frameless, and installed at the New Mexico, Florida and Vermont RTCs. Sandia looked at data from both module types and compared the latter with data from an adjacent monocrystalline baseline array at each RTC. The results indicate that the Stion modules are slightly outperforming their rated power, with efficiency values above 100% of rated power, at 25degC cell temperatures. In addition, Sandia sees no significant performance differences between module types, which is expected because the modules differ only in their framing. In contrast to the baseline systems, the Stion strings showed increasing efficiency with increasing irradiance, with the greatest increase between zero and 400 Wm -2 but still noticeable increases at 1000 Wm -2 . Although baseline data availability in Vermont was spotty and therefore comparative trends are difficult to discern, the Stion modules there may offer snow- shedding advantages over monocrystalline-silicon modules but these findings are preliminary.

  3. Assessment of Nanobiotechnology-Targeted siRNA Designed to Inhibit NF-KappaB Classical and Alternative Signaling in Breast Tumor Macrophages

    DTIC Science & Technology

    2014-07-01

    cardiovascular disease . To maximize... disease  with  the  poorest  clinical  outcomes.         We   tested   the   following  hypothesis:   siRNA-­‐mediated...in  Benign  and  Malignant   Disease .  Vanderbilt  University  Department   of  Medicine’s  Dinner  and  Data

  4. Target rapidity baryon distributions in {sup 28}Si + {sup 197}Au and {sup 197}Au + {sup 197}Au collisions at 14.6 and 11.7 A{center_dot}GeV/c

    SciTech Connect

    Sangster, T.C.; Costales, J.B.; Namboodiri, M.N.; E802 Collaboration

    1993-02-25

    Proton and deuteron kinetic energy spectra have been measured at target rapidities for both minimum bias and central collisions of 14.6 A{center_dot}GeV/c {sup 28}Si and 11.7 A{center_dot}GeV/c {sup 197}Au beams with a {sup 197}Au target. The spectra were measured from a low energy threshold of approximately E{sub kin}=35 MeV to well over 200 MeV for laboratory angles of 50{degree} to 130{degree} ({vert_bar}{eta}{vert_bar} {le}0.76). The acceptance-corrected spectra have been fit over a limited range of kinetic energies using a Boltzmann distribution. The integrated yields and the inverse slope parameters are presented as a function of centrality for the {sup 28}Si + {sup 197}Au reaction and as a function of trigger for the {sup 197}Au + {sup 197}Au reaction. These quantities are also compared with the proton spectra generated using both the ARC and RQMD codes.

  5. Assembly of Multifunctional Phi29 pRNA Nanoparticles for Specific Delivery of SiRNA and other Therapeutics to Targeted Cells

    PubMed Central

    Shu, Yi; Cinier, Mathieu; Shu, Dan; Guo, Peixuan

    2011-01-01

    Recent advances in RNA nanotechnology have led to the emergence of a new field and brought vitality to the area of therapeutics (Guo P, The Emerging Field of RNA Nanotechnology, Nature Nanotechnology, 2010). Due to the complementary nature of the four nucleotides and its special catalytic activity, RNA can be manipulated with simplicity characteristic of DNA, while possessing versatile structure and diverse function similar to proteins. Loops and tertiary architecture serve as mounting dovetails or wedges to eliminate external linking dowels. Unique features in transcription, termination, self-assembly, self-processing, and acid-resistance enable in vivo production of nanoparticles harboring aptamer, siRNA, ribozyme, riboswitch, or other regulators for therapy, detection, regulation, and intracellular computation. The unique property of noncanonical base-pairing and stacking enables RNA to fold into well-defined structures for constructing nanoparticles with special functionalities. Bacteriophage phi29 DNA packaging motor is geared by a ring consisting of six packaging RNA (pRNA) molecules. pRNA is able to form a multimeric complex via the interaction of two reengineered interlocking loops. This unique feature makes it an ideal polyvalent vehicle for nanomachine fabrication, pathogen detection, and delivery of siRNA or other therapeutics. This review describes methods in using pRNA as a building block for the construction of RNA dimers, trimers and hexamers as nanoparticles in medical applications. Methods for industrial-scale production of large and stable RNA nanoparticles will be introduced. The unique favorable PK (pharmokinetics) profile with a half life (T1/2) of 5–10 hours comparing to 0.25 of conventional 2′-F siRNA, and advantageous in vivo features such as non-toxicity, non-induction of interferons or non-stimulating of cytokine response in animals will also be reviewed. PMID:21320601

  6. Assembly of multifunctional phi29 pRNA nanoparticles for specific delivery of siRNA and other therapeutics to targeted cells.

    PubMed

    Shu, Yi; Cinier, Mathieu; Shu, Dan; Guo, Peixuan

    2011-06-01

    Recent advances in RNA nanotechnology have led to the emergence of a new field and brought vitality to the area of therapeutics [P. Guo, The emerging field of RNA nanotechnology, Nat. Nanotechnol., 2010]. Due to the complementary nature of the four nucleotides and its special catalytic activity, RNA can be manipulated with simplicity characteristic of DNA, while possessing versatile structure and diverse function similar to proteins. Loops and tertiary architecture serve as mounting dovetails or wedges to eliminate external linking dowels. Unique features in transcription, termination, self-assembly, self-processing, and acid-resistance enable in vivo production of nanoparticles harboring aptamer, siRNA, ribozyme, riboswitch, or other regulators for therapy, detection, regulation, and intracellular computation. The unique property of noncanonical base-pairing and stacking enables RNA to fold into well-defined structures for constructing nanoparticles with special functionalities. Bacteriophage phi29 DNA packaging motor is geared by a ring consisting of six packaging RNA (pRNA) molecules. pRNA is able to form a multimeric complex via the interaction of two reengineered interlocking loops. This unique feature makes it an ideal polyvalent vehicle for nanomachine fabrication, pathogen detection, and delivery of siRNA or other therapeutics. This review describes methods in using pRNA as a building block for the construction of RNA dimers, trimers, and hexamers as nanoparticles in medical applications. Methods for industrial-scale production of large and stable RNA nanoparticles will be introduced. The unique favorable PK (pharmacokinetics) profile with a half life (T(1/2)) of 5-10h comparing to 0.25 of conventional 2'-F siRNA, and advantageous in vivo features such as non-toxicity, non-induction of interferons or non-stimulating of cytokine response in animals will also be reviewed.

  7. Targeted knockout of TNF-α by injection of lentivirus-mediated siRNA into the subacromial bursa for the treatment of subacromial bursitis in rats.

    PubMed

    Wang, Yi; Li, Quan; Wei, Xianzhao; Xu, Jie; Chen, Qi; Song, Shuang; Lu, Zhe; Wang, Zimin

    2015-09-01

    Subacromial bursitis (SAB) is the major source of pain in rotator cuff disease. Although multiple investigations have provided support for the role of inflammatory cytokines in SAB, few have focussed on the use these cytokines in the treatment of SAB. The aim of the present study was to observe the therapeutic efficacy of lentivirus‑mediated RNA interference (RNAi) on carrageenan‑induced SAB by injecting lentivirus‑tumor necrosis factor (TNF)‑α‑RNAi expressing TNF‑α small interfering (si)RNA. Using screened siRNA segments, an siRNA was designed. A lentivirus vector expressing siRNA was established and packed as lentivirus particles. A lentivirus that expressed the negative sequence was used as a lentivirus‑negative control (NC). The carrageenan‑induced SAB model was established in 32 male Sprague‑Dawley rats. The modeled rats were randomly assigned to four groups: Lentivirus‑RNAi treatment group, lentivirus‑NC group, SAB group and phosphate‑buffered saline (PBS) blank control group. The lentivirus was injected (1x10(7) transducing units) into the subacromial bursa of the rats in the lentivirus‑RNAi group and lentivirus‑NC group, whereas 100 µl PBS was injected at the same site in the SAB group and the PBS blank control group. At 5 weeks following injection, the animals were sacrificed and venous blood was obtained. The effect of TNF‑α interference and the expression of inflammatory cytokines were determined by reverse transcription‑quantitative polymerase chain reaction, western blotting, hematoxylin and eosin staining, Van Gieson's staining and immunofluorescence. The expression of TNF‑α was decreased in the lentivirus‑TNF‑α‑RNAi group compared with that in the SAB group. Morphological observations revealed that the number of inflammatory cells were reduced and damage to tendon fibers was attenuated in this group, suggesting that the downregulation of the protein expression levels of TNF‑α‑associated nuclear

  8. Transgenic tobacco plants expressing siRNA targeted against the Mungbean yellow mosaic virus transcriptional activator protein gene efficiently block the viral DNA accumulation.

    PubMed

    Shanmugapriya, Gnanasekaran; Das, Sudhanshu Sekhar; Veluthambi, Karuppannan

    2015-06-01

    Mungbean yellow mosaic virus (MYMV) is a bipartite begomovirus that infects many pulse crops such as blackgram, mungbean, mothbean, Frenchbean, and soybean. We tested the efficacy of the transgenically expressed intron-spliced hairpin RNA gene of the transcriptional activator protein (hpTrAP) in reducing MYMV DNA accumulation. Tobacco plants transformed with the MYMV hpTrAP gene accumulated 21-22 nt siRNA. Leaf discs of the transgenic plants, agroinoculated with the partial dimers of MYMV, displayed pronounced reduction in MYMV DNA accumulation. Thus, silencing of the TrAP gene, a suppressor of gene silencing, emerged as an effective strategy to control MYMV.

  9. Junction formation and current transport mechanisms in hybrid n-Si/PEDOT:PSS solar cells

    NASA Astrophysics Data System (ADS)

    Jäckle, Sara; Mattiza, Matthias; Liebhaber, Martin; Brönstrup, Gerald; Rommel, Mathias; Lips, Klaus; Christiansen, Silke

    2015-08-01

    We investigated hybrid inorganic-organic solar cells combining monocrystalline n-type silicon (n-Si) and a highly conductive polymer poly(3,4-ethylenedioxythiophene)-poly(styrene sulfonate) (PEDOT:PSS). The build-in potential, photo- and dark saturation current at this hybrid interface are monitored for varying n-Si doping concentrations. We corroborate that a high build-in potential forms at the hybrid junction leading to strong inversion of the n-Si surface. By extracting work function and valence band edge of the polymer from ultraviolet photoelectron spectroscopy, a band diagram of the hybrid n-Si/PEDOT:PSS heterojunction is presented. The current-voltage characteristics were analyzed using Schottky and abrupt pn-junction models. The magnitude as well as the dependence of dark saturation current on n-Si doping concentration proves that the transport is governed by diffusion of minority charge carriers in the n-Si and not by thermionic emission of majorities over a Schottky barrier. This leads to a comprehensive explanation of the high observed open-circuit voltages of up to 634 mV connected to high conversion efficiency of almost 14%, even for simple planar device structures without antireflection coating or optimized contacts. The presented work clearly shows that PEDOT:PSS forms a hybrid heterojunction with n-Si behaving similar to a conventional pn-junction and not, like commonly assumed, a Schottky junction.

  10. Junction formation and current transport mechanisms in hybrid n-Si/PEDOT:PSS solar cells

    PubMed Central

    Jäckle, Sara; Mattiza, Matthias; Liebhaber, Martin; Brönstrup, Gerald; Rommel, Mathias; Lips, Klaus; Christiansen, Silke

    2015-01-01

    We investigated hybrid inorganic-organic solar cells combining monocrystalline n-type silicon (n-Si) and a highly conductive polymer poly(3,4-ethylenedioxythiophene)-poly(styrene sulfonate) (PEDOT:PSS). The build-in potential, photo- and dark saturation current at this hybrid interface are monitored for varying n-Si doping concentrations. We corroborate that a high build-in potential forms at the hybrid junction leading to strong inversion of the n-Si surface. By extracting work function and valence band edge of the polymer from ultraviolet photoelectron spectroscopy, a band diagram of the hybrid n-Si/PEDOT:PSS heterojunction is presented. The current-voltage characteristics were analyzed using Schottky and abrupt pn-junction models. The magnitude as well as the dependence of dark saturation current on n-Si doping concentration proves that the transport is governed by diffusion of minority charge carriers in the n-Si and not by thermionic emission of majorities over a Schottky barrier. This leads to a comprehensive explanation of the high observed open-circuit voltages of up to 634 mV connected to high conversion efficiency of almost 14%, even for simple planar device structures without antireflection coating or optimized contacts. The presented work clearly shows that PEDOT:PSS forms a hybrid heterojunction with n-Si behaving similar to a conventional pn-junction and not, like commonly assumed, a Schottky junction. PMID:26278010

  11. SI Notes.

    ERIC Educational Resources Information Center

    Nelson, Robert A.

    1983-01-01

    Discusses legislation related to SI (International Systems of Units) in the United States. Indicates that although SI metric units have been officially recognized by law in the United States, U.S. Customary Units have never received a statutory basis. (JN)

  12. Ordered CaSi2 Microwall Arrays on Si Substrates Induced by the Kirkendall Effect.

    PubMed

    Meng, Xiang; Ueki, Akiko; Tatsuoka, Hirokazu; Itahara, Hiroshi

    2017-03-02

    We have specified the synthetic conditions to obtain one-directionally ordered CaSi2 microwall arrays vertically grown on a Si substrate. Our basic concept is based on the utilization of the Kirkendall effect for reactive deposition epitaxy (RDE). We found for the first time that: 1) a much larger Ca vapor supply on the Si substrate than the conventional RDE, 2) the adoption of a two-step heating process, and 3) the selection of the crystal axis of the Si surface are the keys to control the microstructures of CaSi2 on the Si substrate. The CaSi phase was first formed on Si, then the CaSi2 phase was formed at the CaSi/Si interface. Based on the Kirkendall effect, the interdiffusion of Ca and Si was enhanced in the vertical direction rather than in the parallel direction to the Si surface. CaSi2 tends to grow along four equivalent Si{111} planes, however, the specific orientation of the Si surface resulted in CaSi2 microwalls grown along its Si(111‾ ) plane, the only plane directing nearly vertical to the surface among the Si{111} planes. These results suggest that the Kirkendall effect under asymmetric growth of target materials would be a rational strategy to obtain their ordered microstructures. © 2017 Wiley-VCH Verlag GmbH & Co. KGaA, Weinheim.

  13. Quantitative measurement of active dopant density distribution in phosphorus-implanted monocrystalline silicon solar cell using scanning nonlinear dielectric microscopy

    NASA Astrophysics Data System (ADS)

    Hirose, K.; Tanahashi, K.; Takato, H.; Cho, Y.

    2017-07-01

    The performance of silicon (Si) solar cells is dependent on the active dopant distribution in emitters. Estimation of the dopant distribution in a Si solar cell fabricated by ion implantation is difficult due to the pyramidal surface texture of the emitter. Here, we investigate the active dopant distribution in a P-implanted Si solar cell using scanning nonlinear dielectric microscopy (SNDM). SNDM and dC/dz-SNDM are complementarily applied to visualize the carrier distribution in the cross section of the Si solar cell. The carrier density in the emitter is calibrated using the SNDM data obtained from Si standard samples. The results show that the dopant distribution can be described as the sum of two Gaussian functions distributed over the vertical direction of the pyramidal faces. The three-dimensional (3D) dopant distribution is estimated from the superposition of the P distributions at each pyramidal face. The estimated 3D dopant distribution is in good agreement with the SNDM measurement results.

  14. Functionalized liposomes loaded with siRNAs targeting ion channels in effector memory T cells as a potential therapy for autoimmunity

    PubMed Central

    Hajdu, Péter; Chimote, Ameet A.; Thompson, Tyler; Koo, Youngmi; Yun, Yeoheung; Conforti, Laura

    2013-01-01

    Effector memory T cells (TM) play a key role in the pathology of certain autoimmune disorders. The activity of effector TM cells is under the control of Kv1.3 ion channels, which facilitate the Ca2+ influx necessary for T cell activation and function, i.e. cytokine release and proliferation. Consequently, the knock-down of Kv1.3 expression in effector TM’s may be utilized as a therapy for the treatment of autoimmune diseases. In this study we synthesized lipid unilamellar nanoparticles (NPs) that can selectively deliver Kv1.3 siRNAs into TM cells in vitro. NPs made from a mixture of phosphatidylcholine, pegylated/biotinylated phosphoethanolamine and cholesterol were functionalized with biotinylated-CD45RO (cell surface marker of TM’s) antibodies via fluorophore-conjugated streptavidin (CD45RO-NPs). Incubation of T cells with CD45RO-NPs resulted into the selective attachment and endocytosis of the NPs into TM’s. Furthermore, the siRNA against Kv1.3, encapsulated into the CD45RO-NPs, was released into the cytosol. Consequently, the expression of Kv1.3 channels decreased significantly in TM’s, which led to a remarkable decrease in Ca2+ influx. Our results can form the basis of an innovative therapeutic approach in autoimmunity. PMID:24075407

  15. Physical and Structural Characterization of a Monocrystalline Cu-13.7Al-4.2Ni Alloy Subjected to Thermal Cycling Treatments

    NASA Astrophysics Data System (ADS)

    Pereira, Elaine Cristina; Matlakhova, Lioudmila Aleksandrovna; Matlakhov, Anatoliy Nikolaevich; Shigue, Carlos Yujiro; Monteiro, Sérgio Neves

    2014-04-01

    A monocrystalline alloy with nominal 82wt pctCu-13.7wt pctAl-4.2wt pctNi composition and exhibiting reversible martensitic transformation (RMT) was subjected to multiple heating and cooling cycles within the RMT range of critical temperatures. Both untreated and cyclic treated alloy samples were characterized by X-ray diffraction, optical microscopy, differential scanning calorimetry, and Vickers microhardness. The results indicated that the alloy presents a complex RMT behavior disclosing a sequence of transformation steps: β 1 ↔ R and R ↔ β'1 + γ'1 as well as possible β 1 ↔ β'1 and β'1 ↔ γ'1 direct reactions. The thermal cycling treatment inhibits the development of γ'1 martensite without much change in both the physical and microstructure characteristics. This suggests a good resistance of the alloy to irreversible structural changes.

  16. Structural and photoluminescent properties of nanowires formed by the metal-assisted chemical etching of monocrystalline silicon with different doping level

    SciTech Connect

    Georgobiani, V. A. Gonchar, K. A.; Osminkina, L. A.; Timoshenko, V. Yu.

    2015-08-15

    Silicon-nanowire layers grown by the metal-assisted chemical etching of (100)-oriented p-type monocrystalline silicon substrates with a resistivity of 10 and 0.001 Ω · cm are studied by electron microscopy, Raman scattering, and photoluminescence measurements. It is established that nanowires grown on lightly doped substrates are structurally nonporous and formed as crystalline cores covered by nanocrystals 3–5 nm in dimensions. Nanowires grown on heavily doped substrates are structurally porous and contain both small nanocrystals and coarser crystallites with equilibrium charge carriers that influence interband radiative recombination. It is found that the photoluminescence intensity of nanowires in the spectral range 1.3–2.0 eV depends on the presence of molecular oxygen.

  17. Functionalized PEI Nanoparticles For Delivery Of IGF-1R-Targeted siRNA's to UPAR-Expressing Tumors In Vitro And In Vivo

    SciTech Connect

    Giblin, Michael F

    2012-12-14

    This proposal addressed the use of imaging technologies to develop therapeutic nanoparticle constructs which could reduce expression of molecules within the cancer cell important in tumor progression. The proposal described new labeling techniques that would result in therapeutic constructs which could be tracked both within targeted cells individually as well as within the individuals being treated. Representing a new generation of dual-labeled in vivo imaging agent, the constructs envisioned here would allow microPET imaging of targeted receptor expression as well as fluorescent imaging of silencing complexes targeting IGF-1R mRNA's. As such, this proposal was highly relevant to the Office of Biological and Environmental Research (BER) goals of facilitating improvements in radiotracer design in order to solve critical problems in biology and nuclear medicine.

  18. siRNA capsulated brain-targeted nanoparticles specifically knock down OATP2B1 in mice: a mechanism for acute morphine tolerance suppression

    PubMed Central

    Yang, Zi-Zhao; Li, Li; Wang, Lu; Xu, Ming-Cheng; An, Sai; Jiang, Chen; Gu, Jing-Kai; Wang, Zai-Jie Jim; Yu, Lu-Shan; Zeng, Su

    2016-01-01

    Regulating main brain-uptake transporter of morphine may restrict its tolerance generation, then modify its antinociception. In this study, more than 2 fold higher intracellular uptake concentrations for morphine and morphine-6-glucuronide (M6G) were observed in stable expression cells, HEK293-hOATP2B1 than HEK293-MOCK. Specifically, the Km value of morphine to OATP2B1 (57.58 ± 8.90 μM) is 1.4-time more than that of M6G (80.31 ± 21.75 μM); Cyclosporine A (CsA), an inhibitor of OATP2B1, can inhibit their intracellular accumulations with IC50 = 3.90 ± 0.50 μM for morphine and IC50 = 6.04 ± 0.86 μM for M6G, respectively. To further investigate the role of OATP2B1 in morphine brain transport and tolerance, the novel nanoparticles of DGL-PEG/dermorphin capsulated siRNA (OATP2B1) were applied to deliver siRNA into mouse brain. Along with OATP2B1 depressed, a main reduction was found for each of morphine or M6G in cerebrums or epencephalons of acute morphine tolerance mice. Furthermore, calcium/calmodulin-dependent protein kinase IIα (CaMKIIα) in mouse prefrontal cortex (mPFC) underwent dephosphorylation at Thr286. In conclusion, OATP2B1 downregulation in mouse brain can suppress tolerance via blocking morphine and M6G brain transport. These findings might help to improve the pharmacological effects of morphine. PMID:27629937

  19. Target-triggered signal turn-on detection of prostate specific antigen based on metal-enhanced fluorescence of Ag@SiO2@SiO2-RuBpy composite nanoparticles

    NASA Astrophysics Data System (ADS)

    Deng, Yun-Liang; Xu, Dang-Dang; Pang, Dai-Wen; Tang, Hong-Wu

    2017-02-01

    A three-layer core-shell nanostructure consisting of a silver core, a silica spacer, and a fluorescent dye RuBpy-doped outer silica layer was fabricated, and the optimal metal-enhanced fluorescence (MEF) distance was explored through adjusting the thickness of the silica spacer. The results show that the optimal distance is ˜10.4 nm with the maximum fluorescence enhancement factor 2.12. Then a new target-triggered MEF ‘turn-on’ strategy based on the optimized composite nanoparticles was successfully constructed for quantitative detection of prostate specific antigen (PSA), by using RuBpy as the energy donor and BHQ-2 as the acceptor. The hybridization of the complementary DNA of PSA-aptamer immobilized on the surface of the MEF nanoparticles with PSA-aptamer modified with BHQ-2, brought BHQ-2 in close proximity to RuBpy-doped silica shell and resulted in the decrease of fluorescence. In the presence of target PSA molecules, the BHQ-PSA aptamer is dissociated from the surface of the nanoparticles with the fluorescence switched on. Therefore, the assay of PSA was achieved by measuring the varying fluorescence intensity. The results show that PSA can be detected in the range of 1-100 ng ml-1 with a detection limit of 0.20 ng ml-1 (6.1 pM), which is 6.7-fold increase of that using hollow RuBpy-doped silica nanoparticles. Moreover, satisfactory results were obtained when PSA was detected in 1% serum.

  20. Menin missense mutants encoded by the MEN1 gene that are targeted to the proteasome: restoration of expression and activity by CHIP siRNA.

    PubMed

    Canaff, Lucie; Vanbellinghen, Jean-François; Kanazawa, Ippei; Kwak, Hayeon; Garfield, Natasha; Vautour, Line; Hendy, Geoffrey N

    2012-02-01

    In multiple endocrine neoplasia type 1 (MEN1) characterized by tumors of parathyroid, enteropancreas, and anterior pituitary, missense mutations in the MEN1 gene product, menin, occur in a subset of cases. The mutant proteins are degraded by the proteasome. However, whether their expression and activity can be restored is not known. Our objective was to functionally characterize a panel of 16 menin missense mutants, including W423R and S443Y identified in new MEN1 families, with respect to protein stability, targeting to the proteasome and restoration of expression by proteasome inhibitors and expression and function by small interfering RNA technology. Flag-tagged wild-type (WT) and missense menin mutant expression vectors were transiently transfected in human embryonic kidney (HEK293) and/or rat insulinoma (Rin-5F) cells. The majority of mutants were short-lived, whereas WT menin was stable. Proteasome inhibitors MG132 and PS-341 and inhibition of the chaperone, heat-shock protein 70 (Hsp70), or the ubiquitin ligase, COOH terminus of Hsp70-interacting protein (CHIP), by specific small interfering RNA, restored the levels of the mutants, whereas that of WT menin was largely unaffected. Inhibition of CHIP restored the ability of mutants to mediate normal functions of menin: TGF-β up-regulation of the promoters of its target genes, the cyclin-dependent kinase inhibitors p15 and p21 as well as TGF-β inhibition of cell numbers. When the levels of missense menin mutants that are targeted to the proteasome are normalized they may function similarly to WT menin. Potentially, targeting specific components of the proteasome chaperone pathway could be beneficial in treating a subset of MEN1 cases.

  1. Assessment of Nanobiotechnology-Targeted siRNA Designed to Inhibit NF-kappaB Classical And Alternative Signaling in Breast Tumor Macrophages

    DTIC Science & Technology

    2012-07-01

    including mannose, fucose , and galactose via ‘click’ chemistry. In this work, we show that the incorporation of contrast agents into the...following blocks: (1) an azide-displaying block for the attachment of alkyne-functionalized mannose, fucose , or galactose via ‘click’ chemistry, (2) a...immunohistochemistry). Biodistribution studies involving the fucose - and galactose-targeted nanoparticles are ongoing. Conclusions: The materials developed

  2. Tumor-targeted Nanoparticle Delivery of HuR siRNA Inhibits Lung Tumor Growth In Vitro and In Vivo By Disrupting the Oncogenic Activity of the RNA-binding Protein HuR.

    PubMed

    Muralidharan, Ranganayaki; Babu, Anish; Amreddy, Narsireddy; Srivastava, Akhil; Chen, Allshine; Zhao, Yan Daniel; Kompella, Uday B; Munshi, Anupama; Ramesh, Rajagopal

    2017-08-01

    Selective downregulation of the human antigen R (HuR) protein by siRNA may provide a powerful approach for treating lung cancer. To this end, we investigated the efficacy of transferrin receptor-targeted liposomal nanoparticle-based HuR siRNA (HuR-TfNP) therapy and compared with control siRNA (C)-TfNP therapy both, in vitro and in vivo using lung cancer models. In vitro studies showed HuR-TfNP, but not C-TfNP, efficiently downregulated HuR and HuR-regulated proteins in A549, and HCC827 lung cancer cells, resulting in reduced cell viability, inhibition of cell migration and invasion, and induction of G1 cell-cycle arrest culminating in apoptosis. However, HuR-TfNP activity in normal MRC-9 lung fibroblasts was negligible. In vivo biodistribution study demonstrated that fluorescently labeled HuR-siRNA or ICG dye-loaded TfNP localized in tumor tissues. Efficacy studies showed intratumoral or intravenous administration of HuR-TfNP significantly inhibited A549 (>55% inhibition) and HCC827 (>45% inhibition) subcutaneous tumor growth compared with C-TfNP. Furthermore, HuR-TfNP treatment reduced HuR, Ki67, and CD31 expression and increased caspase-9 and PARP cleavage and TUNEL-positive staining indicative of apoptotic cell death in tumor tissues compared with C-TfNP treatment. The antitumor activity of HuR-TfNP was also observed in an A549-luc lung metastatic model, as significantly fewer tumor nodules (9.5 ± 3.1; P < 0.001; 88% inhibition) were observed in HuR-TfNP-treated group compared with the C-TfNP-treated group (77.7 ± 20.1). Significant reduction in HuR, Ki67, and CD31 expression was also observed in the tumor tissues of HuR-TfNP-treatment compared with C-TfNP treatment. Our findings highlight HuR-TfNP as a promising nanotherapeutic system for lung cancer treatment. Mol Cancer Ther; 16(8); 1470-86. ©2017 AACR. ©2017 American Association for Cancer Research.

  3. Hollow carbon nanobubbles: monocrystalline MOF nanobubbles and their pyrolysis† †Electronic supplementary information (ESI) available. See DOI: 10.1039/c6sc04903f Click here for additional data file.

    PubMed Central

    Zhang, Wei; Jiang, Xiangfen; Zhao, Yanyi; Carné-Sánchez, Arnau; Malgras, Victor; Kim, Jeonghun; Kim, Jung Ho; Wang, Shaobin; Jiang, Ji-Sen

    2017-01-01

    While bulk-sized metal–organic frameworks (MOFs) face limits to their utilization in various research fields such as energy storage applications, nanoarchitectonics is believed to be a possible solution. It is highly challenging to realize MOF nanobubbles with monocrystalline frameworks. By a spatially controlled etching approach, here, we can achieve the synthesis of zeolitic imidazolate framework (ZIF-8) nanobubbles with a uniform size of less than 100 nm. Interestingly, the ZIF-8 nanobubbles possess a monocrystalline nanoshell with a thickness of around 10 nm. Under optimal pyrolytic conditions, the ZIF-8 nanobubbles can be converted into hollow carbon nanobubbles while keeping their original shapes. The structure of the nanobubble enhances the fast Na+/K+ ion intercalation performance. Such remarkable improvement cannot be realized by conventional MOFs or their derived carbons. PMID:28580098

  4. High-k gadolinium scandate on Si obtained by high pressure sputtering from metal targets and in-situ plasma oxidation

    NASA Astrophysics Data System (ADS)

    Pampillón, M. A.; San Andrés, E.; Feijoo, P. C.; Fierro, J. L. G.

    2017-03-01

    This article studies the physical and electrical behavior of Gd2‑x Sc x O3 layers grown by high pressure sputtering from metallic Gd and Sc targets. The aim is to obtain a high permittivity dielectric for microelectronic applications. The films were obtained by the deposition of a metallic nanolaminate of Gd and Sc alternating layers, which is afterwards in-situ oxidized by plasma. The oxide films obtained were close to stoichiometry, amorphous and with minimal interfacial regrowth. By fabricating metal–insulator–semiconductor capacitors we found that a moderate temperature annealing is needed to enhance permittivity, which reaches a high value of 32 while keeping moderate leakage. Finally, the feasibility of interface scavenging in this material with Ti gate electrodes is also demonstrated.

  5. Characterization of Si electrode-electrolyte interface under the illumination

    NASA Astrophysics Data System (ADS)

    Jiang, H. X.

    The flatband potential (Vfb) and the capacitance of n- and p-type Si used as an electrode in an electrolyte were examined experimentally under various intensities of illumination. The p-type material had an acceptor density of 8.0 x 10 to the 16th/cu cm, while the n-type Si acceptor density was 1.8 x 10 to the 17th/cu cm. The electrolyte was a 0.3 M NaCl solution and the interface areas with both types of monocrystalline wafer electrodes were about 0.5 sq cm. Reference measurements were made with a saturated calomel electrode (SCE) and a Pt wire cathode. The electrodes were biased and exposed to light from a 100 W tungsten lamp through a glass window. Measurements were made of the capacitance with respect to the recovery time, the variations of capacitance with light intensity, and the capacitance of the interface as a function of the light intensity. The data permitted definition of numerical models the total space charge per unit area of the electrode surface and for predicting the capacitance of the interface.

  6. TiO2 thin and thick films grown on Si/glass by sputtering of titanium targets in an RF inductively coupled plasma

    NASA Astrophysics Data System (ADS)

    Valencia-Alvarado, R.; de la Piedad-Beneitez, A.; López-Callejas, R.; Mercado-Cabrera, A.; Peña-Eguiluz, R.; Muñoz-Castro, A. E.; Rodríguez-Méndez, B. G.; de la Rosa-Vázquez, J. M.

    2015-03-01

    TiO2 thin and thick films were deposited on silicon/glass substrates using RF inductive plasma in continuous wave. The films thickness, as well as phases control, is achieved with a gradual increase in temperature substrates varying supplied RF power or working gas pressure besides deposition time as well. The deposition conditions were: argon 80%/oxygen 20% carefully calibrated mixture of 2 to 7×10-2 mbar as working gas pressure range. Deposition time 0.5 to 5 hours, 500 or 600 W RF power at 13.56 MHz frequency and 242-345 °C substrates temperature range. The titanium dioxide deposited on the substrates is grown by sputtering of a titanium target negatively polarized at 3-5 kV DC situated 14 mm in front of such substrates. The plasma reactor is a simple Pyrex-like glass cylindrical vessel of 50 cm long and 20 cm in diameter. Using the before describe plasma parameters we obtained films only anatase and both anatase/rutile phases with stoichiometric different. The films were characterized by X-ray photoelectron spectroscopy (XPS), stylus profilometer, X-ray diffraction (XRD), scanning electron microscopy (SEM) and Raman spectroscopy.

  7. Effects of vibration frequency on vibration-assisted nano-scratch process of mono-crystalline copper via molecular dynamics simulation

    SciTech Connect

    Zhu, Bo; Zhao, Hongwei E-mail: khl69@163.com; Zhao, Dan; Zhang, Peng; Yang, Yihan; Han, Lei; Kui, Hailin E-mail: khl69@163.com

    2016-03-15

    It has always been a critical issue to understand the material removal behavior of Vibration-Assisted Machining (VAM), especially on atomic level. To find out the effects of vibration frequency on material removal response, a three-dimensional molecular dynamics (MD) model has been established in this research to investigate the effects of scratched groove, crystal defects on the surface quality, comparing with the Von Mises shear strain and tangential force in simulations during nano-scratching process. Comparisons are made among the results of simulations from different vibration frequency with the same scratching feed, depth, amplitude and crystal orientation. Copper potential in this simulation is Embedded-Atom Method (EAM) potential. Interaction between copper and carbon atoms is Morse potential. Simulational results show that higher frequency can make groove smoother. Simulation with high frequency creates more dislocations to improve the machinability of copper specimen. The changing frequency does not have evident effects on Von Mises shear strain. Higher frequency can decrease the tangential force to reduce the consumption of cutting energy and tool wear. In conclusion, higher vibration frequency in VAM on mono-crystalline copper has positive effects on surface finish, machinablility and tool wear reduction.

  8. NMR evidence for an intimate relationship between antiferromagnetic spin fluctuations and extended s -wave superconductivity in monocrystalline SrFe2(As1-xPx) 2

    NASA Astrophysics Data System (ADS)

    Miyamoto, M.; Mukuda, H.; Kobayashi, T.; Yashima, M.; Kitaoka, Y.; Miyasaka, S.; Tajima, S.

    2015-09-01

    We report on a systematic 31P -NMR study on an iron- (Fe-) based superconductor SrFe2(As1-xPx) 2 (Sr122AsP) in which a superconducting (SC) transition temperature Tc at x =0.35 increases from Tc=26 K up to 33 K by annealing an as-grown monocrystalline sample. The present NMR study has unraveled that Tc reaches the highest value of 33 K at x =0.35 around a quantum critical point at which antiferromagnetic (AFM) order disappears. When noting that the SC transition disappears at x =0.6 where the AFM spin fluctuations (SFs) are no longer present, we remark that the onset and increase in Tc are apparently associated with the emergence and enhancement, respectively, of the AFM-SFs. In the SC state, the residual density of state (RDOS) at the Fermi energy EF in the SC state becomes much smaller for the annealed sample than for the as-grown one, suggesting that some inhomogeneity and/or imperfection for the latter increases the RDOS as expected for the unconventional SC state with a nodal gap. These findings in Sr122AsP are consistent with the unconventional s±-wave Cooper pairing state that is mediated by the AFM-SFs. We also discuss other key ingredients besides the AFM-SFs to increase Tc further.

  9. A molecular dynamics investigation into the mechanisms of subsurface damage and material removal of monocrystalline copper subjected to nanoscale high speed grinding

    NASA Astrophysics Data System (ADS)

    Li, Jia; Fang, Qihong; Liu, Youwen; Zhang, Liangchi

    2014-06-01

    This paper investigates the mechanisms of subsurface damage and material removal of monocrystalline copper when it is under a nanoscale high speed grinding of a diamond tip. The analysis was carried out with the aid of three-dimensional molecular dynamics simulations. The key factors that would influence the deformation of the material were carefully explored by analyzing the chip, dislocation movement, and workpiece deformation, which include grinding speed, depth of cut, grid tip radius, crystal orientation and machining angle of copper. An analytical model was also established to predict the emission of partial dislocations during the nanoscale high speed grinding. The investigation showed that a higher grinding velocity, a larger tip radius or a larger depth of cut would result in a larger chipping volume and a greater temperature rise in the copper workpiece. A lower grinding velocity would produce more intrinsic stacking faults. It was also found that the transition of deformation mechanisms depends on the competition between the dislocations and deformation twinning. There is a critical machining angle, at which a higher velocity, a smaller tip radius, or a smaller depth of cut will reduce the subsurface damage and improve the smoothness of a ground surface. The established analytical model showed that the Shockley dislocation emission is most likely to occur with the crystal orientations of (0 0 1)[1 0 0] at 45° angle.

  10. Effects of vibration frequency on vibration-assisted nano-scratch process of mono-crystalline copper via molecular dynamics simulation

    NASA Astrophysics Data System (ADS)

    Zhu, Bo; Zhao, Hongwei; Zhao, Dan; Zhang, Peng; Yang, Yihan; Han, Lei; Kui, Hailin

    2016-03-01

    It has always been a critical issue to understand the material removal behavior of Vibration-Assisted Machining (VAM), especially on atomic level. To find out the effects of vibration frequency on material removal response, a three-dimensional molecular dynamics (MD) model has been established in this research to investigate the effects of scratched groove, crystal defects on the surface quality, comparing with the Von Mises shear strain and tangential force in simulations during nano-scratching process. Comparisons are made among the results of simulations from different vibration frequency with the same scratching feed, depth, amplitude and crystal orientation. Copper potential in this simulation is Embedded-Atom Method (EAM) potential. Interaction between copper and carbon atoms is Morse potential. Simulational results show that higher frequency can make groove smoother. Simulation with high frequency creates more dislocations to improve the machinability of copper specimen. The changing frequency does not have evident effects on Von Mises shear strain. Higher frequency can decrease the tangential force to reduce the consumption of cutting energy and tool wear. In conclusion, higher vibration frequency in VAM on mono-crystalline copper has positive effects on surface finish, machinablility and tool wear reduction.

  11. SEMICONDUCTOR TECHNOLOGY Texturization of mono-crystalline silicon solar cells in TMAH without the addition of surfactant

    NASA Astrophysics Data System (ADS)

    Weiying, Ou; Yao, Zhang; Hailing, Li; Lei, Zhao; Chunlan, Zhou; Hongwei, Diao; Min, Liu; Weiming, Lu; Jun, Zhang; Wenjing, Wang

    2010-10-01

    Etching was performed on (100) silicon wafers using silicon-dissolved tetramethylammonium hydroxide (TMAH) solutions without the addition of surfactant. Experiments were carried out in different TMAH concentrations at different temperatures for different etching times. The surface phenomena, etching rates, surface morphology and surface reflectance were analyzed. Experimental results show that the resulting surface covered with uniform pyramids can be realized with a small change in etching rates during the etching process. The etching mechanism is explained based on the experimental results and the theoretical considerations. It is suggested that all the components in the TMAH solutions play important roles in the etching process. Moreover, TMA+ ions may increase the wettability of the textured surface. A good textured surface can be obtained in conditions where the absorption of OH-/H2O is in equilibrium with that of TMA+/SiO2 (OH)22-.

  12. Crystalline (Al1-xBx) PSi3 and (Al1-xBx) AsSi3 tetrahedral phases via reaction of Al(BH4)3 and M(SiH3)3 (M =P, As)

    NASA Astrophysics Data System (ADS)

    Sims, Patrick; White, Andrew; Aoki, Toshihiro; Menendez, Jose; Kouvetakis, John

    2015-03-01

    Crystalline (Al1-xBx) PSi3 alloys (x = 0.04-0.06) are grown lattice-matched on Si(100) by reactions of P(SiH3)3 and Al(BH4)3 using low-pressure CVD. The materials have been characterized by ellipsometry, XRD, XTEM, EELS and EDS, indicating the formation of single-phase monocrystalline layers with tetrahedral structures based on AlPSi3. The latter comprises interlinked AlPSi3 tetrahedra in which Al-P pairs are isolated within a Si matrix. Raman scattering of Al1-xBxPSi3 films support the presence of substitutional B in place of Al and provides evidence that B is bonded to P. The substitution of B atoms is desirable for promoting lattice matching, as required for Si-based solar cell designs. Analogous reactions of As(SiH3)3 with Al(BH4)3 produce (Al1-xBx) AsSi3 in which the B incorporation is limited to doping concentrations at 1020 cm-3. In both cases the Al(BH4)3 efficiently delivers Al to create crystalline group IV-III-V materials comprising light, earth abundant elements with possible application in photovoltaics and light element refractory solids. Supported by NSF-DMR 1309090.

  13. Differential cross sections measurement of 28Si(p,p/γ)28Si and 29Si(p,p/γ)29Si reactions for PIGE applications

    NASA Astrophysics Data System (ADS)

    Jokar, A.; Kakuee, O.; Lamehi-Rachti, M.

    2016-03-01

    Differential cross sections for gamma-ray emission from the 28Si(p,p/γ)28Si (Eγ = 1779 keV) and the 29Si(p,p/γ)29Si (Eγ = 1273 keV) nuclear reactions were measured in the energy range of 2.0-3.2 MeV and 2.0-3.0 MeV, respectively. The thin Si targets were prepared by evaporating natural SiO onto self-supporting Ag films. The gamma-rays and backscattered protons were detected simultaneously. An HPGe detector placed at an angle of 90° with respect to beam direction was employed to collect gamma-rays while an ion implanted Si detector placed at a scattering angle of 165° was used to detect backscattered protons. The great advantage of this work is that differential cross sections were obtained with a procedure irrespective of absolute value of the collected beam charge.

  14. Crystallization of amorphous Si nanoclusters in SiO(x) films using femtosecond laser pulse annealings.

    PubMed

    Korchagina, T T; Gutakovsky, A K; Fedina, L I; Neklyudova, M A; Volodin, V A

    2012-11-01

    The SiO(x) films of various stoichiometries deposited on Si substrates with the use of the co-sputtering from two separate Si and SiO2 targets were annealed by femtosecond laser pulses. Femtosecond laser treatments were applied for crystallization of amorphous silicon nanoclusters in the silicon-rich oxide films. The treatments were carried out with the use of Ti-Sapphire laser with wavelength 800 nm and pulse duration about 30 fs. Regimes of crystallization of amorphous Si nanoclusters in the initial films were found. Ablation thresholds for SiO(x) films of various stoichiometries were discovered. The effect of laser assisted formation of a-Si nanoclusters in the non-stoichiometric dielectric films with relatively low concentration of additional Si atoms was also observed. This approach is applicable for the creation of dielectric films with semiconductor nanoclusters on non-refractory substrates.

  15. Sputtered Ta-Si-N diffusion barriers in Cu metallizations for Si

    NASA Technical Reports Server (NTRS)

    Kolawa, E.; Pokela, P. J.; Reid, J. S.; Chen, J. S.; Nicolet, Marc A.; Ruiz, R. P.

    1991-01-01

    Electrical measurements on shallow Si n+-p junction diodes with a 30-nm TiSi2 contacting layer demonstrate that an 80-nm-thick amorphous Ta36Si14N50 film prepared by reactive RF sputtering of a Ta5Si3 target in an Ar/N2 plasma very effectively prevents the interaction between the Si substrate with the TiSi2 contacting layer and a 500-nm Cu overlayer. The Ta36Si14N50 diffusion barrier maintains the integrity of the I-V characteristics up to 900 C for 30-min annealing in vacuum. It is concluded that the amorphous Ta36Si14N50 alloy is not only a material with a very low reactivity for copper, titanium, and silicon, but must have a small diffusivity for copper as well.

  16. Generation of siRNA Nanosheets for Efficient RNA Interference

    NASA Astrophysics Data System (ADS)

    Kim, Hyejin; Lee, Jae Sung; Lee, Jong Bum

    2016-04-01

    After the discovery of small interference RNA (siRNA), nanostructured siRNA delivery systems have been introduced to achieve an efficient regulation of the target gene expression. Here we report a new siRNA-generating two dimensional nanostructure in a formation of nanosized sheet. Inspired by tunable mechanical and functional properties of the previously reported RNA membrane, siRNA nanosized sheets (siRNA-NS) with multiple Dicer cleavage sites were prepared. The siRNA-NS has two dimensional structure, providing a large surface area for Dicer to cleave the siRNA-NS for the generation of functional siRNAs. Furthermore, downregulation of the cellular target gene expression was achieved by delivery of siRNA-NS without chemical modification of RNA strands or conjugation to other substances.

  17. Defect recognition by means of light and electron probe techniques for the characterization of mc-Si wafers and solar cells

    NASA Astrophysics Data System (ADS)

    Moralejo, B.; Tejero, A.; Hortelano, V.; Martínez, O.; González, M. A.; Jiménez, J.

    2016-11-01

    Multicristalline Silicon (mc-Si) is the preferred material for current terrestrial photovolt