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Sample records for double knudsen cell

  1. Double-Knudsen-Cell Apparatus Measures Alloy-Component Activities

    NASA Technical Reports Server (NTRS)

    Jacobson, Nathan S.; Moore, Warren A.

    1995-01-01

    Double-Knudsen-cell apparatus provides molecular beam from selected one of two Knudsen cells. Both cells maintained at same temperature. Molecular beam directed into mass spectrometer for measurement of vapor pressure of selected material component. Designed to minimize undesired thermal gradients, provides appropriate translation to place selected cell in position for sampling, and minimizes mixing of molecular beams from cells.

  2. Thermodynamic Measurements of Alloys and Compounds by Double Knudsen Cell Mass Spectrometry and Their Application to Materials Processing

    PubMed Central

    Sasaki, Hideaki; Kobashi, Yoshifumi; Nagai, Takashi; Maeda, Masafumi

    2014-01-01

    For the development and optimization of materials processing a collection of thermodynamic information concerning substances that participate in the reactions is important. One fundamental way to obtain such information is to measure the vapor pressure of gas species under conditions where they are in equilibrium with the condensed phases. Over the past 60 years Knudsen cell mass spectrometry has been used to identify and quantitatively determine gas species at high temperatures. This article describes thermodynamic foundation and examples of measurements in order to demonstrate the use of mass spectrometry focusing on the field of process metallurgy and recycling processes. PMID:26819884

  3. Twin Knudsen Cell Configuration for Activity Measurements by Mass Spectrometry

    NASA Technical Reports Server (NTRS)

    Jacobson, N. S.

    1996-01-01

    A twin Knudsen cell apparatus for alloy activity measurements by mass spectrometry is described. Two Knudsen cells - one containing an alloy and one containing a pure component - are mounted on a single flange and translated into the sampling region via a motorized x-y table. Mixing of the molecular beams from the cells is minimized by a novel system of shutters. Activity measurements were taken on two well-characterized alloys to verify the operation of the system. Silver activity measurements are reported for Ag-Cu alloys and aluminum activity measurements are reported for Fe-Al alloys. The temperature dependence of activity for a 0.474 mol fraction Al-Fe alloy gives a partial molar heat of aluminum. Measurements taken with the twin cell show good agreement with literature values for these alloys.

  4. Knudsen Cell Studies of Ti-Al Thermodynamics

    NASA Technical Reports Server (NTRS)

    Jacobson, Nathan S.; Copland, Evan H.; Mehrotra, Gopal M.; Auping, Judith; Gray, Hugh R. (Technical Monitor)

    2002-01-01

    In this paper we describe the Knudsen cell technique for measurement of thermodynamic activities in alloys. Numerous experimental details must be adhered to in order to obtain useful experimental data. These include introduction of an in-situ standard, precise temperature measurement, elimination of thermal gradients, and precise cell positioning. Our first design is discussed and some sample data on Ti-Al alloys is presented. The second modification and associated improvements are also discussed.

  5. Multiple Knudsen Cell Configuration Improved for Alloy Activity Studies

    NASA Technical Reports Server (NTRS)

    Copland, Evan H.; Jacobson, Nathan S.

    2003-01-01

    Knudsen effusion mass spectrometry (KEMS) allows the simultaneous determination of the identity and pressure of vapor species in equilibrium with a condensed phase as a function of temperature. This information can be used to determine the thermodynamic properties of materials. The partial pressure of species j in the cell is related to the measured intensity of the ion k formed from j, I(sup +) less than SABjk, and the absolute temperature T, where S(sub jk) is the sensitivity factor.

  6. Thermodynamic Activity Measurements with Knudsen Cell Mass Spectrometry

    NASA Technical Reports Server (NTRS)

    Copland, Evan H.; Jacobson, Nathan S.

    2001-01-01

    Coupling the Knudsen effusion method with mass spectrometry has proven to be one of the most useful experimental techniques for studying the equilibrium between condensed phases and complex vapors. The Knudsen effusion method involves placing a condensed sample in a Knudsen cell, a small "enclosure", that is uniformly heated and held until equilibrium is attained between the condensed and vapor phases. The vapor is continuously sampled by effusion through a small orifice in the cell. A molecular beam is formed from the effusing vapor and directed into a mass spectrometer for identification and pressure measurement of the species in the vapor phase. Knudsen cell mass spectrometry (KCMS) has been used for nearly fifty years now and continues to be a leading technique for obtaining thermodynamic data. Indeed, much of the well-established vapor specie data in the JANAF tables has been obtained from this technique. This is due to the extreme versatility of the technique. All classes of materials can be studied and all constituents of the vapor phase can be measured over a wide range of pressures (approximately 10(exp -4) to 10(exp -11) bar) and temperatures (500-2800 K). The ability to selectively measure different vapor species makes KCMS a very powerful tool for the measurement of component activities in metallic and ceramic solutions. Today several groups are applying KCMS to measure thermodynamic functions in multicomponent metallic and ceramic systems. Thermodynamic functions, especially component activities, are extremely important in the development of CALPHAD (Calculation of Phase Diagrams) type thermodynamic descriptions. These descriptions, in turn, are useful for modeling materials processing and predicting reactions such as oxide formation and fiber/matrix interactions. The leading experimental methods for measuring activities are the Galvanic cell or electro-motive force (EMF) technique and the KCMS technique. Each has specific advantages, depending on

  7. Improvements and new capabilities for the multiple Knudsen cell device used in high-temperature mass spectrometry.

    PubMed

    Heyrman, M; Chatillon, C; Collas, H; Chemin, J-L

    2004-01-01

    The thermodynamic properties of condensed phases, i.e. the activities of components, can be determined from partial pressures measured by the Knudsen cell mass spectrometric method. Improvements in accuracy and yield of this method are obtained with the use of twin cells, an idea proposed in the 1960s. The multiple cell method was perfected in 1977 in our laboratory. Changes to molecular beam sampling and furnace assemblies were required to make the multiple Knudsen cell technique work properly. This paper summarizes these prerequisites, and presents a new device and the associated method of measurements, as well as the necessary tests performed with a silver sample in each cell. PMID:14745765

  8. Thermodynamic Studies of High Temperature Materials Via Knudsen Cell Mass Spectrometry

    NASA Technical Reports Server (NTRS)

    Jacobson, Nathan S.; Brady, Michael P.

    1997-01-01

    The Knudsen Cell technique is a classic technique from high temperature chemistry for studying condensed phase/vapor equilibria. It is based on a small enclosure, usually about 1 cm in diameter by 1 cm high, with an orifice of well-defined geometry. This forms a molecular beam which is analyzed with mass spectrometry. There are many applications to both fundamental and applied problems with high temperature materials. Specific measurements include vapor pressures and vapor compositions above solids, activities of alloy components, and fundamental gas/solid reactions. The basic system is shown. Our system can accommodate a wide range of samples, temperatures, and attachments, such as gas inlets. It is one of only about ten such systems world-wide.

  9. Knudsen Cell Uptake Experiments of Oxides of Nitrogen on Fractional Layers of NaCl.

    NASA Astrophysics Data System (ADS)

    Hoffman, R. C.; Finlayson-Pitts, B. J.

    2002-12-01

    Sea salt aerosols rich in NaCl play important roles in heterogeneous chemistry occurring in the troposphere. Understanding the kinetics and mechanisms of these processes is essential to understanding the extent of the tropospheric halogen budget. Two such reactions involve HNO3 and N2O5 reacting with NaCl to generate photochemically inert HCl and photochemically active ClNO2, respectively. The uptake and reaction of HNO3 and N2O5 on NaCl and on synthetic sea salt (SSS) was studied using a Knudsen cell coupled to an electron impact quadrupole mass spectrometer. Experiments were conducted using fractional salt particle layers with average particle sizes of 164, 291, 312, and 428 μm. Fractional particle layers were used in order to minimize the uncertainty of the available reactive surface area due to diffusion of the reactant gas (either HNO3 or N2O5) into multiple salt layers. Reaction probabilities, found to differ by a factor of 2 between HNO3 and N2O5 for fractional salt layer experiments, will be presented and the atmospheric implications will be discussed.

  10. Idiosyncrasies of Physical Vapor Deposition Processes from Various Knudsen Cells for Quinacridone Thin Film Growth on Silicon Dioxide

    PubMed Central

    2015-01-01

    Thin films of quinacridone deposited by physical vapor deposition on silicon dioxide were investigated by thermal desorption spectroscopy (TDS), mass spectrometry (MS), atomic force microscopy (AFM), specular and grazing incidence X-ray diffraction (XRD, GIXD), and Raman spectroscopy. Using a stainless steel Knudsen cell did not allow the preparation of a pure quinacridone film. TDS and MS unambiguously showed that in addition to quinacridone, desorbing at about 500 K (γ-peak), significant amounts of indigo desorbed at about 420 K (β-peak). The existence of these two species on the surface was verified by XRD, GIXD, and Raman spectroscopy. The latter spectroscopies revealed that additional species are contained in the films, not detected by TDS. In the film mainly composed of indigo a species was identified which we tentatively attribute to carbazole. The film consisting of mainly quinacridone contained in addition p-sexiphenyl. The reason for the various decomposition species effusing from the metal Knudsen cell is the comparably high sublimation temperature of the hydrogen bonded quinacridone. With special experimental methods and by using glass Knudsen-type cells we were able to prepare films which exclusively consist of molecules either corresponding to the β-peak or the γ-peak. These findings are of relevance for choosing the proper deposition techniques in the preparation of quinacridone films in the context of organic electronic devices. PMID:26401189

  11. Knudsen cell and smog chamber study of the heterogeneous uptake of sulfur dioxide on Chinese mineral dust.

    PubMed

    Zhou, Li; Wang, Weigang; Gai, Yanbo; Ge, Maofa

    2014-12-01

    The heterogeneous uptake processes of sulfur dioxide on two types of Chinese mineral dust (Inner Mongolia desert dust and Xinjiang sierozem) were investigated using both Knudsen cell and smog chamber system. The temperature dependence of the uptake coefficients was studied over a range from 253 to 313 K using the Knudsen cell reactor, the initial uptake coefficients decreased with the increasing of temperature for these two mineral dust samples, whereas the steady state uptake coefficients of the Xinjiang sierozem increased with the temperature increasing, and these temperature dependence functions were obtained for the first time. In the smog chamber experiments at room temperature, the steady state uptake coefficients of SO2 decreased evidently with the increasing of sulfur dioxide initial concentration from 1.72 × 10¹² to 6.15 × 10¹² mol/cm³. Humid air had effect on the steady state uptake coefficients of SO₂onto Inner Mongolia desert dust. Consequences about the understanding of the uptake processes onto mineral dust samples and the environmental implication were also discussed. PMID:25499490

  12. Characterization of Evaporating Species from B2O3, B6O, and Their Mixtures by Knudsen Cell Mass Spectrometry

    NASA Astrophysics Data System (ADS)

    Sasaki, Hideaki; Kobashi, Yoshifumi; Maeda, Masafumi

    2016-02-01

    Species evaporating from B2O3(l), B6O(s) and their mixtures were observed by a multiple Knudsen cell mass spectrometer between 1373 K and 1573 K (1100 °C and 1300 °C). Ions with mass-to-charge ratios of 70, 54, and 27 from the samples were observable, indicating the formations of B2O3(g), B2O2(g), and BO(g). The vapor pressures of the gas species were estimated by referring to thermodynamic information previously reported on B6O(s). Evaporation of B2O2(g) from a mixture [ p_{B}_{2} O_{2} }} = 6 Pa at 1473 K (1200 °C)] was observed, and it was consistent with a preceding study by a different method.

  13. Knudsen heat capacity

    SciTech Connect

    Babac, Gulru; Reese, Jason M.

    2014-05-15

    We present a “Knudsen heat capacity” as a more appropriate and useful fluid property in micro/nanoscale gas systems than the constant pressure heat capacity. At these scales, different fluid processes come to the fore that are not normally observed at the macroscale. For thermodynamic analyses that include these Knudsen processes, using the Knudsen heat capacity can be more effective and physical. We calculate this heat capacity theoretically for non-ideal monatomic and diatomic gases, in particular, helium, nitrogen, and hydrogen. The quantum modification for para and ortho hydrogen is also considered. We numerically model the Knudsen heat capacity using molecular dynamics simulations for the considered gases, and compare these results with the theoretical ones.

  14. Synergistic use of Knudsen effusion quadrupole mass spectrometry, solid-state galvanic cell and differential scanning calorimetry for thermodynamic studies on lithium aluminates

    NASA Astrophysics Data System (ADS)

    Rakshit, S. K.; Naik, Y. P.; Parida, S. C.; Dash, Smruti; Singh, Ziley; Sen, B. K.; Venugopal, V.

    2008-06-01

    Three ternary oxides LiAl 5O 8(s), LiAlO 2(s) and Li 5AlO 4(s) in the system Li-Al-O were prepared by solid-state reaction route and characterized by X-ray powder diffraction method. Equilibrium partial pressure of CO 2(g) over the three-phase mixtures {LiAl 5O 8(s)+Li 2CO 3(s)+5Al 2O 3(s)}, {LiAl 5O 8(s)+5LiAlO 2(s)+2Li 2CO 3(s)} and {LiAlO 2(s)+Li 5AlO 4(s)+2Li 2CO 3(s)} were measured using Knudsen effusion quadrupole mass spectrometry (KEQMS). Solid-state galvanic cell technique based on calcium fluoride electrolyte was used to determine the standard molar Gibbs energies of formations of these aluminates. The standard molar Gibbs energies of formation of these three aluminates calculated from KEQMS and galvanic cell measurements were in good agreement. Heat capacities of individual ternary oxides were measured from 127 to 868 K using differential scanning calorimetry. Thermodynamic tables representing the values of Δ fH0(298.15 K), S0(298.15 K) S0( T), Cp0( T), H0( T), { H0( T)- H0(298.15 K)}, G0( T), Δ fH0( T), Δ fG 0( T) and free energy function (fef) were constructed using second law analysis and FACTSAGE thermo-chemical database software.

  15. Synergistic use of Knudsen effusion quadrupole mass spectrometry, solid-state galvanic cell and differential scanning calorimetry for thermodynamic studies on lithium aluminates

    SciTech Connect

    Rakshit, S.K. Naik, Y.P.; Parida, S.C.; Dash, Smruti; Singh, Ziley; Sen, B.K.; Venugopal, V.

    2008-06-15

    Three ternary oxides LiAl{sub 5}O{sub 8}(s), LiAlO{sub 2}(s) and Li{sub 5}AlO{sub 4}(s) in the system Li-Al-O were prepared by solid-state reaction route and characterized by X-ray powder diffraction method. Equilibrium partial pressure of CO{sub 2}(g) over the three-phase mixtures {l_brace}LiAl{sub 5}O{sub 8}(s)+Li{sub 2}CO{sub 3}(s)+5Al{sub 2}O{sub 3}(s){r_brace}, {l_brace}LiAl{sub 5}O{sub 8}(s)+5LiAlO{sub 2}(s)+2Li{sub 2}CO{sub 3}(s){r_brace} and {l_brace}LiAlO{sub 2}(s)+Li{sub 5}AlO{sub 4}(s)+2Li{sub 2}CO{sub 3}(s){r_brace} were measured using Knudsen effusion quadrupole mass spectrometry (KEQMS). Solid-state galvanic cell technique based on calcium fluoride electrolyte was used to determine the standard molar Gibbs energies of formations of these aluminates. The standard molar Gibbs energies of formation of these three aluminates calculated from KEQMS and galvanic cell measurements were in good agreement. Heat capacities of individual ternary oxides were measured from 127 to 868 K using differential scanning calorimetry. Thermodynamic tables representing the values of {delta}{sub f}H{sup 0}(298.15 K), S{sup 0}(298.15 K) S{sup 0}(T), C{sub p}{sup 0}(T), H{sup 0}(T), {l_brace}H{sup 0}(T)-H{sup 0}(298.15 K){r_brace}, G{sup 0}(T), {delta}{sub f}H{sup 0}(T), {delta}{sub f}G{sup 0}(T) and free energy function (fef) were constructed using second law analysis and FACTSAGE thermo-chemical database software. - Graphical abstract: Comparison of {delta}{sub f}G{sub m}{sup 0} of ternary oxides determined from KEQMS and solid-state galvanic cell techniques. (O) KEQMS, (9632;) solid-state galvanic cell and solid line: combined fit of both the experimental data.

  16. Knudsen Gas Provides Nanobubble Stability

    NASA Astrophysics Data System (ADS)

    Seddon, James R. T.; Zandvliet, Harold J. W.; Lohse, Detlef

    2011-09-01

    We provide a model for the remarkable stability of surface nanobubbles to bulk dissolution. The key to the solution is that the gas in a nanobubble is of Knudsen type. This leads to the generation of a bulk liquid flow which effectively forces the diffusive gas to remain local. Our model predicts the presence of a vertical water jet immediately above a nanobubble, with an estimated speed of ˜3.3m/s, in good agreement with our experimental atomic force microscopy measurement of ˜2.7m/s. In addition, our model also predicts an upper bound for the size of nanobubbles, which is consistent with the available experimental data.

  17. Molecular simulation of small Knudsen number flows

    NASA Astrophysics Data System (ADS)

    Fei, Fei; Fan, Jing

    2012-11-01

    The direct simulation Monte Carlo (DSMC) method is a powerful particle-based method for modeling gas flows. It works well for relatively large Knudsen (Kn) numbers, typically larger than 0.01, but quickly becomes computationally intensive as Kn decreases due to its time step and cell size limitations. An alternative approach was proposed to relax or remove these limitations, based on replacing pairwise collisions with a stochastic model corresponding to the Fokker-Planck equation [J. Comput. Phys., 229, 1077 (2010); J. Fluid Mech., 680, 574 (2011)]. Similar to the DSMC method, the downside of that approach suffers from computationally statistical noise. To solve the problem, a diffusion-based information preservation (D-IP) method has been developed. The main idea is to track the motion of a simulated molecule from the diffusive standpoint, and obtain the flow velocity and temperature through sampling and averaging the IP quantities. To validate the idea and the corresponding model, several benchmark problems with Kn ˜ 10-3-10-4 have been investigated. It is shown that the IP calculations are not only accurate, but also efficient because they make possible using a time step and cell size over an order of magnitude larger than the mean collision time and mean free path, respectively.

  18. Fusion utility in the Knudsen layer

    SciTech Connect

    Davidovits, Seth; Fisch, Nathaniel J.

    2014-09-15

    In inertial confinement fusion, the loss of fast ions from the edge of the fusing hot-spot region reduces the reactivity below its Maxwellian value. The loss of fast ions may be pronounced because of the long mean free paths of fast ions, compared with those of thermal ions. We introduce a fusion utility function to demonstrate essential features of this Knudsen layer effect, in both magnetized and unmagnetized cases. The fusion utility concept is also used to evaluate the restoring reactivity in the Knudsen layer by manipulating fast ions in phase space using waves.

  19. Fusion Utility in the Knudsen Layer

    SciTech Connect

    Davidovits, Seth; Fisch, Nathaniel J.

    2014-08-01

    In inertial confi nement fusion, the loss of fast ions from the edge of the fusing hot-spot region reduces the reactivity below its Maxwellian value. The loss of fast ions may be pronounced because of the long mean free paths of fast ions, compared to those of thermal ions. We introduce a fusion utility function to demonstrate essential features of this Knudsen layer e ffect, in both magnetized and unmagnetized cases. The fusion utility concept is also used to evaluate restoring the reactivity in the Knudsen layer by manipulating fast ions in phase space using waves.

  20. Occurrence of Knudsen minima in diverging microchannels

    SciTech Connect

    Hemadri, Vadiraj; Bhandarkar, Upendra; Agrawal, Amit

    2014-12-09

    Rarefied gas flow is gaining increasing importance with the emergence of Micro Electro Mechanical Systems (MEMS). Knudsen minima is one of the characteristic feature of such rarefied flows and has been observed in uniform cross section channels such as plane channel, cylindrical tube and annulus. However, data pertaining to gaseous flow in varying cross section channel is relatively sparse. Channels of varying cross section are frequently encountered in MEMS devices and are fundamental to the design of micro-scale nozzles and micro-valves. In this context, rarefied gas flow through a diverging microchannel (divergence angle – 12 degree) is studied experimentally with three different gases (argon, nitrogen and oxygen). The experiments are performed over a wide range with the mean Knudsen number varying from slip to the transitional regime (0.07 to 1.2). It is found that the effect of molecular weight of the gas on the non-dimensional mass flow rate is negligible. The Knudsen minima is experimentally observed for the first time in microchannel of non-uniform cross section.

  1. Knudsen torque on heated micro beams

    SciTech Connect

    Li, Qi; Liang, Tengfei; Ye, Wenjing

    2014-12-09

    Thermally induced mechanical loading has been shown to have significant effects on micro/nano objects immersed in a gas with a non-uniform temperature field. While the majority of existing studies and related applications focus on forces, we investigate the torque, and thus the rotational motion, produced by such a mechanism. Using the asymptotic analysis in the near continuum regime, the Knudsen torque acting on an asymmetrically located uniformly heated microbeam in a cold enclosure is investigated. The existence of a non-zero net torque is demonstrated. In addition, it has been found that by manipulating the system configuration, the rotational direction of the torque can be changed. Two types of rotational motion of the microbeam have been identified: the pendulum motion of a rectangular beam, and the unidirectional rotation of a cylindrical beam. A rotational frequency of 4 rpm can be achieved for the cylindrical beam with a diameter of 3μm at Kn = 0.005. Illustrated by the simulations using the direct simulation of Monte Carlo, the Knudsen torque can be much increased in the transition regime, demonstrating the potential of Knudsen torque serving as a rotation engine for micro/nano objects.

  2. Double-sided solar cell package

    NASA Technical Reports Server (NTRS)

    Shelpuk, B. (Inventor)

    1979-01-01

    In a solar cell array of terrestrial use, an improved double-sided solar cell package, consisting of a photovoltaic cell having a metallized P-contact strip and an N-contact grid, provided on opposite faces of the cell, a transparent tubular body forming an enclosure for the cell. A pedestal supporting the cell from within the enclosure comprising an electrical conductor connected with the P-contact strip provided for each face of the cell, and a reflector having an elongated reflective surface disposed in substantially opposed relation with one face of the cell for redirecting light were also included.

  3. Double interconnection fuel cell array

    DOEpatents

    Draper, Robert; Zymboly, Gregory E.

    1993-01-01

    A fuel cell array (10) is made, containing number of tubular, elongated fuel cells (12) which are placed next to each other in rows (A, B, C, D), where each cell contains inner electrodes (14) and outer electrodes (18 and 18'), with solid electrolyte (16 and 16') between the electrodes, where the electrolyte and outer electrode are discontinuous, having two portions, and providing at least two opposed discontinuities which contain at least two oppositely opposed interconnections (20 and 20') contacting the inner electrode (14), each cell (12) having only three metallic felt electrical connectors (22) which contact surrounding cells, where each row is electrically connected to the other.

  4. Double Stem Cell Transplant May Help Fight a Childhood Cancer

    MedlinePlus

    ... nih.gov/medlineplus/news/fullstory_159243.html Double Stem Cell Transplant May Help Fight a Childhood Cancer ... better chance of survival if they receive two stem cell transplants, a new study reports. The double ...

  5. Deviation from the Knudsen law on quantum gases

    SciTech Connect

    Babac, Gulru

    2014-12-09

    Gas flow in micro/nano scale systems has been generally studied for the Maxwell gases. In the limits of very low temperature and very confined domains, the Maxwellian approximation can break down and the quantum character of the gases becomes important. In these cases, Knudsen law, which is one of the important equations to analyze rarefied gas flows is invalid and should be reanalyzed for quantum gases. In this work, the availability of quantum gas conditions in the high Knudsen number cases is discussed and Knudsen law is analyzed for quantum gases.

  6. A diffusive information preservation method for small Knudsen number flows

    NASA Astrophysics Data System (ADS)

    Fei, Fei; Fan, Jing

    2013-06-01

    The direct simulation Monte Carlo (DSMC) method is a powerful particle-based method for modeling gas flows. It works well for relatively large Knudsen (Kn) numbers, typically larger than 0.01, but quickly becomes computationally intensive as Kn decreases due to its time step and cell size limitations. An alternative approach was proposed to relax or remove these limitations, based on replacing pairwise collisions with a stochastic model corresponding to the Fokker-Planck equation [J. Comput. Phys., 229, 1077 (2010); J. Fluid Mech., 680, 574 (2011)]. Similar to the DSMC method, the downside of that approach suffers from computationally statistical noise. To solve the problem, a diffusion-based information preservation (D-IP) method has been developed. The main idea is to track the motion of a simulated molecule from the diffusive standpoint, and obtain the flow velocity and temperature through sampling and averaging the IP quantities. To validate the idea and the corresponding model, several benchmark problems with Kn ˜ 10-3-10-4 have been investigated. It is shown that the IP calculations are not only accurate, but also efficient because they make possible using a time step and cell size over an order of magnitude larger than the mean collision time and mean free path, respectively.

  7. A diffusive information preservation method for small Knudsen number flows

    SciTech Connect

    Fei, Fei; Fan, Jing

    2013-06-15

    The direct simulation Monte Carlo (DSMC) method is a powerful particle-based method for modeling gas flows. It works well for relatively large Knudsen (Kn) numbers, typically larger than 0.01, but quickly becomes computationally intensive as Kn decreases due to its time step and cell size limitations. An alternative approach was proposed to relax or remove these limitations, based on replacing pairwise collisions with a stochastic model corresponding to the Fokker–Planck equation [J. Comput. Phys., 229, 1077 (2010); J. Fluid Mech., 680, 574 (2011)]. Similar to the DSMC method, the downside of that approach suffers from computationally statistical noise. To solve the problem, a diffusion-based information preservation (D-IP) method has been developed. The main idea is to track the motion of a simulated molecule from the diffusive standpoint, and obtain the flow velocity and temperature through sampling and averaging the IP quantities. To validate the idea and the corresponding model, several benchmark problems with Kn ∼ 10{sup −3}–10{sup −4} have been investigated. It is shown that the IP calculations are not only accurate, but also efficient because they make possible using a time step and cell size over an order of magnitude larger than the mean collision time and mean free path, respectively.

  8. Fundamental computation of molecular flux in Knudsen-regime nanopores

    NASA Astrophysics Data System (ADS)

    Colson, Fenner

    Molecular transport in systems of high Knudsen number is predominantly calculated using either Knudsen's equation or by Einstein's relation coupled with computer simulations. However, Knudsen's analysis assumes no adsorptive interaction between a diffusing molecule and the molecules of the pore wall and the majority of computer simulations apply non-physical methods to circumvent the computational expense of simulating enormous numbers of molecules. This research presents a fundamentally-derived formulation that preserves species-dependent wall effects and avoids introducing non-physical elements in the evaluation of molecular flux through Knudsen-regime nanopores. The analysis is based on the Knudsen assumption of negligible inter-molecular interactions of molecules diffusing through a constant area nanopore. Subsequent assumptions regarding steady-state and the functional shape of distribution functions simplify the flux expression into an extremely accessible form where straightforward yet realistic molecular dynamics simulations can provide the necessary parameters to evaluate flux and diffusion coefficients using the rigorously derived formula. Validation is completed analytically by showing that the general formulation reduces to existing theory under the appropriate simplifications, including the self-similarity solution of infinite diffusion, Fick's First Law, and Einstein's mean squared displacement relation.

  9. Amplification and reversal of Knudsen force by thermoelectric heating

    SciTech Connect

    O'Neill, William J.; Wada, Mizuki; Strongrich, Andrew D.; Cofer, Anthony; Alexeenko, Alina A.

    2014-12-09

    We show that the Knudsen thermal force generated by a thermally-induced flow over a heated beam near a colder wall could be amplified significantly by thermoelectric heating. Bidirectional actuation is achieved by switching the polarity of the thermoelectric device bias voltage. The measurements of the resulting thermal forces at different rarefaction regimes, realized by changing geometry and gas pressure, are done using torsional microbalance. The repulsive or attractive forces between a thermoelectrically heated or cooled plate and a substrate are shown to be up to an order of magnitude larger than for previously studied configurations and heating methods due to favorable coupling of two thermal gradients. The amplification and reversal of the Knudsen force is confirmed by numerical solution of the Boltzmann-ESBGK kinetic model equation. Because of the favorable scaling with decreasing system size, the Knudsen force with thermoelectric heating offers a novel actuation and sensing mechanism for nano/microsystems.

  10. Amplification and reversal of Knudsen force by thermoelectric heating

    NASA Astrophysics Data System (ADS)

    O'Neill, William J.; Wada, Mizuki; Strongrich, Andrew D.; Cofer, Anthony; Alexeenko, Alina A.

    2014-12-01

    We show that the Knudsen thermal force generated by a thermally-induced flow over a heated beam near a colder wall could be amplified significantly by thermoelectric heating. Bidirectional actuation is achieved by switching the polarity of the thermoelectric device bias voltage. The measurements of the resulting thermal forces at different rarefaction regimes, realized by changing geometry and gas pressure, are done using torsional microbalance. The repulsive or attractive forces between a thermoelectrically heated or cooled plate and a substrate are shown to be up to an order of magnitude larger than for previously studied configurations and heating methods due to favorable coupling of two thermal gradients. The amplification and reversal of the Knudsen force is confirmed by numerical solution of the Boltzmann-ESBGK kinetic model equation. Because of the favorable scaling with decreasing system size, the Knudsen force with thermoelectric heating offers a novel actuation and sensing mechanism for nano/microsystems.

  11. Slip effects associated with Knudsen transport phenomena in porous media

    NASA Technical Reports Server (NTRS)

    Frederking, T. H. K.; Hepler, W. A.; Khandhar, P. K.

    1988-01-01

    Porous media used in phase separators and thermomechanical pumps have been the subject of characterization efforts based on the Darcy permeability of laminar continuum flow. The latter is not always observed at low speed, in particular at permeabilities below 10 to the -9th/squared cm. The present experimental and theoretical studies address questions of slip effects associated with long mean free paths of gas flow at room temperature. Data obtained are in good agreement, within data uncertainty, with a simplified asymptotic Knudsen equation proposed for porous plugs on the basis of Knudsen's classical flow equation for long mean free paths.

  12. Double-layered cell transfer technology for bone regeneration.

    PubMed

    Akazawa, Keiko; Iwasaki, Kengo; Nagata, Mizuki; Yokoyama, Naoki; Ayame, Hirohito; Yamaki, Kazumasa; Tanaka, Yuichi; Honda, Izumi; Morioka, Chikako; Kimura, Tsuyoshi; Komaki, Motohiro; Kishida, Akio; Izumi, Yuichi; Morita, Ikuo

    2016-01-01

    For cell-based medicine, to mimic in vivo cellular localization, various tissue engineering approaches have been studied to obtain a desirable arrangement of cells on scaffold materials. We have developed a novel method of cell manipulation called "cell transfer technology", enabling the transfer of cultured cells onto scaffold materials, and controlling cell topology. Here we show that using this technique, two different cell types can be transferred onto a scaffold surface as stable double layers or in patterned arrangements. Various combinations of adherent cells were transferred to a scaffold, amniotic membrane, in overlapping bilayers (double-layered cell transfer), and transferred cells showed stability upon deformations of the material including folding and trimming. Transplantation of mesenchymal stem cells from periodontal ligaments (PDLSC) and osteoblasts, using double-layered cell transfer significantly enhanced bone formation, when compared to single cell type transplantation. Our findings suggest that this double-layer cell transfer is useful to produce a cell transplantation material that can bear two cell layers. Moreover, the transplantation of an amniotic membrane with PDLSCs/osteoblasts by cell transfer technology has therapeutic potential for bone defects. We conclude that cell transfer technology provides a novel and unique cell transplantation method for bone regeneration. PMID:27624174

  13. Thermodynamic activity measurements of U-Zr alloys by knudsen effusion mass spectrometry

    NASA Astrophysics Data System (ADS)

    Kanno, Masayoshi; Yamawaki, Michio; Koyama, Tadafumi; Morioka, Nobuo

    1988-06-01

    Vaporization of a series of U-Zr alloys, a fundamental subsystem of the promising metallic fuel U-Pu-Zr, was studied by using a tantalum Knudsen cell coupled with a mass spectrometer in the temperature range 1700-2060 K. Thermodynamic activities, partial molar Gibbs free energies and integral molar Gibbs free energies of mixing were calculated from the partial vapor pressures of uranium over these alloys. The activities of uranium exhibit negative deviations from ideality, especially in the uranium-rich composition region. Both the solidus and liquidus lines for this system estimated from the activities show negative deviations from the tentative phase diagram previously reported.

  14. Measuring Thermodynamic Properties of Metals and Alloys With Knudsen Effusion Mass Spectrometry

    NASA Technical Reports Server (NTRS)

    Copland, Evan H.; Jacobson, Nathan S.

    2010-01-01

    This report reviews Knudsen effusion mass spectrometry (KEMS) as it relates to thermodynamic measurements of metals and alloys. First, general aspects are reviewed, with emphasis on the Knudsen-cell vapor source and molecular beam formation, and mass spectrometry issues germane to this type of instrument are discussed briefly. The relationship between the vapor pressure inside the effusion cell and the measured ion intensity is the key to KEMS and is derived in detail. Then common methods used to determine thermodynamic quantities with KEMS are discussed. Enthalpies of vaporization, the fundamental measurement, are determined from the variation of relative partial pressure with temperature using the second-law method or by calculating a free energy of formation and subtracting the entropy contribution using the third-law method. For single-cell KEMS instruments, measurements can be used to determine the partial Gibbs free energy if the sensitivity factor remains constant over multiple experiments. The ion-current ratio method and dimer-monomer method are also viable in some systems. For a multiple-cell KEMS instrument, activities are obtained by direct comparison with a suitable component reference state or a secondary standard. Internal checks for correct instrument operation and general procedural guidelines also are discussed. Finally, general comments are made about future directions in measuring alloy thermodynamics with KEMS.

  15. Double Stem Cell Transplant May Help Fight a Childhood Cancer

    MedlinePlus

    ... treatments might give survival edge to kids with neuroblastoma, researcher says To use the sharing features on ... The double stem cell transplant allows children with neuroblastoma to withstand two rounds of chemotherapy rather than ...

  16. Direct-drive DT implosions with Knudsen number variations

    NASA Astrophysics Data System (ADS)

    Kim, Y.; Herrmann, H. W.; Hoffman, N. M.; Schmitt, M. J.; Bradley, P. A.; Gales, S.; Horsfield, C. J.; Rubery, M.; Leatherland, A.; Gatu Johnson, M.; Frenje, J. A.; Glebov, V. Yu

    2016-05-01

    Direct-drive implosions of DT-filled plastic-shells have been conducted at the Omega laser facility, measuring nuclear yields while varying Knudsen numbers (i.e., the ratio of mean free path of fusing ions to the length of fuel region) by adjusting both shell thickness (e.g., 7.5, 15, 20, 30 μm) and fill pressure (e.g., 2, 5, 15 atm). The fusion reactivity reduction model showed a stronger effect on yield as the Knudsen number increases (or the shell thickness decreases). The Reduced-Ion-Kinetic (RIK) simulation which includes both fusion reactivity reduction and mix model was necessary to provide a better match between the observed neutron yields and those simulated.

  17. Double-sided solar-cell package

    NASA Technical Reports Server (NTRS)

    Shelpuk, B.

    1978-01-01

    Cost-effective solar-cell package is proposed for development of practical solar-cell system. Since cells are enclosed in inexpensive plastic tubes, forced-air-cooling loop is proposed to maintain cell temperature at adequate levels. Loop must include desiccant to remove moisture from hermetic enclosures to prevent cell corrosion.

  18. DSMC Simulation of Microstructure Actuation by Knudsen Thermal Force

    NASA Astrophysics Data System (ADS)

    Pikus, Aaron; Sebastiao, Israel; Strongrich, Andrew; Alexeenko, Alina

    2015-11-01

    Compact, low-power and highly accurate vacuum sensors are needed for emerging applications such as high-altitude communication platforms, small satellites and in-vacuum manufacturing processes. A novel MEMS-based pressure and gas sensor - Microelectromechanical In-plane Knudsen Radiometric Actuator (MIKRA) - has been developed at Purdue. MIKRA is based on Knudsen thermal force generated by rarefied flow driven by thermal gradients within the microstructure. The goal of this work is to model the rarefied gas flow in the MIKRA sensor to validate the numerical modeling of rarefied thermally-driven flows and gain insights for sensor design. The Direct Simulation Monte Carlo (DSMC) solver SPARTA is employed to numerically calculate the distribution of the flowfield and surface properties. The resulting forces on the colder shuttle beam are calculated and compared to the available experimental data as well as other numerical solvers. Both DSMC and experimental results suggest that the maximum forces occur at a Knudsen number of approximately 1. The streamlines indicate the presence of two small vortexes between the heated beam and the colder shuttle beam, and a larger one above these two beams. The DSMCsimulations, validated by experimental measurements, help understand the unique flow behaviors encountered in rarefied thermally-driven flows. NSF CBET grant #1055453.

  19. Vaporization Studies of Olivine via Knudsen Effusion Mass Spectrometry

    NASA Technical Reports Server (NTRS)

    Costa, G. C. C.; Jacobson, N. S.

    2014-01-01

    Olivine is the major mineral in the Earth's upper mantle occurring predominantly in igneous rocks and has been identified in meteorites, asteroids, the Moon and Mars. Among many other important applications in planetary and materials sciences, the thermodynamic properties of vapor species from olivine are crucial as input parameters in computational modelling of the atmospheres of hot, rocky exoplanets (lava planets). There are several weight loss studies of olivine vaporization in the literature and one Knudsen Effusion Mass Spectrometry (KEMS) study. In this study, we examine a forsterite-rich olivine (93% forsterite and 7% fayalite, Fo93Fa7) with KEMS to further understand its vaporization and thermodynamic properties.

  20. Modeling Ablation of Fibrous Materials from Bulk to Knudsen Regime

    NASA Technical Reports Server (NTRS)

    Lachaud, Jean; Mansour, Nagi N.

    2008-01-01

    Material-environment interactions are analyzed at microscopic scale to explain the lower than expected density observed by post-flight analysis of the char layer on the Stardust shield. Mass transfer, ablation (oxidation), and surface recession of fibrous material is simulated in 3D using a Monte-Carlo simulation tool. Ablation is found to occur either at the surface or in volume depending on Knudsen and Thiele number values. This study supports the idea of volume ablation followed by possible carbon fiber spallation that may explain post-flight analyses.

  1. Design and experimental validation of a compact collimated Knudsen source.

    PubMed

    Wouters, Steinar H W; Ten Haaf, Gijs; Mutsaers, Peter H A; Vredenbregt, Edgar J D

    2016-08-01

    In this paper, the design and performance of a collimated Knudsen source, which has the benefit of a simple design over recirculating sources, is discussed. Measurements of the flux, transverse velocity distribution, and brightness of the resulting rubidium beam at different source temperatures were conducted to evaluate the performance. The scaling of the flux and brightness with the source temperature follows the theoretical predictions. The transverse velocity distribution in the transparent operation regime also agrees with the simulated data. The source was tested up to a temperature of 433 K and was able to produce a flux in excess of 10(13) s(-1). PMID:27587111

  2. A bicontinuous double gyroid hybrid solar cell.

    PubMed

    Crossland, Edward J W; Kamperman, Marleen; Nedelcu, Mihaela; Ducati, Caterina; Wiesner, Ulrich; Smilgies, Detlef-M; Toombes, Gilman E S; Hillmyer, Marc A; Ludwigs, Sabine; Steiner, Ullrich; Snaith, Henry J

    2009-08-01

    We report the first successful application of an ordered bicontinuous gyroid semiconducting network in a hybrid bulk heterojunction solar cell. The freestanding gyroid network is fabricated by electrochemical deposition into the 10 nm wide voided channels of a self-assembled, selectively degradable block copolymer film. The highly ordered pore structure is ideal for uniform infiltration of an organic hole transporting material, and solid-state dye-sensitized solar cells only 400 nm thick exhibit up to 1.7% power conversion efficiency. This patterning technique can be readily extended to other promising heterojunction systems and is a major step toward realizing the full potential of self-assembly in the next generation of device technologies. PMID:19007289

  3. Microstructure actuation and gas sensing by the Knudsen thermal force

    SciTech Connect

    Strongrich, Andrew; Alexeenko, Alina

    2015-11-09

    The generation of forces and moments on structures immersed in rarefied non-isothermal gas flows has received limited practical implementation since first being discovered over a century ago. The formation of significant thermal stresses requires both large thermal gradients and characteristic dimensions which are comparable to the gas molecular mean free path. For macroscopic geometries, this necessitates impractically high temperatures and very low pressures. At the microscale, however, these conditions are easily achieved, allowing the effects to be exploited, namely, for gas-property sensing and microstructure actuation. In this letter, we introduce and experimentally evaluate performance of a microelectromechanical in-plane Knudsen radiometric actuator, a self-contained device having Knudsen thermal force generation, sensing, and tuning mechanisms integrated onto the same platform. Sensitivity to ambient pressure, temperature gradient, as well as gas composition is demonstrated. Results are presented in terms of a non-dimensional force coefficient, allowing measurements to be directly compared to the previous experimental and computational data on out-of-plane cantilevered configurations.

  4. On the role of the Knudsen layer in rapid granular flows

    SciTech Connect

    Galvin, J.E.; Hrenya, C.M.; Wildman, R.D.

    2007-08-01

    A combination of molecular dynamics simulations, theoretical predictions and previous experiments are used in a two-part study to determine the role of the Knudsen layer in rapid granular flows. First, a robust criterion for the identification of the thickness of the Knudsen layer is established: a rapid deterioration in Navier–Stokes order prediction of the heat flux is found to occur in the Knudsen layer. For (experimental) systems in which heat flux measurements are not easily obtained, a rule-of-thumb for estimating the Knudsen layer thickness follows, namely that such effects are evident within 2.5 (local) mean free paths of a given boundary. Secondly, comparisons of simulation and experimental data with Navier–Stokes order theory are used to provide a measure as to when Knudsen-layer effects become non-negligible. Specifically, predictions that do not account for the presence of a Knudsen layer appear reliable for Knudsen layers collectively composing up to 20% of the domain, whereas deterioration of such predictions becomes apparent when the domain is fully comprised of the Knudsen layer.

  5. On the role of the Knudsen layer in rapid granular flows

    NASA Astrophysics Data System (ADS)

    Galvin, J. E.; Hrenya, C. M.; Wildman, R. D.

    2007-08-01

    A combination of molecular-dynamics simulations, theoretical predictions, and previous experiments are used in a two-part study to determine the role of the Knudsen layer in rapid granular flows. First, a robust criterion for the identification of the thickness of the Knudsen layer is established: a rapid deterioration in Navier-Stokes-order prediction of the heat flux is found to occur in the Knudsen layer. For (experimental) systems in which heat flux measurements are not easily obtained, a rule-of-thumb for estimating the Knudsen layer thickness follows, namely that such effects are evident within 2.5 (local) mean free paths of a given boundary. Second, comparisons of simulation and experimental data with Navier-Stokes order theory are used to provide a measure as to when Knudsen layer effects become non-negligible. Specifically, predictions that do not account for the presence of a Knudsen layer appear reliable for Knudsen layers collectively composing up to 20% of the domain, whereas deterioration of such predictions becomes apparent when the domain is fully comprised of the Knudsen layer.

  6. Upscaling momentum and mass transport under Knudsen and binary diffusion gas slip conditions

    NASA Astrophysics Data System (ADS)

    Valdes-Parada, F. J.; Lasseux, D.

    2015-12-01

    Modeling of gas phase flow in porous media is relevant as it is present in a wide variety of applications ranging from nanofluidic systems to subsurface contaminant transport. In this work, we derive a macroscopic model to study slightly compressible gas flow in porous media for conditions in which the tangential fluid velocity undergoes a slip at the solid interface due to Knudsen effects and to mass diffusion in binary conditions. To this end, we use the method of volume averaging to derive the governing equations at the Darcy scale for both mass and momentum transport. The momentum transport model consists on a modification to Darcy's law due to mass dispersion and to total density gradients. For mass transport, the resulting model is the conventional convection-dispersion equation with two correction terms, one affecting convective transport and the second one affecting mass dispersion due to gas compressibility. The macroscopic model reduces to the one reported by Altevogt et al. (2003) for the case in which gas slip is only due to a concentration gradient and to the one by Lasseux et al. (2014) under Knudsen slip conditions. The model is written in terms of effective-medium coefficients that can be predicted from solving the associated closure problems in representative unit cells. For conditions in which the Péclet number is much greater than one and when the Knudsen number is not exceedingly small compared to the unity, our computations show that the predictions of the longitudinal dispersion may reach an error as high as 60% compared to the predictions obtained by ignoring gas slip. Altevogt A.S., Rolston D.E., Whitaker S. New equations for binary gas transport in porous media, Part 1: equation development. Advances in Water Resources, Vol. 26, 695-715, 2003. Lasseux D., Valdés-Parada F.J., Ochoa-Tapia J.A., Goyeau B. A macroscopic model for slightly compressible gas slip-flow in homogeneous porous media. Physics of Fluids, Vol. 26, 053102, 2014.

  7. Revised Knudsen-layer reduction of fusion reactivity

    SciTech Connect

    Albright, B. J.; Molvig, Kim; Huang, C.-K.; Simakov, A. N.; Dodd, E. S.; Hoffman, N. M.; Kagan, G.; Schmit, P. F.

    2013-12-15

    Recent work by Molvig et al. [Phys. Rev. Lett. 109, 095001 (2012)] examined how fusion reactivity may be reduced from losses of fast ions in finite assemblies of fuel. In this paper, this problem is revisited with the addition of an asymptotic boundary-layer treatment of ion kinetic losses. This boundary solution, reminiscent of the classical Milne problem from linear transport theory, obtains a free-streaming limit of fast ion losses near the boundary, where the diffusion approximation is invalid. Thermonuclear reaction rates have been obtained for the ion distribution functions predicted by this improved model. It is found that while Molvig's “Knudsen distribution function” bounds from above the magnitude of the reactivity reduction, this more accurate treatment leads to less dramatic losses of tail ions and associated reduction of thermonuclear reaction rates for finite fuel volumes.

  8. Phonon Knudsen flow in GaAs/AlAs superlattices

    SciTech Connect

    Hyldgaard, P.; Mahan, G.D. |

    1995-09-01

    The measured in-plane thermal conductivity, {delta}{sub SL} of GaAs/AlAs superlattices with even moderate layer thicknesses are significantly smaller than the weighted average, {delta}{sub l} = 67 W/Km, of the bulk GaAs and AlAs conductivities. One expects a suppression of the thermal conductivity to that of an actual Al{sub 0.5}Ga{sub 0.5}As alloy when the thickness of the GaAs and AlAs layers approaches that of a single monolayer. However, the observed superlattice thermal conductivity remains suppressed even at layer thickness {approx_gt} 10 nm. The low thermal conductivities, and very high mobilities, make n-doped GaAs/AlAs superlattices attractive possibilities for thermoelectric devices. With Molecular-Beam-Epitaxial grown GaAs/AlAs superlattices one can expect the individual GaAs and AlAs layers to be extremely clean. Defect and/or alloy scattering is limited to be near the heterostructure interfaces. The authors estimate the room-temperature phonon mean-free-path to be 42 (22) nm for the longitudinal (transverse) mode and thus comparable to or smaller than the layer thicknesses. Thus they expect an important phonon scattering at the interfaces. They study this phonon scattering at the superlattice interfaces assuming a Knudsen flow characterized by diffusive scattering. The solid curve in the figure shows the Knudsen-flow theory estimated for the superlattice thermal conductivity which shows a significant reduction when the layer thickness is shorter than the estimated phonon mean free paths.

  9. Wearable double-twisted fibrous perovskite solar cell.

    PubMed

    Li, Ru; Xiang, Xi; Tong, Xiao; Zou, Jingyun; Li, Qingwen

    2015-07-01

    Wearable double-twisted fibrous perovskite solar cells are developed based on flexible carbon nanotube fiber electrodes, which exhibit a maximum power conversion efficiency of 3.03% and bending stability larger than 1000 cycles, and maintain 89% efficiency after 96 h in ambient conditions if sealed by a transparent polymer layer. The obtained superior performance can shed light on future self-powering e-textiles. PMID:25989248

  10. A wall-function approach to incorporating Knudsen-layer effects in gas micro flow simulations.

    SciTech Connect

    Gallis, Michail A.; Lockerby, Duncan A.; Reese, Jason M.

    2004-07-01

    For gas flows in microfluidic configurations, the Knudsen layer close to the wall can comprise a substantial part of the entire flowfield and has a major effect on quantities such as the mass flow rate through micro devices. The Knudsen layer itself is characterized by a highly nonlinear relationship between the viscous stress and the strain rate of the gas, so even if the Navier-Stokes equations can be used to describe the core gas flow they are certainly inappropriate for the Knudsen layer itself. In this paper we propose a 'wall-function' model for the stress/strain rate relations in the Knudsen layer. The constitutive structure of the Knudsen layer has been derived from results from kinetic theory for isothermal shear flow over a planar surface. We investigate the ability of this simplified model to predict Knudsen-layer effects in a variety of configurations. We further propose a semi-empirical Knudsen-number correction to this wall function, based on high-accuracy DSMC results, to extend the predictive capabilities of the model to greater degrees of rarefaction.

  11. High-temperature mass spectrometry - Vaporization of group 4-B metal carbides. [using Knudsen effusion

    NASA Technical Reports Server (NTRS)

    Stearns, C. A.; Kohl, F. J.

    1974-01-01

    The high temperature vaporization of the metal-carbon systems TiC, ZrC, HfC, and ThC was studied by the Knudsen effusion - mass spectrometric method. For each system the metal dicarbide and tetracarbide molecular species were identified in the gas phase. Relative ion currents of the carbides and metals were measured as a function of temperature. Second- and third-law methods were used to determine enthalpies. Maximum values were established for the dissociation energies of the metal monocarbide molecules TiC, ZrC, HfC, and ThC. Thermodynamic functions used in the calculations are discussed in terms of assumed molecular structures and electronic contributions to the partition functions. The trends shown by the dissociation energies of the carbides of Group 4B are compared with those of neighboring groups and discussed in relation to the corresponding oxides and chemical bonding. The high temperature molecular beam inlet system and double focusing mass spectrometer are described.

  12. KNUDSEN CELL REACTOR FOR CATALYST RESEARCH RELATED TO HYDROGEN TECHNOLOGIES

    EPA Science Inventory

    Hydrogen has been identified as a viable sustainable alternative to fossil fuels. Hydrogen as an energy source is ecologically feasible, socially desirable, and with continued research and development promises to become economically viable. The faculty advisors listed...

  13. Solution-processed crystalline silicon double-heterojunction solar cells

    NASA Astrophysics Data System (ADS)

    Devkota, Ramesh; Liu, Qiming; Ohki, Tatsuya; Hossain, Jaker; Ueno, Keiji; Shirai, Hajime

    2016-02-01

    Crystalline silicon double-heterojunction solar cells were fabricated using Si/organic and Si/Cs2CO3 heterojunctions. The front heterojunction is formed by spin-coating conductive polymer poly(3,4-ethyenedioxythiophene):poly(styrene sulfonate) (PEDOT:PSS) on n-type Czochralski (CZ) (100) silicon, which separates the photogenerated carriers and blocks the electron dark current while allowing the photocurrent to pass through. The rear heterojunction, formed by spin-coating Cs2CO3 and polyethylenimine (PEI) dissolved in 2-ethoxyethanol and Al metal evaporation, functions as a back surface field that reduces the hole dark current while allowing the electron photocurrent to pass through. The double-heterojunction device showed a power conversion efficiency of 12.7% under AM1.5G simulated solar light exposure.

  14. Thermoresponsive double network micropillared hydrogels for controlled cell release.

    PubMed

    Fei, Ruochong; Hou, Huijie; Munoz-Pinto, Dany; Han, Arum; Hahn, Mariah S; Grunlan, Melissa A

    2014-09-01

    Thermoresponsive poly(N-isopropylacrylamide) hydrogels (PNIPAAm) have been widely used for controlled cell detachment. In this study, cell release is enhanced via deswelling with a two-pronged approach combining a double network (DN) design and micropatterning. PNIPAAm hydrogels are prepared as DNs comprised of a tightly crosslinked 1st network and a loosely crosslinked 2nd network. Moreover, the PNIPAAm DN hydrogels are prepared as both planar 1.5 mm-thick slabs as well as micropillar arrays (≈200 μm pillar diameter). Compared to the corresponding conventional single network (SN) hydrogels, DN hydrogels exhibit enhanced thermosensitivity and cell release efficiency, particularly for the micropillar arrays. PMID:24956117

  15. Knudsen Reactivity Reduction: Kinetic Theory of Diffusion Process

    NASA Astrophysics Data System (ADS)

    Nelson, Eric; Dodd, Evan; Molvig, Kim; Albright, Brian; Hoffman, Nelson; Zimmerman, George; Williams, Ed

    2012-10-01

    Previous work that found significant fusion reactivity reduction due to Knudsen layer losses [1], utilized a twice simplified treatment of the loss process that first went to the diffusion limit of the transport and then replaced the spatial kinetic diffusion operator by a local loss process. The derivation of kinetic diffusion utilized a stochastic differential equation technique to show that convection in combination with pitch-angle scattering yields spatial diffusion asymptotically over long time and spatial intervals. The same technique can be extended to include the independent energy scattering stochastic process. For the linear Fokker-Planck equation that governs the tail ions this gives a very efficient (particle like) numerical technique that can solve the complete ion tail problem in the three phase space dimensions of pitch-angle, energy, and spatial coordinate. The method allows inclusion of a temperature gradient and specified ambipolar electric fields. We present simulation results of the depleted tail distributions and fusion reactivities, and compare with the predictions of the simple local loss method.[4pt] [1] Kim Molvig, Nelson N. Hoffman, Brian J. Albright, Eric M. Nelson, and, Robert J. Webster (submitted to Physical Review Letters, 2012)

  16. Thermodynamic limit of bifacial double-junction tandem solar cells

    NASA Astrophysics Data System (ADS)

    Ryyan Khan, M.; Alam, Muhammad A.

    2015-11-01

    A traditional single-junction solar panel cannot harness ground-scattered light (albedo reflectance, RA ), and also suffers from the fundamental sub-band-gap and the thermalization losses. In this paper, we explain how a "bifacial tandem" panel would dramatically reduce these losses, with corresponding improvement in thermodynamic performance. Our study predicts (i) the optimum combination of the band-gaps, empirically given by Eg(t ) o p t≈Eg(b ) o p t(2 +RA)/3 +(1 -RA) and the (ii) corresponding optimum normalized output power given by ηT(op t ) *≈RA (2 ηSJ (o p t ) ) +(1 -RA ) ηDJ (o p t ) . Empirically, ηT(op t ) * interpolates between the thermodynamic efficiency limit of classical double-junction tandem cell ( ηDJ ) and twice that of a single-junction cell ( ηSJ ). We conclude by explaining how the fundamental loss mechanisms evolve with RA in a bifacial tandem cell.

  17. The MEMS Knudsen Compressor as a Vacuum Pump for Space Exploration Applications

    NASA Technical Reports Server (NTRS)

    Vargo, S. E.; Muntz, E. P.; Tang, W. C.

    2000-01-01

    Several lander, probe and rover missions currently under study at the Jet Propulsion Laboratory (JPL) and especially in the Microdevices Laboratory (MDL) Center for Space Microelectronics Technology, focus on utilizing microelectromechanical systems (MEMS) based instruments for science data gathering. These small instruments and NASA's commitment to "faster, better, cheaper" type missions has brought about the need for novel approaches to satisfying mission requirements. Existing in-situ instrument systems clearly lack novel and integrated methods for satisfying their vacuum needs. One attractive candidate for a MEMS vacuum pump is the Knudsen Compressor, which operates based on thermal transpiration. Thermal transpiration describes gas flows induced by temperature differences maintained across orifices, porous membranes or capillary tubes under rarefied conditions. This device has two overwhelmingly attractive features as a MEMS vacuum pump - no moving parts and no fluids. An initial estimate of a Knudsen Compressor's pumping power requirements for a surface atmospheric sampling task on Mars is less than 80 mW, significantly below than alternative pumps. Due to the relatively low energy use for this task and the applicability of the Knudsen Compressor to other applications, the development of a Knudsen Compressor utilizing MEMS fabrication techniques has been initiated. This paper discusses the initial fabrication of a single-stage MEMS Knudsen Compressor vacuum pump, provides performance criteria such as pumping speed, size, energy use and ultimate pressure and details vacuum pump applications in several MDL related in-situ instruments.

  18. Clinical Significance of “Double-hit” and “Double-protein” expression in Primary Gastric B-cell Lymphomas

    PubMed Central

    He, Miaoxia; Chen, Keting; Li, Suhong; Zhang, Shimin; Zheng, Jianming; Hu, Xiaoxia; Gao, Lei; Chen, Jie; Song, Xianmin; Zhang, Weiping; Wang, Jianmin; Yang, Jianmin

    2016-01-01

    BACKGROUND AND AIMS: Primary gastric B-cell lymphoma is the second most common malignancy of the stomach. There are many controversial issues about its diagnosis, treatment and clinical management. “Double-hit” and “double-protein” involving gene rearrangement and protein expression of c-Myc and bcl2/bcl6 are the most used terms to describe DLBCL poor prognostic factors in recent years. However, very little is known about the role of these prognostic factors in primary gastric B-cell lymphomas. This study aims to obtain a molecular pathology prognostic model of gastric B-cell lymphoma for clinical stratified management by evaluating how the “double-hit” and “double-protein” in tumor cells as well as microenvironmental reaction of tumor stromal tissue affect clinical outcome in primary gastric B-cell lymphomas. METHODS: Data and tissues of 188 cases diagnosed with gastric B-cell lymphomas were used in this study. Tumor tissue microarray (TMA) of formalin fixed and paraffin embedded (FFPE) tissues was constructed for fluorescence in situ hybridization (FISH) and immunohistochemistry (IHC) analysis with a serial of biomarkers containing MYC, BCL2, BCL6, CD31, SPARC, CD10, MUM1 and Ki-67. Modeled period analysis was used to estimate 3-year and 5-year overall survival (OS) and disease-free survival (DFS) distributions. RESULTS: There was no definite “double-hit” case though the gene rearrangement of c-Myc (5.9%), bcl2 (0.1%) and bcl6 (7.4%) was found in gastric B-cell lymphomas. The gene amplification or copy gains of c-Myc (10.1%), bcl-2 (17.0%) and bcl-6 (0.9%) were present in these lymphomas. There were 12 cases of the lymphomas with the “double-protein” expression of MYC and BCL2/BCL6. All patients with “double-protein” gastric B-cell lymphomas had poor outcome compared with those without. More importantly, “MYC-BCL2-BCL6” negative group of gastric B-cell lymphoma patients had favorable clinical outcome regardless clinical stage

  19. Development of a new correlation to calculate permeability for flows with high Knudsen number

    NASA Astrophysics Data System (ADS)

    Esmaeil, Dehdashti

    2016-02-01

    Flows with high Knudsen number play a prominent role in many engineering applications. The present study is an effort toward the simulation of flow with high Knudsen number using modified lattice Boltzmann method (LBM) through a porous medium in a channel. The effect of collision between molecules and solid walls, which is required to accurately simulate transition flow regime, is taken into account using a modified relaxation time. Slip velocity on the wall, which is another significant difficulty in simulating transition flow regime, is captured using the slip reflection boundary condition (SRBC). The geometry of porous medium is considered as in-line and staggered. The results are in good agreement with previous works. A new correlation is obtained between permeability, Knudsen number and porosity for flows in transition flow regimes.

  20. Increasing cell culture population doublings for long-term growth of finite life span human cell cultures

    SciTech Connect

    Stampfer, Martha R; Garbe, James C

    2015-02-24

    Cell culture media formulations for culturing human epithelial cells are herein described. Also described are methods of increasing population doublings in a cell culture of finite life span human epithelial cells and prolonging the life span of human cell cultures. Using the cell culture media disclosed alone and in combination with addition to the cell culture of a compound associated with anti-stress activity achieves extended growth of pre-stasis cells and increased population doublings and life span in human epithelial cell cultures.

  1. Increasing cell culture population doublings for long-term growth of finite life span human cell cultures

    DOEpatents

    Stampfer, Martha R.; Garbe, James C.

    2016-06-28

    Cell culture media formulations for culturing human epithelial cells are herein described. Also described are methods of increasing population doublings in a cell culture of finite life span human epithelial cells and prolonging the life span of human cell cultures. Using the cell culture media disclosed alone and in combination with addition to the cell culture of a compound associated with anti-stress activity achieves extended growth of pre-stasis cells and increased population doublings and life span in human epithelial cell cultures.

  2. Development and preliminary evaluation of a double-cell ozonesonde

    NASA Astrophysics Data System (ADS)

    Zhang, Jinqiang; Xuan, Yuejian; Yan, Xiaolu; Liu, Mingyuan; Tian, Hongmin; Xia, Xiang'ao; Pang, Li; Zheng, Xiangdong

    2014-07-01

    Ozonesondes are widely used to obtain ozone concentration profiles from the surface to the upper atmosphere. A kind of double-cell ozonesonde has been developed at the Institute of Atmospheric Physics (IAP), Chinese Academy of Sciences (named the "IAP ozonesonde") based on previous experience over the past 20 years of developing the singlecell GPSO3 ozonesonde. The IAP ozonesonde is of the Electrochemical Concentration Cell (ECC) type. A detailed description of the IAP ozonesonde is firstly provided in the present paper, followed by a presentation of results from a series of launches carried out to evaluate its performance. The analysis involved comparing its observations with measurements from the GPSO3 and ECC ozonesondes (Model type ENSCI-Z) as well as a Brewer spectrophotometer. The results showed that the IAP ozonesonde is a vast improvement over the GPSO3 ozonesonde, able to capture vertical ozone structures very well and in good agreement with ECC ozonesonde measurements. The average difference in the ozone partial pressure between the IAP and ECC ozonesondes was 0.3 mPa from the surface to 2.5 km, close to zero from 2.5 to 9 km and generally less than 1 mPa for layers higher than 9 km. The apparent deviation is likely caused by a decreasing pump flow rate in the IAP ozonesonde which needs further improvement. The total ozone amounts measured by the IAP ozonesonde profiles were highly comparable with the Brewer data with a relative difference of 6%. The development of the IAP ozonesonde and its strong performance will surely accelerate the process of conventional observations of ozone profiles over China in the near future as well as provide more data for ozone research in general.

  3. Measuring red blood cell aggregation forces using double optical tweezers.

    PubMed

    Fernandes, Heloise P; Fontes, Adriana; Thomaz, André; Castro, Vagner; Cesar, Carlos L; Barjas-Castro, Maria L

    2013-04-01

    Classic immunohematology approaches, based on agglutination techniques, have been used in manual and automated immunohematology laboratory routines. Red blood cell (RBC) agglutination depends on intermolecular attractive forces (hydrophobic bonds, Van der Walls, electrostatic forces and hydrogen bonds) and repulsive interactions (zeta potential). The aim of this study was to measure the force involved in RBC aggregation using double optical tweezers, in normal serum, in the presence of erythrocyte antibodies and associated to agglutination potentiator solutions (Dextran, low ionic strength solution [LISS] and enzymes). The optical tweezers consisted of a neodymium:yattrium aluminium garnet (Nd:YAG) laser beam focused through a microscope equipped with a minicam, which registered the trapped cell image in a computer where they could be analyzed using a software. For measuring RBC aggregation, a silica bead attached to RBCs was trapped and the force needed to slide one RBC over the other, as a function of the velocities, was determined. The median of the RBC aggregation force measured in normal serum (control) was 1 × 10(-3) (0.1-2.5) poise.cm. The samples analyzed with anti-D showed 2 × 10(-3) (1.0-4.0) poise.cm (p < 0.001). RBC diluted in potentiator solutions (Dextran 0.15%, Bromelain and LISS) in the absence of erythrocyte antibodies, did not present agglutination. High adherence was observed when RBCs were treated with papain. Results are in agreement with the imunohematological routine, in which non-specific results are not observed when using LISS, Dextran and Bromelain. Nevertheless, false positive results are frequently observed in manual and automated microplate analyzer using papain enzyme. The methodology proposed is simple and could provide specific information with the possibility of meansuration regarding RBC interaction. PMID:23402665

  4. High-order lattice Boltzmann models for wall-bounded flows at finite Knudsen numbers

    NASA Astrophysics Data System (ADS)

    Feuchter, C.; Schleifenbaum, W.

    2016-07-01

    We analyze a large number of high-order discrete velocity models for solving the Boltzmann-Bhatnagar-Gross-Krook equation for finite Knudsen number flows. Using the Chapman-Enskog formalism, we prove for isothermal flows a relation identifying the resolved flow regimes for low Mach numbers. Although high-order lattice Boltzmann models recover flow regimes beyond the Navier-Stokes level, we observe for several models significant deviations from reference results. We found this to be caused by their inability to recover the Maxwell boundary condition exactly. By using supplementary conditions for the gas-surface interaction it is shown how to systematically generate discrete velocity models of any order with the inherent ability to fulfill the diffuse Maxwell boundary condition accurately. Both high-order quadratures and an exact representation of the boundary condition turn out to be crucial for achieving reliable results. For Poiseuille flow, we can reproduce the mass flow and slip velocity up to the Knudsen number of 1. Moreover, for small Knudsen numbers, the Knudsen layer behavior is recovered.

  5. The usefulness of higher-order constitutive relations for describing the Knudsen layer.

    SciTech Connect

    Gallis, Michail A.; Lockerby, Duncan A.; Reese, Jason M.

    2005-03-01

    The Knudsen layer is an important rarefaction phenomenon in gas flows in and around microdevices. Its accurate and efficient modeling is of critical importance in the design of such systems and in predicting their performance. In this paper we investigate the potential that higher-order continuum equations may have to model the Knudsen layer, and compare their predictions to high-accuracy DSMC (direct simulation Monte Carlo) data, as well as a standard result from kinetic theory. We find that, for a benchmark case, the most common higher-order continuum equation sets (Grad's 13 moment, Burnett, and super-Burnett equations) cannot capture the Knudsen layer. Variants of these equation families have, however, been proposed and some of them can qualitatively describe the Knudsen layer structure. To make quantitative comparisons, we obtain additional boundary conditions (needed for unique solutions to the higher-order equations) from kinetic theory. However, we find the quantitative agreement with kinetic theory and DSMC data is only slight.

  6. Modeling of the Ablation of Fibrous Materials in the Knudsen Regime

    NASA Technical Reports Server (NTRS)

    Lachaud, J.; Mansour, N. N.

    2008-01-01

    During atmospheric entry of planetary probes, the thermal protection system (TPS) of the probe is exposed to high temperatures under low pressures. In these conditions, carbonous TPS materials undergo gasification in the Knudsen regime leading to mass loss and wall recession called ablation. This work aims to improve the understanding of materiaVenvironment interactions through a study of the coupling between carbon dioxide transport in the Knudsen regime, heterogeneous oxidation of carbon, and sutface recession. A 3D Monte-Carlo simulation tool is used for this study. The fibrous architecture of the materiils, consisting of high porosity random array of carbon fibers, is numerically reproduced on a 3D Cartesian grid. Mass transport in the Knudsen regime from the boundary layer to the surface, and inside this porous material is simulated by random walk. A reaction probability is used to simulate the heterogeneous oxidation reaction. The surface recession is followed by front tracking using a simplified marching cube approach. The output data of the simulations are ablation velocity and dynamic evolution of the material porosity. A parametric study is carried out to analyze the material behavior as a function of Knudsen number for the porous media (length of the mean free path compared to the mean pore diameter) and the intrinsic reactivity of the carbon fibers. The results enable extrapolation of laboratory experimental data to actual entry conditions.

  7. Studying the volatility of pyrazolone complexes of rare-earth elements by means of Knudsen effusion

    NASA Astrophysics Data System (ADS)

    Lazarev, N. M.; Petrov, B. I.; Bochkarev, L. N.; Safronova, A. V.; Abakumov, G. A.; Arapova, A. V.; Bessonova, Yu. A.

    2014-08-01

    The temperature dependences of the pressure of saturated vapor of pyrazolone complexes of rare-earth elements Ln(PMIP)3 (where Ln = Y, Ho, Er, Tm, Lu; PMIP = 1-phenyl-3-methyl-4-isobutyryl-5-pyrazolone) are studied via Knudsen effusion, and the enthalpy of their sublimation is determined. Mass spectra and differential scanning calorimetry data are obtained.

  8. Discrete unified gas kinetic scheme for all Knudsen number flows. II. Thermal compressible case.

    PubMed

    Guo, Zhaoli; Wang, Ruijie; Xu, Kun

    2015-03-01

    This paper is a continuation of our work on the development of multiscale numerical scheme from low-speed isothermal flow to compressible flows at high Mach numbers. In our earlier work [Z. L. Guo et al., Phys. Rev. E 88, 033305 (2013)], a discrete unified gas kinetic scheme (DUGKS) was developed for low-speed flows in which the Mach number is small so that the flow is nearly incompressible. In the current work, we extend the scheme to compressible flows with the inclusion of thermal effect and shock discontinuity based on the gas kinetic Shakhov model. This method is an explicit finite-volume scheme with the coupling of particle transport and collision in the flux evaluation at a cell interface. As a result, the time step of the method is not limited by the particle collision time. With the variation of the ratio between the time step and particle collision time, the scheme is an asymptotic preserving (AP) method, where both the Chapman-Enskog expansion for the Navier-Stokes solution in the continuum regime and the free transport mechanism in the rarefied limit can be precisely recovered with a second-order accuracy in both space and time. The DUGKS is an idealized multiscale method for all Knudsen number flow simulations. A number of numerical tests, including the shock structure problem, the Sod tube problem in a whole range of degree of rarefaction, and the two-dimensional Riemann problem in both continuum and rarefied regimes, are performed to validate the scheme. Comparisons with the results of direct simulation Monte Carlo (DSMC) and other benchmark data demonstrate that the DUGKS is a reliable and efficient method for multiscale flow problems. PMID:25871252

  9. Discrete unified gas kinetic scheme for all Knudsen number flows. II. Thermal compressible case

    NASA Astrophysics Data System (ADS)

    Guo, Zhaoli; Wang, Ruijie; Xu, Kun

    2015-03-01

    This paper is a continuation of our work on the development of multiscale numerical scheme from low-speed isothermal flow to compressible flows at high Mach numbers. In our earlier work [Z. L. Guo et al., Phys. Rev. E 88, 033305 (2013), 10.1103/PhysRevE.88.033305], a discrete unified gas kinetic scheme (DUGKS) was developed for low-speed flows in which the Mach number is small so that the flow is nearly incompressible. In the current work, we extend the scheme to compressible flows with the inclusion of thermal effect and shock discontinuity based on the gas kinetic Shakhov model. This method is an explicit finite-volume scheme with the coupling of particle transport and collision in the flux evaluation at a cell interface. As a result, the time step of the method is not limited by the particle collision time. With the variation of the ratio between the time step and particle collision time, the scheme is an asymptotic preserving (AP) method, where both the Chapman-Enskog expansion for the Navier-Stokes solution in the continuum regime and the free transport mechanism in the rarefied limit can be precisely recovered with a second-order accuracy in both space and time. The DUGKS is an idealized multiscale method for all Knudsen number flow simulations. A number of numerical tests, including the shock structure problem, the Sod tube problem in a whole range of degree of rarefaction, and the two-dimensional Riemann problem in both continuum and rarefied regimes, are performed to validate the scheme. Comparisons with the results of direct simulation Monte Carlo (DSMC) and other benchmark data demonstrate that the DUGKS is a reliable and efficient method for multiscale flow problems.

  10. Transdifferentiation of human amniotic epithelial cells into acinar cells using a double-chamber system.

    PubMed

    Huang, Gui-Lin; Zhang, Ni-Ni; Wang, Jun-Sheng; Yao, Li; Zhao, Yu-Jie; Wang, Yu-Ying

    2012-08-01

    This study investigated the transdifferentiation of stem cells from human amnion tissue into functional acinar cells (ACs) using a co-culture system. Human amniotic epithelial cells (hAECs) were isolated from amnion tissue by mechanical mincing and enzymatic digestion. After primary culture, the phenotype of the cells was identified by flow cytometry (FCM) and immunocytochemical staining. hAECs were co-cultured with submandibular gland acinar cells of SD rats using a double-chamber system. The expression of α-amylase was determined by immunocytochemical method and fluorescent real-time quantitative reverse transcription polymerase chain reaction (RT-PCR) after induction for 1 and 2 weeks, respectively. Digestion with trypsin is an effective method for isolating hAECs from amnion tissue. These cells were positive for CD29 and CK19 and weakly positive for CD44 and α-amylase. Within 2 weeks, α-amylase in hAECs increased with induction time. The expression of α-amylase in hAECs was increased 3.38-fold after co-culturing for 1 week. This ratio increased to 6.6-fold, and these cells were positive for mucins, after co-culturing for 2 weeks. hAECs possess the potential to differentiate into ACs in vitro. They might be a stem cell resource for clinical applications of cell replacement therapy in salivary gland dysfunction diseases. PMID:22800093

  11. Eugenic metal-free sensitizers with double anchors for high performance dye-sensitized solar cells.

    PubMed

    Hung, Wei-I; Liao, You-Ya; Lee, Ting-Hui; Ting, Yu-Chien; Ni, Jen-Shyang; Kao, Wei-Siang; Lin, Jiann T; Wei, Tzu-Chien; Yen, Yung-Sheng

    2015-02-01

    A series of new phenothiazine-based dyes (HL5-HL7) with double acceptors/anchors have been synthesized and used as the sensitizers for highly efficient dye-sensitized solar cells (DSSCs). Among them, the HL7-based cell exhibits the best efficiency of 8.32% exceeding the N719-based cell (7.35%) by ∼13%. PMID:25555237

  12. High-efficiency double-heterostructure AlGaAs/GaAs solar cells

    NASA Technical Reports Server (NTRS)

    Gale, R. P.; Fan, John C. C.; Turner, G. W.; Chapman, R. L.

    1985-01-01

    Double-heterostructure solar cells have been fabricated from wafers prepared by using organometallic chemical vapor deposition to grow a p GaAs absorbing layer sandwiched between p(+) and n(+) AlGaAs layers. The best cell, which incorporates an abrupt AlGaAs/GaAs shallow heterojunction, exhibits a global AM1 one-sun conversion efficiency of 23 percent. The rate at which the open-circuit voltage decreases with increasing temperature is lower for the double-heterostructure cells than for GaAs shallow-homojunction cells.

  13. Arrayed architectures for multi-stage Si-micromachined high-flow Knudsen pumps

    NASA Astrophysics Data System (ADS)

    Qin, Yutao; An, Seungdo; Gianchandani, Yogesh B.

    2015-11-01

    This paper reports an evaluation and a comparison of two architectures for implementing Si-micromachined high-flow Knudsen pumps. Knudsen pumps, which operate on the principle of thermal transpiration, have been shown to have great promise for micro-scale gas phase fluidic systems such as micro gas chromatographs. Simultaneously achieving both a high flow rate and adequate blocking pressure has been a persistent challenge, which is addressed in this work by combining multiple pumps in series and addressing the resulting challenges in thermal management. The basic building block is a Si-micromachined pump with  ≈100 000 parallel channels in a 4 mm  ×  6 mm footprint. In the primary approach, multiple pump stages are stacked vertically with interleaved Si-micromachined spacers. A stacked 4-stage Knudsen pump has a form factor of 10 mm  ×  8 mm  ×  6 mm. In an alternate approach, multiple stages are arranged in a planar array. The experimental results demonstrate multiplication of the output pressure head with the number of stages, while the flow rate is maintained. For example, a stacked 4-stage Knudsen pump with 8 W power operated at atmospheric pressure provided a blocking pressure of 0.255 kPa, which was 3.6  ×  of that provided by a single-stage pump with 2 W power; while both provided a  ≈  30 sccm maximum flow rate. The performance can be customized for practical applications such as micro gas chromatography.

  14. Prediction of Shale Transport Properties Using the Lattice Boltzmann Method: Permeability and Effective Knudsen Diffusivity

    NASA Astrophysics Data System (ADS)

    Kang, Q.; Chen, L.

    2014-12-01

    Although short-term production of unconventional gas depends on the area of contact created by hydraulic fracturing and connections with pre-existing natural fracture networks, sustainable recovery is limited by transfer of gas from nanoporous matrix into the fractures, because the permeability of hydraulic fractures is orders of magnitude higher than that of the shale matrix. Therefore, a fundamental understanding of hydrocarbon mobility in shale matrix is urgently needed for improving recovery efficiencies. Shale transport properties (diffusivity, permeability, and electronic conductivity), which are critical for understanding the fundamental transport mechanisms, are still poorly understood. There have been some studies using experimental techniques such as scanning electron microscopy (SEM) to visualize the nanoscale structures of shale. Due to the ultra-low porosity and permeability, it is difficult to experimentally investigate the fundamental transport processes inside the shale or accurately measure the transport properties. Advanced pore-scale numerical methods, e.g., the lattice Boltzman method (LBM) may provide an alternative approach. In the present study, three-dimensional nanoscale porous structures of shale are reconstructed based on SEM images of shale samples. Characterization analysis of the nanoscale reconstructed shale is performed, including determination of porosity, pore size distribution, specific surface area, and pore connectivity. The LBM flow model and diffusion model are adopted to simulate fluid flow and Knudsen diffusion in the reconstructed shale, respectively. Tortuosity, intrinsic permeability, and effective Knudsen diffusivity are numerically predicted. The tortuosity is much higher than what is commonly employed in Bruggeman equation. Correction of the intrinsic permeability by taking into consideration the contribution of Knudsen diffusion, which leads to the apparent permeability, is performed. The correction factor under

  15. Use of finite volume radiation for predicting the Knudsen minimum in 2D channel flow

    SciTech Connect

    Malhotra, Chetan P.; Mahajan, Roop L.

    2014-12-09

    In an earlier paper we employed an analogy between surface-to-surface radiation and free-molecular flow to model Knudsen flow through tubes and onto planes. In the current paper we extend the analogy between thermal radiation and molecular flow to model the flow of a gas in a 2D channel across all regimes of rarefaction. To accomplish this, we break down the problem of gaseous flow into three sub-problems (self-diffusion, mass-motion and generation of pressure gradient) and use the finite volume method for modeling radiation through participating media to model the transport in each sub-problem as a radiation problem. We first model molecular self-diffusion in the stationary gas by modeling the transport of the molecular number density through the gas starting from the analytical asymptote for free-molecular flow to the kinetic theory limit of gaseous self-diffusion. We then model the transport of momentum through the gas at unit pressure gradient to predict Poiseuille flow and slip flow in the 2D gas. Lastly, we predict the generation of pressure gradient within the gas due to molecular collisions by modeling the transport of the forces generated due to collisions per unit volume of gas. We then proceed to combine the three radiation problems to predict flow of the gas over the entire Knudsen number regime from free-molecular to transition to continuum flow and successfully capture the Knudsen minimum at Kn ∼ 1.

  16. Overcoming the fill factor limit of double sided buried contact silicon solar cells

    NASA Astrophysics Data System (ADS)

    Ebong, A. U.; Lee, S. H.

    1997-11-01

    The double sided buried contact (DSBC) silicon solar cells have consistently demonstrated output parameters superior to those of its single sided counterparts. This is because the high surface recombination velocity of the single sided cells was reduced to minimum by the rear floating junction in conjunction with high quality silicon dioxide. However, the somewhat lower fill factor ( FF) exhibited by single sided illumination of the structure can disappear if the bifacial testing equipment is used to properly characterize the cells. A 2D simulation has shown that a rear illumination of only 0.2 sun in conjunction with 1 sun front illumination, is quite adequate to bias the rear junction to a high voltage required to maintain good fill factor. This article discusses the fill factor of double sided buried contact silicon solar cells under single and double sided illumination.

  17. Identification of a unique double-negative regulatory T-cell population.

    PubMed

    Lee, Byung O; Jones, Joyce E; Peters, Cory J; Whitacre, David; Frelin, Lars; Hughes, Janice; Kim, Won-Keun; Milich, David R

    2011-12-01

    Regulatory T (Treg) cells represent one of the main mechanisms of regulating self-reactive immune cells. Treg cells are thought to play a role in down-regulating immune responses to self or allogeneic antigens in the periphery. Although the function of Treg cells has been demonstrated in many experimental settings, the precise mechanisms and antigen specificity often remain unclear. In a hepatitis B e antigen-T-cell receptor (HBeAg-TCR) double transgenic mouse model, we observed a phenotypically unique (TCR+)  CD4- /CD8-  CD25(+/-)  GITR(high)  PD-1(high)  FoxP3-) HBeAg-specific population that demonstrates immune regulatory function. This HBeAg-specific double-negative regulatory cell population proliferates vigorously in vitro, in contrast to any other known regulatory population, in an interleukin-2-independent manner. PMID:22044159

  18. Identification of a unique double-negative regulatory T-cell population

    PubMed Central

    Lee, Byung O; Jones, Joyce E; Peters, Cory J; Whitacre, David; Frelin, Lars; Hughes, Janice; Kim, Won-Keun; Milich, David R

    2011-01-01

    Regulatory T (Treg) cells represent one of the main mechanisms of regulating self-reactive immune cells. Treg cells are thought to play a role in down-regulating immune responses to self or allogeneic antigens in the periphery. Although the function of Treg cells has been demonstrated in many experimental settings, the precise mechanisms and antigen specificity often remain unclear. In a hepatitis B e antigen–T-cell receptor (HBeAg-TCR) double transgenic mouse model, we observed a phenotypically unique (TCR+ CD4−/CD8− CD25+/− GITRhigh PD-1high FoxP3−) HBeAg-specific population that demonstrates immune regulatory function. This HBeAg-specific double-negative regulatory cell population proliferates vigorously in vitro, in contrast to any other known regulatory population, in an interleukin-2-independent manner. PMID:22044159

  19. High-rate lithium/manganese dioxide batteries; the double cell concept

    NASA Astrophysics Data System (ADS)

    Drews, Jürgen; Wolf, Rüdiger; Fehrmann, Gerd; Staub, Roland

    An implantable defibrillator battery has to provide pulse-power capabilities as well as high energy density. Low self-discharge rates are mandatory and an ability to check the state of charge is required. To accomplish these requirements, a lithium/manganese dioxide battery with a modified active cathode mass has been developed. Usage of a double cell design increases significantly the battery performance within an implantable defibrillator. The design features of a high-rate, pulse-power, manganese dioxide double cell are described.

  20. Simulation and optimization of current matching double-junction InGaN/Si solar cells

    NASA Astrophysics Data System (ADS)

    Nacer, S.; Aissat, A.

    2016-02-01

    This paper deals with theoretical investigation of the performance of current-matched In x GaN/Si double-junction solar cells. Calculations were performed under 1-sun AM1.5 using the one diode ideal model. Impact of minor carrier lifetime and surface recombination velocity in the top sub-cell on the cell performances is analyzed. Optimum composition of the top sub-cell has been identified ( x = 51.8 % and E g = 1.68 eV). The simulation results predict, for the optimized InGaN/Si double-junction solar cell, a short-circuit current J sc = 20 mA/cm2, an open-circuit voltage V oc = 1.97 V, and a conversion efficiency η = 38.3%.

  1. Multiple pathways of DNA double-strand break processing in a mutant Indian muntjac cell line

    SciTech Connect

    Bouffler, S.D.; Jha, B.; Johnson, R.T. )

    1990-09-01

    DNA break processing is compared in the Indian muntjac cell lines, SVM and DM. The initial frequencies and resealing of X-ray generated single- and double-strand breaks are similar in the two cell lines. Inhibiting the repair of UV damage leads to greater double-strand breakage in SVM than in DM, and some of these breaks are not repaired; however, repair-associated single-strand breakage and resealing are normal. Dimethylsulfate also induces excess double-strand breakage in SVM, and these breaks are irreparable. Restricted plasmids are reconstituted correctly in SVM at approximately 30% of the frequency observed in DM. Thus SVM has a reduced capacity to repair certain types of double-strand break. This defect is not due to a DNA ligase deficiency. We conclude that DNA double-strand breaks are repaired by a variety of pathways within mammalian cells and that the structure of the break or its mode of formation determines its subsequent fate.

  2. Investigating the validity of the Knudsen prescription for diffusivities in a mesoporous covalent organic framework

    SciTech Connect

    Krishna, Rajamani; van Baten, Jasper M.

    2011-04-27

    Molecular dynamics (MD) simulations were performed to determine the self-diffusivity (Di,self) and the Maxwell–Stefan diffusivity (ÐI) of hydrogen, argon, carbon dioxide, methane, ethane, propane, n-butane, n-pentane, and n-hexane in BTP-COF, which is a covalent organic framework (COF) that has one-dimensional 3.4-nm-sized channels. The MD simulations show that the zero-loading diffusivity (ÐI(0)) is consistently lower, by up to a factor of 10, than the Knudsen diffusivity (Di,Kn) values. The ratio ÐI(0)/Di,Kn is found to correlate with the isosteric heat of adsorption, which, in turn, is a reflection of the binding energy for adsorption on the pore walls: the stronger the binding energy, the lower the ratio ÐI(0)/Di,Kn. The diffusion selectivity, which is defined by the ratio D1,self/D2,self for binary mixtures, was determined to be significantly different from the Knudsen selectivity (M2/M1)1/2, where MI is the molar mass of species i. For mixtures in which component 2 is more strongly adsorbed than component 1, the expression (D1,self/D2,self)/(M2/M1)1/2 has values in the range of 1–10; the departures from the Knudsen selectivity increased with increasing differences in adsorption strengths of the constituent species. The results of this study have implications in the modeling of diffusion within mesoporous structures, such as MCM-41 and SBA-15.

  3. Double labeling autoradiography. Cell kinetic studies with /sup 3/H- and /sup 14/C-thymidine

    SciTech Connect

    Schultze, B.

    1981-01-01

    Examples of the multiple applicability of the double labeling method with /sup 3/H- and /sup 14/C-TdR are demonstrated. Double labeling with /sup 3/H- and /sup 14/C-TdR makes it possible to determine the cycle and its phases with high precision by modifying the usual percent labeled mitoses method with a single injection of /sup 3/H-TdR. In addition, data is provided on the variances of the transit times through the cycle phases. For example, in the case of the jejunal crypt cells of the mouse, the transit times through successive cycle phases are uncorrelated. In the case of glial cells the double labeling method provides cell kinetic parameters despite the paucity of proliferating glial cells. In the adult untreated animal, glial cell mitoses are so rare that the percent labeled mitoses method can not be utilized. However, the S-phase duration can be measured by double labeling and the cycle time can be determined by the so-called method of labeled S phases. With the latter method the passage through the S phase of the /sup 3/H-TdR-labeled S phase cells can be registered by injecting /sup 14/C-TdR at different time intervals following /sup 3/H-TdR application. In this way an S-phase duration of about 10 hr and a cycle time of about 20 hr was found for glial cells in the adult untreated mouse. An exchange of glial cells between the growth fraction and the nongrowth fraction has also been shown by double labeling. A quite different application of the double labeling method with 3H- and /sup 14/C-TdR is the in vivo study of the cell cycle phase-specific effect of drugs used in chemotherapy of tumors. The effect of vincristine on these cells has been studied. Vincristine affects cells in S and G2 in such a manner that they are arrested during the next metaphase and subsequently become necrotic. It has no effect on G1 cells.

  4. Vaporization study of sodium sulphate — potassium sulphate binary system by Knudsen effusion mass spectrometry

    NASA Astrophysics Data System (ADS)

    Armatys, K.; Miller, M.; Matraszek, A.; Wolter, A.

    2011-06-01

    The vaporization of samples of different chemical and phase compositions in the Na2SO4-K2SO4 system was investigated over the temperature range 1100 K-1400 K by the use of Knudsen effusion mass spectrometry. The gaseous species Na(g), Na2SO4(g), K(g), K2SO4(g), SO2(g), O2(g) and NaKSO4(g) were identified in the vapour over the samples investigated. The thermodynamic activities of sulphates in the examined system at 1350 K were obtained, which allowed calculating the chemical composition of the vapours present in the high temperature zone of cement kilns.

  5. Enhanced light trapping with double-groove grating in thin-film amorphous silicon solar cells

    NASA Astrophysics Data System (ADS)

    Wu, Jun

    2016-05-01

    A design to enhance light absorption in thin-film amorphous silicon (a-Si) solar cells is proposed. It is achieved by patterning a double-groove grating with waveguide layer as the absorbing layer and coating a double-groove grating anti-reflective layer in the front window of the cell. The broadband absorption under normal incidence can be achieved for both TE and TM polarizations. It is shown that the averaged integrated absorptions have very large angle independence for the optimized solar cell. An qualitative understanding of such broadband enhanced absorption effect, which is attributed to the guided mode resonance, is presented. The conclusions can be exploited to guide the design of solar cells based on a grating structure.

  6. Porous copper zinc tin sulfide thin film as photocathode for double junction photoelectrochemical solar cells.

    PubMed

    Dai, Pengcheng; Zhang, Guan; Chen, Yuncheng; Jiang, Hechun; Feng, Zhenyu; Lin, Zhaojun; Zhan, Jinhua

    2012-03-21

    Porous copper zinc tin sulfide (CZTS) thin film was prepared via a solvothermal approach. Compared with conventional dye-sensitized solar cells (DSSCs), double junction photoelectrochemical cells using dye-sensitized n-type TiO(2) (DS-TiO(2)) as the photoanode and porous p-type CZTS film as the photocathode shows an increased short circuit current, external quantum efficiency and power conversion efficiency. PMID:22322239

  7. One-dimensional particle simulations of Knudsen-layer effects on D-T fusion

    SciTech Connect

    Cohen, Bruce I.; Dimits, Andris M.; Zimmerman, George B.; Wilks, Scott C.

    2014-12-15

    Particle simulations are used to solve the fully nonlinear, collisional kinetic equation describing the interaction of a high-temperature, high-density, deuterium-tritium plasma with absorbing boundaries, a plasma source, and the influence of kinetic effects on fusion reaction rates. Both hydrodynamic and kinetic effects influence the end losses, and the simulations show departures of the ion velocity distributions from Maxwellian due to the reduction of the population of the highest energy ions (Knudsen-layer effects). The particle simulations show that the interplay between sources, plasma dynamics, and end losses results in temperature anisotropy, plasma cooling, and concomitant reductions in the fusion reaction rates. However, for the model problems and parameters considered, particle simulations show that Knudsen-layer modifications do not significantly affect the velocity distribution function for velocities most important in determining the fusion reaction rates, i.e., the thermal fusion reaction rates using the local densities and bulk temperatures give good estimates of the kinetic fusion reaction rates.

  8. Knudsen effusion mass spectrometric determination of mixing thermodynamic data of liquid Ag-In-Sn alloy

    NASA Astrophysics Data System (ADS)

    Bencze, L.; Popovic, A.

    2008-03-01

    The vaporisation of a liquid Ag-In-Sn system has been investigated at 1273-1473 K by Knudsen effusion mass spectrometry (KEMS) and the data fitted to a Redlich-Kister-Muggianu (RKM) sub-regular solution model. Nineteen different compositions have been examined at six fixed indium mole fractions, XIn = 0.10, 0.117, 0.20, 0.30, 0.40 and 0.50. The ternary L-parameters, the thermodynamic activities and the thermodynamic properties of mixing have been evaluated using standard KEMS procedures and from the measured ion intensity ratios of Ag+ to In+ and Ag+ to Sn+, using a mathematical regression technique described by us for the first time. The intermediate data obtained directly from the regression technique are the RKM ternary L-parameters. From the obtained ternary L-parameters the integral molar excess Gibbs free energy, the excess chemical potentials, the activity coefficients and the activities have been evaluated. Using the temperature dependence of the activities, the integral and partial molar excess enthalpies and entropies were determined. In addition, for comparison, for some compositions, also the Knudsen effusion isothermal evaporation method (IEM) and the Gibbs-Duhem ion intensity ratio method (GD-IIR) were used to determine activities and good agreement was obtained with the data obtained from fitting to the RKM model.

  9. Knudsen temperature jump and the Navier-Stokes hydrodynamics of granular gases driven by thermal walls.

    PubMed

    Khain, Evgeniy; Meerson, Baruch; Sasorov, Pavel V

    2008-10-01

    Thermal wall is a convenient idealization of a rapidly vibrating plate used for vibrofluidization of granular materials. The objective of this work is to incorporate the Knudsen temperature jump at thermal wall in the Navier-Stokes hydrodynamic modeling of dilute granular gases of monodisperse particles that collide nearly elastically. The Knudsen temperature jump manifests itself as an additional term, proportional to the temperature gradient, in the boundary condition for the temperature. Up to a numerical prefactor O(1) , this term is known from kinetic theory of elastic gases. We determine the previously unknown numerical prefactor by measuring, in a series of molecular dynamics (MD) simulations, steady-state temperature profiles of a gas of elastically colliding hard disks, confined between two thermal walls kept at different temperatures, and comparing the results with the predictions of a hydrodynamic calculation employing the modified boundary condition. The modified boundary condition is then applied, without any adjustable parameters, to a hydrodynamic calculation of the temperature profile of a gas of inelastic hard disks driven by a thermal wall. We find the hydrodynamic prediction to be in very good agreement with MD simulations of the same system. The results of this work pave the way to a more accurate hydrodynamic modeling of driven granular gases. PMID:18999414

  10. Zinc chromate induces chromosome instability and DNA double strand breaks in human lung cells

    SciTech Connect

    Xie Hong; Holmes, Amie L.; Young, Jamie L.; Qin Qin; Joyce, Kellie; Pelsue, Stephen C.; Peng Cheng; Wise, Sandra S.; Jeevarajan, Antony S.; Wallace, William T.; Hammond, Dianne; Wise, John Pierce E-mail: John.Wise@usm.maine.edu

    2009-02-01

    Hexavalent chromium Cr(VI) is a respiratory toxicant and carcinogen, with solubility playing an important role in its carcinogenic potential. Zinc chromate, a water insoluble or 'particulate' Cr(VI) compound, has been shown to be carcinogenic in epidemiology studies and to induce tumors in experimental animals, but its genotoxicity is poorly understood. Our study shows that zinc chromate induced concentration-dependent increases in cytotoxicity, chromosome damage and DNA double strand breaks in human lung cells. In response to zinc chromate-induced breaks, MRE11 expression was increased and ATM and ATR were phosphorylated, indicating that the DNA double strand break repair system was initiated in the cells. In addition, our data show that zinc chromate-induced double strand breaks were only observed in the G2/M phase population, with no significant amount of double strand breaks observed in G1 and S phase cells. These data will aid in understanding the mechanisms of zinc chromate toxicity and carcinogenesis.

  11. Tandem-structured, hot electron based photovoltaic cell with double Schottky barriers

    PubMed Central

    Lee, Young Keun; Lee, Hyosun; Park, Jeong Young

    2014-01-01

    We demonstrate a tandem-structured, hot electron based photovoltaic cell with double Schottky barriers. The tandem-structured, hot electron based photovoltaic cell is composed of two metal/semiconductor interfaces. Two types of tandem cells were fabricated using TiO2/Au/Si and TiO2/Au/TiO2, and photocurrent enhancement was detected. The double Schottky barriers lead to an additional pathway for harvesting hot electrons, which is enhanced through multiple reflections between the two barriers with different energy ranges. In addition, light absorption is improved by the band-to-band excitation of both semiconductors with different band gaps. Short-circuit current and energy conversion efficiency of the tandem-structured TiO2/Au/Si increased by 86% and 70%, respectively, compared with Au/Si metal/semiconductor nanodiodes, showing an overall solar energy conversion efficiency of 5.3%. PMID:24694838

  12. Double-Cell Geometry for 129Xe/3He Co-Magnetometry

    NASA Astrophysics Data System (ADS)

    Ohtomo, Yuichi; Ichikawa, Yuichi; Sato, Tomoya; Sakamoto, Yu; Kojima, Shuichiro; Suzuki, Takahiro; Shirai, Hazuki; Chikamori, Masatoshi; Hikota, Eri; Miyatake, Hirokazu; Nanao, Tsubasa; Suzuki, Kunifumi; Tsuchiya, Masato; Inoue, Takeshi; Furukawa, Takeshi; Yoshimi, Akihiro; Bidinosti, Christopher P.; Ino, Takashi; Ueno, Hideki; Matsuo, Yukari; Fukuyama, Takeshi; Asahi, Koichiro

    Comagnetometers play a key role in EDM experiments. They allow one to quantify, and subsequently correct for, any long-term drifts of the external magnetic field. In order to improve the performance of the 3He comagnetometer for our 129Xe EDM measurements, we have decided to incorporate a double-cell geometry which enables us to suppress a frequency shift due to contact interaction with polarized Rb atoms. In this study, the production and relaxation of 3He spin polarization in the double cell were studied. As a result, the followings were achieved: a polarization of 1.04(8)%, a longitudinal spin relaxation time of 10.1(5) h, and a transverse relaxation time of 2,340 s. With these improvements, concurrent operation of the 129Xe and 3He masers has been realized, and EDM measurement will be started in near future using a cell designed based on the results of this study.

  13. Breast Cancer Cell Lines Exhibit Differential Sensitivities to Microtubule-targeting Drugs Independent of Doubling Time

    PubMed Central

    RISINGER, APRIL L.; DYBDAL-HARGREAVES, NICHOLAS F.; MOOBERRY, SUSAN L.

    2016-01-01

    Background Microtubule-targeting agents (MTAs) are a mainstay in breast cancer treatment, yet patient responses differ. The underlying mechanisms of these differences are unknown. While MTAs are mitotic inhibitors, recent evidence highlights that non-mitotic effects of these drugs can contribute to their anticancer effects. It is critical to identify the non-mitotic mechanisms that could contribute to differences among MTAs. However, it is not clear whether rapidly dividing cells in culture are optimal tools to address these mechanistic questions in interphase cells. Materials and Methods Detailed concentration response curves for five MTAs in a panel of diverse breast cancer cell lines were generated. Results Substantial differences among both drugs and cell lines, consistent with the clinical scenario, were observed. Importantly, these differences do not correlate with cell doubling time. Conclusion The interphase actions of MTAs are critical to the full spectrum of their effects in cancer cells, even in cell culture models. PMID:26504006

  14. Engineering artificial cells by combining HeLa-based cell-free expression and ultra-thin double emulsion template

    PubMed Central

    Ho, Kwun Yin; Murray, Victoria L.; Liu, Allen P.

    2015-01-01

    Generation of artificial cells provides the bridge needed to cover the gap between studying the complexity of biological processes in whole cells and studying these same processes in an in vitro reconstituted system. Artificial cells are defined as the encapsulation of biologically active material in a biological or synthetic membrane. Here, we describe a robust and general method to produce artificial cells for the purpose of mimicking one or more behaviors of a cell. A microfluidic double emulsion system is used to encapsulate a mammalian cell free expression system that is able to express membrane proteins into the bilayer or soluble proteins inside the vesicles. The development of a robust platform that allows the assembly of artificial cells is valuable in understanding subcellular functions and emergent behaviors in a more cell-like environment as well as for creating novel signaling pathways to achieve specific cellular behaviors. PMID:25997354

  15. Increased CD4+/CD8+ Double-Positive T Cells in Chronic Chagasic Patients

    PubMed Central

    Giraldo, Nicolas A.; Bolaños, Natalia I.; Cuellar, Adriana; Guzman, Fanny; Uribe, Ana Maria; Bedoya, Astrid; Olaya, Natalia; Cucunubá, Zulma M.; Roa, Nubia; Rosas, Fernando; Velasco, Víctor; Puerta, Concepción J.; González, John M.

    2011-01-01

    Background CD4+/CD8+ double positive (DP) T cells have been described in healthy individuals as well as in patients with autoimmune and chronic infectious diseases. In chronic viral infections, this cell subset has effector memory phenotype and displays antigen specificity. No previous studies of double positive T cells in parasite infections have been carried out. Methodology/Principal Findings Seventeen chronic chagasic patients (7 asymptomatic and 10 symptomatic) and 24 non-infected donors, including 12 healthy and 12 with non-chagasic cardiomyopathy donors were analyzed. Peripheral blood was stained for CD3, CD4, CD8, HLA-DR and CD38, and lymphocytes for intracellular perforin. Antigen specificity was assessed using HLA*A2 tetramers loaded with T. cruzi K1 or influenza virus epitopes. Surface expression of CD107 and intracellular IFN-γ production were determined in K1-specific DP T cells from 11 chagasic donors. Heart tissue from a chronic chagasic patient was stained for both CD8 and CD4 by immunochemistry. Chagasic patients showed higher frequencies of DP T cells (2.1%±0.9) compared with healthy (1.1%±0.5) and non-chagasic cardiomyopathy (1.2%±0.4) donors. DP T cells from Chagasic patients also expressed more HLA-DR, CD38 and perforin and had higher frequencies of T. cruzi K1-specific cells. IFN-γ production in K1-specific cells was higher in asymptomatic patients after polyclonal stimulation, while these cells tended to degranulate more in symptomatic donors. Immunochemistry revealed that double positive T cells infiltrate the cardiac tissue of a chagasic donor. Conclusions Chagasic patients have higher percentages of circulating double positive T cells expressing activation markers, potential effector molecules and greater class I antigenic specificity against T. cruzi. Although K1 tetramer positive DP T cell produced little IFN-γ, they displayed degranulation activity that was increased in symptomatic patients. Moreover, K1-specific DP T cells can

  16. Canine CD4(+)CD8(+) double-positive T cells can develop from CD4(+) and CD8(+) T cells.

    PubMed

    Bismarck, Doris; Moore, Peter F; Alber, Gottfried; von Buttlar, Heiner

    2014-12-15

    For a long time the expression of the CD4 and CD8 receptor on peripheral blood T cells was thought to be mutually exclusive. However, in canine peripheral blood, similar to other species as swine or human for example, mature CD4(+)CD8(+) double-positive (dp) T cells exist which simultaneously express both surface receptors and have features of activated T cells. Canine CD4(+)CD8(+)dp T cells are heterogeneous and can be divided into three subpopulations by their intensity of CD4 and CD8α expression: CD4(bright)CD8α(bright), CD4(dim)CD8α(bright) and CD4(dim)CD8α(dim). The number of CD4(+)CD8α(+)dp T cells increases after in vitro stimulation of canine peripheral blood mononuclear cells (PBMC) raising the question of their progenitor(s). Thus, the aim of our study was to characterize the progenitor(s) of canine CD4(+)CD8α(+)dp T cells. By cell tracing experiments we identified both CD4(+) single-positive (sp) and also CD8α(+)sp T cells as progenitors of canine CD4(+)CD8α(+)dp T cells after in vitro stimulation. CD4(+)sp T cells almost exclusively upregulate a CD8αα homodimer, whereas CD8α(+)sp T cells can become CD4(+)CD8αβ(+) or CD4(+)CD8αα(+). Even in the absence of other cells, highly purified CD4(+)sp T cells can become double-positive upon in vitro stimulation, whereas highly purified CD8α(+)sp T cells fail to do so. However, CD8α(+)sp T cells can additionally express CD4 when stimulated in the presence of CD4(-)CD8α(-) double-negative (dn) cells or more efficiently when stimulated in the presence of CD4(+)sp T cells. Soluble factors secreted by CD4(+)sp T cells are sufficient for the upregulation of CD4 on CD8α(+)sp T cells, but direct cell-cell contact between CD4(+)sp and CD8α(+)sp T cells is more efficient. mRNA analysis shows that additional CD4 expression on CD8α(+)sp T cells results from de novo synthesis. Thus, uptake of soluble CD4 or trogocytosis is less likely as mechanism for generation of canine double-positive T cells. CD4(+)CD

  17. GABAergic and glycinergic pathways to goldfish retinal ganglion cells: an ultrastructural double label study

    SciTech Connect

    Muller, J.F.

    1987-01-01

    An ultrastructural double label has been employed to compare GABAergic and glycinergic systems in the inner plexiform layer (IPL) of the goldfish retina. Electron microscope autoradiography of /sup 3/H-GABA and /sup 3/H-glycine uptake was combined with retrograde HRP-labeling of ganglion cells. When surveyed for distribution, GABAergic and glycinergic synapses were found onto labeled ganglion cells throughout the IPL. This reinforces previous physiological work that described GABAergic and glycinergic influences on a variety of ganglion cells in goldfish and carp; These physiological effects often reflect direct inputs.

  18. Efficiency enhancement of planar perovskite solar cells by adding zwitterion/LiF double interlayers for electron collection

    NASA Astrophysics Data System (ADS)

    Sun, Kuan; Chang, Jingjing; Isikgor, Furkan Halis; Li, Pengcheng; Ouyang, Jianyong

    2014-12-01

    Double interlayers consisting of a zwitterionic small molecule layer and a LiF layer were introduced between the electron transport layer and the cathode of perovskite solar cells. The double interlayers improve the photovoltaic efficiency to 13.2%, which is higher than that of control devices without the double interlayer (9.2%) or with LiF (11.0%) or rhodamine 101 zwitterion (12.1%) alone.Double interlayers consisting of a zwitterionic small molecule layer and a LiF layer were introduced between the electron transport layer and the cathode of perovskite solar cells. The double interlayers improve the photovoltaic efficiency to 13.2%, which is higher than that of control devices without the double interlayer (9.2%) or with LiF (11.0%) or rhodamine 101 zwitterion (12.1%) alone. Electronic supplementary information (ESI) available. See DOI: 10.1039/c4nr05975a

  19. Double-layered ZnO nanostructures for efficient perovskite solar cells

    NASA Astrophysics Data System (ADS)

    Mahmood, Khalid; S. Swain, Bhabani; Amassian, Aram

    2014-11-01

    To date, a single layer of TiO2 or ZnO has been the most successful implementations of any electron transport layer (ETL) in solution-processed perovskite solar cells. In a quest to improve the ETL, we explore a new nanostructured double-layer ZnO film for mesoscopic perovskite-based thin film photovoltaics. This approach yields a maximum power conversion efficiency of 10.35%, which we attribute to the morphology of oxide layer and to faster electron transport. The successful implementation of the low-temperature hydrothermally processed double-layer ZnO film as ETL in perovskite solar cells highlights the opportunities to further improve the efficiencies by focusing on the ETL in this rapidly developing field.

  20. Dye-sensitized solar cells using double-oxide electrodes: a brief review

    NASA Astrophysics Data System (ADS)

    Suzuki, Yoshikazu; Okamoto, Yuji; Ishii, Natsumi

    2015-04-01

    Dye-sensitized solar cells (DSC or DSSC) have been widely investigated because of their potentially high cost performance compared with Si-based solar cells and of their fascinating appearance. DSC with photoelectric conversion efficiency of >10 % (or even 12 %) have been reported, where porous TiO2 films are generally used as semi-conductor electrodes. Such porous TiO2 films usually have high specific surface area, and thus, they adsorb plenty of dye molecules, resulting in high photocurrent density. Recently, some double oxides have been examined as alternative photoanode materials, mainly in order to improve photovoltage. Here, studies on DSC using double-oxide electrodes, i.e., perovskite, spinel, ilmenite, wolframite, scheelite and pseudobrookite-types, are briefly reviewed.

  1. Double-layered ZnO nanostructures for efficient perovskite solar cells.

    PubMed

    Mahmood, Khalid; S Swain, Bhabani; Amassian, Aram

    2014-12-21

    To date, a single layer of TiO2 or ZnO has been the most successful implementations of any electron transport layer (ETL) in solution-processed perovskite solar cells. In a quest to improve the ETL, we explore a new nanostructured double-layer ZnO film for mesoscopic perovskite-based thin film photovoltaics. This approach yields a maximum power conversion efficiency of 10.35%, which we attribute to the morphology of oxide layer and to faster electron transport. The successful implementation of the low-temperature hydrothermally processed double-layer ZnO film as ETL in perovskite solar cells highlights the opportunities to further improve the efficiencies by focusing on the ETL in this rapidly developing field. PMID:25373624

  2. Exact calculation of the tortuosity in disordered linear pores in the Knudsen regime.

    PubMed

    Russ, Stefanie

    2009-12-01

    The squared reciprocal tortuosity kappa-2=D/D0 for linear diffusion on lattices and in pores in the Knudsen regime is calculated analytically for a large variety of disordered systems. Here, D0 and D are the self-diffusion coefficients of the smooth and the corresponding disordered system, respectively. To this end, a building-block principle is developed that composes the systems into substructures without cross correlations between them. It is shown how the solutions of the different building blocks can be combined to gain D/D0 for pores of high complexity from the geometrical properties of the systems, i.e., from the volumes of the different substructures. As a test, numerical simulations are performed that agree perfectly with the theory. PMID:20365144

  3. Slip-flow experiments in welded tuff: the Knudsen diffusion problem

    SciTech Connect

    Reda, D.C.

    1985-12-31

    Gas-phase permeability experiments, using nitrogen, were conducted on a sample of densely welded tuffaceous material from the Nevada Test Site. The primary independent variable was the average pore pressure, which was systematically varied from 13.1 to 0.1 MPa; net circumferential stress was maintained constant at 4.3 MPa. Gas permeability was found to be a strong function of pore pressure, undergoing an order-of-magnitude increase as pore pressure was reduced over the stated range. Based on these measurements, the Knudsen diffusion coefficient for water vapor, at 296 K, was calculated to be about 3.2 x 10{sup -7} m{sup 2}/s and the material`s average pore diameter was estimated to be about 8 x 10{sup -9} m. Comparison of this length scale with mercury intrusion porosimetry results showed agreement to within one order of magnitude. 8 refs., 3 figs.

  4. Slip-flow experiments in welded tuff; the Knudsen diffusion problem

    SciTech Connect

    Reda, D.C.

    1985-09-01

    Gas-phase permeability experiments, using nitrogen, were conducted on a sample of densely welded tuffaceous material from the Nevada Test Site. The primary independent variable was the average pore pressure, which was systematically varied from 13.1 to 0.1 MPa; net circumferential stress was maintained constant at 4.3 MPa. Gas permeability was found to be a strong function of pore pressure, undergoing an order-of-magnitude increase as pore pressure was reduced over the stated range. Based on these measurements, the Knudsen diffusion coefficient for water vapor, at 296 K, was calculated to be {approx} 3.2 x 10{sup -7} m{sup 2}/s and the material`s average pore diameter was estimated to be {approx} 8 x 10{sup -9} m. Comparison of this length scale with mercury intrusion porosimetry results showed agreement to within one order of magnitude.

  5. Slip-flow experiments in welded tuff; The Knudsen diffusion problem

    SciTech Connect

    Reda, D.C.

    1987-12-31

    Gas-phase permeability experiments, using nitrogen, were conducted on a sample of densely welded tuffaceous material from the Nevada Test Site. The primary independent variable was the average pore pressure, which was systematically varied from 13.1 to 0.1 MPa; net circumferential stress was maintained constant at 4.3 MPa. Gas permeability was found to be a strong function of pore pressure, undergoing an order-of-magnitude increase as pore pressure was reduced over the stated range. Based on these measurements, the Knudsen diffusion coefficient for water vapor, at 296 K, was calculated to be {approx}3.2 {times} 10{sup {minus}7} m{sup 2}/s and the material`s average pore diameter was estimated to be {approx}8 {times} 10{sup {minus}9} m. Comparison of this length scale with mercury intrusion porosimetry results shows agreement to within one order of magnitude.

  6. Drag coefficient of the weakly ionized plasma in the high Knudsen number regime

    SciTech Connect

    Chu, H.-Y.; Si, M.-C.; Lin, S.-B.

    2009-06-15

    The drag force acting on a micron-sized polystyrene particle in the high Knudsen number regime is investigated. Analysis of the particle trajectories in stationary neutral argon gas environment suggests the damping time constant {tau}{proportional_to}p{sup -1.20{+-}}{sup 0.04} and Epstein drag force coefficient {delta}=1.40. The neutral drag coefficient is compared with the drag coefficient measurement in dust-free plasma. The phenomena of the reduced drag in weakly viscous and weakly ionized rf plasma are also observed in this report. It is shown that the slight changes in rf power and pressure would enhance the reduced drag effect, which suggests that there is an additional electrostatic force acting along the particle motion in the plasma.

  7. A method for double-labeling sputum cells for p53 and cytokeratin

    SciTech Connect

    Neft, R.E.; Tierney, L.A.; Belinsky, S.A.

    1995-12-01

    Molecular and immunological techniques may enhance the usefulness of sputum cytology as a screening tool for lung cancer. These techniques may also be useful in detecting and following the early progression of disease from metaplasia to dysplasia, carcinoma in situ, and finally to invasive carcinoma. Longitudinal information on the evolution of these malignant changes in the respiratory epithelium can be gained by prospective study of populations at high risk for lung cancer. This work is significant because double-labeling of cells in sputum with p53 and cytokeratin antibodies facilitates rapid screening of p53 positive neoplastic and preneoplastic lung cells by brightfield and fluorescence microscopy.

  8. Double Negative (DN) αβ T Cells: misperception and overdue recognition

    PubMed Central

    Martina, Maria N; Noel, Sanjeev; Saxena, Ankit; Rabb, Hamid; Hamad, Abdel Rahim A

    2015-01-01

    CD4−CD8− double negative (DN) αβ T cells are legitimate components of the normal immune system. However, they are poorly understood and largely ignored by immunologists because of their historical association with the lymphoproliferation that occurs in mice (lpr and gld) and humans (ALPS patients) with impaired Fas-mediated apoptosis where they are considered abnormal T cells. We believe that the traditional view that DN T cells that cause lymphoproliferation (hereafter referred to as lpr DN T cells) are CD4 and CD8 T cells that lost their coreceptor, conceived more than two decades ago, is flawed and that conflating lpr DN T cells with DN T cells found in normal immune system (hereafter referred to as nDN T cells) is unnecessarily dampening interest of this potentially important cell type. To begin rectifying these misperceptions, we will revisit the traditional view of lpr DN T cells and show that it does not hold true in light of recent immunological advances. In lieu of it, we offer a new model proposing that Fas-mediated apoptosis actively removes normally existing DN T cells from the periphery and that impaired Fas-mediated apoptosis leads to accumulation of these cells rather than de novo generation of DN T cells from activated CD4 or CD8 T cells. By doing so, we hope to provoke a new discussion that may lead to a consensus about the origin of lpr DN T cells and regulation of their homeostasis by the Fas pathway and reignite wider interest in nDN T cells. PMID:25420721

  9. Canine CD4+CD8+ double positive T cells in peripheral blood have features of activated T cells.

    PubMed

    Bismarck, Doris; Schütze, Nicole; Moore, Peter; Büttner, Mathias; Alber, Gottfried; Buttlar, Heiner v

    2012-10-15

    In dogs a CD4(+)CD8(+) double positive T cell subpopulation exists that has not been phenotypically defined yet. We demonstrate that canine CD4(+)CD8(+) T cells are mature CD1a(-) and TCRαβ(+) T cells. To analyse the activation potential of CD4(+)CD8(+) T cells, PBMC from dogs vaccinated against canine distemper virus (CDV) were re-stimulated with CDV. Upon antigen-specific stimulation, the CD4(+)CD8(+) T cell fraction increases and consists nearly exclusively of proliferated cells. Similarly, other features of activated effector/memory T cells such as up-regulation of CD25 and MHC-II as well as down-regulation of CD62L (L-selectin) were observed in CD4(+)CD8(+) T cells after stimulation. Canine CD4(+)CD8(+) T cells are less abundant, but more heterogeneous than porcine ones, comprising a small proportion expressing the β chain of CD8 in addition to the CD8α chain, like human CD4(+)CD8(+) T cells. In summary, this analysis provides the basis for functional characterisation of the in vivo relevance of CD4(+)CD8(+) T cells in T-cell mediated immunity. PMID:22789871

  10. A microchip electrophoresis-mass spectrometric platform with double cell lysis nano-electrodes for automated single cell analysis.

    PubMed

    Li, Xiangtang; Zhao, Shulin; Hu, Hankun; Liu, Yi-Ming

    2016-06-17

    Capillary electrophoresis-based single cell analysis has become an essential approach in researches at the cellular level. However, automation of single cell analysis has been a challenge due to the difficulty to control the number of cells injected and the irreproducibility associated with cell aggregation. Herein we report the development of a new microfluidic platform deploying the double nano-electrode cell lysis technique for automated analysis of single cells with mass spectrometric detection. The proposed microfluidic chip features integration of a cell-sized high voltage zone for quick single cell lysis, a microfluidic channel for electrophoretic separation, and a nanoelectrospray emitter for ionization in MS detection. Built upon this platform, a microchip electrophoresis-mass spectrometric method (MCE-MS) has been developed for automated single cell analysis. In the method, cell introduction, cell lysis, and MCE-MS separation are computer controlled and integrated as a cycle into consecutive assays. Analysis of large numbers of individual PC-12 neuronal cells (both intact and exposed to 25mM KCl) was carried out to determine intracellular levels of dopamine (DA) and glutamic acid (Glu). It was found that DA content in PC-12 cells was higher than Glu content, and both varied from cell to cell. The ratio of intracellular DA to Glu was 4.20±0.8 (n=150). Interestingly, the ratio drastically decreased to 0.38±0.20 (n=150) after the cells are exposed to 25mM KCl for 8min, suggesting the cells released DA promptly and heavily while they released Glu at a much slower pace in response to KCl-induced depolarization. These results indicate that the proposed MCE-MS analytical platform may have a great potential in researches at the cellular level. PMID:27207575

  11. X-ray induced DNA double strand break production and repair in mammalian cells as measured by neutral filter elution.

    PubMed Central

    Bradley, M O; Kohn, K W

    1979-01-01

    This work presents a neutral filter elution method for detecting DNA double strand breaks in mouse L1210 cells after X-ray. The assay will detect the number of double strand breaks induced by as little as 1000 rad of X-ray. The rate of DNA elution through the filters under neutral conditions increases with X-ray dose. Certain conditions for deproteinization, pH, and filter type are shown to increase the assay's sensitivity. Hydrogen peroxide and Bleomycin also induce apparent DNA double strand breaks, although the ratios of double to single strand breaks vary from those produced by X-ray. The introduction of double strand cuts by HpA I restriction endonuclease in DNA lysed on filters results in a rapid rate of elution under neutral conditions, implying that the method can detect double strand breaks if they exist in the DNA. The eluted DNA bands with a double stranded DNA marker in cesium chloride. This evidence suggests that the assay detects DNA double strand breaks. L1210 cells are shown to rejoin most of the DNA double strand breaks induced by 5-10 krad of X-ray with a half-time of about 40 minutes. PMID:92010

  12. Inactivation, DNA double strand break induction and their rejoining in bacterial cells irradiated with heavy ions

    NASA Technical Reports Server (NTRS)

    Schaefer, M.; Zimmermann, H.; Schmitz, C.

    1994-01-01

    Besides inactivation one of the major interests in our experiments is to study the primary damage in the DNA double strand breaks (DSB) after heavy ion irradiation. These damages lead not only to cell death but also under repair activities to mutations. In further experiments we have investigated the inactivation with two different strains of Deinococcus radiodurans (R1, Rec 30) and the induction of DSB as well as the rejoining of DSB in stationary cells of E. coli (strain B/r) irradiated with radiations of different quality. In the latter case irradiations were done so that the cell survival was roughly at the same level. We measured the DSB using the pulse field gelelectrophoresis which allows to separate between intact (circular) and damaged (linear) DNA. The irradiated cells were transferred to NB medium and incubated for different times to allow rejoining.

  13. Functional expression of double-stranded RNA-dependent protein kinase in rat intestinal epithelial cells.

    PubMed

    Sato, Nagahiro; Morimoto, Hiroyuki; Baba, Ryoko; Nakamata, Junichi; Doi, Yoshiaki; Yamaguchi, Koji

    2010-05-01

    Intestinal epithelial cells (IECs) are exposed to external environment, microbial and viral products, and serve as essential barriers to antigens. Recent studies have shown that IECs express Toll-like receptors (TLRs) and respond to microbial components. The antimicrobial and antiviral barriers consist of many molecules including TLRs. To investigate the further component of this barrier in intestine, we examined the expression of double-stranded RNA-dependent protein kinase (PKR). PKR is a player in the cellular antiviral response and phosphorylates alpha-subunit of the eukaryotic translation initiation factor 2 (eIF-2alpha) to block protein synthesis and induces apoptosis. In this study, we showed that the expression of PKR was restricted to the cytoplasm of absorptive epithelial cells in the intestine of adult rat. We also demonstrated that PKR was expressed in the cultured rat intestinal epithelial cells (IEC-6). The level of PKR protein expression and the activity of alkaline phosphatase (ALP) increased in the cultured IEC-6 cells in a time-dependent manner. Inhibition of PKR by the 2-aminopurine treatment decreased ALP activity in the IEC-6 cells. Treatment of IEC-6 cells with synthetic double-stranded RNA (dsRNA) induced cell death in a dose-dependent manner. The addition of hydrocortisone also provoked suppression of PKR expression and ALP activity. This modulation might be mediated by signal transducers and activators of transcription-1 (STAT-1) protein. We concluded that PKR is expressed in IECs as potent barriers to antigens and is a possible modulator of the differentiation of rat IECs. PMID:20213745

  14. Double Strain-Promoted Macrocyclization for the Rapid Selection of Cell-Active Stapled Peptides.

    PubMed

    Lau, Yu Heng; Wu, Yuteng; Rossmann, Maxim; Tan, Ban Xiong; de Andrade, Peterson; Tan, Yaw Sing; Verma, Chandra; McKenzie, Grahame J; Venkitaraman, Ashok R; Hyvönen, Marko; Spring, David R

    2015-12-14

    Peptide stapling is a method for designing macrocyclic alpha-helical inhibitors of protein-protein interactions. However, obtaining a cell-active inhibitor can require significant optimization. We report a novel stapling technique based on a double strain-promoted azide-alkyne reaction, and exploit its biocompatibility to accelerate the discovery of cell-active stapled peptides. As a proof of concept, MDM2-binding peptides were stapled in parallel, directly in cell culture medium in 96-well plates, and simultaneously evaluated in a p53 reporter assay. This in situ stapling/screening process gave an optimal candidate that showed improved proteolytic stability and nanomolar binding to MDM2 in subsequent biophysical assays. α-Helicity was confirmed by a crystal structure of the MDM2-peptide complex. This work introduces in situ stapling as a versatile biocompatible technique with many other potential high-throughput biological applications. PMID:26768531

  15. Layered Double Hydroxide as a Vehicle to Increase Toxicity of Gallate Ions against Adenocarcinoma Cells.

    PubMed

    Arratia-Quijada, Jenny; Rivas-Fuentes, Selma; Saavedra, Karina J Parra; Lamas, Adriana M Macías; Carbajal Arízaga, Gregorio Guadalupe

    2016-01-01

    The antineoplasic activity of gallic acid has been reported. This compound induces apoptosis and inhibits the growth of several neoplasic cells. However, this molecule is easily oxidized and degraded in the body. The aim of this work was to intercalate gallate ions into layered double hydroxide (LDH) nanoparticles under controlled conditions to reduce oxidation of gallate and to evaluate its toxicity against the A549 adenocarcinoma cell line. An isopropanol medium under nitrogen atmosphere was adequate to intercalate gallate ions with a lesser oxidation degree as detected by electron spin resonance spectroscopy. Concentrations of the hybrid LDH-gallate nanoparticles between 0.39 and 25 µg/mL reduced the cell viability to 67%, while the value reached with the pure gallic acid and LDH was 90% and 78%, respectively, thus proving that the combination of gallate ions with the inorganic nanoparticles increases the toxicity potential within this dose range. PMID:27438820

  16. Broadband Absorption Enhancement in Thin Film Solar Cells Using Asymmetric Double-Sided Pyramid Gratings

    NASA Astrophysics Data System (ADS)

    Alshal, Mohamed A.; Allam, Nageh K.

    2016-07-01

    A design for a highly efficient modified grating crystalline silicon (c-Si) thin film solar cell is demonstrated and analyzed using the two-dimensional (2-D) finite element method. The suggested grating has a double-sided pyramidal structure. The incorporation of the modified grating in a c-Si thin film solar cell offers a promising route to harvest light into the few micrometers active layer. Furthermore, a layer of silicon nitride is used as an antireflection coating (ARC). Additionally, the light trapping through the suggested design is significantly enhanced by the asymmetry of the top and bottom pyramids. The effects of the thickness of the active layer and facet angle of the pyramid on the spectral absorption, ultimate efficiency (η), and short-circuit current density (J sc) are investigated. The numerical results showed 87.9% efficiency improvement over the conventional thin film c-Si solar cell counterpart without gratings.

  17. The Mean-field Solar Dynamo with a Double Cell Meridional Circulation Pattern

    NASA Astrophysics Data System (ADS)

    Pipin, V. V.; Kosovichev, A. G.

    2013-10-01

    Recent helioseismology findings, as well as advances in direct numerical simulations of global dynamics of the Sun, have indicated that in each solar hemisphere meridional circulation may form more than one cell along the radius in the convection zone. In particular, recent helioseismology results revealed a double-cell structure of the meridional circulation. We investigate properties of a mean-field solar dynamo with such double-cell meridional circulation. The dynamo model also includes the realistic profile of solar differential rotation (including the tachocline and subsurface shear layer) and takes into account effects of turbulent pumping, anisotropic turbulent diffusivity, and conservation of magnetic helicity. Contrary to previous flux-transport dynamo models, we find that the dynamo model can robustly reproduce the basic properties of the solar magnetic cycles for a wide range of model parameters and circulation speeds. The best agreement with observations is achieved when the surface meridional circulation speed is about 12 m s-1. For this circulation speed, the simulated sunspot activity shows good synchronization with the polar magnetic fields. Such synchronization was indeed observed during previous sunspot Cycles 21 and 22. We compare theoretical and observed phase diagrams of the sunspot number and the polar field strength and discuss the peculiar properties of Cycle 23.

  18. Numerical simulations of high Knudsen number gas flows and microchannel electrokinetic liquid flows

    NASA Astrophysics Data System (ADS)

    Yan, Fang

    Low pressure and microchannel gas flows are characterized by high Knudsen numbers. Liquid flows in microchannels are characterized by non-conventional driving potentials like electrokinetic forces. The main thrust of the dissertation is to investigate these two different kinds of flows in gases and liquids respectively. High Knudsen number (Kn) gas flows were characterized by 'rarified' or 'microscale' behavior. Because of significant non-continuum effect, traditional CFD techniques are often inaccurate for analyzing high Kn number gas flows. The direct simulation Monte Carlo (DSMC) method offers an alternative to traditional CFD which retains its validity in slip and transition flow regimes. To validate the DSMC code, comparisons of simulation results with theoretical analysis and experimental data are made. The DSMC method was first applied to compute low pressure, high Kn flow fields in partially heated two dimensional channels. The effects of varying pressure, inlet flow and gas transport properties (Kn, Reynolds number, Re and the Prandtl number, Pr respectively) on the wall heat transfer (Nusselt number, Nu) were examined. The DSMC method was employed to explore mixing gas flows in two dimensional microchannels. Mixing of two gas streams (H2 and O2) was considered within a microchannel. The effect of the inlet-outlet pressure difference, the pressure ratio of the incoming streams and the accommodation coefficient of the solid wall on mixing length were all examined. Parallelization of a three-dimensional DSMC code was implemented using OpenMP procedure on a shared memory multi-processor computer. The parallel code was used to simulate 3D high Kn number Couette flow and the flow characteristics are found to be very different from their continuum counterparts. A mathematical model describing electrokinetically driven mass transport phenomena in microfabricated chip devices will also be presented. The model accounts for the principal physical phenomena affecting

  19. Proliferation of Double-Strand Break-Resistant Polyploid Cells Requires Drosophila FANCD2.

    PubMed

    Bretscher, Heidi S; Fox, Donald T

    2016-06-01

    Conserved DNA-damage responses (DDRs) sense genome damage and prevent mitosis of broken chromosomes. How cells lacking DDRs cope with broken chromosomes during mitosis is poorly understood. DDRs are frequently inactivated in cells with extra genomes (polyploidy), suggesting that study of polyploidy can reveal how cells with impaired DDRs/genome damage continue dividing. Here, we show that continued division and normal organ development occurs in polyploid, DDR-impaired Drosophila papillar cells. As papillar cells become polyploid, they naturally accumulate broken acentric chromosomes but do not apoptose/arrest the cell cycle. To survive mitosis with acentric chromosomes, papillar cells require Fanconi anemia proteins FANCD2 and FANCI, as well as Blm helicase, but not canonical DDR signaling. FANCD2 acts independently of previous S phases to promote alignment and segregation of acentric DNA produced by double-strand breaks, thus avoiding micronuclei and organ malformation. Because polyploidy and impaired DDRs can promote cancer, our findings provide insight into disease-relevant DNA-damage tolerance mechanisms. PMID:27270041

  20. Molecular recombination and the repair of DNA double-strand breaks in CHO cells.

    PubMed Central

    Resnick, M A; Moore, P D

    1979-01-01

    Molecular recombination and the repair of DNA double-strand breaks (DSB) have been examined in the G-0 and S phase of the cell cycle using a temperature-sensitive CHO cell line to test i) if there are cell cycle restrictions on the repair of DSB's' ii) the extent to which molecular recombination can be induced between either sister chromatids or homologous chromosomes and iii) whether repair of DSB's involves recombination (3). Mitomycin C (1-2 micrograms/ml) or ionizing radiation (50 krad) followed by incubation resulted in molecular recombination (hybrid DNA) in S phase cells. Approximately 0.03 to 0.10% of the molecules (number average molecular weight: 5.6 x 10(6) Daltons after shearing) had hybrid regions for more than 75% of their length. However, no recombination was detected in G-0 cells. Since the repair of DSB was observed in both stages with more than 50% of the breaks repaired in 5 hours, it appears that DSB repair in G-0 cells does not involve recombination between homologous chromosomes. The possibility is not excluded that repair in G-0 cells involves only small regions (less than 4 x 10(6) Daltons). PMID:493136

  1. Simulation of the Formation of DNA Double Strand Breaks and Chromosome Aberrations in Irradiated Cells

    NASA Technical Reports Server (NTRS)

    Plante, Ianik; Ponomarev, Artem L.; Wu, Honglu; Blattnig, Steve; George, Kerry

    2014-01-01

    The formation of DNA double-strand breaks (DSBs) and chromosome aberrations is an important consequence of ionizing radiation. To simulate DNA double-strand breaks and the formation of chromosome aberrations, we have recently merged the codes RITRACKS (Relativistic Ion Tracks) and NASARTI (NASA Radiation Track Image). The program RITRACKS is a stochastic code developed to simulate detailed event-by-event radiation track structure: [1] This code is used to calculate the dose in voxels of 20 nm, in a volume containing simulated chromosomes, [2] The number of tracks in the volume is calculated for each simulation by sampling a Poisson distribution, with the distribution parameter obtained from the irradiation dose, ion type and energy. The program NASARTI generates the chromosomes present in a cell nucleus by random walks of 20 nm, corresponding to the size of the dose voxels, [3] The generated chromosomes are located within domains which may intertwine, and [4] Each segment of the random walks corresponds to approx. 2,000 DNA base pairs. NASARTI uses pre-calculated dose at each voxel to calculate the probability of DNA damage at each random walk segment. Using the location of double-strand breaks, possible rejoining between damaged segments is evaluated. This yields various types of chromosomes aberrations, including deletions, inversions, exchanges, etc. By performing the calculations using various types of radiations, it will be possible to obtain relative biological effectiveness (RBE) values for several types of chromosome aberrations.

  2. A double filtering method for measuring the translational velocity of fluorescently stained cells

    SciTech Connect

    Yasokawa, Toshiki; Ishimaru, Ichirou; Kuriyama, Shigeki; Masaki, Tsutomu; Takegawa, Kaoru; Tanaka, Naotaka

    2007-09-24

    The authors propose a double filtering method to measure translational velocity for tracking fluorescently stained cells. This method employs two diffraction gratings installed in the infinity space through which the parallel pencil beam of the fluorescence passes. With this method, the change in light intensity whose period is proportional to the translational velocity of the sample can be obtained at the imaging surface. By using a sample that has a random distribution of fluorescence intensity, the authors verified that translational velocity measurements could be achieved using the proposed method.

  3. General and Virus-Specific Immune Cell Reconstitution after Double Cord Blood Transplantation.

    PubMed

    Saliba, Rima M; Rezvani, Katayoun; Leen, Ann; Jorgensen, Jeffrey; Shah, Nina; Hosing, Chitra; Parmar, Simrit; Oran, Betul; Olson, Amanda; Rondon, Gabriela; Chen, Julianne; Martinez, Charles; Hamdi, Amir; Mehta, Rohtesh S; Chemaly, Roy F; Saunders, Ila M; Bollard, Catherine M; Shpall, Elizabeth J

    2015-07-01

    Cord blood transplantation (CBT) is curative for many patients with hematologic malignancies but is associated with delayed immune recovery and an increased risk of viral infections compared with HLA-matched bone marrow or peripheral blood progenitor cell transplantation. In this study we evaluated the significance of lymphocyte recovery in 125 consecutive patients with hematologic malignancies who underwent double-unit CBT (DUCBT) with an antithymocyte globulin-containing regimen at our institution. A subset of 65 patients was prospectively evaluated for recovery of T, natural killer (NK), and B cells, and in 46 patients we also examined viral-specific T cell recovery against adenovirus, Epstein-Barr virus, cytomegalovirus, BK virus, respiratory syncytial virus, and influenza antigen. Our results indicate that in recipients of DUCBT, the day 30 absolute lymphocyte count is highly predictive of nonrelapse mortality and overall survival. Immune recovery post-DUCBT was characterized by prolonged CD8+ and CD4+ T lymphopenia associated with preferential expansion of B and NK cells. We also observed profound delays in quantitative and functional recovery of viral-specific CD4+ and CD8+ T cell responses for the first year post-CBT. Taken together, our data support efforts aimed at optimizing viral-specific T cell recovery to improve outcomes post-CBT. PMID:25708219

  4. Chromosomal double-strand break repair in Ku80-deficient cells.

    PubMed Central

    Liang, F; Romanienko, P J; Weaver, D T; Jeggo, P A; Jasin, M

    1996-01-01

    The x-ray sensitive hamster cell line xrs-6 is deficient in DNA double-strand break (DSB) repair and exhibits impaired V(D)J recombination. The molecular defect in this line is in the 80-kDa subunit of the Ku autoantigen, a protein that binds to DNA ends and recruits the DNA-dependent protein kinase to DNA. Using an I-SceI endonuclease expression system, chromosomal DSB repair was examined in xrs-6 and parental CHO-K1 cell lines. A DSB in chromosomal DNA increased the yield of recombinants several thousand-fold above background in both the xrs-6 and CHO-K1 cells, with recombinational repair of DSBs occurring in as many as 1 of 100 cells electroporated with the endonuclease expression vector. Thus, recombinational repair of chromosomal DSBs can occur at substantial levels in mammalian cells and it is not grossly affected in our assay by a deficiency of the Ku autoantigen. Rejoining of broken chromosome ends (end-joining) near the site of the DSB was also examined. In contrast to recombinational repair, end-joining was found to be severely impaired in the xrs-6 cells. Thus, the Ku protein appears to play a critical role in only one of the chromosomal DSB repair pathways. Images Fig. 1 Fig. 2 PMID:8799130

  5. Second-order Knudsen-layer analysis for the generalized slip-flow theory: Boundary curvature effects

    NASA Astrophysics Data System (ADS)

    Hattori, Masanari; Takata, Shigeru

    2015-11-01

    A systematic asymptotic analysis of the Boltzmann equation shows that the overall behavior of a gas can be described by fluid-dynamic-type equations with the appropriate slip/jump boundary condition when the Knudsen number is small [the generalized slip-flow theory; see Y. Sone, Molecular Gas Dynamics (Birkhäuser, Boston, 2007)]. Near the boundary, a non-fluid-like correction (the Knudsen-layer correction) to the overall solution is required. Although the theory itself has been established up to the second order of the Knudsen number expansion, the data of the correction have been lacking for a long time for the original Boltzmann equation. Recently, we have obtained the required data, except for the effects of boundary curvature, assuming the hard-sphere molecules and the diffuse reflection boundary condition. In the present work, the effects of boundary curvature have been clarified in details, thereby completing the required numerical data. A local singularity appears at the level of the velocity distribution function. We have developed the numerical method that handles such a singularity safely. The present work is supported in part by KAKENHI from JSPS (Nos. 23360083 and 13J01011).

  6. Vaporisation studies on tellurium dioxide: A Knudsen effusion mass spectrometric study

    NASA Astrophysics Data System (ADS)

    Lakshmi Narasimhan, T. S.; Balasubramanian, R.; Nalini, S.; Sai Baba, M.

    1997-08-01

    The vaporisation of TeO 2(s) was studied by Knudsen effusion mass spectrometry. The vapour phase was found to consist of (TeO 2) n(n = 1-3)(g), (TeO) n(g) (n = 1-3) and Te 2(g). The p- T relations of TeO 2(g), (TeO 2) 2(g) and (TeO 2) 3(g) were derived to be log( p/Pa) = (-13534 ± 78)/ T + (14.241 ± 0.09) (750-950 K), log( p/Pa) = (-14823 ± 212)/T + (14.373 ± 0.242) (825-950 K) and log( p/Pa) = (-19074 ± 540)/ T+ (17.337 ± 0.606) (850-921 K) respectively. From the partial pressures, Δ rH298.150 of nTeO 2(s) = (TeO 2) n(g) ( n = 1-3) were evaluated by second and third law methods. Also, enthalpy of the pressure independent reaction TeO 2(s) + TeO 2(g) = (TeO 2) 2(g) has been evaluated. Using the Δ fH 298.150 of TeO 2(s), Δ fH 298.150 of (Te0 2) n(g) ( n= 1-3) were calculated. The partial pressure and enthalpy data for (TeO 2) 3(g) have been obtained for the first time.

  7. Expressions for the Evaporation and Condensation Coefficients in the Hertz-Knudsen Relation.

    PubMed

    Persad, Aaron H; Ward, Charles A

    2016-07-27

    Although the Hertz-Knudsen (HK) relation is often used to correlate evaporation data, the relation contains two empirical parameters (the evaporation and condensation coefficients) that have inexplicably been found to span 3 orders of magnitude. Explicit expressions for these coefficients have yet to be determined. This review will examine sources of error in the HK relation that have led to the coefficients' scatter. Through an examination of theoretical, experimental, and molecular dynamics simulation studies of evaporation, this review will show that the HK relation is incomplete, since it is missing an important physical concept: the coupling between the vapor and liquid phases during evaporation. The review also examines a modified HK relation, obtained from the quantum-mechanically based statistical rate theory (SRT) expression for the evaporation flux and applying a limit to it in which the thermal energy is dominant. Explicit expressions for the evaporation and condensation coefficients are defined in this limit, with the surprising result that the coefficients are not bounded by unity. An examination is made with 127 reported evaporation experiments of water and of ethanol, leading to a new physical interpretation of the coefficients. The review concludes by showing how seemingly small simplifications, such as assuming thermal equilibrium across the liquid-vapor interface during evaporation, can lead to the erroneous predictions from the HK relation that have been reported in the literature. PMID:27314250

  8. Tail-ion transport and Knudsen layer formation in the presence of magnetic fields

    NASA Astrophysics Data System (ADS)

    Schmit, Paul; Molvig, Kim

    2013-10-01

    The impact of magnetic fields on Knudsen layer formation in ICF-relevant plasma is investigated for the first time. Magnetic fields change the energy scaling of the ion diffusivity in a way that eliminates the preferential losses of fast ions compared to thermal ions. Simple threshold criteria give conditions such that the restoration of the ion tail distribution is sufficient to recover much of the lost fusion reactivity. The tail-ion kinetic equations are solved for hot fuel bounded by a cold, nonreacting wall using a numerical stochastic differential equation solver, and the modified fusion reactivities are calculated. We find that modest magnetic fields too weak to magnetize thermal ions are still sufficient to restore much of the lost reactivity, consistent with the threshold conditions. We also find that the Maxwell-averaged fusion reactivities are recovered more fully in cylindrical targets compared to spherical targets. Sandia National Laboratories is a multi-program laboratory managed and operated by Sandia Corporation, a wholly owned subsidiary of Lockheed Martin Corporation, for the U.S. Department of Energy's National Nuclear Security Administration under contract DE-AC04-94AL85000.

  9. Placement-aware decomposition of a digital standard cells library for double patterning lithography

    NASA Astrophysics Data System (ADS)

    Wassal, Amr G.; Sharaf, Heba; Hammouda, Sherif

    2012-11-01

    To continue scaling the circuit features down, Double Patterning (DP) technology is needed in 22nm technologies and lower. DP requires decomposing the layout features into two masks for pitch relaxation, such that the spacing between any two features on each mask is greater than the minimum allowed mask spacing. The relaxed pitches of each mask are then processed on two separate exposure steps. In many cases, post-layout decomposition fails to decompose the layout into two masks due to the presence of conflicts. Post-layout decomposition of a standard cells block can result in native conflicts inside the cells (internal conflict), or native conflicts on the boundary between two cells (boundary conflict). Resolving native conflicts requires a redesign and/or multiple iterations for the placement and routing phases to get a clean decomposition. Therefore, DP compliance must be considered in earlier phases, before getting the final placed cell block. The main focus of this paper is generating a library of decomposed standard cells to be used in a DP-aware placer. This library should contain all possible decompositions for each standard cell, i.e., these decompositions consider all possible combinations of boundary conditions. However, the large number of combinations of boundary conditions for each standard cell will significantly increase the processing time and effort required to obtain all possible decompositions. Therefore, an efficient methodology is required to reduce this large number of combinations. In this paper, three different reduction methodologies are proposed to reduce the number of different combinations processed to get the decomposed library. Experimental results show a significant reduction in the number of combinations and decompositions needed for the library processing. To generate and verify the proposed flow and methodologies, a prototype for a placement-aware DP-ready cell-library is developed with an optimized number of cell views.

  10. Zebrafish Class 1 Phosphatidylinositol Transfer Proteins: PITPβ and Double Cone Cell Outer Segment Integrity in Retina

    PubMed Central

    Ile, Kristina E.; Kassen, Sean; Cao, Canhong; Vihtehlic, Thomas; Shah, Sweety D.; Mousley, Carl J.; Alb, James G.; Huijbregts, Richard P.H.; Stearns, George W.; Brockerhoff, Susan E.; Hyde, David R.; Bankaitis, Vytas A.

    2010-01-01

    Phosphatidylinositol transfer proteins (PITPs) in yeast coordinate lipid metabolism with the activities of specific membrane trafficking pathways. The structurally unrelated metazoan-specific PITPs (mPITPs), on the other hand, are an under-investigated class of proteins. It remains unclear what biological activities mPITPs discharge, and the mechanisms by which these proteins function are also not understood. The soluble class 1 mPITPs include the PITPα and PITPβ isoforms. Of these, the β-isoforms are particularly poorly characterized. Herein, we report the use of zebrafish as a model vertebrate for the study of class 1 mPITP biological function. Zebrafish express PITPα and PITPβ-isoforms (Pitpna and Pitpnb, respectively) and a novel PITPβ-like isoform (Pitpng). Pitpnb expression is particularly robust in double cone cells of the zebrafish retina. Morpholino-mediated protein knockdown experiments demonstrate Pitpnb activity is primarily required for biogenesis/maintenance of the double cone photoreceptor cell outer segments in the developing retina. By contrast, Pitpna activity is essential for successful navigation of early developmental programs. This study reports the initial description of the zebrafish class 1 mPITP family, and the first analysis of PITPβ function in a vertebrate. PMID:20545905

  11. [Establishment of lentivirus-mediated system of double suicide genes and its killing effects on K562 cells].

    PubMed

    Jiang, Yi-Rong; Liu, Chun-Sheng; Chen, Xue-Liang; Ma, Dao-Xin

    2004-02-01

    To establish lentivirus-mediated system of double suicide genes and explore its killing effects on K562 cells, lentivirus transfer vector for double suicide genes was constructed using molecular methods, three plasmids of lentivirus gene transfer vector system were transferred into packaging cell line 293T using lipofectine method, the transfer effect was observed through fluorescence microscopy, the lentivirus particles were observed by means of electron microscopy. High titer of lentivirus was harvested from the supernatant of virus-producing cell culture and concentrated by high-speed centrifugation with Poly-L-Lysine (PLL). The K562 cells were infected with the concentrated supernatant containing the virus with the double suicide genes. Fluorescence microscopy and RT- PCR confirmed the integration and expression of extraneous gene. The cytotoxicity to these transgenic cells treated with 5-FC and GCV was measured by MTT assays. The growth inhibition ratio (GIR) of cells and inhibition concentration 50 (IC(50)) were counted. After administration of GCV and 5-FC, the changes of those cells were observed through scanning electron microscope. The results showed that lentivirus transfer vector with double suicide genes was constructed successfully. The above-mentioned plasmids were effectively transferred into 293T cells. So much green fluorescence was observed through fluorescence microscope. A lot of lentivirus particles were observed through transmission electron microscope. Double suicide genes mediated by lentivirus were stably integrated and expressed in K562 cells after infection with the concentrated virus using fluorescence microscopy and RT-PCR. The GIR of K562 cells using GCV or 5-FC was 48.73% or 50.69% respectively and it was apparently higher than that of untransfected cells (P < 0.01). When using GCV and 5-FC together, the GIR was 87.69%, which was apparently higher than that of group using GCV or 5-FC alone (P < 0.01). In conclusion, lentivirus

  12. Single-cell microarray enables high-throughput evaluation of DNA double-strand breaks and DNA repair inhibitors.

    PubMed

    Weingeist, David M; Ge, Jing; Wood, David K; Mutamba, James T; Huang, Qiuying; Rowland, Elizabeth A; Yaffe, Michael B; Floyd, Scott; Engelward, Bevin P

    2013-03-15

    A key modality of non-surgical cancer management is DNA damaging therapy that causes DNA double-strand breaks that are preferentially toxic to rapidly dividing cancer cells. Double-strand break repair capacity is recognized as an important mechanism in drug resistance and is therefore a potential target for adjuvant chemotherapy. Additionally, spontaneous and environmentally induced DSBs are known to promote cancer, making DSB evaluation important as a tool in epidemiology, clinical evaluation and in the development of novel pharmaceuticals. Currently available assays to detect double-strand breaks are limited in throughput and specificity and offer minimal information concerning the kinetics of repair. Here, we present the CometChip, a 96-well platform that enables assessment of double-strand break levels and repair capacity of multiple cell types and conditions in parallel and integrates with standard high-throughput screening and analysis technologies. We demonstrate the ability to detect multiple genetic deficiencies in double-strand break repair and evaluate a set of clinically relevant chemical inhibitors of one of the major double-strand break repair pathways, non-homologous end-joining. While other high-throughput repair assays measure residual damage or indirect markers of damage, the CometChip detects physical double-strand breaks, providing direct measurement of damage induction and repair capacity, which may be useful in developing and implementing treatment strategies with reduced side effects. PMID:23422001

  13. Single-cell microarray enables high-throughput evaluation of DNA double-strand breaks and DNA repair inhibitors

    PubMed Central

    Weingeist, David M.; Ge, Jing; Wood, David K.; Mutamba, James T.; Huang, Qiuying; Rowland, Elizabeth A.; Yaffe, Michael B.; Floyd, Scott; Engelward, Bevin P.

    2013-01-01

    A key modality of non-surgical cancer management is DNA damaging therapy that causes DNA double-strand breaks that are preferentially toxic to rapidly dividing cancer cells. Double-strand break repair capacity is recognized as an important mechanism in drug resistance and is therefore a potential target for adjuvant chemotherapy. Additionally, spontaneous and environmentally induced DSBs are known to promote cancer, making DSB evaluation important as a tool in epidemiology, clinical evaluation and in the development of novel pharmaceuticals. Currently available assays to detect double-strand breaks are limited in throughput and specificity and offer minimal information concerning the kinetics of repair. Here, we present the CometChip, a 96-well platform that enables assessment of double-strand break levels and repair capacity of multiple cell types and conditions in parallel and integrates with standard high-throughput screening and analysis technologies. We demonstrate the ability to detect multiple genetic deficiencies in double-strand break repair and evaluate a set of clinically relevant chemical inhibitors of one of the major double-strand break repair pathways, non-homologous end-joining. While other high-throughput repair assays measure residual damage or indirect markers of damage, the CometChip detects physical double-strand breaks, providing direct measurement of damage induction and repair capacity, which may be useful in developing and implementing treatment strategies with reduced side effects. PMID:23422001

  14. Imitation Monte Carlo methods for problems of the Boltzmann equation with small Knudsen numbers, parallelizing algorithms with splitting

    NASA Astrophysics Data System (ADS)

    Khisamutdinov, A. I.; Velker, N. N.

    2014-05-01

    The talk examines a system of pairwise interaction particles, which models a rarefied gas in accordance with the nonlinear Boltzmann equation, the master equations of Markov evolution of this system and corresponding numerical Monte Carlo methods. Selection of some optimal method for simulation of rarefied gas dynamics depends on the spatial size of the gas flow domain. For problems with the Knudsen number Kn of order unity "imitation", or "continuous time", Monte Carlo methods ([2]) are quite adequate and competitive. However if Kn <= 0.1 (the large sizes), excessive punctuality, namely, the need to see all the pairs of particles in the latter, leads to a significant increase in computational cost(complexity). We are interested in to construct the optimal methods for Boltzmann equation problems with large enough spatial sizes of the flow. Speaking of the optimal, we mean that we are talking about algorithms for parallel computation to be implemented on high-performance multi-processor computers. The characteristic property of large systems is the weak dependence of sub-parts of each other at a sufficiently small time intervals. This property is taken into account in the approximate methods using various splittings of operator of corresponding master equations. In the paper, we develop the approximate method based on the splitting of the operator of master equations system "over groups of particles" ([7]). The essence of the method is that the system of particles is divided into spatial subparts which are modeled independently for small intervals of time, using the precise"imitation" method. The type of splitting used is different from other well-known type "over collisions and displacements", which is an attribute of the known Direct simulation Monte Carlo methods. The second attribute of the last ones is the grid of the "interaction cells", which is completely absent in the imitation methods. The main of talk is parallelization of the imitation algorithms with

  15. Repair of Chromosomal Double-Strand Breaks by Precise Ligation in Human Cells

    PubMed Central

    Lin, William Y.; Wilson, John H.; Lin, Yunfu

    2013-01-01

    Double-strand breaks (DSBs), a common type of DNA lesion, occur daily in human cells as a result of both endogenous and exogenous damaging agents. DSBs are repaired in two general ways: by the homology-dependent, error-free pathways of homologous recombination (HR) and by the homology-independent, error-prone pathways of nonhomologous end-joining (NHEJ), with NHEJ predominating in most cells. DSBs with compatible ends can be re-joined in vitro with DNA ligase alone, which raises the question of whether such DSBs require the more elaborate machinery of NHEJ to be repaired in cells. Here we report that chromosomal DSBs with compatible ends introduced by the rare-cutting endonuclease, ISceI, are repaired by precise ligation nearly 100% of the time in human cells. Precise ligation depends on the classical NHEJ components Ku70, XRCC4, and DNA ligase IV, since siRNA knockdowns of these factors significantly reduced the efficiency of precise ligation. Interestingly, knockdown of the tumor suppressors p53 or BRCA1 showed similar effects as the knockdowns of NHEJ factors. In contrast, knockdown of components involved in alternative NHEJ, mismatch repair, nucleotide excision repair, and single-strand break repair did not reduce precise ligation. In summary, our results demonstrate that DSBs in human cells are efficiently repaired by precise ligation, which requires classical NHEJ components and is enhanced by p53 and BRCA1. PMID:23707303

  16. Measuring electrical and mechanical properties of red blood cells with a double optical tweezers

    NASA Astrophysics Data System (ADS)

    Fontes, Adriana; Fernandes, Heloise P.; Barjas-Castro, Maria L.; de Thomaz, André A.; Pozzo, Liliana d. Y.; Barbosa, Luiz C.; Cesar, Carlos L.

    2006-08-01

    The fluid lipid bilayer viscoelastic membrane of red blood cells (RBC) contains antigen glycolproteins and proteins which can interact with antibodies to cause cell agglutination. This is the basis of most of the immunohematologic tests in blood banks and the identification of the antibodies against the erythrocyte antigens is of fundamental importance for transfusional routines. The negative charges of the RBCs creates a repulsive electric (zeta) potential between the cells and prevents their aggregation in the blood stream. The first counterions cloud strongly binded moving together with the RBC is called the compact layer. This report proposes the use of a double optical tweezers for a new procedure for measuring: (1) the apparent membrane viscosity, (2) the cell adhesion, (3) the zeta potential and (4) the compact layer's size of the charges formed around the cell in the electrolytic solution. To measure the membrane viscosity we trapped silica beads strongly attached to agglutinated RBCs and measured the force to slide one RBC over the other as a function of the relative velocity. The RBC adhesion was measured by slowly displacing two RBCs apart until the disagglutination happens. The compact layer's size was measured using the force on the silica bead attached to a single RBC in response to an applied voltage and the zeta potential was obtained by measuring the terminal velocity after releasing the RBC from the optical trap at the last applied voltage. We believe that the methodology here proposed can improve the methods of diagnosis in blood banks.

  17. DNA double strand breaks and Hsp70 expression in proton irradiated living cells

    NASA Astrophysics Data System (ADS)

    Fiedler, Anja; Reinert, Tilo; Tanner, Judith; Butz, Tilman

    2007-07-01

    DNA double strand breaks (DSBs) in living cells can be directly provoked by ionising radiation. DSBs can be visualized by immunostaining the phosphorylated histone γH2AX. Our concern was to test the feasibility of γH2AX staining for a direct visualization of single proton hits. If single protons produce detectable foci, DNA DSBs could be used as "biological track detectors" for protons. Ionising radiation can also damage proteins indirectly by inducing free radicals. Heat shock proteins (Hsp) help to refold or even degrade the damaged proteins. The level of the most famous heat shock protein Hsp70 is increased by ionising radiation. We investigated the expression of γH2AX and Hsp70 after cross and line patterned irradiation with counted numbers of 2.25 MeV protons on primary human skin fibroblasts. The proton induced DSBs appear more delocalised than it was expected by the ion hit accuracy. Cooling the cells before the irradiation reduces the delocalisation of DNA DSBs, which is probably caused by the reduced diffusion of DNA damaging agents. Proton irradiation seems to provoke protein damages mainly in the cytoplasm indicated by cytoplasmic Hsp70 aggregates. On the contrary, in control heat shocked cells the Hsp70 was predominantly localized in the cell nucleus. However, the irradiated area could not be recognized, all cells on the Si 3N 4 window showed a homogenous Hsp70 expression pattern.

  18. TiO2 nanoparticles induce DNA double strand breaks and cell cycle arrest in human alveolar cells.

    PubMed

    Kansara, Krupa; Patel, Pal; Shah, Darshini; Shukla, Ritesh K; Singh, Sanjay; Kumar, Ashutosh; Dhawan, Alok

    2015-03-01

    TiO2 nanoparticles (NPs) have the second highest global annual production (∼3000 tons) among the metal-containing NPs. These NPs are used as photocatalysts for bacterial disinfection, and in various other consumer products including sunscreen, food packaging, therapeutics, biosensors, surface cleaning agents, and others. Humans are exposed to these NPs during synthesis (laboratory), manufacture (industry), and use (consumer products, devices, medicines, etc.), as well as through environmental exposures (disposal). Hence, there is great concern regarding the health effects caused by exposure to NPs and, in particular, to TiO2 NPs. In the present study, the genotoxic potential of TiO2 NPs in A549 cells was examined, focusing on their potential to induce ROS, different types of DNA damage, and cell cycle arrest. We show that TiO2 NPs can induce DNA damage and a corresponding increase in micronucleus frequency, as evident from the comet and cytokinesis-block micronucleus assays. We demonstrate that DNA damage may be attributed to increased oxidative stress and ROS generation. Furthermore, genomic and proteomic analyses showed increased expression of ATM, P53, and CdC-2 and decreased expression of ATR, H2AX, and Cyclin B1 in A549 cells, suggesting induction of DNA double strand breaks. The occurrence of double strand breaks was correlated with cell cycle arrest in G2/M phase. Overall, the results indicate the potential for genotoxicity following exposure to these TiO2 NPs, suggesting that use should be carefully monitored. PMID:25524809

  19. Layered Double Hydroxide Nanotransporter for Molecule Delivery to Intact Plant Cells.

    PubMed

    Bao, Wenlong; Wang, Junya; Wang, Qiang; O'Hare, Dermot; Wan, Yinglang

    2016-01-01

    Here we report a powerful method that facilitates the transport of biologically active materials across the cell wall barrier in plant cells. Positively charged delaminated layered double hydroxide lactate nanosheets (LDH-lactate-NS) with a 0.5‒2 nm thickness and 30‒60 nm diameter exhibit a high adsorptive capacity for negatively charged biomolecules, including fluorescent dyes such as tetramethyl rhodamine isothiocyanate (TRITC), fluorescein isothiocyanate isomer I(FITC) and DNA molecules, forming neutral LDH-nanosheet conjugates. These neutral conjugates can shuttle the bound fluorescent dye into the cytosol of intact plant cell very efficiently. Furthermore, typical inhibitors of endocytosis and low temperature incubation did not prevent LDH-lactate-NS internalization, suggesting that LDH-lactate-NS penetrated the plasma membrane via non-endocytic pathways, which will widen the applicability to a variety of plant cells. Moreover, the absence of unwanted side effects in our cytological studies, and the nuclear localization of ssDNA-FITC suggest that nano-LDHs have potential application as a novel gene carrier to plants. PMID:27221055

  20. Layered Double Hydroxide Nanotransporter for Molecule Delivery to Intact Plant Cells

    PubMed Central

    Bao, Wenlong; Wang, Junya; Wang, Qiang; O’Hare, Dermot; Wan, Yinglang

    2016-01-01

    Here we report a powerful method that facilitates the transport of biologically active materials across the cell wall barrier in plant cells. Positively charged delaminated layered double hydroxide lactate nanosheets (LDH-lactate-NS) with a 0.5‒2 nm thickness and 30‒60 nm diameter exhibit a high adsorptive capacity for negatively charged biomolecules, including fluorescent dyes such as tetramethyl rhodamine isothiocyanate (TRITC), fluorescein isothiocyanate isomer I(FITC) and DNA molecules, forming neutral LDH-nanosheet conjugates. These neutral conjugates can shuttle the bound fluorescent dye into the cytosol of intact plant cell very efficiently. Furthermore, typical inhibitors of endocytosis and low temperature incubation did not prevent LDH-lactate-NS internalization, suggesting that LDH-lactate-NS penetrated the plasma membrane via non-endocytic pathways, which will widen the applicability to a variety of plant cells. Moreover, the absence of unwanted side effects in our cytological studies, and the nuclear localization of ssDNA-FITC suggest that nano-LDHs have potential application as a novel gene carrier to plants. PMID:27221055

  1. Using carbon nanotubes to induce micronuclei and double strand breaks of the DNA in human cells

    NASA Astrophysics Data System (ADS)

    Cveticanin, Jelena; Joksic, Gordana; Leskovac, Andreja; Petrovic, Sandra; Valenta Sobot, Ana; Neskovic, Olivera

    2010-01-01

    Carbon nanotubes are unique one-dimensional macromolecules with promising applications in biology and medicine. Since their toxicity is still under debate, here we present a study investigating the genotoxic properties of purified single wall carbon nanotubes (SWCNTs), multiwall carbon nanotubes (MWCNTs), and amide functionalized purified SWCNTs on cultured human lymphocytes employing cytokinesis block micronucleus assay and enumeration of γH2AX foci as a measure of double strand breaks (DSBs) of the DNA in normal human fibroblasts. SWCNTs induce micronuclei (MN) formation in lymphocytes and decrease the proliferation potential (CBPI) of cells. In a fibroblast cell line the same dose of SWCNTs induces γH2AX foci 2.7-fold higher than in a control. Amide functionalized purified SWCNTs behave differently: they do not disturb the cell proliferation potential of harvested lymphocytes, but induce micronuclei to a higher extent than SWCNTs. When applied on fibroblasts, amide functionalized SWCNTs also induce γH2AX foci, 3.18-fold higher than the control. The cellular effects of MWCNTs display the broad spectrum of clastogenic properties seen as the highest incidence of induced lymphocyte micronuclei and anaphase bridges among nuclei in binucleated cells. Surprisingly, the incidence of induced γH2AX foci was not as high as was expected by the micronucleus test, which indicates that MWCNTs act as clastogen and aneugen agents simultaneously. Biological endpoints investigated in this study indicate a close relationship between the electrochemical properties of carbon nanotubes and observed genotoxicity.

  2. Sulforaphane induces DNA double strand breaks predominantly repaired by homologous recombination pathway in human cancer cells

    SciTech Connect

    Sekine-Suzuki, Emiko; Yu, Dong; Kubota, Nobuo; Okayasu, Ryuichi; Anzai, Kazunori

    2008-12-12

    Cytotoxicity and DNA double strand breaks (DSBs) were studied in HeLa cells treated with sulforaphane (SFN), a well-known chemo-preventive agent. Cell survival was impaired by SFN in a concentration and treatment time-dependent manner. Both constant field gel electrophoresis (CFGE) and {gamma}-H2AX assay unambiguously indicated formation of DSBs by SFN, reflecting the cell survival data. These DSBs were predominantly processed by homologous recombination repair (HRR), judging from the SFN concentration-dependent manner of Rad51 foci formation. On the other hand, the phosphorylation of DNA-PKcs, a key non-homologous end joining (NHEJ) protein, was not observed by SFN treatment, suggesting that NHEJ may not be involved in DSBs induced by this chemical. G2/M arrest by SFN, a typical response for cells exposed to ionizing radiation was also observed. Our new data indicate the clear induction of DSBs by SFN and a useful anti-tumor aspect of SFN through the induction of DNA DSBs.

  3. Using carbon nanotubes to induce micronuclei and double strand breaks of the DNA in human cells.

    PubMed

    Cveticanin, Jelena; Joksic, Gordana; Leskovac, Andreja; Petrovic, Sandra; Sobot, Ana Valenta; Neskovic, Olivera

    2010-01-01

    Carbon nanotubes are unique one-dimensional macromolecules with promising applications in biology and medicine. Since their toxicity is still under debate, here we present a study investigating the genotoxic properties of purified single wall carbon nanotubes (SWCNTs), multiwall carbon nanotubes (MWCNTs), and amide functionalized purified SWCNTs on cultured human lymphocytes employing cytokinesis block micronucleus assay and enumeration of gamma H2AX foci as a measure of double strand breaks (DSBs) of the DNA in normal human fibroblasts. SWCNTs induce micronuclei (MN) formation in lymphocytes and decrease the proliferation potential (CBPI) of cells. In a fibroblast cell line the same dose of SWCNTs induces gamma H2AX foci 2.7-fold higher than in a control. Amide functionalized purified SWCNTs behave differently: they do not disturb the cell proliferation potential of harvested lymphocytes, but induce micronuclei to a higher extent than SWCNTs. When applied on fibroblasts, amide functionalized SWCNTs also induce gamma H2AX foci, 3.18-fold higher than the control. The cellular effects of MWCNTs display the broad spectrum of clastogenic properties seen as the highest incidence of induced lymphocyte micronuclei and anaphase bridges among nuclei in binucleated cells. Surprisingly, the incidence of induced gamma H2AX foci was not as high as was expected by the micronucleus test, which indicates that MWCNTs act as clastogen and aneugen agents simultaneously. Biological endpoints investigated in this study indicate a close relationship between the electrochemical properties of carbon nanotubes and observed genotoxicity. PMID:19946169

  4. Radiation-induced DNA double-strand break rejoining in human tumour cells.

    PubMed Central

    Núñez, M. I.; Villalobos, M.; Olea, N.; Valenzuela, M. T.; Pedraza, V.; McMillan, T. J.; Ruiz de Almodóvar, J. M.

    1995-01-01

    Five established human breast cancer cell lines and one established human bladder cancer cell line of varying radiosensitivity have been used to determine whether the rejoining of DNA double-strand breaks (dsbs) shows a correlation with radiosensitivity. The kinetics of dsb rejoining was biphasic and both components proceeded exponentially with time. The half-time (t1/2) of rejoining ranged from 18.0 +/- 1.4 to 36.4 +/- 3.2 min (fast rejoining process) and from 1.5 +/- 0.2 to 5.1 +/- 0.2 h (slow rejoining process). We found a statistically significant relationship between the survival fraction at 2 Gy (SF2) and the t1/2 of the fast rejoining component (r = 0.949, P = 0.0039). Our results suggest that cell lines which show rapid rejoining are more radioresistant. These results support the view that, as well as the level of damage induction that we have reported previously, the repair process is a major determinant of cellular radiosensitivity. It is possible that the differences found in DNA dsb rejoining and the differences in DNA dsb induction are related by a common mechanism, e.g. conformation of chromatin in the cell. PMID:7841046

  5. RNF4 regulates DNA double-strand break repair in a cell cycle-dependent manner

    PubMed Central

    Kuo, Ching-Ying; Li, Xu; Stark, Jeremy M.; Shih, Hsiu-Ming; Ann, David K.

    2016-01-01

    Abstract Both RNF4 and KAP1 play critical roles in the response to DNA double-strand breaks (DSBs), but the functional interplay of RNF4 and KAP1 in regulating DNA damage response remains unclear. We have previously demonstrated the recruitment and degradation of KAP1 by RNF4 require the phosphorylation of Ser824 (pS824) and SUMOylation of KAP1. In this report, we show the retention of DSB-induced pS824-KAP1 foci and RNF4 abundance are inversely correlated as cell cycle progresses. Following irradiation, pS824-KAP1 foci predominantly appear in the cyclin A (-) cells, whereas RNF4 level is suppressed in the G0-/G1-phases and then accumulates during S-/G2-phases. Notably, 53BP1 foci, but not BRCA1 foci, co-exist with pS824-KAP1 foci. Depletion of KAP1 yields opposite effect on the dynamics of 53BP1 and BRCA1 loading, favoring homologous recombination repair. In addition, we identify p97 is present in the RNF4-KAP1 interacting complex and the inhibition of p97 renders MCF7 breast cancer cells relatively more sensitive to DNA damage. Collectively, these findings suggest that combined effect of dynamic recruitment of RNF4 to KAP1 regulates the relative occupancy of 53BP1 and BRCA1 at DSB sites to direct DSB repair in a cell cycle-dependent manner. PMID:26766492

  6. RNF4 regulates DNA double-strand break repair in a cell cycle-dependent manner.

    PubMed

    Kuo, Ching-Ying; Li, Xu; Stark, Jeremy M; Shih, Hsiu-Ming; Ann, David K

    2016-03-18

    Both RNF4 and KAP1 play critical roles in the response to DNA double-strand breaks (DSBs), but the functional interplay of RNF4 and KAP1 in regulating DNA damage response remains unclear. We have previously demonstrated the recruitment and degradation of KAP1 by RNF4 require the phosphorylation of Ser824 (pS824) and SUMOylation of KAP1. In this report, we show the retention of DSB-induced pS824-KAP1 foci and RNF4 abundance are inversely correlated as cell cycle progresses. Following irradiation, pS824-KAP1 foci predominantly appear in the cyclin A (-) cells, whereas RNF4 level is suppressed in the G0-/G1-phases and then accumulates during S-/G2-phases. Notably, 53BP1 foci, but not BRCA1 foci, co-exist with pS824-KAP1 foci. Depletion of KAP1 yields opposite effect on the dynamics of 53BP1 and BRCA1 loading, favoring homologous recombination repair. In addition, we identify p97 is present in the RNF4-KAP1 interacting complex and the inhibition of p97 renders MCF7 breast cancer cells relatively more sensitive to DNA damage. Collectively, these findings suggest that combined effect of dynamic recruitment of RNF4 to KAP1 regulates the relative occupancy of 53BP1 and BRCA1 at DSB sites to direct DSB repair in a cell cycle-dependent manner. PMID:26766492

  7. Characteristics of bladder transitional cell carcinoma with E-cadherin and N-cadherin double-negative expression

    PubMed Central

    LUO, YANG; ZHU, YONG-TONG; MA, LI-LI; PANG, SHI-YU; WEI, LI-JIE; LEI, CHENG-YONG; HE, CHENG-WU; TAN, WAN-LONG

    2016-01-01

    The aim of the present study was to examine the characteristics of bladder transitional cell carcinoma with E-cadherin and N-cadherin double-negative expression. An immunofluorescence assay was used to detect E-cadherin and N-cadherin expression in infiltrative bladder cancer tissues, and immunofluorescence and western blot analysis were used to detect E-cadherin and N-cadherin expression in human urinary bladder grade II carcinoma 5637, transitional cell carcinoma UMUC-3 and invasive bladder carcinoma EJ cells. Cell proliferation, migration, invasion and plate colony formation assays were used to detect the proliferative, migratory and invasive abilities and the efficiency of plate colony formation of 5637, UMUC3 and EJ cells. A tumor xenograft formation assay was used to evaluate the tumorigenic abilities of 5637, UMUC-3 and EJ cells in vivo. E-cadherin and N-cadherin double-negative expression was identified in various pathological grades of infiltrative bladder cancers. E-cadherin positive and N-cadherin negative expression was exhibited by 5637 cells. By contrast, E-cadherin negative and N-cadherin positive expression was exhibited by EJ cells, and E-cadherin and N-cadherin double-negative expression was exhibited by UMUC-3 cells. The ability of cells to proliferate, migrate, invade, and the efficiency of plate colony formation and tumorigenic abilities of the cells were significantly different among 5637, UMUC-3 and EJ cells. These cell characteristics were significantly increased in UMUC-3 cells compared with 5637 cells; however, the characteristics were significantly decreased compared with EJ cells. The biological characteristics of bladder cancer cells with E-cadherin and N-cadherin double-negative expression was between bladder cancer cells that exhibited a E-cadherin positive and N-cadherin negative expression, and bladder cancer cells that exhibited E-cadherin negative and N-cadherin positive expression. The present study deduces that the status of E

  8. CHO cell repair of single-strand and double-strand DNA breaks induced by gamma- and alpha-radiations.

    PubMed

    Cole, A; Shonka, F; Corry, P; Cooper, W G

    1975-01-01

    Neutral and alkaline sucrose gradient sedimentation analysis was used to measure double- and single-strand breaks in the DNA of Chinese hamster ovary (CHO) cells exposed to either gamma- or alpha-radiation. After irradiation, cells were incubated for 15-180 min to test the ability of the cell to rejoin the DNA breaks. Essentially complete rejoining was observed for single-strand breaks induced by gamma- or alpha-doses below 20 krad and for double-strand breaks induced by gamma doses below 60 krad. Approximately 80% rejoining was observed for double-strand breaks induced by alpha doses below 40 krad. At higher doses, the repair system appeared to saturate in such a way that essentially no additional breaks were rejoined. PMID:1191188

  9. MeHg Developing Exposure Causes DNA Double-Strand Breaks and Elicits Cell Cycle Arrest in Spinal Cord Cells

    PubMed Central

    Ferreira, Fabiana F.; Ammar, Dib; Bourckhardt, Gilian F.; Kobus-Bianchini, Karoline; Müller, Yara M. R.; Nazari, Evelise M.

    2015-01-01

    The neurotoxicity caused by methylmercury (MeHg) is well documented; however, the developmental neurotoxicity in spinal cord is still not fully understood. Here we investigated whether MeHg affects the spinal cord layers development. Chicken embryos at E3 were treated in ovo with 0.1 μg MeHg/50 μL saline solution and analyzed at E10. Thus, we performed immunostaining using anti-γ-H2A.X to recognize DNA double-strand breaks and antiphosphohistone H3, anti-p21, and anti-cyclin E to identify cells in proliferation and cell cycle proteins. Also, to identify neuronal cells, we used anti-NeuN and anti-βIII-tubulin antibodies. After the MeHg treatment, we observed the increase on γ-H2A.X in response to DNA damage. MeHg caused a decrease in the proliferating cells and in the thickness of spinal cord layers. Moreover, we verified that MeHg induced an increase in the number of p21-positive cells but did not change the cyclin E-positive cells. A significantly high number of TUNEL-positive cells indicating DNA fragmentation were observed in MeHg-treated embryos. Regarding the neuronal differentiation, MeHg induced a decrease in NeuN expression and did not change the expression of βIII-tubulin. These results showed that in ovo MeHg exposure alters spinal cord development by disturbing the cell proliferation and death, also interfering in early neuronal differentiation. PMID:26793240

  10. Component Activity Measurements in the Ti-Al-O System by Knudsen Cell Mass Spectrometry

    NASA Technical Reports Server (NTRS)

    Copland, Evan; Jacobson, Nathan S.

    2003-01-01

    Titanium-aluminides (containing (alpha)2-Ti3Al and gamma-TiAl intermetallic phases) have received continued research focus due to their potential as low-density materials for structural applications at intermediate temperatures. However their application above about 850C is hindered by poor oxidation resistance, characterized by the formation of a non-protective TiO2+Al2O3 scale and an oxygen-enriched subsurface zone. Consistent with this are measured titanium and aluminum activities in "oxygen-free" titanium-aluminides, which indicate Al2O3 is only stable for aluminum concentrations greater then 54 atom percent at 1373 K. However, the inability to form a protective Al2O3 scale is in apparent conflict with phase diagram studies, as experimental isothermal sections of the Ti-Al-O system show gamma-TiAl + alpha2-Ti3Al structures are in equilibrium only with Al2O3. The apparent resolution to this conflict lies in the inclusion of oxygen effects in the thermodynamic measurements

  11. Injection of metallic elements into an electron-beam ion trap using a Knudsen cell

    SciTech Connect

    Yamada, C.; Nagata, K.; Nakamura, N.; Ohtani, S.; Takahashi, S.; Tobiyama, T.; Tona, M.; Watanabe, H.; Yoshiyasu, N.; Sakurai, M.; Kavanagh, A. P.; Currell, F. J.

    2006-06-15

    A method of injecting metallic elements into an electron-beam ion trap (EBIT) is described. The method is advantageous over the conventional coaxial and pulsed injection methods in two ways: (a) complicated switching of injection and extraction beams can be avoided when extracting beams of highly charged ions from the EBIT and (b) a beam of stable intensity can be achieved. This method may be applicable to any metallic elements or metallic compounds that have vapor pressures of {approx}0.1 Pa at a temperature lower than 1900 deg. C. We have employed this method for the extraction of highly charged ions of Bi, Er, Fe, and Ho.

  12. Cultured mucosal cell sheet with a double layer of keratinocytes and fibroblasts on a collagen membrane.

    PubMed

    Imaizumi, Fumiko; Asahina, Izumi; Moriyama, Takashi; Ishii, Masatoshi; Omura, Ken

    2004-01-01

    The aim of this study was to develop a novel cultured mucosal membrane that was facile to prepare and easy to handle, and that could be applied to mucosal defects in the oral cavity. Human oral keratinocytes and fibroblasts were prepared from the oral mucosa. We made the following two types of cultured mucosal cell sheets: a monolayer sheet of keratinocytes cultured on a collagen membrane (K-S) and a double-layered sheet of keratinocytes and fibroblasts on a collagen membrane (KF-S). A collagen membrane was used as a control. Each type of sheet was transplanted onto dorsal skin defects of nude mice. The wound area was measured for the assessment of wound contraction and a specimen was harvested for histologic evaluation 1 week and 4 weeks after grafting. Wound contraction was minimal with KF-S grafts. Although histologic examination showed normal differentiation of the epithelium in all graft types, the involucrin expression pattern of KFS was most similar to that of normal epithelium. These results indicate that a double-layered sheet of keratinocytes and fibroblasts cultured on a collagen membrane may facilitate epithelial healing and prevent wound contraction. PMID:15265283

  13. Study of vaporization of sodium metaborate by transpiration thermogravimetry and Knudsen effusion mass spectrometry.

    PubMed

    Narasimhan, T S Lakshmi; Viswanathan, R; Nalini, S

    2011-11-17

    The vaporization of solid sodium metaborate NaBO(2)(s) was studied by transpiration thermogravimetry (TTG) and Knudsen effusion mass spectrometry (KEMS). The transpiration measurements, performed for the first time on NaBO(2)(s), involved use of argon as the carrier gas for vapor transport and derivation of vapor pressure of NaBO(2)(g) (by assuming it as the sole vapor species) through many flow-dependence runs and temperature-dependence runs in the temperature range 1075-1218 K. The KEMS measurements performed in the temperature range 1060-1185 K confirmed NaBO(2)(g) as the principal vapor species over NaBO(2)(s), in accord with the previously reported KEMS studies. The values of p(NaBO(2)) obtained by both TTG and KEMS are consistent within the uncertainties associated with each method and so are the second- and third-law values of enthalpy of sublimation, the latter aspect consistently missing in all previous vaporization studies. The results of both TTG and KEMS were combined to recommend the following thermodynamic parameters pertinent to the sublimation reaction, NaBO(2)(s) = NaBO(2)(g): Log{p(NaBO(2))/Pa} = -(17056 ± 441)/(T/K) + (14.73 ± 0.35) for the temperature range 1060-1218 K; Δ(r)H°(m)(298.15 K) = (346.3 ± 9.4) kJ·mol(-1); and Δ(r)S°(m)(298.15 K) = (210.2 ± 6.8) J·mol(-1)·K(-1). PMID:21985472

  14. Application of Lithium Attachment Mass Spectrometry for Knudsen Evaporation and Chemical Ionisation Mass Spectrometry (KEMS, CIMS)

    NASA Astrophysics Data System (ADS)

    Bannan, T.; Booth, M.; Benyezzar, M.; Bacak, A.; Alfarra, M. R. R.; Topping, D. O.; Percival, C.

    2015-12-01

    Lithium ion attachment mass spectrometry provides a non-specific, non-fragmenting and sensitive method for detection of volatile species in the gas phase. The design, manufacture, and results from lithium ion attachment ionisation sources for two mass spectrometry systems are presented. Trace gas analysis is investigated using a modified Chemical Ionization Mass Spectrometer (CIMS) and vapour pressure (VP) measurements using a modified Knudsen Effusion Mass Spectrometer (KEMS) are presented. The Li+ modified CIMS provided limits of detection of 4 ppt for acetone, 0.2 ppt for formic acid, 15 ppt for nitric acid and 120 ppt from ammonia. Despite improvements, the problem of burnout remained persistent. The Li+ CIMS would unlikely be suitable for field or aircraft work, but could be appropriate for certain lab applications. The KEMS currently utilizes an electron impact (EI) ionisation source which provides a highly sensitive source, with the drawback of fragmentation of ionized molecules (Booth et al., 2009). Using Li+ KEMS the VP of samples can be measured without fragmentation and can therefore be used to identify VPs of individual components in mixtures. The validity of using Li+ for determining the VP of mixtures was tested by making single component VP measurements, which showed good agreement with EI measurements of Poly ethylene glycol (PEG) 3 and PEG 4, both when individually measured and when mixed. The Li+ KEMS was then used to investigate a system of atmospheric relevance, α-pinene secondary organic aerosol, generated in a reaction chamber (Alfarra et al., 2012). The VPs of the individual components from this generated sample are within the range we expect for compounds capable of partitioning between the particle and gas phase of an aerosol (0.1-10-5 Pa). Li+ source has a calculated sensitivity approximately 75 times less than that of EI, but the lack of fragmentation using the Li+ source is a significant advantage.

  15. Application of Lithium Attachment Mass Spectrometry for Knudsen Evaporation and Chemical Ionisation Mass Spectrometry (KEMS, CIMS)

    NASA Astrophysics Data System (ADS)

    Bannan, Thomas; Booth, A. Murray; Alfarra, Rami; Bacak, Asan; Pericval, Carl

    2016-04-01

    Lithium ion attachment mass spectrometry provides a non-specific, non-fragmenting and sensitive method for detection of volatile species in the gas phase. The design, manufacture, and results from lithium ion attachment ionisation sources for two mass spectrometry systems are presented. Trace gas analysis is investigated using a modified Chemical Ionization Mass Spectrometer (CIMS) and vapour pressure (VP) measurements using a modified Knudsen Effusion Mass Spectrometer (KEMS) are presented. The Li+ modified CIMS provided limits of detection of 4 ppt for acetone, 0.2 ppt for formic acid, 15 ppt for nitric acid and 120 ppt from ammonia. Despite improvements, the problem of burnout remained persistent. The Li+ CIMS would unlikely be suitable for field or aircraft work, but could be appropriate for certain lab applications. The KEMS currently utilizes an electron impact (EI) ionisation source which provides a highly sensitive source, with the drawback of fragmentation of ionized molecules (Booth et al., 2009). Using Li+ KEMS the VP of samples can be measured without fragmentation and can therefore be used to identify VPs of individual components in mixtures. The validity of using Li+ for determining the VP of mixtures was tested by making single component VP measurements, which showed good agreement with EI measurements of Poly ethylene glycol (PEG) 3 and PEG 4, both when individually measured and when mixed. The Li+ KEMS was then used to investigate a system of atmospheric relevance, α-pinene secondary organic aerosol, generated in a reaction chamber (Alfarra et al., 2012). The VPs of the individual components from this generated sample are within the range we expect for compounds capable of partitioning between the particle and gas phase of an aerosol (0.1-10-5 Pa). Li+ source has a calculated sensitivity approximately 75 times less than that of EI, but the lack of fragmentation using the Li+ source is a significant advantage.

  16. Effects of interferons and double-stranded RNA on human prostate cancer cell apoptosis.

    PubMed

    Tan, Haiyan; Zeng, Chun; Xie, Junbo; Alghamdi, Norah J; Song, Ya; Zhang, Hongbing; Zhou, Aimin; Jin, Di

    2015-11-17

    Prostate cancer is the second most commonly diagnosed cancer among men in the United States. Prostate cancer therapy is severely hampered by lack of response and development of resistance to conventional chemotherapeutic drugs in patients. Therefore, the development and discovery of new drugs have become an urgent clinical need. Interferons (IFNs), a family of pleiotropic cytokines, exert antitumor activities due to their anti-proliferative, immunomodulatory and proapoptotic functions. Here, we report that pretreatment of prostate cancer PC-3 cells with IFNs sensitized these cells to double-stranded RNAs (dsRNAs)-induced apoptosis. The enhancement effect of IFN treatment was dependent on IFN subtypes, in particular, IFN γ. In comparison with IFN α or β, IFN γ treatment remarkably augmented apoptosis in PC-3 cells induced with polyinosinic:polycytidylic acid (poly I:C), a synthesized form of dsRNA. We demonstrated that IFN-signaling was necessary for these effects by using mutant cell lines. Transfection of 2-5A, the activator of RNase L, or silencing of dsRNA-dependent protein kinase R (PKR) by siRNA did not have any significant impact on this event, suggesting that neither RNase L nor PKR was involved in poly I:C/IFN γ-induced apoptosis in the cells. Further investigation of the apoptotic pathway revealed that Bak, a pro-apoptotic member of the Bcl-2family, was synergistically up-regulated by IFN γ and poly I:C, whereas other members of the family were not affected. Knocking down of Bak demonstrated its contribution to poly I:C/IFN γ-induced apoptosis in the cells. We believeour findings will precipitate the design of novel therapeutic strategies for prostate cancer. PMID:26452032

  17. Effects of interferons and double-stranded RNA on human prostate cancer cell apoptosis

    PubMed Central

    Tan, Haiyan; Zeng, Chun; Xie, Junbo; Alghamdi, Norah J.; Song, Ya; Zhang, Hongbing; Zhou, Aimin; Jin, Di

    2015-01-01

    Prostate cancer is the second most commonly diagnosed cancer among men in the United States. Prostate cancer therapy is severely hampered by lack of response and development of resistance to conventional chemotherapeutic drugs in patients. Therefore, the development and discovery of new drugs have become an urgent clinical need. Interferons (IFNs), a family of pleiotropic cytokines, exert antitumor activities due to their anti-proliferative, immunomodulatory and proapoptotic functions. Here, we report that pretreatment of prostate cancer PC-3 cells with IFNs sensitized these cells to double-stranded RNAs (dsRNAs)-induced apoptosis. The enhancement effect of IFN treatment was dependent on IFN subtypes, in particular, IFN γ. In comparison with IFN α or β, IFN γ treatment remarkably augmented apoptosis in PC-3 cells induced with polyinosinic:polycytidylic acid (poly I:C), a synthesized form of dsRNA. We demonstrated that IFN-signaling was necessary for these effects by using mutant cell lines. Transfection of 2–5A, the activator of RNase L, or silencing of dsRNA-dependent protein kinase R (PKR) by siRNA did not have any significant impact on this event, suggesting that neither RNase L nor PKR was involved in poly I:C/IFN γ-induced apoptosis in the cells. Further investigation of the apoptotic pathway revealed that Bak, a pro-apoptotic member of the Bcl-2family, was synergistically up-regulated by IFN γ and poly I:C, whereas other members of the family were not affected. Knocking down of Bak demonstrated its contribution to poly I:C/IFN γ-induced apoptosis in the cells. We believeour findings will precipitate the design of novel therapeutic strategies for prostate cancer. PMID:26452032

  18. Sodium interaction with ordered structures in mammalian red blood cells detected by Na-23 double quantum NMR.

    PubMed Central

    Shinar, H; Knubovets, T; Eliav, U; Navon, G

    1993-01-01

    Na-23 double and triple quantum filtered NMR spectra of intact dog and human red blood cells were measured with the pulse sequence 90 degrees-tau/2-180 degrees-tau/2-theta degrees-t1-theta degrees-t2(Acq). For theta = 90 degrees the triple quantum filtered spectra exhibited the typical multiple quantum filtered lineshape, characteristic of isotropic media, while the double quantum filtered ones presented a superposition of two signals, whose proportion depended on the creation time tau. This effect is due to the formation of both second and third rank tensors. The formation of the second rank tensor, T21 results from non-zero residual quadrupolar interaction and is related to the anisotropic motion of sodium ions. Measurements of the double quantum filtered spectra with theta = 54.7 degrees enabled the detection of the contribution of T21 exclusively. No residual quadrupolar interaction was detected for sodium in the cytoplasm, while unsealed ghosts displayed the double quantum filtered spectral pattern, similar to that of intact cells. The anisotropy of motion of the sodium at the plasma membrane of mammalian erythrocytes depended on the integrity of the cytoskeleton network. Theoretical analysis of the double quantum filtered spectra gave a value of residual quadrupolar splitting of approximately 20 Hz for intact unsealed ghosts. The data presented prove that double quantum filtering is a sensitive technique for detection of motional anisotropies in biological systems. Images FIGURE 5 PMID:8494983

  19. Pressure, stress, and strain distribution in the double-stage diamond anvil cell

    SciTech Connect

    Lobanov, Sergey S.; Prakapenka, Vitali B.; Prescher, Clemens; Konôpková, Zuzana; Liermann, Hanns-Peter; Crispin, Katherine L.; Zhang, Chi; Goncharov, Alexander F.

    2015-07-21

    Double stage diamond anvil cells (DACs) of two designs have been assembled and tested. We used a standard symmetric DAC with flat or beveled culets as a primary stage and CVD microanvils machined by a focused ion beam as a second. We evaluated pressure, stress, and strain distributions in gold and a mixture of gold and iron as well as in secondary anvils using synchrotron x-ray diffraction with a micro-focused beam. A maximum pressure of 240 GPa was reached independent of the first stage anvil culet size. We found that the stress field generated by the second stage anvils is typical of conventional DAC experiments. The maximum pressures reached are limited by strains developing in the secondary anvil and by cupping of the first stage diamond anvil in the presented experimental designs. Also, our experiments show that pressures of several megabars may be reached without sacrificing the first stage diamond anvils.

  20. Polymer photovoltaic cells with a graded active region achieved using double stamp transfer printing

    NASA Astrophysics Data System (ADS)

    Joo Cho, Yong; Yeob Lee, Jun; Forrest, Stephen R.

    2013-11-01

    We demonstrate that double stamp transfer printing of the poly(3-hexylthiophene) (P3HT):[6,6]-phenyl C61-butyric acid methyl ester (PCBM) active layer on MoO3 of an organic photovoltaic (OPV) cell enhances the charge collection efficiency at the anode and cathode contacts by creating a concentration gradient of the P3HT and PCBM across the bulk heterojunction active layer. This gradient increases the short circuit current and the power conversion efficiency of stamp-transferred P3HT:PCBM polymer OPVs by 23% compared with that of similarly structured spin-coated polymer OPVs due to the graded active layer composition, resulting in a power conversion efficiency of 3.7 ± 0.2% for an as-cast device. The stamp-transfer printing process provides a route to low cost fabrication of OPVs over large flexible substrate areas.

  1. DPDC (double-pass donor cell): A second-order monotone scheme for advection

    SciTech Connect

    Beason, C W; Margolin, L G

    1988-09-26

    We are developing a new, second-order, monotone scheme for advection. DPDC (i.e., double-pass donor cell) is based on Smolarkiewicz' simple, positive definite method. Both schemes are multipass methods in which upstream approximations to the truncation error are subtracted from the equations. We describe two significant improvements to Smolarkiewicz' method. First, we use a local gauge transformation to convert the method from being positive definite to the stronger condition of being monotone. Second, we analytically approximate the sum of the corrections of all the passes to use in a single corrective pass. This increases the accuracy of the method, but does not increase the order of accuracy. We compare DPDC with van Leer's method for advection of several different pulses in a constant velocity field. 5 refs., 4 figs.

  2. Influence of the fuel and dosage on the performance of double-compartment microbial fuel cells.

    PubMed

    Asensio, Y; Fernandez-Marchante, C M; Lobato, J; Cañizares, P; Rodrigo, M A

    2016-08-01

    This manuscript focuses on the evaluation of the use of different types and dosages of fuels in the performance of double-compartment microbial fuel cell equipped with carbon felt electrodes and cationic membrane. Five types of fuels (ethanol, glycerol, acetate, propionate and fructose) have been tested for the same organic load (5,000 mg L(-1) measured as COD) and for one of them (acetate), the range of dosages between 500 and 20,000 mg L(-1) of COD was also studied. Results demonstrate that production of electricity depends strongly on the fuel used. Carboxylic acids are much more efficient than alcohols or fructose for the same organic load and within the range 500-5,000 mg L(-1) of acetate the production of electricity increases linearly with the amount of acetate fed but over these concentrations a change in the population composition may explain a worse performance. PMID:27130968

  3. FACTORS AFFECTING PHYTOPLANKTON DISTRIBUTION IN A DOUBLE-CELL SEWAGE LAGOON (1) (2).

    PubMed

    DeNoyelles, F

    1967-12-01

    The distribution of phyioplankton in a double-cell sewage lagoon at Hallam, Nebraska, was studied in relation to physical, chemical, and biological factors during the summer and fall of 1965. Sixteen species of algae were recorded in the first and more organically rich of the two physically similar ponds, with 28 species recorded in the second pond. Population sizes were always greater in the first pond due to reduced grazing during the summer and large quantities of ammonia-nitrogen during the fall. The dominant algal species in both ponds on nearly all sampling dates was Ankistrodesmus falcatus v. acicularis. Declines in this population occurred with high organic pollution and heavy grazing. Both ponds had severe reductions in algal numbers during late October due to heavy grazing by the rotifer Brachionus. The distribution of phytoplankton in the Hallam ponds is compared to that of other sewage ponds in the United States, and the general pattern which emerged is discussed. PMID:27065027

  4. Doubling absorption in nanowire solar cells with dielectric shell optical antennas.

    PubMed

    Kim, Sun-Kyung; Zhang, Xing; Hill, David J; Song, Kyung-Deok; Park, Jin-Sung; Park, Hong-Gyu; Cahoon, James F

    2015-01-14

    Semiconductor nanowires (NWs) often exhibit efficient, broadband light absorption despite their relatively small size. This characteristic originates from the subwavelength dimensions and high refractive indices of the NWs, which cause a light-trapping optical antenna effect. As a result, NWs could enable high-efficiency but low-cost solar cells using small volumes of expensive semiconductor material. Nevertheless, the extent to which the antenna effect can be leveraged in devices will largely determine the economic viability of NW-based solar cells. Here, we demonstrate a simple, low-cost, and scalable route to dramatically enhance the optical antenna effect in NW photovoltaic devices by coating the wires with conformal dielectric shells. Scattering and absorption measurements on Si NWs coated with shells of SiN(x) or SiO(x) exhibit a broadband enhancement of light absorption by ∼ 50-200% and light scattering by ∼ 200-1000%. The increased light-matter interaction leads to a ∼ 80% increase in short-circuit current density in Si photovoltaic devices under 1 sun illumination. Optical simulations reproduce the experimental results and indicate the dielectric-shell effect to be a general phenomenon for groups IV, II-VI, and III-V semiconductor NWs in both lateral and vertical orientations, providing a simple route to approximately double the efficiency of NW-based solar cells. PMID:25546325

  5. Murine Double Minute-2 Prevents p53-Overactivation-Related Cell Death (Podoptosis) of Podocytes.

    PubMed

    Thomasova, Dana; Bruns, Hauke A; Kretschmer, Victoria; Ebrahim, Martrez; Romoli, Simone; Liapis, Helen; Kotb, Ahmed M; Endlich, Nicole; Anders, Hans-Joachim

    2015-07-01

    Murine double minute-2 (MDM2), an E3 ligase that regulates the cell cycle and inflammation, is highly expressed in podocytes. In podocyte injury, MDM2 drives podocyte loss by mitotic catastrophe, but the function of MDM2 in resting podocytes has not been explored. Here, we investigated the effects of podocyte MDM2 deletion in vitro and in vivo. In vitro, MDM2 knockdown by siRNA caused increased expression of p53 and podocyte death, which was completely rescued by coknockdown of p53. Apoptosis, pyroptosis, pyronecrosis, necroptosis, ferroptosis, and parthanatos were excluded as modes of occurrence for this p53-overactivation-related cell death (here referred to as podoptosis). Podoptosis was associated with cytoplasmic vacuolization, endoplasmic reticulum stress, and dysregulated autophagy (previously described as paraptosis). MDM2 knockdown caused podocyte loss and proteinuria in a zebrafish model, which was consistent with the phenotype of podocyte-specific MDM2-knockout mice that also showed the aforementioned ultrastructual podocyte abnormalities before and during progressive glomerulosclerosis. The phenotype of both animal models was entirely rescued by codeletion of p53. We conclude that MDM2 maintains homeostasis and long-term survival in podocytes by preventing podoptosis, a p53-regulated form of cell death with unspecific features previously classified as paraptosis. PMID:25349197

  6. Murine Double Minute-2 Prevents p53-Overactivation-Related Cell Death (Podoptosis) of Podocytes

    PubMed Central

    Bruns, Hauke A.; Kretschmer, Victoria; Ebrahim, Martrez; Romoli, Simone; Liapis, Helen; Kotb, Ahmed M.; Endlich, Nicole; Anders, Hans-Joachim

    2015-01-01

    Murine double minute-2 (MDM2), an E3 ligase that regulates the cell cycle and inflammation, is highly expressed in podocytes. In podocyte injury, MDM2 drives podocyte loss by mitotic catastrophe, but the function of MDM2 in resting podocytes has not been explored. Here, we investigated the effects of podocyte MDM2 deletion in vitro and in vivo. In vitro, MDM2 knockdown by siRNA caused increased expression of p53 and podocyte death, which was completely rescued by coknockdown of p53. Apoptosis, pyroptosis, pyronecrosis, necroptosis, ferroptosis, and parthanatos were excluded as modes of occurrence for this p53-overactivation-related cell death (here referred to as podoptosis). Podoptosis was associated with cytoplasmic vacuolization, endoplasmic reticulum stress, and dysregulated autophagy (previously described as paraptosis). MDM2 knockdown caused podocyte loss and proteinuria in a zebrafish model, which was consistent with the phenotype of podocyte-specific MDM2-knockout mice that also showed the aforementioned ultrastructual podocyte abnormalities before and during progressive glomerulosclerosis. The phenotype of both animal models was entirely rescued by codeletion of p53. We conclude that MDM2 maintains homeostasis and long-term survival in podocytes by preventing podoptosis, a p53-regulated form of cell death with unspecific features previously classified as paraptosis. PMID:25349197

  7. A highly durable fuel cell electrocatalyst based on double-polymer-coated carbon nanotubes

    PubMed Central

    Berber, Mohamed R.; Hafez, Inas H.; Fujigaya, Tsuyohiko; Nakashima, Naotoshi

    2015-01-01

    Driven by the demand for the commercialization of fuel cell (FC) technology, we describe the design and fabrication of a highly durable FC electrocatalyst based on double-polymer-coated carbon nanotubes for use in polymer electrolyte membrane fuel cells. The fabricated electrocatalyst is composed of Pt-deposited polybenzimidazole-coated carbon nanotubes, which are further coated with Nafion. By using this electrocatalyst, a high FC performance with a power density of 375 mW/cm2 (at 70 ˚C, 50% relative humidity using air (cathode)/H2(anode)) was obtained, and a remarkable durability of 500,000 accelerated potential cycles was recorded with only a 5% loss of the initial FC potential and 20% loss of the maximum power density, which were far superior properties compared to those of the membrane electrode assembly prepared using carbon black in place of the carbon nanotubes. The present study indicates that the prepared highly durable fuel cell electrocatalyst is a promising material for the next generation of PEMFCs. PMID:26594045

  8. Celiac Disease-Related Inflammation Is Marked by Reduction of Nkp44/Nkp46-Double Positive Natural Killer Cells

    PubMed Central

    Marafini, Irene; Monteleone, Ivan; Di Fusco, Davide; Sedda, Silvia; Cupi, Maria Laura; Fina, Daniele; Paoluzi, Alessandro Omero; Pallone, Francesco; Monteleone, Giovanni

    2016-01-01

    Introduction and Aim Natural killer (NK) cells are a first line of defence against viruses and down-regulation of NK cell cytotoxic receptors represents one of the strategies by which viruses escape the host’s immune system. Since onset of celiac disease (CD), a gluten-driven enteropathy, has been associated with viral infections, we examined whether CD-associated inflammation is characterized by abnormal distribution of NK cell receptors involved in recognition of viral-infected cells. Materials and Methods Intraepithelial mononuclear cells, isolated from duodenal biopsies of active and inactive CD patients and healthy controls (CTR) and jejunal specimens of obese subjects undergoing gastro-intestinal bypass, were analysed for NK cell markers by flow-cytometry. Expression of granzyme B, interleukin (IL)-22 and tumor necrosis factor (TNF)-α was as assessed in freshly isolated and toll-like receptor (TLR) ligand-stimulated cells. Results The percentages of total NK cells and NKT cells did not significantly differ between CD patients and CTR. In active CD, the fractions of NKp30+ NK cells, NKG2D+ NK cells and NKG2D+ NKT cells were significantly increased as compared to inactive CD patients and CTR. In contrast, CD-associated inflammation was marked by diminished presence of NKG2A+ NK cells and NKG2A+ NKT cells. The fractions of NK cells and NKT cells expressing either NKp44 or NKp46 did not differ between CD and controls, but in CD less NK cells and NKT cells co-expressed these receptors. NKp44/NKp46-double positive cells produced granzyme B and IL-22 but not TNF-α and responded to TLR ligands with enhanced expression of granzyme B. Conclusions These data indicate that active phase of CD associates with reduced presence of NKp44/NKp46-double positive NK cells and NKT cells in the epithelial compartment. PMID:27171408

  9. Financial Knudsen number: Breakdown of continuous price dynamics and asymmetric buy-and-sell structures confirmed by high-precision order-book information

    NASA Astrophysics Data System (ADS)

    Yura, Yoshihiro; Takayasu, Hideki; Sornette, Didier; Takayasu, Misako

    2015-10-01

    We generalize the description of the dynamics of the order book of financial markets in terms of a Brownian particle embedded in a fluid of incoming, exiting, and annihilating particles by presenting a model of the velocity on each side (buy and sell) independently. The improved model builds on the time-averaged number of particles in the inner layer and its change per unit time, where the inner layer is revealed by the correlations between price velocity and change in the number of particles (limit orders). This allows us to introduce the Knudsen number of the financial Brownian particle motion and its asymmetric version (on the buy and sell sides). Not being considered previously, the asymmetric Knudsen numbers are crucial in finance in order to detect asymmetric price changes. The Knudsen numbers allows us to characterize the conditions for the market dynamics to be correctly described by a continuous stochastic process. Not questioned until now for large liquid markets such as the USD-JPY and EUR-USD exchange rates, we show that there are regimes when the Knudsen numbers are so high that discrete particle effects dominate, such as during market stresses and crashes. We document the presence of imbalances of particles depletion rates on the buy and sell sides that are associated with high Knudsen numbers and violent directional price changes. This indicator can detect the direction of the price motion at the early stage while the usual volatility risk measure is blind to the price direction.

  10. Forming double layer-encapsulated quantum dots for bio-imaging and cell targeting

    NASA Astrophysics Data System (ADS)

    Fahmi, Mochamad Zakki; Chang, Jia-Yaw

    2013-01-01

    We report a simple and effective approach for the preparation of double layer-encapsulated quantum dots (DL-Qdots) composed of alkyl-capping ligands to interdigitate with hydrophobic, protective agents on the surface of AgInS2/ZnS quantum dots (Qdots), which allow phase transfer of hydrophobic Qdots from the organic phase into the aqueous phase. The alkyl-capping ligands consist of a hydrophobic, aliphatic chain and different functional terminal groups (e.g., carboxyl, amine, hydroxyl, and thiol groups) that can serve as reactive sites to chemically couple with other materials. The resulting DL-Qdots bearing various functional groups retain good fluorescence properties and show excellent solubility as well as stability over a range of pH in the aqueous phase. Cytotoxicity studies of DL-Qdots bearing carboxyl groups (DL-Qdots-COOH) were carried out against human cervical (HeLa) cancer cells to elicit no apparent toxicity even at high concentrations of 300 μg mL-1 and 24 h of incubation. To demonstrate their potential biomedical application, DL-Qdots-COOH were further conjugated with folate for staining in HeLa, human liver carcinoma (HepG2), and human breast (MCF-7) cancer cells. Confocal imaging characterization revealed that folate-conjugated DL-Qdots could target most specifically and effectively HeLa cells via folate receptor-mediated targeted delivery compared to HepG2 and MCF-7 cells. The generality and simplicity of this newly developed strategy can possibly be extended to a large variety of hydrophobic Qdots and nanocrystals whose surface protective agents have a long aliphatic chain.

  11. Efficient analysis of a small number of cancer cells at the single-cell level using an electroactive double-well array.

    PubMed

    Kim, Soo Hyeon; Fujii, Teruo

    2016-07-01

    Analysis of the intracellular materials of a small number of cancer cells at the single-cell level is important to improve our understanding of cellular heterogeneity in rare cells. To analyze an extremely small number of cancer cells (less than hundreds of cells), an efficient system is required in order to analyze target cells with minimal sample loss. Here, we present a novel approach utilizing an advanced electroactive double-well array (EdWA) for on-chip analysis of a small number of cancer cells at the single-cell level with minimal loss of target cells. The EdWA consisted of cell-sized trap-wells for deterministic single-cell trapping using dielectrophoresis and high aspect ratio reaction-wells for confining the cell lysates extracted by lysing trapped single cells via electroporation. We demonstrated a highly efficient single-cell arraying (a cell capture efficiency of 96 ± 3%) by trapping diluted human prostate cancer cells (PC3 cells). On-chip single-cell analysis was performed by measuring the intracellular β-galactosidase (β-gal) activity after lysing the trapped single cells inside a tightly enclosed EdWA in the presence of a fluorogenic enzyme substrate. The PC3 cells showed large cell-to-cell variations in β-gal activity although they were cultured under the same conditions in a culture dish. This simple and effective system has great potential for high throughput single-cell analysis of rare cells. PMID:27189335

  12. Double minute chromosomes in mouse methotrexate-resistant cells studied by atomic force microscopy

    SciTech Connect

    Deng Xinyu; Zhang Liangyu; Zhang Yu; Yan Yongda; Xu Zongwei; Dong Shen; Fu Songbin . E-mail: fusb@ems.hrbmu.edu.cn

    2006-08-11

    Double minute chromosomes (DMs) are acentric, autonomously replicating extra-chromosomes and frequently mediate gene amplification in tumor and drug resistant cells. Atomic force microscopy (AFM) is a powerful tool in microbiology. We used AFM to explore the ultrastructure of DMs in mouse fibroblasts 3T3R500. DMs in various phases of cell cycle were also studied in order to elucidate the mechanisms of their duplication and separation. Metaphase spread and induced premature condensed chromosomes (PCCs) were observed under the AFM. DMs were detected to be composed of two compact spheres linked by fibers. The fibers of DMs directly connected with metaphase chromosomes were observed. Many single-minutes and few DMs were detected in G1 PCCs, while more DMs were detected in S PCCs than in G1 PCCs. Besides, all of the DMs in G2 PCCs were coupled. Our present results suggested that DMs might divide into single-minutes during or before G1-phase, followed by duplication of the single-minutes in S-phase. Moreover, we introduced a new powerful tool to study DMs and got some ideal results.

  13. DETECTION OF EQUATORWARD MERIDIONAL FLOW AND EVIDENCE OF DOUBLE-CELL MERIDIONAL CIRCULATION INSIDE THE SUN

    SciTech Connect

    Zhao Junwei; Bogart, R. S.; Kosovichev, A. G.; Hartlep, Thomas; Duvall, T. L. Jr.

    2013-09-10

    Meridional flow in the solar interior plays an important role in redistributing angular momentum and transporting magnetic flux inside the Sun. Although it has long been recognized that the meridional flow is predominantly poleward at the Sun's surface and in its shallow interior, the location of the equatorward return flow and the meridional flow profile in the deeper interior remain unclear. Using the first 2 yr of continuous helioseismology observations from the Solar Dynamics Observatory/Helioseismic Magnetic Imager, we analyze travel times of acoustic waves that propagate through different depths of the solar interior carrying information about the solar interior dynamics. After removing a systematic center-to-limb effect in the helioseismic measurements and performing inversions for flow speed, we find that the poleward meridional flow of a speed of 15 m s{sup -1} extends in depth from the photosphere to about 0.91 R{sub Sun }. An equatorward flow of a speed of 10 m s{sup -1} is found between 0.82 and 0.91 R{sub Sun} in the middle of the convection zone. Our analysis also shows evidence of that the meridional flow turns poleward again below 0.82 R{sub Sun }, indicating an existence of a second meridional circulation cell below the shallower one. This double-cell meridional circulation profile with an equatorward flow shallower than previously thought suggests a rethinking of how magnetic field is generated and redistributed inside the Sun.

  14. Accumulation of Ku70 at DNA double-strand breaks in living epithelial cells

    SciTech Connect

    Koike, Manabu; Yutoku, Yasutomo; Koike, Aki

    2011-10-15

    Ku70 and Ku80 play an essential role in the DNA double-strand break (DSB) repair pathway, i.e., nonhomologous DNA-end-joining (NHEJ). No accumulation mechanisms of Ku70 at DSBs have been clarified in detail, although the accumulation mechanism of Ku70 at DSBs plays key roles in regulating the NHEJ activity. Here, we show the essential domains for the accumulation and function of Ku70 at DSBs in living lung epithelial cells. Our results showed that EGFP-Ku70 accumulation at DSBs began immediately after irradiation. Our findings demonstrate that three domains of Ku70, i.e., the {alpha}/{beta}, DNA-binding, and Ku80-binding domains, but not the SAP domain, are necessary for the accumulation at or recognition of DSBs in the early stage after irradiation. Moreover, our findings demonstrate that the leucine at amino acid 385 of Ku70 in the Ku80-binding domain, but not the three target amino acids for acetylation in the DNA-binding domain, is involved in the localization and accumulation of Ku70 at DSBs. Furthermore, accumulations of XRCC4 and XLF, but not that of Artemis, at DSBs are dependent on the presence of Ku70. These findings suggest that Artemis can work in not only the Ku-dependent repair process, but also the Ku-independent process at DSBs in living epithelial cells.

  15. Helicobacter pylori Infection Introduces DNA Double-Strand Breaks in Host Cells

    PubMed Central

    Hanada, Katsuhiro; Uchida, Tomohisa; Tsukamoto, Yoshiyuki; Watada, Masahide; Yamaguchi, Nahomi; Yamamoto, Kaoru; Shiota, Seiji; Moriyama, Masatsugu; Graham, David Y.

    2014-01-01

    Gastric cancer is an inflammation-related malignancy related to long-standing acute and chronic inflammation caused by infection with the human bacterial pathogen Helicobacter pylori. Inflammation can result in genomic instability. However, there are considerable data that H. pylori itself can also produce genomic instability both directly and through epigenetic pathways. Overall, the mechanisms of H. pylori-induced host genomic instabilities remain poorly understood. We used microarray screening of H. pylori-infected human gastric biopsy specimens to identify candidate genes involved in H. pylori-induced host genomic instabilities. We found upregulation of ATM expression in vivo in gastric mucosal cells infected with H. pylori. Using gastric cancer cell lines, we confirmed that the H. pylori-related activation of ATM was due to the accumulation of DNA double-strand breaks (DSBs). DSBs were observed following infection with both cag pathogenicity island (PAI)-positive and -negative strains, but the effect was more robust with cag PAI-positive strains. These results are consistent with the fact that infections with both cag PAI-positive and -negative strains are associated with gastric carcinogenesis, but the risk is higher in individuals infected with cag PAI-positive strains. PMID:25069978

  16. Quantification of cell fusion events human breast cancer cells and breast epithelial cells using a Cre-LoxP-based double fluorescence reporter system.

    PubMed

    Mohr, Marieke; Tosun, Songül; Arnold, Wolfgang H; Edenhofer, Frank; Zänker, Kurt S; Dittmar, Thomas

    2015-10-01

    The biological phenomenon of cell fusion plays an important role in several physiological processes, like fertilization, placentation, or wound healing/tissue regeneration, as well as pathophysiological processes, such as cancer. Despite this fact, considerably less is still known about the factors and conditions that will induce the merging of two plasma membranes. Inflammation and proliferation has been suggested as a positive trigger for cell fusion, but it remains unclear, which of the factor(s) of the inflamed microenvironment are being involved. To clarify this we developed a reliable assay to quantify the in vitro fusion frequency of cells using a fluorescence double reporter vector (pFDR) containing a LoxP-flanked HcRed/DsRed expression cassette followed by an EGFP expression cassette. Because cell fusion has been implicated in cancer progression four human breast cancer cell lines were stably transfected with a pFDR vector and were co-cultured with the stably Cre-expressing human breast epithelial cell line. Cell fusion is associated with a Cre-mediated recombination resulting in induction of EGFP expression in hybrid cells, which can be quantified by flow cytometry. By testing a panel of different cytokines, chemokines, growth factors and other compounds, including exosomes, under normoxic and hypoxic conditions our data indicate that the proinflammatory cytokine TNF-α together with hypoxia is a strong inducer of cell fusion in human MDA-MB-435 and MDA-MB-231 breast cancer cells. PMID:25900663

  17. A double-spike method for K-Ar measurement: A technique for high precision in situ dating on Mars and other planetary surfaces

    NASA Astrophysics Data System (ADS)

    Farley, K. A.; Hurowitz, J. A.; Asimow, P. D.; Jacobson, N. S.; Cartwright, J. A.

    2013-06-01

    A new method for K-Ar dating using a double isotope dilution technique is proposed and demonstrated. The method is designed to eliminate known difficulties facing in situ dating on planetary surfaces, especially instrument complexity and power availability. It may also have applicability in some terrestrial dating applications. Key to the method is the use of a solid tracer spike enriched in both 39Ar and 41K. When mixed with lithium borate flux in a Knudsen effusion cell, this tracer spike and a sample to be dated can be successfully fused and degassed of Ar at <1000 °C. The evolved 40Ar∗/39Ar ratio can be measured to high precision using noble gas mass spectrometry. After argon measurement the sample melt is heated to a slightly higher temperature (˜1030 °C) to volatilize potassium, and the evolved 39K/41K ratio measured by Knudsen effusion mass spectrometry. Combined with the known composition of the tracer spike, these two ratios define the K-Ar age using a single sample aliquot and without the need for extreme temperature or a mass determination. In principle the method can be implemented using a single mass spectrometer. Experiments indicate that quantitative extraction of argon from a basalt sample occurs at a sufficiently low temperature that potassium loss in this step is unimportant. Similarly, potassium isotope ratios measured in the Knudsen apparatus indicate good sample-spike equilibration and acceptably small isotopic fractionation. When applied to a flood basalt from the Viluy Traps, Siberia, a K-Ar age of 351 ± 19 Ma was obtained, a result within 1% of the independently known age. For practical reasons this measurement was made on two separate mass spectrometers, but a scheme for combining the measurements in a single analytical instrument is described. Because both parent and daughter are determined by isotope dilution, the precision on K-Ar ages obtained by the double isotope dilution method should routinely approach that of a pair of

  18. A Regulated Double-Negative Feedback Decodes the Temporal Gradient of Input Stimulation in a Cell Signaling Network.

    PubMed

    Park, Sang-Min; Shin, Sung-Young; Cho, Kwang-Hyun

    2016-01-01

    Revealing the hidden mechanism of how cells sense and react to environmental signals has been a central question in cell biology. We focused on the rate of increase of stimulation, or temporal gradient, known to cause different responses of cells. We have investigated all possible three-node enzymatic networks and identified a network motif that robustly generates a transient or sustained response by acute or gradual stimulation, respectively. We also found that a regulated double-negative feedback within the motif is essential for the temporal gradient-sensitive switching. Our analysis highlights the essential structure and mechanism enabling cells to properly respond to dynamic environmental changes. PMID:27584002

  19. Red blood cell membrane viscoelasticity, agglutination and zeta potential measurements with double optical tweezers

    NASA Astrophysics Data System (ADS)

    Fontes, Adriana; Fernandes, Heloise P.; Barjas-Castro, Maria L.; de Thomaz, André A.; de Ysasa Pozzo, Liliana; Barbosa, Luiz C.; Cesar, Carlos L.

    2006-02-01

    The red blood cell (RBC) viscoelastic membrane contains proteins and glycolproteins embedded in, or attached, to a fluid lipid bilayer and are negatively charged, which creates a repulsive electric (zeta) potential between the cells and prevents their aggregation in the blood stream. There are techniques, however, to decrease the zeta potential to allow cell agglutination which are the basis of most of the tests of antigen-antibody interactions in blood banks. This report shows the use of a double optical tweezers to measure RBC membrane viscosity, agglutination and zeta potential. In our technique one of the optical tweezers trap a silica bead that binds strongly to a RBC at the end of a RBCs rouleaux and, at the same time, acts as a pico-Newton force transducer, after calibration through its displacement from the equilibrium position. The other optical tweezers trap the RBC at the other end. To measure the membrane viscosity the optical force is measured as a function of the velocity between the RBCs. To measure the adhesion the tweezers are slowly displaced apart until the RBCs disagglutination happens. The RBC zeta potential is measured in two complimentary ways, by the force on the silica bead attached to a single RBC in response to an applied electric field, and the conventional way, by the measurement of terminal velocity of the RBC after released from the optical trap. These two measurements provide information about the RBC charges and, also, electrolytic solution properties. We believe this can improve the methods of diagnosis in blood banks.

  20. Theoretical description of the gaseous Knudsen layer in Couette flow based on the second-order constitutive and slip-jump models

    NASA Astrophysics Data System (ADS)

    Myong, R. S.

    2016-01-01

    The Knudsen layer, found in the region of gas flow very close (in order of a few mean free paths) to the solid surfaces, plays a critical role in accurately modeling rarefied and micro-scale gases. In various previous investigations, abnormal behaviors at high Knudsen numbers such as nonlinear velocity profile, velocity gradient singularity, and pronounced thermal effect are identified to exist in the Knudsen layer. However, some behaviors, in particular, the velocity gradient singularity near the surface and higher temperature, remain elusive in the continuum framework. In this study, based on the second-order macroscopic constitutive equation recently derived from the kinetic Boltzmann equation via the balanced closure and cumulant expansion [R. S. Myong, "On the high Mach number shock structure singularity caused by overreach of Maxwellian molecules," Phys. Fluids 26(5), 056102 (2014)], the macroscopic second-order constitutive and slip-jump models that are able to explain qualitatively all the known non-classical and non-isothermal behaviors are proposed. As a result, new analytical solutions to the Knudsen layer in Couette flow, in conjunction with the algebraic nonlinearly coupled second-order constitutive and Maxwell velocity slip and Smoluchowski temperature jump models, are derived. It was shown that the velocity gradient singularity in the Knudsen layer can be explained within the continuum framework, when the nonlinearity of the constitutive model is morphed into the determination of the velocity slip in the nonlinear slip and jump model. Also, the smaller velocity slip and shear stress are shown to be caused by the shear-thinning property of the second-order constitutive model, that is, vanishing effective viscosity at high Knudsen number.

  1. Gamma-irradiated quiescent cells repair directly induced double-strand breaks but accumulate persistent double-strand breaks during subsequent DNA replication.

    PubMed

    Minakawa, Yusuke; Atsumi, Yuko; Shinohara, Akira; Murakami, Yasufumi; Yoshioka, Ken-Ichi

    2016-07-01

    H2AX is expressed at very low levels in quiescent normal cells in vivo and in vitro. Such cells repair DNA double-strand breaks (DSBs) induced by γ-irradiation through a transient stabilization of H2AX. However, the resultant cells accumulate small numbers of irreparable (or persistent) DSBs via an unknown mechanism. We found that quiescent cells that had repaired DSBs directly induced by γ-rays were prone to accumulate DSBs during the subsequent DNA replication. Unlike directly induced DSBs, secondary DSBs were not efficiently repaired, although Rad51 and 53BP1 were recruited to these sites. H2AX was dramatically stabilized in response to DSBs directly caused by γ-rays, enabling γH2AX foci formation and DSB repair, whereas H2AX was barely stabilized in response to secondary DSBs, in which γH2AX foci were small and DSBs were not efficiently repaired. Our results show a pathway that leads to the persistent DSB formation after γ-irradiation. PMID:27251002

  2. Kinetic-theory predictions of clustering instabilities in granular flows: beyond the small-Knudsen-number regime

    SciTech Connect

    Mitrano, Peter P.; Zenk, John R.; Benyahia, Sofiane; Galvin, Janine E.; Dahl, Steven R.; Hrenya, Christine M.

    2013-12-04

    In this work we quantitatively assess, via instabilities, a Navier–Stokes-order (small- Knudsen-number) continuum model based on the kinetic theory analogy and applied to inelastic spheres in a homogeneous cooling system. Dissipative collisions are known to give rise to instabilities, namely velocity vortices and particle clusters, for sufficiently large domains. We compare predictions for the critical length scales required for particle clustering obtained from transient simulations using the continuum model with molecular dynamics (MD) simulations. The agreement between continuum simulations and MD simulations is excellent, particularly given the presence of well-developed velocity vortices at the onset of clustering. More specifically, spatial mapping of the local velocity-field Knudsen numbers (Knu) at the time of cluster detection reveals Knu » 1 due to the presence of large velocity gradients associated with vortices. Although kinetic-theory-based continuum models are based on a small- Kn (i.e. small-gradient) assumption, our findings suggest that, similar to molecular gases, Navier–Stokes-order (small-Kn) theories are surprisingly accurate outside their expected range of validity.

  3. Alterations in the thymocyte phenotype of EphB-deficient mice largely affect the double negative cell compartment

    PubMed Central

    Alfaro, David; Muñoz, Juan José; García-Ceca, Javier; Cejalvo, Teresa; Jiménez, Eva; Zapata, Agustín

    2008-01-01

    In the present study, we have analysed the phenotype of EphB2 and/or EphB3 deficient thymocytes confirming and extending previous studies on the role of this family of molecules in T-cell differentiation. In all mutant thymuses statistically significant reduced cell contents were observed. This reduction of thymic cellularity correlated with increased proportions of apoptotic cells, largely both double negative (DN; CD4− CD8−) and double positive (CD4+ CD8+) cells, and decreased proportions of DN cycling cells. Adult deficient thymuses also showed increased proportions of DN cells but not significant variations in the percentages of other thymocyte subsets. In absolute terms, the thymocyte number decreased significantly in all thymocyte compartments from the DN3 (CD44− CD25+) cell stage onward, without variations in the numbers of both DN1 (CD44+ CD25−) and DN2 (CD44+ CD25+) cells. Remarkably, all these changes also occurred from the 15-day fetal EphB2 and/or EphB3 deficient mice, suggesting that adult phenotype results from the gradual accumulations of defects appearing early in the thymus ontogeny. As a reflection of thymus condition, a reduction in the number of T lymphocytes occurred in the peripheral blood and mesenteric lymph nodes, but not in spleen, maintaining the proportions of T-cell subsets defined by CD4/CD8 marker expression, in all cases. PMID:18397270

  4. Reduction of myoblast differentiation following multiple population doublings in mouse C2 C12 cells: a model to investigate ageing?

    PubMed

    Sharples, Adam P; Al-Shanti, Nasser; Lewis, Mark P; Stewart, Claire E

    2011-12-01

    Ageing skeletal muscle displays declines in size, strength, and functional capacity. Given the acknowledged role that the systemic environment plays in reduced regeneration (Conboy et al. [2005] Nature 433: 760-764), the role of resident satellite cells (termed myoblasts upon activation) is relatively dismissed, where, multiple cellular divisions in-vivo throughout the lifespan could also impact on muscular deterioration. Using a model of multiple population doublings (MPD) in-vitro thus provided a system in which to investigate the direct impact of extensive cell duplications on muscle cell behavior. C(2) C(12) mouse skeletal myoblasts (CON) were used fresh or following 58 population doublings (MPD). As a result of multiple divisions, reduced morphological and biochemical (creatine kinase, CK) differentiation were observed. Furthermore, MPD cells had significantly increased cells in the S and decreased cells in the G1 phases of the cell cycle versus CON, following serum withdrawal. These results suggest continued cycling rather than G1 exit and thus reduced differentiation (myotube atrophy) occurs in MPD muscle cells. These changes were underpinned by significant reductions in transcript expression of: IGF-I and myogenic regulatory factors (myoD and myogenin) together with elevated IGFBP5. Signaling studies showed that decreased differentiation in MPD was associated with decreased phosphorylation of Akt, and with later increased phosphorylation of JNK1/2. Chemical inhibition of JNK1/2 (SP600125) in MPD cells increased IGF-I expression (non-significantly), however, did not enhance differentiation. This study provides a potential model and molecular mechanisms for deterioration in differentiation capacity in skeletal muscle cells as a consequence of multiple population doublings that would potentially contribute to the ageing process. PMID:21826704

  5. Absorption enhancement in thin film a-Si solar cells with double-sided SiO2 particle layers

    NASA Astrophysics Data System (ADS)

    Chen, Le; Wang, Qing-Kang; Shen, Xiang-Qian; Chen, Wen; Huang, Kun; Liu, Dai-Ming

    2015-10-01

    Light absorption enhancement is very important for improving the power conversion efficiency of a thin film a-Si solar cell. In this paper, a thin-film a-Si solar cell model with double-sided SiO2 particle layers is designed, and then the underlying mechanism of absorption enhancement is investigated by finite difference time domain (FDTD) simulation; finally the feasible experimental scheme for preparing the SiO2 particle layer is discussed. It is found that the top and bottom SiO2 particle layers play an important role in anti-reflection and light trapping, respectively. The light absorption of the cell with double-sided SiO2 layers greatly increases in a wavelength range of 300 nm-800 nm, and the ultimate efficiency increases more than 22% compared with that of the flat device. The cell model with double-sided SiO2 particle layers reported here can be used in varieties of thin film solar cells to further improve their performances. Project supported by the National High-Tech Research and Development Program of China (Grant No. 2011AA050518), the University Research Program of Guangxi Education Department, China (Grant No. LX2014288), and the Natural Science Foundation of Guangxi Zhuang Autonomous Region, China (Grant No. 2013GXNSBA019014).

  6. Genome Editing in Mouse Spermatogonial Stem Cell Lines Using TALEN and Double-Nicking CRISPR/Cas9.

    PubMed

    Sato, Takuya; Sakuma, Tetsushi; Yokonishi, Tetsuhiro; Katagiri, Kumiko; Kamimura, Satoshi; Ogonuki, Narumi; Ogura, Atsuo; Yamamoto, Takashi; Ogawa, Takehiko

    2015-07-14

    Mouse spermatogonial stem cells (SSCs) can be cultured for multiplication and maintained for long periods while preserving their spermatogenic ability. Although the cultured SSCs, named germline stem (GS) cells, are targets of genome modification, this process remains technically difficult. In the present study, we tested TALEN and double-nicking CRISPR/Cas9 on GS cells, targeting Rosa26 and Stra8 loci as representative genes dispensable and indispensable in spermatogenesis, respectively. Harvested GS cell colonies showed a high targeting efficiency with both TALEN and CRISPR/Cas9. The Rosa26-targeted GS cells differentiated into fertility-competent sperm following transplantation. On the other hand, Stra8-targeted GS cells showed defective spermatogenesis following transplantation, confirming its prime role in the initiation of meiosis. TALEN and CRISPR/Cas9, when applied in GS cells, will be valuable tools in the study of spermatogenesis and for revealing the genetic mechanism of spermatogenic failure. PMID:26095606

  7. DNA double-strand breaks alter the spatial arrangement of homologous loci in plant cells

    PubMed Central

    Hirakawa, Takeshi; Katagiri, Yohei; Ando, Tadashi; Matsunaga, Sachihiro

    2015-01-01

    Chromatin dynamics and arrangement are involved in many biological processes in nuclei of eukaryotes including plants. Plants have to respond rapidly to various environmental stimuli to achieve growth and development because they cannot move. It is assumed that the alteration of chromatin dynamics and arrangement support the response to these stimuli; however, there is little information in plants. In this study, we investigated the chromatin dynamics and arrangement with DNA damage in Arabidopsis thaliana by live-cell imaging with the lacO/LacI-EGFP system and simulation analysis. It was revealed that homologous loci kept a constant distance in nuclei of A. thaliana roots in general growth. We also found that DNA double-strand breaks (DSBs) induce the approach of the homologous loci with γ-irradiation. Furthermore, AtRAD54, which performs an important role in the homologous recombination repair pathway, was involved in the pairing of homologous loci with γ-irradiation. These results suggest that homologous loci approach each other to repair DSBs, and AtRAD54 mediates these phenomena. PMID:26046331

  8. Cell force mapping using a double-sided micropillar array based on the moiré fringe method

    NASA Astrophysics Data System (ADS)

    Zhang, F.; Anderson, S.; Zheng, X.; Roberts, E.; Qiu, Y.; Liao, R.; Zhang, X.

    2014-07-01

    The mapping of traction forces is crucial to understanding the means by which cells regulate their behavior and physiological function to adapt to and communicate with their local microenvironment. To this end, polymeric micropillar arrays have been used for measuring cell traction force. However, the small scale of the micropillar deflections induced by cell traction forces results in highly inefficient force analyses using conventional optical approaches; in many cases, cell forces may be below the limits of detection achieved using conventional microscopy. To address these limitations, the moiré phenomenon has been leveraged as a visualization tool for cell force mapping due to its inherent magnification effect and capacity for whole-field force measurements. This Letter reports an optomechanical cell force sensor, namely, a double-sided micropillar array (DMPA) made of poly(dimethylsiloxane), on which one side is employed to support cultured living cells while the opposing side serves as a reference pattern for generating moiré patterns. The distance between the two sides, which is a crucial parameter influencing moiré pattern contrast, is predetermined during fabrication using theoretical calculations based on the Talbot effect that aim to optimize contrast. Herein, double-sided micropillar arrays were validated by mapping mouse embryo fibroblast contraction forces and the resulting force maps compared to conventional microscopy image analyses as the reference standard. The DMPA-based approach precludes the requirement for aligning two independent periodic substrates, improves moiré contrast, and enables efficient moiré pattern generation. Furthermore, the double-sided structure readily allows for the integration of moiré-based cell force mapping into microfabricated cell culture environments or lab-on-a-chip devices.

  9. Without peripheral interference, thymic deletion is mediated in a cohort of double-positive cells without classical activation

    PubMed Central

    Zhan, Yifan; Purton, Jared F.; Godfrey, Dale I.; Cole, Timothy J.; Heath, William R.; Lew, Andrew M.

    2003-01-01

    Peripheral activation can cause bystander thymocyte death by eliciting a “cytokine storm.” This event complicates in vivo studies using exogenous ligand-induced models of negative selection. A stable transgenic model that selectively eliminates peripheral CD4 cells has allowed us to analyze negative selection as direct cognate events in two T cell receptor transgenic mice, OT-II and DO11. Whereas cognate peptide induced a massive deletion in double-positive (DP) cells in mice with peripheral CD4 cells, this DP deletion was modest in mice lacking peripheral CD4 cells. Using BrdUrd and annexin V staining, we found that negative selection primarily occurs in a cohort of DP cells and the absence of single-positive (SP) cells is largely caused by reduction in the cohort of DP precursors. Moreover, the fates of DP cells and SP cells after antigen exposure were vastly different. Whereas SP cells up-regulated uniformly their CD69 and CD44 levels, increased their cell size, and survived after antigen exposure, DP cells had less CD69 and CD44 up-regulation, no size change, and promptly died. Thus, negative selection represents an “abortive” activation different from activation-induced cell death of mature T cells. PMID:12538873

  10. DNA double-strand breaks induced in normal human cells during the repair of ultraviolet light damage.

    PubMed Central

    Bradley, M O; Taylor, V I

    1981-01-01

    DNA double-strand breaks (DSBs) are formed in normal human IMR-90 cells during repair incubation after 100 and 300 J.m-2 of ultraviolet light. By contrast, no DSBs are formed after exposure to ultraviolet light in human XPA cells (from a patient with xeroderma pigmentosum complementation group A), which are unable to excise pyrimidine dimers. The DSBs are not due to immediate cell death, because all the cells excluded trypan blue at the time of assay and because XPA cells, which are much more sensitive to ultraviolet light than are IMR-90 cells, did not form DSBs after exposure to ultraviolet light. The DSBs do not appear to be due to either DNA synthesis or cellular single-strand endonucleases. We suggest that repair-induced DSBs would be potent lesions that might lead to cytotoxicity, chromosome aberrations, deletion mutations, and perhaps cellular transformation. PMID:6267601

  11. Activity of neutral endopeptidase and aminopeptidase N in mouse thymic stromal cells which bind double-positive thymocytes.

    PubMed

    Small, M; Kaiser, M; Tse, W; Heimfeld, S; Blumberg, S

    1996-04-01

    The activity of two peptidases was determined in immortalized lines of thymic stromal cells. A line of total stromal cells (T-TG-St) was grown from transgenic mouse expressing temperature-sensitive SV40 T antigen under the control of the regulatory elements of the mouse major histocompatibility complex class I gene. From these cells we isolated a subset (DP-TG-St) that binds thymocytes which are mainly CD4+8+. We also assayed a clone of fetal thymic epithelial cells (BA/10) that binds CD4+8+ thymocytes. Both lines of double -positive cell-binding stroma exhibited strong activity of two peptidases, neutral endopeptidase (NEP; EC 3.4.24.11) and aminopeptidase N (APN; EC 3.4.11.2). In contrast, the activity of both enzymes was very low in the total thymic stromal line. Use of the specific inhibitors confirmed that these two enzymes were responsible for the activity observed but also suggested the presence of additional unidentified aminopeptidase(s) in the same stromal cells. The high activity of the two peptidases on stromal cells that bind thymocytes at the double-positive stage raises the possibility that they might contribute to the microenvironment of the developing thymocytes. PMID:8625997

  12. Thrombospondin-1 might be a therapeutic target to suppress RB cells by regulating the DNA double-strand breaks repair

    PubMed Central

    Zhang, Zhang; Zhang, Ping; Yang, Ying; Wu, Nandan; Xu, Lijun; Zhang, Jing; Ge, Jian; Yu, Keming; Zhuang, Jing

    2016-01-01

    Retinoblastoma (RB) arises from the retina, and its growth usually occurs under the retina and toward the vitreous. Ideal therapy should aim to inhibit the tumor and protect neural cells, increasing the patient's life span and quality of life. Previous studies have demonstrated that Thrombospondin-1 (TSP-1) is associated with neurogenesis, neovascularization and tumorigenesis. However, at present, the bioactivity of TSP-1 in retinoblastoma has not been defined. Herein, we demonstrated that TSP-1 was silenced in RB cell lines and clinical tumor samples. HDAC inhibitor, Trichostatin A (TSA), could notably transcriptionally up-regulate TSP-1 in RB cells, WERI-Rb1 cells and Y79 cells. Moreover, we found human recombinant TSP-1 (hTSP-1) could significantly inhibit the cell viability of RB cells both in vitro and in vivo. Interestingly, hTSP-1 could significantly induce the expression of γ-H2AX, a well-characterized in situ marker of DNA double-strand breaks (DSBs) in RB cells. The DNA NHEJ pathway in WERI-Rb1 cells could be significantly inhibited by hTSP-1. A mutation in Rb1 might be involved in the hTSP-1-medicated γ-H2AX increasing in WERI-Rb1 cells. Furthermore, hTSP-1 could inhibit RB cells while promoting retinal neurocyte survival in the neuronal and retinoblastoma cell co-culture system. As such, TSP-1 may become a therapeutic target for treatment of retinoblastoma. PMID:26756218

  13. A Subset of CD4/CD8 Double-Negative T Cells Expresses HIV Proteins in Patients on Antiretroviral Therapy

    PubMed Central

    DeMaster, Laura K.; Liu, Xiaohe; VanBelzen, D. Jake; Trinité, Benjamin; Zheng, Lingjie; Agosto, Luis M.; Migueles, Stephen A.; Connors, Mark; Sambucetti, Lidia; Levy, David N.; Pasternak, Alexander O.

    2015-01-01

    ABSTRACT A major goal in HIV eradication research is characterizing the reservoir cells that harbor HIV in the presence of antiretroviral therapy (ART), which reseed viremia after treatment is stopped. In general, it is assumed that the reservoir consists of CD4+ T cells that express no viral proteins. However, recent findings suggest that this may be an overly simplistic view and that the cells that contribute to the reservoir may be a diverse population that includes both CD4+ and CD4− cells. In this study, we directly infected resting CD4+ T cells and used fluorescence-activated cell sorting (FACS) and fiber-optic array scanning technology (FAST) to identify and image cells expressing HIV Gag. We found that Gag expression from integrated proviruses occurred in resting cells that lacked surface CD4, likely resulting from Nef- and Env-mediated receptor internalization. We also extended our approach to detect cells expressing HIV proteins in patients suppressed on ART. We found evidence that rare Gag+ cells persist during ART and that these cells are often negative for CD4. We propose that these double-negative α/β T cells that express HIV protein may be a component of the long-lived reservoir. IMPORTANCE A reservoir of infected cells persists in HIV-infected patients during antiretroviral therapy (ART) that leads to rebound of virus if treatment is stopped. In this study, we used flow cytometry and cell imaging to characterize protein expression in HIV-infected resting cells. HIV Gag protein can be directly detected in infected resting cells and occurs with simultaneous loss of CD4, consistent with the expression of additional viral proteins, such as Env and Nef. Gag+ CD4− cells can also be detected in suppressed patients, suggesting that a subset of infected cells express proteins during ART. Understanding the regulation of viral protein expression during ART will be key to designing effective strategies to eradicate HIV reservoirs. PMID:26537682

  14. Double bouquet cell in the human cerebral cortex and a comparison with other mammals.

    PubMed

    Yáñez, Inmaculada Ballesteros; Muñoz, Alberto; Contreras, Julio; Gonzalez, Juncal; Rodriguez-Veiga, Elisia; DeFelipe, Javier

    2005-06-13

    Double bouquet cells (DBCs) are neocortical gamma-aminobutyric acid (GABA)ergic interneurons characterized by the vertical bundling of its axon, which are generally termed "bundles" or "horse-tails." Using immunocytochemistry for the calcium binding protein calbindin, we have analyzed the morphology, density, and distribution of DBC horse-tails in different cortical areas of the human cortex (Brodmann's areas 10, 4, 3b, 22, 18, and 17). Although DBC horse-tails were very numerous and regularly distributed in all cortical areas, variations were observed both in terms of morphology and density. We distinguished two major classes of DBC horse-tails: the thicker complex type (type I) that had more axon collaterals; and the simple type (type II). The density of DBC horse-tails was significantly higher in areas 17, 18, 22, and 4 than in areas 3b and 10. Moreover, the proportion of type I and type II DBC horse-tails varied in the cortical areas studied. We also examined the distribution of DBC horse-tails in frontal, parietal, and occipital areas of different mammalian species. We found DBCs to be present in carnivores but not in rodents, lagomorphs, or artiodactyls. In carnivores, relatively few DBC horse-tails can be identified and they were generally found in the occipital cortex. Therefore, there is significant variability in the morphology and distribution of DBC horse-tails in different species and cortical areas. We conclude that, although these interneurons may be an important element in the organization of cortical microcolumns in primates, this is not the case in other mammalian species. PMID:15846784

  15. Use of double-stranded RNA interference in Drosophila cell lines to dissect signal transduction pathways

    PubMed Central

    Clemens, James C.; Worby, Carolyn A.; Simonson-Leff, Nancy; Muda, Marco; Maehama, Tomohiko; Hemmings, Brian A.; Dixon, Jack E.

    2000-01-01

    We demonstrate the efficacy of double-stranded RNA-mediated interference (RNAi) of gene expression in generating “knock-out” phenotypes for specific proteins in several Drosophila cell lines. We prove the applicability of this technique for studying signaling cascades by dissecting the well-characterized insulin signal transduction pathway. Specifically, we demonstrate that inhibiting the expression of the DSOR1 (mitogen-activated protein kinase kinase, MAPKK) prevents the activation of the downstream ERK-A (MAPK). In contrast, blocking ERK-A expression results in increased activation of DSOR1. We also show that Drosophila AKT (DAKT) activation depends on the insulin receptor substrate, CHICO (IRS1–4). Finally, we demonstrate that blocking the expression of Drosophila PTEN results in the activation of DAKT. In all cases, the interference of the biochemical cascade by RNAi is consistent with the known steps in the pathway. We extend this powerful technique to study two proteins, DSH3PX1 and Drosophila ACK (DACK). DSH3PX1 is an SH3, phox homology domain-containing protein, and DACK is homologous to the mammalian activated Cdc42 tyrosine kinase, ACK. Using RNAi, we demonstrate that DACK is upstream of DSH3PX1 phosphorylation, making DSH3PX1 an identified downstream target/substrate of ACK-like tyrosine kinases. These experiments highlight the usefulness of RNAi in dissecting complex biochemical signaling cascades and provide a highly effective method for determining the function of the identified genes arising from the Drosophila genome sequencing project. PMID:10823906

  16. Simultaneous double primary clear cell carcinomas of liver and kidney: a case report and review of literature.

    PubMed

    Zhang, Wei; Wang, Qiang; Jiang, Yan-Xia; Lu, Qing; Yu, Wen-Juan; Liu, Yan; Liu, Yu-Lin; Zhao, Hui; Zhuang, Jie; Li, Yu-Jun

    2015-01-01

    Reported herein are simultaneous double primary clear cell carcinomas arising from liver and kidney. A 63-year-old man underwent a partial wedge nephrectomy of the right kidney in a surgical resection for hepatocellular carcinoma. Coincidently, on histology the tumors in liver and kidney were consisted of clear cancer cells, which arranged in haphazardly intermixed pattern without capsules. Immunohistochemically, the clear neoplastic cells in the liver were positive for HepPar-1, GPC3, and negative for nephrogenic markers; however, renal clear neoplastic cells were positive for CD10, RCC and Pax-8, but negative to liver-derived markers. These findings led to the diagnosis of simultaneous double primary clear cell carcinomas of kidney and liver. Multiple primary carcinomas of kidney and liver, especially both are primary clear cell carcinomas, are extremely rare neoplasms, only 1 case has been reported previously until now. To our knowledge, this is a report of multiple primary carcinomas arising from the liver and kidney and reminders us of differentiation diagnosis with carcinoma metastasis. PMID:25755808

  17. Modeling of single event transients with dual double-exponential current sources: Implications for logic cell characterization

    SciTech Connect

    Black, Dolores A.; Robinson, William H.; Limbrick, Daniel B.; Black, Jeffrey D.; Wilcox, Ian Z.

    2015-08-07

    Single event effects (SEE) are a reliability concern for modern microelectronics. Bit corruptions can be caused by single event upsets (SEUs) in the storage cells or by sampling single event transients (SETs) from a logic path. An accurate prediction of soft error susceptibility from SETs requires good models to convert collected charge into compact descriptions of the current injection process. This paper describes a simple, yet effective, method to model the current waveform resulting from a charge collection event for SET circuit simulations. The model uses two double-exponential current sources in parallel, and the results illustrate why a conventional model based on one double-exponential source can be incomplete. A small set of logic cells with varying input conditions, drive strength, and output loading are simulated to extract the parameters for the dual double-exponential current sources. Furthermore, the parameters are based upon both the node capacitance and the restoring current (i.e., drive strength) of the logic cell.

  18. Modeling of single event transients with dual double-exponential current sources: Implications for logic cell characterization

    DOE PAGESBeta

    Black, Dolores A.; Robinson, William H.; Limbrick, Daniel B.; Black, Jeffrey D.; Wilcox, Ian Z.

    2015-08-07

    Single event effects (SEE) are a reliability concern for modern microelectronics. Bit corruptions can be caused by single event upsets (SEUs) in the storage cells or by sampling single event transients (SETs) from a logic path. An accurate prediction of soft error susceptibility from SETs requires good models to convert collected charge into compact descriptions of the current injection process. This paper describes a simple, yet effective, method to model the current waveform resulting from a charge collection event for SET circuit simulations. The model uses two double-exponential current sources in parallel, and the results illustrate why a conventional modelmore » based on one double-exponential source can be incomplete. A small set of logic cells with varying input conditions, drive strength, and output loading are simulated to extract the parameters for the dual double-exponential current sources. Furthermore, the parameters are based upon both the node capacitance and the restoring current (i.e., drive strength) of the logic cell.« less

  19. Modeling of single event transients with dual double-exponential current sources: Implications for logic cell characterization

    SciTech Connect

    Black, Dolores Archuleta; Robinson, William H.; Wilcox, Ian Zachary; Limbrick, Daniel B.; Black, Jeffrey D.

    2015-08-07

    Single event effects (SEE) are a reliability concern for modern microelectronics. Bit corruptions can be caused by single event upsets (SEUs) in the storage cells or by sampling single event transients (SETs) from a logic path. Likewise, an accurate prediction of soft error susceptibility from SETs requires good models to convert collected charge into compact descriptions of the current injection process. This paper describes a simple, yet effective, method to model the current waveform resulting from a charge collection event for SET circuit simulations. The model uses two double-exponential current sources in parallel, and the results illustrate why a conventional model based on one double-exponential source can be incomplete. Furthermore, a small set of logic cells with varying input conditions, drive strength, and output loading are simulated to extract the parameters for the dual double-exponential current sources. As a result, the parameters are based upon both the node capacitance and the restoring current (i.e., drive strength) of the logic cell.

  20. Modeling of single event transients with dual double-exponential current sources: Implications for logic cell characterization

    DOE PAGESBeta

    Black, Dolores Archuleta; Robinson, William H.; Wilcox, Ian Zachary; Limbrick, Daniel B.; Black, Jeffrey D.

    2015-08-07

    Single event effects (SEE) are a reliability concern for modern microelectronics. Bit corruptions can be caused by single event upsets (SEUs) in the storage cells or by sampling single event transients (SETs) from a logic path. Likewise, an accurate prediction of soft error susceptibility from SETs requires good models to convert collected charge into compact descriptions of the current injection process. This paper describes a simple, yet effective, method to model the current waveform resulting from a charge collection event for SET circuit simulations. The model uses two double-exponential current sources in parallel, and the results illustrate why a conventionalmore » model based on one double-exponential source can be incomplete. Furthermore, a small set of logic cells with varying input conditions, drive strength, and output loading are simulated to extract the parameters for the dual double-exponential current sources. As a result, the parameters are based upon both the node capacitance and the restoring current (i.e., drive strength) of the logic cell.« less

  1. Improved performance of Li-ion cells under pulsed load using double-layer capacitors in a hybrid circuit mode

    SciTech Connect

    ROTH,EMANUEL P.; NAGASUBRAMANIAN,GANESAN

    2000-02-07

    Electrical characteristics of hybrid power sources consisting of Li-ion cells and double-layer capacitors were studied at 25 C and {minus}20 C. The cells were initially evaluated for pulse performance and then measured in hybrid modes of operation where they were coupled with the high-power capacitors. Cells manufactured by Panasonic measured at 25 C delivered full capacities of 0.76 Ah for pulses up to 3A and cells from A and T delivered full capacities of 0.73 Ah for pulses up to 4A. Measured cell resistances were 0.15 ohms and 0.12 ohms, respectively. These measurements were repeated at {minus}20 C. Direct coupling of the cells and capacitors (coupled hybrid) using 10F Panasonic capacitors in a 8F series/parallel combination extended the full capacity pulse limits (3.0V threshold) to 5.6A for the Panasonic cells and to 9A for the A and T cells. A similar arrangement using 100F capacitors from Elna in a 60F combination increased the Panasonic cell limit to 10 A. Operation in an uncoupled hybrid mode using uncoupled cell/capacitor discharge allowed fill cell capacity usage at 25 C up to the capacitor discharge limit and showed a factor of 5 improvement in delivered capacity at {minus}20 C.

  2. Simian immunodeficiency virus infection in rhesus macaques induces selective tissue specific B cell defects in double positive CD21+CD27+ memory B cells

    PubMed Central

    Das, Arpita; Veazey, Ronald S.; Wang, Xiaolei; Lackner, Andrew A.; Xu, Huanbin; Pahar, Bapi

    2011-01-01

    B cell dysfunction represents a central feature in HIV infection and pathogenesis. Our recent studies have shown that peripheral and lymphoid double positive CD21+CD27+ B cells were able to become activated and proliferate at higher rates than other B cell subpopulations. Increased proliferation of tonsillar memory B cells were identified compared to other tissues examined. Here, we demonstrate the decreased proliferation of tonsillar memory (CD21+CD27+) B cells during acute SIV infection also suggests that these cells may play an important role in SIV pathogenesis. Our findings demonstrate that SIV infection may induce selective defective responses in specific tissues, by suppressing memory B cell proliferation in tissues. PMID:21622026

  3. T-cell chronic lymphocytic leukemia in a double yellow-headed Amazon parrot (Amazona ochrocephala oratrix).

    PubMed

    Osofsky, Anna; Hawkins, Michelle G; Foreman, Oded; Kent, Michael S; Vernau, William; Lowenstine, Linda J

    2011-12-01

    An adult, male double yellow-headed Amazon parrot (Amazona ochrocephala oratrix) was diagnosed with chronic lymphocytic leukemia based on results of a complete blood cell count and cytologic examination of a bone marrow aspirate. Treatment with oral chlorambucil was attempted, but no response was evident after 40 days. The bird was euthanatized, and the diagnosis of chronic lymphocytic leukemia was confirmed on gross and microscopic examination of tissues. Neoplastic lymphocytes were found in the bone marrow, liver, kidney, testes, and blood vessels. Based on CD3-positive immunocytochemical and immunohistochemical immunophenotyping, the chronic lymphocytic leukemia was determined to be of T-cell origin. PMID:22458185

  4. A double-fuzzy diagnostic methodology dedicated to online fault diagnosis of proton exchange membrane fuel cell stacks

    NASA Astrophysics Data System (ADS)

    Zheng, Zhixue; Péra, Marie-Cécile; Hissel, Daniel; Becherif, Mohamed; Agbli, Kréhi-Serge; Li, Yongdong

    2014-12-01

    To improve the performance and lifetime of the low temperature polymer electrolyte membrane fuel cell (PEMFC) stack, water management is an important issue. This paper aims at developing an online diagnostic methodology with the capability of discriminating different degrees of flooding/drying inside the fuel cell stack. Electrochemical impedance spectroscopy (EIS) is utilized as a basis tool and a double-fuzzy method consisting of fuzzy clustering and fuzzy logic is developed to mine diagnostic rules from the experimental data automatically. Through online experimental verification, a high interpretability and computational efficiency of the proposed methodology can be achieved.

  5. Simulation and optimization of current and lattice matching double-junction GaNAsP/Si solar cells

    NASA Astrophysics Data System (ADS)

    Nacer, S.; Aissat, A.

    2016-01-01

    This paper deals with theoretical investigation of the performance of current and lattice matched GaNxAsyP1-x-y/Si double-junction solar cells. The nitrogen and arsenic concentrations ensuring lattice matching to Si are determined. The band gap of GaNAsP is calculated using the band anti-crossing model. Calculations were performed under 1-sun AM1.5 using the one diode ideal model. Impact of minor carrier lifetime and surface recombination in the top sub-cell on the cell performances is analyzed. Optimum compositions of the top sub-cell have been identified (x = 4.5%, y = 11.5% and Eg = 1.68 eV). The simulation results predict, for the optimized GaNAsP/Si double-junction solar cell, a short circuit current Jsc = 20 mA/cm2, an open circuit voltage Voc = 1.95 V, and a conversion efficiency η = 37.5%.

  6. Interleukin-2 Enhances the Regulatory Functions of CD4(+)T Cell-Derived CD4(-)CD8(-) Double Negative T Cells.

    PubMed

    Cong, Min; Liu, Tianhui; Tian, Dan; Guo, Hongbo; Wang, Ping; Liu, Kai; Lin, Jun; Tian, Yue; Shi, Wen; You, Hong; Jia, Jidong; Zhang, Dong

    2016-08-01

    CD4(+) T cells can be converted to CD4(-)CD8(-) double negative T cells (DN T cells) under appropriate conditions, and IL-2 enhanced the conversion. Here, we investigated the effect of IL-2 on the proliferation and function of converted DN T cells in vitro and in vivo. DN T cells were hyporesponsive when restimulated by mature dendritic cells (mDCs), IL-2 completely restored their responsiveness in vitro. In addition, IL-2 increased the resistance of DN T cells to apoptosis in vivo. DN T cells profoundly inhibited the proliferation of CD4(+)CD25(-) T effector cells triggered by mDCs in vitro, and this suppression was further enhanced by IL-2. Adoptively transferring of DN T cells, in combination with IL-2, inhibited the proliferation and enhanced apoptosis of alloreactive CD4(+) T cells, which resulted in significant prolongation of skin allograft survival time. Perforin plays a key role in the enhancement of DN T cells immune regulation by IL-2. In conclusion, we elucidated that IL-2 promoted DN T cell proliferation and suppressive function. The combination of DN T cells and exogenous IL-2 may represent a novel therapy in the clinical setting to prevent allograft rejection and induce immune tolerance. PMID:27135902

  7. Double Positive CD4CD8 αβ T Cells: A New Tumor-Reactive Population in Human Melanomas

    PubMed Central

    Desfrançois, Juliette; Moreau-Aubry, Agnès; Vignard, Virginie; Godet, Yann; Khammari, Amir; Dréno, Brigitte; Jotereau, Francine; Gervois, Nadine

    2010-01-01

    Background Double positive (DP) CD4CD8 Tαβ cells have been reported in normal individuals as well as in different pathological conditions including inflammatory diseases, viral infections and cancer, but their function remains to be elucidated. We recently reported the increased frequency of DP Tαβ cells in human breast pleural effusions. This manuscript addresses the question of the existence and above all the role of this non-conventional DP sub-population among tumor associated lymphocytes in melanomas. Methodology/Principal Findings We analyzed the intratumoral cell infiltrate in solid metastasis (n = 6) and tumor invaded lymph nodes (n = 26) samples from melanomas patients by multiparametric cytometry. Here we documented for the first time significant increased frequency of DP T cells in about 60% of melanoma tumors compared to blood samples. Interestingly, a high proportion of these cells produced TNF-α in response to autologous melanoma cell lines. Besides, they are characterized by a unique cytokine profile corresponding to higher secretion of IL-13, IL-4 and IL-5 than simple positive T cells. In deep analysis, we derived a representative tumor-reactive DP T cell clone from a melanoma patient's invaded lymph node. This clone was restricted by HLA-A*2402 and recognized both autologous and allogeneic tumor cells of various origins as well as normal cells, suggesting that the target antigen was a ubiquitous self antigen. However, this DP T cell clone failed to kill HLA-A*2402 EBV-transformed B cells, probably due to the constitutive expression of immunoproteasome by these cells. Conclusions/Significance In conclusion, we can postulate that, according to their broad tumor reactivity and to their original cytokine profile, the tumor associated DP T cells could participate in immune responses to tumors in vivo. Therefore, the presence of these cells and their role will be crucial to address in cancer patients, especially in the context of

  8. Double-Positive CD21+CD27+ B Cells Are Highly Proliferating Memory Cells and Their Distribution Differs in Mucosal and Peripheral Tissues

    PubMed Central

    Das, Arpita; Xu, Huanbin; Wang, Xiaolei; Yau, Canddy L.; Veazey, Ronald S.; Pahar, Bapi

    2011-01-01

    Background Several B-cell defects arise in HIV infected patients, particularly in patients with chronic infection and high viral load. Loss of memory B cells (CD27+ B cells) in peripheral blood and lymphoid tissues is one of the major B cell dysfunctions in HIV and simian immunodeficiency virus (SIV) infection. Despite several studies, definitive identification of memory B cells based on CD27 surface expression has not been described. Similarly, the rates of cell turnover in different B cell subpopulation from lymphoid and mucosal tissues have not been well documented. In this study, we demonstrate the presence of memory B cell populations and define their distribution, frequency and immunophenotype with regards to activation, proliferation, maturation, and antibody production in normal rhesus macaques from different lymphoid tissues. Methodology/Principal Findings Thirteen healthy, uninfected rhesus macaques were selected for this study. CD20+ B cells were isolated from peripheral blood and sorted based on CD27 and CD21 surface markers to define memory B cell population. All the B cell subpopulation was further characterized phenotypically and their cell turnover rates were evaluated in vivo following bromodeoxyuridine (BrdU) inoculation. Double positive (DP) CD21+CD27+ B cells in both peripheral and lymphoid tissues are memory B cells, able to produce antibody by polyclonal activation, and without T cell help. Peripheral and lymphoid DP CD21+CD27+ B cells were also able to become activated and proliferate at higher rates than other B cell subpopulations. Increased turnover of tonsillar memory B cells were identified compared to other tissues examined. Conclusions/Significance We suggest that this DP memory B cells play a major role in the immune system and their function and proliferation might have an important role in HIV/SIV mediated B cell dysregulation and pathogenesis. PMID:21304587

  9. Double stranded RNA-dependent protein kinase is involved in osteoclast differentiation of RAW264.7 cells in vitro

    SciTech Connect

    Teramachi, Junpei; Morimoto, Hiroyuki; Baba, Ryoko; Doi, Yoshiaki; Hirashima, Kanji; Haneji, Tatsuji

    2010-11-15

    Double-stranded RNA-dependent protein kinase (PKR) plays a critical role in antiviral defence of the host cells. PKR is also involved in cell cycle progression, cell proliferation, cell differentiation, tumorigenesis, and apoptosis. We previously reported that PKR is required for differentiation and calcification of osteoblasts. However, it is unknown about the role of PKR in osteoclast differentiation. A dominant-negative PKR mutant cDNA, in which the amino acid lysine at 296 was replaced with arginine, was transfected into RAW264.7 cells. We have established the cell line that stably expresses the PKR mutant gene (PKR-K/R). Phosphorylation of PKR and {alpha}-subunit of eukaryotic initiation factor 2 was not stimulated by polyinosic-polycytidylic acid in the PKR-K/R cells. RANKL stimulated the formation of TRAP-positive multinuclear cells in RAW264.7 cells. However, TRAP-positive multinuclear cells were not formed in the PKR-K/R cells even when the cells were stimulated with higher doses of RANKL. A specific inhibitor of PKR, 2-aminopurine, also suppressed the RANKL-induced osteoclast differentiation in RAW264.7 cells. The expression of macrophage fusion receptor and dendritic cell-specific transmembrane protein significantly decreased in the PKR-K/R cells by real time PCR analysis. The results of RT-PCR revealed that the mRNA expression of osteoclast markers (cathepsin K and calcitonin receptor) was suppressed in the PKR-K/R cells and RAW264.7 cells treated with 2-aminopurine. Expression of NF-{kappa}B protein was suppressed in the PKR-K/R cells and 2-aminopurine-treated RAW264.7 cells. The level of STAT1 protein expression was elevated in the PKR-K/R cells compared with that of the wild-type cells. Immunohistochemical study showed that PKR was localized in osteoclasts of metatarsal bone of newborn mouse. The finding that the PKR-positive multinuclear cells should be osteoclasts was confirmed by TRAP-staining. Our present study indicates that PKR plays important

  10. Oncogenic ras-driven cancer cell vesiculation leads to emission of double-stranded DNA capable of interacting with target cells

    SciTech Connect

    Lee, Tae Hoon; Chennakrishnaiah, Shilpa; Audemard, Eric; Montermini, Laura; Meehan, Brian; Rak, Janusz

    2014-08-22

    Highlights: • Oncogenic H-ras stimulates emission of extracellular vesicles containing double-stranded DNA. • Vesicle-associated extracellular DNA contains mutant N-ras sequences. • Vesicles mediate intercellular transfer of mutant H-ras DNA to normal fibroblasts where it remains for several weeks. • Fibroblasts exposed to vesicles containing H-ras DNA exhibit increased proliferation. - Abstract: Cell free DNA is often regarded as a source of genetic cancer biomarkers, but the related mechanisms of DNA release, composition and biological activity remain unclear. Here we show that rat epithelial cell transformation by the human H-ras oncogene leads to an increase in production of small, exosomal-like extracellular vesicles by viable cancer cells. These EVs contain chromatin-associated double-stranded DNA fragments covering the entire host genome, including full-length H-ras. Oncogenic N-ras and SV40LT sequences were also found in EVs emitted from spontaneous mouse brain tumor cells. Disruption of acidic sphingomyelinase and the p53/Rb pathway did not block emission of EV-related oncogenic DNA. Exposure of non-transformed RAT-1 cells to EVs containing mutant H-ras DNA led to the uptake and retention of this material for an extended (30 days) but transient period of time, and stimulated cell proliferation. Thus, our study suggests that H-ras-mediated transformation stimulates vesicular emission of this histone-bound oncogene, which may interact with non-transformed cells.

  11. Biosynthesis of major histocompatibility complex molecules and generation of T cells in Ii TAP1 double-mutant mice.

    PubMed Central

    Tourne, S; van Santen, H M; van Roon, M; Berns, A; Benoist, C; Mathis, D; Ploegh, H

    1996-01-01

    Major histocompatibility complex (MHC) class I and II molecules are loaded with peptides in distinct subcellular compartments. The transporter associated with antigen processing (TAP) is responsible for delivering peptides derived from cytosolic proteins to the endoplasmic reticulum, where they bind to class I molecules, while the invariant chain (Ii) directs class II molecules to endosomal compartments, where they bind peptides originating mostly from exogenous sources. Mice carrying null mutations of the TAP1 or Ii genes (TAP10) or Ii0, respectively) have been useful tools for elucidating the two MHC/peptide loading pathways. To evaluate to what extent these pathways functionally intersect, we have studied the biosynthesis of MHC molecules and the generation of T cells in Ii0TAP10 double-mutant mice. We find that the assembly and expression of class II molecules in Ii0 and Ii0TAP10 animals are indistinguishable and that formation and display of class I molecules is the same in TAP10 and Ii0TAP10 animals. Thymic selection in the double mutants is as expected, with reduced numbers of both CD4+ CD8- and CD4- CD8+ thymocyte compartments. Surprisingly, lymph node T-cell populations look almost normal; we propose that population expansion of peripheral T cells normalizes the numbers of CD4+ and CD8+ cells in Ii0TAP10 mice. Images Fig. 1 Fig. 2 PMID:8643655

  12. Activation of double-stranded RNA-dependent protein kinase inhibits proliferation of pancreatic β-cells

    SciTech Connect

    Chen, Shan-Shan; Jiang, Teng; Wang, Yi; Gu, Li-Ze; Wu, Hui-Wen; Tan, Lan; Guo, Jun

    2014-01-17

    Highlights: •PKR can be activated by glucolipitoxicity and pro-inflammatory cytokines in β-cells. •Activated PKR inhibited β-cell proliferation by arresting cell cycle at G1 phase. •Activated PKR fully abrogated the pro-proliferative effects of IGF-I on β-cells. -- Abstract: Double-stranded RNA-dependent protein kinase (PKR) is revealed to participate in the development of insulin resistance in peripheral tissues in type 2 diabetes (T2DM). Meanwhile, PKR is also characterized as a critical regulator of cell proliferation. To date, no study has focused on the impact of PKR on the proliferation of pancreatic β-cells. Here, we adopted insulinoma cell lines and mice islet β-cells to investigate: (1) the effects of glucolipotoxicity and pro-inflammatory cytokines on PKR activation; (2) the effects of PKR on proliferation of pancreatic β-cells and its underlying mechanisms; (3) the actions of PKR on pro-proliferative effects of IGF-I and its underlying pathway. Our results provided the first evidence that PKR can be activated by glucolipitoxicity and pro-inflammatory cytokines in pancreatic β-cells, and activated PKR significantly inhibited cell proliferation by arresting cell cycle at G1 phase. Reductions in cyclin D1 and D2 as well as increases in p27 and p53 were associated with the anti-proliferative effects of PKR, and proteasome-dependent degradation took part in the reduction of cyclin D1 and D2. Besides, PKR activation abrogated the pro-proliferative effects of IGF-I by activating JNK and disrupting IRS1/PI3K/Akt signaling pathway. These findings indicate that the anti-proliferative actions of PKR on pancreatic β-cells may contribute to the pathogenesis of T2DM.

  13. Uracil DNA glycosylase (UNG) loss enhances DNA double strand break formation in human cancer cells exposed to pemetrexed

    PubMed Central

    Weeks, L D; Zentner, G E; Scacheri, P C; Gerson, S L

    2014-01-01

    Misincorporation of genomic uracil and formation of DNA double strand breaks (DSBs) are known consequences of exposure to TS inhibitors such as pemetrexed. Uracil DNA glycosylase (UNG) catalyzes the excision of uracil from DNA and initiates DNA base excision repair (BER). To better define the relationship between UNG activity and pemetrexed anticancer activity, we have investigated DNA damage, DSB formation, DSB repair capacity, and replication fork stability in UNG+/+ and UNG−/− cells. We report that despite identical growth rates and DSB repair capacities, UNG−/− cells accumulated significantly greater uracil and DSBs compared with UNG+/+ cells when exposed to pemetrexed. ChIP-seq analysis of γ-H2AX enrichment confirmed fewer DSBs in UNG+/+ cells. Furthermore, DSBs in UNG+/+ and UNG−/− cells occur at distinct genomic loci, supporting differential mechanisms of DSB formation in UNG-competent and UNG-deficient cells. UNG−/− cells also showed increased evidence of replication fork instability (PCNA dispersal) when exposed to pemetrexed. Thymidine co-treatment rescues S-phase arrest in both UNG+/+ and UNG−/− cells treated with IC50-level pemetrexed. However, following pemetrexed exposure, UNG−/− but not UNG+/+ cells are refractory to thymidine rescue, suggesting that deficient uracil excision rather than dTTP depletion is the barrier to cell cycle progression in UNG−/− cells. Based on these findings we propose that pemetrexed-induced uracil misincorporation is genotoxic, contributing to replication fork instability, DSB formation and ultimately cell death. PMID:24503537

  14. A new technique for extracting physical parameters of a solar cell model from the double exponential model (DECM)

    NASA Astrophysics Data System (ADS)

    Yadir, S.; Assal, S.; El Rhassouli, A.; Sidki, M.; Benhmida, M.

    2013-11-01

    In this paper, we propose and apply a new technique for extracting physical parameters of solar cell double exponential model with two ideality factor constants (DECM) from illuminated current-voltage (I-V) experimental characteristics. The equivalent circuit of solar cell includes two constant diodes ideality factors (n1 = 1, n2 = 2) with two saturation currents I0D and I0R, a current generator intensity Iph, a series resistor RS and a conductance GP. A set of current-voltage characteristics are generated by injecting various RS values in the characteristic equation. Using the area error rate ("%ΔArea,") between the experimental and extracted (I-V) characteristics, the value of RS is deduced as the minimum of this error. The obtained results show a good agreement with the experimental characteristics measured on a commercial polycrystalline solar cell.

  15. Mixed metal oxides for dye-sensitized solar cell using zinc titanium layered double hydroxide as precursor

    NASA Astrophysics Data System (ADS)

    Liu, Jianqiang; Qin, Yaowei; Zhang, Liangji; Xiao, Hongdi; Song, Jianye; Liu, Dehe; Leng, Mingzhe; Hou, Wanguo; Du, Na

    2013-12-01

    Mixed metal oxides (MMO) are always obtained from layered double hydroxide (LDH) by thermal decomposition. In the present work, a zinc titanium LDH with the zinc titanium molar ratio of 4.25 was prepared by urea method and ZnO-based mixed oxides were obtained by calcining at or over 500°C. The MMO was used as electrodes for dye sensitized solar cell (DSSC). The cells constructed by films of prepared composite materials using a N719 as dye were prepared. The efficiency values of these cells are 0.691%, 0.572% and 0.302% with MMO prepared at 500, 600 and 700°C, respectively.

  16. Application of laser-accelerated protons to the demonstration of DNA double-strand breaks in human cancer cells

    NASA Astrophysics Data System (ADS)

    Yogo, A.; Sato, K.; Nishikino, M.; Mori, M.; Teshima, T.; Numasaki, H.; Murakami, M.; Demizu, Y.; Akagi, S.; Nagayama, S.; Ogura, K.; Sagisaka, A.; Orimo, S.; Nishiuchi, M.; Pirozhkov, A. S.; Ikegami, M.; Tampo, M.; Sakaki, H.; Suzuki, M.; Daito, I.; Oishi, Y.; Sugiyama, H.; Kiriyama, H.; Okada, H.; Kanazawa, S.; Kondo, S.; Shimomura, T.; Nakai, Y.; Tanoue, M.; Sasao, H.; Wakai, D.; Bolton, P. R.; Daido, H.

    2009-05-01

    We report the demonstrated irradiation effect of laser-accelerated protons on human cancer cells. In vitro (living) A549 cells are irradiated with quasimonoenergetic proton bunches of 0.8-2.4 MeV with a single bunch duration of 15 ns. Irradiation with the proton dose of 20 Gy results in a distinct formation of γ-H2AX foci as an indicator of DNA double-strand breaks generated in the cancer cells. This is a pioneering result that points to future investigations of the radiobiological effects of laser-driven ion beams. Unique high-current and short-bunch features make laser-driven proton bunches an excitation source for time-resolved determination of radical yields.

  17. Application of laser-accelerated protons to the demonstration of DNA double-strand breaks in human cancer cells

    SciTech Connect

    Yogo, A.; Nishikino, M.; Mori, M.; Ogura, K.; Sagisaka, A.; Orimo, S.; Nishiuchi, M.; Pirozhkov, A. S.; Ikegami, M.; Tampo, M.; Sakaki, H.; Suzuki, M.; Daito, I.; Kiriyama, H.; Okada, H.; Kanazawa, S.; Kondo, S.; Shimomura, T.; Nakai, Y.; Bolton, P. R.

    2009-05-04

    We report the demonstrated irradiation effect of laser-accelerated protons on human cancer cells. In vitro (living) A549 cells are irradiated with quasimonoenergetic proton bunches of 0.8-2.4 MeV with a single bunch duration of 15 ns. Irradiation with the proton dose of 20 Gy results in a distinct formation of {gamma}-H2AX foci as an indicator of DNA double-strand breaks generated in the cancer cells. This is a pioneering result that points to future investigations of the radiobiological effects of laser-driven ion beams. Unique high-current and short-bunch features make laser-driven proton bunches an excitation source for time-resolved determination of radical yields.

  18. A double transgenic mouse used to track migrating Schwann cells and regenerating axons following engraftment of injured nerves

    PubMed Central

    Hayashi, Ayato; Koob, Jason W; Liu, Daniel Z; Tong, Alice Y; Hunter, Daniel A.; Parsadanian, Alexander; Mackinnon, Susan E.; Myckatyn, Terence M.

    2007-01-01

    We propose that double transgenic thy1-CFP(23)/S100-GFP mice whose Schwann cells constitutively express green fluorescent protein (GFP) and axons express cyan fluorescent protein (CFP) can be used to serially evaluate the temporal relationship between nerve regeneration and Schwann cell migration through acellular nerve grafts. Thy1-CFP(23)/S100-GFP and S100-GFP mice received non-fluorescing cold preserved nerve allografts from immunologically disparate donors. In vivo fluorescent imaging of these grafts was then performed at multiple points. The transected sciatic nerve was reconstructed with a 1 cm nerve allograft harvested from a Balb-C mouse and acellularized via 7 weeks of cold preservation prior to transplantation. The presence of regenerated axons and migrating Schwann cells was confirmed with confocal and electron microscopy on fixed tissue. Schwann cells migrated into the acellular graft (163 ± 15 intensity units) from both proximal and distal stumps, and bridged the whole graft within 10 days (388 ± 107 intensity units in the central 4-6 mm segment). Nerve regeneration lagged behind Schwann cell migration with 5 or 6 axons imaged traversing the proximal 4 mm of the graft under confocal microcopy within 10 days, and up to 21 labeled axons crossing the distal coaptation site by 15 days. Corroborative electron and light microscopy 5 mm into the graft demonstrated relatively narrow diameter myelinated (431±31) and unmyelinated (64±9) axons by 28 but not 10 days. Live imaging of the double-transgenic thy1-CFP(23)/S100-GFP murine line enabled serial assessment of Schwann cell-axonal relationships in traumatic nerve injuries reconstructed with acellular nerve allografts. PMID:17628544

  19. Myricetin induces apoptosis via endoplasmic reticulum stress and DNA double-strand breaks in human ovarian cancer cells

    PubMed Central

    XU, YE; XIE, QI; WU, SHAOHUA; YI, DAN; YU, YANG; LIU, SHIBING; LI, SONGYAN; LI, ZHIXIN

    2016-01-01

    The mechanisms underlying myricetin-induced cancer cell apoptosis remain to be elucidated. Certain previous studies have shown that myricetin induces apoptosis through the mitochondrial pathway. Apoptosis, however, can also be induced by other classical pathways, including endoplasmic reticulum (ER) stress and DNA double-strand breaks (DSBs). The aim of the present study was to assess whether these two apoptotic pathways are involved in myricetin-induced cell death in SKOV3 ovarian cancer cells. The results revealed that treatment with myricetin inhibited viability of SKOV3 cells in a dose-dependent manner. Myricetin induced nuclear chromatin condensation and fragmentation, and also upregulated the protein levels of active caspase 3 in a time-dependent manner. In addition, myricetin upregulated ER stress-associated proteins, glucose-regulated protein-78 and C/EBP homologous protein in SKOV3 cells. Phosphorylation of H2AX, a marker of DNA DSBs, was revealed to be upregulated in myricetin-treated cells. The data indicated that myricetin induces DNA DSBs and ER stress, which leads to apoptosis in SKOV3 cells. PMID:26782830

  20. Theoretica Study of Asymmetric Double D-π-A Organic Sensitizers for Efficient Dye-Sensitized Solar Cells.

    PubMed

    Kwon, Dong Yuel; Lee, Gun Hyung; Kim, Young Sik

    2015-03-01

    Three novel dye sensitizers that were based on asymmetric double D-π-A chains with phenoxazine (POZ) and diphenylamine (DPA) as electron donors and cyanoacetic acid (CA) and 2-(1,1- dicyanomethylene) rhodanine (RD) as electron acceptors (DCPR, DRPC, DRPR) were designed, theoretically investigated, and compared with the reference dye based on asymmetric double D-π-A chains (DCPC). Using density functional theory (DFT) and time-dependent density functional theory (TD-DFT) calculations, we gained insight into the factors responsible for the photovoltaic properties of the dye sensitizers. Due to the different HOMO levels of each donor and the different LUMO levels of each acceptor, the absorption spectrum of each dye showed different shapes. Among the dyes, DRPR showed a broader and more bathochromically shifted absorption band than the other dies. It also showed a higher molar extinction coefficient than that of the reference dye (DCPC). This work suggests optimizing the chain of electron donors and acceptors in dye sensitizers based on asymmetric double D-π-A chains would produce good photovoltaic properties for dye-sensitized solar cells (DSSCs). PMID:26413690

  1. Novel Double-Hit Model of Radiation and Hyperoxia-Induced Oxidative Cell Damage Relevant to Space Travel.

    PubMed

    Pietrofesa, Ralph A; Velalopoulou, Anastasia; Lehman, Stacey L; Arguiri, Evguenia; Solomides, Pantelis; Koch, Cameron J; Mishra, Om P; Koumenis, Constantinos; Goodwin, Thomas J; Christofidou-Solomidou, Melpo

    2016-01-01

    Spaceflight occasionally requires multiple extravehicular activities (EVA) that potentially subject astronauts to repeated changes in ambient oxygen superimposed on those of space radiation exposure. We thus developed a novel in vitro model system to test lung cell damage following repeated exposure to radiation and hyperoxia. Non-tumorigenic murine alveolar type II epithelial cells (C10) were exposed to >95% O₂ for 8 h only (O₂), 0.25 Gy ionizing γ-radiation (IR) only, or a double-hit combination of both challenges (O₂ + IR) followed by 16 h of normoxia (ambient air containing 21% O₂ and 5% CO₂) (1 cycle = 24 h, 2 cycles = 48 h). Cell survival, DNA damage, apoptosis, and indicators of oxidative stress were evaluated after 1 and 2 cycles of exposure. We observed a significant (p < 0.05) decrease in cell survival across all challenge conditions along with an increase in DNA damage, determined by Comet analysis and H2AX phosphorylation, and apoptosis, determined by Annexin-V staining, relative to cells unexposed to hyperoxia or radiation. DNA damage (GADD45α and cleaved-PARP), apoptotic (cleaved caspase-3 and BAX), and antioxidant (HO-1 and Nqo1) proteins were increased following radiation and hyperoxia exposure after 1 and 2 cycles of exposure. Importantly, exposure to combination challenge O₂ + IR exacerbated cell death and DNA damage compared to individual exposures O₂ or IR alone. Additionally levels of cell cycle proteins phospho-p53 and p21 were significantly increased, while levels of CDK1 and Cyclin B1 were decreased at both time points for all exposure groups. Similarly, proteins involved in cell cycle arrest was more profoundly changed with the combination challenges as compared to each stressor alone. These results correlate with a significant 4- to 6-fold increase in the ratio of cells in G2/G1 after 2 cycles of exposure to hyperoxic conditions. We have characterized a novel in vitro model of double-hit, low-level radiation and hyperoxia

  2. Novel Double-Hit Model of Radiation and Hyperoxia-Induced Oxidative Cell Damage Relevant to Space Travel

    PubMed Central

    Pietrofesa, Ralph A.; Velalopoulou, Anastasia; Lehman, Stacey L.; Arguiri, Evguenia; Solomides, Pantelis; Koch, Cameron J.; Mishra, Om P.; Koumenis, Constantinos; Goodwin, Thomas J.; Christofidou-Solomidou, Melpo

    2016-01-01

    Spaceflight occasionally requires multiple extravehicular activities (EVA) that potentially subject astronauts to repeated changes in ambient oxygen superimposed on those of space radiation exposure. We thus developed a novel in vitro model system to test lung cell damage following repeated exposure to radiation and hyperoxia. Non-tumorigenic murine alveolar type II epithelial cells (C10) were exposed to >95% O2 for 8 h only (O2), 0.25 Gy ionizing γ-radiation (IR) only, or a double-hit combination of both challenges (O2 + IR) followed by 16 h of normoxia (ambient air containing 21% O2 and 5% CO2) (1 cycle = 24 h, 2 cycles = 48 h). Cell survival, DNA damage, apoptosis, and indicators of oxidative stress were evaluated after 1 and 2 cycles of exposure. We observed a significant (p < 0.05) decrease in cell survival across all challenge conditions along with an increase in DNA damage, determined by Comet analysis and H2AX phosphorylation, and apoptosis, determined by Annexin-V staining, relative to cells unexposed to hyperoxia or radiation. DNA damage (GADD45α and cleaved-PARP), apoptotic (cleaved caspase-3 and BAX), and antioxidant (HO-1 and Nqo1) proteins were increased following radiation and hyperoxia exposure after 1 and 2 cycles of exposure. Importantly, exposure to combination challenge O2 + IR exacerbated cell death and DNA damage compared to individual exposures O2 or IR alone. Additionally levels of cell cycle proteins phospho-p53 and p21 were significantly increased, while levels of CDK1 and Cyclin B1 were decreased at both time points for all exposure groups. Similarly, proteins involved in cell cycle arrest was more profoundly changed with the combination challenges as compared to each stressor alone. These results correlate with a significant 4- to 6-fold increase in the ratio of cells in G2/G1 after 2 cycles of exposure to hyperoxic conditions. We have characterized a novel in vitro model of double-hit, low-level radiation and hyperoxia exposure that

  3. Biomimetic collagen I and IV double layer Langmuir-Schaefer films as microenvironment for human pluripotent stem cell derived retinal pigment epithelial cells.

    PubMed

    Sorkio, Anni E; Vuorimaa-Laukkanen, Elina P; Hakola, Hanna M; Liang, Huamin; Ujula, Tiina A; Valle-Delgado, Juan José; Österberg, Monika; Yliperttula, Marjo L; Skottman, Heli

    2015-05-01

    The environmental cues received by the cells from synthetic substrates in vitro are very different from those they receive in vivo. In this study, we applied the Langmuir-Schaefer (LS) deposition, a variant of Langmuir-Blodgett technique, to fabricate a biomimetic microenvironment mimicking the structure and organization of native Bruch's membrane for the production of the functional human embryonic stem cell derived retinal pigment epithelial (hESC-RPE) cells. Surface pressure-area isotherms were measured simultaneously with Brewster angle microscopy to investigate the self-assembly of human collagens type I and IV on air-subphase interface. Furthermore, the structure of the prepared collagen LS films was characterized with scanning electron microscopy, atomic force microscopy, surface plasmon resonance measurements and immunofluorescent staining. The integrity of hESC-RPE on double layer LS films was investigated by measuring transepithelial resistance and permeability of small molecular weight substance. Maturation and functionality of hESC-RPE cells on double layer collagen LS films was further assessed by RPE-specific gene and protein expression, growth factor secretion, and phagocytic activity. Here, we demonstrated that the prepared collagen LS films have layered structure with oriented fibers corresponding to architecture of the uppermost layers of Bruch's membrane and result in increased barrier properties and functionality of hESC-RPE cells as compared to the commonly used dip-coated controls. PMID:25771016

  4. Catalyzed double layer cathodes for high performance and long life molten carbonate fuel cells

    SciTech Connect

    Bischoff, M.; Jantsch, U.; Rohland, B.

    1996-12-31

    NiO/LiCoO{sub 2} double layer cathodes (DLCs) were prepared with a thin highly active LiCoO{sub 2}-layer by a new double layer tape casting/sintering procedure. The resulting metallic porous precursor plates were mounted into the MCFC and heated up by a special procedure to form LiCoO{sub 2} from air, Co and Li{sub 2}CO{sub 3} in a solid/gas reaction. MCFCs with highly active NiO/LiCoO{sub 2}-DLCs can operate over prolonged periods of time with a Ni-precipitation which is 10% lower than one finds with state of the art NiO cathodes. According to LiCoO{sub 2}-cathodes have theoretical life times of more than 100 000 hours at nonpressurized conditions. MCFCs with new NiO/LiCoO{sub 2} double layer cathodes (DLC) were investigated with regard to variable parameters of their microstructure. From the agglomerate model of the porous MCFC cathode, the dependence of the polarization resistance from the radius of the agglomerates and the inner agglomerate surface area was calculated.

  5. Determination of potential doubling times in human melanoma cell cultures subjected to irradiation and/or hyperthermia by flow cytometry

    SciTech Connect

    Zoelzer, F.; treffer, C.; Devi, P.U.

    1994-06-01

    The proliferation of human melanoma cells in vitro after irradiation and/or hyperthermia was studied by means of two-parameter flow cytometry. Cultures were incubated with BrdU for 30 min and fixed either immediately or after a delay of several hours. Cells having synthesized DNA were identified with the help of an antibody against BrdU. DNA was stained quantitatively with propidium iodide. In this way the distribution of cells in the phases of cell cycle could be determined and the movement of labeled cells through the phases of the cycle could be analyzed. Experiments in which the cell cycle distribution was studied at 4-h intervals after treatment showed the following: (1) Irradiation (4 Gy X rays) causes the expected G{sub 2} block with a maximum after 12-16 h. The proportion of S-phase cells decreases continually during the first 48 h after treatment. (2) Hyperthermia (1 h, 43{degrees}C) alone or in combination with irradiation causes a delay in S phase. The cells begin to move into G{sub 2} phase only after 12-16 h and accumulate there to some extent. From the progression of labeled cells through the cycle, the duration of S phase could be determined. Experiments and calculations of this kind were done 0, 24 and 48 h after treatment. The duration of S phase was increased only moderately (by 4 h) after irradiation, but a delay of about 30 h occurred after hyperthermia (alone or in combination with X rays). Smaller delays (up to 9 h) were observed 24 and 48 h after treatment. Two different methods were used to calculate potential doubling times. Both of them gave similar results, but a comparison with the actual population doubling times (determined by cell counting) showed that reasonable estimates could be achieved only for the untreated controls. With cultures subjected to irradiation and/or hyperthermia serious discrepancies were observed. 14 refs., 5 figs., 4 tabs.

  6. A dual-targeting triplebody mediates preferential redirected lysis of antigen double-positive over single-positive leukemic cells

    PubMed Central

    Schubert, Ingo; Saul, Domenica; Nowecki, Stefanie; Mackensen, Andreas; Fey, Georg H; Oduncu, Fuat S

    2014-01-01

    The single-chain triplebody HLA-ds16-hu19 consists of three single-chain Fv (scFv) antibody fragments connected in a single polypeptide chain. This protein with dual-targeting capacity mediated preferential lysis of antigen double-positive (dp) over single-positive (sp) leukemic cells by recruitment of natural killer (NK) cells as effectors. The two distal scFv modules were specific for the histocompatibility protein HLA-DR and the lymphoid antigen CD19, the central one for the Fc gamma receptor CD16. In antibody-dependent cellular cytotoxicity (ADCC) experiments with a mixture of leukemic target cells comprising both HLA-DR sp HuT-78 or Kasumi-1 cells and (HLA-DR plus CD19) dp SEM cells, the triplebody mediated preferential lysis of the dp cells even when the sp cells were present in ≤20-fold numerical excess. The triplebody promoted equal lysis of SEM cells at 2.5-fold and 19.5-fold lower concentrations than the parental antibodies specific for HLA-DR and CD19, respectively. Finally, the triplebody also eliminated primary leukemic cells at lower concentrations than an equimolar mixture of bispecific single-chain Fv fragments (bsscFvs) separately addressing each target antigen (hu19-ds16 and HLA-ds16). The increased selectivity of targeting and the preferential lysis of dp over sp cells achieved by dual-targeting open attractive new perspectives for the use of dual-targeting agents in cancer therapy. PMID:24135631

  7. p53 modulates homologous recombination at I-SceI-induced double-strand breaks through cell-cycle regulation.

    PubMed

    Rieckmann, T; Kriegs, M; Nitsch, L; Hoffer, K; Rohaly, G; Kocher, S; Petersen, C; Dikomey, E; Dornreiter, I; Dahm-Daphi, J

    2013-02-21

    Inhibition of homologous recombination (HR) is believed to be a transactivation-independent function of p53 that protects from genetic instability. Misrepair by HR can lead to genetic alterations such as translocations, duplications, insertions and loss of heterozygosity, which all bear the risk of driving oncogenic transformation. Regulation of HR by wild-type p53 (wtp53) should prevent these genomic rearrangements. Mutation of p53 is a frequent event during carcinogenesis. In particular, dominant-negative mutants inhibiting wtp53 expressed from the unperturbed allel can drive oncogenic transformation by disrupting the p53-dependent anticancer barrier. Here, we asked whether the hot spot mutants R175H and R273H relax HR control in p53-proficient cells. Utilizing an I-SceI-based reporter assay, we observed a moderate (1.5 × ) stimulation of HR upon expression of the mutant proteins in p53-proficient CV-1, but not in p53-deficient H1299 cells. Importantly, the stimulatory effect was exactly paralleled by an increase in the number of HR competent S- and G2-phase cells, which can well explain the enhanced recombination frequencies. Furthermore, the impact on HR exerted by the transactivation domain double-mutant L22Q/W23S and mutant R273P, both of which were reported to regulate HR independently of G1-arrest execution, is also exactly mirrored by cell-cycle behavior. These results are in contrast to previous concepts stating that the transactivation-independent impact of p53 on HR is a general phenomenon valid for replication-associated and also for directly induced double-strand break. Our data strongly suggest that the latter is largely mediated by cell-cycle regulation, a classical transactivation-dependent function of p53. PMID:22484423

  8. CXCR5 is critically involved in progression of lupus through regulation of B cell and double-negative T cell trafficking.

    PubMed

    Wiener, A; Schippers, A; Wagner, N; Tacke, F; Ostendorf, T; Honke, N; Tenbrock, K; Ohl, K

    2016-07-01

    The recruitment of immune cells to sites of tissue inflammation is orchestrated by chemokine/chemokine receptor networks. Among these, the CXCL13/CXCR5 axis is thought to be involved critically in systemic lupus erythematosus (SLE) and lupus nephritis pathogenesis. Beyond B cell abnormalities, another hallmark of SLE disease is the occurrence of aberrant T cell responses. In particular, double-negative (DN) T cells are expanded in the peripheral blood of patients with SLE and in lupus-prone mice. DN T cells induce immunoglobulin production, secrete proinflammatory cytokines and infiltrate inflamed tissue, including kidneys. We aimed to investigate how CXCR5 deficiency changes immune cell trafficking in murine lupus. We therefore crossed CXCR5(-/-) mice with B6/lpr mice, a well-established murine lupus model. B cell numbers and B cellular immune responses were diminished in CXCR5-deficient B6/lpr mice. In addition, we observed reduced accumulation of DN T cells in spleen and lymph nodes, paralleled by reduced splenomegaly and lymphadenopathy. In-vivo migration assays revealed reduced migration of CXCR5-deficient DN T cells into lymph nodes, and ex-vivo-activated CXCR5-deficient DN T cells failed to infiltrate kidneys of recipients. Moreover, DN T cells and B cells of CXCR5-deficient B6/lpr mice failed to migrate towards CXCL13 in vitro. We propose that CXCR5 is involved critically in B cell trafficking and germinal cell (GC) formation in murine lupus and in guiding pathogenic DN T cells into lymphoid organs and kidneys, and we therefore describe new pathomechanisms for the CXCL13/CXCR5 axis in SLE. PMID:26990531

  9. Infected and apoptotic cells in the IBDV-infected bursa of Fabricius, studied by double-labelling techniques.

    PubMed

    Nieper, H; Teifke, J P; Jungmann, A; Lohr, C V; Muller, H

    1999-06-01

    Infections of young chickens with infectious bursal disease virus (IBDV) result in depletion of lymphoid cells of the bursa of Fabricius (BF) due to necrosis and apoptotic processes. Interactions between IBDV and lymphoid cells were investigated by labelling paraffin-embedded tissue sections of infected BF with combinations of either immunohistochemistry (IHC), in situ hybridization (ISH) or in situ TUNEL reaction (IST). With regard to specificity and sensitivity, results of ISH were comparable to those of IHC. By double-labelling it was shown, for the first time, that viral antigen was present in most of the apoptotic cells. This suggests that IBDV may be directly involved in the induction of the apoptotic process. However, some cells also showed either viral antigen or DNA fragmentation, especially at the early stages of infection. It should be taken into account, therefore, that the apoptotic processes might also be induced by IBDV through indirect interaction between cells. Remarkably, in some of the infected lymphoid cells ISH signals were observed in the nucleolus. PMID:26915384

  10. The double-edge role of B cells in mediating antitumor T-cell immunity: Pharmacological strategies for cancer immunotherapy.

    PubMed

    Wang, Jing-Zhang; Zhang, Yu-Hua; Guo, Xin-Hua; Zhang, Hong-Yan; Zhang, Yuan

    2016-07-01

    Emerging evidence reveals the controversial role of B cells in antitumor immunity, but the underlying mechanisms have to be explored. Three latest articles published in the issue 521 of Nature in 2015 reconfirmed the puzzling topic and put forward some explanations of how B cells regulate antitumor T-cell responses both positively and negatively. This paper attempts to demonstrate that different B-cell subpopulations have distinct immunological properties and that they are involved in either antitumor responses or immunosuppression. Recent studies supporting the positive and negative roles of B cells in tumor development were summarized comprehensively. Several specific B-cell subpopulations, such as IgG(+), IgA(+), IL-10(+), and regulatory B cells, were described in detail. The mechanisms underlying the controversial B-cell effects were mainly attributed to different B-cell subpopulations, different B-cell-derived cytokines, direct B cell-T cell interaction, different cancer categories, and different malignant stages, and the immunological interaction between B cells and T cells is mediated by dendritic cells. Promising B-cell-based antitumor strategies were proposed and novel B-cell regulators were summarized to present interesting therapeutic targets. Future investigations are needed to make sure that B-cell-based pharmacological strategies benefit cancer immunotherapy substantially. PMID:27111515

  11. Mushroom acidic glycosphingolipid induction of cytokine secretion from murine T cells and proliferation of NK1.1 {alpha}/{beta} TCR-double positive cells in vitro

    SciTech Connect

    Nozaki, Hirofumi; Itonori, Saki; Sugita, Mutsumi; Nakamura, Kimihide; Ohba, Kiyoshi; Suzuki, Akemi; Kushi, Yasunori

    2008-08-29

    Interferon (IFN)-{gamma} and interleukin (IL)-4 regulate many types of immune responses. Here we report that acidic glycosphingolipids (AGLs) of Hypsizigus marmoreus and Pleurotus eryngii induced secretion of IFN- {gamma} and IL-4 from T cells in a CD11c-positive cell-dependent manner similar to that of {alpha}-galactosylceramide ({alpha}-GalCer) and isoglobotriaosylceramide (iGb3), although activated T cells by AGLs showed less secretion of cytokine than those activated by {alpha}-GalCer. In addition, stimulation of these mushroom AGLs induced proliferation of NK1.1 {alpha}/{beta} TCR-double positive cells in splenocytes. Administration of a mixture of {alpha}-GalCer and AGLs affected the stimulation of {alpha}-GalCer and generally induced a subtle Th1 bias for splenocytes but induced an extreme Th2 bias for thymocytes. These results suggested that edible mushroom AGLs contribute to immunomodulation.

  12. Investigation of the double exponential in the current-voltage characteristics of silicon solar cells. [proton irradiation effects on ATS 1 cells

    NASA Technical Reports Server (NTRS)

    Wolf, M.; Noel, G. T.; Stirn, R. J.

    1977-01-01

    Difficulties in relating observed current-voltage characteristics of individual silicon solar cells to their physical and material parameters were underscored by the unexpected large changes in the current-voltage characteristics telemetered back from solar cells on the ATS-1 spacecraft during their first year in synchronous orbit. Depletion region recombination was studied in cells exhibiting a clear double-exponential dark characteristic by subjecting the cells to proton irradiation. A significant change in the saturation current, an effect included in the Sah, Noyce, Shockley formulation of diode current resulting from recombination in the depletion region, was caused by the introduction of shallow levels in the depletion region by the proton irradiation. This saturation current is not attributable only to diffusion current from outside the depletion region and only its temperature dependence can clarify its origin. The current associated with the introduction of deep-lying levels did not change significantly in these experiments.

  13. The fabrication of a double-layer atom chip with through silicon vias for an ultra-high-vacuum cell

    NASA Astrophysics Data System (ADS)

    Chuang, Ho-Chiao; Lin, Yun-Siang; Lin, Yu-Hsin; Huang, Chi-Sheng

    2014-04-01

    This study presents a double-layer atom chip that provides users with increased diversity in the design of the wire patterns and flexibility in the design of the magnetic field. It is more convenient for use in atomic physics experiments. A negative photoresist, SU-8, was used as the insulating layer between the upper and bottom copper wires. The electrical measurement results show that the upper and bottom wires with a width of 100 µm can sustain a 6 A current without burnout. Another focus of this study is the double-layer atom chips integrated with the through silicon via (TSV) technique, and anodically bonded to a Pyrex glass cell, which makes it a desired vacuum chamber for atomic physics experiments. Thus, the bonded glass cell not only significantly reduces the overall size of the ultra-high-vacuum (UHV) chamber but also conducts the high current from the backside to the front side of the atom chip via the TSV under UHV (9.5 × 10-10 Torr). The TSVs with a diameter of 70 µm were etched through by the inductively coupled plasma ion etching and filled by the bottom-up copper electroplating method. During the anodic bonding process, the electroplated copper wires and TSVs on atom chips also need to pass the examination of the required bonding temperature of 250 °C, under an applied voltage of 1000 V. Finally, the UHV test of the double-layer atom chips with TSVs at room temperature can be reached at 9.5 × 10-10 Torr, thus satisfying the requirements of atomic physics experiments under an UHV environment.

  14. Graft derived cells with double nuclei in the penumbral region of experimental brain trauma.

    PubMed

    Horváth, Eszter M; Lacza, Zsombor; Csordás, Attila; Szabó, Csaba; Kollai, Márk; Busija, David W

    2006-04-01

    Recent in vitro studies showed that stem cells might fuse with mature cells or each other; however, there is no in vivo evidence for this phenomenon in the cerebral cortex. Our goal was to find evidence for cell fusion in a model of traumatic brain injury followed by grafting of embryonic cortical cells. Cold lesion protocol was applied to induce lesion of the motor cortex in adult male rats. Six days later we grafted a suspension of freshly isolated rat brain cortical cells of early embryonic stage (E14) into the penumbra area of the lesion. The grafted cell nuclei were labelled with bromodeoxyuridine (BrDU). Six days after transplantation 4,328 BrDU positive cells were observed in nine animals. 89.5% of these cells had cytoplasmic staining probably representing dead or phagocyted grafted cells. Ten percent of surviving BrDU positive cells had only one BrDU positive nucleus and negative cytoplasm, while 0.5% had two distinct nuclei, one was unlabelled and one was BrDU positive. These cells were similar in appearance and size to the astrocytes in the vicinity and expressed the astocyte specific glial fibrillaly acidic protein. Thus, these cells showed a possible sign of cell fusion in the penumbral region of the injured brain. PMID:16377084

  15. RAG-mediated DNA double-strand breaks activate a cell type-specific checkpoint to inhibit pre-B cell receptor signals.

    PubMed

    Bednarski, Jeffrey J; Pandey, Ruchi; Schulte, Emily; White, Lynn S; Chen, Bo-Ruei; Sandoval, Gabriel J; Kohyama, Masako; Haldar, Malay; Nickless, Andrew; Trott, Amanda; Cheng, Genhong; Murphy, Kenneth M; Bassing, Craig H; Payton, Jacqueline E; Sleckman, Barry P

    2016-02-01

    DNA double-strand breaks (DSBs) activate a canonical DNA damage response, including highly conserved cell cycle checkpoint pathways that prevent cells with DSBs from progressing through the cell cycle. In developing B cells, pre-B cell receptor (pre-BCR) signals initiate immunoglobulin light (Igl) chain gene assembly, leading to RAG-mediated DNA DSBs. The pre-BCR also promotes cell cycle entry, which could cause aberrant DSB repair and genome instability in pre-B cells. Here, we show that RAG DSBs inhibit pre-BCR signals through the ATM- and NF-κB2-dependent induction of SPIC, a hematopoietic-specific transcriptional repressor. SPIC inhibits expression of the SYK tyrosine kinase and BLNK adaptor, resulting in suppression of pre-BCR signaling. This regulatory circuit prevents the pre-BCR from inducing additional Igl chain gene rearrangements and driving pre-B cells with RAG DSBs into cycle. We propose that pre-B cells toggle between pre-BCR signals and a RAG DSB-dependent checkpoint to maintain genome stability while iteratively assembling Igl chain genes. PMID:26834154

  16. Co-culturing with High-Charge and Energy Particle Irradiated Cells Increases Mutagenic Joining of Enzymatically Induced DNA Double-Strand Breaks in Nonirradiated Cells.

    PubMed

    Li, Zhentian; Doho, Gregory; Zheng, Xuan; Jella, Kishore Kumar; Li, Shuyi; Wang, Ya; Dynan, William S

    2015-09-01

    Cell populations that have been exposed to high-charge and energy (HZE) particle radiation, and then challenged by expression of a rare-cutting nuclease, show an increased frequency of deletions and translocations originating at the enzyme cut sites. Here, we examine whether this effect also occurs in nonirradiated cells that have been co-cultured with irradiated cells. Human cells were irradiated with 0.3-1.0 Gy of either 600 MeV/u (56)Fe or 1,000 MeV/u (48)Ti ions or with 0.3-3.0 Gy of 320 kV X rays. These were co-cultured with I-SceI-expressing reporter cells at intervals up to 21 days postirradiation. Co-culture with HZE-irradiated cells led to an increase in the frequency of I-SceI-stimulated translocations and deletions in the nonirradiated cells. The effect size was similar to that seen previously in directly irradiated populations (maximum effect in bystander cells of 1.7- to 4-fold depending on ion and end point). The effect was not observed when X-ray-irradiated cells were co-cultured with nonirradiated cells, but was correlated with an increase in γ-H2AX foci-positive cells in the nonirradiated population, suggesting the presence of genomic stress. Transcriptional profiling of a directly irradiated cell population showed that many genes for cytokines and other secretory proteins were persistently upregulated, but their induction was not well correlated with functional effects on repair in co-cultured cells, suggesting that this transcriptional response alone is not sufficient to evoke the effect. The finding that HZE-irradiated cells influence the DNA double-strand break repair fidelity in their nonirradiated neighbors has implications for risk in the space radiation environment. PMID:26284422

  17. IKKβ regulates the repair of DNA double-strand breaks induced by ionizing radiation in MCF-7 breast cancer cells.

    PubMed

    Wu, Lixian; Shao, Lijian; An, Ningfei; Wang, Junru; Pazhanisamy, Senthil; Feng, Wei; Hauer-Jensen, Martin; Miyamoto, Shigeki; Zhou, Daohong

    2011-01-01

    Activation of the IKK-NFκB pathway increases the resistance of cancer cells to ionizing radiation (IR). This effect has been largely attributed to the induction of anti-apoptotic proteins by NFκB. Since efficient repair of DNA double strand breaks (DSBs) is required for the clonogenic survival of irradiated cells, we investigated if activation of the IKK-NFκB pathway also regulates DSB repair to promote cell survival after IR. We found that inhibition of the IKK-NFκB pathway with a specific IKKβ inhibitor significantly reduced the repair of IR-induced DSBs in MCF-7 cells. The repair of DSBs was also significantly inhibited by silencing IKKβ expression with IKKβ shRNA. However, down-regulation of IKKα expression with IKKα shRNA had no significant effect on the repair of IR-induced DSBs. Similar findings were also observed in IKKα and/or IKKβ knockout mouse embryonic fibroblasts (MEFs). More importantly, inhibition of IKKβ with an inhibitor or down-regulation of IKKβ with IKKβ shRNA sensitized MCF-7 cells to IR-induced clonogenic cell death. DSB repair function and resistance to IR were completely restored by IKKβ reconstitution in IKKβ-knockdown MCF-7 cells. These findings demonstrate that IKKβ can regulate the repair of DSBs, a previously undescribed and important IKKβ kinase function; and inhibition of DSB repair may contribute to cance cell radiosensitization induced by IKKβ inhibition. As such, specific inhibition of IKKβ may represents a more effective approach to sensitize cancer cells to radiotherapy. PMID:21490922

  18. Concomitant charge-density-wave and unit-cell-doubling structural transitions in Dy5Ir4Si10

    NASA Astrophysics Data System (ADS)

    Lee, M. H.; Chen, C. H.; Tseng, C. M.; Lue, C. S.; Kuo, Y. K.; Yang, H. D.; Chu, M.-W.

    2014-05-01

    The tetragonal rare-earth transition-metal silicide system with three-dimensional crystallographic structure R5T4Si10, where R is Dy, Ho, Er, Tm, and Lu, and T=Ir and Rh, has been shown to exhibit fascinating charge-density-wave (CDW) phase transitions, a phenomenon largely found in otherwise low-dimensional systems. In this study, we report the investigations of CDW in Dy5Ir4Si10 at different temperatures using transmission electron microscopy techniques including electron diffraction and dark-field imaging. Incommensurate superlattice spots along the c axis were observed in the electron-diffraction patterns when the sample was cooled below the CDW transition temperature at ˜208 K. CDW becomes commensurate with further cooling and configurations of CDW dislocations convincingly show that the CDW phase transition is accompanied by a concomitant cell-doubling crystallographic structural phase transition. Intriguingly, the cell-doubling transition is featured by a broken inversion symmetry along the c axis and a disparity in the CDW-modulation vectors with opposite signs, which gives rise to two sets of CDW domains with reversed contrasts. The profound physics underlining this notable domain-contrast behavior is discussed.

  19. Detailed flowfield and surface properties for high Knudsen number planar jet impingement at an inclined flat plate

    NASA Astrophysics Data System (ADS)

    Cai, Chunpei; He, Xin

    2016-05-01

    This paper presents two sets of analytical exact solutions for collisionless gas flows from a planar exit, impinging at an inclined flat plate. These analytical results are obtained by using gaskinetic theories. The first set of solutions are for a diffuse reflective plate surface, and the other set of solutions are for a specular reflective plate surface. A virtual nozzle exit is adopted to aid analyzing the specular reflective plate scenario. New formulas for plate surface properties, including velocity slips, pressure, shear stress, and heat flux distributions, are provided. For both problems, the flowfield exact solutions are investigated as well. Numerical simulations with the direct simulation Monte Carlo method are performed to validate these new analytical results, and good agreement is obtained for flows with high Knudsen numbers. The results consider effects from many factors, such as the plate inclination angle, geometry ratios, and exit gas and plate properties (such as exit gas bulk density, gas speed ratio, and exit gas and plate temperatures). Compared with past work, these new solutions are more comprehensive and practical. The results also illustrate that if the plate is quite close to the nozzle exit, it is improper to adopt the traditional treatments of a point source and a simple cosine function.

  20. A dynamic response model for pressure sensors in continuum and high Knudsen number flows with large temperature gradients

    NASA Technical Reports Server (NTRS)

    Whitmore, Stephen A.; Petersen, Brian J.; Scott, David D.

    1996-01-01

    This paper develops a dynamic model for pressure sensors in continuum and rarefied flows with longitudinal temperature gradients. The model was developed from the unsteady Navier-Stokes momentum, energy, and continuity equations and was linearized using small perturbations. The energy equation was decoupled from momentum and continuity assuming a polytropic flow process. Rarefied flow conditions were accounted for using a slip flow boundary condition at the tubing wall. The equations were radially averaged and solved assuming gas properties remain constant along a small tubing element. This fundamental solution was used as a building block for arbitrary geometries where fluid properties may also vary longitudinally in the tube. The problem was solved recursively starting at the transducer and working upstream in the tube. Dynamic frequency response tests were performed for continuum flow conditions in the presence of temperature gradients. These tests validated the recursive formulation of the model. Model steady-state behavior was analyzed using the final value theorem. Tests were performed for rarefied flow conditions and compared to the model steady-state response to evaluate the regime of applicability. Model comparisons were excellent for Knudsen numbers up to 0.6. Beyond this point, molecular affects caused model analyses to become inaccurate.

  1. EXCISION OF GIANT CELL TUMOR OF TENDON SHEATH WITH BONE INVOLVEMENT BY MEANS OF DOUBLE ACCESS APPROACH: CASE REPORT

    PubMed Central

    Alves, Marcelo de Pinho Teixeira

    2015-01-01

    Giant cell tumors of the tendon sheath are common lesions and are the second most frequent tumors in the hand, after synovial cysts. They are diagnosed by means of clinical examination and complementary examinations (simple radiography and magnetic resonance). Erosion and invasion of the phalangeal bone affected may be seen on radiological examination. Magnetic resonance may show a “fluorescent or radiant effect” may be observed, caused by the high quantity of hemosiderin inside the tumor. Surgical treatment is the commonest practice, and complete excision is important for avoiding recurrence of the tumor, especially when bone invasion is observed on imaging examinations, which is generally related to greater tumor recurrence. In this paper, a case of a giant cell tumor of the tendon sheath in the middle phalanx of the third finger of a 45-year-old female patient is presented. This was successfully treated by means of surgery using a double access approach (dorsal and volar). PMID:27026996

  2. T-Regulatory Cell and CD3 Depleted Double Umbilical Cord Blood Transplantation in Hematologic Malignancies

    ClinicalTrials.gov

    2014-03-04

    Hematologic Malignancy; Acute Myeloid Leukemia; Acute Lymphocytic Leukemia; Chronic Myelogenous Leukemia in Blast Crisis; Anemia, Refractory, With Excess of Blasts; Chronic Myeloproliferative Disease; Chronic Lymphocytic Leukemia; Small Lymphocytic Lymphoma; Marginal Zone B-cell Lymphoma; Follicular Lymphoma; Lymphoplasmacytic Lymphoma; Mantle-Cell Lymphoma; Prolymphocytic Lymphoma; Large Cell Non-Hodgkin's Lymphoma; Lymphoblastic Lymphoma; Burkitt's Lymphoma; High Grade Non-Hodgkin's Lymphoma

  3. Atrazine Triggers DNA Damage Response and Induces DNA Double-Strand Breaks in MCF-10A Cells

    PubMed Central

    Huang, Peixin; Yang, John; Ning, Jie; Wang, Michael; Song, Qisheng

    2015-01-01

    Atrazine, a pre-emergent herbicide in the chloro-s-triazine family, has been widely used in crop lands and often detected in agriculture watersheds, which is considered as a potential threat to human health. Although atrazine and its metabolites showed an elevated incidence of mammary tumors in female Sprague–Dawley (SD) rats, no molecular evidence was found relevant to its carcinogenesis in humans. This study aims to determine whether atrazine could induce the expression of DNA damage response-related proteins in normal human breast epithelial cells (MCF-10A) and to examine the cytotoxicity of atrazine at a molecular level. Our results indicate that a short-term exposure of MCF-10A to an environmentally-detectable concentration of atrazine (0.1 µg/mL) significantly increased the expression of tumor necrosis factor receptor-1 (TNFR1) and phosphorylated Rad17 in the cells. Atrazine treatment increased H2AX phosphorylation (γH2AX) and the formation of γH2AX foci in the nuclei of MCF-10A cells. Atrazine also sequentially elevated DNA damage checkpoint proteins of ATM- and RAD3-related (ATR), ATRIP and phospho-Chk1, suggesting that atrazine could induce DNA double-strand breaks and trigger the DNA damage response ATR-Chk1 pathway in MCF-10A cells. Further investigations are needed to determine whether atrazine-triggered DNA double-strand breaks and DNA damage response ATR-Chk1 pathway occur in vivo. PMID:26114388

  4. Atrazine Triggers DNA Damage Response and Induces DNA Double-Strand Breaks in MCF-10A Cells.

    PubMed

    Huang, Peixin; Yang, John; Ning, Jie; Wang, Michael; Song, Qisheng

    2015-01-01

    Atrazine, a pre-emergent herbicide in the chloro-s-triazine family, has been widely used in crop lands and often detected in agriculture watersheds, which is considered as a potential threat to human health. Although atrazine and its metabolites showed an elevated incidence of mammary tumors in female Sprague-Dawley (SD) rats, no molecular evidence was found relevant to its carcinogenesis in humans. This study aims to determine whether atrazine could induce the expression of DNA damage response-related proteins in normal human breast epithelial cells (MCF-10A) and to examine the cytotoxicity of atrazine at a molecular level. Our results indicate that a short-term exposure of MCF-10A to an environmentally-detectable concentration of atrazine (0.1 µg/mL) significantly increased the expression of tumor necrosis factor receptor-1 (TNFR1) and phosphorylated Rad17 in the cells. Atrazine treatment increased H2AX phosphorylation (γH2AX) and the formation of γH2AX foci in the nuclei of MCF-10A cells. Atrazine also sequentially elevated DNA damage checkpoint proteins of ATM- and RAD3-related (ATR), ATRIP and phospho-Chk1, suggesting that atrazine could induce DNA double-strand breaks and trigger the DNA damage response ATR-Chk1 pathway in MCF-10A cells. Further investigations are needed to determine whether atrazine-triggered DNA double-strand breaks and DNA damage response ATR-Chk1 pathway occur in vivo. PMID:26114388

  5. Spindle cell carcinoma of the tongue combined with double primary cancer of the thyroid gland: a case report.

    PubMed

    Kim, Tae-Hwan; Kim, Chul-Hwan

    2016-08-01

    Spindle cell carcinoma (SpCC) is referred to as a variant of oral squamous cell carcinoma. It is also known as "sarcomatoid squamous cell carcinoma" because it consists of normal squamous carcinoma cells with spindle-shaped cells that appear similar to a sarcoma. The term, "second primary tumor" (SPT) or "double primary tumor", is proposed for a second tumor that develops independently from the first. SPTs can present as either synchronous or metachronous lesions. Synchronous SPTs are defined as tumors occurring simultaneously or within 6 months after the first tumor. The patient in this case, whose primary tumor was in the tongue, was diagnosed with SpCC with metastases to both neck lymph nodes. This case also exhibited a second primary cancer as a synchronous lesion in the thyroid gland, which is uncommon. All carcinomas, both in the tongue and thyroid gland, were removed surgically, and especially in the tongue, an anterolateral thigh free flap was performed successfully to replace the defect. PMID:27595090

  6. Spindle cell carcinoma of the tongue combined with double primary cancer of the thyroid gland: a case report

    PubMed Central

    2016-01-01

    Spindle cell carcinoma (SpCC) is referred to as a variant of oral squamous cell carcinoma. It is also known as "sarcomatoid squamous cell carcinoma" because it consists of normal squamous carcinoma cells with spindle-shaped cells that appear similar to a sarcoma. The term, "second primary tumor" (SPT) or "double primary tumor", is proposed for a second tumor that develops independently from the first. SPTs can present as either synchronous or metachronous lesions. Synchronous SPTs are defined as tumors occurring simultaneously or within 6 months after the first tumor. The patient in this case, whose primary tumor was in the tongue, was diagnosed with SpCC with metastases to both neck lymph nodes. This case also exhibited a second primary cancer as a synchronous lesion in the thyroid gland, which is uncommon. All carcinomas, both in the tongue and thyroid gland, were removed surgically, and especially in the tongue, an anterolateral thigh free flap was performed successfully to replace the defect. PMID:27595090

  7. Selective Tumor Cell Inhibition Effect of Ni-Ti Layered Double Hydroxides Thin Films Driven by the Reversed pH Gradients of Tumor Cells.

    PubMed

    Wang, Donghui; Ge, Naijian; Li, Jinhua; Qiao, Yuqin; Zhu, Hongqin; Liu, Xuanyong

    2015-04-22

    Nitinol is widely fabricated as stents for the palliation treatment of many kinds of cancers. It is of great importance to develop nitinol stents with selective tumor cell inhibition effects. In this work, a series of pH sensitive films composed of Ni(OH)2 and Ni-Ti layered double hydroxide (Ni-Ti LDH) with different Ni/Ti ratios were prepared on the surface of nitinol via hydrothermal treatment. The films with specific Ni/Ti ratios would release a large amount of nickel ions under acidic environments but were relatively stable in neutral or weak alkaline medium. Cell viability tests showed that the films can effectively inhibit the growth of cancer cells but have little adverse effects to normal cells. Besides, extraordinarily high intracellular nickel content and reactive oxygen species (ROS) level were found in cancer cells, indicating the death of cancer cells may be induced by the excessive intake of nickel ions. Such selective cancer cell inhibition effect of the films is supposed to relate with the reversed pH gradients of tumor cells. PMID:25825800

  8. Prevention of DNA Double-Strand Breaks Induced by Radioiodide-131I in FRTL-5 Thyroid Cells

    PubMed Central

    Okunyan, Armen; Rivina, Yelena; Cannon, Sophie; Hogen, Victor

    2011-01-01

    Radioiodine-131 released from nuclear reactor accidents has dramatically increased the incidence of papillary thyroid cancer in exposed individuals. The deposition of ionizing radiation in cells results in double-strand DNA breaks (DSB) at fragile sites, and this early event can generate oncogenic rearrangements that eventually cause cancer. The aims of this study were to develop a method to show DNA DSBs induced by 131I in thyroid cells; to test monovalent anions that are transported by the sodium/iodide symporter to determine whether they prevent 131I-induced DSB; and to test other radioprotective agents for their effect on irradiated thyroid cells. Rat FRTL-5 thyroid cells were incubated with 131I. DSBs were measured by nuclear immunofluorescence using antibodies to p53-binding protein 1 or γH2AX. Incubation with 1–10 μCi 131I per milliliter for 90 min resulted in a dose-related increase of DSBs; the number of DSBs increased from a baseline of 4–15% before radiation to 65–90% after radiation. GH3 or CHO cells that do not transport iodide did not develop DSBs when incubated with 131I. Incubation with 20–100 μm iodide or thiocyanate markedly attenuated DSBs. Perchlorate was about 6 times more potent than iodide or thiocyanate. The effects of the anions were much greater when each was added 30–120 min before the 131I. Two natural organic compounds recently shown to provide radiation protection partially prevented DSBs caused by 131I and had an additive effect with perchlorate. In conclusion, we developed a thyroid cell model to quantify the mitogenic effect of 131I. 131I causes DNA DSBs in FRTL-5 cells and had no effect on cells that do not transport iodide. Perchlorate, iodide, and thiocyanate protect against DSBs induced by 131I. PMID:21190956

  9. Isolation of Double Negative αβ T Cells from the Kidney

    PubMed Central

    Martina, Maria N.; Bandapalle, Samantha; Rabb, Hamid; Hamad, Abdel R.

    2014-01-01

    There is currently no standard protocol for the isolation of DN T cells from the non-lymphoid tissues despite their increasingly reported involvement in various immune responses. DN T cells are a unique immune cell type that has been implicated in regulating immune and autoimmune responses and tolerance to allotransplants1-6. DN T cells are, however, rare in peripheral blood and secondary lymphoid organs (spleen and lymph nodes), but are major residents of the normal kidney. Very little is known about their pathophysiologic function7 due to their paucity in the periphery. We recently described a comprehensive phenotypic and functional analysis of this population in the kidney8 in steady state and during ischemia reperfusion injury. Analysis of DN T cell function will be greatly enhanced by developing a protocol for their isolation from the kidney. Here, we describe a novel protocol that allows isolation of highly pure ab CD4+ CD8+ T cells and DN T cells from the murine kidney. Briefly, we digest kidney tissue using collagenase and isolate kidney mononuclear cells (KMNC) by density gradient. This is followed by two steps to enrich hematopoietic T cells from 3% to 70% from KMNC. The first step consists of a positive selection of hematopoietic cells using a CD45+ isolation kit. In the second step, DN T cells are negatively isolated by removal of non-desired cells using CD4, CD8, and MHC class II monoclonal antibodies and CD1d α-galcer tetramer. This strategy leads to a population of more than 90% pure DN T cells. Surface staining with the above mentioned antibodies followed by FACs analysis is used to confirm purity. PMID:24893925

  10. A double suicide gene system driven by vascular endothelial growth factor promoter selectively kills human hepatocellular carcinoma cells

    PubMed Central

    WU, KAI; YANG, LIUCHENG; HUANG, ZONGHAI; ZHAO, HAIJUN; WANG, JIANJUN; XU, SHUAI

    2016-01-01

    The aim of the present study was to investigate the selective killing effect on hepatocellular carcinoma (HCC) cells of an adenovirus (Ad)-mediated cytosine deaminase (CD) in combination with thymidine kinase (TK) suicide gene system, driven by the vascular endothelial growth factor promoter (VEGFp), in vitro and in vivo. A double suicide gene system with VEGFp, named Ad-VEGFp-CDglyTK, was constructed and transfected into human HCC cells (BEL-7402 or HepG2; the latter cell type is deficient in VEGF) and human umbilical vein vascular endothelial cells (HUVEC). Green fluorescent protein expression was detected by fluoroscopy to verify transfection efficiency, and CDglyTK gene expression was detected by reverse transcription-polymerase chain reaction (PCR). The selective killing effect of Ad-VEGFp-CDglyTK was measured by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay and flow cytometry (FCM) in vitro and by xenograft studies in vivo. PCR revealed that the transgenic CDglyTK gene was expressed in BEL-7402 cells and HUVEC, but not in HepG2 cells. The cell survival rate significantly decreased in line with increasing concentrations of the prodrugs, ganciclovir (GCV) alone, 5-fluorocytosine (5-FC) alone or a combination of the two, in HUVEC and BEL-7402 cells with the transfected CDglyTK gene, but not in untransfected HUVEC or BEL-7402 cells, or in transfected or untransfected HepG2 cells. This result was additionally confirmed by FCM. GCV and 5-FC inhibited the HUVEC and BEL-7402 cells containing the transfected CDglyTK gene and also inhibited adjacent unmodified cells via the ‘bystander effect’. No similar results were observed in HepG2 cells. Compared with the control group, tumors with the transfected CDglyTK gene were smaller and the microvessel density of the tumor tissue was significantly decreased. It was concluded that a combination TK/GCV and CD/5-FC suicide gene system driven by VEGFp may provide a promising treatment strategy for HCC

  11. GaInNAs/Ge (1.10/0.67 eV) double-junction solar cell grown by metalorganic chemical vapor deposition for high efficiency four-junction solar cell application

    NASA Astrophysics Data System (ADS)

    Zhang, Xiaobin; Chen, Bingzhen; Pan, Xu; Wang, Lei; Ma, Difei; Zhang, Yang; Yang, Cuibai; Wang, Zhiyong

    2015-12-01

    GaInNAs materials with narrow bandgaps of 1.10 eV have been grown on a Ge substrate by metalorganic chemical vapor deposition to fabricate GaInNAs/Ge (1.10/0.67 eV) double-junction solar cells. We have studied the photovoltaic characteristics and the external quantum efficiencies of the double-junction cells with various annealing conditions and different GaInNAs base layer thicknesses. The best external quantum efficiency is obtained from the double-junction cell with a 1170 nm thick GaInNAs base layer annealed at 675 °C for 30 min. Under AM1.5G illumination, the best double-junction cell has a short circuit current density (J SC) as 23.63 mA cm-2, which is dominated by the J SC of the GaInNAs subcell.

  12. Oncogenic ras-driven cancer cell vesiculation leads to emission of double-stranded DNA capable of interacting with target cells.

    PubMed

    Lee, Tae Hoon; Chennakrishnaiah, Shilpa; Audemard, Eric; Montermini, Laura; Meehan, Brian; Rak, Janusz

    2014-08-22

    Cell free DNA is often regarded as a source of genetic cancer biomarkers, but the related mechanisms of DNA release, composition and biological activity remain unclear. Here we show that rat epithelial cell transformation by the human H-ras oncogene leads to an increase in production of small, exosomal-like extracellular vesicles by viable cancer cells. These EVs contain chromatin-associated double-stranded DNA fragments covering the entire host genome, including full-length H-ras. Oncogenic N-ras and SV40LT sequences were also found in EVs emitted from spontaneous mouse brain tumor cells. Disruption of acidic sphingomyelinase and the p53/Rb pathway did not block emission of EV-related oncogenic DNA. Exposure of non-transformed RAT-1 cells to EVs containing mutant H-ras DNA led to the uptake and retention of this material for an extended (30days) but transient period of time, and stimulated cell proliferation. Thus, our study suggests that H-ras-mediated transformation stimulates vesicular emission of this histone-bound oncogene, which may interact with non-transformed cells. PMID:25086355

  13. A77 1726, the active metabolite of leflunomide, attenuates lupus nephritis by promoting the development of regulatory T cells and inhibiting IL-17-producing double negative T cells.

    PubMed

    Qiao, Guilin; Yang, Lifen; Li, Zhenping; Williams, James W; Zhang, Jian

    2015-04-01

    Lupus nephritis (LN) is a challenging problem that affects 50% of patients with systemic lupus erythematosus (SLE) without effective therapy. Here, we report that A77 1726, the active metabolite of leflunomide, effectively inhibits development of LN and attenuates the generalized autoimmune features. A77 1726 suppresses the expansion of double negative (DN) T cells, and inhibits T and B cell activation. Intriguingly, A77 1726 treatment significantly increases CD4(+)Foxp3(+) regulatory T cells but suppresses potential "pathogenic" IL-17-producing DN T cells in lymph nodes. In vitro experiment shows that A77 1726 potentiates the conversion of naive CD4(+)CD25(-) T cells into CD4(+)CD25(+)Foxp3(+) inducible regulatory T cells (iTregs) by inhibiting Akt. Taken together, our data indicate that the therapeutic effects of A77 1726 in murine LN are mediated, at least in part, by augmenting iTregs which suppress pathogenic IL-17-producing DN T cells through an Akt-dependent mechanism. PMID:25638413

  14. Depletion of tyrosyl DNA phosphodiesterase 2 activity enhances etoposide-mediated double-strand break formation and cell killing.

    PubMed

    Kont, Yasemin Saygideger; Dutta, Arijit; Mallisetty, Apurva; Mathew, Jeena; Minas, Tsion; Kraus, Christina; Dhopeshwarkar, Priyanka; Kallakury, Bhaskar; Mitra, Sankar; Üren, Aykut; Adhikari, Sanjay

    2016-07-01

    DNA topoisomerase 2 (Top2) poisons, including common anticancer drugs etoposide and doxorubicin kill cancer cells by stabilizing covalent Top2-tyrosyl-DNA 5'-phosphodiester adducts and DNA double-strand breaks (DSBs). Proteolytic degradation of the covalently attached Top2 leaves a 5'-tyrosylated blocked termini which is removed by tyrosyl DNA phosphodiesterase 2 (TDP2), prior to DSB repair through non-homologous end joining (NHEJ). Thus, TDP2 confers resistance of tumor cells to Top2-poisons by repairing such covalent DNA-protein adducts, and its pharmacological inhibition could enhance the efficacy of Top2-poisons. We discovered NSC111041, a selective inhibitor of TDP2, by optimizing a high throughput screening (HTS) assay for TDP2's 5'-tyrosyl phosphodiesterase activity and subsequent validation studies. We found that NSC111041 inhibits TDP2's binding to DNA without getting intercalated into DNA and enhanced etoposide's cytotoxicity synergistically in TDP2-expressing cells but not in TDP2 depleted cells. Furthermore, NSC111041 enhanced formation of etoposide-induced γ-H2AX foci presumably by affecting DSB repair. Immuno-histochemical analysis showed higher TDP2 expression in a sub-set of different type of tumor tissues. These findings underscore the feasibility of clinical use of suitable TDP2 inhibitors in adjuvant therapy with Top2-poisons for a sub-set of cancer patients with high TDP2 expression. PMID:27235629

  15. Post-irradiation chemical processing of DNA damage generates double-strand breaks in cells already engaged in repair

    PubMed Central

    Singh, Satyendra K.; Wang, Minli; Staudt, Christian; Iliakis, George

    2011-01-01

    In cells exposed to ionizing radiation (IR), double-strand breaks (DSBs) form within clustered-damage sites from lesions disrupting the DNA sugar–phosphate backbone. It is commonly assumed that these DSBs form promptly and are immediately detected and processed by the cellular DNA damage response (DDR) apparatus. This assumption is questioned by the observation that after irradiation of naked DNA, a fraction of DSBs forms minutes to hours after exposure as a result of temperature dependent, chemical processing of labile sugar lesions. Excess DSBs also form when IR-exposed cells are processed at 50°C, but have been hitherto considered method-related artifact. Thus, it remains unknown whether DSBs actually develop in cells after IR exposure from chemically labile damage. Here, we show that irradiation of ‘naked’ or chromatin-organized mammalian DNA produces lesions, which evolve to DSBs and add to those promptly induced, after 8–24 h in vitro incubation at 37°C or 50°C. The conversion is more efficient in chromatin-associated DNA, completed within 1 h in cells and delayed in a reducing environment. We conclude that IR generates sugar lesions within clustered-damage sites contributing to DSB formation only after chemical processing, which occurs efficiently at 37°C. This subset of delayed DSBs may challenge DDR, may affect the perceived repair kinetics and requires further characterization. PMID:21745815

  16. BRCA2 and RAD51 promote double-strand break formation and cell death in response to gemcitabine.

    PubMed

    Jones, Rebecca M; Kotsantis, Panagiotis; Stewart, Grant S; Groth, Petra; Petermann, Eva

    2014-10-01

    Replication inhibitors cause replication fork stalling and double-strand breaks (DSB) that result from processing of stalled forks. During recovery from replication blocks, the homologous recombination (HR) factor RAD51 mediates fork restart and DSB repair. HR defects therefore sensitize cells to replication inhibitors, with clear implications for cancer therapy. Gemcitabine is a potent replication inhibitor used to treat cancers with mutations in HR genes such as BRCA2. Here, we investigate why, paradoxically, mutations in HR genes protect cells from killing by gemcitabine. Using DNA replication and DNA damage assays in mammalian cells, we show that even short gemcitabine treatments cause persistent replication inhibition. BRCA2 and RAD51 are recruited to chromatin early after removal of the drug, actively inhibit replication fork progression, and promote the formation of MUS81- and XPF-dependent DSBs that remain unrepaired. Our data suggest that HR intermediates formed at gemcitabine-stalled forks are converted into DSBs and thus contribute to gemcitabine-induced cell death, which could have implications for the treatment response of HR-deficient tumors. PMID:25053826

  17. Inflammation and Inflammatory Cells in Myocardial Infarction and Reperfusion Injury: A Double-Edged Sword

    PubMed Central

    Liu, Jiaqi; Wang, Haijuan; Li, Jun

    2016-01-01

    Myocardial infarction (MI) is the most common cause of cardiac injury, and subsequent reperfusion further enhances the activation of innate and adaptive immune responses and cell death programs. Therefore, inflammation and inflammatory cell infiltration are the hallmarks of MI and reperfusion injury. Ischemic cardiac injury activates the innate immune response via toll-like receptors and upregulates chemokine and cytokine expressions in the infarcted heart. The recruitment of inflammatory cells is a dynamic and superbly orchestrated process. Sequential infiltration of the injured myocardium with neutrophils, monocytes and their descendant macrophages, dendritic cells, and lymphocytes contributes to the initiation and resolution of inflammation, infarct healing, angiogenesis, and ventricular remodeling. Both detrimental effects and a beneficial role in the pathophysiology of MI and reperfusion injury may be attributed to the subset heterogeneity and functional diversity of these inflammatory cells. PMID:27279755

  18. Somatic cell mutations caused by 365 nm LED-UVA due to DNA double-strand breaks through oxidative damage.

    PubMed

    Fang, Xing; Ide, Naohiro; Higashi, Sho-Ichi; Kamei, Yasuhiro; Toyooka, Tatsushi; Ibuki, Yuko; Kawai, Kazuaki; Kasai, Hiroshi; Okamoto, Keinosuke; Arimoto-Kobayashi, Sakae; Negishi, Tomoe

    2014-09-01

    Evidence is accumulating indicating that UVA (320-400 nm ultraviolet light) plays an important role in photo-carcinogenesis. UVA is thought to produce reactive oxygen species in irradiated cells through photo-activation of inherent photosensitizers, and was recently reported to cause DNA double-strand breaks (DSBs) in exposed cells. We have investigated the involvement of UVA in mutations and DNA damage in somatic cells using Drosophila melanogaster larvae. Using the Okazaki Large Spectrograph, we previously observed that longer wavelength UVA (>330 nm) was more mutagenic in post-replication repair-deficient D. melanogaster (mei-41) than in the nucleotide excision repair-deficient strain (mei-9). LED-light has recently been developed as a high-dose-rate UVA source. LED-UVA light (365 nm) was also more mutagenic in mei-41 than in mei-9. The mei-41 gene was shown to be an orthologue of the human ATR gene, which is involved in the repair of DSBs through phosphorylation of histone H2AX. In order to estimate the extent to which oxidative damage contributes to mutation, we established a new D. melanogaster strain (urate-null mutant) that is sensitive to oxidative damage and has a marker to detect somatic cell mutations. When somatic cell mutations were examined using this strain, LED-UVA was mutagenic in the urate-null strain at doses that were non-mutagenic in the urate-positive strain. In an effort to investigate the generation of DSBs, we examined the presence of phosphorylated histone H2AvD (H2AX D. melanogaster homologue). At high doses of LED-UVA (>800 kJ m(-2)), levels of phosphorylated H2AvD (γ-H2AvD) increased significantly in the urate-null strain. Moreover, the level of γ-H2AvD increased in the excision repair-deficient strain but not in the ATR-deficient strain following UVA-irradiation. These results supported the notion that the generation of γ-H2AvD was mediated by the function of the mei-41 gene. It was reported that ATR functions on DSB repair in D

  19. Layered double hydroxide nanoparticles promote self-renewal of mouse embryonic stem cells through the PI3K signaling pathway

    NASA Astrophysics Data System (ADS)

    Wu, Youjun; Zhu, Rongrong; Zhou, Yang; Zhang, Jun; Wang, Wenrui; Sun, Xiaoyu; Wu, Xianzheng; Cheng, Liming; Zhang, Jing; Wang, Shilong

    2015-06-01

    Embryonic stem cells (ESCs) hold great potential for regenerative medicine due to their two unique characteristics: self-renewal and pluripotency. Several groups of nanoparticles have shown promising applications in directing the stem cell fate. Herein, we investigated the cellular effects of layered double hydroxide nanoparticles (LDH NPs) on mouse ESCs (mESCs) and the associated molecular mechanisms. Mg-Al-LDH NPs with an average diameter of ~100 nm were prepared by hydrothermal methods. To determine the influences of LDH NPs on mESCs, cellular cytotoxicity, self-renewal, differentiation potential, and the possible signaling pathways were explored. Evaluation of cell viability, lactate dehydrogenase release, ROS generation and apoptosis demonstrated the low cytotoxicity of LDH NPs. The alkaline phosphatase activity and the expression of pluripotency genes in mESCs were examined, which indicated that exposure to LDH NPs could support self-renewal and inhibit spontaneous differentiation of mESCs under feeder-free culture conditions. The self-renewal promotion was further proved to be independent of the leukemia inhibitory factor (LIF). Furthermore, cells treated with LDH NPs maintained the potential to differentiate into all three germ layers both in vitro and in vivo through formation of embryoid bodies and teratomas. In addition, we observed that LDH NPs initiated the activation of the PI3K/Akt pathway, while treatment with the PI3K inhibitor LY294002 could block the effects of LDH NPs on mESCs. The results confirmed that the promotion of self-renewal by LDH NPs was associated with activation of the PI3K/Akt signaling pathway. Altogether, our studies identified a new role of LDH NPs in maintaining self-renewal of mouse ES cells which could potentially be applied in stem cell research.Embryonic stem cells (ESCs) hold great potential for regenerative medicine due to their two unique characteristics: self-renewal and pluripotency. Several groups of nanoparticles

  20. Antiproliferative Activity of Double Point Modified Analogs of 1,25-Dihydroxyvitamin D2 Against Human Malignant Melanoma Cell Lines

    PubMed Central

    Piotrowska, Anna; Wierzbicka, Justyna; Nadkarni, Sharmin; Brown, Geoffrey; Kutner, Andrzej; Żmijewski, Michał A.

    2016-01-01

    Vitamin D is a lipid soluble steroid hormone with pleiotropic biological properties, including regulation of cell proliferation, differentiation and apoptosis. As to these desirable anticancer actions, 1,25-dihydroxyvitamins D and analogs have been reported to inhibit the proliferation and to induce differentiation of a wide variety of cancer cell types, including human malignant melanoma. However, there is a need for novel and more efficacious vitamin D analogs, and how best to design such is still an open issue. A series of double point modified (DPM) analogs of 1,25-dihydroxyvitamin D2 (1,25(OH)2D2) induced differentiation of the vitamin D receptor (VDR) positive A375 and VDR negative SK-MEL 188b human malignant melanoma cell lines. Surprisingly, the dose of 1,25(OH)2D2 required to inhibit the proliferation of the A375 melanoma cell line by was several fold lower than that required in the case of 1,25(OH)2D3. To evaluate the impact of the modification in the side chain (additional 22-hydroxyl) and in the A-ring (5,6-trans modification), the regular side-chain of vitamin D2 or D3 was retained in the structure of our analogs. As expected, 5,6-trans modification was advantageous to enhancing the anti-proliferative activity of analogs, but not as a single point modification (SPM). Very unexpectedly, the additional 22-hydroxyl in the side-chain reduced significantly the anti-proliferative activity of both the natural and 5,6-trans series analogs. Finally, an induction of pigmentation in melanoma SK-MEL 188b cells was observed to sensitized cells to the effect of vitamin D analogs. PMID:26760999

  1. Transcription-associated processes cause DNA double-strand breaks and translocations in neural stem/progenitor cells

    PubMed Central

    Schwer, Bjoern; Wei, Pei-Chi; Chang, Amelia N.; Kao, Jennifer; Du, Zhou; Meyers, Robin M.; Alt, Frederick W.

    2016-01-01

    High-throughput, genome-wide translocation sequencing (HTGTS) studies of activated B cells have revealed that DNA double-strand breaks (DSBs) capable of translocating to defined bait DSBs are enriched around the transcription start sites (TSSs) of active genes. We used the HTGTS approach to investigate whether a similar phenomenon occurs in primary neural stem/progenitor cells (NSPCs). We report that breakpoint junctions indeed are enriched around TSSs that were determined to be active by global run-on sequencing analyses of NSPCs. Comparative analyses of transcription profiles in NSPCs and B cells revealed that the great majority of TSS-proximal junctions occurred in genes commonly expressed in both cell types, possibly because this common set has higher transcription levels on average than genes transcribed in only one or the other cell type. In the latter context, among all actively transcribed genes containing translocation junctions in NSPCs, those with junctions located within 2 kb of the TSS show a significantly higher transcription rate on average than genes with junctions in the gene body located at distances greater than 2 kb from the TSS. Finally, analysis of repair junction signatures of TSS-associated translocations in wild-type versus classical nonhomologous end-joining (C-NHEJ)–deficient NSPCs reveals that both C-NHEJ and alternative end-joining pathways can generate translocations by joining TSS-proximal DSBs to DSBs on other chromosomes. Our studies show that the generation of transcription-associated DSBs is conserved across divergent cell types. PMID:26873106

  2. Dendritic Cells: A Double-Edged Sword in Immune Responses during Chagas Disease.

    PubMed

    Gil-Jaramillo, Natalia; Motta, Flávia N; Favali, Cecília B F; Bastos, Izabela M D; Santana, Jaime M

    2016-01-01

    Dendritic cells (DCs) are the most important member of the antigen presenting cells group due to their ability to recognize antigen at the infection site and their high specialized antigen internalization capacity. These cells have central role in connecting the innate and adaptive immune responses against Trypanosoma cruzi, the causative agent of Chagas disease. These first line defense cells modulate host immune response depending on type, maturation level, cytokine milieu and DC receptor involved in the interactions with T. cruzi, influencing the development of the disease clinic forms. Here, we present a review of DCs-T. cruzi interactions both in human and murine models, pointing out the parasite ability to manipulate DCs activity for the purpose of evading innate immune response and assuring its own survival and persistence. PMID:27471496

  3. Dendritic Cells: A Double-Edged Sword in Immune Responses during Chagas Disease

    PubMed Central

    Gil-Jaramillo, Natalia; Motta, Flávia N.; Favali, Cecília B. F.; Bastos, Izabela M. D.; Santana, Jaime M.

    2016-01-01

    Dendritic cells (DCs) are the most important member of the antigen presenting cells group due to their ability to recognize antigen at the infection site and their high specialized antigen internalization capacity. These cells have central role in connecting the innate and adaptive immune responses against Trypanosoma cruzi, the causative agent of Chagas disease. These first line defense cells modulate host immune response depending on type, maturation level, cytokine milieu and DC receptor involved in the interactions with T. cruzi, influencing the development of the disease clinic forms. Here, we present a review of DCs–T. cruzi interactions both in human and murine models, pointing out the parasite ability to manipulate DCs activity for the purpose of evading innate immune response and assuring its own survival and persistence. PMID:27471496

  4. On the use of double FLP recognition targets (FRTs) in the LTR of retroviruses for the construction of high producer cell lines.

    PubMed Central

    Karreman, S; Hauser, H; Karreman, C

    1996-01-01

    A pilot experiment for the construction of a hamster derived, high producer cell line using site specific recombination is described. In the experiment chromosomal loci with intrinsic high expression characteristics were sought via infection with a retroviral construct, containing double FRT sites and subsequent screening for overproduction of an encoded markergene. These sites were then targeted with a second vector, that recombined via the FLP/FRT system from Saccharomyces cerevisiae yielding cells that had the second construct at exactly the same position as the first. By using retroviral vectors with double and single FRT sites, respectively, stable clones can be created that can no longer be excised with FLP. PMID:8649977

  5. High efficiency and high photo-stability zinc-phthalocyanine based planar heterojunction solar cells with a double interfacial layer

    NASA Astrophysics Data System (ADS)

    Kim, Tae-Min; Whan Kim, Ji; Shim, Hyun-Sub; Kim, Jang-Joo

    2012-09-01

    The use of CuI and MoO3 as a double interfacial layer between indium tin oxide (ITO) and a zinc phthalocyanine (ZnPc) layer improves the power conversion efficiency (ηp) and the photo-stability at the same time in ZnPc based solar cells. Insertion of CuI without MoO3 increased ηp more than 2 times to 3.3%. However, the photo-stability is lowered even further due to diffusion of Cu. Insertion of the MoO3 layer between the ITO and CuI prevents the diffusion of Cu under UV illumination to achieve the improved photo-stability and ηp.

  6. Intra-Species Bacterial Quorum Sensing Studied at Single Cell Level in a Double Droplet Trapping System

    PubMed Central

    Bai, Yunpeng; Patil, Santoshkumar N.; Bowden, Steven D.; Poulter, Simon; Pan, Jie; Salmond, George P. C.; Welch, Martin; Huck, Wilhelm T. S.; Abell, Chris

    2013-01-01

    In this paper, we investigated the intra-species bacterial quorum sensing at the single cell level using a double droplet trapping system. Escherichia coli transformed to express the quorum sensing receptor protein, LasR, were encapsulated in microdroplets that were positioned adjacent to microdroplets containing the autoinducer, N-(3-oxododecanoyl)- l-homoserine lactone (OdDHL). Functional activation of the LasR protein by diffusion of the OdDHL across the droplet interface was measured by monitoring the expression of green fluorescent protein (GFP) from a LasR-dependent promoter. A threshold concentration of OdDHL was found to induce production of quorum-sensing associated GFP by E. coli. Additionally, we demonstrated that LasR-dependent activation of GFP expression was also initiated when the adjacent droplets contained single E. coli transformed with the OdDHL synthase gene, LasI, representing a simple quorum sensing circuit between two droplets. PMID:23698779

  7. Targeting neoplastic B cells and harnessing microenvironment: the "double face" of ibrutinib and idelalisib.

    PubMed

    Maffei, Rossana; Fiorcari, Stefania; Martinelli, Silvia; Potenza, Leonardo; Luppi, Mario; Marasca, Roberto

    2015-01-01

    Tyrosine kinase inhibitors (TKIs) targeting signaling molecules downstream B cell receptor (BCR) are powerfully spreading in the therapeutic landscape of B cell lymphoproliferative disease, due to a manageable toxicity profile and encouraging clinical effectiveness. In particular, ibrutinib, previously called PCI-32765, is a potent inhibitor of Bruton tyrosine kinase (Btk), recently approved for the treatment of relapsed mantle cell lymphoma (MCL) and chronic lymphocytic leukemia (CLL). Moreover, idelalisib (formerly GS-1101 and CAL-101) is a selective reversible inhibitor of the p110δ isoform of phosphoinositol 3 kinase (PI3K) approved for the treatment of patients with relapsed follicular lymphoma (FL) and CLL. These agents directly affect the neoplastic clone, disrupting the supportive platform provided by BCR signaling cascade and by other microenvironmental mutualistic interactions, and also interfering with chemokine gradients and adhesive properties of neoplastic B cells. In the present review, we describe the clinical efficacy of ibrutinib and idelalisib in CLL and B cell non-Hodgkin lymphoma (B-NHL), then focusing on the mode of action (MOA) of these TKIs towards the neoplastic B cell compartment. At last, the review would further expand the view on potential additional targets of ibrutinib and idelalisib belonging to other microenvironmental cellular elements. PMID:26022368

  8. Penetration of Enveloped Double-Stranded RNA Bacteriophages φ13 and φ6 into Pseudomonas syringae Cells

    PubMed Central

    Daugelavičius, Rimantas; Cvirkaitė, Virginija; Gaidelytė, Aušra; Bakienė, Elena; Gabrėnaitė-Verkhovskaya, Rasa; Bamford, Dennis H.

    2005-01-01

    Bacteriophages φ6 and φ13 are related enveloped double-stranded RNA viruses that infect gram-negative Pseudomonas syringae cells. φ6 uses a pilus as a receptor, and φ13 attaches to the host lipopolysaccharide. We compared the entry-related events of these two viruses, including receptor binding, envelope fusion, peptidoglycan penetration, and passage through the plasma membrane. The infection-related events are dependent on the multiplicity of infection in the case of φ13 but not with φ6. A temporal increase of host outer membrane permeability to lipophilic ions was observed from 1.5 to 4 min postinfection in both virus infections. This enhanced permeability period coincided with the fast dilution of octadecyl rhodamine B-labeled virus-associated lipid molecules. This result is in agreement with membrane fusion, and the presence of temporal virus-derived membrane patches on the outer membrane. Similar to φ6, φ13 contains a thermosensitive lytic enzyme involved in peptidoglycan penetration. The phage entry also caused a limited depolarization of the plasma membrane. Inhibition of host respiration considerably decreased the efficiency of irreversible virus binding and membrane fusion. An active role of cell energy metabolism in restoring the infection-induced defects in the cell envelope was also observed. PMID:15795287

  9. Unrepairable DNA double-strand breaks that are generated by ionising radiation determine the fate of normal human cells.

    PubMed

    Noda, Asao; Hirai, Yuko; Hamasaki, Kanya; Mitani, Hiroshi; Nakamura, Nori; Kodama, Yoshiaki

    2012-11-15

    After an exposure to ionising radiation, cells can quickly repair damage to their genomes; however, a few unrepairable DNA double-strand breaks (DSBs) emerge in the nucleus in a prolonged culture and perpetuate as long as the culture continues. These DSBs may be retained forever in cells such as non-dividing ageing tissues, which are resistant to apoptosis. We show that such unrepairable DSBs, which had been advocated by the classical target theory as the 'radiation hit', could account for permanent growth arrest and premature senescence. The unrepairable DSBs build up with repeated irradiation, which accounts for an accumulated dose. Because these DSBs tend to be paired, we propose that the untethered and 'torn-off' molecular structures at the broken ends of the DNA result in an alteration of chromatin structure, which protects the ends of the DNA from genomic catastrophe. Such biochemical responses are important for cell survival but may cause gradual tissue malfunction, which could lead to the late effects of radiation exposure. Thus, understanding the biology of unrepairable damage will provide new insights into the long-term effects of radiation. PMID:22899723

  10. One-step generation of cell-laden microgels using double emulsion drops with a sacrificial ultra-thin oil shell.

    PubMed

    Choi, Chang-Hyung; Wang, Huanan; Lee, Hyomin; Kim, June Hwan; Zhang, Liyuan; Mao, Angelo; Mooney, David J; Weitz, David A

    2016-04-26

    Cell-laden microgels with highly uniform sizes have significant potential in tissue engineering and cell therapy due to their capability to provide a physiologically relevant three-dimensional (3D) microenvironment for living cells. In this work, we present a simple and efficient microfluidic approach to produce monodisperse cell-laden microgels through the use of double emulsion drops with an ultra-thin oil shell as the sacrificial template. Specifically, the thin oil shell in double emulsion spontaneously dewets upon polymerization of the innermost precursor drop and subsequent transfer into an aqueous solution, resulting in direct dispersion of microgels in the aqueous phase. Compared to conventional single emulsion-based techniques for cell encapsulation, this one-step approach prevents prolonged exposure of cells to the oil phase, leading to high-throughput cell encapsulation in microgels without compromising the cell viability. Moreover, this approach allows us to culture cells within a 3D microgel which mimics the extracellular matrix, thus enabling long-term cell functionality. This microfluidic technique represents a significant step forward in high-throughput cell microencapsulation technology and offers a potentially viable option to produce cell-laden microgels for widespread applications in tissue engineering and cell therapies. PMID:27070224

  11. A hybrid microbial fuel cell stack based on single and double chamber microbial fuel cells for self-sustaining pH control

    NASA Astrophysics Data System (ADS)

    Yang, Wei; Li, Jun; Ye, Dingding; Zhang, Liang; Zhu, Xun; Liao, Qiang

    2016-02-01

    Proton accumulation in the anode chamber is the major problem that affects the operational stability and electricity generation performance of double chamber microbial fuel cells (MFCs). In this study, a hybrid microbial fuel cell stack (DS-DS stack) based on single (SCMFCs) and double chamber MFCs (DCMFCs) is proposed for self-sustaining pH control in the MFC stack. It is found that the aerobic microbial oxidation of acetate by the biofilm that is attached to the air cathode of SCMFCs is responsible for the self-sustaining removal of accumulated H+ in the effluent of DCMFCs. Compared with the stack that solely consists of SCMFCs (SS-SS stack) or DCMFCs (DD-DD stack), the hybrid stack exhibits the highest electricity output performance and the most effective conversion of acetate into electricity at high power levels. Furthermore, the hybrid stack demonstrates the operation time of 15.7 ± 1.1 h when the operating voltage is above 0.8 V. This value is much higher than that of the DD-DD (8.5 ± 2.4 h) and SS-SS (8.1 ± 1.4 h) stacks, which suggests that the hybrid stack had a good operational stability.

  12. Long-term operation of double chambered microbial fuel cell for bio-electro denitrification.

    PubMed

    Oon, Yoong-Sin; Ong, Soon-An; Ho, Li-Ngee; Wong, Yee-Shian; Oon, Yoong-Ling; Lehl, Harvinder Kaur; Thung, Wei-Eng

    2016-06-01

    The main aim of this study is to investigate the performance of organic oxidation and denitrification of the system under long-term operation. The MFC reactor was operated in continuous mode for 180 days. Nitrate was successfully demonstrated as terminal electron acceptor, where nitrate was reduced at the cathode using electron provided by acetate oxidation at the anode. The removal efficiencies of chemical oxygen demand (COD) and nitrate were higher in the closed circuit system than in open circuit system. Both COD and nitrate reduction improved with the increase of organic loading and subsequently contributed to higher power output. The maximum nitrate removal efficiency was 88 ± 4 % (influent of 141 ± 14 mg/L). The internal resistant was 50 Ω, which was found to be low for a double chambered MFC. The maximum power density was 669 mW/m(3) with current density of 3487 mA/m(3). PMID:26894384

  13. Assembly and function of DNA double-strand break repair foci in mammalian cells.

    PubMed

    Bekker-Jensen, Simon; Mailand, Niels

    2010-12-10

    DNA double-strand breaks (DSBs) are among the most cytotoxic types of DNA damage, which if left unrepaired can lead to mutations or gross chromosomal aberrations, and promote the onset of diseases associated with genomic instability such as cancer. One of the most discernible hallmarks of the cellular response to DSBs is the accumulation and local concentration of a plethora of DNA damage signaling and repair proteins in the vicinity of the lesion, initiated by ATM-mediated phosphorylation of H2AX (γ-H2AX) and culminating in the generation of distinct nuclear compartments, so-called Ionizing Radiation-Induced Foci (IRIF). The assembly of proteins at the DSB-flanking chromatin occurs in a highly ordered and strictly hierarchical fashion. To a large extent, this is achieved by regulation of protein-protein interactions triggered by a variety of post-translational modifications including phosphorylation, ubiquitylation, SUMOylation, and acetylation. Over the last decade, insight into the identity of proteins residing in IRIF and the molecular underpinnings of their retention at these structures has been vastly expanded. Despite such advances, however, our understanding of the biological relevance of such DNA repair foci still remains limited. In this review, we focus on recent discoveries on the mechanisms that govern the formation of IRIF, and discuss the implications of such findings in light of our understanding of the physiological importance of these structures. PMID:21035408

  14. Identification of defective illegitimate recombinational repair of oxidatively-induced DNA double-strand breaks in ataxia-telangiectasia cells

    NASA Technical Reports Server (NTRS)

    Dar, M. E.; Winters, T. A.; Jorgensen, T. J.

    1997-01-01

    Ataxia-telangiectasia (A-T) is an autosomal-recessive lethal human disease. Homozygotes suffer from a number of neurological disorders, as well as very high cancer incidence. Heterozygotes may also have a higher than normal risk of cancer, particularly for the breast. The gene responsible for the disease (ATM) has been cloned, but its role in mechanisms of the disease remain unknown. Cellular A-T phenotypes, such as radiosensitivity and genomic instability, suggest that a deficiency in the repair of DNA double-strand breaks (DSBs) may be the primary defect; however, overall levels of DSB rejoining appear normal. We used the shuttle vector, pZ189, containing an oxidatively-induced DSB, to compare the integrity of DSB rejoining in one normal and two A-T fibroblast cells lines. Mutation frequencies were two-fold higher in A-T cells, and the mutational spectrum was different. The majority of the mutations found in all three cell lines were deletions (44-63%). The DNA sequence analysis indicated that 17 of the 17 plasmids with deletion mutations in normal cells occurred between short direct-repeat sequences (removing one of the repeats plus the intervening sequences), implicating illegitimate recombination in DSB rejoining. The combined data from both A-T cell lines showed that 21 of 24 deletions did not involve direct-repeats sequences, implicating a defect in the illegitimate recombination pathway. These findings suggest that the A-T gene product may either directly participate in illegitimate recombination or modulate the pathway. Regardless, this defect is likely to be important to a mechanistic understanding of this lethal disease.

  15. Double-layer composite film based on hollow TiO{sub 2} boxes and P25 as photoanode for enhanced efficiency in dye-sensitized solar cells

    SciTech Connect

    Shi, Yue; Zhao, Li Wang, Shimin Li, Jing; Dong, Binghai; Xu, Zuxun; Wan, Li

    2014-11-15

    Highlights: • A novel TiO{sub 2} double-layer composite film is successfully fabricated. • The TiO{sub 2}-HB overlayer exhibited intense scattering ability. • The P25 particles underlayer can enlarge the contact area and increase the contact points between TiO{sub 2}-HB and FTO glass. • TiO{sub 2}-DLL film cells have the maximum conversion efficiency. - Abstract: A TiO{sub 2} double-layer composite film consisting of hollow TiO{sub 2} boxes (HB-TiO{sub 2}) as overlayer and commercial-grade TiO{sub 2} nanoparticles (P25) as underlayer is designed as the photoelectrode of dye-sensitized solar cells (DSSCs). The hollow TiO{sub 2} box has been employed as light-scattering part to increase the optical length in the film and enhance light-harvesting. It was found that the double-layer film cell with lower absorbed dye achieved the highest conversion efficiency and reach up to a maximum value of 6.4%, which is 226% higher than that of pure HB-TiO{sub 2} film cell (η = 1.96%) and 57.2% higher than that of pure P25 particles film cell under identical film thickness at a constant irradiation of 100 mW cm{sup −2}. The enhanced efficiency of double-layer film can be attributed to its light-scattering capability.

  16. CIGS absorber layer with double grading Ga profile for highly efficient solar cells

    NASA Astrophysics Data System (ADS)

    Saadat, M.; Moradi, M.; Zahedifar, M.

    2016-04-01

    It is well-known that the band gap grading in CIGS solar cells is crucial for achieving highly efficient solar cells. We stimulate a CIGS solar cell and investigate the effects of the band gap grading on performance of the CIGS solar cell, where Ga/(Ga + In) ratio (GGI) at back (Cb) and front (Cf) of the absorber layer are considered constant. Our simulations show that by increasing the GGI at middle of CIGS absorber layer (Cm), the JSC decreases and VOC increases independent of the distance of the Cm from the back contact (Xm). For Cm lower than Cf, JSC increases and VOC decreases when the Xm shifts to the front of the CIGS layer. The behavior of JSC and VOC became reverse for the case of Cm greater than Cf. Almost in all of the structures, efficiency and FF have same behaviors. Our simulations show that the highest efficiency is obtained at Cm = 0.8 and Xm = 200 nm.

  17. Mechanisms of double-strand-break repair during gene targeting in mammalian cells.

    PubMed Central

    Ng, P; Baker, M D

    1999-01-01

    In the present study, the mechanism of double-strand-break (DSB) repair during gene targeting at the chromosomal immunoglobulin mu-locus in a murine hybridoma was examined. The gene-targeting assay utilized specially designed insertion vectors genetically marked in the region of homology to the chromosomal mu-locus by six diagnostic restriction enzyme site markers. The restriction enzyme markers permitted the contribution of vector-borne and chromosomal mu-sequences in the recombinant product to be determined. The use of the insertion vectors in conjunction with a plating procedure in which individual integrative homologous recombination events were retained for analysis revealed several important features about the mammalian DSB repair process:The presence of the markers within the region of shared homology did not affect the efficiency of gene targeting.In the majority of recombinants, the vector-borne marker proximal to the DSB was absent, being replaced with the corresponding chromosomal restriction enzyme site. This result is consistent with either formation and repair of a vector-borne gap or an "end" bias in mismatch repair of heteroduplex DNA (hDNA) that favored the chromosomal sequence. Formation of hDNA was frequently associated with gene targeting and, in most cases, began approximately 645 bp from the DSB and could encompass a distance of at least 1469 bp.The hDNA was efficiently repaired prior to DNA replication.The repair of adjacent mismatches in hDNA occurred predominantly on the same strand, suggesting the involvement of a long-patch repair mechanism. PMID:10049929

  18. Scaling of volume to surface ratio and doubling time in growing unicellular organisms: Do cells appear quantum-mechanical systems?

    NASA Astrophysics Data System (ADS)

    Atanasov, Atanas Todorov

    2014-10-01

    The scaling of physical and biological characteristics of the living organisms is a basic method for searching of new biophysical laws. In series of previous studies the author showed that in Poikilotherms, Mammals and Aves, the volume to surface ratio V×S-1 (m) of organisms is proportional to their generation time Tgt(s) via growth rate v (m s-1): V×S-1 = vgr×Tr. The power and the correlation coefficients are near to 1.0. Aim of this study is: i) to prove with experimental data the validity of the above equation for Unicellular organisms and ii) to show that perhaps, the cells are quantum-mechanical systems. The data for body mass M (kg), density ρ (kg/m3), minimum and maximum doubling time Tdt (s) for 50 unicellular organisms are assembled from scientific sources, and the computer program `Statistics' is used for calculations. In result i) the analytical relationship from type: V×S-1 = 4.46ṡ10-11×Tdt was found, where vgr = 4.46×10-11 m/s and ii) it is shown that the products between cell mass M, cell length expressed by V/S ratio and growth rate vgr satisfied the Heisenberg uncertainty principle i.e. the inequalities V/S×M×vgr>h/2π and Tdt×M×vgr2>h/2π are valid, where h= 6.626×10-34 Jṡs is the Planck constant. This rise the question: do cells appear quantum-mechanical systems?

  19. Scaling of volume to surface ratio and doubling time in growing unicellular organisms: Do cells appear quantum-mechanical systems?

    SciTech Connect

    Atanasov, Atanas Todorov

    2014-10-06

    The scaling of physical and biological characteristics of the living organisms is a basic method for searching of new biophysical laws. In series of previous studies the author showed that in Poikilotherms, Mammals and Aves, the volume to surface ratio V×S{sup −1} (m) of organisms is proportional to their generation time T{sub gt}(s) via growth rate v (m s{sup −1}): V×S{sup −1} = v{sub gr}×T{sup r}. The power and the correlation coefficients are near to 1.0. Aim of this study is: i) to prove with experimental data the validity of the above equation for Unicellular organisms and ii) to show that perhaps, the cells are quantum-mechanical systems. The data for body mass M (kg), density ρ (kg/m{sup 3}), minimum and maximum doubling time T{sub dt} (s) for 50 unicellular organisms are assembled from scientific sources, and the computer program ‘Statistics’ is used for calculations. In result i) the analytical relationship from type: V×S{sup −1} = 4.46⋅10{sup −11}×T{sub dt} was found, where v{sub gr} = 4.46×10{sup −11} m/s and ii) it is shown that the products between cell mass M, cell length expressed by V/S ratio and growth rate v{sub gr} satisfied the Heisenberg uncertainty principle i.e. the inequalities V/S×M×v{sub gr}>h/2π and T{sub dt}×M×v{sub gr}{sup 2}>h/2π are valid, where h= 6.626×10{sup −34} J⋅s is the Planck constant. This rise the question: do cells appear quantum-mechanical systems?.

  20. Multistrain Probiotic Modulation of Intestinal Epithelial Cells' Immune Response to a Double-Stranded RNA Ligand, Poly(I·C)

    PubMed Central

    MacPherson, Chad; Audy, Julie; Mathieu, Olivier

    2014-01-01

    A commercially available product containing three probiotic bacterial strains (Lactobacillus helveticus R0052, Bifidobacterium longum subsp. infantis R0033, and Bifidobacterium bifidum R0071) was previously shown in animal trials to modulate both TH1 and TH2 immune responses. Clinical studies on this combination of bacteria have also shown positive health effects against seasonal winter diseases and rotavirus infection. The goal of this study was to use a well-established in vitro intestinal epithelial (HT-29) cell model that has been shown to constitutively express double-stranded RNA (dsRNA) sensors (Toll-like receptor 3 [TLR3], retinoic acid-inducible gene I, melanoma differentiation-associated gene 5, and dsRNA-activated protein kinase). By using the HT-29 cell model, we wanted to evaluate whether or not this combination of three bacteria had the capacity to immune modulate the host cell response to a dsRNA ligand, poly(I·C). Using a custom-designed, two-color expression microarray targeting genes of the human immune system, we investigated the response of HT-29 cells challenged with poly(I·C) both in the presence and in the absence of the three probiotic bacteria. We observed that the combination of the three bacteria had a major impact on attenuating the expression of genes connected to proinflammatory TH1 and antiviral innate immune responses compared to that obtained by the poly(I·C)-only challenge. Major pathways through which the multistrain combination may be eliciting its immune-modulatory effect include the TLR3 domain-containing adapter-inducing beta interferon (TRIF), mitogen-activated protein kinase, and NF-κB signaling pathways. Such a model may be useful for selecting potential biomarkers for the design of future clinical trials. PMID:24375132

  1. HDAC inhibition radiosensitizes human normal tissue cells and reduces DNA Double-Strand Break repair capacity.

    PubMed

    Purrucker, Jan C; Fricke, Andreas; Ong, Mei Fang; Rübe, Christian; Rübe, Claudia E; Mahlknecht, Ulrich

    2010-01-01

    HDAC inhibitors (HDACi) are gaining increasing attention in the treatment of cancer, particularly in view of their therapeutic effectiveness and assumed mild toxicity profile. While numerous studies have investigated the role of HDACi in tumor cells, little is known about their effects on normal tissue cells. We studied the effect of suberoylanilide hydroxamic acid (SAHA), MS275, sodium-butyrate and valproic acid in healthy human fibroblasts and found HDACi-treatment to go along with increased radiosensitivity and reduced DSB repair capacity. In view of the potential genotoxic effects of HDACi-treatment, particularly when being administered long-term for chronic disease or when given to children, to women of childbearing age or their partners or in combination with radiotherapy, an extensive education of patients and prescribing physicians as well as a stringent definition of clinical indications is urgently required. PMID:19956891

  2. Investigation of the double exponential in the current-voltage characteristics of silicon solar cells

    NASA Technical Reports Server (NTRS)

    Wolf, M.; Noel, G. T.; Stirn, R. J.

    1976-01-01

    A theoretical analysis is presented of certain peculiarities of the current-voltage characteristics of silicon solar cells, involving high values of the empirical constant A in the diode equation for a p-n junction. An attempt was made in a lab experiment to demonstrate that the saturation current which is associated with the exponential term qV/A2kT of the I-V characteristic, with A2 roughly equal to 2, originates in the space charge region and that it can be increased, as observed on ATS-1 cells, by the introduction of additional defects through low energy proton irradiation. It was shown that the proton irradiation introduces defects into the space charge region which give rise to a recombination current from this region, although the I-V characteristic is, in this case, dominated by an exponential term which has A = 1.

  3. The role of buffer layers and double windows layers in a solar cell CZTS performances

    NASA Astrophysics Data System (ADS)

    Mebarkia, C.; Dib, D.; Zerfaoui, H.; Belghit, R.

    2016-07-01

    In the overall context of the diversification of the use of natural resources, the use of renewable energy including solar photovoltaic has become increasingly indispensable. As such, the development of a new generation of photovoltaic cells based on CuZnSnS4 (CZTS) looks promising. Cu2ZnSnS4 (CZTS) is a new film absorber, with good physical properties (band gap energy 1.4-1.6 eV with a large absorption coefficient over 104 cm-1). Indeed, the performance of these cells exceeded 30% in recent years. In the present paper, our work based on modeling and numerical simulation, we used SCAPS to study the performance of solar cells based on Cu2ZnSnS4 (CZTS) and thus evaluate the electrical efficiency η for typical structures of n-ZnO:Al / i-ZnO / n-CdS / p-CZTS and n-ITO / n-ZnO:Al / n-CdS /p-CZTS. Furthermore, the influence of the change of CdS by ZnSeand In2S3buffer layer was treated in this paper.

  4. Leishmania chagasi T-cell antigens identified through a double library screen.

    PubMed

    Martins, Daniella R A; Jeronimo, Selma M B; Donelson, John E; Wilson, Mary E

    2006-12-01

    Control of human visceral leishmaniasis in regions where it is endemic is hampered in part by limited accessibility to medical care and emerging drug resistance. There is no available protective vaccine. Leishmania spp. protozoa express multiple antigens recognized by the vertebrate immune system. Since there is not one immunodominant epitope recognized by most hosts, strategies must be developed to optimize selection of antigens for prevention and immunodiagnosis. For this reason, we generated a cDNA library from the intracellular amastigote form of Leishmania chagasi, the cause of South American visceral leishmaniasis. We employed a two-step expression screen of the library to systematically identify T-cell antigens and T-dependent B-cell antigens. The first step was aimed at identifying the largest possible number of clones producing an epitope-containing polypeptide by screening with a pool of sera from Brazilians with documented visceral leishmaniasis. After removal of clones encoding heat shock proteins, positive clones underwent a second-step screen for their ability to cause proliferation and gamma interferon responses in T cells from immune mice. Six unique clones were selected from the second screen for further analysis. The corresponding antigens were derived from glutamine synthetase, a transitional endoplasmic reticulum ATPase, elongation factor 1gamma, kinesin K39, repetitive protein A2, and a hypothetical conserved protein. Humans naturally infected with L. chagasi mounted both cellular and antibody responses to these proteins. Preparations containing multiple antigens may be optimal for immunodiagnosis and protective vaccines. PMID:17000724

  5. Polymer Acceptor Based on Double B←N Bridged Bipyridine (BNBP) Unit for High-Efficiency All-Polymer Solar Cells.

    PubMed

    Long, Xiaojing; Ding, Zicheng; Dou, Chuandong; Zhang, Jidong; Liu, Jun; Wang, Lixiang

    2016-08-01

    A novel polymer acceptor based on the double B←N bridged bipyridine building block is reported. All-polymer solar cells based on the new polymer acceptor show a power conversion efficiency of as high as 6.26% at a photon energy loss of only 0.51 eV. PMID:27167123

  6. Layered oxygen-deficient double perovskite as an efficient and stable anode for direct hydrocarbon solid oxide fuel cells.

    PubMed

    Sengodan, Sivaprakash; Choi, Sihyuk; Jun, Areum; Shin, Tae Ho; Ju, Young-Wan; Jeong, Hu Young; Shin, Jeeyoung; Irvine, John T S; Kim, Guntae

    2015-02-01

    Different layered perovskite-related oxides are known to exhibit important electronic, magnetic and electrochemical properties. Owing to their excellent mixed-ionic and electronic conductivity and fast oxygen kinetics, cation layered double perovskite oxides such as PrBaCo2O5 in particular have exhibited excellent properties as solid oxide fuel cell oxygen electrodes. Here, we show for the first time that related layered materials can be used as high-performance fuel electrodes. Good redox stability with tolerance to coking and sulphur contamination from hydrocarbon fuels is demonstrated for the layered perovskite anode PrBaMn2O5+δ (PBMO). The PBMO anode is fabricated by in situ annealing of Pr0.5Ba0.5MnO3-δ in fuel conditions and actual fuel cell operation is demonstrated. At 800 °C, layered PBMO shows high electrical conductivity of 8.16 S cm(-1) in 5% H2 and demonstrates peak power densities of 1.7 and 1.3 W cm(-2) at 850 °C using humidified hydrogen and propane fuels, respectively. PMID:25532072

  7. Visualizing double-stranded RNA distribution and dynamics in living cells by dsRNA binding-dependent fluorescence complementation.

    PubMed

    Cheng, Xiaofei; Deng, Ping; Cui, Hongguang; Wang, Aiming

    2015-11-01

    Double-stranded RNA (dsRNA) is an important type of RNA that plays essential roles in diverse cellular processes in eukaryotic organisms and a hallmark in infections by positive-sense RNA viruses. Currently, no in vivo technology has been developed for visualizing dsRNA in living cells. Here, we report a dsRNA binding-dependent fluorescence complementation (dRBFC) assay that can be used to efficiently monitor dsRNA distribution and dynamics in vivo. The system consists of two dsRNA-binding proteins, which are fused to the N- and C-terminal halves of the yellow fluorescent protein (YFP). Binding of the two fusion proteins to a common dsRNA brings the split YFP halves in close proximity, leading to the reconstitution of the fluorescence-competent structure and restoration of fluorescence. Using this technique, we were able to visualize the distribution and trafficking of the replicative RNA intermediates of positive-sense RNA viruses in living cells. PMID:26351203

  8. Cohesin phosphorylation and mobility of SMC1 at ionizing radiation-induced DNA double-strand breaks in human cells

    SciTech Connect

    Bauerschmidt, Christina; Helleday, Thomas

    2011-02-01

    Cohesin, a hetero-tetrameric complex of SMC1, SMC3, Rad21 and Scc3, associates with chromatin after mitosis and holds sister chromatids together following DNA replication. Following DNA damage, cohesin accumulates at and promotes the repair of DNA double-strand breaks. In addition, phosphorylation of the SMC1/3 subunits contributes to DNA damage-induced cell cycle checkpoint regulation. The aim of this study was to determine the regulation and consequences of SMC1/3 phosphorylation as part of the cohesin complex. We show here that the ATM-dependent phosphorylation of SMC1 and SMC3 is mediated by H2AX, 53BP1 and MDC1. Depletion of RAD21 abolishes these phosphorylations, indicating that only the fully assembled complex is phosphorylated. Comparison of wild type SMC1 and SMC1S966A in fluorescence recovery after photo-bleaching experiments shows that phosphorylation of SMC1 is required for an increased mobility after DNA damage in G2-phase cells, suggesting that ATM-dependent phosphorylation facilitates mobilization of the cohesin complex after DNA damage.

  9. Layered oxygen-deficient double perovskite as an efficient and stable anode for direct hydrocarbon solid oxide fuel cells

    NASA Astrophysics Data System (ADS)

    Sengodan, Sivaprakash; Choi, Sihyuk; Jun, Areum; Shin, Tae Ho; Ju, Young-Wan; Jeong, Hu Young; Shin, Jeeyoung; Irvine, John T. S.; Kim, Guntae

    2015-02-01

    Different layered perovskite-related oxides are known to exhibit important electronic, magnetic and electrochemical properties. Owing to their excellent mixed-ionic and electronic conductivity and fast oxygen kinetics, cation layered double perovskite oxides such as PrBaCo2O5 in particular have exhibited excellent properties as solid oxide fuel cell oxygen electrodes. Here, we show for the first time that related layered materials can be used as high-performance fuel electrodes. Good redox stability with tolerance to coking and sulphur contamination from hydrocarbon fuels is demonstrated for the layered perovskite anode PrBaMn2O5+δ (PBMO). The PBMO anode is fabricated by in situ annealing of Pr0.5Ba0.5MnO3-δ in fuel conditions and actual fuel cell operation is demonstrated. At 800 °C, layered PBMO shows high electrical conductivity of 8.16 S cm-1 in 5% H2 and demonstrates peak power densities of 1.7 and 1.3 W cm-2 at 850 °C using humidified hydrogen and propane fuels, respectively.

  10. 5-Fluorouracil sensitizes colorectal tumor cells towards double stranded DNA breaks by interfering with homologous recombination repair

    PubMed Central

    Srinivas, Upadhyayula Sai; Dyczkowski, Jerzy; Beißbarth, Tim; Gaedcke, Jochen; Mansour, Wael Y.; Borgmann, Kerstin; Dobbelstein, Matthias

    2015-01-01

    Malignant tumors of the rectum are treated by neoadjuvant radiochemotherapy. This involves a combination of 5-fluorouracil (5-FU) and double stranded DNA-break (DSB)-inducing radiotherapy. Here we explored how 5-FU cooperates with DSB-induction to achieve sustainable DNA damage in colorectal cancer (CRC) cells. After DSB induction by neocarzinostatin, phosphorylated histone 2AX (γ-H2AX) rapidly accumulated but then largely vanished within a few hours. In contrast, when CRC cells were pre-treated with 5-FU, gammaH2AX remained for at least 24 hours. GFP-reporter assays revealed that 5-FU decreases the efficiency of homologous recombination (HR) repair. However, 5-FU did not prevent the initial steps of HR repair, such as the accumulation of RPA and Rad51 at nuclear foci. Thus, we propose that 5-FU interferes with the continuation of HR repair, e. g. the synthesis of new DNA strands. Two key mediators of HR, Rad51 and BRCA2, were found upregulated in CRC biopsies as compared to normal mucosa. Inhibition of HR by targeting Rad51 enhanced DNA damage upon DSB-inducing treatment, outlining an alternative way of enhancing therapeutic efficacy. Taken together, our results strongly suggest that interfering with HR represents a key mechanism to enhance the efficacy when treating CRC with DNA-damaging therapy. PMID:25909291

  11. Tetraploidization events by chromosome doubling of nucellar cells are frequent in apomictic citrus and are dependent on genotype and environment

    PubMed Central

    Aleza, Pablo; Froelicher, Yann; Schwarz, Sergio; Agustí, Manuel; Hernández, María; Juárez, José; Luro, François; Morillon, Raphael; Navarro, Luis; Ollitrault, Patrick

    2011-01-01

    Background and Aims Polyploidy is a major component of plant evolution. The citrus gene pool is essentially diploid but tetraploid plants are frequently encountered in seedlings of diploid apomictic genotypes. The main objectives of the present study were to establish the origin of these tetraploid plants and to ascertain the importance of genotypic and environmental factors on tetraploid formation. Methods Tetraploid seedlings from 30 diploid apomictic genotypes were selected by flow cytometry and genotyped with 24 single sequence repeat (SSR) markers to analyse their genetic origin. Embryo rescue was used to grow all embryos contained in polyembryonic seeds of ‘Tardivo di Ciaculli’ mandarin, followed by characterization of the plantlets obtained by flow cytometry and SSR markers to accurately establish the rate of tetraploidization events and their potential tissue location. Inter-annual variations in tetraploid seedling rates were analysed for seven genotypes. Variation in tetraploid plantlet rates was analysed between different seedlings of the same genotype (‘Carrizo’ citrange; Citrus sinensis × Poncirus trifoliata) from seeds collected in different tropical, subtropical and Mediterranean countries. Key Results Tetraploid plants were obtained for all the studied diploid genotypes, except for four mandarins. All tetraploid plants were identical to their diploid maternal line for SSR markers and were not cytochimeric. Significant genotypic and environmental effects were observed, as well as negative correlation between mean temperature during the flowering period and tetraploidy seedling rates. The higher frequencies (20 %) of tetraploids were observed for citranges cultivated in the Mediterranean area. Conclusions Tetraploidization by chromosome doubling of nucellar cells are frequent events in apomictic citrus, and are affected by both genotypic and environmental factors. Colder conditions in marginal climatic areas appear to favour the expression of

  12. Hydrodynamically induced fluid transfer and non-convective double-diffusion in microgravity sliding solvent diffusion cells

    NASA Technical Reports Server (NTRS)

    Pollmann, Konrad W.; Stodieck, Louis S.; Luttges, Marvin W.

    1994-01-01

    Microgravity can provide a diffusion-dominated environment for double-diffusion and diffusion-reaction experiments otherwise disrupted by buoyant convection or sedimentation. In sliding solvent diffusion cells, a diffusion interface between two liquid columns is achieved by aligning two offset sliding wells. Fluid in contact with the sliding lid of the cavities is subjected to an applied shear stress. The momentum change by the start/stop action of the well creates an additional hydrodynamical force. In microgravity, these viscous and inertial forces are sufficiently large to deform the diffusion interface and induce hydrodynamic transfer between the wells. A series of KC-135 parabolic flight experiments were conducted to characterize these effects and establish baseline data for microgravity diffusion experiments. Flow visualizations show the diffusion interface to be deformed in a sinusoidal fashion following well alignment. After the wells were separated again in a second sliding movement, the total induced liquid transfer was determined and normalized by the well aspect ratio. The normalized transfer decreased linearly with Reynolds number from 3.3 to 4.0% (w/v) for Re = 0.4 (Stokes flow) to a minimum of 1.0% for Re = 23 to 30. Reynolds numbers that provide minimum induced transfers are characterized by an interface that is highly deformed and unsuitable for diffusion measurements. Flat diffusion interfaces acceptable for diffusion measurements are obtained with Reynolds numbers on the order of 7 to 10. Microgravity experiments aboard a sounding rocket flight verified counterdiffusion of different solutes to be diffusion dominated. Ground control experiments showed enhanced mixing by double-diffusive convection. Careful selection of experimental parameters improves initial conditions and minimizes induced transfer rates.

  13. MoO3/poly(9,9-dioctylfluorene-co-N-[4-(3-methylpropyl)]-diphenylamine) double-interlayer effect on polymer solar cells

    NASA Astrophysics Data System (ADS)

    Subbiah, Jegadesan; Kim, Do Young; Hartel, Michael; So, Franky

    2010-02-01

    A double interlayer composed of MoO3 and poly(9,9-dioctylfluorene-co-N-[4-(3-methylpropyl)]-diphenylamine) (TFB) was used as an anode contact for bulk heterojunction polymer solar cells. Using this strategy, photovoltaic cells with poly[2-methoxy-5-(3',7'-dimethyloctyloxy)-1,4-phenylene vinylene]: [6,6]-phenyl-C61 butyric acid methyl ester (MDMO-PPV:PCBM) blend as a photoactive layer were fabricated. An enhancement in power conversion efficiency of 53% was observed in cells with a double interlayer compared with cells having a PEDOT: PSS interlayer. The enhancement is attributed to the combined effects of electron blocking and enhanced charge extraction from the photoactive layer to the anode.

  14. High efficiency Cu2ZnSn(S,Se)4 solar cells by applying a double In2S3/CdS emitter.

    PubMed

    Kim, Jeehwan; Hiroi, Homare; Todorov, Teodor K; Gunawan, Oki; Kuwahara, Masaru; Gokmen, Tayfun; Nair, Dhruv; Hopstaken, Marinus; Shin, Byungha; Lee, Yun Seog; Wang, Wei; Sugimoto, Hiroki; Mitzi, David B

    2014-11-26

    High-efficiency Cu2ZnSn(S,Se)4 solar cells are reported by applying In2S3/CdS double emitters. This new structure offers a high doping concentration within the Cu2ZnSn(S,Se)4 solar cells, resulting in a substantial enhancement in open-circuit voltage. The 12.4% device is obtained with a record open-circuit voltage deficit of 593 mV. PMID:25155874

  15. GEM-loaded magnetic albumin nanospheres modified with cetuximab for simultaneous targeting, magnetic resonance imaging, and double-targeted thermochemotherapy of pancreatic cancer cells

    PubMed Central

    Wang, Ling; An, Yanli; Yuan, Chenyan; Zhang, Hao; Liang, Chen; Ding, Fengan; Gao, Qi; Zhang, Dongsheng

    2015-01-01

    Background Targeted delivery is a promising strategy to improve the diagnostic imaging and therapeutic effect of cancers. In this paper, novel cetuximab (C225)-conjugated, gemcitabine (GEM)-containing magnetic albumin nanospheres (C225-GEM/MANs) were fabricated and applied as a theranostic nanocarrier to conduct simultaneous targeting, magnetic resonance imaging (MRI), and double-targeted thermochemotherapy against pancreatic cancer cells. Methods Fe3O4 nanoparticles (NPs) and GEM co-loaded albumin nanospheres (GEM/MANs) were prepared, and then C225 was further conjugated to synthesize C225-GEM/MANs. Their morphology, mean particle size, GEM encapsulation ratio, specific cell-binding ability, and thermal dynamic profiles were characterized. The effects of discriminating different EGFR-expressing pancreatic cancer cells (AsPC-1 and MIA PaCa-2) and monitoring cellular targeting effects were assessed by targeted MRI. Lastly, the antitumor efficiency of double/C225/magnetic-targeted and nontargeted thermochemotherapy was compared with chemotherapy alone using 3-(4, 5-dimethyl-2-thiazolyl)-2,5-diphenyl-2H-tetrazolium bromide (MTT) and flow cytometry (FCM) assay. Results When treated with targeted nanospheres, AsPC-1 cells showed a significantly less intense MRI T2 signal than MIA PaCa-2 cells, while both cells had similar signal strength when incubated with nontargeted nanospheres. T2 signal intensity was significantly lower when magnetic and C225 targeting were combined, rather than used alone. The inhibitory and apoptotic rates of each thermochemotherapy group were significantly higher than those of the chemotherapy-alone groups. Additionally, both MTT and FCM analysis verified that double-targeted thermochemotherapy had the highest targeted killing efficiency among all groups. Conclusion The C225-GEM/MANs can distinguish various EGFR-expressing live pancreatic cancer cells, monitor diverse cellular targeting effects using targeted MRI imaging, and efficiently mediate

  16. Theoretical results on the double-collecting tandem junction solar cell. [radiation damage

    NASA Technical Reports Server (NTRS)

    Goradia, C.; Vaughn, J.; Baraona, C. R.

    1980-01-01

    Results of computer calculations using a one dimensional model of the silicon tandem junction solar cell with both front and back current collection are presented. Using realistically achievable geometrical and material parameters, the model predicts that with base widths of 50 micrometers and 100 micrometers and base resistivities between 1 ohm/cm and 20 ohm/cm, beginning of life efficiencies of 14% to 17% and end of life efficiencies of 12% to 14%, after about seven years in synchronous orbit, can be obtained.

  17. Fracture toughness of solid oxide fuel cell anode substrates determined by a double-torsion technique

    NASA Astrophysics Data System (ADS)

    Pećanac, G.; Wei, J.; Malzbender, J.

    2016-09-01

    Planar solid oxide fuel cell anode substrates are exposed to high mechanical loads during assembly, start-up, steady-state operation and thermal cycling. Hence, characterization of mechanical stability of anode substrates under different oxidation states and at relevant temperatures is essential to warrant a reliable operation of solid oxide fuel cells. As a basis for mechanical assessment of brittle supports, two most common anode substrate material variants, NiO-3YSZ and NiO-8YSZ, were analyzed in this study with respect to their fracture toughness at room temperature and at a typical stack operation temperature of 800 °C. The study considered both, oxidized and reduced materials' states, where also an outlook is given on the behavior of the re-oxidized state that might be induced by malfunctions of sealants or other functional components. Aiming at the improvement of material's production, different types of warm pressed and tape cast NiO-8YSZ substrates were characterized in oxidized and reduced states. Overall, the results confirmed superior fracture toughness of 3YSZ compared to 8YSZ based composites in the oxidized state, whereas in the reduced state 3YSZ based composites showed similar fracture toughness at room temperature, but a higher value at 800 °C compared to 8YSZ based composites. Complementary microstructural analysis aided the interpretation of mechanical characterization.

  18. Rapid double-dye-layer coating for dye-sensitized solar cells using a new method.

    PubMed

    Jung, Cho-long; Han, Chi-Hwan; Moon, Doo Kyung; Jun, Yongseok

    2014-10-01

    Intensive research with the specific aim of developing inexpensive renewable energy sources is currently being undertaken. In dye-sensitized solar cell (DSSC) production, the most time-consuming process is coating the dye on working electrodes: absorption of ruthenium-based dyes [e.g., N719=bis(trtrabutylammonium)-cis-di(thiocyanato)-N,N'-bis(4-carboxylato-4'-carboxylic acid-2,2'-bipyridine) ruthenium(II)] on a photoanode takes a long time. We report a simple dye-coating method using a mixed solvent of ethylene glycol (EG) and glycerol (Gly). According to our experiments, dye-coating time can be reduced to 5 min from several hours. Maximum performance was obtained with an EG/Gly ratio of 1:1. This mixture of solvents gave a performance of 9.1%. Furthermore, the viscous solvent system could control coating depth; positioning dye coatings to a specific depth was rapid and facile. A cell containing two different dyes (N719+black dye) had an efficiency of 9.4%. PMID:25154611

  19. Purification of recombinant proteins from mammalian cell culture using a generic double-affinity chromatography scheme.

    PubMed

    Cass, Brian; Pham, Phuong Lan; Kamen, Amine; Durocher, Yves

    2005-03-01

    Transient transfection of mammalian cells has proven to be a useful technique for the rapid production of recombinant proteins because of its ability to produce milligram quantities within 2 weeks following cloning of their corresponding cDNA. This rapid production also requires a fast and efficient purification scheme that can be applied generically, typically through the use of affinity tags such as the polyhistidine-tag for capture by immobilized metal-affinity chromatography (IMAC) or the Strep-tag II, which binds to the StrepTactin affinity ligand. However, one-step purification using either of these tags has disadvantages in terms of yield, elution conditions, and purity. Here, we show that the addition of both Strep-tag-II and (His)(8) to the C-terminal of r-proteins allows efficient purification by consecutive IMAC and StrepTactin affinity. This approach has been successfully demonstrated using the intracellular protein DsRed, as well as two secreted proteins, secreted alkaline phosphatase (SEAP) and vascular endothelial growth factor (VEGF), all produced by transient transfection of HEK293-EBNA1 cells in medium supplemented with bovine calf serum. All proteins were purified to >99% homogeneity with yields varying from 29 to 81%. PMID:15721774

  20. Detection of space radiation-induced double strand breaks as a track in cell nucleus

    SciTech Connect

    Ohnishi, Takeo; Takahashi, Akihisa; Nagamatsu, Aiko; Omori, Katsunori; Suzuki, Hiromi; Shimazu, Toru; Ishioka, Noriaki

    2009-12-18

    To identify DNA damage induced by space radiations such as the high linear energy transfer (LET) particles, phospho-H2AX ({gamma}H2AX) foci formation was analyzed in human cells frozen in an International Space Station freezer for 133 days. After recovering the frozen sample to the earth, the cells were cultured for 30 min, and then fixed. Here we show a track of {gamma}H2AX positive foci in them by immuno-cytochemical methods. It is suggested that space radiations, especially high LET particles, induced DSBs as a track. From the formation of the tracks in nuclei, exposure dose rate was calculated to be 0.7 mSv per day as relatively high-energy space radiations of Fe-ions (500 MeV/u, 200 keV/{mu}m). From the physical dosimetry with CR-39 plastic nuclear track detectors and thermo-luminescent dosimeters, dose rate was 0.5 mSv per day. These values the exposed dose rate were similar between biological and physical dosimetries.

  1. DYRK1A: the double-edged kinase as a protagonist in cell growth and tumorigenesis

    PubMed Central

    Fernández-Martínez, P; Zahonero, C; Sánchez-Gómez, P

    2015-01-01

    DYRK1A (dual-specificity tyrosine-regulated kinase 1A) is a kinase with multiple implications for embryonic development, especially in the nervous system where it regulates the balance between proliferation and differentiation of neural progenitors. The DYRK1A gene is located in the Down syndrome critical region and may play a significant role in the developmental brain defects, early neurodegeneration, and cancer susceptibility of individuals with this syndrome. DYRK1A is also expressed in adults, where it might participate in the regulation of cell cycle, survival, and tumorigenesis, thus representing a potential therapeutic target for certain types of cancer. However, the final readout of DYRK1A overexpression or inhibition depends strongly on the cellular context, as it has both tumor suppressor and oncogenic activities. Here, we will discuss the functions and substrates of DYRK1A associated with the control of cell growth and tumorigenesis with a focus on the potential use of DYRK1A inhibitors in cancer therapy. PMID:27308401

  2. Hematopoietic Stem Cells from Ts65Dn Mice Are Deficient in the Repair of DNA Double-Strand Breaks.

    PubMed

    Wang, Yingying; Chang, Jianhui; Shao, Lijian; Feng, Wei; Luo, Yi; Chow, Marie; Du, Wei; Meng, Aimin; Zhou, Daohong

    2016-06-01

    Down syndrome (DS) is a genetic disorder caused by the presence of an extra partial or whole copy of chromosome 21. In addition to musculoskeletal and neurodevelopmental abnormalities, children with DS exhibit various hematologic disorders and have an increased risk of developing acute lymphoblastic leukemia and acute megakaryocytic leukemia. Using the Ts65Dn mouse model, we investigated bone marrow defects caused by trisomy for 132 orthologs of the genes on human chromosome 21. The results showed that, although the total bone marrow cellularity as well as the frequency of hematopoietic progenitor cells (HPCs) was comparable between Ts65Dn mice and their age-matched euploid wild-type (WT) control littermates, human chromosome 21 trisomy led to a significant reduction in hematopoietic stem cell (HSC) numbers and clonogenic function in Ts65Dn mice. We also found that spontaneous DNA double-strand breaks (DSBs) were significantly increased in HSCs from the Ts65Dn mice, which was correlated with the significant reduction in HSC clonogenic activity compared to those from WT controls. Moreover, analysis of the repair kinetics of radiation-induced DSBs revealed that HSCs from Ts65Dn mice were less proficient in DSB repair than the cells from WT controls. This deficiency was associated with a higher sensitivity of Ts65Dn HSCs to radiation-induced suppression of HSC clonogenic activity than that of euploid HSCs. These findings suggest that an additional copy of genes on human chromosome 21 may selectively impair the ability of HSCs to repair DSBs, which may contribute to DS-associated hematological abnormalities and malignancies. PMID:27243896

  3. Programmed cell death 1 and Helios distinguish TCR-αβ+ double negative (CD4-CD8-) T cells that derive from self-reactive CD8 T cells1

    PubMed Central

    Rodríguez-Rodríguez, Noé; Apostolidis, Sokratis A.; Penaloza-MacMaster, Pablo; Manuel Martín Villa, José; Barouch, Dan H.; Tsokos, George C.; Crispín, José C.

    2015-01-01

    TCR-αβ+ double negative (DN; CD4-CD8-) T cells represent a poorly understood cellular subset suggested to contribute to the pathogenesis of the autoimmune disease systemic lupus erythematosus. DN T cells have been proposed to derive from CD8+ cells. However, the conditions that govern the loss of CD8 expression after antigen encounter are unknown. Here we tracked the fate of CD8 T cells from transgenic TCR mice exposed to their cognate antigens as self or in the context of infection. We demonstrate that CD8 T cells lose CD8 expression and become DN only when cognate antigen is sensed as self. This process is restricted to tissues where the antigen is present. We also show that DN T cells derived from self-reactive CD8 cells express the inhibitory molecules PD-1 and Helios. These molecules identify a subset of DN T cells in normal mice. A similar population expands when CD8 T cells from repertoires enriched in self-reactive cells (Aire-deficient) are transferred into cognate hosts. Collectively, our data suggest that a subset of DN T cells, identified by the expression of PD-1 and Helios, represent self-reactive cells. Our results provide an explanation for the origin of DN T cells and introduce CD8 loss as a process associated to self-antigen encounter. PMID:25825451

  4. Optimization of amorphous silicon double junction solar cells for an efficient photoelectrochemical water splitting device based on a bismuth vanadate photoanode.

    PubMed

    Han, Lihao; Abdi, Fatwa F; Perez Rodriguez, Paula; Dam, Bernard; van de Krol, Roel; Zeman, Miro; Smets, Arno H M

    2014-03-01

    A photoelectrochemical water splitting device (PEC-WSD) was designed and fabricated based on cobalt-phosphate-catalysed and tungsten-gradient-doped bismuth vanadate (W:BiVO4) as the photoanode. A simple and cheap hydrogenated amorphous silicon (a-Si:H) double junction solar cell has been used to provide additional bias. The advantage of using thin film silicon (TF-Si) based solar cells is that this photovoltaic (PV) technology meets the crucial requirements for the PV component in PEC-WSDs based on W:BiVO4 photoanodes. TF-Si PV devices are stable in aqueous solutions, are manufactured by simple and cheap fabrication processes and their spectral response, voltage and current density show an excellent match with the photoanode. This paper is mainly focused on the optimization of the TF-Si solar cell with respect to the remaining solar spectrum transmitted through the W:BiVO4 photoanode. The current matching between the top and bottom cells is studied and optimized by varying the thickness of the a-Si:H top cell. We support the experimental optimization of the current balance between the two sub-cells with simulations of the PV devices. In addition, the impact of the light induced degradation of the a-Si:H double junction, the so-called Staebler-Wronski Effect (SWE), on the performance of the PEC-WSD has been studied. The light soaking experiments on the a-Si:H/a-Si:H double junctions over 1000 hours show that the efficiency of a stand-alone a-Si:H/a-Si:H double junction cell is significantly reduced due to the SWE. Nevertheless, the SWE has a significantly smaller effect on the performance of the PEC-WSD. PMID:24452785

  5. Slip Correction Measurements of Certified PSL Nanoparticles Using a Nanometer Differential Mobility Analyzer (Nano-DMA) for Knudsen Number From 0.5 to 83

    PubMed Central

    Kim, Jung Hyeun; Mulholland, George W.; Kukuck, Scott R.; Pui, David Y. H.

    2005-01-01

    The slip correction factor has been investigated at reduced pressures and high Knudsen number using polystyrene latex (PSL) particles. Nano-differential mobility analyzers (NDMA) were used in determining the slip correction factor by measuring the electrical mobility of 100.7 nm, 269 nm, and 19.90 nm particles as a function of pressure. The aerosol was generated via electrospray to avoid multiplets for the 19.90 nm particles and to reduce the contaminant residue on the particle surface. System pressure was varied down to 8.27 kPa, enabling slip correction measurements for Knudsen numbers as large as 83. A condensation particle counter was modified for low pressure application. The slip correction factor obtained for the three particle sizes is fitted well by the equation: C = 1 + Kn (α + β exp(−γ/Kn)), with α = 1.165, β = 0.483, and γ = 0.997. The first quantitative uncertainty analysis for slip correction measurements was carried out. The expanded relative uncertainty (95 % confidence interval) in measuring slip correction factor was about 2 % for the 100.7 nm SRM particles, about 3 % for the 19.90 nm PSL particles, and about 2.5 % for the 269 nm SRM particles. The major sources of uncertainty are the diameter of particles, the geometric constant associated with NDMA, and the voltage. PMID:27308102

  6. Calculation of friction coefficient and analysis of fluid flow in a stepped micro-channel for wide range of Knudsen number using Lattice Boltzmann (MRT) method

    NASA Astrophysics Data System (ADS)

    Bakhshan, Younes; Omidvar, Alireza

    2015-12-01

    Micro scale gas flows have attracted significant research interest in the last two decades. In this research, the fluid flow of gases in a stepped micro-channel has been conducted. Wide range of Knudsen number has been implemented using the Lattice Boltzmann (MRT) method in this study. A modified second-order slip boundary condition and a Bosanquet-type effective viscosity are used to consider the velocity slip at the boundaries and to cover the slip and transition regimes of flow to obtain an accurate simulation of rarefied gases. The flow specifications such as pressure loss, velocity profile, stream lines and friction coefficient at different conditions have been presented. The results show, good agreement with available experimental data. The calculation shows, that the friction coefficient decreases with increasing the Knudsen number and stepping the micro-channel has an inverse effect on the friction coefficient value. Furthermore, a new correlation is suggested for calculation of the friction coefficient in the stepped micro-channel flows as below;

  7. The origin of a large apparent tortuosity factor for the Knudsen diffusion inside monoliths of a samaria-alumina aerogel catalyst: a diffusion NMR study.

    PubMed

    Mueller, R; Zhang, S; Klink, M; Bäumer, M; Vasenkov, S

    2015-11-01

    Pulsed field gradient (PFG) NMR was applied to measure tortuosity factors for carbon dioxide diffusion in the Knudsen and gas regimes inside monoliths of a samaria-alumina aerogel catalyst, a high porosity material containing micropores in addition to meso- and macropores. The apparent tortuosity factor obtained from PFG NMR measurements for the Knudsen diffusion in the meso- and macropores of the catalyst has an unexpectedly large value of approximately 6 if carbon dioxide adsorption in the micropores and other types of surface adsorption sites of the catalyst is ignored. At the same time, the corresponding apparent tortuosity factor in the gas regime was found to be around 2. Application of a proposed model which describes fast molecular exchange between the surface adsorption sites and the main pore volume of the catalyst yields corrected tortuosity factors which depend only on the pore system geometry. Using this model, the corrected tortuosity factors were found to be around 2 for both diffusion regimes, in agreement with the expectations based on a high porosity of the studied catalyst. PMID:26426141

  8. Compilation and evaluation of gas-phase diffusion coefficients of reactive trace gases in the atmosphere: volume 2. Organic compounds and Knudsen numbers for gas uptake calculations

    NASA Astrophysics Data System (ADS)

    Tang, M. J.; Shiraiwa, M.; Pöschl, U.; Cox, R. A.; Kalberer, M.

    2015-02-01

    Diffusion of organic vapours to the surface of aerosol or cloud particles is an important step for the formation and transformation of atmospheric particles. So far, however, a database of gas phase diffusion coefficients for organic compounds of atmospheric interest has not been available. In this work we have compiled and evaluated gas phase diffusivities (pressure-independent diffusion coefficients) of organic compounds reported by previous experimental studies, and we compare the measurement data to estimates obtained with Fuller's semi-empirical method. The difference between measured and estimated diffusivities are mostly < 10%. With regard to gas-particle interactions, different gas molecules, including both organic and inorganic compounds, exhibit similar Knudsen numbers (Kn) although their gas phase diffusivities may vary over a wide range. Knudsen numbers of gases with unknown diffusivity can be approximated by a simple function of particle diameter and pressure and can be used to characterize the influence of diffusion on gas uptake by aerosol or cloud particles. We use a kinetic multi-layer model of gas-particle interaction to illustrate the effects of gas phase diffusion on the condensation of organic compounds with different volatilities. The results show that gas-phase diffusion can play a major role in determining the growth of secondary organic aerosol particles by condensation of low-volatility organic vapours.

  9. Surface stress, initial stress and Knudsen-dependent flow velocity effects on the electro-thermo nonlocal wave propagation of SWBNNTs

    NASA Astrophysics Data System (ADS)

    Ghorbanpour Arani, A.; Roudbari, M. A.

    2014-11-01

    This paper investigates the electro-thermal nonlocal wave propagation of fluid-conveying single-walled Boron Nitride nanotubes (SWBNNTs) using nonlocal piezoelasticity with surface stress, initial stress and Knudsen-dependent flow velocity effect. SWBNNT is embedded in a vicsoelastic medium which is simulated as visco-Pasternak foundation. Using Euler-Bernoulli beam (EBB) model, Hamilton's principle and nonlocal piezoelasticity theory, the higher order governing equation is derived. A detailed parametric study is conducted, focusing on the combined effects of the electric parameters, viscoelastic medium, initial stress, surface stress, Knudsen number (Kn) and small scale on the wave propagation behaviour of the fluid-conveying SWBNNT. The results show that for smaller values of wave number the dispersion relation for different fluid viscosities seems to be similar. At the higher values of wave numbers, increase in the wave frequency values is remarkable due to increase in fluid viscosity. The electric field as a smart controller, surface effect, initial stress, temperature change and slip velocity effect have significant role on the wave frequency. The results of this work is hoped to be of use in design and manufacturing of smart MEMS/NEMS in advanced medical applications such as drug delivery systems with great applications in biomechanics.

  10. Targeted gene exchange in plant cells mediated by a zinc finger nuclease double cut.

    PubMed

    Schneider, Katja; Schiermeyer, Andreas; Dolls, Anja; Koch, Natalie; Herwartz, Denise; Kirchhoff, Janina; Fischer, Rainer; Russell, Sean M; Cao, Zehui; Corbin, David R; Sastry-Dent, Lakshmi; Ainley, W Michael; Webb, Steven R; Schinkel, Helga; Schillberg, Stefan

    2016-04-01

    Genome modification by homology-directed repair (HDR) is an attractive tool for the controlled genetic manipulation of plants. Here, we report the HDR-mediated gene exchange of expression cassettes in tobacco BY-2 cells using a designed zinc finger nuclease (ZFN). The target contained a 7-kb fragment flanked by two ZFN cutting sites. That fragment was replaced with a 4-kb donor cassette, which integrates gene markers for selection (kanamycin resistance) and for scoring targeting (red fluorescent protein, RFP). Candidates resulting from cassette exchange were identified by molecular analysis of calli generated by transformation via direct DNA delivery. The precision of HDR-mediated donor integration was evaluated by Southern blot analysis, sequencing of the integration locus and analysis of RFP fluorescence by flow cytometry. Screening of 1326 kanamycin-resistant calli yielded 18 HDR events, 16 of which had a perfect cassette exchange at the insert junction and 13 of which produced functional RFP. Our results demonstrate that ZFN-based HDR can be used for high frequency, precise, targeted exchange of fragments of sizes that are commercially relevant in plants. PMID:26426390

  11. DNA Double-Strand Breaks Induced by Cavitational Mechanical Effects of Ultrasound in Cancer Cell Lines

    PubMed Central

    Furusawa, Yukihiro; Fujiwara, Yoshisada; Campbell, Paul; Zhao, Qing-Li; Ogawa, Ryohei; Ali Hassan, Mariame; Tabuchi, Yoshiaki; Takasaki, Ichiro; Takahashi, Akihisa; Kondo, Takashi

    2012-01-01

    Ultrasonic technologies pervade the medical field: as a long established imaging modality in clinical diagnostics; and, with the emergence of targeted high intensity focused ultrasound, as a means of thermally ablating tumours. In parallel, the potential of [non-thermal] intermediate intensity ultrasound as a minimally invasive therapy is also being rigorously assessed. Here, induction of apoptosis in cancer cells has been observed, although definitive identification of the underlying mechanism has thus far remained elusive. A likely candidate process has been suggested to involve sonochemical activity, where reactive oxygen species (ROS) mediate the generation of DNA single-strand breaks. Here however, we provide compelling new evidence that strongly supports a purely mechanical mechanism. Moreover, by a combination of specific assays (neutral comet tail and staining for γH2AX foci formation) we demonstrate for the first time that US exposure at even moderate intensities exhibits genotoxic potential, through its facility to generate DNA damage across multiple cancer lines. Notably, colocalization assays highlight that ionizing radiation and ultrasound have distinctly different signatures to their respective γH2AX foci formation patterns, likely reflecting the different stress distributions that initiated damage formation. Furthermore, parallel immuno-blotting suggests that DNA-PKcs have a preferential role in the repair of ultrasound-induced damage. PMID:22235259

  12. Induction of DNA Double-Strand Breaks and Cellular Migration Through Bystander Effects in Cells Irradiated With the Slit-Type Microplanar Beam of the Spring-8 Synchrotron

    SciTech Connect

    Kashino, Genro Kondoh, Takeshi; Nariyama, Nobuteru; Umetani, Keiji; Ohigashi, Takuji; Shinohara, Kunio; Kurihara, Ai; Fukumoto, Manabu; Tanaka, Hiroki; Maruhashi, Akira; Suzuki, Minoru; Kinashi, Yuko; Liu Yong; Masunaga, Shin-ichiro; Watanabe, Masami; Ono, Koji

    2009-05-01

    Purpose: To determine whether glioma cells irradiated with a microplanar X-ray beam exert bystander effects. Methods and Materials: Microplanar beam irradiation of glioma cells in vitro was done using the SPring-8 synchrotron radiation facility. The amount of DNA double-strand breaks (dsbs) was measured by the fluorescence intensity of phosphorylated H2AX or the number of 53BP1 foci. The dose distribution in a cell population exposed to a single microplanar beam was determined by the amount of phosphorylated H2AX-positive cells. Bystander effects were determined by counting the number of 53BP1 foci in nonirradiated cells treated with conditioned medium from cultures of irradiated cells. Results: More DNA dsbs were detected in cells adjacent to an area irradiated by the single beam than in cells in distant, nonirradiated areas as a result of bystander effects caused by scattered X-rays and DNA dsbs. In support of this, more 53BP1 foci were observed in nonirradiated, conditioned medium-treated cells than in control cells (i.e., cells not treated with irradiation or conditioned medium). These results suggest that DNA dsbs were induced in nonirradiated cells by soluble factors in the culture medium. In addition, we observed cellular migration into areas irradiated with peak doses, suggesting that irradiated cells send signals that cause nonirradiated cells to migrate toward damaged cells. Conclusions: Bystander effects are produced by factors secreted as a result of slit-type microplanar X-ray beam irradiation.

  13. B-cell lymphomas with concurrent MYC and BCL2 abnormalities other than translocations behave similarly to MYC/BCL2 double-hit lymphomas.

    PubMed

    Li, Shaoying; Seegmiller, Adam C; Lin, Pei; Wang, Xuan J; Miranda, Roberto N; Bhagavathi, Sharathkumar; Medeiros, L Jeffrey

    2015-02-01

    Large B-cell lymphomas with IGH@BCL2 and MYC rearrangement, known as double-hit lymphoma (DHL), are clinically aggressive neoplasms with a poor prognosis. Some large B-cell lymphomas have concurrent abnormalities of MYC and BCL2 other than coexistent translocations. Little is known about patients with these lymphomas designated here as atypical DHL. We studied 40 patients of atypical DHL including 21 men and 19 women, with a median age of 60 years. Nine (23%) patients had a history of B-cell non-Hodgkin lymphoma. There were 30 diffuse large B-cell lymphoma (DLBCL), 7 B-cell lymphoma, unclassifiable, with features intermediate between DLBCL and Burkitt lymphoma, and 3 DLBCL with coexistent follicular lymphoma. CD10, BCL2, and MYC were expressed in 28/39 (72%), 33/35 (94%), and 14/20 (70%) cases, respectively. Patients were treated with standard (n=14) or more aggressive chemotherapy regimens (n=17). We compared the atypical DHL group with 76 patients with DHLand 35 patients with DLBCL lacking MYC and BCL2 abnormalities. The clinicopathologic features and therapies were similar between patients with atypical and typical DHL. The overall survival of patients with atypical double-hit lymphoma was similar to that of patients with double-hit lymphoma (P=0.47) and significantly worse than that of patients with DLBCL with normal MYC and BCL2 (P=0.02). There were some minor differences. Cases of atypical double-hit lymphoma more often have DLBCL morphology (P<0.01), less frequently expressed CD10 (P<0.01), and patients less often had an elevated serum lactate dehydrogenase level (P=0.01). In aggregate, these results support expanding the category of MYC/BCL2 DHL to include large B-cell lymphomas with coexistent MYC and BCL2 abnormalities other than concurrent translocations. PMID:25103070

  14. Double transmission peaks electromagnetically induced transparency induced by simultaneously exciting the electric and magnetic resonance in one unit cell

    NASA Astrophysics Data System (ADS)

    Liu, Si-Yuan; Zheng, Bu-Sheng; Li, Hai-Ming; Liu, Xiao-Chun; Liu, Shao-Bin

    2015-08-01

    In this paper, we investigate a metamaterial formed by a planar array of a metallic L-shaped structure and a cut wire (CW), which behaves as an analogue of the electromagnetically induced transparency (EIT). The double transmission peaks are formed by the destructive interference of two bright-modes and a quasi-dark mode. The two bright-modes are respectively excited by the L-shaped structure and CW. The unit structure itself performs a quasi-dark mode. The group refractive indexes are over 20 in the first transmission peak, and 117 in the second transmission peak, thus offering potential applications in slow light devices. Finally, all the above characteristics are achieved in just one simple unit cell. Project supported by the Chinese Specialized Research Fund for the Doctoral Program of Higher Education, China (Grant No. 20123218110017), the National Natural Science Foundation of China (Grant Nos. 61307052 and 61471368), the Foundation of Aeronautical Science, China (Grant No. 20121852030), and the Fundamental Research Funds for the Central Universities (Grant No. kfjj20150407).

  15. Induction and repair of zinc-finger nuclease-targeted double-strand breaks in Caenorhabditis elegans somatic cells

    PubMed Central

    Morton, Jason; Davis, M. Wayne; Jorgensen, Erik M.; Carroll, Dana

    2006-01-01

    Zinc-finger nucleases are chimeric proteins consisting of engineered zinc-finger DNA-binding motifs attached to an endonuclease domain. These proteins can induce site-specific DNA double-strand breaks in genomic DNA, which are then substrates for cellular repair mechanisms. Here, we demonstrate that engineered zinc-finger nucleases function effectively in somatic cells of the nematode Caenorhabditis elegans. Although gene-conversion events were indistinguishable from uncut DNA in our assay, nonhomologous end joining resulted in mutations at the target site. A synthetic target on an extrachromosomal array was targeted with a previously characterized nuclease, and an endogenous genomic sequence was targeted with a pair of specifically designed nucleases. In both cases, ≈20% of the target sites were mutated after induction of the corresponding nucleases. Alterations in the extrachromosomal targets were largely products of end-filling and blunt ligation. By contrast, alterations in the chromosomal target were mostly deletions. We interpret these differences to reflect the abundance of homologous templates present in the extrachromosomal arrays versus the paucity of such templates for repair of chromosomal breaks. In addition, we find evidence for the involvement of error-prone DNA synthesis in both homologous and nonhomologous pathways of repair. DNA ligase IV is required for efficient end joining, particularly of blunt ends. In its absence, a secondary end-joining pathway relies more heavily on microhomologies in producing deletions. PMID:17060623

  16. A double-blind, randomized, multicenter phase 2 study of prasugrel versus placebo in adult patients with sickle cell disease

    PubMed Central

    2013-01-01

    Background Platelet activation has been implicated in the pathogenesis of sickle cell disease (SCD) suggesting antiplatelet agents may be therapeutic. To evaluate the safety of prasugrel, a thienopyridine antiplatelet agent, in adult patients with SCD, we conducted a double-blind, randomized, placebo-controlled study. Methods The primary endpoint, safety, was measured by hemorrhagic events requiring medical intervention. Patients were randomized to prasugrel 5 mg daily (n = 41) or placebo (n = 21) for 30 days. Platelet function by VerifyNow® P2Y12 and vasodilator-stimulated phosphoprotein assays at days 10 and 30 were significantly inhibited in prasugrel- compared with placebo-treated SCD patients. Results There were no hemorrhagic events requiring medical intervention in either study arm. Mean pain rate (percentage of days with pain) and intensity in the prasugrel arm were decreased compared with placebo. However, these decreases did not reach statistical significance. Platelet surface P-selectin and plasma soluble P-selectin, biomarkers of in vivo platelet activation, were significantly reduced in SCD patients receiving prasugrel compared with placebo. In sum, prasugrel was well tolerated and not associated with serious hemorrhagic events. Conclusions Despite the small size and short duration of this study, there was a decrease in platelet activation biomarkers and a trend toward decreased pain. PMID:23414938

  17. Molecular Process Producing Oncogene Fusion in Lung Cancer Cells by Illegitimate Repair of DNA Double-Strand Breaks

    PubMed Central

    Seki, Yoshitaka; Mizukami, Tatsuji; Kohno, Takashi

    2015-01-01

    Constitutive activation of oncogenes by fusion to partner genes, caused by chromosome translocation and inversion, is a critical genetic event driving lung carcinogenesis. Fusions of the tyrosine kinase genes ALK (anaplastic lymphoma kinase), ROS1 (c-ros oncogene 1), or RET (rearranged during transfection) occur in 1%–5% of lung adenocarcinomas (LADCs) and their products constitute therapeutic targets for kinase inhibitory drugs. Interestingly, ALK, RET, and ROS1 fusions occur preferentially in LADCs of never- and light-smokers, suggesting that the molecular mechanisms that cause these rearrangements are smoking-independent. In this study, using previously reported next generation LADC genome sequencing data of the breakpoint junction structures of chromosome rearrangements that cause oncogenic fusions in human cancer cells, we employed the structures of breakpoint junctions of ALK, RET, and ROS1 fusions in 41 LADC cases as “traces” to deduce the molecular processes of chromosome rearrangements caused by DNA double-strand breaks (DSBs) and illegitimate joining. We found that gene fusion was produced by illegitimate repair of DSBs at unspecified sites in genomic regions of a few kb through DNA synthesis-dependent or -independent end-joining pathways, according to DSB type. This information will assist in the understanding of how oncogene fusions are generated and which etiological factors trigger them. PMID:26437441

  18. Protease-Resistant and Cell-Permeable Double-Stapled Peptides Targeting the Rab8a GTPase.

    PubMed

    Cromm, Philipp M; Spiegel, Jochen; Küchler, Philipp; Dietrich, Laura; Kriegesmann, Julia; Wendt, Mathias; Goody, Roger S; Waldmann, Herbert; Grossmann, Tom N

    2016-08-19

    Small GTPases comprise a family of highly relevant targets in chemical biology and medicinal chemistry research and have been considered "undruggable" due to the persisting lack of effective synthetic modulators and suitable binding pockets. As molecular switches, small GTPases control a multitude of pivotal cellular functions, and their dysregulation is associated with many human diseases such as various forms of cancer. Rab-GTPases represent the largest subfamily of small GTPases and are master regulators of vesicular transport interacting with various proteins via flat and extensive protein-protein interactions (PPIs). The only reported synthetic inhibitor of a PPI involving an activated Rab GTPase is the hydrocarbon stapled peptide StRIP3. However, this macrocyclic peptide shows low proteolytic stability and cell permeability. Here, we report the design of a bioavailable StRIP3 analogue that harbors two hydrophobic cross-links and exhibits increased binding affinity, combined with robust cellular uptake and extremely high proteolytic stability. Localization experiments reveal that this double-stapled peptide and its target protein Rab8a accumulate in the same cellular compartments. The reported approach offers a strategy for the implementation of biostability into conformationally constrained peptides while supporting cellular uptake and target affinity, thereby conveying drug-like properties. PMID:27336832

  19. The H19/let-7 double-negative feedback loop contributes to glucose metabolism in muscle cells

    PubMed Central

    Gao, Yuan; Wu, Fuju; Zhou, Jichun; Yan, Lei; Jurczak, Michael J.; Lee, Hui-Young; Yang, Lihua; Mueller, Martin; Zhou, Xiao-Bo; Dandolo, Luisa; Szendroedi, Julia; Roden, Michael; Flannery, Clare; Taylor, Hugh; Carmichael, Gordon G.; Shulman, Gerald I.; Huang, Yingqun

    2014-01-01

    The H19 lncRNA has been implicated in development and growth control and is associated with human genetic disorders and cancer. Acting as a molecular sponge, H19 inhibits microRNA (miRNA) let-7. Here we report that H19 is significantly decreased in muscle of human subjects with type-2 diabetes and insulin resistant rodents. This decrease leads to increased bioavailability of let-7, causing diminished expression of let-7 targets, which is recapitulated in vitro where H19 depletion results in impaired insulin signaling and decreased glucose uptake. Furthermore, acute hyperinsulinemia downregulates H19, a phenomenon that occurs through PI3K/AKT-dependent phosphorylation of the miRNA processing factor KSRP, which promotes biogenesis of let-7 and its mediated H19 destabilization. Our results reveal a previously undescribed double-negative feedback loop between sponge lncRNA and target miRNA that contributes to glucose regulation in muscle cells. PMID:25399420

  20. An ultrasensitive squamous cell carcinoma antigen biosensing platform utilizing double-antibody single-channel amplification strategy.

    PubMed

    Ren, Xiang; Wu, Dan; Wang, Yuhuan; Zhang, Yunhui; Fan, Dawei; Pang, Xuehui; Li, Yueyun; Du, Bin; Wei, Qin

    2015-10-15

    A novel electrochemical immunosensor was developed for ultrasensitive detection of squamous cell carcinoma antigen (SCCA), which was based on the double-antibody single-channel amplification strategy. For the first time, human immunoglobulin antibody (anti-HIgG) was used as the supporting framework to amplify the loading quantity of SCCA antibody (anti-SCCA). In this strategy, SCCA can be detected without using mesoporous nanometers to amplify the signal. In addition, Pd icosahedrons were first used as the connecter to immobilize the antibodies and strengthen the sensitivity. Only one touch point exists under the limited condition between a sphere and another shape in geometry, thus the Pd icosahedron is an excellent candidate as the role of connecter. Gold nanoparticles (Au NPs) decorated with mercapto-functionalized graphene sheets (Au@GS) were synthesized as the transducing materials. The fabricated immunosensor exhibited an excellent detection limit of 2.8 pg/mL and wide linear range of 0.01-5 ng/mL. This kind of immunosensor would provide a potential application in clinical diagnosis. PMID:25982722

  1. Isolation of Human Monocytes by Double Gradient Centrifugation and Their Differentiation to Macrophages in Teflon-coated Cell Culture Bags

    PubMed Central

    Menck, Kerstin; Behme, Daniel; Pantke, Mathias; Reiling, Norbert; Binder, Claudia; Pukrop, Tobias; Klemm, Florian

    2014-01-01

    Human macrophages are involved in a plethora of pathologic processes ranging from infectious diseases to cancer. Thus they pose a valuable tool to understand the underlying mechanisms of these diseases. We therefore present a straightforward protocol for the isolation of human monocytes from buffy coats, followed by a differentiation procedure which results in high macrophage yields. The technique relies mostly on commonly available lab equipment and thus provides a cost and time effective way to obtain large quantities of human macrophages. Briefly, buffy coats from healthy blood donors are subjected to a double density gradient centrifugation to harvest monocytes from the peripheral blood. These monocytes are then cultured in fluorinated ethylene propylene (FEP) Teflon-coated cell culture bags in the presence of macrophage colony-stimulating factor (M-CSF). The differentiated macrophages can be easily harvested and used for subsequent studies and functional assays. Important methods for quality control and validation of the isolation and differentiation steps will be highlighted within the protocol. In summary, the protocol described here enables scientists to routinely and reproducibly isolate human macrophages without the need for cost intensive tools. Furthermore, disease models can be studied in a syngeneic human system circumventing the use of murine macrophages. PMID:25226391

  2. Fully 3D Particle-in-Cell Simulation of Double Post-Hole Convolute on PTS Facility

    NASA Astrophysics Data System (ADS)

    Zhao, Hailong; Dong, Ye; Zhou, Haijing; Zou, Wenkang; Institute of Fluid Physics Collaboration; Institute of Applied Physics; Computational Mathematics Collaboration

    2015-11-01

    In order to get better understand of energy transforming and converging process during High Energy Density Physics (HEDP) experiments, fully 3D particle-in-cell (PIC) simulation code NEPTUNE3D was used to provide numerical approach towards parameters which could hardly be acquired through diagnostics. Cubic region (34cm × 34cm × 18cm) including the double post-hole convolute (DPHC) on the primary test stand (PTS) facility was chosen to perform a series of fully 3D PIC simulations, calculating ability of codes were tested and preliminary simulation results about DPHC on PTS facility were discussed. Taking advantages of 3D simulation codes and large-scale parallel computation, massive data (~ 250GB) could be acquired in less than 5 hours and clear process of current transforming and electron emission in DPHC were demonstrated with the help of visualization tools. Cold-chamber tests were performed during which only cathode electron emission was considered without temperature rise or ion emission, current loss efficiency was estimated to be 0.46% ~ 0.48% by comparisons between output magnetic field profiles with or without electron emission. Project supported by the National Natural Science Foundation of China (Grant No. 11205145, 11305015, 11475155).

  3. Changes in chromatin structure and mobility in living cells at sites of DNA double-strand breaks.

    PubMed

    Kruhlak, Michael J; Celeste, Arkady; Dellaire, Graham; Fernandez-Capetillo, Oscar; Müller, Waltraud G; McNally, James G; Bazett-Jones, David P; Nussenzweig, André

    2006-03-13

    The repair of DNA double-strand breaks (DSBs) is facilitated by the phosphorylation of H2AX, which organizes DNA damage signaling and chromatin remodeling complexes in the vicinity of the lesion. The disruption of DNA integrity induces an alteration of chromatin architecture that has been proposed to activate the DNA damage transducing kinase ataxia telangiectasia mutated. However, little is known about the physical properties of damaged chromatin. In this study, we use a photoactivatable version of GFP-tagged histone H2B to examine the mobility and structure of chromatin containing DSBs in living cells. We find that chromatin containing DSBs exhibits limited mobility but undergoes an energy-dependent local expansion immediately after DNA damage. The localized expansion observed in real time corresponds to a 30-40% reduction in the density of chromatin fibers in the vicinity of DSBs, as measured by energy-filtering transmission electron microscopy. The observed opening of chromatin occurs independently of H2AX and ATM. We propose that localized adenosine triphosphate-dependent decondensation of chromatin at DSBs establishes an accessible subnuclear environment that facilitates DNA damage signaling and repair. PMID:16520385

  4. Double-chamber microbial fuel cell with a non-platinum-group metal Fe-N-C cathode catalyst.

    PubMed

    Santoro, Carlo; Serov, Alexey; Narvaez Villarrubia, Claudia W; Stariha, Sarah; Babanova, Sofia; Schuler, Andrew J; Artyushkova, Kateryna; Atanassov, Plamen

    2015-03-01

    Non-Pt-group metal (non-PGM) materials based on transition metal-nitrogen-carbon (M-N-C) and derived from iron salt and aminoantipyrine (Fe-AAPyr) of mebendazole (Fe-MBZ) were studied for the first time as cathode catalysts in double-chamber microbial fuel cells (DCMFCs). The pH value of the cathode chamber was varied from 6 to 11 to elucidate the activity of those catalysts in acidic to basic conditions. The Fe-AAPyr- and Fe-MBZ-based cathodes were compared to a Pt-based cathode used as a baseline. Pt cathodes performed better at pH 6-7.5 and had similar performances at pH 9 and a substantially lower performance at pH 11 at which Fe-AAPyr and Fe-MBZ demonstrated their best electrocatalytic activity. The power density achieved with Pt constantly decreased from 94-99 μW cm(-2) at pH 6 to 55-57 μW cm(-2) at pH 11. In contrast, the power densities of DCMFs using Fe-AAPyr and Fe-MBZ were 61-68 μW cm(-2) at pH 6, decreased to 51-58 μW cm(-2) at pH 7.5, increased to 65-75 μW cm(-2) at pH 9, and the highest power density was achieved at pH 11 (68-80 μW cm(-2) ). Non-PGM cathode catalysts can be manufactured at the fraction of the cost of the Pt-based ones. The higher performance and lower cost indicates that non-PGM catalysts may be a viable materials choice in large-scale microbial fuel cells. PMID:25606716

  5. ATM Alters the Otherwise Robust Chromatin Mobility at Sites of DNA Double-Strand Breaks (DSBs) in Human Cells

    PubMed Central

    Becker, Annabelle; Durante, Marco; Taucher-Scholz, Gisela; Jakob, Burkhard

    2014-01-01

    Ionizing radiation induces DNA double strand breaks (DSBs) which can lead to the formation of chromosome rearrangements through error prone repair. In mammalian cells the positional stability of chromatin contributes to the maintenance of genome integrity. DSBs exhibit only a small, submicron scale diffusive mobility, but a slight increase in the mobility of chromatin domains by the induction of DSBs might influence repair fidelity and the formation of translocations. The radiation-induced local DNA decondensation in the vicinity of DSBs is one factor potentially enhancing the mobility of DSB-containing chromatin domains. Therefore in this study we focus on the influence of different chromatin modifying proteins, known to be activated by the DNA damage response, on the mobility of DSBs. IRIF (ionizing radiation induced foci) in U2OS cells stably expressing 53BP1-GFP were used as a surrogate marker of DSBs. Low angle charged particle irradiation, known to trigger a pronounced DNA decondensation, was used for the defined induction of linear tracks of IRIF. Our results show that movement of IRIF is independent of the investigated chromatin modifying proteins like ACF1 or PARP1 and PARG. Also depletion of proteins that tether DNA strands like MRE11 and cohesin did not alter IRIF dynamics significantly. Inhibition of ATM, a key component of DNA damage response signaling, resulted in a pronounced confinement of DSB mobility, which might be attributed to a diminished radiation induced decondensation. This confinement following ATM inhibition was confirmed using X-rays, proving that this effect is not restricted to densely ionizing radiation. In conclusion, repair sites of DSBs exhibit a limited mobility on a small spatial scale that is mainly unaffected by depletion of single remodeling or DNA tethering proteins. However, it relies on functional ATM kinase which is considered to influence the chromatin structure after irradiation. PMID:24651490

  6. Impact of Charged Particle Exposure on Homologous DNA Double-Strand Break Repair in Human Blood-Derived Cells

    PubMed Central

    Rall, Melanie; Kraft, Daniela; Volcic, Meta; Cucu, Aljona; Nasonova, Elena; Taucher-Scholz, Gisela; Bönig, Halvard; Wiesmüller, Lisa; Fournier, Claudia

    2015-01-01

    Ionizing radiation generates DNA double-strand breaks (DSB) which, unless faithfully repaired, can generate chromosomal rearrangements in hematopoietic stem and/or progenitor cells (HSPC), potentially priming the cells towards a leukemic phenotype. Using an enhanced green fluorescent protein (EGFP)-based reporter system, we recently identified differences in the removal of enzyme-mediated DSB in human HSPC versus mature peripheral blood lymphocytes (PBL), particularly regarding homologous DSB repair (HR). Assessment of chromosomal breaks via premature chromosome condensation or γH2AX foci indicated similar efficiency and kinetics of radiation-induced DSB formation and rejoining in PBL and HSPC. Prolonged persistence of chromosomal breaks was observed for higher LET charged particles which are known to induce more complex DNA damage compared to X-rays. Consistent with HR deficiency in HSPC observed in our previous study, we noticed here pronounced focal accumulation of 53BP1 after X-ray and carbon ion exposure (intermediate LET) in HSPC versus PBL. For higher LET, 53BP1 foci kinetics was similarly delayed in PBL and HSPC suggesting similar failure to repair complex DNA damage. Data obtained with plasmid reporter systems revealed a dose- and LET-dependent HR increase after X-ray, carbon ion and higher LET exposure, particularly in HR-proficient immortalized and primary lymphocytes, confirming preferential use of conservative HR in PBL for intermediate LET damage repair. HR measured adjacent to the leukemia-associated MLL breakpoint cluster sequence in reporter lines revealed dose dependency of potentially leukemogenic rearrangements underscoring the risk of leukemia-induction by radiation treatment. PMID:26618143

  7. Continuous Registration of Membrane Input Resistances of Small Plant Cells Using a Double-Pulse Current Clamp Technique for Single-Electrode Impalements 12

    PubMed Central

    Schefczik, Kurt; Simonis, Wilhelm; Schiebe, Michael

    1983-01-01

    To measure the cell input resistance in Elodea leaf cells, a new single-microelectrode method was explored by comparing the results with conventional two-microelectrode experiments. The new method takes advantage of the difference in the frequency response curves between electrode and cell impedances. By application of electrical stimuli, which contain specific frequency bands, the different impedances can be analyzed separately. To get a distinct separation in the frequency response of cell and electrode, respectively, the electrode capacitance has to be compensated during the impalement. Different time constants of the cell membrane can be accounted for by adjustment of the stimulus length. It is shown that both the single- and the double-electrode method yield the same results, even if the cell input resistances change considerably during the course of the experiment. This demonstrates the usefulness of the new single-electrode method for continuous measurements of cell membrane resistances, especially in cells so small that the double-electrode method is no longer applicable. Images Fig. 4 Fig. 2 PMID:16663008

  8. Double junction photoelectrochemical solar cells based on Cu2ZnSnS4/Cu2ZnSnSe4 thin film as composite photocathode

    NASA Astrophysics Data System (ADS)

    Zhu, L.; Qiang, Y. H.; Zhao, Y. L.; Gu, X. Q.

    2014-02-01

    A solvothermal method was used to synthesize Cu2ZnSnS4 (CZTS) and Cu2ZnSnSe4 (CZTSe) nanoparticles. CZTS/CZTSe bilayer films have been fabricated via a layer-by-layer blade coating process on the fluorine dope tin oxide (FTO) substrates. We converted conventional dye-sensitized solar cells (DSSCs) into double junction photoelectrochemical solar cells with the replacement of the Pt-coated counter electrode with the as-prepared films as composite photocathodes. Compared with conventional DSSCs, the cells show an increased short circuit current and power conversion efficiency.

  9. Cytotoxic data of 14-deoxy-11, 12-didehydroandrographolide (14-DDA), double transfection and DDIT3 silencing data in T-47D breast carcinoma cells.

    PubMed

    Tan, Heng Kean; Tengku Muhammad, Tengku Sifzizul; Tan, Mei Lan

    2016-06-01

    The data presented in this article are related to the research article entitled "14-deoxy-11,12-didehydroandrographolide induces DDIT3-dependent endoplasmic reticulum stress-mediated autophagy in T-47D breast carcinoma cells", which the mechanistic toxicology properties of 14-deoxy-11,12-didehydroandrographolide (14-DDA) were investigated (Tan et al., 2016 [1]). This article describes the derivation of cytotoxic parameters of 14-DDA, cell viability data after double transfection and DDIT3 silencing in T-47D cells. PMID:27182548

  10. A prospective randomized double-blind clinical trial using a combination of olfactory ensheathing cells and Schwann cells for the treatment of chronic complete spinal cord injuries.

    PubMed

    Chen, Lin; Huang, Hongyun; Xi, Haitao; Zhang, Feng; Liu, Yancheng; Chen, Di; Xiao, Juan

    2014-01-01

    The aim of this prospective randomized double-blind clinical study is to examine the benefits of using olfactory ensheathing cells (OECs) combined with or without Schwann cells (SCs) in treating chronic complete spinal cord injuries (SCIs). This would offer patients a better alternative for neurological functional recovery. According to the initial design, 28 eligible participants with cervical chronic complete SCI were recruited and randomly allocated into four groups of seven participants each. The neurological assessments were to be performed according to the American Spinal Injury Association (ASIA) and International Association of Neurorestoratology (IANR) Functional Rating Scales, in combination with electrophysiological tests, for example, electromyography (EMG) and paraspinal somatosensory evoked potentials (PSSEPs). Here we have summarized the data from seven patients; three patients received an OEC intraspinal transplantation, one underwent SC implantation, and one received a combination of OECs and SCs. The remaining two patients were used as controls. The scores were evaluated independently by at least two neurologists in a blinded fashion for comparing the neurological functional changes during pre- and post-cell transplantation (6-month follow-up). All patients who received OECs, SCs alone, and a combination of them showed functional improvement. Mild fever occurred in one of the patients with OEC transplant that subsided after symptomatic treatments. All treated patients except one showed improvement in the electrophysiological tests. The functional improvement rate comprises 5/5 (100%) in the treated group, but 0/2 (0%) in the control group (p = 0.008). These preliminary findings show that transplanting OECs, SCs, or a combination of them is well tolerated and that they have beneficial effects in patients. Thus, further studies in larger patient cohorts are warranted to assess the benefits and risks of these intervention strategies. This

  11. Double labelling of human umbilical cord mesenchymal stem cells with Gd-DTPA and PKH26 and the influence on biological characteristics of hUCMSCs.

    PubMed

    Shuai, Hanlin; Shi, Changzheng; Lan, Jifa; Chen, Danliang; Luo, Xin

    2015-02-01

    The aim of this study was to determine whether double labelling of human umbilical cord mesenchymal stem cells (hUCMSCs) with gadolinium-diethylene triamine penta-acetic acid (Gd-DTPA) and PKH26 influences their biological characteristics. A tissue adherence technique was used to separate and purify the hUCMSCs and flow cytometry was performed to detect the surface markers expressed on them. Gd-DTPA and PKH26 were used to label the stem cells and MRI and fluorescence microscopy were used to detect the double-labelled hUCMSCs. A MTT assay was used to delineate the growth curve. Transmission electron microscopy (TEM) and atomic force microscopy were used to demonstrate the ultrastructural features of the hUCMSCs. Flow cytometry showed that hUCMSCs highly expressed CD29, CD90, CD44 and CD105. No expression of CD31, CD34 and CD45 was detected. Very low expression of HLA-DR and CD40 was detected. Atomic force microscopy showed these cells were long, spindle shaped, and the cytoplasm and nucleus had clear boundaries. After double labelling, TEM showed Gd particles aggregated in the cytoplasm in a cluster pattern. The proliferation activity, cell cycle, apoptosis and differentiation of the stem cells were not influenced by double labelling. Thus a tissue adherence technique is helpful to separate and purify hUCMSCs effectively; and Gd-DTPA and PKH26 are promising tracers in the investigation of migration and distribution of hUCMSCs in vivo. PMID:25649907

  12. Survival of interneurons and parallel fiber synapses in a cerebellar cortex deprived of Purkinje cells: studies in the double mutant mouse Grid2Lc/+;Bax(-/-).

    PubMed

    Zanjani, S Hadi; Selimi, Fekrije; Vogel, Michael W; Haeberlé, Anne-Marie; Boeuf, Julien; Mariani, Jean; Bailly, Yannick J

    2006-08-01

    The Lurcher mutation in the Grid2 gene causes the cell autonomous death of virtually all cerebellar Purkinje cells and the target-related death of 90% of the granule cells and 60-75% of the olivary neurons. Inactivation of Bax, a pro-apoptotic gene of the Bcl-2 family, in heterozygous Lurcher mutants (Grid2Lc/+) rescues approximately 60% of the granule cells, but does not rescue Purkinje or olivary neurons. Given the larger size of the cerebellar molecular layer in Grid2Lc/+;Bax(-/-) double mutants compared to Grid2Lc/+ mutants, we analyzed the survival of the stellate and basket interneurons as well as the synaptic connectivity of parallel fibers originating from the surviving granule cells in the absence of their Purkinje cell targets in the Grid2Lc/+;Bax(-/-) cerebellum. Quantification showed a significantly higher density of interneurons ( approximately 60%) in the molecular layer of the Grid2Lc/+;Bax(-/-) mice compared to Grid2Lc/+, suggesting that interneurons are subject to a BAX-dependent target-related death in the Lurcher mutants. Furthermore, electron microscopy showed the normal ultrastructural aspect of a number of parallel fibers in the molecular layer of the Grid2Lc/+; Bax(-/-) double mutant mice and preserved their numerous synaptic contacts on interneurons, suggesting that interneurons could play a trophic role for axon terminals of surviving granule cells. Finally, parallel fibers varicosities in the double mutant established "pseudo-synapses" on glia as well as displayed autophagic profiles, suggesting that the connections established by the parallel fibers in the absence of their Purkinje cell targets were subject to a high turnover involving autophagy. PMID:16739195

  13. Defective selection of thymic regulatory T cells accompanies autoimmunity and pulmonary infiltrates in Tcra-deficient mice double transgenic for human La/SS-B and human La-specific T cell receptor1

    PubMed Central

    Yaciuk, Jane C.; Pan, Yujun; Schwarz, Karen; Pan, Zi-jian; Maier-Moore, Jacen S.; Kosanke, Stanley D.; Lawrence, Christina; Farris, A. Darise

    2014-01-01

    A human La/SS-B (hLa)-specific TCR/hLa neo-self antigen double transgenic mouse model was developed and used to investigate cellular tolerance and autoimmunity to the ubiquitous RNA-binding La antigen often targeted in systemic lupus erythematosus and Sjögren's syndrome. Extensive thymic clonal deletion of CD4+ T cells occurred in H-2k/k double transgenic mice presenting high levels of the I-Ek-restricted hLa T cell epitope. In contrast, deletion was less extensive in H-2k/b double transgenic mice presenting lower levels of the epitope, and some surviving thymocytes were positively selected as thymic regulatory T cells (tTreg). These mice remained serologically tolerant to hLa and healthy. H-2k/b double transgenic mice deficient of all endogenous Tcra genes, a deficiency known to impair Treg development and function, produced IgG anti-hLa autoantibodies and displayed defective tTreg development. These autoimmune mice had interstitial lung disease characterized by lymphocytic aggregates containing transgenic T cells with an activated, effector memory phenotype. Salivary gland infiltrates were notably absent. Thus, expression of nuclear hLa antigen induces thymic clonal deletion and tTreg selection, and lymphocytic infiltration of the lung is a consequence of La-specific CD4+ T cell autoimmunity. PMID:25582858

  14. Double Perovskite Anode Materials Sr2MMoO6 (M = Co, Ni) for Solid-Oxide Fuel Cells

    SciTech Connect

    Huang, Y.; Liang, G; Croft, M; Lehtimaki, M; Karppinen, M; Goodenough, J

    2009-01-01

    Double-perovskites Sr2MMoO6 (M = Co, Ni) have been investigated as anode materials for a solid oxide fuel cell. At room temperature, both Sr2CoMoO6 and Sr2NiMoO6 are tetragonal (I4/m). X-ray absorption spectroscopy confirmed the presence of Co2+/Mo6+ and Ni2+/Mo6+ pairs in the oxygen-stoichiometric compounds. The samples contain a limited concentration of oxygen vacancies in the reducing atmospheres at an anode. Reoxidation is facile below 600 C; they become antiferromagnetic at low temperatures TN = 37 and 80 K for M = Co and Ni, respectively. As an anode with a 300 em thick La0.8Sr0.2Ga0.83Mg0.17O2.815 electrolyte and SrFe0.2Co0.8O3-d as a cathode, Sr2CoMoO6 exhibited maximum power densities of 735 mW/cm2 in H2 and 527 mW/cm2 in wet CH4 at 800 C; Sr2NiMoO6 shows a notable power output only in dry CH4. The high performance of Sr2CoMoO6 in wet CH4 may be due to its catalytic effect on steam reforming of methane, but some degradation of the structure that occurred in CH4 obscures identification of the catalytic reaction processes at the surface. However, the stronger octahedral-site preference of Ni2+ versus Co2+ can account for the lower performance of the M = Ni anode.

  15. Population doubling level-dependent change of secreted glycosaminoglycan in equine bone marrow-derived mesenchymal stem cells

    PubMed Central

    SASAO, Takafumi; FUKUDA, Yuki; YOSHIDA, Sayako; MIYABARA, Shihori; KASASHIMA, Yoshinori; KUWANO, Atsutoshi; ARAI, Katsuhiko

    2015-01-01

    ABSTRACT In regenerative medicine using transplantation of mesenchymal stem cells (MSCs), the importance of regulating the quality of MSCs has been well recognized; however, there is little information concerning the relationship between the population doubling level (PDL) and the stemness of MSCs in equine medicine. In this study, we showed that the amount of glycosaminoglycan (GAG) secreted by bone marrow-derived MSCs (BMSCs) decreases with increase of PDL. Enzymatic digestion and two-dimensional electrophoresis revealed that a main component of GAG produced by BMSCs was hyaluronan with a small amount of chondroitin sulfate. Increase of PDL downregulated the expression of MSC CD markers, including CD44, CD73, CD90, CD105, and CD146, along with loss of differentiation capacity. Thus, the effect of hyaluronan supplement to the growth medium on both expression of CD markers and the tri-lineage potential of BMSCs was evaluated. Expression of CD73 and CD90 was preserved by continuous addition of hyaluronan to the growth medium, whereas mRNA levels corresponding to CD44, CD105 and CD146 were not preserved by supplementation of hyaluronan. BMSCs subcultured with hyaluronan-supplemented growth medium to PDL-12 showed osteogenic capacity, however adipogenic and chondrogenic activities at PDL-12 were not preserved by exogenous hyaluronan. These results suggest that downregulation of CD44, CD105 and CD146 might not affect the osteogenic capacity. Taken together, the results suggested that supplementation of hyaluronan to the growth medium might be effective at maintaining the osteogenic capacity of equine BMSCs. PMID:26435680

  16. Asymmetric segregation of the double-stranded RNA binding protein Staufen2 during mammalian neural stem cell divisions promotes lineage progression.

    PubMed

    Kusek, Gretchen; Campbell, Melissa; Doyle, Frank; Tenenbaum, Scott A; Kiebler, Michael; Temple, Sally

    2012-10-01

    Asymmetric cell divisions are a fundamental feature of neural development, and misregulation can lead to brain abnormalities or tumor formation. During an asymmetric cell division, molecular determinants are segregated preferentially into one daughter cell to specify its fate. An important goal is to identify the asymmetric determinants in neural progenitor cells, which could be tumor suppressors or inducers of specific neural fates. Here, we show that the double-stranded RNA-binding protein Stau2 is distributed asymmetrically during progenitor divisions in the developing mouse cortex, preferentially segregating into the Tbr2(+) neuroblast daughter, taking with it a subset of RNAs. Knockdown of Stau2 stimulates differentiation and overexpression produces periventricular neuronal masses, demonstrating its functional importance for normal cortical development. We immunoprecipitated Stau2 to examine its cargo mRNAs, and found enrichment for known asymmetric and basal cell determinants, such as Trim32, and identified candidates, including a subset involved in primary cilium function. PMID:22902295

  17. Cytotoxic data of 14-deoxy-11, 12-didehydroandrographolide (14-DDA), double transfection and DDIT3 silencing data in T-47D breast carcinoma cells

    PubMed Central

    Tan, Heng Kean; Tengku Muhammad, Tengku Sifzizul; Tan, Mei Lan

    2016-01-01

    The data presented in this article are related to the research article entitled “14-deoxy-11,12-didehydroandrographolide induces DDIT3-dependent endoplasmic reticulum stress-mediated autophagy in T-47D breast carcinoma cells”, which the mechanistic toxicology properties of 14-deoxy-11,12-didehydroandrographolide (14-DDA) were investigated (Tan et al., 2016 [1]). This article describes the derivation of cytotoxic parameters of 14-DDA, cell viability data after double transfection and DDIT3 silencing in T-47D cells. PMID:27182548

  18. Dynamics and Cell-Type Specificity of the DNA Double-Strand Break Repair Protein RecN in the Developmental Cyanobacterium Anabaena sp. Strain PCC 7120

    PubMed Central

    Hu, Sheng; Wang, Jinglan; Wang, Li; Zhang, Cheng-Cai; Chen, Wen-Li

    2015-01-01

    DNA replication and repair are two fundamental processes required in life proliferation and cellular defense and some common proteins are involved in both processes. The filamentous cyanobacterium Anabaena sp. strain PCC 7120 is capable of forming heterocysts for N2 fixation in the absence of a combined-nitrogen source. This developmental process is intimately linked to cell cycle control. In this study, we investigated the localization of the DNA double-strand break repair protein RecN during key cellular events, such as chromosome damaging, cell division, and heterocyst differentiation. Treatment by a drug causing DNA double-strand breaks (DSBs) induced reorganization of the RecN focus preferentially towards the mid-cell position. RecN-GFP was absent in most mature heterocysts. Furthermore, our results showed that HetR, a central player in heterocyst development, was involved in the proper positioning and distribution of RecN-GFP. These results showed the dynamics of RecN in DSB repair and suggested a differential regulation of DNA DSB repair in vegetative cell and heterocysts. The absence of RecN in mature heterocysts is compatible with the terminal nature of these cells. PMID:26431054

  19. Differential repair of radiation-induced DNA damage in cells of human squamous cell carcinoma and the effect of caffeine and cysteamine on induction and repair of DNA double-strand breaks

    SciTech Connect

    Smeets, M.F.M.A.; Mooren, E.H.M.; Abdel-Wahab, A.H.A.; Begg, A.C.

    1994-11-01

    The goal of these experiments was to investigate further the relationship between DNA double-strand breaks and cell killing in human tumor cells, first by comparing different cell lines, and second by radiomodification studies. Field-inversion gel electrophoresis was used to quantify double-strand breaks. Two subclones of the radioresistant human squamous cell carcinoma line SQ20B (SQD9 and SQG6) were compared. These subclones differed in DNA index by a factor of 1.7 but showed the same resistance to radiation as cells of the parental cell line. It was found that, although induction of DSBs was not significantly different in the two cell lines, the t{sub 1/2} of the fast component of repair was significantly shorter for SQD9 cells, leading to greater overall repair which was not reflected in increased survival. Caffeine and cysteamine were tested as modifiers of radiosensitivity, using the radioresistant SQ20B line and the radiosensitive SCC61 cell line. No effect of caffeine was seen when the drug was present only during irradiation. Postirradiation incubations with caffeine, however, resulted in a dose reduction factor greater than 2.0 in cell survival for both cell lines. In contrast, induction of DSBs was reduced by caffeine, and no effect on DSB repair was observed. Cysteamine led to a dose protection factor greater than 1.8 in cell survival in both cell lines. A reduction in induced DSBs was found at high doses corresponding approximately with the increase in cell survival. Over the same (low) dose range, however, the correlation between DSB induction and cell killing was poor. These data indicate that DSB induction does not correlate well with cell killing either for different cell lines, for radiochemical modification (cysteamine) or for some other types of modification (caffeine). 31 refs., 8 figs.

  20. Variable content of double minute chromosomes is not correlated with degree of phenotype instability in methotrexate-resistant human cell lines

    SciTech Connect

    Masters, J.; Keeley, B.; Gay, H.; Attardi, G.

    1982-05-01

    Several variants resistant to 1.8 x 10/sup -4/ M DL-methotrexate (MTX) have been isolated from the human cell lines HeLa BU25 and VA/sub 2/-B by exposing them to progressively increasing concentrations of the drug. A striking variability of phenotype and chromosome constitution was observed among the different variants. All resistant cell lines exhibited a greatly increased dihydrofolic acid reductase (DHFR) activity and DHFR content; however, the DHFR activity levels varied considerably among the variants, ranging between about 35 and 275 times the parental level. An analysis of the correlaion of loss of double minute chromosomes and loss of DHFR activity in the absence of MTX has given results consistent with the idea that the double-minute chromosomes contain amplified DHFR genes. However, the most significant finding is that, in contrast to what has been reported in the mouse system, the recognizable double-minute chromosomes varied greatly in number in different variants without any relationship to either the level of DHFR activity or the degree of instability of MTX resistance in the absence of selective pressure. These and other observations point to the occurrence in the human MTX-resistant variants of another set of DHFR genes, representing a varied proportion of the total, which is associated with the regular chromosomes, and which may be unstable in the absence of selective pressure.

  1. The effect of 2-[(aminopropyl)amino] ethanethiol (WR-1065) on radiation induced DNA double strand damage and repair in V79 cells.

    PubMed Central

    Sigdestad, C. P.; Treacy, S. H.; Knapp, L. A.; Grdina, D. J.

    1987-01-01

    Radiation induced DNA double strand breaks are believed to be important lesions involved in processes related to cell killing, induction of chromosome aberrations and carcinogenesis. This paper reports the effects of the radioprotector 2-[(aminopropyl)amino]ethanethiol (WR-1065) on radiation-induced DNA damage and repair in V79 cells using the neutral elution method performed at pH 7.2 or pH 9.6. WR-1065 (4 mM) was added to the culture medium either 30 minutes prior to and during irradiation with Cobalt-60 gamma rays (for dose response experiments) or during the repair times tested (for DNA rejoining experiments). The results indicate that WR-1065 is an effective protector against the formation of radiation-induced double-strand breaks in DNA as measured using a neutral elution technique at either pH. The protector reduced the strand scission factors by 1.44 and 1.77 in experiments run at pH 9.6 and pH 7.2, respectively. The kinetics of DNA double-strand rejoining were dependent upon the pH at which the neutral elution procedure was performed. Unlike the results obtained with alkaline elution, rejoining of DNA breaks was unaffected by the presence of WR-1065 at either pH. PMID:3606941

  2. Development of a Split SNAP-CLIP Double Labeling System for Tracking Proteins Following Dissociation from Protein-Protein Complexes in Living Cells.

    PubMed

    Mie, Masayasu; Naoki, Tatsuhiko; Kobatake, Eiry

    2016-08-16

    The split SNAP-tag protein-fragment complementation assay (PCA) is a useful tool for imaging protein-protein interactions (PPIs) in living cells. In contrast to conventional methods employed for imaging PPIs, the split SNAP-tag PCA enables tracking of proteins following dissociation from protein-protein complexes. A limitation of this system, however, is that it only allows for labeling and tracking of one of the proteins forming the protein-protein complex. To track both proteins forming a protein-protein complex, each protein needs to be appropriately labeled. In this study, a split SNAP-CLIP double labeling system is developed and applied for tracking of each protein forming a protein-protein complex. As a proof-of concept, FM protein for PPIs and protein kinase C alpha (PKCα) for translocation are introduced to a split SNAP-CLIP double labeling system. The results show a split SNAP-CLIP double labeling system enables labeling of both proteins in a protein-protein complex and subsequent tracking of each of the proteins following dissociation from the protein-protein complexes in living cells. PMID:27448142

  3. IL-9 promotes the survival and function of human melanoma-infiltrating CD4(+) CD8(+) double-positive T cells.

    PubMed

    Parrot, Tiphaine; Allard, Mathilde; Oger, Romain; Benlalam, Houssem; Raingeard de la Blétière, Diane; Coutolleau, Anne; Preisser, Laurence; Desfrançois, Juliette; Khammari, Amir; Dréno, Brigitte; Labarrière, Nathalie; Delneste, Yves; Guardiola, Philippe; Gervois, Nadine

    2016-07-01

    We previously demonstrated an accumulation of tumor-reactive CD4(+) CD8(+) double positive (DP) T cells within melanoma-infiltrating lymphocytes, supporting their role in the regulation of anti-tumor immune responses. Similarly to their CD8(+) counterparts, intra-tumor DP T cells are MHC class-I restricted but differed by a limited lytic activity against autologous melanoma cells. Based on these observations and to further characterize DP T cells, both populations were compared at the transcriptional level. Our results revealed the overexpression of the IL-9 receptor (IL-9R) by DP T cells and prompted us to investigate the impact of IL-9 on their biology. We show that IL-9 favors DP T-cell survival by protecting them from apoptosis and by promoting their proliferation. In addition, IL-9 enhances their ability to produce cytokines and increased their levels of granzyme B/perforin as well as degranulation capacity, leading to a strengthened cytotoxic activity against melanoma cells. Taken together, the IL-9R(high) DP T-cell population could be a new preferential target for IL-9, which could take part in their retention within the melanoma infiltrate while also favoring their anti-tumor activity. More generally, our results extend the pleiotropic effects of IL-9 to IL-9R-expressing intra-tumor T cells, which could further potentiate anti-tumor immune responses. PMID:27094152

  4. The production of alpha/beta and gamma/delta double negative (DN) T-cells and their role in the maintenance of pregnancy.

    PubMed

    Chapman, John C; Chapman, Fae M; Michael, Sandra D

    2015-01-01

    The ability of the thymus gland to convert bone marrow-derived progenitor cells into single positive (SP) T-cells is well known. In this review we present evidence that the thymus, in addition to producing SP T-cells, also has a pathway for the production of double negative (DN) T-cells. The existence of this pathway was noted during our examination of relevant literature to determine the cause of sex steroid-induced thymocyte loss. In conducting this search our objective was to answer the question of whether thymocyte loss is the end product of a typical interaction between the reproductive and immune systems, or evidence that the two systems are incompatible. We can now report that "thymocyte loss" is a normal process that occurs during the production of DN T-cells. The DN T-cell pathway is unique in that it is mediated by thymic mast cells, and becomes functional following puberty. Sex steroids initiate the development of the pathway by binding to an estrogen receptor alpha located in the outer membrane of the mast cells, causing their activation. This results in their uptake of extracellular calcium, and the production and subsequent release of histamine and serotonin. Lymphatic vessels, located in the subcapsular region of the thymus, respond to the two vasodilators by undergoing a substantial and preferential uptake of gamma/delta and alpha/beta DN T- cells. These T- cells exit the thymus via efferent lymphatic vessels and enter the lymphatic system.The DN pathway is responsible for the production of three subsets of gamma/delta DN T-cells and one subset of alpha/beta DN T-cells. In postpubertal animals approximately 35 % of total thymocytes exit the thymus as DN T-cells, regardless of sex. In pregnant females, their levels undergo a dramatic increase. Gamma/delta DN T-cells produce cytokines that are essential for the maintenance of pregnancy. PMID:26164866

  5. Double Emulsion Templated Celloidosomes

    NASA Astrophysics Data System (ADS)

    Arriaga, Laura R.; Marquez, Samantha M.; Kim, Shin-Hyun; Chang, Connie; Wilking, Jim; Monroy, Francisco; Marquez, Manuel; Weitz, David A.

    2012-02-01

    We present a novel approach for fabricating celloidosomes, which represent a hollow and spherical three-dimensional self-assembly of living cells encapsulating an aqueous core. Glass- capillary microfluidics is used to generate monodisperse water-in-oil-in-water double emulsion templates using lipids as stabilizers. Such templates allow for obtaining single but also double concentric celloidosomes. In addition, after a solvent removal step the double emulsion templates turn into monodisperse lipid vesicles, whose membrane spontaneously phase separates when choosing the adequate lipid composition, providing the adequate scaffold for fabricating Janus-celloidosomes. These structures may find applications in the development of bioreactors in which the synergistic effects of two different types of cells selectively adsorbed on one of the vesicle hemispheres may be exploited.

  6. Double isotopic method using dansyl chloride for the determination of GABA in rat C6 astrocytoma cell cultures

    SciTech Connect

    Kohl, R.L.; Quay, W.B.; Perez-Polo, J.R.

    1986-01-01

    Methods are described for the quantitative measurement of GABA in culture. The method can be adapted to any amino acid or dansyl-chloride-reactive species. The sensitivity and selectivity of the procedure result from the double isotopic design in which (/sup 14/C)-labeled internal standard was added to the samples before reaction with (3M)-labeled dansyl chloride. Values obtained by ion-exchange amino acid analysis of cultures agree closely with the values obtained by the double isotopic method. This method is sensitive enough to measure GABA intracellularly and the condition medium.

  7. Oilseed rape seeds with ablated defence cells of the glucosinolate–myrosinase system. Production and characteristics of double haploid MINELESS plants of Brassica napus L.

    PubMed Central

    Ahuja, Ishita; Borgen, Birgit Hafeld; Hansen, Magnor; Honne, Bjørn Ivar; Müller, Caroline; Rohloff, Jens; Rossiter, John Trevor; Bones, Atle Magnar

    2011-01-01

    Oilseed rape and other crop plants of the family Brassicaceae contain a unique defence system known as the glucosinolate–myrosinase system or the ‘mustard oil bomb’. The ‘mustard oil bomb’ which includes myrosinase and glucosinolates is triggered by abiotic and biotic stress, resulting in the formation of toxic products such as nitriles and isothiocyanates. Myrosinase is present in specialist cells known as ‘myrosin cells’ and can also be known as toxic mines. The myrosin cell idioblasts of Brassica napus were genetically reprogrammed to undergo controlled cell death (ablation) during seed development. These myrosin cell-free plants have been named MINELESS as they lack toxic mines. This has led to the production of oilseed rape with a significant reduction both in myrosinase levels and in the hydrolysis of glucosinolates. Even though the myrosinase activity in MINELESS was very low compared with the wild type, variation was observed. This variability was overcome by producing homozygous seeds. A microspore culture technique involving non-fertile haploid MINELESS plants was developed and these plants were treated with colchicine to produce double haploid MINELESS plants with full fertility. Double haploid MINELESS plants had significantly reduced myrosinase levels and glucosinolate hydrolysis products. Wild-type and MINELESS plants exhibited significant differences in growth parameters such as plant height, leaf traits, matter accumulation, and yield parameters. The growth and developmental pattern of MINELESS plants was relatively slow compared with the wild type. The characteristics of the pure double haploid MINELESS plant are described and its importance for future biochemical, agricultural, dietary, functional genomics, and plant defence studies is discussed. PMID:21778185

  8. Overexpression of the scaffold WD40 protein WRAP53β enhances the repair of and cell survival from DNA double-strand breaks.

    PubMed

    Rassoolzadeh, H; Böhm, S; Hedström, E; Gad, H; Helleday, T; Henriksson, S; Farnebo, M

    2016-01-01

    Altered expression of the multifunctional protein WRAP53β (WD40 encoding RNA Antisense to p53), which targets repair factors to DNA double-strand breaks and factors involved in telomere elongation to Cajal bodies, is linked to carcinogenesis. While loss of WRAP53β function has been shown to disrupt processes regulated by this protein, the consequences of its overexpression remain unclear. Here we demonstrate that overexpression of WRAP53β disrupts the formation of and impairs the localization of coilin to Cajal bodies. At the same time, the function of this protein in the repair of DNA double-strand breaks is enhanced. Following irradiation, cells overexpressing WRAP53β exhibit more rapid clearance of phospho-histone H2AX (γH2AX), and more efficient homologous recombination and non-homologous end-joining, in association with fewer DNA breaks. Moreover, in these cells the ubiquitylation of damaged chromatin, which is known to facilitate the recruitment of repair factors and subsequent repair, is elevated. Knockdown of the ubiquitin ligase involved, ring-finger protein 8 (RNF8), which is recruited to DNA breaks by WRAP53β, attenuated this effect, suggesting that overexpression of WRAP53β leads to more rapid repair, as well as improved cell survival, by enhancing RNF8-mediated ubiquitylation at DNA breaks. Our present findings indicate that WRAP53β and RNF8 are rate-limiting factors in the repair of DNA double-strand breaks and raise the possibility that upregulation of WRAP53β may contribute to genomic stability in and survival of cancer cells. PMID:27310875

  9. Design friendly double patterning

    NASA Astrophysics Data System (ADS)

    Yesilada, Emek

    2012-03-01

    Double patterning using 193nm immersion has been adapted as the solution to enable 2x nm technology nodes until the arrival of EUV tools. As a result the past few years have seen a huge effort in creating double patterning friendly design flows. These flows have so far proposed a combination of decomposition rules at cell level and/or at placement level as well as sophisticated decomposition tools with varying density, design iteration and decomposition complexity penalties. What is more, designers have to familiarize themselves with double patterning challenges and decomposition tools. In this paper an alternative approach is presented that allows the development of dense standard cells with minimal impact on design flow due to double patterning. A real case study is done on 20nm node first metal layer where standard cells are designed without considering decomposition restrictions. The resulting layout is carefully studied in order to establish decomposition or color rules that can map the layout into two masks required for double patterning but without the need of complex coloring algorithms. Since the rules are derived from a decomposition unaware design they do not in return impose heavy restrictions on the design at the cell or placement level and show substantial density gains compared to previously proposed methods. Other key advantages are a simplified design flow without complex decomposition tools that can generate a faster time to market solution all at the same time keeping designers isolated from the challenges of the double patterning. The derived design rules highlight process development path required for design driven manufacturing.

  10. Effect of Broccoli Sprouts and Live Attenuated Influenza Virus on Peripheral Blood Natural Killer Cells: A Randomized, Double-Blind Study

    PubMed Central

    Müller, Loretta; Meyer, Megan; Bauer, Rebecca N.; Zhou, Haibo; Zhang, Hongtao; Jones, Shannon; Robinette, Carole; Noah, Terry L.; Jaspers, Ilona

    2016-01-01

    Enhancing antiviral host defense responses through nutritional supplementation would be an attractive strategy in the fight against influenza. Using inoculation with live attenuated influenza virus (LAIV) as an infection model, we have recently shown that ingestion of sulforaphane-containing broccoli sprout homogenates (BSH) reduces markers of viral load in the nose. To investigate the systemic effects of short-term BSH supplementation in the context of LAIV-inoculation, we examined peripheral blood immune cell populations in non-smoking subjects from this study, with a particular focus on NK cells. We carried out a randomized, double-blinded, placebo-controlled study measuring the effects of BSH (N = 13) or placebo (alfalfa sprout homogenate, ASH; N = 16) on peripheral blood mononuclear cell responses to a standard nasal vaccine dose of LAIV in healthy volunteers. Blood was drawn prior to (day-1) and post (day2, day21) LAIV inoculation and analyzed for neutrophils, monocytes, macrophages, T cells, NKT cells, and NK cells. In addition, NK cells were enriched, stimulated, and assessed for surface markers, intracellular markers, and cytotoxic potential by flow cytometry. Overall, LAIV significantly reduced NKT (day2 and day21) and T cell (day2) populations. LAIV decreased NK cell CD56 and CD158b expression, while significantly increasing CD16 expression and cytotoxic potential (on day2). BSH supplementation further increased LAIV-induced granzyme B production (day2) in NK cells compared to ASH and in the BSH group granzyme B levels appeared to be negatively associated with influenza RNA levels in nasal lavage fluid cells. We conclude that nasal influenza infection may induce complex changes in peripheral blood NK cell activation, and that BSH increases virus-induced peripheral blood NK cell granzyme B production, an effect that may be important for enhanced antiviral defense responses. Trial Registration: ClinicalTrials.gov NCT01269723 PMID:26820305

  11. Cyclin D1 silencing suppresses tumorigenicity, impairs DNA double strand break repair and thus radiosensitizes androgen-independent prostate cancer cells to DNA damage

    PubMed Central

    Ju, Xiaoming; Vetuschi, Antonella; Sferra, Roberta; Casimiro, Mathew C.; Pompili, Simona; Festuccia, Claudio; Colapietro, Alessandro; Gaudio, Eugenio; Di Cesare, Ernesto; Tombolini, Vincenzo; Pestell, Richard G.

    2016-01-01

    Patients with hormone-resistant prostate cancer (PCa) have higher biochemical failure rates following radiation therapy (RT). Cyclin D1 deregulated expression in PCa is associated with a more aggressive disease: however its role in radioresistance has not been determined. Cyclin D1 levels in the androgen-independent PC3 and 22Rv1 PCa cells were stably inhibited by infecting with cyclin D1-shRNA. Tumorigenicity and radiosensitivity were investigated using in vitro and in vivo experimental assays. Cyclin D1 silencing interfered with PCa oncogenic phenotype by inducing growth arrest in the G1 phase of cell cycle and reducing soft agar colony formation, migration, invasion in vitro and tumor formation and neo-angiogenesis in vivo. Depletion of cyclin D1 significantly radiosensitizes PCa cells by increasing the RT-induced DNA damages by affecting the NHEJ and HR pathways responsible of the DNA double-strand break repair. Following treatment of cells with RT the abundance of a biomarker of DNA damage, γ-H2AX, was dramatically increased in sh-cyclin D1 treated cells compared to shRNA control. Concordant with these observations DNA-PKcs-activation and RAD51-accumulation, part of the DNA double-strand break repair machinery, were reduced in shRNA-cyclin D1 treated cells compared to shRNA control. We further demonstrate the physical interaction between CCND1 with activated-ATM, -DNA-PKcs and RAD51 is enhanced by RT. Finally, siRNA-mediated silencing experiments indicated DNA-PKcs and RAD51 are downstream targets of CCND1-mediated PCa cells radioresistance. In summary, these observations suggest that CCND1 is a key mediator of PCa radioresistance and could represent a potential target for radioresistant hormone-resistant PCa. PMID:26689991

  12. Characterization of a novel NKG2D and NKp46 double-mutant mouse reveals subtle variations in the NK cell repertoire

    PubMed Central

    Sheppard, Sam; Triulzi, Chiara; Ardolino, Michele; Serna, Daniel; Zhang, Lily; Raulet, David H.

    2013-01-01

    The immunoreceptors NKG2D and NKp46 are known for their capacity to activate natural killer (NK) cell cytotoxicity and secretory responses in the contexts of tumors and infections, yet their roles in NK cell education remain unclear. Here, we provide the first characterization of mice deficient for both NKG2D and NKp46 receptors to address the relevance of their concomitant absence during NK cell development and function. Our findings reveal that NK cells develop normally in double-mutant (DKO) mice. Mice lacking NKG2D but not NKp46 showed subtle differences in the percentages of NK cells expressing inhibitory Ly49 receptors and the adhesion molecule DNAM-1. A slightly increased percentage of terminally differentiated NK cells and functional response to in vitro stimuli was observed in some experiments. These alterations were modest and did not affect NK cell function in vivo in response to mouse cytomegalovirus infection. NKp46 deficiency alone, or in combination with NKG2D deficiency, had no effect on frequency or function of NK cells. PMID:23649470

  13. Gene conversion is strongly induced in human cells by double-strand breaks and is modulated by the expression of BCL-x(L)

    NASA Technical Reports Server (NTRS)

    Wiese, Claudia; Pierce, Andrew J.; Gauny, Stacey S.; Jasin, Maria; Kronenberg, Amy; Chatterjee, A. (Principal Investigator)

    2002-01-01

    Homology-directed repair (HDR) of DNA double-strand breaks (DSBs) contributes to the maintenance of genomic stability in rodent cells, and it has been assumed that HDR is of similar importance in DSB repair in human cells. However, some outcomes of homologous recombination can be deleterious, suggesting that factors exist to regulate HDR. We demonstrated previously that overexpression of BCL-2 or BCL-x(L) enhanced the frequency of X-ray-induced TK1 mutations, including loss of heterozygosity events presumed to arise by mitotic recombination. The present study was designed to test whether HDR is a prominent DSB repair pathway in human cells and to determine whether ectopic expression of BCL-x(L) affects HDR. Using TK6-neo cells, we find that a single DSB in an integrated HDR reporter stimulates gene conversion 40-50-fold, demonstrating efficient DSB repair by gene conversion in human cells. Significantly, DSB-induced gene conversion events are 3-4-fold more frequent in TK6 cells that stably overexpress the antiapoptotic protein BCL-X(L). Thus, HDR plays an important role in maintaining genomic integrity in human cells, and ectopic expression of BCL-x(L) enhances HDR of DSBs. This is the first study to highlight a function for BCL-x(L) in modulating DSB repair in human cells.

  14. Environmentally relevant concentrations of arsenite and monomethylarsonous acid inhibit IL-7/STAT5 cytokine signaling pathways in mouse CD3+CD4-CD8- double negative thymus cells.

    PubMed

    Xu, Huan; Lauer, Fredine T; Liu, Ke Jian; Hudson, Laurie G; Burchiel, Scott W

    2016-04-15

    Environmental arsenic exposure is a public health issue. Immunotoxicity induced by arsenic has been reported in humans and animal models. The purpose of this study was to evaluate mechanisms of As(+3) and MMA(+3) toxicity in mouse thymus cells. Because we know that MMA(+3) inhibits IL-7 signaling in mouse bone marrow pre-B cells, we studied the influence of As(+3) and MMA(+3) on T cell development in the thymus at the earliest stage of T cell development (CD4-CD8-, double negative, DN) which requires IL-7 dependent signaling. We found in a DN thymus cell line (D1) that a low concentration of MMA(+3) (50 nM) suppressed IL-7 dependent JAK1, 3 and STAT5 signaling. As(+3) suppressed STAT5 and JAK3 at higher concentrations (500 nM). Cell surface expression of the IL-7 receptor (CD127) was also suppressed by 50 nM MMA(+)3, but was increased by 500 NM As(+3), indicating possible differences in the mechanisms of action of these agents. A decrease in cyclin D1 protein expression was observed in D1 cells exposed to As(+3) at 500 nM and MMA(+3) starting at 50 nM, suggesting that arsenic at these environmentally-relevant doses suppresses early T cell development through the inhibition of IL-7 signaling pathway. PMID:26921788

  15. Simultaneous visualization of extrinsic and intrinsic axon collaterals in Golgi-like detail for mouse corticothalamic and corticocortical cells: a double viral infection method

    PubMed Central

    Watakabe, Akiya; Takaji, Masafumi; Kato, Shigeki; Kobayashi, Kazuto; Mizukami, Hiroaki; Ozawa, Keiya; Ohsawa, Sonoko; Matsui, Ryosuke; Watanabe, Dai; Yamamori, Tetsuo

    2014-01-01

    Here we present a novel tracing technique to stain projection neurons in Golgi-like detail by double viral infection. We used retrograde lentiviral vectors and adeno-associated viral vectors (AAV) to drive “TET-ON/TET-OFF system” in neurons connecting two regions. Using this method, we successfully labeled the corticothalamic (CT) cells of the mouse somatosensory barrel field (S1BF) and motor cortex (M1) in their entirety. We also labeled contra- and ipsilaterally-projecting corticocortical (CC) cells of M1 by targeting contralateral M1 or ipsilateral S1 for retrograde infection. The strength of this method is that we can observe the morphology of specific projection neuron subtypes en masse. We found that the group of CT cells extends their dendrites and intrinsic axons extensively below but not within the thalamorecipient layer in both S1BF and M1, suggesting that the primary target of this cell type is not layer 4. We also found that both ipsi- and contralateral targeting CC cells in M1 commonly exhibit widespread collateral extensions to contralateral M1 (layers 1–6), bilateral S1 and S2 (layers 1, 5 and 6), perirhinal cortex (layers 1, 2/3, 5, and 6), striatum and claustrum. These findings not only strengthened the previous findings of single cell tracings but also extended them by enabling cross-area comparison of CT cells or comparison of CC cells of two different labeling. PMID:25278843

  16. Nontoxic concentration of DNA-PK inhibitor NU7441 radio-sensitizes lung tumor cells with little effect on double strand break repair.

    PubMed

    Sunada, Shigeaki; Kanai, Hideki; Lee, Younghyun; Yasuda, Takeshi; Hirakawa, Hirokazu; Liu, Cuihua; Fujimori, Akira; Uesaka, Mitsuru; Okayasu, Ryuichi

    2016-09-01

    High-linear energy transfer (LET) heavy ions have been increasingly employed as a useful alternative to conventional photon radiotherapy. As recent studies suggested that high LET radiation mainly affects the nonhomologous end-joining (NHEJ) pathway of DNA double strand break (DSB) repair, we further investigated this concept by evaluating the combined effect of an NHEJ inhibitor (NU7441) at a non-toxic concentration and carbon ions. NU7441-treated non-small cell lung cancer (NSCLC) A549 and H1299 cells were irradiated with X-rays and carbon ions (290 MeV/n, 50 keV/μm). Cell survival was measured by clonogenic assay. DNA DSB repair, cell cycle distribution, DNA fragmentation and cellular senescence induction were studied using a flow cytometer. Senescence-associated protein p21 was detected by western blotting. In the present study, 0.3 μM of NU7441, nontoxic to both normal and tumor cells, caused a significant radio-sensitization in tumor cells exposed to X-rays and carbon ions. This concentration did not seem to cause inhibition of DNA DSB repair but induced a significant G2/M arrest, which was particularly emphasized in p53-null H1299 cells treated with NU7441 and carbon ions. In addition, the combined treatment induced more DNA fragmentation and a higher degree of senescence in H1299 cells than in A549 cells, indicating that DNA-PK inhibitor contributes to various modes of cell death in a p53-dependent manner. In summary, NSCLC cells irradiated with carbon ions were radio-sensitized by a low concentration of DNA-PK inhibitor NU7441 through a strong G2/M cell cycle arrest. Our findings may contribute to further effective radiotherapy using heavy ions. PMID:27341700

  17. Artemis deficiency confers a DNA double-strand break repair defect and Artemis phosphorylation status is altered by DNA damage and cell cycle progression.

    PubMed

    Wang, Junhua; Pluth, Janice M; Cooper, Priscilla K; Cowan, Morton J; Chen, David J; Yannone, Steven M

    2005-05-01

    Mutations in the Artemis gene are causative in a subset of human severe combined immunodeficiencies (SCIDs) and Artemis-deficient cells exhibit radiation sensitivity and defective V(D)J recombination, implicating Artemis function in non-homologous end joining (NHEJ). Here we show that Artemis-deficient cells from Athabascan-speaking Native American SCID patients (SCIDA) display significantly elevated sensitivity to ionizing radiation (IR) but only a very subtle defect in DNA double-strand (DSB) break repair in contrast to the severe DSB repair defect of NHEJ-deficient cells. Primary human SCIDA fibroblasts accumulate and exhibit persistent arrest at both the G1/S and G2/M boundaries in response to IR, consistent with the presence of persistent DNA damage. Artemis protein is phosphorylated in a PI3-like kinase-dependent manner after either IR or a number of other DNA damaging treatments including etoposide, but SCIDA cells are not hypersensitive to treatment with etoposide. Inhibitor studies with various DNA damaging agents establish multiple phosphorylation states and suggest multiple kinases function in Artemis phosphorylation. We observe that Artemis phosphorylation occurs rapidly after irradiation like that of histone H2AX. However, unlike H2AX, Artemis de-phosphorylation is uncoupled from overall DNA repair and correlates instead with cell cycle progression to or through mitosis. Our results implicate a direct and non-redundant function of Artemis in the repair of a small subset of DNA double-strand breaks, possibly those with hairpin termini, which may account for the pronounced radiation sensitivity observed in Artemis-deficient cells. PMID:15811628

  18. A novel double carbonyl analog of curcumin induces the apoptosis of human lung cancer H460 cells via the activation of the endoplasmic reticulum stress signaling pathway.

    PubMed

    Ye, Hui; Wei, Xiaoyan; Wang, Zhankun; Zhang, Shanshan; Ren, Jiye; Yao, Song; Shi, Lingyi; Yang, Lizhu; Qiu, Peihong; Wu, Jianzhang; Liang, Guang

    2016-09-01

    Curcumin can inhibit the growth of a variety of cancer cells; however, its poor bioavailability and pharmacokinetic profiles, which are attributed to its instability under physiological conditions, have limited its application in anticancer therapy. In the present study, we screened a double carbonyl analog of curcumin (A17) and analyzed its effects and mechanism of inducing apoptosis in human lung cancer H460 cells. The results showed that A17 not only induced CHOP expression in human lung cancer H460 cells, but also induced the apoptosis of H460 cells in a dose-responsive manner, and this effect was related to corresponding activation of some important components in the endoplasmic reticulum (ER) stress-mediated apoptosis pathway. When CHOP was knocked down by specific siRNA, A17-induced cell apoptosis was attenuated, thereby further demonstrating that the apoptotic pathway is ER stress‑dependent. Our studies demonstrated that A17 has better stability and antitumor activity than curcumin in H460 cells via an ER stress-mediated mechanism. These results imply that A17 could be further explored as a potential anticancer agent for the treatment of human non-small cell lung cancer (NSCLC). PMID:27431486

  19. Double-layer electrode based on TiO2 nanotubes arrays for enhancing photovoltaic properties in dye-sensitized solar cells.

    PubMed

    He, Zuoli; Que, Wenxiu; Sun, Peng; Ren, Jiangbo

    2013-12-26

    The present work reports a rapid and facile method to fabricate a novel double-layer TiO2 photoanode, which is based on highly ordered TiO2 nanotube arrays and monodispersive scattering microspheres. This double-layer TiO2 sphere/TNTA photoanode have got many unique structural and optical properties from TiO2 scattering microspheres, such as high specific surface area, multiple interparticle scattering, and efficient light-harvesting. Results indicate that this as-fabricated double-layer TiO2 sphere/TNTA front-illumination dye-sensitized solar cell, which is fabricated from the TiO2 nanotube arrays with a 17.4 μm length after TiCl4 treatment, exhibits a pronounced power conversion efficiency of 7.24% under an AM1.5 G irradiation, which can be attributed to the increased incident photon-to-current conversion and light-harvesting efficiency. PMID:24304127

  20. MHC class II restricted innate-like double negative T cells contribute to optimal primary and secondary immunity to Leishmania major.

    PubMed

    Mou, Zhirong; Liu, Dong; Okwor, Ifeoma; Jia, Ping; Orihara, Kanami; Uzonna, Jude Ezeh

    2014-09-01

    Although it is generally believed that CD4(+) T cells play important roles in anti-Leishmania immunity, some studies suggest that they may be dispensable, and that MHC II-restricted CD3(+)CD4(-)CD8(-) (double negative, DN) T cells may be more important in regulating primary anti-Leishmania immunity. In addition, while there are reports of increased numbers of DN T cells in Leishmania-infected patients, dogs and mice, concrete evidence implicating these cells in secondary anti-Leishmania immunity has not yet been documented. Here, we report that DN T cells extensively proliferate and produce effector cytokines (IFN-γ, TNF and IL-17) and granzyme B (GrzB) in the draining lymph nodes and spleens of mice following primary and secondary L. major infections. DN T cells from healed mice display functional characteristics of protective anti-Leishmania memory-like cells: rapid and extensive proliferation and effector cytokines production following L. major challenge in vitro and in vivo. DN T cells express predominantly (> 95%) alpha-beta T cell receptor (αβ TCR), are Leishmania-specific, restricted mostly by MHC class II molecules and display transcriptional profile of innate-like genes. Using in vivo depletion and adoptive transfer studies, we show that DN T cells contribute to optimal primary and secondary anti-Leishmania immunity in mice. These results directly identify DN T cells as important players in effective and protective primary and secondary anti-L. major immunity in experimental cutaneous leishmaniasis. PMID:25233487

  1. Use of a small palindrome genetic marker to investigate mechanisms of double-strand-break repair in mammalian cells.

    PubMed Central

    Li, J; Baker, M D

    2000-01-01

    We examined mechanisms of mammalian homologous recombination using a gene targeting assay in which the vector-borne region of homology to the chromosome bore small palindrome insertions that frequently escape mismatch repair when encompassed within heteroduplex DNA (hDNA). Our assay permitted the product(s) of each independent recombination event to be recovered for molecular analysis. The results revealed the following: (i) vector-borne double-strand break (DSB) processing usually did not yield a large double-strand gap (DSG); (ii) in 43% of the recombinants, the results were consistent with crossover at or near the DSB; and (iii) in the remaining recombinants, hDNA was an intermediate. The sectored (mixed) genotypes observed in 38% of the recombinants provided direct evidence for involvement of hDNA, while indirect evidence was obtained from the patterns of mismatch repair (MMR). Individual hDNA tracts were either long or short and asymmetric or symmetric on the one side of the DSB examined. Clonal analysis of the sectored recombinants revealed how vector-borne and chromosomal markers were linked in each strand of individual hDNA intermediates. As expected, vector-borne and chromosomal markers usually resided on opposite strands. However, in one recombinant, they were linked on the same strand. The results are discussed with particular reference to the double-strand-break repair (DSBR) model of recombination. PMID:10757769

  2. Design of an Advanced Membrane Electrode Assembly Employing a Double-Layered Cathode for a PEM Fuel Cell.

    PubMed

    Kim, GyeongHee; Eom, KwangSup; Kim, MinJoong; Yoo, Sung Jong; Jang, Jong Hyun; Kim, Hyoung-Juhn; Cho, EunAe

    2015-12-23

    The membrane electrolyte assembly (MEA) designed in this study utilizes a double-layered cathode: an inner catalyst layer prepared by a conventional decal transfer method and an outer catalyst layer directly coated on a gas diffusion layer. The double-layered structure was used to improve the interfacial contact between the catalyst layer and membrane, to increase catalyst utilization and to modify the removal of product water from the cathode. Based on a series of MEAs with double-layered cathodes with an overall Pt loading fixed at 0.4 mg cm(-2) and different ratios of inner-to-outer Pt loading, the MEA with an inner layer of 0.3 mg Pt cm(-2) and an outer layer of 0.1 mg Pt cm(-2) exhibited the best performance. This performance was better than that of the conventional single-layered electrode by 13.5% at a current density of 1.4 A cm(-2). PMID:26630367

  3. Double-stranded RNA-dependent protein kinase is required for bone calcification in MC3T3-E1 cells in vitro.

    PubMed

    Yoshida, Kaya; Okamura, Hirohiko; Amorim, Bruna Rabelo; Ozaki, Akiko; Tanaka, Hiroaki; Morimoto, Hiroyuki; Haneji, Tatsuji

    2005-11-15

    In this study, we demonstrated that double-stranded RNA-dependent protein kinase (PKR) is required for the calcification of osteoblasts via the signal transducers and activators of transcription 1alpha (STAT1alpha) signaling in vitro. A dominant-negative mutant PKR cDNA, in which the amino acid lysine at 296 was replaced with arginine and which does not have catalytic activity, was transfected into mouse osteoblastic MC3T3-E1 cells; thereby, we established cells that stably expressed the PKR mutant gene (PKR-K/R). Phosphorylation of PKR was not stimulated by polyinosic-polycytidylic acid in the mutant cells. The PKR-K/R mutant cells exhibited up-regulated cell growth and had low alkaline phosphatase (ALP) activity. The PKR-K/R mutant cells were not able to form bone nodules in vitro. In the PKR-K/R mutant cells, runt-related gene 2 (Runx2)-mediated transcription decreased compared with the levels in the control cells. The expression of STAT1alpha protein increased and the protein was translocated to the nucleus in the PKR-K/R mutant cells. When the expression of STAT1alpha protein in PKR mutant cells was suppressed using RNAi, the activity of Runx2-mediated transcription recovered to the control level. Our results indicate that PKR is a stimulator of Runx2 transcription and is a negative modulator of STAT1alpha expression. Our findings also suggest that PKR plays important roles in the differentiation and calcification of osteoblasts by modulating STAT1alpha and/or Runx2 expression. PMID:16216244

  4. Gene conversion is strongly induced in human cells by double-strand breaks and is modulated by the expression of BCL-XL

    SciTech Connect

    Wiese, Claudia; Pierce, Andrew J.; Gauny, Stacey S.; Jasin, Maria; Kronenberg, Amy

    2001-09-25

    Homology-directed repair (HDR) of DNA double-strand breaks (DSBs) is a well-established mechanism that contributes to the maintenance of genomic stability in rodent cells, and it has been assumed that HDR is of similar importance in the repair of DSBs in human cells. However, in addition to promoting genomic stability, some outcomes of homologous recombination can be deleterious, suggesting that factors exist to regulate HDR. We previously demonstrated that overexpression of BCL-2 or BCL-xL enhanced the frequency of x-ray-induced mutations involving the TK1 locus, including loss of heterozygosity (LOH) events presumed to arise by mitotic recombination. The present study was designed to test whether HDR is a prominent DSB repair pathway in human cells, and to directly determine whether ectopic expression of BCL-xL affects HDR. We used the B-lymphoblastoid cell line TK6, which expresses wild-type TP53 and resembles normal lymphocytes in undergoing apoptosis following! genotoxic stress. U sing isogenic derivatives of TK6 cells (TK6-neo, TK6-bcl-xL), we find that a DSB in an integrated HDR reporter stimulates gene conversion 40-50-fold in TK6-neo cells, demonstrating that a DSB can be efficiently repaired by gene conversion in human cells. Significantly, DSB-induced gene conversion events are 3- to 4-fold more frequent in BCL-xL overexpressing cells. The results demonstrate that HDR plays an important role in maintaining genomic integrity in human cells and that ectopic expression of BCL-xL enhances HDR of DSBs. To our knowledge, this is the first study to highlight a function for BCL-xL in modulating DSB repair in human cells.

  5. Correlations between radiation-induced double strand breaks, cell division delay, and cyclin-dependent signaling in x-irradiated NIH3T3 fibroblasts

    NASA Astrophysics Data System (ADS)

    Cariveau, Mickael J.

    2005-07-01

    Molecular responses to radiation-induced DNA double strand breaks (DSB) are mediated by the phosphorylation of the histone variant H2AX which forms identifiable gamma-H2AX foci at the site of the DSB. This event is thought to be linked with the down-regulation of signaling proteins contributing to the checkpoints regulating cell cycle progression and, vis-a-vis , the induction of cell division delay. However, it is unclear whether this division delay is directly related to the number of DSB (gamma-H2AX foci) sustained by an irradiated cell and, if so, whether this number drives cells into cell cycle delay or apoptosis. For this reason, studies were conducted in the immortalized NIH/3T3 fibroblast cell in order to establish correlations between the temporal appearance of the gamma-H2AX foci (a DSB) and the expression of the cell cycle regulatory proteins, cyclin E, A, B1, and their cyclin kinase inhibitor, p21. Cell cycle kinetics and flow cytometry were used to establish radiation-induced division delay over a dose range of 1--6 Gy where a mitotic delay of 2.65 min/cGy was established. Correlations between the expression of cyclin E, A, B1, p21, and the generation of DSB were established in NIH/3T3 cells exposed to 2 or 4 Gy x-irradiation. The data suggest that the G1/S and S phase delay (cyclin E and cyclin A protein levels) are dependent on the dose of radiation while the G2/M (cyclin B1 protein levels) delay is dependent on the quantity of DSB sustained by the irradiated cell.

  6. Selective Cell Adhesion and Biosensing Applications of Bio-Active Block Copolymers Prepared by CuAAC/Thiol-ene Double Click Reactions.

    PubMed

    Oyman Eyrilmez, Gizem; Doran, Sean; Murtezi, Eljesa; Demir, Bilal; Odaci Demirkol, Dilek; Coskunol, Hakan; Timur, Suna; Yagci, Yusuf

    2015-09-01

    N-Acetyl-l-cysteine (NAC)-capped poly(methyl methacrylate)-b-polycaprolactone block copolymer (PMMA-b-PCL-NAC) was prepared using the previously described one-pot photoinduced sequential CuAAC/thiol-ene double click procedure. PMMA-b-PCL-NAC had previously shown good applicability as a matrix for cell adhesion of cells from the Vero cell line (African green monkey kidney epithelial). Here, in this work, PMMA-b-PCL-NAC served as an excellent immobilization matrix for biomolecule conjugation. Covalent binding of RGD (R: arginine, G: glycine, and D: aspartic acid) peptide sequence onto the PMMA-b-PCL-NAC-coated surface was performed via EDC chemistry. RGD-modified PMMA-b-PCL-NAC (PMMA-b-PCL-NAC-RGD) as a non-toxic cell proliferation platform was used for selective "integrin αvβ3-mediated cell adhesion and biosensing studies. Both optical and electrochemical techniques were used to monitor the adhesion differences between "integrin αvβ3" receptor positive and negative cell lines on to the designed biofunctional surfaces. PMID:25974890

  7. Transcription Factor Runx3 Is Induced by Influenza A Virus and Double-Strand RNA and Mediates Airway Epithelial Cell Apoptosis

    PubMed Central

    Gan, Huachen; Hao, Qin; Idell, Steven; Tang, Hua

    2015-01-01

    Influenza A virus (IAV) targets airway epithelial cells and exploits the host cell machinery to replicate, causing respiratory illness in annual epidemics and pandemics of variable severity. The high rate of antigenic drift (viral mutation) and the putative antigenic shift (reassortant strains) have raised the need to find the host cell inducible factors modulating IAV replication and its pathogenesis to develop more effective antiviral treatment. In this study, we found for the first time that transcription factor Runx3, a developmental regulator and tumor suppressor, was induced by IAV H1N1 and H3N2, viral RNA, a synthetic analog of viral double-stranded RNA (dsRNA) polyinosinic-polycytidylic acid, and type-II interferon-γ (IFNγ) in human airway epithelial cells. Whereas Runx3 was essentially not induced by type-I IFNα and type-III IFNλ, we show that Runx3 induction by IAV infection and viral RNA is mediated through the innate immune receptor MDA5 and the IκB kinase-β−NF-κB pathway. Moreover, we provide substantial evidence indicating that Runx3 plays a crucial role in airway epithelial cell apoptosis induced by IAV infection and dsRNA through the activation of extrinsic and intrinsic apoptosis pathways. Thus, we have identified Runx3 as an inducible and important transcription factor modulating IAV-induced host epithelial cell apoptosis. PMID:26643317

  8. The over-expression of the β2 catalytic subunit of the proteasome decreases homologous recombination and impairs DNA double-strand break repair in human cells.

    PubMed

    Collavoli, Anita; Comelli, Laura; Cervelli, Tiziana; Galli, Alvaro

    2011-01-01

    By a human cDNA library screening, we have previously identified two sequences coding two different catalytic subunits of the proteasome which increase homologous recombination (HR) when overexpressed in the yeast Saccharomyces cerevisiae. Here, we investigated the effect of proteasome on spontaneous HR and DNA repair in human cells. To determine if the proteasome has a role in the occurrence of spontaneous HR in human cells, we overexpressed the β2 subunit of the proteasome in HeLa cells and determined the effect on intrachromosomal HR. Results showed that the overexpression of β2 subunit decreased HR in human cells without altering the cell proteasome activity and the Rad51p level. Moreover, exposure to MG132 that inhibits the proteasome activity reduced HR in human cells. We also found that the expression of the β2 subunit increases the sensitivity to the camptothecin that induces DNA double-strand break (DSB). This suggests that the β2 subunit has an active role in HR and DSB repair but does not alter the intracellular level of the Rad51p. PMID:21660142

  9. The Over-expression of the β2 Catalytic Subunit of the Proteasome Decreases Homologous Recombination and Impairs DNA Double-Strand Break Repair in Human Cells

    PubMed Central

    Collavoli, Anita; Comelli, Laura; Cervelli, Tiziana; Galli, Alvaro

    2011-01-01

    By a human cDNA library screening, we have previously identified two sequences coding two different catalytic subunits of the proteasome which increase homologous recombination (HR) when overexpressed in the yeast Saccharomyces cerevisiae. Here, we investigated the effect of proteasome on spontaneous HR and DNA repair in human cells. To determine if the proteasome has a role in the occurrence of spontaneous HR in human cells, we overexpressed the β2 subunit of the proteasome in HeLa cells and determined the effect on intrachromosomal HR. Results showed that the overexpression of β2 subunit decreased HR in human cells without altering the cell proteasome activity and the Rad51p level. Moreover, exposure to MG132 that inhibits the proteasome activity reduced HR in human cells. We also found that the expression of the β2 subunit increases the sensitivity to the camptothecin that induces DNA double-strand break (DSB). This suggests that the β2 subunit has an active role in HR and DSB repair but does not alter the intracellular level of the Rad51p. PMID:21660142

  10. DNA double strand break repair defect and sensitivity to poly ADP-ribose polymerase (PARP) inhibition in human papillomavirus 16-positive head and neck squamous cell carcinoma

    PubMed Central

    Weaver, Alice N.; Cooper, Tiffiny S.; Rodriguez, Marcela; Trummell, Hoa Q.; Bonner, James A.; Rosenthal, Eben L.; Yang, Eddy S.

    2015-01-01

    Patients with human papillomavirus-positive (HPV+) head and neck squamous cell carcinomas (HNSCCs) have increased response to radio- and chemotherapy and improved overall survival, possibly due to an impaired DNA damage response. Here, we investigated the correlation between HPV status and repair of DNA damage in HNSCC cell lines. We also assessed in vitro and in vivo sensitivity to the PARP inhibitor veliparib (ABT-888) in HNSCC cell lines and an HPV+ patient xenograft. Repair of DNA double strand breaks (DSBs) was significantly delayed in HPV+ compared to HPV− HNSCCs, resulting in persistence of γH2AX foci. Although DNA repair activators 53BP1 and BRCA1 were functional in all HNSCCs, HPV+ cells showed downstream defects in both non-homologous end joining and homologous recombination repair. Specifically, HPV+ cells were deficient in protein recruitment and protein expression of DNA-Pk and BRCA2, key factors for non-homologous end joining and homologous recombination respectively. Importantly, the apparent DNA repair defect in HPV+ HNSCCs was associated with increased sensitivity to the PARP inhibitor veliparib, resulting in decreased cell survival in vitro and a 10–14 day tumor growth delay in vivo. These results support the testing of PARP inhibition in combination with DNA damaging agents as a novel therapeutic strategy for HPV+ HNSCC. PMID:26336991

  11. Effect of Chromatin Structure on the Extent and Distribution of DNA Double Strand Breaks Produced by Ionizing Radiation; Comparative Study of hESC and Differentiated Cells Lines

    PubMed Central

    Venkatesh, Priyanka; Panyutin, Irina V.; Remeeva, Evgenia; Neumann, Ronald D.; Panyutin, Igor G.

    2016-01-01

    Chromatin structure affects the extent of DNA damage and repair. Thus, it has been shown that heterochromatin is more protective against DNA double strand breaks (DSB) formation by ionizing radiation (IR); and that DNA DSB repair may proceed differently in hetero- and euchromatin regions. Human embryonic stem cells (hESC) have a more open chromatin structure than differentiated cells. Here, we study the effect of chromatin structure in hESC on initial DSB formation and subsequent DSB repair. DSB were scored by comet assay; and DSB repair was assessed by repair foci formation via 53BP1 antibody staining. We found that in hESC, heterochromatin is confined to distinct regions, while in differentiated cells it is distributed more evenly within the nuclei. The same dose of ionizing radiation produced considerably more DSB in hESC than in differentiated derivatives, normal human fibroblasts; and one cancer cell line. At the same time, the number of DNA repair foci were not statistically different among these cells. We showed that in hESC, DNA repair foci localized almost exclusively outside the heterochromatin regions. We also noticed that exposure to ionizing radiation resulted in an increase in heterochromatin marker H3K9me3 in cancer HT1080 cells, and to a lesser extent in IMR90 normal fibroblasts, but not in hESCs. These results demonstrate the importance of chromatin conformation for DNA protection and DNA damage repair; and indicate the difference of these processes in hESC. PMID:26729112

  12. Live imaging of induced and controlled DNA double-strand break formation reveals extremely low repair by homologous recombination in human cells.

    PubMed

    Shahar, O D; Raghu Ram, E V S; Shimshoni, E; Hareli, S; Meshorer, E; Goldberg, M

    2012-07-26

    DNA double-strand breaks (DSBs), the most hazardous DNA lesions, may result in genomic instability, a hallmark of cancer cells. The main DSB repair pathways are non-homologous end joining (NHEJ) and homologous recombination (HR). In mammalian cells, NHEJ, which can lead to inaccurate repair, predominates. HR repair (HRR) is considered accurate and is restricted to S, G2 and M phases of the cell cycle. Despite its importance, many aspects regarding HRR remain unknown. Here, we developed a novel inducible on/off switch cell system that enables, for the first time, to induce a DSB in a rapid and reversible manner in human cells. By limiting the duration of DSB induction, we found that non-persistent endonuclease-induced DSBs are rarely repaired by HR, whereas persistent DSBs result in the published HRR frequencies (non-significant HR frequency versus frequency of ∼10%, respectively). We demonstrate that these DSBs are repaired by an accurate repair mechanism, which is distinguished from HRR (most likely, error-free NHEJ). Notably, our data reveal that HRR frequencies of endonuclease-induced DSBs in human cells are >10-fold lower than what was previously estimated by prevailing methods, which resulted in recurrent DSB formation. Our findings suggest a role for HRR mainly in repairing challenging DSBs, in contrast to uncomplicated lesions that are frequently repaired by NHEJ. Preventing HR from repairing DSBs in the complex and repetitive human genome probably has an essential role in maintaining genomic stability. PMID:22105360

  13. Dual Targeting of Akt and mTORC1 Impairs Repair of DNA Double-Strand Breaks and Increases Radiation Sensitivity of Human Tumor Cells

    PubMed Central

    Holler, Marina; Grottke, Astrid; Mueck, Katharina; Manes, Julia; Jücker, Manfred

    2016-01-01

    Inhibition of mammalian target of rapamycin-complex 1 (mTORC1) induces activation of Akt. Because Akt activity mediates the repair of ionizing radiation-induced DNA double-strand breaks (DNA-DSBs) and consequently the radioresistance of solid tumors, we investigated whether dual targeting of mTORC1 and Akt impairs DNA-DSB repair and induces radiosensitization. Combining mTORC1 inhibitor rapamycin with ionizing radiation in human non-small cell lung cancer (NSCLC) cells (H661, H460, SK-MES-1, HTB-182, A549) and in the breast cancer cell line MDA-MB-231 resulted in radiosensitization of H661 and H460 cells (responders), whereas only a very slight effect was observed in A549 cells, and no effect was observed in SK-MES-1, HTB-182 or MDA-MB-231 cells (non-responders). In responder cells, rapamycin treatment did not activate Akt1 phosphorylation, whereas in non-responders, rapamycin mediated PI3K-dependent Akt activity. Molecular targeting of Akt by Akt inhibitor MK2206 or knockdown of Akt1 led to a rapamycin-induced radiosensitization of non-responder cells. Compared to the single targeting of Akt, the dual targeting of mTORC1 and Akt1 markedly enhanced the frequency of residual DNA-DSBs by inhibiting the non-homologous end joining repair pathway and increased radiation sensitivity. Together, lack of radiosensitization induced by rapamycin was associated with rapamycin-mediated Akt1 activation. Thus, dual targeting of mTORC1 and Akt1 inhibits repair of DNA-DSB leading to radiosensitization of solid tumor cells. PMID:27137757

  14. Highly Unstable Double-Diffusive Finger Convection in a Hele-Shaw Cell: Baseline Experimental Data for Evaluation of Numerical Models

    SciTech Connect

    PRINGLE,SCOTT E.; COOPER,CLAY A.; GLASS JR.,ROBERT J.

    2000-12-21

    An experimental investigation was conducted to study double-diffusive finger convection in a Hele-Shaw cell by layering a sucrose solution over a more-dense sodium chloride (NaCl) solution. The solutal Rayleigh numbers were on the order of 60,000, based upon the height of the cell (25 cm), and the buoyancy ratio was 1.2. A full-field light transmission technique was used to measure a dye tracer dissolved in the NaCl solution. They analyze the concentration fields to yield the temporal evolution of length scales associated with the vertical and horizontal finger structure as well as the mass flux. These measures show a rapid progression through two early stages to a mature stage and finally a rundown period where mass flux decays rapidly. The data are useful for the development and evaluation of numerical simulators designed to model diffusion and convection of multiple components in porous media. The results are useful for correct formulation at both the process scale (the scale of the experiment) and effective scale (where the lab-scale processes are averaged-up to produce averaged parameters). A fundamental understanding of the fine-scale dynamics of double-diffusive finger convection is necessary in order to successfully parameterize large-scale systems.

  15. Efficient Rejoining of DNA Double-Strand Breaks despite Increased Cell-Killing Effectiveness following Spread-Out Bragg Peak Carbon-Ion Irradiation.

    PubMed

    Averbeck, Nicole B; Topsch, Jana; Scholz, Michael; Kraft-Weyrather, Wilma; Durante, Marco; Taucher-Scholz, Gisela

    2016-01-01

    Radiotherapy of solid tumors with charged particles holds several advantages in comparison to photon therapy; among them conformal dose distribution in the tumor, improved sparing of tumor-surrounding healthy tissue, and an increased relative biological effectiveness (RBE) in the tumor target volume in the case of ions heavier than protons. A crucial factor of the biological effects is DNA damage, of which DNA double-strand breaks (DSBs) are the most deleterious. The reparability of these lesions determines the cell survival after irradiation and thus the RBE. Interestingly, using phosphorylated H2AX as a DSB marker, our data in human fibroblasts revealed that after therapy-relevant spread-out Bragg peak irradiation with carbon ions DSBs are very efficiently rejoined, despite an increased RBE for cell survival. This suggests that misrepair plays an important role in the increased RBE of heavy-ion radiation. Possible sources of erroneous repair will be discussed. PMID:26904506

  16. Modeling of a Hybrid System for a Lightweight Electric Vehicle with Passive-type Polymer Electrolyte Fuel Cells and Electric Double-layer Capacitors

    NASA Astrophysics Data System (ADS)

    Imanishi, Hiroyuki; Yoshii, Taichi; Nakamura, Takuji; Takada, Yogo; Wakisaka, Tomoyuki

    A simple series hybrid power system composed of passive-type polymer electrolyte fuel cells (PEFCs) and electric double-layer capacitors was adapted to a lightweight electric vehicle. In order to numerically simulate the behavior of the hybrid system, a fuel cell equivalent circuit model was applied and the model parameters were determined using an electrochemical theory and experimental results. Including this PEFC equivalent circuit model, a simulation model of the power train system (PEFCs, capacitors, motor, power controller, inertia, etc) of a lightweight electric vehicle was composed. It has been confirmed that this simulation model can represent reasonably well the dynamic behavior and energy transmission of the system in the experiment on a fixed apparatus constructed as a model of the vehicle.

  17. Measurement of DNA Double-Strand Break Yield in Human Cancer Cells by High-Current, Short-Duration Bunches of Laser-Accelerated Protons

    NASA Astrophysics Data System (ADS)

    Yogo, Akifumi; Sato, Katsutoshi; Nishikino, Masaharu; Maeda, Takuya; Sakaki, Hironao; Hori, Toshihiko; Ogura, Koichi; Nishiuchi, Mamiko; Teshima, Teruki; Nishimura, Hiroaki; Kondo, Kiminori; Bolton, Paul R.; Kawanishi, Shunichi

    2011-10-01

    To investigate the radiobiological effects of high dose rates that are attributed to high current, short bunch beam generation with laser-dreven ion acceleration, we have developed an experimental setup that uses laser-accelerated protons. In-vitro human lung cancer cells: A549 pulmonary adenocarcinoma are irradiated with a laser-accelerated proton bunches with a duration of 2×10-8 s and flux of ˜1015 cm-2 s-1, amounting to single bunch absorbed dose at the 1 Gy level. The double-strand break (DSB) yield in cell DNA is analyzed for the laser-accelerated proton beam at an average LET of 41 keV/µm.

  18. Spatial and Temporal Organisation of Multiprotein Systems of Cell Regulation and Signalling: What Can We Learn from NHEJ System of Double-Strand Break Repair?

    NASA Astrophysics Data System (ADS)

    Wu, Qian; Sibanda, Lynn; Ochi, Takashi; Bolanos-Garcia, Victor M.; Blundell, Tom L.; Chirgadze, Dimitri Y.

    Multiprotein assemblies play major roles in most pathways involved in cell regulation and signaling. Weak binary interactions are transformed co-operatively into very specific systems, which achieve sensitivity, specificity and temporal control. Due to the complexity and transience of these regulatory and signaling systems, a combination of in vivo, cell, biochemical, biophysical, and structural approaches is needed to investigate their structures and dynamics. Here we describe the architecture and spatial organisation of the complexes mediating Non-Homologous End Joining (NHEJ), one of the two major pathways involved in DNA double-strand break repair. Our example illustrates the experimental challenges and conceptual questions that are raised by studying such complex systems. We discuss the potential of using knowledge of the spatial and temporal organization of multiprotein systems not only to give insights into the mechanisms of pathway regulation but also to help in the design of chemical tools and ultimately new therapeutic agents.

  19. The mechanical properties and cytotoxicity of cell-laden double-network hydrogels based on photocrosslinkable gelatin and gellan gum biomacromolecules.

    PubMed

    Shin, Hyeongho; Olsen, Bradley D; Khademhosseini, Ali

    2012-04-01

    A major goal in the application of hydrogels for tissue engineering scaffolds, especially for load-bearing tissues such as cartilage, is to develop hydrogels with high mechanical strength. In this study, a double-network (DN) strategy was used to engineer strong hydrogels that can encapsulate cells. We improved upon previously studied double-network (DN) hydrogels by using a processing condition compatible with cell survival. The DN hydrogels were created by a two-step photocrosslinking using gellan gum methacrylate (GGMA) for the rigid and brittle first network, and gelatin methacrylamide (GelMA) for the soft and ductile second network. We controlled the degree of methacrylation of each polymer so that they obtain relevant mechanical properties as each network. The DN was formed by photocrosslinking the GGMA, diffusing GelMA into the first network, and photocrosslinking the GelMA to form the second network. The formation of the DN was examined by diffusion tests of the large GelMA molecules into the GGMA network, the resulting enhancement in the mechanical properties, and the difference in mechanical properties between GGMA/GelMA single networks (SN) and DNs. The resulting DN hydrogels exhibited the compressive failure stress of up to 6.9 MPa, which approaches the strength of cartilage. It was found that there is an optimal range of the crosslink density of the second network for high strength of DN hydrogels. DN hydrogels with a higher mass ratio of GelMA to GGMA exhibited higher strength, which shows promise in developing even stronger DN hydrogels in the future. Three dimensional (3D) encapsulation of NIH-3T3 fibroblasts and the following viability test showed the cell-compatibility of the DN formation process. Given the high strength and the ability to encapsulate cells, the DN hydrogels made from photocrosslinkable macromolecules could be useful for the regeneration of load-bearing tissues. PMID:22265786

  20. The mechanical properties and cytotoxicity of cell-laden double-network hydrogels based on photocrosslinkable gelatin and gellan gum biomacromolecules

    PubMed Central

    Shin, Hyeongho; Olsen, Bradley D.; Khademhosseini, Ali

    2012-01-01

    A major goal in the application of hydrogels for tissue engineering scaffolds, especially for load-bearing tissues such as cartilage, is to develop hydrogels with high mechanical strength. In this study, a double-network (DN) strategy was used to engineer strong hydrogels that can encapsulate cells. We improved upon previously studied double-network (DN) hydrogels by using a processing condition compatible with cell survival. The DN hydrogels were created by a two-step photocrosslinking using gellan gum methacrylate (GGMA) for the rigid and brittle first network, and gelatin methacrylamide (GelMA) for the soft and ductile second network. We controlled the degree of methacrylation of each polymer so that they obtain relevant mechanical properties as each network. The DN was formed by photocrosslinking the GGMA, diffusing GelMA into the first network, and photocrosslinking the GelMA to form the second network. The formation of the DN was examined by diffusion tests of the large GelMA molecules into the GGMA network, the resulting enhancement in the mechanical properties, and the difference in mechanical properties between GGMA/GelMA single networks (SN) and DNs. The resulting DN hydrogels exhibited the compressive failure stress of up to 6.9 MPa, which approaches the strength of cartilage. It was found that there is an optimal range of the crosslink density of the second network for high strength of DN hydrogels. DN hydrogels with a higher mass ratio of GelMA to GGMA exhibited higher strength, which shows promise in developing even stronger DN hydrogels in the future. Three dimensional (3D) encapsulation of NIH-3T3 fibroblasts and the following viability test showed the cell-compatibility of the DN formation process. Given the high strength and the ability to encapsulate cells, the DN hydrogels made from photocrosslinkable macromolecules could be useful for the regeneration of load-bearing tissues. PMID:22265786

  1. Androgen receptor in Sertoli cells regulates DNA double-strand break repair and chromosomal synapsis of spermatocytes partially through intercellular EGF-EGFR signaling

    PubMed Central

    Chen, Su-Ren; Hao, Xiao-Xia; Zhang, Yan; Deng, Shou-Long; Wang, Zhi-Peng; Wang, Yu-Qian; Wang, Xiu-Xia; Liu, Yi-Xun

    2016-01-01

    Spermatogenesis does not progress beyond the pachytene stages of meiosis in Sertoli cell-specific AR knockout (SCARKO) mice. However, further evidence of meiotic arrest and underlying paracrine signals in SCARKO testes is still lacking. We utilized co-immunostaining of meiotic surface spreads to examine the key events during meiotic prophase I. SCARKO spermatocytes exhibited a failure in chromosomal synapsis observed by SCP1/SCP3 double-staining and CREST foci quantification. In addition, DNA double-strand breaks (DSBs) were formed but were not repaired in the mutant spermatocytes, as revealed by γ-H2AX staining and DNA-dependent protein kinase (DNA-PK) activity examination. The later stages of DSB repair, such as the accumulation of the RAD51 strand exchange protein and the localization of mismatch repair protein MLH1, were correspondingly altered in SCARKO spermatocytes. Notably, the expression of factors that guide RAD51 loading onto sites of DSBs, including TEX15, BRCA1/2 and PALB2, was severely impaired when either AR was down-regulated or EGF was up-regulated. We observed that some ligands in the epidermal growth factor (EGF) family were over-expressed in SCARKO Sertoli cells and that some receptors in the EGF receptor (EGFR) family were ectopically activated in the mutant spermatocytes. When EGF-EGFR signaling was repressed to approximately normal by the specific inhibitor AG1478 in the cultured SCARKO testis tissues, the arrested meiosis was partially rescued, and functional haploid cells were generated. Based on these data, we propose that AR in Sertoli cells regulates DSB repair and chromosomal synapsis of spermatocytes partially through proper intercellular EGF-EGFR signaling. PMID:26959739

  2. Rearrangements and deletions of immunoglobulin heavy chain genes in the double-producing B cell lymphoma I.29.

    PubMed Central

    Stavnezer, J; Marcu, K B; Sirlin, S; Alhadeff, B; Hammerling, U

    1982-01-01

    The B cell lymphoma I.29 consists of a mixture of cells expressing membrane-bound immunoglobulin M (IgM) (lambda) and IgA (lambda) of identical idiotypes. Whereas most of the cells express either IgM or IgA alone, 1 to 5% of the cells in this tumor express IgM and IgA simultaneously within the cytoplasm and on the cell membrane (R. Sitia et al., J. Immunol. 127:1388-1394, 1981; R. Sitia, unpublished data). When IgM+ cells are purified from the lymphoma and passaged in mice or cultured, a portion of the cells convert to IgA+. These properties suggest that some cells of the I.29 lymphoma may undergo immunoglobulin heavy chain switching, although it is also possible that the mixed population was derived by a prior switching event in a clone of cells. We performed Southern blotting experiments on genomic DNAs isolated from populations of I.29 cells containing variable proportions of IgM+ and IgA+ cells and on a number of cell lines derived from the lymphoma. The results were consistent with the deletion model for heavy chain switching, as the IgM+ cells contained rearranged mu genes and alpha genes in the germ line configuration on both the expressed and nonexpressed heavy chain chromosomes, whereas the IgA+ cells had deleted both mu genes and contained one rearranged and one germ line alpha gene. In addition, segments of DNA located within the intervening sequence 5' to the mu gene, near the site of switch recombination, were deleted from both the expressed and the nonexpressed chromosomes. Although mu genes were deleted from both chromosomes in the IgA+ cells, the sites of DNA recombination differed on the two chromosomes. On the expressed chromosome, Smu sequences were recombined with S alpha sequences, whereas on the nonexpressed chromosome, Smu sequences were recombined with S gamma 3 sequences. Images PMID:6290869

  3. Double Layers in Astrophysics

    NASA Technical Reports Server (NTRS)

    Williams, Alton C. (Editor); Moorehead, Tauna W. (Editor)

    1987-01-01

    Topics addressed include: laboratory double layers; ion-acoustic double layers; pumping potential wells; ion phase-space vortices; weak double layers; electric fields and double layers in plasmas; auroral double layers; double layer formation in a plasma; beamed emission from gamma-ray burst source; double layers and extragalactic jets; and electric potential between plasma sheet clouds.

  4. Ion-kinetic simulations of D-3He gas-filled inertial confinement fusion target implosions with moderate to large Knudsen number

    NASA Astrophysics Data System (ADS)

    Larroche, O.; Rinderknecht, H. G.; Rosenberg, M. J.; Hoffman, N. M.; Atzeni, S.; Petrasso, R. D.; Amendt, P. A.; Séguin, F. H.

    2016-01-01

    Experiments designed to investigate the transition to non-collisional behavior in D3He-gas inertial confinement fusion target implosions display increasingly large discrepancies with respect to simulations by standard hydrodynamics codes as the expected ion mean-free-paths λc increase with respect to the target radius R (i.e., when the Knudsen number NK=λc/R grows). To take properly into account large NK's, multi-ion-species Vlasov-Fokker-Planck computations of the inner gas in the capsules have been performed, for two different values of NK, one moderate and one large. The results, including nuclear yield, reactivity-weighted ion temperatures, nuclear emissivities, and surface brightness, have been compared with the experimental data and with the results of hydrodynamical simulations, some of which include an ad hoc modeling of kinetic effects. The experimental results are quite accurately rendered by the kinetic calculations in the smaller-NK case, much better than by the hydrodynamical calculations. The kinetic effects at play in this case are thus correctly understood. However, in the higher-NK case, the agreement is much worse. The remaining discrepancies are shown to arise from kinetic phenomena (e.g., inter-species diffusion) occurring at the gas-pusher interface, which should be investigated in the future work.

  5. Ion-kinetic simulations of D-3He gas-filled inertial confinement fusion target implosions with moderate to large Knudsen number

    DOE PAGESBeta

    Larroche, O.; Rinderknecht, H. G.; Rosenberg, M. J.; Hoffman, N. M.; Atzeni, S.; Petrasso, R. D.; Amendt, P. A.; Seguin, F. H.

    2016-01-06

    Experiments designed to investigate the transition to non-collisional behavior in D3He-gas inertial confinement fusion target implosions display increasingly large discrepancies with respect to simulations by standard hydrodynamics codes as the expected ion mean-free-paths λc increase with respect to the target radius R (i.e., when the Knudsen number NK = λc/R grows). To take properly into account large NK's, multi-ion-species Vlasov-Fokker-Planck computations of the inner gas in the capsules have been performed, for two different values of NK, one moderate and one large. The results, including nuclear yield, reactivity-weighted ion temperatures, nuclear emissivities, and surface brightness, have been compared with themore » experimental data and with the results of hydrodynamical simulations, some of which include an ad hocmodeling of kinetic effects. The experimental results are quite accurately rendered by the kinetic calculations in the smaller-NK case, much better than by the hydrodynamical calculations. The kinetic effects at play in this case are thus correctly understood. However, in the higher-NK case, the agreement is much worse. Furthermore, the remaining discrepancies are shown to arise from kinetic phenomena (e.g., inter-species diffusion) occurring at the gas-pusher interface, which should be investigated in the future work.« less

  6. I-SceI-Mediated Double-Strand Break Does Not Increase the Frequency of Homologous Recombination at the Dct Locus in Mouse Embryonic Stem Cells

    PubMed Central

    Fenina, Myriam; Simon-Chazottes, Dominique; Vandormael-Pournin, Sandrine; Soueid, Jihane; Langa, Francina; Cohen-Tannoudji, Michel; Bernard, Bruno A.; Panthier, Jean-Jacques

    2012-01-01

    Targeted induction of double-strand breaks (DSBs) at natural endogenous loci was shown to increase the rate of gene replacement by homologous recombination in mouse embryonic stem cells. The gene encoding dopachrome tautomerase (Dct) is specifically expressed in melanocytes and their precursors. To construct a genetic tool allowing the replacement of Dct gene by any gene of interest, we generated an embryonic stem cell line carrying the recognition site for the yeast I-SceI meganuclease embedded in the Dct genomic segment. The embryonic stem cell line was electroporated with an I-SceI expression plasmid, and a template for the DSB-repair process that carried sequence homologies to the Dct target. The I-SceI meganuclease was indeed able to introduce a DSB at the Dct locus in live embryonic stem cells. However, the level of gene targeting was not improved by the DSB induction, indicating a limited capacity of I-SceI to mediate homologous recombination at the Dct locus. These data suggest that homologous recombination by meganuclease-induced DSB may be locus dependent in mammalian cells. PMID:22761925

  7. Inhibition of NF-κB and DNA double-strand break repair by DMAPT sensitizes non-small-cell lung cancers to X-rays.

    PubMed

    Estabrook, Neil C; Chin-Sinex, Helen; Borgmann, Anthony J; Dhaemers, Ryan M; Shapiro, Ronald H; Gilley, David; Huda, Nazmul; Crooks, Peter; Sweeney, Christopher; Mendonca, Marc S

    2011-12-15

    We investigated the efficacy and mechanism of dimethylaminoparthenolide (DMAPT), an NF-κB inhibitor, to sensitize human lung cancer cells to X-ray killing in vitro and in vivo. We tested whether DMAPT increased the effectiveness of single and fractionated X-ray treatment through inhibition of NF-κB and/or DNA double-strand break (DSB) repair. Treatment with DMAPT decreased plating efficiency, inhibited constitutive and radiation-induced NF-κB binding activity, and enhanced radiation-induced cell killing by dose modification factors of 1.8 and 1.4 in vitro. X-ray fractionation demonstrated that DMAPT inhibited split-dose recovery/repair, and neutral DNA comet assays confirmed that DMAPT altered the fast and slow components of X-ray-induced DNA DSB repair. Knockdown of the NF-κB family member p65 by siRNA increased radiation sensitivity and completely inhibited split-dose recovery in a manner very similar to DMAPT treatment. The data suggest a link between inhibition of NF-κB and inhibition of DSB repair by DMAPT that leads to enhancement of X-ray-induced cell killing in vitro in non-small-cell lung cancer cells. Studies of A549 tumor xenografts in nude mice demonstrated that DMAPT enhanced X-ray-induced tumor growth delay in vivo. PMID:22019440

  8. Isolation of mammalian cell mutants that are X-ray sensitive, impaired in DNA double-strand break repair and defective for V(D)J recombination.

    PubMed

    Lee, S E; Pulaski, C R; He, D M; Benjamin, D M; Voss, M; Um, J; Hendrickson, E A

    1995-05-01

    The Chinese hamster lung V79-4 cell line was infected with a Moloney murine leukemia retrovirus and the infected cells were subsequently screened for mutants that were sensitive to X-rays using a toothpicking/96-well replica plating technique. Four independent mutants that were sensitive to X-irradiation (sxi-1 to sxi-4) were isolated from 9000 retrovirally infected colonies. A pulse-field gel electrophoresis (PFGE) assay demonstrated that all of the sxi mutants were impaired in DNA double-strand break (DSB) repair, thus providing a molecular explanation for the observed X-ray sensitivity. Interestingly, additional PFGE experiments demonstrated that for any given X-ray dose all of the mutants incurred more DNA DSBs than the parental V79-4 cell line indicating there may be some inherent fragility to sxi chromosomes. Cross-sensitivity to other DNA-damaging agents including bleomycin, mitomycin C and methyl methanesulfonate indicated that sxi-2, sxi-3 and sxi-4 appear to be specifically hypersensitive to genotoxic agents that cause DNA DSBs, whereas sxi-1 appeared to be hypersensitive to multiple types of DNA lesions. Lastly, in preliminary experiments all of the sxi mutants demonstrated an inability to carry out V(D)J recombination, a somatic DNA rearrangement process required for the assembly of lymphoid antigen receptor genes. Thus, the sxi cell lines have interesting phenotypes which should make them valuable tools for unraveling the mechanism(s) of DNA DSB repair and recombination in mammalian cells. PMID:7537861

  9. DNA Ligase IV and Artemis Act Cooperatively to Suppress Homologous Recombination in Human Cells: Implications for DNA Double-Strand Break Repair

    PubMed Central

    Kurosawa, Aya; Saito, Shinta; So, Sairei; Hashimoto, Mitsumasa; Iwabuchi, Kuniyoshi; Watabe, Haruka; Adachi, Noritaka

    2013-01-01

    Nonhomologous end-joining (NHEJ) and homologous recombination (HR) are two major pathways for repairing DNA double-strand breaks (DSBs); however, their respective roles in human somatic cells remain to be elucidated. Here we show using a series of human gene-knockout cell lines that NHEJ repairs nearly all of the topoisomerase II- and low-dose radiation-induced DNA damage, while it negatively affects survival of cells harbouring replication-associated DSBs. Intriguingly, we find that loss of DNA ligase IV, a critical NHEJ ligase, and Artemis, an NHEJ factor with endonuclease activity, independently contribute to increased resistance to replication-associated DSBs. We also show that loss of Artemis alleviates hypersensitivity of DNA ligase IV-null cells to low-dose radiation- and topoisomerase II-induced DSBs. Finally, we demonstrate that Artemis-null human cells display increased gene-targeting efficiencies, particularly in the absence of DNA ligase IV. Collectively, these data suggest that DNA ligase IV and Artemis act cooperatively to promote NHEJ, thereby suppressing HR. Our results point to the possibility that HR can only operate on accidental DSBs when NHEJ is missing or abortive, and Artemis may be involved in pathway switching from incomplete NHEJ to HR. PMID:23967291

  10. DNA double-strand-break repair in higher eukaryotes and its role in genomic instability and cancer: Cell cycle and proliferation-dependent regulation.

    PubMed

    Mladenov, Emil; Magin, Simon; Soni, Aashish; Iliakis, George

    2016-06-01

    Eukaryotic cells respond to DNA damage by activating a comprehensive network of biochemical pathways that enable damage recognition and initiate responses leading to repair, apoptosis/autophagy or senescence. This network of responses is commonly described as the "DNA damage response" (DDR). Among the plethora of lesions generated in the DNA from various physical and chemical agents in the environment and in the cell, DNA double strand breaks (DSBs) and DNA replication stress (RS) are the most severe and induce strong DDR, as they bear high risk for cell death, or genomic alterations ultimately causing cancer. Here, we focus on DSBs and provide a state-of-the-art review of the molecular underpinnings of repair pathways that process DSBs in higher eukaryotes, their strengths and limitations, as well as aspects of repair pathway choice and hierarchy. Furthermore, we discuss the regulation of DSB repair pathways throughout the cell cycle and by processes affecting the proliferative state of the cell. We review the role of growth factors and their receptors in the regulation of each DSB repair pathway and discuss aspects of systemic regulation of DNA repair. PMID:27016036

  11. Autologous olfactory mucosal cell transplants in clinical spinal cord injury: a randomized double-blinded trial in a canine translational model

    PubMed Central

    Granger, Nicolas; Blamires, Helen; Franklin, Robin J. M.

    2012-01-01

    This study was designed to determine whether an intervention proven effective in the laboratory to ameliorate the effects of experimental spinal cord injury could provide sufficient benefit to be of value to clinical cases. Intraspinal olfactory ensheathing cell transplantation improves locomotor outcome after spinal cord injury in ‘proof of principle’ experiments in rodents, suggesting the possibility of efficacy in human patients. However, laboratory animal spinal cord injury cannot accurately model the inherent heterogeneity of clinical patient cohorts, nor are all aspects of their spinal cord function readily amenable to objective evaluation. Here, we measured the effects of intraspinal transplantation of cells derived from olfactory mucosal cultures (containing a mean of ∼50% olfactory ensheathing cells) in a population of spinal cord–injured companion dogs that accurately model many of the potential obstacles involved in transition from laboratory to clinic. Dogs with severe chronic thoracolumbar spinal cord injuries (equivalent to ASIA grade ‘A’ human patients at ∼12 months after injury) were entered into a randomized double-blinded clinical trial in which they were allocated to receive either intraspinal autologous cells derived from olfactory mucosal cultures or injection of cell transport medium alone. Recipients of olfactory mucosal cell transplants gained significantly better fore–hind coordination than those dogs receiving cell transport medium alone. There were no significant differences in outcome between treatment groups in measures of long tract functionality. We conclude that intraspinal olfactory mucosal cell transplantation improves communication across the damaged region of the injured spinal cord, even in chronically injured individuals. However, we find no evidence for concomitant improvement in long tract function. PMID:23169917

  12. Significant increase in Young's modulus of ATDC5 cells during chondrogenic differentiation induced by PAMPS/PDMAAm double-network gel: comparison with induction by insulin.

    PubMed

    Maeda, Eijiro; Tsutsumi, Takehiro; Kitamura, Nobuto; Kurokawa, Takayuki; Ping Gong, Jian; Yasuda, Kazunori; Ohashi, Toshiro

    2014-10-17

    A double-network (DN) gel, which was composed of poly(2-acrylamido-2-methylpropanesulfonic acid) and poly(N,N'-dimethyl acrylamide) (PAMPS/PDMAAm), has the potential to induce chondrogenesis both in vitro and in vivo. The present study investigated the biomechanical and biological responses of chondrogenic progenitor ATDC5 cells cultured on the DN gel. ATDC5 cells were cultured on a polystyrene surface without insulin (Culture 1) and with insulin (Culture 2), and on the DN gel without insulin (Culture 3). The cultured cells were evaluated using micropipette aspiration for cell Young's modulus and qPCR for gene expression of chondrogenic and actin organization markers on days 3, 7 and 14. On day 3, the cells in Culture 3 formed nodules, in which the cells exhibited an actin cortical layer inside them, and gene expression of type-II collagen, aggrecan, and SOX9 was significantly higher in Culture 3 than Cultures 1 and 2 (p<0.05). Young's modulus in Culture 3 was significantly higher than that in Culture 1 throughout the testing period (p<0.05) and that in Culture 2 on day 14 (p<0.01). There was continuous expression of actin organization markers in Culture 3. This study highlights that the cells on the DN gel increased the modulus and mRNA expression of chondrogenic markers at an earlier time point with a greater magnitude compared to those on the polystyrene surface with insulin. This study also demonstrates a possible strong interrelation among alteration of cell mechanical properties, changes in actin organization and the induction of chondrogenic differentiation. PMID:25110167

  13. Double screening

    NASA Astrophysics Data System (ADS)

    Gratia, Pierre; Hu, Wayne; Joyce, Austin; Ribeiro, Raquel H.

    2016-06-01

    Attempts to modify gravity in the infrared typically require a screening mechanism to ensure consistency with local tests of gravity. These screening mechanisms fit into three broad classes; we investigate theories which are capable of exhibiting more than one type of screening. Specifically, we focus on a simple model which exhibits both Vainshtein and kinetic screening. We point out that due to the two characteristic length scales in the problem, the type of screening that dominates depends on the mass of the sourcing object, allowing for different phenomenology at different scales. We consider embedding this double screening phenomenology in a broader cosmological scenario and show that the simplest examples that exhibit double screening are radiatively stable.

  14. Microdissected double-minute DNA detects variable patterns of chromosomal localizations and multiple abundantly expressed transcripts in normal and leukemic cells

    SciTech Connect

    Sen, S.; Zhou, Hongyi; Stass, S.A.; Sen, P. ); Mulac-Jericevic, B.; Pirrotta, V. )

    1994-02-01

    Double-minute (dm) chromosomes are cytogenetically resolvable DNA amplification-mediating acentric extrachromosomal structures that are commonly seen in primary tumors, tumor cell lines, and drug-resistant cells grown in vitro. Selective isolation of dm DNAs with standard molecular biological techniques is difficult, and thus, detailed studies to elucidate their structure, site of chromosomal origin, and chromosomal reintegration patterns have been limited. In those instances in which a gene has been localized on dms, characterization of the remainder of the DNA, which far exceeds the size of the gene identified, has remained inconclusive. dms seen in the acute myeloid leukemia cell line HL-60 have been shown to harbor the c-myc protooncogene. In this paper, the authors report the successful isolation of the dm-specific DNAs from these cells by the microdissection/polymerase chain reaction technique and demonstrate that the dm DNAs derived from a single discrete normal chromosome segment 8q24.1-q24.2 reintegrate at various specific locations in the leukemic cells. The microdissected dm DNA detects multiple abundantly expressed transcripts distinct from c-myc mRNA on Northern blots. By devising a [open quotes]transcript selection[close quotes] strategy, they cloned the partial genomic sequence of a gene from the microdissected DNA that encodes two of these RNAs. This strategy will be generally applicable for rapid cloning of unknown amplified genes harbored on dms. With DNA from 20 microdissected dms, they constructed a genomic library of about 20,000 recombinant microclones with an average insert size of about 450 bp. The microclones should help in isolating corresponding yeast artificial chromosome clones for high-resolution physical mapping of dms in HL-60 cells. Furthermore, application of the microdissection technique appears to be an extremely feasible approach to characterization of dms in other cell types. 42 refs., 6 figs., 1 tab.

  15. Secretory glycoconjugates of a mucin-synthesizing human colonic adenocarcinoma cell line. Analysis using double labeling with lectins.

    PubMed

    Phillips, T E; Frisch, E B

    1990-01-01

    Lectins were used to characterize mucin glycoproteins and other secretory glycoconjugates synthesized by a human colon adenocarcinoma-derived cell line which expresses a goblet cell phenotype. Despite being clonally derived, HT29-18N2 (N2) cells, like normal goblet cells in situ were heterogeneous in their glycosylation of mucin. Only wheat-germ agglutinin, which recognizes N-acetylglucosamine and sialic acid residues, and succinylated wheatgerm agglutinin, which binds N-acetylglucosamine, stained the contents of all secretory granules in all N2 goblet cells. The N-acetylgalactosamine binding lectins Dolichos biflorus and Glycine max stained 20% and 21% of N2 goblet cells respectively. Ricinus communis I, a galactose-binding lectin, stained 67% of N2 goblet cells although staining by another galactose-binding lectin, Bandeiraea simplicifolia I, was limited to 19%. Peanut agglutinin, a lectin whose Gal(beta 1-3)GalNAc binding site is not present on mucins produced in the normal colon but which is found on most mucins of cancerous colonic epithelia, stained 68% of the cells. Ulex europeus I, a fucose-binding lectin, did not stain any N2 goblet cells. Four lectins (Lens culinaris, Pisum sativum, Phaseolus vulgaris E, Phaseolus vulgaris L) which recognize sugars normally present only in N-linked oligosaccharides stained up to 38% of N2 goblet cells. The binding of these lectins indicates either both O-linked and N-linked oligosaccharide chains are present on the mucin protein backbone or the co-existence of non-mucin N-linked glycoproteins and O-linked mucins within the goblet cell secretory granule. PMID:2312359

  16. Does pretreatment of bone marrow mesenchymal stem cells with 5-azacytidine or double intravenous infusion improve their therapeutic potential for dilated cardiomyopathy?

    PubMed Central

    Yang, Sirui; Piao, Jinhua; Jin, Lianhua; Zhou, Yan

    2013-01-01

    Background This study was designed to investigate whether pretreatment of bone marrow mesenchymal stem cells (BMSCs) with 5-azacytidine (5-aza) or double intravenous infusion could enhance their therapeutic potential for dilated cardiomyopathy (DCM). Material/Methods BMSCs were cultured for 2 weeks in the presence or absence of 5-aza and DCM serum. The cultured BMSCs (Groups 1 and 2), 5-aza-induced BMSCs (Groups 3 and 4), and medium alone (model control) were transplanted into 80 female Wistar rats by intravenous tail vein injection. Double infusion of BMSCs with 1-day time-interval was carried out in Groups 2 and 4. Postmortem histological analysis and evaluation of heart function were performed at 4 weeks post-transplantation. Results Some transplanted BMSCs engrafted into myocardial tissue and were positive for cardiac marker troponin T. The hearts containing transplanted BMSCs secreted a larger amount of vascular endothelial growth factor. Cardiac function parameters and serum level of brain natriuretic peptide (BNP) did not differ among Groups 1, 3, and the model control. As compared with model control, BMSC transplantation in Groups 2 and 4 significantly decreased the serum level of BNP and improved cardiac contractile function, as evidenced by reduced left ventricular end-diastolic and end-systolic diameter, elevated ejection fraction, and fractional shortening. Conclusions BMSC transplantation is a promising strategy for the treatment of DCM. Pretreatment of BMSCs with 5-aza and DCM serum does not enhance their therapeutic efficacy, and the double intravenous BMSC infusion method is superior to single infusion for preserving cardiac contractile function in a rat model of DCM. PMID:23314418

  17. In Vitro Expansion of Bone Marrow Derived Mesenchymal Stem Cells Alters DNA Double Strand Break Repair of Etoposide Induced DNA Damage

    PubMed Central

    Hare, Ian; Gencheva, Marieta; Evans, Rebecca; Fortney, James; Piktel, Debbie; Vos, Jeffrey A.; Howell, David; Gibson, Laura F.

    2016-01-01

    Mesenchymal stem cells (MSCs) are of interest for use in diverse cellular therapies. Ex vivo expansion of MSCs intended for transplantation must result in generation of cells that maintain fidelity of critical functions. Previous investigations have identified genetic and phenotypic alterations of MSCs with in vitro passage, but little is known regarding how culturing influences the ability of MSCs to repair double strand DNA breaks (DSBs), the most severe of DNA lesions. To investigate the response to DSB stress with passage in vitro, primary human MSCs were exposed to etoposide (VP16) at various passages with subsequent evaluation of cellular damage responses and DNA repair. Passage number did not affect susceptibility to VP16 or the incidence and repair kinetics of DSBs. Nonhomologous end joining (NHEJ) transcripts showed little alteration with VP16 exposure or passage; however, homologous recombination (HR) transcripts were reduced following VP16 exposure with this decrease amplified as MSCs were passaged in vitro. Functional evaluations of NHEJ and HR showed that MSCs were unable to activate NHEJ repair following VP16 stress in cells after successive passage. These results indicate that ex vivo expansion of MSCs alters their ability to perform DSB repair, a necessary function for cells intended for transplantation. PMID:26880992

  18. A laser-plasma-produced soft X-ray laser at 89 eV generates DNA double-strand breaks in human cancer cells.

    PubMed

    Sato, Katsutoshi; Nishikino, Masaharu; Kawachi, Tetsuya; Shimokawa, Takashi; Imai, Takashi; Teshima, Teruki; Nishimura, Hiroaki; Kando, Masaki

    2015-07-01

    While it has been expected that X-ray laser will be widely applied to biomedical studies, this has not been achieved to date and its biological effects such as DNA damage have not been evaluated. As a first step for its biological application, we developed a culture cell irradiation system, particularly designed for a plasma-driven soft X-ray laser pulse, to investigate whether the soft X-ray laser is able to induce DNA double strand breaks (DSBs) in living cells or not. The human adenocarcimona cell line A549 was irradiated with the soft X-ray laser at a photon energy of 89 eV and the repair focus formation of the DSBs was assessed by immunofluorescence staining with antiphosphorylated DNA-PKcs (p-DNA-PKcs), ATM (p-ATM) and γ-H2AX antibody. The p-DNA-PKcs, ATM, and γ-H2AX foci were clearly identified after soft X-ray laser irradiation. Furthermore, the increase in the X-ray laser shot number, even from a single shot, results in the increase in p-DNA-PKcs foci. These results are the first evidence that the 89 eV soft X-ray laser is able to induce DSB in living cells. Our study demonstrated that this irradiation system is a useful tool for investigating the radiobiological effect of soft X-ray laser. PMID:25862698

  19. A laser-plasma–produced soft X-ray laser at 89 eV generates DNA double-strand breaks in human cancer cells

    PubMed Central

    Sato, Katsutoshi; Nishikino, Masaharu; Kawachi, Tetsuya; Shimokawa, Takashi; Imai, Takashi; Teshima, Teruki; Nishimura, Hiroaki; Kando, Masaki

    2015-01-01

    While it has been expected that X-ray laser will be widely applied to biomedical studies, this has not been achieved to date and its biological effects such as DNA damage have not been evaluated. As a first step for its biological application, we developed a culture cell irradiation system, particularly designed for a plasma-driven soft X-ray laser pulse, to investigate whether the soft X-ray laser is able to induce DNA double strand breaks (DSBs) in living cells or not. The human adenocarcimona cell line A549 was irradiated with the soft X-ray laser at a photon energy of 89 eV and the repair focus formation of the DSBs was assessed by immunofluorescence staining with antiphosphorylated DNA-PKcs (p-DNA-PKcs), ATM (p-ATM) and γ-H2AX antibody. The p-DNA-PKcs, ATM, and γ-H2AX foci were clearly identified after soft X-ray laser irradiation. Furthermore, the increase in the X-ray laser shot number, even from a single shot, results in the increase in p-DNA-PKcs foci. These results are the first evidence that the 89 eV soft X-ray laser is able to induce DSB in living cells. Our study demonstrated that this irradiation system is a useful tool for investigating the radiobiological effect of soft X-ray laser. PMID:25862698

  20. Double Blockade of Glioma Cell Proliferation and Migration by Temozolomide Conjugated with NPPB, a Chloride Channel Blocker.

    PubMed

    Park, Miri; Song, Chiman; Yoon, Hojong; Choi, Kee-Hyun

    2016-03-16

    Glioblastoma is the most common and aggressive primary malignant brain tumor. Temozolomide (TMZ), a chemotherapeutic agent combined with radiation therapy, is used as a standard treatment. The infiltrative nature of glioblastoma, however, interrupts effective treatment with TMZ and increases the tendency to relapse. Voltage-gated chloride channels have been identified as crucial regulators of glioma cell migration and invasion by mediating cell shape and volume change. Accordingly, chloride current inhibition by 5-nitro-2-(3-phenylpropylamino)-benzoate (NPPB), a chloride channel blocker, suppresses cell movement by diminishing the osmotic cell volume regulation. In this study, we developed a novel compound, TMZ conjugated with NPPB (TMZ-NPPB), as a potential anticancer drug. TMZ-NPPB blocked chloride currents in U373MG, a severely invasive human glioma cell line, and suppressed migration and invasion of U373MG cells. Moreover, TMZ-NPPB exhibited DNA modification activity similar to that of TMZ, and surprisingly showed remarkably enhanced cytotoxicity relative to TMZ by inducing apoptotic cell death via DNA damage. These findings indicate that TMZ-NPPB has a dual function in blocking both proliferation and migration of human glioma cells, thereby suggesting its potential to overcome challenges in current glioblastoma therapy. PMID:26711895

  1. Effects of Cationic Microbubble Carrying CD/TK Double Suicide Gene and αVβ3 Integrin Antibody in Human Hepatocellular Carcinoma HepG2 Cells

    PubMed Central

    Li, Jiale; Zhou, Ping; Li, Lan; Zhang, Yan; Shao, Yang; Tang, Li; Tian, Shuangming

    2016-01-01

    Objective Hepatocellular carcinoma (HCC), mostly derived from hepatitis or cirrhosisis, is one of the most common types of liver cancer. T-cell mediated immune response elicited by CD/TK double suicide gene has shown a substantial antitumor effect in HCC. Integrin αVβ3 over expresssion has been suggested to regulate the biology behavior of HCC. In this study, we investigated the strategy of incorporating CD/TK double suicide gene and anti-αVβ3 integrin monoclonal antibodies into cationic microbubbles (CMBsαvβ3), and evaluated its killing effect in HCC cells. Methods To improve the transfection efficiency of targeted CD/TK double suicide gene, we adopted cationic microbubbles (CMBs), a cationic delivery agent with enhanced DNA-carrying capacity. The ultrasound and high speed shearing method was used to prepare the non-targeting cationic microbubbles (CMBs). Using the biotin-avidin bridge method, αVβ3 integrin antibody was conjugated to CMBs, and CMBsαvβ3 was generated to specifically target to HepG2 cells. The morphology and physicochemical properties of the CMBsαvβ3 was detected by optical microscope and zeta detector. The conjugation of plasmid and the antibody in CMBsαvβ3 were examined by immunofluorescent microscopy and flow cytometry. The binding capacities of CMBsαvβ3 and CMBs to HCC HepG2 and normal L-02 cells were compared using rosette formation assay. To detect EGFP fluorescence and examine the transfection efficiencies of CMBsαvβ3 and CMBs in HCC cells, fluorescence microscope and contrast-enhanced sonography were adopted. mRNA and protein level of CD/TK gene were detected by RT-PCR and Western blot, respectively. To evaluate the anti-tumor effect of CMBsαvβ3, HCC cells with CMBsαvβ3 were exposed to 5-flurocytosine / ganciclovir (5-FC/GCV). Then, cell cycle distribution after treatment were detected by PI staining and flow cytometry. Apoptotic cells death were detected by optical microscope and assessed by MTT assay and TUNEL

  2. Nanocrystal-based electrochemiluminescence sensor for cell detection with Au nanoparticles and isothermal circular double-assisted signal amplification.

    PubMed

    Dai, Pan-Pan; Li, Jin-Yi; Yu, Tao; Xu, Jing-Juan; Chen, Hong-Yuan

    2015-08-15

    Here we have developed a sensitive cancer cell amplified detection method which combined Au NPs enhanced electrochemiluminescence (ECL) of CdS nanocrystals (NCs) film, with isothermal circular amplification reaction of polymerase. In DNA circular amplification detection system, hairpin DNA beacon/Au NPs composite modified CdS NCs film was used as an ECL emitter. Messenger DNA is hybridized with the aptamer modified on magnetic beads (MBs) to form MB-Au bioconjugates. In the presence of HL-60 cell, the aptamer would conjugate with the glycoprotein at cell surface and messenger DNA sequence would be released. The released messenger DNA sequence was then introduced into the cycle amplification system to trigger circular polymerizations. This assay allows us to determine the released messenger DNA equivalent to 10 cells and exhibits a significant specificity for HL-60 cells. PMID:25966387

  3. High performance planar p-i-n perovskite solar cells with crown-ether functionalized fullerene and LiF as double cathode buffer layers

    NASA Astrophysics Data System (ADS)

    Liu, Xiaodong; Lei, Ming; Zhou, Yi; Song, Bo; Li, Yongfang

    2015-08-01

    Double cathode buffer layers (CBLs) composed of fullerene derivative functionalized with a crown-ether end group in its side chain (denoted as PCBC) and a LiF layer were introduced between the PCBM acceptor layer and the top cathode in planar p-i-n perovskite solar cells (pero-SCs) based on CH3NH3PbI3-XClX. The devices with the PCBC/LiF double CBLs showed significant improvements in power conversion efficiency (PCE) and long-term stability when compared to the device with LiF single CBL. Through optimizing the spin-coating speed of PCBC, a maximum PCE of 15.53% has been achieved, which is approximately 15% higher than that of the device with single LiF CBL. The remarkable improvement in PCE can be attributed to the formation of a better ohmic contact in the CBL between PCBC and LiF/Al electrode arising from the dipole moment of PCBC, leading to the enhanced fill factor and short-circuit current density (Jsc). Besides the PCE, the long-term stability of the devices with PCBC interlayer is also superior to that of the device with LiF single CBL, which is due to the more effective protection for the perovskite/PCBM interface.

  4. High performance planar p-i-n perovskite solar cells with crown-ether functionalized fullerene and LiF as double cathode buffer layers

    SciTech Connect

    Liu, Xiaodong; Zhou, Yi E-mail: songbo@suda.edu.cn Song, Bo E-mail: songbo@suda.edu.cn; Lei, Ming; Li, Yongfang E-mail: songbo@suda.edu.cn

    2015-08-10

    Double cathode buffer layers (CBLs) composed of fullerene derivative functionalized with a crown-ether end group in its side chain (denoted as PCBC) and a LiF layer were introduced between the PCBM acceptor layer and the top cathode in planar p-i-n perovskite solar cells (pero-SCs) based on CH{sub 3}NH{sub 3}PbI{sub 3−X}Cl{sub X}. The devices with the PCBC/LiF double CBLs showed significant improvements in power conversion efficiency (PCE) and long-term stability when compared to the device with LiF single CBL. Through optimizing the spin-coating speed of PCBC, a maximum PCE of 15.53% has been achieved, which is approximately 15% higher than that of the device with single LiF CBL. The remarkable improvement in PCE can be attributed to the formation of a better ohmic contact in the CBL between PCBC and LiF/Al electrode arising from the dipole moment of PCBC, leading to the enhanced fill factor and short-circuit current density (J{sub sc}). Besides the PCE, the long-term stability of the devices with PCBC interlayer is also superior to that of the device with LiF single CBL, which is due to the more effective protection for the perovskite/PCBM interface.

  5. Double targeting, controlled release and reversible delivery of daunorubicin to cancer cells by polyvalent aptamers-modified gold nanoparticles.

    PubMed

    Taghdisi, Seyed Mohammad; Danesh, Noor Mohammad; Lavaee, Parirokh; Emrani, Ahmad Sarreshtehdar; Hassanabad, Koroush Yousefi; Ramezani, Mohammad; Abnous, Khalil

    2016-04-01

    Clinical use of daunorubicin (Dau) in treatment of leukemia has been restricted because of its cardiotoxicity. Targeted delivery of anticancer drugs could decrease their off-target effects and enhance their efficacy. In this study a modified polyvalent aptamers (PA)-Daunorubicin (Dau)-Gold nanoparticles (AuNPs) complex was designed and its efficacy was assessed in Molt-4 cells (human acute lymphoblastic leukemia T-cell, target). Dau was efficiently loaded (10.5 μM) onto 1mL of PA-modified AuNPs. Dau was released from the PA-Dau-AuNPs complex in a pH-sensitive manner (faster release at pH5.5). The results of flow cytometry analysis indicated that the PA-Dau-AuNPs complex was efficiently internalized into target cells, but not into nontarget cells. The results of MTT assay were consistent with the internalization data. PA-Dau-AuNPs complex had less cytotoxicity in U266 cells compared to Dau alone and even Apt-Dau-AuNPs complex. The PA-Dau-AuNPs complex had more cytotoxicity in Molt-4 cells compared to Dau alone and even Apt-Dau-AuNPs complex. Cytotoxicity of PA-Dau-AuNPs complex was effectively antagonized using antisense of polyvalent aptamers. In conclusion, the designed drug delivery system inherited the properties of efficient drug loading, tumor targeting, pH-dependent drug release and controllable delivery of Dau to tumor cells. PMID:26838906

  6. Double-stranded microRNA mimics can induce length- and passenger strand-dependent effects in a cell type-specific manner.

    PubMed

    Goldgraben, Mae A; Russell, Roslin; Rueda, Oscar M; Caldas, Carlos; Git, Anna

    2016-02-01

    MicroRNAs are short (17-26) noncoding RNAs driving or modulating physiological and pathological cellular events. Overexpression of miR-155 is pathogenic in B-cell malignancy but was also reported in a number of solid tumors-in particular, in breast cancer, where its role remains unclear and often contradictory. Using representative cell line models, we sought to determine whether the discrepant miR-155 effects in breast cancer could be explained by the heterogeneity of the disease. The growth of six breast cancer cell lines transfected with several miRNA mimics was analyzed. We found MCF-7 cell growth to be inhibited by miR-155 and miR-145 mimics, both 23-nt long, but not by a number of shorter mimics, including a universal commercial negative control. Microarray and Western blot analyses revealed induction of apoptosis, associated with interferon-β after activation of the double-stranded RNA sensor pathway. 3' Trimming of the miRNA mimics to 21 nt substantially reduced their growth-inhibitory potency. Mutating the canonical seed of the miR-155 mimic had no effect on the induced inhibition, which was abolished by mutating the miRNA seed of the artificial passenger strand. A panel of breast cancer cell lines showed a wide range of sensitivities to 23-mer mimics, broadly consistent with the sensitivity of the cell lines to Poly (I:C). We demonstrate two sources for nonspecific in vitro effects by miRNA mimics: duplex length and the artificial passenger strand. We highlight the danger of a universal 21-mer negative control and the importance of using matched seed mutants for reliable interpretation of phenotypes. PMID:26670622

  7. Double-stranded microRNA mimics can induce length- and passenger strand–dependent effects in a cell type–specific manner

    PubMed Central

    Goldgraben, Mae A.; Russell, Roslin; Rueda, Oscar M.; Caldas, Carlos; Git, Anna

    2016-01-01

    MicroRNAs are short (17–26) noncoding RNAs driving or modulating physiological and pathological cellular events. Overexpression of miR-155 is pathogenic in B-cell malignancy but was also reported in a number of solid tumors—in particular, in breast cancer, where its role remains unclear and often contradictory. Using representative cell line models, we sought to determine whether the discrepant miR-155 effects in breast cancer could be explained by the heterogeneity of the disease. The growth of six breast cancer cell lines transfected with several miRNA mimics was analyzed. We found MCF-7 cell growth to be inhibited by miR-155 and miR-145 mimics, both 23-nt long, but not by a number of shorter mimics, including a universal commercial negative control. Microarray and Western blot analyses revealed induction of apoptosis, associated with interferon-β after activation of the double-stranded RNA sensor pathway. 3′ Trimming of the miRNA mimics to 21 nt substantially reduced their growth-inhibitory potency. Mutating the canonical seed of the miR-155 mimic had no effect on the induced inhibition, which was abolished by mutating the miRNA seed of the artificial passenger strand. A panel of breast cancer cell lines showed a wide range of sensitivities to 23-mer mimics, broadly consistent with the sensitivity of the cell lines to Poly (I:C). We demonstrate two sources for nonspecific in vitro effects by miRNA mimics: duplex length and the artificial passenger strand. We highlight the danger of a universal 21-mer negative control and the importance of using matched seed mutants for reliable interpretation of phenotypes. PMID:26670622

  8. Characterization of a double-CRD-mutated Gal-8 recombinant protein that retains co-stimulatory activity on antigen-specific T-cell response.

    PubMed

    Schroeder, Matías Nicolás; Tribulatti, María Virginia; Carabelli, Julieta; André-Leroux, Gwenaëlle; Caramelo, Julio Javier; Cattaneo, Valentina; Campetella, Oscar

    2016-04-01

    Galectins (Gals) constitute a family of mammalian lectins with affinity for β-galactosides, characterized by the presence of conserved CRDs (carbohydrate-recognition domains). We have found previously that Gal-8, from the tandem-repeat group with two linked CRDs, exerts two separate actions on CD4(+)T-cells: antigen-independent proliferation and, at lower concentration, antigen-specific co-stimulation. Whereas proliferation can be ascribed to the pro-inflammatory role of Gal-8, the co-stimulatory activity of borderline T-cell-specific responses allows the proposal of Gal-8 as an adjuvant in vaccination. To study the relevance of glycan-lectin interaction to these T-cell activities, we generated a double-mutated protein (Gal-8mut) by replacing canonical arginine residues on each CRD, so as to abolish sugar-binding capacity. As expected, Gal-8mut was unable to bind to lactosyl-Sepharose, confirming that lactose recognition was precluded; however, preservation of lectin activity was still evident since Gal-8mut displayed haemoagglutinatory effects and binding capacity to the T-cell surface. To search for glycan affinity, a glycan microarray analysis was conducted which revealed that Gal-8mut lost most low- and intermediate-, but retained high-, affinity interactions, mainly to polylactosamines and blood group antigens. These findings were supported further by molecular modelling. Regarding biological activity, Gal-8mut was unable to induce T-cell proliferation, but efficiently co-stimulated antigen-specific responses, bothin vitroandin vivo.Therefore Gal-8mut represents a useful tool to dissect the specificities of lectin-glycan interactions underlying distinctive Gal-8 activities on T-cell biology. Moreover, given its distinguishing properties, Gal-8mut could be used to enhance borderline immune responses without the non-specific pro-inflammatory activity or other potential adverse effects. PMID:26795039

  9. Bilateral Administration of Autologous CD133+ Cells in Ambulatory Patients with Refractory Critical Limb Ischemia: Lessons Learned from a Pilot Randomized, Double blind, Placebo-controlled Trial

    PubMed Central

    Raval, Amish N.; Schmuck, Eric; Tefera, Girma; Leitzke, Cathlyn; Ark, Cassondra Vander; Hei, Derek; Centanni, John M.; de Silva, Ranil; Koch, Jill; Chappell, Richard; Hematti, Peiman

    2014-01-01

    Introduction CD133+ cells confer angiogenic potential and may be beneficial for the treatment of critical limb ischemia (CLI). However, patient selection, blinding methods and endpoints for clinical trials is challenging. We hypothesized that bilateral intramuscular administration of cytokine mobilized CD133+ cells in ambulatory patients with refractory CLI would be feasible and safe. Methods In this double-blind, randomized, sham-controlled trial, subjects received subcutaneous injections of granulocyte colony stimulating factor (10 mcg/kg/d) for 5 days, followed by leukapheresis, and intramuscular administration of 50-400 million sorted CD133+ cells delivered into both legs. Control subjects received normal saline injections, sham leukapheresis and intramuscular injection of placebo buffered solution. Subjects were followed for 1 year. An aliquot of CD133+ cells was collected from each subject to test for genes associated with cell senescence. Results 70 subjects were screened, of whom 10 were eligible. Subject enrollment was suspended due to a high rate of mobilization failure in subjects randomized to treatment. Of 10 subjects enrolled (7 randomized to treatment, 3 randomized to control), there were no differences in serious adverse events at 12 months and blinding was preserved. There were non-significant trends toward improved amputation free survival, 6 minute walk distance, walking impairment questionnaire and quality of life in subjects randomized to treatment. Successful CD133+ mobilizers expressed fewer senescence associated genes compared to poor mobilizers. Conclusion Bilateral administration of autologous CD133+ cell in ambulatory CLI subjects was safe and blinding was preserved. However, poor mobilization efficiency combined with high CD133+ senescence suggests futility in this approach. PMID:25239491

  10. Study of Method for Designing the Power and the Capacitance of Fuel Cells and Electric Double-Layer Capacitors of Hybrid Railway Vehicle

    NASA Astrophysics Data System (ADS)

    Takizawa, Kenji; Kondo, Keiichiro

    A hybrid railway traction system with fuel cells (FCs) and electric double layer-capacitors (EDLCs) is discussed in this paper. This system can save FC costs and absorb the regenerative energy. A method for designing FCs and EDLCs on the basis of the output power and capacitance, respectively, has not been reported, even though their design is one of the most important technical issues encountered in the design of hybrid railway vehicles. Such design method is presented along with a train load profile and an energy management strategy. The design results obtained using the proposed method are verified by performing numerical simulations of a running train. These results reveal that the proposed method for designing the EDLCs and FCs on the basis of the capacitance and power, respectively, and by using a method for controlling the EDLC voltage is sufficiently effective in designing efficient EDLCs and FCs of hybrid railway traction systems.

  11. Characterization and evaluation of double perovskites LnBaCoFeO5+δ (Ln = Pr and Nd) as intermediate-temperature solid oxide fuel cell cathodes

    NASA Astrophysics Data System (ADS)

    Jin, Fangjun; Xu, Huawei; Long, Wen; Shen, Yu; He, Tianmin

    2013-12-01

    Double perovskites LnBaCoFeO5+δ (Ln = Pr and Nd, PBCF and NBCF) are comparatively investigated as potential cathode materials for intermediate-temperature solid oxide fuel cells (IT-SOFCs). LnBaCoFeO5+δ materials are chemically compatible with La0.9Sr0.1Ga0.8Mg0.2O3-δ (LSGM) electrolyte at temperatures below 1000 °C. Fe and Co ions in LnBaCoFeO5+δ exist in two oxidation states, 3+ and 4+. Pr ions are found in PBCF mostly as Pr3+. Thermal expansion coefficients (TECs) of PBCF and NBCF are 21.0 × 10-6 and 19.5 × 10-6 K-1, respectively, between 30 and 1000 °C; these are lower than the TECs of undoped LnBaCo2O5+δ. The best electrical conductivity for both materials is observed near 350 °C: 321 and 172 S cm-1 for PBCF and NBCF, respectively. Polarization resistances of PBCF and NBCF cathodes on LSGM electrolyte are 0.049 and 0.062 Ω cm2 at 800 °C, respectively. Maximum power densities of the single-cell with Ni/SDC as anode on a 0.3 mm-thick LSGM electrolyte reach 749 and 669 mW cm-2 for PBCF and NBCF cathodes at 800 °C, respectively. As cathodes for application in IT-SOFCs, the performance of PBCF and NBCF double perovskites is promising.

  12. Effusive molecular beam-sampled Knudsen flow reactor coupled to vacuum ultraviolet single photon ionization mass spectrometry using an external free radical source

    SciTech Connect

    Leplat, N.; Rossi, M. J.

    2013-11-15

    A new apparatus using vacuum ultraviolet single photon ionization mass spectrometry (VUV SPIMS) of an effusive molecular beam emanating from a Knudsen flow reactor is described. It was designed to study free radical-molecule kinetics over a significant temperature range (300–630 K). Its salient features are: (1) external free radical source, (2) counterpropagating molecular beam and diffuse VUV photon beam meeting in a crossed-beam ion source of a quadrupole mass spectrometer with perpendicular ion extraction, (3) analog detection of the photocurrent of the free radical molecular cation, and (4) possibility of detecting both free radicals and closed shell species in the same apparatus and under identical reaction conditions owing to the presence of photoelectrons generated by the photoelectric effect of the used VUV-photons. The measured thermal molecular beam-to-background ratio was 6.35 ± 0.39 for Ar and 10.86 ± 1.59 for i-C{sub 4}H{sub 10} at 300 K, a factor of 2.52 and 1.50 smaller, respectively, than predicted from basic gas-dynamic considerations. Operating parameters as well as the performance of key elements of the instrument are presented and discussed. Coupled to an external free radical source a steady-state specific exit flow of 1.6 × 10{sup 11} and 5.0 × 10{sup 11} molecule s{sup −1} cm{sup −3} of C{sub 2}H{sub 5}{sup •} (ethyl) and t-C{sub 4}H{sub 9}{sup •} (t-butyl) free radicals have been detected using VUV SPIMS at their molecular ion m/z 29 and 57, respectively, at 300 K.

  13. Effusive molecular beam-sampled Knudsen flow reactor coupled to vacuum ultraviolet single photon ionization mass spectrometry using an external free radical source.

    PubMed

    Leplat, N; Rossi, M J

    2013-11-01

    A new apparatus using vacuum ultraviolet single photon ionization mass spectrometry (VUV SPIMS) of an effusive molecular beam emanating from a Knudsen flow reactor is described. It was designed to study free radical-molecule kinetics over a significant temperature range (300-630 K). Its salient features are: (1) external free radical source, (2) counterpropagating molecular beam and diffuse VUV photon beam meeting in a crossed-beam ion source of a quadrupole mass spectrometer with perpendicular ion extraction, (3) analog detection of the photocurrent of the free radical molecular cation, and (4) possibility of detecting both free radicals and closed shell species in the same apparatus and under identical reaction conditions owing to the presence of photoelectrons generated by the photoelectric effect of the used VUV-photons. The measured thermal molecular beam-to-background ratio was 6.35 ± 0.39 for Ar and 10.86 ± 1.59 for i-C4H10 at 300 K, a factor of 2.52 and 1.50 smaller, respectively, than predicted from basic gas-dynamic considerations. Operating parameters as well as the performance of key elements of the instrument are presented and discussed. Coupled to an external free radical source a steady-state specific exit flow of 1.6 × 10(11) and 5.0 × 10(11) molecule s(-1) cm(-3) of C2H5(●) (ethyl) and t-C4H9(●) (t-butyl) free radicals have been detected using VUV SPIMS at their molecular ion m/z 29 and 57, respectively, at 300 K. PMID:24289411

  14. Xenopus Cds1 Is Regulated by DNA-Dependent Protein Kinase and ATR during the Cell Cycle Checkpoint Response to Double-Stranded DNA Ends

    PubMed Central

    McSherry, Troy D.; Mueller, Paul R.

    2004-01-01

    The checkpoint kinase Cds1 (Chk2) plays a key role in cell cycle checkpoint responses with functions in cell cycle arrest, DNA repair, and induction of apoptosis. Proper regulation of Cds1 is essential for appropriate cellular responses to checkpoint-inducing insults. While the kinase ATM has been shown to be important in the regulation of human Cds1 (hCds1), here we report that the kinases ATR and DNA-dependent protein kinase (DNA-PK) play more significant roles in the regulation of Xenopus Cds1 (XCds1). Under normal cell cycle conditions, nonactivated XCds1 constitutively associates with a Xenopus ATR complex. The association of XCds1 with this complex does not require a functional forkhead activation domain but does require a putative SH3 binding region that is found in XCds1. In response to double-stranded DNA ends, the amino terminus of XCds1 is rapidly phosphorylated in a sequential pattern. First DNA-PK phosphorylates serine 39, a site not previously recognized as important in Cds1 regulation. Xenopus ATM, ATR, and/or DNA-PK then phosphorylate three consensus serine/glutamine sites. Together, these phosphorylations have the dual function of inducing dissociation from the ATR complex and independently promoting the full activation of XCds1. Thus, the checkpoint-mediated activation of XCds1 requires phosphorylation by multiple phosphoinositide 3-kinase-related kinases, protein-protein dissociation, and autophosphorylation. PMID:15509799

  15. Uncoupling and reactive oxygen species (ROS)--a double-edged sword for β-cell function? "Moderation in all things".

    PubMed

    Collins, Sheila; Pi, Jingbo; Yehuda-Shnaidman, Einav

    2012-12-01

    The ability of the mitochondrion to (a) manage fuel import to oxidize for adenosine tri-phosphate (ATP) generation while (b) protecting itself and the cellular environment from electron leak, which can generate highly reactive oxygen species (ROS) is a delicate balancing act. ATP is the currency of the cell and as such serves a signaling function as a substrate partner to many kinases and ion channels. While various ROS species have been viewed as a dangerous and toxic group of molecules, it also has a role as a signal derived from mitochondria, as well as other enzymatic sources: a double-edged sword. Current efforts to understand the biochemical mechanisms affected by ROS as a signal--usually noted to be hydrogen peroxide (H(2)O(2))--are exciting, but this duality of ROS effects also pose challenges in managing its levels to protect cells. The mitochondrial uncoupling protein-2 (UCP2), UCP3, and the permeability transition pore have been integral to efforts to try to understand what role mitochondrial-derived ROS have in cells. In this piece we reflect on mitochondrial ROS and uncoupling proteins as signaling regulators. It seems that when it comes to ROS and uncoupling the proverb "Moderation in all things" is apt. PMID:23168277

  16. Meiotic recombination initiated by a double-strand break in rad50{Delta} yeast cells otherwise unable to initiate meiotic recombination

    SciTech Connect

    Malkova, A.; Haber, J.E.; Dawson, D.

    1996-06-01

    Meiotic recombination in Saccharomyces cerevisiae is initiated by double-strand breaks (DSBs). We have developed a system to compare the properties of meiotic DSBs with those created by the site-specific HO endonuclease. HO endonuclease was expressed under the control of the meiotic-specific SPO13 promoter, creating a DSB at a single site on one of yeast`s 16 chromosomes. In Rad{sup +} strains the times of appearance of the HO-induced DSBs and of subsequent recombinants are coincident with those induced by normal meiotic DSBs. Physical monitoring of DNA showed that SPO13::HO induced gene conversions both in Rad{sup +} and in rad50{Delta} cells that cannot initiate normal meiotic DSBs. We find that the RAD50 gene is important, but not essential, for recombination even after a DSB has been created in a meiotic cell. In rad50{Delta} cells, some DSBs are not repaired until a broken chromosome has been packaged into a spore and is subsequently germinated. This suggests that a broken chromosome does not signal an arrest of progression through meiosis. The recombination defect in rad50{Delta} diploids is not, however, meiotic specific, as mitotic rad50 diploids, experiencing an HO-induced DSB, exhibit similar departures from wild-type recombination. 57 refs., 5 figs., 3 tabs.

  17. The effects of cell assembly compression on the performance of carbon electrochemical double-layer capacitor electrodes

    NASA Astrophysics Data System (ADS)

    Gourdin, Gerald; Jiang, Thomas; Smith, Patricia; Qu, Deyang

    2012-10-01

    Our previous work concluded that the application of force altered the physical structure of the activated carbon electrodes, which resulted in a decrease in the accessible surface area and a displacement of the electrolyte. In this work, the response that different carbon material electrodes exhibit to an applied force was evaluated. Activated carbon powders possess different porous structures, which would exhibit different behaviors when subjected to an applied force and after the release of that force. Cyclic voltammetry and electrochemical impedance spectroscopy (EIS) were used to characterize the response behaviors of the different carbons. Furthermore, a porosimetry analysis was conducted on the carbon material of the electrode before and after the application of force. It was concluded that the application of force shifted the pore distribution toward overall smaller pores through a compression of the porous structure of the carbon. This resulted in a decrease in the more easily accessible surface area, which was exhibited as a decrease in the capacitance values as calculated from the cyclic voltammetry data. There was no longer sufficient time to access the now smaller powers at the given time scale of the cyclic voltammetry analysis, which negatively impacted the formation of the double layer.

  18. Cholesterol interactions with tetracosenoic acid phospholipids in model cell membranes: role of the double-bond position.

    PubMed

    Ayanoglu, E; Chiche, B H; Beatty, M; Djerassi, C; Düzgüneş, N

    1990-04-10

    The synthesis and thermotropic properties of 1,2-di-(9Z)-9-tetracosenoylphosphatidylcholine [delta 9-PC(24:1,24:1), 1], 1,2-di-(5Z)-5-tetracosenoylphosphatidylcholine [delta 5-PC(24:1,24:1), 2], and 1,2-di-(15Z)-15- tetracosenoylphosphatidylcholine [delta 15-PC(24:1,24:1), 3] are reported. Liposomes prepared from these phospholipids differ from those of the natural sponge phospholipids, 1,2-di-(5Z,9Z)-5,9-hexacosadienoylphosphatidylcholine (4a) and the corresponding ethanolamine (4b), both of which virtually exclude cholesterol from their bilayers. The behavior of 1 and 2 is similar to that of 1,2-di-(6Z,9Z)-6,9-hexacosadienoylphosphatidylcholine (5), which exhibits a partial molecular interaction with cholesterol. In the case of 3, cholesterol appears to interact with the saturated acyl chain regions of this phospholipid in a manner similar to that of its interaction with DPPC acyl chains. This study delineates the effect of the double-bond location in long fatty acyl chains of phospholipids on their interactions with cholesterol. PMID:2354147

  19. Ciprofloxacin-induced G2 arrest and apoptosis in TK6 lymphoblastoid cells is not dependent on DNA double-strand break formation

    PubMed Central

    Smart, Daniel J.; Halicka, H. Dorota; Traganos, Frank; Darzynkiewicz, Zbigniew; Williams, Gary M.

    2008-01-01

    Drugs developed for the treatment of conditions other than neoplasia can also show promise as potential antitumor agents. The fluoroquinolone antibiotic ciprofloxacin (CPFX) is known to modulate cycle cell progression and apoptosis in cancer cells, and is thought to induce DNA double-strand breaks (DSBs) via topoisomerase II (topo II) inhibition and stabilized cleavage complex (SCC) formation. DSBs trigger Ser-139 phosphorylation of histone H2AX (γH2AX) by PI-3-like kinases including ATM; γH2AX can serve as a marker of DNA damage when measured in situ using immunocytochemistry and flow cytometry. The aim of the present study was to investigate the relationship between CPFX-mediated DNA damage and induction of apoptosis in human lymphoblastoid cells and phytohaemagglutinin (PHA)-stimulated lymphocytes (Lymphs). Treatment of TK6 cells (wild-type p53) with 100 µg/ml CPFX for 2-10 h produced no increase in γH2AX; to the contrary, its level in S phase cells was reduced at 10 h compared to controls. Nevertheless, stabilization of topo IIα, ATM Ser-1981 phosphorylation and G2 arrest was observed in TK6 cells exposed to CPFX for ≥4 h. However, following 24 h treatment, γH2AX was dramatically increased in a sub-population of cells indicating the onset of apoptosis (confirmed by presence of activated caspase 3). CPFX had a similar lack of effect on induction of γH2AX at early time points in WTK1 and NH32 cells (devoid of functional p53) and proliferating Lymphs, however, induction of apoptosis was less pronounced than in TK6 cells. Formation of SCC and activation of ATM (but lack of γH2AX induction) indicates topo II-mediated chromatin or DNA changes in the absence of DSBs; ATM activation apparently triggers the G2M checkpoint leading to G2 arrest. The subsequent induction of apoptosis appears to be facilitated by functional p53. CPFX may therefore have a potential use as a chemotherapeutic agent in the treatment of lymphoblast-derived cancer. PMID:18059176

  20. Gefitinib Radiosensitizes Stem-Like Glioma Cells: Inhibition of Epidermal Growth Factor Receptor-Akt-DNA-PK Signaling, Accompanied by Inhibition of DNA Double-Strand Break Repair

    SciTech Connect

    Kang, Khong Bee; Zhu Congju; Wong Yinling; Gao Qiuhan; Ty, Albert; Wong, Meng Cheong

    2012-05-01

    Purpose: We compared radiosensitivity of brain tumor stem cells (BTSCs) with matched nonstem glioma cells, and determined whether gefitinib enhanced BTSC radiosensitivity by inhibiting epidermal growth factor receptor (EGFR)-Akt-DNA-dependent protein kinase (DNA-PK) signaling, followed by enhanced DNA double-stand breaks (DSBs) and inhibition of DSB repair. Methods and Materials: Radiosensitivity of stem-like gliomaspheres and nonstem glioma cells (obtained at patient neurosurgical resection) were evaluated by clonogenic assays, {gamma}-H{sub 2}AX immunostaining and cell cycle distribution. Survival of irradiated and nonirradiated NOD-SCID mice intracranially implanted with stem-like gliomaspheres were monitored. Glioma cells treated with gefitinib, irradiation, or both were assayed for clonogenic survival, {gamma}-H{sub 2}AX immunostaining, DNA-PKcs expression, and phosphorylation of EGFR and Akt. Results: Stem-like gliomaspheres displayed BTSC characteristics of self-renewal; differentiation into lineages of neurons, oligodendrocytes, and astrocytes; and initiation of glioma growth in NOD-SCID mice. Irradiation dose-dependently reduced clonogenic survival, induced G{sub 2}/M arrest and increased {gamma}-H{sub 2}AX immunostaining of nonstem glioma cells, but not stem-like gliomaspheres. There was no difference in survival of irradiated and nonirradiated mice implanted with stem-like gliomaspheres. The addition of gefitinib significantly inhibited clonogenic survival, increased {gamma}-H{sub 2}AX immunostaining, and reduced DNA-PKcs expression of irradiated stem-like gliomaspheres, without affecting irradiated-nonstem glioma cells. Gefitinib alone, and when combined with irradiation, inhibited phosphorylation of EGFR (Y1068 and Y1045) and Akt (S473) in stem-like gliomaspheres. In nonstem glioma cells, gefitinib alone inhibited EGFR Y1068 phosphorylation, with further inhibition by combined gefitinib and irradiation. Conclusions: Stem-like gliomaspheres are

  1. GC-Rich Extracellular DNA Induces Oxidative Stress, Double-Strand DNA Breaks, and DNA Damage Response in Human Adipose-Derived Mesenchymal Stem Cells

    PubMed Central

    Kostyuk, Svetlana; Smirnova, Tatiana; Kameneva, Larisa; Porokhovnik, Lev; Speranskij, Anatolij; Ershova, Elizaveta; Stukalov, Sergey; Izevskaya, Vera; Veiko, Natalia

    2015-01-01

    Background. Cell free DNA (cfDNA) circulates throughout the bloodstream of both healthy people and patients with various diseases. CfDNA is substantially enriched in its GC-content as compared with human genomic DNA. Principal Findings. Exposure of haMSCs to GC-DNA induces short-term oxidative stress (determined with H2DCFH-DA) and results in both single- and double-strand DNA breaks (comet assay and γH2AX, foci). As a result in the cells significantly increases the expression of repair genes (BRCA1 (RT-PCR), PCNA (FACS)) and antiapoptotic genes (BCL2 (RT-PCR and FACS), BCL2A1, BCL2L1, BIRC3, and BIRC2 (RT-PCR)). Under the action of GC-DNA the potential of mitochondria was increased. Here we show that GC-rich extracellular DNA stimulates adipocyte differentiation of human adipose-derived mesenchymal stem cells (haMSCs). Exposure to GC-DNA leads to an increase in the level of RNAPPARG2 and LPL (RT-PCR), in the level of fatty acid binding protein FABP4 (FACS analysis) and in the level of fat (Oil Red O). Conclusions. GC-rich fragments in the pool of cfDNA can potentially induce oxidative stress and DNA damage response and affect the direction of mesenchymal stem cells differentiation in human adipose—derived mesenchymal stem cells. Such a response may be one of the causes of obesity or osteoporosis. PMID:26273425

  2. To Nick or Not to Nick: Comparison of I-SceI Single- and Double-Strand Break-Induced Recombination in Yeast and Human Cells

    PubMed Central

    Katz, Samantha S.; Gimble, Frederick S.; Storici, Francesca

    2014-01-01

    Genetic modification of a chromosomal locus to replace an existing dysfunctional allele with a corrected sequence can be accomplished through targeted gene correction using the cell's homologous recombination (HR) machinery. Gene targeting is stimulated by generation of a DNA double-strand break (DSB) at or near the site of correction, but repair of the break via non-homologous end-joining without using the homologous template can lead to deleterious genomic changes such as in/del mutations, or chromosomal rearrangements. By contrast, generation of a DNA single-strand break (SSB), or nick, can stimulate gene correction without the problems of DSB repair because the uncut DNA strand acts as a template to permit healing without alteration of genetic material. Here, we examine the ability of a nicking variant of the I-SceI endonuclease (K223I I-SceI) to stimulate gene targeting in yeast Saccharomyces cerevisiae and in human embryonic kidney (HEK-293) cells. K223I I-SceI is proficient in both yeast and human cells and promotes gene correction up to 12-fold. We show that K223I I-SceI-driven recombination follows a different mechanism than wild-type I-SceI-driven recombination, thus indicating that the initial DNA break that stimulates recombination is not a low-level DSB but a nick. We also demonstrate that K223I I-SceI efficiently elevates gene targeting at loci distant from the break site in yeast cells. These findings establish the capability of the I-SceI nickase to enhance recombination in yeast and human cells, strengthening the notion that nicking enzymes could be effective tools in gene correction strategies for applications in molecular biology, biotechnology, and gene therapy. PMID:24558436

  3. PI3K-delta mediates double-stranded RNA-induced upregulation of B7-H1 in BEAS-2B airway epithelial cells

    SciTech Connect

    Kan-o, Keiko; Matsumoto, Koichiro; Asai-Tajiri, Yukari; Fukuyama, Satoru; Hamano, Saaka; Seki, Nanae; Nakanishi, Yoichi; Inoue, Hiromasa

    2013-05-31

    Highlights: •Double-stranded RNA upregulates B7-H1 on BEAS-2B airway epithelial cells. •The upregulation of B7-H1 is attenuated by inhibition of PI3Kδ isoform. •PI3Kδ-mediated upregulation of B7-H1 is independent of NF-κB activation. •Inhibition of PI3Kδ may prevent persistent viral infection induced by B7-H1. -- Abstract: Airway viral infection disturbs the health-related quality of life. B7-H1 (also known as PD-L1) is a coinhibitory molecule associated with the escape of viruses from the mucosal immunity, leading to persistent infection. Most respiratory viruses generate double-stranded (ds) RNA during replication. The stimulation of cultured airway epithelial cells with an analog of viral dsRNA, polyinosinic-polycytidylic acid (poly IC) upregulates the expression of B7-H1 via activation of the nuclear factor κB(NF-κB). The mechanism of upregulation was investigated in association with phosphatidylinositol 3-kinases (PI3Ks). Poly IC-induced upregulation of B7-H1 was profoundly suppressed by a pan-PI3K inhibitor and partially by an inhibitor or a small interfering (si)RNA for PI3Kδ in BEAS-2B cells. Similar results were observed in the respiratory syncytial virus-infected cells. The expression of p110δ was detected by Western blot and suppressed by pretreatment with PI3Kδ siRNA. The activation of PI3Kδ is typically induced by oxidative stress. The generation of reactive oxygen species was increased by poly IC. Poly IC-induced upregulation of B7-H1 was attenuated by N-acetyl-L-cysteine, an antioxidant, or by oxypurinol, an inhibitor of xanthine oxidase. Poly IC-induced activation of NF-κB was suppressed by a pan-PI3K inhibitor but not by a PI3Kδ inhibitor. These results suggest that PI3Kδ mediates dsRNA-induced upregulation of B7-H1 without affecting the activation of NF-κB.

  4. Investigation of a redox-sensitive predictive model of mouse embryonic stem cells differentiation using quantitative nuclease protection assays and glutathione redox status

    EPA Science Inventory

    Investigation of a redox-sensitive predictive model of mouse embryonic stem cell differentiation via quantitative nuclease protection assays and glutathione redox status Chandler KJ,Hansen JM, Knudsen T,and Hunter ES 1. U.S. Environmental Protection Agency, Research Triangl...

  5. DNA double-strand break repair in parental chromatin of mouse zygotes, the first cell cycle as an origin of de novo mutation.

    PubMed

    Derijck, Alwin; van der Heijden, Godfried; Giele, Maud; Philippens, Marielle; de Boer, Peter

    2008-07-01

    In the human, the contribution of the sexes to the genetic load is dissimilar. Especially for point mutations, expanded simple tandem repeats and structural chromosome mutations, the contribution of the male germline is dominant. Far less is known about the male germ cell stage(s) that are most vulnerable to mutation contraction. For the understanding of de novo mutation induction in the germline, mechanistic insight of DNA repair in the zygote is mandatory. At the onset of embryonic development, the parental chromatin sets occupy one pronucleus (PN) each and DNA repair can be regarded as a maternal trait, depending on proteins and mRNAs provided by the oocyte. Repair of DNA double-strand breaks (DSBs) is executed by non-homologous end joining (NHEJ) and homologous recombination (HR). Differentiated somatic cells often resolve DSBs by NHEJ, whereas embryonic stem cells preferably use HR. We show NHEJ and HR to be both functional during the zygotic cell cycle. NHEJ is already active during replacement of sperm protamines by nucleosomes. The kinetics of G1 repair is influenced by DNA-PK(cs) hypomorphic activity. Both HR and NHEJ are operative in S-phase, HR being more active in the male PN. DNA-PK(cs) deficiency upregulates the HR activity. Both after sperm remodeling and at first mitosis, spontaneous levels of gammaH2AX foci (marker for DSBs) are high. All immunoflurescent indices of DNA damage and DNA repair point at greater spontaneous damage and induced repair activity in paternal chromatin in the zygote. PMID:18353795

  6. Induction and Rejoining of DNA Double Strand Breaks Assessed by H2AX Phosphorylation in Melanoma Cells Irradiated with Proton and Lithium Beams

    SciTech Connect

    Ibanez, Irene L.; Bracalente, Candelaria; Molinari, Beatriz L.; Palmieri, Monica A.; Policastro, Lucia; Kreiner, Andres J.; Burlon, Alejandro A.; Valda, Alejandro; Navalesi, Daniela; Davidson, Jorge; Davidson, Miguel; Vazquez, Monica; Ozafran, Mabel; Duran, Hebe

    2009-07-15

    Purpose: The aim of this study was to evaluate the induction and rejoining of DNA double strand breaks (DSBs) in melanoma cells exposed to low and high linear energy transfer (LET) radiation. Methods and Materials: DSBs and survival were determined as a function of dose in melanoma cells (B16-F0) irradiated with monoenergetic proton and lithium beams and with a gamma source. Survival curves were obtained by clonogenic assay and fitted to the linear-quadratic model. DSBs were evaluated by the detection of phosphorylated histone H2AX ({gamma}H2AX) foci at 30 min and 6 h post-irradiation. Results: Survival curves showed the increasing effectiveness of radiation as a function of LET. {gamma}H2AX labeling showed an increase in the number of foci vs. dose for all the radiations evaluated. A decrease in the number of foci was found at 6 h post-irradiation for low LET radiation, revealing the repair capacity of DSBs. An increase in the size of {gamma}H2AX foci in cells irradiated with lithium beams was found, as compared with gamma and proton irradiations, which could be attributed to the clusters of DSBs induced by high LET radiation. Foci size increased at 6 h post-irradiation for lithium and proton irradiations in relation with persistent DSBs, showing a correlation with surviving fraction. Conclusions: Our results showed the response of B16-F0 cells to charged particle beams evaluated by the detection of {gamma}H2AX foci. We conclude that {gamma}H2AX foci size is an accurate parameter to correlate the rejoining of DSBs induced by different LET radiations and radiosensitivity.

  7. Double Methotrexate-Modified Neuropeptide Y Analogues Express Increased Toxicity and Overcome Drug Resistance in Breast Cancer Cells.

    PubMed

    Böhme, David; Krieghoff, Jan; Beck-Sickinger, Annette G

    2016-04-14

    Bioconjugates containing the neuropeptide Y (NPY) analogue [F(7),P(34)]-NPY as targeting moiety are able to deliver toxic agents specifically to breast cancer cells that overexpress the human Y1-receptor (hY1R). To increase their activity, multiple toxophores can be attached to one peptide. Herein, synthesis and characterization of [F(7),P(34)]-NPY conjugates containing two methotrexate (MTX) molecules are presented. First, carboxytetramethylrhodamine was linked to [F(7),P(34)]-NPY by amide or enzymatic linkage. The conjugate containing the enzymatic cleavage site showed high extracellular stability and fast intracellular release. Then, MTX was introduced at positions four and 22 of [F(7),P(34)]-NPY, connected by enzymatic or amide linkage. The toxicity of the analogues on breast cancer cells was hY1R-mediated and dependent on the used linkage and amount of toxophores. Furthermore, conjugates revealed higher potency than MTX on MTX-resistant cells. These results emphasize that peptide-drug conjugates can overcome drug resistance and that the attachment of multiple cleavable toxophores enhances the efficiency of this smart delivery system. PMID:26985967

  8. Cell nonhomologous end joining capacity controls SAF-A phosphorylation by DNA-PK in response to DNA double-strand breaks inducers.

    PubMed

    Britton, Sébastien; Froment, Carine; Frit, Philippe; Monsarrat, Bernard; Salles, Bernard; Calsou, Patrick

    2009-11-15

    Aiming to identify novel phosphorylation sites in response to DNA double-strand breaks (DSB) inducers, we have isolated a phosphorylation site on KU70. Unexpectedly, a rabbit antiserum raised against this site cross-reacted with a 120 kDa protein in cells treated by DNA DSB inducers. We identified this protein as SAF-A/hnRNP U, an abundant and essential nuclear protein containing regions binding DNA or RNA. The phosphorylation site was mapped at S59 position in a sequence context favoring a "S-hydrophobic" consensus model for DNA-PK phosphorylation site in vivo. This site was exclusively phosphorylated by DNA-PK in response to DNA DSB inducers. In addition, the extent and duration of this phosphorylation was in inverse correlation with the capacity of the cells to repair DSB by Nonhomologous End Joining. These results bring a new link between the hnRNP family and the DNA damage response. Addtionaly, the mapped phospho-site on SAF-A might serve as a potential bio-marker for DNA-PK activity in academic studies and clinical analyses of DNA-PK activators or inhibitors. PMID:19844162

  9. ZnO double layer film with a novel organic sensitizer as an efficient photoelectrode for dye-sensitized solar cells

    NASA Astrophysics Data System (ADS)

    Lee, Chuan-Pei; Chen, Ping-Wei; Li, Chun-Ting; Huang, Yi-June; Li, Sie-Rong; Chang, Ling-Yu; Chen, Pei-Yu; Lin, Lu-Yin; Vittal, R.; Sun, Shih-Sheng; Lin, Jiang-Jen; Ho, Kuo-Chuan

    2016-09-01

    A novel organic sensitizer, coded CR147, is applied to sensitize a ZnO-based dye-sensitized solar cell (DSSC). The common problem of ZnO dissolution and Zn2+/dye agglomeration, caused by the use of ruthenium-based dyes, is solved by the application of this CR147 dye. The highest power conversion efficiency (η) of 4.77% is achieved for the DSSC using a photoanode with a film of commercial ZnO nanoparticles (Csbnd ZnO) and the CR147 dye, while the η is only 3.41% for the DSSC with the commercial N719 dye. The cell performance with the CR147 dye is further improved by using a photoanode with the double layer ZnO film (D-ZnO), composed of an underlayer with coral-like ZnO nanocrystals and an overlayer with hexagonal club-like ZnO submicrocrystals. The DSSC with the D-ZnO film exhibits an η of 6.89%, which is ca. 45% higher than that of the DSSC with the Csbnd ZnO film (4.77%). This higher efficiency is attributed to the superior charge transfer and light-scattering abilities provided by coral-like ZnO nanocrystals and hexagonal clubs-like ZnO submicrocrystals, respectively, with reference to these parameters of Csbnd ZnO.

  10. Non-coding Double-stranded RNA and Antimicrobial Peptide LL-37 Induce Growth Factor Expression from Keratinocytes and Endothelial Cells.

    PubMed

    Adase, Christopher A; Borkowski, Andrew W; Zhang, Ling-Juan; Williams, Michael R; Sato, Emi; Sanford, James A; Gallo, Richard L

    2016-05-27

    A critical function for skin is that when damaged it must simultaneously identify the nature of the injury, repair barrier function, and limit the intrusion of pathogenic organisms. These needs are carried out through the detection of damage-associated molecular patterns (DAMPs) and a response that includes secretion of cytokines, chemokines, growth factors, and antimicrobial peptides (AMPs). In this study, we analyzed how non-coding double-stranded RNA (dsRNAs) act as a DAMP in the skin and how the human cathelicidin AMP LL-37 might influence growth factor production in response to this DAMP. dsRNA alone significantly increased the expression of multiple growth factors in keratinocytes, endothelial cells, and fibroblasts. Furthermore, RNA sequencing transcriptome analysis found that multiple growth factors increase when cells are exposed to both LL-37 and dsRNA, a condition that mimics normal wounding. Quantitative PCR and/or ELISA validated that growth factors expressed by keratinocytes in these conditions included, but were not limited to, basic fibroblast growth factor (FGF2), heparin-binding EGF-like growth factor (HBEGF), vascular endothelial growth factor C (VEGFC), betacellulin (BTC), EGF, epiregulin (EREG), and other members of the transforming growth factor β superfamily. These results identify a novel role for DAMPs and AMPs in the stimulation of repair and highlight the complex interactions involved in the wound environment. PMID:27048655

  11. New epifluorescence microscope providing pairs of specific fluorescence images of double-stained cells for simultaneous visual perception and for quantification

    NASA Astrophysics Data System (ADS)

    Heiden, Thomas; Tribukait, Bernhard

    1996-05-01

    A new epi-fluorescence microscope for analysis of cells stained with two fluorochromes which can be spectrally isolated is described. The system makes it possible to perform independent and specific spectral selection of each dye (e.g. DAPI and CY3) while perceiving the two specific images simultaneously by eye. The optics uses splitting of the primary excitation and emission light beams, independent modification of the separated beams, and their reunification. Modifications in the separated beams comprise: (1) isolation of specific wavelengths (365 nm and 546 nm in the excitation light path, 435 - 500 nm and 590 - 750 nm in the emission light beams), (2) wavelength switching without image displacement and blur by means of a light chopper alternating between ultraviolet-excitation/blue-detection and green-excitation/red-detection at frequencies of up to 140 Hz for observation by eye without image flicker, and (3) the possible separate positioning of lenses for compensation of chromatic aberrations. The system demonstrates a good transmission of the chosen wavelengths. A high specificity of double fluorescence analysis with minimal effects of spectral overlap was attained with good temporal resolution. It has been shown that it is feasible to obtain separate chromatic compensations for the use of a microscope objective in spectral regions outside the range for which the objective is corrected. Quantitative and independent measurements of the two fluorescence images by a CCD camera synchronized with the light chopper are feasible. In conclusion, this imaging system is outlined for highly specific visual analysis and exact quantitative measurement of double fluorescence labeled specimens in cytology and histology.

  12. Double replacement: strategy for efficient introduction of subtle mutations into the murine Col1a-1 gene by homologous recombination in embryonic stem cells.

    PubMed Central

    Wu, H; Liu, X; Jaenisch, R

    1994-01-01

    A subtle mutation that rendered type I collagen resistant to mammalian collagenase has been introduced into the murine Col1a-1 (recently redesignated Cola-1) gene by homologous recombination in embryonic stem (ES) cells. Initially, a "hit and run" procedure was used. Since two steps were required for introducing each mutation and more than one mutation was to be introduced in the same genomic region independently, we have developed a streamlined procedure that involves two sequential replacement-type homologous recombination events. In the first step, an internal deletion was introduced into the Col1a-1 locus along with the positive and negative selectable markers, neo and tk, to mark the region of interest. G418-resistant homologous recombinants were isolated and used in the second step in which the deleted Col1a-1 allele was replaced with a construct containing the desired mutation. Homologous recombinants containing the mutation were identified among the Tk- ES clones after selection with FIAU [1-(2-deoxy-2-fluoro-beta-D-arabinofuranosyl)-5-iodouracil (called fialuridine)]. Approximately 10% of such clones contained the desired mutation. The double replacement procedure greatly reduces the time and amount of work required to introduce mutations independently into the same or closely linked regions. Once the homologous recombinants derived from the first step are established, the introduction of other mutations into the deleted region becomes a one-step procedure. For X number of introduced mutations, 2X selections are required with the "hit and run" approach, but only X + 1 are required with the double-replacement method. This innovative procedure could be very useful in studies of gene structure and function as well as gene expression and regulation. Images PMID:8146196

  13. Attenuation of experimental colitis in glutathione peroxidase 1 and catalase double knockout mice through enhancing regulatory T cell function.

    PubMed

    Kim, Hyung-Ran; Lee, Anbok; Choi, Eun-Jeong; Kie, Jeong-Hae; Lim, Woosung; Lee, Hyeon Kook; Moon, Byung-In; Seoh, Ju-Young

    2014-01-01

    Reactive oxygen species (ROS) have been implicated in the progression of inflammatory diseases including inflammatory bowel diseases (IBD). Meanwhile, several studies suggested the protective role of ROS in immune-mediated inflammatory diseases, and it was recently reported that dextran sodium sulfate (DSS)-induced colitis was attenuated in mice with an elevated level of ROS due to deficiency of peroxiredoxin II. Regulatory T cells (Tregs) are critical in the prevention of IBD and Treg function was reported to be closely associated with ROS level, but it has been investigated only in lowered levels of ROS so far. In the present study, in order to clarify the relationship between ROS level and Treg function, and their role in the pathogenesis of IBD, we investigated mice with an elevated level of ROS due to deficiency of both glutathione peroxidase (GPx)-1 and catalase (Cat) for the susceptibility of DSS-induced colitis in association with Treg function. The results showed that DSS-induced colitis was attenuated and Tregs were hyperfunctional in GPx1-/- × Cat-/- mice. In vivo administration of N-acetylcysteine (NAC) aggravated DSS-induced colitis and decreased Treg function to the level comparable to WT mice. Attenuated Th17 cell differentiation from naïve CD4+ cells as well as impaired production of IL-6 and IL-17A by splenocytes upon stimulation suggested anti-inflammatory tendency of GPx1-/- × Cat-/- mice. Suppression of Stat3 activation in association with enhancement of indoleamine 2,3-dioxygenase and FoxP3 expression might be involved in the immunosuppressive mechanism of GPx1-/- × Cat-/- mice. Taken together, it is implied that ROS level is critical in the regulation of Treg function, and IBD may be attenuated in appropriately elevated levels of ROS. PMID:24743300

  14. Functional double-shelled silicon nanocrystals for two-photon fluorescence cell imaging: spectral evolution and tuning

    NASA Astrophysics Data System (ADS)

    Chandra, Sourov; Ghosh, Batu; Beaune, Grégory; Nagarajan, Usharani; Yasui, Takao; Nakamura, Jin; Tsuruoka, Tohru; Baba, Yoshinobu; Shirahata, Naoto; Winnik, Françoise M.

    2016-04-01

    Functional near-IR (NIR) emitting nanoparticles (NPs) adapted for two-photon excitation fluorescence cell imaging were obtained starting from octadecyl-terminated silicon nanocrystals (ncSi-OD) of narrow photoluminescence (PL) spectra having no long emission tails, continuously tunable over the 700-1000 nm window, PL quantum yields exceeding 30%, and PL lifetimes of 300 μs or longer. These NPs, consisting of a Pluronic F127 shell and a core made up of assembled ncSi-OD kept apart by an octadecyl (OD) layer, were readily internalized into the cytosol, but not the nucleus, of NIH3T3 cells and were non-toxic. Asymmetrical field-flow fractionation (AF4) analysis was carried out to determine the size of the NPs in water. HiLyte Fluor 750 amine was linked via an amide link to NPs prepared with Pluronic-F127-COOH, as a first demonstration of functional NIR-emitting water dispersible ncSi-based nanoparticles.Functional near-IR (NIR) emitting nanoparticles (NPs) adapted for two-photon excitation fluorescence cell imaging were obtained starting from octadecyl-terminated silicon nanocrystals (ncSi-OD) of narrow photoluminescence (PL) spectra having no long emission tails, continuously tunable over the 700-1000 nm window, PL quantum yields exceeding 30%, and PL lifetimes of 300 μs or longer. These NPs, consisting of a Pluronic F127 shell and a core made up of assembled ncSi-OD kept apart by an octadecyl (OD) layer, were readily internalized into the cytosol, but not the nucleus, of NIH3T3 cells and were non-toxic. Asymmetrical field-flow fractionation (AF4) analysis was carried out to determine the size of the NPs in water. HiLyte Fluor 750 amine was linked via an amide link to NPs prepared with Pluronic-F127-COOH, as a first demonstration of functional NIR-emitting water dispersible ncSi-based nanoparticles. Electronic supplementary information (ESI) available. See DOI: 10.1039/c6nr01437b

  15. Functional double-shelled silicon nanocrystals for two-photon fluorescence cell imaging: spectral evolution and tuning.

    PubMed

    Chandra, Sourov; Ghosh, Batu; Beaune, Grégory; Nagarajan, Usharani; Yasui, Takao; Nakamura, Jin; Tsuruoka, Tohru; Baba, Yoshinobu; Shirahata, Naoto; Winnik, Françoise M

    2016-04-28

    Functional near-IR (NIR) emitting nanoparticles (NPs) adapted for two-photon excitation fluorescence cell imaging were obtained starting from octadecyl-terminated silicon nanocrystals (ncSi-OD) of narrow photoluminescence (PL) spectra having no long emission tails, continuously tunable over the 700-1000 nm window, PL quantum yields exceeding 30%, and PL lifetimes of 300 μs or longer. These NPs, consisting of a Pluronic F127 shell and a core made up of assembled ncSi-OD kept apart by an octadecyl (OD) layer, were readily internalized into the cytosol, but not the nucleus, of NIH3T3 cells and were non-toxic. Asymmetrical field-flow fractionation (AF4) analysis was carried out to determine the size of the NPs in water. HiLyte Fluor 750 amine was linked via an amide link to NPs prepared with Pluronic-F127-COOH, as a first demonstration of functional NIR-emitting water dispersible ncSi-based nanoparticles. PMID:27076260

  16. Mechanism of action studies of lomaiviticin A and the monomeric lomaiviticin aglycon. Selective and potent activity toward DNA double-strand break repair-deficient cell lines.

    PubMed

    Colis, Laureen C; Hegan, Denise C; Kaneko, Miho; Glazer, Peter M; Herzon, Seth B

    2015-05-01

    (-)-Lomaiviticin A (1) and the monomeric lomaiviticin aglycon [aka: (-)-MK7-206, (3)] are cytotoxic agents that induce double-strand breaks (DSBs) in DNA. Here we elucidate the cellular responses to these agents and identify synthetic lethal interactions with specific DNA repair factors. Toward this end, we first characterized the kinetics of DNA damage by 1 and 3 in human chronic myelogenous leukemia (K562) cells. DSBs are rapidly induced by 3, reaching a maximum at 15 min post addition and are resolved within 4 h. By comparison, DSB production by 1 requires 2-4 h to achieve maximal values and >8 h to achieve resolution. As evidenced by an alkaline comet unwinding assay, 3 induces extensive DNA damage, suggesting that the observed DSBs arise from closely spaced single-strand breaks (SSBs). Both 1 and 3 induce ataxia telangiectasia mutated- (ATM-) and DNA-dependent protein kinase- (DNA-PK-) dependent production of phospho-SER139-histone H2AX (γH2AX) and generation of p53 binding protein 1 (53BP1) foci in K562 cells within 1 h of exposure, which is indicative of activation of nonhomologous end joining (NHEJ) and homologous recombination (HR) repair. Both compounds also lead to ataxia telangiectasia and Rad3-related- (ATR-) dependent production of γH2AX at later time points (6 h post addition), which is indicative of replication stress. 3 is also shown to induce apoptosis. In accord with these data, 1 and 3 were found to be synthetic lethal with certain mutations in DNA DSB repair. 1 potently inhibits the growth of breast cancer type 2, early onset- (BRCA2-) deficient V79 Chinese hamster lung fibroblast cell line derivative (VC8), and phosphatase and tensin homologue deleted on chromosome ten- (PTEN-) deficient human glioblastoma (U251) cell lines, with LC50 values of 1.5 ± 0.5 and 2.0 ± 0.6 pM, respectively, and selectivities of >11.6 versus the isogenic cell lines transfected with and expressing functional BRCA2 and PTEN genes. 3 inhibits the growth of the same

  17. Nbs1 ChIP-Seq Identifies Off-Target DNA Double-Strand Breaks Induced by AID in Activated Splenic B Cells

    PubMed Central

    Linehan, Erin K.; Schrader, Carol E.; Stavnezer, Janet

    2015-01-01

    Activation-induced cytidine deaminase (AID) is required for initiation of Ig class switch recombination (CSR) and somatic hypermutation (SHM) of antibody genes during immune responses. AID has also been shown to induce chromosomal translocations, mutations, and DNA double-strand breaks (DSBs) involving non-Ig genes in activated B cells. To determine what makes a DNA site a target for AID-induced DSBs, we identify off-target DSBs induced by AID by performing chromatin immunoprecipitation (ChIP) for Nbs1, a protein that binds DSBs, followed by deep sequencing (ChIP-Seq). We detect and characterize hundreds of off-target AID-dependent DSBs. Two types of tandem repeats are highly enriched within the Nbs1-binding sites: long CA repeats, which can form Z-DNA, and tandem pentamers containing the AID target hotspot WGCW. These tandem repeats are not nearly as enriched at AID-independent DSBs, which we also identified. Msh2, a component of the mismatch repair pathway and important for genome stability, increases off-target DSBs, similar to its effect on Ig switch region DSBs, which are required intermediates during CSR. Most of the off-target DSBs are two-ended, consistent with generation during G1 phase, similar to DSBs in Ig switch regions. However, a minority are one-ended, presumably due to conversion of single-strand breaks to DSBs during replication. One-ended DSBs are repaired by processes involving homologous recombination, including break-induced replication repair, which can lead to genome instability. Off-target DSBs, especially those present during S phase, can lead to chromosomal translocations, deletions and gene amplifications, resulting in the high frequency of B cell lymphomas derived from cells that express or have expressed AID. PMID:26263206

  18. Theoretical study of aromatic-antiaromatic pairs as material in organic solar cells of double light harvesting

    NASA Astrophysics Data System (ADS)

    Martinez, Iván; Schott, Eduardo; Chávez, Ivonne; Manríquez, Juan Manuel; Zarate, Ximena

    2016-08-01

    Molecular light harvesting components of organic solar cells containing antiaromatic and aromatic molecules as organic semiconductors were studied. We found that antiaromatic molecules with indacene core can act as acceptors looking for new options to assemble donor/acceptor interfaces. This is supported by their properties such as molecular orbitals energies, rigid fused core that could promote π-π intermolecular interactions imparting ordered nanostructures, that let high charge mobility thanks to their properly low reorganization energy and the optimum energy offsets of the donor/acceptor interfaces. It was found that pentacene might be an excellent donor and the benzo[g]benz[6,7]indeno[1,2-b]fluorene could act as an acceptor.

  19. A Double-Blind Randomized Phase I Clinical Trial Targeting ALVAC-HIV Vaccine to Human Dendritic Cells

    PubMed Central

    Cox, Josephine H.; Lesho, Emil; Wang, Zhining; Currier, Jeffrey R.; Darden, Janice M.; Polonis, Victoria R.; Vahey, Maryanne T.; Peel, Sheila; Robb, Merlin L.; Michael, Nelson L.; Marovich, Mary A.

    2011-01-01

    Background We conducted a novel pilot study comparing different delivery routes of ALVAC-HIV (vCP205), a canarypox vaccine containing HIV gene inserts: env, gag and pol. We explored the concept that direct ex vivo targeting of human dendritic cells (DC) would enhance the immune response compared to either conventional intramuscular or intradermal injections of the vaccine alone. Methodology/Principal Findings Healthy HIV-1 uninfected volunteers were administered ALVAC-HIV or placebo by intramuscular injection (IM), intradermal injection (ID) or subcutaneous injection (SQ) of autologous ex vivo transfected DC at months 0, 1, 3 and 6. All vaccine delivery routes were well tolerated. Binding antibodies were observed to both the ALVAC vector and HIV-1 gp160 proteins. Modest cellular responses were observed in 2/7 individuals in the DC arm and 1/8 in the IM arm as determined by IFN-γ ELISPOT. Proliferative responses were most frequent in the DC arm where 4/7 individuals had measurable responses to multiple HIV-1 antigens. Loading DC after maturation resulted in lower gene expression, but overall better responses to both HIV-1 and control antigens, and were associated with better IL-2, TNF-α and IFN-γ production. Conclusions/Significance ALVAC-HIV delivered IM, ID or SQ with autologous ex vivo transfected DC proved to be safe. The DC arm was most immunogenic. Proliferative immune responses were readily detected with only modest cytotoxic CD8 T cell responses. Loading mature DC with the live viral vaccine induced stronger immune responses than loading immature DC, despite increased transgene expression with the latter approach. Volunteers who received the autologous vaccine loaded mature DC developed a broader and durable immune response compared to those vaccinated by conventional routes. Trial Registration ClinicalTrials.gov NCT00013572 PMID:21949699

  20. Seeing Double

    NASA Astrophysics Data System (ADS)

    Pesic, Peter

    2003-10-01

    The separateness and connection of individuals is perhaps the central question of human life: What, exactly, is my individuality? To what degree is it unique? To what degree can it be shared, and how? To the many philosophical and literary speculations about these topics over time, modern science has added the curious twist of quantum theory, which requires that the elementary particles of which everything consists have no individuality at all. All aspects of chemistry depend on this lack of individuality, as do many branches of physics. From where, then, does our individuality come? In Seeing Double, Peter Pesic invites readers to explore this intriguing set of questions. He draws on literary and historical examples that open the mind (from Homer to Martin Guerre to Kafka), philosophical analyses that have helped to make our thinking and speech more precise, and scientific work that has enabled us to characterize the phenomena of nature. Though he does not try to be all-inclusive, Pesic presents a broad range of ideas, building toward a specific point of view: that the crux of modern quantum theory is its clash with our ordinary concept of individuality. This represents a departure from the usual understanding of quantum theory. Pesic argues that what is bizarre about quantum theory becomes more intelligible as we reconsider what we mean by individuality and identity in ordinary experience. In turn, quantum identity opens a new perspective on us. Peter Pesic is a Tutor and Musician-in-Residence at St. John's College, Santa Fe, New Mexico. He has a Ph.D. in physics from Stanford University.

  1. Double inflation

    SciTech Connect

    Silk, J.; Turner, M.S.

    1986-04-01

    The Zel'dovich spectrum of adiabatic density perturbations is a generic prediction of inflation. There is increasing evidence that when the spectrum is normalized by observational data on small scales, there is not enough power on large scales to account for the observed large-scale structure in the Universe. Decoupling the spectrum on large and small scales could solve this problem. As a means of decoupling the large and small scales we propose double inflation (i.e., two episodes of inflation). In this scenario the spectrum on large scales is determined by the first episode of inflation and those on small scales by a second episode of inflation. We present three models for such a scenario. By nearly saturating the large angular-scale cosmic microwave anisotropy bound, we can easily account for the observed large-scale structure. We take the perturbations on small scales to be very large, deltarho/rho approx. = 0.1 to 0.01, which results in the production of primordial black holes (PBHs), early formation of structure, reionization of the Universe, and a rich array of astrophysical events. The ..cap omega..-problem is also addressed by our scenario. Allowing the density perturbations produced by the second episode of inflation to be large also lessens the fine-tuning required in the scalar potential and makes reheating much easier. We briefly speculate on the possibility that the second episode of inflation proceeds through the nucleation of bubbles, which today manifest themselves as empty bubbles whose surfaces are covered with galaxies. 37 refs., 1 fig.

  2. CD138 and CD31 Double-Positive Cells Comprise the Functional Antibody-Secreting Plasma Cell Compartment in Primate Bone Marrow

    PubMed Central

    Martinez-Murillo, Paola; Pramanik, Lotta; Sundling, Christopher; Hultenby, Kjell; Wretenberg, Per; Spångberg, Mats; Karlsson Hedestam, Gunilla B.

    2016-01-01

    Plasma cells (PCs) are defined as terminally differentiated B cells that secrete large amounts of immunoglobulin (Ig). PCs that reside in the bone marrow (BM) are responsible for maintaining long-term antibody (Ab) responses after infection and vaccination, while PCs present in the blood are generally short-lived. In rhesus macaques, a species frequently used for the evaluation of human vaccines, B cells resemble those found in humans. However, a detailed characterization of BM-resident rhesus PC phenotype and function is lacking. Here, we examined Ig secretion of distinct rhesus CD138+ populations by B cell ELISpot analysis to couple phenotype with function. We demonstrate that the CD20low/−CD138+CD31+ BM population was highly enriched for antibody-secreting cells with IgG being the predominant isotype (60%), followed by IgA (33%) and IgM (7%). Transmission electron microscopy analysis confirmed PC enrichment in the CD20low/−CD138+CD31+ population with cells containing nuclei with “spokes of a wheel” chromatin structure and prominent rough endoplasmic reticulum. This panel also stained human BM PCs and allowed a clear distinction between BM PCs and short-lived peripheral PCs, providing an improved strategy to isolate PCs from rhesus BM for further analysis. PMID:27446073

  3. Simulation of plasma double-layer structures

    NASA Technical Reports Server (NTRS)

    Borovsky, J. E.; Joyce, G.

    1982-01-01

    Electrostatic plasma double layers are numerically simulated by means of a magnetized 2 1/2 dimensional particle in cell method. The investigation of planar double layers indicates that these one dimensional potential structures are susceptible to periodic disruption by instabilities in the low potential plasmas. Only a slight increase in the double layer thickness with an increase in its obliqueness to the magnetic field is observed. Weak magnetization results in the double layer electric field alignment of accelerated particles and strong magnetization results in their magnetic field alignment. The numerical simulations of spatially periodic two dimensional double layers also exhibit cyclical instability. A morphological invariance in two dimensional double layers with respect to the degree of magnetization implies that the potential structures scale with Debye lengths rather than with gyroradii. Electron beam excited electrostatic electron cyclotron waves and (ion beam driven) solitary waves are present in the plasmas adjacent to the double layers.

  4. Titanium dioxide/silicon hole-blocking selective contact to enable double-heterojunction crystalline silicon-based solar cell

    NASA Astrophysics Data System (ADS)

    Nagamatsu, Ken A.; Avasthi, Sushobhan; Sahasrabudhe, Girija; Man, Gabriel; Jhaveri, Janam; Berg, Alexander H.; Schwartz, Jeffrey; Kahn, Antoine; Wagner, Sigurd; Sturm, James C.

    2015-03-01

    In this work, we use an electron-selective titanium dioxide (TiO2) heterojunction contact to silicon to block minority carrier holes in the silicon from recombining at the cathode contact of a silicon-based photovoltaic device. We present four pieces of evidence demonstrating the beneficial effect of adding the TiO2 hole-blocking layer: reduced dark current, increased open circuit voltage (VOC), increased quantum efficiency at longer wavelengths, and increased stored minority carrier charge under forward bias. The importance of a low rate of recombination of minority carriers at the Si/TiO2 interface for effective blocking of minority carriers is quantitatively described. The anode is made of a poly(3,4-ethylenedioxythiophene):poly(styrenesulfonate) heterojunction to silicon which forms a hole selective contact, so that the entire device is made at a maximum temperature of 100 °C, with no doping gradients or junctions in the silicon. A low rate of recombination of minority carriers at the Si/TiO2 interface is crucial for effective blocking of minority carriers. Such a pair of complementary carrier-selective heterojunctions offers a path towards high-efficiency silicon solar cells using relatively simple and near-room temperature fabrication techniques.

  5. Double primary malignant fibrous histiocytoma and squamous cell carcinoma of the larynx treated with laser laryngeal conservation surgery

    PubMed Central

    Karkos, PD; Dova, S; Sotiriou, S; Markou, K; Kostopoulos, I

    2016-01-01

    Βackground Synchronous multiple malignancies of the larynx are rare. We present a case here of synchronous primary laryngeal squamous cell carcinoma (SCC) and malignant fibrous histiocytoma (MFH) in a patient with hoarseness though with no history of exposure to radiation. Clinical, intraoperative, and histopathological findings in this patient are discussed. Methods Wide laser excision of the left supraglottic lesion and laser cordectomy of the right true vocal cord were performed. Results The patient presented with a recurrence of the ΜFH alone (with no recurrence of the SCC) two months after the first operation and was managed with an extended second look laser cordectomy. The patient is under regular follow-up and remained disease-free nine months from diagnosis. Conclusions Our results show that early-stage simultaneous tumours of the larynx and particularly MFH and SCC can be treated efficiently with endoscopic laryngeal surgery alone. Close follow-up is of paramount importance because of the aggressive nature of MFH. PMID:27170836

  6. Titanium dioxide/silicon hole-blocking selective contact to enable double-heterojunction crystalline silicon-based solar cell

    SciTech Connect

    Nagamatsu, Ken A. Man, Gabriel; Jhaveri, Janam; Berg, Alexander H.; Kahn, Antoine; Wagner, Sigurd; Sturm, James C.; Avasthi, Sushobhan; Sahasrabudhe, Girija; Schwartz, Jeffrey

    2015-03-23

    In this work, we use an electron-selective titanium dioxide (TiO{sub 2}) heterojunction contact to silicon to block minority carrier holes in the silicon from recombining at the cathode contact of a silicon-based photovoltaic device. We present four pieces of evidence demonstrating the beneficial effect of adding the TiO{sub 2} hole-blocking layer: reduced dark current, increased open circuit voltage (V{sub OC}), increased quantum efficiency at longer wavelengths, and increased stored minority carrier charge under forward bias. The importance of a low rate of recombination of minority carriers at the Si/TiO{sub 2} interface for effective blocking of minority carriers is quantitatively described. The anode is made of a poly(3,4-ethylenedioxythiophene):poly(styrenesulfonate) heterojunction to silicon which forms a hole selective contact, so that the entire device is made at a maximum temperature of 100 °C, with no doping gradients or junctions in the silicon. A low rate of recombination of minority carriers at the Si/TiO{sub 2} interface is crucial for effective blocking of minority carriers. Such a pair of complementary carrier-selective heterojunctions offers a path towards high-efficiency silicon solar cells using relatively simple and near-room temperature fabrication techniques.

  7. Arabidopsis thalianaMRE11 is essential for activation of cell cycle arrest, transcriptional regulation and DNA repair upon the induction of double-stranded DNA breaks.

    PubMed

    Šamanić, I; Cvitanić, R; Simunić, J; Puizina, J

    2016-07-01

    Given the fundamental role of MRE11 in many aspects of DNA metabolism and signalling in eukaryotes, we analysed the impact of several MRE11 mutations on DNA damage response (DDR) and DNA repair in Arabidopsis thaliana. Three different atmre11 and an atatm-2 mutant lines, together with the wild type (WT), were compared using a new Arabidopsis genotoxic assay for in situ evaluation of genome integrity and DNA damage repair efficiency after double strand break (DSB) induction. The results showed that, despite the phenotypic differences and different lengths of the putative truncated AtMRE11 proteins, all three atmre11 and the atatm-2 mutant lines exhibited common hypersensitivity to bleomycin treatment, where they only slightly reduced mitotic activity, indicating a G2/M checkpoint abrogation. In contrast to the WT, which reduced the frequency of chromosomal aberrations throughout the recovery period after treatment, none of the three atmre11 and atatm-2 mutants recovered. Moreover, atmre11-3 mutants, similarly to atatm-2 mutants, failed to transcriptionally induce several DDR genes and had altered expression of the CYCB1;1::GUS protein. Nevertheless, numerous chromosomal fusions in the atmre11 mutants, observed after DNA damage induction, suggest intensive DNA repair activity. These results indicate that functional and full-length AtMRE11 is essential for activation of the cell cycle arrest, transcriptional regulation and DNA repair upon induction of DSB. PMID:27007017

  8. Can calcitonin nasal spray reduce the risk of recurrence of central giant cell granuloma of the jaws? A double-blind clinical trial.

    PubMed

    Tabrizi, R; Fardisi, S; Zamiri, B; Amanpour, S; Karagah, T

    2016-06-01

    Recurrence is a major problem following the treatment of aggressive central giant cell granuloma (CGCG). The aim of this study was to compare the frequency of recurrence between patients who received calcitonin nasal spray after curettage of CGCGs and those who did not. A double-blind clinical trial was designed. Patients were allocated to one of two groups: those in the calcitonin group underwent curettage and received calcitonin salmon nasal spray 200IU/day once a day for 3 months after surgery; those in the control group underwent curettage of CGCGs and received a placebo once a day for 3 months after surgery. All patients were followed for 5 years after surgery. Twenty-four patients were treated in the two groups. There was no difference in age, sex, tumour size, or tumour location between the two groups (P>0.05). Eight of the 24 patients (33.3%) had recurrences during the follow-up period: one in the calcitonin group (9.1%) and seven in the control group (53.8%). Analysis of the data demonstrated a significant difference between the two study groups (P=0.033). It appears that calcitonin nasal spray may reduce the frequency of recurrence in aggressive CGCGs in the mandible and maxilla. PMID:26993105

  9. Enhanced photoelectrical performance of dye-sensitized solar cells with double-layer TiO2 on perovskite SrTiO3 substrate

    NASA Astrophysics Data System (ADS)

    Liu, Qiuhong; Sun, Qiong; Zhang, Min; Li, Yang; Zhao, Mei; Dong, Lifeng

    2016-04-01

    In this research, perovskite SrTiO3 particles are synthesized by a hydrothermal method, and TiO2 with a double-layer structure is grown on the SrTiO3 surface by a hydrolysis-condensation process. Structural characterizations reveal that TiO2 comprises of two phases: anatase film at the bottom and single-crystal rutile nanorods grown along the [110] direction on top. The TiO2-SrTiO3 composite film is investigated as photoanode material for dye-sensitized solar cells. In comparison with pure TiO2 and SrTiO3, the composite photoanode shows a much better performance in photoelectric conversion efficiency (1.35 %), which is about 2 and 100 times as efficient as pure TiO2 and SrTiO3, respectively. This indicates that the composite structure can facilitate charge carrier transfer and reduce electron-hole recombination to enhance photoelectrical properties of TiO2-based photoanode materials.

  10. Electrical Conductivity of SrxVMoO6-y (x=0.0, 0.1, 0.2) Double Perovskite Solid Oxide Fuel Cell Anode

    NASA Astrophysics Data System (ADS)

    Childs, Nicholas; Weisenstein, Adam; Key, Camas; Sofie, Stephen; Smith, Richard

    2012-10-01

    Solid Oxide Fuel Cells (SOFCs) are suited for high efficiency power generation, fuel flexibility, high temperature electrolysis, closed loop regenerative systems, oxygen generation, and carbon dioxide reduction. These capabilities make the SOFC highly versatile for: primary/secondary power systems, advanced life support, and in-situ resource utilization which may all be desired for a forthcoming lunar return and Mars Exploration. A promising anode material for a SOFCs is the double perovskite Sr2-xVMoO6-y(x=0.0-0.2), due to its stability, electronic, and ionic conduction. Anodes of this material were prepared via a tape casting technique. Electrical conductivity was studied in reducing atmospheres at temperatures up to 800 ^oC. V and Mo valence states were indentified before and after annealing in a hydrogen environment. Samples exhibited metallic conduction with electrical conductivity of ˜10^4S/cm in a reducing atmosphere at 25 ^oC. A highly insulating SrMoO4 phase forms upon room temperature exposure to air. The SrMoO4 phase can be reduced above 400 ^oC resulting in an increase in conductivity.

  11. Homologous recombination in plant cells is enhanced by in vivo induction of double strand breaks into DNA by a site-specific endonuclease.

    PubMed Central

    Puchta, H; Dujon, B; Hohn, B

    1993-01-01

    Induction of double strand breaks (DSBs) is coupled to meiotic and mitotic recombination in yeast. We show that also in a higher eukaryote induction of DSBs is directly correlated with a strong enhancement of recombination frequencies. We cotransfected Nicotiana plumbaginifolia protoplasts with a plasmid carrying a synthetic I-SceI gene, coding for a highly sequence specific endonuclease, together with recombination substrates carrying an I-SceI-site adjacent to their homologous sequences. We measured efficiencies of extrachromosomal recombination, using a well established transient beta-glucuronidase (GUS) assay. GUS enzyme activities were strongly increased when a plasmid carrying the I-SceI gene in sense but not in antisense orientation with respect to the promoter was included in the transfections. The in vivo induced DSBs were detected in the recombination substrates by Southern blotting, demonstrating that the yeast enzyme is functional in plant cells. At high ratios of transfected I-SceI-genes to I-SceI-sites the majority of the I-SceI-sites in the recombination substrates are cleaved, indicating that the induction of the DSBs is the rate limiting step in the described recombination reaction. These results imply that in vivo induction of transient breaks at specific sites in the plant genome could allow foreign DNA to be targeted to these sites via homologous recombination. Images PMID:8255757

  12. Successful second transplantation with non-myeloablative conditioning using haploidentical donors for young patients after graft failure following double umbilical cord cell transplantation.

    PubMed

    Liu, Huilan; Wang, Xingbing; Geng, Liangquan; Tang, Baolin; Tong, Juan; Yao, Wen; Wang, Zuyi; Sun, Zimin

    2010-06-01

    GF is a common and life-threatening complication of UCBT. Here, we report that successful second transplantation of five patients using G-CSF-mobilized maternal stem cells with non-myeloablative conditioning after GF following double UCBT. The median interval between the two transplants were 38 days. The first transplantation was administered after myeloablative conditioning for hematologic malignancies (n=3), and rabbit ATG in combination with cyclophosphamide for SAA (n=2). The second conditioning consisted of Flu and ATG-based non-myeloablative regimen. All five patients acquired quick and sustained engraftment after the second transplant. Treatment-related toxicity was minimal. Three patients developed acute GVHD (>grade II=1). Three patients developed chronic GVHD (limited=1, extensive=2). Severe infectious episodes were significant but manageable. With a median follow-up of 713 days (592-1127), all patients have currently had an event-free survival. These results indicate that a second transplant with non-myeloablative conditioning using mother as the donor for young patient after GF is feasible. PMID:19732379

  13. Dose-intensified CHOP with rituximab (R-Double-CHOP) followed by consolidation high-dose chemotherapies for patients with advanced diffuse large B-cell lymphoma.

    PubMed

    Kurita, Daisuke; Miura, Katsuhiro; Nakagawa, Masaru; Ohtake, Shimon; Sakagami, Masashi; Uchino, Yoshihito; Takahashi, Hiromichi; Kiso, Satomi; Hojo, Atsuko; Kodaira, Hitomi; Yagi, Mai; Hirabayashi, Yukio; Kobayashi, Yujin; Iriyama, Noriyoshi; Kobayashi, Sumiko; Hatta, Yoshihiro; Kura, Yoshimasa; Sugitani, Masahiko; Takei, Masami

    2015-06-01

    Even after the advent of rituximab, clinical outcomes of conventional immuno-chemotherapy for high-risk diffuse large B-cell lymphoma (DLBCL) remain unsatisfactory. We retrospectively evaluated the efficacy and safety of R-Double-CHOP (R-D-CHOP), consisting of rituximab (375 mg/m(2), day -2), cyclophosphamide (750 mg/m(2), day 1, 2), doxorubicin (50 mg/m(2), day 1, 2), vincristine [1.4 mg/m(2) (maximum 2.0 mg/body), day 1], and prednisolone (50 mg/m(2), day 1-5), followed by consolidation high-dose chemotherapy. This treatment was given to 51 de novo DLBCL patients with a median age of 54 (range 19-65), who were categorized as high/high-intermediate risk by the age-adjusted International Prognostic Index. Treatment was given every 3 weeks up to three courses. The overall response and the complete response rate for R-D-CHOP were 94 and 78 %, respectively. A total of 30 responders proceeded to high-dose chemotherapy followed by autologous stem cell transplantation (HDC/ASCT), whereas 16 received high-dose methotrexate (HD-MTX) alternatively. The 3-year overall survival and the event-free survival for all patients were 78 and 61 %, respectively. Major adverse events included hematological toxicities, but there were no treatment-related deaths during the observation period. We conclude that the R-D-CHOP regimen followed by HDC/ASCT or HD-MTX is a promising treatment option for younger patients with highly advanced DLBCL. PMID:25776837

  14. Rescue of Munc18-1 and -2 Double Knockdown Reveals the Essential Functions of Interaction between Munc18 and Closed Syntaxin in PC12 Cells

    PubMed Central

    Han, Liping; Jiang, Tiandan; Han, Gayoung A.; Malintan, Nancy T.; Xie, Li; Wang, Li; Tse, Frederick W.; Gaisano, Herbert Y.; Collins, Brett M.; Meunier, Frederic A.

    2009-01-01

    Munc18-1 binds to syntaxin-1A via two distinct sites referred to as the “closed” conformation and N terminus binding. The latter has been shown to stimulate soluble N-ethylmaleimide-sensitive factor attachment protein receptor-mediated exocytosis, whereas the former is believed to be inhibitory or dispensable. To precisely define the contributions of each binding mode, we have engineered Munc18-1/-2 double knockdown neurosecretory cells and show that not only syntaxin-1A and -1B but also syntaxin-2 and -3 are significantly reduced as a result of Munc18-1 and -2 knockdown. Syntaxin-1 was mislocalized and the regulated secretion was abolished. We next examined the abilities of Munc18-1 mutants to rescue the defective phenotypes. Mutation (K46E/E59K) of Munc18-1 that selectively prevents binding to closed syntaxin-1 was unable to restore syntaxin-1 expression, localization, or secretion. In contrast, mutations (F115E/E132A) of Munc18-1 that selectively impair binding to the syntaxin-1 N terminus could still rescue the defective phenotypes. Our results indicate that Munc18-1 and -2 act in concert to support the expression of a broad range of syntaxins and to deliver syntaxin-1 to the plasma membrane. Our studies also indicate that the binding to the closed conformation of syntaxin is essential for Munc18-1 stimulatory action, whereas the binding to syntaxin N terminus plays a more limited role in neurosecretory cells. PMID:19812250

  15. Reduced DNA double-strand break repair capacity and risk of squamous cell carcinoma of the head and neck-A case-control study.

    PubMed

    Liu, Zhensheng; Liu, Hongliang; Gao, Fengqin; Dahlstrom, Kristina R; Sturgis, Erich M; Wei, Qingyi

    2016-04-01

    Tobacco smoke and alcohol use play important roles in the etiology of squamous cell carcinoma of the head and neck (SCCHN). Smoking causes DNA damage, including double-strand DNA breaks (DSBs), that leads to carcinogenesis. To test the hypothesis that suboptimal DSB repair capacity is associated with risk of SCCHN, we applied a flow cytometry-based method to detect the DSB repair phenotype first in four EBV-immortalized human lymphoblastoid cell lines and then in human peripheral blood T-lymphocytes (PBTLs). With this blood-based laboratory assay, we conducted a pilot case-control study of 100 patients with newly diagnosed, previously untreated SCCHN and 124 cancer-free controls of non-Hispanic whites. We found that the mean DSB repair capacity level was significantly lower in cases (42.1%) than that in controls (54.4%) (P<0.001). When we used the median DSB repair capacity level in the controls as the cutoff value for calculating the odds ratios (ORs) with adjustment for age, sex, smoking and drinking status, the cases were more likely than the controls to have a reduced DSB repair capacity (adjusted OR=1.93; 95% confidence interval, CI=1.04-3.56, P=0.037), especially for those subjects who were ever drinkers (adjusted OR=2.73; 95% CI=1.17-6.35, P=0.020) and had oropharyngeal tumors (adjusted OR=2.17; 95% CI=1.06-4.45, P=0.035). In conclusion, these findings suggest that individuals with a reduced DSB repair capacity may be at an increased risk of developing SCCHN. Larger studies are warranted to confirm these preliminary findings. PMID:26963119

  16. Unreacted PbI2 as a Double-Edged Sword for Enhancing the Performance of Perovskite Solar Cells.

    PubMed

    Jacobsson, T Jesper; Correa-Baena, Juan-Pablo; Halvani Anaraki, Elham; Philippe, Bertrand; Stranks, Samuel D; Bouduban, Marine E F; Tress, Wolfgang; Schenk, Kurt; Teuscher, Joël; Moser, Jacques-E; Rensmo, Håkan; Hagfeldt, Anders

    2016-08-17

    Lead halide perovskites have over the past few years attracted considerable interest as photo absorbers in PV applications with record efficiencies now reaching 22%. It has recently been found that not only the composition but also the precise stoichiometry is important for the device performance. Recent reports have, for example, demonstrated small amount of PbI2 in the perovskite films to be beneficial for the overall performance of both the standard perovskite, CH3NH3PbI3, as well as for the mixed perovskites (CH3NH3)x(CH(NH2)2)(1-x)PbBryI(3-y). In this work a broad range of characterization techniques including X-ray diffraction (XRD), scanning electron microscopy (SEM), transmission electron microscopy (TEM), photo electron spectroscopy (PES), transient absorption spectroscopy (TAS), UV-vis, electroluminescence (EL), photoluminescence (PL), and confocal PL mapping have been used to further understand the importance of remnant PbI2 in perovskite solar cells. Our best devices were over 18% efficient, and had in line with previous results a small amount of excess PbI2. For the PbI2-deficient samples, the photocurrent dropped, which could be attributed to accumulation of organic species at the grain boundaries, low charge carrier mobility, and decreased electron injection into the TiO2. The PbI2-deficient compositions did, however, also have advantages. The record Voc was as high as 1.20 V and was found in PbI2-deficient samples. This was correlated with high crystal quality, longer charge carrier lifetimes, and high PL yields and was rationalized as a consequence of the dynamics of the perovskite formation. We further found the ion migration to be obstructed in the PbI2-deficient samples, which decreased the JV hysteresis and increased the photostability. PbI2-deficient synthesis conditions can thus be used to deposit perovskites with excellent crystal quality but with the downside of grain boundaries enriched in organic species, which act as a barrier toward

  17. A double blind randomized placebo controlled phase I/II study assessing the safety and efficacy of allogeneic bone marrow derived mesenchymal stem cell in critical limb ischemia

    PubMed Central

    2013-01-01

    Background Peripheral vascular disease of the lower extremities comprises a clinical spectrum that extends from no symptoms to presentation with critical limb ischemia (CLI). Bone marrow derived Mesenchymal Stem Cells (BM- MSCs) may ameliorate the consequences of CLI due to their combinatorial potential for inducing angiogenesis and immunomodulatory environment in situ. The primary objective was to determine the safety of BM- MSCs in patients with CLI. Methods Prospective, double blind randomized placebo controlled multi-center study was conducted in patients with established CLI as per Rutherford classification in category II-4, III-5, or III-6 with infra-inguinal arterial occlusive disease and were not suitable for or had failed revascularization treatment. The primary end point was incidence of treatment – related adverse events (AE). Exploratory efficacy end points were improvement in rest pain, increase in Ankle Brachial Pressure Index (ABPI), ankle pressure, healing of ulcers, and amputation rates. Twenty patients (BM-MSC: Placebo = 1:1) were administered with allogeneic BM-MSCs at a dose of 2 million cells/kg or placebo (PlasmaLyte A) at the gastrocnemius muscle of the ischemic limb. Results Improvement was observed in the rest pain scores in both the arms. Significant increase in ABPI and ankle pressure was seen in BM-MSC arm compared to the placebo group. Incidence of AEs in the BM-MSC arm was 13 vs. 45 in the placebo arm where as serious adverse events (SAE) were similar in both the arms (5 in BM-MSC and 4 in the placebo group). SAEs resulted in death, infected gangrene, amputations in these patients. It was observed that the SAEs were related to disease progression and not related to stem cells. Conclusion BM-MSCs are safe when injected IM at a dose of 2 million cells/kg body weight. Few efficacy parameters such as ABPI and ankle pressure showed positive trend warranting further studies. Trial registration NIH website (http

  18. Damage to cellular DNA from particulate radiations, the efficacy of its processing and the radiosensitivity of mammalian cells. Emphasis on DNA double strand breaks and chromatin breaks

    NASA Technical Reports Server (NTRS)

    Lett, J. T.

    1992-01-01

    For several years, it has been evident that cellular radiation biology is in a necessary period of consolidation and transition (Lett 1987, 1990; Lett et al. 1986, 1987). Both changes are moving apace, and have been stimulated by studies with heavy charged particles. From the standpoint of radiation chemistry, there is now a consensus of opinion that the DNA hydration shell must be distinguished from bulk water in the cell nucleus and treated as an integral part of DNA (chromatin) (Lett 1987). Concomitantly, sentiment is strengthening for the abandonment of the classical notions of "direct" and "indirect" action (Fielden and O'Neill 1991; O'Neill 1991; O'Neill et al. 1991; Schulte-Frohlinde and Bothe 1991 and references therein). A layer of water molecules outside, or in the outer edge of, the DNA (chromatin) hydration shell influences cellular radiosensitivity in ways not fully understood. Charge and energy transfer processes facilitated by, or involving, DNA hydration must be considered in rigorous theories of radiation action on cells. The induction and processing of double stand breaks (DSBs) in DNA (chromatin) seem to be the predominant determinants of the radiotoxicity of normally radioresistant mammalian cells, the survival curves of which reflect the patterns of damage induced and the damage present after processing ceases, and can be modelled in formal terms by the use of reaction (enzyme) kinetics. Incongruities such as sublethal damage are neither scientifically sound nor relevant to cellular radiation biology (Calkins 1991; Lett 1990; Lett et al. 1987a). Increases in linear energy transfer (LET infinity) up to 100-200 keV micron-1 cause increases in the extents of neighboring chemical and physical damage in DNA denoted by the general term DSB. Those changes are accompanied by decreasing abilities of cells normally radioresistant to sparsely ionizing radiations to process DSBs in DNA and chromatin and to recover from radiation exposure, so they make

  19. A strategy of tumor treatment in mice with doxorubicin-cyclophosphamide combination based on dendritic cell activation by human double-stranded DNA preparation

    PubMed Central

    2010-01-01

    Background Immunization of mice with tumor homogenate after combined treatment with cyclophosphamide (CP) and double-stranded DNA (dsDNA) preparation is effective at inhibition of growth of tumor challenged after the treatment. It was assumed that this inhibition might be due to activation of the antigen-presenting cells. The purpose was to develop improved antitumor strategy using mice. We studied the combined action of cytostatics doxorubicin (Dox) plus CP with subsequent dsDNA preparation on tumor growth. Methods Three-month old CBA/Lac mice were used in the experiments. Mice were injected with CP and human dsDNA preparation. The percentage of mature dendritic cells (DCs) was estimated by staining of mononuclear cells isolated from spleen and bone marrow 3, 6, and 9 days later with monoclonal antibodies CD34, CD80, and CD86. In the next set of experiments, mice were given intramuscularly injections of 1-3 × 105 tumor cells. Four days later, they were injected intravenously with 6-6.7 mg/kg Dox and intraperitoneally with 100-200 mg/kg CP; 200 mkg human DNA was injected intraperitoneally after CP administration. Differences in tumor size between groups were analyzed for statistical significance by Student's t-test. The MTT-test was done to determine the cytotoxic index of mouse leucocytes from treated groups. Results The conducted experiments showed that combined treatment with CP and dsDNA preparation produce an increase in the total amount of mature DCs in vivo. Treatment of tumor bearers with preparation of fragmented dsDNA on the background of pretreatment with Dox plus CP demonstrated a strong suppression of tumor growth in two models. RLS, a weakly immunogenic, resistant to alkalyting cytostatics tumor, grew 3.4-fold slower when compared with the control (p < 0.001). In experiment with Krebs-2 tumor, only 2 of the 10 mice in the Dox+CP+DNA group had a palpable tumor on day 16. The cytotoxic index of leucocytes was 86.5% in the Dox+CP+DNA group, but it was 0

  20. Simulation of auroral double layers

    NASA Technical Reports Server (NTRS)

    Hubbard, R. F.; Joyce, G.

    1979-01-01

    Some basic properties of plasma double layers are deduced from a particle-in-cell computer simulation and related to parallel electric-field structures above the auroral regions. The simulation results on the processes leading to double-layer formation are examined, particularly in relation to the transient stage and double-layer structure and stability. It is concluded that: (1) a large potential difference applied to a finite-length plasma will be concentrated in a shocklike localized region instead of occurring over the entire length of the system; (2) the initial stage in double-layer formation is dominated by a large-potential pulse propagating in the direction of the induced electrostatic drift; (3) the entire potential is dropped over a specific scale length once the double layer has formed; and (4) this scale length is expected to be of the order of 1 km for a double layer above a discrete auroral arc with a potential of 10 kV and the electric-field vector parallel to the magnetic-field vector.

  1. Double bevel construction of a diamond anvil

    DOEpatents

    Moss, William C.

    1988-01-01

    A double or multiple bevel culet geometry is used on a diamond anvil in a high pressure cell apparatus to provide increased sample pressure and stability for a given force applied to the diamond tables. Double or multiple bevel culet geometries can also be used for sapphire or other hard crystal anvils. Pressures up to and above 5 Megabars can be reached.

  2. Double bevel construction of a diamond anvil

    DOEpatents

    Moss, W.C.

    1988-10-11

    A double or multiple bevel culet geometry is used on a diamond anvil in a high pressure cell apparatus to provide increased sample pressure and stability for a given force applied to the diamond tables. Double or multiple bevel culet geometries can also be used for sapphire or other hard crystal anvils. Pressures up to and above 5 Megabars can be reached. 8 figs.

  3. Lack of increased DNA double-strand breaks in peripheral blood mononuclear cells of individuals from high level natural radiation areas of Kerala coast in India.

    PubMed

    Jain, Vinay; Kumar, P R Vivek; Koya, P K M; Jaikrishan, G; Das, Birajalaxmi

    2016-06-01

    The high level natural radiation area (HLNRA) of Kerala is a 55km long and 0.5km wide strip in south west coast of India. The level of background radiation in this area varies from <1.0mGy/year to 45.0mGy/year. It offers unique opportunity to study the effect of chronic low dose/low dose-rate radiation directly on human population. Spontaneous level of DNA double strand breaks (DSBs) was quantified in peripheral blood mononuclear cells of 91 random individuals from HLNRA (N=61, mean age: 36.1±7.43years) and normal level natural radiation area (NLNRA) (N=30, mean age: 35.5±6.35years) using gamma-H2AX as a marker. The mean annual dose received by NLNRA and HLNRA individuals was 1.28±0.086mGy/year and 8.28±4.96mGy/year, respectively. The spontaneous frequency of DSBs in terms of gamma-H2AX foci among NLNRA and HLNRA individuals were 0.095±0.009 and 0.084±0.004 per cell (P=0.22). The individuals from HLNRA were further classified as low dose group (LDG, 1.51-5.0mGy/year, mean dose: 2.63±0.76mGy/year) and high dose group (HDG, >5.0mGy/year, mean dose: 11.04±3.57mGy/year). The spontaneous frequency of gamma-H2AX foci per cell in NLNRA, LDG and HDG was observed to be 0.095±0.009, 0.096±0.008 and 0.078±0.004 respectively. Individuals belonging to HDG of HLNRA showed marginally lower frequency of DSBs as compared to NLNRA and LDG of HLNRA. This could be suggestive of either lower induction or better repair of DSBs in individuals from HDG of HLNRA. The present study indicated that 5.0mGy/year could be a possible threshold dose for DSB induction at chronic low-dose radiation exposure in vivo. However, further studies on DNA damage induction and repair kinetics are required to draw firm conclusions. PMID:27063255

  4. Arg-Gly-Asp (RGD)-Modified E1A/E1B Double Mutant Adenovirus Enhances Antitumor Activity in Prostate Cancer Cells In Vitro and in Mice

    PubMed Central

    Wang, Hua; Cai, Zhi-Jian; Xu, Yi-Peng; Zhao, An; Su, Ying; Zhang, Gu; Zhu, Shao-Xing

    2016-01-01

    CAR is a transmembrane protein that is expressed in various epithelial and endothelial cells. CAR mediates adenoviral infection, as well as adenovirus-mediated oncolysis of AxdAdB-3, an E1A/E1B double-restricted oncolytic adenovirus, in prostate cancer cells. This study further assessed the therapeutic efficacy of AxdAdB-3 with Arg-Gly-Asp (RGD)-fiber modification (AxdAdB3-F/RGD), which enables integrin-dependent infection, in prostate cancer. Susceptibility of prostate cancer cells LNCaP, PC3, and DU145 to adenovirus infection was associated with CAR expression. All of the prostate cancer cell lines expressed integrin αvβ3 and αvβ5. AxdAdB-3 was more cytopathic in CAR-positive prostate cancer cells than in CAR-negative cells, whereas AxdAdB3-F/RGD caused potent oncolysis in both CAR-positive and CAR-negative prostate cancer cells. In contrast, AxdAdB3-F/RGD was not cytopathic against normal prostate epithelial cells, RWPE-1. Intratumoral injection of AxdAdB3-F/RGD into CAR-negative prostate cancer cell xenografts in nude mice inhibited tumor growth. The current study demonstrates that E1A/E1B double-restricted oncolytic adenovirus with an RGD-fiber modification enhances infection efficiency and anti-tumor activity in CAR-deficient prostate cancer cells, while sparing normal cells. Future studies will evaluate the therapeutic potential of AxdAdB3-F/RGD in prostate cancer. PMID:26799485

  5. Phosphorylation of murine double minute clone 2 (MDM2) protein at serine-267 by protein kinase CK2 in vitro and in cultured cells.

    PubMed

    Hjerrild, M; Milne, D; Dumaz, N; Hay, T; Issinger, O G; Meek, D

    2001-04-15

    Murine double minute clone 2 oncoprotein (MDM2) is a key component in the regulation of the tumour suppressor p53. MDM2 mediates the ubiqutination of p53 in the capacity of an E3 ligase and targets p53 for rapid degradation by the proteasome. Stress signals which impinge on p53, leading to its activation, promote disruption of the p53-MDM2 complex, as in the case of ionizing radiation, or block MDM2 synthesis and thereby reduce cellular MDM2 levels, as in the case of UV radiation. It is therefore likely that MDM2, which is known to be modified by ubiquitination, SUMOylation and multi-site phosphorylation, may itself be a target for stress signalling (SUMO is small ubiquitin-related modifier-1). In the present study we show that, like p53, the MDM2 protein is a substrate for phosphorylation by the protein kinase CK2 (CK2) in vitro. CK2 phosphorylates a single major site, Ser(267), which lies within the central acidic domain of MDM2. Fractionation of cellular extracts revealed the presence of a single Ser(267) protein kinase which co-purified with CK2 on ion-exchange chromatography and, like CK2, was subject to inhibition by micromolar concentrations of the CK2-specific inhibitor 5,6-dichlororibofuranosylbenzimidazole. Radiolabelling of cells expressing tagged recombinant wild-type MDM2 or a S267A (Ser(267)-->Ala) mutant, followed by phosphopeptide analysis, confirmed that Ser(267) is a cellular target for phosphorylation. Ser(267) mutants are still able to direct the degradation of p53, but in a slightly reduced capacity. These data highlight a potential route by which one of several physiological modifications occurring within the central acidic domain of the MDM2 protein can occur. PMID:11284721

  6. Reduction of Escherichia coli adherence to uroepithelial bladder cells after consumption of cranberry juice: a double-blind randomized placebo-controlled cross-over trial.

    PubMed

    Di Martino, P; Agniel, R; David, K; Templer, C; Gaillard, J L; Denys, P; Botto, H

    2006-02-01

    To determine the efficacy of the consumption of cranberry juice versus placebo with regard to the presence of in vitro bacterial anti-adherence activity in the urine of healthy volunteers. Twenty healthy volunteers, 10 men and 10 women, were included. The study was a double-blind, randomized, placebo-controlled, and cross-over study. In addition to normal diet, each volunteer received at dinner a single dose of 750 ml of a total drink composed of: (1) 250 ml of the placebo and 500 ml of mineral water, or (2) 750 ml of the placebo, or (3) 250 ml of the cranberry juice and 500 ml of mineral water, or (4) 750 ml of the cranberry juice. Each volunteer took the four regimens successively in a randomly order, with a washout period of at least 6 days between every change in regimen. The first urine of the morning following cranberry or placebo consumption was collected and used to support bacterial growth. Six uropathogenic Escherichia coli strains (all expressing type 1 pili; three positive for the gene marker for P-fimbriae papC and three negative for papC), previously isolated from patients with symptomatic urinary tract infections, were grown in urine samples and tested for their ability to adhere to the T24 bladder cell line in vitro. There were no significant differences in the pH or specific gravity between the urine samples collected after cranberry or placebo consumption. We observed a dose dependent significant decrease in bacterial adherence associated with cranberry consumption. Adherence inhibition was observed independently from the presence of genes encoding type P pili and antibiotic resistance phenotypes. Cranberry juice consumption provides significant anti-adherence activity against different E. coli uropathogenic strains in the urine compared with placebo. PMID:16397814

  7. Performance of (La,Sr)(Co,Fe)O 3- x double-layer cathode films for intermediate temperature solid oxide fuel cell

    NASA Astrophysics Data System (ADS)

    Marinha, D.; Hayd, J.; Dessemond, L.; Ivers-Tiffée, E.; Djurado, E.

    In this study the performance evaluation of (La,Sr)(Co,Fe)O 3- x (LSCF) double-layer films characterized by impedance spectroscopy between 403 and 603 °C to be used for intermediate temperature solid oxide fuel cells (IT-SOFCs) is presented. Two LSCF layers with different microstructures were sequentially deposited onto Ce 0.9Gd 0.1O 1.95 (CGO) substrates in a symmetrical fashion. A first layer of La 0.6Sr 0.4Co 0.2Fe 0.8O 3- x with a thickness of 7 μm and a nano-scaled particle size was deposited by electrostatic spray deposition (ESD) technique. Different deposition conditions were used in preparing the ESD films to evaluate the influence of film morphology on the electrochemical performance. After annealing, a current collector layer of La 0.58Sr 0.4Co 0.2Fe 0.8O 3- x with ∼45 μm in thickness and a larger particle size was deposited by screen printing. Area specific resistances (ASRs) were determined from impedance spectroscopy measurements performed in air between 403 and 603 °C, at 25 °C steps. A dependence of electrochemical performance on the morphology of the LSCF layer deposited by ESD was observed. The lowest ASR, measured during 130 h of isothermal dwelling at 603 °C, averaged 0.13 Ω cm 2 with negligible variation and is the lowest reported value for this composition, to the best of our knowledge. Reported results assure an excellent suitability of this type of assembly for IT-SOFCs.

  8. Real-time tracking of hydrogen peroxide secreted by live cells using MnO2 nanoparticles intercalated layered doubled hydroxide nanohybrids.

    PubMed

    Asif, Muhammad; Aziz, Ayesha; Dao, Anh Quang; Hakeem, Abdul; Wang, Haitao; Dong, Shuang; Zhang, Guoan; Xiao, Fei; Liu, Hongfang

    2015-10-22

    We report a facile and green method for the fabrication of new type of electrocatalysts based on MnO2 nanoparticles incorporated on MgAl LDH P-type semiconductive channel and explore its practical applications as high-performance electrode materials for electrochemical biosensor. A series of MgAl layered doubled hydroxide (LDH) nanohybrids with fixed Mg/Al (M(2+)/M(3+) atomic ratio of 3) and varied amount of MnCl2.4H2O are fabricated by a facile co-precipitation method. This approach demonstrates the combination of distinct properties including excellent intercalation features of LDH for entrapping nanoparticles and high loading of MnO2 nanoparticles in the host layers of LDH. Among all samples, Mn5-MgAl with 0.04% loaded manganese has a good crystalline morphology. A well-dispersed MnO2 nanoparticles encapsulated into the host matrix of hydrotalcite exhibit enhanced electrocatalytic activity towards the reduction of H2O2 as well as excellent stability, selectivity and reproducibility due to synergistic effect of good catalytic ability of MnO2 and conductive MgAl LDH. Glass carbon electrode (GCE) modified with Mn5-MgAl possesses a wide linear range of 0.05-78 mM, lowest detection limit 5 μM (S/N = 3) and detection sensitivity of 0.9352 μAmM(-1). This outstanding performance enables it to be used for real-time tracking of H2O2 secreted by live HeLa cells. This work may provide new insight in clinical diagnosis, on-site environmental analysis and point of care testing devices. PMID:26526908

  9. No improvement in suboptimal vitamin A status with a randomized, double-blind, placebo-controlled trial of vitamin A supplementation in children with sickle cell disease123

    PubMed Central

    Dougherty, Kelly A; Schall, Joan I; Kawchak, Deborah A; Green, Michael H; Ohene-Frempong, Kwaku; Zemel, Babette S; Stallings, Virginia A

    2012-01-01

    Background: Suboptimal vitamin A status is prevalent in children with type SS sickle cell disease (SCD-SS) and is associated with hospitalizations and poor growth and hematologic status. The supplemental vitamin A dose that optimizes suboptimal vitamin A status in this population is unknown. Objective: The efficacy of Recommended Dietary Allowance (RDA) doses (based on age and sex) of vitamin A (300, 400, or 600 μg retinyl palmitate/d) or vitamin A + zinc (10 or 20 mg zinc sulfate/d) compared with placebo to optimize vitamin A status was assessed in children aged 2.0–12.9 y with SCD-SS and a suboptimal baseline serum retinol concentration (<30 μg/dL). Design: In this randomized, double-blind, placebo-controlled trial, vitamin A status (serum retinol, prealbumin, retinol-binding protein, and relative-dose-response test) and disease-related illness events were assessed. Results: Twelve months of vitamin A supplementation at the doses recommended for healthy US children (based on age and sex) failed to improve serum retinol values in either group (vitamin A: n = 23; vitamin A + zinc: n = 18) compared with placebo (n = 21). By 12 mo, the increase (±SD) in serum retinol (3.6 ± 2.8 μg/dL) in those taking 600 μg vitamin A/d was significantly different from the decrease (±SD; −2.8 ± 2.4 μg/dL) in those taking 300 μg/d, which possibly suggests a dose-response relation (P < 0.05) with RDA doses. Conclusions: Compared with placebo, 12 mo of vitamin A supplementation at the RDA for healthy children did not improve serum retinol values in children with SCD-SS, which possibly suggests that higher doses are needed. However, the existence of alternative conclusions emphasizes the need for future research. PMID:22952182

  10. Normal karyotype acute myeloid leukemia patients with CEBPA double mutation have a favorable prognosis but no survival benefit from allogeneic stem cell transplant.

    PubMed

    Ahn, Jae-Sook; Kim, Jae-Young; Kim, Hyeoung-Joon; Kim, Yeo-Kyeoung; Lee, Seung-Shin; Jung, Sung-Hoon; Yang, Deok-Hwan; Lee, Je-Jung; Kim, Nan Young; Choi, Seung Hyun; Minden, Mark D; Jung, Chul Won; Jang, Jun-Ho; Kim, Hee Je; Moon, Joon Ho; Sohn, Sang Kyun; Won, Jong-Ho; Kim, Sung-Hyun; Kim, Dennis Dong Hwan

    2016-01-01

    Normal karyotype acute myeloid leukemia (NK-AML) with CCAAT/enhancer binding protein α (CEBPA) mutations is known to have a more favorable prognosis. However, direct comparison of the clinical significance according to consolidation therapy has not been widely performed in patients with NK-AML. A total of 404 patients with NK-AML who received intensive induction chemotherapy were included in the present study. Diagnostic samples from the patients were evaluated for CEBPA mutations by direct sequencing. CEBPA single (sm) or double mutation (dm) was observed in 27 (6.7 %) and 51 (12.6 %) patients, respectively. CEBPA (dm) was associated with GATA2 (mut), and it was less frequently associated with FLT3-ITD(pos), NPM1 (mut), and DNMT3A (mut) in comparison with CEBPA (wild) or CEBPA (sm) (all p values <0.05). On multivariate analysis, CEBPA (dm) (p = 0.007, OR 39.593) was an independent risk factor for achievement of complete remission (CR). With a median follow-up of 40.1 months, CEBPA (dm) showed a favorable overall survival (OS), event-free survival (EFS), and lower relapse incidence (RI) in comparison with CEBPA (wild) (all p values <0.005). Comparison of clinical outcome analyses (consolidation chemotherapy vs. allogeneic hematopoietic cell transplantation (HCT)) demonstrated the role of consolidation treatment in patients with CEBPA (dm). Allogeneic HCT was associated with lower EFS and RI and a trend of higher non-relapse mortality. However, there was no statistically significant difference in OS. In conclusion, CEBPA (dm) was associated with other molecular mutations. Consolidation chemotherapy alone may overcome higher relapse rates by reducing the treatment mortality and increasing survival after relapse events in patients with CEBPA (dm) in NK-AML. PMID:26537612

  11. Double patterning compliant logic design

    NASA Astrophysics Data System (ADS)

    Ma, Yuangsheng; Sweis, Jason; Bencher, Chris; Deng, Yunfei; Dai, Huixiong; Yoshida, Hidekazu; Gisuthan, Bimal; Kye, Jongwook; Levinson, Harry J.

    2011-04-01

    Double patterning technology (DPT) is the only solution to enable the scaling for advanced technology nodes before EUV or any other advanced patterning techniques become available. In general, there are two major double patterning techniques: one is Litho-Etch-Litho-Etch (LELE), and the other is sidewall spacer technology, a Self-Aligned Double Patterning technique (SADP). While numerous papers have previously demonstrated these techniques on wafer process capabilities and processing costs, more study needs to be done in the context of standard cell design flow to enable their applications in mass production. In this paper, we will present the impact of DPT on logic designs, and give a thorough discussion on how to make DPT-compliant constructs, placement and routing using examples with Cadence's Encounter Digital Implementation System (EDI System).

  12. Double-layer coating of SrCO3/TiO2 on nanoporous TiO2 for efficient dye-sensitized solar cells.

    PubMed

    Wang, Shutao; Zhang, Xi; Zhou, Gang; Wang, Zhong-Sheng

    2012-01-14

    Surface modification plays a crucial role in improving the efficiency of dye-sensitized solar cells (DSSCs), but the reported surface treatments are in general superior to the untreated TiO(2) but inferior to the typical TiCl(4)-treated TiO(2) in terms of solar cell performance. This work demonstrates a two-step treatment of the nanoporous titania surface with strontium acetate [Sr(OAc)(2)] and TiCl(4) in order, each step followed by sintering. An electronically insulating layer of SrCO(3) is formed on the TiO(2) surface via the Sr(OAc)(2) treatment and then a fresh TiO(2) layer is deposited on top of the SrCO(3) layer via the TiCl(4) treatment, corresponding to a double layer of Sr(OAc)(2)/TiO(2) coated on the TiO(2) surface. As compared to the typical TiCl(4)-treated DSSC, the Sr(OAc)(2)-TiCl(4) treated DSSC improves short-circuit photocurrent (J(sc)) by 17%, open-circuit photovoltage (V(oc)) by 2%, and power conversion efficiency by 20%. These results indicate that the Sr(OAc)(2)-TiCl(4) treatment is better than the often used TiCl(4) treatment for fabrication of efficient DSSCs. Charge density at open circuit and controlled intensity modulated photocurrent/photovoltage spectroscopy reveal that the two electrodes show almost same conduction band level but different electron diffusion coefficient and charge recombination rate constant. Owing to the blocking effect of the SrCO(3) layer on electron recombination with I(3)(-) ions, the charge recombination rate constant of the Sr(OAc)(2)-TiCl(4) treated DSSC is half that of the TiCl(4)-treated DSSC, accounting well for the difference of their V(oc). The improved J(sc) is also attributed to the middle SrCO(3) layer, which increases dye adsorption and may improve charge separation efficiency due to the blocking effect of SrCO(3) on charge recombination. PMID:22108906

  13. DNA double-strand breaks in mammalian cells exposed to gamma-rays and very heavy ions. Fragment-size distributions determined by pulsed-field gel electrophoresis.

    PubMed

    Kraxenberger, F; Weber, K J; Friedl, A A; Eckardt-Schupp, F; Flentje, M; Quicken, P; Kellerer, A M

    1998-07-01

    The spatial distribution of DNA double-strand breaks (DSB) was assessed after treatment of mammalian cells (V79) with densely ionizing radiation. Cells were exposed to beams of heavy charged particles (calcium ions: 6.9 MeV/u, 2.1.10(3) keV/microm; uranium ions: 9.0 MeV/u, 1.4.10(4) keV/microm) at the linear accelerator UNILAC of GSI, Darmstadt. DNA was isolated in agarose plugs and subjected to pulsed-field gel electrophoresis under conditions that separated DNA fragments of size 50 kbp to 5 Mbp. The measured fragment distributions were compared to those obtained after gamma-irradiation and were analyzed by means of a convolution and a deconvolution technique. In contrast to the finding for gamma-radiation, the distributions produced by heavy ions do not correspond to the random breakage model. Their marked overdispersion and the observed excess of short fragments reflect spatial clustering of DSB that extends over large regions of the DNA, up to several mega base pairs (Mbp). At fluences of 0.75 and 1.5/microm2, calcium ions produce nearly the same shape of fragment spectrum, merely with a difference in the amount of DNA entering the gel; this suggests that the DNA is fragmented by individual calcium ions. At a fluence of 0.8/microm2 uranium ions produce a profile that is shifted to smaller fragment sizes in comparison to the profile obtained at a fluence of 0.4/microm2; this suggests cumulative action of two separate ions in the formation of fragments. These observations are not consistent with the expectation that the uranium ions, with their much larger LET, should be more likely to produce single particle action than the calcium ions. However, a consideration of the greater lateral extension of the tracks of the faster uranium ions explains the observed differences; it suggests that the DNA is closely coiled so that even DNA locations several Mbp apart are usually not separated by less than 0. 1 or 0.2 microm. PMID:9728743

  14. A double-double/double-single computation package

    SciTech Connect

    Bailey, David H.

    2004-12-01

    The DDFUNIDSFUN software permits a new or existing Fortran-90 program to utilize double-double precision (approx. 31 digits) or double-single precision (approx. 14 digits) arithmetic. Double-double precision is required by a rapidly expandirtg body of scientific computations in physics and mathematics, for which the conventional 64-bit IEEE computer arithmetic (about 16 decimal digit accuracy) is not sufficient. Double-single precision permits users of systems that do not have hardware 64-bit IEEE arithmetic (such as some game systems) to perform arithmetic at a precision nearly as high as that of systems that do. Both packages run significantly faster Than using multiple precision or arbitrary precision software for this purpose. The package includes an extensive set of low-level routines to perform high-precision arithmetic, including routines to calculate various algebraic and transcendental functions, such as square roots, sin, ccc, exp, log and others. In addition, the package includes high-level translation facilities, so that Fortran programs can utilize these facilities by making only a few changes to conventional Fortran programs. In most cases, the only changes that are required are to change the type statements of variables that one wishes to be treated as multiple precision, plus a few other minor changes. The DDFUN package is similar in functionality to the double-double part of the GD package, which was previously written at LBNL. However, the DDFUN package is written exclusively in Fortran-90, thus avoidIng difficulties that some users experience when using GD, which includes both Fortran-90 and C++ code.

  15. A double-double/double-single computation package

    Energy Science and Technology Software Center (ESTSC)

    2004-12-01

    The DDFUNIDSFUN software permits a new or existing Fortran-90 program to utilize double-double precision (approx. 31 digits) or double-single precision (approx. 14 digits) arithmetic. Double-double precision is required by a rapidly expandirtg body of scientific computations in physics and mathematics, for which the conventional 64-bit IEEE computer arithmetic (about 16 decimal digit accuracy) is not sufficient. Double-single precision permits users of systems that do not have hardware 64-bit IEEE arithmetic (such as some game systems)more » to perform arithmetic at a precision nearly as high as that of systems that do. Both packages run significantly faster Than using multiple precision or arbitrary precision software for this purpose. The package includes an extensive set of low-level routines to perform high-precision arithmetic, including routines to calculate various algebraic and transcendental functions, such as square roots, sin, ccc, exp, log and others. In addition, the package includes high-level translation facilities, so that Fortran programs can utilize these facilities by making only a few changes to conventional Fortran programs. In most cases, the only changes that are required are to change the type statements of variables that one wishes to be treated as multiple precision, plus a few other minor changes. The DDFUN package is similar in functionality to the double-double part of the GD package, which was previously written at LBNL. However, the DDFUN package is written exclusively in Fortran-90, thus avoidIng difficulties that some users experience when using GD, which includes both Fortran-90 and C++ code.« less

  16. The morphology, proliferation rate, and population doubling time factor of adipose-derived mesenchymal stem cells cultured on to non-aqueous SiO2, TiO2, and hybrid sol-gel-derived oxide coatings.

    PubMed

    Marycz, Krzysztof; Krzak-Roś, Justyna; Donesz-Sikorska, Anna; Śmieszek, Agnieszka

    2014-11-01

    In recent years, much attention has been paid to the development of tissue engineering and regenerative medicine, especially when stem cells of various sources are concerned. In addition to the interest in mesenchymal stem cells isolated from bone marrow, recently more consideration has been given to stem cells isolated from adipose tissue (AdMSCs), due to their less invasive method of collection as well as their ease of isolation and culture. However, the development of regenerative medicine requires both the application of biocompatible material and the stem cells to accelerate the regeneration. In this study, we investigated the morphology, proliferation rate index (PRi), and population doubling time factor of adipose-derived mesenchymal stem cells cultured on non-aqueous sol-gel-derived SiO2, TiO2, and SiO2/TiO2 oxide coatings. The results indicated an increase in PRi of AdMSCs when cultured on to titanium dioxide, suggesting its high attractiveness for AdMSCs. In addition, the proper morphology and the shortest doubling time of AdMSCs were observed when cultured on titanium dioxide coating. PMID:24408867

  17. The Cambridge Double Star Atlas

    NASA Astrophysics Data System (ADS)

    MacEvoy, Bruce; Tirion, Wil

    2015-12-01

    Preface; What are double stars?; The binary orbit; Double star dynamics; Stellar mass and the binary life cycle; The double star population; Detecting double stars; Double star catalogs; Telescope optics; Preparing to observe; Helpful accessories; Viewing challenges; Next steps; Appendices: target list; Useful formulas; Double star orbits; Double star catalogs; The Greek alphabet.

  18. Compilation and evaluation of gas phase diffusion coefficients of reactive trace gases in the atmosphere: Volume 2. Diffusivities of organic compounds, pressure-normalised mean free paths, and average Knudsen numbers for gas uptake calculations

    NASA Astrophysics Data System (ADS)

    Tang, M. J.; Shiraiwa, M.; Poschl, U.; Cox, R. A.; Kalberer, M.

    2015-05-01

    Diffusion of organic vapours to the surface of aerosol or cloud particles is an important step for the formation and transformation of atmospheric particles. So far, however, a database of gas phase diffusion coefficients for organic compounds of atmospheric interest has not been available. In this work we have compiled and evaluated gas phase diffusivities (pressure-independent diffusion coefficients) of organic compounds reported by previous experimental studies, and we compare the measurement data to estimates obtained with Fuller's semi-empirical method. The difference between measured and estimated diffusivities are mostly < 10%. With regard to gas-particle interactions, different gas molecules, including both organic and inorganic compounds, exhibit similar Knudsen numbers (Kn) although their gas phase diffusivities may vary over a wide range. This is because different trace gas molecules have similar mean free paths in air at a given pressure. Thus, we introduce the pressure-normalised mean free path, λP ~ 100 nm atm, as a near-constant generic parameter that can be used for approximate calculation of Knudsen numbers as a simple function of gas pressure and particle diameter to characterise the influence of gas phase diffusion on the uptake of gases by aerosol or cloud particles. We use a kinetic multilayer model of gas-particle interaction to illustrate the effects of gas phase diffusion on the condensation of organic compounds with different volatilities. The results show that gas phase diffusion can play a major role in determining the growth of secondary organic aerosol particles by condensation of low-volatility organic vapours.

  19. Double outlet right ventricle

    MedlinePlus

    ... medlineplus.gov/ency/article/007328.htm Double outlet right ventricle To use the sharing features on this page, please enable JavaScript. Double outlet right ventricle (DORV) is a heart disease that is ...

  20. Chromosome doubling method

    DOEpatents

    Kato, Akio

    2006-11-14

    The invention provides methods for chromosome doubling in plants. The technique overcomes the low yields of doubled progeny associated with the use of prior techniques for doubling chromosomes in plants such as grasses. The technique can be used in large scale applications and has been demonstrated to be highly effective in maize. Following treatment in accordance with the invention, plants remain amenable to self fertilization, thereby allowing the efficient isolation of doubled progeny plants.

  1. Synthesis, Biological Evaluation, and Structure–Activity Relationships of Novel Substituted N-Phenyl Ureidobenzenesulfonate Derivatives Blocking Cell Cycle Progression in S-Phase and Inducing DNA Double-Strand Breaks

    PubMed Central

    2012-01-01

    Twenty-eight new substituted N-phenyl ureidobenzenesulfonate (PUB-SO) and 18 N-phenylureidobenzenesulfonamide (PUB-SA) derivatives were prepared. Several PUB-SOs exhibited antiproliferative activity at the micromolar level against the HT-29, M21, and MCF-7 cell lines and blocked cell cycle progression in S-phase similarly to cisplatin. In addition, PUB-SOs induced histone H2AX (γH2AX) phosphorylation, indicating that these molecules induce DNA double-strand breaks. In contrast, PUB-SAs were less active than PUB-SOs and did no