Sample records for haematoporphyrin derivative photobleaching

  1. Haematoporphyrin and OO'-diacetylhaematoporphyrin binding by serum and cellular proteins. Implications for the clearance of these photochemotherapeutic agents by cells.

    PubMed Central

    Smith, A; Neuschatz, T

    1983-01-01

    Haematoporphyrin derivative (HpD), a mixture of porphyrins, is currently used as a photochemotherapeutic agent in the treatment of neoplasias. The interaction of purified components of HpD with serum and cellular proteins was investigated using absorption and fluorescence spectroscopy. The interactions of haematoporphyrin and OO'-diacetylhaematoporphyrin with human albumin and with haemopexin, the two major serum porphyrin-binding proteins, show stoichiometries of 1 mol of porphyrin bound per mol of protein. The apparent dissociation constants, Kd, are in the range of 1-2 microM for albumin and 3-4 microM for haemopexin. These two major components of HpD would, after intravenous injection, bind to albumin and circulate in serum as albumin complexes. Free porphyrin rather than porphyrin bound to albumin interacts with Morris hepatoma tissue culture cells. A rapid high-affinity saturable transport system operates at free porphyrin concentrations of less than 2 microM. In addition, fluorescence spectra show that components in rat liver cytosol can bind haematoporphyrin and OO'-diacetylhaematoporphyrin and distinguish these binders from those present in rat serum. PMID:6225429

  2. Ca(2+)-mediated prostaglandin E2 induction reduces haematoporphyrin-derivative-induced cytotoxicity of T24 human bladder transitional carcinoma cells in vitro.

    PubMed Central

    Penning, L C; Keirse, M J; VanSteveninck, J; Dubbelman, T M

    1993-01-01

    The effects of haematoporphyrin-derivative-mediated photodynamic treatment on arachidonic acid metabolism and its relation to clonogenicity have been studied in human bladder-tumour cells. Photodynamic treatment resulted in a transient release of arachidonic acid-derived compounds; prostaglandin E2 (PGE2) and thromboxane B2 (TXB2) especially were strongly increased. This release was reduced by chelation of intracellular Ca2+ with Quin-2 or by lowering the extracellular Ca2+ concentration in the medium with EGTA, presumably resulting in inhibition of phospholipase A2. A similar reduction was obtained when indomethacin, an inhibitor of the cyclo-oxygenase pathway, was added prior to light exposure. These three treatments enhanced the photosensitivity, as revealed by the clonogenicity assay. Incubation with PGE2 prior to light exposure, but not with TXB2, protected against reproductive-cell death. The results of these experiments suggest that Ca(2+)-mediated activation of cyclo-oxygenase, resulting in increased levels of PGE2, participates in a cellular-defence mechanism against photodynamic cell killing. PMID:8503851

  3. Macroscopic singlet oxygen model incorporating photobleaching as an input parameter

    NASA Astrophysics Data System (ADS)

    Kim, Michele M.; Finlay, Jarod C.; Zhu, Timothy C.

    2015-03-01

    A macroscopic singlet oxygen model for photodynamic therapy (PDT) has been used extensively to calculate the reacted singlet oxygen concentration for various photosensitizers. The four photophysical parameters (ξ, σ, β, δ) and threshold singlet oxygen dose ([1O2]r,sh) can be found for various drugs and drug-light intervals using a fitting algorithm. The input parameters for this model include the fluence, photosensitizer concentration, optical properties, and necrosis radius. An additional input variable of photobleaching was implemented in this study to optimize the results. Photobleaching was measured by using the pre-PDT and post-PDT sensitizer concentrations. Using the RIF model of murine fibrosarcoma, mice were treated with a linear source with fluence rates from 12 - 150 mW/cm and total fluences from 24 - 135 J/cm. The two main drugs investigated were benzoporphyrin derivative monoacid ring A (BPD) and 2-[1-hexyloxyethyl]-2-devinyl pyropheophorbide-a (HPPH). Previously published photophysical parameters were fine-tuned and verified using photobleaching as the additional fitting parameter. Furthermore, photobleaching can be used as an indicator of the robustness of the model for the particular mouse experiment by comparing the experimental and model-calculated photobleaching ratio.

  4. Effects of Photobleaching on Microplastics

    NASA Astrophysics Data System (ADS)

    Ferrone, Salvatore; Sullivan, Kelley

    The presence of microplastics in our oceans and lakes is a contemporary environmental issue. A popular method for studying microplastics is fluorescence microscopy. We are studying the effects of fluorescence photo-bleaching on the imaging of microplastics. Our goal is to find out to what extent microplastics photo-bleach and if the photo-bleaching is recoverable. Photo-bleaching may entirely destroy the plastics' ability to fluoresce, hamper it for a short time, or have a minuscule effects. For this project, we consider the seven recyclable plastics. For each plastic type, we record a video of the micro-plastics for several hours under 405 nm light, then analyze and plot the image intensity as a function of time to see if the outputted light from the plastic decays over time. We then take single images at different time intervals to check if the intensity recovers. Our results will help set conditions under which fluorescence techniques can be used on microplastics. Undergraduate Student.

  5. Decreasing photobleaching by silver island films: application to muscle⋆

    PubMed Central

    Muthu, P.; Gryczynski, I.; Gryczynski, Z.; Talent, J.; Akopova, I.; Jain, K.; Borejdo, J.

    2007-01-01

    Recently it has become possible to study interactions between proteins at the level of single molecules. This requires collecting data from an extremely small volume, small enough to contain one molecule—typically of the order of attoliters (10−18 L). Collection of data from such a small volume with sufficiently high signal-to-noise ratio requires that the rate of photon detection per molecule be high. This calls for a large illuminating light flux, which in turn leads to rapid photobleaching of the fluorophores that are labeling the proteins. To decrease photobleaching, we measured fluorescence from a sample placed on coverslips coated with silver island films (SIF). SIF reduce photobleaching because they enhance fluorescence brightness and significantly decrease fluorescence lifetime. Increase in the brightness effectively decreases photobleaching because illumination can be attenuated to obtain the same fluorescence intensity. Decrease of lifetime decreases photobleaching because short lifetime minimizes the probability of oxygen attack while the fluorophore is in the excited state. The decrease of photobleaching was demonstrated in skeletal muscle. Myofibrils were labeled lightly with rhodamine–phalloidin, placed on coverslips coated with SIF, illuminated by total internal reflection, and observed through a confocal aperture. We show that SIF causes the intensity of phalloidin fluorescence to increase 4- to 5- fold and its fluorescence lifetime to decrease on average 23-fold. As a consequence, the rate of photobleaching of four or five molecules of actin of a myofibril on Olympus coverslips coated with SIF decreased at least 30-fold in comparison with photobleaching on an uncoated coverslip. Significant decrease of photobleaching makes the measurement of signal from a single cross-bridge of contracting muscle feasible. PMID:17531183

  6. Photobleaching of red fluorescence in oral biofilms.

    PubMed

    Hope, C K; de Josselin de Jong, E; Field, M R T; Valappil, S P; Higham, S M

    2011-04-01

    Many species of oral bacteria can be induced to fluoresce due to the presence of endogenous porphyrins, a phenomenon that can be utilized to visualize and quantify dental plaque in the laboratory or clinical setting. However, an inevitable consequence of fluorescence is photobleaching, and the effects of this on longitudinal, quantitative analysis of dental plaque have yet to be ascertained. Filter membrane biofilms were grown from salivary inocula or single species (Prevotella nigrescens and Prevotella intermedia). The mature biofilms were then examined in a custom-made lighting rig comprising 405 nm light-emitting diodes capable of delivering 220 W/m(2) at the sample, an appropriate filter and a digital camera; a set-up analogous to quantitative light-induced fluorescence digital. Longitudinal sets of images were captured and processed to assess the degradation in red fluorescence over time. Photobleaching was observed in all instances. The highest rates of photobleaching were observed immediately after initiation of illumination, specifically during the first minute. Relative rates of photobleaching during the first minute of exposure were 19.17, 13.72 and 3.43 arbitrary units/min for P. nigrescens biofilms, microcosm biofilm and P. intermedia biofilms, respectively. Photobleaching could be problematic when making quantitative measurements of porphyrin fluorescence in situ. Reducing both light levels and exposure time, in combination with increased camera sensitivity, should be the default approach when undertaking analyses by quantitative light-induced fluorescence digital. © 2010 John Wiley & Sons A/S.

  7. Photobleaching response of different sources of chromophoric dissolved organic matter exposed to natural solar radiation using absorption and excitation-emission matrix spectra.

    PubMed

    Zhang, Yunlin; Liu, Xiaohan; Osburn, Christopher L; Wang, Mingzhu; Qin, Boqiang; Zhou, Yongqiang

    2013-01-01

    CDOM biogeochemical cycle is driven by several physical and biological processes such as river input, biogeneration and photobleaching that act as primary sinks and sources of CDOM. Watershed-derived allochthonous (WDA) and phytoplankton-derived autochthonous (PDA) CDOM were exposed to 9 days of natural solar radiation to assess the photobleaching response of different CDOM sources, using absorption and fluorescence (excitation-emission matrix) spectroscopy. Our results showed a marked decrease in total dissolved nitrogen (TDN) concentration under natural sunlight exposure for both WDA and PDA CDOM, indicating photoproduction of ammonium from TDN. In contrast, photobleaching caused a marked increase in total dissolved phosphorus (TDP) concentration for both WDA and PDA CDOM. Thus TDN:TDP ratios decreased significantly both for WDA and PDA CDOM, which partially explained the seasonal dynamic of TDN:TDP ratio in Lake Taihu. Photobleaching rate of CDOM absorption a(254), was 0.032 m/MJ for WDA CDOM and 0.051 m/MJ for PDA CDOM from days 0-9, indicating that phototransformations were initially more rapid for the newly produced CDOM from phytoplankton than for the river CDOM. Extrapolation of these values to the field indicated that 3.9%-5.1% CDOM at the water surface was photobleached and mineralized every day in summer in Lake Taihu. Photobleaching caused the increase of spectral slope, spectral slope ratio and molecular size, indicating the CDOM mean molecular weight decrease which was favorable to further microbial degradation of mineralization. Three fluorescent components were validated in parallel factor analysis models calculated separately for WDA and PDA CDOM. Our study suggests that the humic-like fluorescence materials could be rapidly and easily photobleached for WDA and PDA CDOM, but the protein-like fluorescence materials was not photobleached and even increased from the transformation of the humic-like fluorescence substance to the protein

  8. Photobleaching Response of Different Sources of Chromophoric Dissolved Organic Matter Exposed to Natural Solar Radiation Using Absorption and Excitation–Emission Matrix Spectra

    PubMed Central

    Zhang, Yunlin; Liu, Xiaohan; Osburn, Christopher L.; Wang, Mingzhu; Qin, Boqiang; Zhou, Yongqiang

    2013-01-01

    CDOM biogeochemical cycle is driven by several physical and biological processes such as river input, biogeneration and photobleaching that act as primary sinks and sources of CDOM. Watershed-derived allochthonous (WDA) and phytoplankton-derived autochthonous (PDA) CDOM were exposed to 9 days of natural solar radiation to assess the photobleaching response of different CDOM sources, using absorption and fluorescence (excitation-emission matrix) spectroscopy. Our results showed a marked decrease in total dissolved nitrogen (TDN) concentration under natural sunlight exposure for both WDA and PDA CDOM, indicating photoproduction of ammonium from TDN. In contrast, photobleaching caused a marked increase in total dissolved phosphorus (TDP) concentration for both WDA and PDA CDOM. Thus TDN∶TDP ratios decreased significantly both for WDA and PDA CDOM, which partially explained the seasonal dynamic of TDN∶TDP ratio in Lake Taihu. Photobleaching rate of CDOM absorption a(254), was 0.032 m/MJ for WDA CDOM and 0.051 m/MJ for PDA CDOM from days 0–9, indicating that phototransformations were initially more rapid for the newly produced CDOM from phytoplankton than for the river CDOM. Extrapolation of these values to the field indicated that 3.9%–5.1% CDOM at the water surface was photobleached and mineralized every day in summer in Lake Taihu. Photobleaching caused the increase of spectral slope, spectral slope ratio and molecular size, indicating the CDOM mean molecular weight decrease which was favorable to further microbial degradation of mineralization. Three fluorescent components were validated in parallel factor analysis models calculated separately for WDA and PDA CDOM. Our study suggests that the humic-like fluorescence materials could be rapidly and easily photobleached for WDA and PDA CDOM, but the protein-like fluorescence materials was not photobleached and even increased from the transformation of the humic-like fluorescence substance to the protein

  9. SU-D-16A-07: Photobleaching Predicts Necrosis in Interstitial PDT

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

    Kim, M; Finlay, J; Liu, B

    Purpose: Dosimetry for PDT has proven to be a challenge thus far, and for prediction of PDT outcome, a singlet oxygen model based on fundamental photophysical parameters has been developed. Previously, the photobleaching effect of photosensitizers was taken into account in the singlet oxygen explicit dosimetry model; here we report of direct measurements of photobleaching in the same model to assess the conditions under which implicit dosimetry using photobleaching can serve as an intermediate surrogate for PDT damage. Methods: Fluorescence spectra were measured interstitially in sensitized mouse tumors prior to after irradiation via a cylindrical diffuser. Photobleaching was determined bymore » the relative decrease in fluorescence amplitude from the initial pre-treatment measurement. Spectra were analyzed by singular value decomposition to determine the photosensitizer concentration. Different photosensitizers were used to see the effect of photobleaching on PDT outcome and the impact of fluence on photobleaching. The drugs used were BPD (at two drug-light intervals), HPPH, and Photofrin. PDT outcome was determined by tumor necrosis radii measured upon sectioning and staining of treated tumors. Results: Post-PDT photosentizer concentrations were compared to initial pre-PDT photosensitizer concentrations, and the decrease was greater with a higher fluence measured during treatment. Furthermore, photobleaching and necrosis radius were found to be positively correlated. The relationship between photobleaching and necrosis radius is sensitizer-dependent, however the differences among sensitizers can be understood in terms of their respective photophysical parameters. Conclusions: Photobleaching is predictive of PDT outcome, but a comprehensive singlet oxygen model, has the potential to further improve the prediction of PDT outcome and the understanding of implicit dosimetry.« less

  10. Raman background photobleaching as a possible method of cancer diagnostics

    NASA Astrophysics Data System (ADS)

    Brandt, Nikolai N.; Brandt, Nikolai B.; Chikishev, Andrey Y.; Gangardt, Mihail G.; Karyakina, Nina F.

    2001-06-01

    Kinetics of photobleaching of background in Raman spectra of aqueous solutions of plant toxins ricin and ricin agglutinin, ricin binding subunit, and normal and malignant human blood serum were measured. For the excitation of the spectra cw and pulsed laser radiation were used. The spectra of Raman background change upon laser irradiation. Background intensity is lower for the samples with small molecular weight. The cyclization of amino acid residues in the toxin molecules as well as in human blood serum can be a reason of the Raman background. The model of the background photobleaching is proposed. The differences in photobleaching kinetics in the cases of cw and pulsed laser radiation are discussed. It is shown that Raman background photobleaching can be very informative for cancer diagnostics.

  11. A novel method to accurately locate and count large numbers of steps by photobleaching

    PubMed Central

    Tsekouras, Konstantinos; Custer, Thomas C.; Jashnsaz, Hossein; Walter, Nils G.; Pressé, Steve

    2016-01-01

    Photobleaching event counting is a single-molecule fluorescence technique that is increasingly being used to determine the stoichiometry of protein and RNA complexes composed of many subunits in vivo as well as in vitro. By tagging protein or RNA subunits with fluorophores, activating them, and subsequently observing as the fluorophores photobleach, one obtains information on the number of subunits in a complex. The noise properties in a photobleaching time trace depend on the number of active fluorescent subunits. Thus, as fluorophores stochastically photobleach, noise properties of the time trace change stochastically, and these varying noise properties have created a challenge in identifying photobleaching steps in a time trace. Although photobleaching steps are often detected by eye, this method only works for high individual fluorophore emission signal-to-noise ratios and small numbers of fluorophores. With filtering methods or currently available algorithms, it is possible to reliably identify photobleaching steps for up to 20–30 fluorophores and signal-to-noise ratios down to ∼1. Here we present a new Bayesian method of counting steps in photobleaching time traces that takes into account stochastic noise variation in addition to complications such as overlapping photobleaching events that may arise from fluorophore interactions, as well as on-off blinking. Our method is capable of detecting ≥50 photobleaching steps even for signal-to-noise ratios as low as 0.1, can find up to ≥500 steps for more favorable noise profiles, and is computationally inexpensive. PMID:27654946

  12. Photobleaching correction in fluorescence microscopy images

    NASA Astrophysics Data System (ADS)

    Vicente, Nathalie B.; Diaz Zamboni, Javier E.; Adur, Javier F.; Paravani, Enrique V.; Casco, Víctor H.

    2007-11-01

    Fluorophores are used to detect molecular expression by highly specific antigen-antibody reactions in fluorescence microscopy techniques. A portion of the fluorophore emits fluorescence when irradiated with electromagnetic waves of particular wavelengths, enabling its detection. Photobleaching irreversibly destroys fluorophores stimulated by radiation within the excitation spectrum, thus eliminating potentially useful information. Since this process may not be completely prevented, techniques have been developed to slow it down or to correct resulting alterations (mainly, the decrease in fluorescent signal). In the present work, the correction by photobleaching curve was studied using E-cadherin (a cell-cell adhesion molecule) expression in Bufo arenarum embryos. Significant improvements were observed when applying this simple, inexpensive and fast technique.

  13. A novel method to accurately locate and count large numbers of steps by photobleaching.

    PubMed

    Tsekouras, Konstantinos; Custer, Thomas C; Jashnsaz, Hossein; Walter, Nils G; Pressé, Steve

    2016-11-07

    Photobleaching event counting is a single-molecule fluorescence technique that is increasingly being used to determine the stoichiometry of protein and RNA complexes composed of many subunits in vivo as well as in vitro. By tagging protein or RNA subunits with fluorophores, activating them, and subsequently observing as the fluorophores photobleach, one obtains information on the number of subunits in a complex. The noise properties in a photobleaching time trace depend on the number of active fluorescent subunits. Thus, as fluorophores stochastically photobleach, noise properties of the time trace change stochastically, and these varying noise properties have created a challenge in identifying photobleaching steps in a time trace. Although photobleaching steps are often detected by eye, this method only works for high individual fluorophore emission signal-to-noise ratios and small numbers of fluorophores. With filtering methods or currently available algorithms, it is possible to reliably identify photobleaching steps for up to 20-30 fluorophores and signal-to-noise ratios down to ∼1. Here we present a new Bayesian method of counting steps in photobleaching time traces that takes into account stochastic noise variation in addition to complications such as overlapping photobleaching events that may arise from fluorophore interactions, as well as on-off blinking. Our method is capable of detecting ≥50 photobleaching steps even for signal-to-noise ratios as low as 0.1, can find up to ≥500 steps for more favorable noise profiles, and is computationally inexpensive. © 2016 Tsekouras et al. This article is distributed by The American Society for Cell Biology under license from the author(s). Two months after publication it is available to the public under an Attribution–Noncommercial–Share Alike 3.0 Unported Creative Commons License (http://creativecommons.org/licenses/by-nc-sa/3.0).

  14. Microfluidic flow cytometer for quantifying photobleaching of fluorescent proteins in cells

    PubMed Central

    Lubbeck, Jennifer L.; Dean, Kevin M.; Ma, Hairong; Palmer, Amy E.; Jimenez, Ralph

    2012-01-01

    Traditional flow cytometers are capable of rapid cellular assays on the basis of fluorescence intensity and light scatter. Microfluidic flow cytometers have largely followed the same path of technological development as their traditional counterparts, however the significantly smaller transport distance and resulting lower cell speeds in microchannels provides for the opportunity to detect novel spectroscopic signatures based on multiple, non-temporally-coincident excitation beams. Here, we characterize the design and operation of a cytometer with a 3-beam, probe/bleach/probe geometry, employing HeLa suspension cells expressing fluorescent proteins. The data collection rate exceeds 20 cells/s under a range of beam intensities (5 kW – 179 kW/cm2). The measured percent photobleaching (ratio of fluorescence intensities excited by the first and third beams: Sbeam3/Sbeam1) partially resolves a mixture of four red fluorescent proteins in mixed samples. Photokinetic simulations are presented and demonstrate that the percent photobleaching reflects a combination of the reversible and irreversible photobleaching kinetics. By introducing a photobleaching optical signature, which complements traditional fluorescence intensity-based detection, this method adds another dimension to multi-channel fluorescence cytometry, and provides a means for flow-cytometry-based screening of directed libraries of fluorescent protein photobleaching. PMID:22424298

  15. Background photobleaching in raman spectra of aqueous solutions of plant toxins

    NASA Astrophysics Data System (ADS)

    Brandt, Nikolai N.; Chikishev, Andrey Y.; Tonevitsky, Alexander G.

    2002-05-01

    Kinetics of background photobleaching in Raman spectra of aqueous solutions of ricin, ricin agglutinin and ricin binding subunit were measured. It was found that the spectrum of Raman background changes upon laser irradiation. Background intensity is lower for the samples with lower molecular weight. Photobleaching is characterized by oscillations in the multi exponentially decaying intensity.

  16. The fluorescent photobleaching properties of GFP expressed in human lung cancer cells

    NASA Astrophysics Data System (ADS)

    Jin, Ying; Xing, Da

    2003-12-01

    The characteristic properties of GFP make this protein a good candidate for use as a molecular reporter to monitor patterns of protein localization, gene expression, and intracellular protein trafficking in living cells. In this study, the dicistronic expression vector (pEGFP-C1) was used to transfected into human lung cancer cell line (ASTC-a-1) and a positive clone which stably expressed GFP in high level was obtained. After more than three months' passengers, the cells were also remained the strong fluorescence under fluorescent microscope. The results showed that the green fluorescent protein expressed in tumor cells was also photobleached under intense irradiation (approximately 488 nm) and the degree of photobleaching varied with the difference of the intensity of the excitation. Using different interdiction parcel (None, ND4, ND8, ND16), there were significant differences in photobleaching among the different excitation. The photobleaching was also affected by the time length of excitation, and the intensity of fluorescence was obviously decreased along with the increasing of excitation time, especially to stronger excitation.

  17. Resonant Scanning with Large Field of View Reduces Photobleaching and Enhances Fluorescence Yield in STED Microscopy.

    PubMed

    Wu, Yong; Wu, Xundong; Lu, Rong; Zhang, Jin; Toro, Ligia; Stefani, Enrico

    2015-10-01

    Photobleaching is a major limitation of superresolution Stimulated Depletion Emission (STED) microscopy. Fast scanning has long been considered an effective means to reduce photobleaching in fluorescence microscopy, but a careful quantitative study of this issue is missing. In this paper, we show that the photobleaching rate in STED microscopy can be slowed down and the fluorescence yield be enhanced by scanning with high speed, enabled by using large field of view in a custom-built resonant-scanning STED microscope. The effect of scanning speed on photobleaching and fluorescence yield is more remarkable at higher levels of depletion laser irradiance, and virtually disappears in conventional confocal microscopy. With ≥6 GW∙cm(-2) depletion irradiance, we were able to extend the fluorophore survival time of Atto 647N and Abberior STAR 635P by ~80% with 8-fold wider field of view. We confirm that STED Photobleaching is primarily caused by the depletion light acting upon the excited fluorophores. Experimental data agree with a theoretical model. Our results encourage further increasing the linear scanning speed for photobleaching reduction in STED microscopy.

  18. Excitation Light Dose Engineering to Reduce Photo-bleaching and Photo-toxicity

    PubMed Central

    Boudreau, Colton; Wee, Tse-Luen (Erika); Duh, Yan-Rung (Silvia); Couto, Melissa P.; Ardakani, Kimya H.; Brown, Claire M.

    2016-01-01

    It is important to determine the most effective method of delivering light onto a specimen for minimal light induced damage. Assays are presented to measure photo-bleaching of fluorophores and photo-toxicity to living cells under different illumination conditions. Turning the light off during part of the experimental time reduced photo-bleaching in a manner proportional to the time of light exposure. The rate of photo-bleaching of EGFP was reduced by 9-fold with light pulsing on the micro-second scale. Similarly, in living cells, rapid line scanning resulted in reduced cell stress as measured by mitochondrial potential, rapid cell protrusion and reduced cell retraction. This was achieved on a commercial confocal laser scanning microscope, without any compromise in image quality, by using rapid laser scan settings and line averaging. Therefore this technique can be implemented broadly without any software or hardware upgrades. Researchers can use the rapid line scanning option to immediately improve image quality on fixed samples, reduce photo-bleaching for large high resolution 3D datasets and improve cell health in live cell experiments. The assays developed here can be applied to other microscopy platforms to measure and optimize light delivery for minimal sample damage and photo-toxicity. PMID:27485088

  19. Excitation Light Dose Engineering to Reduce Photo-bleaching and Photo-toxicity.

    PubMed

    Boudreau, Colton; Wee, Tse-Luen Erika; Duh, Yan-Rung Silvia; Couto, Melissa P; Ardakani, Kimya H; Brown, Claire M

    2016-08-03

    It is important to determine the most effective method of delivering light onto a specimen for minimal light induced damage. Assays are presented to measure photo-bleaching of fluorophores and photo-toxicity to living cells under different illumination conditions. Turning the light off during part of the experimental time reduced photo-bleaching in a manner proportional to the time of light exposure. The rate of photo-bleaching of EGFP was reduced by 9-fold with light pulsing on the micro-second scale. Similarly, in living cells, rapid line scanning resulted in reduced cell stress as measured by mitochondrial potential, rapid cell protrusion and reduced cell retraction. This was achieved on a commercial confocal laser scanning microscope, without any compromise in image quality, by using rapid laser scan settings and line averaging. Therefore this technique can be implemented broadly without any software or hardware upgrades. Researchers can use the rapid line scanning option to immediately improve image quality on fixed samples, reduce photo-bleaching for large high resolution 3D datasets and improve cell health in live cell experiments. The assays developed here can be applied to other microscopy platforms to measure and optimize light delivery for minimal sample damage and photo-toxicity.

  20. Photoluminescence Intermittency and Photo-Bleaching of Single Colloidal Quantum Dot.

    PubMed

    Qin, Haiyan; Meng, Renyang; Wang, Na; Peng, Xiaogang

    2017-04-01

    Photoluminescence (PL) blinking of single colloidal quantum dot (QD)-PL intensity switching between different brightness states under constant excitation-and photo-bleaching are roadblocks for most applications of QDs. This progress report shall treat PL blinking and photo-bleaching both as photochemical events, namely, PL blinking as reversible and photo-bleaching being irreversible ones. Most studies on single-molecule spectroscopy of QDs in literature are related to PL blinking, which invites us to concentrate our discussions on the PL blinking, including its brief history in 20 years, analysis methods, competitive mechanisms and different strategies to battle it. In terms of suppression of the PL blinking, wavefunction confinement-confining photo-generated electron and hole within the core and inner portion of the shell of a core/shell QD-demonstrates significant advantages. This strategy yields nearly non-blinking QDs with their emission peaks covering most part of the visible window. As expected, the resulting QDs from this new strategy also show substantially improved anti-bleaching features. © 2017 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  1. Inferring subunit stoichiometry from single molecule photobleaching

    PubMed Central

    2013-01-01

    Single molecule photobleaching is a powerful tool for determining the stoichiometry of protein complexes. By attaching fluorophores to proteins of interest, the number of associated subunits in a complex can be deduced by imaging single molecules and counting fluorophore photobleaching steps. Because some bleaching steps might be unobserved, the ensemble of steps will be binomially distributed. In this work, it is shown that inferring the true composition of a complex from such data is nontrivial because binomially distributed observations present an ill-posed inference problem. That is, a unique and optimal estimate of the relevant parameters cannot be extracted from the observations. Because of this, a method has not been firmly established to quantify confidence when using this technique. This paper presents a general inference model for interpreting such data and provides methods for accurately estimating parameter confidence. The formalization and methods presented here provide a rigorous analytical basis for this pervasive experimental tool. PMID:23712552

  2. Staphyloxanthin photobleaching sensitizes methicillin-resistant Staphylococcus aureus to reactive oxygen species attack

    NASA Astrophysics Data System (ADS)

    Dong, Pu-Ting; Mohammad, Haroon; Hui, Jie; Wang, Xiaoyu; Li, Junjie; Liang, Lijia; Seleem, Mohamed N.; Cheng, Ji-Xin

    2018-02-01

    Given that the dearth of new antibiotic development loads an existential burden on successful infectious disease therapy, health organizations are calling for alternative approaches to combat methicillin-resistant Staphylococcus aureus (MRSA) infections. Here, we report a drug-free photonic approach to eliminate MRSA through photobleaching of staphyloxanthin, an indispensable membrane-bound antioxidant of S. aureus. The photobleaching process, uncovered through a transient absorption imaging study and quantitated by absorption spectroscopy and mass spectrometry, decomposes staphyloxanthin, and sensitizes MRSA to reactive oxygen species attack. Consequently, staphyloxanthin bleaching by low-level blue light eradicates MRSA synergistically with external or internal reactive oxygen species. The effectiveness of this synergistic therapy is validated in MRSA culture, MRSAinfected macrophage cells. Collectively, these findings highlight broad applications of staphyloxanthin photobleaching for treatment of MRSA infections.

  3. Raman Enhancement and Photo-Bleaching of Organic Dyes in the Presence of Chemical Vapor Deposition-Grown Graphene.

    PubMed

    Weng, Jiaxin; Zhao, Shichao; Li, Zhiting; Ricardo, Karen B; Zhou, Feng; Kim, Hyojeong; Liu, Haitao

    2017-10-19

    Fluorescent organic dyes photobleach under intense light. Graphene has been shown to improve the photo-stability of organic dyes. In this paper, we investigated the Raman spectroscopy and photo-bleaching kinetics of dyes in the absence/presence of chemical vapor deposition (CVD)-grown graphene. We show that graphene enhances the Raman signal of a wide range of dyes. The photo-bleaching of the dyes was reduced when the dyes were in contact with graphene. In contrast, monolayer hexagonal boron nitride (h-BN) was much less effective in reducing the photo-bleaching rate of the dyes. We attribute the suppression of photo-bleaching to the energy or electron transfer from dye to graphene. The results highlight the potential of CVD graphene as a substrate for protecting and enhancing Raman response of organic dyes.

  4. Visible light induced photobleaching of methylene blue over melamine-doped TiO2 nanocatalyst

    EPA Science Inventory

    TiO2 doping with N-rich melamine produced a stable, active and visible light sentisized nanocatalyst that showed a remarkable efficiency towards the photobleaching of a model compound – methylene blue (MB) in aqueous solution. The photobleaching followed a mixed reaction order ki...

  5. Axial superresolution via multiangle TIRF microscopy with sequential imaging and photobleaching

    PubMed Central

    Fu, Yan; Winter, Peter W.; Rojas, Raul; Wang, Victor; McAuliffe, Matthew; Patterson, George H.

    2016-01-01

    We report superresolution optical sectioning using a multiangle total internal reflection fluorescence (TIRF) microscope. TIRF images were constructed from several layers within a normal TIRF excitation zone by sequentially imaging and photobleaching the fluorescent molecules. The depth of the evanescent wave at different layers was altered by tuning the excitation light incident angle. The angle was tuned from the highest (the smallest TIRF depth) toward the critical angle (the largest TIRF depth) to preferentially photobleach fluorescence from the lower layers and allow straightforward observation of deeper structures without masking by the brighter signals closer to the coverglass. Reconstruction of the TIRF images enabled 3D imaging of biological samples with 20-nm axial resolution. Two-color imaging of epidermal growth factor (EGF) ligand and clathrin revealed the dynamics of EGF-activated clathrin-mediated endocytosis during internalization. Furthermore, Bayesian analysis of images collected during the photobleaching step of each plane enabled lateral superresolution (<100 nm) within each of the sections. PMID:27044072

  6. Depth-dependent autofluorescence photobleaching using 325, 473, 633, and 785 nm of porcine ear skin ex vivo

    NASA Astrophysics Data System (ADS)

    Schleusener, Johannes; Lademann, Jürgen; Darvin, Maxim E.

    2017-09-01

    Autofluorescence photobleaching describes the decrease of fluorescence intensity of endogenous fluorophores in biological tissue upon light irradiation. The origin of autofluorescence photobleaching is not fully understood. In the skin, the spatial distribution of various endogenous fluorophores varies within the skin layers. Most endogenous fluorophores are excited in the ultraviolet and short visible wavelength range, and only a few, such as porphyrins (red) and melanin (near-infrared), are excited at longer wavelengths. The excitation wavelength- and depth-dependent irradiation of skin will therefore excite different fluorophores, which will likely influence the photobleaching characteristics. The autofluorescence photobleaching of porcine ear skin has been measured ex vivo using 325, 473, 633, and 785 nm excitation at different skin depths from the surface to the dermis at 150 μm. Confocal Raman microscopes were used to achieve sufficient spatial resolution of the measurements. The autofluorescence area under the curve was measured for 21 consecutive acquisitions of 15 s. In all cases, the photobleaching follows a two-exponential decay function approximated by nonlinear regression. The results show that photobleaching can be applied to improve the signal-to-noise ratio in Raman spectroscopy for all of the applied excitation wavelengths and skin depths.

  7. Correction for photobleaching in dynamic fluorescence microscopy: application in the assessment of pharmacokinetic parameters in ultrasound-mediated drug delivery

    NASA Astrophysics Data System (ADS)

    Derieppe, M.; Bos, C.; de Greef, M.; Moonen, C.; de Senneville, B. Denis

    2016-01-01

    We have previously demonstrated the feasibility of monitoring ultrasound-mediated uptake of a hydrophilic model drug in real time with dynamic confocal fluorescence microscopy. In this study, we evaluate and correct the impact of photobleaching to improve the accuracy of pharmacokinetic parameter estimates. To model photobleaching of the fluorescent model drug SYTOX Green, a photobleaching process was added to the current two-compartment model describing cell uptake. After collection of the uptake profile, a second acquisition was performed when SYTOX Green was equilibrated, to evaluate the photobleaching rate experimentally. Photobleaching rates up to 5.0 10-3 s-1 were measured when applying power densities up to 0.2 W.cm-2. By applying the three-compartment model, the model drug uptake rate of 6.0 10-3 s-1 was measured independent of the applied laser power. The impact of photobleaching on uptake rate estimates measured by dynamic fluorescence microscopy was evaluated. Subsequent compensation improved the accuracy of pharmacokinetic parameter estimates in the cell population subjected to sonopermeabilization.

  8. Skin autofluorescence photo-bleaching and photo-memory

    NASA Astrophysics Data System (ADS)

    Lesins, Janis; Lihachev, Alexey; Rudys, Romualdas; Bagdonas, Saulius; Spigulis, Janis

    2011-07-01

    Photo-bleaching of in-vivo skin autofluorescence intensity under continuous low power laser irradiation has been studied. Temporal behavior of single-spot fluorescence and spectral fluorescent images have been studied at continuous 405 nm, 473 nm and 532 nm laser excitation and/or pre-irradiation, with power densities well below the laser-skin safety limits. Skin autofluorescence photo-memory effects (laser signatures) have been observed and analyzed, as well.

  9. Blueberry effects on dark vision and recovery after photobleaching: placebo-controlled crossover studies.

    PubMed

    Kalt, Wilhelmina; McDonald, Jane E; Fillmore, Sherry A E; Tremblay, Francois

    2014-11-19

    Clinical evidence for anthocyanin benefits in night vision is controversial. This paper presents two human trials investigating blueberry anthocyanin effects on dark adaptation, functional night vision, and vision recovery after retinal photobleaching. One trial, S2 (n = 72), employed a 3 week intervention and a 3 week washout, two anthocyanin doses (271 and 7.11 mg cyanidin 3-glucoside equivalents (C3g eq)), and placebo. The other trial, L1 (n = 59), employed a 12 week intervention and an 8 week washout and tested one dose (346 mg C3g eq) and placebo. In both S2 and L1 neither dark adaptation nor night vision was improved by anthocyanin intake. However, in both trials anthocyanin consumption hastened the recovery of visual acuity after photobleaching. In S2 both anthocyanin doses were effective (P = 0.014), and in L1 recovery was improved at 8 weeks (P = 0.027) and 12 weeks (P = 0.030). Although photobleaching recovery was hastened by anthocyanins, it is not known whether this improvement would have an impact on everyday vision.

  10. The photobleaching of the free and encapsulated metallic phthalocyanine and its effect on the photooxidation of simple molecules.

    PubMed

    Fanchiotti, Brenda Gomes; Machado, Marcella Piffer Zamprogno; de Paula, Letícia Camilato; Durmuş, Mahmut; Nyokong, Tebello; da Silva Gonçalves, Arlan; da Silva, André Romero

    2016-12-01

    The photobleaching of an unsubstituted phthalocyanine (gallium(III) phthalocyanine chloride (GaPc)) and a substituted phthalocyanine (1,4-(tetrakis[4-(benzyloxy)phenoxy]phthalocyaninato) indium(III) chloride (InTBPPc)) was monitored for the free photosensitizers and for the phthalocyanines encapsulated into nanoparticles of PEGylated poly(D,L-lactide-co-glycolide) (PLGA-PEG). Phosphate-buffered solutions (PBS) and organic solutions of the free GaPc or the free InTBPPc, and suspensions of each encapsulated photosensitizer (2-15μmol/L) were irradiated using a laser diode of 665nm with a power of 1-104mW and a light dose of 7.5J/cm 2 . The relative absorbance (RA) of the free GaPc dissolved in 1-methyl-2-pyrrolidone (MP) decreased 8.4 times when the laser power increased from 1mW to 104mW. However, the free or encapsulated GaPc did not suffer the photobleaching in PBS solution. The RA values decreased 2.4 times and 22.2 times for the free InTBPPc dissolved in PBS solution and in dimethylformamide (DMF), respectively, but the encapsulated InTBPPc was only photobleached when the laser power was 104mW at 8μmol/L. The increase of the free GaPc concentration favored the photobleaching in MP until 8μmol/L while the increase from 2μmol/L to 5μmol/L reduced the photodegradation in PBS solution. However, the photobleaching of the free InTBPPc in DMF or in PBS solution, and of each encapsulated photosensitizer was not influenced by increasing the concentration. The influence of the photobleaching on the capability of the free and encapsulated GaPc and InTBPPc to photooxidate the simple molecules was investigated monitoring the fluorescence of dimethylanthracene (DMA) and the tryptophan (Trp). Free InTBPPc was 2.0 and 1.8 times faster to photooxidate the DMA and Trp than it was the free GaPc, but the encapsulated GaPc was 3.4 times more efficient to photooxidize the Trp than it was the encapsulated InTBPPc due to the photodegradation suffered by the encapsulated In

  11. Combined electromagnetic and photoreaction modeling of CLD-1 photobleaching in polymer microring resonators

    NASA Astrophysics Data System (ADS)

    Huang, Yanyi; Poon, Joyce K. S.; Liang, Wei; Yariv, Amnon; Zhang, Cheng; Dalton, Larry R.

    2005-08-01

    By combining a solid-state photoreaction model with the modal solutions of an optical waveguide, we simulate the refractive index change due to the photobleaching of CLD-1 chromophores in an amorphous polycarbonate microring resonator. The simulation agrees well with experimental results. The photobleaching quantum efficiency of the CLD-1 chromophores is determined to be 0.65%. The combined modeling of the electromagnetic wave propagation and photoreaction precisely illustrates the spatial and temporal evolution of the optical properties of the polymer material as manifested in the refractive index and their effects on the modal and physical properties of the optical devices.

  12. Mobility measurement by analysis of fluorescence photobleaching recovery kinetics.

    PubMed Central

    Axelrod, D; Koppel, D E; Schlessinger, J; Elson, E; Webb, W W

    1976-01-01

    Fluorescence photobleaching recovery (FPR) denotes a method for measuring two-dimensional lateral mobility of fluorescent particles, for example, the motion of fluorescently labeled molecules in approximately 10 mum2 regions of a single cell surface. A small spot on the fluorescent surface is photobleached by a brief exposure to an intense focused laser beam, and the subsequent recovery of the fluorescence is monitored by the same, but attenuated, laser beam. Recovery occurs by replenishment of intact fluorophore in the bleached spot by lateral transport from the surrounding surface. We present the theoretical basis and some practical guidelines for simple, rigorous analysis of FPR experiments. Information obtainable from FPR experiments includes: (a) identification of transport process type, i.e. the admixture of random diffusion and uniform directed flow; (b) determination of the absolute mobility coefficient, i.e. the diffusion constant and/or flow velocity; and (c) the fraction of total fluorophore which is mobile. To illustrate the experimental method and to verify the theory for diffusion, we describe some model experiments on aqueous solutions of rhodamine 6G. PMID:786399

  13. Photobleaching dynamics in small molecule vs.  polymer organic photovoltaic blends with 1,7-bis-trifluoromethylfullerene

    DOE PAGES

    Garner, Logan E.; Nellissery Viswanathan, Vinila; Arias, Dylan H.; ...

    2018-02-27

    Two organic photovoltaic (OPV) donor materials (one polymer and one small molecule) are synthesized from the same constituent building blocks, namely thiophene units, cyclopentathiophene dione (CTD), and cyclopentadithiophene (CPDT). Photobleaching dynamics of these donor materials are then studied under white light illumination in air with blends of PC 70BM and the bistrifluoromethylfullerene 1,7-C 60(CF 3) 2. For both the polymer and small molecule blends, C 60(CF 3) 2 stabilizes the initial rate of photobleaching by a factor of 15 relative to PC70BM. However, once the small molecule:C 60(CF 3) 2 blend bleaches to ~80% of its initial optical density, themore » rate of photobleaching dramatically accelerates, which is not observed in the analagous polymer blend. We probe that phenomenon using time-resovled photoluminescence (TRPL) to measure PL quenching efficiencies at defined intervals during the photobleaching experiments. The data indicates the small molecule donor and C 60(CF 3) 2 acceptor significantly de-mix with time, after which the blend begins to bleach at approximately the same rate as the neat donor sample. The work suggests that perfluoroalkylfullerenes have great potential to stabilize certain OPV active layers toward photodegradation, provided their morphology is stable.« less

  14. Photobleaching dynamics in small molecule vs.  polymer organic photovoltaic blends with 1,7-bis-trifluoromethylfullerene

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

    Garner, Logan E.; Nellissery Viswanathan, Vinila; Arias, Dylan H.

    Two organic photovoltaic (OPV) donor materials (one polymer and one small molecule) are synthesized from the same constituent building blocks, namely thiophene units, cyclopentathiophene dione (CTD), and cyclopentadithiophene (CPDT). Photobleaching dynamics of these donor materials are then studied under white light illumination in air with blends of PC 70BM and the bistrifluoromethylfullerene 1,7-C 60(CF 3) 2. For both the polymer and small molecule blends, C 60(CF 3) 2 stabilizes the initial rate of photobleaching by a factor of 15 relative to PC70BM. However, once the small molecule:C 60(CF 3) 2 blend bleaches to ~80% of its initial optical density, themore » rate of photobleaching dramatically accelerates, which is not observed in the analagous polymer blend. We probe that phenomenon using time-resovled photoluminescence (TRPL) to measure PL quenching efficiencies at defined intervals during the photobleaching experiments. The data indicates the small molecule donor and C 60(CF 3) 2 acceptor significantly de-mix with time, after which the blend begins to bleach at approximately the same rate as the neat donor sample. The work suggests that perfluoroalkylfullerenes have great potential to stabilize certain OPV active layers toward photodegradation, provided their morphology is stable.« less

  15. Photobleaching of chromophoric dissolved organic matter (CDOM) in the Yangtze River estuary: kinetics and effects of temperature, pH, and salinity.

    PubMed

    Song, Guisheng; Li, Yijie; Hu, Suzheng; Li, Guiju; Zhao, Ruihua; Sun, Xin; Xie, Huixiang

    2017-06-21

    The kinetics and temperature-, pH- and salinity-dependences of photobleaching of chromophoric dissolved organic matter (CDOM) in the Yangtze River estuary (YRE) were evaluated using laboratory solar-simulated irradiation and compared to those of Suwannee River humic substances (SRHSs). Nearly all CDOM in water at the head of the estuary (headwater herein) was photobleachable in both summer and winter, while significant fractions of CDOM (13-29%) were resistant to photobleaching in saltier waters. The photobleaching rate constant in the headwater was 25% higher in summer than that in winter. The absorbed photon-based photobleaching efficiency (PE) increased with temperature following the linear Arrhenius equation. For a 20 °C increase in temperature, PE increased by ∼45% in the headwater and by 70-81% in the saltier waters. PE for YRE samples exhibited minima at pH from 6 to 7 and increased with both lower and higher pH values, contrasting the consistent increase in PE with pH shown by SRHSs. No consistent effect of salinity on PE was observed for both SRHSs and YRE samples. Photobleaching increased the spectral slope coefficient between 275 nm and 295 nm in summer, consistent with the behavior of SRHSs, but decreased it in winter, implying a difference in the molecular composition of chromophores between the two seasons. Temperature, salinity, and pH modified the photoalteration of the spectral shape but their effects varied spatially and seasonally. This study demonstrates that CDOM quality, temperature, and pH should be incorporated into models involving quantification of photobleaching.

  16. Power-law-distributed dark states are the main pathway for photobleaching of single organic molecules.

    PubMed

    Hoogenboom, Jacob P; van Dijk, Erik M H P; Hernando, Jordi; van Hulst, Niek F; García-Parajó, María F

    2005-08-26

    We exploit the strong excitonic coupling in a superradiant trimer molecule to distinguish between long-lived collective dark states and photobleaching events. The population and depopulation kinetics of the dark states in a single molecule follow power-law statistics over 5 orders of magnitude in time. This result is consistent with the formation of a radical unit via electron tunneling to a time-varying distribution of trapping sites in the surrounding polymer matrix. We furthermore demonstrate that this radicalization process forms the dominant pathway for molecular photobleaching.

  17. Analysis of Protein Kinetics Using Fluorescence Recovery After Photobleaching (FRAP).

    PubMed

    Giakoumakis, Nickolaos Nikiforos; Rapsomaniki, Maria Anna; Lygerou, Zoi

    2017-01-01

    Fluorescence recovery after photobleaching (FRAP) is a cutting-edge live-cell functional imaging technique that enables the exploration of protein dynamics in individual cells and thus permits the elucidation of protein mobility, function, and interactions at a single-cell level. During a typical FRAP experiment, fluorescent molecules in a defined region of interest within the cell are bleached by a short and powerful laser pulse, while the recovery of the fluorescence in the region is monitored over time by time-lapse microscopy. FRAP experimental setup and image acquisition involve a number of steps that need to be carefully executed to avoid technical artifacts. Equally important is the subsequent computational analysis of FRAP raw data, to derive quantitative information on protein diffusion and binding parameters. Here we present an integrated in vivo and in silico protocol for the analysis of protein kinetics using FRAP. We focus on the most commonly encountered challenges and technical or computational pitfalls and their troubleshooting so that valid and robust insight into protein dynamics within living cells is gained.

  18. Monitoring blood flow and photobleaching during topical ALA PDT treatment

    NASA Astrophysics Data System (ADS)

    Sands, Theresa L.; Sunar, Ulas; Foster, Thomas H.; Oseroff, Allan R.

    2009-02-01

    Photodynamic therapy (PDT) using topical aminolevulinic acid (ALA) is currently used as a clinical treatment for nonmelanoma skin cancers. In order to optimize PDT treatment, vascular shutdown early in treatment must be identified and prevented. This is especially important for topical ALA PDT where vascular shutdown is only temporary and is not a primary method of cell death. Shutdown in vasculature would limit the delivery of oxygen which is necessary for effective PDT treatment. Diffuse correlation spectroscopy (DCS) was used to monitor relative blood flow changes in Balb/C mice undergoing PDT at fluence rates of 10mW/cm2 and 75mW/cm2 for colon-26 tumors implanted intradermally. DCS is a preferable method to monitor the blood flow during PDT of lesions due to its ability to be used noninvasively throughout treatment, returning data from differing depths of tissue. Photobleaching of the photosensitizer was also monitored during treatment as an indirect manner of monitoring singlet oxygen production. In this paper, we show the conditions that cause vascular shutdown in our tumor model and its effects on the photobleaching rate.

  19. Segmented frequency-domain fluorescence lifetime measurements: minimizing the effects of photobleaching within a multi-component system.

    PubMed

    Marwani, Hadi M; Lowry, Mark; Keating, Patrick; Warner, Isiah M; Cook, Robert L

    2007-11-01

    This study introduces a newly developed frequency segmentation and recombination method for frequency-domain fluorescence lifetime measurements to address the effects of changing fractional contributions over time and minimize the effects of photobleaching within multi-component systems. Frequency segmentation and recombination experiments were evaluated using a two component system consisting of fluorescein and rhodamine B. Comparison of experimental data collected in traditional and segmented fashion with simulated data, generated using different changing fractional contributions, demonstrated the validity of the technique. Frequency segmentation and recombination was also applied to a more complex system consisting of pyrene with Suwannee River fulvic acid reference and was shown to improve recovered lifetimes and fractional intensity contributions. It was observed that photobleaching in both systems led to errors in recovered lifetimes which can complicate the interpretation of lifetime results. Results showed clear evidence that the frequency segmentation and recombination method reduced errors resulting from a changing fractional contribution in a multi-component system, and allowed photobleaching issues to be addressed by commercially available instrumentation.

  20. Kinetics of photobleaching of aqueous solutions of ricin agglutinin in the presence of guanidine chloride

    NASA Astrophysics Data System (ADS)

    Brandt, Nikolai N.; Chikishev, Andrey Y.

    2002-05-01

    Kinetics of background decay in Raman spectra of aqueous solutions of ricin agglutinin in the presence of guanidine chloride were measured. The differences in the kinetics of photobleaching are discussed.

  1. Analyzing Intracellular Binding and Diffusion with Continuous Fluorescence Photobleaching

    PubMed Central

    Wachsmuth, Malte; Weidemann, Thomas; Müller, Gabriele; Hoffmann-Rohrer, Urs W.; Knoch, Tobias A.; Waldeck, Waldemar; Langowski, Jörg

    2003-01-01

    Transport and binding of molecules to specific sites are necessary for the assembly and function of ordered supramolecular structures in cells. For analyzing these processes in vivo, we have developed a confocal fluorescence fluctuation microscope that allows both imaging of the spatial distribution of fluorescent molecules with confocal laser scanning microscopy and probing their mobility at specific positions in the cell with fluorescence correlation spectroscopy and continuous fluorescence photobleaching (CP). Because fluorescence correlation spectroscopy is restricted to rapidly diffusing particles and CP to slower processes, these two methods complement each other. For the analysis of binding-related contributions to mobility we have derived analytical expressions for the temporal behavior of CP curves from which the bound fraction and/or the dissociation rate or residence time at binding sites, respectively, can be obtained. In experiments, we investigated HeLa cells expressing different fluorescent proteins: Although enhanced green fluorescent protein (EGFP) shows high mobility, fusions of histone H2B with the yellow fluorescent protein are incorporated into chromatin, and these nuclei exhibit the presence of a stably bound and a freely diffusing species. Nonpermanent binding was found for mTTF-I, a transcription termination factor for RNA polymerase I, fused with EGFP. The cells show fluorescent nucleoli, and binding is transient. CP yields residence times for mTTF-I-EGFP of ∼13 s. PMID:12719264

  2. Analyzing intracellular binding and diffusion with continuous fluorescence photobleaching.

    PubMed

    Wachsmuth, Malte; Weidemann, Thomas; Müller, Gabriele; Hoffmann-Rohrer, Urs W; Knoch, Tobias A; Waldeck, Waldemar; Langowski, Jörg

    2003-05-01

    Transport and binding of molecules to specific sites are necessary for the assembly and function of ordered supramolecular structures in cells. For analyzing these processes in vivo, we have developed a confocal fluorescence fluctuation microscope that allows both imaging of the spatial distribution of fluorescent molecules with confocal laser scanning microscopy and probing their mobility at specific positions in the cell with fluorescence correlation spectroscopy and continuous fluorescence photobleaching (CP). Because fluorescence correlation spectroscopy is restricted to rapidly diffusing particles and CP to slower processes, these two methods complement each other. For the analysis of binding-related contributions to mobility we have derived analytical expressions for the temporal behavior of CP curves from which the bound fraction and/or the dissociation rate or residence time at binding sites, respectively, can be obtained. In experiments, we investigated HeLa cells expressing different fluorescent proteins: Although enhanced green fluorescent protein (EGFP) shows high mobility, fusions of histone H2B with the yellow fluorescent protein are incorporated into chromatin, and these nuclei exhibit the presence of a stably bound and a freely diffusing species. Nonpermanent binding was found for mTTF-I, a transcription termination factor for RNA polymerase I, fused with EGFP. The cells show fluorescent nucleoli, and binding is transient. CP yields residence times for mTTF-I-EGFP of approximately 13 s.

  3. Enzyme-dependent fluorescence recovery of NADH after photobleaching to assess dehydrogenase activity of isolated perfused hearts

    NASA Astrophysics Data System (ADS)

    Moreno, Angel; Kuzmiak-Glancy, Sarah; Jaimes, Rafael; Kay, Matthew W.

    2017-03-01

    Reduction of NAD+ by dehydrogenase enzymes to form NADH is a key component of cellular metabolism. In cellular preparations and isolated mitochondria suspensions, enzyme-dependent fluorescence recovery after photobleaching (ED-FRAP) of NADH has been shown to be an effective approach for measuring the rate of NADH production to assess dehydrogenase enzyme activity. Our objective was to demonstrate how dehydrogenase activity could be assessed within the myocardium of perfused hearts using NADH ED-FRAP. This was accomplished using a combination of high intensity UV pulses to photobleach epicardial NADH. Replenishment of epicardial NADH fluorescence was then imaged using low intensity UV illumination. NADH ED-FRAP parameters were optimized to deliver 23.8 mJ of photobleaching light energy at a pulse width of 6 msec and a duty cycle of 50%. These parameters provided repeatable measurements of NADH production rate during multiple metabolic perturbations, including changes in perfusate temperature, electromechanical uncoupling, and acute ischemia/reperfusion injury. NADH production rate was significantly higher in every perturbation where the energy demand was either higher or uncompromised. We also found that NADH production rate remained significantly impaired after 10 min of reperfusion after global ischemia. Overall, our results indicate that myocardial NADH ED-FRAP is a useful optical non-destructive approach for assessing dehydrogenase activity.

  4. Protoporphyrin IX Content Correlates with Activity of Photobleaching Herbicides

    PubMed Central

    Becerril, Jose M.; Duke, Stephen O.

    1989-01-01

    Several laboratories have demonstrated recently that photobleaching herbicides such as acifluorfen and oxadiazon cause accumulation of protoporphyrin IX (PPIX), a photodynamic pigment capable of herbicidal activity. We investigated, in acifluorfen-treated tissues, the in vivo stability of PPIX, the kinetics of accumulation, and the correlation between concentration of PPIX and herbicidal damage. During a 20 hour dark period, PPIX levels rose from barely detectable concentrations to 1 to 2 nanomoles per 50 cucumber (Cucumis sativus L.) cotyledon discs treated with 10 micromolar acifluorfen. When placed in 500 micromoles per square meter per second PAR, PPIX levels decayed logarithmically, with an initial half-life of about 2.5 hours. PPIX levels at each time after exposure to light correlated positively with the cellular damage that occurred during the following 1 hour in both green and yellow (tentoxin-treated) cucumber cotyledon tissues. PPIX levels in discs incubated for 20 hours in darkness correlated positively with the acifluorfen concentration in which they were incubated. In cucumber, the level of herbicidal damage caused by several p-nitrodiphenyl other herbicides, a p-chlorodiphenylether herbicide, and oxadiazon correlated positively with the amount of PPIX induced to accumulate by each of the herbicide treatments. Similar results were obtained with acifluorfen-treated pigweed and velvetleaf primary leaf tissues. In cucumber, PPIX levels increased within 15 and 30 minutes after exposure of discs to 10 micromolar acifluorfen in the dark and light, respectively. These data strengthen the view that PPIX is responsible for all or a major part of the photobleaching activity of acifluorfen and related herbicides. PMID:16666869

  5. Functional assessment of gap junctions in monolayer and three-dimensional cultures of human tendon cells using fluorescence recovery after photobleaching

    PubMed Central

    Kuzma-Kuzniarska, Maria; Yapp, Clarence; Pearson-Jones, Thomas W.; Jones, Andrew K.; Hulley, Philippa A.

    2014-01-01

    Abstract. Gap junction-mediated intercellular communication influences a variety of cellular activities. In tendons, gap junctions modulate collagen production, are involved in strain-induced cell death, and are involved in the response to mechanical stimulation. The aim of the present study was to investigate gap junction-mediated intercellular communication in healthy human tendon-derived cells using fluorescence recovery after photobleaching (FRAP). The FRAP is a noninvasive technique that allows quantitative measurement of gap junction function in living cells. It is based on diffusion-dependent redistribution of a gap junction-permeable fluorescent dye. Using FRAP, we showed that human tenocytes form functional gap junctions in monolayer and three-dimensional (3-D) collagen I culture. Fluorescently labeled tenocytes following photobleaching rapidly reacquired the fluorescent dye from neighboring cells, while HeLa cells, which do not communicate by gap junctions, remained bleached. Furthermore, both 18 β-glycyrrhetinic acid and carbenoxolone, standard inhibitors of gap junction activity, impaired fluorescence recovery in tendon cells. In both monolayer and 3-D cultures, intercellular communication in isolated cells was significantly decreased when compared with cells forming many cell-to-cell contacts. In this study, we used FRAP as a tool to quantify and experimentally manipulate the function of gap junctions in human tenocytes in both two-dimensional (2-D) and 3-D cultures. PMID:24390370

  6. Coumarin-Based Oxime Esters: Photobleachable and Versatile Unimolecular Initiators for Acrylate and Thiol-Based Click Photopolymerization under Visible Light-Emitting Diode Light Irradiation.

    PubMed

    Li, Zhiquan; Zou, Xiucheng; Zhu, Guigang; Liu, Xiaoya; Liu, Ren

    2018-05-09

    Developing efficient unimolecular visible light-emitting diode (LED) light photoinitiators (PIs) with photobleaching capability, which are essential for various biomedical applications and photopolymerization of thick materials, remains a great challenge. Herein, we demonstrate the synthesis of a series of novel PIs, containing coumarin moieties as chromophores and oxime ester groups as initiation functionalities and explore their structure-activity relationship. The investigated oxime esters can effectively induce acrylates and thiol-based click photopolymerization under 450 nm visible LED light irradiation. The initiator O-3 exhibited excellent photobleaching capability and enabled photopolymerization of thick materials (∼4.8 mm). The efficient unimolecular photobleachable initiators show great potential in dental materials and 3D printings.

  7. Fluorescence recovery after photo-bleaching as a method to determine local diffusion coefficient in the stratum corneum.

    PubMed

    Anissimov, Yuri G; Zhao, Xin; Roberts, Michael S; Zvyagin, Andrei V

    2012-10-01

    Fluorescence recovery after photo-bleaching experiments were performed in human stratum corneum in vitro. Fluorescence multiphoton tomography was used, which allowed the dimensions of the photobleached volume to be at the micron scale and located fully within the lipid phase of the stratum corneum. Analysis of the fluorescence recovery data with simplified mathematical models yielded the diffusion coefficient of small molecular weight organic fluorescent dye Rhodamine B in the stratum corneum lipid phase of about (3-6) × 10(-9)cm(2) s(-1). It was concluded that the presented method can be used for detailed analysis of localised diffusion coefficients in the stratum corneum phases for various fluorescent probes. Copyright © 2012 Elsevier B.V. All rights reserved.

  8. Measurement of the photobleaching kinetics of semiconducting polymer films by the pump - probe method

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

    Ozimova, A E; Bruevich, V V; Parashchuk, D Yu

    2011-12-31

    A phenomenological model of the laser photobleaching dynamics of a semiconducting polymer in a dual-beam scheme for different wavelengths of the burning and probe beams is developed. An experimental method is implemented based on this model, which allows one to investigate materials with significantly different photodegradation rates. The photodegradation quantum yield in mixtures of a semiconducting polymer belonging to polyparaphenylene vinylenes (MEH-PPV) with a low-molecular electron acceptor 2,4,7-trinitrofluorenone (TNF) is measured at burning wavelengths of 488 and 514 nm for different component ratios of MEHPPV : TNF. It is found that adding the acceptor decreases the polymer photodegradation quantum yieldmore » by at least four orders of magnitude in the MEH-PPV : TNF = 1 : 0.4 mixture; the photodegradation quantum yields are the same at both wavelengths. It is shown that the photodegradation rates of the MEH-PPV : TNF films measured by laser photobleaching and IR spectroscopy are in good agreement.« less

  9. Photocatalysis and self-catalyzed photobleaching with covalently-linked chromophore-quencher conjugates built around BOPHY.

    PubMed

    Sirbu, Dumitru; Woodford, Owen J; Benniston, Andrew C; Harriman, Anthony

    2018-06-13

    Two Chromophore-Quencher Conjugates (CQCs) have been synthesized by covalent attachment of the anti-oxidant dibutylated-hydroxytoluene (BHT) to a pyrrole-BF2 chromophore (BOPHY) in an effort to protect the latter against photofading. In fluid solution, light-induced intramolecular charge transfer is favoured in polar solvents and helps to inhibit photo-bleaching of the chromophore. The rate of photo-fading, which scales with the number of BHT residues, is zero-order in polar solvents but shows a linear dependence on the number of absorbed photons. The zero-order rate constant shows an inverse correlation with the fluorescence quantum yield measured in the same solvent. Photo-bleaching in benzonitrile involves autocatalysis while reaction in cyclohexane shows an unexpected stoichiometry. NMR spectroscopy indicates initial damage takes place at the BHT unit and allows identification of a reactive hydroperoxide as being the primary product. In the presence of an adventitious substrate, this hydroperoxide is a photocatalyst for amide formation under mild conditions.

  10. A Generalization of Theory for Two-Dimensional Fluorescence Recovery after Photobleaching Applicable to Confocal Laser Scanning Microscopes

    PubMed Central

    Kang, Minchul; Day, Charles A.; Drake, Kimberly; Kenworthy, Anne K.; DiBenedetto, Emmanuele

    2009-01-01

    Abstract Fluorescence recovery after photobleaching (FRAP) using confocal laser scanning microscopes (confocal FRAP) has become a valuable technique for studying the diffusion of biomolecules in cells. However, two-dimensional confocal FRAP sometimes yields results that vary with experimental setups, such as different bleaching protocols and bleaching spot sizes. In addition, when confocal FRAP is used to measure diffusion coefficients (D) for fast diffusing molecules, it often yields D-values that are one or two orders-of-magnitude smaller than that predicted theoretically or measured by alternative methods such as fluorescence correlation spectroscopy. Recently, it was demonstrated that this underestimation of D can be corrected by taking diffusion during photobleaching into consideration. However, there is currently no consensus on confocal FRAP theory, and no efforts have been made to unify theories on conventional and confocal FRAP. To this end, we generalized conventional FRAP theory to incorporate diffusion during photobleaching so that analysis by conventional FRAP theory for a circular region of interest is easily applicable to confocal FRAP. Finally, we demonstrate the accuracy of these new (to our knowledge) formulae by measuring D for soluble enhanced green fluorescent protein in aqueous glycerol solution and in the cytoplasm and nucleus of COS7 cells. PMID:19720039

  11. THE IMPACT OF CDOM PHOTOBLEACHING ON UV ATTENUATION NEAR CORAL REEFS IN THE FLORIDA KEYS

    EPA Science Inventory

    We have investigated how the loss of chromophoric dissolved organic matter (CDOM) in the water column due to photobleaching allows for increased penetration of UV radiation near coral reefs in the Florida Keys. Extended exposure to UV may contribute to coral bleaching episodes. C...

  12. Rapid and simple method of photobleaching to reduce background autofluorescence in lung tissue sections.

    PubMed

    Kumar, B Santhosh; Sandhyamani, S; Nazeer, Shaiju S; Jayasree, R S

    2015-02-01

    Autofluorescence exhibited by tissues often interferes with immunofluorescence. Using imaging and spectral analysis, we observed remarkable reduction of autofluorescence of formalin fixed paraffin embedded tissues irradiated with light prior to incubation with immunofluorescent dyes. The technique of photobleaching offers significant improvement in the quality and specificity of immunofluorescence. This has the potential for better techniques for disease diagnosis.

  13. Accelerated Photobleaching of a Cyanine Dye in the Presence of a Ternary Target DNA, PNA Probe, Dye Catalytic Complex: A Molecular Diagnostic

    PubMed Central

    Wang, M.; Holmes-Davis, R.; Rafinski, Z.; Jedrzejewska, B.; Choi, K. Y.; Zwick, M.; Bupp, C.; Izmailov, A.; Paczkowski, J.; Warner, B.; Koshinsky, H.

    2009-01-01

    In many settings, molecular testing is needed but unavailable due to complexity and cost. Simple, rapid, and specific DNA detection technologies would provide important alternatives to existing detection methods. Here we report a novel, rapid nucleic acid detection method based on the accelerated photobleaching of the light-sensitive cyanine dye, 3,3′-diethylthiacarbocyanine iodide (DiSC2(3) I−), in the presence of a target genomic DNA and a complementary peptide nucleic acid (PNA) probe. On the basis of the UV–vis, circular dichroism, and fluorescence spectra of DiSC2(3) with PNA–DNA oligomer duplexes and on characterization of a product of photolysis of DiSC2(3) I−, a possible reaction mechanism is proposed. We propose that (1) a novel complex forms between dye, PNA, and DNA, (2) this complex functions as a photosensitizer producing 1O2, and (3) the 1O2 produced promotes photobleaching of dye molecules in the mixture. Similar cyanine dyes (DiSC3(3), DiSC4(3), DiSC5(3), and DiSCpy(3)) interact with preformed PNA–DNA oligomer duplexes but do not demonstrate an equivalent accelerated photobleaching effect in the presence of PNA and target genomic DNA. The feasibility of developing molecular diagnostic assays based on the accelerated photobleaching (the smartDNA assay) that results from the novel complex formed between DiSC2(3) and PNA–DNA is under way. PMID:19231844

  14. Multiphoton versus confocal high resolution z-sectioning of enhanced green fluorescent microtubules: increased multiphoton photobleaching within the focal plane can be compensated using a Pockels cell and dual widefield detectors.

    PubMed

    Drummond, D R; Carter, N; Cross, R A

    2002-05-01

    Multiphoton excitation was originally projected to improve live cell fluorescence imaging by minimizing photobleaching effects outside the focal plane, yet reports suggest that photobleaching within the focal plane is actually worse than with one photon excitation. We confirm that when imaging enhanced green fluorescent protein, photobleaching is indeed more acute within the multiphoton excitation volume, so that whilst fluorescence increases as predicted with the square of the excitation power, photobleaching rates increase with a higher order relationship. Crucially however, multiphoton excitation also affords unique opportunities for substantial improvements to fluorescence detection. By using a Pockels cell to minimize exposure of the specimen together with multiple nondescanned detectors we show quantitatively that for any particular bleach rate multiphoton excitation produces significantly more signal than one photon excitation confocal microscopy in high resolution Z-axis sectioning of thin samples. Both modifications are readily implemented on a commercial multiphoton microscope system.

  15. Recent Applications of Fluorescence Recovery after Photobleaching (FRAP) to Membrane Bio-Macromolecules

    PubMed Central

    Rayan, Gamal; Guet, Jean-Erik; Taulier, Nicolas; Pincet, Frederic; Urbach, Wladimir

    2010-01-01

    This review examines some recent applications of fluorescence recovery after photobleaching (FRAP) to biopolymers, while mainly focusing on membrane protein studies. Initially, we discuss the lateral diffusion of membrane proteins, as measured by FRAP. Then, we talk about the use of FRAP to probe interactions between membrane proteins by obtaining fundamental information such as geometry and stoichiometry of the interacting complex. Afterwards, we discuss some applications of FRAP at the cellular level as well as the level of organisms. We conclude by comparing diffusion coefficients obtained by FRAP and several other alternative methods. PMID:22219695

  16. Characterization of Photofrin photobleaching for singlet oxygen dose estimation during photodynamic therapy of MLL cells in vitro

    NASA Astrophysics Data System (ADS)

    Dysart, Jonathan S.; Patterson, Michael S.

    2005-06-01

    A singlet oxygen dose model is developed for PDT with Photofrin. The model is based on photosensitizer photobleaching kinetics, and incorporates both singlet oxygen and non-singlet oxygen mediated bleaching mechanisms. To test our model, in vitro experiments were performed in which MatLyLu (MLL) cells were incubated in Photofrin and then irradiated with 532 nm light. Photofrin fluorescence was monitored during treatment and, at selected fluence levels, cell viability was determined using a colony formation assay. Cell survival correlated well to calculated singlet oxygen dose, independent of initial Photofrin concentration or oxygenation. About 2 × 108 molecules of singlet oxygen per cell were required to reduce the surviving fraction by 1/e. Analysis of the photobleaching kinetics suggests that the lifetime of singlet oxygen in cells is 0.048 ± 0.005 µs. The generation of fluorescent photoproducts was not a result of singlet oxygen reactions exclusively, and therefore did not yield additional information to aid in quantifying singlet oxygen dose.

  17. A Fluorescence Recovery After Photobleaching (FRAP) Technique for the Measurement of Solute Transport Across Surfactant-Laden Interfaces

    NASA Technical Reports Server (NTRS)

    Browne, Edward P.; Hatton, T. Alan

    1996-01-01

    The technique of Fluorescence Recovery After Photobleaching (FRAP) has been applied to the measurement of interfacial transport in two-phase systems. FRAP exploits the loss of fluorescence exhibited by certain fluorophores when over-stimulated (photobleached), so that a two-phase system, originally at equilibrium, can be perturbed without disturbing the interface by strong light from an argon-ion laser and its recovery monitored by a microscope-mounted CCD camera as it relaxes to a new equilibrium. During this relaxation, the concentration profiles of the probe solute are measured on both sides of the interface as a function of time, yielding information about the transport characteristics of the system. To minimize the size of the meniscus between the two phases, a photolithography technique is used to selectively treat the glass walls of the cell in which the phases are contained. This allows concentration measurements to be made very close to the interface and increases the sensitivity of the FRAP technique.

  18. Strategies to overcome photobleaching in algorithm-based adaptive optics for nonlinear in-vivo imaging.

    PubMed

    Caroline Müllenbroich, M; McGhee, Ewan J; Wright, Amanda J; Anderson, Kurt I; Mathieson, Keith

    2014-01-01

    We have developed a nonlinear adaptive optics microscope utilizing a deformable membrane mirror (DMM) and demonstrated its use in compensating for system- and sample-induced aberrations. The optimum shape of the DMM was determined with a random search algorithm optimizing on either two photon fluorescence or second harmonic signals as merit factors. We present here several strategies to overcome photobleaching issues associated with lengthy optimization routines by adapting the search algorithm and the experimental methodology. Optimizations were performed on extrinsic fluorescent dyes, fluorescent beads loaded into organotypic tissue cultures and the intrinsic second harmonic signal of these cultures. We validate the approach of using these preoptimized mirror shapes to compile a robust look-up table that can be applied for imaging over several days and through a variety of tissues. In this way, the photon exposure to the fluorescent cells under investigation is limited to imaging. Using our look-up table approach, we show signal intensity improvement factors ranging from 1.7 to 4.1 in organotypic tissue cultures and freshly excised mouse tissue. Imaging zebrafish in vivo, we demonstrate signal improvement by a factor of 2. This methodology is easily reproducible and could be applied to many photon starved experiments, for example fluorescent life time imaging, or when photobleaching is a concern.

  19. A modified GFP facilitates counting membrane protein subunits by step-wise photobleaching in Arabidopsis.

    PubMed

    Song, Kai; Xue, Yiqun; Wang, Xiaohua; Wan, Yinglang; Deng, Xin; Lin, Jinxing

    2017-06-01

    Membrane proteins exert functions by forming oligomers or molecular complexes. Currently, step-wise photobleaching has been applied to count the fluorescently labelled subunits in plant cells, for which an accurate and reliable control is required to distinguish individual subunits and define the basal fluorescence. However, the common procedure using immobilized GFP molecules is obviously not applicable for analysis in living plant cells. Using the spatial intensity distribution analysis (SpIDA), we found that the A206K mutation reduced the dimerization of GFP molecules. Further ectopic expression of Myristoyl-GFP A206K driven by the endogenous AtCLC2 promoter allowed imaging of individual molecules at a low expression level. As a result, the percentage of dimers in the transgenic pCLC2::Myristoyl-mGFP A206K line was significantly reduced in comparison to that of the pCLC2::Myristoyl-GFP line, confirming its application in defining the basal fluorescence intensity of GFP. Taken together, our results demonstrated that pCLC2::Myristoyl-mGFP A206K can be used as a standard control for monomer GFP, facilitating the analysis of the step-wise photobleaching of membrane proteins in Arabidopsis thaliana. Copyright © 2017 Elsevier GmbH. All rights reserved.

  20. Photoactive yellow protein from the purple phototrophic bacterium, Ectothiorhodospira halophila. Quantum yield of photobleaching and effects of temperature, alcohols, glycerol, and sucrose on kinetics of photobleaching and recovery.

    PubMed Central

    Meyer, T. E.; Tollin, G.; Hazzard, J. H.; Cusanovich, M. A.

    1989-01-01

    A water-soluble yellow protein from E. halophila was previously shown to be photoactive (Meyer, T. E., E. Yakali, M. A. Cusanovich, and G. Tollin. 1987. Biochemistry. 26:418-423). Pulsed laser excitation in the protein visible absorption band (maximum at 445 nm) causes a rapid bleach of color (k = 7.5 x 10(3) s-1) followed by a slower dark recovery (k = 2.6 s-1). This is analogous to the photocycle of sensory rhodopsin II from Halobacterium (which also has k = 2.6 s-1 for recovery). We have now determined the quantum yield of the photobleaching process to be 0.64, which is comparable with that of bacteriorhodopsin (0.25), and is thus large enough to be biologically significant. Although the photoreactions of yellow protein were previously shown to be relatively insensitive to pH, ionic strength and the osmoregulator betaine, the present experiments demonstrate that temperature, glycerol, sucrose, and various alcohol-water mixtures strongly influence the kinetics of photobleaching and recovery. The effect of temperature follows normal Arrhenius behavior for the bleach reaction (Ea = 15.5 kcal/mol). The rate constant for the recovery reaction increases with temperature between 5 degrees C and 35 degrees C, but decreases above 35 degrees C indicating alternate conformations with differing kinetics. There is an order of magnitude decrease in the rate constant for photobleaching in both glycerol and sucrose solutions that can be correlated with the changes in viscosity. We conclude from this that the protein undergoes a conformational change as a consequence of the photoinduced bleach. Recovery kinetics are affected by glycerol and sucrose to a much smaller extent and in a more complicated manner. Aliphatic, monofunctional alcohol-water solutions increase the rate constant for the bleach reaction and decrease the rate constant for the recovery reaction, each by an order of magnitude. These effects do not correlate with dielectric constant, indicating that the photocycle

  1. Measurement of resistance to solute transport across surfactant-laden interfaces using a Fluorescence Recovery After Photobleaching (FRAP) technique

    NASA Technical Reports Server (NTRS)

    Browne, Edward P.; Nivaggioli, Thierry; Hatton, T. Alan

    1994-01-01

    A noninvasive fluorescence recovery after photobleaching (FRAP) technique is under development to measure interfacial transport in two phase systems without disturbing the interface. The concentration profiles of a probe solute are measured in both sides of the interface by argon-ion laser, and the system relaxation is then monitored by a microscope-mounted CCD camera.

  2. Synthesis of photobleachable deep UV resists based on single component nonchemically amplified resist system

    NASA Astrophysics Data System (ADS)

    Kim, Kyoung-Seon; Kim, Su-Min; Park, Ji-Young; Kim, Jin-Baek

    2006-03-01

    In a general way, non-CARs consist of the matrix resins and photoactive compounds (PACs), and the dissolution properties of the resists are dependent on the amount of PACs. In common, I-line and G-line resists based on novolac and diazonaphthoquinone (DNQ) are typical non-CARs. But most PACs absorb much light in the deep UV, and they are poorly photobleached by deep UV exposure. This strong absorption of PACs prevents the deep UV light from reaching the bottom of the resist film, leading to scum and sloped pattern profiles. Several PACs which contain diazoketo groups have been reported for deep UV lithography. Our goal in this investigation is to find a proper resist that is processable without photoacid generator and induces both photobleaching in the deep UV regions and polarity change upon exposure. We thought diazoketo groups attached to the polymer side chains could give such effects. There is no necessity for the post-exposure bake step that is the cause of acid-diffusion. The diazoketo groups undergo the Wolff rearrangement upon irradiation in the deep UV, affording ketenes that react with water to provide base soluble photoproducts. The polymers were synthesized by radical copolymerization of 2-(2-diazo-3-oxo-butyryloxy)-ethyl methacrylate, 2-hydroxyethyl methacrylate, and γ-butyrolacton-2-yl methacrylate. The single component resist showed 0.7μm line and space patterns using a mercury-xenon lamp in a contact printing mode.

  3. Dynamic behavior of pump light radiation induced photo-bleaching effect on BAC-Si in bismuth/erbium co-doped optical fibers

    NASA Astrophysics Data System (ADS)

    Ding, Mingjie; Luo, Yanhua; Wen, Jianxiang; Peng, Gang-Ding

    2018-02-01

    Ultra-wide emission in bismuth doped optical fiber has been extremely studied for the development of the laser and amplifier working at near infrared band. In our homemade bismuth/erbium co-doped optical fiber, bismuth active center associated with silica (BAC-Si) has been found that when pumping at its resonant wavelength at 830 nm the NIR emission could be partially bleached. In addition, a self-recovery process has been observed at room temperature. However, the exact mechanism is still unclear. In this work, we have investigated the photo-bleaching effect on the BAC-Si via the pump power, pump wavelength and temperature dependence. Based on analyzing the result using stretched exponential function, it shows that the bleaching effect on BAC-Si has a strong link with the excitation process of Bi ion in BAC-Si. A potential energy curve model is used to illustrate the BAC-Si photo-bleaching process.

  4. Analytic solutions to modelling exponential and harmonic functions using Chebyshev polynomials: fitting frequency-domain lifetime images with photobleaching.

    PubMed

    Malachowski, George C; Clegg, Robert M; Redford, Glen I

    2007-12-01

    A novel approach is introduced for modelling linear dynamic systems composed of exponentials and harmonics. The method improves the speed of current numerical techniques up to 1000-fold for problems that have solutions of multiple exponentials plus harmonics and decaying components. Such signals are common in fluorescence microscopy experiments. Selective constraints of the parameters being fitted are allowed. This method, using discrete Chebyshev transforms, will correctly fit large volumes of data using a noniterative, single-pass routine that is fast enough to analyse images in real time. The method is applied to fluorescence lifetime imaging data in the frequency domain with varying degrees of photobleaching over the time of total data acquisition. The accuracy of the Chebyshev method is compared to a simple rapid discrete Fourier transform (equivalent to least-squares fitting) that does not take the photobleaching into account. The method can be extended to other linear systems composed of different functions. Simulations are performed and applications are described showing the utility of the method, in particular in the area of fluorescence microscopy.

  5. Wide-range tuning of polymer microring resonators by the photobleaching of CLD-1 chromophores

    NASA Astrophysics Data System (ADS)

    Poon, Joyce K. S.; Huang, Yanyi; Paloczi, George T.; Yariv, Amnon; Zhang, Cheng; Dalton, Larry R.

    2004-11-01

    We present a simple and effective method for the postfabrication trimming of optical microresonators. We photobleach CLD-1 chromophores to tune the resonance wavelengths of polymer microring resonator optical notch filters. A maximum wavelength shift of -8.73 nm is observed. The resonators are fabricated with a soft-lithography molding technique and have an intrinsic Q value of 2.6×10^4 and a finesse of 9.3. The maximum extinction ratio of the resonator filters is -34 dB, indicating that the critical coupling condition has been satisfied.

  6. Binding of the immunomodulatory drug Bz-423 to mitochondrial FoF1-ATP synthase in living cells by FRET acceptor photobleaching

    NASA Astrophysics Data System (ADS)

    Starke, Ilka; Johnson, Kathryn M.; Petersen, Jan; Gräber, Peter; Opipari, Anthony W.; Glick, Gary D.; Börsch, Michael

    2016-03-01

    Bz-423 is a promising new drug for treatment of autoimmune diseases. This small molecule binds to subunit OSCP of the mitochondrial enzyme FoF1-ATP synthase and modulates its catalytic activities. We investigate the binding of Bz-423 to mitochondria in living cells and how subunit rotation in FoF1-ATP synthase, i.e. the mechanochemical mechanism of this enzyme, is affected by Bz-423. Therefore, the enzyme was marked selectively by genetic fusion with the fluorescent protein EGFP to the C terminus of subunit γ. Imaging the threedimensional arrangement of mitochondria in living yeast cells was possible at superresolution using structured illumination microscopy, SIM. We measured uptake and binding of a Cy5-labeled Bz-423 derivative to mitochondrial FoF1-ATP synthase in living yeast cells using FRET acceptor photobleaching microscopy. Our data confirmed the binding of Cy5-labeled Bz-423 to the top of the F1 domain of the enzyme in mitochondria of living Saccharomyces cerevisiae cells.

  7. Validation of photodynamic action via photobleaching of a new curcumin-based composite with enhanced water solubility.

    PubMed

    Rego-Filho, Francisco G; de Araujo, Maria T; de Oliveira, Kleber T; Bagnato, Vanderlei S

    2014-09-01

    Motivated by the photochemical and photophysical properties of curcumin-based composites, the characteristics of a new curcumin-based water-soluble salt were investigated via absorption and fluorescence spectroscopy. Photobleaching was investigated using a set of LEDs in three different wavelengths (405 nm, 450 nm and 470 nm) to illuminate an aqueous solution of curcumin, evaluating its degradation for five different exposure times (0, 5, 15, 45 and 105 minutes). The results were compared with equivalent measurements of dark degradation and illumination in the presence of a singlet-oxygen quencher. Three solution concentrations (50, 100 and 150 μg/ml) were studied. To measure the fluorescence, it was used low power 405 nm excitation laser source. Time dependent photodegradation of curcumin was observed, as compared to the natural degradation of samples maintained on a dark environment. Two main absorption peaks were detected and their relation responded to both concentration and wavelength of the illumination source. A spectral correlation between absorption of curcumin and the emission bands of the sources showed an optimal spectral overlap for the 450 nm LED. For this source, photobleaching showed a less intense degradation on the presence of singlet oxygen quencher. This last result confirmed singlet oxygen production in vitro, indicating a strong potential of this composite to be used as a blue-light-activated photosensitizer.

  8. Analysis of protein and lipid dynamics using confocal fluorescence recovery after photobleaching (FRAP)

    PubMed Central

    Day, Charles A.; Kraft, Lewis J.; Kang, Minchul; Kenworthy, Anne K.

    2012-01-01

    Fluorescence recovery after photobleaching (FRAP) is a powerful, versatile and widely accessible tool to monitor molecular dynamics in living cells that can be performed using modern confocal microscopes. Although the basic principles of FRAP are simple, quantitative FRAP analysis requires careful experimental design, data collection and analysis. In this review we discuss the theoretical basis for confocal FRAP, followed by step-by-step protocols for FRAP data acquisition using a laser scanning confocal microscope for (1) measuring the diffusion of a membrane protein, (2) measuring the diffusion of a soluble protein, and (3) analysis of intracellular trafficking. Finally, data analysis procedures are discussed and an equation for determining the diffusion coefficient of a molecular species undergoing pure diffusion is presented. PMID:23042527

  9. EasyFRAP-web: a web-based tool for the analysis of fluorescence recovery after photobleaching data.

    PubMed

    Koulouras, Grigorios; Panagopoulos, Andreas; Rapsomaniki, Maria A; Giakoumakis, Nickolaos N; Taraviras, Stavros; Lygerou, Zoi

    2018-06-13

    Understanding protein dynamics is crucial in order to elucidate protein function and interactions. Advances in modern microscopy facilitate the exploration of the mobility of fluorescently tagged proteins within living cells. Fluorescence recovery after photobleaching (FRAP) is an increasingly popular functional live-cell imaging technique which enables the study of the dynamic properties of proteins at a single-cell level. As an increasing number of labs generate FRAP datasets, there is a need for fast, interactive and user-friendly applications that analyze the resulting data. Here we present easyFRAP-web, a web application that simplifies the qualitative and quantitative analysis of FRAP datasets. EasyFRAP-web permits quick analysis of FRAP datasets through an intuitive web interface with interconnected analysis steps (experimental data assessment, different types of normalization and estimation of curve-derived quantitative parameters). In addition, easyFRAP-web provides dynamic and interactive data visualization and data and figure export for further analysis after every step. We test easyFRAP-web by analyzing FRAP datasets capturing the mobility of the cell cycle regulator Cdt2 in the presence and absence of DNA damage in cultured cells. We show that easyFRAP-web yields results consistent with previous studies and highlights cell-to-cell heterogeneity in the estimated kinetic parameters. EasyFRAP-web is platform-independent and is freely accessible at: https://easyfrap.vmnet.upatras.gr/.

  10. Measuring Solvent Content of Macromolecular Crystals Using Fluorescence Recovery after Photobleaching

    NASA Astrophysics Data System (ADS)

    Siewny, Matthew; Kmetko, Jan

    2010-10-01

    We work out a novel protocol for measuring the solvent content (the fraction of crystal volume occupied by solvent) in biological crystals by the technique of fluorescence recovery after photobleaching (FRAP). Crystals of proteins with widely varying known solvent content (lysozyme, thaumatin, catalase, and ferritin) were grown in their native solution doped with sodium fluorescein dye and hydroxylamine (to prevent dye from binding to amine groups of the proteins.) The crystals were irradiated by a broadband, high intensity light through knife slits, leaving a rectangular area of bleached dye within the crystals. Measuring the flow of dye out of the bleached area allowed us to construct a curve relating the diffusion coefficient of dye to the channel size within the crystals, by solving the diffusion equation analytically. This curve may be used to measure the solvent content of any biological crystal in its native solution and help determine the number of proteins in the crystallographic asymmetric unit cell in x-ray structure solving procedures.

  11. Molecular counting by photobleaching in protein complexes with many subunits: best practices and application to the cellulose synthesis complex

    PubMed Central

    Chen, Yalei; Deffenbaugh, Nathan C.; Anderson, Charles T.; Hancock, William O.

    2014-01-01

    The constituents of large, multisubunit protein complexes dictate their functions in cells, but determining their precise molecular makeup in vivo is challenging. One example of such a complex is the cellulose synthesis complex (CSC), which in plants synthesizes cellulose, the most abundant biopolymer on Earth. In growing plant cells, CSCs exist in the plasma membrane as six-lobed rosettes that contain at least three different cellulose synthase (CESA) isoforms, but the number and stoichiometry of CESAs in each CSC are unknown. To begin to address this question, we performed quantitative photobleaching of GFP-tagged AtCESA3-containing particles in living Arabidopsis thaliana cells using variable-angle epifluorescence microscopy and developed a set of information-based step detection procedures to estimate the number of GFP molecules in each particle. The step detection algorithms account for changes in signal variance due to changing numbers of fluorophores, and the subsequent analysis avoids common problems associated with fitting multiple Gaussian functions to binned histogram data. The analysis indicates that at least 10 GFP-AtCESA3 molecules can exist in each particle. These procedures can be applied to photobleaching data for any protein complex with large numbers of fluorescently tagged subunits, providing a new analytical tool with which to probe complex composition and stoichiometry. PMID:25232006

  12. The Temporal Resolution of Laser Induced Fluorescence Photobleaching Anemometer

    NASA Astrophysics Data System (ADS)

    Zhao, Wei; Yang, Fang; Wang, Guiren

    2014-11-01

    Recently, in microfluidics, electrokinetic flows are widely used on micromixer designing. However, there is unfortunately no valid velocimeter today that can measure the random velocity fluctuation at high temporal and spatial resolution simultaneously in the complicated flow circumstance. We recently introduced laser induced fluorescence photobleaching anemometer (LIFPA), which has been successfully used in the measurement of velocity field in AC electrically driven microflow. Here, we theoretically study the temporal resolution (TR) of and experimentally verify, LIFPA can have simultaneously ultrahigh temporal (~4 μs) and spatial (~203 nm) resolution and can measure velocity fluctuation up to at least 2 kHz, whose corresponding wave number is about 6 × 106 1/m in an electrokinetically forced unsteady flow in microfluidics. The measurement of LIFPA is also compared with the widely used micro Particle Imaging Velocimetry (μPIV). We found, at the inlet, due to multiple uncertainties, the velocity fluctuations by μPIV exhibits apparently smaller values than that by LIFPA. But at downstreams, where velocity fluctuation is much lower than at the inlet and the uncertainties of complicated electric field on particles becomes smaller, LIFPA and μPIV indicate similar measurement. The work was supported by NSF under grant no. CAREER CBET-0954977 and MRI CBET-1040227, respectively.

  13. Modelling the competition between photo-darkening and photo-bleaching effects in high-power ytterbium-doped fibre amplifiers

    NASA Astrophysics Data System (ADS)

    Jolly, A.; Vinçont, C.; Pierre, Ch.; Boullet, J.

    2017-08-01

    We propose an innovative, fully space-time model to take into account the seed-dependent nature of ageing penalties in high-power ytterbium-doped fibre amplifiers. Ageing is shown to be based on the on-going competition between photo-darkening and photo-bleaching phenomena. Our approach is based on the natural interplay between the excited states of co-existing ytterbium pairs and colour centres in highly doped fibres, in the presence of thermal coupling between the closely spaced excited states. As initiated from IR photons, the excitation of colour centres up to the UV band is supposed to be governed by multi-photon absorption. The interactions of interest in the kinetics of photo-bleaching then take the form of highly efficient charge transfers, which imply the reduction of some fraction of the basically trivalent ions to their divalent state. Due to the activation of ytterbium pairs by means of energy transfer up-conversion, these interactions get more and more effective at elevated operating powers. Computational results using these principles actually help to fit our experimental data regarding seeding effects, as well as fully generic trends already evidenced in the literature. This gives a fine demonstration for the need to discriminate co-active pump and signal contributions. Our self-consistent, still simplified model then consists of a valuable tool to help for a deeper understanding of the ageing issues. Furthermore, considering higher-order ytterbium aggregates, this should open new routes towards more comprehensive models.

  14. Single molecule photobleaching (SMPB) technology for counting of RNA, DNA, protein and other molecules in nanoparticles and biological complexes by TIRF instrumentation.

    PubMed

    Zhang, Hui; Guo, Peixuan

    2014-05-15

    Direct counting of biomolecules within biological complexes or nanomachines is demanding. Single molecule counting using optical microscopy is challenging due to the diffraction limit. The single molecule photobleaching (SMPB) technology for direct counting developed by our team (Shu et al., 2007 [18]; Zhang et al., 2007 [19]) offers a simple and straightforward method to determine the stoichiometry of molecules or subunits within biocomplexes or nanomachines at nanometer scales. Stoichiometry is determined by real-time observation of the number of descending steps resulted from the photobleaching of individual fluorophore. This technology has now been used extensively for single molecule counting of protein, RNA, and other macromolecules in a variety of complexes or nanostructures. Here, we elucidate the SMPB technology, using the counting of RNA molecules within a bacteriophage phi29 DNA-packaging biomotor as an example. The method described here can be applied to the single molecule counting of other molecules in other systems. The construction of a concise, simple and economical single molecule total internal reflection fluorescence (TIRF) microscope combining prism-type and objective-type TIRF is described. The imaging system contains a deep-cooled sensitive EMCCD camera with single fluorophore detection sensitivity, a laser combiner for simultaneous dual-color excitation, and a Dual-View™ imager to split the multiple outcome signals to different detector channels based on their wavelengths. Methodology of the single molecule photobleaching assay used to elucidate the stoichiometry of RNA on phi29 DNA packaging motor and the mechanism of protein/RNA interaction are described. Different methods for single fluorophore labeling of RNA molecules are reviewed. The process of statistical modeling to reveal the true copy number of the biomolecules based on binomial distribution is also described. Copyright © 2014 Elsevier Inc. All rights reserved.

  15. Observation of photobleaching and intensity dependent kinetics in Ge22As22Se56 thin films under sub-bandgap light illumination

    NASA Astrophysics Data System (ADS)

    Khan, Pritam; Barik, A. R.; Vinod, E. M.; Sangunni, K. S.; Adarsh, K. V.

    2015-02-01

    We experimentally demonstrate photobleaching (PB) in Ge22As22Se56 thin films, when illuminated with a diode pumped solid state laser (DPSSL) of wavelength 671 nm, which is far below the optical bandgap of the sample. Interestingly, we found that PB is a slow process and occurs even at moderate pump beam intensity of 0.2 W/cm2, however the kinetics remain rather different.

  16. Molecular counting by photobleaching in protein complexes with many subunits: best practices and application to the cellulose synthesis complex

    DOE PAGES

    Chen, Yalei; Deffenbaugh, Nathan C.; Anderson, Charles T.; ...

    2014-09-17

    The constituents of large, multisubunit protein complexes dictate their functions in cells, but determining their precise molecular makeup in vivo is challenging. One example of such a complex is the cellulose synthesis complex (CSC), which in plants synthesizes cellulose, the most abundant biopolymer on Earth. In growing plant cells, CSCs exist in the plasma membrane as six-lobed rosettes that contain at least three different cellulose synthase (CESA) isoforms, but the number and stoichiometry of CESAs in each CSC are unknown. To begin to address this question, we performed quantitative photobleaching of GFP-tagged AtCESA3-containing particles in living Arabidopsis thaliana cells usingmore » variable-angle epifluorescence microscopy and developed a set of information-based step detection procedures to estimate the number of GFP molecules in each particle. The step detection algorithms account for changes in signal variance due to changing numbers of fluorophores, and the subsequent analysis avoids common problems associated with fitting multiple Gaussian functions to binned histogram data. The analysis indicates that at least 10 GFP-AtCESA3 molecules can exist in each particle. In conclusion, these procedures can be applied to photobleaching data for any protein complex with large numbers of fluorescently tagged subunits, providing a new analytical tool with which to probe complex composition and stoichiometry.« less

  17. Widefield Two-Photon Excitation without Scanning: Live Cell Microscopy with High Time Resolution and Low Photo-Bleaching

    PubMed Central

    Amor, Rumelo; McDonald, Alison; Trägårdh, Johanna; Robb, Gillian; Wilson, Louise; Abdul Rahman, Nor Zaihana; Dempster, John; Amos, William Bradshaw; Bushell, Trevor J.; McConnell, Gail

    2016-01-01

    We demonstrate fluorescence imaging by two-photon excitation without scanning in biological specimens as previously described by Hwang and co-workers, but with an increased field size and with framing rates of up to 100 Hz. During recordings of synaptically-driven Ca2+ events in primary rat hippocampal neurone cultures loaded with the fluorescent Ca2+ indicator Fluo-4 AM, we have observed greatly reduced photo-bleaching in comparison with single-photon excitation. This method, which requires no costly additions to the microscope, promises to be useful for work where high time-resolution is required. PMID:26824845

  18. Sequential photo-bleaching to delineate single Schwann cells at the neuromuscular junction.

    PubMed

    Brill, Monika S; Marinković, Petar; Misgeld, Thomas

    2013-01-11

    Sequential photo-bleaching provides a non-invasive way to label individual SCs at the NMJ. The NMJ is the largest synapse of the mammalian nervous system and has served as guiding model to study synaptic structure and function. In mouse NMJs motor axon terminals form pretzel-like contact sites with muscle fibers. The motor axon and its terminal are sheathed by SCs. Over the past decades, several transgenic mice have been generated to visualize motor neurons and SCs, for example Thy1-XFP and Plp-GFP mice, respectively. Along motor axons, myelinating axonal SCs are arranged in non-overlapping internodes, separated by nodes of Ranvier, to enable saltatory action potential propagation. In contrast, terminal SCs at the synapse are specialized glial cells, which monitor and promote neurotransmission, digest debris and guide regenerating axons. NMJs are tightly covered by up to half a dozen non-myelinating terminal SCs - these, however, cannot be individually resolved by light microscopy, as they are in direct membrane contact. Several approaches exist to individually visualize terminal SCs. None of these are flawless, though. For instance, dye filling, where single cells are impaled with a dye-filled microelectrode, requires destroying a labelled cell before filling a second one. This is not compatible with subsequent time-lapse recordings. Multi-spectral "Brainbow" labeling of SCs has been achieved by using combinatorial expression of fluorescent proteins. However, this technique requires combining several transgenes and is limited by the expression pattern of the promoters used. In the future, expression of "photo-switchable" proteins in SCs might be yet another alternative. Here we present sequential photo-bleaching, where single cells are bleached, and their image obtained by subtraction. We believe that this approach - due to its ease and versatility - represents a lasting addition to the neuroscientist's technology palette, especially as it can be used in vivo and

  19. Quantitative measurement of intracellular protein dynamics using photobleaching or photoactivation of fluorescent proteins.

    PubMed

    Matsuda, Tomoki; Nagai, Takeharu

    2014-12-01

    Unlike in vitro protein dynamics, intracellular protein dynamics are intricately regulated by protein-protein interactions or interactions between proteins and other cellular components, including nucleic acids, the plasma membrane and the cytoskeleton. Alteration of these dynamics plays a crucial role in physiological phenomena such as gene expression and cell division. Live-cell imaging via microscopy with the inherent properties of fluorescent proteins, i.e. photobleaching and photoconversion, or fluorescence correlation spectroscopy, provides insight into the movement of proteins and their interactions with cellular components. This article reviews techniques based on photo-induced changes in the physicochemical properties of fluorescent proteins to measure protein dynamics inside living cells, and it also discusses the strengths and weaknesses of these techniques. © The Author 2014. Published by Oxford University Press on behalf of The Japanese Society of Microscopy. All rights reserved. For permissions, please e-mail: journals.permissions@oup.com.

  20. Anomalous subdiffusion in fluorescence photobleaching recovery: a Monte Carlo study.

    PubMed Central

    Saxton, M J

    2001-01-01

    Anomalous subdiffusion is hindered diffusion in which the mean-square displacement of a diffusing particle is proportional to some power of time less than one. Anomalous subdiffusion has been observed for a variety of lipids and proteins in the plasma membranes of a variety of cells. Fluorescence photobleaching recovery experiments with anomalous subdiffusion are simulated to see how to analyze the data. It is useful to fit the recovery curve with both the usual recovery equation and the anomalous one, and to judge the goodness of fit on log-log plots. The simulations show that the simplest approximate treatment of anomalous subdiffusion usually gives good results. Three models of anomalous subdiffusion are considered: obstruction, fractional Brownian motion, and the continuous-time random walk. The models differ significantly in their behavior at short times and in their noise level. For obstructed diffusion the approach to the percolation threshold is marked by a large increase in noise, a broadening of the distribution of diffusion coefficients and anomalous subdiffusion exponents, and the expected abrupt decrease in the mobile fraction. The extreme fluctuations in the recovery curves at and near the percolation threshold result from extreme fluctuations in the geometry of the percolation cluster. PMID:11566793

  1. Fluorescence Recovery After Photobleaching Analysis of the Diffusional Mobility of Plasma Membrane Proteins: HER3 Mobility in Breast Cancer Cell Membranes.

    PubMed

    Sarkar, Mitul; Koland, John G

    2016-01-01

    The fluorescence recovery after photobleaching (FRAP) method is a straightforward means of assessing the diffusional mobility of membrane-associated proteins that is readily performed with current confocal microscopy instrumentation. We describe here the specific application of the FRAP method in characterizing the lateral diffusion of genetically encoded green fluorescence protein (GFP)-tagged plasma membrane receptor proteins. The method is exemplified in an examination of whether the previously observed segregation of the mammalian HER3 receptor protein in discrete plasma membrane microdomains results from its physical interaction with cellular entities that restrict its mobility. Our FRAP measurements of the diffusional mobility of GFP-tagged HER3 reporters expressed in MCF7 cultured breast cancer cells showed that despite the observed segregation of HER3 receptors within plasma membrane microdomains their diffusion on the macroscopic scale is not spatially restricted. Thus, in FRAP analyses of various HER3 reporters a near-complete recovery of fluorescence after photobleaching was observed, indicating that HER3 receptors are not immobilized by long-lived physical interactions with intracellular species. An examination of HER3 proteins with varying intracellular domain sequence truncations also indicated that a proposed formation of oligomeric HER3 networks, mediated by physical interactions involving specific HER3 intracellular domain sequences, either does not occur or does not significantly reduce HER3 mobility on the macroscopic scale.

  2. CDOM Sources and Photobleaching Control Quantum Yields for Oceanic DMS Photolysis.

    PubMed

    Galí, Martí; Kieber, David J; Romera-Castillo, Cristina; Kinsey, Joanna D; Devred, Emmanuel; Pérez, Gonzalo L; Westby, George R; Marrasé, Cèlia; Babin, Marcel; Levasseur, Maurice; Duarte, Carlos M; Agustí, Susana; Simó, Rafel

    2016-12-20

    Photolysis is a major removal pathway for the biogenic gas dimethylsulfide (DMS) in the surface ocean. Here we tested the hypothesis that apparent quantum yields (AQY) for DMS photolysis varied according to the quantity and quality of its photosensitizers, chiefly chromophoric dissolved organic matter (CDOM) and nitrate. AQY compiled from the literature and unpublished studies ranged across 3 orders of magnitude at the 330 nm reference wavelength. The smallest AQY(330) were observed in coastal waters receiving major riverine inputs of terrestrial CDOM (0.06-0.5 m 3 (mol quanta) -1 ). In open-ocean waters, AQY(330) generally ranged between 1 and 10 m 3 (mol quanta) -1 . The largest AQY(330), up to 34 m 3 (mol quanta) -1 ), were seen in the Southern Ocean potentially associated with upwelling. Despite the large AQY variability, daily photolysis rate constants at the sea surface spanned a smaller range (0.04-3.7 d -1 ), mainly because of the inverse relationship between CDOM absorption and AQY. Comparison of AQY(330) with CDOM spectral signatures suggests there is an interplay between CDOM origin (terrestrial versus marine) and photobleaching that controls variations in AQYs, with a secondary role for nitrate. Our results can be used for regional or large-scale assessment of DMS photolysis rates in future studies.

  3. Photobleaching Kinetics of Chromophoric Dissolved Organic Matter Derived from Mangrove Leaf Litter and Floating Sargassum Colonies

    EPA Science Inventory

    We examined the photoreactivity of chromophoric dissolved organic matter (CDOM) derived from Rhizophora mangle (red mangrove) leaf litter and floating Sargassum colonies as these marine plants can be important contributors to coastal and open ocean CDOM pools, respectively. Mangr...

  4. Spatio-seasonal variability of chromophoric dissolved organic matter absorption and responses to photobleaching in a large shallow temperate lake

    NASA Astrophysics Data System (ADS)

    Encina Aulló-Maestro, María; Hunter, Peter; Spyrakos, Evangelos; Mercatoris, Pierre; Kovács, Attila; Horváth, Hajnalka; Preston, Tom; Présing, Mátyás; Torres Palenzuela, Jesús; Tyler, Andrew

    2017-03-01

    The development and validation of remote-sensing-based approaches for the retrieval of chromophoric dissolved organic matter (CDOM) concentrations requires a comprehensive understanding of the sources and magnitude of variability in the optical properties of dissolved material within lakes. In this study, spatial and seasonal variability in concentration and composition of CDOM and the origin of its variation was studied in Lake Balaton (Hungary), a large temperate shallow lake in central Europe. In addition, we investigated the effect of photobleaching on the optical properties of CDOM through in-lake incubation experiments. There was marked variability throughout the year in CDOM absorption in Lake Balaton (aCDOM(440) = 0. 06-9.01 m-1). The highest values were consistently observed at the mouth of the main inflow (Zala River), which drains humic-rich material from the adjoining Kis-Balaton wetland, but CDOM absorption decreased rapidly towards the east where it was consistently lower and less variable than in the westernmost lake basins. The spectral slope parameter for the interval of 350-500 nm (SCDOM(350-500)) was more variable with increasing distance from the inflow (observed range 0.0161-0.0181 nm-1 for the mouth of the main inflow and 0.0158-0.0300 nm-1 for waters closer to the outflow). However, spatial variation in SCDOM was more constant exhibiting a negative correlation with aCDOM(440). Dissolved organic carbon (DOC) was strongly positively correlated with aCDOM(440) and followed a similar seasonal trend but it demonstrated more variability than either aCDOM or SCDOM with distance through the system. Photobleaching resulting from a 7-day exposure to natural solar UV radiation resulted in a marked decrease in allochthonous CDOM absorption (7.04 to 3.36 m-1, 42 % decrease). Photodegradation also resulted in an increase in the spectral slope coefficient of dissolved material.

  5. Single-molecule photobleaching reveals increased MET receptor dimerization upon ligand binding in intact cells

    PubMed Central

    2013-01-01

    Background The human receptor tyrosine kinase MET and its ligand hepatocyte growth factor/scatter factor are essential during embryonic development and play an important role during cancer metastasis and tissue regeneration. In addition, it was found that MET is also relevant for infectious diseases and is the target of different bacteria, amongst them Listeria monocytogenes that induces bacterial uptake through the surface protein internalin B. Binding of ligand to the MET receptor is proposed to lead to receptor dimerization. However, it is also discussed whether preformed MET dimers exist on the cell membrane. Results To address these issues we used single-molecule fluorescence microscopy techniques. Our photobleaching experiments show that MET exists in dimers on the membrane of cells in the absence of ligand and that the proportion of MET dimers increases significantly upon ligand binding. Conclusions Our results indicate that partially preformed MET dimers may play a role in ligand binding or MET signaling. The addition of the bacterial ligand internalin B leads to an increase of MET dimers which is in agreement with the model of ligand-induced dimerization of receptor tyrosine kinases. PMID:23731667

  6. A Light-Induced Reaction with Oxygen Leads to Chromophore Decomposition and Irreversible Photobleaching in GFP-Type Proteins.

    PubMed

    Grigorenko, Bella L; Nemukhin, Alexander V; Polyakov, Igor V; Khrenova, Maria G; Krylov, Anna I

    2015-04-30

    Photobleaching and photostability of proteins of the green fluorescent protein (GFP) family are crucially important for practical applications of these widely used biomarkers. On the basis of simulations, we propose a mechanism for irreversible bleaching in GFP-type proteins under intense light illumination. The key feature of the mechanism is a photoinduced reaction of the chromophore with molecular oxygen (O2) inside the protein barrel leading to the chromophore's decomposition. Using quantum mechanics/molecular mechanics (QM/MM) modeling we show that a model system comprising the protein-bound Chro(-) and O2 can be excited to an electronic state of the intermolecular charge-transfer (CT) character (Chro(•)···O2(-•)). Once in the CT state, the system undergoes a series of chemical reactions with low activation barriers resulting in the cleavage of the bridging bond between the phenolic and imidazolinone rings and disintegration of the chromophore.

  7. Improved murine glioma detection following modified diet and photobleaching of skin PpIX fluorescence

    NASA Astrophysics Data System (ADS)

    Gibbs, Summer L.; O'Hara, Julia A.; Hoopes, P. Jack; Pogue, Brian W.

    2007-02-01

    The Aminolevulinic Acid (ALA) - Protoporphyrin IX (PpIX) system is unique in the world of photosensitizers in that the prodrug ALA is enzymatically transformed via the tissue of interest into fluorescently detectable levels of PpIX. This system can be used to monitor cellular metabolism of tumor tissue for applications such as therapy monitoring. Detecting PpIX fluorescence noninvasively has proven difficult due to the high levels of PpIX produced in the skin compared to other tissue both with and without ALA administration. In the current study, methods to decrease skin PpIX autofluorescence and skin PpIX fluorescence following ALA administration have been examined. Use of a purified diet is found to decrease both skin PpIX autofluorescence and skin PpIX fluorescence following ALA administration, while addition of a broad spectrum antibiotic to the water shows little effect. Following ALA administration, improved brain tumor detection is seen when skin PpIX fluorescence is photobleached via blue light prior to transmission spectroscopic measurements of tumor bearing and control animals. Both of these methods to decrease skin PpIX autofluorescence and skin PpIX fluorescence following ALA administration are shown to have a large effect on the ability to detect tumor tissue PpIX fluorescence noninvasively in vivo.

  8. Instantaneous velocity measurement of AC electroosmotic flows by laser induced fluorescence photobleaching anemometer with high temporal resolution

    NASA Astrophysics Data System (ADS)

    Zhao, Wei; Yang, Fang; Qiao, Rui; Wang, Guiren; Rui Qiao Collaboration

    2015-11-01

    Understanding the instantaneous response of flows to applied AC electric fields may help understand some unsolved issues in induced-charge electrokinetics and enhance performance of microfluidic devices. Since currently available velocimeters have difficulty in measuring velocity fluctuations with frequency higher than 1 kHz, most experimental studies so far focus only on the average velocity measurement in AC electrokinetic flows. Here, we present measurements of AC electroosmotic flow (AC-EOF) response time in microchannels by a novel velocimeter with submicrometer spatial resolution and microsecond temporal resolution, i.e. laser-induced fluorescence photobleaching anemometer (LIFPA). Several parameters affecting the AC-EOF response time to the applied electric signal were investigated, i.e. channel length, transverse position and solution conductivity. The experimental results show that the EOF response time under a pulsed electric field decreases with the reduction of the microchannel length, distance between the detection position to the wall and the conductivity of the solution. This work could provide a new powerful tool to measure AC electrokinetics and enhance our understanding of AC electrokinetic flows.

  9. Analysis of caspase-3 in ASTC-a-1 cells treated with mitomycin C using acceptor photobleaching techniques

    NASA Astrophysics Data System (ADS)

    Wang, Huiying; Chen, Tongsheng; Sun, Lei

    2008-02-01

    Caspase-3 is a key activated death protease, which catalyzes the specific cleavage of many cellular proteins and induces DNA cleavage eventually. In this report, cells were treated with mitomycin C (MMC) at different concentration and its activity was detected by cell counting kit (CCK-8). Based on results of CCK-8, cells were treated with 10μg/mL MMC and Hoechst 33258 has been used to observe cell apoptosis. Fluorescence resonance energy transfer (FRET) and confocal microscopy have been used to the effect of MMC on the caspase3 activation in living cells. Human lung adenocarcinoma cells (ASTC-a-1) was transfected with plasmid SCAT3 (pSCAT3)/CKAR FRET receptor. Acceptor photobleaching techniques of FRET plasmid has been used to destruct fluorophore of cells stably expressing SCAT3 reporter on a fluorescence confocal microscope. The activity of caspase3 can be analyzed by FRET dynamics of SCAT3 in living cells. Our results show that MM C can induce ASTC-a-1 cell apoptosis through activation of caspase3.

  10. Microfiberoptic fluorescence photobleaching reveals size-dependent macromolecule diffusion in extracellular space deep in brain.

    PubMed

    Zador, Zsolt; Magzoub, Mazin; Jin, Songwan; Manley, Geoffrey T; Papadopoulos, Marios C; Verkman, A S

    2008-03-01

    Diffusion in brain extracellular space (ECS) is important for nonsynaptic intercellular communication, extracellular ionic buffering, and delivery of drugs and metabolites. We measured macromolecular diffusion in normally light-inaccessible regions of mouse brain by microfiberoptic epifluorescence photobleaching, in which a fiberoptic with a micron-size tip is introduced deep in brain tissue. In brain cortex, the diffusion of a noninteracting molecule [fluorescein isothiocyanate (FITC)-dextran, 70 kDa] was slowed 4.5 +/- 0.5-fold compared with its diffusion in water (D(o)/D), and was depth-independent down to 800 microm from the brain surface. Diffusion was significantly accelerated (D(o)/D of 2.9+/-0.3) in mice lacking the glial water channel aquaporin-4. FITC-dextran diffusion varied greatly in different regions of brain, with D(o)/D of 3.5 +/- 0.3 in hippocampus and 7.4 +/- 0.3 in thalamus. Remarkably, D(o)/D in deep brain was strongly dependent on solute size, whereas diffusion in cortex changed little with solute size. Mathematical modeling of ECS diffusion required nonuniform ECS dimensions in deep brain, which we call "heterometricity," to account for the size-dependent diffusion. Our results provide the first data on molecular diffusion in ECS deep in brain in vivo and demonstrate previously unrecognized hindrance and heterometricity for diffusion of large macromolecules in deep brain.

  11. Analysis of the diffusion of Ras2 in Saccharomyces cerevisiae using fluorescence recovery after photobleaching

    NASA Astrophysics Data System (ADS)

    Vinnakota, Kalyan C.; Mitchell, David A.; Deschenes, Robert J.; Wakatsuki, Tetsuro; Beard, Daniel A.

    2010-06-01

    Binding, lateral diffusion and exchange are fundamental dynamic processes involved in protein association with cellular membranes. In this study, we developed numerical simulations of lateral diffusion and exchange of fluorophores in membranes with arbitrary bleach geometry and exchange of the membrane-localized fluorophore with the cytosol during fluorescence recovery after photobleaching (FRAP) experiments. The model simulations were used to design FRAP experiments with varying bleach region sizes on plasma membrane-localized wild-type GFP-Ras2 with a dual lipid anchor and mutant GFP-Ras2C318S with a single lipid anchor in live yeast cells to investigate diffusional mobility and the presence of any exchange processes operating in the time scale of our experiments. Model parameters estimated using data from FRAP experiments with a 1 µm × 1 µm bleach region-of-interest (ROI) and a 0.5 µm × 0.5 µm bleach ROI showed that GFP-Ras2, single or dual lipid modified, diffuses as single species with no evidence of exchange with a cytoplasmic pool. This is the first report of Ras2 mobility in the yeast plasma membrane. The methods developed in this study are generally applicable for studying diffusion and exchange of membrane-associated fluorophores using FRAP on commercial confocal laser scanning microscopes.

  12. Variations in gap junctional intercellular communication and connexin expression in fibroblasts derived from keloid and hypertrophic scars.

    PubMed

    Lu, Feng; Gao, JianHua; Ogawa, Rei; Hyakusoku, Hiko

    2007-03-01

    Expression of connexins and other constituent proteins of gap junctions along with gap junctional intercellular communication are involved in cellular development and differentiation processes. In addition, an increasing number of hereditary skin disorders appear to be linked to connexins. Therefore, in this report, the authors studied in vitro gap junctional intercellular communication function and connexin expression in fibroblasts derived from keloid and hypertrophic scar patients. Fibroblasts harvested from each of six keloid and hypertrophic scar patients were used for this study. Gap junctional intercellular communication function was investigated using the gap fluorescence recovery after photobleaching method, and expression of connexin proteins was studied using quantitative confocal microscopic analyses. Compared with normal skin, a decreased level of gap junctional intercellular communication was seen in fibroblasts derived from hypertrophic scar tissue, whereas an extremely low gap junctional intercellular communication level was detected in fibroblasts derived from keloid tissue. We also detected little connexin 43 (Cx43) protein localized in fibroblasts derived from keloids. Moreover, Cx43 protein levels were much lower in fibroblasts derived from hypertrophic scars than in those derived from normal skin. The authors' data suggest that the loss of gap junctional intercellular communication and connexin expression may affect intercellular recognition and thus break the proliferation and apoptosis balance in fibroblasts derived from keloid and hypertrophic scar tissue.

  13. Subsurface femtosecond tissue alteration: selectively photobleaching macular degeneration pigments in near retinal contact.

    PubMed

    Manevitch, Zakhariya; Lewis, Aaron; Levy, Carol; Zeira, Evelyne; Banin, Eyal; Manevitch, Alexandra; Khatchatouriants, Artium; Pe'er, Jacob; Galun, Eithan; Hemo, Itzhak

    2012-06-14

    This paper uses advances in the ultrafast manipulation of light to address a general need in medicine for a clinical approach that can provide a solution to a variety of disorders requiring subsurface tissue manipulation with ultralow collateral damage. Examples are age-related macular degeneration (AMD), fungal infections, tumors surrounded by overlying tissue, cataracts, etc. Although lasers have revolutionized the use of light in clinical settings, most lasers employed in medicine cannot address such problems of depth-selective tissue manipulation. This arises from the fact that they are mostly based on one photon based laser tissue interactions that provide a cone of excitation where the energy density is sufficiently high to excite heat or fluorescence in the entire cone. Thus, it is difficult to excite a specific depth of a tissue without affecting the overlying surface. However, the advent of femtosecond (fs) lasers has caused a revolution in multiphoton microscopy (Zipfel et al. Nat. Biotechnol. 2003, 21, 1369-1377; Denk et al. Science 1990, 248, 73-76) and fabrication (Kawata et al. Nature 2001, 412, 697-698). With such lasers, the photon energy density is only high enough for multiphoton processes in the focal volume, and this opens a new direction to address subsurface tissue manipulation. Here we show in an AMD animal model, Ccr2 KO knockout mutant mice, noninvasive, selective fs two-photon photobleaching of pigments associated with AMD that accumulate under and in ultraclose proximity to the overlying retina. Pathological evidence is presented that indicates the lack of collateral damage to the overlying retina or other surrounding tissue.

  14. Monochrome Multiplexing in Polymerase Chain Reaction by Photobleaching of Fluorogenic Hydrolysis Probes.

    PubMed

    Schuler, Friedrich; Trotter, Martin; Zengerle, Roland; von Stetten, Felix

    2016-03-01

    Multiplexing in polymerase chain reaction (PCR) is a technique widely used to save cost and sample material and to increase sensitivity compared to distributing a sample to several singleplex reactions. One of the most common methods to detect the different amplification products is the use of fluorogenic probes that emit at different wavelengths (colors). To reduce the number of detection channels, several methods for monochrome multiplexing have been suggested. However, they pose restrictions to the amplifiable target length, the sequence, or the melting temperature. To circumvent these limitations, we suggest a novel approach that uses different fluorophores with the same emission maximum. Discrimination is achieved by their different fluorescence stability during photobleaching. Atto488 (emitting at the same wavelength as 6-carboxyfluorescein, FAM) and Atto467N (emitting at the same wavelength as cyanine 5, Cy5) were found to bleach significantly less than FAM and Cy5; i.e., the final fluorescence of Atto dyes was more than tripled compared to FAM and Cy5. We successfully applied this method by performing a 4-plex PCR targeting antibiotic resistance genes in S. aureus using only 2 color channels. Confidence of discrimination between the targets was >99.9% at high copy initial copy numbers of 100 000 copies. Cases where both targets were present could be discriminated with equal confidence for Cy5 channel and reduced levels of confidence (>68%) for FAM channel. Moreover, a 2-plex digital PCR reaction in 1 color channel was shown. In the future, the degree of multiplexing may be increased by adding fluorogenic probe pairs with other emission wavelengths. The method may also be applied to other probe and assay formats, such as Förster resonance energy transfer (FRET) probes and immunoassays.

  15. Reducing the photo-bleaching effect of a new europium complex embedded in styrene butadiene copolymer

    NASA Astrophysics Data System (ADS)

    Jiménez, G. Lesly; Reyes-Rodríguez, J. L.; Padilla, Isela; Alarcón-Flores, G.; Falcony, C.

    2018-02-01

    A highly luminescent europium complex obtained with two different ligands, succinimide (SI) and 2-thenoyltrifluoroacetone (TTA) , was synthetized with different TTA concentrations. The photoluminescence (PL) emission from these materials corresponds to the characteristic inter-electronic energy level transitions of the Eu3+ ions. However, the excitation spectrum is strongly dependent on the presence of TTA, having an optimum response when 0.75 mmol of this compound is added to the EuL3(H2O)3 complex. The quantum yield obtained by these powders were around 72 % ± 1.7 % indicating an optimum sensitization of these complex. The EuL3 TTA complex with the best PL properties was embedded in a styrene butadiene copolymer (SBC) film, produced by the drop casting method, obtaining similar PL behavior at different concentrations, the highest intensity was observed at 1.2% (w/v) of EuL3 TTA complex and the quantum yield of these composite films was 60.5 % ± 2 % . These films were exposed to continuous UV irradiation and after 141 h no photo-bleaching effect was observed in contrast with the EuL3 TTA complex that exhibited a noticeable photoluminescence intensity degradation at much shorter exposure times. Both the Eu-complexes and the composite films were characterized by FT-IR, XRD, SEM and fluorescence spectroscopy.

  16. Quantification of Lacunar-Canalicular Interstitial Fluid Flow Through Computational Modeling of Fluorescence Recovery After Photobleaching.

    PubMed

    Kwon, Ronald Y; Frangos, John A

    2010-09-01

    Skeletal adaptation to mechanical loading has been widely hypothesized to involve the stimulation of osteocytes by interstitial fluid flow (IFF). However, direct investigation of this hypothesis has been difficult due in large part to the inability to directly measure IFF velocities within the lacunar-canalicular system. Measurements of fluorescence recovery after photobleaching (FRAP) within individual lacunae could be used to quantify lacunar-canalicular IFF when combined with mathematical modeling. In this study, we used a computational transport model to characterize the relationship between flow frequency (0.5-10 Hz), peak flow velocity (0-300 μm/s), tracer diffusion coefficient (100-300 μm(2)/s), and transport enhancement (i.e., (k/k(0)) - 1, where k and k(0) are the transport rates in the presence/absence of flow) during lacunar FRAP investigations. We show that this relationship is well described by a simple power law with frequency-dependent coefficients, and is relatively insensitive to variations in lacunar geometry. Using this power law relationship, we estimated peak IFF velocities in hindlimb mice subjected to intramedullary pressurization using values of k and k(0) previously obtained from ex vivo lacunar FRAP investigations. Together, our findings suggest that skeletal adaptation in hindlimb suspended mice subjected to dynamic intramedullary pressure occurred in the presence of IFF at levels associated with physiological loading.

  17. Quantification of Lacunar–Canalicular Interstitial Fluid Flow Through Computational Modeling of Fluorescence Recovery After Photobleaching

    PubMed Central

    Kwon, Ronald Y.; Frangos, John A.

    2010-01-01

    Skeletal adaptation to mechanical loading has been widely hypothesized to involve the stimulation of osteocytes by interstitial fluid flow (IFF). However, direct investigation of this hypothesis has been difficult due in large part to the inability to directly measure IFF velocities within the lacunar–canalicular system. Measurements of fluorescence recovery after photobleaching (FRAP) within individual lacunae could be used to quantify lacunar–canalicular IFF when combined with mathematical modeling. In this study, we used a computational transport model to characterize the relationship between flow frequency (0.5–10 Hz), peak flow velocity (0–300 μm/s), tracer diffusion coefficient (100–300 μm2/s), and transport enhancement (i.e., (k/k0) − 1, where k and k0 are the transport rates in the presence/absence of flow) during lacunar FRAP investigations. We show that this relationship is well described by a simple power law with frequency-dependent coefficients, and is relatively insensitive to variations in lacunar geometry. Using this power law relationship, we estimated peak IFF velocities in hindlimb mice subjected to intramedullary pressurization using values of k and k0 previously obtained from ex vivo lacunar FRAP investigations. Together, our findings suggest that skeletal adaptation in hindlimb suspended mice subjected to dynamic intramedullary pressure occurred in the presence of IFF at levels associated with physiological loading. PMID:21076644

  18. An improved strip FRAP method for estimating diffusion coefficients: correcting for the degree of photobleaching.

    PubMed

    Yang, J; Köhler, K; Davis, D M; Burroughs, N J

    2010-06-01

    Fluorescence recovery after photobleaching is a widely established method for the estimation of diffusion coefficients, strip bleaching with an associated recovery curve analysis being one of the simplest techniques. However, its implementation requires near 100% bleaching in the region of interest with negligible fluorescence loss outside, both constraints being hard to achieve concomitantly for fast diffusing molecules. We demonstrate that when these requirements are not met there is an error in the estimation of the diffusion coefficient D, either an under- or overestimation depending on which assumption is violated the most. We propose a simple modification to the recovery curve analysis incorporating the concept of the relative bleached mass m giving a revised recovery time parametrization tau=m(2)w(2)/4piD for a strip of width w. This modified model removes the requirement of 100% bleaching in the region of interest and allows for limited diffusion of the fluorophore during bleaching. We validate our method by estimating the (volume) diffusion coefficient of FITC-labelled IgG in 60% glycerol solution, D= 4.09 +/- 0.21 microm(2) s(-1), and the (surface) diffusion coefficient of a green-fluorescent protein-tagged class I MHC protein expressed at the surface of a human B cell line, D= 0.32 +/- 0.03 microm(2) s(-1) for a population of cells.

  19. Photophysics of Thermally-Assisted Photobleaching in "Giant" Quantum Dots Revealed in Single Nanocrystals.

    PubMed

    Orfield, Noah J; Majumder, Somak; McBride, James R; Yik-Ching Koh, Faith; Singh, Ajay; Bouquin, Sarah J; Casson, Joanna L; Johnson, Alex D; Sun, Liuyang; Li, Xiaoqin; Shih, Chih-Kang; Rosenthal, Sandra J; Hollingsworth, Jennifer A; Htoon, Han

    2018-05-07

    Quantum dots (QDs) are steadily being implemented as down-conversion phosphors in market-ready display products to enhance color rendering, brightness, and energy efficiency. However, for adequate longevity, QDs must be encased in a protective barrier that separates them from ambient oxygen and humidity, and device architectures are designed to avoid significant heating of the QDs as well as direct contact between the QDs and the excitation source. In order to increase the utility of QDs in display technologies and to extend their usefulness to more demanding applications as, for example, alternative phosphors for solid-state lighting (SSL), QDs must retain their photoluminescence emission properties over extended periods of time under conditions of high temperature and high light flux. Doing so would simplify the fabrication costs for QD display technologies and enable QDs to be used as down-conversion materials in light-emitting diodes for SSL, where direct-on-chip configurations expose the emitters to temperatures approaching 100 °C and to photon fluxes from 0.1 W/mm 2 to potentially 10 W/mm 2 . Here, we investigate the photobleaching processes of single QDs exposed to controlled temperature and photon flux. In particular, we investigate two types of room-temperature-stable core/thick-shell QDs, known as "giant" QDs for which shell growth is conducted using either a standard layer-by-layer technique or by a continuous injection method. We determine the mechanistic pathways responsible for thermally-assisted photodegradation, distinguishing effects of hot-carrier trapping and QD charging. The findings presented here will assist in the further development of advanced QD heterostructures for maximum device lifetime stability.

  20. Mobility of adsorbed Cry1Aa insecticidal toxin from Bacillus thuringiensis (Bt) on montmorillonite measured by fluorescence recovery after photobleaching (FRAP)

    NASA Astrophysics Data System (ADS)

    Helassa, Nordine; Daudin, Gabrielle; Noinville, Sylvie; Janot, Jean-Marc; Déjardin, Philippe; Staunton, Siobhán; Quiquampoix, Hervé

    2010-06-01

    The insecticidal toxins produced by genetically modified Bt crops are introduced into soil through root exudates and tissue decomposition and adsorb readily on soil components, especially on clays. This immobilisation and the consequent concentration of the toxins in "hot spots" could increase the exposure of soil organisms. Whereas the effects on non-target organisms are well documented, few studies consider the migration of the toxin in soil. In this study, the residual mobility of Bt Cry1Aa insecticidal toxin adsorbed on montmorillonite was assessed using fluorescence recovery after photobleaching (FRAP). This technique, which is usually used to study dynamics of cytoplasmic and membrane molecules in live cells, was applied for the first time to a protein adsorbed on a finely divided swelling clay mineral, montmorillonite. No mobility of adsorbed toxin was observed at any pH and at different degrees of surface saturation.

  1. Actin dynamics at the living cell submembrane imaged by total internal reflection fluorescence photobleaching.

    PubMed Central

    Sund, S E; Axelrod, D

    2000-01-01

    Although reversible chemistry is crucial to dynamical processes in living cells, relatively little is known about relevant chemical kinetic rates in vivo. Total internal reflection/fluorescence recovery after photobleaching (TIR/FRAP), an established technique previously demonstrated to measure reversible biomolecular kinetic rates at surfaces in vitro, is extended here to measure reversible biomolecular kinetic rates of actin at the cytofacial (subplasma membrane) surface of living cells. For the first time, spatial imaging (with a charge-coupled device camera) is used in conjunction with TIR/FRAP. TIR/FRAP imaging produces both spatial maps of kinetic parameters (off-rates and mobile fractions) and estimates of kinetic correlation distances, cell-wide kinetic gradients, and dependences of kinetic parameters on initial fluorescence intensity. For microinjected rhodamine actin in living cultured smooth muscle (BC3H1) cells, the unbinding rate at or near the cytofacial surface of the plasma membrane (averaged over the entire cell) is measured at 0.032 +/- 0.007 s(-1). The corresponding rate for actin marked by microinjected rhodamine phalloidin is very similar, 0.033 +/- 0.013 s(-1), suggesting that TIR/FRAP is reporting the dynamics of entire filaments or protofilaments. For submembrane fluorescence-marked actin, the intensity, off-rate, and mobile fraction show a positive correlation over a characteristic distance of 1-3 microm and a negative correlation over larger distances greater than approximately 7-14 microm. Furthermore, the kinetic parameters display a statistically significant cell-wide gradient, with the cell having a "fast" and "slow" end with respect to actin kinetics. PMID:10969025

  2. Anomalous diffusion and dynamics of fluorescence recovery after photobleaching in the random-comb model

    NASA Astrophysics Data System (ADS)

    Yuste, S. B.; Abad, E.; Baumgaertner, A.

    2016-07-01

    We address the problem of diffusion on a comb whose teeth display varying lengths. Specifically, the length ℓ of each tooth is drawn from a probability distribution displaying power law behavior at large ℓ ,P (ℓ ) ˜ℓ-(1 +α ) (α >0 ). To start with, we focus on the computation of the anomalous diffusion coefficient for the subdiffusive motion along the backbone. This quantity is subsequently used as an input to compute concentration recovery curves mimicking fluorescence recovery after photobleaching experiments in comblike geometries such as spiny dendrites. Our method is based on the mean-field description provided by the well-tested continuous time random-walk approach for the random-comb model, and the obtained analytical result for the diffusion coefficient is confirmed by numerical simulations of a random walk with finite steps in time and space along the backbone and the teeth. We subsequently incorporate retardation effects arising from binding-unbinding kinetics into our model and obtain a scaling law characterizing the corresponding change in the diffusion coefficient. Finally, we show that recovery curves obtained with the help of the analytical expression for the anomalous diffusion coefficient cannot be fitted perfectly by a model based on scaled Brownian motion, i.e., a standard diffusion equation with a time-dependent diffusion coefficient. However, differences between the exact curves and such fits are small, thereby providing justification for the practical use of models relying on scaled Brownian motion as a fitting procedure for recovery curves arising from particle diffusion in comblike systems.

  3. Minimizing photodecomposition of flavin adenine dinucleotide fluorescence by the use of pulsed LEDs.

    PubMed

    Rösner, J; Liotta, A; Angamo, E A; Spies, C; Heinemann, U; Kovács, R

    2016-11-01

    Dynamic alterations in flavin adenine dinucleotide (FAD) fluorescence permit insight into energy metabolism-dependent changes of intramitochondrial redox potential. Monitoring FAD fluorescence in living tissue is impeded by photobleaching, restricting the length of microfluorimetric recordings. In addition, photodecomposition of these essential electron carriers negatively interferes with energy metabolism and viability of the biological specimen. Taking advantage of pulsed LED illumination, here we determined the optimal excitation settings giving the largest fluorescence yield with the lowest photobleaching and interference with metabolism in hippocampal brain slices. The effects of FAD bleaching on energy metabolism and viability were studied by monitoring tissue pO 2 , field potentials and changes in extracellular potassium concentration ([K + ] o ). Photobleaching with continuous illumination consisted of an initial exponential decrease followed by a nearly linear decay. The exponential decay was significantly decelerated with pulsed illumination. Pulse length of 5 ms was sufficient to reach a fluorescence output comparable to continuous illumination, whereas further increasing duration increased photobleaching. Similarly, photobleaching increased with shortening of the interpulse interval. Photobleaching was partially reversible indicating the existence of a transient nonfluorescent flavin derivative. Pulsed illumination decreased FAD photodecomposition, improved slice viability and reproducibility of stimulus-induced FAD, field potential, [K + ] o and pO 2 changes as compared to continuous illumination. © 2016 The Authors Journal of Microscopy © 2016 Royal Microscopical Society.

  4. The molecular basis of the solution properties of hyaluronan investigated by confocal fluorescence recovery after photobleaching.

    PubMed Central

    Gribbon, P; Heng, B C; Hardingham, T E

    1999-01-01

    Hyaluronan (HA) is a highly hydrated polyanion, which is a network-forming and space-filling component in the extracellular matrix of animal tissues. Confocal fluorescence recovery after photobleaching (confocal-FRAP) was used to investigate intramolecular hydrogen bonding and electrostatic interactions in hyaluronan solutions. Self and tracer lateral diffusion coefficients within hyaluronan solutions were measured over a wide range of concentrations (c), with varying electrolyte and at neutral and alkaline pH. The free diffusion coefficient of fluoresceinamine-labeled HA of 500 kDa in PBS was 7.9 x 10(-8) cm(2) s(-1) and of 830 kDa HA was 5.6 x 10(-8) cm(2) s(-1). Reductions in self- and tracer-diffusion with c followed a stretched exponential model. Electrolyte-induced polyanion coil contraction and destiffening resulted in a 2.8-fold increase in self-diffusion between 0 and 100 mM NaCl. Disruption of hydrogen bonds by strong alkali (0.5 M NaOH) resulted in further larger increases in self- and tracer-diffusion coefficients, consistent with a more dynamic and permeable network. Concentrated hyaluronan solution properties were attributed to hydrodynamic and entanglement interactions between domains. There was no evidence of chain-chain associations. At physiological electrolyte concentration and pH, the greatest contribution to the intrinsic stiffness of hyaluronan appeared to be due to hydrogen bonds between adjacent saccharides. PMID:10512840

  5. Photobleaching effect in azo-dye containing epoxy resin films: the potentiality of carbon nanotubes as azo-dye dispensers

    NASA Astrophysics Data System (ADS)

    Díaz Costanzo, Guadalupe; Goyanes, Silvia; Ledesma, Silvia

    2015-04-01

    Azo-dye molecules may suffer from bleaching under certain illumination conditions. When this photoinduced process occurs, it generates an irreversible effect that is characterized by the loss of absorption of the dye molecule. Moreover, the well-known isomerization of azodye molecules does not occur anymore. In this work it is shown how the addition of a small amount of multi-walled carbon nanotubes (MWCNTs) helps to decrease the bleaching effect in a photosensitive guest-host azo-polymer film. Two different systems were fabricated using an epoxy resin as polymer matrix. An azo-dye, Disperse Orange 3, was used as photosensitive material in both systems and MWCNTs were added into one of them. The optical response of the polymeric systems was studied considering the degree of photoinduced birefringence. Photobleaching of the azo-dye was observed in all cases however, the effect is lower for the composite material containing 0.2 wt % MWCNTs. The weak interaction between MWCNTs and dye molecules is less favorable when the material is heated. The optical behavior of the heated composite material suggests that carbon nanotubes can be potentially used as azo dye dispensers. The results are interpreted in terms of the non-covalent interaction between azo-dye molecules and MWCNTs.

  6. Assessing cellular efficacy of bromodomain inhibitors using fluorescence recovery after photobleaching

    PubMed Central

    2014-01-01

    Background Acetylation of lysine residues in histone tails plays an important role in the regulation of gene transcription. Bromdomains are the readers of acetylated histone marks, and, consequently, bromodomain-containing proteins have a variety of chromatin-related functions. Moreover, they are increasingly being recognised as important mediators of a wide range of diseases. The first potent and selective bromodomain inhibitors are beginning to be described, but the diverse or unknown functions of bromodomain-containing proteins present challenges to systematically demonstrating cellular efficacy and selectivity for these inhibitors. Here we assess the viability of fluorescence recovery after photobleaching (FRAP) assays as a target agnostic method for the direct visualisation of an on-target effect of bromodomain inhibitors in living cells. Results Mutation of a conserved asparagine crucial for binding to acetylated lysines in the bromodomains of BRD3, BRD4 and TRIM24 all resulted in reduction of FRAP recovery times, indicating loss of or significantly reduced binding to acetylated chromatin, as did the addition of known inhibitors. Significant differences between wild type and bromodomain mutants for ATAD2, BAZ2A, BRD1, BRD7, GCN5L2, SMARCA2 and ZMYND11 required the addition of the histone deacetylase inhibitor suberoylanilide hydroxamic acid (SAHA) to amplify the binding contribution of the bromodomain. Under these conditions, known inhibitors decreased FRAP recovery times back to mutant control levels. Mutation of the bromodomain did not alter FRAP recovery times for full-length CREBBP, even in the presence of SAHA, indicating that other domains are primarily responsible for anchoring CREBBP to chromatin. However, FRAP assays with multimerised CREBBP bromodomains resulted in a good assay to assess the efficacy of bromodomain inhibitors to this target. The bromodomain and extraterminal protein inhibitor PFI-1 was inactive against other bromodomain targets

  7. Quantitative fluorescence loss in photobleaching for analysis of protein transport and aggregation

    PubMed Central

    2012-01-01

    Background Fluorescence loss in photobleaching (FLIP) is a widely used imaging technique, which provides information about protein dynamics in various cellular regions. In FLIP, a small cellular region is repeatedly illuminated by an intense laser pulse, while images are taken with reduced laser power with a time lag between the bleaches. Despite its popularity, tools are lacking for quantitative analysis of FLIP experiments. Typically, the user defines regions of interest (ROIs) for further analysis which is subjective and does not allow for comparing different cells and experimental settings. Results We present two complementary methods to detect and quantify protein transport and aggregation in living cells from FLIP image series. In the first approach, a stretched exponential (StrExp) function is fitted to fluorescence loss (FL) inside and outside the bleached region. We show by reaction–diffusion simulations, that the StrExp function can describe both, binding/barrier–limited and diffusion-limited FL kinetics. By pixel-wise regression of that function to FL kinetics of enhanced green fluorescent protein (eGFP), we determined in a user-unbiased manner from which cellular regions eGFP can be replenished in the bleached area. Spatial variation in the parameters calculated from the StrExp function allow for detecting diffusion barriers for eGFP in the nucleus and cytoplasm of living cells. Polyglutamine (polyQ) disease proteins like mutant huntingtin (mtHtt) can form large aggregates called inclusion bodies (IB’s). The second method combines single particle tracking with multi-compartment modelling of FL kinetics in moving IB’s to determine exchange rates of eGFP-tagged mtHtt protein (eGFP-mtHtt) between aggregates and the cytoplasm. This method is self-calibrating since it relates the FL inside and outside the bleached regions. It makes it therefore possible to compare release kinetics of eGFP-mtHtt between different cells and experiments. Conclusions We

  8. Photophysical investigation of cyano-substituted terrylenediimide derivatives.

    PubMed

    Kennes, Koen; Baeten, Yannick; Vosch, Tom; Sempels, Wouter; Yordanov, Stoyan; Stappert, Sebastian; Chen, Long; Müllen, Klaus; Hofkens, Johan; Van der Auweraer, Mark; Fron, Eduard

    2014-12-18

    Two new terrylenediimide (TDI) chromophores with cyano substituents in the bay and core area (BCN-TDI and OCN-TDI, respectively) have been characterized by a wide range of techniques, and their applicability for stimulated emission depletion (STED) microscopy has been tested. By cyano substitution an increase of the fluorescence quantum yield and a decrease of the nonradiative rate constant is achieved and attributed to a reduced charge-transfer character of the excited state due to a lower electron density of the TDI core. For BCN-TDI, the substitution in the bay area induces a strong torsional twist in the molecule which, similar to phenoxy bay-perylenediimide (PDI), has a strong effect on the fluorescence lifetime but appears to prevent the aggregation that is observed for OCN-TDI. The single-molecule photobleaching stability of BCN- and OCN-TDI is lower than that of a reference TDI without cyano substitution (C7-TDI), although less so for OCN-TDI. The photophysical properties of the excited singlet state are only slightly influenced by the cyano groups. The observed intense stimulated emission, the pump-dump-probe experiments, and STED single-molecule imaging indicate that STED experiments with the cyano-substituted TDIs are possible. However, because of aggregation and more efficient photobleaching, the performance of BCN- and OCN-TDI is worse than that of the reference compound without cyano groups (C7-TDI). Bay-substituted TDIs are less suitable for STED microscopy.

  9. Improving the photostability of bright monomeric orange and red fluorescent proteins.

    PubMed

    Shaner, Nathan C; Lin, Michael Z; McKeown, Michael R; Steinbach, Paul A; Hazelwood, Kristin L; Davidson, Michael W; Tsien, Roger Y

    2008-06-01

    All organic fluorophores undergo irreversible photobleaching during prolonged illumination. Although fluorescent proteins typically bleach at a substantially slower rate than many small-molecule dyes, in many cases the lack of sufficient photostability remains an important limiting factor for experiments requiring large numbers of images of single cells. Screening methods focusing solely on brightness or wavelength are highly effective in optimizing both properties, but the absence of selective pressure for photostability in such screens leads to unpredictable photobleaching behavior in the resulting fluorescent proteins. Here we describe an assay for screening libraries of fluorescent proteins for enhanced photostability. With this assay, we developed highly photostable variants of mOrange (a wavelength-shifted monomeric derivative of DsRed from Discosoma sp.) and TagRFP (a monomeric derivative of eqFP578 from Entacmaea quadricolor) that maintain most of the beneficial qualities of the original proteins and perform as reliably as Aequorea victoria GFP derivatives in fusion constructs.

  10. Self-Assembly of Electron Donor-Acceptor-Based Carbazole Derivatives: Novel Fluorescent Organic Nanoprobes for Both One- and Two-Photon Cellular Imaging.

    PubMed

    Zhang, Jinfeng; Chen, Wencheng; Kalytchuk, Sergii; Li, King Fai; Chen, Rui; Adachi, Chihaya; Chen, Zhan; Rogach, Andrey L; Zhu, Guangyu; Yu, Peter K N; Zhang, Wenjun; Cheah, Kok Wai; Zhang, Xiaohong; Lee, Chun-Sing

    2016-05-11

    In this study, we report fluorescent organic nanoprobes with intense blue, green, and orange-red emissions prepared by self-assembling three carbazole derivatives into nanorods/nanoparticles. The three compounds consist of two or four electron-donating carbazole groups linked to a central dicyanobenzene electron acceptor. Steric hindrance from the carbazole groups leads to noncoplanar 3D molecular structures favorable to fluorescence in the solid state, while the donor-acceptor structures endow the molecules with good two-photon excited emission properties. The fluorescent organic nanoprobes exhibit good water dispersibility, low cytotoxicity, superior resistance against photodegradation and photobleaching. Both one- and two-photon fluorescent imaging were shown in the A549 cell line. Two-photon fluorescence imaging with the fluorescent probes was demonstrated to be more effective in visualizing and distinguishing cellular details compared to conventional one-photon fluorescence imaging.

  11. Quantifying transient binding of ISWI chromatin remodelers in living cells by pixel-wise photobleaching profile evolution analysis.

    PubMed

    Erdel, Fabian; Rippe, Karsten

    2012-11-20

    Interactions between nuclear proteins and chromatin frequently occur on the time scale of seconds and below. These transient binding events are important for the fast identification of target sites as concluded from our previous analysis of the human chromatin remodelers Snf2H and Snf2L from the imitation switch (ISWI) family. Both ATP-driven molecular motor proteins are able to translocate nucleosomes along the DNA and appear to exert this activity only on a small number of nucleosomes to which they bind more tightly. For mechanistic studies, one needs to distinguish such translocation reactions or other long-lived interactions associated with conformational changes and/or ATP hydrolysis from nonproductive chromatin sampling during target search. These processes can be separated by measuring the duration of nucleosome binding with subsecond time resolution. To reach this goal, we have developed a fluorescence bleaching technique termed pixel-wise photobleaching profile evolution analysis (3PEA). It exploits the inherent time structure of confocal microscopy images and yields millisecond resolution. 3PEA represents a generally applicable approach to quantitate transient chromatin interactions in the 2- to 500-ms time regime within only ∼1 s needed for a measurement. The green autofluorescent protein (GFP)-tagged Snf2H and Snf2L and the inactive Snf2L+13 splice variant were studied by 3PEA in comparison to the isolated GFP or red autofluorescent protein and a GFP pentamer. Our results reveal that the residence time for transient chromatin binding of Snf2H and Snf2L is <2 ms, and strongly support the view that ISWI-type remodelers are only rarely active in unperturbed cells during G1 phase.

  12. Random-Walk Model of Diffusion in Three Dimensions in Brain Extracellular Space: Comparison with Microfiberoptic Photobleaching Measurements

    PubMed Central

    Jin, Songwan; Zador, Zsolt; Verkman, A. S.

    2008-01-01

    Diffusion through the extracellular space (ECS) in brain is important in drug delivery, intercellular communication, and extracellular ionic buffering. The ECS comprises ∼20% of brain parenchymal volume and contains cell-cell gaps ∼50 nm. We developed a random-walk model to simulate macromolecule diffusion in brain ECS in three dimensions using realistic ECS dimensions. Model inputs included ECS volume fraction (α), cell size, cell-cell gap geometry, intercellular lake (expanded regions of brain ECS) dimensions, and molecular size of the diffusing solute. Model output was relative solute diffusion in water versus brain ECS (Do/D). Experimental Do/D for comparison with model predictions was measured using a microfiberoptic fluorescence photobleaching method involving stereotaxic insertion of a micron-size optical fiber into mouse brain. Do/D for the small solute calcein in different regions of brain was in the range 3.0–4.1, and increased with brain cell swelling after water intoxication. Do/D also increased with increasing size of the diffusing solute, particularly in deep brain nuclei. Simulations of measured Do/D using realistic α, cell size and cell-cell gap required the presence of intercellular lakes at multicell contact points, and the contact length of cell-cell gaps to be least 50-fold smaller than cell size. The model accurately predicted Do/D for different solute sizes. Also, the modeling showed unanticipated effects on Do/D of changing ECS and cell dimensions that implicated solute trapping by lakes. Our model establishes the geometric constraints to account quantitatively for the relatively modest slowing of solute and macromolecule diffusion in brain ECS. PMID:18469079

  13. Random-walk model of diffusion in three dimensions in brain extracellular space: comparison with microfiberoptic photobleaching measurements.

    PubMed

    Jin, Songwan; Zador, Zsolt; Verkman, A S

    2008-08-01

    Diffusion through the extracellular space (ECS) in brain is important in drug delivery, intercellular communication, and extracellular ionic buffering. The ECS comprises approximately 20% of brain parenchymal volume and contains cell-cell gaps approximately 50 nm. We developed a random-walk model to simulate macromolecule diffusion in brain ECS in three dimensions using realistic ECS dimensions. Model inputs included ECS volume fraction (alpha), cell size, cell-cell gap geometry, intercellular lake (expanded regions of brain ECS) dimensions, and molecular size of the diffusing solute. Model output was relative solute diffusion in water versus brain ECS (D(o)/D). Experimental D(o)/D for comparison with model predictions was measured using a microfiberoptic fluorescence photobleaching method involving stereotaxic insertion of a micron-size optical fiber into mouse brain. D(o)/D for the small solute calcein in different regions of brain was in the range 3.0-4.1, and increased with brain cell swelling after water intoxication. D(o)/D also increased with increasing size of the diffusing solute, particularly in deep brain nuclei. Simulations of measured D(o)/D using realistic alpha, cell size and cell-cell gap required the presence of intercellular lakes at multicell contact points, and the contact length of cell-cell gaps to be least 50-fold smaller than cell size. The model accurately predicted D(o)/D for different solute sizes. Also, the modeling showed unanticipated effects on D(o)/D of changing ECS and cell dimensions that implicated solute trapping by lakes. Our model establishes the geometric constraints to account quantitatively for the relatively modest slowing of solute and macromolecule diffusion in brain ECS.

  14. Time-resolved fluorescence microscopy to study biologically related applications using sol-gel derived and cellular media

    NASA Astrophysics Data System (ADS)

    Toury, Marion; Chandler, Lin; Allison, Archie; Campbell, David; McLoskey, David; Holmes-Smith, A. Sheila; Hungerford, Graham

    2011-03-01

    Fluorescence microscopy provides a non-invasive means for visualising dynamic protein interactions. As well as allowing the calculation of kinetic processes via the use of time-resolved fluorescence, localisation of the protein within cells or model systems can be monitored. These fluorescence lifetime images (FLIM) have become the preferred technique for elucidating protein dynamics due to the fact that the fluorescence lifetime is an absolute measure, in the main independent of fluorophore concentration and intensity fluctuations caused by factors such as photobleaching. In this work we demonstrate the use of a time-resolved fluorescence microscopy, employing a high repetition rate laser excitation source applied to study the influence of a metal surface on fluorescence tagged protein and to elucidate viscosity using the fluorescence lifetime probe DASPMI. These were studied in a cellular environment (yeast) and in a model system based on a sol-gel derived material, in which silver nanostructures were formed in situ using irradiation from a semiconductor laser in CW mode incorporated on a compact time-resolved fluorescence microscope (HORIBA Scientific DeltaDiode and DynaMyc).

  15. Luminescent detection of hydrazine and hydrazine derivatives

    DOEpatents

    Swager, Timothy M [Newton, MA; Thomas, III, Samuel W.

    2012-04-17

    The present invention generally relates to methods for modulating the optical properties of a luminescent polymer via interaction with a species (e.g., an analyte). In some cases, the present invention provides methods for determination of an analyte by monitoring a change in an optical signal of a luminescent polymer upon exposure to an analyte. Methods of the present invention may be useful for the vapor phase detection of analytes such as explosives and toxins. The present invention also provides methods for increasing the luminescence intensity of a polymer, such as a polymer that has been photobleached, by exposing the luminescent polymer to a species such as a reducing agent.

  16. [Effect of procaine and procaine metabolites on coenzyme A and acetyl coenzyme A concentration in various tissues of the rat].

    PubMed

    Kietzmann, M; Kaemmerer, K

    1989-01-01

    In rats treated with procaine hydrochloride, diethylaminoethanol, monoethylaminoethanol, ethanolamine, as well as a combination of procaine hydrochloride and haematoporphyrine the ratio of acetyl coenzyme A and coenzyme A clearly was enhanced in the liver and to a minor extent in the cerebellum. In the tissue of cerebral cortex, heart, muscle and duodenum no corresponding effects were demonstrated. These findings, showing a further intermediary effect of orally administered procaine, can be interpreted as an influence in intermediary energy utilisation.

  17. Fluorescence recovery after photobleaching measured by confocal microscopy as a tool for the analysis of vesicular lipid transport and plasma membrane mobility

    NASA Astrophysics Data System (ADS)

    Schmitz, Gerd; Goetz, Alexandra; Orso, Evelyn; Rothe, Gregor

    1998-04-01

    The vesicular transport of lipids from the endoplasmic reticulum via the Golgi apparatus affects the composition of the plasma membrane. The purpose of our study was to develop an in vitro test system for characterization of vesicular lipid transport kinetics by using confocal microscopy and fluorescence recovery after photobleaching (FRAP). Fibroblasts from two patients homozygous for the hypercatabolic HDL deficiency syndrome Tangier disease and 4 control subjects were pulsed with the C6-NBD-ceramide for 30 minutes. Chase incubation at room temperature resulted in the metabolic accumulation of fluorescent C6-NBD-sphingolyelin and C6-NBD-glycosylceramides in the medial- and trans-Golgi region. Cells were analyzed with an inverted Leica TCS microscope. Calibration was performed through the analysis of diffusion of 50 nm microparticles embedded in media of different viscosity. An acousto optical tunable filter (AOTF) was used for the selective bleaching of the medial- and trans- Golgi region followed by analysis of the fluorescence recovery for 4 minutes. Post-bleach fluorescence recovery through the trans-Golgi-oriented transport of NBD-sphingomyelin was calculated from 2-dimensional scans. Tangier fibroblasts displayed a retarded recovery of fluorescence in the trans- Golgi region. This suggests that the vesicular transport of sphingomyelin and cholesterol is disturbed in Tangier disease confirming data from our laboratory generated with radiometabolites on whole cells. Our data suggest that FRAP analysis allows a sensitive kinetic and spatially resolved analysis of disturbances of vesicular lipid transport.

  18. Fluorescence recovery after photobleaching reveals regulation and distribution of connexin36 gap junction coupling within mouse islets of Langerhans

    PubMed Central

    Farnsworth, Nikki L; Hemmati, Alireza; Pozzoli, Marina; Benninger, Richard K P

    2014-01-01

    The pancreatic islets are central to the maintenance of glucose homeostasis through insulin secretion. Glucose-stimulated insulin secretion is tightly linked to electrical activity in β cells within the islet. Gap junctions, composed of connexin36 (Cx36), form intercellular channels between β cells, synchronizing electrical activity and insulin secretion. Loss of gap junction coupling leads to altered insulin secretion dynamics and disrupted glucose homeostasis. Gap junction coupling is known to be disrupted in mouse models of pre-diabetes. Although approaches to measure gap junction coupling have been devised, they either lack cell specificity, suitable quantification of coupling or spatial resolution, or are invasive. The purpose of this study was to develop fluorescence recovery after photobleaching (FRAP) as a technique to accurately and robustly measure gap junction coupling in the islet. The cationic dye Rhodamine 123 was used with FRAP to quantify dye diffusion between islet β cells as a measure of Cx36 gap junction coupling. Measurements in islets with reduced Cx36 verified the accuracy of this technique in distinguishing between distinct levels of gap junction coupling. Analysis of individual cells revealed that the distribution of coupling across the islet is highly heterogeneous. Analysis of several modulators of gap junction coupling revealed glucose- and cAMP-dependent modulation of gap junction coupling in islets. Finally, FRAP was used to determine cell population specific coupling, where no functional gap junction coupling was observed between α cells and β cells in the islet. The results of this study show FRAP to be a robust technique which provides the cellular resolution to quantify the distribution and regulation of Cx36 gap junction coupling in specific cell populations within the islet. Future studies utilizing this technique may elucidate the role of gap junction coupling in the progression of diabetes and identify mechanisms of gap

  19. Fluorescence recovery after photobleaching reveals regulation and distribution of connexin36 gap junction coupling within mouse islets of Langerhans.

    PubMed

    Farnsworth, Nikki L; Hemmati, Alireza; Pozzoli, Marina; Benninger, Richard K P

    2014-10-15

    The pancreatic islets are central to the maintenance of glucose homeostasis through insulin secretion. Glucose‐stimulated insulin secretion is tightly linked to electrical activity in β cells within the islet. Gap junctions, composed of connexin36 (Cx36), form intercellular channels between β cells, synchronizing electrical activity and insulin secretion. Loss of gap junction coupling leads to altered insulin secretion dynamics and disrupted glucose homeostasis. Gap junction coupling is known to be disrupted in mouse models of pre‐diabetes. Although approaches to measure gap junction coupling have been devised, they either lack cell specificity, suitable quantification of coupling or spatial resolution, or are invasive. The purpose of this study was to develop fluorescence recovery after photobleaching (FRAP) as a technique to accurately and robustly measure gap junction coupling in the islet. The cationic dye Rhodamine 123 was used with FRAP to quantify dye diffusion between islet β cells as a measure of Cx36 gap junction coupling. Measurements in islets with reduced Cx36 verified the accuracy of this technique in distinguishing between distinct levels of gap junction coupling. Analysis of individual cells revealed that the distribution of coupling across the islet is highly heterogeneous. Analysis of several modulators of gap junction coupling revealed glucose‐ and cAMP‐dependent modulation of gap junction coupling in islets. Finally, FRAP was used to determine cell population specific coupling, where no functional gap junction coupling was observed between α cells and β cells in the islet. The results of this study show FRAP to be a robust technique which provides the cellular resolution to quantify the distribution and regulation of Cx36 gap junction coupling in specific cell populations within the islet. Future studies utilizing this technique may elucidate the role of gap junction coupling in the progression of diabetes and identify mechanisms of

  20. Autumn photoproduction of carbon monoxide in Jiaozhou Bay, China

    NASA Astrophysics Data System (ADS)

    Ren, Chunyan; Yang, Guipeng; Lu, Xiaolan

    2014-06-01

    Carbon monoxide (CO) plays a significant role in global warming and atmospheric chemistry. Global oceans are net natural sources of atmospheric CO. CO at surface ocean is primarily produced from the photochemical degradation of chromophoric dissolved organic matter (CDOM). In this study, the effects of photobleaching, temperature and the origin (terrestrial or marine) of CDOM on the apparent quantum yields (AQY) of CO were studied for seawater samples collected from Jiaozhou Bay. Our results demonstrat that photobleaching, temperature and the origin of CDOM strongly affected the efficiency of CO photoproduction. The concentration, absorbance and fluorescence of CDOM exponentially decreased with increasing light dose. Terrestrial riverine organic matter could be more prone to photodegradation than the marine algae-derived one. The relationships between CO AQY and the dissolved organic carbon-specific absorption coefficient at 254 nm for the photobleaching study were nonlinear, whereas those of the original samples were strongly linear. This suggests that: 1) terrestrial riverine CDOM was more efficient than marine algae-derived CDOM for CO photoproduction; 2) aromatic and olefinic moieties of the CDOM pool were affected more strongly by degradation processes than by aliphatic ones. Water temperature and the origin of CDOM strongly affected the efficiency of CO photoproduction. The photoproduction rate of CO in autumn was estimated to be 31.98 μmol m-2 d-1 and the total DOC photomineralization was equivalent to 3.25%-6.35% of primary production in Jiaozhou Bay. Our results indicate that CO photochemistry in coastal areas is important for oceanic carbon cycle.

  1. Dual photon excitation microscopy and image threshold segmentation in live cell imaging during compression testing.

    PubMed

    Moo, Eng Kuan; Abusara, Ziad; Abu Osman, Noor Azuan; Pingguan-Murphy, Belinda; Herzog, Walter

    2013-08-09

    Morphological studies of live connective tissue cells are imperative to helping understand cellular responses to mechanical stimuli. However, photobleaching is a constant problem to accurate and reliable live cell fluorescent imaging, and various image thresholding methods have been adopted to account for photobleaching effects. Previous studies showed that dual photon excitation (DPE) techniques are superior over conventional one photon excitation (OPE) confocal techniques in minimizing photobleaching. In this study, we investigated the effects of photobleaching resulting from OPE and DPE on morphology of in situ articular cartilage chondrocytes across repeat laser exposures. Additionally, we compared the effectiveness of three commonly-used image thresholding methods in accounting for photobleaching effects, with and without tissue loading through compression. In general, photobleaching leads to an apparent volume reduction for subsequent image scans. Performing seven consecutive scans of chondrocytes in unloaded cartilage, we found that the apparent cell volume loss caused by DPE microscopy is much smaller than that observed using OPE microscopy. Applying scan-specific image thresholds did not prevent the photobleaching-induced volume loss, and volume reductions were non-uniform over the seven repeat scans. During cartilage loading through compression, cell fluorescence increased and, depending on the thresholding method used, led to different volume changes. Therefore, different conclusions on cell volume changes may be drawn during tissue compression, depending on the image thresholding methods used. In conclusion, our findings confirm that photobleaching directly affects cell morphology measurements, and that DPE causes less photobleaching artifacts than OPE for uncompressed cells. When cells are compressed during tissue loading, a complicated interplay between photobleaching effects and compression-induced fluorescence increase may lead to interpretations in

  2. Neutral Porphyrin Derivative Exerts Anticancer Activity by Targeting Cellular Topoisomerase I (Top1) and Promotes Apoptotic Cell Death without Stabilizing Top1-DNA Cleavage Complexes

    PubMed Central

    2017-01-01

    Camptothecin (CPT) selectively traps topoisomerase 1-DNA cleavable complexes (Top1cc) to promote anticancer activity. Here, we report the design and synthesis of a new class of neutral porphyrin derivative 5,10-bis(4-carboxyphenyl)-15, 20-bis(4-dimethylaminophenyl)porphyrin (compound 8) as a potent catalytic inhibitor of human Top1. In contrast to CPT, compound 8 reversibly binds with the free enzyme and inhibits the formation of Top1cc and promotes reversal of the preformed Top1cc with CPT. Compound 8 induced inhibition of Top1cc formation in live cells was substantiated by fluorescence recovery after photobleaching (FRAP) assays. We established that MCF7 cells treated with compound 8 trigger proteasome-mediated Top1 degradation, accumulate higher levels of reactive oxygen species (ROS), PARP1 cleavage, oxidative DNA fragmentation, and stimulate apoptotic cell death without stabilizing apoptotic Top1-DNA cleavage complexes. Finally, compound 8 shows anticancer activity by targeting cellular Top1 and preventing the enzyme from directly participating in the apoptotic process. PMID:29290109

  3. Investigating a continuous shear strain function for depth-dependent properties of native and tissue engineering cartilage using pixel-size data.

    PubMed

    Motavalli, Mostafa; Whitney, G Adam; Dennis, James E; Mansour, Joseph M

    2013-12-01

    A previously developed novel imaging technique for determining the depth dependent properties of cartilage in simple shear is implemented. Shear displacement is determined from images of deformed lines photobleached on a sample, and shear strain is obtained from the derivative of the displacement. We investigated the feasibility of an alternative systematic approach to numerical differentiation for computing the shear strain that is based on fitting a continuous function to the shear displacement. Three models for a continuous shear displacement function are evaluated: polynomials, cubic splines, and non-parametric locally weighted scatter plot curves. Four independent approaches are then applied to identify the best-fit model and the accuracy of the first derivative. One approach is based on the Akaiki Information Criteria, and the Bayesian Information Criteria. The second is based on a method developed to smooth and differentiate digitized data from human motion. The third method is based on photobleaching a predefined circular area with a specific radius. Finally, we integrate the shear strain and compare it with the total shear deflection of the sample measured experimentally. Results show that 6th and 7th order polynomials are the best models for the shear displacement and its first derivative. In addition, failure of tissue-engineered cartilage, consistent with previous results, demonstrates the qualitative value of this imaging approach. © 2013 Elsevier Ltd. All rights reserved.

  4. The sensitivity of normal brain and intracranially implanted VX2 tumour to interstitial photodynamic therapy.

    PubMed Central

    Lilge, L.; Olivo, M. C.; Schatz, S. W.; MaGuire, J. A.; Patterson, M. S.; Wilson, B. C.

    1996-01-01

    The applicability and limitations of a photodynamic threshold model, used to describe quantitatively the in vivo response of tissues to photodynamic therapy, are currently being investigated in a variety of normal and malignant tumour tissues. The model states that tissue necrosis occurs when the number of photons absorbed by the photosensitiser per unit tissue volume exceeds a threshold. New Zealand White rabbits were sensitised with porphyrin-based photosensitisers. Normal brain or intracranially implanted VX2 tumours were illuminated via an optical fibre placed into the tissue at craniotomy. The light fluence distribution in the tissue was measured by multiple interstitial optical fibre detectors. The tissue concentration of the photosensitiser was determined post mortem by absorption spectroscopy. The derived photodynamic threshold values for normal brain are significantly lower than for VX2 tumour for all photosensitisers examined. Neuronal damage is evident beyond the zone of frank necrosis. For Photofrin the threshold decreases with time delay between photosensitiser administration and light treatment. No significant difference in threshold is found between Photofrin and haematoporphyrin derivative. The threshold in normal brain (grey matter) is lowest for sensitisation by 5 delta-aminolaevulinic acid. The results confirm the very high sensitivity of normal brain to porphyrin photodynamic therapy and show the importance of in situ light fluence monitoring during photodynamic irradiation. Images Figure 1 Figure 4 Figure 5 Figure 6 Figure 7 PMID:8562339

  5. Revisiting point FRAP to quantitatively characterize anomalous diffusion in live cells.

    PubMed

    Daddysman, Matthew K; Fecko, Christopher J

    2013-02-07

    Fluorescence recovery after photobleaching (FRAP) is widely used to interrogate diffusion and binding of proteins in live cells. Herein, we apply two-photon excited FRAP with a diffraction limited bleaching and observation volume to study anomalous diffusion of unconjugated green fluorescence protein (GFP) in vitro and in cells. Experiments performed on dilute solutions of GFP reveal that reversible fluorophore bleaching can be mistakenly interpreted as anomalous diffusion. We derive a reaction-diffusion FRAP model that includes reversible photobleaching, and demonstrate that it properly accounts for these photophysics. We then apply this model to investigate the diffusion of GFP in HeLa cells and polytene cells of Drosophila larval salivary glands. GFP exhibits anomalous diffusion in the cytoplasm of both cell types and in HeLa nuclei. Polytene nuclei contain optically resolvable chromosomes, permitting FRAP experiments that focus separately on chromosomal or interchrosomal regions. We find that GFP exhibits anomalous diffusion in chromosomal regions but diffuses normally in regions devoid of chromatin. This observation indicates that obstructed transport through chromatin and not crowding by macromolecules is a source of anomalous diffusion in polytene nuclei. This behavior is likely true in other cells, so it will be important to account for this type of transport physics and for reversible photobleaching to properly interpret future FRAP experiments on DNA-binding proteins.

  6. The photokilling of bladder carcinoma cells in vitro by phenothiazine dyes.

    PubMed

    Fowler, G J; Rees, R C; Devonshire, R

    1990-09-01

    The potential photodynamic therapy photosensitizers Methylene Blue, Azure C, Methylene Violet, Thionine, Methylene Green, Haematoporphyrin, Nile Blue A, chloroaluminium phthalocyanine and bis-aluminium phthalocyanine were examined for their photoeffects and dark toxicity against a human superficial bladder carcinoma cell-line. By examination of [3H]thymidine uptake into dye-treated cells after irradiation with a copper-vapour pumped dye laser, it was found that Methylene Blue was the most phototoxic and dark toxic of all the dyes tested, suggesting that the dye might be of some use as a topically applied photodrug for use in photodynamic therapy of superficial or early-recurring carcinomas.

  7. The effect of surface irradiance on the absorption spectrum of chromophoric dissolved organic matter in the global ocean

    NASA Astrophysics Data System (ADS)

    Swan, Chantal M.; Nelson, Norman B.; Siegel, David A.; Kostadinov, Tihomir S.

    2012-05-01

    The cycling pathways of chromophoric dissolved organic matter (CDOM) within marine systems must be constrained to better assess the impact of CDOM on surface ocean photochemistry and remote sensing of ocean color. Photobleaching, the loss of absorption by CDOM due to light exposure, is the primary sink for marine CDOM. Herein the susceptibility of CDOM to photobleaching by sea surface-level solar radiation was examined in 15 samples collected from wide-ranging open ocean regimes. Samples from the Pacific, Atlantic, Indian and Southern Oceans were irradiated over several days with full-spectrum light under a solar simulator at in situ temperature in order to measure photobleaching rate and derive an empirical matrix, ɛsurf (m-1 μEin-1), which quantifies the effect of surface irradiance on the spectral absorption of CDOM. Irradiation responses among the ocean samples were similar within the ultraviolet (UV) region of the spectrum spanning 300-360 nm, generally exhibiting a decrease in the CDOM absorption coefficient (m-1) and concomitant increase in the CDOM spectral slope parameter, S (nm-1). However, an unexpected irradiation-induced increase in CDOM absorption between approximately 360 and 500 nm was observed for samples from high-nutrient low-chlorophyll (HNLC) environments. This finding was linked to the presence of dissolved nitrate and may explain discrepancies in action spectra for dimethylsulfide (DMS) photobleaching observed between the Equatorial Pacific and Subtropical North Atlantic Oceans. The nitrate-to-phosphate ratio explained 27-70% of observed variability in ɛsurf at observation wavelengths of 330-440 nm, while the initial spectral slope of the samples explained up to 52% of variability in ɛsurf at observation wavelengths of 310-330 nm. These results suggest that the biogeochemical and solar exposure history of the water column, each of which influence the chemical character and thus the spectral quality of CDOM and its photoreactivity, are the

  8. Spatiotemporal Distribution, Sources, and Photobleaching Imprint of Dissolved Organic Matter in the Yangtze Estuary and Its Adjacent Sea Using Fluorescence and Parallel Factor Analysis

    PubMed Central

    Li, Penghui; Chen, Ling; Zhang, Wen; Huang, Qinghui

    2015-01-01

    To investigate the seasonal and interannual dynamics of dissolved organic matter (DOM) in the Yangtze Estuary, surface and bottom water samples in the Yangtze Estuary and its adjacent sea were collected and characterized using fluorescence excitation-emission matrices (EEMs) and parallel factor analysis (PARAFAC) in both dry and wet seasons in 2012 and 2013. Two protein-like components and three humic-like components were identified. Three humic-like components decreased linearly with increasing salinity (r>0.90, p<0.001), suggesting their distribution could primarily be controlled by physical mixing. By contrast, two protein-like components fell below the theoretical mixing line, largely due to microbial degradation and removal during mixing. Higher concentrations of humic-like components found in 2012 could be attributed to higher freshwater discharge relative to 2013. There was a lack of systematic patterns for three humic-like components between seasons and years, probably due to variations of other factors such as sources and characteristics. Highest concentrations of fluorescent components, observed in estuarine turbidity maximum (ETM) region, could be attributed to sediment resuspension and subsequent release of DOM, supported by higher concentrations of fluorescent components in bottom water than in surface water at two stations where sediments probably resuspended. Meanwhile, photobleaching could be reflected from the changes in the ratios between fluorescence intensity (Fmax) of humic-like components and chromophoric DOM (CDOM) absorption coefficient (a355) along the salinity gradient. This study demonstrates the abundance and composition of DOM in estuaries are controlled not only by hydrological conditions, but also by its sources, characteristics and related estuarine biogeochemical processes. PMID:26107640

  9. Uninterrupted monitoring of drug effects in human-induced pluripotent stem cell-derived cardiomyocytes with bioluminescence Ca2+ microscopy.

    PubMed

    Suzuki, Kazushi; Onishi, Takahito; Nakada, Chieko; Takei, Shunsuke; Daniels, Matthew J; Nakano, Masahiro; Matsuda, Tomoki; Nagai, Takeharu

    2018-05-18

    Cardiomyocytes derived from human-induced pluripotent stem cells are a powerful platform for high-throughput drug screening in vitro. However, current modalities for drug testing, such as electrophysiology and fluorescence imaging have inherent drawbacks. To circumvent these problems, we report the development of a bioluminescent Ca 2+ indicator GmNL(Ca 2+ ), and its application in a customized microscope for high-throughput drug screening. GmNL(Ca 2+ ) gives a 140% signal change with Ca 2+ , and can image drug-induced changes of Ca 2+ dynamics in cultured cells. Since bioluminescence requires application of a chemical substrate, which is consumed over ~ 30 min we made a dedicated microscope with automated drug dispensing inside a light-tight box, to control drug addition. To overcome thermal instability of the luminescent substrate, or small molecule, dual climate control enables distinct temperature settings in the drug reservoir and the biological sample. By combining GmNL(Ca 2+ ) with this adaptation, we could image spontaneous Ca 2+ transients in cultured cardiomyocytes and phenotype their response to well-known drugs without accessing the sample directly. In addition, the bioluminescent strategy demonstrates minimal perturbation of contractile parameters and long-term observation attributable to lack of phototoxicity and photobleaching. Overall, bioluminescence may enable more accurate drug screening in a high-throughput manner.

  10. Influence of photo- and thermal bleaching on pre-irradiation low water peak single mode fibers

    NASA Astrophysics Data System (ADS)

    Yin, Jianchong; Wen, Jianxiang; Luo, Wenyun; Xiao, Zhongyin; Chen, Zhenyi; Wang, Tingyun

    2011-12-01

    Reducing the radiation-induced transmission loss in low water peak single mode fiber (LWP SMF) has been investigated by using photo-bleaching method with 980nm pump light source and using thermal-bleaching method with temperature control system. The results show that the radiation-induced loss of pre-irradiation optical fiber can be reduced effectively with the help of photo-bleaching or thermal-bleaching. Although the effort of photo-bleaching is not as significant as thermal-bleaching, by using photo-bleaching method, the loss of fiber caused by radiation-induced defects can be reduced best up to 49% at 1310nm and 28% at 1550nm in low pre-irradiation condition, the coating of the fiber are not destroyed, and the rehabilitating time is just several hours, while self-annealing usually costs months' time. What's more, the typical high power LASER for photo-bleaching can be 980nm pump Laser Diode, which is very accessible.

  11. Cost-effective elimination of lipofuscin fluorescence from formalin-fixed brain tissue by white phosphor light emitting diode array.

    PubMed

    Sun, Yulong; Chakrabartty, Avi

    2016-12-01

    Autofluorescence of aldehyde-fixed tissues greatly hinders fluorescence microscopy. In particular, lipofuscin, an autofluorescent component of aged brain tissue, complicates fluorescence imaging of tissue in neurodegenerative diseases. Background and lipofuscin fluorescence can be reduced by greater than 90% through photobleaching using white phosphor light emitting diode arrays prior to treatment with fluorescent probes. We compared the effect of photobleaching versus established chemical quenchers on the quality of fluorescent staining in formalin-fixed brain tissue of frontotemporal dementia with tau-positive inclusions. Unlike chemical quenchers, which reduced fluorescent probe signals as well as background, photobleaching treatment had no effect on probe fluorescence intensity while it effectively reduced background and lipofuscin fluorescence. The advantages and versatility of photobleaching over established methods are discussed.

  12. Simple Elimination of Background Fluorescence in Formalin-Fixed Human Brain Tissue for Immunofluorescence Microscopy.

    PubMed

    Sun, Yulong; Ip, Philbert; Chakrabartty, Avijit

    2017-09-03

    Immunofluorescence is a common method used to visualize subcellular compartments and to determine the localization of specific proteins within a tissue sample. A great hindrance to the acquisition of high quality immunofluorescence images is endogenous autofluorescence of the tissue caused by aging pigments such as lipofuscin or by common sample preparation processes such as aldehyde fixation. This protocol describes how background fluorescence can be greatly reduced through photobleaching using white phosphor light emitting diode (LED) arrays prior to treatment with fluorescent probes. The broad-spectrum emission of white phosphor LEDs allow for bleaching of fluorophores across a range of emission peaks. The photobleaching apparatus can be constructed from off-the-shelf components at very low cost and offers an accessible alternative to commercially available chemical quenchers. A photobleaching pre-treatment of the tissue followed by conventional immunofluorescence staining generates images free of background autofluorescence. Compared to established chemical quenchers which reduced probe as well as background signals, photobleaching treatment had no effect on probe fluorescence intensity while it effectively reduced background and lipofuscin fluorescence. Although photobleaching requires more time for pre-treatment, higher intensity LED arrays may be used to reduce photobleaching time. This simple method can potentially be applied to a variety of tissues, particularly postmitotic tissues that accumulate lipofuscin such as the brain and cardiac or skeletal muscles.

  13. Lethal photosensitization of Helicobacter species

    NASA Astrophysics Data System (ADS)

    Millson, Charles E.; Wilson, Michael; MacRobert, Alexander J.; Thurrell, Wendy; Mlkvy, Peter; Davies, Claire; Bown, Stephen G.

    1995-01-01

    Helicobacter pylori (H. pylori) is associated with a large number of gastroduodenal disorders. Clearance of the bacteria has been shown to benefit patients with duodenal ulcers, gastric ulcers, and certain rare types of gastric tumors. Broad-spectrum antibiotics are the mainstay of current treatment strategies but side-effects, poor compliance, and drug resistance limit their usefulness. We sensitized H. pylori with toluidine blue, haematoporphyrin derivative, aluminum disulphonated phthalocyanine, methylene blue or protoporphyrin IX prior to exposure to low-power laser light from either a gallium aluminum arsenide laser or a helium neon gas laser. All 5 sensitizers caused reductions of greater than 1000-fold in the number of viable bacteria. Light alone had no effect and only HpD caused a significant decrease in bacterial numbers without laser light. Next, we sensitized H. mustelae on explanted ferret gastric mucosa (ex vivo) with the same sensitizers and exposed them to light from a copper vapor pumped dye laser tuned appropriately. MB caused significant reductions in bacterial counts. Successful lethal photosensitization of Helicobacter pylori both in vitro and ex vivo raises the possibility of a local method for eradicating the bacteria, especially as the bacteria are only found in those parts of the upper gastrointestinal tract that are accessible to the endoscope.

  14. THE EFFECT OF PHOTOPIGMENT BLEACHING ON FUNDUS AUTOFLUORESCENCE IN ACUTE CENTRAL SEROUS CHORIORETINOPATHY.

    PubMed

    Choi, Kwang-Eon; Yun, Cheolmin; Kim, Young-Ho; Kim, Seong-Woo; Oh, Jaeryung; Huh, Kuhl

    2017-03-01

    To evaluate the effect of photobleaching on fundus autofluorescence (FAF) images in acute central serous chorioretinopathy. We obtained prephotobleaching and postphotobleaching images using an Optomap 200Tx, and photobleaching was induced with a Heidelberg Retina Angiograph 2. Degrees of photobleaching were assessed as grayscale values in Optomap images. Concordances among the three kinds of images were analyzed. Hyper-AF lesions in prephotobleaching images were classified as Type 1 (changed to normal-AF after photobleaching) and Type 2 (unchanged after photobleaching). The FAF composite patterns of central serous chorioretinopathy lesions were classified as diffuse or mottled. Initial and final best-corrected visual acuity, central retinal thickness, and disease duration were compared according to fovea FAF type. Forty-one eyes of 41 patients were analyzed. The lesion brightness of postphotobleaching Optomap FAF showed greater concordance with Heidelberg Retina Angiograph 2 FAF (94.74%) than the prephotobleaching Optomap FAF (80.49%). Eyes with Type 1 fovea had greater initial and final best-corrected visual acuity (20/23 vs. 20/41, 20/21 vs. 20/32, P < 0.0001, P = 0.001, respectively) and shorter disease duration (19.68 ± 12.98 vs. 51.55 ± 44.98 days, P = 0.043) than those with Type 2 fovea. However, eyes with diffuse Type 2 fovea had only lower initial and final best-corrected visual acuity (20/23 vs. 20/45, 20/21 vs. 20/36, P < 0.0001, P < 0.0001, respectively) than those with Type 1 fovea. Understanding the photobleaching effect is necessary for the accurate interpretation of FAF images. Furthermore, comparing prephotobleaching and postphotobleaching FAF images may be helpful for estimation of lesion status in central serous chorioretinopathy.

  15. Transposase-Derived Proteins FHY3/FAR1 Interact with PHYTOCHROME-INTERACTING FACTOR1 to Regulate Chlorophyll Biosynthesis by Modulating HEMB1 during Deetiolation in Arabidopsis[W

    PubMed Central

    Tang, Weijiang; Wang, Wanqing; Chen, Dongqin; Ji, Qiang; Jing, Yanjun; Wang, Haiyang; Lin, Rongcheng

    2012-01-01

    Successful chlorophyll biosynthesis during initial light exposure is critical for plant survival and growth, as excess accumulation of chlorophyll precursors in darkness can cause photooxidative damage to cells. Therefore, efficient mechanisms have evolved to precisely regulate chlorophyll biosynthesis in plants. Here, we identify FAR-RED ELONGATED HYPOCOTYL3 (FHY3) and FAR-RED IMPAIRED RESPONSE1 (FAR1), two transposase-derived transcription factors, as positive regulators of chlorophyll biosynthesis in Arabidopsis thaliana. We show that null mutations in FHY3 and FAR1 cause reduced protochlorophyllide (a precursor of chlorophyll) levels in darkness and less photobleaching in the light. We find that FHY3 directly binds to the promoter and activates expression of HEMB1, which encodes 5-aminolevulinic acid dehydratase in the chlorophyll biosynthetic pathway. We reveal that PHYTOCHROME-INTERACTING FACTOR1 physically interacts with the DNA binding domain of FHY3, thereby partly repressing FHY3/FAR1-activated HEMB1 expression. Strikingly, FHY3 expression is upregulated by white light. In addition, our genetic data indicate that overexpression, severe reduction, or lack of HEMB1 impairs plant growth and development. Together, our findings reveal a crucial role of FHY3/FAR1 in regulating chlorophyll biosynthesis, thus uncovering a new layer of regulation by which light promotes plant dark–light transition in early seedling development. PMID:22634759

  16. Integration of Fluorescence Differential Path-Length Spectroscopy to Photodynamic Therapy of the Head and Neck Tumors is Useful in Monitoring Clinical Outcome

    NASA Astrophysics Data System (ADS)

    Karakullukcu, Baris; Kanick, Stephen; Aans, Jan Bonne; Sterenborg, Henricus; Tan, Bing; Amelink, Arjen; Robinson, Dominic

    2015-04-01

    The use of fluorescence differential pathlength spectroscopy (FDPS) has the potential to provide real-time information on photosensitiser pharmacokinetics, vascular physiology and photosensitizer photobleaching based dosimetry of tumors in the oral cavity receiving m-tetrahydroxyphenylchlorin (mTHPC) photodynamic therapy (PDT). Reflectance spectra can be used provide quantitative values of oxygen saturation, blood volume fraction, blood vessel diameter, and to determine the local optical properties that can be used to correct raw fluorescence for tissue absorption. Patients and methods: Twenty-seven lesions in the oral cavity, either dysplasias or cancer were interrogated using FDPS, before and immediately after the therapeutic illumination. The average tumor center to normal mucosa ratio of fluorescence was 1.50 ± 0.66. mTHPC photobleaching was observed in 24 of the lesions treated. The average extent of photobleaching was 81% ± 17%. Information from FDPS spectroscopy coupled with the clinical results of the treatment identified 3 types of correctable errors in the application of mTHPC-PDT: Two patients exhibited very low concentrations of photosensitizer in tumour center, indicating an ineffective i.v. injection of photosensitiser or an erroneous systemic distribution of mTHPC. In one in tumor we observed no photobleaching accompanied by a high blood volume fraction in the illuminated tissue, suggesting that the presence of blood prevented therapeutic light reaching the target tissue. All 3 of the these lesions had no clinical response to PDT. In four patients we observed less than 50% photobleaching at the tumor margins , suggesting a possible geographic miss. One patient in this group had a recurrence within 2 months after PDT even though the initial response was good. The integration of FDPS to clinical PDT yields data on tissue physiology, photosensitiser content and photobleaching that can help identify treatment errors that can potentially be corrected.

  17. Cysteine optical sensing with an up-conversion host and two chemosensors derived from rhodamine: Construction, characterization and performance

    NASA Astrophysics Data System (ADS)

    Lin, Chen; Zhigang, Fang

    2017-03-01

    This paper focused on two rhodamine chemosensors for cysteine optical sensing. To minimize their photobleaching caused by excitation light, up-conversion NaYF4:Yb3 +/Er3 + nanocrystals were prepared and used as excitation host. Photophysical measurement on this host and the two chemosensors suggested that chemosensor absorption matched well with host emission. An efficient energy transfer between them was discussed and confirmed by their spectral analysis and emission lifetime comparison. Job's plot suggested that our chemosensors followed a simple recognition mechanism towards cysteine with binding stoichiometry of 1:1. Both chemosensors showed emission "off-on" effect triggered by cysteine and good photostability. Linear working curves with maximum sensitivity of 2.61 were obtained. S substituent was positive to improve selectivity.

  18. Role of photoacoustic effects occurring at laser perforation of skin and laser transcutaneous drug delivery

    NASA Astrophysics Data System (ADS)

    Zharov, Vladimir P.; Latyshev, Alexei S.

    2000-05-01

    A further study is given of photoacoustic (PA) drug delivery technologies using an Er:YAG laser, with the main emphasis being placed on the laser perforation of skin and on PA impregnation using an additional covering quartz plate. A mathematical model based on Fick's law for PA impregnation with regard to a free and rigid interface is considered. The histological examination of the perforated guinea-pig skin ex vivo shoed that a powerful PA wave forced skin epidermis to be bent inwards. In biopsies taken 15 min later the nuclei pyknosis of epidermis cells lining the perforated channel was observed. In biopsies obtained 36 hours later an insignificant necrotic lesion remained, whereas 120 hours laser the cells recovered. I twas shown experimentally that the PA signal increased 30 times after applying a quartz plate over the drug solution, which substantially enhanced drug penetration through the skin .The dependancies were obtained of the penetration depth of the haematoporphyrin derivative photosensitizer versus the number of laser pulses and the pressing force applied to the quartz plate. The chromatographic fractionation of Diprospan and Dexamethasone hormonal preparations prior to and after the action of 200 Er:YAG laser-induced PA waves demonstrated that no additional chemical agents resulting from drug dissociation were detected. The application of laser drug delivery methods in respect of treating dermatological diseases is also discussed.

  19. Imaging protein complex formation in the autophagy pathway: analysis of the interaction of LC3 and Atg4BC74A in live cells using Förster resonance energy transfer and fluorescence recovery after photobleaching

    NASA Astrophysics Data System (ADS)

    Kraft, Lewis J.; Kenworthy, Anne K.

    2012-01-01

    The protein microtubule-associated protein 1, light chain 3 (LC3) functions in autophagosome formation and plays a central role in the autophagy pathway. Previously, we found LC3 diffuses more slowly in cells than is expected for a freely diffusing monomer, suggesting it may constitutively associate with a macromolecular complex containing other protein components of the pathway. In the current study, we used Förster resonance energy transfer (FRET) microscopy and fluorescence recovery after photobleaching (FRAP) to investigate the interactions of LC3 with Atg4BC74A, a catalytically inactive mutant of the cysteine protease involved in lipidation and de-lipidation of LC3, as a model system to probe protein complex formation in the autophagy pathway. We show Atg4BC74A is in FRET proximity with LC3 in both the cytoplasm and nucleus of living cells, consistent with previous biochemical evidence that suggests these proteins directly interact. In addition, overexpressed Atg4BC74A diffuses significantly more slowly than predicted based on its molecular weight, and its translational diffusion coefficient is significantly slowed upon coexpression with LC3 to match that of LC3 itself. Taken together, these results suggest Atg4BC74A and LC3 are contained within the same multiprotein complex and that this complex exists in both the cytoplasm and nucleoplasm of living cells.

  20. Enhanced labeling density and whole-cell 3D dSTORM imaging by repetitive labeling of target proteins.

    PubMed

    Venkataramani, Varun; Kardorff, Markus; Herrmannsdörfer, Frank; Wieneke, Ralph; Klein, Alina; Tampé, Robert; Heilemann, Mike; Kuner, Thomas

    2018-04-03

    With continuing advances in the resolving power of super-resolution microscopy, the inefficient labeling of proteins with suitable fluorophores becomes a limiting factor. For example, the low labeling density achieved with antibodies or small molecule tags limits attempts to reveal local protein nano-architecture of cellular compartments. On the other hand, high laser intensities cause photobleaching within and nearby an imaged region, thereby further reducing labeling density and impairing multi-plane whole-cell 3D super-resolution imaging. Here, we show that both labeling density and photobleaching can be addressed by repetitive application of trisNTA-fluorophore conjugates reversibly binding to a histidine-tagged protein by a novel approach called single-epitope repetitive imaging (SERI). For single-plane super-resolution microscopy, we demonstrate that, after multiple rounds of labeling and imaging, the signal density is increased. Using the same approach of repetitive imaging, washing and re-labeling, we demonstrate whole-cell 3D super-resolution imaging compensated for photobleaching above or below the imaging plane. This proof-of-principle study demonstrates that repetitive labeling of histidine-tagged proteins provides a versatile solution to break the 'labeling barrier' and to bypass photobleaching in multi-plane, whole-cell 3D experiments.

  1. Fluorescence Guided PDT for Optimization of Skin Cancer Treatment

    NASA Astrophysics Data System (ADS)

    Blanco, Kate; Moriyama, Lilian; Inada, Natalia; Kurachi, Cristina; Salvio, Ana; Leite, Everson; Menezes, Priscila; Bagnato, Vanderlei

    2015-04-01

    The photodynamic therapy (PDT) is an alternative technique that can be indicated for superficial basal cell carcinoma (sBCC), Bowen’s disease and actinic keratosis with high efficiency. The objective of this study is to present the importance of fluorescence imaging for PDT guidance and monitoring in real time. Confirming that the lesion is well prepared and the photosensitizer shows a homogenous distribution, the outcome after few PDT sessions will be positive and the recurrence should be lower. Our proposition in this study is use the widefield fluorescence imaging to evaluate the PDT protocol in situ and in real time for each lesion. This evaluation procedure is performed in two steps: first with the monitoring of the production of protoporphyrin IX (PpIX) induced by methyl aminolevulinate (MAL), an derivative of 5-aminolevulinic acid (ALA) and second with the detection of PpIX photobleaching after illumination. The fluorescence images provide information correlated with distinct clinical features and with the treatment outcome. Eight BCC lesions are presented and discussed in this study. Different fluorescence patterns of PpIX production and photobleaching could be correlated with the treatment response. The presented results show the potential of using widefield fluorescence imaging as a guidance tool to customized PDT.

  2. Robust label-free biosensing using microdisk laser arrays with on-chip references.

    PubMed

    Wondimu, S F; Hippler, M; Hussal, C; Hofmann, A; Krämmer, S; Lahann, J; Kalt, H; Freude, W; Koos, C

    2018-02-05

    Whispering-gallery mode (WGM) microdisk lasers show great potential for highly sensitive label-free detection in large-scale sensor arrays. However, when used in practical applications under normal ambient conditions, these devices suffer from temperature fluctuations and photobleaching. Here we demonstrate that these challenges can be overcome by a novel referencing scheme that allows for simultaneous compensation of temperature drift and photobleaching. The technique relies on reference structures protected by locally dispensed passivation materials, and can be scaled to extended arrays of hundreds of devices. We prove the viability of the concept in a series of experiments, demonstrating robust and sensitive label-free detection over a wide range of constant or continuously varying temperatures. To the best of our knowledge, these measurements represent the first demonstration of biosensing in active WGM devices with simultaneous compensation of both photobleaching and temperature drift.

  3. Ex-vivo treatment of gastric Helicobacter infection by photodynamic therapy.

    PubMed

    Millson, C E; Wilson, M; MacRobert, A J; Bown, S G

    1996-01-01

    Attempts to develop PDT for eradication of Helicobacter infection have only been successful in vitro. We have investigated the effect of topical sensitization (except ALA) of Helicobacter mustelae on explanted ferret gastric mucosa using one of five sensitizers (methylene blue (MB), toluidine blue O (TBO), phthalocyanine, haematoporphyrin derivative and 5-aminolavulinic acid), followed by irradiation with an appropriately tuned copper vapour pumped dye laser. A 90% reduction in counts of bacteria sensitized with 0.75 mg TBO kg-1 were seen after irradiation with 200 J cm-2. Concentrations of MB of 0.75 mg kg-1 and 7.5 mg kg-1 were not toxic to H. mustelae, but the further addition of 20 J cm-2 laser light reduced colony counts by more than 99%. MB at a concentration of 75 mg kg-1 exhibited significant dark toxicity towards H. mustelae, but further addition of 20 J cm-2 laser light resulted in near eradication of all colonies. The remaining three compounds were ineffective. Finally, we studied the microscopic fluorescence distribution of MB (7.5 mg kg-1) on ferret gastric mucosa after topical administration. Fluorescence was greatest in the superficial mucosal layer, upon which lies the bacteria. However, from experiments on rats, the energy required to kill the sensitized bacteria was insufficient to damage the underlying mucosa. We conclude that Helicobacter can be killed on host mucosal epithelium following topical administration of MB and subsequent exposure to laser light.

  4. Topical photosan-mediated photodynamic therapy for DMBA-induced hamster buccal pouch early cancer lesions: an in vivo study

    NASA Astrophysics Data System (ADS)

    Hsu, Yih-Chih; Chang, Walter Hong-Shong; Chang, Junn-Liang; Liu, Kuang-Ting; Chiang, Chun-Pin; Liu, Chung-Ji; Chen, Chih-Ping

    2011-03-01

    Oral cancer has becomes the most prominent cancer disease in recent years in Taiwan. The reason is the betel nut chewing habit combing with smoking and alcohol-drinking lifestyle of people results in oral cancer becomes the fastest growth incident cancer amongst other major cancer diseases. In previous studies showed that photosan, haematoporphyrin derivative (HPD), has demonstrated effective PDT results on human head and neck disease studies. To avoid the systemic phototoxic effect of photosan, this study was designed to use a topical photosan-mediated PDT for treatment of DMBA-induced hamster buccal pouch cancerous lesions. DMBA was applied to one of the buccal pouches of hamsters thrice a week for 10 to 12 weeks. Cancerous lesions were induced and proven by histological examination. These DMBA-induced cancerous lesions were used for testing the efficacy of topical photosan-mediated PDT. Before PDT, fluorescence spectroscopy was used to determine when photosan reached its peak level in the lesional epithelial cells after topical application of photosan gel. We found that photosan reached its peak level in cancerous lesions about 13.5 min after topical application of photosan gel. The cancerous lesions in hamsters were then treated with topical photosan-mediated PDT (fluence rate: 600 mW/cm2; light exposure dose 200 J/cm2) using the portable Lumacare 635 nm fiber-guided light device. Visual examination demonstrated that topical photosan-mediated PDT was an applicable treatment modality for DMBA-induced hamster buccal pouch cancerous lesions.

  5. Experts' understanding of partial derivatives using the partial derivative machine

    NASA Astrophysics Data System (ADS)

    Roundy, David; Weber, Eric; Dray, Tevian; Bajracharya, Rabindra R.; Dorko, Allison; Smith, Emily M.; Manogue, Corinne A.

    2015-12-01

    [This paper is part of the Focused Collection on Upper Division Physics Courses.] Partial derivatives are used in a variety of different ways within physics. Thermodynamics, in particular, uses partial derivatives in ways that students often find especially confusing. We are at the beginning of a study of the teaching of partial derivatives, with a goal of better aligning the teaching of multivariable calculus with the needs of students in STEM disciplines. In this paper, we report on an initial study of expert understanding of partial derivatives across three disciplines: physics, engineering, and mathematics. We report on the central research question of how disciplinary experts understand partial derivatives, and how their concept images of partial derivatives differ, with a focus on experimentally measured quantities. Using the partial derivative machine (PDM), we probed expert understanding of partial derivatives in an experimental context without a known functional form. In particular, we investigated which representations were cued by the experts' interactions with the PDM. Whereas the physicists and engineers were quick to use measurements to find a numeric approximation for a derivative, the mathematicians repeatedly returned to speculation as to the functional form; although they were comfortable drawing qualitative conclusions about the system from measurements, they were reluctant to approximate the derivative through measurement. On a theoretical front, we found ways in which existing frameworks for the concept of derivative could be expanded to include numerical approximation.

  6. DURIP - Upgrade of the Meridian ACAS-470 for Toxicological Research

    DTIC Science & Technology

    1990-01-18

    Flow Cytometry and Fluorescence Recovery After PhotobleachIng. Scanningi Microscopy. 2 14) 21 53 -2163. Li~j. Z-Y, YMEL Sanders. and V W Hu. (1989...riinn’I.~ - correlation with increasing tumorigenicity. Northern an- alysis showed reduced levels of connexin 43 in ce!1 lines Fpi~m~iIisFP - exhibiting...cells to the curcinogenic process", In: Mouse Liver CarcinoQgnesis; 3 Mechanisms and Species Comrarisons, T.J. Slaga, Ed., Alan R. Liss, Inc., New

  7. Continuous-wave organic dye lasers and methods

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

    Shapira, Ofer; Chua, Song-Liang; Zhen, Bo

    2014-09-16

    An organic dye laser produces a continuous-wave (cw) output without any moving parts (e.g., without using flowing dye streams or spinning discs of solid-state dye media to prevent photobleaching) and with a pump beam that is stationary with respect to the organic dye medium. The laser's resonant cavity, organic dye medium, and pump beam are configured to excite a lasing transition over a time scale longer than the associated decay lifetimes in the organic dye medium without photobleaching the organic dye medium. Because the organic dye medium does not photobleach when operating in this manner, it may be pumped continuouslymore » so as to emit a cw output beam. In some examples, operation in this manner lowers the lasing threshold (e.g., to only a few Watts per square centimeter), thereby facilitating electrical pumping for cw operation.« less

  8. Correlative FRET: new method improves rigor and reproducibility in determining distances within synaptic nanoscale architecture

    NASA Astrophysics Data System (ADS)

    Shinogle-Decker, Heather; Martinez-Rivera, Noraida; O'Brien, John; Powell, Richard D.; Joshi, Vishwas N.; Connell, Samuel; Rosa-Molinar, Eduardo

    2018-02-01

    A new correlative Förster Resonance Energy Transfer (FRET) microscopy method using FluoroNanogold™, a fluorescent immunoprobe with a covalently attached Nanogold® particle (1.4nm Au), overcomes resolution limitations in determining distances within synaptic nanoscale architecture. FRET by acceptor photobleaching has long been used as a method to increase fluorescence resolution. The transfer of energy from a donor to an acceptor generally occurs between 10-100Å, which is the relative distance between the donor molecule and the acceptor molecule. For the correlative FRET microscopy method using FluoroNanogold™, we immuno-labeled GFP-tagged-HeLa-expressing Connexin 35 (Cx35) with anti-GFP and with anti-Cx35/36 antibodies, and then photo-bleached the Cx before processing the sample for electron microscopic imaging. Preliminary studies reveal the use of Alexa Fluor® 594 FluoroNanogold™ slightly increases FRET distance to 70Å, in contrast to the 62.5Å using AlexaFluor 594®. Preliminary studies also show that using a FluoroNanogold™ probe inhibits photobleaching. After one photobleaching session, Alexa Fluor 594® fluorescence dropped to 19% of its original fluorescence; in contrast, after one photobleaching session, Alexa Fluor 594® FluoroNanogold™ fluorescence dropped to 53% of its original intensity. This result confirms that Alexa Fluor 594® FluoroNanogold™ is a much better donor probe than is Alexa Fluor 594®. The new method (a) creates a double confirmation method in determining structure and orientation of synaptic architecture, (b) allows development of a two-dimensional in vitro model to be used for precise testing of multiple parameters, and (c) increases throughput. Future work will include development of FluoroNanogold™ probes with different sizes of gold for additional correlative microscopy studies.

  9. Distribution and photoreactivity of chromophoric dissolved organic matter in northern Gulf of Mexico shelf waters

    NASA Astrophysics Data System (ADS)

    Shank, G. Christopher; Evans, Anne

    2011-07-01

    The distribution and photoreactivity of chromophoric dissolved organic matter (CDOM) in the northern Gulf of Mexico along the Louisiana coastal shelf were examined during three cruises in summer 2007, fall 2007, and summer 2008. The influence of the Mississippi River plume was clearly evident as CDOM levels (defined as a305) and dissolved organic carbon (DOC) concentrations were well-correlated with salinity during all cruises. Elevated CDOM and CDOM:DOC ratios of surface samples collected offshore of Atchafalaya Bay and the Breton-Chandeleur Sound complex indicated emanations of organic-rich waters from coastal wetlands are also an important source to nearshore shelf waters. Generally, CDOM and DOC levels were highest in surface waters and decreased with depth, but during summer 2007 and summer 2008, CDOM levels in near-bottom samples were occasionally higher than at mid-depths without concomitant increases in DOC. CDOM photobleaching was measured during 24 irradiations using a SunTest XLS+ solar simulator with photobleaching rate coefficients ( k305) ranging from 0.011 to 0.32 h -1. For fall 2007 and summer 2008, higher k305 values were generally observed in samples with higher initial CDOM levels. However, samples collected during summer 2007 did not exhibit a similar pattern nor were there differences in photobleaching rates between surface and bottom samples. Spectral slope coefficients ( S275-295 or S350-400) and DOC levels were largely unchanged after 24 h irradiations. Modeled CDOM photobleaching for northern Gulf of Mexico mid-shelf waters predicts that during the summer when solar irradiance is high and the water column becomes stratified, nearly 90% of the CDOM in the upper 1 m may be lost to photobleaching, with losses up to 20% possible even at 10 m depth.

  10. Improved Bilayer Resist System Using Contrast-Enhanced Lithography With Water-Soluble Photopolymer

    NASA Astrophysics Data System (ADS)

    Sasago, Masaru; Endo, Masayuki; Hirai, Yoshihiko; Ogawa, Kazufurni; Ishihara, Takeshi

    1986-07-01

    A new water-soluble contract enhanced material, WSP (Water-soluble Photopolymer), has been developed. The WSP is composed of a mainpolymer and a photobleachable reagents. The mainpolymer is a water-soluble polymer mixed with pullulan (refined through biotechnological process) and polyvinyl-pyrolidone (PVP). The photo-bleachable reagent is of a diazonium compound gorup. The introduction of the mainpolymer and photobleach-able reagent mixture has improved filmity, gas transparency, photobleaching characteristics and solubility in alkaline which are essential to the device fabrication. Submicron photoresist patterns are successfully fabricated by a simple sequence of photolithography process. The WSP layer has been applied to the bilayer resist system--deep-UV portable conformable masking (PCM)--that is not affected by VLSI's topography, and is able to fabricate highly accurate pattern. The aqueous developable layer, PMGI, with high organic solvent resistance is used in the bottom layer. Therefore, no interfacial mixing with conventional positive resist top layer is observed. Furthermore, deep-UV exposure method has been used for the KrF excimer laser optical system in order to increase high throughput. From the experiments, it has been confirmed that good resist transfer profile can be realized by the use of WSP, and that the submicron resist patterns with high aspect-ratio can be developed on the nonplaner wafer with steps of up to 41m by the combination of the WSP with the PCM system. By this technology, has been improved the weak point: variation in the line width due to the thickness of contrast-enhanced layer when the CEL technology is applied, and dependency of both the finished resist profile and the line-width accuracy on the thickness of the top layer resist when the PCM system is adopted.

  11. Detecting Intramolecular Conformational Dynamics of Single Molecules in Short Distance Range with Sub-Nanometer Sensitivity

    PubMed Central

    Zhou, Ruobo; Kunzelmann, Simone; Webb, Martin R.; Ha, Taekjip

    2011-01-01

    Single molecule detection is useful for characterizing nanoscale objects such as biological macromolecules, nano-particles and nano-devices with nano-meter spatial resolution. Fluorescence resonance energy transfer (FRET) is widely used as a single-molecule assay to monitor intramolecular dynamics in the distance range of 3–8 nm. Here we demonstrate that self-quenching of two rhodamine derivatives can be used to detect small conformational dynamics corresponding to sub-nanometer distance changes in a FRET-insensitive short range at the single molecule level. A ParM protein mutant labeled with two rhodamines works as a single molecule ADP sensor which has 20 times brighter fluorescence signal in the ADP bound state than the unbound state. Single molecule time trajectories show discrete transitions between fluorescence on and off states that can be directly ascribed to ADP binding and dissociation events. The conformational changes observed with 20:1 contrast are only 0.5 nm in magnitude and are between crystallographic distances of 1.6 nm and 2.1 nm, demonstrating exquisite sensitivity to short distance scale changes. The systems also allowed us to gain information on the photophysics of self-quenching induced by rhodamine stacking: (1) photobleaching of either of the two rhodamines eliminates quenching of the other rhodamine fluorophore and (2) photobleaching from the highly quenched, stacked state is only two-fold slower than from the unstacked state. PMID:22023515

  12. Minimizing ATP depletion by oxygen scavengers for single-molecule fluorescence imaging in live cells.

    PubMed

    Jung, Seung-Ryoung; Deng, Yi; Kushmerick, Christopher; Asbury, Charles L; Hille, Bertil; Koh, Duk-Su

    2018-06-19

    The stability of organic dyes against photobleaching is critical in single-molecule tracking and localization microscopy. Since oxygen accelerates photobleaching of most organic dyes, glucose oxidase is commonly used to slow dye photobleaching by depleting oxygen. As demonstrated here, pyranose-2-oxidase slows bleaching of Alexa647 dye by ∼20-fold. However, oxygen deprivation may pose severe problems for live cells by reducing mitochondrial oxidative phosphorylation and ATP production. We formulate a method to sustain intracellular ATP levels in the presence of oxygen scavengers. Supplementation with metabolic intermediates including glyceraldehyde, glutamine, and α-ketoisocaproate maintained the intracellular ATP level for at least 10 min by balancing between FADH 2 and NADH despite reduced oxygen levels. Furthermore, those metabolites supported ATP-dependent synthesis of phosphatidylinositol 4,5-bisphosphate and internalization of PAR2 receptors. Our method is potentially relevant to other circumstances that involve acute drops of oxygen levels, such as ischemic damage in the brain or heart or tissues for transplantation.

  13. Experts' Understanding of Partial Derivatives Using the Partial Derivative Machine

    ERIC Educational Resources Information Center

    Roundy, David; Weber, Eric; Dray, Tevian; Bajracharya, Rabindra R.; Dorko, Allison; Smith, Emily M.; Manogue, Corinne A.

    2015-01-01

    Partial derivatives are used in a variety of different ways within physics. Thermodynamics, in particular, uses partial derivatives in ways that students often find especially confusing. We are at the beginning of a study of the teaching of partial derivatives, with a goal of better aligning the teaching of multivariable calculus with the needs of…

  14. Photodynamic activity of pyropheophorbide methyl ester and pyropheophorbide a in dimethylformamide solution.

    PubMed

    Al-Omari, Saleh; Ali, Ahmad

    2009-03-01

    Comparative spectroscopic study including the photosensitizers of pyropheophorbide methyl ester (PPME) and pyropheophorbide a (PPa) was performed to study their photodynamic activity. The investigated photosensitizers in a homogeneous system of dimethylformamide (DMF) are not photostable upon irradiation. The photobleaching efficiency of PPa is higher than that of PPME. Combining these results with the data obtained by measuring the singlet oxygen quantum yield and the hydroxyl group generation, it was revealed that the photobleaching efficiency could be correlated with the singlet oxygen quantum yield and the hydroxyl group production of the photosensitizer.

  15. 21 CFR 184.1685 - Rennet (animal-derived) and chymosin preparation (fermentation-derived).

    Code of Federal Regulations, 2010 CFR

    2010-04-01

    ... (fermentation-derived). 184.1685 Section 184.1685 Food and Drugs FOOD AND DRUG ADMINISTRATION, DEPARTMENT OF... (animal-derived) and chymosin preparation (fermentation-derived). (a)(1) Rennet and bovine rennet are... clear solution containing the active enzyme chymosin (E.C. 3.4.23.4). It is derived, via fermentation...

  16. Endohedral Metallofullerene Derivatives

    NASA Technical Reports Server (NTRS)

    Dorn, Harry C. (Inventor); Iezzi, Erick B. (Inventor); Duchamp, James (Inventor)

    2008-01-01

    Trimetallic nitride endohedral metallofullerene derivatives and their preparation are described. The trimetallic nitride endohedral metallofullerene derivatives have the general formula A(sub 3-n)X(sub n)@C(sub m)(R) where n ranges from 0 to 3, A and X may be trivalent metals and may be either rare earth metal or group IIIB metals, m is between about 60 and about 200, and R is preferably an organic group. Derivatives where the R group forms cyclized derivatives with the fullerene cage are also described.

  17. Derivation in INK-algebras

    NASA Astrophysics Data System (ADS)

    Kaviyarasu, M.; Indhira, K.

    2018-04-01

    In 2017 we introduced a new notion of algebra called IKN-algebra. Motivated by some result on derivations (rightleft)-derivation and (leftright)- derivation in ring. In this paper we introduce derivation in INK-Algebras and investigate some important result.

  18. Derivative chameleons

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

    Noller, Johannes, E-mail: johannes.noller08@imperial.ac.uk

    2012-07-01

    We consider generalized chameleon models where the conformal coupling between matter and gravitational geometries is not only a function of the chameleon field φ, but also of its derivatives via higher order co-ordinate invariants (such as ∂{sub μ}φ∂{sup μ}φ,□φ,...). Specifically we consider the first such non-trivial conformal factor A(φ,∂{sub μ}φ∂{sup μ}φ). The associated phenomenology is investigated and we show that such theories have a new generic mass-altering mechanism, potentially assisting the generation of a sufficiently large chameleon mass in dense environments. The most general effective potential is derived for such derivative chameleon setups and explicit examples are given. Interestingly thismore » points us to the existence of a purely derivative chameleon protected by a shift symmetry for φ → φ+c. We also discuss potential ghost-like instabilities associated with mass-lifting mechanisms and find another, mass-lowering and instability-free, branch of solutions. This suggests that, barring fine-tuning, stable derivative models are in fact typically anti-chameleons that suppress the field's mass in dense environments. Furthermore we investigate modifications to the thin-shell regime and prove a no-go theorem for chameleon effects in non-conformal geometries of the disformal type.« less

  19. 40 CFR 721.10039 - Diethoxybenzenamine derivative, diazotized, coupled with aminonaphthalenesulfonic acid derivative...

    Code of Federal Regulations, 2011 CFR

    2011-07-01

    ... 40 Protection of Environment 31 2011-07-01 2011-07-01 false Diethoxybenzenamine derivative, diazotized, coupled with aminonaphthalenesulfonic acid derivative, ammonium salt (generic). 721.10039 Section... Substances § 721.10039 Diethoxybenzenamine derivative, diazotized, coupled with aminonaphthalenesulfonic acid...

  20. 40 CFR 721.10039 - Diethoxybenzenamine derivative, diazotized, coupled with aminonaphthalenesulfonic acid derivative...

    Code of Federal Regulations, 2010 CFR

    2010-07-01

    ... 40 Protection of Environment 30 2010-07-01 2010-07-01 false Diethoxybenzenamine derivative, diazotized, coupled with aminonaphthalenesulfonic acid derivative, ammonium salt (generic). 721.10039 Section... Substances § 721.10039 Diethoxybenzenamine derivative, diazotized, coupled with aminonaphthalenesulfonic acid...

  1. FLIPing heterokaryons to analyze nucleo-cytoplasmic shuttling of yeast proteins.

    PubMed

    Belaya, Katsiaryna; Tollervey, David; Kos, Martin

    2006-05-01

    Nucleo-cytoplasmic shuttling is an important feature of proteins involved in nuclear export/import of RNAs, proteins, and also large ribonucleoprotein complexes such as ribosomes. The vast amount of proteomic data available shows that many of these processes are highly dynamic. Therefore, methods are needed to reliably assess whether a protein shuttles between nucleus and cytoplasm, and the kinetics with which it exchanges. Here we describe a combination of the classical heterokaryon assay with fluorescence recovery after photobleaching (FRAP) and fluorescence loss in photobleaching (FLIP) techniques, which allows an assessment of the kinetics of protein shuttling in the yeast Saccharomyces cerevisiae.

  2. How important are intertidal ecosystems for global biogeochemical cycles? Molecular and isotopic evidence for major outwelling of photo-bleached dissolved organic matter from mangroves.

    NASA Astrophysics Data System (ADS)

    Dittmar, T.; Cooper, W. T.; Koch, B. P.; Kattner, G.

    2006-05-01

    Organic matter, which is dissolved in low concentrations in the vast waters of the oceans, contains a total amount of carbon similar to atmospheric carbon dioxide. To understand global biogeochemical cycles it is crucial to quantify the sources of marine dissolved organic carbon (DOC). We investigated the impact of mangroves, the dominant intertidal vegetation of the tropics, on marine DOC inventories. Stable carbon- isotopes, ultrahigh-resolution mass spectrometry (FTICRMS), lignin-derived phenols and proton nuclear magnetic resonance spectroscopy showed that mangroves are the main source of terrigenous DOC on the shelf off Northern Brazil. Sunlight efficiently destroyed aromatic molecules during transport offshore, removing about one third of mangrove-derived DOC. The remainder was refractory and may thus be distributed over the oceans. On a global scale, we estimate that mangroves account for more than 10 percent of the terrestrially- derived, refractory DOC transported to the ocean, while they cover less than 0.1 percent of the continents' surface.

  3. Uncovering the Design Principle of Amino Acid-Derived Photoluminescent Biodots with Tailor-Made Structure-Properties and Applications for Cellular Bioimaging.

    PubMed

    Xu, Hesheng Victor; Zheng, Xin Ting; Zhao, Yanli; Tan, Yen Nee

    2018-06-13

    Natural amino acids possess side chains with different functional groups (R groups), which make them excellent precursors for programmable synthesis of biomolecule-derived nanodots (biodots) with desired properties. Herein, we report the first systematic study to uncover the material design rules of biodot synthesis from 20 natural α-amino acids via a green hydrothermal approach. The as-synthesized amino acid biodots (AA dots) are comprehensively characterized to establish a structure-property relationship between the amino acid precursors and the corresponding photoluminescent properties of AA dots. It was found that the amino acids with reactive R groups, including amine, hydroxyl, and carboxyl functional groups form unique C-O-C/C-OH and N-H bonds in the AA dots which stabilize the surface defects, giving rise to brightly luminescent AA dots. Furthermore, the AA dots were found to be amorphous and the length of the R group was observed to affect the final morphology (e.g., disclike nanostructure, nanowire, or nanomesh) of the AA dots, which in turn influence their photoluminescent properties. It is noteworthy to highlight that the hydroxyl-containing amino acids, that is, Ser and Thr, form the brightest AA dots with a quantum yield of 30.44% and 23.07%, respectively, and possess high photostability with negligible photobleaching upon continuous UV exposure for 3 h. Intriguingly, by selective mixing of Ser or Thr with another amino acid precursor, the resulting mixed AA dots could inherit unique properties such as improved photostability and significant red shift in their emission wavelength, producing enhanced green and red fluorescent intensity. Moreover, our cellular studies demonstrate that the as-synthesized AA dots display outstanding biocompatibility and excellent intracellular uptake, which are highly desirable for imaging applications. We envision that the material design rules discovered in this study will be broadly applicable for the rational

  4. Photounbinding of Calmodulin from a Family of CaM Binding Peptides

    PubMed Central

    Neumüller, Klaus G.; Elsayad, Kareem; Reisecker, Johannes M.; Waxham, M. Neal; Heinze, Katrin G.

    2010-01-01

    Background Recent studies have shown that fluorescently labeled antibodies can be dissociated from their antigen by illumination with laser light. The mechanism responsible for the photounbinding effect, however, remains elusive. Here, we give important insights into the mechanism of photounbinding and show that the effect is not restricted to antibody/antigen binding. Methodology/Principal Findings We present studies of the photounbinding of labeled calmodulin (CaM) from a set of CaM-binding peptides with different affinities to CaM after one- and two-photon excitation. We found that the photounbinding effect becomes stronger with increasing binding affinity. Our observation that photounbinding can be influenced by using free radical scavengers, that it does not occur with either unlabeled protein or non-fluorescent quencher dyes, and that it becomes evident shortly after or with photobleaching suggest that photounbinding and photobleaching are closely linked. Conclusions/Significance The experimental results exclude surface effects, or heating by laser irradiation as potential causes of photounbinding. Our data suggest that free radicals formed through photobleaching may cause a conformational change of the CaM which lowers their binding affinity with the peptide or its respective binding partner. PMID:21124984

  5. High-sensitivity detection of biological amines using fast Hadamard transform CE coupled with photolytic optical gating.

    PubMed

    Braun, Kevin L; Hapuarachchi, Suminda; Fernandez, Facundo M; Aspinwall, Craig A

    2007-08-01

    Here, we report the first utilization of Hadamard transform CE (HTCE), a high-sensitivity, multiplexed CE technique, with photolytic optical gating sample injection of caged fluorescent labels for the detection of biologically important amines. Previous implementations of HTCE have relied upon photobleaching optical gating sample injection of fluorescent dyes. Photolysis of caged fluorescent labels reduces the fluorescence background, providing marked enhancements in sensitivity compared to photobleaching. Application of fast Hadamard transform CE (fHTCE) for fluorescein-based dyes yields a ten-fold higher sensitivity for photolytic injections compared to photobleaching injections, due primarily to the reduced fluorescent background provided by caged fluorescent dyes. Detection limits as low as 5 pM (ca. 18 molecules per injection event) were obtained with on-column LIF detection using fHTCE in less than 25 s, with the capacity for continuous, online separations. Detection limits for glutamate and aspartate below 150 pM (1-2 amol/injection event) were obtained using photolytic sample injection, with separation efficiencies exceeding 1 x 10(6) plates/m and total multiplexed separation times as low as 8 s. These results strongly support the feasibility of this approach for high-sensitivity dynamic chemical monitoring applications.

  6. Implicit dosimetry of microorganism photodynamic inactivation

    NASA Astrophysics Data System (ADS)

    Tamošiūnas, Mindaugas; Kuliešienė, Neringa; Daugelavičius, Rimantas

    2017-12-01

    Photosensitization based antibacterial treatment is efficient against a broad range of pathogens but it utilizes suboptimal dosimetry with an explicit (and very broad range) determination of sensitizer concentration, light dose and fluence rates. In this study we verified the implicit dosimetry approach for pathogen photodynamic treatment, employing protoporphyrin IX (ppIX) photobleaching to assess the killing efficacy against Staphylococcus aureus and Candida albicans cells. The results show that there was an increased kill of S. aureus and C. albicans at higher degree of ppIX fluorescence decay. Therefore ppIX photobleaching can be incorporated into the PDI dose metric offering to predict the pathogen killing efficacy during photodynamic treatment.

  7. Strong plasmonic enhancement of single molecule photostability in silver dimer optical antennas

    NASA Astrophysics Data System (ADS)

    Kaminska, Izabela; Vietz, Carolin; Cuartero-González, Álvaro; Tinnefeld, Philip; Fernández-Domínguez, Antonio I.; Acuna, Guillermo P.

    2018-02-01

    Photobleaching is an effect terminating the photon output of fluorophores, limiting the duration of fluorescence-based experiments. Plasmonic nanoparticles (NPs) can increase the overall fluorophore photostability through an enhancement of the radiative rate. In this work, we use the DNA origami technique to arrange a single fluorophore in the 12-nm gap of a silver NP dimer and study the number of emitted photons at the single molecule level. Our findings yielded a 30× enhancement in the average number of photons emitted before photobleaching. Numerical simulations are employed to rationalize our results. They reveal the effect of silver oxidation on decreasing the radiative rate enhancement.

  8. Real-Time Dosimetry and Optimization of Prostate Photodynamic Therapy

    DTIC Science & Technology

    2005-05-01

    85cm’ 0:k -" l)=O5.0O25,=0.10cnf’ Ps4 ൱ crm’ 0 cik)=5.012p 5,4=5.45c,1’. G~cff’* - 0~n) .012,P. 0.49cff’,0:=3.56c-0 cink) = - 1,00 c ’", V-= 1.75 ccml...tissue to make a measurement. This limits the resolution of absorption spectroscopy, and makes it more time- consuming than fluorescence spectroscopy...photobleaching versus those that do not. Further research into the photobleaching behavior and in vivo vascular effects of MLu is needed to resolve the

  9. Immobilization of zinc phthalocyanines in silicate matrices and investigation of their photobactericidal effect on E. coli.

    PubMed

    Artarsky, Spas; Dimitrova, Stanislava; Bonnett, Raymond; Krysteva, Milka

    2006-03-26

    The aim of the present investigation was to immobilize zinc phthalocyanines in a silicate matrix and to test the photobactericidal properties of the matrices so prepared toward Esherichia coli in model aqueous media. For the purpose, tetra tertiary butyl zinc phthalocyanine (TBZnPc) and zinc phthalocyanine tetrasulfonic acid (ZnPcTS) were used. The abilities of these two photosensitizers to generate singlet oxygen in solution were compared by following the rate of photobleaching of 1,3-diphenylisobenzofuran (DPBF) at 430 nm in dimethylformamide (DMF). The results of this study show clearly that, under the conditions used here, the TBZnPc is the more effective generator of singlet oxygen; with it the DPBF was virtually completely photobleached in 4 min, while with the ZnPcTS under the same conditions, it took 12 min to reach this point. Glass conjugates with the two phthalocyanines were obtained by the sol-gel technique and were characterized by a well-defined color due to the phthalocyanine incorporated in the silicate matrix. Glasses with an intense, but inhomogeneous, green color were obtained when the tetrasulfonic derivative of the zinc phthalocyanine was used, while blue glasses of evenly distributed coloration were formed from the tetra tertiary butyl derivative. The ZnPcTS conjugate demonstrates more effective singlet oxygen evolution than is the case with the TBZnPc conjugate. These results are the opposite of those obtained for the free phthalocyanines in solution. The structural formulae of the compounds show that TBZnPc has a more pronounced hydrophobic character than the sulfonic derivative. In our view, the relative reactivities of the conjugates can be explained by the tetrasulfonic derivative being situated mainly in the surface parts of the glass matrix where the hydrophilic character is prevailing, while the tertiary butyl derivative is mainly present in the internal parts of the matrix as a result of which it is less accessible and therefore less

  10. 78 FR 32191 - Derivatives

    Federal Register 2010, 2011, 2012, 2013, 2014

    2013-05-29

    ...This proposed rule permits credit unions to engage in limited derivatives activities for the purpose of mitigating interest rate risk. This proposed rule applies to federal credit unions and any federally insured, state-chartered credit unions that are permitted under applicable state law to engage in derivatives transactions. It requires any credit union seeking derivatives authority to submit an application for one of two levels of authority. Level I and Level II authority differ on the permissible levels of transactions as well as the application, expertise, and systems requirements associated with operating a derivatives program.

  11. Antioxidant and intestinal anti-inflammatory effects of plant-derived coumarin derivatives.

    PubMed

    Witaicenis, Aline; Seito, Leonardo Noboru; da Silveira Chagas, Alexandre; de Almeida, Luiz Domingues; Luchini, Ana Carolina; Rodrigues-Orsi, Patrícia; Cestari, Silvia Helena; Di Stasi, Luiz Claudio

    2014-02-15

    Coumarins, also known as benzopyrones, are plant-derived products with several pharmacological properties, including antioxidant and anti-inflammatory activities. Based on the wide distribution of coumarin derivatives in plant-based foods and beverages in the human diet, our objective was to evaluate both the antioxidant and intestinal anti-inflammatory activities of six coumarin derivatives of plant origin (scopoletin, scoparone, fraxetin, 4-methyl-umbeliferone, esculin and daphnetin) to verify if potential intestinal anti-inflammatory activity was related to antioxidant properties. Intestinal inflammation was induced by intracolonic instillation of TNBS in rats. The animals were treated with coumarins by oral route. The animals were killed 48 h after colitis induction. The colonic segments were obtained after laparotomy and macroscopic and biochemical parameters (determination of glutathione level and myeloperoxidase and alkaline phosphatase activities) were evaluated. The antioxidant properties of these coumarins were examined by lipid peroxidation and DPPH assays. Treatment with esculin, scoparone and daphnetin produced the best protective effects. All coumarin derivatives showed antioxidant activity in the DPPH assay, while daphnetin and fraxetin also showed antioxidant activity by inhibiting lipid peroxidation. Coumarins, except 4-methyl-umbeliferone, also showed antioxidant activity through the counteraction of glutathione levels or through the inhibition of myeloperoxidase activity. The intestinal anti-inflammatory activity of coumarin derivatives were related to their antioxidant properties, suggesting that consumption of coumarins and/or foods rich in coumarin derivatives, particularly daphnetin, esculin and scoparone, could prevent intestinal inflammatory disease. Copyright © 2013 Elsevier GmbH. All rights reserved.

  12. Coastal conduit in southwestern Hudson Bay (Canada) in summer: Rapid transit of freshwater and significant loss of colored dissolved organic matter

    NASA Astrophysics Data System (ADS)

    Granskog, Mats A.; MacDonald, Robie W.; Kuzyk, Zou Zou A.; Senneville, Simon; Mundy, Christopher-John; Barber, David G.; Stern, Gary A.; Saucier, Francois

    2009-08-01

    Distributions of freshwater (sea-ice melt and runoff) were investigated along inshore-offshore sections in southwestern Hudson Bay for fall conditions. Conductivity-temperature-density profiles and bottle samples collected for salinity, oxygen isotope (δ18O), and colored dissolved organic matter (CDOM) analyses were used to discriminate between contributions of river water (RW) and sea-ice melt (SIM). Stations had a fresh summer surface mixed layer 5-25 m thick overlying a cold subsurface layer indicative of the previous winter's polar mixed layer (PML). The fraction of RW decreased strongly with distance from shore, while the opposite was true for SIM. The majority of RW was constrained in a coastal domain within 100-150 km from shore, which, because of high alongshore velocities, accounts for the majority of freshwater and volume transports. On the basis of freshwater inventories and composition, brine and RW accumulate in the PML over winter because of ice formation and downward mixing. The summer surface circulation results in an annual net export of SIM from the region. Residence times for freshwater components in the southwestern sector of the bay, based on currents derived from a 3-D ocean model for Hudson Bay, are about 1-10 months, implying rapid transit of freshwater. Despite the short residence time for RW (1-3 months), CDOM is significantly photobleached and provides an unreliable tracer for RW. Photobleaching represents an important sink for dissolved organic carbon entering from rivers and could, in part, explain why Hudson Bay is only a minor sink for atmospheric CO2 in the open water season.

  13. Potential of BODIPY-cholesterol for analysis of cholesterol transport and diffusion in living cells.

    PubMed

    Wüstner, Daniel; Lund, Frederik W; Röhrl, Clemens; Stangl, Herbert

    2016-01-01

    Cholesterol is an abundant and important lipid component of cellular membranes. Analysis of cholesterol transport and diffusion in living cells is hampered by the technical challenge of designing suitable cholesterol probes which can be detected for example by optical microscopy. One strategy is to use intrinsically fluorescent sterols, as dehydroergosterol (DHE), having minimal chemical alteration compared to cholesterol but giving low fluorescence signals in the UV region of the spectrum. Alternatively, one can use dye-tagged cholesterol analogs and in particular BODIPY-cholesterol (BChol), whose synthesis and initial characterization was pioneered by Robert Bittman. Here, we give a general overview of the properties and applications but also limitations of BODIPY-tagged cholesterol probes for analyzing intracellular cholesterol trafficking. We describe our own experiences and collaborative efforts with Bob Bittman for studying diffusion in the plasma membrane (PM) and uptake of BChol in a quantitative manner. For that purpose, we used a variety of fluorescence approaches including fluorescence correlation spectroscopy and its imaging variants, fluorescence recovery after photobleaching (FRAP) and fluorescence loss in photobleaching (FLIP). We also describe pulse-chase studies from the PM using BChol in direct comparison to DHE. Based on the gathered imaging data, we present a two-step kinetic model for sterol transport between PM and recycling endosomes. In addition, we highlight the suitability of BChol for determining transport of lipoprotein-derived sterol using electron microscopy (EM) and show that this approach ideally complements fluorescence studies. Copyright © 2015 Elsevier Ireland Ltd. All rights reserved.

  14. Cytoplasmic Electric Fields and Electroosmosis: Possible Solution for the Paradoxes of the Intracellular Transport of Biomolecules

    PubMed Central

    Andreev, Victor P.

    2013-01-01

    The objective of the paper is to show that electroosmotic flow might play an important role in the intracellular transport of biomolecules. The paper presents two mathematical models describing the role of electroosmosis in the transport of the negatively charged messenger proteins to the negatively charged nucleus and in the recovery of the fluorescence after photobleaching. The parameters of the models were derived from the extensive review of the literature data. Computer simulations were performed within the COMSOL 4.2a software environment. The first model demonstrated that the presence of electroosmosis might intensify the flux of messenger proteins to the nucleus and allow the efficient transport of the negatively charged phosphorylated messenger proteins against the electrostatic repulsion of the negatively charged nucleus. The second model revealed that the presence of the electroosmotic flow made the time of fluorescence recovery dependent on the position of the bleaching spot relative to cellular membrane. The magnitude of the electroosmotic flow effect was shown to be quite substantial, i.e. increasing the flux of the messengers onto the nucleus up to 4-fold relative to pure diffusion and resulting in the up to 3-fold change in the values of fluorescence recovery time, and therefore the apparent diffusion coefficient determined from the fluorescence recovery after photobleaching experiments. Based on the results of the modeling and on the universal nature of the electroosmotic flow, the potential wider implications of electroosmotic flow in the intracellular and extracellular biological processes are discussed. Both models are available for download at ModelDB. PMID:23613967

  15. Cytoplasmic electric fields and electroosmosis: possible solution for the paradoxes of the intracellular transport of biomolecules.

    PubMed

    Andreev, Victor P

    2013-01-01

    The objective of the paper is to show that electroosmotic flow might play an important role in the intracellular transport of biomolecules. The paper presents two mathematical models describing the role of electroosmosis in the transport of the negatively charged messenger proteins to the negatively charged nucleus and in the recovery of the fluorescence after photobleaching. The parameters of the models were derived from the extensive review of the literature data. Computer simulations were performed within the COMSOL 4.2a software environment. The first model demonstrated that the presence of electroosmosis might intensify the flux of messenger proteins to the nucleus and allow the efficient transport of the negatively charged phosphorylated messenger proteins against the electrostatic repulsion of the negatively charged nucleus. The second model revealed that the presence of the electroosmotic flow made the time of fluorescence recovery dependent on the position of the bleaching spot relative to cellular membrane. The magnitude of the electroosmotic flow effect was shown to be quite substantial, i.e. increasing the flux of the messengers onto the nucleus up to 4-fold relative to pure diffusion and resulting in the up to 3-fold change in the values of fluorescence recovery time, and therefore the apparent diffusion coefficient determined from the fluorescence recovery after photobleaching experiments. Based on the results of the modeling and on the universal nature of the electroosmotic flow, the potential wider implications of electroosmotic flow in the intracellular and extracellular biological processes are discussed. Both models are available for download at ModelDB.

  16. Studies of porphyrin-containing specimens using an optical spectrometer connected to a confocal scanning laser microscope.

    PubMed

    Trepte, O; Rokahr, I; Andersson-Engels, S; Carlsson, K

    1994-12-01

    A spectrometer has been developed for use with a confocal scanning laser microscope. With this unit, spectral information from a single point or a user-defined region within the microscope specimen can be recorded. A glass prism is used to disperse the spectral components of the recorded light over a linear CCD photodiode array with 256 elements. A regulated cooling unit keeps the detector at 277 K, thereby allowing integration times of up to 60 s. The spectral resolving power, lambda/delta lambda, ranges from 350 at lambda = 400 nm to 100 at lambda = 700 nm. Since the entrance aperture of the spectrometer has the same size as the detector pinhole used during normal confocal scanning, the three-dimensional spatial resolution is equivalent to that of normal confocal scanning. Light from the specimen is deflected to the spectrometer by a solenoid controlled mirror, allowing fast and easy switching between normal confocal scanning and spectrometer readings. With this equipment, studies of rodent liver specimens containing porphyrins have been made. The subcellular localization is of interest for the mechanisms of photodynamic therapy (PDT) of malignant tumours. Spectroscopic detection is necessary to distinguish the porphyrin signal from other fluorescent components in the specimen. Two different substances were administered to the tissue, Photofrin, a haematoporphyrin derivative (HPD) and delta-amino levulinic acid (ALA), a precursor to protoporphyrin IX and haem in the haem cycle. Both are substances under clinical trials for PDT of malignant tumours. Following administration of these compounds to the tissue, the potent photosensitizer and fluorescent compound Photofrin, or protoporphyrin IX, respectively, is accumulated.(ABSTRACT TRUNCATED AT 250 WORDS)

  17. Certain legal aspects of derivatives.

    PubMed

    Cloney, T J

    1994-01-01

    This article discusses legal characteristics of derivatives that a not-for-profit health care company may use in a conduit financing in which a governmental issuer issues bonds for the benefit of a health care company. This article also presents the Master Agreement promulgated by the International Swaps and Derivatives Association; discusses how to get out of a derivative; discusses the status of a derivative after insolvency of a counterparty; presents disclosure issues relating to documents prepared in connection with an issue of bonds; and describes the treatment of derivatives under commodities regulations. It concludes with a presentation of possible new regulations applying to derivatives.

  18. Analytic computation of energy derivatives - Relationships among partial derivatives of a variationally determined function

    NASA Technical Reports Server (NTRS)

    King, H. F.; Komornicki, A.

    1986-01-01

    Formulas are presented relating Taylor series expansion coefficients of three functions of several variables, the energy of the trial wave function (W), the energy computed using the optimized variational wave function (E), and the response function (lambda), under certain conditions. Partial derivatives of lambda are obtained through solution of a recursive system of linear equations, and solution through order n yields derivatives of E through order 2n + 1, extending Puley's application of Wigner's 2n + 1 rule to partial derivatives in couple perturbation theory. An examination of numerical accuracy shows that the usual two-term second derivative formula is less stable than an alternative four-term formula, and that previous claims that energy derivatives are stationary properties of the wave function are fallacious. The results have application to quantum theoretical methods for the computation of derivative properties such as infrared frequencies and intensities.

  19. Influence of severe drought conditions on chromophoric dissolved organic matter dynamics in south Texas coastal waters

    NASA Astrophysics Data System (ADS)

    Miller, Stephanie A.; Shank, G. Christopher

    2013-01-01

    We investigated chromophoric dissolved organic matter (CDOM) dynamics in response to the severe drought conditions of 2009 in the Aransas-Copano (AC) Bay complex and adjacent nearshore Gulf of Mexico (GoM) along the southern Texas (USA) coast. Surface water absorption coefficients (a305 in m-1) were measured daily at the University of Texas at Austin Marine Science Institute (UTMSI) pier located at the interface between the AC complex and the GoM. From June to August 2009, a305 averaged 0.81 ± 0.31 m-1 compared to 1.20 ± 0.70 m-1 during summer of 2008 (June-August) and 1.84 ± 0.34 m-1 during summer 2007 (non-drought year). Despite negligible freshwater input to the AC system for most of 2009, mean a305 values for AC Bay sites were similar in 2008 and 2009, ranging from 3.24 ± 0.60 m-1 in the lower estuary to 6.22 ± 0.55 m-1 in the upper estuary during summer 2009, and 3.15 ± 0.38 m-1 to 6.81 ± 0.73 m-1 during summer 2008 (no data for 2007). CDOM photobleaching experiments were performed using a SunTest XLS + solar simulator with an irradiation spectrum that closely matches solar UV at subtropical latitudes. AC Bay samples exhibited a305 photobleaching half-lives of 33-48 h, while GoM samples exhibited a305 photobleaching half-lives of 36-89 h. We estimate that summertime photobleaching may reduce CDOM levels by >50% in the AC bay complex before discharge into the GoM and by up to 25% more throughout the nearshore waters along the western GoM shelf.

  20. Light and Life in Baltimore—and Beyond

    PubMed Central

    Edidin, Michael

    2015-01-01

    Baltimore has been the home of numerous biophysical studies using light to probe cells. One such study, quantitative measurement of lateral diffusion of rhodopsin, set the standard for experiments in which recovery after photobleaching is used to measure lateral diffusion. Development of this method from specialized microscopes to commercial scanning confocal microscopes has led to widespread use of the technique to measure lateral diffusion of membrane proteins and lipids, and as well diffusion and binding interactions in cell organelles and cytoplasm. Perturbation of equilibrium distributions by photobleaching has also been developed into a robust method to image molecular proximity in terms of fluorescence resonance energy transfer between donor and acceptor fluorophores. PMID:25650914

  1. PhotoGate microscopy: tracking single molecules in a cytoplasm (Conference Presentation)

    NASA Astrophysics Data System (ADS)

    Yildiz, Ahmet

    2016-02-01

    Tracking single molecules inside cells reveals the dynamics of biological processes, including receptor trafficking, signaling and cargo transport. However, individual molecules often cannot be resolved inside cells due to their high density in the cellular environment. We developed a photobleaching gate assay, which controls the number of fluorescent particles in a region of interest by repeatedly photobleaching its boundary. Using this method, we tracked single particles at surface densities two orders of magnitude higher than the single-molecule detection limit. We observed ligand-induced dimerization of epidermal growth factor receptors (EGFR) on a live cell membrane. In addition, we tracked individual intraflagellar transport (IFT) trains along the length of a cilium and observed their remodeling at the ciliary tip.

  2. Rapid Raman spectroscopy of musculoskeletal tissue using a visible laser and an electron-multiplying CCD (EMCCD) detector

    NASA Astrophysics Data System (ADS)

    Golcuk, Kurtulus; Mandair, Gurjit S.; Callender, Andrew F.; Finney, William F.; Sahar, Nadder; Kohn, David H.; Morris, Michael D.

    2006-02-01

    Background fluorescence can often complicate the use of Raman microspectroscopy in the study of musculoskeletal tissues. Such fluorescence interferences are undesirable as the Raman spectra of matrix and mineral phases can be used to differentiate between normal and pathological or microdamaged bone. Photobleaching with the excitation laser provides a non-invasive method for reducing background fluorescence, enabling 532 nm Raman hyperspectral imaging of bone tissue. The signal acquisition time for a 400 point Raman line image is reduced to 1-4 seconds using electronmultiplying CCD (EMCCD) detector, enabling acquisition of Raman images in less than 10 minutes. Rapid photobleaching depends upon multiple scattering effects in the tissue specimen and is applicable to some, but not all experimental situations.

  3. Effect of chondrocyte-derived early extracellular matrix on chondrogenesis of placenta-derived mesenchymal stem cells.

    PubMed

    Park, Yong-Beom; Seo, Sinji; Kim, Jin-A; Heo, Jin-Chul; Lim, Young-Cheol; Ha, Chul-Won

    2015-06-24

    The extracellular matrix (ECM) surrounding cells contains a variety of proteins that provide structural support and regulate cellular functions. Previous studies have shown that decellularized ECM isolated from tissues or cultured cells can be used to improve cell differentiation in tissue engineering applications. In this study we evaluated the effect of decellularized chondrocyte-derived ECM (CDECM) on the chondrogenesis of human placenta-derived mesenchymal stem cells (hPDMSCs) in a pellet culture system. After incubation with or without chondrocyte-derived ECM in chondrogenic medium for 1 or 3 weeks, the sizes and wet masses of the cell pellets were compared with untreated controls (hPDMSCs incubated in chondrogenic medium without chondrocyte-derived ECM). In addition, histologic analysis of the cell pellets (Safranin O and collagen type II staining) and quantitative reverse transcription-PCR analysis of chondrogenic markers (aggrecan, collagen type II, and SOX9) were carried out. Our results showed that the sizes and masses of hPDMSC pellets incubated with chondrocyte-derived ECM were significantly higher than those of untreated controls. Differentiation of hPDMSCs (both with and without chondrocyte-derived ECM) was confirmed by Safranin O and collagen type II staining. Chondrogenic marker expression and glycosaminoglycan (GAG) levels were significantly higher in hPDMSC pellets incubated with chondrocyte-derived ECM compared with untreated controls, especially in cells precultured with chondrocyte-derived ECM for 7 d. Taken together, these results demonstrate that chondrocyte-derived ECM enhances the chondrogenesis of hPDMSCs, and this effect is further increased by preculture with chondrocyte-derived ECM. This preculture method for hPDMSC chondrogenesis represents a promising approach for cartilage tissue engineering.

  4. Monte Carlo modeling of in vivo protoporphyrin IX fluorescence and singlet oxygen production during photodynamic therapy for patients presenting with superficial basal cell carcinomas

    NASA Astrophysics Data System (ADS)

    Valentine, Ronan M.; Brown, C. Tom A.; Moseley, Harry; Ibbotson, Sally; Wood, Kenny

    2011-04-01

    We present protoporphyrin IX (PpIX) fluorescence measurements acquired from patients presenting with superficial basal cell carcinoma during photodynamic therapy (PDT) treatment, facilitating in vivo photobleaching to be monitored. Monte Carlo (MC) simulations, taking into account photobleaching, are performed on a three-dimensional cube grid, which represents the treatment geometry. Consequently, it is possible to determine the spatial and temporal changes to the origin of collected fluorescence and generated singlet oxygen. From our clinical results, an in vivo photobleaching dose constant, β of 5-aminolaevulinic acid-induced PpIX fluorescence is found to be 14 +/- 1 J/cm2. Results from our MC simulations suggest that an increase from our typical administered treatment light dose of 75-150 J/cm2 could increase the effective PDT treatment initially achieved at a depth of 2.7-3.3 mm in the tumor, respectively. Moreover, this increase reduces the surface PpIX fluorescence from 0.00012 to 0.000003 of the maximum value recorded before treatment. The recommendation of administrating a larger light dose, which advocates an increase in the treatment time after surface PpIX fluorescence has diminished, remains valid for different sets of optical properties and therefore should have a beneficial outcome on the total treatment effect.

  5. Alternatives to Outdoor Daylight Illumination for Photodynamic Therapy--Use of Greenhouses and Artificial Light Sources.

    PubMed

    Lerche, Catharina M; Heerfordt, Ida M; Heydenreich, Jakob; Wulf, Hans Christian

    2016-02-29

    Daylight-mediated photodynamic therapy (daylight PDT) is a simple and pain free treatment of actinic keratoses. Weather conditions may not always allow daylight PDT outdoors. We compared the spectrum of five different lamp candidates for indoor "daylight PDT" and investigated their ability to photobleach protoporphyrin IX (PpIX). Furthermore, we measured the amount of PpIX activating daylight available in a glass greenhouse, which can be an alternative when it is uncomfortable for patients to be outdoors. The lamps investigated were: halogen lamps (overhead and slide projector), white light-emitting diode (LED) lamp, red LED panel and lamps used for conventional PDT. Four of the five light sources were able to photobleach PpIX completely. For halogen light and the red LED lamp, 5000 lux could photobleach PpIX whereas 12,000 lux were needed for the white LED lamp. Furthermore, the greenhouse was suitable for daylight PDT since the effect of solar light is lowered only by 25%. In conclusion, we found four of the five light sources and the greenhouse usable for indoor daylight PDT. The greenhouse is beneficial when the weather outside is rainy or windy. Only insignificant ultraviolet B radiation (UVB) radiation passes through the greenhouse glass, so sun protection is not needed.

  6. Alternatives to Outdoor Daylight Illumination for Photodynamic Therapy—Use of Greenhouses and Artificial Light Sources

    PubMed Central

    Lerche, Catharina M.; Heerfordt, Ida M.; Heydenreich, Jakob; Wulf, Hans Christian

    2016-01-01

    Daylight-mediated photodynamic therapy (daylight PDT) is a simple and pain free treatment of actinic keratoses. Weather conditions may not always allow daylight PDT outdoors. We compared the spectrum of five different lamp candidates for indoor “daylight PDT” and investigated their ability to photobleach protoporphyrin IX (PpIX). Furthermore, we measured the amount of PpIX activating daylight available in a glass greenhouse, which can be an alternative when it is uncomfortable for patients to be outdoors. The lamps investigated were: halogen lamps (overhead and slide projector), white light-emitting diode (LED) lamp, red LED panel and lamps used for conventional PDT. Four of the five light sources were able to photobleach PpIX completely. For halogen light and the red LED lamp, 5000 lux could photobleach PpIX whereas 12,000 lux were needed for the white LED lamp. Furthermore, the greenhouse was suitable for daylight PDT since the effect of solar light is lowered only by 25%. In conclusion, we found four of the five light sources and the greenhouse usable for indoor daylight PDT. The greenhouse is beneficial when the weather outside is rainy or windy. Only insignificant ultraviolet B radiation (UVB) radiation passes through the greenhouse glass, so sun protection is not needed. PMID:26938525

  7. Adjoint-based constant-mass partial derivatives

    DOE PAGES

    Favorite, Jeffrey A.

    2017-09-01

    In transport theory, adjoint-based partial derivatives with respect to mass density are constant-volume derivatives. Likewise, adjoint-based partial derivatives with respect to surface locations (i.e., internal interface locations and the outer system boundary) are constant-density derivatives. This study derives the constant-mass partial derivative of a response with respect to an internal interface location or the outer system boundary and the constant-mass partial derivative of a response with respect to the mass density of a region. Numerical results are given for a multiregion two-dimensional (r-z) cylinder for three very different responses: the uncollided gamma-ray flux at an external detector point, k effmore » of the system, and the total neutron leakage. Finally, results from the derived formulas compare extremely well with direct perturbation calculations.« less

  8. Differential interaction of porphyrins used in photoradiation therapy with ferrochelatase.

    PubMed Central

    Dailey, H A; Smith, A

    1984-01-01

    The mechanism of porphyrin accumulation by tumours is not yet established. If metabolism aids porphyrin elimination, tumours, unlike normal tissues, may not metabolize porphyrins used clinically, such as proto-, haemato-, OO'-diacetyl-haemato- and monohydroxyethyl-monovinyl-deutero-porphyrin. Proto-, haemato- and monohydroxyethyl-monovinyl-deutero-porphyrin are substrates for the mitochondrial enzyme ferrochelatase (EC 4.99.1.1), which can form haem analogues from exogenous porphyrins. The Km values for proto-, haemato- and monohydroxyethyl-monovinyl-deutero-porphyrin are 11, 22 and 23 microM respectively. However, OO'-diacetyl-haematoporphyrin is an effective competitive inhibitor with Ki of 11 microM. Hepatic ferrochelatase specific activity is 5.9 and 5.5 nmol of haem/h per mg of protein respectively in normal Buffalo rat and in those bearing the extrahepatic Morris 7288C hepatoma, and is only 0.13 nmol/h per mg in the hepatomas. Therefore low ferrochelatase activity in cancerous cells may provide one means whereby some porphyrins accumulate in tumours, and the ability of certain porphyrins to act as ferrochelatase inhibitors may provide another. PMID:6497856

  9. 12 CFR 403.4 - Derivative classification.

    Code of Federal Regulations, 2010 CFR

    2010-01-01

    ... SAFEGUARDING OF NATIONAL SECURITY INFORMATION § 403.4 Derivative classification. (a) Use of derivative classification. (1) Unlike original classification which is an initial determination, derivative classification... 12 Banks and Banking 4 2010-01-01 2010-01-01 false Derivative classification. 403.4 Section 403.4...

  10. Neuroprotective Ganglioside Derivatives

    DTIC Science & Technology

    2006-09-01

    SH - SY5Y human neuroblastoma cell line . Derivatives determined to have therapeutic potential are tested in vitro for their...to be cytoprotective in in vitro models using the dopaminergic neurotoxin, 1-methyl-4-phenylpyridinium (MPP+) and the SH - SY5Y cell line . Derivatives...action of these two compounds remains unknown. The ability of GM1 (no preincubation) to protect RA- differentiated SH - SY5Y cells from MPP+

  11. Amphetamine derivative related deaths.

    PubMed

    Lora-Tamayo, C; Tena, T; Rodríguez, A

    1997-02-28

    Amphetamine its methylendioxy (methylendioxyamphetamine methylenedioxymethylamphetamine, methylenedioxyethylamphetamine) and methoxy derivatives (p-methoxyamphetamine and p-methoxymethylamphetamine) are widely abused in Spanish society. We present here the results of a systematic study of all cases of deaths brought to the attention of the Madrid department of the Instituto Nacional de Toxicologia from 1993 to 1995 in which some of these drugs have been found in the cadaveric blood. The cases were divided into three categories: amphetamine and derivatives, amphetamines and alcohol, amphetamines and other drugs. Data on age, sex, clinical symptoms, morphological findings, circumstances of death, when known, and concentration of amphetamine derivatives, alcohol and other drugs in blood are given for each group. The information provided here may prove to be useful for the forensic interpretation of deaths which are directly or indirectly related to abuse of amphetamine derivatives.

  12. 3-Nitroasterric Acid Derivatives from an Antarctic Sponge-Derived Pseudogymnoascus sp. Fungus.

    PubMed

    Figueroa, Luis; Jiménez, Carlos; Rodríguez, Jaime; Areche, Carlos; Chávez, Renato; Henríquez, Marlene; de la Cruz, Mercedes; Díaz, Caridad; Segade, Yuri; Vaca, Inmaculada

    2015-04-24

    Four new nitroasterric acid derivatives, pseudogymnoascins A-C (1-3) and 3-nitroasterric acid (4), along with the two known compounds questin and pyriculamide, were obtained from the cultures of a Pseudogymnoascus sp. fungus isolated from an Antarctic marine sponge belonging to the genus Hymeniacidon. The structures of the new compounds were determined by extensive NMR and MS analyses. These compounds are the first nitro derivatives of the known fungal metabolite asterric acid. Several asterric acid derivatives isolated from other fungal strains have shown antibacterial and antifungal activities. However, the new compounds described in this work were inactive against a panel of bacteria and fungi (MIC > 64 μg/mL).

  13. 12 CFR 563.172 - Financial derivatives.

    Code of Federal Regulations, 2011 CFR

    2011-01-01

    ... 12 Banks and Banking 5 2011-01-01 2011-01-01 false Financial derivatives. 563.172 Section 563.172...-OPERATIONS Financial Management Policies § 563.172 Financial derivatives. (a) What is a financial derivative? A financial derivative is a financial contract whose value depends on the value of one or more...

  14. The Dangers of Derivatives.

    ERIC Educational Resources Information Center

    Blumenstyk, Goldie

    1994-01-01

    Heavy investment in derivatives, whose value derives from the price movement of some underlying security, has meant heavy losses for some colleges and universities despite the fact that institutional investors are required to assure that they understand the risks of such investment. (MSE)

  15. A Photosensitizer-Loaded DNA Origami Nanosystem for Photodynamic Therapy.

    PubMed

    Zhuang, Xiaoxi; Ma, Xiaowei; Xue, Xiangdong; Jiang, Qiao; Song, Linlin; Dai, Luru; Zhang, Chunqiu; Jin, Shubin; Yang, Keni; Ding, Baoquan; Wang, Paul C; Liang, Xing-Jie

    2016-03-22

    Photodynamic therapy (PDT) offers an alternative for cancer treatment by using ultraviolet or visible light in the presence of a photosensitizer and molecular oxygen, which can produce highly reactive oxygen species that ultimately leading to the ablation of tumor cells by multifactorial mechanisms. However, this technique is limited by the penetration depth of incident light, the hypoxic environment of solid tumors, and the vulnerability of photobleaching reduces the efficiency of many imaging agents. In this work, we reported a cellular level dual-functional imaging and PDT nanosystem BMEPC-loaded DNA origami for photodynamic therapy with high efficiency and stable photoreactive property. The carbazole derivative BMEPC is a one- and two-photon imaging agent and photosensitizer with large two-photon absorption cross section, which can be fully excited by near-infrared light, and is also capable of destroying targets under anaerobic condition by generating reactive intermediates of Type I photodynamic reactions. However, the application of BMEPC was restricted by its poor solubility in aqueous environment and its aggregation caused quenching. We observed BMEPC-loaded DNA origami effectively reduced the photobleaching of BMEPC within cells. Upon binding to DNA origami, the intramolecular rotation of BMEPC became proper restricted, which intensify fluorescence emission and radicals production when being excited. After the BMEPC-loaded DNA origami are taken up by tumor cells, upon irradiation, BMEPC could generate free radicals and be released due to DNA photocleavage as well as the following partially degradation. Apoptosis was then induced by the generation of free radicals. This functional nanosystem provides an insight into the design of photosensitizer-loaded DNA origami for effective intracellular imaging and photodynamic therapy.

  16. Light and life in Baltimore--and beyond.

    PubMed

    Edidin, Michael

    2015-02-03

    Baltimore has been the home of numerous biophysical studies using light to probe cells. One such study, quantitative measurement of lateral diffusion of rhodopsin, set the standard for experiments in which recovery after photobleaching is used to measure lateral diffusion. Development of this method from specialized microscopes to commercial scanning confocal microscopes has led to widespread use of the technique to measure lateral diffusion of membrane proteins and lipids, and as well diffusion and binding interactions in cell organelles and cytoplasm. Perturbation of equilibrium distributions by photobleaching has also been developed into a robust method to image molecular proximity in terms of fluorescence resonance energy transfer between donor and acceptor fluorophores. Copyright © 2015 Biophysical Society. Published by Elsevier Inc. All rights reserved.

  17. Conformable derivative approach to anomalous diffusion

    NASA Astrophysics Data System (ADS)

    Zhou, H. W.; Yang, S.; Zhang, S. Q.

    2018-02-01

    By using a new derivative with fractional order, referred to conformable derivative, an alternative representation of the diffusion equation is proposed to improve the modeling of anomalous diffusion. The analytical solutions of the conformable derivative model in terms of Gauss kernel and Error function are presented. The power law of the mean square displacement for the conformable diffusion model is studied invoking the time-dependent Gauss kernel. The parameters related to the conformable derivative model are determined by Levenberg-Marquardt method on the basis of the experimental data of chloride ions transportation in reinforced concrete. The data fitting results showed that the conformable derivative model agrees better with the experimental data than the normal diffusion equation. Furthermore, the potential application of the proposed conformable derivative model of water flow in low-permeability media is discussed.

  18. Ferroelectricity in corundum derivatives

    NASA Astrophysics Data System (ADS)

    Ye, Meng; Vanderbilt, David

    The search for new ferroelectric (FE) materials holds promise for broadening our understanding of FE mechanisms and extending the range of application of FE materials. The known FE materials LiNbO3 can be regarded as derived from the A2O3 corundum structure with cation ordering. Here we consider more general binary (AB O3) and ternary (A2 BB' O6) corundum derivatives as an extended class of potential FE materials, motivated by the fact that some members of this class have recently been synthesized. There are four structure types for these corundum derivatives, and the number of cation combinations is enormous, but in many cases the energy barriers for polarization reversal may be too large to allow FE behavior. Here we present a first-principles study of the polar structure, coherent FE barrier, and domain-wall switching barrier for a representative set of polar corundum derivatives, allowing us to identify several potentially new FE materials. We also discuss the conditions under which ferroelectricity is compatible with magnetic ordering. Finally, we identify several empirical measures that can provide a rule of thumb for estimating the barrier energies. Our results should assist in the experimental search for new FE materials in the corundum derivative family. This work is supported by ONR Grant No. N-00014-12-1-1035.

  19. Asymptotic aspect of derivations in Banach algebras.

    PubMed

    Roh, Jaiok; Chang, Ick-Soon

    2017-01-01

    We prove that every approximate linear left derivation on a semisimple Banach algebra is continuous. Also, we consider linear derivations on Banach algebras and we first study the conditions for a linear derivation on a Banach algebra. Then we examine the functional inequalities related to a linear derivation and their stability. We finally take central linear derivations with radical ranges on semiprime Banach algebras and a continuous linear generalized left derivation on a semisimple Banach algebra.

  20. Evaluation of porphyrin C analogues for photodynamic therapy of cerebral glioma.

    PubMed

    Karagianis, G; Hill, J S; Stylli, S S; Kaye, A H; Varadaxis, N J; Reiss, J A; Phillips, D R

    1996-02-01

    A series of monomeric porphyrins (2-8) based on porphyrin C (1) have been tested as sensitisers for photodynamic therapy (PDT) of cerebral glioma using the in vitro/in vivo C6 intracerebral animal tumour model. The in vivo screening, consisting of cytotoxicity, phototoxicity (red light) and subcellular localisation studies, revealed two sensitisers (porphyrin 7, molecular weight 863 Da and porphyrin 8, molecular weight 889 Da), which had greater photoactivity than porphyrin C and similar photoactivity to haematoporphyrin derivative (HpD) although at a 5-fold higher dose than HpD. Both sensitisers showed intracellular localisation to discrete organelle sites and exhibited considerably less 'dark' cytotoxicity than HpD. The kinetics of uptake of porphyrins 7 and 8 was studied in the mouse C6 glioma model as well as in biopsy samples from normal brain, liver, spleen and blood. Maximal drug uptake levels in tumour occurred 9 and 6 h after intraperitoneal injection for 7 and 8 respectively, at which time the tumour to normal brain ratios were 15:1 and 13:1 respectively. The effect of PDT using porphyrin 7 activated by the gold metal vapour laser tuned to 627.8 nm was studied in Wistar rats bearing intracerebral C6 glioma. At a drug dose of 10 mg porphyrin 7 kg-1 body weight and laser doses of up to 400 J cm-2 light, selective tumour kill with sparing of normal brain was achieved, with a maximal depth of tumour kill of 1.77+/-0.40. mm. Irradiation following a higher drug dose of 75 mg porphyrin 7 kg-1 body weight resulted in a greater depth of tumour kill, but also significantly increased the likelihood and extent of necrosis in normal brain.

  1. Stability-Derivative Determination from Flight Data

    NASA Technical Reports Server (NTRS)

    Holowicz, Chester H.; Holleman, Euclid C.

    1958-01-01

    A comprehensive discussion of the various factors affecting the determination of stability and control derivatives from flight data is presented based on the experience of the NASA High-Speed Flight Station. Factors relating to test techniques, determination of mass characteristics, instrumentation, and methods of analysis are discussed. For most longitudinal-stability-derivative analyses simple equations utilizing period and damping have been found to be as satisfactory as more comprehensive methods. The graphical time-vector method has been the basis of lateral-derivative analysis, although simple approximate methods can be useful If applied with caution. Control effectiveness has been generally obtained by relating the peak acceleration to the rapid control input, and consideration must be given to aerodynamic contributions if reasonable accuracy is to be realized.. Because of the many factors involved In the determination of stability derivatives, It is believed that the primary stability and control derivatives are probably accurate to within 10 to 25 percent, depending upon the specific derivative. Static-stability derivatives at low angle of attack show the greatest accuracy.

  2. 5 CFR 1312.7 - Derivative classification.

    Code of Federal Regulations, 2010 CFR

    2010-01-01

    ..., DOWNGRADING, DECLASSIFICATION AND SAFEGUARDING OF NATIONAL SECURITY INFORMATION Classification and Declassification of National Security Information § 1312.7 Derivative classification. A derivative classification... 5 Administrative Personnel 3 2010-01-01 2010-01-01 false Derivative classification. 1312.7 Section...

  3. Optimal economic order quantity for buyer-distributor-vendor supply chain with backlogging derived without derivatives

    NASA Astrophysics Data System (ADS)

    Teng, Jinn-Tsair; Cárdenas-Barrón, Leopoldo Eduardo; Lou, Kuo-Ren; Wee, Hui Ming

    2013-05-01

    In this article, we first complement an inappropriate mathematical error on the total cost in the previously published paper by Chung and Wee [2007, 'Optimal the Economic Lot Size of a Three-stage Supply Chain With Backlogging Derived Without Derivatives', European Journal of Operational Research, 183, 933-943] related to buyer-distributor-vendor three-stage supply chain with backlogging derived without derivatives. Then, an arithmetic-geometric inequality method is proposed not only to simplify the algebraic method of completing prefect squares, but also to complement their shortcomings. In addition, we provide a closed-form solution to integral number of deliveries for the distributor and the vendor without using complex derivatives. Furthermore, our method can solve many cases in which their method cannot, because they did not consider that a squared root of a negative number does not exist. Finally, we use some numerical examples to show that our proposed optimal solution is cheaper to operate than theirs.

  4. 28 CFR 17.26 - Derivative classification.

    Code of Federal Regulations, 2014 CFR

    2014-07-01

    ... 28 Judicial Administration 1 2014-07-01 2014-07-01 false Derivative classification. 17.26 Section... ACCESS TO CLASSIFIED INFORMATION Classified Information § 17.26 Derivative classification. (a) Persons need not possess original classification authority to derivatively classify information based on source...

  5. 28 CFR 17.26 - Derivative classification.

    Code of Federal Regulations, 2013 CFR

    2013-07-01

    ... 28 Judicial Administration 1 2013-07-01 2013-07-01 false Derivative classification. 17.26 Section... ACCESS TO CLASSIFIED INFORMATION Classified Information § 17.26 Derivative classification. (a) Persons need not possess original classification authority to derivatively classify information based on source...

  6. 28 CFR 17.26 - Derivative classification.

    Code of Federal Regulations, 2012 CFR

    2012-07-01

    ... 28 Judicial Administration 1 2012-07-01 2012-07-01 false Derivative classification. 17.26 Section... ACCESS TO CLASSIFIED INFORMATION Classified Information § 17.26 Derivative classification. (a) Persons need not possess original classification authority to derivatively classify information based on source...

  7. 22 CFR 9.6 - Derivative classification.

    Code of Federal Regulations, 2010 CFR

    2010-04-01

    ... 22 Foreign Relations 1 2010-04-01 2010-04-01 false Derivative classification. 9.6 Section 9.6 Foreign Relations DEPARTMENT OF STATE GENERAL SECURITY INFORMATION REGULATIONS § 9.6 Derivative classification. (a) Definition. Derivative classification is the incorporating, paraphrasing, restating or...

  8. 28 CFR 17.26 - Derivative classification.

    Code of Federal Regulations, 2011 CFR

    2011-07-01

    ... 28 Judicial Administration 1 2011-07-01 2011-07-01 false Derivative classification. 17.26 Section... ACCESS TO CLASSIFIED INFORMATION Classified Information § 17.26 Derivative classification. (a) Persons need not possess original classification authority to derivatively classify information based on source...

  9. 6 CFR 7.26 - Derivative classification.

    Code of Federal Regulations, 2012 CFR

    2012-01-01

    ... 6 Domestic Security 1 2012-01-01 2012-01-01 false Derivative classification. 7.26 Section 7.26... INFORMATION Classified Information § 7.26 Derivative classification. (a) Derivative classification is defined... already classified, and marking the newly developed material consistent with the classification markings...

  10. 32 CFR 2001.22 - Derivative classification.

    Code of Federal Regulations, 2012 CFR

    2012-07-01

    ... 32 National Defense 6 2012-07-01 2012-07-01 false Derivative classification. 2001.22 Section 2001... Identification and Markings § 2001.22 Derivative classification. (a) General. Information classified derivatively on the basis of source documents or classification guides shall bear all markings prescribed in...

  11. 32 CFR 2001.22 - Derivative classification.

    Code of Federal Regulations, 2013 CFR

    2013-07-01

    ... 32 National Defense 6 2013-07-01 2013-07-01 false Derivative classification. 2001.22 Section 2001... Identification and Markings § 2001.22 Derivative classification. (a) General. Information classified derivatively on the basis of source documents or classification guides shall bear all markings prescribed in...

  12. 6 CFR 7.26 - Derivative classification.

    Code of Federal Regulations, 2013 CFR

    2013-01-01

    ... 6 Domestic Security 1 2013-01-01 2013-01-01 false Derivative classification. 7.26 Section 7.26... INFORMATION Classified Information § 7.26 Derivative classification. (a) Derivative classification is defined... already classified, and marking the newly developed material consistent with the classification markings...

  13. 6 CFR 7.26 - Derivative classification.

    Code of Federal Regulations, 2014 CFR

    2014-01-01

    ... 6 Domestic Security 1 2014-01-01 2014-01-01 false Derivative classification. 7.26 Section 7.26... INFORMATION Classified Information § 7.26 Derivative classification. (a) Derivative classification is defined... already classified, and marking the newly developed material consistent with the classification markings...

  14. 32 CFR 2001.22 - Derivative classification.

    Code of Federal Regulations, 2014 CFR

    2014-07-01

    ... 32 National Defense 6 2014-07-01 2014-07-01 false Derivative classification. 2001.22 Section 2001... Identification and Markings § 2001.22 Derivative classification. (a) General. Information classified derivatively on the basis of source documents or classification guides shall bear all markings prescribed in...

  15. 6 CFR 7.26 - Derivative classification.

    Code of Federal Regulations, 2010 CFR

    2010-01-01

    ... 6 Domestic Security 1 2010-01-01 2010-01-01 false Derivative classification. 7.26 Section 7.26 Domestic Security DEPARTMENT OF HOMELAND SECURITY, OFFICE OF THE SECRETARY CLASSIFIED NATIONAL SECURITY INFORMATION Classified Information § 7.26 Derivative classification. (a) Derivative classification is defined...

  16. 32 CFR 2001.22 - Derivative classification.

    Code of Federal Regulations, 2011 CFR

    2011-07-01

    ... 32 National Defense 6 2011-07-01 2011-07-01 false Derivative classification. 2001.22 Section 2001... Identification and Markings § 2001.22 Derivative classification. (a) General. Information classified derivatively on the basis of source documents or classification guides shall bear all markings prescribed in...

  17. Handheld dual fluorescence and reflection spectroscopy system for monitoring topical low dose ALA-PDT of actinic keratoses (AK)

    NASA Astrophysics Data System (ADS)

    Charamisinau, Ivan; Keymel, Kenneth; Potter, William; Oseroff, Allan R.

    2006-02-01

    Photodynamic therapy is an effective, minimally invasive skin cancer treatment modality with few side effects. Improved therapeutic selectivity and efficacy is expected if treatment is optimized individually for each patient based on detailed measurements prior and during the treatment. The handheld system presented allows measuring optical properties of the skin, the rate of photosensitizer photobleaching during the ALA PDT and oxygen saturation in the tissue. The photobleaching rate is monitored using fluorescence spectroscopy, where protoporphyrin IX in tissue is exited by 410 nm (blue) or 532 nm (green) laser light, and fluorescence in the 580-800 nm range is monitored. The photobleaching rate is calculated by correlating the measured spectrum with known protoporphyrin IX, photoproduct and nonspecific tissue autofluorescence spectra using correlation analysis. Double-wavelength excitation allows a rough estimation of the depth of the fluorescence source due to the significant difference in penetration depth for blue and green light. Blood concentration and oxygenation in the tissue are found from the white light reflectance spectrum in the 460-800 nm range. Known spectra for the oxy- and deoxyhemoglobin, melanin, and tissue baseline absorption and tissue scattering are substituted in nonlinear equations to find the penetration depth and diffuse reflectance coefficient. The nonlinear equation for the diffuse reflectance coefficient is solved for blood and melanin concentrations and blood oxygenation values that provide the best fit to the measured spectrum. The optical properties of the tissue obtained from the reflectance spectroscopy are used to correct the fluorescence data. A noncontact probe with 5 fibers (3 excitation and 2 detection) focused to the same 5 mm diameter spot: 2 excitation lasers, a white light lamp and a two-channel spectrometer are used. A LabView program with custom nonlinear equation solvers written in C++ automatically performs the

  18. 12 CFR 403.4 - Derivative classification.

    Code of Federal Regulations, 2014 CFR

    2014-01-01

    ... 12 Banks and Banking 5 2014-01-01 2014-01-01 false Derivative classification. 403.4 Section 403.4 Banks and Banking EXPORT-IMPORT BANK OF THE UNITED STATES CLASSIFICATION, DECLASSIFICATION, AND SAFEGUARDING OF NATIONAL SECURITY INFORMATION § 403.4 Derivative classification. (a) Use of derivative...

  19. 12 CFR 403.4 - Derivative classification.

    Code of Federal Regulations, 2013 CFR

    2013-01-01

    ... 12 Banks and Banking 5 2013-01-01 2013-01-01 false Derivative classification. 403.4 Section 403.4 Banks and Banking EXPORT-IMPORT BANK OF THE UNITED STATES CLASSIFICATION, DECLASSIFICATION, AND SAFEGUARDING OF NATIONAL SECURITY INFORMATION § 403.4 Derivative classification. (a) Use of derivative...

  20. 12 CFR 403.4 - Derivative classification.

    Code of Federal Regulations, 2012 CFR

    2012-01-01

    ... 12 Banks and Banking 5 2012-01-01 2012-01-01 false Derivative classification. 403.4 Section 403.4 Banks and Banking EXPORT-IMPORT BANK OF THE UNITED STATES CLASSIFICATION, DECLASSIFICATION, AND SAFEGUARDING OF NATIONAL SECURITY INFORMATION § 403.4 Derivative classification. (a) Use of derivative...

  1. 40 CFR 721.5913 - Phenothiazine derivative.

    Code of Federal Regulations, 2011 CFR

    2011-07-01

    ... 40 Protection of Environment 31 2011-07-01 2011-07-01 false Phenothiazine derivative. 721.5913... Substances § 721.5913 Phenothiazine derivative. (a) Chemical substance and significant new uses subject to reporting. (1) The chemical substance identified generically as a phenothiazine derivative (PMN P-96-813) is...

  2. 40 CFR 721.5913 - Phenothiazine derivative.

    Code of Federal Regulations, 2010 CFR

    2010-07-01

    ... 40 Protection of Environment 30 2010-07-01 2010-07-01 false Phenothiazine derivative. 721.5913... Substances § 721.5913 Phenothiazine derivative. (a) Chemical substance and significant new uses subject to reporting. (1) The chemical substance identified generically as a phenothiazine derivative (PMN P-96-813) is...

  3. 12 CFR 403.4 - Derivative classification.

    Code of Federal Regulations, 2011 CFR

    2011-01-01

    ... 12 Banks and Banking 4 2011-01-01 2011-01-01 false Derivative classification. 403.4 Section 403.4 Banks and Banking EXPORT-IMPORT BANK OF THE UNITED STATES CLASSIFICATION, DECLASSIFICATION, AND SAFEGUARDING OF NATIONAL SECURITY INFORMATION § 403.4 Derivative classification. (a) Use of derivative...

  4. 12 CFR 563.172 - Financial derivatives.

    Code of Federal Regulations, 2010 CFR

    2010-01-01

    ... underlying assets, indices, or reference rates. The most common types of financial derivatives are futures, forward commitments, options, and swaps. A mortgage derivative security, such as a collateralized mortgage... 12 Banks and Banking 5 2010-01-01 2010-01-01 false Financial derivatives. 563.172 Section 563.172...

  5. 40 CFR 721.1820 - Bisphenol derivative.

    Code of Federal Regulations, 2010 CFR

    2010-07-01

    ... 40 Protection of Environment 30 2010-07-01 2010-07-01 false Bisphenol derivative. 721.1820 Section... Substances § 721.1820 Bisphenol derivative. (a) Chemical substance and significant new uses subject to reporting. (1) The chemical substance identified generically as bisphenol derivative (PMN No. P-92-509) is...

  6. 40 CFR 721.9658 - Thiadiazole derivative.

    Code of Federal Regulations, 2010 CFR

    2010-07-01

    ... 40 Protection of Environment 30 2010-07-01 2010-07-01 false Thiadiazole derivative. 721.9658... Substances § 721.9658 Thiadiazole derivative. (a) Chemical substance and significant new uses subject to reporting. (1) The chemical substance identified as a thiadiazole derivative (PMN P-94-1631) is subject to...

  7. 40 CFR 721.9658 - Thiadiazole derivative.

    Code of Federal Regulations, 2011 CFR

    2011-07-01

    ... 40 Protection of Environment 31 2011-07-01 2011-07-01 false Thiadiazole derivative. 721.9658... Substances § 721.9658 Thiadiazole derivative. (a) Chemical substance and significant new uses subject to reporting. (1) The chemical substance identified as a thiadiazole derivative (PMN P-94-1631) is subject to...

  8. 40 CFR 721.1820 - Bisphenol derivative.

    Code of Federal Regulations, 2011 CFR

    2011-07-01

    ... 40 Protection of Environment 31 2011-07-01 2011-07-01 false Bisphenol derivative. 721.1820 Section... Substances § 721.1820 Bisphenol derivative. (a) Chemical substance and significant new uses subject to reporting. (1) The chemical substance identified generically as bisphenol derivative (PMN No. P-92-509) is...

  9. On the ψ-Hilfer fractional derivative

    NASA Astrophysics Data System (ADS)

    Vanterler da C. Sousa, J.; Capelas de Oliveira, E.

    2018-07-01

    In this paper we introduce a new fractional derivative with respect to another function the so-called ψ-Hilfer fractional derivative. We discuss some properties and important results of the fractional calculus. In this sense, we present some results involving uniformly convergent sequence of function, uniformly continuous function and examples including the Mittag-Leffler function with one parameter. Finally, we present a wide class of integrals and fractional derivatives, by means of the fractional integral with respect to another function and the ψ-Hilfer fractional derivative.

  10. Statistical inference in single molecule measurements of protein adsorption

    NASA Astrophysics Data System (ADS)

    Armstrong, Megan J.; Tsitkov, Stanislav; Hess, Henry

    2018-02-01

    Significant effort has been invested into understanding the dynamics of protein adsorption on surfaces, in particular to predict protein behavior at the specialized surfaces of biomedical technologies like hydrogels, nanoparticles, and biosensors. Recently, the application of fluorescent single molecule imaging to this field has permitted the tracking of individual proteins and their stochastic contribution to the aggregate dynamics of adsorption. However, the interpretation of these results is complicated by (1) the finite time available to observe effectively infinite adsorption timescales and (2) the contribution of photobleaching kinetics to adsorption kinetics. Here, we perform a protein adsorption simulation to introduce specific survival analysis methods that overcome the first complication. Additionally, we collect single molecule residence time data from the adsorption of fibrinogen to glass and use survival analysis to distinguish photobleaching kinetics from protein adsorption kinetics.

  11. Quinazoline derivatives: synthesis and bioactivities

    PubMed Central

    2013-01-01

    Owing to the significant biological activities, quinazoline derivatives have drawn more and more attention in the synthesis and bioactivities research. This review summarizes the recent advances in the synthesis and biological activities investigations of quinazoline derivatives. According to the main method the authors adopted in their research design, those synthetic methods were divided into five main classifications, including Aza-reaction, Microwave-assisted reaction, Metal-mediated reaction, Ultrasound-promoted reaction and Phase-transfer catalysis reaction. The biological activities of the synthesized quinazoline derivatives also are discussed. PMID:23731671

  12. Live-cell imaging of phosphatidic acid dynamics in pollen tubes visualized by Spo20p-derived biosensor.

    PubMed

    Potocký, Martin; Pleskot, Roman; Pejchar, Přemysl; Vitale, Nicolas; Kost, Benedikt; Zárský, Viktor

    2014-07-01

    Although phosphatidic acid (PA) is structurally the simplest membrane phospholipid, it has been implicated in the regulation of many cellular events, including cytoskeletal dynamics, membrane trafficking and stress responses. Plant PA shows rapid turnover but the information about its spatio-temporal distribution in plant cells is missing. Here we demonstrate the use of a lipid biosensor that enables us to monitor PA dynamics in plant cells. The biosensor consists of a PA-binding domain of yeast SNARE Spo20p fused to fluorescent proteins. Live-cell imaging of PA dynamics in transiently transformed tobacco (Nicotiana tabacum) pollen tubes was performed using confocal laser scanning microscopy. In growing pollen tubes, PA shows distinct annulus-like fluorescence pattern in the plasma membrane behind the extreme tip. Coexpression studies with markers for other plasmalemma signaling lipids phosphatidylinositol 4,5-bisphosphate and diacylglycerol revealed limited colocalization at the shoulders of the apex. PA distribution and concentrations show distinct responses to various lipid signaling inhibitors. Fluorescence recovery after photobleaching (FRAP) analysis suggests high PA turnover in the plasma membrane. Our data show that a biosensor based on the Spo20p-PA binding domain is suitable for live-cell imaging of PA also in plant cells. In tobacco pollen tubes, distinct subapical PA maximum corroborates its involvement in the regulation of endocytosis and actin dynamics. © 2014 The Authors. New Phytologist © 2014 New Phytologist Trust.

  13. Risk management, derivatives and shariah compliance

    NASA Astrophysics Data System (ADS)

    Bacha, Obiyathulla Ismath

    2013-04-01

    Despite the impressive growth of Islamic Banking and Finance (IBF), a number of weaknesses remain. The most important of this is perhaps the lack of shariah compliant risk management tools. While the risk sharing philosophy of Islamic Finance requires the acceptance of risk to justify returns, the shariah also requires adherents to avoid unnecessary risk-maysir. The requirement to avoid maysir is in essence a call for the prudent management of risk. Contemporary risk management revolves around financial engineering, the building blocks of which are financial derivatives. Despite the proven efficacy of derivatives in the management of risk in the conventional space, shariah scholars appear to be suspicious and uneasy with their use in IBF. Some have imposed outright prohibition of their use. This paper re-examines the issue of contemporary derivative instruments and shariah compliance. The shariah compatibility of derivatives is shown in a number of ways. First, by way of qualitative evaluation of whether derivatives can be made to comply with the key prohibitions of the sharia. Second, by way of comparing the payoff profiles of derivatives with risk sharing finance and Bai Salam contracts. Finally, the equivalence between shariah compliant derivatives like the IPRS and Islamic FX Currency Forwards with conventional ones is presented.

  14. Bioactive terpenes from marine-derived fungi.

    PubMed

    Elissawy, Ahmed M; El-Shazly, Mohamed; Ebada, Sherif S; Singab, AbdelNasser B; Proksch, Peter

    2015-04-03

    Marine-derived fungi continue to be a prolific source of secondary metabolites showing diverse bioactivities. Terpenoids from marine-derived fungi exhibit wide structural diversity including numerous compounds with pronounced biological activities. In this review, we survey the last five years' reports on terpenoidal metabolites from marine-derived fungi with particular attention on those showing marked biological activities.

  15. Generalized Fractional Derivative Anisotropic Viscoelastic Characterization.

    PubMed

    Hilton, Harry H

    2012-01-18

    Isotropic linear and nonlinear fractional derivative constitutive relations are formulated and examined in terms of many parameter generalized Kelvin models and are analytically extended to cover general anisotropic homogeneous or non-homogeneous as well as functionally graded viscoelastic material behavior. Equivalent integral constitutive relations, which are computationally more powerful, are derived from fractional differential ones and the associated anisotropic temperature-moisture-degree-of-cure shift functions and reduced times are established. Approximate Fourier transform inversions for fractional derivative relations are formulated and their accuracy is evaluated. The efficacy of integer and fractional derivative constitutive relations is compared and the preferential use of either characterization in analyzing isotropic and anisotropic real materials must be examined on a case-by-case basis. Approximate protocols for curve fitting analytical fractional derivative results to experimental data are formulated and evaluated.

  16. Controllable Syntheses of MOF-Derived Materials.

    PubMed

    Zou, Kang-Yu; Li, Zuo-Xi

    2018-05-02

    Metal-organic frameworks (MOFs), as an important kind of porous inorganic-organic hybrid materials with inherent outstanding physicochemistry characteristics, can be widely applied as versatile precursors for the facile preparation of functional MOF-derived materials. However, there are plenty of sophisticated factors during the synthetic process, which is far from reaching the goal of effectively controlling the nature of MOF-derived materials (such as the composition, morphology and surface area). Therefore, it is urgently necessary to develop regular protocols and concepts for controllable syntheses of MOF-derived materials. In this minireview, we mainly summarize and analyze complicated factors in the fabrication of MOF-derived materials according to recently reported literatures, and this provides a new insight into the rational design and syntheses of MOF-derived materials. © 2018 Wiley-VCH Verlag GmbH & Co. KGaA, Weinheim.

  17. Ferroelectricity in corundum derivatives

    NASA Astrophysics Data System (ADS)

    Ye, Meng; Vanderbilt, David

    2016-04-01

    The search for new ferroelectric (FE) materials holds promise for broadening our understanding of FE mechanisms and extending the range of application of FE materials. Here we investigate a class of A B O3 and A2B B'O6 materials that can be derived from the X2O3 corundum structure by mixing two or three ordered cations on the X site. Most such corundum derivatives have a polar structure, but it is unclear whether the polarization is reversible, which is a requirement for a FE material. In this paper, we propose a method to study the FE reversal path of materials in the corundum derivative family. We first categorize the corundum derivatives into four classes and show that only two of these allow for the possibility of FE reversal. We then calculate the energy profile and energy barrier of the FE reversal path using first-principles density functional methods with a structural constraint. Furthermore, we identify several empirical measures that can provide a rule of thumb for estimating the energy barriers. Finally, the conditions under which the magnetic ordering is compatible with ferroelectricity are determined. These results lead us to predict several potentially new FE materials.

  18. Photostable fluorescent organic dots with aggregation-induced emission (AIE dots) for noninvasive long-term cell tracing

    NASA Astrophysics Data System (ADS)

    Li, Kai; Qin, Wei; Ding, Dan; Tomczak, Nikodem; Geng, Junlong; Liu, Rongrong; Liu, Jianzhao; Zhang, Xinhai; Liu, Hongwei; Liu, Bin; Tang, Ben Zhong

    2013-01-01

    Long-term noninvasive cell tracing by fluorescent probes is of great importance to life science and biomedical engineering. For example, understanding genesis, development, invasion and metastasis of cancerous cells and monitoring tissue regeneration after stem cell transplantation require continual tracing of the biological processes by cytocompatible fluorescent probes over a long period of time. In this work, we successfully developed organic far-red/near-infrared dots with aggregation-induced emission (AIE dots) and demonstrated their utilities as long-term cell trackers. The high emission efficiency, large absorptivity, excellent biocompatibility, and strong photobleaching resistance of the AIE dots functionalized by cell penetrating peptides derived from transactivator of transcription proteins ensured outstanding long-term noninvasive in vitro and in vivo cell tracing. The organic AIE dots outperform their counterparts of inorganic quantum dots, opening a new avenue in the development of fluorescent probes for following biological processes such as carcinogenesis.

  19. Sol-gel derived sorbents

    DOEpatents

    Sigman, Michael E.; Dindal, Amy B.

    2003-11-11

    Described is a method for producing copolymerized sol-gel derived sorbent particles for the production of copolymerized sol-gel derived sorbent material. The method for producing copolymerized sol-gel derived sorbent particles comprises adding a basic solution to an aqueous metal alkoxide mixture for a pH.ltoreq.8 to hydrolyze the metal alkoxides. Then, allowing the mixture to react at room temperature for a precalculated period of time for the mixture to undergo an increased in viscosity to obtain a desired pore size and surface area. The copolymerized mixture is then added to an immiscible, nonpolar solvent that has been heated to a sufficient temperature wherein the copolymerized mixture forms a solid upon the addition. The solid is recovered from the mixture, and is ready for use in an active sampling trap or activated for use in a passive sampling trap.

  20. Synthesis and Cytotoxicities of Royleanone Derivatives.

    PubMed

    Li, Cheng-Ji; Xia, Fan; Wu, Rong; Tan, Hong-Sheng; Xu, Hong-Xi; Xu, Gang; Qin, Hong-Bo

    2018-06-16

    Carnosic acid was used as starting material to synthesize royleanone derivatives featured C11-C14 para quinone. The importance of C-20 group of royleanone derivatives was verified by the cytotoxicity assay of royleanonic acid, miltionone I and deoxyneocrptotanshinone. Following our synthetic route, 15 amide derivatives were synthesized and 8 compounds exhibited moderate cytotoxic activities against three human cancer lines in vitro.

  1. Relieving of Misconceptions of Derivative Concept with Derive

    ERIC Educational Resources Information Center

    Kaplan, Abdullah; Ozturk, Mesut; Ocal, Mehmet Fatih

    2015-01-01

    The purpose of this study is to determine students' learning levels in derivative subjects and their misconceptions. In addition, this study aims to compared to the effects of the computer based instruction and traditional instruction in resolving these misconceptions. For this purpose, 12th grade 70 students were chosen from high schools in Agri…

  2. Relieving of Misconceptions of Derivative Concept with Derive

    ERIC Educational Resources Information Center

    Kaplan, Abdullah; Ozturk, Mesut; Ocal, Mehmet Fatih

    2015-01-01

    The purpose of this study is to determine students' learning levels in derivative subjects and their misconceptions. In addition, this study aims to compared to the effects of the computer based instruction and traditional instruction in resolving these misconceptions. For this purpose, 70 12th grade students were chosen from high schools in Agri…

  3. A heterotrimer model of the complete Microprocessor complex revealed by single-molecule subunit counting.

    PubMed

    Herbert, Kristina M; Sarkar, Susanta K; Mills, Maria; Delgado De la Herran, Hilda C; Neuman, Keir C; Steitz, Joan A

    2016-02-01

    During microRNA (miRNA) biogenesis, the Microprocessor complex (MC), composed minimally of Drosha, an RNaseIII enzyme, and DGCR8, a double-stranded RNA-binding protein, cleaves the primary-miRNA (pri-miRNA) to release the pre-miRNA stem-loop structure. Size-exclusion chromatography of the MC, isolated from mammalian cells, suggested multiple copies of one or both proteins in the complex. However, the exact stoichiometry was unknown. Initial experiments suggested that DGCR8 bound pri-miRNA substrates specifically, and given that Drosha could not be bound or cross-linked to RNA, a sequential model for binding was established in which DGCR8 bound first and recruited Drosha. Therefore, many laboratories have studied DGCR8 binding to RNA in the absence of Drosha and have shown that deletion constructs of DGCR8 can multimerize in the presence of RNA. More recently, it was demonstrated that Drosha can bind pri-miRNA substrates in the absence of DGCR8, casting doubt on the sequential model of binding. In the same study, using a single-molecule photobleaching assay, fluorescent protein-tagged deletion constructs of DGCR8 and Drosha assembled into a heterotrimeric complex on RNA, comprising two DGCR8 molecules and one Drosha molecule. To determine the stoichiometry of Drosha and DGCR8 within the MC in the absence of added RNA, we also used a single-molecule photobleaching assay and confirmed the heterotrimeric model of the human MC. We demonstrate that a heterotrimeric complex is likely preformed in the absence of RNA and exists even when full-length proteins are expressed and purified from human cells, and when hAGT-derived tags are used rather than fluorescent proteins. © 2016 Herbert et al.; Published by Cold Spring Harbor Laboratory Press for the RNA Society.

  4. Derivation and characterization of a pig embryonic stem cell-derived exocrine pancreatic cell line

    USDA-ARS?s Scientific Manuscript database

    The establishment and initial characterization of a pig embryonic stem cell-derived pancreatic cell line, PICM-31, and a colony-cloned derivative cell line, PICM-31A, is described. The cell lines were propagated for several months at split ratios of 1:3 or 1:5 at each passage on STO feeder cells af...

  5. Photobleachable Diazonium Salt-Phenolic Resin Two-Layer Resist System

    NASA Astrophysics Data System (ADS)

    Uchino, Shou-ichi; Iwayanagi, Takao; Hashimoto, Michiaki

    1988-01-01

    This article describes a new negative two-layer photoresist system formed by a simple, successive spin-coating method. An aqueous acetic acid solution of diazonium salt and poly(N-vinylpyrrolidone) is deposited so as to contact a phenolic resin film spin-coated on a silicon wafer. The diazonium salt diffuses into the phenolic resin layer after standing for several minutes. The residual solution on the phenolic resin film doped with diazonium salt is spun to form the diazonium salt-poly(N-vinylpyrrolidone) top layer. This forms a uniform two-layer resist without phase separation or striation. Upon UV exposure, the diazonium salt in the top layer bleaches to act as a CEL dye, while the diazonium salt in the bottom layer decomposes to cause insolubilization. Half μm line-and-space patterns are obtained with an i-line stepper using 4-diazo-N,N-dimethylaniline chloride zinc chloride double salt as the diazonium salt and a cresol novolac resin for the bottom polymer layer. The resist formation processes, insolubilization mechanism, and the resolution capability of the new two-layer resist are discussed.

  6. Perturbative reduction of derivative order in EFT

    NASA Astrophysics Data System (ADS)

    Glavan, Dražen

    2018-02-01

    Higher derivative corrections are ubiquitous in effective field theories, which seemingly introduces new degrees of freedom at successive orders. This is actually an artefact of the implicit local derivative expansion defining effective field theories. We argue that higher derivative corrections that introduce additional degrees of freedom should be removed and their effects captured either by lower derivative corrections, or special combinations of higher derivative corrections not propagating extra degrees of freedom. Three methods adapted for this task are examined and field redefinitions are found to be most appropriate. First order higher derivative corrections in a scalar tensor theory are removed by field redefinition and it is found that their effects are captured by a subset of Horndeski theories. A case is made for restricting the effective field theory expansions in principle to only terms not introducing additional degrees of freedom.

  7. Using derivatives to hedge against the unexpected.

    PubMed

    Aderholt, J M; Rasmussen, R H

    1996-02-01

    Derivatives--financial instruments with a rate of return derived from an underlying asset--have been used as investment instruments for decades. Many derivative products originally were created explicitly for the purpose of reducing financial risks and have become more widely used and more complex in recent years. Healthcare financial managers should have a basic understanding of derivatives as well as the ability to apply general guidelines for their appropriate use in healthcare financial management.

  8. Shuttling of G protein subunits between the plasma membrane and intracellular membranes.

    PubMed

    Chisari, Mariangela; Saini, Deepak Kumar; Kalyanaraman, Vani; Gautam, Narasimhan

    2007-08-17

    Heterotrimeric G proteins (alphabetagamma) mediate the majority of signaling pathways in mammalian cells. It is long held that G protein function is localized to the plasma membrane. Here we examined the spatiotemporal dynamics of G protein localization using fluorescence recovery after photobleaching, fluorescence loss in photobleaching, and a photoswitchable fluorescent protein, Dronpa. Unexpectedly, G protein subunits shuttle rapidly (t1/2 < 1 min) between the plasma membrane and intracellular membranes. We show that consistent with such shuttling, G proteins constitutively reside in endomembranes. Furthermore, we show that shuttling is inhibited by 2-bromopalmitate. Thus, contrary to present thought, G proteins do not reside permanently on the plasma membrane but are constantly testing the cytoplasmic surfaces of the plasma membrane and endomembranes to maintain G protein pools in intracellular membranes to establish direct communication between receptors and endomembranes.

  9. Human induced pluripotent stem cell (hiPSC)-derived neurons respond to convulsant drugs when co-cultured with hiPSC-derived astrocytes.

    PubMed

    Ishii, Misawa Niki; Yamamoto, Koji; Shoji, Masanobu; Asami, Asano; Kawamata, Yuji

    2017-08-15

    Accurate risk assessment for drug-induced seizure is expected to be performed before entering clinical studies because of its severity and fatal damage to drug development. Induced pluripotent stem cell (iPSC) technology has allowed the use of human neurons and glial cells in toxicology studies. Recently, several studies showed the advantage of co-culture system of human iPSC (hiPSC)-derived neurons with rodent/human primary astrocytes regarding neuronal functions. However, the application of hiPSC-derived neurons for seizure risk assessment has not yet been fully addressed, and not at all when co-cultured with hiPSC-derived astrocytes. Here, we characterized hiPSC-derived neurons co-cultured with hiPSC-derived astrocytes to discuss how hiPSC-derived neurons are useful to assess seizure risk of drugs. First, we detected the frequency of spikes and synchronized bursts hiPSC-derived neurons when co-cultured with hiPSC-derived astrocytes for 8 weeks. This synchronized burst was suppressed by the treatment with 6-cyano-7-nitroquinoxaline-2,3-dione, α-amino-3-hydroxy-5-methylisoxazole-4-propionic acid (AMPA) receptor antagonist, and D-(-)-2-amino-5-phosphonopentanoic acid, an N-Methyl-d-aspartate (NMDA) receptor antagonist. These data suggested that co-cultured hiPSC-derived neurons formed synaptic connections mediated by AMPA and NMDA receptors. We also demonstrated that co-cultured hiPSC-derived neurons showed epileptiform activity upon treatment with gabazine or kaliotoxin. Finally, we performed single-cell transcriptome analysis in hiPSC-derived neurons and found that hiPSC-derived astrocytes activated the pathways involved in the activities of AMPA and NMDA receptor functions, neuronal polarity, and axon guidance in hiPSC-derived neurons. These data suggested that hiPSC-derived astrocytes promoted the development of action potential, synaptic functions, and neuronal networks in hiPSC-derived neurons, and then these functional alterations result in the epileptiform

  10. Synthesis of octahedral like Cu-BTC derivatives derived from MOF calcined under different atmosphere for application in CO oxidation

    NASA Astrophysics Data System (ADS)

    Yang, Yiqiong; Dong, Han; Wang, Yin; He, Chi; Wang, Yuxin; Zhang, Xiaodong

    2018-02-01

    A series of octahedral structure Cu-BTC derivatives were successfully achieved through direct calcination of copper based metal organic framework Cu-BTC under different atmosphere (CO reaction gas, oxidizing gas O2, reducing gas H2, inert gas Ar). The Cu-BTC derivatives were characterized by X-ray diffraction (XRD), scanning electron microscope (SEM), transmission electron microscopy (TEM), high-resolution transmission electron microscopy (HRTEM), laser Raman spectroscopy (LRS), N2 adsorption-desorption isotherm, element analysis, H2-temperature program reduction (H2-TPR) and X-ray photoelectron spectroscopic (XPS). It is found that Cu-BTC derivative derived from MOF calcined under reaction gas/O2 (Cu-BTC-CO/Cu-BTC-O) only retain Cu2O and CuO species. In addition, a weak Cu-BTC structure and Cu particles were observed on Cu-BTC derivative derived from MOF calcined under H2 (Cu-BTC-H). Obviously differently, Cu-BTC derivative derived from MOF calcined under Ar (Cu-BTC-Ar) still retains good MOF structure. The catalytic performance for CO oxidation over Cu-BTC derivatives was studied. It was found that Cu-BTC-CO showed a smaller specific surface area (8.0 m2/g), but presented an excellent catalytic performance, long-term stability and cycling stability with a complete CO conversion temperature (T100) of 140 °C, which was ascribed to the higher Cu2O/CuO ratio, good low temperature reduction behavior and a high quantity of surface active oxygen species.

  11. Territrem and Butyrolactone Derivatives from a Marine-Derived Fungus Aspergillus Terreus

    PubMed Central

    Nong, Xu-Hua; Wang, Yi-Fei; Zhang, Xiao-Yong; Zhou, Mu-Ping; Xu, Xin-Ya; Qi, Shu-Hua

    2014-01-01

    Seventeen lactones including eight territrem derivatives (1–8) and nine butyrolactone derivatives (9–17) were isolated from a marine-derived fungus Aspergillus terreus SCSGAF0162 under solid-state fermentation of rice. Compounds 1–3 and 9–10 were new, and their structures were elucidated by spectroscopic analysis. The acetylcholinesterase inhibitory activity and antiviral activity of compounds 1–17 were evaluated. Among them, compounds 1 and 2 showed strong inhibitory activity against acetylcholinesterase with IC50 values of 4.2 ± 0.6, 4.5 ± 0.6 nM, respectively. This is the first time it has been reported that 3, 6, 10, 12 had evident antiviral activity towards HSV-1 with IC50 values of 16.4 ± 0.6, 6.34 ± 0.4, 21.8 ± 0.8 and 28.9 ± 0.8 μg·mL−1, respectively. Antifouling bioassay tests showed that compounds 1, 11, 12, 15 had potent antifouling activity with EC50 values of 12.9 ± 0.5, 22.1 ± 0.8, 7.4 ± 0.6, 16.1 ± 0.6 μg·mL−1 toward barnacle Balanus amphitrite larvae, respectively. PMID:25522319

  12. 40 CFR 721.10172 - Alkylamide derivative (generic).

    Code of Federal Regulations, 2011 CFR

    2011-07-01

    ... 40 Protection of Environment 31 2011-07-01 2011-07-01 false Alkylamide derivative (generic). 721... Substances § 721.10172 Alkylamide derivative (generic). (a) Chemical substance and significant new uses subject to reporting. (1) The chemical substance identified generically as alkylamide derivative (PMN P-03...

  13. 40 CFR 721.10172 - Alkylamide derivative (generic).

    Code of Federal Regulations, 2010 CFR

    2010-07-01

    ... 40 Protection of Environment 30 2010-07-01 2010-07-01 false Alkylamide derivative (generic). 721... Substances § 721.10172 Alkylamide derivative (generic). (a) Chemical substance and significant new uses subject to reporting. (1) The chemical substance identified generically as alkylamide derivative (PMN P-03...

  14. Anisotropic inflation with derivative couplings

    NASA Astrophysics Data System (ADS)

    Holland, Jonathan; Kanno, Sugumi; Zavala, Ivonne

    2018-05-01

    We study anisotropic power-law inflationary solutions when the inflaton and its derivative couple to a vector field. This type of coupling is motivated by D-brane inflationary models, in which the inflaton, and a vector field living on the D-brane, couple disformally (derivatively). We start by studying a phenomenological model where we show the existence of anisotropic solutions and demonstrate their stability via a dynamical system analysis. Compared to the case without a derivative coupling, the anisotropy is reduced and thus can be made consistent with current limits, while the value of the slow-roll parameter remains almost unchanged. We also discuss solutions for more general cases, including D-brane-like couplings.

  15. Transformational derivation of programs using the Focus system

    NASA Technical Reports Server (NTRS)

    Reddy, Uday S.

    1988-01-01

    A program derivation support system called Focus is being constructed. It will formally derive programs using the paradigm of program transformation. The following issues are discussed: (1) the integration of validation and program derivation activities in the Focus system; (2) its tree-based user interface; (3) the control of search spaces in program derivation; and (4) the structure and organization of program derivation records. The inference procedures of the system are based on the integration of functional and logic programming principles. This brings about a synthesis of paradigms that were heretofore considered far apart, such as logical and executable specifications and constructive and transformational approaches to program derivation. A great emphasis has been placed, in the design of Focus, on achieving small search spaces during program derivation. The program manipulation operations such as expansion, simplification and rewriting were designed with this objective. The role of operations that are expensive in search spaces, such as folding, has been reduced. Program derivations are documented in Focus in a way that the high level descriptions of derivations are expressed only using program level information. All the meta-level information, together with dependencies between derivations of program components, is automatically recorded by the system at a lower level of description for its own use in replay.

  16. New steroid derivative with hypoglycemic activity

    PubMed Central

    Lauro, Figueroa-Valverde; Francisco, Díaz-Cedillo; Lenin, Hau-Heredia; Elodia, García-Cervera; Eduardo, Pool-Gómez; Marcela, Rosas-Nexticapa; Bety, Sarabia-Alcocer

    2014-01-01

    Data indicates that some steroid derivatives may induce changes on glucose levels; nevertheless, data are very confusing. Therefore, more pharmacological data are needed to characterize the activity induced by the steroid derivatives on glucose levels. The aim of this study was to synthesize a new steroid derivative for evaluate its hypoglycemic activity. The effects of steroid derivative on glucose concentration were evaluated in a diabetic animal model using glibenclamide and metformin as controls. In addition, the pregnenolone-dihydrotestosterone conjugate was bound to Tc-99m using radioimmunoassay methods, to evaluate the pharmacokinetics of the steroid derivative over time. The results showed that the pregnenolone-dihydrotestosterone conjugate induces changes on the glucose levels in similar form than glibenclamide. Other data showed that the biodistribution of Tc-99m-steroid derivativein brain was higher in comparison with spleen, stomach, intestine liver and kidney. In conclusion, the pregnenolone-dihydrotestosterone conjugate exerts hypoglycemic activity and this phenomenon could depend of its physicochemical properties which could be related to the degree of lipophilicity of the steroidderivative. PMID:25550906

  17. [Separation of [Rh-103m]-rhodocene derivatives from the parent [103Ru]ruthenocene derivatives and their organ distribution].

    PubMed

    Wenzel, M; Wu, Y F

    1987-01-01

    The radioactive decay of [103Ru]ruthenocene derivatives leads to 103mRh labelled rhodocinium derivatives, which can be separated by the extraction of a lipophilic solution of the ruthenocen derivate with water. The separation factor 103mRh/103Ru reaches values of 32:1 Rh3+ ions are not liberated and extracted. The organ distribution of the 103mRh labelled rhodocinium derivatives gained from ruthenocene and from N-isopropyl-ruthenocene amphetamine is different from the distribution of the parent ruthenocene compound. The liver and kidney uptake of the rhodocinium-amphetamine is much higher than the uptake with ruthenocene amphetamine.

  18. Synthesis of ketones from biomass-derived feedstock.

    PubMed

    Meng, Qinglei; Hou, Minqiang; Liu, Huizhen; Song, Jinliang; Han, Buxing

    2017-01-31

    Cyclohexanone and its derivatives are very important chemicals, which are currently produced mainly by oxidation of cyclohexane or alkylcyclohexane, hydrogenation of phenols, and alkylation of cyclohexanone. Here we report that bromide salt-modified Pd/C in H 2 O/CH 2 Cl 2 can efficiently catalyse the transformation of aromatic ethers, which can be derived from biomass, to cyclohexanone and its derivatives via hydrogenation and hydrolysis processes. The yield of cyclohexanone from anisole can reach 96%, and the yields of cyclohexanone derivatives produced from the aromatic ethers, which can be extracted from plants or derived from lignin, are also satisfactory. Detailed study shows that the Pd, bromide salt and H 2 O/CH 2 Cl 2 work cooperatively to promote the desired reaction and inhibit the side reaction. Thus high yields of desired products can be obtained. This work opens the way for production of ketones from aromatic ethers that can be derived from biomass.

  19. Antimicrobial Potential of Benzimidazole Derived Molecules.

    PubMed

    Bansal, Yogita; Kaur, Manjinder; Bansal, Gulshan

    2017-10-31

    Structural resemblance of benzimidazole nucleus with purine nucleus in nucleotides makes benzimidazole derivatives attractive ligands to interact with biopolymers of a living system. The most prominent benzimidazole compound in nature is N-ribosyldimethylbenzimidazole, which serves as an axial ligand for cobalt in vitamin B12. This structural similarity prompted medicinal chemists across the globe to synthesize a variety of benzimidazole derivatives and to screen those for various biological activities, such as anticancer, hormone antagonist, antiviral, anti-HIV, anthelmintic, antiprotozoal, antimicrobial, antihypertensive, anti-inflammatory, analgesic, anxiolytic, antiallergic, coagulant, anticoagulant, antioxidant and antidiabetic activities. Hence, benzimidazole nucleus is considered as a privileged structure in drug discovery, and it is exploited by many research groups to develop numerous compounds that are purported to be antimicrobial. Despite a large volume of research in this area, no novel benzimidazole derived compound has emerged as clinically effective antimicrobial drug. In the present review, we have compiled various reports on benzimidazole derived antimicrobials, classified as monosubstituted, disubstituted, trisubstituted and tetrasubstituted benzimidazoles, bis-benzimidazoles, fused-benzimidazoles, and benzimidazole derivative-metal complexes. The purpose is to collate these research reports, and to generate a generalised outlay of benzimidazole derived molecules that can assist the medicinal chemists in selecting appropriate combination of substituents around the nucleus for designing potent antimicrobials. Copyright© Bentham Science Publishers; For any queries, please email at epub@benthamscience.org.

  20. Derivation of Improved Surface and TOA Broadband Fluxes Using CERES-derived Narrowband-to-Broadband Coefficients

    NASA Technical Reports Server (NTRS)

    Khaiyer, Mandana M.; Doelling, David R.; Chan, Pui K.; Nordeen, MIchele L.; Palikonda, Rabindra; Yi, Yuhong; Minnis, Patrick

    2006-01-01

    Satellites can provide global coverage of a number of climatically important radiative parameters, including broadband (BB) shortwave (SW) and longwave (LW) fluxes at the top of the atmosphere (TOA) and surface. These parameters can be estimated from narrowband (NB) Geostationary Operational Environmental Satellite (GOES) data, but their accuracy is highly dependent on the validity of the narrowband-to-broadband (NB-BB) conversion formulas that are used to convert the NB fluxes to broadband values. The formula coefficients have historically been derived by regressing matched polarorbiting satellite BB fluxes or radiances with their NB counterparts from GOES (e.g., Minnis et al., 1984). More recently, the coefficients have been based on matched Earth Radiation Budget Experiment (ERBE) and GOES-6 data (Minnis and Smith, 1998). The Clouds and the Earth's Radiant Energy Budget (CERES see Wielicki et al. 1998)) project has recently developed much improved Angular Distribution Models (ADM; Loeb et al., 2003) and has higher resolution data compared to ERBE. A limited set of coefficients was also derived from matched GOES-8 and CERES data taken on Topical Rainfall Measuring Mission (TRMM) satellite (Chakrapani et al., 2003; Doelling et al., 2003). The NB-BB coefficients derived from CERES and the GOES suite should yield more accurate BB fluxes than from ERBE, but are limited spatially and seasonally. With CERES data taken from Terra and Aqua, it is now possible to derive more reliable NB-BB coefficients for any given area. Better TOA fluxes should translate to improved surface radiation fluxes derived using various algorithms. As part of an ongoing effort to provide accurate BB flux estimates for the Atmospheric Radiation Measurement (ARM) Program, this paper documents the derivation of new NB-BB coefficients for the ARM Southern Great Plains (SGP) domain and for the Darwin region of the Tropical Western Pacific (DTWP) domain.

  1. ASSESSING UV IRRADIANCE IN CARIBBEAN REEF CORAL AND DNA DAMAGE IN THEIR CORAL AND ZOOXANTHELLAE

    EPA Science Inventory

    UV penetration into the water near coral reefs may be increasing as a consequence of global climate change. Calm waters associated with ENSO conditions can enhance stratification that increases the amount of photobleaching of chromophoric dissolved organic matter (CDOM) in surfa...

  2. Cytotoxic activity of aminoderivatized cationic chitosan derivatives.

    PubMed

    Lee, Jung-Kul; Lim, Hyun-Soo; Kim, Jung-Hoe

    2002-10-21

    Chitosan derivatives were prepared by dialkylaminoalkylation and reductive amination followed by quaternization. In this study, the cytotoxic activity of the chitosan derivatives was investigated and a relationship between structure and activity is suggested. The cationic chitosan derivatives elicited dose-dependent inhibitory effects on the proliferation of tumor cell lines.

  3. 45 CFR 1630.12 - Applicability to derivative income.

    Code of Federal Regulations, 2011 CFR

    2011-10-01

    ... 45 Public Welfare 4 2011-10-01 2011-10-01 false Applicability to derivative income. 1630.12... CORPORATION COST STANDARDS AND PROCEDURES § 1630.12 Applicability to derivative income. (a) Derivative income... activity. (b) Derivative income which is allocated to the LSC fund in accordance with paragraph (a) of this...

  4. 17 CFR 240.16a-4 - Derivative securities.

    Code of Federal Regulations, 2010 CFR

    2010-04-01

    ... 17 Commodity and Securities Exchanges 3 2010-04-01 2010-04-01 false Derivative securities. 240.16a....16a-4 Derivative securities. (a) For purposes of section 16 of the Act, both derivative securities and... securities, except that the acquisition or disposition of any derivative security shall be separately...

  5. A new photostabilizer: Hydrogenated benzoin derivatives

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

    Yamaguchi, K.; Ohkatsu, Y.

    1993-12-31

    It is found that synergistic effects based on combined use of HALS (hindered Amine Light Stabilizers) with phenolic antioxidants consist of the action of HALS as hydrogen donor to quinones, derived from the phenol in autoxidation, excited by uv light. The finding has been realized as a new photostabilizer of hydrogenated benzoin derivatives. They are generally characterized by multifunctions. The o,o`-dihydroxyl-substituted derivatives inter alia extend the life of a phenolic antioxidant co-used, as hydrogen donor, as well as ultimately act as uv absorber.

  6. Multifunctional Cinnamic Acid Derivatives.

    PubMed

    Peperidou, Aikaterini; Pontiki, Eleni; Hadjipavlou-Litina, Dimitra; Voulgari, Efstathia; Avgoustakis, Konstantinos

    2017-07-25

    Our research to discover potential new multitarget agents led to the synthesis of 10 novel derivatives of cinnamic acids and propranolol, atenolol, 1-adamantanol, naphth-1-ol, and (benzylamino) ethan-1-ol. The synthesized molecules were evaluated as trypsin, lipoxygenase and lipid peroxidation inhibitors and for their cytotoxicity. Compound 2b derived from phenoxyphenyl cinnamic acid and propranolol showed the highest lipoxygenase (LOX) inhibition (IC 50 = 6 μΜ) and antiproteolytic activity (IC 50 = 0.425 μΜ). The conjugate 1a of simple cinnamic acid with propranolol showed the higher antiproteolytic activity (IC 50 = 0.315 μΜ) and good LOX inhibitory activity (IC 50 = 66 μΜ). Compounds 3a and 3b , derived from methoxylated caffeic acid present a promising combination of in vitro inhibitory and antioxidative activities. The S isomer of 2b also presented an interesting multitarget biological profile in vitro . Molecular docking studies point to the fact that the theoretical results for LOX-inhibitor binding are identical to those from preliminary in vitro study.

  7. Derivative Sign Patterns in Two Dimensions

    ERIC Educational Resources Information Center

    Schilling, Kenneth

    2013-01-01

    Given a function defined on a subset of the plane whose partial derivatives never change sign, the signs of the partial derivatives form a two-dimensional pattern. We explore what patterns are possible for various planar domains.

  8. 12 CFR 324.34 - OTC derivative contracts.

    Code of Federal Regulations, 2014 CFR

    2014-01-01

    ... calculating PFE. (E) The PFE of the protection provider of a credit derivative is capped at the net present... single OTC derivative contract is the greater of the mark-to-fair value of the OTC derivative contract or... with a negative mark-to-fair value, is calculated by multiplying the notional principal amount of the...

  9. Improved Second Derivative Test for Relative Extrema

    ERIC Educational Resources Information Center

    Wu, Yan

    2007-01-01

    In this note, a modified Second Derivative Test is introduced for the relative extrema of a single variable function. This improved test overcomes the difficulty of the second derivative vanishing at the critical point, while in contrast the traditional test fails for this case. A proof for this improved Second Derivative Test is presented,…

  10. Methods for deoxygenating biomass-derived pyrolysis oil

    DOEpatents

    Baird, Lance Awender; Brandvold, Timothy A.

    2015-06-30

    Methods for deoxygenating a biomass-derived pyrolysis oil are provided. A method for deoxygenating a biomass-derived pyrolysis oil comprising the steps of combining a biomass-derived pyrolysis oil stream with a heated low-oxygen-pyoil diluent recycle stream to form a heated diluted pyoil feed stream is provided. The heated diluted pyoil feed stream has a feed temperature of about 150.degree. C. or greater. The heated diluted pyoil feed stream is contacted with a first deoxygenating catalyst in the presence of hydrogen at first hydroprocessing conditions effective to form a low-oxygen biomass-derived pyrolysis oil effluent.

  11. Magnetic cellulose-derivative structures

    DOEpatents

    Walsh, Myles A.; Morris, Robert S.

    1986-09-16

    Structures to serve as selective magnetic sorbents are formed by dissolving a cellulose derivative such as cellulose triacetate in a solvent containing magnetic particles. The resulting solution is sprayed as a fine mist into a chamber containing a liquid coagulant such as n-hexane in which the cellulose derivative is insoluble but in which the coagulant is soluble or miscible. On contact with the coagulant, the mist forms free-flowing porous magnetic microspheric structures. These structures act as containers for the ion-selective or organic-selective sorption agent of choice. Some sorbtion agents can be incorporated during the manufacture of the structure.

  12. 28 CFR 17.26 - Derivative classification.

    Code of Federal Regulations, 2010 CFR

    2010-07-01

    ... 28 Judicial Administration 1 2010-07-01 2010-07-01 false Derivative classification. 17.26 Section 17.26 Judicial Administration DEPARTMENT OF JUSTICE CLASSIFIED NATIONAL SECURITY INFORMATION AND ACCESS TO CLASSIFIED INFORMATION Classified Information § 17.26 Derivative classification. (a) Persons...

  13. mTOR Is Essential for Corticosteroid Effects on Hippocampal AMPA Receptor Function and Fear Memory

    ERIC Educational Resources Information Center

    Xiong, Hui; Casse, Frédéric; Zhou, Yang; Zhou, Ming; Xiong, Zhi-Qi; Joëls, Marian; Martin, Stéphane; Krugers, Harm J.

    2015-01-01

    Glucocorticoid hormones, via activation of their receptors, promote memory consolidation, but the exact underlying mechanisms remain elusive. We examined how corticosterone regulates AMPA receptors (AMPARs), which are crucial for synaptic plasticity and memory formation. Combining a live imaging fluorescent recovery after photobleaching approach…

  14. Satellite-Derived Bathymetry: Accuracy Assessment on Depths Derivation Algorithm for Shallow Water Area

    NASA Astrophysics Data System (ADS)

    Said, N. M.; Mahmud, M. R.; Hasan, R. C.

    2017-10-01

    Over the years, the acquisition technique of bathymetric data has evolved from a shipborne platform to airborne and presently, utilising space-borne acquisition. The extensive development of remote sensing technology has brought in the new revolution to the hydrographic surveying. Satellite-Derived Bathymetry (SDB), a space-borne acquisition technique which derives bathymetric data from high-resolution multispectral satellite imagery for various purposes recently considered as a new promising technology in the hydrographic surveying industry. Inspiring by this latest developments, a comprehensive study was initiated by National Hydrographic Centre (NHC) and Universiti Teknologi Malaysia (UTM) to analyse SDB as a means for shallow water area acquisition. By adopting additional adjustment in calibration stage, a marginal improvement discovered on the outcomes from both Stumpf and Lyzenga algorithms where the RMSE values for the derived (predicted) depths were 1.432 meters and 1.728 meters respectively. This paper would deliberate in detail the findings from the study especially on the accuracy level and practicality of SDB over the tropical environmental setting in Malaysia.

  15. Contribution of low-temperature single-molecule techniques to structural issues of pigment–protein complexes from photosynthetic purple bacteria

    PubMed Central

    Löhner, Alexander; Cogdell, Richard

    2018-01-01

    As the electronic energies of the chromophores in a pigment–protein complex are imposed by the geometrical structure of the protein, this allows the spectral information obtained to be compared with predictions derived from structural models. Thereby, the single-molecule approach is particularly suited for the elucidation of specific, distinctive spectral features that are key for a particular model structure, and that would not be observable in ensemble-averaged spectra due to the heterogeneity of the biological objects. In this concise review, we illustrate with the example of the light-harvesting complexes from photosynthetic purple bacteria how results from low-temperature single-molecule spectroscopy can be used to discriminate between different structural models. Thereby the low-temperature approach provides two advantages: (i) owing to the negligible photobleaching, very long observation times become possible, and more importantly, (ii) at cryogenic temperatures, vibrational degrees of freedom are frozen out, leading to sharper spectral features and in turn to better resolved spectra. PMID:29321265

  16. Highly Stable Near-Infrared Fluorescent Organic Nanoparticles with a Large Stokes Shift for Noninvasive Long-Term Cellular Imaging.

    PubMed

    Zhang, Jinfeng; Chen, Rui; Zhu, Zelin; Adachi, Chihaya; Zhang, Xiaohong; Lee, Chun-Sing

    2015-12-02

    Fluorescent organic nanoparticles based on small molecules have been regarded as promising candidates for bioimaging in recent years. In this study, we report a highly stable near-infrared (NIR) fluorescent organic nanoprobes based on nanoparticles of an anthraquinone derivate with strong aggregation-induced emission (AIE) characteristics and a large Stokes shift (>175 nm). These endow the nanoprobe with high fluorescent brightness and high signal-to-noise ratio. On the other hand, the nanoprobe also shows low cytotoxicity, good stability over a wide pH range, superior resistance against photodegradation and photobleaching comparing to typical commercial fluorescent organic dyes such as fluorescein sodium. Endowed with such merits in term of optical performance, biocompatibility, and stability, the nanoprobe is demonstrated to be an ideal fluorescent probe for noninvasive long-term cellular tracing and imaging applications. As an example, it is shown that strong red fluorescence from the nanoprobe can still be clearly observed in A549 human lung cancer cells after incubation for six generations over 15 days.

  17. Determining absolute protein numbers by quantitative fluorescence microscopy.

    PubMed

    Verdaasdonk, Jolien Suzanne; Lawrimore, Josh; Bloom, Kerry

    2014-01-01

    Biological questions are increasingly being addressed using a wide range of quantitative analytical tools to examine protein complex composition. Knowledge of the absolute number of proteins present provides insights into organization, function, and maintenance and is used in mathematical modeling of complex cellular dynamics. In this chapter, we outline and describe three microscopy-based methods for determining absolute protein numbers--fluorescence correlation spectroscopy, stepwise photobleaching, and ratiometric comparison of fluorescence intensity to known standards. In addition, we discuss the various fluorescently labeled proteins that have been used as standards for both stepwise photobleaching and ratiometric comparison analysis. A detailed procedure for determining absolute protein number by ratiometric comparison is outlined in the second half of this chapter. Counting proteins by quantitative microscopy is a relatively simple yet very powerful analytical tool that will increase our understanding of protein complex composition. © 2014 Elsevier Inc. All rights reserved.

  18. Generation-3 programmable array microscope (PAM) with digital micro-mirror device (DMD)

    NASA Astrophysics Data System (ADS)

    De Beule, Pieter A. A.; de Vries, Anthony H. B.; Arndt-Jovin, Donna J.; Jovin, Thomas M.

    2011-03-01

    We report progress on the construction of an optical sectioning programmable array microscope (PAM) implemented with a digital micro-mirror device (DMD) spatial light modulator (SLM) utilized for both fluorescence illumination and detection. The introduction of binary intensity modulation at the focal plane of a microscope objective in a computer controlled pixilated mode allows the recovery of an optically sectioned image. Illumination patterns can be changed very quickly, in contrast to static Nipkow disk or aperture correlation implementations, thereby creating an optical system that can be optimized to the optical specimen in a convenient manner, e.g. for patterned photobleaching, photobleaching reduction, or spatial superresolution. We present a third generation (Gen-3) dual path PAM module incorporating the 25 kHz binary frame rate TI 1080p DMD and a newly developed optical system that offers diffraction limited imaging with compensation of tilt angle distortion.

  19. Interface Defect States and Charge Transport Properties in Low-Cost Photovoltaic Devices made from Scalable Deposition Methods

    NASA Astrophysics Data System (ADS)

    Marin, Andrew; Munoz-Rojas, David; Iza, Diana; Gershon, Talia; MacManus-Driscoll, Judith

    2011-03-01

    In-plane (parallel to the substrate) polymer diffusion at and near interfaces has significant implications for polymeric surfactants used in tertiary oil recovery, exfoliation of clay sheets in polymer nano-composites, and several other high technology applications. Here, we report a study on the in-plane diffusion of whole polymer chains confined between interfaces using fluorescence recovery after photobleaching. Adapted from quantitative biology, FRAP provides a platform to independently study the effect of temperature, molecular weight, and film thickness on in-plane diffusion of polymers confined between interfaces. Fluorescently labeled polymers were synthesized, spin coated onto quartz substrates and the self-diffusion coefficient was measured by irreversibly photobleaching fluorophores in a pre-defined pattern and monitoring recovery of fluorescence over time. Preliminary results indicate that for thick films the diffusion coefficient is consistent with bulk values. The authors would like to thank the Gates-Cambridge Trust and the International Copper Association.

  20. 17 CFR 200.506 - Derivative classification.

    Code of Federal Regulations, 2012 CFR

    2012-04-01

    ... 17 Commodity and Securities Exchanges 2 2012-04-01 2012-04-01 false Derivative classification. 200...; CONDUCT AND ETHICS; AND INFORMATION AND REQUESTS Classification and Declassification of National Security Information and Material § 200.506 Derivative classification. Any document that includes paraphrases...

  1. 22 CFR 9.6 - Derivative classification.

    Code of Federal Regulations, 2012 CFR

    2012-04-01

    ... 22 Foreign Relations 1 2012-04-01 2012-04-01 false Derivative classification. 9.6 Section 9.6... classification. (a) Definition. Derivative classification is the incorporating, paraphrasing, restating or... with the classification of the source material. Duplication or reproduction of existing classified...

  2. 17 CFR 200.506 - Derivative classification.

    Code of Federal Regulations, 2013 CFR

    2013-04-01

    ... 17 Commodity and Securities Exchanges 2 2013-04-01 2013-04-01 false Derivative classification. 200...; CONDUCT AND ETHICS; AND INFORMATION AND REQUESTS Classification and Declassification of National Security Information and Material § 200.506 Derivative classification. Any document that includes paraphrases...

  3. 22 CFR 9.6 - Derivative classification.

    Code of Federal Regulations, 2013 CFR

    2013-04-01

    ... 22 Foreign Relations 1 2013-04-01 2013-04-01 false Derivative classification. 9.6 Section 9.6... classification. (a) Definition. Derivative classification is the incorporating, paraphrasing, restating or... with the classification of the source material. Duplication or reproduction of existing classified...

  4. 17 CFR 200.506 - Derivative classification.

    Code of Federal Regulations, 2014 CFR

    2014-04-01

    ... 17 Commodity and Securities Exchanges 3 2014-04-01 2014-04-01 false Derivative classification. 200...; CONDUCT AND ETHICS; AND INFORMATION AND REQUESTS Classification and Declassification of National Security Information and Material § 200.506 Derivative classification. Any document that includes paraphrases...

  5. 22 CFR 9.6 - Derivative classification.

    Code of Federal Regulations, 2014 CFR

    2014-04-01

    ... 22 Foreign Relations 1 2014-04-01 2014-04-01 false Derivative classification. 9.6 Section 9.6... classification. (a) Definition. Derivative classification is the incorporating, paraphrasing, restating or... with the classification of the source material. Duplication or reproduction of existing classified...

  6. 22 CFR 9.6 - Derivative classification.

    Code of Federal Regulations, 2011 CFR

    2011-04-01

    ... 22 Foreign Relations 1 2011-04-01 2011-04-01 false Derivative classification. 9.6 Section 9.6... classification. (a) Definition. Derivative classification is the incorporating, paraphrasing, restating or... with the classification of the source material. Duplication or reproduction of existing classified...

  7. 17 CFR 200.506 - Derivative classification.

    Code of Federal Regulations, 2011 CFR

    2011-04-01

    ... 17 Commodity and Securities Exchanges 2 2011-04-01 2011-04-01 false Derivative classification. 200...; CONDUCT AND ETHICS; AND INFORMATION AND REQUESTS Classification and Declassification of National Security Information and Material § 200.506 Derivative classification. Any document that includes paraphrases...

  8. Catalytic combustion of coal-derived liquids

    NASA Technical Reports Server (NTRS)

    Bulzan, D. L.; Tacina, R. R.

    1981-01-01

    A noble metal catalytic reactor was tested with three grades of SRC 2 coal derived liquids, naphtha, middle distillate, and a blend of three parts middle distillate to one part heavy distillate. A petroleum derived number 2 diesel fuel was also tested to provide a direct comparison. The catalytic reactor was tested at inlet temperatures from 600 to 800 K, reference velocities from 10 to 20 m/s, lean fuel air ratios, and a pressure of 3 x 10 to the 5th power Pa. Compared to the diesel, the naphtha gave slightly better combustion efficiency, the middle distillate was almost identical, and the middle heavy blend was slightly poorer. The coal derived liquid fuels contained from 0.58 to 0.95 percent nitrogen by weight. Conversion of fuel nitrogen to NOx was approximately 75 percent for all three grades of the coal derived liquids.

  9. Daryamide Analogues from a Marine-Derived Streptomyces species.

    PubMed

    Fu, Peng; La, Scott; MacMillan, John B

    2017-04-28

    Three new cyclohexene amine derivatives, daryamides D-F (1-3), a new arylamine derivative, carpatamide D (4), and a new ornithine lactamization derivative, ornilactam A (5), were isolated from the marine-derived Streptomyces strain SNE-011. Their structures, including absolute configurations, were elucidated on the basis of spectroscopic analysis and chemical methods. The carpatamide skeleton could be considered as the biosynthetic precursor of the daryamides.

  10. 5 CFR 2500.5 - Derivative classification.

    Code of Federal Regulations, 2014 CFR

    2014-01-01

    ... 5 Administrative Personnel 3 2014-01-01 2014-01-01 false Derivative classification. 2500.5 Section 2500.5 Administrative Personnel OFFICE OF ADMINISTRATION, EXECUTIVE OFFICE OF THE PRESIDENT INFORMATION SECURITY REGULATION § 2500.5 Derivative classification. The Office of Administration serves only as the...

  11. 5 CFR 2500.5 - Derivative classification.

    Code of Federal Regulations, 2013 CFR

    2013-01-01

    ... 5 Administrative Personnel 3 2013-01-01 2013-01-01 false Derivative classification. 2500.5 Section 2500.5 Administrative Personnel OFFICE OF ADMINISTRATION, EXECUTIVE OFFICE OF THE PRESIDENT INFORMATION SECURITY REGULATION § 2500.5 Derivative classification. The Office of Administration serves only as the...

  12. 5 CFR 2500.5 - Derivative classification.

    Code of Federal Regulations, 2012 CFR

    2012-01-01

    ... 5 Administrative Personnel 3 2012-01-01 2012-01-01 false Derivative classification. 2500.5 Section 2500.5 Administrative Personnel OFFICE OF ADMINISTRATION, EXECUTIVE OFFICE OF THE PRESIDENT INFORMATION SECURITY REGULATION § 2500.5 Derivative classification. The Office of Administration serves only as the...

  13. 5 CFR 2500.5 - Derivative classification.

    Code of Federal Regulations, 2011 CFR

    2011-01-01

    ... 5 Administrative Personnel 3 2011-01-01 2011-01-01 false Derivative classification. 2500.5 Section 2500.5 Administrative Personnel OFFICE OF ADMINISTRATION, EXECUTIVE OFFICE OF THE PRESIDENT INFORMATION SECURITY REGULATION § 2500.5 Derivative classification. The Office of Administration serves only as the...

  14. PROCEDURES FOR THE DERIVATION OF EQUILIBRIUM ...

    EPA Pesticide Factsheets

    This equilibrium partitioning sediment benchmark (ESB) document describes procedures to derive concentrations for 32 nonionic organic chemicals in sediment which are protective of the presence of freshwater and marine benthic organisms. The equilibrium partitioning (EqP) approach was chosen because it accounts for the varying biological availability of chemicals in different sediments and allows for the incorporation of the appropriate biological effects concentration. This provides for the derivation of benchmarks that are causally linked to the specific chemical, applicable across sediments, and appropriately protective of benthic organisms. EqP can be used to calculate ESBs for any toxicity endpoint for which there are water-only toxicity data; it is not limited to any single effect endpoint. For the purposes of this document, ESBs for 32 nonionic organic chemicals, including several low molecular weight aliphatic and aromatic compounds, pesticides, and phthalates, were derived using Final Chronic Values (FCV) from Water Quality Criteria (WQC) or Secondary Chronic Values (SCV) derived from existing toxicological data using the Great Lakes Water Quality Initiative (GLI) or narcosis theory approaches. These values are intended to be the concentration of each chemical in water that is protective of the presence of aquatic life. For nonionic organic chemicals demonstrating a narcotic mode of action, ESBs derived using the GLI approach specifically for fres

  15. Exchange of rotor components in functioning bacterial flagellar motor

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

    Fukuoka, Hajime; Inoue, Yuichi; Graduate School of Life Sciences, Tohoku University, Aoba-ku, Sendai 980-8577

    2010-03-26

    The bacterial flagellar motor is a rotary motor driven by the electrochemical potential of a coupling ion. The interaction between a rotor and stator units is thought to generate torque. The overall structure of flagellar motor has been thought to be static, however, it was recently proved that stators are exchanged in a rotating motor. Understanding the dynamics of rotor components in functioning motor is important for the clarifying of working mechanism of bacterial flagellar motor. In this study, we focused on the dynamics and the turnover of rotor components in a functioning flagellar motor. Expression systems for GFP-FliN, FliM-GFP,more » and GFP-FliG were constructed, and each GFP-fusion was functionally incorporated into the flagellar motor. To investigate whether the rotor components are exchanged in a rotating motor, we performed fluorescence recovery after photobleaching experiments using total internal reflection fluorescence microscopy. After photobleaching, in a tethered cell producing GFP-FliN or FliM-GFP, the recovery of fluorescence at the rotational center was observed. However, in a cell producing GFP-FliG, no recovery of fluorescence was observed. The transition phase of fluorescence intensity after full or partially photobleaching allowed the turnover of FliN subunits to be calculated as 0.0007 s{sup -1}, meaning that FliN would be exchanged in tens of minutes. These novel findings indicate that a bacterial flagellar motor is not a static structure even in functioning state. This is the first report for the exchange of rotor components in a functioning bacterial flagellar motor.« less

  16. Methods for deoxygenating biomass-derived pyrolysis oil

    DOEpatents

    Brandvold, Timothy A.

    2015-07-14

    Methods for deoxygenating a biomass-derived pyrolysis oil are provided. A method comprising the steps of diluting the biomass-derived pyrolysis oil with a phenolic-containing diluent to form a diluted pyoil-phenolic feed is provided. The diluted pyoil-phenolic feed is contacted with a deoxygenating catalyst in the presence of hydrogen at hydroprocessing conditions effective to form a low-oxygen biomass-derived pyrolysis oil effluent.

  17. Transportation Planning with Immune System Derived Approach

    NASA Astrophysics Data System (ADS)

    Sugiyama, Kenji; Yaji, Yasuhito; Ootsuki, John Takuya; Fujimoto, Yasutaka; Sekiguchi, Takashi

    This paper presents an immune system derived approach for planning transportation of materials between manufacturing processes in the factory. Transportation operations are modeled by Petri Net, and divided into submodels. Transportation orders are derived from the firing sequences of those submodels through convergence calculation by the immune system derived excitation and suppression operations. Basic evaluation of this approach is conducted by simulation-based investigation.

  18. Reactions of glycidyl derivatives with ambident nucleophiles; part 2: amino acid derivatives

    PubMed Central

    Dyker, Gerald; Thöne, Andreas; Henkel, Gerald

    2007-01-01

    A three-step procedure for the synthesis of multifunctionalized heterocycles from a pyroglutamic acid derivative, glycidyl components and anilines by nucleophilic substitution and cobalt catalysis is presented. PMID:17900352

  19. Calculation of Sensitivity Derivatives in an MDAO Framework

    NASA Technical Reports Server (NTRS)

    Moore, Kenneth T.

    2012-01-01

    During gradient-based optimization of a system, it is necessary to generate the derivatives of each objective and constraint with respect to each design parameter. If the system is multidisciplinary, it may consist of a set of smaller "components" with some arbitrary data interconnection and process work ow. Analytical derivatives in these components can be used to improve the speed and accuracy of the derivative calculation over a purely numerical calculation; however, a multidisciplinary system may include both components for which derivatives are available and components for which they are not. Three methods to calculate the sensitivity of a mixed multidisciplinary system are presented: the finite difference method, where the derivatives are calculated numerically; the chain rule method, where the derivatives are successively cascaded along the system's network graph; and the analytic method, where the derivatives come from the solution of a linear system of equations. Some improvements to these methods, to accommodate mixed multidisciplinary systems, are also presented; in particular, a new method is introduced to allow existing derivatives to be used inside of finite difference. All three methods are implemented and demonstrated in the open-source MDAO framework OpenMDAO. It was found that there are advantages to each of them depending on the system being solved.

  20. Resveratrol derivatives as a pharmacological tool.

    PubMed

    Biasutto, Lucia; Mattarei, Andrea; Azzolini, Michele; La Spina, Martina; Sassi, Nicola; Romio, Matteo; Paradisi, Cristina; Zoratti, Mario

    2017-09-01

    Prodrugs of resveratrol are under development. Among the long-term goals, still largely elusive, are (1) modulating physical properties (e.g., water-soluble derivatives bearing polyethylene glycol chains), (2) changing distribution in the body (e.g., galactosyl derivatives restricted to the intestinal lumen), (3) increasing absorption from the gastrointestinal tract (e.g., derivatives imitating the natural substrates of endogenous transporters), and (4) hindering phase II metabolism (e.g., temporarily blocking the hydroxyls), all contributing to (5) increasing bioavailability. The chemical bonds that have been tested for functionalization include carboxyester, acetal, and carbamate groups. A second approach, which can be combined with the first, seeks to reinforce or modify the biochemical activities of resveratrol by concentrating the compound at specific subcellular sites. An example is provided by mitochondria-targeted derivatives. These proved to be pro-oxidant and cytotoxic in vitro, selectively killing fast-growing and tumor cells when supplied in the low micromolar range. This suggests the possibility of anticancer applications. © 2017 New York Academy of Sciences.

  1. Coal-derived humus: plant growth effects

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

    Freeman, P.G.; Fowkes, W.W.

    The growth-promoting properties of coal-derived humus were tested by a variety of bioassay methods. Humate materials used included leonardite, a form of naturally oxidized lignite, and humic acids extracted from leonardite. It was found that the greatest effects were evident when water-soluble humic acid was applied to excised root tips. No significant short-term effect was noted when water-soluble humic acid was applied to stem or coleoptile sections. Root initiation was promoted in bean stems when high concentrations of coal-derived humate were applied. Gross weight of tomato plants was increased by up to 40 percent when coal-derived humates were added inmore » concentrations of 10/sup 1/ to 10/sup 4/ ppM to the nutrient in a hydroponic growth test of 6 weeks' duration. It is concluded that coal-derived humate promotes the uptake of minerals, specifically iron, by serving as a metal chelate.« less

  2. Nanomaterials derived from metal-organic frameworks

    NASA Astrophysics Data System (ADS)

    Dang, Song; Zhu, Qi-Long; Xu, Qiang

    2018-01-01

    The thermal transformation of metal-organic frameworks (MOFs) generates a variety of nanostructured materials, including carbon-based materials, metal oxides, metal chalcogenides, metal phosphides and metal carbides. These derivatives of MOFs have characteristics such as high surface areas, permanent porosities and controllable functionalities that enable their good performance in sensing, gas storage, catalysis and energy-related applications. Although progress has been made to tune the morphologies of MOF-derived structures at the nanometre scale, it remains crucial to further our knowledge of the relationship between morphology and performance. In this Review, we summarize the synthetic strategies and optimized methods that enable control over the size, morphology, composition and structure of the derived nanomaterials. In addition, we compare the performance of materials prepared by the MOF-templated strategy and other synthetic methods. Our aim is to reveal the relationship between the morphology and the physico-chemical properties of MOF-derived nanostructures to optimize their performance for applications such as sensing, catalysis, and energy storage and conversion.

  3. Magnetic cellulose-derivative structures

    DOEpatents

    Walsh, M.A.; Morris, R.S.

    1986-09-16

    Structures to serve as selective magnetic sorbents are formed by dissolving a cellulose derivative such as cellulose triacetate in a solvent containing magnetic particles. The resulting solution is sprayed as a fine mist into a chamber containing a liquid coagulant such as n-hexane in which the cellulose derivative is insoluble but in which the coagulant is soluble or miscible. On contact with the coagulant, the mist forms free-flowing porous magnetic microspheric structures. These structures act as containers for the ion-selective or organic-selective sorption agent of choice. Some sorption agents can be incorporated during the manufacture of the structure. 3 figs.

  4. Toward a better understanding of helicopter stability derivatives

    NASA Technical Reports Server (NTRS)

    Hansen, R. S.

    1982-01-01

    An amended six degree of freedom helicopter stability and control derivative model was developed in which body acceleration and control rate derivatives were included in the Taylor series expansion. These additional derivatives were derived from consideration of the effects of the higher order rotor flapping dynamics, which are known to be inadequately represented in the conventional six degree of freedom, quasistatic stability derivative model. The amended model was a substantial improvement over the conventional model, effectively doubling the unsable bandwidth and providing a more accurate representation of the short period and cross axis characteristics. Further investigations assessed the applicability of the two stability derivative model structures for flight test parameter identification. Parameters were identified using simulation data generated from a higher order base line model having sixth order rotor tip path plane dynamics. Three lower order models were identified: one using the conventional stability derivative model structure, a second using the amended six degree of freedom model structure, and a third model having eight degrees of freedom that included a simplified rotor tip path plane tilt representation.

  5. Ghost-Free Theory with Third-Order Time Derivatives

    NASA Astrophysics Data System (ADS)

    Motohashi, Hayato; Suyama, Teruaki; Yamaguchi, Masahide

    2018-06-01

    As the first step to extend our understanding of higher-derivative theories, within the framework of analytic mechanics of point particles, we construct a ghost-free theory involving third-order time derivatives in Lagrangian. While eliminating linear momentum terms in the Hamiltonian is necessary and sufficient to kill the ghosts associated with higher derivatives for Lagrangian with at most second-order derivatives, we find that this is necessary but not sufficient for the Lagrangian with higher than second-order derivatives. We clarify a set of ghost-free conditions under which we show that the Hamiltonian is bounded, and that equations of motion are reducible into a second-order system.

  6. Benzene derivatives produced by Fusarium graminearum - Short communication.

    PubMed

    Ntushelo, Khayalethu; Setshedi, Itumeleng

    2015-06-01

    Using NMR spectroscopy benzene derivatives were detected in mycelia of Fusarium graminearum, a pathogen of wheat and maize. In previous studies F. graminearum was found to cause cancer to humans and benzene derivatives were detected in breath of cancer sufferers. Surprisingly, no study found benzene derivatives to be the cancerous agents in F. graminearum. In this study we detected benzene derivatives in F. graminearum and propose to study their role as cancer agents.

  7. 18 CFR 367.2440 - Account 244, Derivative instrument liabilities.

    Code of Federal Regulations, 2011 CFR

    2011-04-01

    ... 18 Conservation of Power and Water Resources 1 2011-04-01 2011-04-01 false Account 244, Derivative..., Derivative instrument liabilities. This account must include the change in the fair value of all derivative... the fair value of the derivative instrument. ...

  8. 40 CFR 721.2025 - Substituted phenylimino carbamate derivative.

    Code of Federal Regulations, 2010 CFR

    2010-07-01

    ... derivative. 721.2025 Section 721.2025 Protection of Environment ENVIRONMENTAL PROTECTION AGENCY (CONTINUED... Specific Chemical Substances § 721.2025 Substituted phenylimino carbamate derivative. (a) Chemical... as a substituted phenylimino carbamate derivative (PMN P-91-487) is subject to reporting under this...

  9. 78 FR 66639 - Modifications of Certain Derivative Contracts

    Federal Register 2010, 2011, 2012, 2013, 2014

    2013-11-06

    ... Modifications of Certain Derivative Contracts AGENCY: Internal Revenue Service (IRS), Treasury. ACTION: Final... to the transfer or assignment of certain derivative contracts. The final regulations provide guidance to the nonassigning counterparty to a derivative contract and an assignee on certain notional...

  10. 40 CFR 721.2025 - Substituted phenylimino carbamate derivative.

    Code of Federal Regulations, 2011 CFR

    2011-07-01

    ... derivative. 721.2025 Section 721.2025 Protection of Environment ENVIRONMENTAL PROTECTION AGENCY (CONTINUED... Specific Chemical Substances § 721.2025 Substituted phenylimino carbamate derivative. (a) Chemical... as a substituted phenylimino carbamate derivative (PMN P-91-487) is subject to reporting under this...

  11. 46 CFR 503.55 - Derivative classification.

    Code of Federal Regulations, 2011 CFR

    2011-10-01

    ... 46 Shipping 9 2011-10-01 2011-10-01 false Derivative classification. 503.55 Section 503.55... Security Program § 503.55 Derivative classification. (a) In accordance with Part 2 of Executive Order 13526... developed material consistent with the classification markings that apply to the source information, is...

  12. 46 CFR 503.55 - Derivative classification.

    Code of Federal Regulations, 2010 CFR

    2010-10-01

    ... 46 Shipping 9 2010-10-01 2010-10-01 false Derivative classification. 503.55 Section 503.55... Security Program § 503.55 Derivative classification. (a) In accordance with Part 2 of Executive Order 12958... developed material consistent with the classification markings that apply to the source information, is...

  13. 46 CFR 503.55 - Derivative classification.

    Code of Federal Regulations, 2014 CFR

    2014-10-01

    ... 46 Shipping 9 2014-10-01 2014-10-01 false Derivative classification. 503.55 Section 503.55... Security Program § 503.55 Derivative classification. (a) In accordance with Part 2 of Executive Order 13526... developed material consistent with the classification markings that apply to the source information, is...

  14. 46 CFR 503.55 - Derivative classification.

    Code of Federal Regulations, 2013 CFR

    2013-10-01

    ... 46 Shipping 9 2013-10-01 2013-10-01 false Derivative classification. 503.55 Section 503.55... Security Program § 503.55 Derivative classification. (a) In accordance with Part 2 of Executive Order 13526... developed material consistent with the classification markings that apply to the source information, is...

  15. 46 CFR 503.55 - Derivative classification.

    Code of Federal Regulations, 2012 CFR

    2012-10-01

    ... 46 Shipping 9 2012-10-01 2012-10-01 false Derivative classification. 503.55 Section 503.55... Security Program § 503.55 Derivative classification. (a) In accordance with Part 2 of Executive Order 13526... developed material consistent with the classification markings that apply to the source information, is...

  16. Precision Orbit Derived Atmospheric Density: Development and Performance

    NASA Astrophysics Data System (ADS)

    McLaughlin, C.; Hiatt, A.; Lechtenberg, T.; Fattig, E.; Mehta, P.

    2012-09-01

    Precision orbit ephemerides (POE) are used to estimate atmospheric density along the orbits of CHAMP (Challenging Minisatellite Payload) and GRACE (Gravity Recovery and Climate Experiment). The densities are calibrated against accelerometer derived densities and considering ballistic coefficient estimation results. The 14-hour density solutions are stitched together using a linear weighted blending technique to obtain continuous solutions over the entire mission life of CHAMP and through 2011 for GRACE. POE derived densities outperform the High Accuracy Satellite Drag Model (HASDM), Jacchia 71 model, and NRLMSISE-2000 model densities when comparing cross correlation and RMS with accelerometer derived densities. Drag is the largest error source for estimating and predicting orbits for low Earth orbit satellites. This is one of the major areas that should be addressed to improve overall space surveillance capabilities; in particular, catalog maintenance. Generally, density is the largest error source in satellite drag calculations and current empirical density models such as Jacchia 71 and NRLMSISE-2000 have significant errors. Dynamic calibration of the atmosphere (DCA) has provided measurable improvements to the empirical density models and accelerometer derived densities of extremely high precision are available for a few satellites. However, DCA generally relies on observations of limited accuracy and accelerometer derived densities are extremely limited in terms of measurement coverage at any given time. The goal of this research is to provide an additional data source using satellites that have precision orbits available using Global Positioning System measurements and/or satellite laser ranging. These measurements strike a balance between the global coverage provided by DCA and the precise measurements of accelerometers. The temporal resolution of the POE derived density estimates is around 20-30 minutes, which is significantly worse than that of accelerometer

  17. 17 CFR 200.507 - Declassification dates on derivative documents.

    Code of Federal Regulations, 2011 CFR

    2011-04-01

    ... derivative documents. 200.507 Section 200.507 Commodity and Securities Exchanges SECURITIES AND EXCHANGE... of National Security Information and Material § 200.507 Declassification dates on derivative... derivative document that derives its classification from the approved use of the classification guide of...

  18. Anthraquinones and Derivatives from Marine-Derived Fungi: Structural Diversity and Selected Biological Activities

    PubMed Central

    Fouillaud, Mireille; Venkatachalam, Mekala; Girard-Valenciennes, Emmanuelle; Caro, Yanis; Dufossé, Laurent

    2016-01-01

    Anthraquinones and their derivatives constitute a large group of quinoid compounds with about 700 molecules described. They are widespread in fungi and their chemical diversity and biological activities recently attracted attention of industries in such fields as pharmaceuticals, clothes dyeing, and food colorants. Their positive and/or negative effect(s) due to the 9,10-anthracenedione structure and its substituents are still not clearly understood and their potential roles or effects on human health are today strongly discussed among scientists. As marine microorganisms recently appeared as producers of an astonishing variety of structurally unique secondary metabolites, they may represent a promising resource for identifying new candidates for therapeutic drugs or daily additives. Within this review, we investigate the present knowledge about the anthraquinones and derivatives listed to date from marine-derived filamentous fungi′s productions. This overview highlights the molecules which have been identified in microorganisms for the first time. The structures and colors of the anthraquinoid compounds come along with the known roles of some molecules in the life of the organisms. Some specific biological activities are also described. This may help to open doors towards innovative natural substances. PMID:27023571

  19. Derived Transformation of Children's Pregambling Game Playing

    PubMed Central

    Dymond, Simon; Bateman, Helena; Dixon, Mark R

    2010-01-01

    Contemporary behavior-analytic perspectives on gambling emphasize the impact of verbal relations, or derived relational responding and the transformation of stimulus functions, on the initiation and maintenance of gambling. Approached in this way, it is possible to undertake experimental analysis of the role of verbal/mediational variables in gambling behavior. The present study therefore sought to demonstrate the ways new stimuli could come to have functions relevant to gambling without those functions being trained directly. Following a successful derived-equivalence-relations test, a simulated board game established high- and low-roll functions for two concurrently presented dice labelled with members of the derived relations. During the test for derived transformation, children were reexposed to the board game with dice labelled with indirectly related stimuli. All participants except 1 who passed the equivalence relations test selected the die that was indirectly related to the trained high-roll die more often than the die that was indirectly related to low-roll die, despite the absence of differential outcomes. All participants except 3 also gave the derived high-roll die higher liking ratings than the derived low-roll die. The implications of the findings for behavior-analytic research on gambling and the development of verbally-based interventions for disordered gambling are discussed. PMID:21541176

  20. Derived transformation of children's pregambling game playing.

    PubMed

    Dymond, Simon; Bateman, Helena; Dixon, Mark R

    2010-11-01

    Contemporary behavior-analytic perspectives on gambling emphasize the impact of verbal relations, or derived relational responding and the transformation of stimulus functions, on the initiation and maintenance of gambling. Approached in this way, it is possible to undertake experimental analysis of the role of verbal/mediational variables in gambling behavior. The present study therefore sought to demonstrate the ways new stimuli could come to have functions relevant to gambling without those functions being trained directly. Following a successful derived-equivalence-relations test, a simulated board game established high- and low-roll functions for two concurrently presented dice labelled with members of the derived relations. During the test for derived transformation, children were reexposed to the board game with dice labelled with indirectly related stimuli. All participants except 1 who passed the equivalence relations test selected the die that was indirectly related to the trained high-roll die more often than the die that was indirectly related to low-roll die, despite the absence of differential outcomes. All participants except 3 also gave the derived high-roll die higher liking ratings than the derived low-roll die. The implications of the findings for behavior-analytic research on gambling and the development of verbally-based interventions for disordered gambling are discussed.

  1. Anti-Inflammatory Activity of Tanzawaic Acid Derivatives from a Marine-Derived Fungus Penicillium steckii 108YD142

    PubMed Central

    Shin, Hee Jae; Pil, Gam Bang; Heo, Soo-Jin; Lee, Hyi-Seung; Lee, Jong Seok; Lee, Yeon-Ju; Lee, Jihoon; Won, Ho Shik

    2016-01-01

    Chemical investigation of a marine-derived fungus, Penicillium steckii 108YD142, resulted in the discovery of a new tanzawaic acid derivative, tanzawaic acid Q (1), together with four known analogues, tanzawaic acids A (2), C (3), D (4), and K (5). The structures of tanzawaic acid derivatives 1–5 were determined by the detailed analysis of 1D, 2D NMR and LC-MS data, along with chemical methods and literature data analysis. These compounds significantly inhibited nitric oxide (NO) production and the new tanzawaic acid Q (1) inhibited the lipopolysaccharide (LPS)-induced inducible nitric oxide synthase (iNOS) and cyclooxygenase-2 (COX-2) proteins and mRNA expressions in RAW 264.7 macrophages. Additionally, compound 1 reduced the mRNA levels of inflammatory cytokines. Taken together, the results of this study demonstrated that the new tanzawaic acid derivative inhibits LPS-induced inflammation. This is the first report on the anti-inflammatory activity of tanzawaic acid Q (1). PMID:26761016

  2. Derivative financial instruments and nonprofit health care providers.

    PubMed

    Stewart, Louis J; Owhoso, Vincent

    2004-01-01

    This article examines the extent of derivative financial instrument use among US nonprofit health systems and the impact of these financial instruments on their cash flows, reported operating results, and financial risks. Our examination is conducted through a case study of New Jersey hospitals and health systems. We review the existing literature on interest rate derivative instruments and US hospitals and health systems. This literature describes the design of these derivative financial instruments and the theoretical benefits of their use by large health care provider organizations. Our contribution to the literature is to provide an empirical evaluation of derivative financial instruments usage among a geographically limited sample of US nonprofit health systems. We reviewed the audited financial statements of the 49 community hospitals and multi-hospital health systems operating in the state of New Jersey. We found that 8 percent of New Jersey's nonprofit health providers utilized interest rate derivatives with an aggregate principle value of $229 million. These derivative users combine interest rate swaps and caps to lower the effective interest costs of their long-term debt while limiting their exposure to future interest rate increases. In addition, while derivative assets and liabilities have an immaterial balance sheet impact, derivative related gains and losses are a material component of their reported operating results. We also found that derivative usage among these four health systems was responsible for generating positive cash flows in the range of 1 percent to 2 percent of their total 2001 cash flows from operations. As a result of our admittedly limited samples we conclude that interest rate swaps and caps are effective risk management tools. However, we also found that while these derivative financial instruments are useful hedges against the risks of issuing long-term financing instruments, they also expose derivative users to credit, contract

  3. 40 CFR 721.646 - Aminofluoran derivative (generic name).

    Code of Federal Regulations, 2011 CFR

    2011-07-01

    ... 40 Protection of Environment 31 2011-07-01 2011-07-01 false Aminofluoran derivative (generic name... Substances § 721.646 Aminofluoran derivative (generic name). (a) Chemical substance and significant new uses subject to reporting. (1) The chemical substance identified generically as aminofluoran derivative (PMN P...

  4. 40 CFR 721.9079 - Dihydro quinacridone derivative (generic).

    Code of Federal Regulations, 2011 CFR

    2011-07-01

    ... 40 Protection of Environment 31 2011-07-01 2011-07-01 false Dihydro quinacridone derivative... Specific Chemical Substances § 721.9079 Dihydro quinacridone derivative (generic). (a) Chemical substance... dihydro quinacridone derivative (PMN P-01-397) is subject to reporting under this section for the...

  5. 21 CFR 172.275 - Synthetic paraffin and succinic derivatives.

    Code of Federal Regulations, 2010 CFR

    2010-04-01

    ... 21 Food and Drugs 3 2010-04-01 2009-04-01 true Synthetic paraffin and succinic derivatives. 172... succinic derivatives. Synthetic paraffin and succinic derivatives identified in this section may be safely... acid derivatives of isopropyl alcohol, polyethylene glycol, and polypropylene glycol. It consists of a...

  6. 21 CFR 172.275 - Synthetic paraffin and succinic derivatives.

    Code of Federal Regulations, 2011 CFR

    2011-04-01

    ... 21 Food and Drugs 3 2011-04-01 2011-04-01 false Synthetic paraffin and succinic derivatives. 172... succinic derivatives. Synthetic paraffin and succinic derivatives identified in this section may be safely... acid derivatives of isopropyl alcohol, polyethylene glycol, and polypropylene glycol. It consists of a...

  7. Two-point derivative dispersion relations

    NASA Astrophysics Data System (ADS)

    Ferreira, Erasmo; Sesma, Javier

    2013-03-01

    A new derivation is given for the representation, under certain conditions, of the integral dispersion relations of scattering theory through local forms. The resulting expressions have been obtained through an independent procedure to construct the real part and consist of new mathematical structures of double infinite summations of derivatives. In this new form the derivatives are calculated at the generic value of the energy E and separately at the reference point E = m that is the lower limit of the integration. This new form may be more interesting in certain circumstances and directly shows the origin of the difficulties in convergence that were present in the old truncated forms called standard-derivative dispersion relations (DDR). For all cases in which the reductions of the double to single sums were obtained in our previous work, leading to explicit demonstration of convergence, these new expressions are seen to be identical to the previous ones. We present, as a glossary, the most simplified explicit results for the DDR's in the cases of imaginary amplitudes of forms (E/m)λ[ln (E/m)]n that cover the cases of practical interest in particle physics phenomenology at high energies. We explicitly study the expressions for the cases with λ negative odd integers, that require identification of cancelation of singularities, and provide the corresponding final results.

  8. 45 CFR 1630.12 - Applicability to derivative income.

    Code of Federal Regulations, 2010 CFR

    2010-10-01

    ..., regulations, guidelines, and instructions, the Accounting Guide for LSC recipients, the terms and conditions... 45 Public Welfare 4 2010-10-01 2010-10-01 false Applicability to derivative income. 1630.12... CORPORATION COST STANDARDS AND PROCEDURES § 1630.12 Applicability to derivative income. (a) Derivative income...

  9. Haenamindole, an unusual diketopiperazine derivative from a marine-derived Penicillium sp. KCB12F005.

    PubMed

    Kim, Jong Won; Ko, Sung-Kyun; Son, Sangkeun; Shin, Kee-Sun; Ryoo, In-Ja; Hong, Young-Soo; Oh, Hyuncheol; Hwang, Bang Yeon; Hirota, Hiroshi; Takahashi, Shunji; Kim, Bo Yeon; Osada, Hiroyuki; Jang, Jae-Hyuk; Ahn, Jong Seog

    2015-11-15

    During the chemical investigation of marine-derived fungus, an unusual diketopiperazine (DKP) alkaloid, haenamindole (1), was isolated from a culture of the marine-derived fungus Penicillium sp. KCB12F005. The structure of 1, which possesses benzyl-hydroxypiperazindione and phenyl-pyrimidoindole rings system in the molecule, was elucidated by analysis of NMR and MS data. The stereochemistry of 1 was determined by ROESY and advanced Marfey's method. Copyright © 2015 Elsevier Ltd. All rights reserved.

  10. Marine-derived angiogenesis inhibitors for cancer therapy.

    PubMed

    Wang, Ying-Qing; Miao, Ze-Hong

    2013-03-15

    Angiogenesis inhibitors have been successfully used for cancer therapy in the clinic. Many marine-derived natural products and their analogues have been reported to show antiangiogenic activities. Compared with the drugs in the clinic, these agents display interesting characteristics, including diverse sources, unique chemical structures, special modes of action, and distinct activity and toxicity profiles. This review will first provide an overview of the current marine-derived angiogenesis inhibitors based on their primary targets and/or mechanisms of action. Then, the marine-derived antiangiogenic protein kinase inhibitors will be focused on. And finally, the clinical trials of the marine-derived antiangiogenic agents will be discussed, with special emphasis on their application potentials, problems and possible coping strategies in their future development as anticancer drugs.

  11. Derivational Morphology and Base Morpheme Frequency

    ERIC Educational Resources Information Center

    Ford, M. A.; Davis, M. H.; Marslen-Wilson, W. D.

    2010-01-01

    Morpheme frequency effects for derived words (e.g. an influence of the frequency of the base "dark" on responses to "darkness") have been interpreted as evidence of morphemic representation. However, it has been suggested that most derived words would not show these effects if family size (a type frequency count claimed to reflect semantic…

  12. Carnitine derivatives: clinical usefulness.

    PubMed

    Malaguarnera, Mariano

    2012-03-01

    Carnitine and its derivatives are natural substances involved in both carbohydrate and lipid metabolism. This review summarizes the recent progress in the field in relation to the molecular mechanisms. The pool of different carnitine derivatives is formed by acetyl-L-carnitine (ALC), propionyl-L-carnitine (PLC), and isovaleryl-carnitine. ALC may have a preferential effect on the brain tissue. ALC represents a compound of great interest for its wide clinical application in various neurological disorders: it may be of benefit in treating Alzheimer's dementia, depression in the elderly, HIV infection, chronic fatigue syndrome, peripheral neuropathies, ischemia and reperfusion of the brain, and cognitive impairment associated with various conditions. PLC has been demonstrated to replenish the intermediates of the tricarboxylic acid cycle by the propionyl-CoA moiety, a greater affinity for the sarcolemmal carrier, peripheral vasodilator activity, a greater positive inotropism, and more rapid entry into myocytes. Most studies of the therapeutic use of PLC are focused on the prevention and treatment of ischemic heart disease, congestive heart failure, hypertrophic heart disease, and peripheral arterial disease. ALC and PLC are considered well tolerated without significant side-effects. A number of therapeutic effects possibly come from the interaction of carnitine and its derivatives with the elements of cellular membranes.

  13. Validated derivative and ratio derivative spectrophotometric methods for the simultaneous determination of levocetirizine dihydrochloride and ambroxol hydrochloride in pharmaceutical dosage form

    NASA Astrophysics Data System (ADS)

    Ali, Omnia I. M.; Ismail, Nahla S.; Elgohary, Rasha M.

    2016-01-01

    Three simple, precise, accurate and validated derivative spectrophotometric methods have been developed for the simultaneous determination of levocetirizine dihydrochloride (LCD) and ambroxol hydrochloride (ABH) in bulk powder and in pharmaceutical formulations. The first method is a first derivative spectrophotometric method (1D) using a zero-crossing technique of measurement at 210.4 nm for LCD and at 220.0 nm for ABH. The second method employs a second derivative spectrophotometry (2D) where the measurements were carried out at 242.0 and 224.4 nm for LCD and ABH, respectively. In the third method, the first derivative of the ratio spectra was calculated and the first derivative of the ratio amplitudes at 222.8 and 247.2 nm was selected for the determination of LCD and ABH, respectively. Calibration graphs were established in the ranges of 1.0-20.0 μg mL- 1 for LCD and 4.0-20.0 μg mL- 1 for ABH using derivative and ratio first derivative spectrophotometric methods with good correlation coefficients. The developed methods have been successfully applied to the simultaneous determination of both drugs in commercial tablet dosage form.

  14. Validated derivative and ratio derivative spectrophotometric methods for the simultaneous determination of levocetirizine dihydrochloride and ambroxol hydrochloride in pharmaceutical dosage form.

    PubMed

    Ali, Omnia I M; Ismail, Nahla S; Elgohary, Rasha M

    2016-01-15

    Three simple, precise, accurate and validated derivative spectrophotometric methods have been developed for the simultaneous determination of levocetirizine dihydrochloride (LCD) and ambroxol hydrochloride (ABH) in bulk powder and in pharmaceutical formulations. The first method is a first derivative spectrophotometric method ((1)D) using a zero-crossing technique of measurement at 210.4 nm for LCD and at 220.0 nm for ABH. The second method employs a second derivative spectrophotometry ((2)D) where the measurements were carried out at 242.0 and 224.4 nm for LCD and ABH, respectively. In the third method, the first derivative of the ratio spectra was calculated and the first derivative of the ratio amplitudes at 222.8 and 247.2 nm was selected for the determination of LCD and ABH, respectively. Calibration graphs were established in the ranges of 1.0-20.0 μg mL(-1) for LCD and 4.0-20.0 μg mL(-1) for ABH using derivative and ratio first derivative spectrophotometric methods with good correlation coefficients. The developed methods have been successfully applied to the simultaneous determination of both drugs in commercial tablet dosage form. Copyright © 2015 Elsevier B.V. All rights reserved.

  15. Dopamine-derived salsolinol derivatives as endogenous monoamine oxidase inhibitors: occurrence, metabolism and function in human brains.

    PubMed

    Naoi, Makoto; Maruyama, Wakako; Nagy, Georgy M

    2004-01-01

    Salsolinol, 1-methyl-6,7-dihydroxy-1,2,3,4-tetrahydroisoquinoline, is an endogenous catechol isoquinoline detected in humans by M. Sandler. In human brain, a series of catechol isoquinolines were identified as the condensation products of dopamine or other monoamines with aldehydes or keto-acids. Recently selective occurrence of the (R)enantiomers of salsolinol derivatives was confirmed in human brain, and they are synthesized by enzymes in situ, but not by the non-enzymatic Pictet-Spengler reaction. A (R)salsolinol synthase catalyzes the enantio-specific synthesis of (R)salsolinol from dopamine and acetaldehyde, and (R)salsolinol N-methyltransferase synthesizes N-methyl(R)salsolinol, which is further oxidized into 1,2-dimethyl-6,7-dihydroxyisoquinolinium ion by non-enzymatic and enzymatic oxidation. The step-wise reactions, N-methylation and oxidation, induce the specified distribution of the N-methylated and oxidized derivatives in the human nigro-striatum, suggesting that these derivatives may be involved in the function of dopamine neurons under physiological and pathological conditions. As shown by in vivo and in vitro experiments, salsolinol derivatives affect the levels of monoamine neurotransmitters though the inhibition of enzymes related in the metabolism of catechol- and indoleamines. In addition, the selective neurotoxicity of N-methyl(R)salsolinol to dopamine neurons was confirmed by preparation of an animal model of Parkinson's disease in rats. The involvement of N-methyl(R)salsolinol in the pathogenesis of Parkinson's disease was further indicated by the increase in the N-methyl(R)salsolinol levels in the cerebrospinal fluid and that in the activity of its synthesizing enzyme, a neural (R)salsolinol N-methyltransferase, in the lymphocytes prepared from parkinsonian patients. N-methyl(R)salsolinol induces apoptosis in dopamine neurons, which is mediated by death signal transduction in mitochondria. In addition, salsolinol was found to function as a

  16. The Basis for Photocatalytic Writing

    ERIC Educational Resources Information Center

    Ibanez, Jorge G.; Tausch, Michael W.; Bohrmann-Linde, Claudia; Fernandez-Gallardo, Isabel; Robles-Leyzaola, Ainoha; Krees, Simone; Meuter, Nico; Tennior, Mathias

    2011-01-01

    We present a demonstration involving the oxidative photobleaching of a raspberry juice dye under visible laser light irradiation using the semiconductor titanium dioxide. A plausible interpretation of the phenomenon is discussed that aids in the understanding of semiconductor energetics and the nature of light. (Contains 2 figures.)

  17. Derivation of the Freundlich Adsorption Isotherm from Kinetics

    ERIC Educational Resources Information Center

    Skopp, Joseph

    2009-01-01

    The Freundlich adsorption isotherm is a useful description of adsorption phenomena. It is frequently presented as an empirical equation with little theoretical basis. In fact, a variety of derivations exist. Here a new derivation is presented using the concepts of fractal reaction kinetics. This derivation provides an alternative basis for…

  18. Design and Synthesis of Antiblinking and Antibleaching Quantum Dots in Multiple Colors via Wave Function Confinement.

    PubMed

    Cao, Hujia; Ma, Junliang; Huang, Lin; Qin, Haiyan; Meng, Renyang; Li, Yang; Peng, Xiaogang

    2016-12-07

    Single-molecular spectroscopy reveals that photoluminescence (PL) of a single quantum dot blinks, randomly switching between bright and dim/dark states under constant photoexcitation, and quantum dots photobleach readily. These facts cast great doubts on potential applications of these promising emitters. After ∼20 years of efforts, synthesis of nonblinking quantum dots is still challenging, with nonblinking quantum dots only available in red-emitting window. Here we report synthesis of nonblinking quantum dots covering most part of the visible window using a new synthetic strategy, i.e., confining the excited-state wave functions of the core/shell quantum dots within the core quantum dot and its inner shells (≤ ∼5 monolayers). For the red-emitting ones, the new synthetic strategy yields nonblinking quantum dots with small sizes (∼8 nm in diameter) and improved nonblinking properties. These new nonblinking quantum dots are found to be antibleaching. Results further imply that the PL blinking and photobleaching of quantum dots are likely related to each other.

  19. Pulsed laser ablation of dental calculus in the near ultraviolet.

    PubMed

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

    2014-02-01

    Pulsed lasers emitting wavelengths near 400 nm can selectively ablate dental calculus without damaging underlying and surrounding sound dental hard tissue. Our results indicate that calculus ablation at this wavelength relies on the absorption of porphyrins endogenous to oral bacteria commonly found in calculus. Sub- and supragingival calculus on extracted human teeth, irradiated with 400-nm, 60-ns laser pulses at ≤8  J/cm2, exhibits a photobleached surface layer. Blue-light microscopy indicates this layer highly scatters 400-nm photons, whereas fluorescence spectroscopy indicates that bacterial porphyrins are permanently photobleached. A modified blow-off model for ablation is proposed that is based upon these observations and also reproduces our calculus ablation rates measured from laser profilometry. Tissue scattering and a stratified layering of absorbers within the calculus medium explain the gradual decrease in ablation rate from successive pulses. Depending on the calculus thickness, ablation stalling may occur at <5  J/cm2 but has not been observed above this fluence.

  20. Frapbot: An open-source application for FRAP data.

    PubMed

    Kohze, Robin; Dieteren, Cindy E J; Koopman, Werner J H; Brock, Roland; Schmidt, Samuel

    2017-08-01

    We introduce Frapbot, a free-of-charge open source software web application written in R, which provides manual and automated analyses of fluorescence recovery after photobleaching (FRAP) datasets. For automated operation, starting from data tables containing columns of time-dependent intensity values for various regions of interests within the images, a pattern recognition algorithm recognizes the relevant columns and identifies the presence or absence of prebleach values and the time point of photobleaching. Raw data, residuals, normalization, and boxplots indicating the distribution of half times of recovery (t 1/2 ) of all uploaded files are visualized instantly in a batch-wise manner using a variety of user-definable fitting options. The fitted results are provided as .zip file, which contains .csv formatted output tables. Alternatively, the user can manually control any of the options described earlier. © 2017 International Society for Advancement of Cytometry. © 2017 International Society for Advancement of Cytometry.

  1. Time multiplexing super-resolution nanoscopy based on the Brownian motion of gold nanoparticles

    NASA Astrophysics Data System (ADS)

    Ilovitsh, Tali; Ilovitsh, Asaf; Wagner, Omer; Zalevsky, Zeev

    2017-02-01

    Super-resolution localization microscopy can overcome the diffraction limit and achieve a tens of order improvement in resolution. It requires labeling the sample with fluorescent probes followed with their repeated cycles of activation and photobleaching. This work presents an alternative approach that is free from direct labeling and does not require the activation and photobleaching cycles. Fluorescently labeled gold nanoparticles in a solution are distributed on top of the sample. The nanoparticles move in a random Brownian motion, and interact with the sample. By obscuring different areas in the sample, the nanoparticles encode the sub-wavelength features. A sequence of images of the sample is captured and decoded by digital post processing to create the super-resolution image. The achievable resolution is limited by the additive noise and the size of the nanoparticles. Regular nanoparticles with diameter smaller than 100nm are barely seen in a conventional bright field microscope, thus fluorescently labeled gold nanoparticles were used, with proper

  2. Measuring Membrane Protein Dimerization Equilibrium in Lipid Bilayers by Single-Molecule Fluorescence Microscopy.

    PubMed

    Chadda, R; Robertson, J L

    2016-01-01

    Dimerization of membrane protein interfaces occurs during membrane protein folding and cell receptor signaling. Here, we summarize a method that allows for measurement of equilibrium dimerization reactions of membrane proteins in lipid bilayers, by measuring the Poisson distribution of subunit capture into liposomes by single-molecule photobleaching analysis. This strategy is grounded in the fact that given a comparable labeling efficiency, monomeric or dimeric forms of a membrane protein will give rise to distinctly different photobleaching probability distributions. These methods have been used to verify the dimer stoichiometry of the Fluc F - ion channel and the dimerization equilibrium constant of the ClC-ec1 Cl - /H + antiporter in lipid bilayers. This approach can be applied to any membrane protein system provided it can be purified, fluorescently labeled in a quantitative manner, and verified to be correctly folded by functional assays, even if the structure is not yet known. © 2016 Elsevier Inc. All rights reserved.

  3. Lateral diffusion of proteins in the periplasm of Escherichia coli.

    PubMed Central

    Brass, J M; Higgins, C F; Foley, M; Rugman, P A; Birmingham, J; Garland, P B

    1986-01-01

    We have introduced biologically active, fluorescently labeled maltose-binding protein into the periplasmic space of Escherichia coli and measured its lateral diffusion coefficient by the fluorescence photobleaching recovery method. Diffusion of this protein in the periplasm was found to be surprisingly low (lateral diffusion coefficient, 0.9 X 10(-10) cm2 s-1), about 1,000-fold lower than would be expected for diffusion in aqueous medium and almost 100-fold lower than for an equivalent-size protein in the cytoplasm. Galactose-binding protein, myoglobin, and cytochrome c were also introduced into the periplasm and had diffusion coefficients identical to that determined for the maltose-binding protein. For all proteins nearly 100% recovery of fluorescence was obtained after photobleaching, indicating that the periplasm is a single contiguous compartment surrounding the cell. These data have considerable implications for periplasmic structure and for the role of periplasmic proteins in transport and chemotaxis. Images PMID:3005237

  4. Hydroxyurea derivatives of irofulven with improved antitumor efficacy.

    PubMed

    Staake, Michael D; Kashinatham, Alisala; McMorris, Trevor C; Estes, Leita A; Kelner, Michael J

    2016-04-01

    Irofulven is a semi-synthetic derivative of Illudin S, a toxic sesquiterpene isolated from the mushroom Omphalotus illudens. Irofulven has displayed significant antitumor activity in various clinical trials but displayed a limited therapeutic index. A new derivative of irofulven was prepared by reacting hydroxyurea with irofulven under acidic conditions. Acetylation of this new compound with acetic anhydride produced a second derivative. Both of these new derivatives displayed significant antitumor activity in vitro and in vivo comparable to or exceeding that of irofulven. Copyright © 2016 Elsevier Ltd. All rights reserved.

  5. Adult Langerhans cells derive predominantly from embryonic fetal liver monocytes with a minor contribution of yolk sac-derived macrophages.

    PubMed

    Hoeffel, Guillaume; Wang, Yilin; Greter, Melanie; See, Peter; Teo, Pearline; Malleret, Benoit; Leboeuf, Marylène; Low, Donovan; Oller, Guillaume; Almeida, Francisca; Choy, Sharon H Y; Grisotto, Marcos; Renia, Laurent; Conway, Simon J; Stanley, E Richard; Chan, Jerry K Y; Ng, Lai Guan; Samokhvalov, Igor M; Merad, Miriam; Ginhoux, Florent

    2012-06-04

    Langerhans cells (LCs) are the dendritic cells (DCs) of the epidermis, forming one of the first hematopoietic lines of defense against skin pathogens. In contrast to other DCs, LCs arise from hematopoietic precursors that seed the skin before birth. However, the origin of these embryonic precursors remains unclear. Using in vivo lineage tracing, we identify a first wave of yolk sac (YS)-derived primitive myeloid progenitors that seed the skin before the onset of fetal liver hematopoiesis. YS progenitors migrate to the embryo proper, including the prospective skin, where they give rise to LC precursors, and the brain rudiment, where they give rise to microglial cells. However, in contrast to microglia, which remain of YS origin throughout life, YS-derived LC precursors are largely replaced by fetal liver monocytes during late embryogenesis. Consequently, adult LCs derive predominantly from fetal liver monocyte-derived cells with a minor contribution of YS-derived cells. Altogether, we establish that adult LCs have a dual origin, bridging early embryonic and late fetal myeloid development.

  6. Adult Langerhans cells derive predominantly from embryonic fetal liver monocytes with a minor contribution of yolk sac–derived macrophages

    PubMed Central

    Hoeffel, Guillaume; Wang, Yilin; Greter, Melanie; See, Peter; Teo, Pearline; Malleret, Benoit; Leboeuf, Marylène; Low, Donovan; Oller, Guillaume; Almeida, Francisca; Choy, Sharon H.Y.; Grisotto, Marcos; Renia, Laurent; Conway, Simon J.; Stanley, E. Richard; Chan, Jerry K.Y.; Ng, Lai Guan; Samokhvalov, Igor M.

    2012-01-01

    Langerhans cells (LCs) are the dendritic cells (DCs) of the epidermis, forming one of the first hematopoietic lines of defense against skin pathogens. In contrast to other DCs, LCs arise from hematopoietic precursors that seed the skin before birth. However, the origin of these embryonic precursors remains unclear. Using in vivo lineage tracing, we identify a first wave of yolk sac (YS)–derived primitive myeloid progenitors that seed the skin before the onset of fetal liver hematopoiesis. YS progenitors migrate to the embryo proper, including the prospective skin, where they give rise to LC precursors, and the brain rudiment, where they give rise to microglial cells. However, in contrast to microglia, which remain of YS origin throughout life, YS-derived LC precursors are largely replaced by fetal liver monocytes during late embryogenesis. Consequently, adult LCs derive predominantly from fetal liver monocyte-derived cells with a minor contribution of YS-derived cells. Altogether, we establish that adult LCs have a dual origin, bridging early embryonic and late fetal myeloid development. PMID:22565823

  7. Polyimides Derived from Novel Asymmetric Benzophenone Dianhydrides

    NASA Technical Reports Server (NTRS)

    Chuang, Chun-Hua (Inventor)

    2015-01-01

    This invention relates to the composition and processes for preparing thermoset polyimides derived from an asymmetric dianhydride, namely 2,3,3',4'-benzophenone dianhydride (a-BTDA) with at least one diamine, and a monofunctional terminal endcaps. The monofunctional terminating groups include 4-phenylethynylphthalic anhydride ester-acid derivatives, phenylethyl trimellitic anhydride (PETA) and its ester derivatives as well as 3-phenylethynylaniline. The process of polyimide composite comprises impregnating monomer reactants of dianhydride or its ester-acid derivatives, diamine and with monofunctional reactive endcaps into glass, carbon, quartz or synthetic fibers and fabrics, and then stack up into laminates and subsequently heated to between 150-375.degree. C. either at atmosphere or under pressure to promote the curing and crosslinking of the reactive endcaps to form a network of thermoset polyimides.

  8. 21 CFR 184.1317 - Garlic and its derivatives.

    Code of Federal Regulations, 2011 CFR

    2011-04-01

    ... 21 Food and Drugs 3 2011-04-01 2011-04-01 false Garlic and its derivatives. 184.1317 Section 184... Listing of Specific Substances Affirmed as GRAS § 184.1317 Garlic and its derivatives. (a) Garlic is the... derivatives include essential oils, oleo-resins, and natural extractives obtained from garlic. (b) Garlic oil...

  9. Generalized Second-Order Partial Derivatives of 1/r

    ERIC Educational Resources Information Center

    Hnizdo, V.

    2011-01-01

    The generalized second-order partial derivatives of 1/r, where r is the radial distance in three dimensions (3D), are obtained using a result of the potential theory of classical analysis. Some non-spherical-regularization alternatives to the standard spherical-regularization expression for the derivatives are derived. The utility of a…

  10. Derivation and definition of a linear aircraft model

    NASA Technical Reports Server (NTRS)

    Duke, Eugene L.; Antoniewicz, Robert F.; Krambeer, Keith D.

    1988-01-01

    A linear aircraft model for a rigid aircraft of constant mass flying over a flat, nonrotating earth is derived and defined. The derivation makes no assumptions of reference trajectory or vehicle symmetry. The linear system equations are derived and evaluated along a general trajectory and include both aircraft dynamics and observation variables.

  11. Preparation and Characterization of Fluorescent Derivatives of Lysozyme

    NASA Technical Reports Server (NTRS)

    Smith, Lori; Pusey, Marc

    1998-01-01

    Fluorescence is one of the most versatile and powerful tools for the study of macromolecules. However, its use in macromolecular crystal growth studies is hampered by the necessity of preparing fluorescent derivatives where the probe does not markedly affect the crystal packing. Alternatively, one can prepare derivatives of limited utility if it is known that they will not affect the specific goals of a given study. We have prepared a number of fluorescent derivatives of chicken egg white lysozyme, covalently attaching fluorescent probes to two different sites on the protein molecule. The first site is the side chain carboxyl group of ASP 101. Amine containing probes such as lucifer yellow, cascade blue, and 5- (2-aminoethyl) aminonapthalene-l-sulfonic acid (EDANS) have been attached using a carbodiimide coupling procedure. ASP 101 lies within the active site cleft, and it is believed that the probes are "buried" within that cleft. This is supported by the fact that all such derivatives have been found to crystallize, with the crystals being fluorescent. Tetragonal crystals of the lucifer yellow derivative have been found to diffract to at least 1.9 A resolution. X-ray diffraction data has been acquired and we are now working on the structure of this derivative. The second group of derivatives is to the N-terminal amine group. The derivatization reaction is performed by using a succinimidyl ester of the probe to be attached. Fluorescent probes such as pyrene acetic acid, 5-carboxyfluorescein, and Oregon green have been attached to this site. We have had little success in crystallizing these derivatives, probably because this site is part of the contact region between the 43 helix chains. However, these sites do not interfere with formation of the 43 helices and the derivatives are suitable for study of their formation in solution. The derivatives are being characterized by steady state and lifetime fluorescence methods, and the presentation will discuss these

  12. Paracrine Engineering of Human Explant-Derived Cardiac Stem Cells to Over-Express Stromal-Cell Derived Factor 1α Enhances Myocardial Repair.

    PubMed

    Tilokee, Everad L; Latham, Nicholas; Jackson, Robyn; Mayfield, Audrey E; Ye, Bin; Mount, Seth; Lam, Buu-Khanh; Suuronen, Erik J; Ruel, Marc; Stewart, Duncan J; Davis, Darryl R

    2016-07-01

    First generation cardiac stem cell products provide indirect cardiac repair but variably produce key cardioprotective cytokines, such as stromal-cell derived factor 1α, which opens the prospect of maximizing up-front paracrine-mediated repair. The mesenchymal subpopulation within explant derived human cardiac stem cells underwent lentiviral mediated gene transfer of stromal-cell derived factor 1α. Unlike previous unsuccessful attempts to increase efficacy by boosting the paracrine signature of cardiac stem cells, cytokine profiling revealed that stromal-cell derived factor 1α over-expression prevented lv-mediated "loss of cytokines" through autocrine stimulation of CXCR4+ cardiac stem cells. Stromal-cell derived factor 1α enhanced angiogenesis and stem cell recruitment while priming cardiac stem cells to readily adopt a cardiac identity. As compared to injection with unmodified cardiac stem cells, transplant of stromal-cell derived factor 1α enhanced cells into immunodeficient mice improved myocardial function and angiogenesis while reducing scarring. Increases in myocardial stromal-cell derived factor 1α content paralleled reductions in myocyte apoptosis but did not influence long-term engraftment or the fate of transplanted cells. Transplantation of stromal-cell derived factor 1α transduced cardiac stem cells increased the generation of new myocytes, recruitment of bone marrow cells, new myocyte/vessel formation and the salvage of reversibly damaged myocardium to enhance cardiac repair after experimental infarction. Stem Cells 2016;34:1826-1835. © 2016 AlphaMed Press.

  13. Synthesis and characterization of citrate-based fluorescent small molecules and biodegradable polymers.

    PubMed

    Xie, Zhiwei; Kim, Jimin P; Cai, Qing; Zhang, Yi; Guo, Jinshan; Dhami, Ranjodh S; Li, Li; Kong, Bin; Su, Yixue; Schug, Kevin A; Yang, Jian

    2017-03-01

    Novel citric acid based photoluminescent dyes and biodegradable polymers are synthesized via a facile "one-pot" reaction. A comprehensive understanding of the fluorescence mechanisms of the resulting citric acid-based fluorophores is reported. Two distinct types of fluorophores are identified: a thiozolopyridine family with high quantum yield, long lifetime, and exceptional photostability, and a dioxopyridine family with relatively lower quantum yield, multiple lifetimes, and solvent-dependent band shifting behavior. Applications in molecular labeling and cell imaging were demonstrated. The above discoveries contribute to the field of fluorescence chemistry and have laid a solid foundation for further development of new fluorophores and materials that show promise in a diversity of fluorescence-based applications. Photoluminescent materials are pivotal for fluorescence based imaging, labeling and sensing applications. Understanding their fluorescence mechanism is challenging and imperative. We develop a new class of citric acid-derived fluorescent materials in forms of polymers and small molecular dyes by a one-step solvent free reaction. We discovered two different classes of citric acid-derived fluorophores. A two-ring thiozolopyridine structure demonstrates strong fluorescence and exceptional resistance to photo-bleaching. A one-ring dioxopyridine exhibits relative weak fluorescence but with intriguing excitation and solvent-dependent emission wavelength shifting. Our methodology of synthesizing citric acid-derived fluorophores and the understanding on their luminescence are instrumental to the design and production of a large number of new photoluminescent materials for biological and biomedical applications. Copyright © 2017 Acta Materialia Inc. Published by Elsevier Ltd. All rights reserved.

  14. Modelling ocean-colour-derived chlorophyll a

    NASA Astrophysics Data System (ADS)

    Dutkiewicz, Stephanie; Hickman, Anna E.; Jahn, Oliver

    2018-01-01

    This article provides a proof of concept for using a biogeochemical/ecosystem/optical model with a radiative transfer component as a laboratory to explore aspects of ocean colour. We focus here on the satellite ocean colour chlorophyll a (Chl a) product provided by the often-used blue/green reflectance ratio algorithm. The model produces output that can be compared directly to the real-world ocean colour remotely sensed reflectance. This model output can then be used to produce an ocean colour satellite-like Chl a product using an algorithm linking the blue versus green reflectance similar to that used for the real world. Given that the model includes complete knowledge of the (model) water constituents, optics and reflectance, we can explore uncertainties and their causes in this proxy for Chl a (called derived Chl a in this paper). We compare the derived Chl a to the actual model Chl a field. In the model we find that the mean absolute bias due to the algorithm is 22 % between derived and actual Chl a. The real-world algorithm is found using concurrent in situ measurement of Chl a and radiometry. We ask whether increased in situ measurements to train the algorithm would improve the algorithm, and find a mixed result. There is a global overall improvement, but at the expense of some regions, especially in lower latitudes where the biases increase. Not surprisingly, we find that region-specific algorithms provide a significant improvement, at least in the annual mean. However, in the model, we find that no matter how the algorithm coefficients are found there can be a temporal mismatch between the derived Chl a and the actual Chl a. These mismatches stem from temporal decoupling between Chl a and other optically important water constituents (such as coloured dissolved organic matter and detrital matter). The degree of decoupling differs regionally and over time. For example, in many highly seasonal regions, the timing of initiation and peak of the

  15. New well testing applications of the pressure derivative

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

    Onur, M.

    1989-01-01

    This work presents new derivative type curves based on a new derivative group which is equal to the dimensionless pressure group divided by its logarithmic derivative with respect to dimensionless time group. One major advantage of these type curves is that the type-curve match of field pressure/pressure-derivative data with the new derivative type curves is accomplished by moving the field data plot in only the horizontal direction. This type-curve match fixes time match-point values. The pressure change versus time data is then matched with the dimensionless pressure solution to determine match-point values. Well/reservoir parameters can then be estimated in themore » standard way. This two step type-curve matching procedure increases the likelihood of obtaining a unique match. Moreover, the unique correspondence between the ordinate of the field data plot and the new derivative type curves should prove useful in determining whether given field data actually represents the well/reservoir model assumed by a selected type curve solution. It is also shown that the basic idea used in construction the type curves can be used to ensure that proper semilog straight lines are chosen when analyzing pressure data by semilog methods. Analysis of both drawdown and buildup data is considered and actual field cases are analyzed using the new derivative type curves and the semilog identification method. This work also presents new methods based on the pressure derivative to analyze buildup data obtained at a well (fracture or unfractured) produced to pseudosteady-state prior to shut-in. By using a method of analysis based on the pressure derivative, it is shown that a well's drainage area at the instant of shut-in and the flow capacity can be computed directly from buildup data even in cases where conventional semilog straight lines are not well-defined.« less

  16. Deriving Case, Agreement and Voice Phenomena in Syntax

    ERIC Educational Resources Information Center

    Sigurdsson, Einar Freyr

    2017-01-01

    This dissertation places case, agreement and Voice phenomena in syntax. It argues that the derivation is driven by so-called derivational features, that is, structure-building features (Merge) and probe features (Agree) (Heck and Muller 2007 and Muller 2010; see also Chomsky 2000, 2001). Both types are essential in deriving case and agreement in…

  17. 17 CFR 240.16b-6 - Derivative securities.

    Code of Federal Regulations, 2010 CFR

    2010-04-01

    ... 17 Commodity and Securities Exchanges 3 2010-04-01 2010-04-01 false Derivative securities. 240.16b... Exchange Act of 1934 Exemption of Certain Transactions from Section 16(b) § 240.16b-6 Derivative securities...). Note to paragraph (b): The exercise or conversion of a derivative security that does not satisfy the...

  18. Correlation of Predicted and Flight Derived Stability and Control Derivatives with Particular Application to Tailless Delta Wing Configurations

    NASA Technical Reports Server (NTRS)

    Weil, J.

    1981-01-01

    Flight derived longitudinal and lateral-directional stability and control derivatives were compared to wind-tunnel derived values. As a result of these comparisons, boundaries representing the uncertainties that could be expected from wind-tunnel predictions were established. These boundaries provide a useful guide for control system sensitivity studies prior to flight. The primary application for this data was the space shuttle, and as a result the configurations included in the study were those most applicable to the space shuttle. The configurations included conventional delta wing aircraft as well as the X-15 and lifting body vehicles.

  19. Effects of sulfate chitosan derivatives on nonalcoholic fatty liver disease

    NASA Astrophysics Data System (ADS)

    Yu, Mingming; Wang, Yuanhong; Jiang, Tingfu; Lv, Zhihua

    2014-06-01

    Sulfate chitosan derivatives have good solubility and therapeutic effect on the cell model of NAFLD. The aim of this study was to examine the therapeutic effect of sulfate chitosan derivatives on NAFLD. The male Wistar rats were orally fed high fat emulsion and received sulfate chitosan derivatives for 5 weeks to determine the pre-treatment effect of sulfate chitosan derivatives on NAFLD. To evaluate the therapeutic effect of sulfate chitosan derivatives on NAFLD, the rats were orally fed with high concentration emulsion for 5 weeks, followed by sulfate chitosan derivatives for 3 weeks. Histological analysis and biomedical assays showed that sulfate chitosan derivatives can dramatically prevent the development of hepatic steatosis in hepatocyte cells. In animal studies, pre-treatment and treatment with sulfate chitosan derivatives significantly protected against hepatic steatohepatitis induced by high fat diet according to histological analysis. Furthermore, increased TC, ALT, MDA, and LEP in NAFLD were significantly ameliorated by pre-treatment and treatment with sulfate chitosan derivatives. Furthermore, increased TG, AST, and TNF-α in NAFLD were significantly ameliorated by treatment with sulfate chitosan derivatives. Sulfate chitosan derivatives have good pre-treatment and therapeutic effect on NAFLD.

  20. Crystallisation and crystal forms of carbohydrate derivatives

    NASA Astrophysics Data System (ADS)

    Lennon, Lorna

    This thesis is focused on the synthesis and solid state analysis of carbohydrate derivatives, including many novel compounds. Although the synthetic chemistry surrounding carbohydrates is well established in the literature, the crystal chemistry of carbohydrates is less well studied. Therefore this research aims to improve understanding of the solid state properties of carbohydrate derivatives through gaining more information on their supramolecular bonding. Chapter One focuses on an introduction to the solid state of organic compounds, with a background to crystallisation, including issues that can arise during crystal growth. Chapter Two is based on glucopyranuronate derivatives which are understudied in terms of their solid state forms. This chapter reports on the formation of novel glucuronamides and utilising the functionality of the amide bond for crystallisation. TEMPO oxidation was completed to form glucopyranuronates by oxidation of the primary alcohol groups of glucosides to the carboxylic acid derivatives, to increase functionality for enhanced crystal growth. Chapter Three reports on the synthesis of glucopyranoside derivatives by O-glycosylation reactions and displays crystal structures, including a number of previously unsolved acetate protected and deprotected crystal structures. More complex glycoside derivatives were also researched in an aim to study the resultant supramolecular motifs. Chapter Four contains the synthesis of aryl cellobioside derivatives including the novel crystal structures that were solved for the acetate protected and deprotected compounds. Research was carried out to determine if 1-deoxycellodextrins could act as putative isostructures for cellulose. Our research displays the presence of isostructural references with 1-deoxycellotriose shown to be similar to cellulose III11, 1-deoxycellotetraose correlates with cellulose IV11 and 1-deoxycellopentose shows isostructurality similar to that of cellulose II. Chapter Five contains

  1. Synthesis of Novel Basic Skeletons Derived from Naltrexone

    NASA Astrophysics Data System (ADS)

    Nagase, Hiroshi; Fujii, Hideaki

    We will describe eight interesting reactions using naltrexone derivatives. Almost all these reactions are characteristic of naltrexone derivatives, and can lead to the synthesis of many novel skeletons that provide new interesting pharmacological data. Some of the new reactions that were found with naltrexone derivatives were expanded into general reactions. For example, the reaction of 6α-hydroxyaldehyde derived from naltrexone led to the oxazoline dimer and the 1,3,5-trioxazatriquinane skeleton (triplet drug); this reaction was applied to general ketones which were converted to α-hydroxyaldehydes, followed by conversion to dimers and trimers, as described in Sect. 7.

  2. Derivation of GFDM Based on OFDM Principles

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

    Hussein Moradi; Behrouz Farhang-Boroujeny

    2015-06-01

    This paper starts with discussing the principle based on which the celebrated orthogonal frequency division multiplexing (OFDM) signals are constructed. It then extends the same principle to construct the newly introduced generalized frequency division multiplexing (GFDM) signals. This novel derivation sheds light on some interesting properties of GFDM. In particular, our derivation seamlessly leads to an implementation of GFDM transmitter which has significantly lower complexity than what has been reported so far. Our derivation also facilitates a trivial understanding of how GFDM (similar to OFDM) can be applied in MIMO channels.

  3. Ineffective higher derivative black hole hair

    NASA Astrophysics Data System (ADS)

    Goldstein, Kevin; Mashiyane, James Junior

    2018-01-01

    Inspired by the possibility that the Schwarzschild black hole may not be the unique spherically symmetric vacuum solution to generalizations of general relativity, we consider black holes in pure fourth order higher derivative gravity treated as an effective theory. Such solutions may be of interest in addressing the issue of higher derivative hair or during the later stages of black hole evaporation. Non-Schwarzschild solutions have been studied but we have put earlier results on a firmer footing by finding a systematic asymptotic expansion for the black holes and matching them with known numerical solutions obtained by integrating out from the near-horizon region. These asymptotic expansions can be cast in the form of trans-series expansions which we conjecture will be a generic feature of non-Schwarzschild higher derivative black holes. Excitingly we find a new branch of solutions with lower free energy than the Schwarzschild solution, but as found in earlier work, solutions only seem to exist for black holes with large curvatures, meaning that one should not generically neglect even higher derivative corrections. This suggests that one effectively recovers the nonhair theorems in this context.

  4. Ferroquine and its derivatives: new generation of antimalarial agents.

    PubMed

    Wani, Waseem A; Jameel, Ehtesham; Baig, Umair; Mumtazuddin, Syed; Hun, Lee Ting

    2015-08-28

    Malaria has been teasing human populations from a long time. Presently, several classes of antimalarial drugs are available in market, but the issues of toxicity, lower efficacy and the resistance by malarial parasites have decreased their overall therapeutic indices. Thus, the search for new promising antimalarials continues, however, the battle against malaria is far from over. Ferroquine is a derivative of chloroquine with antimalarial properties. It is the most successful of the chloroquine derivatives. Not only ferroquine, but also its derivatives have shown promising potential as antimalarials of clinical interest. Presently, much research is dedicated to the development of ferroquine derivatives as safe alternatives to antimalarial chemotherapy. The present article describes the structural, chemical and biological features of ferroquine. Several classes of ferroquine derivatives including hydroxyferroquines, trioxaferroquines, chloroquine-bridged ferrocenophanes, thiosemicarbazone derivatives, ferrocene dual conjugates, 4-N-substituted derivatives, and others have been discussed. Besides, the mechanism of action of ferroquine has been discussed. A careful observation has been made into pharmacologically significant ferroquine derivatives with better or equal therapeutic effects to that of chloroquine and ferroquine. A brief discussion of the toxicities of ferroquine derivatives has been made. Finally, efforts have been made to discuss the current challenges and future perspectives of ferroquine-based antimalarial drug development. Copyright © 2015 Elsevier Masson SAS. All rights reserved.

  5. Systemic T Cells Immunosuppression of Glioma Stem Cell-Derived Exosomes Is Mediated by Monocytic Myeloid-Derived Suppressor Cells

    PubMed Central

    Domenis, Rossana; Cesselli, Daniela; Toffoletto, Barbara; Bourkoula, Evgenia; Caponnetto, Federica; Manini, Ivana; Beltrami, Antonio Paolo; Ius, Tamara; Skrap, Miran; Di Loreto, Carla

    2017-01-01

    A major contributing factor to glioma development and progression is its ability to evade the immune system. Nano-meter sized vesicles, exosomes, secreted by glioma-stem cells (GSC) can act as mediators of intercellular communication to promote tumor immune escape. Here, we investigated the immunomodulatory properties of GCS-derived exosomes on different peripheral immune cell populations. Healthy donor peripheral blood mononuclear cells (PBMCs) stimulated with anti-CD3, anti-CD28 and IL-2, were treated with GSC-derived exosomes. Phenotypic characterization, cell proliferation, Th1/Th2 cytokine secretion and intracellular cytokine production were analysed by distinguishing among effector T cells, regulatory T cells and monocytes. In unfractionated PBMCs, GSC-derived exosomes inhibited T cell activation (CD25 and CD69 expression), proliferation and Th1 cytokine production, and did not affect cell viability or regulatory T-cell suppression ability. Furthermore, exosomes were able to enhance proliferation of purified CD4+ T cells. In PBMCs culture, glioma-derived exosomes directly promoted IL-10 and arginase-1 production and downregulation of HLA-DR by unstimulated CD14+ monocytic cells, that displayed an immunophenotype resembling that of monocytic myeloid-derived suppressor cells (Mo-MDSCs). Importantly, the removal of CD14+ monocytic cell fraction from PBMCs restored T-cell proliferation. The same results were observed with exosomes purified from plasma of glioblastoma patients. Our results indicate that glioma-derived exosomes suppress T-cell immune response by acting on monocyte maturation rather than on direct interaction with T cells. Selective targeting of Mo-MDSC to treat glioma should be considered with regard to how immune cells allow the acquirement of effector functions and therefore counteracting tumor progression. PMID:28107450

  6. Systemic T Cells Immunosuppression of Glioma Stem Cell-Derived Exosomes Is Mediated by Monocytic Myeloid-Derived Suppressor Cells.

    PubMed

    Domenis, Rossana; Cesselli, Daniela; Toffoletto, Barbara; Bourkoula, Evgenia; Caponnetto, Federica; Manini, Ivana; Beltrami, Antonio Paolo; Ius, Tamara; Skrap, Miran; Di Loreto, Carla; Gri, Giorgia

    2017-01-01

    A major contributing factor to glioma development and progression is its ability to evade the immune system. Nano-meter sized vesicles, exosomes, secreted by glioma-stem cells (GSC) can act as mediators of intercellular communication to promote tumor immune escape. Here, we investigated the immunomodulatory properties of GCS-derived exosomes on different peripheral immune cell populations. Healthy donor peripheral blood mononuclear cells (PBMCs) stimulated with anti-CD3, anti-CD28 and IL-2, were treated with GSC-derived exosomes. Phenotypic characterization, cell proliferation, Th1/Th2 cytokine secretion and intracellular cytokine production were analysed by distinguishing among effector T cells, regulatory T cells and monocytes. In unfractionated PBMCs, GSC-derived exosomes inhibited T cell activation (CD25 and CD69 expression), proliferation and Th1 cytokine production, and did not affect cell viability or regulatory T-cell suppression ability. Furthermore, exosomes were able to enhance proliferation of purified CD4+ T cells. In PBMCs culture, glioma-derived exosomes directly promoted IL-10 and arginase-1 production and downregulation of HLA-DR by unstimulated CD14+ monocytic cells, that displayed an immunophenotype resembling that of monocytic myeloid-derived suppressor cells (Mo-MDSCs). Importantly, the removal of CD14+ monocytic cell fraction from PBMCs restored T-cell proliferation. The same results were observed with exosomes purified from plasma of glioblastoma patients. Our results indicate that glioma-derived exosomes suppress T-cell immune response by acting on monocyte maturation rather than on direct interaction with T cells. Selective targeting of Mo-MDSC to treat glioma should be considered with regard to how immune cells allow the acquirement of effector functions and therefore counteracting tumor progression.

  7. Thymol derivatives from hairy roots of Arnica montana.

    PubMed

    Weremczuk-Jezyna, I; Kisiel, W; Wysokińska, H

    2006-09-01

    Five known thymol derivatives were isolated from roots of Arnica montana transformed with Agrobacterium rhizogenes LBA 9402. The compounds were characterized by spectral methods. The pattern of thymol derivatives in light-grown hairy roots was slightly different from that in dark-grown ones. This is the first report on the presence of thymol derivatives in hairy roots of the plant.

  8. Secondary metabolites from marine-derived microorganisms.

    PubMed

    Chen, Gang; Wang, Hai-Feng; Pei, Yue-Hu

    2014-01-01

    In the search for novel and bioactive molecules for drug discovery, marine-derived natural resources, especially marine microorganisms are becoming an important and interesting research area. This study covers the literature published after 2008 on secondary metabolites of marine-derived microorganisms. The emphasis was on new compounds with the relevant biological activities, strain information, and country of origin. New compounds without biological activity were not included.

  9. Pyrazole derivatives as antitumor, anti-inflammatory and antibacterial agents.

    PubMed

    Liu, Jia-Jia; Zhao, Meng-Yue; Zhang, Xin; Zhao, Xin; Zhu, Hai-Liang

    2013-11-01

    Within the past years, many researches on the synthesis, structure-activity relationships (SAR), antitumor, antiinflammatory and anti-bacterial activities of the pyrazole derivatives have been reported. Several pyrazole derivatives possess important pharmacological activities and they have been proved useful materials in drug research. Pyrazole derivatives play an important role in antitumor agents because of their good inhibitory activity against BRAF(V600E), EGFR, telomerase, ROS Receptor Tyrosine Kinase and Aurora-A kinase. In addition, pyrazole derivatives also show good antiinflammatory and anti-bacterial activities. In this review, the bioactivities of the pyrazole derivatives mentioned above will be summarized in detail. We sincerely hope that increasing knowledge of the SAR and cellular processes underlying the bioactivity of pyrazole derivatives will be beneficial to the rational design of new generation of small molecule drugs.

  10. Mass Spectrometric Characteristics of Prenylated Indole Derivatives from Marine-Derived Penicillium sp. NH-SL

    PubMed Central

    Ding, Hui; Ding, Wanjing; Ma, Zhongjun

    2017-01-01

    Two prenylated indole alkaloids were isolated from the ethyl acetate extracts of a marine-derived fungus Penicillium sp. NH-SL and one of them exhibited potent cytotoxic activity against mouse hepa 1c1c7 cells. In order to detect other bioactive analogs, we used liquid chromatogram tandem mass spectrometry (LC-MS/MS) to analyze the mass spectrometric characteristics of the isolated compounds as well as the crude extracts. As a result, three other analogs were detected, and their structures were deduced according to the similar fragmentation patterns. This is the first systematic report on the mass spectrometric characteristics of prenylated indole derivatives. PMID:28327529

  11. Mass Spectrometric Characteristics of Prenylated Indole Derivatives from Marine-Derived Penicillium sp. NH-SL.

    PubMed

    Ding, Hui; Ding, Wanjing; Ma, Zhongjun

    2017-03-22

    Two prenylated indole alkaloids were isolated from the ethyl acetate extracts of a marine-derived fungus Penicillium sp. NH-SL and one of them exhibited potent cytotoxic activity against mouse hepa 1c1c7 cells. In order to detect other bioactive analogs, we used liquid chromatogram tandem mass spectrometry (LC-MS/MS) to analyze the mass spectrometric characteristics of the isolated compounds as well as the crude extracts. As a result, three other analogs were detected, and their structures were deduced according to the similar fragmentation patterns. This is the first systematic report on the mass spectrometric characteristics of prenylated indole derivatives.

  12. Redistribution of fluorescently labeled tubulin in the mitotic apparatus of sand dollar eggs and the effects of taxol.

    PubMed

    Hamaguchi, Y; Toriyama, M; Sakai, H; Hiramoto, Y

    1987-02-01

    Fluorescently labeled tubulin was quickly incorporated into the mitotic apparatus when injected into a live sand dollar egg. After a rectangular area (1.6 X 16 microns) of the mitotic spindle was photobleached at metaphase or anaphase by the irradiation of a laser microbeam, redistribution of fluorescence was almost complete within 30 sec. The photobleached area did not change in shape during the redistribution. During the period of redistribution, the bleached area moved slightly toward the near pole at metaphase and anaphase (means: 1.6 and 1.8 micron/min, respectively). These results indicate that redistribution was not due to the exchange of tubulin subunits only at the ends of microtubules but to their rapid exchange at sites along the microtubules in the bleached region. Furthermore, treadmilling of tubulin molecules along with the spindle microtubules possibly occurred at the rate of 1.6 micron/min at metaphase. Birefringence of the mitotic apparatus increased with a large increase in both the number and length of astral rays shortly after taxol was injected. However, the microtubules did not all seem to elongate at the same rate but appeared to become equalized in length. Chromosome movement stopped within 60 sec after the injection. Centrospheres became large and the labeled tubulin already incorporated into the centrospheres was excluded from the enlarged centrospheres. Shortly after the labeled tubulin was injected following the injection of taxol, it accumulated in the peripheral region of the centrospheres, suggesting that microtubules first assembled at this region. Fluorescently labeled tubulin in the mitotic apparatus in the egg after injection of taxol was redistributed much more slowly after photobleaching than in uninjected eggs.

  13. Possibility for a full optical determination of photodynamic therapy outcome

    NASA Astrophysics Data System (ADS)

    Vollet-Filho, J. D.; Menezes, P. F. C.; Moriyama, L. T.; Grecco, C.; Sibata, C.; Allison, R. R.; Castro e Silva, O.; Bagnato, V. S.

    2009-05-01

    The efficacy of photodynamic therapy (PDT) depends on a variety of parameters: concentration of the photosensitizer at the time of treatment, light wavelength, fluence, fluence rate, availability of oxygen within the illuminated volume, and light distribution in the tissue. Dosimetry in PDT requires the congregation of adequate amounts of light, drug, and tissue oxygen. The adequate dosimetry should be able to predict the extension of the tissue damage. Photosensitizer photobleaching rate depends on the availability of molecular oxygen in the tissue. Based on photosensitizers photobleaching models, high photobleaching has to be associated with high production of singlet oxygen and therefore with higher photodynamic action, resulting in a greater depth of necrosis. The purpose of this work is to show a possible correlation between depth of necrosis and the in vivo photosensitizer (in this case, Photogem®) photodegradation during PDT. Such correlation allows possibilities for the development of a real time evaluation of the photodynamic action during PDT application. Experiments were performed in a range of fluence (0-450 J/cm2) at a constant fluence rate of 250 mW/cm2 and applying different illumination times (0-1800 s) to achieve the desired fluence. A quantity was defined (ψ) as the product of fluorescence ratio (related to the photosensitizer degradation at the surface) and the observed depth of necrosis. The correlation between depth of necrosis and surface fluorescence signal is expressed in ψ and could allow, in principle, a noninvasive monitoring of PDT effects during treatment. High degree of correlation is observed and a simple mathematical model to justify the results is presented.

  14. Serotonergic ergoline derivatives.

    PubMed

    Mantegani, S; Brambilla, E; Caccia, C; Damiani, G; Fornaretto, M G; McArthur, R A; Varasi, M

    1998-05-05

    Novel classes of 13- and 14-tertbutyl-ergoline derivatives were prepared, and characterised in vitro for their affinity for adrenergic, dopaminergic and serotonergic binding sites. This study particularly examines the importance of the presence and the position of the tert-butyl group in conferring either significant 5-HT1A or 5-HT2 affinity and selectivity respectively.

  15. Development of an accurate and high-throughput methodology for structural comprehension of chlorophylls derivatives. (II) Dephytylated derivatives.

    PubMed

    Chen, Kewei; Ríos, José Julián; Roca, María; Pérez-Gálvez, Antonio

    2015-09-18

    Dephytylated chlorophylls (chlorophyllides and pheophorbides) are the starting point of the chlorophyll catabolism in green tissues, components of the chlorophyll pattern in storage/processed food vegetables, as well as the favoured structural arrangement for chlorophyll absorption. In addition, dephytylated native chlorophylls are prone to several modifications of their structure yielding pyro-, 13(2)-hydroxy- and 15(1)-hydroxy-lactone derivatives. Despite of these outstanding remarks only few of them have been analysed by MS(n). Besides new protocols for obtaining standards, we have developed a new high throughput methodology able to determine the fragmentation pathway of 16 dephytylated chlorophyll derivatives, elucidating the structures of the new product ions and new mechanisms of fragmentation. The new methodology combines, by first time, high resolution time-of-flight mass spectrometry and powerful post-processing software. Native chlorophyllides and pheophorbides mainly exhibit product ions that involve the fragmentation of D ring, as well as additional exclusive product ions. The introduction of an oxygenated function at E ring enhances the progress of fragmentation reactions through the β-keto ester group, developing also exclusive product ions for 13(2)-hydroxy derivatives and for 15(1)-hydroxy-lactone ones. Consequently, while MS(2)-based reactions of phytylated chlorophyll derivatives point to fragmentations at the phytyl and propionic chains, dephytylated chlorophyll derivatives behave different as the absence of phytyl makes β-keto ester group and E ring more prone to fragmentation. Proposals of the key reaction mechanisms underlying the origin of new product ions have been made. Copyright © 2015 Elsevier B.V. All rights reserved.

  16. Frechet derivatives for shallow water ocean acoustic inverse problems

    NASA Astrophysics Data System (ADS)

    Odom, Robert I.

    2003-04-01

    For any inverse problem, finding a model fitting the data is only half the problem. Most inverse problems of interest in ocean acoustics yield nonunique model solutions, and involve inevitable trade-offs between model and data resolution and variance. Problems of uniqueness and resolution and variance trade-offs can be addressed by examining the Frechet derivatives of the model-data functional with respect to the model variables. Tarantola [Inverse Problem Theory (Elsevier, Amsterdam, 1987), p. 613] published analytical formulas for the basic derivatives, e.g., derivatives of pressure with respect to elastic moduli and density. Other derivatives of interest, such as the derivative of transmission loss with respect to attenuation, can be easily constructed using the chain rule. For a range independent medium the analytical formulas involve only the Green's function and the vertical derivative of the Green's function for the medium. A crucial advantage of the analytical formulas for the Frechet derivatives over numerical differencing is that they can be computed with a single pass of any program which supplies the Green's function. Various derivatives of interest in shallow water ocean acoustics are presented and illustrated by an application to the sensitivity of measured pressure to shallow water sediment properties. [Work supported by ONR.

  17. New Metabolites and Bioactive Chlorinated Benzophenone Derivatives Produced by a Marine-Derived Fungus Pestalotiopsis heterocornis.

    PubMed

    Lei, Hui; Lin, Xiuping; Han, Li; Ma, Jian; Ma, Qingjuan; Zhong, Jialiang; Liu, Yonghong; Sun, Tiemin; Wang, Jinhui; Huang, Xueshi

    2017-03-13

    Four new compounds, including two isocoumarins, pestaloisocoumarins A and B ( 1 , 2 ), one sesquiterpenoid degradation, isopolisin B ( 4 ), and one furan derivative, pestalotiol A ( 5 ), together with one known isocoumarin, gamahorin ( 3 ), and three chlorinated benzophenone derivatives, pestalachloride B ( 6 ), pestalachloride E ( 7 ) and a mixture of pestalalactone atropisomers ( 8a/8b ), were isolated from a culture of the fungus Pestalotiopsis heterocornis associated with sponge Phakellia fusca . These new chemical structures were established using NMR and MS spectroscopic data, as well as single-crystal X-ray crystallographic analysis and CD Cotton effects. All of the isolated compounds were evaluated for their antimicrobial and cytotoxic activities. Isocoumarins 1 - 3 , showed antibacterial activities against Gram-positive bacteria Staphylococcus aureus and Bacillus subtilis with MIC values ranging from 25 to 100 μg/mL and weak antifungal activities. Chlorinated benzophenone derivatives 6 - 8 exhibited antibacterial activities against S. aureus and B. subtilis with MIC values ranging from 3.0 to 50 μg/mL and cytotoxicities against four human cancer cell lines with IC 50 values of 6.8-87.8 μM.

  18. Few Fractional Order Derivatives and Their Computations

    ERIC Educational Resources Information Center

    Bhatta, D. D.

    2007-01-01

    This work presents an introductory development of fractional order derivatives and their computations. Historical development of fractional calculus is discussed. This paper presents how to obtain computational results of fractional order derivatives for some elementary functions. Computational results are illustrated in tabular and graphical…

  19. Optimum sensitivity derivatives of objective functions in nonlinear programming

    NASA Technical Reports Server (NTRS)

    Barthelemy, J.-F. M.; Sobieszczanski-Sobieski, J.

    1983-01-01

    The feasibility of eliminating second derivatives from the input of optimum sensitivity analyses of optimization problems is demonstrated. This elimination restricts the sensitivity analysis to the first-order sensitivity derivatives of the objective function. It is also shown that when a complete first-order sensitivity analysis is performed, second-order sensitivity derivatives of the objective function are available at little additional cost. An expression is derived whose application to linear programming is presented.

  20. Human plasma platelet-derived exosomes: effects of aspirin.

    PubMed

    Goetzl, Edward J; Goetzl, Laura; Karliner, Joel S; Tang, Norina; Pulliam, Lynn

    2016-05-01

    Platelet-derived exosomes mediate platelet atherogenic interactions with endothelial cells and monocytes. A new method for isolation of plasma platelet-derived exosomes is described and used to examine effects of aging and aspirin on exosome cargo proteins. Exosome secretion by purified platelets in vitro did not increase after exposure to thrombin or collagen, as assessed by exosome counts and quantification of the CD81 exosome marker. Thrombin and collagen increased exosome content of α-granule chemokines CXCL4 and CXCL7 and cytoplasmic high-mobility group box 1 (HMGB1) protein, but not membrane platelet glycoprotein VI (GPVI), with dependence on extracellular calcium. Aspirin consumption significantly blocked thrombin- and collagen-induced increases in exosome cargo levels of chemokines and HMGB1, without altering total exosome secretion or GPVI cargo. Plasma platelet-derived exosomes, enriched by absorption with mouse antihuman CD42b [platelet glycoprotein Ib (GPIb)] mAb, had sizes and cargo protein contents similar to those of exosomes from purified platelets. The plasma platelet-derived exosome number is lower and its chemokine and HMGB1 levels higher after age 65 yr. Aspirin consumption significantly suppressed cargo protein levels of plasma platelet-derived exosomes without altering total levels of exosomes. Cargo proteins of human plasma platelet-derived exosomes may biomark platelet abnormalities and in vivo effects of drugs.- Goetzl, E. J., Goetzl, L., Karliner, J. S., Tang, N., Pulliam, L. Human plasma platelet-derived exosomes: effects of aspirin. © FASEB.

  1. Shikonin and its derivatives: a patent review.

    PubMed

    Wang, Rubing; Yin, Runting; Zhou, Wen; Xu, Defeng; Li, Shaoshun

    2012-09-01

    Shikonin and its derivatives are the main components of red pigment extracts from Lithospermum erythrorhizon, whose medicinal properties have been confirmed for a long history, and have aroused great interest as the hallmark molecules responsible for their significant biological activities, especially for their striking anticancer effects. Areas covered in this paper include a review of the total synthesis, biological effects and mechanisms of shikonin and its derivatives for their anticancer activities in the past decade, basing on literature and patents. The current state and problems are also discussed. At present, screening for anticancer shikonin derivatives is based on cellular level to find compounds with stronger cytotoxicity. Though several compounds have been discovered with striking cytotoxicity in vitro, however, no selectivity was observed and undoubtedly, the further outcomes have been disappointing because of their great damage to normal cells. Meanwhile, the presumed mechanisms of action are also established in terms of their cytotoxicity. From a pharmacological point of view, most of the shikonin derivatives are at an early stage of their development, and thus it is difficult to determine the exact effectiveness in cancer treatment. With research in this field going deeper, it can be expected that, despite the difficulties, shikonin derivatives as potential anticancer agents will soon follow.

  2. First Total Syntheses and Antimicrobial Evaluation of Penicimonoterpene, a Marine-Derived Monoterpenoid, and Its Various Derivatives

    PubMed Central

    Zhao, Jian-Chun; Li, Xiao-Ming; Gloer, James B.; Wang, Bin-Gui

    2014-01-01

    The first total synthesis of marine-derived penicimonoterpene (±)-1 has been achieved in four steps from 6-methylhept-5-en-2-one using a Reformatsky reaction as the key step to construct the basic carbon skeleton. A total of 24 new derivatives of 1 have also been designed and synthesized. Their structures were characterized by analysis of their 1H NMR, 13C NMR and HRESIMS data. Some of them showed significant antibacterial activity against Aeromonas hydrophila, Escherichia coli, Micrococcus luteus, Staphylococcus aureus, Vibrio anguillarum, V. harveyi and/or V. parahaemolyticus, and some showed activity against plant-pathogenic fungi (Alternaria brassicae, Colletotrichum gloeosporioides and/or Fusarium graminearum). Some of the derivatives exhibited antimicrobial MIC values ranging from 0.25 to 4 μg/mL, which were stronger than those of the positive control. Notably, Compounds 3b and 10 showed extremely high selectively against plant-pathogenic fungus F. graminearum (MIC 0.25 μg/mL) and pathogenic bacteria E. coli (MIC 1 μg/mL), implying their potential as antimicrobial agents. SAR analysis of 1 and its derivatives indicated that modification of the carbon-carbon double bond at C-6/7, of groups on the allylic methylene unit and of the carbonyl group at C-1, effectively enhanced the antimicrobial activity. PMID:24897384

  3. Comparison Of Quantitative Precipitation Estimates Derived From Rain Gauge And Radar Derived Algorithms For Operational Flash Flood Support.

    NASA Astrophysics Data System (ADS)

    Streubel, D. P.; Kodama, K.

    2014-12-01

    To provide continuous flash flood situational awareness and to better differentiate severity of ongoing individual precipitation events, the National Weather Service Research Distributed Hydrologic Model (RDHM) is being implemented over Hawaii and Alaska. In the implementation process of RDHM, three gridded precipitation analyses are used as forcing. The first analysis is a radar only precipitation estimate derived from WSR-88D digital hybrid reflectivity, a Z-R relationship and aggregated into an hourly ¼ HRAP grid. The second analysis is derived from a rain gauge network and interpolated into an hourly ¼ HRAP grid using PRISM climatology. The third analysis is derived from a rain gauge network where rain gauges are assigned static pre-determined weights to derive a uniform mean areal precipitation that is applied over a catchment on a ¼ HRAP grid. To assess the effect of different QPE analyses on the accuracy of RDHM simulations and to potentially identify a preferred analysis for operational use, each QPE was used to force RDHM to simulate stream flow for 20 USGS peak flow events. An evaluation of the RDHM simulations was focused on peak flow magnitude, peak flow timing, and event volume accuracy to be most relevant for operational use. Results showed RDHM simulations based on the observed rain gauge amounts were more accurate in simulating peak flow magnitude and event volume relative to the radar derived analysis. However this result was not consistent for all 20 events nor was it consistent for a few of the rainfall events where an annual peak flow was recorded at more than one USGS gage. Implications of this indicate that a more robust QPE forcing with the inclusion of uncertainty derived from the three analyses may provide a better input for simulating extreme peak flow events.

  4. The derivative-free Fourier shell identity for photoacoustics.

    PubMed

    Baddour, Natalie

    2016-01-01

    In X-ray tomography, the Fourier slice theorem provides a relationship between the Fourier components of the object being imaged and the measured projection data. The Fourier slice theorem is the basis for X-ray Fourier-based tomographic inversion techniques. A similar relationship, referred to as the 'Fourier shell identity' has been previously derived for photoacoustic applications. However, this identity relates the pressure wavefield data function and its normal derivative measured on an arbitrary enclosing aperture to the three-dimensional Fourier transform of the enclosed object evaluated on a sphere. Since the normal derivative of pressure is not normally measured, the applicability of the formulation is limited in this form. In this paper, alternative derivations of the Fourier shell identity in 1D, 2D polar and 3D spherical polar coordinates are presented. The presented formulations do not require the normal derivative of pressure, thereby lending the formulas directly adaptable for Fourier based absorber reconstructions.

  5. Robust Derivation of Risk Reduction Strategies

    NASA Technical Reports Server (NTRS)

    Richardson, Julian; Port, Daniel; Feather, Martin

    2007-01-01

    Effective risk reduction strategies can be derived mechanically given sufficient characterization of the risks present in the system and the effectiveness of available risk reduction techniques. In this paper, we address an important question: can we reliably expect mechanically derived risk reduction strategies to be better than fixed or hand-selected risk reduction strategies, given that the quantitative assessment of risks and risk reduction techniques upon which mechanical derivation is based is difficult and likely to be inaccurate? We consider this question relative to two methods for deriving effective risk reduction strategies: the strategic method defined by Kazman, Port et al [Port et al, 2005], and the Defect Detection and Prevention (DDP) tool [Feather & Cornford, 2003]. We performed a number of sensitivity experiments to evaluate how inaccurate knowledge of risk and risk reduction techniques affect the performance of the strategies computed by the Strategic Method compared to a variety of alternative strategies. The experimental results indicate that strategies computed by the Strategic Method were significantly more effective than the alternative risk reduction strategies, even when knowledge of risk and risk reduction techniques was very inaccurate. The robustness of the Strategic Method suggests that its use should be considered in a wide range of projects.

  6. High speed point derivative microseismic detector

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

    Uhl, J.E.; Warpinski, N.R.; Whetten, E.B.

    A high speed microseismic event detector constructed in accordance with the present invention uses a point derivative comb to quickly and accurately detect microseismic events. Compressional and shear waves impinging upon microseismic receiver stations disposed to collect waves are converted into digital data and analyzed using a point derivative comb including assurance of quiet periods prior to declaration of microseismic events. If a sufficient number of quiet periods have passed, the square of a two point derivative of the incoming digital signal is compared to a trip level threshold exceeding the determined noise level to declare a valid trial event.more » The squaring of the derivative emphasizes the differences between noise and signal, and the valid event is preferably declared when the trip threshold has been exceeded over a temporal comb width to realize a comb over a given time period. Once a trial event has been declared, the event is verified through a spatial comb, which applies the temporal event comb to additional stations. The detector according to the present invention quickly and accurately detects initial compressional waves indicative of a microseismic event which typically exceed the ambient cultural noise level by a small amount, and distinguishes the waves from subsequent larger amplitude shear waves. 9 figs.« less

  7. High speed point derivative microseismic detector

    DOEpatents

    Uhl, J.E.; Warpinski, N.R.; Whetten, E.B.

    1998-06-30

    A high speed microseismic event detector constructed in accordance with the present invention uses a point derivative comb to quickly and accurately detect microseismic events. Compressional and shear waves impinging upon microseismic receiver stations disposed to collect waves are converted into digital data and analyzed using a point derivative comb including assurance of quiet periods prior to declaration of microseismic events. If a sufficient number of quiet periods have passed, the square of a two point derivative of the incoming digital signal is compared to a trip level threshold exceeding the determined noise level to declare a valid trial event. The squaring of the derivative emphasizes the differences between noise and signal, and the valid event is preferably declared when the trip threshold has been exceeded over a temporal comb width to realize a comb over a given time period. Once a trial event has been declared, the event is verified through a spatial comb, which applies the temporal event comb to additional stations. The detector according to the present invention quickly and accurately detects initial compressional waves indicative of a microseismic event which typically exceed the ambient cultural noise level by a small amount, and distinguishes the waves from subsequent larger amplitude shear waves. 9 figs.

  8. High speed point derivative microseismic detector

    DOEpatents

    Uhl, James Eugene; Warpinski, Norman Raymond; Whetten, Ernest Blayne

    1998-01-01

    A high speed microseismic event detector constructed in accordance with the present invention uses a point derivative comb to quickly and accurately detect microseismic events. Compressional and shear waves impinging upon microseismic receiver stations disposed to collect waves are converted into digital data and analyzed using a point derivative comb including assurance of quiet periods prior to declaration of microseismic events. If a sufficient number of quiet periods have passed, the square of a two point derivative of the incoming digital signal is compared to a trip level threshold exceeding the determined noise level to declare a valid trial event. The squaring of the derivative emphasizes the differences between noise and signal, and the valid event is preferably declared when the trip threshold has been exceeded over a temporal comb width to realize a comb over a given time period. Once a trial event has been declared, the event is verified through a spatial comb, which applies the temporal event comb to additional stations. The detector according to the present invention quickly and accurately detects initial compressional waves indicative of a microseismic event which typically exceed the ambient cultural noise level by a small amount, and distinguishes the waves from subsequent larger amplitude shear waves.

  9. 76 FR 51878 - Modifications of Certain Derivative Contracts; Correction

    Federal Register 2010, 2011, 2012, 2013, 2014

    2011-08-19

    ... Modifications of Certain Derivative Contracts; Correction AGENCY: Internal Revenue Service (IRS), Treasury... temporary regulations (TD 9538) that address when a transfer or assignment of certain derivative contracts... read as follows: Sec. 1.1001-4T Modifications of certain derivative contracts (temporary). * * * * * (a...

  10. 21 CFR 173.135 - Catalase derived from Micrococcus lysodeikticus.

    Code of Federal Regulations, 2011 CFR

    2011-04-01

    ... 21 Food and Drugs 3 2011-04-01 2011-04-01 false Catalase derived from Micrococcus lysodeikticus... FOOD FOR HUMAN CONSUMPTION Enzyme Preparations and Microorganisms § 173.135 Catalase derived from Micrococcus lysodeikticus. Bacterial catalase derived from Micrococcus lysodeikticus by a pure culture...

  11. 14 CFR 1203.501 - Applying derivative classification markings.

    Code of Federal Regulations, 2011 CFR

    2011-01-01

    ... INFORMATION SECURITY PROGRAM Derivative Classification § 1203.501 Applying derivative classification markings... classification decisions: (b) Verify the information's current level of classification so far as practicable...

  12. 14 CFR 1203.501 - Applying derivative classification markings.

    Code of Federal Regulations, 2010 CFR

    2010-01-01

    ... INFORMATION SECURITY PROGRAM Derivative Classification § 1203.501 Applying derivative classification markings... classification decisions: (b) Verify the information's current level of classification so far as practicable...

  13. Derivation of Formulations 1 and 1A of Farassat

    NASA Technical Reports Server (NTRS)

    Farassat, F.

    2007-01-01

    Formulations 1 and 1A are the solutions of the Ffowcs Williams-Hawkings (FW-H) equation with surface sources only when the surface moves at subsonic speed. Both formulations have been successfully used for helicopter rotor and propeller noise prediction for many years although we now recommend using Formulation 1A for this purpose. Formulation 1 has an observer time derivative that is taken numerically, and thus, increasing execution time on a computer and reducing the accuracy of the results. After some discussion of the Green's function of the wave equation, we derive Formulation 1 which is the basis of deriving Formulation 1A. We will then show how to take this observer time derivative analytically to get Formulation 1A. We give here the most detailed derivation of these formulations. Once you see the whole derivation, you will ask yourself why you did not do it yourself!

  14. Modelling of Piezothermoelastic Beam with Fractional Order Derivative

    NASA Astrophysics Data System (ADS)

    Kumar, Rajneesh; Sharma, Poonam

    2016-04-01

    This paper deals with the study of transverse vibrations in piezothermoelastic beam resonators with fractional order derivative. The fractional order theory of thermoelasticity developed by Sherief et al. [1] has been used to study the problem. The expressions for frequency shift and damping factor are derived for a thermo micro-electromechanical (MEM) and thermo nano-electromechanical (NEM) beam resonators clamped on one side and free on another. The effect of fractional order derivative on the derived expressions is observed analytically and shown graphically in the case of Lead Zirconate Titanate (PZT)-5A material. For α = 1, our results agree with those that are obtained by Grover and Sharma [20] and other particular cases of interest are also discussed.

  15. Pleuromutilin and its derivatives-the lead compounds for novel antibiotics.

    PubMed

    Tang, Y-Z; Liu, Y-H; Chen, J-X

    2012-01-01

    Due to the rapid onset of resistance to most antibacterial drugs, research efforts are focusing on new classes of antibacterials with different mechanisms of action from clinically used antibacterials. Pleuromutilin derivatives have received more and more scientific attention for their unique mechanism of action. Two pleuromutilin derivatives, tiamulin and valnemulin have been successfully developed as antibiotics for veterinary use. Retapamulin, another pleuromutilin derivative has been approved for use in humans in April 2007 by Food and Drug Administration (FDA). It has been shown that there is rarely cross-resistance between pleuromutilin derivatives and other antimicrobial agents, and the development of resistance bacterial is still low. This review will demonstrate mechanism of action of pleuromutilin derivatives and reveal the structure-activity relationship (SAR) of pleuromutilin derivatives. Additionally, the pleuromutilin antibacterial derivative agents in the market, such as tiamulin, valnemulin and retapamulin, will be discussed. It is proposed that new antibacterial agents might be developed from pleuromutilin derivatives in the future.

  16. Increased circulating leukocyte-derived microparticles in ischemic cerebrovascular disease.

    PubMed

    He, Zhangping; Tang, Yanyan; Qin, Chao

    2017-06-01

    Circulating leukocyte-derived microparticles act as proinflammatory mediators that reflect vascular inflammation. In this study, we examined the hypothesis that the quantity of leukocyte-derived microparticles is increased in patients with ischemic cerebrovascular diseases, and investigated utility of various phenotypes of leukocyte-derived microparticles as specific biomarkers of vascular inflammation injury. Additionally we focused on identifying leukocyte-derived microparticles that may be correlated with stroke severity in acute ischemic stroke patients. The plasma concentration of leukocyte-derived microparticles obtained by a series of centrifugations of 76 consecutive patients with ischemic cerebrovascular diseases and 70 age-, sex-, and race-matched healthy controls were determined by flow cytometry. Significantly elevated numbers of leukocyte (CD45+), monocyte (CD14+), lymphocyte (CD4+), granulocyte (CD15+) derived microparticles were found in the plasma samples of patients ischemic cerebrovascular diseases, compared to healthy controls (p<0.05). Furthermore, the plasma levels of CD14+ microparticles were significantly correlated with stroke severity (r=0.355, p=0.019), cerebral vascular stenosis severity (r=0.255, p=0.025) and stroke subtype (r=0.242, p=0.036). No association with stroke was observed for other leukocyte-derived phenotypes. These results demonstrate that circulating leukocyte-derived microparticles amounts are increased in patients with ischemic cerebrovascular diseases, compared with healthy controls. As proinflammatory mediators, leukocyte-derived microparticles may contribute to vascular inflammatory and the inflammatory process in acute ischemic stroke. Levels of CD14+ microparticles may be a promising biomarker of ischemic severity and outcome of stroke in the clinic. Copyright © 2017 Elsevier Ltd. All rights reserved.

  17. 40 CFR 721.5925 - Bis heterocyclic phenylene derivative (generic).

    Code of Federal Regulations, 2010 CFR

    2010-07-01

    ... 40 Protection of Environment 30 2010-07-01 2010-07-01 false Bis heterocyclic phenylene derivative... Specific Chemical Substances § 721.5925 Bis heterocyclic phenylene derivative (generic). (a) Chemical... as bis heterocyclic phenylene derivative (PMN P-01-0432) is subject to reporting under this section...

  18. 40 CFR 721.5925 - Bis heterocyclic phenylene derivative (generic).

    Code of Federal Regulations, 2011 CFR

    2011-07-01

    ... 40 Protection of Environment 31 2011-07-01 2011-07-01 false Bis heterocyclic phenylene derivative... Specific Chemical Substances § 721.5925 Bis heterocyclic phenylene derivative (generic). (a) Chemical... as bis heterocyclic phenylene derivative (PMN P-01-0432) is subject to reporting under this section...

  19. 76 FR 51878 - Modifications of Certain Derivative Contracts; Correction

    Federal Register 2010, 2011, 2012, 2013, 2014

    2011-08-19

    ... Modifications of Certain Derivative Contracts; Correction AGENCY: Internal Revenue Service (IRS), Treasury... derivative contracts does not result in an exchange to the nonassigning counterparty for purposes of Sec. 1... derivative contract to another dealer in securities or'' is corrected to read ``or a clearinghouse transfers...

  20. Development and application of an excitation ratiometric optical pH sensor for bioprocess monitoring.

    PubMed

    Badugu, Ramachandram; Kostov, Yordan; Rao, Govind; Tolosa, Leah

    2008-01-01

    The development of a fluorescent excitation ratiometric pH sensor (AHQ-PEG) using a novel allylhydroxyquinolinium (AHQ) derivative copolymerized with polyethylene glycol dimethacrylate (PEG) is described. The AHQ-PEG sensor film is shown to be suitable for real-time, noninvasive, continuous, online pH monitoring of bioprocesses. Optical ratiometric measurements are generally more reliable, robust, inexpensive, and insensitive to experimental errors such as fluctuations in the source intensity and fluorophore photobleaching. The sensor AHQ-PEG in deionized water was shown to exhibit two excitation maxima at 375 and 425 nm with a single emission peak at 520 nm. Excitation spectra of AHQ-PEG show a decrease in emission at the 360 nm excitation and an increase at the 420 nm excitation with increasing pH. Accordingly, the ratio of emission at 420:360 nm excitation showed a maximum change between pH 5 and 8 with an apparent pK(a) of 6.40. The low pK(a) value is suitable for monitoring the fermentation of most industrially important microorganisms. Additionally, the AHQ-PEG sensor was shown to have minimal sensitivity to ionic strength and temperature. Because AHQ is covalently attached to PEG, the film shows no probe leaching and is sterilizable by steam and alcohol. It shows rapid (approximately 2 min) and reversible response to pH over many cycles without any photobleaching. Subsequently, the AHQ-PEG sensor film was tested for its suitability in monitoring the pH of S. cereviseae (yeast) fermentation. The observed pH using AHQ-PEG film is in agreement with a conventional glass pH electrode. However, unlike the glass electrode, the present sensor is easily adaptable to noninvasive monitoring of sterilized, closed bioprocess environments without the awkward wire connections that electrodes require. In addition, the AHQ-PEG sensor is easily miniaturized to fit in microwell plates and microbioreactors for high-throughput cell culture applications.

  1. How predictive could alchemical derivatives be?

    PubMed

    Muñoz, Macarena; Cárdenas, Carlos

    2017-06-21

    The chemical space contains all possible compounds that can be imagined. Its size easily equals the number of fundamental particles in the observable universe. Rational design of compounds aims to find those sectors of the chemical space where compounds optimize a set of desired properties. Then, rational design demands tools to efficiently navigate the chemical space. Ab initio alchemical derivatives offer the possibility to navigate, without empiricism, the energy landscape through alchemical transformations. An alchemical transformation is any process, physical or fictitious, that connects to points in the chemical space. In this work, those transformations are constructed as a perturbative expansion of the energy with respect to perturbations in the stoichiometry. The response functions of that expansion are what is called alchemical derivatives. In this work we assess how effective alchemical derivatives are in predicting energy changes associated to changes in the composition. We do this by including in the expansion, for the first time, electrostatic, polarization and electron-transfer effects. The system we chose is one that challenges alchemical derivatives because none of these effects dominates its behavior. The transmutations studied here correspond to substitutional doping of Al 13 with up to four atoms of Si, Al 13-n Si n . Two types of transformations are considered, those in which the number of electrons remains constant and those in which the number of electrons also changes. It is found that contrary to what has been reported before, polarization cannot be neglected. If polarization is not included, alchemical derivatives fail to predict the change of energy and the relative energy between isomers. For isoelectronic substitution of four or more atoms, the perturbative approach collapses because the strength of the perturbation becomes too strong to guarantee convergence of the series. It is shown, however, that if only one atom is mutated at a

  2. 1,3-Oxazin-6-one Derivatives and Bohemamine-Type Pyrrolizidine Alkaloids from a Marine-Derived Streptomyces spinoverrucosus

    PubMed Central

    2015-01-01

    Two new 1,3-oxazin-6-one derivatives (1 and 2) and six new bohemamine-type pyrrolizidine alkaloids (3–8) were isolated from the marine-derived Streptomyces spinoverrucosus strain SNB-048. Their structures including the absolute configurations were fully elucidated on the basis of spectroscopic analysis, ECD spectra, quantum chemical calculations, and chemical methods. Compounds 1 and 2 possess a γ-lactam moiety and a 1,3-oxazin-6-one system. PMID:26489038

  3. Shoot-derived abscisic acid promotes root growth.

    PubMed

    McAdam, Scott A M; Brodribb, Timothy J; Ross, John J

    2016-03-01

    The phytohormone abscisic acid (ABA) plays a major role in regulating root growth. Most work to date has investigated the influence of root-sourced ABA on root growth during water stress. Here, we tested whether foliage-derived ABA could be transported to the roots, and whether this foliage-derived ABA had an influence on root growth under well-watered conditions. Using both application studies of deuterium-labelled ABA and reciprocal grafting between wild-type and ABA-biosynthetic mutant plants, we show that both ABA levels in the roots and root growth in representative angiosperms are controlled by ABA synthesized in the leaves rather than sourced from the roots. Foliage-derived ABA was found to promote root growth relative to shoot growth but to inhibit the development of lateral roots. Increased root auxin (IAA) levels in plants with ABA-deficient scions suggest that foliage-derived ABA inhibits root growth through the root growth-inhibitor IAA. These results highlight the physiological and morphological importance, beyond the control of stomata, of foliage-derived ABA. The use of foliar ABA as a signal for root growth has important implications for regulating root to shoot growth under normal conditions and suggests that leaf rather than root hydration is the main signal for regulating plant responses to moisture. © 2015 John Wiley & Sons Ltd.

  4. Multiscale analysis of the CMB temperature derivatives

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

    Marcos-Caballero, A.; Martínez-González, E.; Vielva, P., E-mail: marcos@ifca.unican.es, E-mail: martinez@ifca.unican.es, E-mail: vielva@ifca.unican.es

    2017-02-01

    We study the Planck CMB temperature at different scales through its derivatives up to second order, which allows one to characterize the local shape and isotropy of the field. The problem of having an incomplete sky in the calculation and statistical characterization of the derivatives is addressed in the paper. The analysis confirms the existence of a low variance in the CMB at large scales, which is also noticeable in the derivatives. Moreover, deviations from the standard model in the gradient, curvature and the eccentricity tensor are studied in terms of extreme values on the data. As it is expected,more » the Cold Spot is detected as one of the most prominent peaks in terms of curvature, but additionally, when the information of the temperature and its Laplacian are combined, another feature with similar probability at the scale of 10{sup o} is also observed. However, the p -value of these two deviations increase above the 6% when they are referred to the variance calculated from the theoretical fiducial model, indicating that these deviations can be associated to the low variance anomaly. Finally, an estimator of the directional anisotropy for spinorial quantities is introduced, which is applied to the spinors derived from the field derivatives. An anisotropic direction whose probability is <1% is detected in the eccentricity tensor.« less

  5. Treating autoimmune disorders with venom-derived peptides.

    PubMed

    Shen, Bingzheng; Cao, Zhijian; Li, Wenxin; Sabatier, Jean-Marc; Wu, Yingliang

    2017-09-01

    The effective treatment of autoimmune diseases remains a challenge. Voltage-gated potassium Kv1.3 channels, which are expressed in lymphocytes, are a new therapeutic target for treating autoimmune disease. Consequently, Kv1.3 channel-inhibiting venom-derived peptides are a prospective resource for new drug discovery and clinical application. Area covered: Preclinical and clinical studies have produced a wealth of information on Kv1.3 channel-inhibiting venom-derived peptides, especially from venomous scorpions and sea anemones. This review highlights the advances in screening and design of these peptides with diverse structures and potencies. It focuses on representative strategies for improving peptide selectivity and discusses the preclinical research on those venom-derived peptides as well as their clinical developmental status. Expert opinion: Encouraging results indicate that peptides isolated from the venom of venomous animals are a large resource for discovering immunomodulators that act on Kv1.3 channels. Since the structural diversity of venom-derived peptides determines the variety of their pharmacological activities, the design and optimization of venom-peptides for improved Kv1.3 channel-specificity has been advanced through some representative strategies, such as peptide chemical modification, amino acid residue truncation and binding interface modulation. These advances should further accelerate research, development and the future clinical application of venom-derived peptides selectively targeting Kv1.3 channels.

  6. 21 CFR 184.1521 - Monosodium phosphate derivatives of mono- and diglycerides.

    Code of Federal Regulations, 2011 CFR

    2011-04-01

    ... 21 Food and Drugs 3 2011-04-01 2011-04-01 false Monosodium phosphate derivatives of mono- and... derivatives of mono- and diglycerides. (a) Monosodium phophate derivatives of mono- and diglycerides are composed of glyceride derivatives formed by reacting mono- and diglycerides that are derived from edible...

  7. 18 CFR 367.2440 - Account 244, Derivative instrument liabilities.

    Code of Federal Regulations, 2012 CFR

    2012-04-01

    ... 18 Conservation of Power and Water Resources 1 2012-04-01 2012-04-01 false Account 244, Derivative instrument liabilities. 367.2440 Section 367.2440 Conservation of Power and Water Resources FEDERAL ENERGY..., Derivative instrument liabilities. This account must include the change in the fair value of all derivative...

  8. 18 CFR 367.2440 - Account 244, Derivative instrument liabilities.

    Code of Federal Regulations, 2010 CFR

    2010-04-01

    ... 18 Conservation of Power and Water Resources 1 2010-04-01 2010-04-01 false Account 244, Derivative instrument liabilities. 367.2440 Section 367.2440 Conservation of Power and Water Resources FEDERAL ENERGY..., Derivative instrument liabilities. This account must include the change in the fair value of all derivative...

  9. 40 CFR 721.6097 - Phosphoric acid derivative (generic name).

    Code of Federal Regulations, 2010 CFR

    2010-07-01

    ... 40 Protection of Environment 30 2010-07-01 2010-07-01 false Phosphoric acid derivative (generic... Specific Chemical Substances § 721.6097 Phosphoric acid derivative (generic name). (a) Chemical substance... phosphoric acid derivative (PMN P-95-284) is subject to reporting under this section for the significant new...

  10. 40 CFR 721.6097 - Phosphoric acid derivative (generic name).

    Code of Federal Regulations, 2011 CFR

    2011-07-01

    ... 40 Protection of Environment 31 2011-07-01 2011-07-01 false Phosphoric acid derivative (generic... Specific Chemical Substances § 721.6097 Phosphoric acid derivative (generic name). (a) Chemical substance... phosphoric acid derivative (PMN P-95-284) is subject to reporting under this section for the significant new...

  11. Antioxidant and antimicrobial activities of cinnamic acid derivatives.

    PubMed

    Sova, M

    2012-07-01

    Cinnamic acid is an organic acid occurring naturally in plants that has low toxicity and a broad spectrum of biological activities. In the search for novel pharmacologically active compounds, cinnamic acid derivatives are important and promising compounds with high potential for development into drugs. Many cinnamic acid derivatives, especially those with the phenolic hydroxyl group, are well-known antioxidants and are supposed to have several health benefits due to their strong free radical scavenging properties. It is also well known that cinnamic acid has antimicrobial activity. Cinnamic acid derivatives, both isolated from plant material and synthesized, have been reported to have antibacterial, antiviral and antifungal properties. Acids, esters, amides, hydrazides and related derivatives of cinnamic acid with such activities are here reviewed.

  12. On the role of second number-conserving functional derivatives

    NASA Astrophysics Data System (ADS)

    Gál, Tamás

    2006-06-01

    It is found that number-conserving second derivatives, of functional differentiation constrained to the domain of functional variables ρ(x) of a given norm ∫ρ(x)dx, are not obtained via two successive number-conserving differentiations, contrary to the case of unrestricted second derivatives. Investigating the role of second number-conserving derivatives, with the density-functional formulation of time-dependent quantum mechanics in focus, it is shown how number-conserving differentiation handles the dual nature of the Kohn Sham potential arising in the practical use of the theory. On the other hand, it is pointed out that number-conserving derivatives cannot resolve the causality paradox connected with the second derivative of the exchange-correlation part of the action density functional.

  13. Evaluation of antituberculosis activity and DFT study on dipyrromethane-derived hydrazone derivatives

    NASA Astrophysics Data System (ADS)

    Rawat, Poonam; Singh, R. N.; Niranjan, Priydarshni; Ranjan, Alok; Holguín, Norma Rosario Flores

    2017-12-01

    This paper evaluates the anti-tubercular activity of dipyrromethane-derived hydrazones derivatives (3a-d) against strain of Mycobacterium tuberculosis H37Rv. The newly synthesized compounds have been obtained in good yield based on the condensation of aromatic aldehyde derivatives with pyrrole hydrazone in presence of catalyst and well characterized with spectroscopic methods (1H, 13C NMR, Mass spectrometry) and elemental analysis. The singlet observed in the experimental 1H and 13C NMR spectra in the range of 5.3-5.7 ppm and 30-33.86 ppm, respectively, indicating that two pyrrole units are joined at meso position. The electronic transitions observed in the experimental spectra are n→π* and π →π* in nature. Experimental and theoretical findings corroborate well with each other. The substitution of acceptor group (-NO2) at ortho- and meta-positions of benzene ring, present at meso-position of dipyrromethane is responsible for variation in β0 values. The calculated NLO of (3a-d) are much greater than those of p-nitroaniline (PNA). The solvent induced effects on the non-linear optical properties were studied and found to enhance NLO properties of the molecules as dielectric constants of the solvents increases. On the basis of results it is anticipated that these dipyrromethanes will be useful for both antimicrobial and non-linear optical (NLO) applications. With the help of Microplate Alamar Blue assay (MABA) method all (3a-d) compounds were screened for their anti-tubercular activity and found that 3b and 3d have higher inhibitory activity against strain of M. tuberculosis H37Rv.

  14. Cloud vertical profiles derived from CALIPSO and CloudSat and a comparison with MODIS derived clouds

    NASA Astrophysics Data System (ADS)

    Kato, S.; Sun-Mack, S.; Miller, W. F.; Rose, F. G.; Minnis, P.; Wielicki, B. A.; Winker, D. M.; Stephens, G. L.; Charlock, T. P.; Collins, W. D.; Loeb, N. G.; Stackhouse, P. W.; Xu, K.

    2008-05-01

    CALIPSO and CloudSat from the a-train provide detailed information of vertical distribution of clouds and aerosols. The vertical distribution of cloud occurrence is derived from one month of CALIPSO and CloudSat data as a part of the effort of merging CALIPSO, CloudSat and MODIS with CERES data. This newly derived cloud profile is compared with the distribution of cloud top height derived from MODIS on Aqua from cloud algorithms used in the CERES project. The cloud base from MODIS is also estimated using an empirical formula based on the cloud top height and optical thickness, which is used in CERES processes. While MODIS detects mid and low level clouds over the Arctic in April fairly well when they are the topmost cloud layer, it underestimates high- level clouds. In addition, because the CERES-MODIS cloud algorithm is not able to detect multi-layer clouds and the empirical formula significantly underestimates the depth of high clouds, the occurrence of mid and low-level clouds is underestimated. This comparison does not consider sensitivity difference to thin clouds but we will impose an optical thickness threshold to CALIPSO derived clouds for a further comparison. The effect of such differences in the cloud profile to flux computations will also be discussed. In addition, the effect of cloud cover to the top-of-atmosphere flux over the Arctic using CERES SSF and FLASHFLUX products will be discussed.

  15. A simple derivation of Lorentz self-force

    NASA Astrophysics Data System (ADS)

    Haque, Asrarul

    2014-09-01

    We derive the Lorentz self-force for a charged particle in arbitrary non-relativistic motion by averaging the retarded fields. The derivation is simple and at the same time pedagogically accessible. We obtain the radiation reaction for a charged particle moving in a circle. We pin down the underlying concept of mass renormalization.

  16. Characterizing Thematized Derivative Schema by the Underlying Emergent Structures

    ERIC Educational Resources Information Center

    Garcia, Mercedes; Llinares, Salvador; Sanchez-Matamoros, Gloria

    2011-01-01

    This paper reports on different underlying structures of the derivative schema of three undergraduate students that were considered to be at the trans level of development of the derivative schema (action-process-object-schema). The derivative schema is characterized in terms of the students' ability to explicitly transfer the relationship between…

  17. 18 CFR 367.1750 - Account 175, Derivative instrument assets.

    Code of Federal Regulations, 2011 CFR

    2011-04-01

    ... 18 Conservation of Power and Water Resources 1 2011-04-01 2011-04-01 false Account 175, Derivative... GAS ACT Balance Sheet Chart of Accounts Current and Accrued Assets § 367.1750 Account 175, Derivative instrument assets. This account must include the amounts paid for derivative instruments, and the change in...

  18. New Metabolites and Bioactive Chlorinated Benzophenone Derivatives Produced by a Marine-Derived Fungus Pestalotiopsis heterocornis

    PubMed Central

    Lei, Hui; Lin, Xiuping; Han, Li; Ma, Jian; Ma, Qingjuan; Zhong, Jialiang; Liu, Yonghong; Sun, Tiemin; Wang, Jinhui; Huang, Xueshi

    2017-01-01

    Four new compounds, including two isocoumarins, pestaloisocoumarins A and B (1, 2), one sesquiterpenoid degradation, isopolisin B (4), and one furan derivative, pestalotiol A (5), together with one known isocoumarin, gamahorin (3), and three chlorinated benzophenone derivatives, pestalachloride B (6), pestalachloride E (7) and a mixture of pestalalactone atropisomers (8a/8b), were isolated from a culture of the fungus Pestalotiopsis heterocornis associated with sponge Phakellia fusca. These new chemical structures were established using NMR and MS spectroscopic data, as well as single-crystal X-ray crystallographic analysis and CD Cotton effects. All of the isolated compounds were evaluated for their antimicrobial and cytotoxic activities. Isocoumarins 1–3, showed antibacterial activities against Gram-positive bacteria Staphylococcus aureus and Bacillus subtilis with MIC values ranging from 25 to 100 μg/mL and weak antifungal activities. Chlorinated benzophenone derivatives 6–8 exhibited antibacterial activities against S. aureus and B. subtilis with MIC values ranging from 3.0 to 50 μg/mL and cytotoxicities against four human cancer cell lines with IC50 values of 6.8–87.8 μM. PMID:28335391

  19. Effect of molecular side groups and local nanoenvironment on photodegradation and its reversibility

    NASA Astrophysics Data System (ADS)

    Quist, Nicole; Li, Mark; Tollefsen, Ryan; Haley, Michael; Anthony, John; Ostroverkhova, Oksana

    2018-02-01

    Degradation of organic semiconductors in the presence of oxygen is one of the bottlenecks preventing their wide-spread use in optoelectronic devices. The first step towards such degradation in functionalized pentacene (Pn) derivatives is formation of endoperoxide (EPO), which can either revert back to the parent molecule or proceed to molecule decomposition. We present the study of reversibility of EPO formation through probing the photophysical properties of functionalized fluorinated pentacene (Pn-R-F8) derivatives. Experiments are done in solutions and in films both at the single molecule level and in the bulk. In solutions, degradation of optical absorption and its partial recovery after thermolysis were quantified for various derivatives depending on the solvent. At the single molecule level, low concentrations of each type of molecules were imaged in a variety of polymer matrices at 633 nm excitation at room temperature in air using wide-field fluorescence microscopy. Fluorescence time trajectories were collected and statistically analyzed to quantify blinking due to reversible EPO formation depending on the host matrix. To understand the physical changes of the molecular system, a Monte Carlo method was used to create a multi-level simulation, which enabled us to relate the change in the molecular transition rates to the experimentally measured parameters. At the bulk level, photoluminescence decay due to photobleaching and recovery due to EPO reconversion were measured for the same derivatives incorporated into various matrices. These studies provide insight into the synergistic effect of the local nanoenvironment and molecular side groups on the oxygen-related degradation and subsequent recovery which is important for development of organic electronic devices.

  20. Plasma-derived microparticles in polycythaemia vera.

    PubMed

    Ahadon, M; Abdul Aziz, S; Wong, C L; Leong, C F

    2018-04-01

    Microparticles are membrane bound vesicles, measuring less than 1.0 um, which are released during cellular activation or during apoptosis. Studies have shown that these circulating microparticles play a role in coagulation, cell signaling and cellular interactions. Increased levels of circulating microparticles have been observed in a number of conditions where there is vascular dysfunction, thrombosis and inflammation. The objective of this study was to determine the various plasma-derived microparticles in patients with polycythaemia vera (PV) in Universiti Kebangsaan Malaysia Medical Centre and to compare them with normal control. A total of 15 patients with PV and 15 healthy volunteers were included in this cross-sectional descriptive study. Plasma samples from both patients and healthy volunteers were prepared and further processed for isolation of microparticles. Flow cytometry analyses were then carried out in all samples to determine the cellular origin of the microparticles. Full blood count parameters for both groups were also collected. Data collected were analyzed using SPSS version 12.0. Patients with PV had a significantly higher percentage of platelet derived microparticles compared to healthy controls (P <0.05). The control group had a higher level of endothelial derived microparticles but the differences were not statistically significant (P > 0.05). The median percentage of positive events for platelet derived microparticles was higher in patients with PV compared to normal healthy controls.

  1. Understanding the Derivative through the Calculus Triangle

    ERIC Educational Resources Information Center

    Weber, Eric; Tallman, Michael; Byerley, Cameron; Thompson, Patrick W.

    2012-01-01

    Typical treatments of the derivative do not clearly convey the idea that the derivative function represents the original function's rate of change. Revealing the relationship between a function and its rate-of-change function for static values of "x" does not facilitate productive ways of thinking about generating the rate-of-change function or…

  2. Derive Workshop Matrix Algebra and Linear Algebra.

    ERIC Educational Resources Information Center

    Townsley Kulich, Lisa; Victor, Barbara

    This document presents the course content for a workshop that integrates the use of the computer algebra system Derive with topics in matrix and linear algebra. The first section is a guide to using Derive that provides information on how to write algebraic expressions, make graphs, save files, edit, define functions, differentiate expressions,…

  3. Space-Derived Transparency: Players, Policies, Implications, and Synergies

    NASA Astrophysics Data System (ADS)

    Kinnan, C. J.

    2001-06-01

    Space-derived transparency will become a common means of monitoring, preventing, and mitigating crises, verifying compliance with treaties and law, and enabling confidence and security building measures. Democratization and globalization, the proliferation of information technologies, the availability of commercial space high-resolution imagery, and the growing influence of NGOs invite this question: What is (space-derived) transparency and what effect does it have on US security policy? Three camps have emerged in the debate -Horaeists who seek to build a transnational society through complete transparency; Preservationists, mostly military, who fear the threat to national security, want to deny most space-derived information to non-traditional/non-state actors; and Synergists who seek to capitalize on the best of both camps. There is evidence suggesting that space-derived transparency is an inevitable trend and will resist even the best means of preservationist control. Space-derived transparency may change the dynamic of the security environment by introducing new players into the policy fomentation and implementation process. These players, if not properly schooled in imagery analysis or the potential effects of their use of misinterpreted space-derived imagery, could force policy makers to make fast, ill-considered decisions in order to respond to incidents. In some cases this fast response will defuse potential crises and in other situations these rushed decisions might result in policies without considering the potential consequences, which could turn incidents into crises. Space-derived transparency is a step forward into the future for each camp . . . the challenge for the United States lies in forging synergies in an increasingly transparent world while maintaining the balance between openness and security.

  4. A Generic Authentication LoA Derivation Model

    NASA Astrophysics Data System (ADS)

    Yao, Li; Zhang, Ning

    One way of achieving a more fine-grained access control is to link an authentication level of assurance (LoA) derived from a requester’s authentication instance to the authorisation decision made to the requester. To realise this vision, there is a need for designing a LoA derivation model that supports the use and quantification of multiple LoA-effecting attributes, and analyse their composite effect on a given authentication instance. This paper reports the design of such a model, namely a generic LoA derivation model (GEA- LoADM). GEA-LoADM takes into account of multiple authentication attributes along with their relationships, abstracts the composite effect by the multiple attributes into a generic value, authentication LoA, and provides algorithms for the run-time derivation of LoA. The algorithms are tailored to reflect the relationships among the attributes involved in an authentication instance. The model has a number of valuable properties, including flexibility and extensibility; it can be applied to different application contexts and support easy addition of new attributes and removal of obsolete ones.

  5. Marathon running increases circulating endothelial- and thrombocyte-derived microparticles.

    PubMed

    Schwarz, Viktoria; Düsing, Philip; Liman, Thomas; Werner, Christian; Herm, Juliane; Bachelier, Katrin; Krüll, Matthias; Brechtel, Lars; Jungehulsing, Gerhard J; Haverkamp, Wilhelm; Böhm, Michael; Endres, Matthias; Haeusler, Karl Georg; Laufs, Ulrich

    2018-02-01

    Background Acute vascular effects of high intensity physical activity are incompletely characterized. Circulating microparticles are cellular markers for vascular activation and damage. Methods Microparticles were analysed in 99 marathon runners (49 ± 6 years, 22% female) of the prospective Berlin Beat of Running study. Blood samples were taken within three days before, immediately after and within two days after the marathon run. Endothelial-derived microparticles were labelled with CD144, CD31 and CD62E, platelet-derived microparticles with CD62P and CD42b, leukocyte-derived microparticles with CD45 and monocyte-derived microparticles with CD14. Results Marathon running induced leukocytosis (5.9 ± 0.1 to 14.8 ± 0.3 10 9 /l, p < 0.0001) and increased platelet counts (239 ± 4.6 to 281 ± 5.9 10 9 /l, p < 0.0001) immediately after the marathon. Blood monocytes increased and lymphocytes decreased after the run ( p < 0.0001). Endothelial-derived microparticles were acutely increased ( p = 0.008) due to a 23% increase of apoptotic endothelial-derived microparticles ( p = 0.007) and returned to baseline within two days after the marathon. Thrombocyte-derived microparticles acutely increased by 38% accompanied by an increase in activated and apoptotic thrombocyte-derived microparticles ( p ≤ 0.0001) each. Both monocyte- and leukocyte-derived microparticles were decreased immediately after marathon run ( p < 0.0001) and remained below baseline until day 2. Troponin T increased from 12 to 32 ng/l ( p < 0.0001) immediately after the run and returned to baseline after two days. Conclusion Circulating apoptotic endothelial- and thrombocyte-derived microparticles increased after marathon running consistent with an acute pro-thrombotic and pro-inflammatory state. Exercise-induced vascular damage reflected by microparticles could indicate potential mechanisms of post-exertional cardiovascular complications. Further

  6. Chain Rule Approach for Calculating the Time-Derivative of Flux

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

    Langenbrunner, James R.; Booker, Jane M.

    The reaction history (gamma-flux observable) is mathematically studied by using the chain rule for taking the total-time derivatives. That is, the total time-derivative of flux is written as the product of the ion temperature derivative with respect to time and the derivative of the flux with respect to ion temperature. Some equations are derived using the further simplification that the fusion reactivity is a parametrized function of ion temperature, T. Deuterium-tritium (D-T) fusion is used as the application with reactivity calculations from three established reactivity parametrizations.

  7. Synthesis and characterization of bis-thiourea having amino acid derivatives

    NASA Astrophysics Data System (ADS)

    Fakhar, Imran; Yamin, Bohari M.; Hasbullah, Siti Aishah

    2016-11-01

    In this article four new symmetric bis-thiourea derivatives having amino acid linkers were reported with good yield. Isophthaloyl dichloride was used as spacer and L-alanine, L-aspartic acid, L-phenylalanine and L-glutamic acid were used as linkers. Bis-thiourea derivatives were prepared from relatively stable isophthaloyl isothiocyanate intermediate. Newly synthesized bis-thiourea derivatives were characterized by FTIR, H-NMR, 13C-NMR and CHNS-O elemental analysis techniques. Characterization data was in good agreement with the expected derivatives, hence confirmed the synthesis of four new derivatives of bis-thiourea having amino acids.

  8. 12 CFR 217.34 - OTC derivative contracts.

    Code of Federal Regulations, 2014 CFR

    2014-01-01

    ... net present value of the amount of unpaid premiums. Table 1 to § 217.34—Conversion Factor Matrix for... single OTC derivative contract is the greater of the mark-to-fair value of the OTC derivative contract or... with a negative mark-to-fair value, is calculated by multiplying the notional principal amount of the...

  9. 12 CFR 3.34 - OTC derivative contracts.

    Code of Federal Regulations, 2014 CFR

    2014-01-01

    ..., notional principal amount is the net receipts to each party falling due on each value date in each currency... calculating PFE. (E) The PFE of the protection provider of a credit derivative is capped at the net present... the greater of the mark-to-fair value of the OTC derivative contract or zero. (ii) PFE. (A) The PFE...

  10. 21 CFR 184.1685 - Rennet (animal-derived) and chymosin preparation (fermentation-derived).

    Code of Federal Regulations, 2012 CFR

    2012-04-01

    ... commercial extracts containing the active enzyme rennin (CAS Reg. No. 9001-98-3), also known as chymosin (International Union of Biochemistry Enzyme Commission (E.C.) 3.4.23.4). Rennet is the aqueous extract prepared... clear solution containing the active enzyme chymosin (E.C. 3.4.23.4). It is derived, via fermentation...

  11. 21 CFR 184.1685 - Rennet (animal-derived) and chymosin preparation (fermentation-derived).

    Code of Federal Regulations, 2013 CFR

    2013-04-01

    ... commercial extracts containing the active enzyme rennin (CAS Reg. No. 9001-98-3), also known as chymosin (International Union of Biochemistry Enzyme Commission (E.C.) 3.4.23.4). Rennet is the aqueous extract prepared... clear solution containing the active enzyme chymosin (E.C. 3.4.23.4). It is derived, via fermentation...

  12. 21 CFR 184.1685 - Rennet (animal-derived) and chymosin preparation (fermentation-derived).

    Code of Federal Regulations, 2014 CFR

    2014-04-01

    ... active enzyme rennin (CAS Reg. No. 9001-98-3), also known as chymosin (International Union of Biochemistry Enzyme Commission (E.C.) 3.4.23.4). Rennet is the aqueous extract prepared from cleaned, frozen... active enzyme chymosin (E.C. 3.4.23.4). It is derived, via fermentation, from a nonpathogenic and...

  13. Geochemical Constraints for Mercury's PCA-Derived Geochemical Terranes

    NASA Astrophysics Data System (ADS)

    Stockstill-Cahill, K. R.; Peplowski, P. N.

    2018-05-01

    PCA-derived geochemical terranes provide a robust, analytical means of defining these terranes using strictly geochemical inputs. Using the end members derived in this way, we are able to assess the geochemical implications for Mercury.

  14. Acclimation of aerobic-activated sludge degrading benzene derivatives and co-metabolic degradation activities of trichloroethylene by benzene derivative-grown aerobic sludge.

    PubMed

    Wang, Shizong; Yang, Qi; Bai, Zhiyong; Wang, Shidong; Wang, Yeyao; Nowak, Karolina M

    2015-01-01

    The acclimation of aerobic-activated sludge for degradation of benzene derivatives was investigated in batch experiments. Phenol, benzoic acid, toluene, aniline and chlorobenzene were concurrently added to five different bioreactors which contained the aerobic-activated sludge. After the acclimation process ended, the acclimated phenol-, benzoic acid-, toluene-, aniline- and chlorobenzene-grown aerobic-activated sludge were used to explore the co-metabolic degradation activities of trichloroethylene (TCE). Monod equation was employed to simulate the kinetics of co-metabolic degradation of TCE by benzene derivative-grown sludge. At the end of experiments, the mixed microbial communities grown under different conditions were identified. The results showed that the acclimation periods of microorganisms for different benzene derivatives varied. The maximum degradation rates of TCE for phenol-, benzoic acid-, toluene-, aniline- and chlorobenzene-grown aerobic sludge were 0.020, 0.017, 0.016, 0.0089 and 0.0047 mg g SS(-1) h(-1), respectively. The kinetic of TCE degradation in the absence of benzene derivative followed Monod equation well. Also, eight phyla were observed in the acclimated benzene derivative-grown aerobic sludge. Each of benzene derivative-grown aerobic sludge had different microbial community composition. This study can hopefully add new knowledge to the area of TCE co-metabolic by mixed microbial communities, and further the understanding on the function and applicability of aerobic-activated sludge.

  15. Holographic superconductivity from higher derivative theory

    NASA Astrophysics Data System (ADS)

    Wu, Jian-Pin; Liu, Peng

    2017-11-01

    We construct a 6 derivative holographic superconductor model in the 4-dimensional bulk spacetimes, in which the normal state describes a quantum critical (QC) phase. The phase diagram (γ1 ,Tˆc) and the condensation as the function of temperature are worked out numerically. We observe that with the decrease of the coupling parameter γ1, the critical temperature Tˆc decreases and the formation of charged scalar hair becomes harder. We also calculate the optical conductivity. An appealing characteristic is a wider extension of the superconducting energy gap, comparing with that of 4 derivative theory. It is expected that this phenomena can be observed in the real materials of high temperature superconductor. Also the Homes' law in our present models with 4 and 6 derivative corrections is explored. We find that in certain range of parameters γ and γ1, the experimentally measured value of the universal constant C in Homes' law can be obtained.

  16. Cinnamic acid and its derivatives inhibit fructose-mediated protein glycation.

    PubMed

    Adisakwattana, Sirichai; Sompong, Weerachat; Meeprom, Aramsri; Ngamukote, Sathaporn; Yibchok-Anun, Sirintorn

    2012-01-01

    Cinnamic acid and its derivatives have shown a variety of pharmacologic properties. However, little is known about the antiglycation properties of cinnamic acid and its derivatives. The present study sought to characterize the protein glycation inhibitory activity of cinnamic acid and its derivatives in a bovine serum albumin (BSA)/fructose system. The results demonstrated that cinnamic acid and its derivatives significantly inhibited the formation of advanced glycation end products (AGEs) by approximately 11.96-63.36% at a concentration of 1 mM. The strongest inhibitory activity against the formation of AGEs was shown by cinnamic acid. Furthermore, cinnamic acid and its derivatives reduced the level of fructosamine, the formation of N(ɛ)-(carboxymethyl) lysine (CML), and the level of amyloid cross β-structure. Cinnamic acid and its derivatives also prevented oxidative protein damages, including effects on protein carbonyl formation and thiol oxidation of BSA. Our findings may lead to the possibility of using cinnamic acid and its derivatives for preventing AGE-mediated diabetic complications.

  17. Cinnamic Acid and Its Derivatives Inhibit Fructose-Mediated Protein Glycation

    PubMed Central

    Adisakwattana, Sirichai; Sompong, Weerachat; Meeprom, Aramsri; Ngamukote, Sathaporn; Yibchok-anun, Sirintorn

    2012-01-01

    Cinnamic acid and its derivatives have shown a variety of pharmacologic properties. However, little is known about the antiglycation properties of cinnamic acid and its derivatives. The present study sought to characterize the protein glycation inhibitory activity of cinnamic acid and its derivatives in a bovine serum albumin (BSA)/fructose system. The results demonstrated that cinnamic acid and its derivatives significantly inhibited the formation of advanced glycation end products (AGEs) by approximately 11.96–63.36% at a concentration of 1 mM. The strongest inhibitory activity against the formation of AGEs was shown by cinnamic acid. Furthermore, cinnamic acid and its derivatives reduced the level of fructosamine, the formation of Nɛ-(carboxymethyl) lysine (CML), and the level of amyloid cross β-structure. Cinnamic acid and its derivatives also prevented oxidative protein damages, including effects on protein carbonyl formation and thiol oxidation of BSA. Our findings may lead to the possibility of using cinnamic acid and its derivatives for preventing AGE-mediated diabetic complications. PMID:22408423

  18. 5S rRNA-derived and tRNA-derived SINEs in fruit bats.

    PubMed

    Gogolevsky, Konstantin P; Vassetzky, Nikita S; Kramerov, Dmitri A

    2009-05-01

    Most short retroposons (SINEs) descend from cellular tRNA of 7SL RNA. Here, four new SINEs were found in megabats (Megachiroptera) but neither in microbats nor in other mammals. Two of them, MEG-RS and MEG-RL, descend from another cellular RNA, 5S rRNA; one (MEG-T2) is a tRNA-derived SINE; and MEG-TR is a hybrid tRNA/5S rRNA SINE. Insertion locus analysis suggests that these SINEs were active in the recent fruit bat evolution. Analysis of MEG-RS and MEG-RL in comparison with other few 5S rRNA-derived SINEs demonstrates that the internal RNA polymerase III promoter is their most invariant region, while the secondary structure is more variable. The mechanisms underlying the modular structure of these and other SINEs as well as their variation are discussed. The scenario of evolution of MEG SINEs is proposed.

  19. Synthesis of 5'-deoxy-5'-nucleosideacetic acid derivatives

    NASA Technical Reports Server (NTRS)

    Harada, Kazuo; Orgel, Leslie E.

    1990-01-01

    Several new 5'-deoxy-5'-nucleosideacetic acid derivatives have been synthesized by the reactions of alkoxycarbonylmethylene triphenylphosphoranes with nucleoside 5'-aldehydes. The oligomerization of adenine derivatives IIa, IIIa, IV, V and guanine derivatives IIc and IIIc in aqueous solution was studied using a water-soluble carbodiimide as a condensing agent. It is found that the saturated acid (IV) tends to cyclize to the lactone, while IIa and unsaturated acids (IIIa and V) oligomerized efficiently, especially in the presence of poly (U) as a template.

  20. Second derivative in the model of classical binary system

    NASA Astrophysics Data System (ADS)

    Abubekerov, M. K.; Gostev, N. Yu.

    2016-06-01

    We have obtained an analytical expression for the second derivatives of the light curve with respect to geometric parameters in the model of eclipsing classical binary systems. These expressions are essentially efficient algorithm to calculate the numerical values of these second derivatives for all physical values of geometric parameters. Knowledge of the values of second derivatives of the light curve at some point provides additional information about asymptotical behaviour of the function near this point and can significantly improve the search for the best-fitting light curve through the use of second-order optimization method. We write the expression for the second derivatives in a form which is most compact and uniform for all values of the geometric parameters and so make it easy to write a computer program to calculate the values of these derivatives.

  1. In defense of derivations

    NASA Astrophysics Data System (ADS)

    Mungan, Carl E.

    2016-05-01

    At the 2015 AAPT Summer Meeting, I presented four derivations of the formula for motional emf. Such physics derivations involve the construction of explanatory frameworks involving diagrams and mathematical models. Although textbooks devote considerable space to such explanations, many teachers and students spend their time on worksheets, end-of-chapter problems, and the like. The book is reduced to a bank of solved (i.e., example) and unsolved (i.e., homework) questions, along with equations in colored boxes that presumably are to be used to answer those questions. Such an approach encourages fragmentation of knowledge, the view that there is only one right answer to a problem with the goal of physics being to find that answer (neatly boxed of course), and the inability to reason about even a slightly different (much less a novel) situation. If we are to develop scientific literacy, significant course time must be devoted to explaining the structure of and support for the models and equations we use.

  2. High precision pulsar timing and spin frequency second derivatives

    NASA Astrophysics Data System (ADS)

    Liu, X. J.; Bassa, C. G.; Stappers, B. W.

    2018-05-01

    We investigate the impact of intrinsic, kinematic and gravitational effects on high precision pulsar timing. We present an analytical derivation and a numerical computation of the impact of these effects on the first and second derivative of the pulsar spin frequency. In addition, in the presence of white noise, we derive an expression to determine the expected measurement uncertainty of a second derivative of the spin frequency for a given timing precision, observing cadence and timing baseline and find that it strongly depends on the latter (∝t-7/2). We show that for pulsars with significant proper motion, the spin frequency second derivative is dominated by a term dependent on the radial velocity of the pulsar. Considering the data sets from three Pulsar Timing Arrays, we find that for PSR J0437-4715 a detectable spin frequency second derivative will be present if the absolute value of the radial velocity exceeds 33 km s-1. Similarly, at the current timing precision and cadence, continued timing observations of PSR J1909-3744 for about another eleven years, will allow the measurement of its frequency second derivative and determine the radial velocity with an accuracy better than 14 km s-1. With the ever increasing timing precision and observing baselines, the impact of the, largely unknown, radial velocities of pulsars on high precision pulsar timing can not be neglected.

  3. Deriving stellar parameters with the SME software package

    NASA Astrophysics Data System (ADS)

    Piskunov, N.

    2017-09-01

    Photometry and spectroscopy are complementary tools for deriving accurate stellar parameters. Here I present one of the popular packages for stellar spectroscopy called SME with the emphasis on the latest developments and error assessment for the derived parameters.

  4. Kojyl cinnamate ester derivatives promote adiponectin production during adipogenesis in human adipose tissue-derived mesenchymal stem cells.

    PubMed

    Rho, Ho Sik; Hong, Soo Hyun; Park, Jongho; Jung, Hyo-Il; Park, Young-Ho; Lee, John Hwan; Shin, Song Seok; Noh, Minsoo

    2014-05-01

    The subcutaneous fat tissue mass gradually decreases with age, and its regulation is a strategy to develop anti-aging compounds to ameliorate the photo-aging of human skin. The adipogenesis of human adipose tissue-mesenchymal stem cells (hAT-MSCs) can be used as a model to discover novel anti-aging compounds. Cinnamomum cassia methanol extracts were identified as adipogenesis-promoting agents by natural product library screening. Cinnamates, the major chemical components of Cinnamomum cassia extracts, promoted adipogenesis in hAT-MSCs. We synthesized kojyl cinnamate ester derivatives to improve the pharmacological activity of cinnamates. Structure-activity studies of kojyl cinnamate derivatives showed that both the α,β-unsaturated carbonyl ester group and the kojic acid moiety play core roles in promoting adiponectin production during adipogenesis in hAT-MSCs. We conclude that kojyl cinnamate ester derivatives provide novel pharmacophores that can regulate adipogenesis in hAT-MSCs. Copyright © 2014 Elsevier Ltd. All rights reserved.

  5. Structure-activity relationships among substituted N-benzoyl derivatives of phenylalanine and its analogues in a microbial antitumor prescreen III: derivatives of p-fluoro-DL-phenylalanine.

    PubMed

    Otani, T T; Briley, M R

    1985-01-01

    Twelve substituted benzoyl derivatives of p-fluoro-DL-phenylalanine were prepared and tested for growth-inhibitory activity in a Lactobacillus casei system used as an antitumor prescreen. The 12 substituted benzoyl groups were the same as those attached to o-fluorophenylalanine and m-fluorophenylalanine studied earlier. The activity of these compounds was compared vertically among themselves and horizontally with the corresponding derivatives of o-fluorophenylalanine and of m-fluorophenylalanine. It was found that the derivatives of p-fluorophenylalanine, like those of o- and m-fluorophenylalanine, exhibited remarkable inhibition, all but one, i.e., the o-nitrobenzoyl derivative, showing inhibition that is considered to be positive in the prescreen. Particularly potent compounds in this group were the m-chlorobenzoyl-, p-chlorobenzoyl, m-nitrobenzoyl, and p-nitrobenzoyl derivatives. Comparison of the activity of the substituted benzoyl derivatives of all three structural isomers of fluorophenylalanine at equimolar concentrations showed that the derivatives of m-fluorophenylalanine were generally better inhibitors than those of o-fluoro- or p-fluorophenylalanine. Study of the ID50 values of the more active substituted benzoyl derivatives of the fluorophenylalanines showed that the most active of this group was m-chlorobenzoyl-p-fluoro-DL-phenylalanine.

  6. Photoelectrochemical hydrogen production from biomass derivatives and water.

    PubMed

    Lu, Xihong; Xie, Shilei; Yang, Hao; Tong, Yexiang; Ji, Hongbing

    2014-11-21

    Hydrogen, a clean energy carrier with high energy capacity, is a very promising candidate as a primary energy source for the future. Photoelectrochemical (PEC) hydrogen production from renewable biomass derivatives and water is one of the most promising approaches to producing green chemical fuel. Compared to water splitting, hydrogen production from renewable biomass derivatives and water through a PEC process is more efficient from the viewpoint of thermodynamics. Additionally, the carbon dioxide formed can be re-transformed into carbohydrates via photosynthesis in plants. In this review, we focus on the development of photoanodes and systems for PEC hydrogen production from water and renewable biomass derivatives, such as methanol, ethanol, glycerol and sugars. We also discuss the future challenges and opportunities for the design of the state-of-the-art photoanodes and PEC systems for hydrogen production from biomass derivatives and water.

  7. Progress on coal-derived fuels for aviation systems

    NASA Technical Reports Server (NTRS)

    Witcofski, R. D.

    1978-01-01

    Synthetic aviation kerosene (Syn. Jet-A), liquid methane (LCH4), and liquid hydrogen (LH2) appear to be the most promising coal-derived fuels. Liquid hydrogen aircraft configurations, their fuel systems, and their ground requirements at the airport are identified. These aircraft appear viable, particularly for long haul use, where aircraft fueled with coal derived LH2 would consume 9 percent less coal resources than would aircraft fueled with coal derived Syn. Jet-A. Distribution of hydrogen from the point of manufacture to airports may pose problems. Synthetic JET-A would appear to cause fewer concerns to the air transportation industry. Of the three candidate fuels, LCH4 is the most energy efficient to produce, and an aircraft fueled with coal derived LCH4 may provide both the most efficient utilization of coal resources and the least expensive ticket as well.

  8. Derivative expansion of wave function equivalent potentials

    NASA Astrophysics Data System (ADS)

    Sugiura, Takuya; Ishii, Noriyoshi; Oka, Makoto

    2017-04-01

    Properties of the wave function equivalent potentials introduced by the HAL QCD collaboration are studied in a nonrelativistic coupled-channel model. The derivative expansion is generalized, and then applied to the energy-independent and nonlocal potentials. The expansion coefficients are determined from analytic solutions to the Nambu-Bethe-Salpeter wave functions. The scattering phase shifts computed from these potentials are compared with the exact values to examine the convergence of the expansion. It is confirmed that the generalized derivative expansion converges in terms of the scattering phase shift rather than the functional structure of the non-local potentials. It is also found that the convergence can be improved by tuning either the choice of interpolating fields or expansion scale in the generalized derivative expansion.

  9. Applications of several spectral techniques to characterize coordination compounds derived from 2,6-diacetylpyridine derivative

    NASA Astrophysics Data System (ADS)

    Chandra, Sulekh; Sharma, Amit Kumar

    2009-09-01

    The coordination compounds of Cr III, Mn II and Co II metal ions derived from quinquedentate 2,6-diacetylpyridine derivative have been synthesized and characterized by using the various physicochemical studies like stoichiometric, molar conductivity and magnetic, and spectral techniques like IR, NMR, mass, UV and EPR. The general stoichiometries of the complexes are found to be [Cr(H 2L)X] and [M(HL)X], where M = Mn(II) and Co(II); H 2L = dideprotonated ligand, HL = monodeprotonated ligand and X = NO 3-, Cl - and OAc -. The studies reveal that the complexes possess monomeric compositions with six coordinated octahedral geometry (Cr III and Mn II complexes) and six coordinated tetragonal geometry (Co II complexes).

  10. The susceptibility of the retina to photochemical damage from visible light

    PubMed Central

    Hunter, Jennifer J; Morgan, Jessica I W; Merigan, William H; Sliney, David H; Sparrow, Janet R; Williams, David R

    2011-01-01

    The photoreceptor/RPE complex must maintain a delicate balance between maximizing the absorption of photons for vision and retinal image quality while simultaneously minimizing the risk of photodamage when exposed to bright light. We review the recent discovery of two new effects of light exposure on the photoreceptor/RPE complex in the context of current thinking about the causes of retinal phototoxicity. These effects are autofluorescence photobleaching in which exposure to bright light reduces lipofuscin autofluorescence and, at higher light levels, RPE disruption in which the pattern of autofluorescence is permanently altered following light exposure. Both effects occur following exposure to visible light at irradiances that were previously thought to be safe. Photopigment, retinoids involved in the visual cycle, and bisretinoids in lipofuscin have been implicated as possible photosensitizers for photochemical damage. The mechanism of RPE disruption may follow either of these paths. On the other hand, autofluorescence photobleaching is likely an indicator of photooxidation of lipofuscin. The permanent changes inherent in RPE disruption might require modification of the light safety standards. AF photobleaching recovers after several hours although the mechanisms by which this occurs are not yet clear. Understanding the mechanisms of phototoxicity is all the more important given the potential for increased susceptibility in the presence of ocular diseases that affect either the visual cycle and/or lipofuscin accumulation. In addition, knowledge of photochemical mechanisms can improve our understanding of some disease processes that may be influenced by light exposure, such as some forms of Leber’s congenital amaurosis, and aid in the development of new therapies. Such treatment prior to intentional light exposures, as in ophthalmic examinations or surgeries, could provide an effective preventative strategy. PMID:22085795

  11. Translational Diffusion of Macromolecule-sized Solutes in Cytoplasm and Nucleus

    PubMed Central

    Seksek, Olivier; Biwersi, Joachim; Verkman, A.S.

    1997-01-01

    Fluorescence recovery after photobleaching (FRAP) was used to quantify the translational diffusion of microinjected FITC-dextrans and Ficolls in the cytoplasm and nucleus of MDCK epithelial cells and Swiss 3T3 fibroblasts. Absolute diffusion coefficients (D) were measured using a microsecond-resolution FRAP apparatus and solution standards. In aqueous media (viscosity 1 cP), D for the FITC-dextrans decreased from 75 to 8.4 × 10−7 cm2/s with increasing dextran size (4–2,000 kD). D in cytoplasm relative to that in water (D/Do) was 0.26 ± 0.01 (MDCK) and 0.27 ± 0.01 (fibroblasts), and independent of FITC-dextran and Ficoll size (gyration radii [RG] 40–300 Å). The fraction of mobile FITC-dextran molecules (fmob), determined by the extent of fluorescence recovery after spot photobleaching, was >0.75 for RG < 200 Å, but decreased to <0.5 for RG > 300 Å. The independence of D/Do on FITC-dextran and Ficoll size does not support the concept of solute “sieving” (size-dependent diffusion) in cytoplasm. Photobleaching measurements using different spot diameters (1.5–4 μm) gave similar D/Do, indicating that microcompartments, if present, are of submicron size. Measurements of D/Do and fmob in concentrated dextran solutions, as well as in swollen and shrunken cells, suggested that the low fmob for very large macromolecules might be related to restrictions imposed by immobile obstacles (such as microcompartments) or to anomalous diffusion (such as percolation). In nucleus, D/Do was 0.25 ± 0.02 (MDCK) and 0.27 ± 0.03 (fibroblasts), and independent of solute size (RG 40–300 Å). Our results indicate relatively free and rapid diffusion of macromolecule-sized solutes up to approximately 500 kD in cytoplasm and nucleus. PMID:9214387

  12. Synthesis of 4,19-disubstituted derivatives of DOC. Radioreceptor assay of some corticosteroid derivatives in human mononuclear leukocytes.

    PubMed

    Harnik, M; Kashman, Y; Cojocaru, M; Bauer, H; Laux, M; Lewicka, S; Vecsei, P

    1990-10-01

    Several new 4,19-substituted steroids and previously synthesized corticosteroids were assayed for affinity to type 1 receptors in human mononuclear leukocytes. 11 beta,19-epoxy-4,21-dihydroxypregn-4-ene-3,20-dione (2) was hydrogenated with Pd-C to yield a mixture of all four dihydro derivatives 5, accompanied by 4,21-diacetoxy-11 beta,19-epoxy-3-hydroxypregnan-20-one (6) and 21-acetoxy-11 beta,19-epoxy-4-hydroxypregnane-3,20-dione (7). With hot acetic + p-toluenesulfonic acid 5 underwent rearrangement to 21-acetoxy-11 beta,19-epoxypregn-5-ene-4,20-dione (8) Pd-C hydrogenation of 3,21-diacetoxy-5 beta,19-cyclopregna-2,9(11)-diene-4,20-dione (10) gave 3,21-diacetoxy-5 beta,19-cyclopregn-5-ene-4,20-dione (11) and the 9,11-dihydro derivative of the latter. Treatment of 10 with warm HCl furnished 19-chloro-4,21-dihydroxypregna-4,9(11)-diene-3,20-dione (13). Pd-C hydrogenation of its diacetate 14 afforded the 4,5-dihydro derivative 18, 19-chloro-21-acetoxypregn-9(11)-en-20-one (15), its 4-acetoxy derivative 16 and the 3,4-diacetoxy derivative 17. When tested in a radioreceptor assay in human mononuclear leukocytes the synthesized compounds showed only low relative binding affinities (RBA) to type 1 receptor, the highest being 0.72% for 13 (aldosterone = 100%). For comparison, other RBA in this system were: 19-noraldosterone, 20%; 18-deoxyaldosterone, 5.8%; 18-deoxy-19-noraldosterone, 4.7%; 18,21-anhydroaldosterone, 0.37%; 17-isoaldosterone, 7.6% and apoaldosterone, 4.3%

  13. Cohesin Can Remain Associated with Chromosomes during DNA Replication.

    PubMed

    Rhodes, James D P; Haarhuis, Judith H I; Grimm, Jonathan B; Rowland, Benjamin D; Lavis, Luke D; Nasmyth, Kim A

    2017-09-19

    To ensure disjunction to opposite poles during anaphase, sister chromatids must be held together following DNA replication. This is mediated by cohesin, which is thought to entrap sister DNAs inside a tripartite ring composed of its Smc and kleisin (Scc1) subunits. How such structures are created during S phase is poorly understood, in particular whether they are derived from complexes that had entrapped DNAs prior to replication. To address this, we used selective photobleaching to determine whether cohesin associated with chromatin in G1 persists in situ after replication. We developed a non-fluorescent HaloTag ligand to discriminate the fluorescence recovery signal from labeling of newly synthesized Halo-tagged Scc1 protein (pulse-chase or pcFRAP). In cells where cohesin turnover is inactivated by deletion of WAPL, Scc1 can remain associated with chromatin throughout S phase. These findings suggest that cohesion might be generated by cohesin that is already bound to un-replicated DNA. Copyright © 2017 The Author(s). Published by Elsevier Inc. All rights reserved.

  14. The escape problem for mortal walkers

    NASA Astrophysics Data System (ADS)

    Grebenkov, D. S.; Rupprecht, J.-F.

    2017-02-01

    We introduce and investigate the escape problem for random walkers that may eventually die, decay, bleach, or lose activity during their diffusion towards an escape or reactive region on the boundary of a confining domain. In the case of a first-order kinetics (i.e., exponentially distributed lifetimes), we study the effect of the associated death rate onto the survival probability, the exit probability, and the mean first passage time. We derive the upper and lower bounds and some approximations for these quantities. We reveal three asymptotic regimes of small, intermediate, and large death rates. General estimates and asymptotics are compared to several explicit solutions for simple domains and to numerical simulations. These results allow one to account for stochastic photobleaching of fluorescent tracers in bio-imaging, degradation of mRNA molecules in genetic translation mechanisms, or high mortality rates of spermatozoa in the fertilization process. Our findings provide a mathematical ground for optimizing storage containers and materials to reduce the risk of leakage of dangerous chemicals or nuclear wastes.

  15. Single lipid bilayer deposition on polymer surfaces using bicelles.

    PubMed

    Saleem, Qasim; Zhang, Zhenfu; Petretic, Amy; Gradinaru, Claudiu C; Macdonald, Peter M

    2015-03-09

    A lipid bilayer was deposited on a 3 μm diameter polystyrene (PS) bead via hydrophobic anchoring of bicelles containing oxyamine-bearing cholesteric moieties reacting with the aldehyde functionalized bead surface. Discoidal bicelles were formed by mixing dimyristoylphosphatidylcholine (DMPC), dihexanoylphosphatidylcholine (DHPC), dimyristoyltrimethylammonium propane (DMTAP), and the oxyamine-terminated cholesterol derivative, cholest-5-en-3β-oxy-oct-3,6-oxa-an-8-oxyamine (CHOLOA), in the molar ratio DMPC/DHCP/DMTAP/CHOLOA (1/0.5/0.01/0.05) in water. Upon exposure to aldehyde-bearing PS beads, a stable single lipid bilayer coating rapidly formed at the bead surface. Fluorescence recovery after photobleaching demonstrated that the deposited lipids fused into an encapsulating lipid bilayer. Electrospray ionization mass spectrometry showed that the short chain lipid DHPC was entirely absent from the PS adherent lipid coating. Fluorescence quenching measurements proved that the coating was a single lipid bilayer. The bicelle coating method is thus simple and robust, can be modified to include membrane-associated species, and can be adapted to coat any number of different surfaces.

  16. Preparation of highly luminescent and color tunable carbon nanodots under visible light excitation for in vitro and in vivo bio-imaging

    DOE PAGES

    Zheng, Min; Li, Jing; Xie, Zhigang; ...

    2015-11-10

    Carbon nanodots (CDs) have generated enormous excitement because of their superiority in water solubility, chemical inertness, low toxicity, ease of functionalization and resistance to photobleaching. Here we report a facile thermal pyrolysis route to prepare CDs with high quantum yield (QY) using citric acid as the carbon source and ethylene diamine derivatives (EDAs) including triethylenetetramine (TETA), tetraethylenepentamine (TEPA) and polyene polyamine (PEPA) as the passivation agents. We find that the CDs prepared from EDAs, such as TETA, TEPA and PEPA, show relatively high photoluminescence (PL) QY (11.4, 10.6, and 9.8%, respectively) at 1 ex of 465 nm. The cytotoxicity ofmore » the CDs has been investigated through in vitro and in vivo bio-imaging studies. The results indicate that these CDs possess low toxicity and good biocompatibility. As a result, the unique properties such as the high PL QY at large excitation wave length and the low toxicity of the resulting CDs make them promising fluorescent nanoprobes for applications in optical bio-imaging and biosensing.« less

  17. Ionophore-based optical nanosensors incorporating hydrophobic carbon dots and a pH-sensitive quencher dye for sodium detection.

    PubMed

    Galyean, A A; Behr, M R; Cash, K J

    2018-01-21

    Nanosensors present a biological monitoring method that is biocompatible, reversible, and nano-scale, and they offer many advantages over traditional organic indicators. Typical ionophore-based nanosensors incorporate nile-blue derivative pH indicators but suffer from photobleaching while quantum dot alternatives pose a potential toxicity risk. In order to address this challenge, sodium selective nanosensors containing carbon dots and a pH-sensitive quencher molecule were developed based on an ion-exchange theory and a decoupled recognition element from the pH indicator. Carbon dots were synthesized and integrated into nanosensors containing a pH-indicator, an analyte-binding ligand (ionophore), and a charge-balancing additive. These nanosensors are ion-selective against potassium (selectivity coefficient of 0.4) and lithium (selectivity coefficient of 0.9). Reversible nanosensor response to sodium is also demonstrated. The carbon dot nanosensors are resistant to changes in optical properties for at least 12 h and display stable selectivity to physiologically-relevant sodium (alpha = 0.5 of 200 mM NaCl) for a minimum of 6 days.

  18. Intraspinal Rewiring of the Corticospinal Tract Requires Target-Derived Brain-Derived Neurotrophic Factor and Compensates Lost Function after Brain Injury

    ERIC Educational Resources Information Center

    Ueno, Masaki; Hayano, Yasufumi; Nakagawa, Hiroshi; Yamashita, Toshihide

    2012-01-01

    Brain injury that results in an initial behavioural deficit is frequently followed by spontaneous recovery. The intrinsic mechanism of this functional recovery has never been fully understood. Here, we show that reorganization of the corticospinal tract induced by target-derived brain-derived neurotrophic factor is crucial for spontaneous recovery…

  19. Transgene Reactivation in Induced Pluripotent Stem Cell Derivatives and Reversion to Pluripotency of Induced Pluripotent Stem Cell-Derived Mesenchymal Stem Cells

    PubMed Central

    Galat, Yekaterina; Perepitchka, Mariana; Jennings, Lawrence J.; Iannaccone, Philip M.; Hendrix, Mary J.C.

    2016-01-01

    Induced pluripotent stem cells (iPSCs) have enormous potential in regenerative medicine and disease modeling. It is now felt that clinical trials should be performed with iPSCs derived with nonintegrative constructs. Numerous studies, however, including those describing disease models, are still being published using cells derived from iPSCs generated with integrative constructs. Our experimental work presents the first evidence of spontaneous transgene reactivation in vitro in several cellular types. Our results show that the transgenes were predominantly silent in parent iPSCs, but in mesenchymal and endothelial iPSC derivatives, the transgenes experienced random upregulation of Nanog and c-Myc. Additionally, we provide evidence of spontaneous secondary reprogramming and reversion to pluripotency in mesenchymal stem cells derived from iPSCs. These findings strongly suggest that the studies, which use cellular products derived from iPSCs generated with retro- or lentiviruses, should be evaluated with consideration of the possibility of transgene reactivation. The in vitro model described here provides insight into the earliest events of culture transformation and suggests the hypothesis that reversion to pluripotency may be responsible for the development of tumors in cell replacement experiments. The main goal of this work, however, is to communicate the possibility of transgene reactivation in retro- or lenti-iPSC derivatives and the associated loss of cellular fidelity in vitro, which may impact the outcomes of disease modeling and related experimentation. PMID:27193052

  20. Special Relativity Derived from Spacetime Magma

    PubMed Central

    Greensite, Fred

    2014-01-01

    We present a derivation of relativistic spacetime largely untethered from specific physical considerations, in constrast to the many physically-based derivations that have appeared in the last few decades. The argument proceeds from the inherent magma (groupoid) existing on the union of spacetime frame components and Euclidean which is consistent with an “inversion symmetry” constraint from which the Minkowski norm results. In this context, the latter is also characterized as one member of a class of “inverse norms” which play major roles with respect to various unital -algebras more generally. PMID:24959889

  1. Enabling quaternion derivatives: the generalized HR calculus

    PubMed Central

    Xu, Dongpo; Jahanchahi, Cyrus; Took, Clive C.; Mandic, Danilo P.

    2015-01-01

    Quaternion derivatives exist only for a very restricted class of analytic (regular) functions; however, in many applications, functions of interest are real-valued and hence not analytic, a typical case being the standard real mean square error objective function. The recent HR calculus is a step forward and provides a way to calculate derivatives and gradients of both analytic and non-analytic functions of quaternion variables; however, the HR calculus can become cumbersome in complex optimization problems due to the lack of rigorous product and chain rules, a consequence of the non-commutativity of quaternion algebra. To address this issue, we introduce the generalized HR (GHR) derivatives which employ quaternion rotations in a general orthogonal system and provide the left- and right-hand versions of the quaternion derivative of general functions. The GHR calculus also solves the long-standing problems of product and chain rules, mean-value theorem and Taylor's theorem in the quaternion field. At the core of the proposed GHR calculus is quaternion rotation, which makes it possible to extend the principle to other functional calculi in non-commutative settings. Examples in statistical learning theory and adaptive signal processing support the analysis. PMID:26361555

  2. Enabling quaternion derivatives: the generalized HR calculus.

    PubMed

    Xu, Dongpo; Jahanchahi, Cyrus; Took, Clive C; Mandic, Danilo P

    2015-08-01

    Quaternion derivatives exist only for a very restricted class of analytic (regular) functions; however, in many applications, functions of interest are real-valued and hence not analytic, a typical case being the standard real mean square error objective function. The recent HR calculus is a step forward and provides a way to calculate derivatives and gradients of both analytic and non-analytic functions of quaternion variables; however, the HR calculus can become cumbersome in complex optimization problems due to the lack of rigorous product and chain rules, a consequence of the non-commutativity of quaternion algebra. To address this issue, we introduce the generalized HR (GHR) derivatives which employ quaternion rotations in a general orthogonal system and provide the left- and right-hand versions of the quaternion derivative of general functions. The GHR calculus also solves the long-standing problems of product and chain rules, mean-value theorem and Taylor's theorem in the quaternion field. At the core of the proposed GHR calculus is quaternion rotation, which makes it possible to extend the principle to other functional calculi in non-commutative settings. Examples in statistical learning theory and adaptive signal processing support the analysis.

  3. Antihypertensive properties of lactoferricin B-derived peptides.

    PubMed

    Ruiz-Giménez, Pedro; Ibáñez, Aida; Salom, Juan B; Marcos, Jose F; López-Díez, Jose Javier; Vallés, Salvador; Torregrosa, Germán; Alborch, Enrique; Manzanares, Paloma

    2010-06-09

    A set of eight lactoferricin B (LfcinB)-derived peptides was examined for inhibitory effects on angiotensin I-converting enzyme (ACE) activity and ACE-dependent vasoconstriction, and their hypotensive effect in spontaneously hypertensive rats (SHR). Peptides were derived from different elongations both at the C-terminal and N-terminal ends of the representative peptide LfcinB(20-25), which is known as the LfcinB antimicrobial core. All of the eight LfcinB-derived peptides showed in vitro inhibitory effects on ACE activity with different IC(50) values. Moreover, seven of them showed ex vivo inhibitory effects on ACE-dependent vasoconstriction. No clear correlation between in vitro and ex vivo inhibitory effects was found. Only LfcinB(20-25) and one of its fragments, F1, generated after a simulated gastrointestinal digestion, showed significant antihypertensive effects in SHR after oral administration. Remarkably, F1 did not show any effect on ACE-dependent vasoconstriction in contrast to the inhibitory effect showed by LfcinB(20-25). In conclusion, two LfcinB-derived peptides lower blood pressure and exhibit potential as orally effective antihypertensive compounds, yet a complete elucidation of the mechanism(s) involved deserves further ongoing research.

  4. Pulse energy dependence of subcellular dissection by femtosecond laser pulses

    NASA Technical Reports Server (NTRS)

    Heisterkamp, A.; Maxwell, I. Z.; Mazur, E.; Underwood, J. M.; Nickerson, J. A.; Kumar, S.; Ingber, D. E.

    2005-01-01

    Precise dissection of cells with ultrashort laser pulses requires a clear understanding of how the onset and extent of ablation (i.e., the removal of material) depends on pulse energy. We carried out a systematic study of the energy dependence of the plasma-mediated ablation of fluorescently-labeled subcellular structures in the cytoskeleton and nuclei of fixed endothelial cells using femtosecond, near-infrared laser pulses focused through a high-numerical aperture objective lens (1.4 NA). We find that the energy threshold for photobleaching lies between 0.9 and 1.7 nJ. By comparing the changes in fluorescence with the actual material loss determined by electron microscopy, we find that the threshold for true material ablation is about 20% higher than the photobleaching threshold. This information makes it possible to use the fluorescence to determine the onset of true material ablation without resorting to electron microscopy. We confirm the precision of this technique by severing a single microtubule without disrupting the neighboring microtubules, less than 1 micrometer away. c2005 Optical Society of America.

  5. Solution-based single molecule imaging of surface-immobilized conjugated polymers.

    PubMed

    Dalgarno, Paul A; Traina, Christopher A; Penedo, J Carlos; Bazan, Guillermo C; Samuel, Ifor D W

    2013-05-15

    The photophysical behavior of conjugated polymers used in modern optoelectronic devices is strongly influenced by their structural dynamics and conformational heterogeneity, both of which are dependent on solvent properties. Single molecule studies of these polymer systems embedded in a host matrix have proven to be very powerful to investigate the fundamental fluorescent properties. However, such studies lack the possibility of examining the relationship between conformational dynamics and photophysical response in solution, which is the phase from which films for devices are deposited. By developing a synthetic strategy to incorporate a biotin moiety as a surface attachment point at one end of a polyalkylthiophene, we immobilize it, enabling us to make the first single molecule fluorescence measurements of conjugated polymers for long periods of time in solution. We identify fluctuation patterns in the fluorescence signal that can be rationalized in terms of photobleaching and stochastic transitions to reversible dark states. Moreover, by using the advantages of solution-based imaging, we demonstrate that the addition of oxygen scavengers improves optical stability by significantly decreasing the photobleaching rates.

  6. Fabrication of three-dimensional polymer quadratic nonlinear grating structures by layer-by-layer direct laser writing technique

    NASA Astrophysics Data System (ADS)

    Bich Do, Danh; Lin, Jian Hung; Diep Lai, Ngoc; Kan, Hung-Chih; Hsu, Chia Chen

    2011-08-01

    We demonstrate the fabrication of a three-dimensional (3D) polymer quadratic nonlinear (χ(2)) grating structure. By performing layer-by-layer direct laser writing (DLW) and spin-coating approaches, desired photobleached grating patterns were embedded in the guest--host dispersed-red-1/poly(methylmethacrylate) (DR1/PMMA) active layers of an active-passive alternative multilayer structure through photobleaching of DR1 molecules. Polyvinyl-alcohol and SU8 thin films were deposited between DR1/PMMA layers serving as a passive layer to separate DR1/PMMA active layers. After applying the corona electric field poling to the multilayer structure, nonbleached DR1 molecules in the active layers formed polar distribution, and a 3D χ(2) grating structure was obtained. The χ(2) grating structures at different DR1/PMMA nonlinear layers were mapped by laser scanning second harmonic (SH) microscopy, and no cross talk was observed between SH images obtained from neighboring nonlinear layers. The layer-by-layer DLW technique is favorable to fabricating hierarchical 3D polymer nonlinear structures for optoelectronic applications with flexible structural design.

  7. Fabrication of three-dimensional polymer quadratic nonlinear grating structures by layer-by-layer direct laser writing technique.

    PubMed

    Do, Danh Bich; Lin, Jian Hung; Lai, Ngoc Diep; Kan, Hung-Chih; Hsu, Chia Chen

    2011-08-10

    We demonstrate the fabrication of a three-dimensional (3D) polymer quadratic nonlinear (χ(2)) grating structure. By performing layer-by-layer direct laser writing (DLW) and spin-coating approaches, desired photobleached grating patterns were embedded in the guest-host dispersed-red-1/poly(methylmethacrylate) (DR1/PMMA) active layers of an active-passive alternative multilayer structure through photobleaching of DR1 molecules. Polyvinyl-alcohol and SU8 thin films were deposited between DR1/PMMA layers serving as a passive layer to separate DR1/PMMA active layers. After applying the corona electric field poling to the multilayer structure, nonbleached DR1 molecules in the active layers formed polar distribution, and a 3D χ(2) grating structure was obtained. The χ(2) grating structures at different DR1/PMMA nonlinear layers were mapped by laser scanning second harmonic (SH) microscopy, and no cross talk was observed between SH images obtained from neighboring nonlinear layers. The layer-by-layer DLW technique is favorable to fabricating hierarchical 3D polymer nonlinear structures for optoelectronic applications with flexible structural design.

  8. Application of gold nanoparticles as contrast agents in confocal laser scanning microscopy

    NASA Astrophysics Data System (ADS)

    Lemelle, A.; Veksler, B.; Kozhevnikov, I. S.; Akchurin, G. G.; Piletsky, S. A.; Meglinski, I.

    2009-01-01

    Confocal laser scanning microscopy (CLSM) is a modern high-resolution optical technique providing detailed image of tissue structure with high (down to microns) spatial resolution. Aiming at a concurrent improvement of imaging depth and image quality the CLSM requires the use of contrast agents. Commonly employed fluorescent contrast agents, such as fluorescent dyes and proteins, suffer from toxicity, photo-bleaching and overlapping with the tissues autofluorescence. Gold nanoparticles are potentially highly attractive to be applied as a contrast agent since they are not subject to photo-bleaching and can target biochemical cells markers associated with the specific diseases. In current report we consider the applicability of gold nano-spheres as a contrast agent to enhance quality of CLSM images of skin tissues in vitro versus the application of optical clearing agent, such as glycerol. The enhancement of CLSM image contrast was observed with an application of gold nano-spheres diffused within the skin tissues. We show that optical clearing agents such as a glycerol provide better CLSM image contrast than gold nano-spheres.

  9. Biocompatible fluorescent silicon nanocrystals for single-molecule tracking and fluorescence imaging

    PubMed Central

    Nishimura, Hirohito; Ritchie, Ken; Kasai, Rinshi S.; Goto, Miki; Morone, Nobuhiro; Sugimura, Hiroyuki; Tanaka, Koichiro; Sase, Ichiro; Yoshimura, Akihiko; Nakano, Yoshitaro; Fujiwara, Takahiro K.

    2013-01-01

    Fluorescence microscopy is used extensively in cell-biological and biomedical research, but it is often plagued by three major problems with the presently available fluorescent probes: photobleaching, blinking, and large size. We have addressed these problems, with special attention to single-molecule imaging, by developing biocompatible, red-emitting silicon nanocrystals (SiNCs) with a 4.1-nm hydrodynamic diameter. Methods for producing SiNCs by simple chemical etching, for hydrophilically coating them, and for conjugating them to biomolecules precisely at a 1:1 ratio have been developed. Single SiNCs neither blinked nor photobleached during a 300-min overall period observed at video rate. Single receptor molecules in the plasma membrane of living cells (using transferrin receptor) were imaged for ≥10 times longer than with other probes, making it possible for the first time to observe the internalization process of receptor molecules at the single-molecule level. Spatial variations of molecular diffusivity in the scale of 1–2 µm, i.e., a higher level of domain mosaicism in the plasma membrane, were revealed. PMID:24043702

  10. Development of an accurate and high-throughput methodology for structural comprehension of chlorophylls derivatives. (I) Phytylated derivatives.

    PubMed

    Chen, Kewei; Ríos, José Julián; Pérez-Gálvez, Antonio; Roca, María

    2015-08-07

    Phytylated chlorophyll derivatives undergo specific oxidative reactions through the natural metabolism or during food processing or storage, and consequently pyro-, 13(2)-hydroxy-, 15(1)-hydroxy-lactone chlorophylls, and pheophytins (a and b) are originated. New analytical procedures have been developed here to reproduce controlled oxidation reactions that specifically, and in reasonable amounts, produce those natural target standards. At the same time and under the same conditions, 16 natural chlorophyll derivatives have been analyzed by APCI-HPLC-hrMS(2) and most of them by the first time. The combination of the high-resolution MS mode with powerful post-processing software has allowed the identification of new fragmentation patterns, characterizing specific product ions for some particular standards. In addition, new hypotheses and reaction mechanisms for the established MS(2)-based reactions have been proposed. As a general rule, the main product ions involve the phytyl and the propionic chains but the introduction of oxygenated functional groups at the isocyclic ring produces new and specific productions and at the same time inhibits some particular fragmentations. It is noteworthy that all b derivatives, except 15(1)-hydroxy-lactone compounds, undergo specific CO losses. We propose a new reaction mechanism based in the structural configuration of a and b chlorophyll derivatives that explain the exclusive CO fragmentation in all b series except for 15(1)-hydroxy-lactone b and all a series compounds. Copyright © 2015 Elsevier B.V. All rights reserved.

  11. General hybrid projective complete dislocated synchronization with non-derivative and derivative coupling based on parameter identification in several chaotic and hyperchaotic systems

    NASA Astrophysics Data System (ADS)

    Sun, Jun-Wei; Shen, Yi; Zhang, Guo-Dong; Wang, Yan-Feng; Cui, Guang-Zhao

    2013-04-01

    According to the Lyapunov stability theorem, a new general hybrid projective complete dislocated synchronization scheme with non-derivative and derivative coupling based on parameter identification is proposed under the framework of drive-response systems. Every state variable of the response system equals the summation of the hybrid drive systems in the previous hybrid synchronization. However, every state variable of the drive system equals the summation of the hybrid response systems while evolving with time in our method. Complete synchronization, hybrid dislocated synchronization, projective synchronization, non-derivative and derivative coupling, and parameter identification are included as its special item. The Lorenz chaotic system, Rössler chaotic system, memristor chaotic oscillator system, and hyperchaotic Lü system are discussed to show the effectiveness of the proposed methods.

  12. Selective epitaxial growth of zinc blende-derivative on wurtzite-derivative: the case of polytypic Cu2CdSn(S1-xSex)4 nanocrystals

    NASA Astrophysics Data System (ADS)

    Wu, Liang; Fan, Feng-Jia; Gong, Ming; Ge, Jin; Yu, Shu-Hong

    2014-02-01

    Polytypic nanocrystals with zinc blende (ZB) cores and wurtzite (WZ) arms, such as tetrapod and octopod nanocrystals, have been widely reported. However, polytypic nanocrystals with WZ cores and ZB arms or ends have been rarely reported. Here, we report a facile, solution-based approach to the synthesis of polytypic Cu2CdSn(S1-xSex)4 (CCTSSe) nanocrystals with ZB-derivative selectively engineered on (000+/-2)WZ facets of WZ-derived cores. Accordingly, two typical morphologies, i.e., bullet-like nanocrystals with a WZ-derivative core and one ZB-derivative end, and rugby ball-like nanocrystals with a WZ-derivative core and two ZB-derivative ends, can be selectively prepared. The epitaxial growth mechanism is confirmed by the time-dependent experiments. The ratio of rugby ball-like and bullet-like polytypic CCTSSe nanocrystals can be tuned through changing the amount of Cd precursor to adjust the reactivity difference between (0002)WZ and (000-2)WZ facets. These unique polytypic CCTSSe nanocrystals may find applications in energetic semiconducting materials for energy conversion in the future.Polytypic nanocrystals with zinc blende (ZB) cores and wurtzite (WZ) arms, such as tetrapod and octopod nanocrystals, have been widely reported. However, polytypic nanocrystals with WZ cores and ZB arms or ends have been rarely reported. Here, we report a facile, solution-based approach to the synthesis of polytypic Cu2CdSn(S1-xSex)4 (CCTSSe) nanocrystals with ZB-derivative selectively engineered on (000+/-2)WZ facets of WZ-derived cores. Accordingly, two typical morphologies, i.e., bullet-like nanocrystals with a WZ-derivative core and one ZB-derivative end, and rugby ball-like nanocrystals with a WZ-derivative core and two ZB-derivative ends, can be selectively prepared. The epitaxial growth mechanism is confirmed by the time-dependent experiments. The ratio of rugby ball-like and bullet-like polytypic CCTSSe nanocrystals can be tuned through changing the amount of Cd precursor

  13. Immunogenicity of transgenic plant-derived hepatitis B surface antigen.

    PubMed Central

    Thanavala, Y; Yang, Y F; Lyons, P; Mason, H S; Arntzen, C

    1995-01-01

    The focus of the Children's Vaccine Initiative is to encourage the discovery of technology that will make vaccines more readily available to developing countries. Our strategy has been to genetically engineer plants so that they can be used as inexpensive alternatives to fermentation systems for production of subunit antigens. In this paper we report on the immunological response elicited in vivo by using recombinant hepatitis B surface antigen (rHBsAg) purified from transgenic tobacco leaves. The anti-hepatitis B response to the tobacco-derived rHBsAg was qualitatively similar to that obtained by immunizing mice with yeast-derived rHBsAg (commercial vaccine). Additionally, T cells obtained from mice primed with the tobacco-derived rHBsAg could be stimulated in vitro by the tobacco-derived rHBsAg, yeast-derived rHBsAg, and by a synthetic peptide that represents part of the a determinant located in the S region (139-147) of HBsAg. Further support for the integrity of the T-cell epitope of the tobacco-derived rHBsAg was obtained by testing the ability of the primed T cells to proliferate in vitro after stimulation with a monoclonal anti-idiotype and an anti-idiotype-derived peptide, both of which mimic the group-specific a determinant of HBsAg. In total, we have conclusively demonstrated that both B- and T-cell epitopes of HBsAg are preserved when the antigen is expressed in a transgenic plant. PMID:7724566

  14. Animal derived products may conflict with religious patients' beliefs.

    PubMed

    Eriksson, Axelina; Burcharth, Jakob; Rosenberg, Jacob

    2013-12-01

    Implants and drugs with animal and human derived content are widely used in medicine and surgery, but information regarding ingredients is rarely obtainable by health practitioners. A religious perspective concerning the use of animal and human derived drug ingredients has not thoroughly been investigated. The purpose of this study was to clarify which parts of the medical and surgical treatments offered in western world-hospitals that conflicts with believers of major religions. Religious and spiritual leaders of the six largest religions worldwide (18 branches) were contacted. A standardised questionnaire was sent out regarding their position on the use of human and animal derived products in medical and surgical treatments. Of the 18 contacted religious branches, 10 replied representing the 6 largest religions worldwide. Hindus and Sikhs did not approve of the use of bovine or porcine derived products, and Muslims did not accept the use of porcine derived drugs, dressings or implants. Christians (including Jehovah's Witnesses), Jews and Buddhists accepted the use of all animal and human derived products. However, all religions accepted the use of all these products in case of an emergency and only if alternatives were not available. The views here suggest that religious codes conflict with some treatment regimens. It is crucial to obtain informed consent from patients for the use of drugs and implants with animal or human derived content. However, information on the origin of ingredients in drugs is not always available to health practitioners.

  15. [Stereoselective synthesis of polyhydroxylated amines using (S)-pyroglutamic acid derivatives].

    PubMed

    Ikota, Nobuo

    2014-01-01

    Naturally occurring polyhydroxylated amines such as (+)-1-deoxynojirimycin, polyoxamic acid, anisomycin, (-)swainsonine, and alexine stereoisomers, which have interesting biological activities including glucosidase- and mannosidase-inhibitory activity, immunoregulatory activity, and antibacterial effects, were synthesized stereoselectively starting from (S)-pyroglutamic acid derivatives. α,β-Unsaturated lactams ((S)-5-hydroxymethyl-2-oxo-3-pyrroline derivatives), α,β-unsaturated δ-lactone ((S)-4-amino-2-penten-5-olide derivative), and E-olefin ((S,E)-methyl-4-amino-5-hydroxypent-2-enoate derivative) from (S)-pyroglutamic acid derivatives were dihydroxylated using OsO4 in the presence of N-methyl morpholine N-oxide (NMO) to afford various chiral building blocks with different configurations. The stereoselectivity of cis-dihydroxylation for α,β-unsaturated lactams and α,β-unsaturated δ-lactone was very high, while the stereoselectivity was low for E-olefin. Therefore, the double asymmetric induction of E-olefin using K2OsO4 with chiral ligands was successively applied to yield high stereoselectivity. (2R,3S)-2-Hydroxymethyl-3-hydroxypyrrolidine and Gaissman-Weiss lactone, important intermediates for the preparation of pyrrolizidine alkaloids, were synthesized from a (3R,4R,5R)-3,4-dihydroxy-5-hydroxymethyl-2-pyrrolidinone derivative derived from α,β-unsatulated lactam. (+)-1-Deoxynojirimycin was synthesized from a (2S,3R,4R)-methyl 4-amino-2,3,5-trihydroxypentanoate derivative of E-olefin. (-)-Swainsonine and its stereoisomers were synthesized from (2R,3S,4R)- or (2R,3R,4R)-2-hydroxymethyl-3,4-dihydroxypyrrolidine derivatives of α,β-unsaturated δ-lactone or α,β-unsaturated lactam. The key reaction was diastereoselective allylation of the aldehyde derived from the corresponding 2-hydroxymethylpyrrolidine derivatives with various allylation reagents. The high diastereoselectivity could be explained by cyclic chelate formation between metals and the

  16. Kinks in higher derivative scalar field theory

    NASA Astrophysics Data System (ADS)

    Zhong, Yuan; Guo, Rong-Zhen; Fu, Chun-E.; Liu, Yu-Xiao

    2018-07-01

    We study static kink configurations in a type of two-dimensional higher derivative scalar field theory whose Lagrangian contains second-order derivative terms of the field. The linear fluctuation around arbitrary static kink solutions is analyzed. We find that, the linear spectrum can be described by a supersymmetric quantum mechanics problem, and the criteria for stable static solutions can be given analytically. We also construct a superpotential formalism for finding analytical static kink solutions. Using this formalism we first reproduce some existed solutions and then offer a new solution. The properties of our solution is studied and compared with those preexisted. We also show the possibility in constructing twinlike model in the higher derivative theory, and give the consistency conditions for twinlike models corresponding to the canonical scalar field theory.

  17. Second derivatives for approximate spin projection methods

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

    Thompson, Lee M.; Hratchian, Hrant P., E-mail: hhratchian@ucmerced.edu

    2015-02-07

    The use of broken-symmetry electronic structure methods is required in order to obtain correct behavior of electronically strained open-shell systems, such as transition states, biradicals, and transition metals. This approach often has issues with spin contamination, which can lead to significant errors in predicted energies, geometries, and properties. Approximate projection schemes are able to correct for spin contamination and can often yield improved results. To fully make use of these methods and to carry out exploration of the potential energy surface, it is desirable to develop an efficient second energy derivative theory. In this paper, we formulate the analytical secondmore » derivatives for the Yamaguchi approximate projection scheme, building on recent work that has yielded an efficient implementation of the analytical first derivatives.« less

  18. Polymer-Derived Boron Nitride: A Review on the Chemistry, Shaping and Ceramic Conversion of Borazine Derivatives

    PubMed Central

    Bernard, Samuel; Miele, Philippe

    2014-01-01

    Boron nitride (BN) is a III-V compound which is the focus of important research since its discovery in the early 19th century. BN is electronic to carbon and thus, in the same way that carbon exists as graphite, BN exists in the hexagonal phase. The latter offers an unusual combination of properties that cannot be found in any other ceramics. However, these properties closely depend on the synthesis processes. This review states the recent developments in the preparation of BN through the chemistry, shaping and ceramic conversion of borazine derivatives. This concept denoted as Polymer-Derived Ceramics (PDCs) route allows tailoring the chemistry of precursors to elaborate complex BN shapes which cannot be obtained by conventional process. The effect of the chemistry of the molecular precursors, i.e., borazine and trichloroborazine, and their polymeric derivatives i.e., polyborazylene and poly[tri(methylamino)borazine], in which the specific functional groups and structural motifs determine the shaping potential by conventional liquid-phase process and plastic-forming techniques is discussed. Nanotubes, nano-fibers, coatings, monoliths and fiber-reinforced matrix composites are especially described. This leads to materials which are of significant engineering interest. PMID:28788257

  19. Polymer-Derived Boron Nitride: A Review on the Chemistry, Shaping and Ceramic Conversion of Borazine Derivatives.

    PubMed

    Bernard, Samuel; Miele, Philippe

    2014-11-21

    Boron nitride (BN) is a III-V compound which is the focus of important research since its discovery in the early 19th century. BN is electronic to carbon and thus, in the same way that carbon exists as graphite, BN exists in the hexagonal phase. The latter offers an unusual combination of properties that cannot be found in any other ceramics. However, these properties closely depend on the synthesis processes. This review states the recent developments in the preparation of BN through the chemistry, shaping and ceramic conversion of borazine derivatives. This concept denoted as Polymer-Derived Ceramics (PDCs) route allows tailoring the chemistry of precursors to elaborate complex BN shapes which cannot be obtained by conventional process. The effect of the chemistry of the molecular precursors, i.e. , borazine and trichloroborazine, and their polymeric derivatives i.e. , polyborazylene and poly[tri(methylamino)borazine], in which the specific functional groups and structural motifs determine the shaping potential by conventional liquid-phase process and plastic-forming techniques is discussed. Nanotubes, nano-fibers, coatings, monoliths and fiber-reinforced matrix composites are especially described. This leads to materials which are of significant engineering interest.

  20. On the evaluation of derivatives of Gaussian integrals

    NASA Technical Reports Server (NTRS)

    Helgaker, Trygve; Taylor, Peter R.

    1992-01-01

    We show that by a suitable change of variables, the derivatives of molecular integrals over Gaussian-type functions required for analytic energy derivatives can be evaluated with significantly less computational effort than current formulations. The reduction in effort increases with the order of differentiation.

  1. Healthy degenerate theories with higher derivatives

    NASA Astrophysics Data System (ADS)

    Motohashi, Hayato; Noui, Karim; Suyama, Teruaki; Yamaguchi, Masahide; Langlois, David

    2016-07-01

    In the context of classical mechanics, we study the conditions under which higher-order derivative theories can evade the so-called Ostrogradsky instability. More precisely, we consider general Lagrangians with second order time derivatives, of the form L(̈phia, dot phia, phia; qi, qi) with a = 1,⋯,n and i = 1,⋯,m. For n = 1, assuming that the qi's form a nondegenerate subsystem, we confirm that the degeneracy of the kinetic matrix eliminates the Ostrogradsky instability. The degeneracy implies, in the Hamiltonian formulation of the theory, the existence of a primary constraint, which generates a secondary constraint, thus eliminating the Ostrogradsky ghost. For n > 1, we show that, in addition to the degeneracy of the kinetic matrix, one needs to impose extra conditions to ensure the presence of a sufficient number of secondary constraints that can eliminate all the Ostrogradsky ghosts. When these conditions that ensure the disappearance of the Ostrogradsky instability are satisfied, we show that the Euler-Lagrange equations, which involve a priori higher order derivatives, can be reduced to a second order system.

  2. Improving ready biodegradability testing of fatty amine derivatives.

    PubMed

    van Ginkel, C G; Gancet, C; Hirschen, M; Galobardes, M; Lemaire, Ph; Rosenblom, J

    2008-09-01

    This study assesses the biodegradation potential of a number of fatty amine derivatives in tests following the OECD guidelines for ready biodegradability. A number of methods are used to reduce toxicity and improve the bioavailability of the fatty amine derivatives in these tests. Alkyl-1,3-diaminopropanes and octadecyltrimethylammonium chloride are toxic to microorganisms at concentrations used in OECD ready biodegradability tests. The concentration of these fatty amine derivatives in the aqueous phase can be reduced by reacting humic, or lignosulphonic acids with the derivatives or through the addition of silica gel to the test bottles. Using these non-biodegradable substances, ready biodegradability test results were obtained with tallow-1,3-diaminopropane and octadecyltrimethylammonium chloride. Demonstration of the ready biodegradability of the water-insoluble dioctadecylamine under the prescribed standard conditions is almost impossible due to the limited bioavailability of this compound. However, ready biodegradability results were achieved by using very low initial test substance concentrations and by introducing an organic phase. The contents of the bottles used to assess the biodegradability of dioctadecylamine were always mixed. False negative biodegradability results obtained with the fatty amine derivatives studied are the result of toxic effects and/or limited bioavailability. The aids investigated therefore improve ready biodegradability testing.

  3. 3-Coumaranone derivatives as inhibitors of monoamine oxidase.

    PubMed

    Van Dyk, Adriaan S; Petzer, Jacobus P; Petzer, Anél; Legoabe, Lesetja J

    2015-01-01

    The present study examines the monoamine oxidase (MAO) inhibitory properties of a series of 20 3-coumaranone [benzofuran-3(2H)-one] derivatives. The 3-coumaranone derivatives are structurally related to series of α-tetralone and 1-indanone derivatives, which have recently been shown to potently inhibit MAO, with selectivity for MAO-B (in preference to the MAO-A isoform). 3-Coumaranones are similarly found to selectively inhibit human MAO-B with half-maximal inhibitory concentration (IC50) values of 0.004-1.05 µM. Nine compounds exhibited IC50<0.05 µM for the inhibition of MAO-B. For the inhibition of human MAO-A, IC50 values ranged from 0.586 to >100 µM, with only one compound possessing an IC50<1 µM. For selected 3-coumaranone derivatives, it is established that MAO-A and MAO-B inhibition are reversible since dialysis of enzyme-inhibitor mixtures almost completely restores enzyme activity. On the basis of the selectivity profiles and potent action, it may be concluded that the 3-coumaranone derivatives are suitable leads for the development of selective MAO-B inhibitors as potential treatment for disorders such as Parkinson's disease and Alzheimer's disease.

  4. 3-Coumaranone derivatives as inhibitors of monoamine oxidase

    PubMed Central

    Van Dyk, Adriaan S; Petzer, Jacobus P; Petzer, Anél; Legoabe, Lesetja J

    2015-01-01

    The present study examines the monoamine oxidase (MAO) inhibitory properties of a series of 20 3-coumaranone [benzofuran-3(2H)-one] derivatives. The 3-coumaranone derivatives are structurally related to series of α-tetralone and 1-indanone derivatives, which have recently been shown to potently inhibit MAO, with selectivity for MAO-B (in preference to the MAO-A isoform). 3-Coumaranones are similarly found to selectively inhibit human MAO-B with half-maximal inhibitory concentration (IC50) values of 0.004–1.05 µM. Nine compounds exhibited IC50<0.05 µM for the inhibition of MAO-B. For the inhibition of human MAO-A, IC50 values ranged from 0.586 to >100 µM, with only one compound possessing an IC50<1 µM. For selected 3-coumaranone derivatives, it is established that MAO-A and MAO-B inhibition are reversible since dialysis of enzyme–inhibitor mixtures almost completely restores enzyme activity. On the basis of the selectivity profiles and potent action, it may be concluded that the 3-coumaranone derivatives are suitable leads for the development of selective MAO-B inhibitors as potential treatment for disorders such as Parkinson’s disease and Alzheimer’s disease. PMID:26491258

  5. Benzoxazole derivatives suppress lipopolysaccharide-induced mast cell activation.

    PubMed

    Cho, Kyung-Ah; Park, Minhwa; Kim, Yu-Hee; Choo, Hea-Young Park; Lee, Kyung Ho

    2018-05-01

    Mast cells are central regulators of allergic inflammation that function by releasing various proallergic inflammatory mediators, including histamine, eicosanoids and proinflammatory cytokines. Occasionally, bacterial infections may initiate or worsen allergic inflammation. A number of studies have indicated that activation of lipoxygenase in mast cells positive regulates allergic inflammatory responses by generating leukotrienes and proinflammatory cytokines. In the present study, the effects of benzoxazole derivatives on the lipopolysaccharide (LPS)‑induced expression of proinflammatory cytokines, production of histamine and surface expression of co‑stimulatory molecules on bone marrow-derived mast cells (BMMCs) were studied. The benzoxazole derivatives significantly reduced the expression of interleukin (IL)‑1β, IL‑6, IL‑13, tumor necrosis factor‑α, perilipin (PLIN) 2, and PLIN3 in BMMCs treated with LPS. Furthermore, histamine production was suppressed in BMMCs treated with LPS, or treated with phorbol-12-myristate-13-acetate/ionomycin. Benzoxazole derivatives marginally affected the surface expression of cluster of differentiation (CD)80 and CD86 on BMMCs in the presence of LPS, although LPS alone did not increase the expression of those proteins. Therefore, benzoxazole derivatives inhibited the secretion of proinflammatory cytokines in mast cells and may be potential candidate anti‑allergic agents to suppress mast cell activation.

  6. 40 CFR 721.4520 - Isopropylidene, bis(1,1-dimethylpropyl) derivative.

    Code of Federal Regulations, 2010 CFR

    2010-07-01

    ...) derivative. 721.4520 Section 721.4520 Protection of Environment ENVIRONMENTAL PROTECTION AGENCY (CONTINUED... Specific Chemical Substances § 721.4520 Isopropylidene, bis(1,1-dimethylpropyl) derivative. (a) Chemical... isopropylidene, bis(1,1-dimethylpropyl) derivative (PMN P-85-648) is subject to reporting under this section for...

  7. 40 CFR 721.4520 - Isopropylidene, bis(1,1-dimethylpropyl) derivative.

    Code of Federal Regulations, 2011 CFR

    2011-07-01

    ...) derivative. 721.4520 Section 721.4520 Protection of Environment ENVIRONMENTAL PROTECTION AGENCY (CONTINUED... Specific Chemical Substances § 721.4520 Isopropylidene, bis(1,1-dimethylpropyl) derivative. (a) Chemical... isopropylidene, bis(1,1-dimethylpropyl) derivative (PMN P-85-648) is subject to reporting under this section for...

  8. Extraction of stability and control derivatives from orbiter flight data

    NASA Technical Reports Server (NTRS)

    Iliff, Kenneth W.; Shafer, Mary F.

    1993-01-01

    The Space Shuttle Orbiter has provided unique and important information on aircraft flight dynamics. This information has provided the opportunity to assess the flight-derived stability and control derivatives for maneuvering flight in the hypersonic regime. In the case of the Space Shuttle Orbiter, these derivatives are required to determine if certain configuration placards (limitations on the flight envelope) can be modified. These placards were determined on the basis of preflight predictions and the associated uncertainties. As flight-determined derivatives are obtained, the placards are reassessed, and some of them are removed or modified. Extraction of the stability and control derivatives was justified by operational considerations and not by research considerations. Using flight results to update the predicted database of the orbiter is one of the most completely documented processes for a flight vehicle. This process followed from the requirement for analysis of flight data for control system updates and for expansion of the operational flight envelope. These results show significant changes in many important stability and control derivatives from the preflight database. This paper presents some of the stability and control derivative results obtained from Space Shuttle flights. Some of the limitations of this information are also examined.

  9. Improving your four-dimensional image: traveling through a decade of light-sheet-based fluorescence microscopy research.

    PubMed

    Strobl, Frederic; Schmitz, Alexander; Stelzer, Ernst H K

    2017-06-01

    Light-sheet-based fluorescence microscopy features optical sectioning in the excitation process. This reduces phototoxicity and photobleaching by up to four orders of magnitude compared with that caused by confocal fluorescence microscopy, simplifies segmentation and quantification for three-dimensional cell biology, and supports the transition from on-demand to systematic data acquisition in developmental biology applications.

  10. Special relativity derived from spacetime magma.

    PubMed

    Greensite, Fred

    2014-01-01

    We present a derivation of relativistic spacetime largely untethered from specific physical considerations, in constrast to the many physically-based derivations that have appeared in the last few decades. The argument proceeds from the inherent magma (groupoid) existing on the union of spacetime frame components [Formula: see text] and Euclidean [Formula: see text] which is consistent with an "inversion symmetry" constraint from which the Minkowski norm results. In this context, the latter is also characterized as one member of a class of "inverse norms" which play major roles with respect to various unital [Formula: see text]-algebras more generally.

  11. Gauge fixing in higher-derivative gravity

    NASA Astrophysics Data System (ADS)

    Bartoli, A.; Julve, J.; Sánchez, E. J.

    1999-07-01

    Linearized 4-derivative gravity with a general gauge-fixing term is considered. By a Legendre transform and a suitable diagonalization procedure it is cast into a second-order equivalent form where the nature of the physical degrees of freedom, the gauge ghosts, the Weyl ghosts and the intriguing `third ghosts', characteristic to higher-derivative theories, is made explicit. The symmetries of the theory and the structure of the compensating Faddeev-Popov ghost sector exhibit non-trivial peculiarities. The unitarity breaking negative-norm Weyl ghosts, already present in the diff-invariant theory, are out of the reach of the ghost cancellation BRST mechanism.

  12. Animal derived products may conflict with religious patients’ beliefs

    PubMed Central

    2013-01-01

    Background Implants and drugs with animal and human derived content are widely used in medicine and surgery, but information regarding ingredients is rarely obtainable by health practitioners. A religious perspective concerning the use of animal and human derived drug ingredients has not thoroughly been investigated. The purpose of this study was to clarify which parts of the medical and surgical treatments offered in western world-hospitals that conflicts with believers of major religions. Methods Religious and spiritual leaders of the six largest religions worldwide (18 branches) were contacted. A standardised questionnaire was sent out regarding their position on the use of human and animal derived products in medical and surgical treatments. Results Of the 18 contacted religious branches, 10 replied representing the 6 largest religions worldwide. Hindus and Sikhs did not approve of the use of bovine or porcine derived products, and Muslims did not accept the use of porcine derived drugs, dressings or implants. Christians (including Jehovah’s Witnesses), Jews and Buddhists accepted the use of all animal and human derived products. However, all religions accepted the use of all these products in case of an emergency and only if alternatives were not available. Conclusions The views here suggest that religious codes conflict with some treatment regimens. It is crucial to obtain informed consent from patients for the use of drugs and implants with animal or human derived content. However, information on the origin of ingredients in drugs is not always available to health practitioners. PMID:24289542

  13. Adipose-derived cells.

    PubMed

    Meliga, Emanuele; Strem, Brian M; Duckers, H J; Serruys, Patrick W

    2007-01-01

    Heart failure is by far the most common cause of hospitalization in Western countries, with onerous economic consequences. Cell therapy holds great promise for use in tissue regeneration and is increasingly used in an effort to improve outcomes in cardiac disease. Recently it has been shown that adipose tissue, in addition to committed adipogenic, endothelial progenitor cells and pluripotent vascular progenitor cells, also contains multipotent cell types (adipose-derived stem cells, ADSCs) that, in cell culture conditions, have shown to have an impressive developmental plasticity including the ability to undergo multilineage differentiation and self-renewal. ADSCs express multiple CD marker antigens similar to those observed on MSCs and are also capable of secreting a large number of angiogenesis-related cytokines, including vascular endothelial growth factor, granulocyte/macrophage colony stimulating factor, stromal-derived factor-1alpha, and hepatocyte growth factor. Adipose tissue can be harvested in large quantities with minimal morbidity in several regions of the body and, on average, 100 ml of human adipose tissue yields about 1 x 10(6) stem cells. Studies conducted in porcine AMI models have shown a significant LV functional improvement, with no report of any potentially fatal arrhythmias. The APOLLO trial, a prospective, double blind, randomized, placebo-controlled trial currently in the recruiting phase, is a "first-in-man" study that explores the safety and feasibility of ADSC transplantation in patients with acute MI.

  14. 19 CFR 191.173 - Imported duty-paid derivatives (no manufacture).

    Code of Federal Regulations, 2011 CFR

    2011-04-01

    ... 19 Customs Duties 2 2011-04-01 2011-04-01 false Imported duty-paid derivatives (no manufacture... SECURITY; DEPARTMENT OF THE TREASURY (CONTINUED) DRAWBACK Substitution of Finished Petroleum Derivatives § 191.173 Imported duty-paid derivatives (no manufacture). When the basis for drawback under 19 U.S.C...

  15. 12 CFR 223.33 - What rules apply to derivative transactions?

    Code of Federal Regulations, 2011 CFR

    2011-01-01

    ... 12 Banks and Banking 3 2011-01-01 2011-01-01 false What rules apply to derivative transactions... Requirements Under Section 23A § 223.33 What rules apply to derivative transactions? (a) Market terms requirement. Derivative transactions between a member bank and its affiliates (other than depository...

  16. The Processing of Derivational Morphology in Korean-English Bilingual Readers

    ERIC Educational Resources Information Center

    Kim, Say Young; Wang, Min; Ko, In Yeong

    2011-01-01

    Three experiments using a priming lexical decision paradigm were conducted to examine whether cross-language activation occurs via decomposition during the processing of derived words in Korean-English bilingual readers. In Experiment 1, when participants were given a real derived word and an interpretable derived pseudoword (i.e., illegal…

  17. Enzymatic synthesis of novel quercetin sialyllactoside derivatives.

    PubMed

    Darsandhari, Sumangala; Bae, Jae Yoon; Shrestha, Biplav; Yamaguchi, Tokutaro; Jung, Hye Jin; Han, Jang Mi; Rha, Chan-Su; Pandey, Ramesh Prasad; Sohng, Jae Kyung

    2018-06-06

    Quercetin and its derivatives are important flavonols that show diverse biological activity, such as antioxidant, anticarcinogenic, anti-inflammatory, and antiviral activities. Adding different substituents to quercetin may change the biochemical activity and bioavailability of molecules, when compared to the aglycone. Here, we have synthesised two novel derivatives of quercetin, quercetin-3-O-β-d-glucopyranosyl, 4''-O-d-galactopyranosyl 3'''-O-α-N-acetyl neuraminic acid i.e. 3'-sialyllactosyl quercetin (3'SL-Q) and quercetin-3-O-β-d-glucopyranosyl, 4''-O-β-d-galactopyranosyl 6'''-O-α-N-acetyl neuraminic acid i.e. 6'-sialyllactosyl quercetin (6'SL-Q) with the use of glycosyltransferases and sialyltransferases enzymes. These derivatives of quercetin were characterised by high-resolution quadrupole-time-of-flight electrospray ionisation mass spectrometry (HR-QTOF-ESI/MS) and 1 H and 13 C nuclear magnetic resonance (NMR) analyses.

  18. Derivation of GNSS derived station velocities for a surface deformation model in the Austrian region

    NASA Astrophysics Data System (ADS)

    Umnig, Elke; Weber, Robert; Maras, Jadre; Brückl, Ewald

    2016-04-01

    This contribution deals with the first comprehensive analysis of GNSS derived surface velocities computed within an observation network of about 100 stations covering the whole Austrian territory and parts of the neighbouring countries. Coordinate time series are available now, spanning a period of 5 years (2010.0-2015.0) for one focus area in East Austria and one and a half year (2013.5-2015.0) for the remaining part of the tracking network. In principle the data series are stemming from two different GNSS campaigns. The former was set up to investigate intra plate tectonic movements within the framework of the project ALPAACT (seismological and geodetic monitoring of ALpine-PAnnonian ACtive Tectonics), the latter was designed to support a number of various requests, e.g. derivation of GNSS derived water vapour fields, but also to expand the foresaid tectonic studies. In addition the activities within the ALPAACT project supplement the educational initiative SHOOLS & QUAKES, where scholars contribute to seismological research. For the whole period of the processed coordinate time series daily solutions have been computed by means of the Bernese software. The processed coordinate time series are tied to the global reference frame ITRF2000 as well as to the frame ITRF2008. Due to the transition of the reference from ITRF2000 to ITRF2008 within the processing period, but also due to updates of the Bernese software from version 5.0 to 5.2 the time series were initially not fully consistent and have to be re-aligned to a common frame. So the goal of this investigation is to derive a nationwide consistent horizontal motion field on base of GNSS reference station data within the ITRF2008 frame, but also with respect to the Eurasian plate. In this presentation we focus on the set-up of the coordinate time series and on the problem of frame alignment. Special attention is also paid to the separation into linear and periodic motion signals, originating from tectonic or non

  19. Isocoumarin derivatives from the endophytic fungus, Pestalotiopsis sp.

    PubMed

    Song, Ren-Yu; Wang, Xiao-Bing; Yin, Guo-Ping; Liu, Rui-Huan; Kong, Ling-Yi; Yang, Ming-Hua

    2017-10-01

    Five new isocoumarin derivatives, pestalactone A-C (1-3) and pestapyrone D-E (4-5), together with two known compounds (6-7) were isolated from the solid cultures of the endophytic fungus Pestalotiopsis sp. obtained from Photinia frasery. Their structures were mainly determined by extensive spectroscopic analysis, Mo 2 (OCOCH 3 ) 4 -induced electronic circular dichroism (ECD), and ECD calculation. Compounds 1 and 2 were rare isocoumarin derivatives and derived from distinctive polyketide pathways. Compound 3 exhibited potent antifungal activity against Candida glabrata (ATCC 90030) with an MIC 50 value of 3.49±0.21μg/mL. Copyright © 2017. Published by Elsevier B.V.

  20. Molecular Docking Study on Galantamine Derivatives as Cholinesterase Inhibitors.

    PubMed

    Atanasova, Mariyana; Yordanov, Nikola; Dimitrov, Ivan; Berkov, Strahil; Doytchinova, Irini

    2015-06-01

    A training set of 22 synthetic galantamine derivatives binding to acetylcholinesterase was docked by GOLD and the protocol was optimized in terms of scoring function, rigidity/flexibility of the binding site, presence/absence of a water molecule inside and radius of the binding site. A moderate correlation was found between the affinities of compounds expressed as pIC50 values and their docking scores. The optimized docking protocol was validated by an external test set of 11 natural galantamine derivatives and the correlation coefficient between the docking scores and the pIC50 values was 0.800. The derived relationship was used to analyze the interactions between galantamine derivatives and AChE. © 2015 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.