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Sample records for acridines

  1. Hydrodenitrogenation of quinoline and acridine

    Energy Technology Data Exchange (ETDEWEB)

    Reiff, Jr., E. K.

    1977-06-01

    The hydrodenitrogenation of quinoline and of acridine was studied in a batch autoclave reactor between 342 and 353/sup 0/C and between 500 and 2000 psig. The several commercial hydrotreating catalysts examined decreased in activity in the following order for quinoline hydrodenitrogenation: Ni--Mo/Al/sub 2/O/sub 3/, Ni--W/Al/sub 2/O/sub 3/, Ni--W/SiO/sub 2/--Al/sub 2/O/sub 3/, and Co--Mo/Al/sub 2/O/sub 3/. The total nitrogen removal rate for quinoline was slightly greater than that for acridine and both followed pseudo first-order kinetics over a conversion range of 0 to 50%. Hydrogenation and cracking steps were both kinetically limiting. Nitrogen-containing reaction products for quinoline hydrodenitrogenation were 1,2,3,4-tetrahydroquinoline, 5,6,7,8-tetrahydroquinoline, decahydroquinoline and o-propylaniline. At 342/sup 0/C and 500 psig quinoline and 1,2,3,4-tetrahydroquinoline were in thermodynamic equilibrium, and the disappearance of the lumped group of quinoline plus 1,2,3,4-tetrahydroquinoline followed pseudo first-order kinetics. Sixteen nitrogen-containing reaction products were found for acridine hydrodenitrogenation, including 1,2,3,4-tetrahydroacridine, 1,2,3,4,9,10,13,14-octahydroacridine, sym-octahydroacridine, perhydroacridine, and o-(methylenecyclohexane)aniline. The hydrogenolysis step for both quinoline and acridine appears to be through hydrogenated forms of these compounds. This is supported by bond strength arguments.

  2. Methyltrioctylammonium chloride catalysed sonochemical synthesis of acridine diones

    Indian Academy of Sciences (India)

    Bhupinder Kaur; Harish Kumar

    2013-09-01

    The greener, clean and efficient protocol for the synthesis of acridine diones derivatives has been achieved by reacting aromatic aldehyde, dimedone and amines using methyltrioctylammonium chloride (Aliquate 336) as a catalyst under ultrasonic irradiations.

  3. Antitumor polycyclic acridines. 7. Synthesis and biological properties of DNA affinic tetra- and pentacyclic acridines.

    Science.gov (United States)

    Stanslas, J; Hagan, D J; Ellis, M J; Turner, C; Carmichael, J; Ward, W; Hammonds, T R; Stevens, M F

    2000-04-20

    New synthetic routes to a series of tetra- and pentacyclic acridines related in structure to marine natural products are reported. The novel water-soluble agent dihydroindolizino[7,6,5-kl]acridinium chloride 14 has inhibitory activity in a panel of non-small-cell lung and breast tumor cell lines exceeding that of m-AMSA. The salt inhibited the release of minicircle products of kDNA confirming that disorganization of topoisomerase II partly underlies the activity of the compound. COMPARE analysis of the NCI mean graph profile of compound 14 at the GI(50) level corroborates this conclusion with Pearson correlation coefficients (>0.6) to clinical agents of the topoisomerase II class: however, this correlation was not seen at the LC(50) level. The inhibitory action of 14 on Saccharomyces cerevisiae transfected with human topoisomerase II isoforms showed a 3-fold selectivity against the IIalpha isoform over the IIbeta isoform. Unlike m-AMSA, 14 is not susceptible to P-glycoprotein-mediated drug efflux and retains activity in lung cells with derived resistance to the topoisomerase II inhibitor etoposide.

  4. Photocatalytic Water Splitting with the Acridine Chromophore: A Computational Study.

    Science.gov (United States)

    Liu, Xiaojun; Karsili, Tolga N V; Sobolewski, Andrzej L; Domcke, Wolfgang

    2015-08-20

    The hydrogen-bonded acridine-water complex is considered as a model system for the exploration of photochemical reactions which can lead to the splitting of water into H(•) and OH(•) radicals. The vertical excitation energies of the lowest singlet and triplet excited states of the complex were calculated with the CASSCF/CASPT2 and ADC(2) ab initio electronic-structure methods. In addition to the well-known excited states of the acridine chromophore, excited states of charge-transfer character were identified, in which an electron is transferred from the p orbital of the H2O molecule to the π* orbital of acridine. The low-energy barriers which separate these reactive charge-transfer states from the spectroscopic states of the acridine-water complex have been characterized by the calculation of two-dimensional relaxed potential-energy surfaces as functions of the H atom-transfer coordinate and the donor (O)-acceptor (N) distance. When populated, these charge-transfer states drive the transfer of a proton from the water molecule to acridine, which results in the acridinyl-hydroxyl biradical. The same computational methods were employed to explore the photochemistry of the (N-hydrogenated) acridinyl radical. The latter possesses low-lying (about 3.0 eV) ππ* excited states with appreciable oscillator strengths in addition to a low-lying dark ππ* excited state. The bound potential-energy functions of the ππ* excited states are predissociated by the potential-energy function of an excited state of πσ* character which is repulsive with respect to the NH stretching coordinate. The dissociation threshold of the πσ* state is about 2.7 eV and thus below the excitation energies of the bright ππ* states. The conical intersections of the πσ* state with the ππ* excited states and with the electronic ground state provide a mechanism for the direct and fast photodetachment of the H atom from the acridinyl radical. These computational results indicate that the H2

  5. Optically enhanced nuclear cross polarization in acridine-doped fluorene

    Energy Technology Data Exchange (ETDEWEB)

    Oshiro, C.M.

    1982-06-01

    The objective of this work has been to create large polarizations of the dilute /sup 13/C nuclei in the solid state. The idea was to create /sup 1/H polarizations larger than Boltzmann and to use the proton enhanced nuclear induction spectroscopy cross polarization technique to then transfer this large polarization to the /sup 13/C spin system. Optical Nuclear Polarization (ONP) of acridine-doped fluorene single crystals was studied. In addition, ONP of powdered samples of the acridine-doped fluorene was studied. In general, many compounds do not crystallize easily or do not form large crystals suitable for NMR experiments. Powdered, amorphous and randomly dispersed samples are generally far more readily available than single crystals. One objective of this work has been to (first) create large /sup 1/H polarizations. Although large optical proton polarizations in single crystals have been reported previously, optically generated polarizations in powdered samples have not been reported. For these reasons, ONP studies of powdered samples of the acridine-doped fluorene were also undertaken. Using ONP in combination with the proton enhanced nuclear induction spectroscopy experiment, large /sup 13/C polarizations have been created in fluorene single crystals. These large /sup 13/C polarizations have permitted the determination of the seven incongruent chemical shielding tensors of the fluorene molecule. Part 2 of this thesis describes the proton enhanced nuclear induction spectroscopy experiment. Part 3 describes the ONP experiment. Part 4 is a description of the experimental set-up. Part 5 describes the data analysis for the determination of the chemical shielding tensors. Part 6 presents the results of the ONP experiments performed in this work and the chemical shielding tensors determined.

  6. Rapid diagnosis of bacteremia in adults using acridine orange stained buffy coat smears

    OpenAIRE

    Miller, Mark; Mendelson, Jack

    1990-01-01

    The use of acridine orange stained buffy coat smears was assessed as a rapid screening test for bacteremia in adults. A total of 356 consecutive blood cultures were submitted with simultaneous anticoagulated blood samples, from which a buffy coat smear was prepared and stained with acridine orange (100 mg/L; pH 3.0). Forty-one of 356 blood samples (12%) yielded organisms in the blood culture system. Compared to blood culture, the overall sensitivity of acridine orange stained buffy coat smear...

  7. Interaction of acridine-calix[4]arene with DNA at the electrified liquid liquid interface

    Energy Technology Data Exchange (ETDEWEB)

    Kivlehan, Francine [Tyndall National Institute, Lee Maltings, University College Cork, Cork (Ireland); Lefoix, Myriam; Moynihan, Humphrey A. [Department of Chemistry, Analytical and Biological Chemistry Research Facility, University College Cork, Cork (Ireland); Thompson, Damien; Ogurtsov, Vladimir I.; Herzog, Gregoire [Tyndall National Institute, Lee Maltings, University College Cork, Cork (Ireland); Arrigan, Damien W.M., E-mail: d.arrigan@curtin.edu.a [Tyndall National Institute, Lee Maltings, University College Cork, Cork (Ireland)

    2010-03-30

    The behaviour of an acridine-functionalised calix[4]arene at the interface between two immiscible electrolyte solutions (ITIES) is reported. Molecular modelling showed that the acridine-calix[4]arene has regions of significant net positive charge spread throughout the protonated acridine moieties, consistent with it being able to function as an anion ionophore. The presence of this compound in the organic phase facilitated the transfer of aqueous phase electrolyte ions. Upon addition of double stranded DNA to the aqueous phase, the transfer of electrolyte anions was diminished, due to DNA binding to the acridine moiety at the ITIES. The behaviour provides a basis for DNA hybridization detection using electrochemistry at the ITIES.

  8. Synthesis, DNA Binding, and Antiproliferative Activity of Novel Acridine-Thiosemicarbazone Derivatives

    OpenAIRE

    Sinara Mônica Vitalino de Almeida; Elizabeth Almeida Lafayette; Lúcia Patrícia Bezerra Gomes da Silva; Cézar Augusto da Cruz Amorim; Tiago Bento de Oliveira; Ana Lucia Tasca Gois Ruiz; João Ernesto de Carvalho; Ricardo Olímpio de Moura; Eduardo Isidoro Carneiro Beltrão; Maria do Carmo Alves de Lima; Luiz Bezerra de Carvalho Júnior

    2015-01-01

    In this work, the acridine nucleus was used as a lead-compound for structural modification by adding different substituted thiosemicarbazide moieties. Eight new (Z)-2-(acridin-9-ylmethylene)-N-phenylhydrazinecarbothioamide derivatives (3a–h) were synthesized, their antiproliferative activities were evaluated, and DNA binding properties were performed with calf thymus DNA (ctDNA) by electronic absorption and fluorescence spectroscopies. Both hyperchromic and hypochromic effects, as well as re...

  9. Covalent functionalization of graphene oxide by 9-(4-aminophenyl)acridine and its derivatives

    Institute of Scientific and Technical Information of China (English)

    Yi Si Feng; Jing Jing Ma; Xin Yan Lin; Jia Song Zhang; Peng Lv; Hua Jian Xu; Lin Bao Luo

    2012-01-01

    Graphene/acridine (G-Acr) hybrid structures were synthesized through covalent functionalization of graphene oxide with 9-(4-aminophenyl)acridine (APA) and its derivatives.The G-Acr hybrids were characterized by Fourier transform infrared spectroscopy,ultraviolet-visible spectrophotometry,thermal gravimetric analysis and Raman spectroscopy.X-ray photoelectron spectroscopy confirms that the binding energies of APA and its derivatives shifted to higher values,revealing pronounced charge transfer at the interface of graphene and organic molecules.

  10. A quantitative model for using acridine orange as a transmembrane pH gradient probe.

    Science.gov (United States)

    Clerc, S; Barenholz, Y

    1998-05-15

    Monitoring the acidification of the internal space of membrane vesicles by proton pumps can be achieved easily with optical probes. Transmembrane pH gradients cause a blue-shift in the absorbance spectrum and the quenching of the fluorescence of the cationic dye acridine orange. It has been postulated that these changes are caused by accumulation and aggregation of the dye inside the vesicles. We tested this hypothesis using liposomes with transmembrane concentration gradients of ammonium sulfate as model system. Fluorescence intensity of acridine orange solutions incubated with liposomes was affected by magnitude of the gradient, volume trapped by vesicles, and temperature. These experimental data were compared to a theoretical model describing the accumulation of acridine orange monomers in the vesicles according to the inside-to-outside ratio of proton concentrations, and the intravesicular formation of sandwich-like piles of acridine orange cations. This theoretical model predicted quantitatively the relationship between the transmembrane pH gradients and spectral changes of acridine orange. Therefore, adequate characterization of aggregation of dye in the lumen of biological vesicles provides the theoretical basis for using acridine orange as an optical probe to quantify transmembrane pH gradients.

  11. Photocatalytic water splitting with acridine dyes: Guidelines from computational chemistry

    Science.gov (United States)

    Liu, Xiaojun; Karsili, Tolga N. V.; Sobolewski, Andrzej L.; Domcke, Wolfgang

    2016-01-01

    The photocatalytic splitting of water into Hrad and OHrad radicals in hydrogen-bonded chromophore-water complexes has been explored with computational methods for the chromophores acridine orange (AO) and benzacridine (BA). These dyes are strong absorbers within the range of the solar spectrum. It is shown that low-lying charge-transfer excited states exist in the hydrogen-bonded AOsbnd H2O and BAsbnd H2O complexes which drive the transfer of a proton from water to the chromophore, which results in AOHradsbnd OHrad or BAHradsbnd OHrad biradicals. The AOHrad and BAHrad radicals possess bright ππ∗ excited states with vertical excitation energies near 3.0 eV which are predissociated by a low-lying repulsive πσ∗ state. The conical intersections of the πσ∗ state with the ππ∗ excited states and the ground state provide a mechanism for the photodetachment of the H-atom by a second photon. Our results indicate that AO and BA are promising chromophores for water splitting with visible light.

  12. Synthesis and G-Quadruplex-Binding Properties of Defined Acridine Oligomers

    Directory of Open Access Journals (Sweden)

    Rubén Ferreira

    2010-01-01

    Full Text Available The synthesis of oligomers containing two or three acridine units linked through 2-aminoethylglycine using solid-phase methodology is described. Subsequent studies on cell viability showed that these compounds are not cytotoxic. Binding to several DNA structures was studied by competitive dialysis, which showed a clear affinity for DNA sequences that form G-quadruplexes and parallel triplexes. The fluorescence spectra of acridine oligomers were affected strongly upon binding to DNA. These spectral changes were used to calculate the binding constants (K. Log K were found to be in the order of 4–6.

  13. Synthesis, DNA Binding, and Antiproliferative Activity of Novel Acridine-Thiosemicarbazone Derivatives.

    Science.gov (United States)

    de Almeida, Sinara Mônica Vitalino; Lafayette, Elizabeth Almeida; da Silva, Lúcia Patrícia Bezerra Gomes; Amorim, Cézar Augusto da Cruz; de Oliveira, Tiago Bento; Ruiz, Ana Lucia Tasca Gois; de Carvalho, João Ernesto; de Moura, Ricardo Olímpio; Beltrão, Eduardo Isidoro Carneiro; de Lima, Maria do Carmo Alves; de Carvalho Júnior, Luiz Bezerra

    2015-01-01

    In this work, the acridine nucleus was used as a lead-compound for structural modification by adding different substituted thiosemicarbazide moieties. Eight new (Z)-2-(acridin-9-ylmethylene)-N-phenylhydrazinecarbothioamide derivatives (3a-h) were synthesized, their antiproliferative activities were evaluated, and DNA binding properties were performed with calf thymus DNA (ctDNA) by electronic absorption and fluorescence spectroscopies. Both hyperchromic and hypochromic effects, as well as red or blue shifts were demonstrated by addition of ctDNA to the derivatives. The calculated binding constants ranged from 1.74 × 10(4) to 1.0 × 10(6) M(-1) and quenching constants from -0.2 × 10(4) to 2.18 × 10(4) M(-1) indicating high affinity to ctDNA base pairs. The most efficient compound in binding to ctDNA in vitro was (Z)-2-(acridin-9-ylmethylene)-N- (4-chlorophenyl) hydrazinecarbothioamide (3f), while the most active compound in antiproliferative assay was (Z)-2-(acridin-9-ylmethylene)-N-phenylhydrazinecarbothioamide (3a). There was no correlation between DNA-binding and in vitro antiproliferative activity, but the results suggest that DNA binding can be involved in the biological activity mechanism. This study may guide the choice of the size and shape of the intercalating part of the ligand and the strategic selection of substituents that increase DNA-binding or antiproliferative properties. PMID:26068233

  14. Structural considerations on acridine/acridinium derivatives: Synthesis, crystal structure, Hirshfeld surface analysis and computational studies

    Science.gov (United States)

    Wera, Michał; Storoniak, Piotr; Serdiuk, Illia E.; Zadykowicz, Beata

    2016-02-01

    This article describes a detailed study of the molecular packing and intermolecular interactions in crystals of four derivatives of acridine, i.e. 9-methyl-, 9-ethyl, 9-bromomethyl- and 9-piperidineacridine (1, 2, 3 and 4, respectively) and three 10-methylacridinium salts containing the trifluoromethanesulphonate anion and 9-vinyl-, 9-bromomethyl, and 9-phenyl-10-methylacridinium cations (5, 6 and 7, respectively). The crystal structures of all of the compounds are stabilized by long-range electrostatic interactions, as well as by a network of short-range C-HṡṡṡO (in hydrates and salts 3 and 5-7, respectively), C-Hṡṡṡπ, π-π, C-Fṡṡṡπ and S-Oṡṡṡπ (in salts 5-7) interactions. Hirshfeld surface analysis shows that various intermolecular contacts play an important role in the crystal packing, graphically exhibiting the differences in spatial arrangements of the acridine/acridinium derivatives under scrutiny here. Additionally, computational methods have been used to compare the intermolecular interactions in the crystal structures of the investigated compounds. Computations have confirmed the great contribution of dispersive interactions for crystal lattice stability in the case of 9-substituted acridine and electrostatic interactions for the crystal lattice stability in the case of 9-substituted 10-methylacridinium trifluoromethanesulphonates. The value of crystal lattice energy and the electrostatic contribution in the crystal lattice energy of monohydrated acridine derivatives have confirmed that these compounds have behave as acridinium derivatives.

  15. Synthesis, DNA Binding, and Antiproliferative Activity of Novel Acridine-Thiosemicarbazone Derivatives

    Directory of Open Access Journals (Sweden)

    Sinara Mônica Vitalino de Almeida

    2015-06-01

    Full Text Available In this work, the acridine nucleus was used as a lead-compound for structural modification by adding different substituted thiosemicarbazide moieties. Eight new (Z-2-(acridin-9-ylmethylene-N-phenylhydrazinecarbothioamide derivatives (3a–h were synthesized, their antiproliferative activities were evaluated, and DNA binding properties were performed with calf thymus DNA (ctDNA by electronic absorption and fluorescence spectroscopies. Both hyperchromic and hypochromic effects, as well as red or blue shifts were demonstrated by addition of ctDNA to the derivatives. The calculated binding constants ranged from 1.74 × 104 to 1.0 × 106 M−1 and quenching constants from −0.2 × 104 to 2.18 × 104 M−1 indicating high affinity to ctDNA base pairs. The most efficient compound in binding to ctDNA in vitro was (Z-2-(acridin-9-ylmethylene-N- (4-chlorophenyl hydrazinecarbothioamide (3f, while the most active compound in antiproliferative assay was (Z-2-(acridin-9-ylmethylene-N-phenylhydrazinecarbothioamide (3a. There was no correlation between DNA-binding and in vitro antiproliferative activity, but the results suggest that DNA binding can be involved in the biological activity mechanism. This study may guide the choice of the size and shape of the intercalating part of the ligand and the strategic selection of substituents that increase DNA-binding or antiproliferative properties.

  16. Preparation of acridine orange-doped silica nanoparticles for pH measurement

    Energy Technology Data Exchange (ETDEWEB)

    Liu, Jinshui, E-mail: jsliu@sina.com; Zang, Lingjie; Wang, Yiru; Liu, Guoning

    2014-03-15

    Acridine orange was first encapsulated into silica shell via a facile reverse microemusion method to built core–shell fluorescent nanoparticles. The nanoparticles are all in spherical shape and have a narrow size distribution, and its application as a optical pH sensor has been demonstrated. This novel sensor is based on the pH-dependent fluorescence intensities of acridine orange in different pH value. The fluorescence intensity of acridine orange-doped silica nanoparticles was decreased by increasing pH value. Under optimum conditions, the changes of fluorescence intensity were proportional to the pH value in the range of 8.00–10.90. In addition, the sensor can be easily separated by centrifugation and adds no pollution to the environment compared to the free dyes. Furthermore, the effects of ionic strength and co-existing substances were proved to have little influence on the determination of pH. The sensor has been successfully applied to determine the pH of two artificial samples. Hence, the core–shell fluorescent nanoparticles show potential for practical application. -- Highlights: • Acridine orange was encapsulated into silica shell via a facile reverse microemusion method to built core–shell fluorescent nanoparticles. • The fluorescence intensity of acridine orange-doped silica nanoparticles was decreased by increasing pH value. • Its can be used as an optical pH sensor. • The sensor can be easily separated by centrifugation and adds no pollution to the environment compared to the free dyes. • The sensor has been successfully applied to determine the pH of artificial samples.

  17. Synthesis, cytotoxicity and structure-activity relationships between ester and amide functionalities in novel acridine-based platinum(II) complexes.

    Science.gov (United States)

    Bouyer, Florence; Moretto, Johnny; Pertuit, David; Szollosi, Anna; Lacaille-Dubois, Marie-Aleth; Blache, Yves; Chauffert, Bruno; Desbois, Nicolas

    2012-05-01

    In order to improve the pharmacological profile of the anticancer drug cisplatin, several new acridine-based tethered (ethane-1,2-diamine)platinum(II) complexes connected by a polymethylene chain were synthetized. Activity-structure relationship between amide or ester functionalities was explored by changing acridine-9-carboxamide into acridine-9-carboxylate chromophore. The in vitro cytotoxicity of these new complexes was assessed in human colic HCT 116, SW480 and HT-29 cancer cell lines. Series of complexes bearing the acridine-9-carboxylate chromophore displayed higher cytotoxic effect than acridine-9-carboxamide complexes, with gradual effect according to the size of the polymethylene linker. PMID:22459174

  18. Morphology Dependent Photocatalytic Activity of α-MoO3 Nanostructures Towards Mutagenic Acridine Orange Dye.

    Science.gov (United States)

    2015-06-01

    The morphological evolutions of orthorhombic molybdenum oxide nanostructures with high crystalline nature have been successfully synthesized by combining low-temperature sol-gel and annealing processes. Strong influence of gelation temperature is a factor facilitated to control the material morphology. Morphological transformations like nanospheres, nanoplatelets, mixtures of hexagonal platelets, and one-dimensional nanobars were obtained. The possible morphological formation mechanism has been proposed as a self-assemble process of nucleation and a mechanism for particle growth by Ostwald ripening. The as-prepared nanostructures were recognized as photocatalysts for the degradation of Acridine Orange under Ultra Violet light. The obtained mixed morphology (hexagonal nanoplatelets and nanobars) showed a high photocatalytic property to degrade mutagenic Acridine Orange dye. Moreover, they could be easily recycled without changing the photocatalytic activity due to their 1-Dimensional and 2-Dimensional nanostructure property. PMID:26369043

  19. Microscopic observation of progressive immobilization of leishmania promastigotes in acridine orange stain.

    OpenAIRE

    G.S. Barreca; Berlinghieri, M C; F. Foti; G. Matera; Foca, A

    1997-01-01

    To rapidly isolate Leishmania donovani promastigotes in samples from Novy-MacNeal-Nicolle (NNN) cultures, a method of staining with acridine orange was developed. Such vital staining combines the advantages of direct microscopic examination (e.g., observation of motility) with more accurate cytological and structural imaging of the stained parasites (usually obtained by Giemsa staining). Progressive immobilization of Leishmania promastigotes associated with a change in fluorescence color was ...

  20. Solvent-free preparation of co-crystals of phenazine and acridine with vanillin

    International Nuclear Information System (INIS)

    Co-crystals of phenazine and acridine with vanillin have been obtained by solvent-free reaction or thermal treatment of the solid reactants: their structures, thermal behaviour and eutectic formation have been investigated via single crystal X-ray diffraction, differential scanning calorimetry (DSC), variable temperature X-ray powder diffraction and hot-stage microscopy (HSM). Polymorph screening of the reagents has also been carried out.

  1. Systematic colocalization errors between acridine orange and EGFP in astrocyte vesicular organelles

    OpenAIRE

    Nadrigny, F.; Li, D.; Kemnitz, K; Ropert, N.; Koulakoff, A; Rudolph, S.; Vitali, M.; Giaume, C.; Kirchhoff, F.; Oheim, M

    2007-01-01

    Dual-color imaging of acridine orange (AO) and EGFP fused to a vesicular glutamate transporter or the vesicle-associated membrane proteins 2 or 3 has been used to visualize a supposedly well-defined subpopulation of glutamatergic astrocytic secretory vesicles undergoing regulated exocytosis. However, AO metachromasy results in the concomitant emission of green and red fluorescence from AO-stained tissue. Therefore, the question arises whether AO and EGFP fluorescence can be distinguished reli...

  2. Solvent-free preparation of co-crystals of phenazine and acridine with vanillin

    Energy Technology Data Exchange (ETDEWEB)

    Braga, Dario, E-mail: dario.braga@unibo.it [Dipartimento di Chimica ' G.Ciamician' , Universita degli studi di Bologna, Via Selmi 2, 40126 Bologna (Italy); Grepioni, Fabrizia; Maini, Lucia; Mazzeo, Paolo P.; Rubini, Katia [Dipartimento di Chimica ' G.Ciamician' , Universita degli studi di Bologna, Via Selmi 2, 40126 Bologna (Italy)

    2010-08-10

    Co-crystals of phenazine and acridine with vanillin have been obtained by solvent-free reaction or thermal treatment of the solid reactants: their structures, thermal behaviour and eutectic formation have been investigated via single crystal X-ray diffraction, differential scanning calorimetry (DSC), variable temperature X-ray powder diffraction and hot-stage microscopy (HSM). Polymorph screening of the reagents has also been carried out.

  3. Nonlinear phenomena of acridine orange in inorganic glasses at nanosecond scale

    Science.gov (United States)

    Gaponenko, S. V.; Gribkovskii, V. P.; Zimin, L. G.; Lebed, V. Yu.; Malinovskii, I. E.; Graham, S.; Klingshirn, C.; Eyal, M.; Brusilovsky, D.; Reisfeld, R.

    1993-04-01

    Nonlinear optical behavior of acridine orange dye has been studied in lead-tin-flouride glass. We found that this material possess nonlinear saturable absorption and power-dependent lifetimes, both on nanosecond time scale. This short response is explained by an efficient S-T transfer induced by the heavy atoms of the glass. The glass has a good potential as a nonlinear material on a nanosecond time scale.

  4. DNA binding, anti-tumour activity and reactivity toward cell thiols of acridin-9-ylalkenoic derivatives

    Indian Academy of Sciences (India)

    O Salem; M Vilkova; J Plsikova; A Grolmusova; M Burikova; M Prokaiova; H Paulikova; J Imrich; M Kozurkova

    2015-05-01

    In this paper, we describe the synthesis, biochemical properties and biological activity of a series of new 9-substituted acridine derivatives with a reactive alkene moiety: 9-[(E)-2-phenylethenyl] acridine (1) and methyl (2E)-3-(acridin-9-yl)-prop-2-enoate (2). The interaction of derivatives 1 and 2 with calf thymus DNA was investigated using UV-Vis, fluorescence and circular dichroism spectroscopy. The binding constants K were estimated as being in the range of 1.9 to 7.1 × 105 M−1, and the percentage of hypochromism was found to be 40–57% (from spectral titration). UV-Vis, fluorescence, and CD measurements indicate that the compounds were effective DNA-intercalating agents. Electrophoretic separation proved that ligands 1 and 2 relaxed topoisomerase I at a concentration of 5 M. Ester 2 was shown to have a stronger cytostatic effect on leukemia cell line L1210 than alkene 1. The incubation of ligands 1 and 2 with the ovarian carcinoma cell line A2780 confirmed their extensive cytotoxic effects, an effect which was particularly pronounced in the case of ligand 2. Cytotoxicity tests against A2780 cells demonstrate that a conjugate of compound 2 with -cysteine (3) is less cytotoxic than compound 2, especially at concentrations greater than 10 M.

  5. Inhibition of RNA synthesis in vitro by acridines--relation between structure and activity.

    Science.gov (United States)

    Piestrzeniewicz, M K; Wilmańska, D; Studzian, K; Szemraj, J; Czyz, M; Denny, W A; Gniazdowski, M

    1998-01-01

    The effects of acridine derivatives (proflavine and 2,7-dialkyl derivatives, diacridines and triacridines, 9-aminoacridine carboxamides, and 9-anilinoacridine, amsacrine and its congeners) on overall RNA synthesis in vitro, on synthesis of initiating oligonucleotides and the binding of the enzyme to DNA were studied. The primary mechanism of action is related to inhibition of the enzyme binding to DNA. The acridines (intercalating or non-intercalating and bis-intercalating ligands) assayed here differ in the properties of their complexes with DNA. Correlation is generally observed between inhibition of RNA synthesis in vitro and cytotoxicity in cell cultures for di- and triacridines and 9-aminoacridine carboxamide derivatives. No relationship was found between the effect on RNA polymerase system and biological effects for amsacrine and its derivatives in contrast to the other series of acridines studied here. The aniline ring seems to decrease the inhibitory potency of a ligand. The discrepancy between the biological effect and RNA synthesis inhibition may be due to a different mechanism of cytotoxicity action of amsacrine which is a potent topoisomerase II poison. PMID:9679327

  6. Basicities of some 9-substituted acridine-4-carboxamides: A density functional theory (DFT) calculation

    Indian Academy of Sciences (India)

    Raghab Parajuli; C Medhi

    2004-06-01

    Acid-base properties of drugs are important in understanding the behaviour of these compounds under physiological condition. In order to understand such behaviour the proton affinities of acridine 4-carboxamides with substitution (R) at the 9-position are theoretically studied, and considered for the basic sites of both the heterocyclic ring as well as side chain nitrogens. In 9-amino acridine 4-carboxamide, the -NH2 group is observed to be an additional basic site. The heterocyclic nitrogen of substituted carboxamides (R = -NH2, -O-methyl, -O-ethyl, and -O-phenyl) is more basic than the side chain nitrogen, however, side chain nitrogen corresponds to more basic site for some carboxamides (R = -OH and -Cl) and the -NH2 group represents the least basic site of 9-amino acridine 4-carboxamide. In addition to presenting the basicities of these drugs an indication of another hydrogen-bond between heterocyclic ring N and carboxamide chain O is observed. The difference of basicities with substituents at 9-position are very narrow and carboxamides with substituents at 9-position are found to be suitable for studying intramolecular H-bonds between the heterocyclic N and carboxamide O. The resultant stabilization of a configuration due to such H-bonding is determined.

  7. Supra-molecular inter-actions in a 1:1 co-crystal of acridine and 3-chloro-thio-phene-2-carb-oxy-lic acid.

    Science.gov (United States)

    Prajina, Olakkandiyil; Thomas Muthiah, Packianathan; Perdih, Franc

    2016-05-01

    In the title co-crystal, C5H3ClO2S·C13H9N, the components inter-act with each other via an O-H⋯N hydrogen bond. Acridine-acridine stacking, thio-phene-thio-phene stacking and acridine-thio-phene C-H⋯π inter-actions also occur in the crystal. PMID:27308013

  8. Thiol-dependent inhibition of RNA synthesis in vitro by acridines: structure-inhibition relationships.

    Science.gov (United States)

    Gniazdowski, M; Szmigiero, L; Wilmańska, D

    1982-01-01

    In the presence of sulfhydryl compounds an anticancer drug, 1-nitro-9-aminoalkylacridine derivative, forms with DNA irreversible, probably covalent, complexes of decreased template properties. Five 9-substituted 1-nitro-9-aminoacridine derivatives of cytostatic activity show irreversible thiol-dependent inhibitory effects on the RNA synthesis in vitro system while equal inhibition is observed both in the presence and in the absence of dithiothreitol with biologically inactive analogues of nitrocrine. In the absence of sulfhydryl compounds the inhibition depends on the planarity of the acridine ring. Hence, both 1-nitro-9-aminoalkylacridine and tetrahydroacridine derivatives show low inhibitory effect. PMID:6174208

  9. Acridin-9-ylmethoxycarbonyl (Amoc): A New Photochemically Removable Protecting Group for Alcohols

    Institute of Scientific and Technical Information of China (English)

    WANG Hong-Bo; TANG Wen-Jian; YU Jing-Yu; SONG Qin-Hua

    2006-01-01

    Synthesis and photochemistry of acridin-9-ylmethoxycarbonyl (Amoc) as a new photochemically removable protecting group for alcohols were described. Three carbonates of alcohols 1-3 were synthesized through condensation of 9-hydroxymethylacridine and chloroformates of alcohols, including benzyl alcohol, phenethyl alcohol and one galactose derivative. The photolysis of protected alcohols can efficiently release the corresponding alcohol in the efficiencies (Qu1ε) of 100-200 (quantum yield Qu1=0.011-0.023, and molar absorptivity ε=9.1 × 103-9.8 × 103 mol-1·L·cm-1) under 360 nm light.

  10. Kinetics and Adsorption Isotherms Studies of Acridine Orange Dye from Aqueous Solution by Activated Charcoal

    OpenAIRE

    2N. Qamar; R. Azmat; Naz, R.; Malik, B.

    2014-01-01

    The goal of this research is to evaluate the efficiency of charcoal as low coast and effective adsorbent for acridine orange (a cationic dye) from aqueous solution at room temperature. Effect of initial pH (2-8), shaking time (5min. - 1hour), adsorbent dose (0.1gm- 0.9gm) and dye concentration (37mg/30ml-185mg/30ml) were investigated. Results demonstrated that charcoal act as good adsorbent for the removal AO where 99.15% of the dye was adsorbed within 30 minutes. For the maximum dye removal ...

  11. Fluorescence enhancement of acridine orange in a water solution by Au nanoparticles

    Institute of Scientific and Technical Information of China (English)

    2010-01-01

    The surface enhanced fluorescence effect of acridine orange fluorophore in the proximity of Au nanoparticles has been investigated experimentally in the system of aqueous solution.Significant enhancement of the fluorescence intensity was observed when the system was excited with 532 nm or 442 nm CW lasers.The influence of the distances between neighboring Au particles as well as that between the fluorophore molecules and the Au surface were explored experimentally.The results demonstrated that a compact distribution of metallic particles was able to produce stronger fluorescence enhancement.Proper separation between the fluorophore molecules and the metal surface was favorable for a better enhancement.

  12. Highly selective and sensitive fluorescence detection of Zn(2+) and Cd(2+) ions by using an acridine sensor.

    Science.gov (United States)

    Visscher, A; Bachmann, S; Schnegelsberg, C; Teuteberg, T; Mata, R A; Stalke, D

    2016-04-01

    Fluorescence spectroscopy investigations of the new acridine derivative bis(N,N-dimethylaminemethylene)acridine (3) show remarkable selectivity and sensitivity towards Zn(2+) and Cd(2+) ions in methanol and for the latter even in water. Through the chelation of the metal ions the present PET effect is quenched, significantly enhancing the emission intensity of the fluorophore. In solution, the bonding situation is studied by fluorescence and NMR spectroscopy, as well as ESI-TOF mass-spectrometry measurements. The solid state environment is investigated by X-ray diffraction and computational calculations. Here, we can show the complexation of the zinc and cadmium ions by the methylene bridged amine receptors as well as by the nitrogen atom of the acridine system. PMID:26928871

  13. Cytotoxic and DNA-damaging properties of N-[2-(dimethylamino)ethyl]acridine-4-carboxamide (DACA) and its analogues.

    Science.gov (United States)

    Pastwa, E; Ciesielska, E; Piestrzeniewicz, M K; Denny, W A; Gniazdowski, M; Szmigiero, L

    1998-08-01

    An antitumor drug N-[2-(dimethylamino)ethyl]acridine-4-carboxamide (DACA) and its three close structural analogs N-[2-(hydroxyethylamino)ethyl]acridine-4-carboxamide (DACAH), N-[2-(dimethylamino)ethyl]-9-aminoacridine-4-carboxamide (amino-DACA), and N-[2-(hydroxyethylamino)ethyl]-9-aminoacridine-4-carboxamide (amino-DACAH) were studied for their ability to inhibit RNA synthesis in vitro and to form topoisomerase II-mediated DNA lesions in relation to cell-killing activity. All tested compounds induced chromatin lesions characteristic of topoisomerase II-blocking drugs (DNA breaks and DNA-protein cross-links) in treated cells, but were much less active than reference antileukemic acridine m-AMSA (4'-(9-acridinylamino)-methanesulfon-m-anisidide). The ability to form these lesions was dependent on the structure of the 4-carboxamide side-chain, which seems to be an important factor affecting the drug transport rate through cell membrane. A 4-carboxamide chain with an N-2-(dimethylamino)ethyl moiety resulted in more efficient transport through cell membranes, higher cytotoxicity, and DNA-damaging activity. The mode of action of acridine-4-carboxamides was further elucidated by their incubation with cells in the presence of antitopoisomerase II agents of a known mechanism of inhibition. These were: bisdioxopiperazine (ICRF-187), a catalytic inhibitor of topoisomerase II, and etoposide (VP-16), an inducer of a cleavable complex of the enzyme with DNA. The cytotoxicity of DACA and its analogs was not antagonized by preincubating cells with ICRF-187. All tested acridines protected cells against DNA breakage induced by VP-16, but the extent of protection varied significantly. Amino-DACA, which easily penetrates cell membrane, fully inhibited DNA break formation, whereas other analogs exhibited a low degree of protection when used at high concentration. Our results suggest that the acridine-4-carboxamides discussed here are poor topoisomerase II poisons and that this enzyme

  14. Interaction and sonodynamic damage activity of acridine red (AD-R) to bovine serum albumin (BSA)

    Energy Technology Data Exchange (ETDEWEB)

    Chen, Dandan; Xie, Jinhui; Wu, Qiong; Fan, Ping; Wang, Jun, E-mail: wangjun888tg@126.com

    2015-04-15

    The sonodynamic therapy (SDT) has become an attractive antitumor treatment method in recent years, but the selection of sonosensitizer, mechanism of damage biomolecule and kind of reactive oxygen species (ROS) generated during sonodynamic process have not been investigated in detail. In this paper, the acridine red (AD-R), as a sonosensitizer, combining with ultrasonic irradiation to damage bovine serum albumin (BSA) was investigated. At first, the interaction of AD-R to BSA molecules in aqueous solution was studied by fluorescence spectroscopy. As judged from the experimental results, the quenching mechanism of BSA fluorescence belongs to a static process. Synchronous fluorescence spectra demonstrate that the binding and damage sites to BSA molecules are mainly on the tryptophan residues. The generation and kind of generated ROS were also estimated by the method of oxidation and extraction photometry. This paper may offer some valuable references for the study of the sonodynamic activity and application of AD-R in SDT for tumor treatment. - Highlights: ●Acridine red (AD-R) is used to study interaction with BSA. ●Spectroscopy is used to study sonodynamic damage activity of AD-R to BSA. ●Generation of ROS caused by AD-R under ultrasonic irradiation was determined.

  15. Evaluation of chromatin condensation in human spermatozoa: a flow cytometric assay using acridine orange staining.

    Science.gov (United States)

    Golan, R; Shochat, L; Weissenberg, R; Soffer, Y; Marcus, Z; Oschry, Y; Lewin, L M

    1997-01-01

    The quality of sperm chromatin is an important factor in fertilization and is especially critical where one spermatozoon is artificially selected for fertilizing an egg (as in intracytoplasmic sperm injection). In this study, flow cytometry after staining of human spermatozoa with Acridine Orange was used to study chromatin structure. A method is described for estimating the percentage of cells in a human sperm sample that have completed epididymal maturation in regard to chromatin condensation. Of the 121 samples of the semen that were examined, nine contained a higher percentage of hypocondensed spermatozoa and six samples contained elevated amounts of hypercondensed spermatozoa. In addition to aberrancies in chromatin condensation other defects showed up as satellite populations of spermatozoa with higher than normal ratios of red/green fluorescence after Acridine Orange staining. Such defects were found in 15 semen samples. The use of swim-up and Percoll gradient centrifugation methods was shown to improve the percentage of spermatozoa with normal chromatin structure in some samples with poor initial quality.

  16. Pyrrolidine-Acridine hybrid in Artemisinin-based combination: a pharmacodynamic study.

    Science.gov (United States)

    Pandey, Swaroop Kumar; Biswas, Subhasish; Gunjan, Sarika; Chauhan, Bhavana Singh; Singh, Sunil Kumar; Srivastava, Kumkum; Singh, Sarika; Batra, Sanjay; Tripathi, Renu

    2016-09-01

    Aiming to develop new artemisinin-based combination therapy (ACT) for malaria, antimalarial effect of a new series of pyrrolidine-acridine hybrid in combination with artemisinin derivatives was investigated. Synthesis, antimalarial and cytotoxic evaluation of a series of hybrid of 2-(3-(substitutedbenzyl)pyrrolidin-1-yl)alkanamines and acridine were performed and mode of action of the lead compound was investigated. In vivo pharmacodynamic properties (parasite clearance time, parasite reduction ratio, dose and regimen determination) against multidrug resistant (MDR) rodent malaria parasite and toxicological parameters (median lethal dose, liver function test, kidney function test) were also investigated. 6-Chloro-N-(4-(3-(3,4-dimethoxybenzyl)pyrrolidin-1-yl)butyl)-2-methoxyacridin-9-amine (15c) has shown a dose dependent haem bio-mineralization inhibition and was found to be the most effective and safe compound against MDR malaria parasite in Swiss mice model. It displayed best antimalarial potential with artemether (AM) in vitro as well as in vivo. The combination also showed favourable pharmacodynamic properties and therapeutic response in mice with established MDR malaria infection and all mice were cured at the determined doses. The combination did not show toxicity at the doses administered to the Swiss mice. Taken together, our findings suggest that compound 15c is a potential partner with AM for the ACT and could be explored for further development. PMID:27230403

  17. Determination of sex by exfoliative cytology using acridine orange confocal microscopy: A short study

    Directory of Open Access Journals (Sweden)

    D Shyam Prasad Reddy

    2012-01-01

    Full Text Available Context: Establishing individuality is an imperative aspect in any investigation procedure. Sometimes, in identifying an individual, it becomes necessary to determine the sex of that particular individual. Combining rapidity with reliability, an innovative idea has been put forward using a confocal microscope in exfoliative cytology. In the present study, we have determined the sex of the individual from buccal mucosal scrapings. The exfoliative cells were observed for Barr bodies under a confocal microscope, and the percentage of Barr-body-positive cells was determined. Aims: The main objective of this study is to assess confocal microscopy for the determination of sex by observing Barr bodies in the exfoliative cells of both men and women. Settings and Design: Samples of buccal mucosa smears were made followed by acridine orange staining. The stained slides were observed under a confocal microscope and the data obtained was subjected for statistical analysis, especially for mean and standard deviation. Materials and Methods: Samples of buccal mucosa smears from 20 men and 20 women were obtained by scraping with flat wooden sticks (exfoliative cytology. The smears were fixed in 100% alcohol for 15 min, followed by acridine orange (AO staining as described by Von Bertalanffy et al. Smears stained with AO were examined under a confocal microscope and the percentage of Barr-body-positive cells was determined. Statistical Analysis Used: Data obtained was subjected for statistical analysis, especially for mean and standard deviation. Results: Two non-overlapping ranges for the percentage of Barr-body-positive cells have been obtained for men and women. It was observed that in the male samples, the percentage of Barr-body-positive cells ranged from 0-3%. In the female samples, the percentage of Barr-body-positive cells ranged from 18-72%, and all the females showed the presence of Barr bodies. Conclusion: The study showed that the presence of Barr

  18. Photoabsorption of Acridine Yellow and Proflavin Bound to Human Serum Albumin Studied by Means of Quantum Mechanics/Molecular Dynamics

    DEFF Research Database (Denmark)

    Aidas, Kestutis; Olsen, Jógvan Magnus Haugaard; Kongsted, Jacob;

    2013-01-01

    Attempting to unravel mechanisms in optical probing of proteins, we have performed pilot calculations of two cationic chromophores—acridine yellow and proflavin—located at different binding sites within human serum albumin, including the two primary drug binding sites as well as a heme binding site...

  19. Comparison of acridine orange and Gram stains for detection of microorganisms in cerebrospinal fluid and other clinical specimens.

    OpenAIRE

    Lauer, B. A.; Reller, L B; Mirrett, S

    1981-01-01

    Acridine orange, a fluorochrome strain, is potentially superior to the Gram stain in the direct microscopic examination of clinical specimens because it gives striking differential staining between bacteria and background cells and debris. Its value in clinical laboratories was evaluated by testing 209 cerebrospinal fluids and 288 other body fluids, tissues, and exudates by both techniques. Smears were made in duplicate, fixed with methanol, stained, and examined without knowledge of the resu...

  20. Evaluation of new iodinated acridine derivatives for targeted radionuclide therapy of melanoma using 125I, an Auger electron emitter.

    Science.gov (United States)

    Gardette, Maryline; Papon, Janine; Bonnet, Mathilde; Desbois, Nicolas; Labarre, Pierre; Wu, Ting-Dee; Miot-Noirault, Elisabeth; Madelmont, Jean-Claude; Guerquin-Kern, Jean-Luc; Chezal, Jean-Michel; Moins, Nicole

    2011-12-01

    The increasing incidence of melanoma and the lack of effective therapy on the disseminated form have led to an urgent need for new specific therapies. Several iodobenzamides or analogs are known to possess specific affinity for melanoma tissue. New heteroaromatic derivatives have been designed with a cytotoxic moiety and termed DNA intercalating agents. These compounds could be applied in targeted radionuclide therapy using (125)I, which emits Auger electrons and gives high-energy, localized irradiation. Two iodinated acridine derivatives have been reported to present an in vivo kinetic profile conducive to application in targeted radionuclide therapy. The aim of the present study was to perform a preclinical evaluation of these compounds. The DNA intercalating property was confirmed for both compounds. After radiolabeling with (125)I, the two compounds induced in vitro a significant radiotoxicity to B16F0 melanoma cells. Nevertheless, the acridine compound appeared more radiotoxic than the acridone compound. While cellular uptake was similar for both compounds, SIMS analysis and in vitro protocol showed a stronger affinity for melanin with acridone derivative, which was able to induce a predominant scavenging process in the melanosome and restrict access to the nucleus. In conclusion, the acridine derivative with a higher nuclear localization appeared a better candidate for application in targeted radionuclide therapy using (125)I. PMID:20567996

  1. Unexpected regiospecific formation and DNA binding of new 3-(acridin-9-yl)methyl-2-iminothiazolidin-4-ones

    Indian Academy of Sciences (India)

    Ján Imrich; Danica Sabolová; Mária Vilková; Júlia Kudláčová

    2016-02-01

    New 3-(acridin-9-yl)methyl-2-substituted imino-1,3-thiazolidin-4-ones were regiospecifically synthesized from unstable (acridin-9-yl)methyl thioureas and methyl bromoacetate (MBA) or bromoacetyl bromide (BAB). Unexpected formation of only one thiazolidinone regioisomer with both the reagents was due to a new mechanism involving a transient spiro 9,10-dihydroacridine intermediate. These results are in contrast with the reactions of acridin-9-yl thioureas with MBA/BAB that afforded two different thiazolidinone regioisomers with these reagents. UV-vis titrations, CD spectra, and fluorescence quenching have shown that new products intercalated into calf thymus (CT) DNA, and displaced ethidium bromide (EB) from a CT DNA–EB complex. Intrinsic binding constants, , and Stern-Volmer constants, , were found in the range 0.79✕105 – 2.85✕105 M−1 and 17950 – 3360M−1, respectively. The strongest binding affinity was found for an electrondonated 2-(4-methoxyphenylimino) thiazolidinone. Additional evidence for DNA intercalation was obtained from thermal denaturation studies. Gel electrophoresis has proven that thiazolidinone products nicked the supercoiled plasmid DNA in 5.0 M concentration.

  2. Enhanced fluorescence quenching in an acridine orange - alizarin red system through matrine and its analytical application

    Science.gov (United States)

    Wei, Xiaoling; Wang, Xiaojun; Gong, Qi; Wang, Lisheng; Zhou, Shiwu

    2015-01-01

    This study shows that alizarin red (AR) only slightly quenched fluorescence for acridine orange (AO) in an AR/AO mixed solution at pH = 5-6. The reduced fluorescent signal was closely and linearly associated with the level of MT added to the system, which is the basis for a new quantitative MT assay method using the fluorescence quenching reaction in the AO-AR system. The results show that under optimal conditions, this method had a 14.9-43.5 mg L-1 linear detection range with a 1.38 mg L-1 detection limit and 1.24% precision. In addition, this method was used to determine the MT levels in the commercially available MT-containing pesticides and suppositories, which showed a 96.6-103% recovery. Therefore, this method has multiple advantages, including simple and fast operation, high accuracy and low cost. Moreover, herein, we investigated the underlying mechanism in-depth using an ultraviolet (UV) spectroscopic technique.

  3. Structural relaxation of acridine orange dimer in bulk water and inside a single live lung cell

    Science.gov (United States)

    Chowdhury, Rajdeep; Nandi, Somen; Halder, Ritaban; Jana, Biman; Bhattacharyya, Kankan

    2016-02-01

    Structural relaxation of the acridine orange (AO) dimer in bulk water and inside a single live lung cell is studied using time resolved confocal microscopy and molecular dynamics (MD) simulations. The emission maxima ( λem max ˜ 630 nm) of AO in a lung cancer cell (A549) and a non-cancer lung fibroblast cell (WI38) suggest that AO exists as a dimer inside the cell. Time-dependent red shift in emission maximum indicates dynamic relaxation of the AO dimer (in the excited state) with a time constant of 500-600 ps, both in bulk water and inside the cell. We have calculated the equilibrium relaxation dynamics of the AO dimer in the ground state using MD simulations and found a slow component of time scale ˜350 ps. The intra- and inter-molecular components of the total relaxation dynamics of the AO dimer reveal the presence of a slow component of the order of a few hundred picoseconds. Upon restricting intra-molecular dye dynamics by harmonic constraint between AO monomers, the slow component vanishes. Combining the experimental observations and MD simulation results, we ascribe the slow component of the dynamic relaxation of the AO dimer to the structural relaxation, namely, fluctuations in the distance between the two monomers and associated fluctuation in the number of water molecules.

  4. Acridine orange staining reaction as an index of physiological activity in Escherichia coli

    Science.gov (United States)

    McFeters, G. A.; Singh, A.; Byun, S.; Callis, P. R.; Williams, S.

    1991-01-01

    The assumption that the acridine orange (AO) color reaction may be used as an index of physiological activity was investigated in laboratory grown Escherichia coli. Spectrofluorometric observations of purified nucleic acids, ribosomes and the microscopic color of bacteriophage-infected cells stained with AO confirmed the theory that single-stranded nucleic acids emit orange to red fluorescence while those that are double-stranded fluoresce green in vivo. Bacteria growing actively in a rich medium could be distinguished from cells in stationary phase by the AO reaction. Cells from log phase appeared red, whereas those in stationary phase were green. However, this differentiation was not seen when the bacteria were grown in a minimal medium or when a variation of the staining method was used. Also, shifting bacteria in stationary phase to starvation conditions rapidly changed their AO staining reaction. Boiling and exposure to lethal concentrations of azide and formalin resulted in stationary-phase cells that appeared red after staining but bacteria killed with chlorine remained green. These findings indicate that the AO staining reaction may be suggestive of physiological activity under defined conditions. However, variables in staining and fixation procedures as well as uncertainties associated with mixed bacterial populations in environmental samples may produce results that are not consistent with the classical interpretation of this reaction. The importance of validating the putative physiological implications of this staining reaction is stressed.

  5. Kinetics and Adsorption Isotherms Studies of Acridine Orange Dye from Aqueous Solution by Activated Charcoal

    Directory of Open Access Journals (Sweden)

    *N. Qamar

    2014-12-01

    Full Text Available The goal of this research is to evaluate the efficiency of charcoal as low coast and effective adsorbent for acridine orange (a cationic dye from aqueous solution at room temperature. Effect of initial pH (2-8, shaking time (5min. - 1hour, adsorbent dose (0.1gm- 0.9gm and dye concentration (37mg/30ml-185mg/30ml were investigated. Results demonstrated that charcoal act as good adsorbent for the removal AO where 99.15% of the dye was adsorbed within 30 minutes. For the maximum dye removal efficiency (100%, optimum conditions were obtained at pH 8 (99.24%, adsorbent dose of 0.9g and dye concentration of 185 mg with charcoal. Kinetics of adsorption was investigated as well as Langmuir and Freundlich isotherms were employed to describe equilibrium studies. The Langmuir adsorption isotherms models and pseudo second order kinetics fitted the experimental data best with high regression coefficient R2. The results of the present studies points to the potential of charcoal as an effective adsorbent for the removal of dye from contaminated water sources.

  6. Flow cytometry reticulocyte counting using acridine orange: validation of a new protocol

    Directory of Open Access Journals (Sweden)

    Karina Augusta Viana

    2014-06-01

    Full Text Available Introduction: Currently, the reticulocyte counting is a challenge for clinical laboratories in Brazil, mainly for the ordinary ones, which still use the manual method. This method has some limitations, since it consists of a laborious method, time consuming, with low accuracy. Objectives: This study has developed and evaluated the performance of a New Laboratory Protocol for flow cytometry (FC reticulocytes counting using acridine orange (AO as dye, aiming to standardize a more precise, easy, fast implementation, and low cost protocol. After standardization of the New Protocol (FC/AO, it was compared with the manual method. The results were analyzed according to the recommendations of the National Committee for Clinical Laboratory Standards (NCCLS, now known as Clinical and Laboratory Standards Institute (CLSI, to evaluate the interchangeability of methods in linear regression analysis and paired t test, besides other quality control tests. Conclusion: Based on these results concerning to the correlation between the methods and the tests related to quality control, we can admit that FC/AO for reticulocyte counting shows undeniable advantages when compared to the preexisting manual method.

  7. Linear sweep polarographic determination of nucleic acids using acridine orange as a bioprobe

    Directory of Open Access Journals (Sweden)

    WEI SUN

    2007-11-01

    Full Text Available The interaction of acridine orange (AO with double-stranded (ds DNA in aqueous solution was investigated by linear sweep polarography (LSP on a dropping mercury working electrode (DME. In pH 2.5 Britton–Robinson (B–R buffer solution, AO had a sensitive linear sweep polarographic reductive peak at –0.89 V (vs. SCE, which could be greatly inhibited by the addition of dsDNA, with a positive shift of the peak potential. Based on the decrease of the reductive peak current, a new quantitative electrochemical determination method for dsDNA was developed with a linear range of 2.0−20.0 mg l-1 and the linear regression equation: ΔIp” (nA = 111.90 C (mg l-1+125.32 (n = 9, γ = 0.997. The influences of commonly co-existing substances, such as metal ions, amino acid, etc., on the determination were also investigated. The method is sensitive, rapid and simple with good selectivity. The new proposed method was further applied to the detection of RNA and three synthetic samples containing dsDNA with satisfactory results. The binding number and the equilibrium constant between dsDNA and AO were calculated by an electrochemical method.

  8. Comparative study of subculture, Gram staining and acridine orange staining for early detection of positive blood cultures.

    OpenAIRE

    Mascart, G; Bertrand, F.; Mascart, P.

    1983-01-01

    In view of the importance of a rapid aetiological diagnosis in septicaemia, we compared the results of subculture, Gram staining and acridine orange staining in the detection of positive blood cultures. The study was based on 1013 blood cultures of which 138 were positive by culture. The three techniques were applied 12 h after the specimen was taken in 210 instances, at 24 h in 540 instances and after 48 h in 525. We were able to demonstrate the value of direct examination. Staining with acr...

  9. Validation of a flow cytometric acridine orange micronuclei methodology in rats.

    Science.gov (United States)

    Criswell, K A; Krishna, G; Zielinski, D; Urda, G A; Juneau, P; Bulera, S; Bleavins, M R

    2003-07-25

    Our laboratory has previously reported a flow cytometric acridine orange method for detection of micronucleus (MN) in the rat using cyclophosphamide as a test compound. To replace the manual method of scoring and satisfy Good Laboratory Practice (GLP) requirements, an extensive validation of the flow method was required. Therefore, manual scoring and flow cytometric determination of MN were compared using vincristine, chlorambucil, methotrexate, and doxorubicin compounds known to induce MN formation with various mechanisms of action. 1,2-Dimethylhydrazine (1,2-DH), a compound with negative or equivocal MN findings also was evaluated. The flow method consistently demonstrated dose- and time-dependent responses for MN production at all concentrations of vincristine, methotrexate, clorambucil, and doxorubicin. In contrast, manual scoring of slides failed to detect an increase in MN at the lowest doses of doxorubicin (1mg/kg) at 24 or 48 h, and methotrexate at 48 h, or any dose of methotrexate (50, 100, or 250 mg/kg) at 24h. Additionally, a dose-response for methotrexate at 48 h, and chlorambucil at 24 h were missed using manual scoring. For 1,2-DH, the flow method showed a low level (< 1.4-fold) increase in MN at all doses and times. In contrast, the manual method showed five-seven-fold increases at 24 h, but a < two-fold increase at 48 h in the highest dose only. These data may suggest that the flow method has a greater sensitivity and possibly accuracy than manual scoring. Significant decreases in polychromatic erythrocytes (PCE) were seen using both methods at approximately the same dose for all compounds. However, absolute flow cytometric PCE values were consistently higher than manual. Additional cytotoxicity parameters obtained by the flow method allowed a more complete assessment of cytotoxicity than PCE alone. Furthermore, data reported here combined with improved throughput, shortened data turnaround and reporting times, and possibly better precision due to

  10. A new acridine derivative as a highly selective fluoroionophore for Cu{sup 2+} in 100% aqueous solution

    Energy Technology Data Exchange (ETDEWEB)

    Wang, Yu, E-mail: wangyu1168@sxu.edu.cn [School of Chemistry and Chemical Engineering, Shanxi University, Taiyuan 030006 (China); Shi, LiLi; Sun, HuanHe Sun; Shang, ZhuoBin; Chao, JianBin [School of Chemistry and Chemical Engineering, Shanxi University, Taiyuan 030006 (China); Jin, WeiJun, E-mail: wjjin@bnu.edu.cn [College of Chemistry, Beijing Normal University, Beijing 100875 (China)

    2013-07-15

    A new acridine fluoroionophore containing two iminodiacetic acid ligands, Acridinyl Tetra Acid (ATA), was synthesized. The fluorescence sensing behavior of ATA toward metal ions was investigated in buffered aqueous media. The presence of Cu{sup 2+} resulted in significant quenching of the fluorescence emission from ATA, while other metal ions posed little interferences, if any. The fluorescence response was concentration-dependent and can be well described by the modified Stern–Volmer equation. A good linear relationship (R{sup 2}=0.9952) was observed up to 3.0×10{sup −6} mol L{sup −1} Cu{sup 2+} ions. The detection limit, calculated following the 3σ IUPAC criteria, was 1.24×10{sup −7} mol L{sup −1}. The presence of Cu{sup 2+} induces the formation of a 1:1 ligand/metal complex at neutral pH. -- Highlights: ► A new acridine fluoroionophore was synthesized for detection of Cu{sup 2+} in water. ► Iminodiacetic acid moiety was chosen as the ionophore. ► The fluoroionophore exhibits great fluorescence quenching effect for Cu{sup 2+}.

  11. In-depth validation of acridine orange staining for flow cytometric parasite and reticulocyte enumeration in an experimental model using Plasmodium berghei

    DEFF Research Database (Denmark)

    Hein-Kristensen, L; Wiese, L; Kurtzhals, J A L;

    2009-01-01

    Flow cytometry is potentially an effective method for counting malaria parasites, but inconsistent results have hampered its routine use in rodent models. A published two-channel method using acridine orange offers clear discrimination between the infected and uninfected erythrocytes. However...

  12. Evaluation of new iodinated acridine derivatives for targeted radionuclide therapy of melanoma using 125I, an Auger electron emitter

    International Nuclear Information System (INIS)

    The full text of the publication follows. The increasing incidence of melanoma and the lack of effective therapy on the disseminated form have led to an urgent need for new specific therapies. Several iodo-benzamides or analogs are known to possess specific affinity for melanoma tissue. New hetero-aromatic derivatives have been designed with a cytotoxic moiety and termed DNA intercalating agents. These compounds could be applied in targeted radionuclide therapy using 125I, Auger electrons emitter which gives high-energetic localized irradiation. Two iodinated acridine derivatives have been reported to present an in vivo kinetic profile conducive to application in targeted radionuclide therapy. The aim of the present study was to perform a preclinical evaluation of these compounds. The DNA intercalating property was confirmed for both compounds. After radiolabeling with 125I, the two compounds induced in vitro a significant radiotoxicity on B16F0 melanoma cells. The acridine compound, ICF01040, appeared more radio toxic than the acridone compound, ICF01035. While cellular uptake was similar for both compounds, SIMS analysis and in vitro protocol showed a stronger affinity for melanin with ICF01035, which was able to induce a predominant scavenging process in the melanosome and restrict access to the nucleus. Nevertheless, an important radiotoxicity was measured for the two compounds while the nuclear accumulation was low. Indeed, even if nuclear localization remains the main target sensitive to Auger electrons, the cell membrane remains sensitive to 125I decays. So, these compounds may induce secondary toxic effects of irradiation, such as membrane lipid damage. Conducted to current experiments are evaluate such hypothesis. Taken together, these results suggest that ICF01040 is a better candidate for application in targeted radionuclide therapy using 125I. The next step will be in vivo evaluation, where high tumoral vectorization gives promising perspectives

  13. Determination of ACC-induced cell-programmed death in roots of Vicia faba ssp. minor seedlings by acridine orange and ethidium bromide staining

    OpenAIRE

    Byczkowska, Anna; Kunikowska, Anita; Kaźmierczak, Andrzej

    2012-01-01

    Fluorescence staining with acridine orange (AO) and ethidium bromide (EB) showed that nuclei of cortex root cells of 1-aminocyclopropane-1-carboxylic acid (ACC)-treated Vicia faba ssp. minor seedlings differed in color. Measurement of resultant fluorescence intensity (RFI) showed that it increased when the color of nuclear chromatin was changed from green to red, indicating that EB moved to the nuclei via the cell membrane which lost its integrity and stained nuclei red. AO/EB staining showed...

  14. Synthesis of a new class of strongly fluorescent heterocyclic compounds: 3H-imidazo[4,5-a]acridine-11-carbonitriles

    Energy Technology Data Exchange (ETDEWEB)

    Sahraei, Robabeh [Department of Chemistry, Mashhad Branch, Islamic Azad University, Mashhad (Iran, Islamic Republic of); Pordel, Mehdi, E-mail: mehdipordel58@yahoo.com [Department of Chemistry, Mashhad Branch, Islamic Azad University, Mashhad (Iran, Islamic Republic of); Behmadi, Hossein; Razavi, Bahareh [Department of Chemistry, Mashhad Branch, Islamic Azad University, Mashhad (Iran, Islamic Republic of)

    2013-04-15

    The synthesis, spectral characterization and fluorescence studies of some novel substituted 8-chloro-3H-imidazo[4,5-a]acridine-11-carbonitriles are presented. The nucleophilic substitution of hydrogen in 5-nitrobenzimidazole derivatives occurs upon reaction with 2-(4-chlorophenyl) acetonitrile under basic conditions and proceeds with concomitant cyclisation. This sequence offers a one-pot synthesis of new fluorescent heterocyclic compounds, 8-chloro-3-alkyl-3H-imidazo[4,5-a]acridine-11-carbonitriles. The fluorescence of all compounds was intense and fluorescence quantum yields were very high (>0.85) for all of them. -- Highlights: ► A facile method to synthesis of new 3H-imidazo[4,5–a]acridine-11-carbonitriles is presented. ► The spectral and photophysical properties of these new compounds are investigated. ► The fluorescence of all compounds was intense and fluorescence quantum yields were very high for all of them.

  15. Low-dose Radiation-Induced Adaptive Response in Polychromatic Mice Erythrocyte as Measures by Acridine Orange Stained Micronucleus Assay

    International Nuclear Information System (INIS)

    The effect of conditioning pretreatment with 0.01Gy of gamma rays on micronucleated polychromatic erythrocyte (MN-PCE) induction by 2Gy of g-rays was determined in peripheral blood of C3H/He mice. The timing of their administration of challenge doses was 6 hr. The response was determined by scoring of Acridine orange due stained MN-PCEs. The results indicate that low dose gamma ray pretreatment does protect against MN-PCE induction by the challenge g-ray dose. Introduction: an adaptive response induced by low doses of ionizing radiation in vivo reported. Some research team reports that a reduction on MN-PCE of mice caused by the pretreatment was observed (1-4). However, there was variability in the amount of the response depending on the time and adaptive dose (3). This is important because the variation of MN-PCE frequency with time could lead to differences in the interpretation. In this study, differences in the biological effects within the priming dose ranges are discussed. (Author)

  16. Sensitive and selective turn-on fluorescence method for cetyltrimethylammonium bromide determination based on acridine orange-polystyrene sulfonate complex.

    Science.gov (United States)

    Li, Na; Hao, Xia; Kang, Bei Hua; Li, Nian Bing; Luo, Hong Qun

    2016-06-01

    This work proposed a rapid and novel fluorescence-sensing system using a complex of acridine orange (AO) and polystyrene sulfonate (PSS) to sensitively recognize and monitor cetyltrimethylammonium bromide (CTAB) in an aqueous medium. AO can interact with PSS and a complex is formed via electrostatic attraction and hydrophobic interaction. The fluorescence of AO is greatly quenched after the introduction of PSS. Upon its subsequent addition, CTAB can interact and form a complex with PSS because the electrostatic attraction between CTAB and PSS is much stronger than that between AO and PSS, which results in significant fluorescence recovery. Interestingly, the proposed method can be applied for the discrimination and detection of surfactants with different hydrocarbon chain lengths due to their different binding affinity toward PSS. The detection limit for CTAB is as low as 0.2 µg/mL and the linear range is from 0.5 to 3.5 µg/mL. Moreover, we applied the sensor to the successful detection of CTAB in water samples. Copyright © 2015 John Wiley & Sons, Ltd.

  17. Inhibition of RNA recruitment and replication of an RNA virus by acridine derivatives with known anti-prion activities.

    Directory of Open Access Journals (Sweden)

    Zsuzsanna Sasvari

    Full Text Available BACKGROUND: Small molecule inhibitors of RNA virus replication are potent antiviral drugs and useful to dissect selected steps in the replication process. To identify antiviral compounds against Tomato bushy stunt virus (TBSV, a model positive stranded RNA virus, we tested acridine derivatives, such as chlorpromazine (CPZ and quinacrine (QC, which are active against prion-based diseases. METHODOLOGY/PRINCIPAL FINDINGS: Here, we report that CPZ and QC compounds inhibited TBSV RNA accumulation in plants and in protoplasts. In vitro assays revealed that the inhibitory effects of these compounds were manifested at different steps of TBSV replication. QC was shown to have an effect on multiple steps, including: (i inhibition of the selective binding of the p33 replication protein to the viral RNA template, which is required for recruitment of viral RNA for replication; (ii reduction of minus-strand synthesis by the tombusvirus replicase; and (iii inhibition of translation of the uncapped TBSV genomic RNA. In contrast, CPZ was shown to inhibit the in vitro assembly of the TBSV replicase, likely due to binding of CPZ to intracellular membranes, which are important for RNA virus replication. CONCLUSION/SIGNIFICANCE: Since we found that CPZ was also an effective inhibitor of other plant viruses, including Tobacco mosaic virus and Turnip crinkle virus, it seems likely that CPZ has a broad range of antiviral activity. Thus, these inhibitors constitute effective tools to study similarities in replication strategies of various RNA viruses.

  18. Design and synthesis of novel anti-Alzheimer's agents: Acridine-chromenone and quinoline-chromenone hybrids.

    Science.gov (United States)

    Najafi, Zahra; Saeedi, Mina; Mahdavi, Mohammad; Sabourian, Reyhaneh; Khanavi, Mahnaz; Tehrani, Maliheh Barazandeh; Moghadam, Farshad Homayouni; Edraki, Najmeh; Karimpor-Razkenari, Elahe; Sharifzadeh, Mohammad; Foroumadi, Alireza; Shafiee, Abbas; Akbarzadeh, Tahmineh

    2016-08-01

    A novel series of acridine-chromenone and quinoline-chromenone hybrids were designed, synthesized, and evaluated as anti-Alzheimer's agents. All synthesized compounds were evaluated as cholinesterases (ChEs) inhibitors and among them, 7-(4-(6-chloro-2,3-dihydro-1H-cyclopenta[b]quinolin-9-ylamino)phenoxy)-4-methyl-2H-chromen-2-one (8e) exhibited the most potent anti-acetylcholinesterase (AChE) inhibitory activity (IC50=16.17μM) comparing with rivastigmine (IC50=11.07μM) as the reference drug. Also, compound 8e was assessed for its β-secretase (BACE1) inhibitory and neuroprotective activities which demonstrated satisfactory results. It should be noted that both kinetic study on the inhibition of AChE and molecular modeling revealed that compound 8e interacted simultaneously with both the catalytic active site (CAS) and peripheral anionic site (PAS) of AChE. PMID:27289559

  19. Low-Dose Radiation-Induced Adaptive Response in Polychromatic Mice Erythrocyte as Measured by Acridine Orange Stained Micronuleus Assay

    International Nuclear Information System (INIS)

    The effect of conditioning pretreatment with 0.01Gy of gamma rays on micronucleated polychromatic erythrocyte (MN-PCE) induction by 2Gy of g-rays was determined in peripheral blood of C3H/He mice. The timing of their administration of challenge doses was 6hr. The response was determined by scoring of Acridine orange dye stained MN-PCEs. The results indicate that low dose gamma ray pretreatment does protect against MN-PCE induction by the challenge g-ray dose. Introduction: An adaptive response induced by low doses of ionizing radiation in vivo reported. Some research team reports that a reduction on MN-PCE of mice caused by the pretreatment was observed [1- 4]. However, there was variability in the amount of the response depending on the time and adaptive dose [3]. This is important because the variation of MN-PCE frequency with time could lead to differences in the interpretation. In this study, differences in the biological effects within the priming dose ranges are discussed. Materials and Methods: Specific pathogen free 5-week-old C3H/He mice, purchased from Shizuoka Laboratory Center (Japan), were kept in clean and conventional environment. When 6 weeks old, the animal were whole body irradiated using irradiator of IBL-437 (137Cs, 0.8Gy/min). After various time intervals, the two groups were administrated to adaptation dose and challenge dose of 0.01Gy and 2Gy, respectively. For experiments, sham-irradiated, only adaptive and challenge dose irradiated groups were run concurrently. Smears were stained and scored using Acridine orange dye method [2]. Statistically significant differences in MN-PCE frequency were determined by comparing tie individual values at each group with the respective control values (challenge dose irradiated group) by using the paired ttest. Results and Discussions: Induced MN by the challenge dose (2Gy) after the pretreatment with 0.01Gy is low to the one induced by the challenge dose alone. In the present study, this estimation for the

  20. Preliminary biological evaluation of acridinic compounds for a targeted combined chemo and internal radionuclide therapy for melanoma

    International Nuclear Information System (INIS)

    The increasing incidence of melanoma and a lack of effective therapy on the disseminated form induces the development of selective tissue-targeted therapies. The aim of the present work was a targeting approach combining a bimodality therapy with the same compound exhibiting both chemo and internal radionuclide therapeutic properties. Benzamides are known to present a specific affinity for melanoma tissue. Former studies have shown that with aromatic and hetero-aromatic analogues of N-(2-diethylaminoethyl)- 4-iodo benzamide (B.Z.A.), the affinity for melanoma was maintained. In this context, new compounds have been designed and synthesized conjugating a cytotoxic hetero-aromatic moiety, an amino-alkyl amidic side chain for melanoma targeting and a radioiodine for internal radionuclide therapy. Acridinic derivatives known as cytotoxic DNA-intercalating agents have been chosen for this study. The cytotoxic activity of fifteen new compounds has been tested in vitro on a panel of cell lines and the I.C.50 values were determined. The three first selected compounds have been further evaluated: in vivo, on B 16 F0 melanoma bearing C 57 B.L.6 mice to determine the pharmacological kinetic and namely the tumoral affinity. Two compounds exhibited a high, specific and long lasting concentration in melanoma tumor giving them a kinetic profile favourable for an application to radionuclide therapy; in vitro, using the 'colony forming' test on melanoma cells, for a first approach of association of chemo toxicity and radiotoxicity. Assessed on the ability of cells to form colonies, the inhibition observed with the association for a same molecule of chemo toxic and radio toxic doses was quite exactly the sum of the two separate effects, a result providing a first validation of the radio chemotherapy concept; in vitro, by a preliminary determination of molecular mechanisms. Compared to parent compounds, results confirmed a maintain of DNA-intercalating properties. These first results

  1. 32P-postlabelling analysis of dibenz[a,j]acridine-DNA adducts in mice: identification of proximate metabolites.

    Science.gov (United States)

    Talaska, G; Roh, J; Schamer, M; Reilman, R; Xue, W; Warshawsky, D

    1995-03-30

    N-Heterocyclic polynuclear aromatics are widely-occurring environmental pollutants formed during the pyrolysis of nitrogen-containing organic chemicals. Dibenz[a,j]acridine (DBA), a member of this class, has been shown to be a skin carcinogen in mice. We undertook studies to determine the organ distribution of DBA-DNA adducts and to identify the DBA metabolites which lead to the formation of carcinogen-DNA adducts in vivo. DBA and its metabolites, trans-DBA-1,2-dihydrodiol (DBA-1,2-DHD) trans-DBA-3,4-dihydrodiol (DBA-3,4-DHD) and trans-DBA-5,6-dihydrodiol (DBA-5,6-DHD), were topically applied on mice. DNA was isolated using enzyme-solvent extraction methods, and analyzed for carcinogen-DNA adducts using 32P-postlabelling. In skin, DBA produced two distinct adducts (Adducts 1 and 2). The same two adducts were seen when DBA-3,4-DHD was applied. In addition, the total adduct level elicited by DBA-3,4-DHD was twice that of the parent compound. Two adducts (Adducts 3 and 4) were also seen in mouse skin when DBA-5,6-DHD was applied, but these differed chromatographically from adducts seen with DBA. However, when DBA-3,4-DHD was applied and analyzed using sensitive nuclease P1 32P-postlabelling, all four adducts could be detected. These results suggest that the major route of DBA activation to DNA-binding species in skin is through formation of DBA-3,4-DHD and subsequent metabolism of this compound to a bay-region diol-epoxide. However, we postulate that another activation pathway may proceed through a bis-dihydrodiol-epoxide.

  2. Synthesis of modified maghemite nanoparticles and its application for removal of Acridine Orange from aqueous solutions by using Box-Behnken design

    Energy Technology Data Exchange (ETDEWEB)

    Bagheban Shahri, Fatemeh; Niazi, Ali, E-mail: a-niazi@iau-arak.ac.ir

    2015-12-15

    In this study, sodium dodecyl sulfate-coated maghemite nanoparticles (SDS-coated γ-Fe{sub 2}O{sub 3} NPs), was used for removal of cationic dye Acridine Orange from water samples. The γ-Fe{sub 2}O{sub 3} NPs were synthesized by co-precipitation method and were characterized by scanning electron microscope (SEM) and vibrating sample magnetometer (VSM) to examine their size and magnetic moment. The adsorption experiments were performed using the batch system. The prepared magnetic adsorbent was well dispersed in water and easily separated magnetically from the medium after loaded with adsorbate. Four most important operating variables including initial pH of the solution, dosage of adsorbent, concentration of dye and contact time was studied and optimized by response surface methodology (RSM), involving Box-Behnken design matrix. Twenty-seven experiments were performed to investigate the effect of these parameters on removal of the dye. The results showed that initial pH of the solution was the most effective parameter in comparison with others. Also, experimental parameters were optimized and chose the best conditions by determination of effective factors. The optimized conditions for dye removal were at initial pH 5.1 0.8 g L{sup −1} of adsorbent, 30.0 mg L{sup −1} dye and 43 min adsorption time. The experimental data were analyzed by the Langmuir and Freundlich adsorption models. The maximum predicted adsorption capacities for Acridine Orange was 285.82 mg g{sup −1}. - Highlights: • Synthesis of maghemite as magnetic nanoparticle by co-precipitation. • Simple and fast removal of Acridine Orange by MMNPs. • Effect of parameters are optimized by Box-Behnken design.

  3. Preliminary biological evaluation of acridinic compounds for a targeted combined chemo and internal radionuclide therapy for melanoma

    Energy Technology Data Exchange (ETDEWEB)

    Gardette, M.; Papon, J.; Desbois, N.; Labarre, P.; Maisonial, A.; Maublant, J.; Madelmont, J.C.; Moins, N.; Chezal, J.M. [Centre Jean Perrin, Inserm-Universite d' Auvergne, 63 - Clermont Ferrand (France)

    2008-02-15

    The increasing incidence of melanoma and a lack of effective therapy on the disseminated form induces the development of selective tissue-targeted therapies. The aim of the present work was a targeting approach combining a bimodality therapy with the same compound exhibiting both chemo and internal radionuclide therapeutic properties. Benzamides are known to present a specific affinity for melanoma tissue. Former studies have shown that with aromatic and hetero-aromatic analogues of N-(2-diethylaminoethyl)- 4-iodo benzamide (B.Z.A.), the affinity for melanoma was maintained. In this context, new compounds have been designed and synthesized conjugating a cytotoxic hetero-aromatic moiety, an amino-alkyl amidic side chain for melanoma targeting and a radioiodine for internal radionuclide therapy. Acridinic derivatives known as cytotoxic DNA-intercalating agents have been chosen for this study. The cytotoxic activity of fifteen new compounds has been tested in vitro on a panel of cell lines and the I.C.50 values were determined. The three first selected compounds have been further evaluated: in vivo, on B 16 F0 melanoma bearing C 57 B.L.6 mice to determine the pharmacological kinetic and namely the tumoral affinity. Two compounds exhibited a high, specific and long lasting concentration in melanoma tumor giving them a kinetic profile favourable for an application to radionuclide therapy; in vitro, using the 'colony forming' test on melanoma cells, for a first approach of association of chemo toxicity and radiotoxicity. Assessed on the ability of cells to form colonies, the inhibition observed with the association for a same molecule of chemo toxic and radio toxic doses was quite exactly the sum of the two separate effects, a result providing a first validation of the radio chemotherapy concept; in vitro, by a preliminary determination of molecular mechanisms. Compared to parent compounds, results confirmed a maintain of DNA-intercalating properties. These

  4. Microwave assisted synthesis of novel acridine-acetazolamide conjugates and investigation of their inhibition effects on human carbonic anhydrase isoforms hCA I, II, IV and VII.

    Science.gov (United States)

    Ulus, Ramazan; Aday, Burak; Tanç, Muhammet; Supuran, Claudiu T; Kaya, Muharrem

    2016-08-15

    4-Amino-N-(5-sulfamoyl-1,3,4-thiadiazol-2-yl)benzamide was condensed with cyclic-1,3-diketones (dimedone and cyclohexane-1,3-dione) and aromatic aldehydes under microwave irradiation, leading to a series of acridine-acetazolamide conjugates. The new compounds were investigated as inhibitors of carbonic anhydrases (CA, EC 4.2.1.1), and more precisely cytosolic isoforms hCA I, II, VII and membrane-bound one hCA IV. All investigated isoforms were inhibited in low micromolar and nanomolar range by the new compounds. hCA IV and VII were inhibited with KIs in the range of 29.7-708.8nM (hCA IV), and of 1.3-90.7nM (hCA VII). For hCA I and II the KIs were in the range of 6.7-335.2nM (hCA I) and of 0.5-55.4nM (hCA II). The structure-activity relationships (SAR) for the inhibition of these isoforms with the acridine-acetazolamide conjugates reported here were delineated. PMID:27298005

  5. Synthesis of modified maghemite nanoparticles and its application for removal of Acridine Orange from aqueous solutions by using Box-Behnken design

    Science.gov (United States)

    Bagheban Shahri, Fatemeh; Niazi, Ali

    2015-12-01

    In this study, sodium dodecyl sulfate-coated maghemite nanoparticles (SDS-coated γ-Fe2O3 NPs), was used for removal of cationic dye Acridine Orange from water samples. The γ-Fe2O3 NPs were synthesized by co-precipitation method and were characterized by scanning electron microscope (SEM) and vibrating sample magnetometer (VSM) to examine their size and magnetic moment. The adsorption experiments were performed using the batch system. The prepared magnetic adsorbent was well dispersed in water and easily separated magnetically from the medium after loaded with adsorbate. Four most important operating variables including initial pH of the solution, dosage of adsorbent, concentration of dye and contact time was studied and optimized by response surface methodology (RSM), involving Box-Behnken design matrix. Twenty-seven experiments were performed to investigate the effect of these parameters on removal of the dye. The results showed that initial pH of the solution was the most effective parameter in comparison with others. Also, experimental parameters were optimized and chose the best conditions by determination of effective factors. The optimized conditions for dye removal were at initial pH 5.1 0.8 g L-1 of adsorbent, 30.0 mg L-1 dye and 43 min adsorption time. The experimental data were analyzed by the Langmuir and Freundlich adsorption models. The maximum predicted adsorption capacities for Acridine Orange was 285.82 mg g-1.

  6. Pd-Catalyzed Intramolecular Heck Reaction, C(sp(2))-H Activation, 1,4-Pd Migration, and Aminopalladation: Chemoselective Synthesis of Dihydroindeno[1,2,3-kl]acridines and 3-Arylindoles.

    Science.gov (United States)

    Gu, Zheng-Yang; Liu, Cheng-Guo; Wang, Shun-Yi; Ji, Shun-Jun

    2016-05-20

    Palladium-catalyzed intramolecular Heck reaction and aminopalladation of N-(2-(1-phenylvinyl)phenyl)aniline for the efficient synthesis of dihydroindeno[1,2,3-kl]acridines and 3-arylindoles via tuning of the phosphine ligands and solvents under two optimized conditions are reported. The reaction follows a 1,4-Pd migration, aminopalladation, C(sp(2))-H activation, as well as five- and six-membered-ring fusion to form different products. The dihydroindeno[1,2,3-kl]acridine derivatives showed higher triplet energy (ET) levels than common blue phosphorescent dopant and may serve as good host candidates for blue triplet emitters. PMID:27137482

  7. Associated electron and proton transfer between Acridine and Triethylamine in AOT reverse micelles probed by laser flash photolysis with magnetic field

    Science.gov (United States)

    Sarangi, Manas Kumar; Basu, Samita

    2011-04-01

    Laser flash photolysis with magnetic field (MF ˜0.08 T) has been used to study interaction between Acridine (Acr) and Triethylamine (TEA) in reverse micelles with w0 = 2.5-40. Dynamic protonation equilibrium exists between 3Acr and 3AcrH +. The intermediates indicate excited-state proton transfer (PT) between 3AcrH + and TEA. However, application of MF highlights the formation of geminate radical ion pairs (RIPs) with triplet spin-correlation, a signature of latent photoinduced electron transfer between 3AcrH + and TEA co-exists with PT. Magnetic field effect (MFE) is prominent for smaller w0 showing importance of optimum separation between RIP to maximize MFE, whereas PT remains unaltered.

  8. Design, Synthesis, Fluorescence Properties and Antibacterial Activities of New 8-Chloro-3-Alkyl-3H-Pyrazolo[4,3-a]acridine-11-Carbonitriles

    Energy Technology Data Exchange (ETDEWEB)

    Rahmani, Zeynab; Pordel, Mehdi; Davoodnia, Abolghasem [Islamic Azad Univ., Mashhad (Iran, Islamic Republic of)

    2014-02-15

    The treatment of alkylated nitro derivatives of indazole with 2-(4-chlorophenyl)acetonitrile under basic conditions gave the new 8-chloro-3-alkyl-3H-pyrazolo[4,3-a]acridine-11-carbonitriles via the nucleophilic substitution of hydrogen which proceeds at room temperature with concomitant cyclisation in fairly good yields. The structures of all newly synthesized compounds were confirmed by IR, {sup 1}H NMR, {sup 13}C NMR and mass spectral data. Fluorescence experimental results of all newly synthesized compounds revealed remarkable photoluminescence properties and strong green fluorescence properties. Also, the new compounds exhibited potent antibacterial activity and their antibacterial activity (MIC) against Gram positive (Staphylococcuse aureus methicillin resistant S. aureus and Bacillus subtilis) and negative bacterial (Pseudomonas aeruginosa and Escherichia coli) species were determined.

  9. Novel acridine-based N-acyl-homoserine lactone analogs induce endoreduplication in the human oral squamous carcinoma cell line SAS

    International Nuclear Information System (INIS)

    The cytotoxicity of novel acridine-based N-acyl-homoserine lactone (AHL) analogs was investigated on the human oral squamous carcinoma cell line SAS. One analog induced G2/M phase arrest at 5.3-10.6 μM and induced polyploidy at a higher dose (21.2 μM). Importantly, treatment of SAS cells with a combination of the AHL analog and the Jun N-terminal kinase (JNK) inhibitor, SP600125, prevented mitosis and induced polyploidy. The AHL analog synergized with X-irradiation to inhibit clonogenic survival of SAS cells; however, its radiosensitizing effects were relative to not X-irradiation-induced apoptosis but mitotic failure following enhanced expression of Aurora A and B. These results suggest that the active AHL analog showed growth-suppressive and radiosensitizing effects, which involve polyploidy followed by G2/M accumulation and atypical cell death in the SAS cell line. (author)

  10. Blood culture gram stain, acridine orange stain and direct sensitivity-based antimicrobial therapy of bloodstream infection in patients with trauma

    Directory of Open Access Journals (Sweden)

    Behera B

    2010-01-01

    Full Text Available Purpose: The purpose of this study was to ascertain if the simple practice of Gram stain, acridine orange stain and direct sensitivity determination of positive blood culture bottles could be used to guide early and appropriate treatment in trauma patients with clinical suspicion of sepsis. The study also aimed to evaluate the error in interpreting antimicrobial sensitivity by direct method when compared to standard method and find out if specific antibiotic-organism combination had more discrepancies. Findings from consecutive episodes of blood stream infection at an Apex Trauma centre over a 12-month period are summarized. Materials and Methods: A total of 509 consecutive positive blood cultures were subjected to Gram staining. AO staining was done in BacT/ALERT-positive Gram-stain negative blood cultures. Direct sensitivity was performed from 369 blood culture broths, showing single type of growth in Gram and acridine orange staining. Results of direct sensitivity were compared to conventional sensitivity for errors. Results: No ′very major′ discrepancy was found in this study. About 5.2 and 1.8% minor error rates were noted in gram-positive and gram-negative bacteria, respectively, while comparing the two methods. Most of the discrepancies in gram-negative bacteria were noted in β lactam - β lactamase inhibitor combinations. Direct sensitivity testing was not reliable for reporting of methicillin and vancomycin resistance in Staphylococci. Conclusions: Gram stain result together with direct sensitivity testing is required for optimizing initial antimicrobial therapy in trauma patients with clinical suspicion of sepsis. Gram staining and AO staining proved particularly helpful in the early detection of candidaemia.

  11. Supra­molecular inter­actions in a 1:1 co-crystal of acridine and 3-chloro­thio­phene-2-carb­oxy­lic acid

    OpenAIRE

    Prajina, Olakkandiyil; Thomas Muthiah, Packianathan; Perdih, Franc

    2016-01-01

    In the title co-crystal, C5H3ClO2S·C13H9N, the components inter­act with each other via an O—H⋯N hydrogen bond. Acridine–acridine stacking, thio­phene–thio­phene stacking and acridine–thio­phene C—H⋯π inter­actions also occur in the crystal.

  12. 吖啶橙指示荧光分析法测定双酚A%Spectrofluorimetric Determination of Bisphenol A with Acridine Orange as Indicator

    Institute of Scientific and Technical Information of China (English)

    杜凌云; 包玉红; 王术皓

    2011-01-01

    A new spectrofluorimetric method has been developed for the determination of bisphenol A based on the inhibitory effect of bisphenol A on redox reaction between hydroxyl radical and acridine orange in acidic media. This method is simple, fast, and its linear range is 1. 0 to 100 ng/mL with the detection limit of 0. 21 ng/mL. The proposed method was applied to the determination of bisphenol A in plastic bag with satisfactory results.%基于在酸性介质中,双酚A对羟自由基与吖啶橙的氧化还原反应的阻抑作用,建立了测定双酚A的荧光分析新方法.该方法简单,快速,线性范围为1.0~100 ng/mL,检出限为0.21 ng/mL.将其用于塑料制品中双酚A的测定,结果满意.

  13. Radioadaptive response in human B- and CD8{sup +} T-lymphocytes as measured by the acridine orange stained micronuclei technique

    Energy Technology Data Exchange (ETDEWEB)

    Kim, H.S.; Choi, J.M.; Yang, K.H.; Kim, C.S.; Lim, Y.K.; Kim, C.S. [Korea Hydro and Nuclear Power Corporation, Radiation Health Research Institute, Seoul (Korea); Woon, J.H. [National Veterinary Research and Quarantine Service, Anyang (Korea)

    2003-07-01

    To investigate (1) the radiosensitive of B- versus T- lymphocytes and (2) the possible application of their sensitivity for adaptive response after treating with an adapting plus a challenge dose. In the present experiments, micronucleus analysis was performed in B- and CD8{sup +} matured T-lymphocytes of eight healthy volunteers exposed to {gamma}-rays. The number of radio-induced micronuclei was significantly higher in B-lymphocytes compared to T-lymphocytes in the dose range from 10 to 100cGy. To investigate adaptive response, whole blood samples were irradiated in vitro with a pretreatment dose of 1cGy {sup 137}Cs {gamma}-irradiation. Six hours after their initiation, groups of cultures were subsequently exposed to a challenge dose of 100cGy {gamma}-irradiation. Following stimulation with PHA and PWM for T- and B-lymphocyte cultivation, lymphocytes were fixed at 72 hours and stained with acridine orange dye. B-lymphocytes exhibited a greater induction of adaptive response than those of CD8{sup +} matured T-lymphocytes, and when pretreated with 1cGy significantly fewer micronuclei induced by the challenge dose of 100cGy {gamma}-irradiation. The results suggest that the lower dose pretreatments are able to induce a significantly higher adaptive response in human B-lymphocytes, and this adaptive response may result from the DNA repair mechanism, which may lead to less residual damage. (author)

  14. A Novel Fluorescence Probe 9-(4-(1,2-diamine)benzene-N1-phenyl)acridine for Nitric Oxide Determination

    Institute of Scientific and Technical Information of China (English)

    DING Liyun; YUAN Fang; HUANG Lanfen; HUANG Jun; LIU Xiaofang; LIANG Bing

    2014-01-01

    A novel fluorescent probe 9-(4-(1,2-diamine)benzene-N1-phenyl)acridine (DABPA) was synthesized for the detection of nitric oxide (NO) and characterized by IR, 1H-NMR and EI-MS spectroscopy. Based on a photoelectron transfer mechanism, the fluorescence intensities of DABPA were investigated with the different concentrations of NO. Under the optimal experimental conditions, the fluorescence intensity of DABPA had a good linear relationship (R2=0.9977) with NO concentration in the range from 1×10-7 to 1.5×10-6 mol/L with a detection limit of 1×10-8 mol/L. The cytotoxicity induced by DABPA was evaluated by the MTT (3-(4,5-dimethylthiazol-2-yl)-2,5diphenyl tetrazolium bromide) assay for biological application. Furthermore, the probe DABPA had also been successfully applied to real-time image NO produced in PC12 cells in the presence of L-arginine.

  15. Microbial quality of lamb carcasses during processing and the acridine orange direct count technique (a modified DEFT) for rapid enumeration of total viable counts.

    Science.gov (United States)

    Sierra, M L; Sheridan, J J; McGuire, L

    1997-04-29

    This study was designed to set up a hazard analysis and critical control points (HACCP) system for sheep slaughtering operations at four different plants in Ireland and to determine the differences between plants in terms of microbial contamination. A single carcass area, the abdomen, was examined by swabbing and a microbiological profile was determined at different stages along the slaughter line. The level of contamination was assessed from the total bacteria counts, Enterobacteriaceae and Listeria spp. For the total counts, a modified direct epifluorescent filter technique (acridine orange direct count technique (AODC)) was developed and tested. No significant differences were found among plants in the levels of bacterial contamination. This was observed for all groups of organisms. The rapid direct technique (AODC) was found to be very successful. A correlation coefficient of 0.87 was obtained for this method and the standard plate count. Each test could be carried out in about 10-15 min and could be used to predict the standard plate count.

  16. Application of poly(acridine orange) and graphene modified carbon/ionic liquid paste electrode for the sensitive electrochemical detection of rutin

    International Nuclear Information System (INIS)

    A carbon/ionic liquid paste electrode (CILPE) prepared by 1-hexylpyridinium hexafluorophosphate as the binder was used as the substrate electrode. A layer of graphene oxide (GO) film was cast on CILPE surface (GO/CILPE) and the electropolymerization of acridine orange (AO) on electrode was further realized by cyclic voltammetry in the potential range from −1.40 V to 1.40 V, which could simultaneously reduce GO to graphene (GR) electrochemically. The fabricated PAO-GR/CILPE exhibited good electrochemical performances with higher conductivity and lower electron transfer resistance. Electrochemical behaviors of rutin were further investigated on the modified electrode in 0.1 mol/L pH 2.0 phosphate buffer solution by cyclic voltammetry with a pair of well-defined redox peaks appeared. The peak-to-peak separation (ΔEp) was calculated as 0.076 V, which proved a fast quasi-reversible electron transfer process and the electrochemical parameters of rutin on PAO-GR/CILPE were calculated. Under the optimal conditions, the linear relationship between the oxidation peak current of rutin and its concentration was obtained in the range from 0.03 to 800.0 μmol/L with the detection limit as 8.33 nmol/L (3σ). The PAO-GR/CILPE showed good selectivity, stability and reproducibility, which was further applied to detect rutin tablet samples with satisfactory results

  17. Spectrophotometric determination of low levels arsenic species in beverages after ion-pairing vortex-assisted cloud-point extraction with acridine red.

    Science.gov (United States)

    Altunay, Nail; Gürkan, Ramazan; Kır, Ufuk

    2016-01-01

    A new, low-cost, micellar-sensitive and selective spectrophotometric method was developed for the determination of inorganic arsenic (As) species in beverage samples. Vortex-assisted cloud-point extraction (VA-CPE) was used for the efficient pre-concentration of As(V) in the selected samples. The method is based on selective and sensitive ion-pairing of As(V) with acridine red (ARH(+)) in the presence of pyrogallol and sequential extraction into the micellar phase of Triton X-45 at pH 6.0. Under the optimised conditions, the calibration curve was highly linear in the range of 0.8-280 µg l(-1) for As(V). The limits of detection and quantification of the method were 0.25 and 0.83 µg l(-1), respectively. The method was successfully applied to the determination of trace As in the pre-treated and digested samples under microwave and ultrasonic power. As(V) and total As levels in the samples were spectrophotometrically determined after pre-concentration with VA-CPE at 494 nm before and after oxidation with acidic KMnO4. The As(III) levels were calculated from the difference between As(V) and total As levels. The accuracy of the method was demonstrated by analysis of two certified reference materials (CRMs) where the measured values for As were statistically within the 95% confidence limit for the certified values.

  18. Layer-by-layer films and colloidal dispersions of graphene oxide nanosheets for efficient control of the fluorescence and aggregation properties of the cationic dye acridine orange

    Science.gov (United States)

    Hansda, Chaitali; Chakraborty, Utsav; Hussain, Syed Arshad; Bhattacharjee, Debajyoti; Paul, Pabitra Kumar

    2016-03-01

    Chemically derived graphene oxide (GO) nanosheets have received great deal of interest for technological application such as optoelectronic and biosensors. Aqueous dispersions of GO become an efficient template to induce the association of cationic dye namely Acridine Orange (AO). Interactions of AO with colloidal GO was governed by both electrostatic and π-π stacking cooperative interactions. The type of dye aggregations was found to depend on the concentration of GO in the mixed ensemble. Spectroscopic calculations revealed the formation of both H and J-type dimers, but H-type aggregations were predominant. Preparation of layer-by-layer (LbL) electrostatic self-assembled films of AO and GO onto poly (allylamine hydrochloride) (PAH) coated quartz substrate is also reported in this article. UV-Vis absorption, steady state and time resolve fluorescence and Raman spectroscopic techniques have been employed to explore the detail photophysical properties of pure AO, AO/GO mixed solution and AO/GO LbL films. Scanning electron microscopy was also used for visual evidence of the synthesized nanodimensional GO sheets. The fluorescence quenching of AO in the presence of GO in aqueous solution was due to the interfacial photoinduced electron transfer (PET) from photoexcited AO to GO i.e. GO acts as an efficient quenching agent for the fluorescence emission of AO. The quenching is found to be static in nature. Raman spectroscopic results also confirmed the interaction of AO with GO and the electron transfer. The formation of AO/GO complex via very fast excited state electron transfer mechanism may be proposed as to prepare GO-based fluorescence sensor for biomolecular detection without direct labeling the biomolecules by fluorescent probe.

  19. Determination of ACC-induced cell-programmed death in roots of Vicia faba ssp. minor seedlings by acridine orange and ethidium bromide staining.

    Science.gov (United States)

    Byczkowska, Anna; Kunikowska, Anita; Kaźmierczak, Andrzej

    2013-02-01

    Fluorescence staining with acridine orange (AO) and ethidium bromide (EB) showed that nuclei of cortex root cells of 1-aminocyclopropane-1-carboxylic acid (ACC)-treated Vicia faba ssp. minor seedlings differed in color. Measurement of resultant fluorescence intensity (RFI) showed that it increased when the color of nuclear chromatin was changed from green to red, indicating that EB moved to the nuclei via the cell membrane which lost its integrity and stained nuclei red. AO/EB staining showed that changes in color of the nuclear chromatin were accompanied by DNA condensation, nuclei fragmentation, and chromatin degradation which were also shown after 4,6-diamidino-2-phenylindol staining. These results indicate that ACC induced programmed cell death. The increasing values of RFI together with the corresponding morphological changes of nuclear chromatin were the basis to prepare the standard curve; cells with green unchanged nuclear chromatin were alive while those with dark orange and bright red nuclei were dead. The cells with nuclei with green-yellow, yellow-orange, and bright orange chromatin with or without their condensation and fragmentation chromatin were dying. The prepared curve has became the basis to draw up the digital method for detection and determination of the number of living, dying, and dead cells in an in planta system and revealed that ACC induced death in about 20% of root cortex cells. This process was accompanied by increase in ion leakage, shortening of cells and whole roots, as well as by increase in weight and width of the apical part of roots and appearance of few aerenchymatic spaces while not by internucleosomal DNA degradation. PMID:22350735

  20. Microwave assisted synthesis of novel Hantzsch 1,4-dihydropyridines, acridine-l,8-diones and polyhydroquinolines bearing the tetrazolo [1,5-a]quinoline moiety and their antimicrobial activity assess

    Institute of Scientific and Technical Information of China (English)

    Niraj K. Ladani; Divyesh C. Mungra; Manish P. Patel; Ranjan G. Patel

    2011-01-01

    Microwave assisted efficient Hantzsch reaction via four-component coupling reactions of tetrazolo [l,5-a]quinoline-4-carbal-dehyde, dimedone/cyclohexane-l,3-dione, ethyl/methyl acetoacetate and ammonium acetate was described as the preparation of tetrazolo [l,5-a]quinoline based 1,4-dihydropyridines, acridine-l,8-diones and polyhydroquinolines. The process presented here is simple, rapid, environmentally welcoming and high yielding. All the derivatives were subjected to an in vitro antimicrobial screening against a representative panel of bacteria and fungi and results worth further investigations.

  1. Electrochemical DNA biosensor for the detection of Trichoderma harzianum based on a gold electrode modified with a composite membrane made from an ionic liquid, ZnO nanoparticles and chitosan, and by using acridine orange as a redox indicator

    International Nuclear Information System (INIS)

    An electrochemical DNA biosensor was developed that is based on a gold electrode modified with a nanocomposite membrane made from an ionic liquid, ZnO nanoparticles and chitosan. A single-stranded DNA probe was immobilized on this electrode. Acridine orange was used as the hybridization probe for monitoring the hybridization of the target DNA. The biosensor was capable of detecting target DNA in the concentration range from 1.0 x 10-14 to 1.8 x 10-4 mol L-1, with a detection limit of 1.0 x 10-15 mol L-1. The approach towards constructing a DNA biosensor allows studies on the hybridization even with crude DNA fragments and also to analyze sample obtained from real samples. The results show that the DNA biosensor has the potential for sensitive detection of a specific sequence of the Trichoderma harzianum gene and provides a quick, sensitive and convenient method for the study of microorganisms. (author)

  2. Antigenotoxic and Apoptotic Activity of Green Tea Polyphenol Extracts on Hexavalent Chromium-Induced DNA Damage in Peripheral Blood of CD-1 Mice: Analysis with Differential Acridine Orange/Ethidium Bromide Staining

    Directory of Open Access Journals (Sweden)

    María del Carmen García-Rodríguez

    2013-01-01

    Full Text Available This study was conducted to investigate the modulating effects of green tea polyphenols on genotoxic damage and apoptotic activity induced by hexavalent chromium [Cr (VI] in CD-1 mice. Animals were divided into the following groups: (i injected with vehicle; (ii treated with green tea polyphenols (30 mg/kg via gavage; (iii injected with CrO3 (20 mg/kg intraperitoneally; (iv treated with green tea polyphenols in addition to CrO3. Genotoxic damage was evaluated by examining micronucleated polychromatic erythrocytes (MN-PCEs obtained from peripheral blood at 0, 24, 48, and 72 h after treatment. Induction of apoptosis and cell viability were assessed by differential acridine orange/ethidium bromide (AO/EB staining. Treatment of green tea polyphenols led to no significant changes in the MN-PCEs. However, CrO3 treatment significantly increased MN-PCEs at 24 and 48 h after injection. Green tea polyphenols treatment prior to CrO3 injection led to a decrease in MN-PCEs compared to the group treated with CrO3 only. The average of apoptotic cells was increased at 48 h after treatment compared to control mice, suggesting that apoptosis could contribute to eliminate the DNA damaged cells induced by Cr (VI. Our findings support the proposed protective effects of green tea polyphenols against the genotoxic damage induced by Cr (VI.

  3. 中性红、吖啶橙及PE标记的LAMP-2抗体在B16F10细胞溶酶体检测中的应用与比较%Application and comparison of neutral red, acridine orange, or PE labeled LAMP-2 antibodies in the detection of lysosomes in B16F10 cells

    Institute of Scientific and Technical Information of China (English)

    翟晓峰; 施文; 李国兴; 孙永强; 赵文静; 钱红燕; 李静; 陈橼; 何向锋

    2012-01-01

    We aimed to investigate the application and value of neutral red, acridine orange, or PE labeled anti-LAMP-2 antibodies in the detection of lysosomes in B16F10 cells. Firstly, we labeled the lysosomes of B16F10 cells with neutral red, acridine orange, and PE labeled anti-LAMP-2 antibodies respectively and detect the labeled lysosomes by optical and fluorescent microscopy. The results showed that the distribution and numbers of lysosomes in B16F10 cells could be clearly observed in optical microscope through the staining of neutral red, and the cytoplasm and lysosome could be stained in green and red respectively by acridine orange, but the quenching of red fluorescence in lysosome was so fast that the detection window was too narrow. The location and numbers of lysosomes in B16F10 cells could be clearly revealed with red fluorescence after the application of PE-labeled anti-LAMP-2 antibody, and the relative location of lysosomes and nucleus could be presented directly along with DAPI staining. In conclusion, the neutral red, acridine orange and PE labeled LAMP-2 antibody staining have their own advantage and characteristics in the detection of lysosomes. The choice of the effective lysosomes detection method should be based on the aim of study.%目的 探讨中性红、吖啶橙及PE标记的LAMP-2抗体在小鼠黑色素瘤B16F10细胞溶酶体检测中的应用与价值.方法 分别用中性红、吖啶橙和PE标记的LAMP-2抗体标记B16F10细胞溶酶体,通过光学和荧光显微镜进行检测.结果 中性红染色法能够在光镜下清晰显示溶酶体在细胞中的分布与数量,并能够反应溶酶体的功能;吖啶橙能够同时将细胞质和溶酶体分别用绿色和红色荧光标示出来,但溶酶体的红色荧光淬灭很快,观测窗口较窄;PE标记的LAMP-2抗体能够将溶酶体在细胞内的位置和数量清晰以红色荧光呈现出来,配合DAPI染色,可以直观显示溶酶体同细胞核

  4. Study on the Interaction between CdTe Quantum Dot-Acridine Orange-Calf Thymus DNA by Fluorescence Reversible Control%荧光可逆调控研究CdTe量子点-吖啶橙-小牛胸腺DNA的相互作用及分析应用

    Institute of Scientific and Technical Information of China (English)

    龚会平; 刘绍璞; 殷鹏飞; 闫曙光; 范小青; 何佑秋

    2011-01-01

    水相合成了谷胱甘肽(GSH)修饰的CdTe量子点(QDs).在PH=7.4的Tris-HCl缓冲溶液中,吖啶橙(AO)通过静电引力吸附到GSH-CdTe QDs的表面,与GSH-CdTe QDs形成了基态复合物,导致GSH-CdTe QDs的荧光猝灭.在GSH-CdTe QDs-AO体系中加入小牛胸腺DNA(ctDNA),ctDNA诱导AO从GSH-CdTe QDs表面脱落嵌入其双螺旋结构中,导致GSH-CdTe QDs的荧光恢复.根据GSH-CdTe QDs荧光的猝灭和恢复,实现了量子点荧光的可逆调控.ctDNA引起GSH-CdTe QDs-AO体系荧光恢复强度与ctDNA浓度成良好的线性关系,检出限为0.13 ng mL-1,据此提出了简便快捷、准确、高灵敏测定ctDNA的新方法.还结合共振瑞利散射(RRS)光谱、吸收光谱和原子力显微镜照片研究了GSH-CdTe QDs-AO-ctDNA三者之间的相互作用,对相互作用机理进行了讨论并提出了相应的作用模型.%Glutathione(GSH)-capped CdTe quantum dots(GSH-CdTe QDs) were synthesized in aqueous solution.In pH 7.4 Tris-HCl buffer medium,acridine orange(AO) was adsorbed to the surfaces of GSH-CdTe QDs via electrostatic attraction and formed ground state complex,which resulted in the quenching of the fluorescence of GSH-CdTe QDs.Adding ctDNA to GSH-CdTe QDs-AO system leaded to the fluorescence intensity of GSH-CdTe QDs recover,which can be explained by that the addition of ctDNA to the system induced AO to dissociate from the surface of GSH-CdTe QDs and embed into its double helix structure.According to the fluorescence quencher and restoration for GSH-CdTe QDs,fluorescence reversible control of QDs was realized.The fluorescence intensity change of GSH-CdTe QDs-AO system aroused by the addition of ctDNA was proportional to the ctDNA concentration in a certain range,and its detection limit was 0.13 ngomL-1.Based on it,the simple,rapid,accurate and sensitive methods had been proposed to determine ctDNA.The interaction of GSH-CdTe QDs-AO-ctDNA was studied by resonance Rayleigh scattering

  5. Structure of acridines and their effect on RNA synthesis in vitro.

    Science.gov (United States)

    Szmigiero, L; Slaska, K; Ciesielska, E; Jaros-Kamińska, B; Gniazdowski, M

    1977-01-01

    1. Ledakrin (C-283), a 1-nitro-9-aminopropylacridine derivative, inhibits RNA synthesis in vitro if the complex with DNA is formed in the presence of thiol. 2. Using several analogues of Ledakrin, it has been found that the 1-nitro group is essential for enhancement of the inhibition by thiol; the length of 9-aminoalkyl side chain also plays a role in the reaction between DNA and the dye. 3. It is suggested that the low inhibitory effect of Ledakrin in the absence of thiol compounds is due to a steric hindrance between neighbouring 1-nitro and 9-aminoalkyl groups. This hypothesis has been confirmed by assaying inhibition of RNA synthesis by several analogues of Ledakrin. PMID:868436

  6. Targeted photocytotoxicity by copper(II) complexes having vitamin B6 and photoactive acridine moieties.

    Science.gov (United States)

    Mukherjee, Nandini; Podder, Santosh; Banerjee, Samya; Majumdar, Shamik; Nandi, Dipankar; Chakravarty, Akhil R

    2016-10-21

    Copper(II) pyridoxal Schiff base complexes [Cu(L(1)/L(2))(B)]ClO4 (1-4), where HL(1) is 4-(((2-(1H-imidazol-4-yl)ethyl)imino)methyl)-5-(hydroxymethyl)-2-methylpyridin-3-ol (in 1 and 2), HL(2) is 2-(((2-(1H-imidazol-4-yl)ethyl)imino)methyl)phenol (in 3, 4), B is 11-(9-acridinyl)dipyrido[3,2-a:2',3'-c]phenazine (acdppz in 1 and 3), dipyrido[3,2-a:2',3'-c]phenazine (in 2) and 1,10-phenanthroline (in 4), were synthesized, characterized and their photocytotoxicity in visible light, intracellular localization, cellular uptake and DNA photocleavage activity were studied. Complex 4 was characterized by X-ray crystallography. Complexes 1 and 3 having acdppz as photosensitizer showed significant photocytotoxicity in visible light in HeLa and MCF7 cells giving IC50 value of <0.6 μM, while being relatively non-toxic in dark. The complexes were non-toxic to non-tumorigenic HPL1D cells both in light and dark conditions. Complex 1 showed significant localization in the cytoplasm of HeLa cells within 4 h of treatment, as evidenced from confocal microscopy. DCFDA assay on 1 suggested generation of intracellular reactive oxygen species in HeLa cells upon photo-exposure. Importantly, Annexin-V-FITC/PI assay indicated photo-induced apoptotic cell death. PMID:27423638

  7. Evaluation of two (125)I-radiolabeled acridine derivatives for Auger-electron radionuclide therapy of melanoma.

    Science.gov (United States)

    Gardette, Maryline; Viallard, Claire; Paillas, Salomé; Guerquin-Kern, Jean-Luc; Papon, Janine; Moins, Nicole; Labarre, Pierre; Desbois, Nicolas; Wong-Wah-Chung, Pascal; Palle, Sabine; Wu, Ting-Di; Pouget, Jean-Pierre; Miot-Noirault, Elisabeth; Chezal, Jean-Michel; Degoul, Francoise

    2014-08-01

    We previously selected two melanin-targeting radioligands [(125)I]ICF01035 and [(125)I]ICF01040 for melanoma-targeted (125)I radionuclide therapy according to their pharmacological profile in mice bearing B16F0 tumors. Here we demonstrate in vitro that these compounds present different radiotoxicities in relation to melanin and acidic vesicle contents in B16F0, B16F0 PTU and A375 cell lines. ICF01035 is effectively observed in nuclei of achromic (A375) melanoma or in melanosomes of melanized melanoma (B16F0), while ICF01040 stays in cytoplasmic vesicles in both cells. [(125)I]ICF01035 induced a similar survival fraction (A50) in all cell lines and led to a significant decrease in S-phase cells in amelanotic cell lines. [(125)I]ICF01040 induced a higher A50 in B16 cell lines compared to [(125)I]ICF01035 ones. [(125)I]ICF01040 induced a G2/M blockade in both A375 and B16F0 PTU, associated with its presence in cytoplasmic acidic vesicles. These results suggest that the radiotoxicity of [(125)I]ICF01035 and [(125)I]ICF01040 are not exclusively reliant on DNA alterations compatible with γ rays but likely result from local dose deposition (Auger electrons) leading to toxic compound leaks from acidic vesicles. In vivo, [(125)I]ICF01035 significantly reduced the number of B16F0 lung colonies, enabling a significant increase in survival of the treated mice. Targeting melanosomes or acidic vesicles is thus an option for future melanoma therapy. PMID:24691673

  8. 卟啉锰-TFO-吖啶的合成%Synthesis of manganese porphyrin-TFO-acridine

    Institute of Scientific and Technical Information of China (English)

    光丽霞; 袁发焕; 姜中兴; 王升启; 赵聪敏; 刘立; 温恩懿; 奚敏; 艾友萍; 管伟

    2003-01-01

    目的合成卟啉锰-TFO-吖啶化合物.方法合成含7-去氮-2-脱氧黄嘌呤的TFO;以吖啶修饰TFO的3′末端,以6碳氨基连接臂修饰TFO的5′末端;四苯基卟啉四羧酸经金属化、酯化后,与TFO的5′末端偶联;薄层层析纯化,质谱、紫外光光谱分析鉴定.结果薄层层析结果显示在对照寡核苷酸带前方有一条淡黄色的吖啶-TFO带;紫外光光谱分析显示卟啉锰-TFO-吖啶化合物同时具有260 nm处寡核苷酸的特征吸收峰和468 nm处卟啉锰的特征吸收峰.质谱分析结果证实,卟啉锰-TFO-吖啶化合物分子量实测值与理论值一致.结论合成了卟啉锰-TFO-吖啶化合物.

  9. Triple helix formation with purine-rich phosphorothioate-containing oligonucleotides covalently linked to an acridine derivative.

    OpenAIRE

    Lacoste, J; François, J C; Hélène, C

    1997-01-01

    Purine-rich (GA)- and (GT)-containing oligophosphorothioates were investigated for their triplex-forming potential on a 23 bp DNA duplex target. In our system, GA-containing oligophosphorothioates (23mer GA-PS) were capable of triplex formation with binding affinities lower than (GA)-containing oligophosphodiesters (23mer GA-PO). The orientation of the third strand 23mers GA-PS and GA-PO was antiparallel to the purine strand of the duplex DNA target. In contrast, (GT)-containing oligophosphor...

  10. 9,9-Dimethyl-12-(3-nitrophenyl-7,8,9,10,11,12-hexahydrobenz[a]acridin-11-one

    Directory of Open Access Journals (Sweden)

    Runhong Jia

    2009-09-01

    Full Text Available The title compound, C25H22N2O3, was synthesized by the reaction of 3-nitrobenzaldehyde, dimedone and 2-naphthylamine in ethanol. In the molecular structure, the cyclohexenone ring adopts an envelope conformation, whereas the piperidine ring has a boat conformation. The crystal packing is stabilized by intermolecular N—H...O hydrogen bonds.

  11. Triple helix formation: binding avidity of acridine-conjugated AG motif third strands containing natural, modified and surrogate bases opposed to pyrimidine interruptions in a polypurine target.

    OpenAIRE

    Orson, F M; Klysik, J; Bergstrom, D E; Ward, B; Glass, G A; P. Hua; Kinsey, B M

    1999-01-01

    A critical issue for the general application of triple-helix-forming oligonucleotides (TFOs) as modulators of gene expression is the dramatically reduced binding of short TFOs to targets that contain one or two pyrimidines within an otherwise homopurine sequence. Such targets are often found in gene regulatory regions, which represent desirable sites for triple helix formation. Using intercalator-conjugated AG motif TFOs, we compared the efficacy and base selectivity of 13 different bases or ...

  12. 9,9-Dimethyl-12-(3-nitro­phen­yl)-7,8,9,10,11,12-hexa­hydro­benz[a]acridin-11-one

    OpenAIRE

    Runhong Jia; Juhua Peng; Shujiang Tu

    2009-01-01

    The title compound, C25H22N2O3, was synthesized by the reaction of 3-nitrobenzaldehyde, dimedone and 2-naphthylamine in ethanol. In the molecular structure, the cyclohexenone ring adopts an envelope conformation, whereas the piperidine ring has a boat conformation. The crystal packing is stabilized by intermolecular N—H...O hydrogen bonds.

  13. FLUORIMETRIC DETERMINATION OF HYDROXYL FREE RADICAL PRODUCED BY FENTON REACTION WITH ACRIDINE RED%吖啶红荧光法检测Fenton反应产生的羟自由基

    Institute of Scientific and Technical Information of China (English)

    张爱梅; 臧运波

    2006-01-01

    提出检测Fenton反应产生羟自由基的方法.羟自由基与吖啶红发生反应后,吖啶红的荧光强度减弱,测定其荧光强度的变化,可间接测定羟自由基的产生量.试验结果表明,△F与吖啶红、硫酸铁及过氧化氢呈量效关系,发现抗氧化剂硫脲清除羟自由基作用呈明显的量效关系,方法稳定性好、操作简便、测定快速,测定了几种辛辣性食品的抗氧化性.

  14. Determination of lead ion by resonance energy transfer fluorescence quenching of acridine orange-rhodamine 6 G%吖啶橙-罗丹明6G共振能量转移荧光猝灭法测定铅

    Institute of Scientific and Technical Information of China (English)

    杨胜园; 徐小娜; 程健琳; 于军晖; 孙倩倩

    2015-01-01

    目的 根据吖啶橙和罗丹明6G两分子间能够有效地发生荧光共振能量转移,建立一种检测铅离子的新方法.方法 在表面活性剂SDS的存在下,吖啶橙和罗丹明6G分子间发生荧光共振能量转移,使罗丹明6G的荧光强度增大;加入铅离子后能对AO-R6 G体系的荧光产生猝灭作用,其荧光猝灭程度随着铅离子浓度的增大而增强,据此建立了检测Pb2+的新方法.结果 当pb2+浓度为2.0 ×10-7mol/L ~ 3.0×10-6mol/L时,与荧光猝灭程度△F有良好的线性关系,线性回归方程为y=16.34x +42.79,相关系数(r) =0.998,检出限为6.06×10-8mol/L,相对标准偏差(RSD)为3.4% ~5.3%,加标回收率为95.1% ~96.5%.结论 本方法操作简便、灵敏,可用于实际水样中铅含量的测定.

  15. 三维生物高分子氧化石墨烯复合凝胶对吖啶橙吸附性能的研究%STUDY ON ADSORPTION PROPERTY OF ACRIDINE ORANGE ON THE THREE-DIMENSIONAL BIOPOLYMER GRAPHENE OXIDE COMPOSITE GEL

    Institute of Scientific and Technical Information of China (English)

    王欢; 张萍; 王晓玲

    2016-01-01

    采用一种简单的共混冷冻法制备了三维生物高分子调控的氧化石墨烯复合水凝胶(3D/bio/GO/gel).通过傅里叶红外光谱、热重分析仪、粒度分析仪和荧光显微镜对氧化石墨烯复合水凝胶进行了表征.以吖啶橙(AO)为模型,研究了3D/bio/GO/gel对AO的吸附性能,并研究了羧甲基纤维钠的加入量和pH值对吸附性能的影响,实验结果表明,3D/bio/GO/gel对AO的吸附符合二级动力学模型和Freundlich模型.

  16. La coloración fluorescente con naranja de acridina y el PAP: validación de ambas técnicas para la detección de Trichomonas vaginalis FLUORESCENT STAINING WITH ACRIDINE ORANGE AND PAP SMEAR: VALIDATION TESTS OF BOTH TECNIQUES FOR THE DETECTION OF Trichomonas vaginalis

    OpenAIRE

    SIXTO RAUL COSTAMAGNA; MARIA PRADO FIGUEROA; OSCAR SORIA; ALEJANDRO FUENTES; RICARDO FERREYRA

    2000-01-01

    Se efectuó la validación de la coloración de Papanicolaou, utilizada para citología vaginal, frente a la coloración fluorescente con naranja de acridina, a fin de evaluar el valor de un resultado negativo para Trichomonas vaginalis obtenido en un PAP. Se estudiaron 80 muestras de flujo vaginal de mujeres entre 18 y 45 años, pacientes de consultorios externos de Ginecología del Hospital Municipal de la ciudad de Bahía Blanca, Provincia de Buenos Aires (Argentina). Las muestras se colorearon pa...

  17. 10-[2-(Dimethyl­amino)eth­yl]-9-(4-methoxy­phen­yl)-3,3,6,6-tetra­methyl-3,4,6,7,9,10-hexa­hydro­acridine-1,8(2H,5H)-dione

    OpenAIRE

    Balamurugan, P.; Jagan, R.; Thiagarajan, V.; M. Yamin, Bohari; Sivakumar, K.

    2009-01-01

    In the title compound, C28H38N2O3, the central ring of the acridinedione system adopts a boat conformation, while one of the outer rings adopts a half-chair conformation and the conformation of the other outer ring is between a sofa and a half-chair. The acridinedione system is buckled, with an angle of 22.01 (3)°. The crystal packing comprises layers of mol­ecules laid parallel to the ac plane, being reinforced by an intermolecular C—H⋯O interaction.

  18. In vitro mutagenicity testing. I. Kermide 601 resin, Sylgard 184 encapsulating resin, and Sylgard 184 curing agent. [Ames Salmonella assay system used

    Energy Technology Data Exchange (ETDEWEB)

    Wang, S.Y.; Smith, D.M.

    1978-08-01

    Five compounds, Kerimide 601 resin, Sylgard 184 encapsulating resin, Sylgard 184 curing agent, benzo(a)pyrene, and acridine orange were tested for in vitro mutagenicity using the Ames Salmonella assay system. Kerimide 601 resin, Sylgard 184 encapsulating resin, and Sylgard 184 curing agent were not mutagenic under the described experimental conditions, while benzo(a)pyrene and acridine orange were both mutagenic.

  19. Fate of Carbamazepine during Water Treatment

    DEFF Research Database (Denmark)

    Kosjek, T.; Andersen, Henrik Rasmus; Kompare, Boris;

    2009-01-01

    of acridone, hydroxy-(9H,10H)-acridine-9-carbaldehyde, acridone-N-carbaldehyde, and 1-(2-benzaldehyde)-(1H,3H-quinazoline-2,4-dione, while biological breakdown of acridine yielded acridone. In parallel, the transformation product iminostilbene was observed during sample analysis. In addition,this study...

  20. Bile canalicular cationic dye secretion as a model for P-glycoprotein mediated transport.

    Science.gov (United States)

    Thalhammer, T; Stapf, V; Gajdzik, L; Graf, J

    1994-04-01

    This study explores properties of P-glycoprotein dependent membrane transport in rat liver with the use of acridine orange as the substrate. We studied the biliary secretion of the dye, its binding to canalicular membrane P-glycoprotein, and effects of the inhibitor cyclosporin A: acridine orange is excreted into bile together with less hydrophobic and glucuronidated metabolites. Cyclosporin A inhibited both the secretion of acridine orange and of its metabolites. In TR- animals, a rat strain that is deficient of the canalicular multi-specific organic anion transport system, non-metabolized acridine orange is the predominant species in bile and its secretion is also inhibited by cyclosporin A. Binding of acridine orange to liver P-glycoprotein was analyzed by photoaffinity labeling with azidopine, a substrate of P-glycoprotein dependent transport in multi-drug resistant tumor cells. Labeling of the immunoprecipitated P-glycoprotein was inhibited by acridine orange, verapamil, and by cyclosporin A. The results show that biliary secretion of acridine orange is highly analogous to P-glycoprotein mediated membrane drug transport in tumor cells that exhibit multi-drug resistance.

  1. A spectroscopic study of interaction of cationic dyes with heparin

    Directory of Open Access Journals (Sweden)

    R. Nandini

    2010-01-01

    Full Text Available The interaction of two cationic dyes namely, acridine orange and pinacyanol chloride with an anionic polyelectrolyte, heparin, has been investigated by spectrophotometric method.The polymer induced metachromasy in the dyes resulting in the shift of the absorption maxima of the dyes towards shorter wavelengths. The stability of the complexes formed between acridine orange and heparin was found to be lesser than that formed between pinacyanol chloride and heparin. This fact was further confirmed by reversal studies using alcohols, urea and surfactants. The interaction of acridine orange with heparin has also been investigated fluorimetrically.The interaction parameters revealed that binding between acridine orange and heparin arises due to electrostatic interaction while that between pinacyanol chloride and heparin is found to involve both electrostatic and hydrophobic forces. The effect of the structure of the dye in inducing metachromasy has also been discussed.

  2. 9-(4-Bromophenoxycarbonyl-10-methylacridinium trifluoromethanesulfonate

    Directory of Open Access Journals (Sweden)

    Damian Trzybiński

    2010-06-01

    Full Text Available In the crystal structure of the title compound, C21H15BrNO2+·CF3SO3−, the cations form inversion dimers through π–π interactions between the acridine ring systems. These dimers are further linked by C—H...π and C—Br...π interactions. The cations and anions are connected by multidirectional C—H...O and C—F...π interactions. The acridine and benzene ring systems are oriented at 10.8 (1°. The carboxyl group is twisted at an angle of 85.2 (1° relative to the acridine skeleton. The mean planes of adjacent acridine units are parallel or almost parallel [inclined at an angle of 1.4 (1°] in the crystal structure.

  3. Evaluation of different methods for diagnosis of P. falciparum malaria

    Directory of Open Access Journals (Sweden)

    Mendiratta D

    2006-01-01

    Full Text Available Rapid diagnosis is a prerequisite for institution of effective treatment and reducing the mortality and morbidity of falciparum malaria. This study was taken up to compare the efficacy of various rapid methods viz, acridine orange, Plasmodium falciparum histidine rich protein II antigen detection and Field′s stain with traditional microscopy i.e., Leishman stain for diagnosing falciparum malaria. Thick and thin blood films of 443 consecutive patients with history of fever with chills and rigors were examined by Leishman and Field′s method. Acridine orange stained wet mounts of blood were examined under fluorescence microscopy. All films were examined by two independent microbiologists. Plasmodium falciparum histidine rich protein II antigen was detected using commercially available kit, Paracheck Pf. Out of the 443 subjects examined for P.falciparum 18.28% were detected by Leishman stain, 6.32% by Field′s stain, 18.28% by acridine orange and 18.1% by antigen based technique. Field′s stain missed 53 (65.4%, while Paracheck Pf was negative in 6(7.4% of the Leishman positive samples. All Field′s stain and acridine orange positives were positive by Leishman, but five Paracheck Pf positives were negative. Leishman stain is cost effective but if facilities are available one should use acridine orange for screening. The antigen detection kits are rapid, simple and are useful but to rule out false negatives in clinically suspected cases, Leishman stain is reliable.

  4. Synthesis, DNA Binding and Topoisomerase I Inhibition Activity of Thiazacridine and Imidazacridine Derivatives

    Directory of Open Access Journals (Sweden)

    Elizabeth Almeida Lafayette

    2013-12-01

    Full Text Available Thiazacridine and imidazacridine derivatives have shown promising results as tumors suppressors in some cancer cell lines. For a better understanding of the mechanism of action of these compounds, binding studies of 5-acridin-9-ylmethylidene-3-amino-2-thioxo-thiazolidin-4-one, 5-acridin-9-ylmethylidene-2-thioxo-thiazolidin-4-one, 5-acridin-9-ylmethylidene-2-thioxo-imidazolidin-4-one and 3-acridin-9-ylmethyl-thiazolidin-2,4-dione with calf thymus DNA (ctDNA by electronic absorption and fluorescence spectroscopy and circular dichroism spectroscopy were performed. The binding constants ranged from 1.46 × 104 to 6.01 × 104 M−1. UV-Vis, fluorescence and circular dichroism measurements indicated that the compounds interact effectively with ctDNA, both by intercalation or external binding. They demonstrated inhibitory activities to human topoisomerase I, except for 5-acridin-9-ylmethylidene-2-thioxo-1,3-thiazolidin-4-one. These results provide insight into the DNA binding mechanism of imidazacridines and thiazacridines.

  5. Organic dyes removal using magnetically modified rye straw

    Energy Technology Data Exchange (ETDEWEB)

    Baldikova, Eva, E-mail: baldie@email.cz [Department of Nanobiotechnology, Institute of Nanobiology and Structural Biology of GCRC, Na Sadkach 7, 370 05 Ceske Budejovice (Czech Republic); Safarikova, Mirka [Department of Nanobiotechnology, Institute of Nanobiology and Structural Biology of GCRC, Na Sadkach 7, 370 05 Ceske Budejovice (Czech Republic); Safarik, Ivo, E-mail: ivosaf@yahoo.com [Department of Nanobiotechnology, Institute of Nanobiology and Structural Biology of GCRC, Na Sadkach 7, 370 05 Ceske Budejovice (Czech Republic); Regional Centre of Advanced Technologies and Materials, Palacky University, Slechtitelu 11, 783 71 Olomouc (Czech Republic)

    2015-04-15

    Rye straw, a very low-cost material, was employed as a biosorbent for two organic water-soluble dyes belonging to different dye classes, namely acridine orange (acridine group) and methyl green (triarylmethane group). The adsorption properties were tested for native and citric acid–NaOH modified rye straw, both in nonmagnetic and magnetic versions. The adsorption equilibrium was reached in 2 h and the adsorption isotherms data were analyzed using the Langmuir model. The highest values of maximum adsorption capacities were 208.3 mg/g for acridine orange and 384.6 mg/g for methyl green. - Highlights: • Rye derivatives can be considered as efficient adsorbents for organic dyes. • Magnetic modification of straw by microwave-synthesized magnetic iron oxides. • Citric acid–NaOH modification increased the maximum adsorption capacities.

  6. 9-Benzyl-10-methylacridinium trifluoromethanesulfonate

    Directory of Open Access Journals (Sweden)

    Damian Trzybiński

    2010-07-01

    Full Text Available In the crystal structure of the title compound, C21H18N+·CF3OS3−, the cations form inversion dimers through π–π interactions between the acridine ring systems. These dimers are further linked by C—H...π interactions. The cations and anions are connected by C—H...O, C—F...π and S—O...π interactions. The acridine and benzene ring systems are oriented at a dihedral angle of 76.8 (1°with respect to each other. The acridine moieties are either parallel or inclined at an angle of 62.4 (1° in the crystal structure.

  7. 9-(4-Chlorophenoxycarbonyl-10-methylacridinium trifluoromethanesulfonate

    Directory of Open Access Journals (Sweden)

    Damian Trzybiński

    2010-11-01

    Full Text Available In the crystal of the title compound, C21H15ClNO2+·CF3SO3−, adjacent cations are linked through C—H...π and π–π interactions [centroid–centroid distance = 3.987 (2 Å], and neighboring cations and anions via C—H...O and C—F...π interactions. The acridine ring system and benzene ring are oriented at a dihedral angle of 1.0 (1° while the carboxyl group is twisted at an angle of 85.0 (1° relative to the acridine skeleton. The mean planes of adjacent acridine units are either parallel or inclined at an angle of 78.2 (1° in the crystal structure.

  8. 10-Methyl-9-[2-(propan-2-ylphenoxycarbonyl]acridinium trifluoromethanesulfonate

    Directory of Open Access Journals (Sweden)

    Damian Trzybiński

    2010-11-01

    Full Text Available In the crystal of the title compound, C24H22NO2+·CF3SO3−, adjacent cations and anions are connected through C—H...O, C—H...F and S–O...π interactions, while neighboring cations via π–π interactions [centroid–centroid distance = 3.962 (2 Å]. The acridine and benzene ring systems are oriented at a dihedral angle of 14.6 (1°. The carboxyl group is twisted at an angle of 87.6 (1° relative to the acridine skeleton. The mean planes of adjacent acridine units are parallel or inclined at an angle of 13.4 (1° in the crystal structure.

  9. Complexes of nitracrine with DNA. Stoichiometry of binding.

    Science.gov (United States)

    Szmigiero, L; Gniazdowski, M

    1981-01-01

    In the presence of sulfhydryl compounds, an anticancer drug 1-nitro-9-(3-N,N-dimethylaminopropylamino) acridine (nitracrine, Ledakrin) forms irreversible complexes of decreased template activity with DNA. Stoichiometry of the complexes was estimated using the drug labelled with 14C in the acridine ring or in the propyl chain and with 3H in the acridine ring. Up to 50 irreversibly bound 14C-nitracrine molecules were found per 10(3) nucleotides of calf thymus DNA in the presence of dithiothreitol (DTT). Considerably lower binding observed using 3H-labelled drug, particularly when the complexes were formed in the presence of mercaptoethanol (ME) indicates that substitution of tritium atoms occurred during the reaction. Relationship between stoichiometry and template activity in RNA synthesis in vitro system of the complexes was estimated in the paper. PMID:7198467

  10. Effects and uptake of polycyclic aromatic compounds in snails (Helix aspersa).

    Science.gov (United States)

    Sverdrup, Line Emilie; De Vaufleury, Annette; Hartnik, Thomas; Hagen, Snorre B; Loibner, Andreas Paul; Jensen, John

    2006-07-01

    The International Standardization Organization recently launched a soil toxicity test with snails (Helix aspersa). We assessed the sensitivity of this test for seven polycyclic aromatic compounds. Control animals had 100% survival and low variability for growth measurements. Maximum exposure concentrations of 2800 mg/kg (4000 mg/kg for acridine) had no effect on survival. Similarly, growth (biomass and shell size) was not affected by pyrene, fluoranthene, fluorene, carbazole, phenanthrene, or acridine, whereas dibenzothiophene gave a 10% effect concentration of 1600 mg/kg. Measured internal concentrations of carbazole, dibenzothiophene, and acridine increased with increasing soil concentrations, but biota-soil accumulation factors were low (0.002-0.1). Compared to previously tested organisms, with all being exposed in the same soil type and under similar test conditions, the H. aspersa test was relatively insensitive to all substances.

  11. Organic dyes removal using magnetically modified rye straw

    International Nuclear Information System (INIS)

    Rye straw, a very low-cost material, was employed as a biosorbent for two organic water-soluble dyes belonging to different dye classes, namely acridine orange (acridine group) and methyl green (triarylmethane group). The adsorption properties were tested for native and citric acid–NaOH modified rye straw, both in nonmagnetic and magnetic versions. The adsorption equilibrium was reached in 2 h and the adsorption isotherms data were analyzed using the Langmuir model. The highest values of maximum adsorption capacities were 208.3 mg/g for acridine orange and 384.6 mg/g for methyl green. - Highlights: • Rye derivatives can be considered as efficient adsorbents for organic dyes. • Magnetic modification of straw by microwave-synthesized magnetic iron oxides. • Citric acid–NaOH modification increased the maximum adsorption capacities

  12. Inhibition of DNA topoisomerase I activity and induction of apoptosis by thiazacridine derivatives

    Energy Technology Data Exchange (ETDEWEB)

    Barros, Francisco W.A. [Department of Physiology and Pharmacology, School of Medicine, Federal University of Ceará, Fortaleza, Ceará (Brazil); Bezerra, Daniel P., E-mail: danielpbezerra@gmail.com [Department of Physiology, Federal University of Sergipe, São Cristóvão, Sergipe (Brazil); Ferreira, Paulo M.P. [Department of Biological Sciences, Federal University of Piauí, Picos, Piauí (Brazil); Cavalcanti, Bruno C. [Department of Physiology and Pharmacology, School of Medicine, Federal University of Ceará, Fortaleza, Ceará (Brazil); Silva, Teresinha G.; Pitta, Marina G.R.; Lima, Maria do C.A. de; Galdino, Suely L.; Pitta, Ivan da R. [Department of Antibiotics, Federal, University of Pernambuco, Recife, Pernembuco (Brazil); Costa-Lotufo, Letícia V.; Moraes, Manoel O. [Department of Physiology and Pharmacology, School of Medicine, Federal University of Ceará, Fortaleza, Ceará (Brazil); Burbano, Rommel R. [Institute of Biological Sciences, Federal University of Pará, Belém, Pará (Brazil); Guecheva, Temenouga N.; Henriques, João A.P. [Biotechnology Center, Federal University of Rio Grande do Sul, Porto Alegre, Rio Grande do Sul (Brazil); Pessoa, Cláudia, E-mail: cpessoa@ufc.br [Department of Physiology and Pharmacology, School of Medicine, Federal University of Ceará, Fortaleza, Ceará (Brazil)

    2013-04-01

    Thiazacridine derivatives (ATZD) are a novel class of cytotoxic agents that combine an acridine and thiazolidine nucleus. In this study, the cytotoxic action of four ATZD were tested in human colon carcinoma HCT-8 cells: (5Z)-5-acridin-9-ylmethylene-3-(4-methylbenzyl)-thiazolidine-2,4-dione — AC-4; (5ZE)-5-acridin-9-ylmethylene-3-(4-bromo-benzyl)-thiazolidine-2,4-dione — AC-7; (5Z)-5-(acridin-9-ylmethylene)-3-(4-chloro-benzyl) -1,3-thiazolidine-2,4-dione — AC-10; and (5ZE)-5-(acridin-9-ylmethylene)-3-(4-fluoro-benzyl)-1,3-thiazolidine-2, 4-dione — AC-23. All of the ATZD tested reduced the proliferation of HCT-8 cells in a concentration- and time-dependent manner. There were significant increases in internucleosomal DNA fragmentation without affecting membrane integrity. For morphological analyses, hematoxylin–eosin and acridine orange/ethidium bromide were used to stain HCT-8 cells treated with ATZD, which presented the typical hallmarks of apoptosis. ATZD also induced mitochondrial depolarisation and phosphatidylserine exposure and increased the activation of caspases 3/7 in HCT-8 cells, suggesting that this apoptotic cell death was caspase-dependent. In an assay using Saccharomyces cerevisiae mutants with defects in DNA topoisomerases 1 and 3, the ATZD showed enhanced activity, suggesting an interaction between ATZD and DNA topoisomerase enzyme activity. In addition, ATZD inhibited DNA topoisomerase I action in a cell-free system. Interestingly, these ATZD did not cause genotoxicity or inhibit the telomerase activity in human lymphocyte cultures at the experimental levels tested. In conclusion, the ATZD inhibited the DNA topoisomerase I activity and induced tumour cell death through apoptotic pathways. - Highlights: ► Thiazacridine derivatives induce mitochondrial-dependent apoptotic cell death. ► Thiazacridine derivatives inhibit DNA topoisomerase I action. ► Thiazacridine derivatives failed to cause genotoxicity on human lymphocytes.

  13. Design and synthesis of threading intercalators to target DNA.

    Science.gov (United States)

    Howell, Lesley A; Gulam, Rosul; Mueller, Anja; O'Connell, Maria A; Searcey, Mark

    2010-12-01

    Threading intercalators are high affinity DNA binding agents that bind by inserting a chromophore into the duplex and locating one group in each groove. The first threading intercalators that can be conjugated to acids, sulfonic acids and peptides to target them to duplex DNA are described, based upon the well studied acridine-3- or 4-carboxamides. Cellular uptake of the parent acridine is rapid and it can be visualized in the nucleus of cells. Both the parent compounds and their conjugates maintain antitumor activity.

  14. Light-controlled mass formation of aggregates of molecules in organic compounds

    Institute of Scientific and Technical Information of China (English)

    Tariel D.Ebralidze; Nadia A.Ebralidze; Giorgi A.Mumladze; Enriko S.Kitsmarishvili

    2009-01-01

    During the mass formation of aggregates of molecules in a gelatin film dyed with the mixture of chrysophenine and acridine yellow dyes,photo-reorientation,photo-disorientation,and photo-orientation of the molecules are observed.Based on these observations,the photo-induction of granular aniso tropy may be realized.

  15. Comparison of four diagnostic techniques for detection of Trichomonas vaginalis infection in females attending tertiary care hospital of North India

    Directory of Open Access Journals (Sweden)

    Razia Khatoon

    2015-01-01

    Full Text Available Background: Trichomonas vaginalis causes a common sexually transmitted disease trichomoniasis, which may lead to increased risk of transmission of human immunodeficiency virus infection and other pelvic inflammatory diseases. Wet mount examination is the most common test for diagnosis, but it has low sensitivity. Acridine orange staining can be used for diagnosis, but it requires special microscopic facility. Culture is considered as the gold standard, but it takes a long time for diagnosis. OSOM Trichomonas Rapid Test is a recently introduced rapid method based on immunochromatographic assay of trichomonal protein antigens. Hence, the present study was done to compare these four diagnostic techniques for detection of trichomoniasis in females with vaginal discharge. Materials and Methods: Vaginal swabs were taken from 835 female patients and wet mount examination, acridine orange staining, culture in Kupferberg medium, and OSOM Trichomonas Rapid Test, were performed. Results: Out of 835 patients included in our study, 68 (8.1% positive cases of trichomoniasis were detected by culture. OSOM Trichomonas Rapid Test detected 63 (7.5% cases, acridine orange staining detected 53 (6.3% cases, whereas, wet mount examination detected only 45 (5.4% positive cases. OSOM Trichomonas Rapid Test performed well and showed high sensitivity and specificity of 88.2% and 99.6%, respectively. Conclusion: As OSOM Trichomonas Rapid Test is a point of care test and gave better results than both wet mount examination and acridine orange staining; it can be used as a routine test in peripheral areas lacking laboratory facilities.

  16. A novel immunoradiometric assay for human liver ferritin.

    OpenAIRE

    Al-Shawi, A; Dawnay, A; Landon, J

    1983-01-01

    Rivanol, the cationic salt of an acridine base, has been used as a novel separation procedure in an immunoradiometric assay for human liver ferritin. The separation step is based on the differences in charge and molecular weight between the labelled antibody-ferritin complex and free labelled immunoglobulins. The resultant assay is simple, reproducible and sufficiently sensitive to determine serum concentrations of ferritin.

  17. Determination of Dihydrobenzoacridinone Structures by NMR, IR, and UV Spectroscopy and Mass Spectrometry

    Science.gov (United States)

    Kozlov, N. G.; Zhiharko, Yu. D.; Skakovsky, E. D.; Baranovsky, A. V.; Ogorodnikova, M. M.; Basalaeva, L. I.

    2016-01-01

    Condensation of 2-naphthylamine, aromatic aldehydes, and dimedone was found to produce 9,10-dihydrobenzo[a] acridin-11-one derivatives according to PMR, 13C NMR, and IR spectroscopy and mass spectrometry. Correlation spectroscopy showed that the carbonyl in the synthesized dihydrobenzoacridinone derivatives was located on C11.

  18. Combination and cleavage of HBV DNA fragments by triple helix-forming oligonucleotides modified with manganese porphyrin in vitro

    Institute of Scientific and Technical Information of China (English)

    光丽霞; 袁发焕; 奚敏; 赵聪敏; 刘立; 温恩懿; 艾友萍

    2003-01-01

    Objective To observe the ability of triple helix-forming oligonucleotides (TFOs) modified with manganese porphyrin to combine with and cleave HBV DNA fractions. Methods TFO were modified with manganese porphyrin and acridines, and then reacted with the 32P labeled HBV DNA fragments at 37℃ in vitro (pH 7.4). Electrophoretic mobility shift assays and Dnase Ⅰ footprinting tests were used to show the affinity and specificity of TFO to bind to target sequences. The ability of TFO to cleave HBV DNA fragments was tested by cleavage experiments. Results TFO modified with manganese porphyrin and acridine could bind to the target sequence in a sequence-dependent manner, with a Kd value of 3.5×10-7 mol/L and a relative affinity of 0.008. In the presence of potassium monopersulfate (KHSO5), TFO modified with manganese porphyrin and acridine could cleave the target sequence where the triplex DNA was formed. Conclusion In the presence of KHSO5, TFO modified with manganese porphyrin and acridine could bind and cleave the target HBV-DNA in a sequence-dependent manner.

  19. Synthesis of Thioacridine Derivatives Using Lawesson’s Reagent

    OpenAIRE

    Palla Mahesh; B.Dilip Kumar; B. Rama Devi; Murthy, Y. L. N.

    2015-01-01

    The synthesis of thioacridine derivatives (5a-j) have been achieved by the reaction of acridines (4a-j) with Lawesson’s reagent in toluene under refluxing conditions to yield products in high yields. The yields of the products are promising and the products are characterized by advanced spectroscopic studies.

  20. One-pot synthesis of novel 1, 8-dioxo-decahydroacridines containing phenol and benzamide moiety and their synthetic uses

    Indian Academy of Sciences (India)

    Ali Dorehgiraee; Esmat Tavakolinejad Kermani; Hojatollah Khabazzadeh

    2014-07-01

    An efficient synthesis of some new 1, 8-dioxo-decahydroacridines is achieved via one-pot, threecomponent condensation of aromatic aldehydes, cyclic diketone, and 4-amino benzamide/4-aminophenol. Reaction of these acridines with dimethylacetylene dicarboxylate and triphenylphosphine or cyclohexylisocyanide gives stable phosphorus ylides or 4H-chromene derivatives, respectively, with good yields.

  1. Microscale response of sediment variables to benthic disturbance in the Central Indian Ocean Basin

    Digital Repository Service at National Institute of Oceanography (India)

    Nair, S.; Mohandass, C.; LokaBharathi, P.A.; Sheelu, G.; Raghukumar, C.

    was filtered on a 0.2-m (pore- size) Nucleopore polycarbonate membrane to which 0.1% acridine orange was added (Hob-bie et al.,1977); after the membrane was washed,the bacteria were counted with anepifluorescence microscope. The bacterial numbers are expresed...

  2. Dormant barley aleurone shows heterogeneity and a specific cytodifferentiation

    NARCIS (Netherlands)

    Schuurink, R.C.; Bakhuizen, R.; Libbenga, K.R.; Boulanger, F.; Sinjorgo, K.M.C.

    1997-01-01

    In response to gibberellic acid, aleurone layers isolated from dormant barley (Hordeum distichum L. cv. Triumph) kernels produced significantly less alpha-amylase than aleurones from non-dormant kernels. Light microscopical investigations using the dye acridine orange as well as electron microscopic

  3. 9-Phenyl-10H-acridinium trifluoromethanesulfonate

    Directory of Open Access Journals (Sweden)

    Damian Trzybiński

    2010-11-01

    Full Text Available In the crystal structure of the title compound, C19H14N+·CF3SO3−, the cations are linked to each other by very weak C—H...π interactions, while the cations and anions are connected by N—H...O, C—H...O and S—O...π interactions. The acridine ring system and the phenyl ring are oriented at an angle of 80.1 (1° with respect to each other. The mean planes of adjacent acridine units are either parallel or inclined at an angle of 35.6 (1°. The trifluoromethanesulfonate anions are disordered over two positions; the site occupancy factors are 0.591 (8 and 0.409 (8.

  4. 2-Methoxy-9-phenoxyacridine

    Directory of Open Access Journals (Sweden)

    Damian Trzybiński

    2010-04-01

    Full Text Available The molecules in the crystal structure of the title compound, C20H15NO2, form inversion dimers connected through the C—H...N and π–π interactions. These dimers are further linked by C—H...π interactions. The methoxy group is nearly coplanar with the acridine ring system [dihedral angle = 4.5 (1°], whereas the phenoxy fragment is nearly perpendicular to it [dihedral angle = 85.0 (1°]. The mean planes of the acridine ring systems are either parallel or inclined at angles of 14.3 (1, 65.4 (1 and 67.3 (1° in the crystal.

  5. Survival and activity of Streptococcus faecalis and Escherichia coli in tropical freshwater

    Energy Technology Data Exchange (ETDEWEB)

    Muniz, I.; Jimenez, L.; Toranzos, G.A.; Hazen, T.C. [Univ. of Puerto Rico, Rio Piedras (Puerto Rico)

    1988-12-31

    The survival of Streptococcus facecalis and Escherichia coli was studied in situ in a tropical rain forest watershed using membrane diffusion chambers. Densities were determined by acridine orange direct count and Coulter Counter. Population activity was determined by microautoradiography, cell respiration, and by nucleic acid composition. Densities of S. facecalis and E. coli decreased less than 1 log unit after 105 h as measured by direct count methods. Activity as measured by respiration, acridine orange activity, and microautoradiography indicated that both bacteria remained moderately active during the entire study. After 12 h, E. coli was more active than S. faecalis as measured by nucleic acid composition. E. coli and S. faecalis survived and remained active for more than 5 days. Consequently, both would seem to be unsuitable as indicators of recent fecal contamination in tropical waters.

  6. Quadruplex-targeting anticancer drug BRACO-19 voltammetric and AFM characterization

    International Nuclear Information System (INIS)

    The quadruplex-targeting anticancer drug BRACO-19 adsorption and redox behaviour were investigated by atomic force microscopy (AFM) on a highly oriented pyrolytic graphite surface and by cyclic, differential pulse and square-wave voltammetry at a glassy carbon electrode. The AFM and voltammetric results demonstrated that the BRACO-19 orientation and strong adsorption, with the acridine aromatic core parallel or perpendicular to the carbon electrode surface depending on solution pH, directly influences the peak potentials and redox behaviour. BRACO-19 oxidation was a complex, pH-dependent, four-step electrode process. The first oxidation step was reversible, the second, third and fourth oxidation steps irreversible, and an electroactive irreversibly oxidized BRACO-19 oxidation product was formed. BRACO-19 reduction occurred in two irreversible, pH-independent steps. The proposed redox mechanisms are related to the pyrrolidine and acridine moieties

  7. Effect of DNA-interacting drugs on phage T7 RNA polymerase.

    Science.gov (United States)

    Piestrzeniewicz, M; Studzian, K; Wilmańska, D; Płucienniczak, G; Gniazdowski, M

    1998-01-01

    9-Aminoacridine carboxamide derivatives studied here form with DNA intercalative complexes which differ in the kinetics of dissociation. Inhibition of total RNA synthesis catalyzed by phage T7 and Escherichia coli DNA-dependent RNA polymerases correlates with the formation of slowly dissociating acridine-DNA complex of time constant of 0.4-2.3 s. Their effect on RNA synthesis is compared with other ligands which form with DNA stable complexes of different steric properties. T7 RNA polymerase is more sensitive to distamycin A and netropsin than the E. coli enzyme while less sensitive to actinomycin D. Actinomycin induces terminations in the transcript synthesized by T7 RNA polymerase. Despite low dissociation rates of DNA complexes with acridines and pyrrole antibiotics no drug dependent terminations are observed with these ligands. PMID:9701505

  8. Kinetics and mechanism of desulfurization and denitrogenation of coal-derived liquids. Ninth quarterly report, June 21, 1977--September 20, 1977

    Energy Technology Data Exchange (ETDEWEB)

    Gates, B. C.; Katzer, J. R.; Olson, J. H.; Kwart, H.; Stiles, A. B.

    1978-05-01

    Flow reactor studies of HDS of dibenzothiophene at about 300/sup 0/C and 100 atm have shown a new dependence of catalyst activity on H/sub 2/S concentration at low values. We infer that the catalyst undergoes structural changes during reaction, sometimes becoming sulfur deficient and relatively inactive. It is evidently necessary to maintain the catalyst in a properly sulfided state during reactor startup and operation by avoiding its contacting too little H/sub 2/S. The relative activities of Ni--Mo/Al/sub 2/O/sub 3/, CO--Mo/Al/sub 2/O/sub 3/, and Ni--W/Al/sub 2/O/sub 3/ catalysts for HDN of acridine have been determined, showing that Ni--Mo/Al/sub 2/O/sub 3/ is the most active. The results are interpreted inthe context of the acridine reaction network, in which both the hydrogenation and hydrogenolysis (cracking) reactions are kinetically important.

  9. Viability of bacteria in dental calculus - A microbiological study

    Directory of Open Access Journals (Sweden)

    Moolya Nikesh

    2010-01-01

    Full Text Available Aim: The aim of this study was (1 To investigate the viability of bacteria within supragingival and subgingival calculus, (2 To examine motility of bacteria, and (3 To identify bacterial morphotypes in calculus. Materials and Methods: Supra and subgingival calculus were harvested from 30 subjects having clinical evidence of chronic inflammatory periodontal disease and were divided into two groups . Samples from both groups were immediately transported to the Department of Microbiology for gram staining, acridine orange staining, bacterial culture and to the Department of Oral Pathology for dark field microscopy. Results: Gram staining revealed presence of bacteria within the samples. Dark field microscopic examination revealed presence of filamentous organisms, spirochetes, and motile short bacilli. Acridine orange fluorescent stain showed that the viable bacteria appeared apple green. Bacterial culture revealed presence of a variety of aerobic organisms. Conclusion: From the results, it appeared that viable bacteria were present within calculus especially within internal channels and lacunae.

  10. Endovascular photodynamic therapy to prevent arterial restenosis

    OpenAIRE

    Gabeler, E.E.E.

    2003-01-01

    markdownabstract__Abstract__ Since their existence, man has appreciated the benefits of sunlight and described some of its medicinal effects known as heliotherapy. Herodotus in the 6th century BC noticed that sunlight had beneficial effects on bone growth. Hippocrates in 460-375 BC advocated the use of heliotherapy for various human maladies [1]. In 1898, McCall-Anderson described skin photosensitivity due to porphyrin molecules [2]. In 1900, Raab using acridine orange described a photochemic...

  11. DNA-Conjugated Organic Chromophores in DNA Stacking Interactions

    DEFF Research Database (Denmark)

    Filichev, Vyacheslav V.; Pedersen, Erik Bjerregaard

    2009-01-01

    Since the discovery of the intercalation of acridine derivatives into DNA (1961), chemists have synthesized many intercalators tethered to DNA. Advances in the chemical synthesis of modified nucleosides along with progress in oligonucleotide synthesis have made it possible to introduce organic...... review presents those efforts in the design of intercalators/organic chromophores as oligonucleotide conjugates that form a foundation for the generation of novel nucleic acid architectures...

  12. Transmissible Resistance to Penicillin G, Neomycin, and Chloramphenicol in Rhizobium japonicum1

    Science.gov (United States)

    Cole, Michael A.; Elkan, Gerald H.

    1973-01-01

    The genetic basis for resistance to a number of antibiotics was examined in Rhizobium japonicum. Resistance to penicillin G, neomycin, and chloramphenicol appears to be mediated by an extrachromosomal element similar to that found in the Enterobacteriaceae. Resistance to these antibiotics was eliminated from cells by treatment with acridine orange, and resistance to all three antibiotics could be transferred en bloc to Agrobacterium tumefaciens under conditions excluding transformation or transduction as possible genetic mechanisms. PMID:4491197

  13. Hepatitis in skunks caused by the virus of infectious canine hepatitis.

    Science.gov (United States)

    Karstad, L; Ramsden, R; Berry, T J; Binn, L N

    1975-10-01

    Two cases of acute, fatal, hepatitis occurred in young, striped skunks (Mephitis mephitis) trapped in southern Ontario. Histologically, lesions in the liver were similar to infectious canine hepatitis. A virus was isolated which produced large intranuclear inclusions in dog kidney cell cultures. These inclusions were Feulgen-positive and fluoresced green with acridine orange stain. The skunk hepatitis isolate was identified as the virus of infectious canine hepatitis by virus neutralization tests. PMID:172663

  14. Enumeration and Biomass Estimation of Bacteria in Aquifer Microcosm Studies by Flow Cytometry

    OpenAIRE

    DeLeo, P. C.; Baveye, P

    1996-01-01

    Flow cytometry was used to enumerate and characterize bacteria from a sand column microcosm simulating aquifer conditions. Pure cultures of a species of Bacillus isolated from subsurface sediments or Bacillus megaterium were first evaluated to identify these organisms' characteristic histograms. Counting was then carried out with samples from the aquifer microcosms. Enumeration by flow cytometry was compared with more-traditional acridine orange direct counting. These two techniques gave stat...

  15. Identification keys on rattans (Calamus spp.) from Central Sulawesi based on anatomical structure of stems

    OpenAIRE

    ANDI TANRA TELLU

    2005-01-01

    This study was conducted to obtain information the anatomical characteristics of 20 rattan species from Central Sulawesi and to use it for anatomical identification of rattan species. The rattan comprised 16 Calamus species, three Daemonorops species and one Korthalsia species. For anatomical observation 10-15 mm pieces of the mature stem from shares of tip do not have frond were processed with polyethilene glycol 2000, cut at 18-32 µm and stained with a combination of acridin-cryzoidin red a...

  16. Molecular modeling study of intercalation complexes of tricyclic carboxamides with d(CCGGCGCCGG) and d(CGCGAATTCGCG)

    OpenAIRE

    Varvaresou, Athanasia; Iakovou, Kriton

    2010-01-01

    Abstract Tricyclic dyes with different mesoatoms such as xanthenes (fluorescein, eosin) anthracenes and acridines (proflavine) approved by the Food and Drug Administration (FDA) for use in foods, pharmaceuticals and cosmetic preparations interact with DNA, and some of them do so through intercalation. Hyperchem 7.5, Spartan 04, Yasara 10.5.14 program packages and molecular modeling, molecular mechanics and dynamics techniques with the oligonucleotides d(CCGGCGCCGG)2 and d(CGCGAATTC...

  17. A comparison of herpes simplex virus plaque development after viral treatment with anti-DNA or antilipid agents.

    OpenAIRE

    Coohill, T P; Babich, M; Taylor, W.D.; Snipes, W

    1980-01-01

    The plaque development of Herpes simplex virus type 1 (HSV) is slower for viruses treated with two anti-DNA agents: ultraviolet radiation (UV) or n-acetoxy-2-acetyl-aminofluorene. For HSV treated with three antimembrane agents--butylated hydroxytoluene, acridine plus near UV radiation, or ether--the plaque development time is the same as for untreated viruses. These differences hold even for viruses that survived treatment that lowered viability below the 1% level. Gamma ray inactivation of H...

  18. Comparison of different diagnostic techniques in Plasmodium falciparum cerebral malaria

    OpenAIRE

    Fatima Shujatullah, Abida Malik, Haris M. Khan & Ashraf Malik

    2006-01-01

    Background & objectives: Plasmodium falciparum cerebral malaria remains a major health problemin India. The efficacy of treatment of cerebral malaria lies in its early diagnosis through rapid diagnosticmethods. ParaSights-F test detects HRP-2 antigen secreted by parasitised red blood cells andquantitative buffy coat assay (QBC) is examination of buffy coat for the presence of malarial parasitestained with acridine orange. This study was performed to evaluate the effectiveness of ParaSight-F t...

  19. Anionic Lipids Enriched at the ExPortal of Streptococcus pyogenes▿

    OpenAIRE

    Rosch, Jason W.; Hsu, Fong Fu; Caparon, Michael G.

    2006-01-01

    The ExPortal of Streptococcus pyogenes is a membrane microdomain dedicated to the secretion and folding of proteins. We investigated the lipid composition of the ExPortal by examining the distribution of anionic membrane phospholipids. Staining with 10-N-nonyl-acridine orange revealed a single microdomain enriched with an anionic phospholipid whose staining characteristics and behavior in a cardiolipin-deficient mutant were characteristic of phosphatidylglycerol. Furthermore, the location of ...

  20. Quenching of fluorescence by crystal violet and its use to differentiate between surface-bound and internalized bacteria

    Science.gov (United States)

    Mathew, S.; Lim, Y. C.; Kishen, A.

    2008-06-01

    Phagocytosis is a complex process involving attachment, ingestion and intracellular processing of bacteria by phagocytes. A great difficulty in the evaluation of this process is to differentiate between attachment of the particles to the cell surface and internalization of the particles by the cells. Various techniques have been used to differentiate internalized and surface-attached bacteria in cultured cells, but only a few permit differentiations between surface-bound and internalized bacteria. In this study the quenching of fluorescence by crystal violet on acridine orange stained bacterial biofilm and planktonic bacterial cells is used to differentiate between surface-bound and internalized bacteria within macrophages. Method: One week old Enterococcus faecalis biofilm was grown on perspex and glass substrates in All-Culture medium (nutrient-rich condition) and phosphate buffered saline (nutrient-deprived condition). As model systems, human monocytic (THP-1) and histiocytic (U937) cell lines were used. These cell lines were incubated with the biofilm bacteria for 4 hrs in CO II incubator at 37 °C. The cells and bacteria were stained with acridine orange and quenched with crystal violet to distinguish between surface-bound and internalized bacteria. Results: The presence of green-fluorescing internalized bacteria was detected within the macrophages under the planktonic, nutrient-rich and nutrient-deprived biofilm conditions. All infecting bacteria take up acridine orange and fluoresced green, crystal violet quenched the fluorescence of extra-cellular adhering bacteria so that only fluorescent intracellular bacteria would be visible under fluorescent light microscopy.

  1. Antiparasitic activities of acridone alkaloids from Swinglea glutinosa (Bl.) Merr

    International Nuclear Information System (INIS)

    Eleven acridone alkaloids isolated from Swinglea glutinosa (Bl.) Merr. were examined for in vitro activity against chloroquine-sensitive Plasmodium falciparum 3D7, Trypanosoma brucei rhodesiense STIB900 and Leishmania donovani L82. An assay with KB cells was developed in order to compare in vitro toxicity of alkaloids with the selective action on the parasites. Nine of the compounds had IC50 values ranging from 0.3 to 11.6 μM against P. falciparum. In contrast, a small number of compounds showed significant activity against T. brucei rhodesiense and none had activity against L. donovani. Among the alkaloids three had IC50 50 < 10 μM. The characterization of the acridone alkaloids, 1,3,5-trihydroxy-4-methoxy-10-methyl-2,8-bis(3-methylbut-2-enyl)acridin-9 (10H)-one (1), 2,3-dihydro-4,9-dihydroxy-2-(2-hydroxypropan-2-yl)-11-methoxy-10-methylfuro [3,2-b] acridin-5(10H)-one (2) and 3,4-dihydro-3,5,8-trihydroxy-6-methoxy-2,2,7-trimethyl-2Hpyrano[ 2,3-a]acridin-12(7H)-one (3), is discussed, as well as the structure-activity relationship of all compounds assayed. Isolation and spectral data of alkaloids 1-3 are described for the first time although their cytotoxicities to cancer cells have been described before. (author)

  2. In vitro cytotoxicity activity on several cancer cell lines of acridone alkaloids and N-phenylethyl-benzamide derivatives from Swinglea glutinosa (Bl.) Merr.

    Science.gov (United States)

    Braga, P A C; Dos Santos, D A P; Da Silva, M F D G F; Vieira, P C; Fernandes, J B; Houghton, P J; Fang, R

    2007-01-01

    The methanol extract from the stems and fruits of Swinglea glutinosa (Rutaceae) afforded 11 known acridone alkaloids and three N-phenylethyl-benzamide derivatives, glycocitrine-IV, 1,3,5-trihydroxy-4-methoxy-10-methyl-2,8-bis(3-methylbut-2-enyl)acridin-9(10H)-one, 1,3,5- trihydroxy-2,8-bis(3-methylbut-2-enyl)-10-methyl-9-acridone, citbrasine, citrusinine-II, citrusinine-I, 5-dihydroxyacronycine, pyranofoline, 3,4-dihydro-3,5,8-trihydroxy-6-methoxy-2,2,7-trimethyl-2H-pyrano[2,3-a]acridin-12(7H)-one, 2,3-dihydro-4,9-dihydroxy-2-(2-hydroxy-propan-2-yl)-11-methoxy-10-methylfuro[3,2-b]acridin-5(10H)-one, bis-5-hydroxyacronycine, N-(2-{4-[(3,7-dimethylocta-2,6-dien-1-yl)oxy]phenyl}ethyl)benzamide, N-(2-{4-[(3,7-dimethyl-4-acethyl-octa-2,6-dien-1-yl)oxy]phenyl}ethyl)benzamide, and severine acetate. All compounds isolated were examined for their activity against three cancer cell lines: human lung carcinoma (COR-L23), human breast adenocarcinoma (MCF7), human melanoma (C32), and normal human fetal lung cell line, MRC-5. The acridones tested exhibited weak cytotoxicity but the amides showed moderate nonselective cytotoxic activity. PMID:17365689

  3. Radiosensitization and hypoxic cell toxicity of NLA-1 and NLA-2, two new bioreductive compounds

    Energy Technology Data Exchange (ETDEWEB)

    Papadopoulou, M.V.; Epperly, M.W.; Shields, D.S. (Pittsburgh Cancer Institute PA (USA)); Bloomer, W.D.

    1992-04-01

    Two new bioreductive compounds, 9-(3-(2-nitro-1-imidazolyl)propylamino)acridine hydrochloride (NLA-1) and 9-(2-(2-nitro-1-imidazolyl)ethylamino)acridine hydrochloride (NLA-2), have been prepared. They feature an acridine ring to intercalate with DNA, a 2-nitroimidazole ring as the radiosensitizing moiety and an amino functionality for increased DNA-binding and hydrophilicity. Time concentration dependent cytotoxicity as well as radiosensitization efficacy of the two compounds under hypoxic or aerobic conditions were determined in vitro using V-79 cells and an MTT colorimetric or clonogenic assay. The isosensitization point (ISP), defined as that drug concentration which results in the same survival decrement upon exposure of hypoxic of oxygenated cells to a given radiation dose, has been determined for both compounds at 7.5 Gy and the values are significantly lower than the ISPs of 5-(3-(2-nitro-1-imidazolyl)propyl)phenanthridinium bromide, 2-(2-nitro-1-imidazolyl)ethylamine or misonidazole (MISO). NLA-1 and NLA-2 are potent hypoxic cytotoxins and on a concentration basis, more potent than MISO as radiosensitizers in vitro. The sensitization enhancement ratios were significantly increased when 1 h drug preincubation under hypoxia at 37degC was applied, before irradiation at room temperature. (author).

  4. P-Glycoprotein-Mediated Efflux and Drug Sequestration in Lysosomes Confer Advantages of K562 Multidrug Resistance Sublines to Survive Prolonged Exposure to Cytotoxic Agents

    Directory of Open Access Journals (Sweden)

    Nathupakorn Dechsupa

    2009-01-01

    Full Text Available Problem statement: Cellular drug resistance to anticancer agents is major obstacle in cancer chemotherapy and the mechanisms by which these MDR cells possess for protecting themselves to survive prolonged exposure to cytotoxic agents still debating. The study aimed to clarify the role of P-glycoprotein (Pgp and enhanced drug sequestration in lysosomes to confer the multidrug resistance K562 cells with varied degree of Pgp expression. Approach: Erythromyelogenous leukemic K562 and its corresponding Pgp-over expression K562/adr (RF = 26.5 and K562/10000 (RF = 39.6 cells were used. The transport of intrinsic fluorescence molecules including acridine orange and pirarubicin across plasma membrane of living cells was performed by using spectrofluorometric and flow cytometric analysis. Results: Pirarubicin passively diffused through the plasma membrane of K562, K562/adr and K562/10000 cells with the same values of k+ = 3.4±0.3 pL. s-1.cell-1. Similar results were found for acridine orange, which passively diffused through plasma membrane of these cell lines about 30-fold faster than pirarubicin. The mean rate of Pgp-mediated efflux coefficient (ka of pirarubicin was equal to 2.6 ± 0.9 pL.s-1.cell-1 for K562/adr and 4.7 ± 1.0 pL.s-1.cell-1 for K562/10000 cells. The Pgp-mediated efflux of acridine orange could not be determined for K562/adr cells while an enhancement of exocytosis in K562/10000 cells was characterized. The acridine orange exhibited antiproliferative activity and IC50 for K562, K562/adr and K562/10000 cells was 447±40, 715±19 and 1,719±258 nM, respectively. Cytotoxicity of acridine orange was increased by 2-fold in the presence of and 25 nM monensin. Conclusion: The results clearly demonstrated for the first time that by using the same methods and cell lines. The predominant cellular defense mechanism determined in multidrug resistant cells depends upon the nature of molecular probes used. As molecular probe, pirarubicin clearly

  5. Kinetics and mechanism of desulfurization and denitrogenation of coal-derived liquids. Tenth quarterly report, September 21-December 20, 1977

    Energy Technology Data Exchange (ETDEWEB)

    Gates, B. C.; Katzer, J. R.; Olson, J. H.; Kwart, H.; Stiles, A. B.

    1978-01-20

    Three high-pressure flow microreactors and two batch autoclave reactors have been used to study the reaction networks and kinetics of: (1) catalytic hydrodesulfurization of dibenzothiophene and methyl-substituted dibenzothiophenes; and (2) catalytic hydrodenitrogenation of quinoline, methyl-substituted quinolines, acridine and carbazole. The catalysts were commercial, sulfided CoO-MoO/sub 3//..gamma..-Al/sub 2/O/sub 3/, NiO-MoO/sub 3//..gamma..-Al/sub 2/O/sub 3/, and NiO-WO/sub 3//..gamma..-Al/sub 2/O/sub 3/. At the typical conditions of 300/sup 0/C and 104 atm, dibenzothiophene reacts to give H/sub 2/S and biphenyl in high yield, but there is some hydrogenation preceding desulfurization. Methyl-substituted dibenzothiophenes react similarly, and each reaction is first-order in the sulfur-containing compound. Two methyl groups near the sulfur atom (in the 4 and 6 positions) reduce the reactivity tenfold, whereas methyl groups in positions further removed from the sulfur atom increase reactivity about twofold. The results are consistent with steric and inductive effects influencing adsorption. The data indicate competitive adsorption among the sulfur-containing compounds. In quinoline hydrodenitrogenation, both rings are saturated before the C-N bond is broken. Similarly, in acridine conversion a large amount of hydrogenation precedes nitrogen removal. Breaking of the carbon-nitrogen bond is evidently one of the slower reactions in the network. The Ni-Mo catalyst is about twice as active as the Co-Mo catalyst for ring hydrogenation, and the two catalysts are about equally active for breaking the carbon-nitrogen bond. Reactivity of carbazole is slightly lower than that of quinoline but higher than that of acridine.

  6. Soluble Flavanthrone Derivatives: Synthesis, Characterization, and Application to Organic Light-Emitting Diodes.

    Science.gov (United States)

    Kotwica, Kamil; Bujak, Piotr; Data, Przemyslaw; Krzywiec, Wojciech; Wamil, Damian; Gunka, Piotr A; Skorka, Lukasz; Jaroch, Tomasz; Nowakowski, Robert; Pron, Adam; Monkman, Andrew

    2016-06-01

    Simple modification of benzo[h]benz[5,6]acridino[2,1,9,8-klmna]acridine-8,16-dione, an old and almost-forgotten vat dye, by reduction of its carbonyl groups and subsequent O-alkylation, yields solution-processable, electroactive, conjugated compounds of the periazaacene type, suitable for the use in organic electronics. Their electrochemically determined ionization potential and electron affinity of about 5.2 and -3.2 eV, respectively, are essentially independent of the length of the alkoxyl substituent and in good agreement with DFT calculations. The crystal structure of 8,16-dioctyloxybenzo[h]benz[5,6]acridino[2,1,9,8-klmna]acridine (FC-8), the most promising compound, was solved. It crystallizes in space group P1‾ and forms π-stacked columns held together in the 3D structure by dispersion forces, mainly between interdigitated alkyl chains. Molecules of FC-8 have a strong tendency to self-organize in monolayers deposited on a highly oriented pyrolytic graphite surface, as observed by STM. 8,16-Dialkoxybenzo[h]benz[5,6]acridino[2,1,9,8-klmna]acridines are highly luminescent, and all have photoluminescence quantum yields of about 80 %. They show efficient electroluminescence, and can be used as guest molecules with a 4,4'-bis(N-carbazolyl)-1,1'-biphenyl host in guest/host-type organic light-emitting diodes. The best fabricated diodes showed a luminance of about 1900 cd m(-12) , a luminance efficiency of about 3 cd A(-1) , and external quantum efficiencies exceeding 0.9 %.

  7. Interferon-alpha-2b induces autophagy in hepatocellular carcinoma cells through Beclin1 pathway

    International Nuclear Information System (INIS)

    To determine whether Interferon-alpha-2b (IFN-α2b) can modulate the autophagic response in hepatocellular carcinoma cells. Hepatocellular carcinoma cells were treated with IFN-α2b. Autophagy was assessed by acridine orange staining, GFP-LC3 dotted assay, transmission electron microscopy and immunoblotting. Acridine orange staining showed that IFN-α2b triggered the accumulation of acidic vesicular and autolysosomes in HepG2 cells. The acridine orange HepG2 cell ratios were (4.3±1.0)%, (6.9±1.4)%, and (13.1±2.3)%, respectively, after treatment with 100, 1,000, and 10,000 IU/mL IFN-α2b for 48 h. A markedly punctate pattern was observed in HepG2 cells treated with 10,000 IU/mL IFN-α2b for 48 h, but only diffuse and weakly fluorescent GFP-LC3 puncta was observed in control cells. HepG2 cells treated with 10,000 IU/mL IFN-α2b for 48 h developed autophagosome-like characteristics, including single- or double-membrane vacuoles containing intact and degraded cellular debris. The Beclin1 and LC3-II protein expression was up-regulated by IFN-α2b treatment. Autophagy can be induced in a dose-dependent manner by treatment with IFN-α2b in HepG2 cells, and the Beclin1 signaling pathway was stimulated by IFN-α2b

  8. Genotoxicity of heterocyclic PAHs in the micronucleus assay with the fish liver cell line RTL-W1.

    Directory of Open Access Journals (Sweden)

    Markus Brinkmann

    Full Text Available Heterocyclic aromatic hydrocarbons are, together with their un-substituted analogues, widely distributed throughout all environmental compartments. While fate and effects of homocyclic PAHs are well-understood, there are still data gaps concerning the ecotoxicology of heterocyclic PAHs: Only few publications are available investigating these substances using in vitro bioassays. Here, we present a study focusing on the identification and quantification of clastogenic and aneugenic effects in the micronucleus assay with the fish liver cell line RTL-W1 that was originally derived from rainbow trout (Oncorhynchus mykiss. Real concentrations of the test items after incubation without cells were determined to assess chemical losses due to, e.g., sorption or volatilization, by means of gas chromatography-mass spectrometry. We were able to show genotoxic effects for six compounds that have not been reported in vertebrate systems before. Out of the tested substances, 2,3-dimethylbenzofuran, benzothiophene, quinoline and 6-methylquinoline did not cause substantial induction of micronuclei in the cell line. Acridine caused the highest absolute induction. Carbazole, acridine and dibenzothiophene were the most potent substances compared with 4-nitroquinoline oxide, a well characterized genotoxicant with high potency used as standard. Dibenzofuran was positive in our investigation and tested negative before in a mammalian system. Chemical losses during incubation ranged from 29.3% (acridine to 91.7% (benzofuran and may be a confounding factor in studies without chemical analyses, leading to an underestimation of the real potency. The relative potency of the investigated substances was high compared with their un-substituted PAH analogues, only the latter being typically monitored as priority or indicator pollutants. Hetero-PAHs are widely distributed in the environment and even more mobile, e.g. in ground water, than homocyclic PAHs due to the higher water

  9. Synthesis and photophysics of a novel photocatalyst for hydrogen production based on a tetrapyridoacridine bridging ligand

    International Nuclear Information System (INIS)

    Highlights: ► Synthesis of the novel photocatalyst [(tbbpy)2Ru(tpac)PdCl2]2+RutpacPd. ► Correlation of intramolecular electron transfer with catalytic turnover. ► Reduced sensitivity to water caused by the tpac bridging-ligand. ► Dynamics in tetrapyrido[3,2-a:2′,3′-c:3″,2″-h:2‴,3‴-j]acridine complex. - Abstract: Molecular photocatalysts allow for selectively tuning their function on a molecular level based on an in-depth understanding of their chemical and photophysical properties. This contribution reports the synthesis and photophysical characterization of the novel molecular photocatalyst [(tbbpy)2Ru(tpac)PdCl2]2+RutpacPd (with tpac = tetrapyrido[3,2-a:2′,3′-c:3″,2″-h:2‴,3‴-j]acridine) and its mononuclear building block. Furthermore, detailed photocatalytic activity measurements of RutpacPd are presented. The introduction of the tpac-ligand into the molecular framework offers a potential route to reduce the impact of water as compared to the well-studied class of RutpphzPd (with tpphz = tetrapyrido[3,2-a:2′,3′-c:3″,2″-h:2‴,3‴-j]phenazine) complexes. The distinct impact of water on the electron-transfer processes in tpphz-ligands stems from the possibility of water to form hydrogen bonds to the phenazine nitrogen atoms and will potentially reduced when replacing the phenazine by the acridine unit. The effect of this structural variation on the catalytic properties and the underlying ultrafast intramolecular charge transfer behavior will be discussed in detail.

  10. Antiparasitic activities of acridone alkaloids from Swinglea glutinosa (Bl.) Merr

    Energy Technology Data Exchange (ETDEWEB)

    Santos, Djalma A.P. dos; Vieira, Paulo C.; Silva, M. Fatima das G.F. da; Fernandes, Joao B. [Universidade Federal de Sao Carlos, SP (Brazil). Dept. de Quimica; Rattray, Lauren; Croft, Simon L. [London School of Hygiene and Tropical Medicine, London (United Kingdom). Dept. of Infectious and Tropical Diseases

    2009-07-01

    Eleven acridone alkaloids isolated from Swinglea glutinosa (Bl.) Merr. were examined for in vitro activity against chloroquine-sensitive Plasmodium falciparum 3D7, Trypanosoma brucei rhodesiense STIB900 and Leishmania donovani L82. An assay with KB cells was developed in order to compare in vitro toxicity of alkaloids with the selective action on the parasites. Nine of the compounds had IC{sub 50} values ranging from 0.3 to 11.6 {mu}M against P. falciparum. In contrast, a small number of compounds showed significant activity against T. brucei rhodesiense and none had activity against L. donovani. Among the alkaloids three had IC{sub 50} < 1.0 {mu}M against P. falciparum, whereas against T. b. rhodesiense five had IC{sub 50} < 10 {mu}M. The characterization of the acridone alkaloids, 1,3,5-trihydroxy-4-methoxy-10-methyl-2,8-bis(3-methylbut-2-enyl)acridin-9 (10H)-one (1), 2,3-dihydro-4,9-dihydroxy-2-(2-hydroxypropan-2-yl)-11-methoxy-10-methylfuro [3,2-b] acridin-5(10H)-one (2) and 3,4-dihydro-3,5,8-trihydroxy-6-methoxy-2,2,7-trimethyl-2Hpyrano[ 2,3-a]acridin-12(7H)-one (3), is discussed, as well as the structure-activity relationship of all compounds assayed. Isolation and spectral data of alkaloids 1-3 are described for the first time although their cytotoxicities to cancer cells have been described before. (author)

  11. Photosensitized oxidation of DNA and its components

    International Nuclear Information System (INIS)

    Chemical changes in DNA components during the photodynamic effect are responsible for Mutagenic and carcinogenic phenomena. Basically two competitive mechanisns involving respectively a charge transfer (type I) and singlet oxygen (type II) are implicated in reactions photo-sensitized by different agents (acridines, phenothiazines, porphyrins, flavins, psoralenes...). A study of the photosensitized oxidation of DNA itself was approached through characterization of the main final products in the case of purine nucleosides. Methyl-2 naphthoquinone - 1,4 (vitamin K3) displays a special photosensitization mechanism involving a cation radical type of intermediary

  12. Usefulness of quantitative buffy coat blood parasite detection system in diagnosis of malaria

    OpenAIRE

    Pinto M; Rodrigues S; Desouza R; Verenkar M

    2001-01-01

    A rapid test for diagnosis of malaria based on acridine orange staining of centrifuged blood samples in a microhematocrit tube (QBC) was compared with thick and thin peripheral blood smears in 2274 samples. Malaria was diagnosed in 239 (10.5%) patients by Leishman′s staining technique and QBC method. The QBC method allowed detection of an additional 89 (3.9%) cases. Thus the prevalence rate of malaria during the study was 14.4%. In 1946 patients who were negative b...

  13. Synthesis and In Vitro Testing of New Potent Polyacridine-Melittin Gene Delivery Peptides

    OpenAIRE

    Baumhover, Nicholas J.; Anderson, Kevin; Fernandez, Christian A.; Rice, Kevin G.

    2010-01-01

    The combination of a polyacridine peptide modified with a melittin fusogenic peptide results in a potent gene transfer agent. Polyacridine peptides of the general formula (Acr-X)n-Cys were prepared by solid phase peptide synthesis, where Acr is Lys modified on its ε-amine with acridine, X is Arg, Leu or Lys and n is 2, 3 or 4 repeats. The Cys residue was modified by either a maleimide-melittin or a thiolpyridine-Cys-melittin fusogenic peptide resulting in reducible or non-reducible polyacridi...

  14. Photocatalytic performance of TiO2 immobilized on polystyrene (PS) thin film for mineralization of pollutants

    Institute of Scientific and Technical Information of China (English)

    2003-01-01

    A new photocatalyst, TiO2 powder immobilized on polystyrene (PS) thin films, was prepared using a novelmethod and its photocatalytic activity on the photodegradation of acridine dye in aqueous solution was tested. By thismethod, the crystal form and grain size of the immobilized TiO2 were well maintained. Compared with TiO2 powder, thephotocatalytic activity of TiO2/PS thin films was not significantly reduced. The catalyst is stable and can be reused severaltimes without the loss of activity, which makes wastewater treatment using this photocatalytic degradation technique of thisway possible in the practical application.

  15. Cell volume and geometric parameters determination in living cells using confocal microscopy and 3D reconstruction

    OpenAIRE

    sprotocols

    2015-01-01

    Authors: David Hevia, Aida Rodriguez-Garcia, Marta Alonso-Gervós, Isabel Quirós-González, Henar M Cimadevilla, Carmen Gómez-Cordovés, Rosa M Sainz & Juan C Mayo ### Abstract The protocol reported here describes a simple, easy, fast and reproducible method aimed to know the geometric parameters of living cells based on confocal laser scanning microscopy combined with 3D reconstruction software. Briefly, the method is based on intrinsic fluorescence properties of acridine orange (AO...

  16. Evaluation of sperm chromatin structure in boar semen

    Directory of Open Access Journals (Sweden)

    Banaszewska Dorota

    2015-06-01

    Full Text Available This study was an attempt to evaluate sperm chromatin structure in the semen of insemination boars. Preparations of semen were stained with acridine orange, aniline blue, and chromomycin A3. Abnormal protamination occurred more frequently in young individuals whose sexual development was not yet complete, but may also be an individual trait. This possibility is important to factor into the decision regarding further exploitation of insemination boars. Thus a precise assessment of abnormalities in the protamination process would seem to be expedient as a tool supplementing morphological and molecular evaluation of semen. Disruptions in nucleoprotein structure can be treated as indicators of the biological value of sperm cells.

  17. Efficacy of Sex Determination from Human Dental Pulp Tissue and its Reliability as a Tool in Forensic Dentistry

    OpenAIRE

    Khanna, Kaveri Surya

    2015-01-01

    Background: Sex determination is one of the primary steps in forensics. Barr body can be used as a histological method for identification of sex as it is found to be specific to female somatic cells and rare in male cells. To demarcate human dental pulp as an important identification tool of sex in forensic odontology (FO) and to evaluate the time period till which sex can be determined from pulp tissue using three stains H and E, Feulgen, and acridine - orange under fluorescence so as. Mater...

  18. Comparison of Herpes simplex virus plaque development after viral treatment with anti-DNA or antilipid agents

    Energy Technology Data Exchange (ETDEWEB)

    Coohill, T.P.; Babich, M.; Taylor, W.D.; Snipes, W.

    1980-06-01

    The plaque development of Herpes simplex virus type 1 (HSV) is slower for viruses treated with two anti-DNA agents: ultraviolet radiation (uv) or n-acetoxy-2-acetyl-aminofluorene. For HSV treated with three antimembrane agents - butylated hydroxytoluene, acridine plus near uv radiation, or ether - the plaque development time is the same as for untreated viruses. These differences hold even for viruses that survived treatment that lowered viability below the 1% level. Gamma ray inactivation of HSV produces no change in plaque development even though this agent is believed to preferentially affect viral DNA.

  19. IMAGING WOOD PULP FIBRE SURFACE LIGNIN BY FLUORESCENCE CONFOCAL LASER SCANNING MICROSCOPY

    Institute of Scientific and Technical Information of China (English)

    KechengLi; DouglasW.Reeve

    2004-01-01

    A novel methodology for imaging wood pulp fibre surface lignin by fluorescence confocal laser scanning microscopy was developed. Various imaging modes and imaging conditions were explored for quantitative analysis. Acridine Orange was used for labelling lignin and the orthochromatic labelling condition was developed. With the thusly established methodology, the distribution of lignin across the fibre wall was clearly imaged. It was found that surface lignin concentration is about 2-4 times higher than bulk lignin concentration and that high concentration of lignin was also found on the fibre lumen surfaces and pit borders.

  20. IMAGING WOOD PULP FIBRE SURFACE LIGNIN BY FLUORESCENCE CONFOCAL LASER SCANNING MICROSCOPY

    Institute of Scientific and Technical Information of China (English)

    Kecheng Li; Douglas W. Reeve

    2004-01-01

    A novel methodology for imaging wood pulp fibre surface lignin by fluorescence confocal laser scanning microscopy was developed. Various imaging modes and imaging conditions were explored for quantitative analysis. Acridine Orange was used for labelling lignin and the orthochromatic labelling condition was developed. Withthe thusly established methodology, the distribution of lignin across the fibre wall was clearly imaged. It was found that surface lignin concentration is about 2-4 times higher than bulk lignin concentration, and that high concentration of lignin was also found on the fibre lumen surfaces and pit borders.

  1. Dual AO/EB Staining to Detect Apoptosis in Osteosarcoma Cells Compared with Flow Cytometry

    OpenAIRE

    Liu, Kuan; Liu, Peng-Cheng; Liu, Run; Wu, Xing

    2015-01-01

    Background The aim of this study was to evaluate the ability of dual acridine orange/ethidium bromide (AO/EB) staining to detect tumor cell apoptosis. According to apoptosis-associated changes of cell membranes during the process of apoptosis, a clear distinction is made between normal cells, early and late apoptotic cells, and necrotic cells. Material/Method We cultured human osteosarcoma cells with 30, 60, and 120 μg/ml kappa-selenocarrageenan. To assess the rates of cell proliferation and ...

  2. Laboratory simulation of meteoritic noble gases. III - Sorption of neon, argon, krypton, and xenon on carbon - Elemental fractionation

    Science.gov (United States)

    Wacker, John F.

    1989-01-01

    The sorption of Ne, Ar, Kr, and Xe was studied in carbon black, acridine carbon, and diamond in an attempt to understand the origin of trapped noble gases in meteorites. The results support a model in which gases are physically adsorbed on interior surfaces formed by a pore labyrinth within amorphous carbons. The data show that: (1) the adsorption/desorption times are controlled by choke points that restrict the movement of noble gas atoms within the pore labyrinth, and (2) the physical adsorption controls the temperature behavior and elemental fractionation patterns.

  3. Microwave-prompted Reaction of Cinnamonitrile Derivatives with 5,5-Dimethyl-1,3-cyclohexanedione

    Institute of Scientific and Technical Information of China (English)

    TU,Shu-Jiang(屠树江); MIAO,Chun-Bao(缪春宝); GAO,Yuan(高原); FENG,You-Jian(冯友健); FENG,Jun-Cai(冯骏材)

    2002-01-01

    In the reactions of α-cyanocinamonitrile or β-cyano-β-carbo-thoxy styrene with 5,5-dimethyl-1,3-cyclohexanedione in the presence of ammonium acetate under microwave irradiation without solvent, the 2-amino-5,6,7,8-tetrahydro-5-oxo-4-aryl-7,7-dimethyl-4H-benzo-[b]-pyran derivatives were obtained.However, in the reasons of arylidenecyanoacetamide with 5,5-dimethyl-1,3-cyclohexanedione under the same reaction conditions, the acridine derivatives were obtained. The structures of the products were determined by single crystal X-ray diffraction analysis.

  4. Monocyte functions in diabetes mellitus.

    Science.gov (United States)

    Geisler, C; Almdal, T; Bennedsen, J; Rhodes, J M; Kølendorf, K

    1982-02-01

    The aim of this study was to investigate the functions of monocytes obtained from 14 patients with diabetes mellitus (DM) compared with those of monocytes from healthy individuals. It was found that the total number of circulating monocytes in the 14 diabetic patients was lower than that from the healthy individuals. Phagocytosis of Candida albicans was decreased in the monocytes from the patients, whereas pinocytosis of acridine and phagocytosis of latex and sheep red blood cells were normal. The chemotactic response towards casein was enhanced. The possible consequences of these findings for the elucidation of concomitant infections in diabetic patients are discussed.

  5. Marine aggregates and transparent exopolymeric particles (TEPs) as substrates for the stramenopilan fungi, the thraustochytrids: Roller table experimental approach.

    Digital Repository Service at National Institute of Oceanography (India)

    Damare, V.S.; Raghukumar, S.

    . The samples were preserved with formaldehyde at a final concentration of 2 % and stored at 5o C. Total bacteria, thraustochytrids and TEPs were analyzed. Ten mL of water was filtered over 25 mm, 0.22 m black polycarbonate Isopore membrane filters (Millipore..., USA). The filter paper was stained for bacteria by the acridine orange direct count (AODC) method (Parsons et al., 1984). Thraustochytrids were enumerated by filtering 25 mL of water over 0.4 m black polycarbonate Isopore membrane filter (Millipore...

  6. “Human Babesiosis”: An Emerging Transfusion Dilemma

    Directory of Open Access Journals (Sweden)

    Helieh S. Oz

    2012-01-01

    Full Text Available Babesiosis, a common disease of animals, can infect humans via vector “tick bite”, particularly in endemic areas. The recent reports of fatal cases in Hepatitis C and postliver transplant patients resulting from transfusion of contaminated blood should alert the medical profession regarding this emerging dilemma in endemic as well as nonendemic areas and the need for accurate blood screening for transfusion. Here, we illustrate different stages of the parasite lifecycle, progression of babesiosis in animal model, some aspects of pathologic outcomes, ongoing therapeutic modalities, and a feasible Acridine Orange fluorescent methodology for the diagnostic evaluation of blood samples.

  7. 不同荧光染料对苜蓿原生质体染色效果的研究%Effects of Different Fluorescent Dyes on Staining Alfalfa Protoplast

    Institute of Scientific and Technical Information of China (English)

    李玉珠; 师尚礼; 贺延玉

    2012-01-01

    The effects of five fluorescent dyes (FDA, Rhodamine B, Rhodamine 6G, Acridine Orange and DAPI) on staining protoplasts obtained from callus of Madicago sativa ' Gannong No. 4' were investigated in order to test the viability of protoplasts, count nuclei and survey processes of protoplast fusion. Result showed that FDA, Rhodamine B, Rhodamine 6G and Acridine Orange dye and mark protoplasts of alfalfa clearly, which could identify parental protoplasts from fusion clusters. FDA is an optimal dye for testing the viability of protoplasts. Acridine Orange and DAPI can dye specific nucleus of protoplasts, which are then used to count and survey nuclei in cell fusion. Acridine Orange can also test the viability of fusion cells. These studies suggest a basis for exploring cell fusion conditions and culturing fusion cells by identifying heterocaryon and testing their viability.%以紫花苜蓿品种甘农4号(Madicago sativa L.‘Gannong No.4’)愈伤组织酶解的原生质体为材料,采用FDA、罗丹明B、罗丹明6G、吖啶橙和DAPI共5种荧光染料对原生质体进行染色效果比较,以期为原生质体活力测定、细胞核计数及原生质体融合过程的观察提供参考.结果表明:FDA、罗丹明B、罗丹明6G和吖啶橙均可使紫花苜蓿原生质体清晰染色,可用于细胞融合时亲本原生质体的标识.FDA是检测原生质体活力的理想染料;吖啶橙和DAPI可对细胞核进行特异性染色,可用于融合时细胞核的观察和计数,前者还可用于检测融合细胞的活力.通过对异源融合体的鉴别及其活力的测定,为摸索融合条件和融合细胞的培养提供了理论依据.

  8. Integrative biological studies of anti-tumour agents

    OpenAIRE

    Johnson, L. A.

    2009-01-01

    3, 11-difluoro-6, 8, 13-trimethyl-8H- quino [4, 3, 2-kl] acridinium methosulfate (RHPS4) is a member of a series of pentacyclic acridines developed at the University of Nottingham, which bind to, and stabilise the structure of G-quadruplex DNA and inhibit the action of telomerase at sub-micromolar concentrations in the cell free TRAP assay and limit cancer cell growth therefore leading to the conclusion that RHPS4 has potential anti-tumour activity. Previous biological studies, however, have...

  9. Role of vacuolar membrane proton pumps in the acidification of protein storage vacuoles following germination.

    Science.gov (United States)

    Wilson, Karl A; Chavda, Burzin J; Pierre-Louis, Gandhy; Quinn, Adam; Tan-Wilson, Anna

    2016-07-01

    During soybean (Glycine max (L.) Merrill) seed development, protease C1, the proteolytic enzyme that initiates breakdown of the storage globulins β-conglycinin and glycinin at acidic pH, is present in the protein storage vacuoles (PSVs), the same subcellular compartments in seed cotyledons where its protein substrates accumulate. Actual proteolysis begins to be evident 24 h after seed imbibition, when the PSVs become acidic, as indicated by acridine orange accumulation visualized by confocal microscopy. Imidodiphosphate (IDP), a non-hydrolyzable substrate analog of proton-translocating pyrophosphatases, strongly inhibited acidification of the PSVs in the cotyledons. Consistent with this finding, IDP treatment inhibited mobilization of β-conglycinin and glycinin, the inhibition being greater at 3 days compared to 6 days after seed imbibition. The embryonic axis does not appear to play a role in the initial PSV acidification in the cotyledon, as axis detachment did not prevent acridine orange accumulation three days after imbibition. SDS-PAGE and immunoblot analyses of cotyledon protein extracts were consistent with limited digestion of the 7S and 11S globulins by protease C1 starting at the same time and proceeding at the same rate in detached cotyledons compared to cotyledons of intact seedlings. Embryonic axis removal did slow down further breakdown of the storage globulins by reactions known to be catalyzed by protease C2, a cysteine protease that normally appears later in seedling growth to continue the storage protein breakdown initiated by protease C1. PMID:27043965

  10. Use of the comet assay to measure DNA damage in cells exposed to photosensitizers and gamma radiation

    Science.gov (United States)

    Pouget, J.-P.; Ravanat, J.-L.; Douki, T.; Richard, M.-J.; Cadet, J.

    1999-01-01

    We used the comet assay associated with DNA-glycosylases to estimate DNA damage in cells exposed to gamma irradiation or photosensitized either with methylene blue or orange acridine. A calibration performed using irradiation allowed the measurement of the steady-state level and the yield of 8-oxodGuo as well as strand breaks and alkali-labile sites. Nous avons utilisé la méthode des comètes associée à des ADN-glycosylases, pour estimer les dommages de l'ADN dans des cellules après l'exposition à un rayonnement gamma ou après photosensibilisation par le bleu de méthylène ou l'acridine orange. Une calibration de la méthode des comètes a permis de mesurer le niveau basal et les taux de formation de 8-oxodGuo ainsi que le nombre de cassures de brins et de sites alcali labiles.

  11. Comparison between morphological and staining characteristics of live and dead eggs of Schistosoma mansoni

    Directory of Open Access Journals (Sweden)

    AK Sarvel

    2006-10-01

    Full Text Available Schistosoma mansoni eggs are classified, according to morphological characteristics, as follows: viable mature and immature eggs; dead mature and immature eggs, shells and granulomas. The scope of this study was to compare the staining characteristics of different morphological types of eggs in the presence of fluorescent labels and vital dyes, aiming at differentiating live and dead eggs. The eggs were obtained from the intestines of infected mice, and put into saline 0.85%. The fluorescent labels were Hoechst 33258 and Acridine Orange + Ethidium Bromide and vital dyes (Trypan Blue 0.4% and Neutral Red 1%. When labelled with the probe Hoechst 33258, some immature eggs, morphologically considered viable, presented fluorescence (a staining characteristic detected only in dead eggs; mature eggs did not present fluorescence, and the other types of dead eggs, morphologically defined, showed fluorescence. As far as Acridine Orange + Ethidium Bromide are concerned, either the eggs considered to be live, or the dead ones, presented staining with green color, and only the hatched and motionless miracidium was stained with an orange color. Trypan Blue was not able to stain the eggs, considered to be dead but only dead miracidia which had emerged out of the shell. Neutral Red stained both live and dead eggs. Only the fluorescent Hoechst 33258 can be considered a useful tool for differentiation between dead and live eggs.

  12. Integrated scientific data bases review on asulacrine and associated toxicity.

    Science.gov (United States)

    Afzal, Attia; Sarfraz, Muhammad; Wu, Zimei; Wang, Guangji; Sun, Jianguo

    2016-08-01

    Asulacrine (ASL), a weakly basic and highly lipophilic drug was synthesized in 1980's in cancer research laboratory of Auckland by modifications to the acridine portion of amsacrine on 3-, 4- and 5-substitution patterns. In contrast to its precursor amsacrine (m-AMSA), ASL was effective not only against leukemia and Lewis lung tumor system but also a wide variety of solid tumor. Its metabolic pathway is not same to amsacrine hence different side effects, hepatotoxicity and excretion was observed. Asulacrine is under phase II clinical trials and has showed promising results but its toxicity especially phlebitis is stumbling block in its clinical implementation. This review is an effort to give a possible clue, based on scientifically proven results, to the researchers to solve the mystery of associated toxicity, phlebitis. Review covers the available literature on asulacrine and other acridine derivatives regarding pharmacology, pharmacokinetics, quantitative structure activity relationship and toxicology via electronic search using scientific databases like PubMed and others. To date, all abstracts and full-text articles were discussed and analyzed. The tabulated comparisons and circuitry mechanism of ASL are the added features of the review which give a complete understanding of hidden aspects of possible route cause of associated toxicity, the phlebitis.

  13. A comparative study of quantitative stains for DNA in image cytometry.

    Science.gov (United States)

    Mikel, U V; Becker, R L

    1991-08-01

    In this study we examined the reproducibility of several stains used to measure nuclear DNA by image cytometry. The specimens were touch preparations of liver and testis from mouse and liver, intestine and brain from rat, fixed in either neutral formalin or Carnoy's solution. The tested stains included four Feulgen methods (pararosaniline, azure-A, thionin and acriflavine), the gallocyanine-chromalum stain and two fluorescent stains (acridine orange and propidium iodide). Absorbance measurements employed a video image analysis system; fluorescence measurements were from a scanning microspectrophotometer. The acriflavine-Feulgen stain was analyzed for both absorbance and fluorescence. All seven stains were quantitative for DNA and gave reproducible results. The absorbance measurements had a lower coefficient of variation (CV) than the fluorescence values. In a nested analysis of variance of the pararosaniline Feulgen stains, cell-to-cell variability accounted for 67% of the total variance; slide-to-slide, 9%; and batch-to-batch, 24%. These values did not change significantly when the staining was performed in an automatic staining machine. For DNA analysis using image cytometry, we conclude that the Feulgen staining technique is the most useful. In particular, acriflavine-Feulgen-stained cells fixed in Carnoy's fluid give the least variation between measurement values and the most accurate ratios between the separate ploidy groups. For fluorescence cytometry we recommend Carnoy's fixation and the acriflavine-Feulgen stain because of its narrow CV as compared to acridine orange and propidium iodide. PMID:1718295

  14. Acridone Alkaloids from Swinglea glutinosa (Rutaceae) and Their Effects on Photosynthesis.

    Science.gov (United States)

    Arato Ferreira, Pedro H; Dos Santos, Djalma A P; da Silva, Maria Fátima das G F; Vieira, Paulo C; King-Diaz, Beatriz; Lotina-Hennsen, Blas; Veiga, Thiago A M

    2016-01-01

    Continuing our search for herbicide models based on natural products, we investigated the action mechanisms of five alkaloids isolated from Swinglea glutinosa (Rutaceae): Citrusinine-I (1), glycocitrine-IV (2), 1,3,5-trihydroxy-10-methyl- 2,8-bis(3-methylbut-2-en-1-yl)-9(10H)-acridinone (3), (2R)-2-tert-butyl-3,10-dihydro-4,9-dihydroxy-11-methoxy-10-methylfuro[3,2-b]acridin-5(2H)-one (4), and (3R)-2,3,4,7-tetrahydro-3,5,8-trihydroxy-6-methoxy-2,2,7-trimethyl-12H-pyrano[2,3-a]acridin-12-one (5) on several photosynthetic activities in an attempt to find new compounds that affect photosynthesis. Through polarographic techniques, the compounds inhibited the non-cyclic electron transport in the basal, phosphorylating, and uncoupled conditions from H2 O to methylviologen (=MV). Therefore, they act as Hill reaction inhibitors. This approach still suggested that the compounds 4 and 5 had their interaction site located at photosystem I. Studies on fluorescence of chlorophyll a suggested that acridones (1-3) have different modes of interaction and inhibition sites on the photosystem II electron transport chain. PMID:26765357

  15. Genotoxicity of non-covalent interactions: DNA intercalators

    Energy Technology Data Exchange (ETDEWEB)

    Ferguson, Lynnette R. [Auckland Cancer Society Research Centre, Faculty of Medical and Health Science, University of Auckland (New Zealand)], E-mail: l.ferguson@auckland.ac.nz; Denny, William A. [Auckland Cancer Society Research Centre, Faculty of Medical and Health Science, University of Auckland (New Zealand)

    2007-10-01

    This review provides an update on the mutagenicity of intercalating chemicals, as carried out over the last 17 years. The most extensively studied DNA intercalating agents are acridine and its derivatives, that bind reversibly but non-covalently to DNA. These are frameshift mutagens, especially in bacteria and bacteriophage, but do not otherwise show a wide range of mutagenic properties. Di-acridines or di-quinolines may be either mono- or bis-intercalators, depending upon the length of the alkyl chain separating the chromophores. Those which monointercalate appear as either weak frameshift mutagens in bacteria, or as non-mutagens. However, some of the bisintercalators act as 'petite' mutagens in Saccharomyces cerevisiae, suggesting that they may be more likely to target mitochondrial as compared with nuclear DNA. Some of the new methodologies for detecting intercalation suggest this may be a property of a wider range of chemicals than previously recognised. For example, quite a number of flavonoids appear to intercalate into DNA. However, their mutagenic properties may be dominated by the fact that many of them are also able to inhibit topoisomerase II enzymes, and this property implies that they will be potent recombinogens and clastogens. DNA intercalation may serve to position other, chemically reactive molecules, in specific ways on the DNA, leading to a distinctive (and wider) range of mutagenic properties, and possible carcinogenic potential.

  16. Induction of apoptosis on human hepatocarcinoma cell lines by an alkyl resorcinol isolated from Lithraea molleoides

    Institute of Scientific and Technical Information of China (English)

    Luciana Barbini; Paula Lopez; Julieta Ruffa; Virginia Martino; Graciela Ferraro; Rodolfo Campos; Lucia Cavallaro

    2006-01-01

    AIM: To study the mechanism of cytotoxicity of a new active 5-alkyl resorcinol [1, 3-dihydroxy-5- (tridec-4', 7'-dienyl) benzene] isolated from Lithraea molleoides leaves on liver tumor cells.METHODS: Human hepatocarcinoma cell lines (HepG2and Hep3B) in culture were treated with inhibitory concentrations, 50% of the compound, for 24 h. The induction of apoptosis was detected in treated cells by analysis of DNA fragmentation, DNA content, and acridine orange and propidium iodide staining.RESULTS: After 24 h of 5-alkyl resorcinol treatment,both cell lines showed: (1) the typical morphological alterations of apoptosis; (2) DNA fragmentation, detected by laddering and appearance of a subG0 population by flow cytometry; and (3) condensed and fragmented nuclei by acridine orange-propidium iodide staining.CONCLUSION: Based on the results, this compound exerts its cytotoxic effect in both hepatocellular cell lines through apoptotic cell death. For Hep3B, cells with mutated p53 and Fas, apoptosis would proceed by p53-or Fas-independent pathways.

  17. Groundwater contamination by organic bases derived from coal-tar wastes

    Science.gov (United States)

    Pereira, W.E.; Rostad, C.E.; Garbarino, J.R.; Hult, M.F.

    1983-01-01

    A fluid sample from a shallow aquifer contaminated by coal-tar wastes was analyzed for organic bases. The sample consisted of a mixture of aqueous and oily-tar phases. The phases were separated by centrifugation and filtration. Organic bases were isolated from each phase by pH adjustment and solvent extraction. Organic bases in the oily-tar phase were further purified by neutral-alumina, micro-column adsorption chromatography. Separation and identification of the organic bases in each phase were achieved by using capillary gas chromatography-mass spectrometry-computer (GC-MS-COM) and probe distillation-high resolution mass spectrometry (PD-HRMS) techniques. Organic bases present in the aqueous phase included primary aromatic amines (such as aniline, alkylated anilines, and naphthylamines) as well as azaarenes (such as alkylated pyridines, quinolines, acridine, and benzoquinolines). The oily-tar phase contained acridine, benzacridines, dibenzacridines, and numerous other azaarenes, the elemental compositions of which were determined by PD-HRMS. Azaarenes in the oily-tar phase, varying in size from 6 to 12 rings, are reported for the first time. The origin and environmental significance of these compounds are discussed. ?? 1983.

  18. Ground-water contamination by organic bases derived from coal-tar wastes

    Science.gov (United States)

    Pereira, Wilfred E.; Rostad, Colleen E.; Garbarino, John R.; Hult, Marc F.

    1983-01-01

    A fluid sample from a shallow aquifer contaminated by coal-tar wastes was analyzed for organic bases. The sample consisted of a mixture of aqueous and oily-tar phases. The phases were separated by centrifugation and filtration. Organic bases were isolated from each phase by pH adjustment and solvent extraction. Organic bases in the oily-tar phase were further purified by neutral-alumina, micro-column adsorption chromatography. Separation and identification of the organic bases in each phase were achieved by using capillary gas chromatography-mass spectrometry-computer (GC-MS-COM) and probe distillation-high resolution mass spectrometry (PD-HRMS) techniques. Organic bases present in the aqueous phase included primary aromatic amines (such as aniline, alkylated anilines, and naphthylamines) as well as azaarenes (such as alkylated pyridines, quinolines, acridine, and benzoquinolines). The oily-tar phase contained acridine, benzacridines, dibenzacridines, and numerous other azaarenes, the elemental compositions of which were determined by PD-HRMS. Azaarenes in the oily-tar phase, varying in size from 6 to 12 rings, are reported for the first time. The origin and environmental significance of these compounds are discussed.

  19. An improved layer-by-layer self-assembly technique to generate biointerfaces for platelet adhesion studies: Dynamic LbL

    Science.gov (United States)

    Lopez, Juan Manuel

    Layer-by-layer self-assembly (LbL) is a technique that generates engineered nano-scale films, coatings, and particles. These nanoscale films have recently been used in multiple biomedical applications. Concurrently, microfabrication methods and advances in microfluidics are being developed and combined to create "Lab-on-a-Chip" technologies. The potential to perform complex biological assays in vitro as a first-line screening technique before moving on to animal models has made the concept of lab on a chip a valuable research tool. Prior studies in the Biofluids Laboratory at Louisiana Tech have used layer-by-layer and in vitro biological assays to study thrombogenesis in a controlled, repeatable, engineered environment. The reliability of these previously established techniques was unsatisfactory for more complex cases such as chemical and shear stress interactions. The work presented in this dissertation was performed to test the principal assumptions behind the established laboratory methodologies, suggest improvements where needed, and test the impact of these improvements on accuracy and repeatability. The assumptions to be tested were: (1) The fluorescence microscopy (FM) images of acridine orange-tagged platelets accurately provide a measure of percent area of surface covered by platelets; (2) fibrinogen coatings can be accurately controlled, interact with platelets, and do not interfere with the ability to quantify platelet adhesion; and (3) the dependence of platelet adhesion on chemical agents, as measured with the modified methods, generally agrees with results obtained from our previous methods and with known responses of platelets that have been documented in the literature. The distribution of fibrinogen on the final LbL surface generated with the standard, static process (s-LbL) was imaged by tagging the fibrinogen with an anti-fibrinogen antibody bound to fluorescein isothiocyanate (FITC). FITC FM images and acridine orange FM images were taken

  20. Betulinic Acid Inhibits Growth of Cultured Vascular Smooth Muscle Cells In Vitro by Inducing G1 Arrest and Apoptosis

    Directory of Open Access Journals (Sweden)

    Raja Kumar Vadivelu

    2012-01-01

    Full Text Available Betulinic acid is a widely available plant-derived triterpene which is reported to possess selective cytotoxic activity against cancer cells of neuroectodermal origin and leukemia. However, the potential of betulinic acid as an antiproliferative and cytotoxic agent on vascular smooth muscle (VSMC is still unclear. This study was carried out to demonstrate the antiproliferative and cytotoxic effect of betulinic acid on VSMCs using 3-[4,5-dimethylthizol-2-yl]-2,5-diphenyltetrazolium bromide (MTT assay, flow cytometry cell cycle assay, BrdU proliferation assay, acridine orange/propidium iodide staining, and comet assay. Result from MTT and BrdU assays indicated that betulinic acid was able to inhibit the growth and proliferation of VSMCs in a dose-dependent manner with IC50 of 3.8 μg/mL significantly (P<0.05. Nevertheless, betulinic acid exhibited G1 cell cycle arrest in flow cytometry cell cycle profiling and low level of DNA damage against VSMC in acridine orange/propidium iodide and comet assay after 24 h of treatment. In conclusion, betulinic acid induced G1 cell cycle arrest and dose-dependent DNA damage on VSMC.

  1. Experimental and theoretical investigation effect of flavonols antioxidants on DNA damage.

    Science.gov (United States)

    Ensafi, Ali A; Heydari-Soureshjani, E; Jafari-Asl, M; Rezaei, B; Ghasemi, Jahan B; Aghaee, Elham

    2015-08-01

    A new electrochemical biosensor was developed to demonstrate the effect of Acridine Orange (AO) on DNA damage. Then, the biosensor was used to check the inhibitors effect of three flavonols antioxidants (myricetin, fisetin and kaempferol) on DNA damage. Acridine Orange (AO) was used as a damaging agent because it shows a high affinity to nucleic acid and stretch of the double helical structure of DNA. Decreasing on the oxidation signals of adenine and guanine (in the DNA) in the presence of AO were used as probes to study the antioxidants power, using DNA-modified screen printed graphene electrode (DNA/SPGE). The results of our study showed that the DNA-biosensor could be suitable biosensor to investigate the inhibitors ability of the flavonols antioxidants on the DNA damage. The linear dependency was detected in the two regions in the ranges of 1.0-15.0 and 15.0-500.0 pmol L(-1). The detection limit was found 0.5 pmol L(-1) and 0.6 pmol L(-1) for guanine and adenine, respectively. To confirm the electrochemical results, Uv-Vis and fluorescence spectroscopic methods were used too. Finally molecular dynamic (MD) simulation was performed on the structure of DNA in a water box to study any interaction between the antioxidant, AO and DNA.

  2. Genotoxicity of non-covalent interactions: DNA intercalators

    International Nuclear Information System (INIS)

    This review provides an update on the mutagenicity of intercalating chemicals, as carried out over the last 17 years. The most extensively studied DNA intercalating agents are acridine and its derivatives, that bind reversibly but non-covalently to DNA. These are frameshift mutagens, especially in bacteria and bacteriophage, but do not otherwise show a wide range of mutagenic properties. Di-acridines or di-quinolines may be either mono- or bis-intercalators, depending upon the length of the alkyl chain separating the chromophores. Those which monointercalate appear as either weak frameshift mutagens in bacteria, or as non-mutagens. However, some of the bisintercalators act as 'petite' mutagens in Saccharomyces cerevisiae, suggesting that they may be more likely to target mitochondrial as compared with nuclear DNA. Some of the new methodologies for detecting intercalation suggest this may be a property of a wider range of chemicals than previously recognised. For example, quite a number of flavonoids appear to intercalate into DNA. However, their mutagenic properties may be dominated by the fact that many of them are also able to inhibit topoisomerase II enzymes, and this property implies that they will be potent recombinogens and clastogens. DNA intercalation may serve to position other, chemically reactive molecules, in specific ways on the DNA, leading to a distinctive (and wider) range of mutagenic properties, and possible carcinogenic potential

  3. Introduction of biocides into clinical practice and the impact on antibiotic-resistant bacteria.

    Science.gov (United States)

    Russell, A D

    2002-01-01

    Biocides and other antimicrobial agents have been employed for centuries. Much later, iodine found use as a wound disinfectant, chlorine water in obstetrics, alcohol as a hand disinfectant and phenol as a wound dressing and in antiseptic surgery. In the early part of the twentieth century, other chlorine-releasing agents (CRAs), and acridine and other dyes were introduced, as were some quaternary ammonium compounds (QACs, although these were only used as biocides from the 1930s). Later still, various phenolics and alcohols, formaldehyde and hydrogen peroxide were introduced and subsequently (although some had actually been produced at an earlier date) biguanides, iodophors, bisphenols, aldehydes, diamidines, isocyanurates, isothiazolones and peracetic acid. Antibiotics were introduced clinically in the 1940s, although sulphonamides had been synthesized and used previously. After penicillin came streptomycin and other aminoglycosides-aminocyclitols, tetracyclines, chloramphenicol, macrolides, semi-synthetic beta-lactams, glycopeptides, lincosamides, 4-quinolones and diaminopyrimidines. Bacterial resistance to antibiotics is causing great concern. Mechanisms of such resistance include cell impermeability, target site mutation, drug inactivation and drug efflux. Bacterial resistance to biocides was described in the 1950s and 1960s and is also apparently increasing. Of the biocides listed above, cationic agents (QACs, chlorhexidine, diamidines, acridines) and triclosan have been implicated as possible causes for the selection and persistence of bacterial strains with low-level antibiotic resistance. It has been claimed that the chronological emergence of qacA and qacB determinants in clinical isolates of Staphylococcus aureus mirrors the introduction and usage of cationic biocides.

  4. Single crystal X-ray diffraction studies of DNA and DNA-drug complexes

    CERN Document Server

    Todd, A K

    1999-01-01

    The structure of the brominated oligonucleotide d(ACGTACG(5-BrU)) sub 2 was solved using the multiwavelength anomalous diffraction (MAD) technique. The space group was P4 sub 3 2 sub 1 2, with unit cell a=b=43.60A, c=26.27A. This structure was an A-DNA, isomorphous with many other previously solved octomers. Single crystal X-ray diffraction data were collected from crystals of the intercalation complexes N-[2-(dimethylamino)ethyl] acridine-4-carboxamide (DACA), d(CGTACG) sub 2 and N-[2-(dimethylamino)ethyl] 9-aminoacridine-4-carboxamide (9- aminoDACA) and some of their derivatives. An attempt was made to solve the structure of the DACA derivative N-[2-(dimethylamino)butyl]-acridine-4-carboxamide (DACA4) by molecular replacement, using the crystal structure of the daunomycin d(CGTACG) sub 2 complex as a search model. Attempts were made to position the molecule in the unit cell based on an SIR map, knowledge of the symmetry and unit cell dimensions. The structure of the 9-amino-5-bromo DACA - d(CGT(5-BrU)CG) su...

  5. Optimization of the Production of Extracellular Polysaccharide from the Shiitake Medicinal Mushroom Lentinus edodes (Agaricomycetes) Using Mutation and a Genetic Algorithm-Coupled Artificial Neural Network (GA-ANN).

    Science.gov (United States)

    Adeeyo, Adeyemi Ojutalayo; Lateef, Agbaje; Gueguim-Kana, Evariste Bosco

    2016-01-01

    Exopolysaccharide (EPS) production by a strain of Lentinus edodes was studied via the effects of treatments with ultraviolet (UV) irradiation and acridine orange. Furthermore, optimization of EPS production was studied using a genetic algorithm coupled with an artificial neural network in submerged fermentation. Exposure to irradiation and acridine orange resulted in improved EPS production (2.783 and 5.548 g/L, respectively) when compared with the wild strain (1.044 g/L), whereas optimization led to improved productivity (23.21 g/L). The EPS produced by various strains also demonstrated good DPPH scavenging activities of 45.40-88.90%, and also inhibited the growth of Escherichia coli and Klebsiella pneumoniae. This study shows that multistep optimization schemes involving physical-chemical mutation and media optimization can be an attractive strategy for improving the yield of bioactives from medicinal mushrooms. To the best of our knowledge, this report presents the first reference of a multistep approach to optimizing EPS production in L. edodes. PMID:27649726

  6. Cytotoxic evaluation of different fractions of Salvia chorassanica Bunge on MCF-7 and DU 145 cell lines

    Directory of Open Access Journals (Sweden)

    Alireza Golshan

    2016-01-01

    Full Text Available Because of antimicrobial, antioxidant, and anticancer potential, Salvia chorassanica Bunge (Lamiaceae has been considered as a popular herb in Iranian traditional medicine. Previous studies have shown remarkable cytotoxic properties of the methanol, n-hexane and dichloromethane extract of S. chorassanica on human cervical cancer cells. To seek the therapeutic potentials of S. chorassanica, this study was undertaken to evaluate the cytotoxic activities of various extracts of this plant on human breast MCF-7 and prostate cancer DU 145 cells. The DU 145 cells were exposed to different concentrations of plant extracts (1-200 μg/ml. Cytotoxic activities were examined using alamarBlue ® assay and apoptosis was assessed by acridine orange/propodium iodide double staining and evaluation of DNA fragmentation by flow cytometry. Our findings indicated that n-hexane and dichloromethane extracts had more cytotoxic activities against DU 145 and MCF-7 cell lines compared with other extracts (P<0.05. The acridine orange/propodium iodide staining showed apoptogenic properties of n-hexane and dichloromethane extracts which was consequently confirmed by flow cytometric histogram that exhibited an increase in sub-G1 peak in treated cells as compared with untreated cancer cell lines. Taken together, these observations demonstrated cytotoxic effects of S. chorassanica extracts on MCF-7 and DU 145 cell lines which is most likely exerted via apoptosis cell death. Therefore, further investigations on S. chorassanica extracts as potential chemotherapeutic agents are warranted.

  7. Characterization of osteoclasts from patients harboring a G215R mutation in ClC-7 causing autosomal dominant osteopetrosis type II

    DEFF Research Database (Denmark)

    Henriksen, Kim; Gram, Jeppe; Schaller, Sophie;

    2004-01-01

    from ADOII patients and healthy age- and sex-matched controls, were used to evaluate osteoclastogenesis, cell fusion, acidification, and resorptive activity. ADOII osteoclasts in vivo have increased number and size. However, in vitro we observed no significant changes in the osteoclast formation rate......Autosomal dominant osteopetrosis II (ADOII) is a relatively benign disorder caused by a missense mutation in the ClCN7 gene. In this study, we characterize the osteoclasts from patients with ADOII, caused by a G215R mutation, and investigate the effect on osteoclast function in vitro. Osteoclasts......, the morphology, and the expression of markers, such as cathepsin K and tartrate-resistant acid phosphatase. When mature ADOII osteoclasts were investigated on mineralized bone, they degraded the bone material, however only to 10 to 20% of the level in controls. We show by acridine orange, that the reduced...

  8. Ion transporters involved in acidification of the resorption lacuna in osteoclasts

    DEFF Research Database (Denmark)

    Henriksen, Kim; Sørensen, Mette G; Jensen, Vicki K;

    2008-01-01

    Osteoclasts possess a large amount of ion transporters, which participate in bone resorption; of these, the vacuolar-adenosine trisphosphatase (V-ATPase) and the chloride-proton antiporter ClC-7 acidify the resorption lacuna. However, whether other ion transporters participate in this process...... is currently not well understood. We used a battery of ion channel inhibitors, human osteoclasts, and their subcellular compartments to perform an unbiased analysis of the importance of the different ion transporters for acidification of the resorption lacuna in osteoclasts. CD14(+) monocytes from human...... peripheral blood were isolated, and mature osteoclasts were generated using RANKL and M-CSF. The human osteoclasts were (1) used for acridine orange assays for evaluation of lysosomal acidification, (2) used for bone resorption assays, (3) used for generation of osteoclasts membranes for acid influx...

  9. [Substrate-inhibitory analysis of monoamine oxidase from hepatopancreas of the octopus Bathypolypus arcticus].

    Science.gov (United States)

    Basova, I N; Iagodina, O V

    2012-01-01

    Study of the substrate-inhibitory specificity of mitochondrial monoamine oxidase (MAO) of hepatopancreas of the octopus Bathypolypus arcticus revealed distinctive peculiarities of catalytic properties of this enzyme. The studied enzyme, on one hand, like the classic MAO of homoiothermal animals, is able to deaminate tyramine, serotonin, benzylamine, tryptamine, beta-phenylethylamine, while, on the other hand, deaminates histamine and does not deaminate putrescine--classic substrates of diamine oxidase (DAO). Results of the substrate-inhibitory analysis with use of chlorgiline and deprenyl are indirect proofs of the existence in the octopus hepatopancreas of one molecular MAO form. Semicarbazide and pyronine G turned out to be weak irreversible inhibitors, four derivatives of acridine--irreversible inhibitors of the intermediate effectiveness with respect to the octopus hepatopancreas MAO; specificity of action of inhibitors at deamination of different substrates was equal.

  10. Experimental and theoretical study of hydrodynamic cell lysing of cancer cells in a high-throughput Circular Multi-Channel Microfiltration device

    KAUST Repository

    Ma, W.

    2013-04-01

    Microfiltration is an important microfluidic technique suitable for enrichment and isolation of cells. However, cell lysing could occur due to hydrodynamic damage that may be detrimental for medical diagnostics. Therefore, we conducted a systematic study of hydrodynamic cell lysing in a high-throughput Circular Multi-Channel Microfiltration (CMCM) device integrated with a polycarbonate membrane. HeLa cells (cervical cancer cells) were driven into the CMCM at different flow rates. The viability of the cells in the CMCM was examined by fluorescence microscopy using Acridine Orange (AO)/Ethidium Bromide (EB) as a marker for viable/dead cells. A simple analytical cell viability model was derived and a 3D numerical model was constructed to examine the correlation of between cell lysing and applied shear stress under varying flow rate and Reynolds number. The measured cell viability as a function of the shear stress was consistent with theoretical and numerical predictions when accounting for cell size distribution. © 2013 IEEE.

  11. Spectroscopic studies on the interaction of sodium benzoate, a food preservative, with calf thymus DNA.

    Science.gov (United States)

    Zhang, Guowen; Ma, Yadi

    2013-11-01

    The interaction between sodium benzoate (SB) and calf thymus DNA in simulated physiological buffer (pH 7.4) using acridine orange (AO) dye as a fluorescence probe, was investigated by UV-Vis absorption, fluorescence and circular dichroism (CD) spectroscopy along with DNA melting studies and viscosity measurements. An expanded UV-Vis spectral data matrix was resolved by multivariate curve resolution-alternating least squares (MCR-ALS) approach. The equilibrium concentration profiles and the pure spectra for SB, DNA and DNA-SB complex from the high overlapping composite response were simultaneously obtained. The results indicated that SB could bind to DNA, and hydrophobic interactions and hydrogen bonds played a vital role in the binding process. Moreover, SB was able to quench the fluorescence of DNA-AO complex through a static procedure. The quenching observed was indicative of an intercalative mode of interaction between SB and DNA, which was supported by melting studies, viscosity measurements and CD analysis.

  12. Crosslinking of cellular DNA by nitracrine and furocoumarin derivatives.

    Science.gov (United States)

    Studzian, K; Tołwińska-Stańczyk, Z; Wilmańska, D; Palumbo, M; Gniazdowski, M

    1999-01-01

    The anticancer drug, nitracrine, a 1-nitro-9-aminoalkyl derivative of acridine exhibits potent cytotoxic effects which are due to its metabolic activation, followed by covalent binding to macromolecules--DNA being the target for the drug. The renaturable fraction of DNA from L-1210 cells pretreated with nitracrine is assayed by means of ethidium bromide fluorescence assay and chromatography on hydroxyapatite column. The effect of the drug was compared with furocoumarins of different DNA crosslinking potencies. The existence of crosslinks in DNA upon incubation of cells with nitracrine (1-4 microM) have been confirmed with two different methods under the conditions where 8-methoxypsoralen, a classic crosslinking agent induced the renaturation. The DNA preparation isolated from the drug pretreated cells exhibited decreased transcriptional template activity with E. coli DNA-dependent RNA polymerase. PMID:10355534

  13. Nitracrine and its congeners--an overview.

    Science.gov (United States)

    Gniazdowski, M; Szmigiero, L

    1995-05-01

    1. An anticancer drug, nitracrine 1-nitro-9(3'3'-dimethylaminopropylamino)acridine (Ledakrin, C-283) exhibits potent cytostatic effects which can be ascribed to interactions of the drug with DNA. 2. The reduction of the nitro group of nitracrine is one of the activation steps leading to the drug covalent binding to DNA and proteins both in subcellular systems and in the cell. 3. DNA-drug non-covalent interactions and covalent complexes are examined in several model systems and compared with the properties of a number of derivatives with programmed structural changes. 4. DNA-protein crosslinks and interstrand crosslinks are detected in the cells following exposition to the drug. 5. The drug exhibits selective toxicity and radiosensitization effects to hypoxic mammalian cells. PMID:7789719

  14. Inhibitory Effect of Melatonin on the Growth of H22 Hepatocarcinoma Cells by Inducing Apoptosis

    Institute of Scientific and Technical Information of China (English)

    泰莉; 王西明; 段秋红; 陈蓓蓓; 何善述

    2004-01-01

    Summary: Whether melatonin not only inhibits the growth of H22 hepatocarcinoma cells but also induces apoptosis in vitro was assessed. The anti-proliferative effects of melatonin on tumor cells was observed by MTT assay and tumor cells growth curve assay. And the apoptosis of the cells was studied by acridine orange fluorescence assay and flow cytometry. The cell cycle of the tumor cells was also observed by flow cytometry. It was found that melatonin could significantly inhibit the growth of H22 hepatocarcinoma cells. Incubated with melatonin, chromatin condensation of the tumor cells was observed by fluorescence microscopy. Compared with control, the percentage of apoptotic cells was increased, and the proportion of G0/S increased but that of G2/M decreased. It was suggested that melatonin could directly inhibit the growth of H22 hepatocarcinoma cells by inducing apoptosis and extending the length of cell cycle of the tumor cells.

  15. Alpha-particle-induced bystander effects between zebrafish embryos in vivo

    Energy Technology Data Exchange (ETDEWEB)

    Yum, E.H.W.; Choi, V.W.Y.; Nikezic, D. [Department of Physics and Materials Science, City University of Hong Kong, Tat Chee Avenue, Kowloon Tong (Hong Kong); Li, V.W.T.; Cheng, S.H. [Department of Biology and Chemistry, City University of Hong Kong, Tat Chee Avenue, Kowloon Tong (Hong Kong); Yu, K.N., E-mail: peter.yu@cityu.edu.h [Department of Physics and Materials Science, City University of Hong Kong, Tat Chee Avenue, Kowloon Tong (Hong Kong)

    2009-10-15

    Dechorionaed embryos of the zebrafish, Danio rerio, at 1.5 h post-fertilization (hpf) were irradiated with alpha particles from an {sup 241}Am source. Thin polyallyldiglycol carbonate (PADC) films with a thickness of 16 mum were used as support substrates for holding the embryos and recorded alpha-particle hit positions, and thus enabled calculation of the dose absorbed by the embryos. The irradiated embryos were subsequently incubated with naive (unirradiated) embryos in such a way that the irradiated and naive embryos were spatially separated but the medium was shared. Acridine orange was used to perform in vital staining to show cell deaths in the naive embryos at 24 hpf. Our results gave evidence in supporting the existence of alpha-particle-induced bystander effects between zebrafish embryos in vivo, and a general positive correlation between the cell death signals in the naive embryos and the alpha-particle dose absorbed by the irradiated embryos.

  16. Energy transfer mechanisms in photobiological reactions. Final report, 1 April 1960--31 March 1979. [Photodynamic processes in selected biomolecules

    Energy Technology Data Exchange (ETDEWEB)

    Spikes, J.D.

    1979-03-31

    This project was concerned primarily with studies of the mechanisms of the sensitized photooxidation of selected biomolecules using a variety of phtosensitizers. Such reactions are often termed photodynamic processes. In particular we have carried out steady-state kinetic studies, flash photolysis and spectral studies, and product formation studies of the sensitized photooxidation of the five susceptible amino acids (cycteine, histidine, methonine, tryptophan, and tyrosine) and their derivatives, as well as purines and pyrimidines. A number of studies were also carried out on the mechanisms of the photodynamic inactivation of enzymes (trypsin, ribonuclease, lysozyme). Mechanism of photosensitization were studied using a variety of sensitizers including flavins, porphyrins, and a number of synthetic dyes (substituted fluoresceins, acridines, thyazines).

  17. Comparison of DNA Fragmentation Assay in Frozen-Thawed Cat Epididymal Sperm.

    Science.gov (United States)

    Kunkitti, P; Sjödahl, A; Bergqvist, A-S; Johannisson, A; Axnér, E

    2016-08-01

    DNA fragmentation of frozen-thawed feline epididymal sperm from corpus and cauda regions was evaluated by three different techniques. The DNA fragmentation index (DFI) was compared between techniques: the sperm chromatin structural assay (SCSA(®) ), acridine orange staining techniques (AOT) and the sperm chromatin dispersion (SCD). There were significant differences in DFI among the techniques (p < 0.05) with no correlations. Only DFI values obtained from SCD revealed a significantly higher DFI in corpus compared with cauda spermatozoa (p < 0.05). The discrepancy between techniques might be due to the sensitivity of each technique, differences in severity of DNA damaged that can be detected. The difference in DFI between epididymal regions from SCD technique might indicate different maturational stages of spermatozoa, with less chromatin condensation of spermatozoa in corpus compared with cauda epididymis. PMID:27321406

  18. Importance of Unattached Bacteria and Bacteria Attached to Sediment in Determining Potentials for Degradation of Xenobiotic Organic Contaminants in an Aerobic Aquifer

    DEFF Research Database (Denmark)

    Nielsen, Per Henning; Albrechtsen, Hans-Jørgen; Christensen, Thomas Højlund;

    1992-01-01

    The bacterial abundance, distribution, and degradation potential (in terms of degradation versus lack of degradation) for four xenobiotic compounds in an aerobic aquifer sediment have been examined in laboratory and field experiments. The xenobiotic compounds studied were benzene, toluene, o......-xylene, and naphthalene (all at concentrations of approximately 120 pg/liter). The aerobic degradation experiments ran for approximately 90 days at 10°C, which corresponded to the groundwater temperature. At the end of the experiment, the major part of the microbial biomass, quantified as acridine orange direct counts...... for studying the degradation potential for xenobiotic organic contaminants should contain sediment to obtain the highest numbers of bacteria as well as the broadest and most stable degradation. When only the fine (silt- and clay-size) particles of the sediment were used, nearly the same advantages were gained...

  19. Characterization of anticancer, DNase and antifungal activity of pumpkin 2S albumin.

    Science.gov (United States)

    Tomar, Prabhat Pratap Singh; Nikhil, Kumar; Singh, Anamika; Selvakumar, Purushotham; Roy, Partha; Sharma, Ashwani Kumar

    2014-06-13

    The plant 2S albumins exhibit a spectrum of biotechnologically exploitable functions. Among them, pumpkin 2S albumin has been shown to possess RNase and cell-free translational inhibitory activities. The present study investigated the anticancer, DNase and antifungal activities of pumpkin 2S albumin. The protein exhibited a strong anticancer activity toward breast cancer (MCF-7), ovarian teratocarcinoma (PA-1), prostate cancer (PC-3 and DU-145) and hepatocellular carcinoma (HepG2) cell lines. Acridine orange staining and DNA fragmentation studies indicated that cytotoxic effect of pumpkin 2S albumin is mediated through induction of apoptosis. Pumpkin 2S albumin showed DNase activity against both supercoiled and linear DNA and exerted antifungal activity against Fusarium oxysporum. Secondary structure analysis by CD showed that protein is highly stable up to 90°C and retains its alpha helical structure. These results demonstrated that pumpkin 2S albumin is a multifunctional protein with host of potential biotechnology applications. PMID:24814706

  20. Agonists that increase [Ca2+]i halt the movement of cytoplasmatic vesicles in MDCK cells

    DEFF Research Database (Denmark)

    Bjælde, Randi Groslier; Árnadóttir, Sigrid Salling; Leipziger, Jens Georg;

    2011-01-01

    loaded with either quinacrine or acridine orange, dyes taken up by acidic vesicles, were observed at 37°C in semiopen perfusion chambers. Time-lapse series were analyzed by Imaris software. Our data revealed vigorous movement of stained vesicles in resting MDCK cells. These movements seem to require...... of vesicles by 40%. Because all these perturbations increase [Ca²⁺]i, we speculated that this increase in [Ca²⁺]i was responsible for the vesicle arrest. Therefore, we tested the effect of the Ca²⁺ ionophore, ionomycin (1 μM), which in the presence of extracellular Ca²⁺ resulted in a considerable...

  1. Determination of antimutagenic properties of apigenin-7-O-rutinoside, a flavonoid isolated from Mentha longifolia (L.) Huds. ssp. longifolia with yeast DEL assay.

    Science.gov (United States)

    Gulluce, Medine; Orhan, Furkan; Adiguzel, Ahmet; Bal, Tugba; Guvenalp, Zuhal; Dermirezer, Lutfiye Omur

    2013-07-01

    Lamiaceae is an important plant family that has been investigated for its medicinal properties due to its large amounts of phenolic acids and flavonoids. Flavonoids have been shown to have antioxidant and antimutagenic activities in different test systems, but their certain mechanisms are still unclear. This study was designed to evaluate the mutagenic and antimutagenic activities of apigenin 7-O-rutinoside, a flavonoid isolated from Mentha longifolia (L.) Huds. ssp. longifolia. The possible antimutagenic potential of apigenin 7-O-rutinoside (A7R) was examined against mutagens ethyl methanesulfonate (EMS) and acridine (AC) in a eukaryotic cell system Saccharomyces cerevisiae RS112. The results showed that A7R has different inhibition rates against EMS and AC-induced mutagenicity. Thus, the properties of A7R are of great pharmacological importance and might be beneficial for reducing the risk of reactive oxygen species-related diseases.

  2. Investigation of cytotoxic activity on human cancer cell lines of arborinine and furanoacridones isolated from Ruta graveolens.

    Science.gov (United States)

    Réthy, Borbála; Zupkó, István; Minorics, Renáta; Hohmann, Judit; Ocsovszki, Imre; Falkay, George

    2007-01-01

    The cytotoxic effects of a series of furanoacridones isolated from Ruta graveolens L. (Rutaceae) and of two further acridone alkaloids (arborinine and evoxanthine) were investigated by means of the MTT assay, using the human cell lines HeLa, MCF7 and A431. Arborinine proved best in inhibiting the proliferation of all three cell lines. The cytotoxic potency of the furacridone alkaloids was a function of their lipid solubility, which was determined by means of PAMPA. The capacity of the most effective furanoacridones to induce apoptosis was demonstrated by flow cytometric cell cycle analysis and by staining with ethidium bromide and acridine orange. This finding was reinforced by determining the apoptosis-regulating factors Bcl-2 and Bax, which were revealed by means of RT-PCR to change dose-dependently. The data presented here indicate that naturally occurring furanoacridones can be regarded as excellent starting structures for the potential development of new anticancer agents.

  3. Laboratory simulation of meteoritic noble gases. I - Sorption of xenon on carbon: Trapping experiments

    Science.gov (United States)

    Wacker, J. F.; Zadnik, M. G.; Anders, E.

    1985-01-01

    The sorption of Xe-127 at 5 x 10 to the -7th atm onto carbon black, pyrolyzed polyvinylidene chloride, and pyrolyzed acridine at 100-1000 C for 5 min-240 h is measured experimentally by gamma spectrometry. The results are presented in tables and graphs and characterized in detail. The tightly bound Xe remaining in the samples after 4000 min pumping at temperatures above 100 C is found to comprise two components: a low-temperature component attributed to physisorption within an atomic-scale labyrinth of micropores, and a high-temperature component due to volume diffusion. The implications for the trapping of noble gases near grain surfaces of amorphous carbon in meteorites are considered.

  4. A NEW MODEL OF BOAR SEMEN EVALUATION AND THE IMPACT OF CRYOGENIC FACTOR ON SPERMATIC CELLS

    Directory of Open Access Journals (Sweden)

    A. V. RUSU

    2013-07-01

    Full Text Available Nowadays, sperm evaluation is mostly used to predict fertility and freezability. The aim of this study is to evaluate the possibility of investigating the effects of the cryogenic agent on boar spermatozoa, by identifying a set of laboratory tests for a rapid and efficient evaluation of semen quality. Usual sperm analysis such as sperm concentration, motility and spermatozoa morphology are not able to show subtle abnormalities, which are having a basic role in the fertilizing ability. Moreover, it seems that other sperm characteristics, involved in the fertilizing ability, can interfere with the freezing-thawing processes, being not evaluated or maybe not known. Morphological (microscopic analysis of stained spermatozoa, functional (motility analysis and hypo-osmotic swelling test and chromatin integrity (Acridine Orange Test and Comet Assay analysis were performed aiming to show the differences in spermatozoon integrity and functionality, caused by the cryogenic factor.

  5. A study of Nigella sativa induced growth inhibition of MCF and HepG2 cell lines: An anti-neoplastic study along with its mechanism of action

    Directory of Open Access Journals (Sweden)

    Y Padmanabha Reddy

    2015-01-01

    Full Text Available Objective: To evaluate the anticancer potential of seeds of Nigella sativa using MCF and HepG2 cell lines along with its mechanism of action. Materials and Methods: (3-(4,5-dimethylthiazol-2-yl-2,5-diphenyltetrazolium bromide assay and acridine orange/ethidium bromide nuclear staining technique were selected to evaluate anticancer potential and mechanism of action of test extract. Results: Aqueous extract of N.sativa at a test dose of 180 mg and 300 mg was identified to be the best as anticancer agent against MCF and HepG2 cell lines among different solvent test extract where doxorubicin and cisplatin were employed as standard references. Discussion: Further study including separation and characterization of active principles in the aqueous extract shall prove beneficial.

  6. [The characteristic of proliferative activity of thymocytes and peripheral blood lymphocytes in the offspring of females with experimental chronic liver diseases of various aetiology].

    Science.gov (United States)

    Briukhin, G V; Fedosov, A A

    2006-01-01

    The aim of the study was a comparative analysis of the proliferative activity of thymocytes and peripheral blood lymphocytes in the offspring of female rats with chronic liver pathology of various genesis. In adult female Wistar rats toxic and autoimmune forms of liver lesions were modeled. The offspring of these experimental animals was studied at different time points of postnatal ontogenesis. Proliferative activity of thymocytes and lymphocytes was estimated by counting the proportion of cells with multiple nucleolar organizing regions (AgNORs) and using the cytofluorometric method with acridine orange. In the offspring of experimental animals, the depression of proliferative activity of thymocytes as well as the increase of the proliferative activity of peripheral blood lymphocytes were found at all the time points studied. This was indicated by a change in a relative number of AgNORs-activated cells and a decrease of nucleic acid content in cortical thymocytes. PMID:17201321

  7. Deoxyamphimedine, a Pyridoacridine Alkaloid, Damages DNA via the Production of Reactive Oxygen Species

    Directory of Open Access Journals (Sweden)

    Chris M. Ireland

    2009-05-01

    Full Text Available Marine pyridoacridines are a class of aromatic chemicals that share an 11H-pyrido[4,3,2-mn]acridine skeleton. Pyridoacridine alkaloids display diverse biological activities including cytotoxicity, fungicidal and bactericidal properties, production of reactive oxygen species (ROS and topoisomerase inhibition. These activities are often dependent on slight modifications to the pyridoacridine skeleton. Here we demonstrate that while structurally similar to neoamphimedine and amphimedine, the biological activity of deoxyamphimedine differs greatly. Deoxyamphimedine damages DNA in vitro independent of topoisomerase enzymes through the generation of reactive oxygen species. Its activity was decreased in low oxygen, with the removal of a reducing agent and in the presence of anti-oxidants. Deoxyamphimedine also showed enhanced toxicity in cells sensitive to single or double strand DNA breaks, consistent with the in vitro activity.

  8. Study of anti mutagenic and mutagenic effect of different chemicals on clinically isolated strains of pseudomonas aeruginosa

    International Nuclear Information System (INIS)

    This project was undertaken to study the effect of twelve different compounds to test their anti mutagenic and mutagenic activity against clinically isolated strains of Pseudomonas aeruginosa. The effect of these compounds was estimated by counting the number of rifampicin resistant colonies growing in a particular time in a compound. The results were interpreted by plotting graphs between 10g N/NO (Rif R Colonies/ ml) and time to estimate the forward mutation rat. The results revealed that acridine, Basic fuchsin, Caffeine, cycloheximide, Ethidium bromide and Histidine probably have an anti mutagenic effect, while Cysteine, folic acid, Ethyl methane, suplphonate, Manganous Chloride and N-nitrosodietylamine acted as mutagen. Ecoli was used as control through out the study. (author)

  9. Induction of apoptosis in human endothelial cells by nanodiamond particles.

    Science.gov (United States)

    Solarska, K; Gajewska, A; Bartosz, G; Mitura, K

    2012-06-01

    Carbon nanoparticles are a promising material which finds application in different fields in industry and medicine. For medical applications, biocompatibility of nanoparticles is of critical importance because a lot of medical implants are coated by carbon coating. Our previous results showed that nanoparticles may induce increased production of ROS by the cells so we decided to checked if nanopowders can induce apoptosis. Apoptosis was quantified by double-staining with acridine orange and ethidium bromide. For comparison, we identified apoptotic cells with annexin V-FITC/propidium iodide. Our data demonstrate that treatment of the cells with diamond nanopowders may induce apoptosis and necrosis and this effect is dependent on the time of treatment and concentration of the nanopowders. The highest level of apoptotic cells was observed after incubation with Ultrananocrystalline Detonation Diamond (UDD) suggesting that the size is the main determinant of nanoparticle cytotoxicity. PMID:22905588

  10. Library of UV-Vis-NIR reflectance spectra of modern organic dyes from historic pattern-card coloured papers.

    Science.gov (United States)

    Montagner, Cristina; Bacci, Mauro; Bracci, Susanna; Freeman, Rachel; Picollo, Marcello

    2011-09-01

    An accurate characterisation of the organic dyes used in artworks, especially those made of paper, is an important factor in designing safe conservation treatments. In the case of synthetic organic dyes used in modern works of art, for example, one frequently encountered difficulty is that some of these dyes are not still commercially available. Recognizing this problem, the authors of this paper present the results of an analysis of UV-Vis-NIR fibre optic reflectance spectra of 82 samples of dyed paper prepared with 41 dyes. The samples come from a historic book, The Dyeing of Paper in the Pulp, which was published by Interessen-Gemeinschaft (I.G.) Farbenindustrie in 1925. The dyes used in the paper pulp belong to the azo compounds, acridine, anthraquinone, azine, diphenylmethane, indigoid, methine, nitro, quinoline, thiazine, triphenylmethane, sulphur and xanthene classes.

  11. RutheniumII Complexes bearing Fused Polycyclic Ligands: From Fundamental Aspects to Potential Applications

    Directory of Open Access Journals (Sweden)

    Ludovic Troian-Gautier

    2014-04-01

    Full Text Available In this review, we first discuss the photophysics reported in the literature for mononuclear ruthenium complexes bearing ligands with extended aromaticity such as dipyrido[3,2-a:2',3'-c]phenazine (DPPZ, tetrapyrido[3,2-a:2',3'-c:3'',2''-h:2''',3'''-j]-phenazine (TPPHZ,  tetrapyrido[3,2-a:2',3'-c:3'',2''-h:2''',3'''-j]acridine (TPAC, 1,10-phenanthrolino[5,6-b]1,4,5,8,9,12-hexaazatriphenylene (PHEHAT 9,11,20,22-tetraaza- tetrapyrido[3,2-a:2',3'-c:3'',2''-l:2''',3'''-n]pentacene (TATPP, etc. Photophysical properties of binuclear and polynuclear complexes based on these extended ligands are then reported. We finally develop the use of binuclear complexes with extended π-systems for applications such as photocatalysis.

  12. Carbamazepine degradation by photolysis and titanium dioxide photocatalysis.

    Science.gov (United States)

    Im, Jong-Kwon; Son, Hyun-Seok; Kang, Young-Min; Zoh, Kyung-Duk

    2012-07-01

    We investigated the degradation of carbamazepine by photolysis/ultraviolet (UV)-C only and titanium dioxide photocatalysis. The degradation of carbamazepine by UV-only and titanium-dioxide-only (adsorption) reactions were inefficient, however, complete degradation of carbamazepine was observed by titanium dioxide photocatalysis within 30 min. The rate of degradation increased as initial carbamazepine concentration decreased, and the removal kinetics fit well with the Langmuir-Hinshelwood model. The addition of methanol, a radical scavenger, decreased carbamazepine removal, suggesting that the hydroxide radical played an important role during carbamazepine degradation. The addition of oxygen during titanium dioxide photocatalysis accelerated hydroxide radical production, thus improving mineralization activity. The photocatalytic degradation was more efficient at a higher pH, whereas the removal of carbamazepine and acridine (a major intermediate) were more efficient under aerobic conditions. The mineralization of carbamazepine during photocatalysis produced various ionic by-products such as ammonium and nitrate by way of nitrogen dioxide.

  13. Role of cloned carotenoid genes expressed in Escherichia coli in protecting against inactivation by near-UV light and specific phototoxic molecules

    International Nuclear Information System (INIS)

    Genes controlling carotenoid synthesis were cloned from Erwinia herbicola and expressed in an Escherichia coli strain. Carotenoids protect against high fluences of near-UV (NUV; 320 to 400 nm) but not against far-UV (200-300 nm). Protection of E. coli cells was not observed following treatment with either psoralen or 8-methoxypsoralen plus NUV. However, significant protection of cells producing carotenoids was observed with three photosensitizing molecules activated by NUV (alpha-terthienyl, harmine, and phenylheptatriyne) which are thought to have the membrane as an important lethal target. Protection of carotenoid-producing cells against inactivation was not observed with acridine orange plus visible light but was seen with toluidine blue O plus visible light

  14. Role of cloned carotenoid genes expressed in Escherichia coli in protecting against inactivation by near-UV light and specific phototoxic molecules

    Energy Technology Data Exchange (ETDEWEB)

    Tuveson, R.W.; Larson, R.A.; Kagan, J.

    1988-10-01

    Genes controlling carotenoid synthesis were cloned from Erwinia herbicola and expressed in an Escherichia coli strain. Carotenoids protect against high fluences of near-UV (NUV; 320 to 400 nm) but not against far-UV (200-300 nm). Protection of E. coli cells was not observed following treatment with either psoralen or 8-methoxypsoralen plus NUV. However, significant protection of cells producing carotenoids was observed with three photosensitizing molecules activated by NUV (alpha-terthienyl, harmine, and phenylheptatriyne) which are thought to have the membrane as an important lethal target. Protection of carotenoid-producing cells against inactivation was not observed with acridine orange plus visible light but was seen with toluidine blue O plus visible light.

  15. Surface modification of low cost carbons for their application in the environmental protection

    Energy Technology Data Exchange (ETDEWEB)

    Arenillas, A. [Instituto Nacional del Carbon, CSIC, Apartado 73, 33080 Oviedo (Spain)]. E-mail: aapuente@incar.csis.es; Rubiera, F. [Instituto Nacional del Carbon, CSIC, Apartado 73, 33080 Oviedo (Spain); Parra, J.B. [Instituto Nacional del Carbon, CSIC, Apartado 73, 33080 Oviedo (Spain); Ania, C.O. [Instituto Nacional del Carbon, CSIC, Apartado 73, 33080 Oviedo (Spain); Pis, J.J. [Instituto Nacional del Carbon, CSIC, Apartado 73, 33080 Oviedo (Spain)

    2005-10-31

    In this work, the CO{sub 2} capture capacity of a series of activated carbons derived from recycled polyethylene terephtalate (PET) was tested, facing two problems at the same time: minimising plastic waste and developing an adsorbent for CO{sub 2} capture. The PET raw material, obtained from post-consumer soft-drink bottles, was chemically activated with KOH. In addition, a series of nitrogen-enriched activated carbons was obtained by mixing the raw material with different nitrogen compounds (i.e., acridine, carbazole and urea). The influence of temperature on the CO{sub 2} capture capacity of the adsorbents was evaluated in a thermogravimetric system. The CO{sub 2} uptake was also related to the chemical and textural characteristics of the samples.

  16. Surface modification of low cost carbons for their application in the environmental protection

    Science.gov (United States)

    Arenillas, A.; Rubiera, F.; Parra, J. B.; Ania, C. O.; Pis, J. J.

    2005-10-01

    In this work, the CO 2 capture capacity of a series of activated carbons derived from recycled polyethylene terephtalate (PET) was tested, facing two problems at the same time: minimising plastic waste and developing an adsorbent for CO 2 capture. The PET raw material, obtained from post-consumer soft-drink bottles, was chemically activated with KOH. In addition, a series of nitrogen-enriched activated carbons was obtained by mixing the raw material with different nitrogen compounds (i.e., acridine, carbazole and urea). The influence of temperature on the CO 2 capture capacity of the adsorbents was evaluated in a thermogravimetric system. The CO 2 uptake was also related to the chemical and textural characteristics of the samples.

  17. Biodegradation of bisphenol A and decolorization of synthetic dyes by laccase from white-rot fungus, Trametes polyzona.

    Science.gov (United States)

    Chairin, Thanunchanok; Nitheranont, Thitinard; Watanabe, Akira; Asada, Yasuhiko; Khanongnuch, Chartchai; Lumyong, Saisamorn

    2013-01-01

    Purified laccase from Trametes polyzona WR710-1 was used as biocatalyst for bisphenol A biodegradation and decolorization of synthetic dyes. Degradation of bisphenol A by laccase with or without redox mediator, 1-hydroxybenzotriazole (HBT) was studied. The quantitative analysis by HPLC showed that bisphenol A rapidly oxidized by laccase with HBT. Bisphenol A was completely removed within 3 h and 4-isopropenylphenol was found as the oxidative degradation product from bisphenol A when identified by GC-MS. All synthetic dyes used in this experiment, Bromophenol Blue, Remazol Brilliant Blue R, Methyl Orange, Relative Black 5, Congo Red, and Acridine Orange were decolorized by Trametes laccase and the percentage of decolorization increased when 2 mM HBT was added in the reaction mixture. This is the first report showing that laccase from T. polyzona is an affective enzyme having high potential for environmental detoxification, bisphenol A degradation and synthetic dye decolorization.

  18. Fabrication and non-covalent modification of highly oriented thin films of a zeolite-like metal-organic framework (ZMOF) with rho topology

    KAUST Repository

    Shekhah, Osama

    2015-01-01

    Here we report the fabrication of the first thin film of a zeolite-like metal-organic framework (ZMOF) with rho topology (rho-ZMOF-1, ([In48(HImDC)96]48-)n) in a highly oriented fashion on a gold-functionalized substrate. The oriented rho-ZMOF-1 film was functionalized by non-covalent modification via post-synthetic exchange of different probe molecules, such as acridine yellow, methylene blue, and Nile red. In addition, encapsulation of a porphyrin moiety was achieved via in situ synthesis and construction of the rho-ZMOF. Adsorption kinetics of volatile organic compounds on rho-ZMOF-1 thin films was also investigated. This study suggests that rho-ZMOF-1 thin films can be regarded as a promising platform for various applications such as sensing and catalysis. This journal is

  19. Mercury decreases culturability of Pseudomonas frederiksbergensis JAJ 28 in soil microcosms

    DEFF Research Database (Denmark)

    Johnsen, Kaare; Ekelund, Flemming; Binnerup, Svend J;

    2003-01-01

    Mercury is a biologically potent heavy metal, which has been found to change the diversity of culturable bacteria. Therefore, we investigated whether Hg kills bacteria in soil or reduces culturability. Soil microcosms were inoculated with Pseudomonas frederiksbergensis JAJ 28 and were sampled...... regularly during 28 days. The total number of acridine orange-stained cells was relatively constant, and Hg reduced the number on only one sampling day. However, the fraction of culturable cells on 1/10 tryptic soy agar was lowered on days 6, 13, and 21. The number of microcolony forming units, which...... represents viable cells, was also affected by Hg, but this effect was delayed compared with the effects on CFUs. The amount of headspace CO2 per cell was overall increased by Hg, another indication of the toxic effects of Hg on the bacterial cells. Our results thus emphasize the need to take culturability...

  20. Synthesis and Study of Second-order Nonlinear Optical Properties of 3-Substituted-6- (substituted-phenylazo) coumarins

    Institute of Scientific and Technical Information of China (English)

    SONG Hua-Can; WEN Huan; LIANG Dong; SUN Yi-Feng

    2003-01-01

    @@ It has attracted a lot of attentions to synthesize and investigate the behaviors of organic second-order nonlinear optical (NLO) materials. [1,2] We have ever reported that acridine derivatives ,[3] 4-substituted-benzylideneoxazol-5(4H)-one[4] and 4,4′-di-styryl-biphenyl derivatives[5] possess good second-order NLO properties. Coumarin derivatives are good organic optical materials and azobenzene derivatives possess a higher second-order nonlinear polarization values, however, there are few reports about the study on the synthetic method, optical behavior, especially,second-order NLO properties of 3-substitued-6-(substituted-phenylazo) coumarin derivatives. Therefore, a series of the following compounds were prepared in order to investigate their NLO behavior.

  1. A fibroblast-associated antigen: Characterization in fibroblasts and immunoreactivity in smooth muscle differentiated stromal cells

    DEFF Research Database (Denmark)

    Rønnov-Jessen, Lone; Celis, Julio E.; van Deurs, Bo;

    1992-01-01

    from vascular smooth muscle cells. The antigen was detected on the cell surface and in cathepsin D-positive and acridine orange-accumulating vesicular compartments of fibroblasts. Ultrastructurally, the antigen was revealed in coated pits and in endosomal and lysosomal structures. 1B10 recognized three...... major brands migrating at apparent Mr of 38,000, 45,000, and 80,000, in addition to many minor bands between Mr 45,000 and 97,000, including Mr 52,000. The Mr 45,000 and 38,000 were associated with the cell membrane and Mr 52,000 as well as Mr 38,000 were associated with the lysosomes. The 1B10...

  2. Effect of the Vacuolation of Helicobacter Pylori

    Institute of Scientific and Technical Information of China (English)

    2001-01-01

    Cytotoxic test in vitro combined with cytochemical stain, fluorescent stain, transmission electronmicrograph was used to study the vacuolated effect by helicobacter pylori (H.pylori) (Toxin+) and its pathological mechanism. 78.26 % patients with peptic ulcer associated with H.pylori was infected with H.pylori (Toxin+), while 42.86 % patients with gastritis was infected with H.pylori (Toxin+). It was positive in vacuole with acridine orange and acid phosphatase stain. Transmission electronmicrograph of vacuole revealed the presence of abounding membrane. There was a closed relationship between infection with H.pylori (Toxin+) and peptic ulcer disease. The vacuole induced by H.pylori (Toxin+) was autophagosome, which was pathological phenomenon induced by toxin.

  3. Biphasic and triphasic dose responses in zebrafish embryos to low-dose 150 kV X-rays with different levels of hardness.

    Science.gov (United States)

    Kong, Eva Yi; Cheng, Shuk Han; Yu, Kwan Ngok

    2016-07-01

    The in vivo low-dose responses of zebrafish (Danio rerio) embryos to 150 kV X-rays with different levels of hardness were examined through the number of apoptotic events revealed at 24 h post fertilization by vital dye acridine orange staining. Our results suggested that a triphasic dose response was likely a common phenomenon in living organisms irradiated by X-rays, which comprised an ultra-low-dose inhibition, low-dose stimulation and high-dose inhibition. Our results also suggested that the hormetic zone (or the stimulation zone) was shifted towards lower doses with application of filters. The non-detection of a triphasic dose response in previous experiments could likely be attributed to the use of hard X-rays, which shifted the hormetic zone into an unmonitored ultra-low-dose region. In such cases where the subhormetic zone was missed, a biphasic dose response would be reported instead.

  4. Nanophotonics and DNA: New approaches

    CERN Document Server

    Bregadze, Vasil G; Giorgadze, Tamar G

    2014-01-01

    The aim of the present work is spectroscopic and thermodynamic study of DNA catalytic properties in the following processes: a) redox; b) formation of inter-strand cross-links; c) performing of photo-dynamic effects; d) nanoscale resonance radiationless electron excitation energy transfer. The most attention is paid to the latter, as truly nanoscale method in its origin. The nanoscale method of laser induced fluorescence resonance energy transfer (FRET) to donor (acridine orange) - acceptor (ethidium bromide) intercalator pair for quantitative and qualitative study of stability quality DNA double helix in solution in real time is offered. FRET method allows to estimate the concentration of double helix areas with high quality stability applicable for intercalation in DNA after it was subjected to stress effect. It gives the opportunity to compare various types of DNAs with 1) different origin; 2) various damage degrees; 3) being in various functional state. Alternative model and mechanisms of photodynamic eff...

  5. A NEW MODEL OF BOAR SEMEN EVALUATION AND THE IMPACT OF CRYOGENIC FACTOR ON SPERMATIC CELLS

    Directory of Open Access Journals (Sweden)

    M. ZĂHAN

    2009-05-01

    Full Text Available Nowadays, sperm evaluation is mostly used to predict fertility and freezability. Theaim of this study is to evaluate the possibility of investigating the effects of thecryogenic agent on boar spermatozoa, by identifying a set of laboratory tests for arapid and efficient evaluation of semen quality. Usual sperm analysis such as spermconcentration, motility and spermatozoa morphology are not able to show subtleabnormalities, which are having a basic role in the fertilizing ability. Moreover, itseems that other sperm characteristics, involved in the fertilizing ability, can interferewith the freezing-thawing processes, being not evaluated or maybe not known.Morphological (microscopic analysis of stained spermatozoa, functional (motilityanalysis and hypo-osmotic swelling test and chromatin integrity (Acridine OrangeTest and Comet Assay analysis were performed aiming to show the differences inspermatozoon integrity and functionality, caused by the cryogenic factor

  6. Microbial biomass and activity in subsurface sediments from Vejen, Denmark

    DEFF Research Database (Denmark)

    Albrechtsen, Hans-Jørgen; Winding, Anne

    1992-01-01

    of bacteria varied from 0.5 to 1,203 x 103 colony forming units/g dry weight (gdw); total numbers of bacteria acridine orange direct counts (AODC) varied from 1.7 to 147 × 107 cells/gdw; growth rates (incorporation of [3H]-thymidine) varied from 1.4 to 60.7 × 104 cells/(gdw · day); and rate constants...... a single abiotic parameter that could explain the variation of size and activity of the microbial population. The microbial data obtained in these geologically young sediments were compared to literature data from older sediments, and this comparison showed that age and type of geological formation might...... be important for the size and activity of the microbial populations....

  7.  Distribution and composition of microbial populations in landfill leachate contaminated aquifer (Grindsted, Denmark)

    DEFF Research Database (Denmark)

    Ludvigsen, L; Albrechtsen, HJ; Ringelberg, DB;

    1999-01-01

    To investigate whether landfill leachates affected the microbial biomass and/or community composition of the extant microbiota, 37 samples were collected along a 305-m transect of a shallow landfill-leachate polluted aquifer. The samples were analyzed for total numbers of bacteria by use...... in PLFA profiles indicated that a shift in microbial community composition occurred with increasing horizontal distance from the landfill. The types and patterns of lipid biomarkers suggested that increased proportions of sulfate- and iron-reducing bacteria as well as certain microeukaryotes existed...... of the acridine orange direct count method (AODC). Numbers of dominant, specific groups of bacteria and total numbers of protozoa were measured by use of the most probable number method (MPN). Viable biomass estimates were obtained from measures of ATP and ester-linked phospholipid fatty acid (PLFA...

  8. Drug-DNA intercalation: from discovery to the molecular mechanism.

    Science.gov (United States)

    Mukherjee, Arnab; Sasikala, Wilbee D

    2013-01-01

    The ability of small molecules to perturb the natural structure and dynamics of nucleic acids is intriguing and has potential applications in cancer therapeutics. Intercalation is a special binding mode where the planar aromatic moiety of a small molecule is inserted between a pair of base pairs, causing structural changes in the DNA and leading to its functional arrest. Enormous progress has been made to understand the nature of the intercalation process since its idealistic conception five decades ago. However, the biological functions were detected even earlier. In this review, we focus mainly on the acridine and anthracycline types of drugs and provide a brief overview of the development in the field through various experimental methods that led to our present understanding of the subject. Subsequently, we discuss the molecular mechanism of the intercalation process, free-energy landscapes, and kinetics that was revealed recently through detailed and rigorous computational studies.

  9. The ultrastructure of shelled and unshelled cashew nuts.

    Science.gov (United States)

    Muniz, Celli R; Freire, Francisco C O; Soares, Arlete Aparecida; Cooke, Peter H; Guedes, Maria I F

    2013-01-01

    Cashew nuts have many attributes, including sensory, nutritional and health appeal, which contribute to their worldwide acceptance. We demonstrate details of the microstructure of shelled and unshelled cashew kernels with regard to pericarp and cotyledon organization. This study also provides evidence of the colonization of these kernels by filamentous fungi. Nuts were examined by scanning electron and confocal scanning laser microscopy. Staining with acridine orange was performed. A tight lignified palisade layer adjacent to the exocarp surface explains the hardness of the shell's pericarp. The mesocarp contains large secretory cavities that confer a spongy property to this tissue. Papillose cells, which are responsible for secreting CNSL (cashew nutshell liquid), were observed to cover the inner wall of these cavities. Lipid components are readily released from the parenchyma and appear as oil droplets. The outer surface of the shelled samples exhibited a dense Aspergillus infestation. PMID:24045033

  10. Assessment of human natural killer and lymphokine-activated killer cell cytotoxicity against Toxoplasma gondii trophozoites and brain cysts

    Energy Technology Data Exchange (ETDEWEB)

    Dannemann, B.R.; Morris, V.A.; Araujo, F.G.; Remington, J.S. (Palo Alto Medical Foundation, CA (USA))

    1989-10-15

    Because previous work has suggested that NK cells may be important in host resistance against the intracellular parasite Toxoplasma gondii we examined whether human NK cells and lymphokine-activated killer (LAK) cells have activity against trophozoites and cysts of this organism in vitro. A method to radiolabel Toxoplasma trophozoites with 51Cr was developed and direct cytotoxic activity was determined by using modifications of the standard 51Cr release assay. Viability of 51Cr-labeled trophozoites assessed by both methylene blue staining and trypan blue exclusion was greater than 90%. Significantly more 51Cr was released by anti-Toxoplasma antibody and C than by antibody in the absence of C. Incubation of trophozoites with freshly isolated human NK cells or NK cells activated with either rIL-2 or rIFN-alpha did not result in significant release of 51Cr (specific lysis was 0 to 2.3%). In contrast, the average specific lysis of radiolabeled trophozoites by LAK cells was significant. In a series of separate experiments, preincubation of radiolabeled trophozoites with heat-inactivated normal or Toxoplasma antibody-positive human serum increased the cytotoxicity of LAK cells from a mean specific lysis of 15% +/- 4.5 to 39% +/- 8.5, respectively, as assessed by 51Cr release. Because previous work has shown that radioisotope release from parasites may be nonspecific, separate experiments were performed to determine the cytotoxicity of LAK cells against antibody-coated trophozoites by using ethidium bromide-acridine orange staining to assess effector cell damage. LAK cells had a mean specific lysis of 51% against antibody-coated trophozoites by ethidium bromide-acridine orange staining. Preincubation with heat-inactivated Toxoplasma-antibody positive human serum did not increase activity of rIL-2-activated NK cells against 51CR-labeled trophozoites.

  11. KAJIAN BIOAKTIF DAN ZAT GIZI PROPOLIS INDONESIA DAN BRASIL

    Directory of Open Access Journals (Sweden)

    Eliza Halim

    2012-11-01

    Full Text Available ABSTRACTIndonesia has a potency to produce its own propolis, however the propolis market in Indonesia is dominated by imported product, such as from Brazil. Currently, still there is no reasearch which evaluate bioactive compound and nutrient content of Indonesian Propolis (IP compare with Brazilian Propolis (BP. The objectivesof this study were to analyze bioactive compounds and nutrient contents of IP compared to BP. Bioactive compounds and nutrients content were analyzed by gas chromatography–mass spectrophotometry. The resultsshowed both IP and BP contain fenol, α-amyrin, cylolanost, and pyrimidines. Bioactive compounds which specifically found in IP were eudesmane compound, ethyl acridine, lupeol, friedooleanan; while β amyrin and cinnamic acid compound only found in BP. The nutrient contents of IP were higher than BP except for vitamin A. In conclusion, IP might have potential health benefit, similar to BP.Key words: propolis, bioactive, nutrientABSTRAKIndonesia mempunyai potensi untuk menghasilkan propolis tetapi pemasaran propolis di Indonesia didominasi oleh propolis impor seperti propolis yang berasal Brasil. Sampai saat ini belum ada penelitian yang mengungkapkandungan bioaktif dan zat gizi propolis Indonesia (PI dibandingkan dengan propolis Brasil (PB. Oleh karena itu tujuan penelitian ini adalah untuk melakukan kajian kandungan bioaktif dan zat gizi (vitamin dan mineral PI dibandingkan dengan PB. Komponen bioaktif dan kandungn gizi dianalisis dengan metode gas chromatography–mass spectrometry. Hasil analisis menyatakan bahwa baik PI maupun PB mengandung senyawa fenol, α-amyrin, cylolanost, dan pirimidin. Komponen bioaktif unik yang ditemukan di dalam PI adalah senyawaeudesmane, ethyl acridine, lupeol dan friedooleanan; sedangkan β-amyrin dan senyawa asam sinamat hanya ditemukan di dalam PB. Kandungan zat gizi PI lebih tinggi dari PB kecuali kandungan vitamin A. Hal ini menunjukkan bahwa PI kemungkinan mempunyai khasiat untuk

  12. Molecular and biochemical alterations in tubular epithelial cells of patients with isolated methylmalonic aciduria.

    Science.gov (United States)

    Ruppert, T; Schumann, A; Gröne, H J; Okun, J G; Kölker, S; Morath, M A; Sauer, S W

    2015-12-15

    Methylmalonic acidurias (MMAurias) are a group of inherited disorders in the catabolism of branched-chain amino acids, odd-chain fatty acids and cholesterol caused by complete or partial deficiency of methylmalonyl-CoA mutase (mut(0) and mut(-) subtype respectively) and by defects in the metabolism of its cofactor 5'-deoxyadenosylcobalamin (cblA, cblB or cblD variant 2 type). A long-term complication found in patients with mut(0) and cblB variant is chronic tubulointerstitial nephritis. The underlying pathomechanism has remained unknown. We established an in vitro model of tubular epithelial cells from patient urine (hTEC; 9 controls, 5 mut(0), 1 cblB). In all human tubular epithelial cell (hTEC) lines we found specific tubular markers (AQP1, UMOD, AQP2). Patient cells showed disturbance of energy metabolism in glycolysis, mitochondrial respiratory chain and Krebs cycle in concert with increased reactive oxygen species (ROS) formation. Electron micrographs indicated increased autophagosome production and endoplasmic reticulum stress, which was supported by positive acridine orange staining and elevated levels of LC3 II, P62 and pIRE1. Screening mTOR signaling revealed a release of inhibition of autophagy. Patient hTEC produced and secreted elevated amounts of the pro-inflammatory cytokine IL8, which was highly correlated with the acridine orange staining. Summarizing, hTEC of MMAuria patients are characterized by disturbed energy metabolism and ROS production that lead to increased autophagy and IL8 secretion. PMID:26420839

  13. Extremely low frequency electromagnetic field sensitizes cisplatin-resistant human ovarian adenocarcinoma cells via P53 activation.

    Science.gov (United States)

    Baharara, Javad; Hosseini, Nasrin; Farzin, Tayebe Ramezani

    2016-08-01

    In the following study, extremely low frequency electromagnetic fields (EL-EMF) radiation was used to restore sensitivity in the cisplatin-resistant A2780 ovarian cancer cells. For this purpose A2780 cells were treated with different doses of cisplatin and EL-EMF (50 Hz, 200 gauss, and 2 h) alone. Cytotoxicity was the measurement using MTT assay. After calculating IC50 for cisplatin (90 µg/ml) a lower concentration from IC50 (30 and 60 µg/ml) was used to be combined with EL-EMF. We compare the effects of each cisplatin, EL-EMF and combination groups using acridine orange-propidium iodide (AO/PI) and DAPI staining, caspase 3/9 activation assay and Annexin/PI assay. We also assessed changes in P53 and Matrix metalloproteinases 2 (MMPs) gene expression with semi-quantitative RT-PCR. Results indicated an EL-EMF-dependent proliferative decrease which was found <10 %, and occurred independently of cisplatin. The decreased proliferation rate for 30 and 60 µg/ml cisplatin was about 20 and 40 %, respectively, while for synergistic groups 30 and 60 µg/ml cisplatin with 2 h EL-EMF exposer, showed 47 and 71 % decrease in viability in rats. DAPI staining indicated that chromatin break down significantly increased in synergistic groups. Acridine orange staining also confirmed MTT assay results. Caspase activity significantly increased in the combined groups. Semi-quantitative RT-PCR showed that in synergistic groups of cisplatin and EL-EMF, expression of P53 was increased but the expression level of MPP-2 gene decreased. Results from this study showed that changes generated by the non-invasive EL-EMF can make resistant cells sensitive to cisplatin. PMID:26370097

  14. Computer-assisted laser scanning and video microscopy for analysis of Cryptosporidium parvum oocysts in soil, sediment, and feces

    Energy Technology Data Exchange (ETDEWEB)

    Anguish, L.J.; Ghiorse, W.C. [Cornell Univ., Ithaca, NY (United States)

    1997-02-01

    A computer-assisted laser scanning microscope equipped for confocal laser scanning and color video microscopy was used to examine Cryptosporidium parvum oocysts in two agricultural soils, a barnyard sediment, and calf fecal samples. An agar smear technique was developed for enumerating oocysts in soil and barnyard sediment samples. Enhanced counting efficiency and sensitivity (detection limit, 5.2 x 10{sup 2} oocysts {center_dot} g [dry weight]{sup -1}) were achieved by using a semiautomatic counting procedure and confocal laser scanning microscopy to enumerate immunostained oocysts and fragments of oocysts in the barnyard sediment. An agarose-acridine orange mounting procedure was developed for high-resolution confocal optical sectioning of oocysts in soil. Stereo images of serial optical section revealed the three-dimensional spatial relationships between immunostained oocysts and the acridine orange-stained soil matrix material. In these hydrated, pyrophosphate-dispersed soil preparations, oocysts were not found to be attached to soil particles. A fluorogenic dye permeability assay for oocyst viability was modified by adding an immunostaining step after application of the fluorogenic dyes propidium iodide and 4{prime},6-diamidino-2-phenylindole. Comparison of conventional color epifluorescence and differential interference contrast images on one video monitor with comparable black-and-white laser-scanned confocal images on a second monitor allowed for efficient location and interpretation of fluorescently stained oocysts in the soil matrix. This multi-imaging procedure facilitated the interpretation of the viability assay results by overcoming the uncertainties caused by matrix interference and background fluorescence. 48 refs., 4 figs., 1 tab.

  15. Interference of PAHs and their N-heterocyclic analogs with signaling of retinoids in vitro.

    Science.gov (United States)

    Benísek, Martin; Bláha, Ludek; Hilscherová, Klára

    2008-12-01

    Retinoids are dietary hormones acting through nuclear receptors for retinoic acid, important especially during embryonic development. This study focuses on the disruption of signaling pathways of retinoids by polycyclic aromatic hydrocarbons (PAHs) and their N-heterocyclic analogs (N-PAHs), important environmental contaminants with numerous biological effects. In vitro test with P19/A15 cell line stably transfected with luciferase reporter gene under control of retinoic acid-responsive elements was used to investigate both direct activation of retinoic acid receptors and modulation of response induced by natural ligand all-trans retinoic acid (ATRA) by 26 PAHs and N-PAHs. While none of individual compounds alone activated retinoic acid receptors, many of them modulated ATRA-mediated activity both after 6 h and 24 h exposure. Majority of compounds active after 6h downregulated ATRA-mediated activity (most effective were two analogs of dibenz[a,h]anthracene with LOECs about 185 nM), while most compounds active after 24h upregulated the effects of ATRA (most effective benz[a]acridine and dibenz[a,i]acridine caused 400% induction of ATRA response). Quantitative structure-activity relationship analysis identified molecular volume and dipole moment as the most important descriptors of inhibitory effects after 6h, while length, total molecular energy, gap-HOMO/LUMO and Van der Waals energy are important descriptors for stimulatory effects of PAHs and N-PAHs. This study demonstrates those abundant pollutants such as PAHs and their analogs interfere in vitro with retinoid signaling, which could play role in some in vivo effects of these organic contaminants such as teratogenicity. PMID:18835432

  16. Development of potent autophagy inhibitors that sensitize oncogenic BRAF V600E mutant melanoma tumor cells to vemurafenib.

    Science.gov (United States)

    Goodall, Megan L; Wang, Tong; Martin, Katie R; Kortus, Matthew G; Kauffman, Audra L; Trent, Jeffrey M; Gately, Stephen; MacKeigan, Jeffrey P

    2014-06-01

    Autophagy is a dynamic cell survival mechanism by which a double-membrane vesicle, or autophagosome, sequesters portions of the cytosol for delivery to the lysosome for recycling. This process can be inhibited using the antimalarial agent chloroquine (CQ), which impairs lysosomal function and prevents autophagosome turnover. Despite its activity, CQ is a relatively inadequate inhibitor that requires high concentrations to disrupt autophagy, highlighting the need for improved small molecules. To address this, we screened a panel of antimalarial agents for autophagy inhibition and chemically synthesized a novel series of acridine and tetrahydroacridine derivatives. Structure-activity relationship studies of the acridine ring led to the discovery of VATG-027 as a potent autophagy inhibitor with a high cytotoxicity profile. In contrast, the tetrahydroacridine VATG-032 showed remarkably little cytotoxicity while still maintaining autophagy inhibition activity, suggesting that both compounds act as autophagy inhibitors with differential effects on cell viability. Further, knockdown of autophagy-related genes showed no effect on cell viability, demonstrating that the ability to inhibit autophagy is separate from the compound cytotoxicity profiles. Next, we determined that both inhibitors function through lysosomal deacidification mechanisms and ultimately disrupt autophagosome turnover. To evaluate the genetic context in which these lysosomotropic inhibitors may be effective, they were tested in patient-derived melanoma cell lines driven by oncogenic BRAF (v-raf murine sarcoma viral oncogene homolog B). We discovered that both inhibitors sensitized melanoma cells to the BRAF V600E inhibitor vemurafenib. Overall, these autophagy inhibitors provide a means to effectively block autophagy and have the potential to sensitize mutant BRAF melanomas to first-line therapies.

  17. Multimodal confocal mosaics enable high sensitivity and specificity in screening of in situ squamous cell carcinoma

    Science.gov (United States)

    Grados Luyando, Maria del Carmen; Bar, Anna; Snavely, Nicholas; Jacques, Steven; Gareau, Daniel S.

    2014-02-01

    Screening cancer in excision margins with confocal microscopy may potentially save time and cost over the gold standard histopathology (H and E). However, diagnostic accuracy requires sufficient contrast and resolution to reveal pathological traits in a growing set of tumor types. Reflectance mode images structural details due to microscopic refractive index variation. Nuclear contrast with acridine orange fluorescence provides enhanced diagnostic value, but fails for in situ squamous cell carcinoma (SCC), where the cytoplasm is important to visualize. Combination of three modes [eosin (Eo) fluorescence, reflectance (R) and acridine orange (AO) fluorescence] enable imaging of cytoplasm, collagen and nuclei respectively. Toward rapid intra-operative pathological margin assessment to guide staged cancer excisions, multimodal confocal mosaics can image wide surgical margins (~1cm) with sub-cellular resolution and mimic the appearance of conventional H and E. Absorption contrast is achieved by alternating the excitation wavelength: 488nm (AO fluorescence) and 532nm (Eo fluorescence). Superposition and false-coloring of these modes mimics H and E, enabling detection of the carcinoma in situ in the epidermal layer The sum mosaic Eo+R is false-colored pink to mimic eosins' appearance in H and E, while the AO mosaic is false-colored purple to mimic hematoxylins' appearance in H and E. In this study, mosaics of 10 Mohs surgical excisions containing SCC in situ and 5 containing only normal tissue were subdivided for digital presentation equivalent to 4X histology. Of the total 16 SCC in situ multimodal mosaics and 16 normal cases presented, two reviewers made 1 and 2 (respectively) type-2 errors (false positives) but otherwise scored perfectly when using the confocal images to screen for the presence of SCC in situ as compared to the gold standard histopathology. Limitations to precisely mimic H and E included occasional elastin staining by AO. These results suggest that

  18. Induction of Apoptosis by Green Synthesized Gold Nanoparticles Through Activation of Caspase-3 and 9 in Human Cervical Cancer Cells

    Science.gov (United States)

    Baharara, Javad; Ramezani, Tayebe; Divsalar, Adeleh; Mousavi, Marzieh; Seyedarabi, Arefeh

    2016-01-01

    Background: Gold Nanoparticles (GNPs) are used in imaging and molecular diagnostic applications. As the development of a novel approach in the green synthesis of metal nanoparticles is of great importance and a necessity, a simple and safe method for the synthesis of GNPs using plant extracts of Zataria multiflora leaves was applied in this study and the results on GNPs’ anticancer activity against HeLa cells were reported. Methods: The GNPs were characterized by UV-visible spectroscopy, FTIR, TEM, DLS and Zeta-potential measurements. In addition, the cellular up-take of nanoparticles was investigated using Dark Field Microscopy (DFM). Induction of apoptosis by high dose of GNPs in HeLa cells was assessed by MTT assay, Acridin orange, DAPI staining, Annexin V/PI double-labeling flow cytometry and caspase activity assay. Results: UV-visible spectroscopy results showed a surface plasmon resonance band for GNPs at 530 nm. FTIR results demonstrated an interaction between plant extract and nanoparticles. TEM images revealed different shapes for GNPs and DLS results indicated that the GNPs range in size from 10 to 42 nm. The Zeta potential values of the synthesized GNPs were between 30 to 50 Mev, indicating the formation of stable particles. As evidenced by MTT assay, GNPs inhibit proliferation of HeLa cells in dose-dependent GNPs and cytotoxicity of GNPs in Bone Marrow Mesenchymal Stem Cell (BMSCs) was lower than cancerous cells. At nontoxic concentrations, the cellular up-take of the nanoparticles took place. Acridin orange and DAPI staining showed morphological changes in the cell’s nucleus due to apoptosis. Finally, caspase activity assay demonstrated HeLa cell’s apoptosis through caspase activation. Conclusion: The results showed that GNPs have the ability to induce apoptosis in HeLa cells. PMID:27141266

  19. β-Hydroxybutyrate supports synaptic vesicle cycling but reduces endocytosis and exocytosis in rat brain synaptosomes.

    Science.gov (United States)

    Hrynevich, Sviatlana V; Waseem, Tatyana V; Hébert, Audrey; Pellerin, Luc; Fedorovich, Sergei V

    2016-02-01

    The ketogenic diet is used as a prophylactic treatment for different types of brain diseases, such as epilepsy or Alzheimer's disease. In such a diet, carbohydrates are replaced by fats in everyday food, resulting in an elevation of blood-borne ketone bodies levels. Despite clinical applications of this treatment, the molecular mechanisms by which the ketogenic diet exerts its beneficial effects are still uncertain. In this study, we investigated the effect of replacing glucose by the ketone body β-hydroxybutyrate as the main energy substrate on synaptic vesicle recycling in rat brain synaptosomes. First, we observed that exposing presynaptic terminals to nonglycolytic energy substrates instead of glucose did not alter the plasma membrane potential. Next, we found that synaptosomes were able to maintain the synaptic vesicle cycle monitored with the fluorescent dye acridine orange when glucose was replaced by β-hydroxybutyrate. However, in presence of β-hydroxybutyrate, synaptic vesicle recycling was modified with reduced endocytosis. Replacing glucose by pyruvate also led to a reduced endocytosis. Addition of β-hydroxybutyrate to glucose-containing incubation medium was without effect. Reduced endocytosis in presence of β-hydroxybutyrate as sole energy substrate was confirmed using the fluorescent dye FM2-10. Also we found that replacement of glucose by ketone bodies leads to inhibition of exocytosis, monitored by FM2-10. However this reduction was smaller than the effect on endocytosis under the same conditions. Using both acridine orange in synaptosomes and the genetically encoded sensor synaptopHluorin in cortical neurons, we observed that replacing glucose by β-hydroxybutyrate did not modify the pH gradient of synaptic vesicles. In conclusion, the nonglycolytic energy substrates β-hydroxybutyrate and pyruvate are able to support synaptic vesicle recycling. However, they both reduce endocytosis. Reduction of both endocytosis and exocytosis together with

  20. Spectrophotometric Determination of Trace Amount of Iodide with Chromium(Ⅵ )- Bromide- Basic Xanthene Dye System%痕量碘离子的铬 (Ⅵ)-碱性吨染料体系分光光度法测定

    Institute of Scientific and Technical Information of China (English)

    吕明玉; 刘绍璞

    2001-01-01

    在过量溴化物存在下的稀磷酸介质中, I-被 Cr(Ⅵ)氧化成 I2后与 Br-结合形成 [I2Br]-配阴离子,该配阴离子能进一步与罗丹明 6G、罗丹明 B、吖啶红等碱性吨染料阳离子形成离子缔合配合物。在聚乙烯醇存在下,缔合物体系稳定且溶液颜色有明显的变化,可用于 I-离子的光度测定。方法具有高灵敏度,不同体系的摩尔吸光系数在 4. 96× 104~ 1. 1× 105 L· mol- 1· cm- 1之间,以罗丹明 6G和罗丹明 B体系灵敏度较高。碘离子质量浓度分别在 0~ 0.8 mg/L(罗丹明 B和罗丹明 6G体系 )、 0~ 1.0 mg/L(吖啶红体系)之间遵守比尔定律。方法具有良好的选择性,用于海带、黄豆和含碘药片的测定结果令人满意。%In dilute phosphoric acid medium, chromium(Ⅵ ) oxidizes I-to I2, which then binds Br-to form [I2Br]-anion in the presence of excess bromide, and [I2Br]-anion further reacts with basic xanthene dyes such as rhodamine 6G, rhodamine B and acridine red to form ion -association complexes. In the presence of polyvinyl alcohol(PVA) the solution is stable and a distinct color change occurred. The method can be applied to the spectrophotometric determination of iodide. The method have a high sensitivity and selectivity. Its molar absorptivity is 1.1× 105 L· mol-1· cm-1 for rhodamine 6G and rhodamine B systems, and 4.96× 104 L· mol-1· cm-1 for acridine red system. The Beer’ s law is obeyed in the range over 0~ 0.8 mg/L and 0~ 1.0 mg/L of I-for rhodamine 6G, B systems, and acridine red system, respectively. It was used to determine I-in kelp, soybean and cydiodine tablets with satisfactory results.

  1. Detection of Extermination Effect of Ultraviolet Irradiation on Gypsy Moth Eggs by Fluorescence Probe Technique%紫外线照射灭卵效果的荧光探针技术检测

    Institute of Scientific and Technical Information of China (English)

    李景奎; 戚大伟

    2011-01-01

    以林木害虫舞毒蛾(Lymantria dispar L.)虫卵为试验材料,利用吖啶橙、罗丹明123、Ho33342和碘化丙啶4种荧光探针追踪标记虫卵细胞内部的不同细胞器,研究了紫外线照射对舞毒蛾虫卵的微现影响.研究表明:经过紫外线照射后,吖啶橙标记的虫卵细胞产生红色荧光;罗丹明123标记虫卵细胞荧光强度有所减弱;Ho33342和碘化丙啶标记的虫卵细胞产生大量粉红色荧光,并且,细胞形态发生了改变.紫外线照射导致虫卵细胞大量DNA链断裂,线粒体膜电位差下降,坏死细胞逐渐增多.荧光探针技术体现了物理灭虫的有效性.%An experiment was conducted to study the effect of ultraviolet irradiation on the microstructure of gypsy moth (Lymantria dispar L. ) eggs using fluorescence probes such as acridine orange, rhodanmine123, Ho33342 and Propidium iodide (PI) to trace and mark different organelles in egg cells. Results indicated that, after ultraviolet in'adiation, the egg cells marked with acridine orange gave off red fluorescence; the fluorescence intensity of egg cells marked with rhodamine123 decreased; egg cells marked with Ho33342 and PI gave off a lot of pink fluorescence, and the shape of cells changed. Because of the ultraviolet irradiation,much DNA of egg cells strands broke; electric potential of mitochondria membrane decreased; necrotic cells increased by degrecs. Fluorescence probe technique shows the validity of physical extermination of insects.

  2. In vitro effects of polyphenols on colorectal cancer cells

    Institute of Scientific and Technical Information of China (English)

    Barbara; Pampaloni; Gaia; Palmini; Carmelo; Mavilia; Roberto; Zonefrati; Annalisa; Tanini; Maria; Luisa; Brand

    2014-01-01

    AIM:To investigate the effects of quercetin and genistein on colon cancer cell proliferation and their estrogen receptorβ(ERβ)expression.METHODS:Colon cancer cells were stably transfected with a mammalian expression vector to overexpress ERβ(HCT8-β8-expressing cells)or a control vector(HCT8-pSV2neo-expressing cells).The proliferation of these cells was examined after treatment with quercetin or genistein(5-100μmol/L),or 10 nmol/L17β-estradiol(17β-E2).Cell viability was examined by acridine orange staining following treatments for 48 or144 h.Effects of quercetin and genistein on ERβtranscriptional transactivation were examined by luciferase activity in HCT8-β8-expressing cells transiently transfected with a pEREtkLUC reporter vector.In addition,the regulation of ERβtranscription by phytoestrogens and 17β-E2 was examined by quantitative polymerase chain reaction.RESULTS:Proliferation of HCT8-β8-expressing cells was not reduced low doses(5μmol/L)of quercetin and genistein,while it was reduced at 25-50μmol/L with an effect similar to 10 nmol/L 17β-E2.Treatment with doses of phytoestrogens≥75μmol/L completely blocked cell growth and reduced overall cell counts,however no effects at any dose were observed in HCT8-pSV2neoexpressing cells.These results were supported by viability staining that revealed acridine orange-stained lysosomes with high doses or extended treatment periods.Genistein and quercetin(50μmol/L)significantly increased ER-responsive luciferase activity similar to 10nmol/L 17β-E2(P<0.05).Furthermore,genistein and quercetin(50μmol/L),as well as 10 nmol/L 17β-E2significantly increased ERβmRNA levels in HCT8-β8-expressing cells(P<0.05).In addition,treatment of HCT8-pSV2neo-expressing cells with 50μmol/L quercetin or 10 nmol/L 17β-E2 significantly increased ERβmRNA levels compared to untreated controls(P<0.05),though the absolute levels were much lower than in HCT8-β8-expressing cells.CONCLUSION:The antitumorigenic effects of the

  3. The Sperm Chromatin Structure Assay (SCSA(®)) and other sperm DNA fragmentation tests for evaluation of sperm nuclear DNA integrity as related to fertility.

    Science.gov (United States)

    Evenson, Donald P

    2016-06-01

    Thirty-five years ago the pioneering paper in Science (240:1131) on the relationship between sperm DNA integrity and pregnancy outcome was featured as the cover issue showing a fluorescence photomicrograph of red and green stained sperm. The flow cytometry data showed a very significant difference in sperm DNA integrity between fertile and subfertile bulls and men. This study utilized heat (100°C, 5min) to denature DNA at sites of DNA strand breaks followed by staining with acridine orange (AO) and measurements of 5000 individual sperm of green double strand (ds) DNA and red single strand (ss) DNA fluorescence. Later, the heat protocol was changed to a low pH protocol to denature the DNA at sites of strand breaks; the heat and acid procedures produced the same results. SCSA data are very advantageously dual parameter with 1024 channels (degrees) of both red and green fluorescence. Hundreds of publications on the use of the SCSA test in animals and humans have validated the SCSA as a highly useful test for determining male breeding soundness. The SCSA test is a rapid, non-biased flow cytometer machine measurement providing robust statistical data with exceptional precision and repeatability. Many genotoxic experiments showed excellent dose response data with very low coefficient of variation that further validated the SCSA as being a highly powerful assay for sperm DNA integrity. Twelve years following the introduction of the SCSA test, the terminal deoxynucleotidyl transferase-mediated fluorescein-dUTP nick end labelling (TUNEL) test (1993) for sperm was introduced as the only other flow cytometric assay for sperm DNA fragmentation. However, the TUNEL test can also be done by light microscopy with much less statistical robustness. The COMET (1998) and Sperm Chromatin Dispersion (SCD; HALO) (2003) tests were introduced as light microscope tests that don't require a flow cytometer. Since these tests measure only 50-200 sperm per sample, they suffer from the lack of

  4. Synthesis, structural characterization, and anticancer activity of a monobenzyltin compound against MCF-7 breast cancer cells

    Directory of Open Access Journals (Sweden)

    Fani S

    2015-11-01

    Full Text Available Somayeh Fani,1 Behnam Kamalidehghan,1 Kong Mun Lo,2 Najihah Mohd Hashim,1 Kit May Chow,2 Fatemeh Ahmadipour1 1Department of Pharmacy, Faculty of Medicine, 2Department of Chemistry, Faculty of Science, University of Malaya, Kuala Lumpur, Malaysia Abstract: A new monoorganotin Schiff base compound, [N-(3,5-dichloro-2-oxidobenzylidene-4-chlorobenzyhydrazidato](o-methylbenzylaquatin(IV chloride, (compound C1, was synthesized, and its structural features were investigated by spectroscopic techniques and single-crystal X-ray diffractometry. Compound C1 was exposed to several human cancer cell lines, including breast adenocarcinoma cell lines MCF-7 and MDA-MB-231, ovarian adenocarcinoma cell lines Skov3 and Caov3, and prostate cancer cell line PC3, in order to examine its cytotoxic effect for different forms of cancer. Human hepatic cell line WRL-68 was used as a normal cell line. We concentrated on the MCF-7 cell line to detect possible underlying mechanism involvement of compound C1. 3-(4,5-dimethylthiazol-2-yl-2,5-diphenyltetrazolium bromide (MTT assay revealed the strongest cytotoxicity of compound C1 against MCF-7 cells, with a half maximal inhibitory concentration (IC50 value of 2.5±0.50 µg/mL after 48 hours treatment. The IC50 value was >30 µg/mL in WRL-68 cells. Induced antiproliferative activity of compound C1 for MCF-7 cells was further confirmed by lactate dehydrogenase, reactive oxygen species, acridine orange/propidium iodide staining, and DNA fragmentation assays. A significant increase of lactate dehydrogenase release in treated cells was observed via fluorescence analysis. Luminescent analysis showed significant growth in intracellular reactive oxygen species production after treatment. Morphological changes of necrosis and early and late apoptosis stages were observed in treated cells after staining with acridine orange/propidium iodide. DNA fragmentation was observed as a characteristic of apoptosis in treated cells. Results of the

  5. Using Different Staining Agents to Identify the Living Status of Circulating Hemocytes in the Silkworm, Bombyx mori%利用不同染料复合染色判定家蚕血细胞的存活状况

    Institute of Scientific and Technical Information of China (English)

    李旭全; 杨兵; 路岸瑞; 刘朝良; 凌尔军

    2011-01-01

    Bombyx mori has 5 types of circulating hemocytes that can be clearly identified after being stained by acridine orange (AO) and propidium iodide (PI). 3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyl-2H-tetrazolium bromide (MTT) is another type of staining agent that has been extensively used to study cell proliferation and to identify living cells. Bombyx mori hemocytes undergone multiple staining, I. E. Staining with MTT first and then with AO and PI, showed two patterns. One was that a majority of granulocytes and plasmatocytes and all prohemocytes could not be stained by MTT but were shown to be living cells from AO staining. The other was that the MTT stained hemocytes had a lot of purple fiber-like structure extending from cell surface. All MTT-stained hemocytes but some granulocytes with pseudopods could also be stained by PI according to their nuclei with red fluorescence. Therefore, careful analysis and discriminative treatment should be conducted when multiple staining with these three staining agents are used to tell whether a cell is living or dead.%家蚕有5种血细胞,可以通过吖啶橙(acridine orange,AO)和碘化丙啶(propidium iodide,PI)染色来区分.溴化噻唑蓝四氮唑[3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyl-2 H-tetrazolium bromide,MTT]是一种广泛应用于细胞增殖和活细胞鉴定的染色剂.通过对家蚕血细胞进行复染,即先用MTT染色,再用AO和PI染色,发现以下2种情况:很多颗粒细胞、浆细胞和所有的原血球细胞不能被MTT染色,但AO染色显示为活的细胞;被MTT染色的细胞表面有紫色纤维状物伸出,除了一部分具有伪足的颗粒细胞外,所有被MTT染色的血细胞也被PI染色,细胞核是红色.因此,利用这3种染料对家蚕血细胞复合染色来判断细胞的存活与死亡,需要仔细分析并分别对待.

  6. Preparation of arsenic trioxide-loaded albuminutes immuno-nanospheres and its specific killing effect on bladder cancer cell in vitro

    Institute of Scientific and Technical Information of China (English)

    ZHOU Jie; ZENG Fu-qing; LI Chong; TONG Qiang-song; GAO Xiang; XIE Shu-sheng; YU Li-zhang

    2005-01-01

    Background Recently, arsenic trioxide (As2O3) was considered as a novel anti-tumor agent. However, it showed severe toxicity effect on normal tissue at the same time. To improve its therapeutic efficacy and decrease its toxicity,we prepared arsenic trioxide-loaded albuminutes immuno-nanospheres [As2O3-(HAS-NS)-BDI-1] targeted with nonoclonal antibody (McAb) BDI-1 and tested its specific killing effect against bladder cancer cell. Methods As2O3-HAS-NS was prepared by chemical cross-linking method. Monoclonal antibody BDI-1 was purified with ammonium sulphate saltingout and chromatography. Albuminutes microspheres were conjugated with McAb by SPDP cross-linking method.Concentration of As in As2O3- (HAS-NS)-BDI-1 and As2O3-HAS-NS was measured by atomic fluometry method. As2O3- (HAS-NS)-BDI-1 and its activity were detected by SDS-PAGE reduction electrophoresis, indirect immunofluorescence test, light microscope and scanning electron microscope observation. Acridine orange staining and tritiated thymidine (3H-TdR) incorporation tests were used to indicate specific killing activity of As2O3-(HAS-NS)-BDI-1 in vitro. Results In As2O3- (HAS-NS)-BDI-1 groups, we saw two protein bands in SDS-PAGE reduction electrophoresis.Albuminutes immuno-nanospheres were rounded with clear green fluorescence by immunofluorescence test. Under microscope, we observed that BIU-87 cells were covered with the As2O3- (HAS-NS)-BDI-1 and that As2O3- (HAS-NS)-BDI-1 moved with the BIU-87 cells. The albuminutes immuno-nanospheres were tightly junctioned with the BIU-87 cells.Specific killing activity of As2O3-(HAS-NS)-BDI-1 on bladder tumor cells was observed by acridine orange staining and 3H-TdR incorporation assays. Conclusions As2O3- (HAS-NS)-BDI-1 might bind specifically against BIU-87 cells, thus leading to high activity of killing bladder tumor cells.

  7. Antineoplastic effects of deoxyelephantopin,a sesquiterpene lactone from Elephantopus scaber, on lung adenocarcinoma (A549) cells

    Institute of Scientific and Technical Information of China (English)

    Farha A.Kabeer; Geetha B.Sreedevi; Mangalam S.Nair; Dhanya S.Rajalekshmi; Latha P.Gopalakrishnan; Sujathan Kunjuraman; Remani Prathapan

    2013-01-01

    OBJECTIVE:Deoxyelephantopin,a sesquiterpene lactone from Elephantopus scaber,showed inhibition of the growth of various tumor cells in vitro.In the present study,we investigated the cytotoxicity and apoptosis-inducing capacity of deoxyelephantopin on lung adenocarcinoma (A549) cells.METHODS:The cytotoxic effect of deoxyelephantopin on A549 cells and normal lymphocytes was evaluated using 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay and 50% inhibitory concentration (IC50) value was determined.The self-renewal and proliferating potential of A549 cells after treatment with deoxyelephantopin were examined by colony formation assay.Cellular morphology of deoxyelephantopin-treated cells was observed using phasecontrast microscopy.The induction of apoptosis was evaluated using acridine orange and ethidium bromide staining,Hoechst 33342 staining,terminal deoxynucleotidyl transferase-mediated dUTP biotin nick end-labeling (TUNEL) assay,DNA fragmentation analysis and Annexin V-fluorescein isothiocyanate staining by flow cytometry.Activation of caspases was detected using fluorogenic substrate specific to caspases 2,3,8 and 9 and flow cytometric analysis.The total cellular DNA content and expression of cleaved poly (ADP-ribose) polymerase was also analyzed.RESULTS:Deoxyelephantopin exhibited cytotoxicity to A549 cells (IC5o =12.287 μg/mL),however,there was no toxicity towards normal human lymphocytes.Deoxyelephantopin suppressed the colony-forming ability of A549 cells in a dose-dependent manner.Acridine orange,ethidium bromide and Hoechst 33342 staining showed cell shrinkage,chromosomal condensation and nuclear fragmentation,indicating induction of apoptosis.Deoxyelephantopin increased apoptosis of A549 cells,as evidenced by more TUNEL-positive cells.DNA fragmentation and Annexin V staining revealed late-stage apoptotic cell population.Deoxyelephantopin inhibited A549 cell growth by cell cycle arrest at G2/M phase and induced apoptosis through

  8. Polimixina B: efeito dose e tempo dependente na nefrotoxicidade in vitro Polymyxin B: dose and time dependent nephrotoxicity effect in vitro

    Directory of Open Access Journals (Sweden)

    Luciana Barros de Moura Neiva

    2013-01-01

    Full Text Available OBJETIVO: Caracterizar a toxicidade da polimixina B (PmxB em células renais em dosagem e tempos diferentes. MÉTODOS: Células LLC-PK1, cultivadas em placas multiwell de 12 poços, foram divididas nos seguintes grupos: Controle (CTL - células mantidas em meio DMEM suplementado a 5%; G1 - células expostas à concentração de 75mM de PmxB; G2 - células expostas à concentração de 375mM de PmxB. Cada grupo foi avaliado nos tempos de 24, 48 e 72 horas quanto à viabilidade celular (Acridine Orange/Brometo de Etídio e apoptose (Hoechst 33342. RESULTADOS: Os dados demonstraram a viabilidade celular e a apoptose à exposição de três doses de PmxB em três intervalos de tempo, com um aumento significativo da toxicidade à elevação das doses e ao maior tempo de permanência no antibiótico para apoptose. CONCLUSÃO: A citotoxicidade pela PmxB, no modelo de cultivo celular, se mostrou tempo e dose dependente, aumentando com a maior exposição e maior dose de antibiótico.OBJECTIVE: To characterize the toxicity of polymyxin B (PmxB in renal cell in different dosage and times. METHODS: LLC-PK1 cells grown in 12 well multiwell plates were divided into the following groups: Control (CTL - cells maintained in DMEM supplemented with 5%; G1 - cells exposed to concentration of 75µM PmxB G2 - cells exposed to concentration of 375µM PmxB. Each group was assessed at 24,48 and 72 hours as for cell viability (Acridine orange/ethidium bromide and apoptosis (Hoechst 33342. RESULTS: The data demonstrate the cell viability and apoptosis exposure of three doses of PmxB in three time intervals, with a significant increase in toxicity to high doses and longer duration of stay in the antibiotic to apoptosis. CONCLUSION: Cytotoxicity by PmxB in cell culture model, showed to be time and dose dependent, increasing with increased exposure and higher dose of antibiotic.

  9. Clinical applications of in vivo fluorescence confocal laser scanning microscopy

    Science.gov (United States)

    Oh, Chilhwan; Park, Sangyong; Kim, Junhyung; Ha, Seunghan; Park, Gyuman; Lee, Gunwoo; Lee, Onseok; Chun, Byungseon; Gweon, Daegab

    2008-02-01

    Living skin for basic and clinical research can be evaluated by Confocal Laser Scanning Microscope (CLSM) non-invasively. CLSM imaging system can achieve skin image its native state either "in vivo" or "fresh biopsy (ex vivo)" without fixation, sectioning and staining that is necessary for routine histology. This study examines the potential fluorescent CLSM with a various exogenous fluorescent contrast agent, to provide with more resolution images in skin. In addition, in vivo fluorescent CLSM researchers will be extended a range of potential clinical application. The prototype of our CLSM system has been developed by Prof. Gweon's group. The operating parameters are composed of some units, such as illuminated wavelength 488 nm, argon illumination power up to 20mW on the skin, objective lens, 0.9NA oil immersion, axial resolution 1.0μm, field of view 200μm x 100μm (lateral resolution , 0.3μm). In human volunteer, fluorescein sodium was administrated topically and intradermally. Animal studies were done in GFP transgenic mouse, IRC mouse and pig skin. For imaging of animal skin, fluorescein sodium, acridine orange, and curcumine were used for fluorescein contrast agent. We also used the GFP transgenic mouse for fluorescein CLSM imaging. In intact skin, absorption of fluorescein sodium by individual corneocyte and hair. Intradermal administrated the fluorescein sodium, distinct outline of keratinocyte cell border could be seen. Curcumin is a yellow food dye that has similar fluorescent properties to fluorescein sodium. Acridin Orange can be highlight nuclei in viable keratinocyte. In vivo CLSM of transgenic GFP mouse enable on in vivo, high resolution view of GFP expressing skin tissue. GFP signals are brightest in corneocyte, kertinocyte, hair and eccrine gland. In intact skin, absorption of fluorescein sodium by individual corneocyte and hair. Intradermal administrated the fluorescein sodium, distinct outline of keratinocyte cell border could be seen. In

  10. Design and choice of TFO binding and cleaving HBV core promoter

    Institute of Scientific and Technical Information of China (English)

    光丽霞; 袁发焕; 任平; 奚敏; 艾友平

    2003-01-01

    Objective: To screen a triple helix-forming oligodeoxyribonucleotide (TFO) that can bind HBV core promoter at target site with high affinity and specificity, and to observe the ability of manganese porphyrin modified TFO to combine and cleave HBV DNA.Methods: Similar homopurine domain (1 734-1 754) in HBV core promoter was selected as target sequence.Several corresponding TFOs were synthesized.The affinities and specificities of TFOs binding target sequence were tested with electrophoretic mobility shift and DNase Ⅰ footprinting assays.The selected best TFO was modified with manganese porphyrin and acridine.The ability of the TFO derivative to cleave HBV DNA was observed with cleavage experiment.Results: Under the condition of 37℃ and pH 7.4, the TFO consisting of cytidylate and thymidylate (CT-TFO) and the parallel TFO consisting of guanylate and thymidylate (GT-TFOp) bound the target sequence weakly with Kd values much more than 10-6 mol/L.The affinities of anti-parallel GT-TFO (GT-TFOap) and short TFO consisting of adenine nucleotide and guanylate (AG-TFOsh) binding the target sequence were higher than those of the formers, with Kd values of 5×10-7 mol/L and 2.5×10-8 mol/L respectively.Long AG-TFO (AG-TFOl) had the highest binding affinity with a Kd value of 3×10-9 mol/L among all the TFOs studied for sequence specificity.In the presence of potassium monopersulfate, KHSO5, TFO modified with manganese porphyrin and acridine cleaved the target sequence where the triplex DNA formed.Conclusion: TFO containing AG or GT binds homopurine in HBV core promoter in adverse parallel direction to form triple helix.AG-TFOl has the highest binding affinity among all the TFOs studied.After modified with manganese porphyrin, AG-TFOl completely binds and cleaves the target HBV DNA sequence where triplex DNA is formed.

  11. Novel microfilaricidal activity of nanosilver

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    Singh SK

    2012-02-01

    Full Text Available Sunil K Singh1, Kalyan Goswami2, Richa D Sharma2, Maryada VR Reddy2, Debabrata Dash11Department of Biochemistry, Institute of Medical Sciences, Banaras Hindu University, Varanasi, 2Department of Biochemistry, Mahatma Gandhi Institute of Medical Sciences, Sevagram, IndiaPurpose: The currently available drug repertoire against lymphatic filariasis, a major health hazard in the developing world, is inadequate and is fraught with serious limitations. Thus, the development of an effective antifilarial strategy has become a global research thrust mandated by the World Health Organization. Nanoparticles of silver endowed with antibacterial potency are known to induce apoptosis in eukaryotic cells. The present study was designed to investigate the possible microfilaricidal efficacy of silver nanoparticles and to establish the validity of apoptotic rationale in antifilarial drug designing.Methods: This report analyzed the effect of nanoparticles of silver as well as gold (size range: 10–15 nm on the microfilariae of Brugia malayi obtained from the lavage of peritoneal cavities of infected jirds (Meriones unguiculatus. The study included a microfilarial motility assay, a trypan blue exclusion test, a poly(adenosine diphosphate-ribose polymerase activity study, ethidium bromide/acridine orange differential staining, and transmission, as well as scanning electron microscopic evaluation of ultrastructural changes in microfilariae.Results: The study demonstrates that nanoparticles of silver, but not of gold, elicited significant loss in microfilarial motility. Differential staining of parasites with ethidium bromide and acridine orange, poly(adenosine diphosphate-ribose polymerase activity in microfilarial lysate, and electron microscopic findings underscored apoptotic death of parasites attributable to nanosilver. In a trypan blue exclusion test, the 50% lethal dose of nanosilver was measured to be 101.2 µM, which was higher than the recorded complete

  12. Modelling toxicity induced Neurological disorders in Zebrafish

    Directory of Open Access Journals (Sweden)

    Benin Joseph

    2012-03-01

    Full Text Available Neurological disorders have become more common and prevalent. Cellular pathology and behavioural symptoms in neurodegenerative diseases although connected are still a mystery to solve with no complete cure available yet. Central pathways in neurodegeneration involves impaired ubiquitin-proteasome machinery, autophagy and mitochondrial oxidative stress. In the case of neurodevlopmental disorders, environmental toxins and genetic factors are main causative agents. We aim to create a toxicity induced zebrafish model of neurological disease focussing on cognition, movement and hyperactivity disorders. Zebra fish embryos at 48 hr post fertilization were treated with different doses of lead, cholesterol and acetyl choline and by 7 days post fertilization pectoral fin movement, swimming behaviour and touch response were compromised in parallel with apoptosis identified in the brain by acridine orange fluorescent staining. A marked window is observed, therefore promising for a drug screening platform. Further characterization of pathology associated protein expression and specific behavioural studies could render this as a simple promising toxic model for preclinical drug screening.

  13. Transformation of the antiepileptic drug oxcarbazepine upon different water disinfection processes.

    Science.gov (United States)

    Li, Zhi; Fenet, Hélène; Gomez, Elena; Chiron, Serge

    2011-02-01

    Transformation of the pharmaceutical oxcarbazepine (OXC), a keto analogue of carbamazepine (CBZ) was investigated under different water disinfection processes (ozonation, chlorination and UV irradiation) to compare its persistence, toxicity and degradation pathways with those of CBZ. Analysis by LC-ion trap-MS(n) allowed for the identification of up to thirteen transformation products (TPs). The major abundant and persistent TPs (10,11-dihydro-10,11-trans-dihydroxy-carbamazepine (DiOH-CBZ), acridine (ACIN) and 1-(2-benzaldehyde)-(1H, 3H)-quinazoline-2,4-dione (BQD)) were identical to those previously reported during water treatment of CBZ. Only one new compound arising from an intramolecular cyclisation reaction was identified during UV irradiation. OXC reacted quickly with hydroxyl radical and relatively rapidly with free chlorine while slow reaction rates were recorded in presence of ozone and upon UV irradiation. An increase of the acute toxicity of UV irradiated solutions, monitored by a Daphnia magna bioassay, was recorded, probably due to the accumulation of ACIN. The formation of ACIN is of concern due to the carcinogenic properties of this chemical. ACIN was also generated during the direct UV photo transformation of DiOH-CBZ and 10-hydroxy-10,11-dihydro-carbamazepine (OH-CBZ), two metabolites of OXC and CBZ widely detected in water resources. Analysis of tap water samples revealed the occurrence at ng/L levels of the major TPs detected under laboratory scale experiments, except ACIN.

  14. A Versatile Cell Death Screening Assay Using Dye-Stained Cells and Multivariate Image Analysis.

    Science.gov (United States)

    Collins, Tony J; Ylanko, Jarkko; Geng, Fei; Andrews, David W

    2015-11-01

    A novel dye-based method for measuring cell death in image-based screens is presented. Unlike conventional high- and medium-throughput cell death assays that measure only one form of cell death accurately, using multivariate analysis of micrographs of cells stained with the inexpensive mix, red dye nonyl acridine orange, and a nuclear stain, it was possible to quantify cell death induced by a variety of different agonists even without a positive control. Surprisingly, using a single known cytotoxic agent as a positive control for training a multivariate classifier allowed accurate quantification of cytotoxicity for mechanistically unrelated compounds enabling generation of dose-response curves. Comparison with low throughput biochemical methods suggested that cell death was accurately distinguished from cell stress induced by low concentrations of the bioactive compounds Tunicamycin and Brefeldin A. High-throughput image-based format analyses of more than 300 kinase inhibitors correctly identified 11 as cytotoxic with only 1 false positive. The simplicity and robustness of this dye-based assay makes it particularly suited to live cell screening for toxic compounds.

  15. VITALITY AND MORPHOLOGY OF TUMOR CELLS TREATED WITH 4-TIAZOLIDINONE DERIVATIVES IMMOBILIZED ON NANOSCALE POLYMER CARRIER

    Directory of Open Access Journals (Sweden)

    N. M. Boiko

    2015-02-01

    Full Text Available A nanoscale polymeric carrier was used for delivery of novel anticancer compounds – 4-tiazolidinone derivatives – to tumor cells of different lines. It was found that such way of delivery of the above mentioned compounds to target cells significantly (approximately 10 times decreased acting cytotoxic dose of some of these compounds with preservation of similar level of their antineoplastic effect in vitro towards various mammalian tumor cells. The microscopic investigation of these cells demonstrated that under the action of some immobilized 4-tiazolidonone derivatives, there was an increase (up to 40% of the part of apoptotic cells, as well as an appearance of 10% of cells with morphologically changed nucleus, and up to 35% of cells with an increased intensity of red fluorescence of acridine orange in the lysosomes, compared with such indicators observed under the action of free form of those compounds. Thus, the applied nanoscale carrier is a perspective polymer system for delivery of anticancer drugs to target cells.

  16. Hydrogenation of CO2 to Formic Acid with a Highly Active Ruthenium Acriphos Complex in DMSO and DMSO/Water.

    Science.gov (United States)

    Rohmann, Kai; Kothe, Jens; Haenel, Matthias W; Englert, Ulli; Hölscher, Markus; Leitner, Walter

    2016-07-25

    The novel [Ru(Acriphos)(PPh3 )(Cl)(PhCO2 )] [1; Acriphos=4,5-bis(diphenylphosphino)acridine] is an excellent precatalyst for the hydrogenation of CO2 to give formic acid in dimethyl sulfoxide (DMSO) and DMSO/H2 O without the need for amine bases as co-reagents. Turnover numbers (TONs) of up to 4200 and turnover frequencies (TOFs) of up to 260 h(-1) were achieved, thus rendering 1 one of the most active catalysts for CO2 hydrogenations under additive-free conditions reported to date. The thermodynamic stabilization of the reaction product by the reaction medium, through hydrogen bonds between formic acid and clusters of solvent or water, were rationalized by DFT calculations. The relatively low final concentration of formic acid obtained experimentally under catalytic conditions (0.33 mol L(-1) ) was shown to be limited by product-dependent catalyst inhibition rather than thermodynamic limits, and could be overcome by addition of small amounts of acetate buffer, thus leading to a maximum concentration of free formic acid of 1.27 mol L(-1) , which corresponds to optimized values of TON=16×10(3) and TOFavg ≈10(3)  h(-1) . PMID:27356513

  17. High-Efficiency Selective Electron Tunnelling in a Heterostructure Photovoltaic Diode.

    Science.gov (United States)

    Jia, Chuancheng; Ma, Wei; Gu, Chunhui; Chen, Hongliang; Yu, Haomiao; Li, Xinxi; Zhang, Fan; Gu, Lin; Xia, Andong; Hou, Xiaoyuan; Meng, Sheng; Guo, Xuefeng

    2016-06-01

    A heterostructure photovoltaic diode featuring an all-solid-state TiO2/graphene/dye ternary interface with high-efficiency photogenerated charge separation/transport is described here. Light absorption is accomplished by dye molecules deposited on the outside surface of graphene as photoreceptors to produce photoexcited electron-hole pairs. Unlike conventional photovoltaic conversion, in this heterostructure both photoexcited electrons and holes tunnel along the same direction into graphene, but only electrons display efficient ballistic transport toward the TiO2 transport layer, thus leading to effective photon-to-electricity conversion. On the basis of this ipsilateral selective electron tunnelling (ISET) mechanism, a model monolayer photovoltaic device (PVD) possessing a TiO2/graphene/acridine orange ternary interface showed ∼86.8% interfacial separation/collection efficiency, which guaranteed an ultrahigh absorbed photon-to-current efficiency (APCE, ∼80%). Such an ISET-based PVD may become a fundamental device architecture for photovoltaic solar cells, photoelectric detectors, and other novel optoelectronic applications with obvious advantages, such as high efficiency, easy fabrication, scalability, and universal availability of cost-effective materials.

  18. Calcium uptake and proton transport by acidocalcisomes of Toxoplasma gondii.

    Directory of Open Access Journals (Sweden)

    Peter Rohloff

    Full Text Available Acidocalcisomes are acidic calcium stores found in diverse organisms, being conserved from bacteria to humans. They possess an acidic matrix that contains several cations bound to phosphates, which are mainly present in the form of short and long polyphosphate chains. Their matrix is acidified through the action of proton pumps such as a vacuolar proton ATPase and a vacuolar proton pyrophosphatase. Calcium uptake occurs through a Ca(2+/H(+ countertransporting ATPase located in the membrane of the organelle. Acidocalcisomes have been identified in a variety of microorganisms, including Apicomplexan parasites such as Plasmodium and Eimeria species, and in Toxoplasma gondii. We report the purification and characterization of an acidocalcisome fraction from T. gondii tachyzoites after subcellular fractionation and further discontinuous iodixanol gradient purification. Proton and calcium transport activities in the fraction were characterized by fluorescence microscopy and spectrophotometric methods using acridine orange and arsenazo III, respectively. This work will facilitate the understanding of the function of acidocalcisomes in Apicomplexan parasites, as we can now isolate highly purified fractions that could be used for proteomic analysis to find proteins that may clarify the biogenesis of these organelles.

  19. Comparison of different diagnostic techniques in Plasmodium falciparum cerebral malaria

    Directory of Open Access Journals (Sweden)

    Fatima Shujatullah, Abida Malik, Haris M. Khan & Ashraf Malik

    2006-12-01

    Full Text Available Background & objectives: Plasmodium falciparum cerebral malaria remains a major health problemin India. The efficacy of treatment of cerebral malaria lies in its early diagnosis through rapid diagnosticmethods. ParaSights-F test detects HRP-2 antigen secreted by parasitised red blood cells andquantitative buffy coat assay (QBC is examination of buffy coat for the presence of malarial parasitestained with acridine orange. This study was performed to evaluate the effectiveness of ParaSight-F test and QBC assay as diagnostic methods in the patients of cerebral malaria.Methods: Fifty clinically diagnosed patients of cerebral malaria were included in the study.ParaSight-F test, QBC and conventional blood smear examination was done. Patients who were incoma and there were no obvious features of bacterial or viral etiology were investigated for cerebralmalaria by these diagnostic methods.Results: ParaSight-F test, QBC and peripheral blood smears were examined. Patients were followedupfor signs of clinical recovery. ParaSight-F test was positive in 47 patients, QBC in 46 while bloodsmear examination was positive in 28 cases.Interpretation & conclusion: Sensitivity and specificity of ParaSight-F test were found to be 96.6 and94% while QBC showed 97.8 and 100% respectively. ParaSight-F test and QBC were found to be novelmethods for diagnosis of cerebral malaria especially in the cases where diagnosis can not be made byconventional blood smear examination due to low parasitaemia. These rapid diagnostic methods helpin early therapeutic intervention.

  20. Screening of Toxin Mutant of Dickeya zeae and Its Biological Characters

    Institute of Scientific and Technical Information of China (English)

    Jingyi ZHANG; Yutao WANG; Yanchang LI; Qiongguang LIU

    2013-01-01

    [Objective] This study aimed to screen toxin mutant of Dickeya zeae (Er-winia chrysanthemi pv. zeae) and investigate its biological characters. [Method] We obtained a toxin mutant strain D. zeae Ech7-3-42 by using acridine orange as a mutagenic agent and compared their biological characteristics and virulence between the toxin mutant and wild strain. [Result] There was no significant difference in pectin lyase, protease, cellulase and the production of extracellular polysaccharide and lipopolysaccharide, but significant difference in toxin biological activities and vir-ulence. Ech7-3-42 mutant did not produce toxin, as wel as the loss of virulence on rice and HR on tobacco, but did not lose the ability to soft rot on potato. Mutant strain Ech7-3-42 can infect rice root and then enriched in the root neck and stalk, but it could not cause rice foot rot. Dickeya zeae (wild and mutant strain) could be detected by PCR in the root neck and below the 1-2 cm long stem area, but could not be detected in the leaves. [Conclusion] We believed that toxin may be one of the important factors for D. zeae virulence on rice.

  1. Tri-modal confocal mosaics detect residual invasive squamous cell carcinoma in Mohs surgical excisions

    Science.gov (United States)

    Gareau, Dan; Bar, Anna; Snaveley, Nicholas; Lee, Ken; Chen, Nathaniel; Swanson, Neil; Simpson, Eric; Jacques, Steve

    2012-06-01

    For rapid, intra-operative pathological margin assessment to guide staged cancer excisions, multimodal confocal mosaic scan image wide surgical margins (approximately 1 cm) with sub-cellular resolution and mimic the appearance of conventional hematoxylin and eosin histopathology (H&E). The goal of this work is to combine three confocal imaging modes: acridine orange fluorescence (AO) for labeling nuclei, eosin fluorescence (Eo) for labeling cytoplasm, and endogenous reflectance (R) for marking collagen and keratin. Absorption contrast is achieved by alternating the excitation wavelength: 488 nm (AO fluorescence) and 532 nm (Eo fluorescence). Superposition and false-coloring of these modes mimics H&E, enabling detection of cutaneous squamous cell carcinomas (SCC). The sum of mosaic Eo+R is false-colored pink to mimic the appearance of eosin, while the AO mosaic is false-colored purple to mimic the appearance of hematoxylin in H&E. In this study, mosaics of 10 Mohs surgical excisions containing invasive SCC, and five containing only normal tissue were subdivided for digital presentation equivalent to 4× histology. Of the total 50 SCC and 25 normal sub-mosaics presented, two reviewers made two and three type-2 errors (false positives), respectively. Limitations to precisely mimic H&E included occasional elastin staining by AO. These results suggest that confocal mosaics may effectively guide staged SCC excisions in skin and other tissues.

  2. In vivo antitumor activity of biosynthesized silver nanoparticles using Ficus religiosa as a nanofactory in DAL induced mice model.

    Science.gov (United States)

    Antony, Jacob Joe; Sithika, Mohamed Ali Ayisha; Joseph, Thomas Amal; Suriyakalaa, Udhayaraj; Sankarganesh, Arunachalam; Siva, Durairaj; Kalaiselvi, Seenivasan; Achiraman, Shanmugam

    2013-08-01

    Ficus religiosa leaf extract was chosen as a reducing agent to fabricate silver nanoparticles (AgNPs) by a simple, cost-effective and eco-friendly process with the aim of treating Dalton's ascites lymphoma (DAL) in mice model. The formation of synthesized nanoparticles were characterized by UV-visible analysis (UV-vis), Fourier transform infra-red (FT-IR), transmission electron microscopy (TEM), X-ray diffraction (XRD) and zeta potential analyses. A peak at 431nm indicated the surface plasmon resonance of AgNPs. FTIR studies indicated polyphenols and proteins as possible encapsulates. TEM analysis showed particles size in the range of 5-35nm. Healthy Swiss Albino mice (30-35g) were intraperitoneally induced with DAL cells and treated with F. religiosa derived AgNPs at a dose of 50μg/ml. Blood and liver tissues were collected subsequent to dissection and subjected to hematological, biochemical and anticancer assays. Hematological and biochemical analyses revealed revival after treating with F. religiosa derived AgNPs. Antioxidant activity results further proved supportive evidence. The apoptosis inducing effect of AgNPs was observed through acridine orange staining (AO and EB) and DNA fragmentation assay. Anti- angiogenic activity was confirmed by observing vessel development. All these observations indicate that the AgNPs were effective in treatment of DAL. PMID:23537836

  3. An evaluation on the adherence of Candida albicans to different denture- base materials

    Directory of Open Access Journals (Sweden)

    Savabi O

    2004-02-01

    Full Text Available The surface topography of denture base material is an important factor for the"nadhesion of Candida albicans and other microorganisms."nPurpose: The aim of this study was to evaluate the adherence of Candida albicans to four types of denture"nbase materials (Acropars acrylic resin, Meliodent acrylic resin, rough and smooth surfaces of Molloplast B."nMaterials and Methods: Seven blocks of two types of acrylic resins and ten blocks of silicone with one"nrough and one smooth surface were made and incubated in a suspension of Candida albicans. After washing,"nthe blocks were stained with acridine orange and examined under fluorescent microscope. For statistical"nanalysis ANOVA and Duncan tests were used."nResults: It was observed that Candida adhesion to rough surfaces of acrylic resins and silicone was"nsignificantly more than polished surfaces of acrylic resins and smooth silicone (PO.0001. However, no"nstatistical significant difference was found between polished acrylic resins surfaces and smooth silicone."nConclusion: Significant differences in the adherence of Candida to the surfaces of different denture base"nmaterials are due to differences in surface topography, chemical, physical and hydrophobic properties so it is"nrecommended to minimize the roughness and irregularities of denture base.

  4. Experimental studies on islets isolation, purification and function in rats.

    Science.gov (United States)

    Pang, Xinlu; Xue, Wujun; Feng, Xinshun; Tian, Xiaohui; Teng, Yan; Ding, Xiaoming; Pan, Xiaoming; Guo, Qi; He, Xiaoli

    2015-01-01

    To develop a simple and effective method of islet isolation and purification in rats. Collagenase P was injected into pancreatic duct followed by incubation in water bath to digest the pancreas and isolate islet, then discontinuous gravity gradient purification was used to purify the islet. The purified islets were identified by dithizone staining. The viability of islets was assessed by fluorescence staining of acridine orange (AO) and propidium iodide (PI). The function of purified islets was determined by glucose-stimulated insulin release test and transplantation of rat with streptozocin-induced diabetes. 738±193 islets were recovered after purification. The average purity was 77±13%, the viability of islets was more than 95%. When inspected by glucose stimulation, the secreted insulin concentration was 24.31±5.47 mIU/L when stimulated by low concentration glucose and 37.62±4.29 mIU/L by high concentration glucose. There was significant difference between the two phases (Pmethod of injecting collagenase into pancreatic duct followed by incubation in water bath and purification using discontinuous gravity gradient. PMID:26885021

  5. Self-bioremediation of cork-processing wastewaters by (chloro)phenol-degrading bacteria immobilised onto residual cork particles.

    Science.gov (United States)

    del Castillo, I; Hernández, P; Lafuente, A; Rodríguez-Llorente, I D; Caviedes, M A; Pajuelo, E

    2012-04-15

    Cork manufacturing is a traditional industry in Southern Europe, being the main application of this natural product in wine stoppers and insulation. Cork processing begins at boiling the raw material. As a consequence, great volumes of dark wastewaters, with elevated concentrations of chlorophenols, are generated, which must be depurated through costly physicochemical procedures before discarding them into public water courses. This work explores the potential of bacteria, isolated from cork-boiling waters storage ponds, in bioremediation of the same effluent. The bacterial population present in cork-processing wastewaters was analysed by DGGE; low bacterial biodiversity was found. Aerobic bacteria were isolated and investigated for their tolerance against phenol and two chlorophenols. The most tolerant strains were identified by sequencing 16S rDNA. The phenol-degrading capacity was investigated by determining enzyme activities of the phenol-degrading pathway. Moreover, the capacity to form biofilms was analysed in a microtitre plate assay. Finally, the capacity to form biofilms onto the surface of residual small cork particles was evaluated by acridine staining followed by epifluorescence microscopy and by SEM. A low-cost bioremediation system, using phenol-degrading bacteria immobilised onto residual cork particles (a by-product of the industry) is proposed for the remediation of this industrial effluent (self-bioremediation).

  6. Induction of allogeneic unresponsiveness in adult dogs by irradiation and bone marrow transplantation: Implication of Ia-positive bone marrow stem cells

    International Nuclear Information System (INIS)

    A number of investigators have suggested in recent years that placement of hemopoietic cells into an irradiated host milieu may trigger such cells to undergo a transient cycle of replication and differentiation which recapitulates the events of immunological ontogeny-including fetal erythropoiesis, production of newborn Υ-chains in adults, and generation of fetal and newborn-type lymphoid cells. In an extension of this hypothesis to transplantation, supralethally irradiated dogs were reconstituted with their own stored marrow, followed within 12 to 18 hr by the transplanation of a kidney allograft obtained from a DLA identical donor. This sequence resulted in long-term unresponsiveness to the transplanted kidneys in the recipients without further treatment. The 60% incidence of success obtained with this procedure could be improved further if the host's own stored marrow was treated in vitro with methylprednisolone (MPd) prior to replacement of the marrow following irradiation. In an attempt to analyze the possible changes in the cellular composition of marrow that might have been associated with this result, a serial cytofluorographic analysis of marrow before and after treatment with MPd was performed. For this purpose, cell samples obtained before and after exposure of bone marrow to MPd were studied in an Ortho 50H Cell Sorter after staining with acridine orange by using green fluorescence for DNA and red fluroescence for RNA, or, alternatively, using 900 scatter for the X axis and a narrow forward scatter for the Y axis, without addition of any stain

  7. Binding of anti-prion agents to glycosaminoglycans: Evidence from electronic absorption and circular dichroism spectroscopy

    International Nuclear Information System (INIS)

    The polyanionic glycosaminoglycans (GAGs) are intimately involved in the pathogenesis of protein conformational disorders such as amyloidosis and prion diseases. Several cationic agents are known to exhibit anti-prion activity but their mechanism of action is poorly understood. In this study, UV absorption and circular dichroism (CD) spectroscopic techniques were used to investigate the interaction between heparin and chondroitin-6-sulfate and anti-prion drugs including acridine, quinoline, and phenothiazine derivatives. UV band hypochromism of (±)-quinacrine, (±)-primaquine, tacrine, quinidine, chlorpromazine, and induced CD spectra of (±)-quinacrine upon addition of GAGs provided evidence for the GAG binding of these compounds. The association constants (∼106-107 M-1) estimated from the UV titration curves show high-affinity drug-heparin interactions. Ionic strength-dependence of the absorption spectra suggested that the interaction between GAGs and the cationic drugs is principally electrostatic in nature. Drug binding differences of heparin and chondroitin-6-sulfate were attributed to their different negative charge density. These results call the attention to the alteration of GAG-prion/GAG-amyloid interactions by which these compounds might exert their anti-prion/anti-amyloidogenic activities

  8. Host–guest composite materials of dyes loaded zeolite LTL for antenna applications

    Energy Technology Data Exchange (ETDEWEB)

    Insuwan, W. [Rajamangala University of Technology Isan Surin Campus, Facculty of Agriculture and Technology, Surin 32000 (Thailand); Jungsuttiwong, S. [Center for Organic Electronic and Alternative Energy, Department of Chemistry and Center of Excellence for Innovation in Chemistry, Faculty of Science, Ubon Ratchathani University, Ubon Ratchathani 34190 (Thailand); Rangsriwatananon, K., E-mail: kunwadee@sut.ac.th [School of Chemistry, Institute of Science, Suranaree University of Technology, Nakhon Ratchasima 30000 (Thailand)

    2015-05-15

    This research work directly focuses on a new feasible light harvesting antenna material constructed with Acridine hydrochloride (Ac)/Acriflavine hydrochloride (AF), as donor/acceptor for energy transfer, loaded on a round shape zeolite LTL (K-LTL and H-LTL). The energy transfer was monitored by absorption and fluorescence spectra while the calculated Förster distance (R{sub DA}) and Quenching efficiency (%Q) of Ac/AF on K-LTL and H-LTL varied between 22.0 Å to 19.6 Å and 71.4% to 65.5%, respectively. Also, it was found that the microenvironment of a solid host such as K-LTL and H-LTL has significantly influenced the fluorescence spectra of Ac/AF on H-LTL approximately 50 nm longer than that on K-LTL. - Highlights: • New antenna materials have been performed using dyes loaded on zeolite LTL. • Light emission takes place from acriflavine hydrochloride (AF) due to fluorescence resonance energy transfer (FRET). • The microenvironment of zeolite LTL has significantly influenced the fluorescence spectra.

  9. Spectroscopic study of fast-neutron-irradiated chromatin

    Energy Technology Data Exchange (ETDEWEB)

    Radu, L. [V. Babes National Inst., Dept. of Molecular Genetics, Bucharest (Romania)]. E-mail: serbanradu@pcnet.ro; Gazdaru, D. [Bucharest Univ., Dept. of Biophysics, Physics Faculty, Bucharest (Romania); Constantinescu, B. [H. Hulubei National Inst., Dept. of Cyclotron, Bucharest (Romania)

    2004-02-01

    The effects produced by fast neutrons (0-100 Gy) on chromatin structure were analyzed by (i) [{sup 1}H]-NMR spectroscopy, (ii) time resolved spectroscopy, and (iii) fluorescence resonance energy transfer (FRET). Two types of chromatin were tested: (i) a chromatin from a normal tissue (liver of Wistar rats) and (ii) a chromatin from a tumoral tissue (Guerin limphotrope epithelioma, a rat solid tumor). The fast-neutron action on chromatin determines greater values of the [{sup 1}H]-NMR transverse relaxation time, indicating a more injured structure. Time-resolved fluorescence measurements show that the relative contribution of the excited state lifetime of bound ethidium bromide to chromatin DNA diminishes with increasing irradiation doses. This reflects the damage that occurs in DNA structure: production of single- and double-strand breaks due to sugar and base modifications. By the FRET method, the distance between dansyl chloride and acridine orange coupled at chromatin was determined. This distance increases upon fast-neutron action. The radiosensitivity of the tumor tissue chromatin seems higher than that of the normal tissue chromatin, probably because of its higher (loose) euchromatin/(compact) heterochromatin ratio. As the values of the physical parameters analyzed are specific for a determined dose, the establishment of these parameters may constitute a criterion for the microdosimetry of chromatin radiolesions produced by fast neutrons. (author)

  10. Effects of fast neutrons on chromatin: dependence on chromatin structure

    Energy Technology Data Exchange (ETDEWEB)

    Radu, L. [Dept. of Molecular Genetics, V. Babes National Inst., Bd. Timisoara, Bucharest (Romania); Constantinescu, B. [Dept. of Cyclotron, H. Hulubei National Inst., Bucharest (Romania); Gazdaru, D. [Dept. of Biophysics, Physics Faculty, Univ. of Bucharest (Romania)

    2002-07-01

    The effects of fast neutrons (10-100 Gy) on chromatin extracted from normal (liver of Wistar rats) and tumor (Walker carcinosarcoma maintained on Wistar rats) tissues were compared. The spectroscopic assays used were (i) chromatin intrinsic fluorescence, (ii) time-resolved fluorescence of chromatin-proflavine complexes, and (iii) fluorescence resonance energy transfer (FRET) between dansyl chloride and acridine orange coupled to chromatin. For both normal and tumor chromatin, the intensity of intrinsic fluorescence specific for acidic and basic proteins decreased with increasing dose. The relative contributions of the excited-state lifetime of proflavine bound to chromatin were reduced upon fast-neutron irradiation, indicating a decrease in the proportion of chromatin DNA available for ligand binding. The Forster energy transfer efficiencies were also modified by irradiation. These effects were larger for chromatin from tumor tissue. In the range 0-100 Gy, fast neutrons induced alterations in DNA and acidic and basic proteins, as well as in global chromatin structure. The radiosensitivity of chromatin extracted from tumor tissue seems to be higher than that of chromatin extracted from normal tissue, probably because of its higher euchromatin (loose)-heterochromatin (compact) ratio. (author)

  11. Self-bioremediation of cork-processing wastewaters by (chloro)phenol-degrading bacteria immobilised onto residual cork particles.

    Science.gov (United States)

    del Castillo, I; Hernández, P; Lafuente, A; Rodríguez-Llorente, I D; Caviedes, M A; Pajuelo, E

    2012-04-15

    Cork manufacturing is a traditional industry in Southern Europe, being the main application of this natural product in wine stoppers and insulation. Cork processing begins at boiling the raw material. As a consequence, great volumes of dark wastewaters, with elevated concentrations of chlorophenols, are generated, which must be depurated through costly physicochemical procedures before discarding them into public water courses. This work explores the potential of bacteria, isolated from cork-boiling waters storage ponds, in bioremediation of the same effluent. The bacterial population present in cork-processing wastewaters was analysed by DGGE; low bacterial biodiversity was found. Aerobic bacteria were isolated and investigated for their tolerance against phenol and two chlorophenols. The most tolerant strains were identified by sequencing 16S rDNA. The phenol-degrading capacity was investigated by determining enzyme activities of the phenol-degrading pathway. Moreover, the capacity to form biofilms was analysed in a microtitre plate assay. Finally, the capacity to form biofilms onto the surface of residual small cork particles was evaluated by acridine staining followed by epifluorescence microscopy and by SEM. A low-cost bioremediation system, using phenol-degrading bacteria immobilised onto residual cork particles (a by-product of the industry) is proposed for the remediation of this industrial effluent (self-bioremediation). PMID:22265252

  12. Structured superparamagnetic nanoparticles for high performance mediator of magnetic fluid hyperthermia: synthesis, colloidal stability and biocompatibility evaluation.

    Science.gov (United States)

    Thorat, N D; Otari, S V; Bohara, R A; Yadav, H M; Khot, V M; Salunkhe, A B; Phadatare, M R; Prasad, A I; Ningthoujam, R S; Pawar, S H

    2014-09-01

    Core-shell structures with magnetic core and metal/polymer shell provide a new opportunity for constructing highly efficient mediator for magnetic fluid hyperthermia. Herein, a facile method is described for the synthesis of superparamagnetic LSMO@Pluronic F127 core-shell nanoparticles. Initially, the surface of the LSMO nanoparticles is functionalized with oleic acid and the polymeric shell formation is achieved through hydrophobic interactions with oleic acid. Each step is optimized to get good dispersion and less aggregation. This methodology results into core-shell formation, of average diameter less than 40 nm, which was stable under physiological conditions. After making a core-shell formulation, a significant increase of specific absorption rate (up to 300%) has been achieved with variation of the magnetization (Fe3O4. MTT assay is used to evaluate the toxicity of bare and core-shell MNPs. The mechanism of cell death by necrosis and apoptosis is studied with sequential staining of acridine orange and ethidium bromide using fluorescence and confocal microscopy. The present work reports a facile method for the synthesis of core-shell structure which significantly improves SAR and biocompatibility of bare LSMO MNPs, indicating potential application for hyperthermia. PMID:25063164

  13. DNA binding and anti-cancer activity of redox-active heteroleptic piano-stool Ru(II), Rh(III), and Ir(III) complexes containing 4-(2-methoxypyridyl)phenyldipyrromethene.

    Science.gov (United States)

    Gupta, Rakesh Kumar; Pandey, Rampal; Sharma, Gunjan; Prasad, Ritika; Koch, Biplob; Srikrishna, Saripella; Li, Pei-Zhou; Xu, Qiang; Pandey, Daya Shankar

    2013-04-01

    The synthesis of four novel heteroleptic dipyrrinato complexes [(η(6)-arene)RuCl(2-pcdpm)] (η(6)-arene = C6H6, 1; C10H14, 2) and [(η(5)-C5Me5)MCl(2-pcdpm)] (M = Rh, 3; Ir, 4) containing a new chelating ligand 4-(2-methoxypyridyl)-phenyldipyrromethene (2-pcdpm) have been described. The complexes 1-4 have been fully characterized by various physicochemical techniques, namely, elemental analyses, spectral (ESI-MS, IR, (1)H, (13)C NMR, UV/vis) and electrochemical studies (cyclic voltammetry (CV) and differential pulse voltammetry (DPV)). Structures of 3 and 4 have been determined crystallographically. In vitro antiproliferative and cytotoxic activity of these complexes has been evaluated by trypan blue exclusion assay, cell morphology, apoptosis, acridine orange/ethidium bromide (AO/EtBr) fluorescence staining, and DNA fragmentation assay in Dalton lymphoma (DL) cell lines. Interaction of 1-4 with calf thymus DNA (CT DNA) has also been supported by absorption titration and electrochemical studies. Our results suggest that in vitro antitumor activity of 1-4 lies in the order 2 > 1 > 4 > 3. PMID:23477351

  14. Synthesis and antitumor activity of conjugates of muramyldipeptide or normuramyldipeptide with hydroxyacridine/acridone derivatives.

    Science.gov (United States)

    Dzierzbicka, Krystyna; Kołodziejczyk, Aleksander M

    2003-01-01

    A series of MDP (muramyldipeptide) or nor-MDP (normuramyldipeptide) analogues modified at the C-terminus post of the molecule by a formation of an ester bond between the carboxylic group of isoglutamine and the hydroxyl function of the respective derivatives of 4-carboxamide-acridine/9-acridone or 1-nitro-9-hydroxyalkylaminoacridines were synthesized as potential anticancer agents. The compounds O-(1-O-benzyl-N-acetyl-muramyl-l-alanyl-d-gamma-isoglutaminyl)-9-(ethylamino)-1-nitroacridine ester 3j and O-(1-O-benzyl-N-acetyl-muramyl-l-alanyl-d-gamma-isoglutaminyl)-9-propylamino-1-nitroacridine ester 3k exhibited high in vitro cytotoxic activity against a panel of human cell lines, prostate cancer and AIDS-related lymphoma (ARL). Analogue 3j was also active in vivo in the hollow fiber assay. Antitumor activity of both compounds were tested in vivo against difference human tumor xenograft, but only analogue 3k showed in vivo activity against sc UACC-62 melanoma in mice.

  15. Patterned macroarray plates in comparison of bacterial adhesion inhibition of tantalum, titanium, and chromium compared with diamond-like carbon.

    Science.gov (United States)

    Levon, Jaakko; Myllymaa, Katja; Kouri, Vesa-Petteri; Rautemaa, Riina; Kinnari, Teemu; Myllymaa, Sami; Konttinen, Yrjö T; Lappalainen, Reijo

    2010-03-15

    Staphylococcus aureus device-related infection is a common complication in implantology. Bacterial adhesion on implant surfaces is the initial step in the infective process. The aim was to develop a method suitable for quantitative bacterial adherence studies and to test a new diamond-like carbon (DLC) coating against commonly used metallic biomaterials with regards to Staphylococcus aureus adhesion. Patterned silicon chips with spots of tantalum, titanium, chromium, and DLC were produced using ultraviolet lithography and physical vapor deposition. These patterned chips were used as such or glued to array plates, pretreated with serum and exposed to S. aureus (S-15981) for 90 min, followed by acridine orange staining and fluorescence microscopy. An adhesion index showed that the ranking order of the biomaterials was titanium, tantalum, chromium, and DLC, with the DLC being clearly most resistant against colonization with S. aureus. Micropatterned surfaces are useful for quantitative comparison of bacterial adherence on different biomaterials. In the presence of serum, DLC is superior in its ability to resist adhesion and colonization by S. aureus compared with the commonly used biomaterial metals tantalum, titanium, and chromium. PMID:19437436

  16. Plasmid-mediated biodegradation of the anionic surfactant sodium dodecyl sulphate, by Pseudomonas aeruginosa S7.

    Science.gov (United States)

    Yeldho, Deepthi; Rebello, Sharrel; Jisha, M S

    2011-01-01

    Sodium dodecyl sulphate (SDS), an anionic surfactant, has been used extensively due to its low cost and excellent foaming properties. Fifteen different bacterial isolates capable of degrading SDS were isolated from detergent contaminated soil by enrichment culture technique and the degradation efficiency was assessed by Methylene Blue Active Substances (MBAS) assay. The most efficient SDS degrading isolate was selected and identified as Pseudomonas aeruginosa S7. The selected isolate was found to harbor a single 6-kb plasmid. Acridine orange, ethidium bromide, SDS and elevated temperatures of incubation failed to cure the plasmid. The cured derivatives of SDS degrading Pseudomonas aeruginosa were obtained only when ethidium bromide and elevated temperature (40 °C) were used together. Transformation of E. coli DH5α with plasmid isolated from S7 resulted in subsequent growth of the transformants on minimal salt media with SDS (0.1%) as the sole source of carbon. The SDS degradation ability of S7 and the transformant was found to be similar as assessed by Methylene Blue Active Substance Assay. The antibiotic resistance profiles of S7, competent DH5α and transformant were analyzed and it was noted that the transfer of antibiotic resistance correlated with the transfer of plasmid as well as SDS degrading property.

  17. Mercury-Pollution Induction of Intracellular Lipid Accumulation and Lysosomal Compartment Amplification in the Benthic Foraminifer Ammonia parkinsoniana.

    Science.gov (United States)

    Frontalini, Fabrizio; Curzi, Davide; Cesarini, Erica; Canonico, Barbara; Giordano, Francesco M; De Matteis, Rita; Bernhard, Joan M; Pieretti, Nadia; Gu, Baohua; Eskelsen, Jeremy R; Jubb, Aaron M; Zhao, Linduo; Pierce, Eric M; Gobbi, Pietro; Papa, Stefano; Coccioni, Rodolfo

    2016-01-01

    Heavy metals such as mercury (Hg) pose a significant health hazard through bioaccumulation and biomagnification. By penetrating cell membranes, heavy metal ions may lead to pathological conditions. Here we examined the responses of Ammonia parkinsoniana, a benthic foraminiferan, to different concentrations of Hg in the artificial sea water. Confocal images of untreated and treated specimens using fluorescent probes (Nile Red and Acridine Orange) provided an opportunity for visualizing the intracellular lipid accumulation and acidic compartment regulation. With increased Hg over time, we observed an increased number of lipid droplets, which may have acted as a detoxifying organelle where Hg is sequestered and biologically inactivated. Further, Hg seems to promote the proliferation of lysosomes both in terms of number and dimension that, at the highest level of Hg, resulted in cell death. We report, for the first time, the presence of Hg within the foraminiferal cell: at the basal part of pores, in the organic linings of the foramen/septa, and as cytoplasmic accumulations. PMID:27603511

  18. Experimental Study of Rat Beta Islet Cells Cultured under Simulated Microgravity Conditions

    Institute of Scientific and Technical Information of China (English)

    ChunSONG; Xiu-QingDUAN; XiLI; Li-OuHAN; PingXU; Chun-FangSONG:; Lian-HongJIN

    2004-01-01

    To observe the effects of simulated microgravity on beta islet cell culture, we have compared the survival rates and the insulin levels of the isolated rat islet cells cultured at micro- and normal gravity conditions. The survival rates of the cells cultured were determined by acridine orange-propidium iodide double-staining on day 3,7 and 14. The morphology of the cells was observed by electron microscopy.Insulin levels were measured by radio immuno assays. Our results show that the cell number cultured underthe microgravity condition is significantly higher than that under the routine condition (P<0.01). Some tubular structure shown by transmission electron microscopy, possibly for the transport of nutrients, were formed intercellularly in the microgravity cultured group on day 7. There were also abundant secretion particles and mitochondria in the cytoplasm of the cells. Scanning electron microscopy showed that there were holes formed between each islet, possibly connecting with the nutrient transport tubules. The microgravity cultured group also has higher insulin levels in the media as compared with the control group (P<0.01). Our results indicate that microgravity cultivation of islet cells has advantages over the routine culture methods.

  19. Inositol Hexakisphosphate Mediates Apoptosis in Human Breast Adenocarcinoma MCF-7 Cell Line via Intrinsic Pathway

    Science.gov (United States)

    Agarwal, Rakhee; Ali, Nawab

    2010-04-01

    Inositol polyphosphates (InsPs) are naturally occurring compounds ubiquitously present in plants and animals. Inositol hexakisphosphate (InsP6) is the most abundant among all InsPs and constitutes the major portion of dietary fiber in most cereals, legumes and nuts. Certain derivatives of InsPs also regulate cellular signaling mechanisms. InsPs have also been shown to reduce tumor formation and induce apoptosis in cancerous cells. Therefore, in this study, the effects of InsPs on apoptosis were studied in an attempt to investigate their potential anti-cancer therapeutic application and understand their mechanism of action. Acridine orange and ethidium bromide staining suggested that InsP6 dose dependently induced apoptosis in human breast adenocarcinoma MCF-7 cells. Among InsPs tested (InsP3, InsP4, InsP5, and InsP6), InsP6 was found to be the most effective in inducing apoptosis. Furthermore, effects of InsP6 were found most potent inducing apoptosis. Etoposide, the drug known to induce apoptosis in both in vivo and in vitro, was used as a positive control. Western blotting experiments using specific antibodies against known apoptotic markers suggested that InsP6 induced apoptotic changes were mediated via an intrinsic apoptotic pathway.

  20. The novel pterostilbene derivative ANK-199 induces autophagic cell death through regulating PI3 kinase class III/beclin 1/Atg‑related proteins in cisplatin‑resistant CAR human oral cancer cells.

    Science.gov (United States)

    Hsieh, Min-Tsang; Chen, Hao-Ping; Lu, Chi-Cheng; Chiang, Jo-Hua; Wu, Tian-Shung; Kuo, Daih-Huang; Huang, Li-Jiau; Kuo, Sheng-Chu; Yang, Jai-Sing

    2014-08-01

    Pterostilbene is an effective chemopreventive agent against multiple types of cancer cells. A novel pterostilbene derivative, ANK-199, was designed and synthesized by our group. Its antitumor activity and mechanism in cisplatin-resistant CAR human oral cancer cells were investigated in this study. Our results show that ANK-199 has an extremely low toxicity in normal oral cell lines. The formation of autophagic vacuoles and acidic vesicular organelles (AVOs) was observed in the ANK-199-treated CAR cells by monodansylcadaverine (MDC) and acridine orange (AO) staining, suggesting that ANK-199 is able to induce autophagic cell death in CAR cells. Neither DNA fragmentation nor DNA condensation was observed, which means that ANK-199-induced cell death is not triggered by apoptosis. In accordance with morphological observation, 3-MA, a specific inhibitor of PI3K kinase class III, can inhibit the autophagic vesicle formation induced by ANK-199. In addition, ANK-199 is also able to enhance the protein levels of autophagic proteins, Atg complex, beclin 1, PI3K class III and LC3-II, and mRNA expression of autophagic genes Atg7, Atg12, beclin 1 and LC3-II in the ANK-199-treated CAR cells. A molecular signaling pathway induced by ANK-199 was therefore summarized. Results presented in this study show that ANK-199 may become a novel therapeutic reagent for the treatment of oral cancer in the near future (patent pending).

  1. Electroluminescence of organic light-emitting diodes with an ultra-thin layer of dopant

    Energy Technology Data Exchange (ETDEWEB)

    Li Weizhi [State Key Lab of Electronic Thin Films and Integrated Devices, School of Optoelectronic Information, University of Electronic Science and Technology of China (UESTC), Chengdu 610054 (China); Yu Junsheng [State Key Lab of Electronic Thin Films and Integrated Devices, School of Optoelectronic Information, University of Electronic Science and Technology of China (UESTC), Chengdu 610054 (China)], E-mail: jsyu@uestc.edu.cn; Wang, Tao [State Key Lab of Electronic Thin Films and Integrated Devices, School of Optoelectronic Information, University of Electronic Science and Technology of China (UESTC), Chengdu 610054 (China); Jiang, Yadong [State Key Lab of Electronic Thin Films and Integrated Devices, School of Optoelectronic Information, University of Electronic Science and Technology of China (UESTC), Chengdu 610054 (China)], E-mail: jiangyd@uestc.edu.cn; Wei, Bangxiong [State Key Lab of Electronic Thin Films and Integrated Devices, School of Optoelectronic Information, University of Electronic Science and Technology of China (UESTC), Chengdu 610054 (China)

    2008-03-15

    Conventional fluorescent dyes, i.e., 4-(dicyanomethylene)-2-t-butyl-6(1,1,7,7-tetramethyljulolidyl-9-enyl)-4H-pyran (DCJTB), 5,12-dihydro-5,12-dimethylquino [2,3-b]acridine-7,14-dione (DMQA) and 5,6,11,12-tetraphenylnaphthacene (Rubrene), were used to investigate the performance of organic light-emitting diodes (OLEDs) based on indium tin oxide (ITO)/N,N'-bis-(1-naphthyl)-N,N'-diphenyl-1,1'-biphenyl-4,4'-diamine (NPB)/tris-(8-hydroxyquinolate)-aluminum (Alq{sub 3})/MgAg. The dyes were either inserted into devices as an ultra-thin film at the NPB/Alq{sub 3} interface by sequential evaporation, or doped into the Alq{sub 3} emission layer by co-evaporation with the doping ratio about 2%. Electroluminescence (EL) spectra of devices indicated that concentration quenching effect (CQE) of the dye-dopant was slightly bigger in the former than in the latter, while the degrees of CQE for three dopants are in the order of DMQA > DCJTB > Rubrene suggested by the difference in EL spectra and performances of devices. In addition, EL process of device with an ultra-thin layer of dopant is dominated by direct carrier trapping (DCT) process due to almost no holes recombine with electrons in Alq{sub 3}-host layer.

  2. Study on electroluminescence processes in dye-doped organic light-emitting diodes

    Energy Technology Data Exchange (ETDEWEB)

    Li Weizhi [State Key Laboratory of Electronic Thin Films and Integrated devices, School of Optoelectronic Information, University of Electronic Science and Technology of China (UESTC), Chengdu 610054 (China)], E-mail: leewz@uestc.edu.cn; Jiang Yadong; Wang Tao [State Key Laboratory of Electronic Thin Films and Integrated devices, School of Optoelectronic Information, University of Electronic Science and Technology of China (UESTC), Chengdu 610054 (China)

    2008-07-15

    Electroluminescence (EL) mechanism of dye-doped organic light-emitting diodes (OLEDs) was investigated by using three familiar fluorescent dyes, i.e., 5,12-Dihydro-5,12-dimethylquino [2,3-b]acridine-7,14-dione (DMQA), 4-(dicyanomethylene)-2-t-butyl-6(1,1,7,7-tetramethyljulolidyl-9-enyl) -4H-pyran(DCJTB), and 5,6,11,12-tetraphenylnaphthacene (Rubrene). EL spectra of the doped devices with structure of indium tin oxide (ITO)/N,N'-bis-(1-naphthyl)-N,N'-diphenyl-1,1'-biphenyl-4,4'- diamine (NPB) (40 nm)/tris-(8-hydroxyquinolate)-aluminum (Alq{sub 3}) (x nm, x=0-40 nm)/dye: Alq{sub 3} (weight ratio{approx}1%, 2 nm)/Alq{sub 3} (48-x nm)/MgAg indicated that direct carrier trapping (DCT) process dominated light emission of devices. As a result, investigation of carrier-recombination site via doping, which is conventionally applied in OLEDs, is questionable since the doping site and the dopant itself may significantly influence the carrier-recombination process in the doped devices.

  3. Design, synthesis and biological evaluation of Arylpiperazine-based novel Phthalimides: Active inducers of testicular germ cell apoptosis

    Indian Academy of Sciences (India)

    ANIL K SINGH; JITENDER K BHARDWAJ; ANA OLIVAL; YOGESH KUMAR; AVIJIT PODDER; ANKUR MAHESHWARI; RENUKA AGRAWAL; N LATHA; BRAJENDRA K SINGH; HELENA TOMÁS; JOÃO RODRIGUES; RAM KISHAN; B RUPINI; BRIJESH RATHI

    2016-08-01

    Understanding of apoptosis or programmed cell death has provided the basis for novel therapeutics that has resulted in rationally designed anticancer strategies. Recently, inducers of apoptosis have been used in cancer therapy. In this work, we describe the role of chiral phthalimides functionalized with piperazines aspotential apoptotic inducers. The listed twenty phthalimides were assessed for their in vitro apoptotic activity against testicular germ cells. All phthalimides showed a significant apoptotic response (∼39 to ∼68%). TUNEL assay and acridine orange fluorescence staining were carried out to investigate the molecular mechanismsresponsible for the cell death. Phthalimides exhibited substantial apoptotic induction following the intrinsic pathway mechanism. Studies advocated that the apoptotic induction was mediated through caspase-9, caspase-3, JNK MAP kinase and tumor suppressor p53, which was accompanied by DNA fragmentation and nuclearcondensation. Besides, the best five phthalimides regarding apoptotic action were evaluated for in vitro cytotoxic effects against CAL-72 and MCF-7 cancer cell lines. Compounds showed efficient killing of cancer cells. This discovery of functionalized phthalimides as apoptotic inducers would be highly valuable in understanding the mechanism of apoptosis at the molecular level and opens up new possibilities for therapeutic strategies.

  4. A cytotoxic meroterpenoid benzoquinone from roots of Cordia globosa.

    Science.gov (United States)

    Alencar de Menezes, Jane Eire; Lemos, Telma Leda; Pessoa, Otília Deusdênia; Braz-Filho, Raimundo; Montenegro, Raquel C; Wilke, Diego Veras; Costa-Lotufo, Letícia V; Pessoa, Cláudia; de Moraes, Manoel Odorico; Silveira, Edilberto R

    2005-01-01

    (1a S*,1b S*,7a S*,8a S*)-4,5-Dimethoxy-1a,7a-dimethyl-1,1a,1b,2,7, 7a,8,8a-octahydrocyclopropa cyclopenta[1,2-b]naphthalene-3,6-dione (1), a new meroterpenoid benzoquinone, and microphyllaquinone (2), a known naphthoquinone, have been isolated from roots of Cordia globosa. Both structure determinations were performed by conventional spectroscopic methods, including inverse detection NMR techniques, and by comparison with data from the literature for related compounds. Compound 1 displayed considerable cytotoxic activity against several cancer cell lines with IC50 values in the range of 1.2 to 5.0 microg/mL. The cytotoxic activity seemed to be related to DNA synthesis inhibition, as revealed by the reduction of 5-bromo-2'-deoxyuridine incorporation, and apoptosis induction, as indicated by the acridine orange/ethidium bromide assay and morphological changes after 24 h of incubation on leukemic cells.

  5. Inclusion complexation behavior of dyestuff guest molecules by a bridged bis(cyclomaltoheptaose)[bis(beta-cyclodextrin)] with a pyromellitic acid diamide tether.

    Science.gov (United States)

    Liu, Yu; Li, Li; Zhang, Heng-Yi; Liang, Peng; Wang, Hao

    2003-08-12

    A novel bridged bis(beta-cyclodextrin) with a pyromellitic acid 2,5-diamide tether (2) has been synthesized by reaction of 6(I)-(2-aminoethyleneamino)-6-deoxycyclomaltoheptaose [mono 6-(2-aminoethyleneamino)-6-deoxy-beta-cyclodextrin] with 1,2,4,5-benzenetetracarboxylic dianhydride. Its inclusion complexation behavior with some representative dyestuffs, i.e., Acridine Red (AR), Rhodamine B (RhB), Neutral Red (NR), Brilliant Green (BG), was studied by using UV-absorption, fluorescence, and 2D NMR spectroscopy. Fluorescence titrations have been performed at 25 degrees C in pH 7.2 buffer solution to calculate the binding constants of resulting complexes. These results obtained indicated that bis(beta-cyclodextrin) 2 exhibits the strongly enhanced binding ability with all dye molecules examined compared with natural cyclodextrins. The binding modes of 2 with dye molecules have been deduced by 2D NMR experiments to establish the correlations between molecular conformations and binding constants of inclusion complexation. It is found that the improved binding ability and molecular selectivity of 2 could be attributed to double-cavity cooperative inclusion interaction and the size/shape matching between the host and guest.

  6. DNA binding, cytotoxicity and inhibitory effect on RNA synthesis of two new 1-nitro-9-aminoacridine dimers.

    Science.gov (United States)

    Markovits, J; Wilmańska, D; Lescot, E; Studzian, K; Szmigiero, L; Gniazdowski, M

    1989-01-01

    Two 1-nitro-9-aminoacridine dimers were prepared: one bearing a spermine flexible linking chain, compound 4, the other a rigid dipiperidine-type linker, compound 7. Both dimers elicited a higher affinity constant for DNA than the parent monomeric drug nitracrine 2. This affinity was several orders lower than what was found for other dimeric compounds having the same linkers and no nitro group on the acridine ring (3, 5, 6 and 8). Bisintercalation was evidenced for compound 4 by viscosimetric measurements. In the absence of dithiothreitol, an inhibitory effect of RNA synthesis in vitro was observed for all the tested compounds except 2 and 7. In the presence of dithiothreitol, 4 and 7 formed irreversible complexes with DNA of decreased template properties. The level of the dimers binding was lower than that of the parent compound 2. Cross-links were detected by means of hydroxylapatite chromatography in a complex of the dimer bearing a flexible linking chain, compound 4 with DNA, while the compound 7-DNA complex eluted in the single-stranded DNA region. The extent of cytotoxicity of the two 1-nitro-9-aminoacridine dimers against L1210 cultured cells was different. PMID:2472225

  7. Transcription factors as targets for DNA-interacting drugs.

    Science.gov (United States)

    Gniazdowski, Marek; Denny, William A; Nelson, Stephanie M; Czyz, Malgorzata

    2003-06-01

    Gene expression, both tissue specific or inducible, is controlled at the level of transcription by various transcription factors interacting with specific sequences of DNA. Anticancer drugs and other potential therapeutic agents alter interactions of regulatory proteins with DNA by a variety of different mechanisms. The main ones, considered in the review, are: i) competition for the transcription factor DNA binding sequences by drugs that interact non-covalently with DNA (e.g. anthracyclines, acridines, actinomycin D, pyrrole antibiotics and their polyamide derivatives); ii) covalent modifications of DNA by alkylating agents (e.g. nitrogen mustards, cisplatin) that prevent transcription factors from recognizing their specific sequences, or that result in multiple "unnatural" binding sites in DNA which hijack the transcription factors, thus decreasing their availability in the nucleus; iii) competition with binding sites on the transcription factors by synthetic oligonucleotides or peptide nucleic acids in an antigene strategy. The latter compounds may also compete for binding sites on regulatory proteins, acting as decoys to lower their active concentration in the cell. In this review, we have summarized recent advances which have been made towards understanding the above mechanisms by which small molecules interfere with the function of transcription factors. PMID:12678680

  8. Effect of intercalating and groove-binding ligands on formation of covalent complexes between nitracrine (Ledakrin, C-283) or 8-methoxypsoralen and DNA.

    Science.gov (United States)

    Wilmańska, D; Małagocka, E; Szmigiero, L; Gniazdowski, M

    1984-07-18

    The effect of ethidium bromide, actinomycin D, distamycin A and netropsin on covalent binding of nitracrine (1-nitro-9-(3,3-N,N-dimethylaminopropylamino)acridine, Ledakrin, C-283) and 8-methoxypsoralen to DNA was examined. The competition was assayed either directly with [3H]- and [14C]nitracrine or indirectly by estimation of transcriptional template activity of nitracrine-DNA and 8-methoxypsoralen-DNA complexes formed in the presence of the ligands. A higher protective effect of ethidium bromide and distamycin on the photo-binding of 8-methoxypsoralen than on the dithiothreitol-dependent attachement of nitracrine to DNA assayed at 0.15 M KCl or NaCl was observed. The non-intercalating antibiotics showed lower competitive effect on 8-methoxypsoralen binding than ethidium bromide. Actinomycin D showed relatively low competition for both drugs with DNA. In contrast to the reaction of 8-methoxypsoralen, the decrease of nitracrine binding in the presence of competing ligands considerably depends on ionic strength. Particularly high inhibition of the adduct formation in the presence of ethidium at 1 M KCl was shown, while the amount of nitracrine bound in the presence of distamycin increases at elevated ionic strength. The results may indicate steric demands of the reaction between nitracrine and DNA. PMID:6733111

  9. Cytotoxic assay of endophytic fungus 1.2.11 secondary metabolites from Brucea javanica (L Merr towards cancer cell in vitro

    Directory of Open Access Journals (Sweden)

    Pratiwi Sudarmono

    2006-09-01

    Full Text Available Cytotoxic assay of secondary metabolite endophytic fungus 1.2.11 from Brucea javanica (L Merr has been carried out. Brucea javanica fruit collected from Cianjur was used in this experiment. Cytotoxic assay was done on Raji, NS-1, HeLa and Vero cells. The observation was done for 24 hours and also for 48 hours. IC50 was calculated using the Rich and Muench theory. To observe the working mechanism of cytotoxic process, DNA staining with etidium bromide and acridine orange was conducted. The cytotoxic assay of endophytic fungi 1.2.11 showed an IC50 of 58.35 μg/ml, 88.39 μg/ml on Raji cell,; 162.09 μg/ml, 66.24 μg/ml on NS cell; 361.21 μg/ml, 219.97 μg/ml on HeLa cell; and lastly 1075.18 μg/ml, 656.82 μg/ml on Vero cell after 24 and 48 hour incubation respectively. The results of this study showed that secondary metabolite of endophytic fungus 1.2.11 has selective cytotoxic effect towards cancer cell and also showed that it might cause apoptosis in NS-1cell. (Med J Indones 2006; 15:137-44 Keywords: Brucea javanica (L. Merr, endophytic microbe, Cytotoxic assay, endophytic isolate 1.2.11, apoptosis

  10. Identification keys on rattans (Calamus spp. from Central Sulawesi based on anatomical structure of stems

    Directory of Open Access Journals (Sweden)

    ANDI TANRA TELLU

    2005-04-01

    Full Text Available This study was conducted to obtain information the anatomical characteristics of 20 rattan species from Central Sulawesi and to use it for anatomical identification of rattan species. The rattan comprised 16 Calamus species, three Daemonorops species and one Korthalsia species. For anatomical observation 10-15 mm pieces of the mature stem from shares of tip do not have frond were processed with polyethilene glycol 2000, cut at 18-32 µm and stained with a combination of acridin-cryzoidin red and astrablue. Cleared preparation were used to observe stegmata, and macerated material was used to measure the length of fibers and vessel elements. Anilin sulfate was used to confirm the existence of lignin. Anatomical characteristics used in identification were shape and will thickening of epidermal cells and the position stomata at epidermal; the arrangement of sub epidermal parenchyma; composition of vascular bundles and their distribution; the shape and arrangement of central ground parenchyma and the occurrence of fiber bundles. The research result indicated that the anatomical character can be compiled to a key identify the rattan at genus and species level.

  11. Growth inhibition and apoptosis in cancer cells induced by polyphenolic compounds of Acacia hydaspica: Involvement of multiple signal transduction pathways

    Science.gov (United States)

    Afsar, Tayyaba; Trembley, Janeen H.; Salomon, Christine E.; Razak, Suhail; Khan, Muhammad Rashid; Ahmed, Khalil

    2016-01-01

    Acacia hydaspica R. Parker is known for its medicinal uses in multiple ailments. In this study, we performed bioassay-guided fractionation of cytotoxic compounds from A. hydaspica and investigated their effects on growth and signaling activity in prostate and breast cancer cell lines. Four active polyphenolic compounds were identified as 7-O-galloyl catechin (GC), catechin (C), methyl gallate (MG), and catechin-3-O-gallate (CG). The four compounds inhibited prostate cancer PC-3 cell growth in a dose-dependent manner, whereas CG and MG inhibited breast cancer MDA-MB-231 cell growth. All tested compounds inhibited cell survival and colony growth in both cell lines, and there was evidence of chromatin condensation, cell shrinkage and apoptotic bodies. Further, acridine orange, ethidium bromide, propidium iodide and DAPI staining demonstrated that cell death occurred partly via apoptosis in both PC-3 and MDA-MB-231 cells. In PC-3 cells treatment repressed the expression of anti-apoptotic molecules Bcl-2, Bcl-xL and survivin, coupled with down-regulation of signaling pathways AKT, NFκB, ERK1/2 and JAK/STAT. In MDA-MB-231 cells, treatment induced reduction of CK2α, Bcl-xL, survivin and xIAP protein expression along with suppression of NFκB, JAK/STAT and PI3K pathways. Our findings suggest that certain polyphenolic compounds derived from A. hydaspica may be promising chemopreventive/therapeutic candidates against cancer. PMID:26975752

  12. Cytocompatibility, cytotoxicity and genotoxicity analysis of dental implants

    Science.gov (United States)

    Reigosa, M.; Labarta, V.; Molinari, G.; Bernales, D.

    2007-11-01

    Several types of materials are frequently used for dental prostheses in dental medicine. Different treatments with titanium are the most used. The aim of the present study was to analyze by means of cytotoxicity and cytocompatibility techniques the capacity of dental implants to integrate to the bone tissue. Cultures of UMR 106 cell line derived from an osteosarcoma were used for bioassays mainly because they show many of the properties of osteoblasts. Dental implant samples provided by B&W company were compared with others of recognized trademarks. The first ones contain ASTM titanium (8348 GR2) with acid printing. Cytotoxicity was analyzed by means of lysosome activity, using the neutral red technique and alkaline phosphatase enzyme activity. Cell variability was determined by means of the acridine ethidium-orange bromide technique. One-way ANOVA and Bonferroni and Duncan post-ANOVA tests were used for the statistical analysis. The assays did not show significant differences among the dental implants analyzed. Our findings show that the dental prostheses studied present high biocompatibility, quantified by the bioassays performed. The techniques employed revealed that they can be a useful tool for the analysis of other materials for dental medicine use.

  13. Cytocompatibility, cytotoxicity and genotoxicity analysis of dental implants

    Energy Technology Data Exchange (ETDEWEB)

    M, Reigosa [Instituto Multidisciplinario de BiologIa Celular (IMBICE) CONICET-CIC.C.C. 403 (1900) La Plata. (Argentina); V, Labarta [Instituto Multidisciplinario de BiologIa Celular (IMBICE) CONICET-CIC.C.C. 403 (1900) La Plata. (Argentina); G, Molinari [Catedra de CitologIa.Facultad de Ciencias Naturales y Museo.UNLP (Argentina); D, Bernales

    2007-11-15

    Several types of materials are frequently used for dental prostheses in dental medicine. Different treatments with titanium are the most used. The aim of the present study was to analyze by means of cytotoxicity and cytocompatibility techniques the capacity of dental implants to integrate to the bone tissue. Cultures of UMR 106 cell line derived from an osteosarcoma were used for bioassays mainly because they show many of the properties of osteoblasts. Dental implant samples provided by B and W company were compared with others of recognized trademarks. The first ones contain ASTM titanium (8348 GR2) with acid printing. Cytotoxicity was analyzed by means of lysosome activity, using the neutral red technique and alkaline phosphatase enzyme activity. Cell variability was determined by means of the acridine ethidium-orange bromide technique. One-way ANOVA and Bonferroni and Duncan post-ANOVA tests were used for the statistical analysis. The assays did not show significant differences among the dental implants analyzed. Our findings show that the dental prostheses studied present high biocompatibility, quantified by the bioassays performed. The techniques employed revealed that they can be a useful tool for the analysis of other materials for dental medicine use.

  14. Nephrotoxicity of Bence-Jones proteins: interference in renal epithelial cell acidification

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    Nicastri A.L.

    2002-01-01

    Full Text Available The aim of the present study was to evaluate the acidification of the endosome-lysosome system of renal epithelial cells after endocytosis of two human immunoglobulin lambda light chains (Bence-Jones proteins, BJP obtained from patients with multiple myeloma. Renal epithelial cell handling of two BJP (neutral and acidic BJP was evaluated by rhodamine fluorescence. Renal cells (MDCK were maintained in culture and, when confluent, were incubated with rhodamine-labeled BJP for different periods of time. Photos were obtained with a fluorescence microscope (Axiolab-Zeiss. Labeling density was determined on slides with a densitometer (Shimadzu Dual-Wavelength Flying-Spot Scanner CS9000. Endocytosis of neutral and acidic BJP was correlated with acidic intracellular compartment distribution using acridine orange labeling. We compared the pattern of distribution after incubation of native neutral and acidic BJP and after complete deglycosylation of BJP by periodate oxidation. The subsequent alteration of pI converted neutral BJP to acidic BJP. There was a significant accumulation of neutral BJP in endocytic structures, reduced lysosomal acidification, and a diffuse pattern of acidification. This pattern was reversed after total deglycosylation and subsequent alteration of the pI to an acidic BJP. We conclude that the physicochemical characteristics of BJP interfere with intracellular acidification, possibly explaining the strong nephrotoxicity of neutral BJP. Lysosomal acidification is fundamental for adequate protein processing and catabolism.

  15. Effect of all-trans retinoic acid 0n drug sensitivity and expression of survivin in LoVo cells

    Institute of Scientific and Technical Information of China (English)

    2008-01-01

    Background All-trans retinoic acid(ATRA)can influence the tumor cell proliferation cycle,and some chemotherapeutic drugs are cycle specific.In this study,we hypothesize that ATRA can enhance chemotherapeutic drug sensitivity by affecting the cell cycle of tumor cells.Methods The cell cycle of LoVo cells was evaluated using flow cytometry(FCM).Cell viability was analyzed using the MTT assay.The morphologic changes in the treated LoVo cells were measured with acridine orange (AO)/ethidium bromide(EB)staining.Expression of survivin in LoVo cells was analyzed by immunofluorescence assay.Results After LoVo cells were treated with ATRA,the G0/G1 ratio of the tumor cells increased and the cell ratio of Sand G2/M-phase decreased.Viability of the cells decreased significantly after combined treatment with ATRA and 5-fluorouracil(5-FU)or mitomycin c(MMC) and was evaluated by fluorescence microscopy.Expression level of survivin in the tumor cells decreased after ATRA combination treatment.Conclusions ATRA enhances drug sensitivity of the LoVo cell line to cell cycle-specific agents and inhibits the expression of survivin in LoVo cells.The combination of ATRA and 5-FU or MMC promoted cell apoptosis,and the mechanism involved in apoptosis may be related to inhibition of survivin gene expression.

  16. Toxicity of silver nanoparticles in zebrafish models

    Energy Technology Data Exchange (ETDEWEB)

    Asharani, P V; Valiyaveettil, Suresh [Department of Chemistry, Faculty of Science, National University of Singapore, 3 Science Drive 3, 117543 (Singapore); Wu Yilian; Gong Zhiyuan [Department of Biological Sciences, National University of Singapore, Science Drive 4, 117543 (Singapore)], E-mail: chmsv@nus.edu.sg

    2008-06-25

    This study was initiated to enhance our insight on the health and environmental impact of silver nanoparticles (Ag-np). Using starch and bovine serum albumin (BSA) as capping agents, silver nanoparticles were synthesized to study their deleterious effects and distribution pattern in zebrafish embryos (Danio rerio). Toxicological endpoints like mortality, hatching, pericardial edema and heart rate were recorded. A concentration-dependent increase in mortality and hatching delay was observed in Ag-np treated embryos. Additionally, nanoparticle treatments resulted in concentration-dependent toxicity, typified by phenotypes that had abnormal body axes, twisted notochord, slow blood flow, pericardial edema and cardiac arrhythmia. Ag{sup +} ions and stabilizing agents showed no significant defects in developing embryos. Transmission electron microscopy (TEM) of the embryos demonstrated that nanoparticles were distributed in the brain, heart, yolk and blood of embryos as evident from the electron-dispersive x-ray analysis (EDS). Furthermore, the acridine orange staining showed an increased apoptosis in Ag-np treated embryos. These results suggest that silver nanoparticles induce a dose-dependent toxicity in embryos, which hinders normal development.

  17. 7-Hydroxydehydronuciferine induces human melanoma death via triggering autophagy and apoptosis.

    Science.gov (United States)

    Wu, Pei-Fang; Chiu, Chien-Chih; Chen, Chung-Yi; Wang, Hui-Min David

    2015-12-01

    Melanoma is the deadliest cancer. We identified 7-hydroxydehydronuciferine (7-HDNF) isolated from the leaves of Nelumbo nucifera Gaertn cv. Rosa-plena to be a bio-active agent that antagonizes melanoma tumor growth in mice xenograft model in vivo. Cell proliferation assay demonstrated strong anticancer effects of 7-HDNF to exhibit a dose-dependent behaviour and displayed minor cytotoxicities on normal human skin cells, including epidermal keratinocytes and melanocytes, and dermal fibroblasts. With acridine orange (AO) staining and flow analysis, we found 7-HDNF induced the formation of intracellular vacuoles and the augmentation of acidic vesicular organelles (AVO). The apoptotic cell death ratio was measured via two-dimensional flow cytometry by annexin V-fluorescein isothiocyanate (FITC)/propidium iodide (PI) double stained to confirm the cellular membrane asymmetry lost. One-dimensional flow cytometric analysis showed 7-HDNF increased the cellular arrest in cell cycle at the G2/M phase. Through Western blot examinations, protein expressions were discovered to verify autophagy and apoptosis response mechanisms sharing the associated pathways. Finally, 7-HDNF presented a high-quality antimigratory activity in wound-healing assay. Overall, 7-HDNF presented high-quality anticancer bio-functions and inhibited melanoma tumor growth in vivo and in vitro. PMID:26174122

  18. Atorvastatin Protects Vascular Smooth Muscle Cells From TGF-β1-Stimulated Calcification by Inducing Autophagy via Suppression of the β-Catenin Pathway

    Directory of Open Access Journals (Sweden)

    Demin Liu

    2014-01-01

    Full Text Available Background: Arterial calcification is a major event in the progression of atherosclerosis. It is reported that statins exhibit various protective effects against vascular smooth muscle cell (VSMC inflammation and proliferation in cardiovascular remodeling. Although statins counteract atherosclerosis, the molecular mechanisms of statins on the calcium release from VSMCs have not been clearly elucidated. Methods: Calcium content of VSMCs was measured using enzyme-linked immunosorbent assay (ELISA. The expression of proteins involved in cellular transdifferentiation was analyzed by western blot. Cell autophagy was measured by fluorescence microscopic analysis for acridine orange staining and transmission electron microscopy analysis. The autophagic inhibitors (3-MA, chloroquine, NH4Cl and bafilomycin A1 and β-catenin inhibitor JW74 were used to assess the effects of atorvastatin on autophagy and the involvement of β-catenin on cell calcification respectively. Furthermore, cell transfection was performed to overexpress β-catenin. Results: In VSMCs, atorvastatin significantly suppressed transforming growth factor-β1 (TGF-β1-stimulated calcification, accompanied by the induction of autophagy. Downregulation of autophagy with autophagic inhibitors significantly suppressed the inhibitory effect of atorvastatin on cell calcification. Moreover, the beneficial effect of atorvastatin on calcification and autophagy was reversed by β-catenin overexpression. Conversely, JW74 supplement enhanced this effect. Conclusion: These data demonstrated that atorvastatin protect VSMC from TGF-β1-stimulated calcification by inducing autophagy through suppression of the β-catenin pathway, identifying autophagy induction might be a therapeutic strategy for use in vascular calcification.

  19. Etiology and Evaluation of Sperm Chromatin Anomalies

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    Marziyeh Tavalaee

    2008-01-01

    Full Text Available Evidence suggests that human sperm chromatin anomalies adversely affect reproductive outcomesand infertile men possess substantially amount of sperm with chromatin anomalies than fertilemen.Routine semen analysis evaluates parameters such as sperm motility and morphology, but doesnot examine the nuclear DNA integrity of spermatozoa. It has been suggested that altered nuclearchromatin structure or damaged DNA in spermatozoa could modify the special cellular functionsof human spermatozoa, and thereby affect the fertility potential. Intra-cytoplasmic sperm injection(ICSI bypass the barriers to fertilization for such a sperm, then the effect of chromatin anomalies onthe development remains a concern. Therefore, it is essential to develop and use accurate diagnostictests, which may provide better prognostic capabilities than the standard sperm assessments. Thisreview discusses our current understanding of the structure and organization of sperm DNA,the different procedures for assessment of sperm chromatin anomalies including comet assay,Chromomycin A3 (CMA3, sperm chromatin structure assay (SCSA, acridine orange test (AOT,terminal TdT-mediated dUTP-nick-end labelling (TUNEL assay, aniline blue and sperm chromatindispersion (SCD test and the impact of chromatin anomalies on reproductive outcome.

  20. Fibrates inhibit the apoptosis of Batten disease lymphoblast cells via autophagy recovery and regulation of mitochondrial membrane potential.

    Science.gov (United States)

    Hong, Minho; Song, Ki Duk; Lee, Hak-Kyo; Yi, SunShin; Lee, Yong Seok; Heo, Tae-Hwe; Jun, Hyun Sik; Kim, Sung-Jo

    2016-03-01

    Batten disease (BD; also known as juvenile neuronal ceroid lipofuscinosis) is a genetic disorder inherited as an autosomal recessive trait and is characterized by blindness, seizures, cognitive decline, and early death resulting from the inherited mutation of the CLN3 gene. Mitochondrial oxidative stress, endoplasmic reticulum (ER) stress, disrupted autophagy, and enhanced apoptosis have been suggested to play a role in BD pathogenesis. Fibrates, a class of lipid-lowering drugs that induce peroxisome proliferator-activated receptor-α (PPAR-α) activation, are the most commonly used PPAR agonists. Assuming that fibrates have a neuroprotective effect, we studied the effects of fibrates, fenofibrate, bezafibrate, and gemfibrozil on apoptosis, depolarization of mitochondrial membrane, and defective autophagy in BD lymphoblast cells. The viability of fibrate-treated BD lymphoblast cells increased to levels of normal lymphoblast cells. In addition, treatment with fibrates inhibited depolarization of mitochondrial membrane potential in BD lymphoblast cells. Defective autophagy in BD lymphoblast cells was normalized when treated with fibrates as indicated by increased acridine orange staining. The recovery of autophagy in BD lymphoblast cells is most likely attributed to the upregulation of autophagy proteins, lysosomal-associated membrane protein 1 (LAMP1), and LC3 I/II, after treatment with fibrates. This study therefore suggests that fibrates may have a therapeutic potential against BD. PMID:26659390

  1. Cysteine: A Novel Neural Inducer for Rat Bone Marrow Mesenchymal Stem Cells

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    Malek Soleimani Mehranjani

    2014-06-01

    Full Text Available Objective: Mesenchymal stem cells (MSCs can differentiate into various cell types. Since cysteine has structural similarities to neuronal inducers β-mercaptoethanol and glutathione, we examined its effect on neural induction of rat bone marrow MSCs. Materials and Methods: In this experimental study, cells were treated in a medium containing 1mM cysteine for 24 hours prior to treatment with neuron inducing medium containing 10 mM cysteine for 1, 2 and 3 hours. Cell viability and morphology were assessed by 3-(4,5-dimethylthiazol-2-Yl-2,5-diphenyltetrazolium bromide (MTT assay and, Hoechst, propidium iodide and acridine orange staining respectively. Expression of nestin and β-Tubulin III genes, as neural cell-specific markers, was studied reverse transcription polymerase chain reaction (RT-PCR. The data was statistically analyzed using One-Way ANOVA and Tukey’s test and p<0.05 was considered significant. Results: After 3 hours of treatment, neuron like morphology with a considerable expression of nestin and β-Tubulin III genes was apparent. The mean cell viability was not significantly different at 1, 2 and 3 hours following induction, compared with the control cells. Conclusion: Cysteine can induce neural features in rat bone marrow MSCs without reducing cell viability. Therefore, it can be considered as a safer alternative to toxic neural inducer agents such as β-mercaptoethanol.

  2. Environmental & lifestyle factors in deterioration of male reproductive health

    Directory of Open Access Journals (Sweden)

    Sunil Kumar

    2014-01-01

    Full Text Available Background & objectives: Male reproductive function in the general population has been receiving attention in recent years due to reports of various reproductive and developmental defects, which might be associated with various lifestyle and environmental factors. This study was carried out to determine the role of various lifestyle and environmental factors in male reproduction and their possible association with declining semen quality, increased oxidative stress as well as sperm DNA damage. Methods: Semen samples were obtained from 240 male partners of the couples consulting for infertility problem. Semen analysis was carried out using WHO criteria and subjects were categorized on the basis of self reported history of lifestyle as well as environmental exposure. The oxidative and antioxidant markers; lipid peroxidation (LPO, superoxide dismutase (SOD and catalase (CAT as well as DNA damage by acridine orange test (AO were determined. Results: The presence of abnormal semen parameters was significantly higher among the lifestyle and/or environmental exposed subjects as compared to the non-exposed population. Further, the levels of antioxidants were reduced and sperm DNA damage was more among the lifestyle and/or environmental exposed subjects, though the changes were not significant. Interpretation & conclusions: These findings indicated that various lifestyle factors such as tobacco smoking, chewing and alcohol use as well as exposure to toxic agents might be attributed to the risk of declining semen quality and increase in oxidative stress and sperm DNA damage.

  3. Detection on emamectin benzoate-induced apoptosis and DNA damage in Spodoptera frugiperda Sf-9 cell line.

    Science.gov (United States)

    Wu, Xiwei; Zhang, Lei; Yang, Chao; Zong, Mimi; Huang, Qingchun; Tao, Liming

    2016-01-01

    Emamectin benzoate (EMB), an important macrocyclic lactone insecticide that belongs to the avermectin family and possesses excellent potency in controlling pests, is non-carcinogenic and non-mutagenic conducted in rats and mice, but EMB-induced cytotoxicity and genotoxicity in arthropod insect have been seldom reported yet. In the present paper, we quantified the cytotoxicity of EMB through the detections on cell viability, DNA damage, and cell apoptosis in Spodoptera frugiperda Sf-9 cells in vitro. The results showed that EMB caused a concentration- and time-dependent reduction on the viability of Sf-9 cells, and the median inhibitory concentrations (IC50) were 3.34μM at 72h of exposure. The dual acridine orange/ethidium bromide staining showed that exposure to EMB induced a significant time- and concentration-dependent increase on cell apoptosis. The alkaline comet assay revealed that EMB induced significant increases on single-strand DNA breaks, and the percentage of γH2AX-positive cells represented a time- and concentration-dependent formation of DNA double-strand breaks in Sf-9 cells. Interestingly, the similar cytotoxic actions of EMB also went for the human cancerous HeLa cells as a control cell group. Data demonstrated the potential cytotoxic effect of EMB on Sf-9 cells that was significantly greater than the effect of hydrogen peroxide at the same concentrations.

  4. Astragalus extract inhibits destruction of gastric cancer cells to mesothelial cells by anti-apoptosis

    Institute of Scientific and Technical Information of China (English)

    Di Na; Fu-Nan Liu; Zhi-Feng Miao; Zong-Min Du; Hui-Mian Xu

    2009-01-01

    AIM: To determine the inhibitory effect of Astragalus memebranaceushas on gastric cancer cell supernatantinduced apoptosis of human peritoneal mesothelial cells. METHODS: Human peritoneal mesothelial cell (HPMC) line HMrSV5 was co-incubated with gastric cancer cell supernatant (MKN45) and/or Astragalus memebranaceushas. Morphological changes in gastric cancer cells were observed under phase-contrast microscope. Quantitative cell damage was determined by MTT assay. Apoptosis was determined under transmission electron microscope and quantified by detecting acridine orange/ethidium bromide-stained (AO/EB) condensed nuclei under fluorescent microscope or by flow cytometry. Expressions of Bcl-2 and Bax were evaluated with immunostaining. RESULTS: Morphological changes and exfoliation occurred and naked areas appeared in cultured HMrSV5 cells 24 h after they were treated with gastric cancer cell supernatant. Cell supernatant from MKN45 gastric cancer cells induced apoptosis of HMrSV5 cells in a time-dependent manner. Obvious morphological changes were observed in cell apoptosis, such as condensation of chromatin, nuclear fragmentations and apoptotic bodies. Astragalus memebranaceus could partly suppress these changes and regulate the expressions of Bcl-2 and Bax in HMrSV5 cells. CONCLUSION: Gastric cancer cells induce apoptosis of HPMCs through the supernatant. Astragalus memebranaceushas inhibits this phenomenon and can be used an adjuvant chemothera-peutic agent in gastric cancer therapy.

  5. Preferentially Cytotoxic Constituents of Andrographis paniculata and their Preferential Cytotoxicity against Human Pancreatic Cancer Cell Lines.

    Science.gov (United States)

    Lee, Sullim; Morita, Hiroyuki; Tezuka, Yasuhiro

    2015-07-01

    In the course of our search for anticancer agents based on a novel anti-austerity strategy, we found that the 70% EtOH extract of the crude drug Andrographis Herba (aerial parts of Andrographis paniculata), used in Japanese Kampo medicines, killed PANC-1 human pancreatic cancer cells preferentially in nutrient-deprived medium (NDM). Phytochemical investigation of the 70% EtOH extract led to the isolation of 21 known compounds consisting of six labdane-type diterpenes (11, 15, 17-19, 21), six flavones (5, 7, 10, 12, 14, 20), three flavanones (2, 6, 16), two sterols (3, 8), a fatty acid (1), a phthalate (4), a triterpene (9), and a monoterpene (13). Among them, 14-deoxy-11,12-didehydroandrographolide (17) displayed the most potent preferential cytotoxicity against PANC-1 and PSN-1 cells with PC50 values of 10.0 μM and 9.27 μM, respectively. Microscopical observation, double staining with ethidium bromide (EB) and acridine orange (AO), and flow cytometry with propidium iodide/annexin V double staining indicated that 14-deoxy-11,12-didehydroandrographolide (17) triggered apoptosis-like cell death in NDM with an amino acids and/or serum-sensitive mode. PMID:26410998

  6. Positive selection of mutants with cell envelope defects of a Salmonella typhimurium strain hypersensitive to the products of genes hisF and hisH. [uv radiation

    Energy Technology Data Exchange (ETDEWEB)

    Anton, D.N.

    1979-03-01

    Strain SB564 and its derivative DA78 are hypersensitive to the inhibitory action of the proteins coded for by genes hisF and hisH on cell division. Transduction of hisO1242, a regulatory mutation that elicits a very high level of expression of the histidine operon, into these strains resulted in the production of long filamentous cells carrying large balloons and in growth failure. Forty-one hisO1242 derivatives that escaped inhibition were isolated. These strains showed a large variety of alterations, many of which were related to the cell envelope. The more-frequent alterations included: changes in cell shape, increased sensitivity to one or more of several drugs (deoxycholate, cycloserine, penicillin, novobiocin, acridine orange), increased autolytic activity in alkaline buffer, anomalous fermentation of maltose on eosin--methylene blue plates, and temperature-conditional cell division. The alterations are produced, in some of the strains, by pleiotropic mutations in gene envB. Strains affected in divC, divD, and rodA loci have also been identified. Genetic analaysis has shown that several strains carry more than one envelope mutation. It is assumed that envelope mutations are positively selected because they somehow alleviate the particularly severe inhibition of cell division caused, in strains SB564 and DA78, by the excessive synthesis of hisF and hisH gene products.

  7. STUDY OF BACTERIAL RESISTANCE TO ORGANOPHOSPHOROUS PESTICIDES IN IRAN

    Directory of Open Access Journals (Sweden)

    A. Nazarian and M. Mousawi

    2005-07-01

    Full Text Available The broadness application of organophosphorus compounds has abounded the number of its polluted areas. Bioremediation has widely focused on insitu bacterial degradation of organophosphorus residues in the world. Therefore, in this research six numbers of samples from two different sources, soil and water randomly were isolated using different organophosphorus pesticides containing mineral solution without supplementation. More than 100 isolated strains were selected according to their simultaneous optimal growth on mineral medium with organophosphorus and Mac Conkey,s agar. More than 50 percent of them were lost above resistance. The resistant strains were identified by two methods, the biochemical convention and API 20E procedure with positive agreement. The identified strains belonged to Pseudomonas and Flavobacterium species. The maximum tolerant concentrations of different organophosphorus pesticides by these resistant strains were 2.5, 4 and 8 g/L of guthion, methyl parathion and Dimethoate, respectively. The resistance to these pesticides due to organ phosphorous degrading plasmids had the ability to express hydrolytic enzymes. Resistant bacteria lost these plasmids by acridin orange and could translocate to sensitive strains. Thus, certain environmental bacteria could be used as protection tools against antinerve agents.

  8. Antioxidant, Antimicrobial and Antiproliferative Activities of Five Lichen Species

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    Snežana Marković

    2011-08-01

    Full Text Available The antioxidative, antimicrobial and antiproliferative potentials of the methanol extracts of the lichen species Parmelia sulcata, Flavoparmelia caperata, Evernia prunastri, Hypogymnia physodes and Cladonia foliacea were evaluated. The total phenolic content of the tested extracts varied from 78.12 to 141.59 mg of gallic acid equivalent (GA/g of extract and the total flavonoid content from 20.14 to 44.43 mg of rutin equivalent (Ru/g of extract. The antioxidant capacities of the lichen extracts were determined by 2,2-diphenyl-1-picrylhydrazyl (DPPH radicals scavenging. Hypogymnia physodes with the highest phenolic content showed the strongest DPPH radical scavenging effect. Further, the antimicrobial potential of the lichen extracts was determined by a microdilution method on 29 microorganisms, including 15 strains of bacteria, 10 species of filamentous fungi and 4 yeast species. A high antimicrobial activity of all the tested extracts was observed with more potent inhibitory effects on the growth of Gram (+ bacteria. The highest antimicrobial activity among lichens was demonstrated by Hypogymnia physodes and Cladonia foliacea. Finally, the antiproliferative activity of the lichen extracts was explored on the colon cancer adenocarcinoma cell line HCT-116 by MTT (3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyltetrazolium bromide viability assay and acridine orange/ethidium bromide staining. The methanol extracts of Hypogymnia physodes and Cladonia foliacea showed a better cytotoxic activity than the other extracts. All lichen species showed the ability to induce apoptosis of HCT-116 cells.

  9. Development of a simple, sensitive and inexpensive ion-pairing cloud point extraction approach for the determination of trace inorganic arsenic species in spring water, beverage and rice samples by UV-Vis spectrophotometry.

    Science.gov (United States)

    Gürkan, Ramazan; Kır, Ufuk; Altunay, Nail

    2015-08-01

    The determination of inorganic arsenic species in water, beverages and foods become crucial in recent years, because arsenic species are considered carcinogenic and found at high concentrations in the samples. This communication describes a new cloud-point extraction (CPE) method for the determination of low quantity of arsenic species in the samples, purchased from the local market by UV-Visible Spectrophotometer (UV-Vis). The method is based on selective ternary complex of As(V) with acridine orange (AOH(+)) being a versatile fluorescence cationic dye in presence of tartaric acid and polyethylene glycol tert-octylphenyl ether (Triton X-114) at pH 5.0. Under the optimized conditions, a preconcentration factor of 65 and detection limit (3S blank/m) of 1.14 μg L(-1) was obtained from the calibration curve constructed in the range of 4-450 μg L(-1) with a correlation coefficient of 0.9932 for As(V). The method is validated by the analysis of certified reference materials (CRMs).

  10. Pro-apoptotic effect of Persea americana var. Hass (avocado) on Jurkat lymphoblastic leukemia cells.

    Science.gov (United States)

    Bonilla-Porras, Angelica R; Salazar-Ospina, Andrea; Jimenez-Del-Rio, Marlene; Pereañez-Jimenez, Andres; Velez-Pardo, Carlos

    2013-11-01

    Abstract Context: Therapy for leukemia has a limited efficacy. There is a need to search for alternative anti-leukemia therapies. Persea americana Mill var. Hass (Lauraceae) is a tropical fruit (avocado) that might be used against cancer. Objective: To investigate whether P. americana induces death in Jurkat lymphoblastic leukemia cells. Materials and methods: Four ethanol extracts (0.1, 0.5, 1, 2 and 5 mg/mL) from avocado fruit (endocarp, whole seed, seed and leaves) were analyzed against Jurkat cells. Hydrogen peroxide generation by oxidation of 2',7'-dichlorodihydrofluorescein diacetate to the fluorescent compound 2',7'-dichlorfluorescein assay, acridine orange/ethidium bromide staining, flow cytometry analysis of annexin-V/7-amino-actinomycin, mitochondrial membrane potential and immunocytochemistry detection of transcription factor p53, caspase-3 and apoptosis-inducing factor (AIF) were evaluated. Results: Endocarp, seed, whole seed, and leaf (0.1 mg/mL) extracts induced significant apoptosis in Jurkat cells (p avocado and its therapeutic action on leukemia.

  11. Protective effects of Rheum tanguticum polysaccharide against hydrogen peroxide-induced intestinal epithelial cell injury

    Institute of Scientific and Technical Information of China (English)

    Lin-Na Liu; Qi-Bing Mei; Li Liu; Feng Zhang; Zhen-Guo Liu; Zhi-Peng Wang; Ru-Tao Wang

    2005-01-01

    AIM: To describe the effect of Rheum tanguticum polysaccharide (RTP) on hydrogen peroxide-induced human intestinal epithelial cell injury.METHODS: Hydrogen peroxide (100 μmol/L) was introduced to induce human intestinal epithelial cell injury.Cells were pretreated with RTP (30,100,300 μg/mL) for 24 h before exposure to hydrogen peroxide. Cell viability was detected by MTr assay and morphological observation.Acridine orange staining and flow cytometry were performed to assess cell apoptosis. Lactate dehydrogenase (LDH) activity, production of malondialdehyde (MDA) and superoxide dismutase (SOD) activity were measured by spectrophotometry with corresponding assay kits.RESULTS: Following exposure to H2O2, a marked decrease in cell survival and SOD activity, increased production of MDA, LDH leakage and cell apoptosis were found.Pretreatment of the cells with RTP could significantly elevate cell survival, SOD activity and decrease the level of MDA, LDH activity and cell apoptosis.CONCLUSION: RTP may have cytoprotective and antioxidant effects against H2O2-induced intestinal epithelial cell injury by inhibiting cell apoptosis and necrosis. This might be one of the possible mechanisms of RTP for the treatment of ulcerative colitis in rats.

  12. Apoptosis induced by diallyl disulfide in human breast cancer cell line MCF.71

    Institute of Scientific and Technical Information of China (English)

    Xiao-yong LEI; Shu-qiong YAO; Xu-yu ZU; Ze-xiang HUANG; Li-juan LIU; Miao ZHONG; Bing-yang ZHU; Sheng- song TANG; Duan-fang LIAO

    2008-01-01

    Aim:To investigate the effect of diallyl disulfide (DADS),a component of garlic,on apoptosis in human mammary cancer cell line (MCF-7) and its mechanisms.Methods:Cytotoxicity was analyzed by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assays.Morphology of apoptotic cells was detected by acridine orange and ethidium bromide staining.Apoptotic cells stained with propidium iodide were examined using flow cytometry.Protein levels were detected by Western blot analysis.Results:DADS inhibited the proliferation of MCF-7 cells and induced the apoptotic ratio to increase rapidly.Cleavage of the caspase-3 and caspase-3 substrate poly(ADP-ribose) polymerase was observed in MCF-7 cells after 24 h of treatment with DADS.When the MCF-7 cells were signal-regulated kinase (ERK),a mitogen-activated protein kinase,was inhibited after 6 h; court N-terminal kinase (JNK),that is stress-activated protein kinase (SAPK),and p38 mitogen-aetivated protein kinase were activated after 6 h.Conclusion:These results suggest that DADS both inhibits the proliferation of MCF-7 cells and induces apoptosis of MCF-7 cells.The mechanisms may include the inhibition of ERK and the activation of the SAPK/JNK and p38 pathways.

  13. Role of hesperetin (a natural flavonoid) and its analogue on apoptosis in HT-29 human colon adenocarcinoma cell line--a comparative study.

    Science.gov (United States)

    Sivagami, Gunasekaran; Vinothkumar, Rajamanickam; Bernini, Roberta; Preethy, Christo Paul; Riyasdeen, Anvarbatcha; Akbarsha, Mohammad Abdulkader; Menon, Venugopal Padmanaban; Nalini, Namasivayam

    2012-03-01

    Colon cancer is one of the serious health problems in most developed countries and its incidence rate is increasing in India. Hesperetin (HN) (3',5,7-trihydroxy-4'-methoxyflavonone) and hesperetin analogue (HA) were tested for their apoptosis inducing ability. Methyl thiazolyl tetrazolium assay revealed a dose as well as duration-dependent reduction of HT-29 (colon adenocarcinoma) cellular growth in response to HN and HA treatment. At 24 h 70 μM of HN and 32 μM of HA showed 50% reduction of HT-29 cellular growth. Acridine orange/ethidium bromide staining showed apoptotic features of cell death induced by HN and HA. Rhodamine 123 staining showed significant reduction in mitochondrial membrane potential induced by HN and HA. HN and HA induced DNA damage was confirmed by comet tail formation. Lipid peroxidation markers (TBARS) and protein oxidation marker (PCC) were significantly elevated in HN and HA treated groups. Enzymic antioxidants such as superoxide dismutase (SOD), catalase (CAT), glutathione peroxidase (GPx) were slightly decreased in their activities compared to control (untreated HT-29 cells). Results of Western blot analysis of apoptosis associated genes revealed an increase in cytochrome C, Bax, cleaved caspase-3 expression and a decrease in Bcl-2 expression. These findings indicate that HN and HA induce apoptosis on HT-29 via Bax dependent mitochondrial pathway involving oxidant/antioxidant imbalance. PMID:22142698

  14. Gram-typing of mastitis bacteria in milk samples using flow cytometry

    DEFF Research Database (Denmark)

    Langerhuus, Sine Nygaard; Ingvartsen, Klaus Lønne; Bennedsgaard, Torben Werner;

    2013-01-01

    Fast identification of pathogenic bacteria in milk samples from cows with clinical mastitis is central to proper treatment. In Denmark, time to bacterial diagnosis is typically 24 to 48 h when using traditional culturing methods. The PCR technique provides a faster and highly sensitive identifica......Fast identification of pathogenic bacteria in milk samples from cows with clinical mastitis is central to proper treatment. In Denmark, time to bacterial diagnosis is typically 24 to 48 h when using traditional culturing methods. The PCR technique provides a faster and highly sensitive...... cytometry-based method, which can detect and distinguish gram-negative and gram-positive bacteria in mastitis milk samples. The differentiation was based on bacterial fluorescence intensities upon labeling with biotin-conjugated wheat germ agglutinin and acridine orange. Initially 19 in-house bacterial...... characteristic curves for the 19 bacterial cultures. The method was then tested on 53 selected mastitis cases obtained from the department biobank (milk samples from 6 gram-negative and 47 gram-positive mastitis cases). Gram-negative bacteria in milk samples were detected with a sensitivity of 1...

  15. A novel method for the determination of fast green in grape wine based on resonance Rayleigh scattering

    Science.gov (United States)

    Li, Qin; Tan, Xuanping; Zheng, Xiaobo; Tang, Weiwei; Yang, Jidong

    2015-11-01

    A novel resonance Rayleigh scattering method was developed for the determination of fast green (FCF) in grape wine. In pH 2.5 Britton Robinson (BR) buffer solution, the scattering signal of acridine orange (AO) was remarkably enhanced after adding trace amount of FCF and forming an ion-association complex, which not only resulted in the change of absorption spectrum, fluorescence spectra, but also led to a significant enhancement of resonance Rayleigh scattering (RRS), frequency doubling scattering (FDS), and second order scattering (SOS). The linear ranges and detection limits for RRS, SOS and FDS were 2-45 × 10-6 mol L-1, 2-24 × 10-6 mol L-1, 2-20 × 10-6 mol L-1, and 8.0 × 10-8 mol L-1, 4.7 × 10-7 mol L-3, 1.0 × 10-7 mol L-3, respectively. In this work, the optimum conditions, the influencing factors and the effects of coexisting substances on the reaction were investigated. The method can be applied to the determination of FCF in grape wine and the results were satisfactory.

  16. Differences in dispersion of influenza virus lipids and proteins during fusion.

    Science.gov (United States)

    Lowy, R J; Sarkar, D P; Whitnall, M H; Blumenthal, R

    1995-02-01

    Digitally enhanced low-light-level fluorescence video microscopy and immunochemical staining were used to examine influenza virus envelope lipid and protein redistribution during pH-induced fusion. Video microscopy was performed using viruses labeled with either the lipid analogue octadecylrhodamine B (R18) or fluorescein isothiocyanate (FITC) covalently linked to envelope proteins. Viruses were bound to human red blood cells, and the pattern and intensity of fluorescence were monitored for 30 min while cell-virus complexes were perfused with pH 7.4 or 4.8 media at temperatures either above or below 20 degrees C. R18 showed complete redistribution and dequenching by 30 min at all incubation temperatures, confirming reports that viral fusion occurs at subphysiological temperatures. FITC-labeled protein showed spatial redistribution at 28 degrees C but no change at low temperature. Electron microscopy observations of immunochemical staining of viral proteins confirmed both that protein redistribution at 37 degrees C was slower than R18 and the failure of movement within 30 min at 16 degrees C. Video microscopy monitoring of RNA staining by acridine orange of virus-cell complexes showed redistribution to the RBCs at all temperatures but only after low pH-induced fusion. The results are consistent with differential dispersion of viral components into the RBC and the existence of relatively long-lived barriers to diffusion subsequent to fusion pore formation.

  17. In vitro cytotoxicity, apoptosis, DNA-binding, and antioxidant activity studies of ruthenium (II) complexes.

    Science.gov (United States)

    Huang, Hong-Liang; Liu, Yun-Jun; Zeng, Cheng-Hui; He, Li-Xin; Wu, Fu-Hai

    2010-05-01

    Two new ligands maip (1) (maip = 2-(3-aminophenyl)imizado[4,5-f][1,10]phenanthroline), paip (2) (paip = 2-(4-aminophenyl)imidazo[4,5-f][1,10]phenanthroline), and their ruthenium (II) complexes [Ru(phen)(2)(maip)](ClO(4))(2) (3) and [Ru(phen)(2)(paip)](ClO(4))(2) (4) (phen = 1,10-phenanthroline) have been synthesized and characterized. The cytotoxicity of these compounds was evaluated by MTT [3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide] assay. The apoptosis assay was carried out with acridine orange/ethidium bromide staining methods. The DNA-binding behaviors of complexes 3 and 4 were investigated by viscosity measurements, thermal denaturation, photocleavage, and spectroscopic methods. The results show that the two complexes intercalate into the base pairs of DNA. In the presence of a complex, apoptosis of BEL-7402 cells was observed. Experiments show that these compounds exhibit antioxidant activity against hydroxyl radicals. PMID:20307189

  18. Is biofilm removal properly assessed? Comparison of different quantification methods in a 96-well plate system.

    Science.gov (United States)

    Stiefel, Philipp; Rosenberg, Urs; Schneider, Jana; Mauerhofer, Stefan; Maniura-Weber, Katharina; Ren, Qun

    2016-05-01

    Various methods have been reported to quantify total biofilm or different components of biofilm; however, these methods are often confusedly used, leading to discrepancies and misleading results. In this study, different methods for quantification of biofilm, including those for total biomass, total amount of bacterial cells, viable cell number, and amount of extracellular polymeric substances, were systematically compared in microtiter plates. To evaluate which method is suitable for assessment of biofilm removal and for bacterial killing, biofilm samples were treated with various cleaners possessing removing and/or killing capacities. It was found that most of the methods tested in this study in general exhibited high reproducibility and repeatability. Crystal Violet staining was a simple but reliable method for total biomass quantification. Total bacteria cell numbers could be reliably quantified by the fluorescent DNA-binding dye Acridine Orange. Viable cells could be quantified by either an ATP-based assay or a proliferation assay. Both of these viability methods showed a broad detection range and led to precise measurement. For quantification of proteins in the biofilm, staining with fluorescein isothiocyanate was most suitable. Furthermore, it was revealed that a combination of different methods is required to determine if a cleaner kills or removes biofilm. PMID:26923144

  19. Cellular heredity in haploid cultures of somatic cells. Annual progress report, March 1, 1975--March 31, 1976. [UV radiation

    Energy Technology Data Exchange (ETDEWEB)

    Freed, J.J.

    1976-01-01

    In experiments with haploid and diploid derivatives from the haploid frog embryo cell line ICR 2A, we have investigated aspects of cell survival, DNA repair and mutant induction after exposure to 254 nm radiation. Survival curves for haploid and diploid cells in random growth or blocked in the Gl phase of the cell cycle were determined; the survival data do not differ sufficiently to permit the use of such comparisons as an index of recessive lethal induction. Studies of the induction of thymine dimers in DNA indicated that the incidence of dimers in DNA from haploid and diploid cells is similar after exposure of the cells to equal doses of ultraviolet. The cells are capable of photoreversing dimers but appear to be deficient in excision repair. In an attempt to examine the effect of the permitted mode of DNA repair on the yield of mutations, we compared the incidence of ouabain-resistant variants among survivors of ultraviolet exposure and of ultraviolet exposure followed by photoreversal. Although the yield of resistant colonies was small, the data suggest that photoreversal lowers the yield of resistant colonies and thus that the induction of this phenotype is related to dimer persistence in DNA. We have also observed by fluorescence microscopy that an acridine mustard mutagen, ICR 191, is preferentially accumulated in cytoplasmic granules having the intracellular distribution pattern of lysosomes. This form of incorporation may be significant in the apparently non-genetic early toxicity of this compound observed in experiments with cultured cells.

  20. Enhanced accumulation of curcumin and temozolomide loaded magnetic nanoparticles executes profound cytotoxic effect in glioblastoma spheroid model.

    Science.gov (United States)

    Dilnawaz, Fahima; Sahoo, Sanjeeb Kumar

    2013-11-01

    Glioblastomas (GBMs) are highly lethal primary brain tumours. Treatment of these malignant gliomas remains ineffective as these are extremely resistant to chemotherapeutic applications. Furthermore, combination therapy for cancer treatment is becoming more popular because it generates synergistic anticancer effects, by reducing individual drug-related toxicity and associated side effects. Currently, magnetic nanoparticles (MNPs) based drug delivery system has attracted much more attention owing to its intrinsic magnetic properties and drug loading capacity. In the present study, MNPs based drug delivery approach for co-delivering of potent chemotherapeutic drugs such as Curcumin (herbal drug) and Temozolomide (DNA methylating agent) has been implemented. The dual drug loaded MNPs formulations were evaluated in two-dimensional (2-D) monolayer culture and three-dimensional (3-D) tumour spheroid culture of T-98G cells for understanding the therapeutic discrepancy. The dual drug loaded MNPs formulations demonstrated higher cytotoxic effect than single drug loaded MNPs formulations as compared to their corresponding native drugs in 2-D and 3-D culture. The combination index (CI) analysis revealed synergistic mode of action of dual drug loaded MNPs formulations, which was further confirmed by cell death induction assay mediated by acridine orange (AO)/propidium iodide (PI) staining, illustrating higher efficacy of the formulation towards GBM therapy.

  1. Calculating the contribution of different binding modes to Quinacrine - DNA complex formation from polarized fluorescence data

    CERN Document Server

    Voloshin, Igor; Karachevtsev, Victor; Zozulya, Victor

    2013-01-01

    Binding of acridine derivative quinacrine (QA) to chicken erythrocyte DNA was studied by methods of absorption and polarized fluorescent spectroscopy. Measurements were carried out in aqueous buffered solutions (pH 6.9) of different dye concentrations (QA concentration range from $10^{-6}$ till $10^{-4}$ M) and ionic strengths ($Na^{+}$ concentration rang from $10^{-3}$ till 0.15 M) in a wide range of phosphate-to-dye molar ratios ($P/D$). It is established that the minimum of fluorescent titration curve plotted as relative fluorescence intensity $vs$ $P/D$ is conditioned by the competition between the two types of QA binding to DNA which posses by different emission parameters: (i) intercalative one dominating under high $P/D$ values, and (ii) outside electrostatic binding dominating under low $P/D$ values, which is accompanied by the formation of non-fluorescent dye associates on the DNA backbone. Absorption and fluorescent characteristics of complexes formed were determined. The method of calculation of di...

  2. Isolation of a flavonoid, apigenin 7-O-glucoside, from Mentha longifolia (L.) Hudson subspecies longifolia and its genotoxic potency.

    Science.gov (United States)

    Gulluce, Medine; Orhan, Furkan; Yanmis, Derya; Arasoglu, Tulin; Guvenalp, Zuhal; Demirezer, Lutfiye Omur

    2015-09-01

    Mentha is a medicinal and aromatic plant belonging to the Lamiaceae family, which is widely used in food, flavor, cosmetic and pharmaceutical industries. Recently, it has been found that the use of Mentha as a pharmaceutical source is based on its phytochemical constituents that have far been identified as tannins, saponins, phenolic acids and flavonoids. This study was designed to evaluate the mutagenic and antimutagenic activities of apigenin 7-O-glucoside (A7G), a flavonoid isolated from Mentha longifolia (L.) Hudson subspecies longifolia (ML). The possible antimutagenic potential of A7G was examined against mutagens ethyl methanesulfonate and acridine in an eukaryotic cell system Saccharomyces cerevisiae and sodium azide in Salmonella typhimurium TA1535 and 9-aminoacridine in S. typhimurium TA1537. According to our findings, any concentrations of the A7G used did not show mutagenic activity but exerted strong antimutagenic activities at tested concentrations. The inhibition rates for the Ames test ranged from 27.2% (S. typhimurium TA1535: 0.4 μM/plate) to 91.1% (S. typhimurium TA1537: 0.2 μM/plate) and for the yeast deletion assay from 4% to 57.7%. This genotoxicological study suggests that a flavonoid from ML owing to antimutagenic properties is of great pharmacological importance and might be beneficial to industries producing food additives, cosmetics and pharmaceuticals products.

  3. Studies on Infectious Mononucleosis

    Science.gov (United States)

    Joncas, J.; Chagnon, A.; Pavilanis, V.

    1966-01-01

    Viral studies were carried out on throat swabs, rectal swabs and washed white blood cells from 27 cases of infectious mononucleosis (positive Paul-Bunnell-David-sohn test), and from 22 controls. Four cytopathic agents were isolated in the test group, two of which were readily subcultured for at least three successive passages. Three cytopathic agents were recovered in the control group, two of which have been identified as adenovirus type 5 and adenovirus type 3. The unidentified agents tested so far are sensitive to ether and to pH 3. The results of acridine-orange staining and the immunofluorescence technique, using a conjugated control serum and two conjugated convalescent infectious mononucleosis sera, indicate that the isolated agent or agents in the test group are RNA-type agents with a cytoplasmic cycle of development. The overall results of this study lead the authors to suspect a respiratory syncytial-like myxovirus as the as yet unidentified agent which they recovered. ImagesFig. 1aFig. 1bFig. 1cFig. 1dFig. 2aFig. 2bFig. 2cFig. 2dFig. 3aFig. 3bFig. 3cFig. 3dFig. 3eFig. 3f PMID:4952899

  4. Synthesis and biological evaluation of oxindole linked indolyl-pyrimidine derivatives as potential cytotoxic agents.

    Science.gov (United States)

    Prajapti, Santosh Kumar; Nagarsenkar, Atulya; Guggilapu, Sravanthi Devi; Gupta, Keshav Kumar; Allakonda, Lingesh; Jeengar, Manish Kumar; Naidu, V G M; Babu, Bathini Nagendra

    2016-07-01

    In our endeavor towards the development of effective cytotoxic agents, a series of oxindole linked indolyl-pyrimidine derivatives were synthesized and characterized by IR, (1)H NMR, (13)C NMR and Mass spectral analysis. All the newly synthesized target compounds were assessed against PA-1 (ovarian), U-87MG (glioblastoma), LnCaP (prostate), and MCF-7 (Breast) cancer cell lines for their cytotoxic potential, with majority of them showing inhibitory activity at low micro-molar concentrations. Significantly, compound 8e was found to be most potent amongst all the tested compounds with an IC50 value of (2.43±0.29μM) on PA-1 cells. The influence of the most active cytotoxic compound 8e on the cell cycle distribution was assessed on the PA-1 cell line, exhibiting a cell cycle arrest at the G2/M phase. Moreover, acridine orange/ethidium bromide staining and annexin V binding assay confirmed that compound 8e can induce cell apoptosis in PA-1 cells. These preliminary results persuade further investigation on the synthesized compounds aiming to the development of potential cytotoxic agents.

  5. Cytotoxic evaluation of different fractions of Salvia chorassanica Bunge on MCF-7 and DU 145 cell lines.

    Science.gov (United States)

    Golshan, Alireza; Amini, Elaheh; Emami, Seyed Ahmad; Asili, Javad; Jalali, Zahra; Sabouri-Rad, Sarvenaz; Sanjar-Mousavi, Naghmeh; Tayarani-Najaran, Zahra

    2016-01-01

    Because of antimicrobial, antioxidant, and anticancer potential, Salvia chorassanica Bunge (Lamiaceae) has been considered as a popular herb in Iranian traditional medicine. Previous studies have shown remarkable cytotoxic properties of the methanol, n-hexane and dichloromethane extract of S. chorassanica on human cervical cancer cells. To seek the therapeutic potentials of S. chorassanica, this study was undertaken to evaluate the cytotoxic activities of various extracts of this plant on human breast MCF-7 and prostate cancer DU 145 cells. The DU 145 cells were exposed to different concentrations of plant extracts (1-200 μg/ml). Cytotoxic activities were examined using alamarBlue(®) assay and apoptosis was assessed by acridine orange/propodium iodide double staining and evaluation of DNA fragmentation by flow cytometry. Our findings indicated that n-hexane and dichloromethane extracts had more cytotoxic activities against DU 145 and MCF-7 cell lines compared with other extracts (Pcytotoxic effects of S. chorassanica extracts on MCF-7 and DU 145 cell lines which is most likely exerted via apoptosis cell death. Therefore, further investigations on S. chorassanica extracts as potential chemotherapeutic agents are warranted.

  6. Pyridinoacridine alkaloids of marine origin: NMR and MS spectral data, synthesis, biosynthesis and biological activity

    Directory of Open Access Journals (Sweden)

    Louis P. Sandjo

    2015-09-01

    Full Text Available This review focuses on pyridoacridine-related metabolites as one biologically interesting group of alkaloids identified from marine sources. They are produced by marine sponges, ascidians and tunicates, and they are structurally comprised of four to eight fused rings including heterocycles. Acridine, acridone, dihydroacridine, and quinolone cores are features regularly found in these alkaloid skeletons. The lack of hydrogen atoms next to quaternary carbon atoms for two or three rings makes the chemical shift assignment a difficult task. In this regard, one of the aims of this review is the compilation of previously reported, pyridoacridine 13C NMR data. Observations have been made on the delocalization of electrons and the presence of some functional groups that lead to changes in the chemical shift of some carbon resonances. The lack of mass spectra information for these alkaloids due to the compactness of their structures is further discussed. Moreover, the biosynthetic pathways of some of these metabolites have been shown since they could inspire biomimetic synthesis. The synthesis routes used to prepare members of these marine alkaloids (as well as their analogues, which are synthesized for biological purposes are also discussed. Pyridoacridines were found to have a large spectrum of bioactivity and this review highlights and compares the pharmacophores that are responsible for the observed bioactivity.

  7. Fuzzy logic sensing of G-quadruplex DNA and its cleavage reagents based on reduced graphene oxide.

    Science.gov (United States)

    Huang, Wei Tao; Zhang, Jian Rong; Xie, Wan Yi; Shi, Yan; Luo, Hong Qun; Li, Nian Bing

    2014-07-15

    Herein, by combining the merits of nanotechnology and fuzzy logic theory, we develop a simple, label-free, and general strategy based on an organic dye-graphene hybrid system for fluorescence intelligent sensing of G-quadruplexes (G4) formation, hydroxyl radical (HO∙), and Fe(2+) in vitro. By exploiting acridine orange (AO) dyes-graphene as a nanofilter and nanoswitch and the ability of graphene to interact with DNA with different structures, our approach can efficiently distinguish, quantitatively detect target analytes. In vitro assays with G4DNA demonstrated increases in fluorescence intensity of the AO-rGO system with a linear range of 16-338 nM and a detection limit as low as 2.0 nM. The requenched fluorescence of the G4TBA-AO-rGO system has a non-linear response to Fenton reagent. But this requenching reduces the fluorescence intensity in a manner proportional to the logarithm to the base 10 of the concentration of Fenton reagent in the range of 0.1-100 μM and 100-2000 μM, respectively. Furthermore, we develop a novel and intelligent sensing method based on fuzzy logic which mimics human reasoning, solves complex and non-linear problems, and transforms the numerical output into the language description output for potential application in biochemical systems, environmental monitoring systems, and molecular-level fuzzy logic computing system.

  8. Host–guest composite materials of dyes loaded zeolite LTL for antenna applications

    International Nuclear Information System (INIS)

    This research work directly focuses on a new feasible light harvesting antenna material constructed with Acridine hydrochloride (Ac)/Acriflavine hydrochloride (AF), as donor/acceptor for energy transfer, loaded on a round shape zeolite LTL (K-LTL and H-LTL). The energy transfer was monitored by absorption and fluorescence spectra while the calculated Förster distance (RDA) and Quenching efficiency (%Q) of Ac/AF on K-LTL and H-LTL varied between 22.0 Å to 19.6 Å and 71.4% to 65.5%, respectively. Also, it was found that the microenvironment of a solid host such as K-LTL and H-LTL has significantly influenced the fluorescence spectra of Ac/AF on H-LTL approximately 50 nm longer than that on K-LTL. - Highlights: • New antenna materials have been performed using dyes loaded on zeolite LTL. • Light emission takes place from acriflavine hydrochloride (AF) due to fluorescence resonance energy transfer (FRET). • The microenvironment of zeolite LTL has significantly influenced the fluorescence spectra

  9. Bacterial communities in the fruit bodies of ground basidiomycetes

    Science.gov (United States)

    Zagryadskaya, Yu. A.; Lysak, L. V.; Chernov, I. Yu.

    2015-06-01

    Fruit bodies of basidiomycetes at different stages of decomposition serve as specific habitats in forest biocenoses for bacteria and differ significantly with respect to the total bacterial population and abundance of particular bacterial genera. A significant increase in the total bacterial population estimated by the direct microscopic method with acridine orange staining and in the population of saprotrophic bacteria (inoculation of glucose peptone yeast agar) in fruit bodies of basidiomycetes Armillaria mellea and Coprinus comatus was recorded at the final stage of their decomposition in comparison with the initial stage. Gramnegative bacteria predominated in the tissues of fruit bodies at all the stages of decomposition and were represented at the final stage by the Aeromonas, Vibrio, and Pseudomonas genera (for fruit bodies of A. mellea) the Pseudomonas genus (for fruit bodies of C. comatus). The potential influence of bacterial communities in the fruit bodies of soil basidiomycetes on the formation of bacterial communities in the upper soil horizons in forest biocenoses is discussed. The loci connected with the development and decomposition of fruit bodies of basidiomycetes on the soil surface are promising for targeted search of Gram-negative bacteria, the important objects of biotechnology.

  10. Hesperidin as a preventive resistance agent in MCF-7 breast cancer cells line resistance to doxorubicin

    Institute of Scientific and Technical Information of China (English)

    Rifki Febriansah; Dyaningtyas Dewi PP; Sarmoko; Nunuk Aries Nurulita; Edy Meiyanto; Agung Endro Nugroho

    2014-01-01

    Objective:To evaluate of hesperidin to overcome resistance of doxorubicin in MCF-7 resistant doxorubicin cells (MCF-7/Dox) in cytotoxicity apoptosis and P-glycoprotein (Pgp) expression in combination with doxorubicin. Methods:The cytotoxic properties, 50%inhibition concentration (IC50) and its combination with doxorubicin in MCF-7 cell lines resistant to doxorubicin (MCF-7/Dox) cells were determined using MTT assay. Apoptosis induction was examined by double staining assay using ethidium bromide-acridine orange. Immunocytochemistry assay was performed to determine the level and localization of Pgp. Results: Single treatment of hesperidin showed cytotoxic activity on MCF-7/Dox cells with IC50 value of 11 µmol/L. Thus, combination treatment from hesperidin and doxorubicin showed addictive and antagonist effect (CI>1.0). Hesperidin did not increase the apoptotic induction, but decreased the Pgp expressions level when combined with doxorubicin in low concentration. Conclusions: Hesperidin has cytotoxic effect on MCF-7/Dox cells with IC50 of 11 µmol/L. Hesperidin did not increased the apoptotic induction combined with doxorubicin. Co-chemotherapy application of doxorubicin and hesperidin on MCF-7/Dox cells showed synergism effect through inhibition of Pgp expression.

  11. ANTIPROLIFERATIVE AND APOPTOTIC EFFECTS OF THE ESSENTIAL OIL OF ORIGANUM ONITES AND CARVACROL ON HEP-G2 CELLS

    Directory of Open Access Journals (Sweden)

    Özlem TOMSUK

    2011-08-01

    Full Text Available The essential oil Origanum onites L. and its phenolic constituent carvacrol were examined for their cytotoxic and apoptotic effects in a human hepatocellular carcinoma cells Hep-G2. WST-1 and neutral red uptake assays were performed to determine the inhibitory effects of the oil and carvacrol on the growth of the cells. Possible induction of apoptosis by Origanum oil and carvacrol was further investigated by acridine orange/ethidium bromide (AO/EB staining. Results showed that the Ori- ganum oil and carvacrol was significantly cytotoxic and induced apoptosis in Hep-G2 cells. IC₅₀ value of essential oil and carvacrol was found about 0,009% (v/v and 500 μM, respectively. After incuba- tion of the cells with Origanum oil and carvacrol, characteristics of apoptotic morphology such as chromatin condensation, shrinkage of the cells and cytoplasmic blebbing was observed. In conclusion, both essential oil and its major constituent carvacrol significantly exhibited cytotoxic and apoptotic activities in hepatocellular carcinoma cells, indicating its potential for use as an anticancer agent.

  12. Combinational effects of hexane insoluble fraction of Ficus septica Burm. F. and doxorubicin chemotherapy on T47D breast cancer cells

    Institute of Scientific and Technical Information of China (English)

    Agung Endro Nugroho; Adam Hermawan; Anindya Novika; Edy Meiyanto

    2013-01-01

    Objective: To evaluate the effects of n-hexane insoluble fraction (HIF) of Ficus septica leaves in combination with doxorubicin on cytotoxicity, cell cycle and apoptosis induction of breast cancer T47D cell lines. Methods: The in vitro drugs-stimulated cytotoxic effects were determined using MTT assay. Analysis of cell cycle distribution was performed using flowcytometer and the data was analyzed using ModFit LT 3.0 program. Apoptosis assay was carried out by double staining method using ethydium bromide-acridin orange. The expression of cleaved-poly (ADP-ribose) polymerase (PARP) on T47D cell lines was identified using immunocytochemistry. Results:The combination exhibited higher inhibitory effect on cell growth than the single treatment of doxorubicin in T47D cells. In addition, combination of doxorubicin and HIF increased the incidence of cells undergoing apoptosis. HIF could improve doxorubicin cytotoxic effect by changing the accumulation of cell cycle phase from G2/M to G1 phase. The combination also exhibited upregulation of cleaved-PARP in T47D cells. Conclusions: Based on this results, HIF is potential to be developed as co-chemotherapeutic agent for breast cancer by inducing apoptosis and cell cycle arrest. However, the molecular mechanism need to be explored further.

  13. Destruction of Gastric Cancer Cells to Mesothelial Cells by Apoptosis in the Early Peritoneal Metastasis

    Institute of Scientific and Technical Information of China (English)

    Di NA; Funan LIU; Zhifeng MIAO; Zongmin DU; Huimian XU

    2009-01-01

    This study examined the mechanism by which the gastric cancer cells lead to early peritoneal metastasis.HMrSV5 cells,a human peritoneal mesothelial cell line,were co-incubated with the supernatants of gastric cancer cells.Morphological changes of HMrSV5 cells were observed.The cell damage was quantitatively determined by MTT assay.The apoptosis of HMrSV5 cells was observed under transmission electron microscope.Acridine orange/ethidium bromidestained condensed nuclei was detected by fluorescent microscopy and flow cytometry.The expressions of Bcl-2 and Bax was immunochemically evaluated.The results showed that conspicuous morphological changes of apoptosis were observed in HMrSV5 cells 24 h after treatment with the supematants of gastric cancer cells.The supernatants could induce apoptosis of HMrSV5 cells in a time-dependent manner.Th esupematants could up-regulate the expression of Bax and suppress that of Bcl-2 in HMrSV5 cells.These findings demonstrated that gastric cancer cells can induce the apoptosis of HPMCs through supernatants in the early peritoneal metastasis.The abnormal expressions of Bcl-2 and Bax may contribute to the apoptosis.Anti-apoptosis drugs promise to be adjuvant chemotherapeutic agents in the treatment of peritoneal metastasis of gastric cancer.

  14. Cytotoxicity of α-terpineol in HeLa cell line and its effects to apoptosis and cell cycle

    Directory of Open Access Journals (Sweden)

    Rasuane Noor Indwiani Astuti Mustofa

    2014-08-01

    Full Text Available α-Terpineol is a natural compound of terpenoid alcohols class. However, it can be synthesizedfrom α-pinene of turpentin content. α-Terpineol has been reported as potential anticancer agentdue to its activity on inhibition of cells growth and induction of tumor cell death. However, itsanticancer activity in HeLa cervical cancer cells line has never been studied, yet. The aim of thisstudy was to evaluate the cytotoxicity of α-terpineol and its effects to apoptosis and cell cycle.This was a quasi-experimental study with post-test only with non-equivalent control groupdesign. Cytotoxicity of á-terpineol was evaluated using MTT cell viability assay. The effect of α-terpineol on cell apoptotis was tested using acridine orange-ethidium bromide staining method,whereas its effect on cell cycle was evaluated by flowcytometry method. The results showedthat α-terpineol had cytotoxicity against HeLa cell with an IC50 value about 12.46 μg/mL.Furthermore, α-terpineol induced the HeLa with an IC50 value about 13.12 μg/mL. Cell accumulationat G1 phase during cell cycle after incubation with α-terpineol (52.78was observed. In conclusion,α-terpineol is potential as an anticancer due to its ability to induce cell apoptosis and to inhibitthe cell cycle at G1 phase.

  15. Caffeine-Induced Premature Chromosome Condensation Results in the Apoptosis-Like Programmed Cell Death in Root Meristems of Vicia faba.

    Directory of Open Access Journals (Sweden)

    Dorota Rybaczek

    Full Text Available We have demonstrated that the activation of apoptosis-like programmed cell death (AL-PCD was a secondary result of caffeine (CF induced premature chromosome condensation (PCC in hydroxyurea-synchronized Vicia faba root meristem cells. Initiation of the apoptotic-like cell degradation pathway seemed to be the result of DNA damage generated by treatment with hydroxyurea (HU [double-stranded breaks (DSBs mostly] and co-treatment with HU/CF [single-stranded breaks (SSBs mainly]. A single chromosome comet assay was successfully used to study different types of DNA damage (neutral variant-DSBs versus alkaline-DSBs or SSBs. The immunocytochemical detection of H2AXS139Ph and PARP-2 were used as markers for DSBs and SSBs, respectively. Acridine orange and ethidium bromide (AO/EB were applied for quantitative immunofluorescence measurements of dead, dying and living cells. Apoptotic-type DNA fragmentation and positive TUNEL reaction finally proved that CF triggers AL-PCD in stressed V. faba root meristem cells. In addition, the results obtained under transmission electron microscopy (TEM further revealed apoptotic-like features at the ultrastructural level of PCC-type cells: (i extensive vacuolization; (ii abnormal chromatin condensation, its marginalization and concomitant degradation; (iii formation of autophagy-like vesicles (iv protoplast shrinkage (v fragmentation of cell nuclei and (vi extensive degeneration of the cells. The results obtained have been discussed with respect to the vacuolar/autolytic type of plant-specific AL-PCD.

  16. Antiproliferative and Proapoptotic Activities of Methanolic Extracts from Ligustrum vulgare L. as an Individual Treatment and in Combination with Palladium Complex

    Directory of Open Access Journals (Sweden)

    Snežana D. Marković

    2012-02-01

    Full Text Available The aim of this study is to examine the growth inhibitory effects of methanolic leaf and fruit extracts of L. vulgare on HCT-116 cells over different time periods and their synergistic effect with a Pd(apox complex. The antiproliferative activity of plant extracts alone or in combination with the Pd(apox complex was determined using MTT cell viability assay, where the IC50 value was used as a parameter of cytotoxicity. Results show that antiproliferative effects of L. vulgare extracts increase with extension of exposure time, with decreasing IC50 values, except for 72 h where the IC50 values for methanolic leaf extract were lower than for the fruit extract. The Pd(apox complex alone had a weak antiproliferative effect, but combination with L. vulgare extracts caused stronger effects with lower IC50 values than with L. vulgare extracts alone. The type of cell death was explored by fluorescence microscopy using the acridin orange/ethidium bromide method. Treatments with plant extracts caused typical apoptotic morphological changes in HCT-116 cells and co-treatments with Pd(apox complex caused higher levels of apoptotic cells than treatment with plant extracts alone. The results indicate that L. vulgare is a considerable source of natural bioactive substances with antiproliferative activity on HCT-116 cells and which have a substantial synergistic effect with the Pd(apox complex.

  17. Chemotactic behavior of deep subsurface bacteria toward carbohydrates, amino acids and a chlorinated alkene

    Energy Technology Data Exchange (ETDEWEB)

    Lopez de Victoria, G. (Puerto Rico Univ., Rio Piedras (Puerto Rico). Dept. of Biology)

    1989-02-01

    The chemotactic behavior of deep terrestrial subsurface bacteria toward amino acids, carbohydrates and trichloroethylene was assayed using a modification of the capillary method and bacterial enumeration by acridine orange direct counts. Eleven isolates of bacteria isolated from six different geological formations were investigated. A bimodal response rather than an absolute positive or negative response was observed in most assays. Most of the isolates were positively chemotactic to low concentrations of substrates and were repelled by high concentrations of the same substrate. However, this was not the case for trichloroethylene (TCE) which was mostly an attractant and elicited the highest responses in all the isolates when compared with amino acids and carbohydrates. The movement rates of these isolates in aseptic subsurface sediments in the absence and presence of TCE were also determined using a laboratory model. All of the isolates showed distinct response range, peak, and threshold concentrations when exposed to the same substrates suggesting that they are possibly different species as has been inferred from DNA homology studies. 101 refs., 4 figs., 57 tabs.

  18. Transient absorption spectroscopy in biology using the Super-ACO storage ring FEL and the synchrotron radiation combination

    CERN Document Server

    Renault, E; De Ninno, G; Garzella, D; Hirsch, M; Nahon, L; Nutarelli, D

    2001-01-01

    The Super-ACO storage ring FEL, covering the UV range down to 300 nm with a high average power (300 mW at 350 nm) together with a high stability and long lifetime, is a unique tool for the performance of users applications. We present here the first pump-probe two color experiments on biological species using a storage ring FEL coupled to the synchrotron radiation. The intense UV pulse of the Super-ACO FEL is used to prepare a high initial concentration of chromophores in their first singlet electronic excited state. The nearby bending magnet synchrotron radiation provides, on the other hand a pulsed, white light continuum (UV-IR), naturally synchronized with the FEL pulses and used to probe the photochemical subsequent events and the associated transient species. We have demonstrated the feasibility with a dye molecule (POPOP) observing a two-color effect, signature of excited state absorption and a temporal signature with Acridine. Applications on various chromophores of biological interest are carried out,...

  19. Development of an analytical method for the simultaneous determination of 15 carcinogenic polycyclic aromatic hydrocarbons and polycyclic aromatic nitrogen heterocyclic compounds. application to diesel particulates.

    Science.gov (United States)

    Sauvain, J J; Vu Duc, T; Huynh, C K

    2001-12-01

    A new method enabling the determination of 15 priority carcinogenic polyaromatic compounds (PAC) proposed by the US National Toxicology Program (NTP) has been developed and applied to diesel exhaust particulates (DEP). The clean-up procedure consists of solid-phase extraction (SPE) and HPLC fractionation on silica phases followed by liquid-liquid extraction and chromatography on a polyvinylbenzene copolymer column. The method gives good recoveries for all PAC studied except dibenzo[a,j]acridine and dibenzo[a,h]pyrene, for which recovery values are below 80%. The use of GC-MS ion trap and its capacity to achieve single-ion storage enhanced the sensitivity of the method, enabling the detection of high-molecular-weight PAH in the low ng g(-1) concentration range. Intermediate polarity GC columns, e.g. BPX-50 or equivalent, enabled better separation, when applied to DEP analysis, than the generally used DB-5 apolar phase. This is observed mainly for separation of isomeric compounds belonging to the benzofluoranthene and dibenzopyrene families. The application of this method to DEP sampled from the exhaust of a diesel engine and in confined locations such as a tunnel has shown that all PAH of the NTP list could be detected, except dibenzo[a,h]pyrene. No dibenzacridine or dibenzocarbazole could be detected in such matrices. The method is sufficiently sensitive to be applicable to environmental exposure measurements in occupational health surveys.

  20. Immunological techniques as tools to characterize the subsurface microbial community at a trichloroethylene contaminated site

    Energy Technology Data Exchange (ETDEWEB)

    Fliermans, C.B.; Dougherty, J.M.; Franck, M.M.; McKinzey, P.C.; Hazen, T.C.

    1992-01-01

    Effective in situ bioremediation strategies require an understanding of the effects pollutants and remediation techniques have on subsurface microbial communities. Therefore, detailed characterization of a site's microbial communities is important. Subsurface sediment borings and water samples were collected from a trichloroethylene (TCE) contaminated site, before and after horizontal well in situ air stripping and bioventing, as well as during methane injection for stimulation of methane-utilizing microorganisms. Subsamples were processed for heterotrophic plate counts, acridine orange direct counts (AODC), community diversity, direct fluorescent antibodies (DFA) enumeration for several nitrogen-transforming bacteria, and Biolog [reg sign] evaluation of enzyme activity in collected water samples. Plate counts were higher in near-surface depths than in the vadose zone sediment samples. During the in situ air stripping and bioventing, counts increased at or near the saturated zone, remained elevated throughout the aquifer, but did not change significantly after the air stripping. Sporadic increases in plate counts at different depths as well as increased diversity appeared to be linked to differing lithologies. AODCs were orders of magnitude higher than plate counts and remained relatively constant with depth except for slight increases near the surface depths and the capillary fringe. Nitrogen-transforming bacteria, as measured by serospecific DFA, were greatly affected both by the in situ air stripping and the methane injection. Biolog[reg sign] activity appeared to increase with subsurface stimulation both by air and methane. The complexity of subsurface systems makes the use of selective monitoring tools imperative.

  1. Immunological techniques as tools to characterize the subsurface microbial community at a trichloroethylene contaminated site

    Energy Technology Data Exchange (ETDEWEB)

    Fliermans, C.B.; Dougherty, J.M.; Franck, M.M.; McKinzey, P.C.; Hazen, T.C.

    1992-12-31

    Effective in situ bioremediation strategies require an understanding of the effects pollutants and remediation techniques have on subsurface microbial communities. Therefore, detailed characterization of a site`s microbial communities is important. Subsurface sediment borings and water samples were collected from a trichloroethylene (TCE) contaminated site, before and after horizontal well in situ air stripping and bioventing, as well as during methane injection for stimulation of methane-utilizing microorganisms. Subsamples were processed for heterotrophic plate counts, acridine orange direct counts (AODC), community diversity, direct fluorescent antibodies (DFA) enumeration for several nitrogen-transforming bacteria, and Biolog {reg_sign} evaluation of enzyme activity in collected water samples. Plate counts were higher in near-surface depths than in the vadose zone sediment samples. During the in situ air stripping and bioventing, counts increased at or near the saturated zone, remained elevated throughout the aquifer, but did not change significantly after the air stripping. Sporadic increases in plate counts at different depths as well as increased diversity appeared to be linked to differing lithologies. AODCs were orders of magnitude higher than plate counts and remained relatively constant with depth except for slight increases near the surface depths and the capillary fringe. Nitrogen-transforming bacteria, as measured by serospecific DFA, were greatly affected both by the in situ air stripping and the methane injection. Biolog{reg_sign} activity appeared to increase with subsurface stimulation both by air and methane. The complexity of subsurface systems makes the use of selective monitoring tools imperative.

  2. Effect of storage in short--and long-term commercial semen extenders on the motility, plasma membrane and chromatin integrity of boar spermatozoa.

    Science.gov (United States)

    De Ambrogi, Marco; Ballester, Juan; Saravia, Fernando; Caballero, Ignacio; Johannisson, Anders; Wallgren, Margareta; Andersson, Magnus; Rodriguez-Martinez, Heriberto

    2006-10-01

    For artificial insemination (AI) in pigs, preservation of liquid boar semen at 16-20 degrees C is still common practice as sperm cryopreservation remains suboptimal in this species. To meet the different needs of the swine industry, several extenders have been developed to preserve semen in liquid form for short--and long-term storage. In the present study, three different commercial extenders devised for short-term (BTS+) or long-term preservation (MR-A and X-Cell), were used to test whether storage of semen from four mature, fertile boars at 17 degrees C for 96 h would affect sperm characteristics relevant for fertility, such as motility, membrane integrity and chromatin stability. Computer-assisted sperm analysis, and stainings with the acylated membrane dye SYBR-14/propidium iodide, and acridine orange in connection with flow cytometry were used to evaluate these variables. Percentages of total motile spermatozoa decreased slightly, but significantly, after 72-96 h. While membrane integrity values varied during the period of study, no significant changes in either membrane integrity or chromatin stability were, however, registered. This suggests a customary 96-day storage at 17 degrees C in these extenders was too short an interval to cause losses of integrity in nuclear DNA in the boar population studied. PMID:16573706

  3. Imaging and measuring the rituximab-induced changes of mechanical properties in B-lymphoma cells using atomic force microscopy

    Energy Technology Data Exchange (ETDEWEB)

    Li, Mi [State Key Laboratory of Robotics, Shenyang Institute of Automation, Chinese Academy of Sciences, Shenyang 110016 (China); Graduate University of Chinese Academy of Sciences, Beijing 100049 (China); Liu, Lianqing, E-mail: lqliu@sia.cn [State Key Laboratory of Robotics, Shenyang Institute of Automation, Chinese Academy of Sciences, Shenyang 110016 (China); Xi, Ning, E-mail: xin@egr.msu.edu [State Key Laboratory of Robotics, Shenyang Institute of Automation, Chinese Academy of Sciences, Shenyang 110016 (China); Department of Electrical and Computer Engineering, Michigan State University, East Lansing, MI 48824 (United States); Wang, Yuechao; Dong, Zaili [State Key Laboratory of Robotics, Shenyang Institute of Automation, Chinese Academy of Sciences, Shenyang 110016 (China); Tabata, Osamu [Department of Micro Engineering, Kyoto University, Kyoto 606-8501 (Japan); Xiao, Xiubin [Department of Lymphoma, Affiliated Hospital of Military Medical Academy of Sciences, Beijing 100071 (China); Zhang, Weijing, E-mail: zhangwj3072@163.com [Department of Lymphoma, Affiliated Hospital of Military Medical Academy of Sciences, Beijing 100071 (China)

    2011-01-14

    Research highlights: {yields} Single B-lymphoma living cells were imaged by AFM with the assistance of microfabricated pillars. {yields} The apoptosis of B-lymphoma cells triggered by rituximab without cross-linking was observed by AO/EB double fluorescent staining. {yields} The B-lymphoma cells became dramatically softer after adding rituximab. -- Abstract: The topography and mechanical properties of single B-lymphoma cells have been investigated by atomic force microscopy (AFM). With the assistance of microfabricated patterned pillars, the surface topography and ultrastructure of single living B-lymphoma cell were visualized by AFM. The apoptosis of B-lymphoma cells induced by rituximab alone was observed by acridine orange/ethidium bromide (AO/EB) double fluorescent staining. The rituximab-induced changes of mechanical properties in B-lymphoma cells were measured dynamically and the results showed that B-lymphoma cells became dramatically softer after incubation with rituximab. These results can improve our understanding of rituximab'effect and will facilitate the further investigation of the underlying mechanisms.

  4. Hydrogen sulfide lowers proliferation and induces protective autophagy in colon epithelial cells.

    Directory of Open Access Journals (Sweden)

    Ya C Wu

    Full Text Available Hydrogen sulfide (H(2S is a gaseous bacterial metabolite that reaches high levels in the large intestine. In the present study, the effect of H(2S on the proliferation of normal and cancerous colon epithelial cells was investigated. An immortalized colon epithelial cell line (YAMC and a panel of colon cancer cell lines (HT-29, SW1116, HCT116 were exposed to H(2S at concentrations similar to those found in the human colon. H(2S inhibited normal and cancerous colon epithelial cell proliferation as measured by MTT assay. The anti-mitogenic effect of H(2S was accompanied by G(1-phase cell cycle arrest and the induction of the cyclin-dependent kinase inhibitor p21(Cip. Moreover, exposure to H(2S led to features characteristic of autophagy, including increased formation of LC3B(+ autophagic vacuoles and acidic vesicular organelles as determined by immunofluorescence and acridine orange staining, respectively. Abolition of autophagy by RNA interference targeting Vps34 or Atg7 enhanced the anti-proliferative effect of H(2S. Further mechanistic investigation revealed that H(2S stimulated the phosphorylation of AMP-activated protein kinase (AMPK and inhibited the phosphorylation of mammalian target of rapamycin (mTOR and S6 kinase. Inhibition of AMPK significantly reversed H(2S-induced autophagy and inhibition of cell proliferation. Collectively, we demonstrate that H(2S inhibits colon epithelial cell proliferation and induces protective autophagy via the AMPK pathway.

  5. Bees' subtle colour preferences: how bees respond to small changes in pigment concentration

    Science.gov (United States)

    Papiorek, Sarah; Rohde, Katja; Lunau, Klaus

    2013-07-01

    Variability in flower colour of animal-pollinated plants is common and caused, inter alia, by inter-individual differences in pigment concentrations. If and how pollinators, especially bees, respond to these small differences in pigment concentration is not known, but it is likely that flower colour variability impacts the choice behaviour of all flower visitors that exhibit innate and learned colour preferences. In behavioural experiments, we simulated varying pigment concentrations and studied its impact on the colour choices of bumblebees and honeybees. Individual bees were trained to artificial flowers having a specific concentration of a pigment, i.e. Acridine Orange or Aniline Blue, and then given the simultaneous choice between three test colours including the training colour, one colour of lower and one colour of higher pigment concentration. For each pigment, two set-ups were provided, covering the range of low to middle and the range of middle to high pigment concentrations. Despite the small bee-subjective perceptual contrasts between the tested stimuli and regardless of training towards medium concentrations, bees preferred neither the training stimuli nor the stimuli offering the highest pigment concentration but more often chose those stimuli offering the highest spectral purity and the highest chromatic contrast against the background. Overall, this study suggests that bees choose an intermediate pigment concentration due to its optimal conspicuousness. It is concluded that the spontaneous preferences of bees for flower colours of high spectral purity might exert selective pressure on the evolution of floral colours and of flower pigmentation.

  6. [Effect of osthol on apoptosis and bone resorption of osteoclasts cultured in vitro].

    Science.gov (United States)

    Ming, Lei-Guo; Wang, Ming-Gang; Chen, Ke-Ming; Zhou, Jian; Han, Gui-Qiu; Zhu, Rui-Qing

    2012-02-01

    This study is to investigate the effect of osthol on osteoclasts' activity, bone resorption as well as apoptosis in vitro, and explore the mechanism of osthol in preventing osteoporosis. Osteoclasts were separated from long-limb bones of new born rabbits, cultured in 24-well plate with glass slices and bone slices, and treated by 1 x 10(-5) mol x L(-1) osthol. Osteoclasts were identified by observing live cells with phase contrast microscope, HE staining, TRAP staining and toluidine blue staining of bone resorption pits. The numbers of bone resorption pits were counted as well as the surface area of bone resorption on bone slice. Osteoclasts were stained with acridine orange to detect the cell apoptosis. The ratio of apoptotic osteoclasts was observed under fluorescence microscope. The gene expression of RANKL, OPG, TRAP and p-JNK1/2 protein expression were examined using real time PCR and Western blotting, respectively. Comparing with the control group without osthol, the rates of apoptotic osteoclasts increased obviously and the number and area of bone resorption pits decreased evidently with 1 x 10(-5) mol x L(-1) osthol. There is significant difference between control group and experiment group treated by 1 x 10(-5) mol x L(-1) osthol. Therefore, the osthol through RANK+RANKL/TRAF6/Mkk/JNK signal pathway inhibits the osteoclasts activity, enhances osteoclasts apoptotic and inhibits the bone resorption. PMID:22512027

  7. High-LET radiation enhanced apoptosis but not necrosis regardless of p53 status

    International Nuclear Information System (INIS)

    Purpose: We analyzed the death pattern of human lung cancer cells harboring different p53 statuses after irradiation with different levels of linear energy transfer (LET). Methods and materials: We used three kinds of human lung cancer cell lines with identical genotypes, except for the p53 gene. These cells were exposed to X-rays or accelerated carbon-ion beams. The cellular sensitivities were determined by a colony-forming assay. The detection and quantification of cell death (apoptosis and necrosis) were evaluated and compared by acridine orange/ethidium bromide double staining for fluorescence microscopy. Results: We found that (1) there was no significant difference in cellular sensitivity to LET radiation >70 KeV/μm, although wild-type p53 cell sensitivity to X-rays was higher than that of mutated p53 or p53-null cells; (2) low-LET radiation effectively induced apoptosis in wild-type p53 cells as compared with mutated p53 and p53-null cells; and (3) high-LET radiation induced p53-independent apoptosis. Conclusions: Our findings suggest that high-LET radiotherapy is expected to be a valid application for patients carrying mutated p53 cancer cells. We proposed that the elucidation of the p53-independent apoptosis-related genes might provide new insights into radiotherapy for cancer

  8. Effect of baicalin-copper on the induction of apoptosis in human hepatoblastoma cancer HepG2 cells.

    Science.gov (United States)

    Li, Xiaoli; Zou, Kaili; Gou, Jing; Du, Qin; Li, Dejuan; He, Xiaoyan; Li, Zhubo

    2015-03-01

    The medical properties of baicalin have been well known for many years. However, the discovery that baicalin in the presence of metal ions is more effective than baicalin alone changed the course of drug research. The present study was designed to investigate the effect and possible mechanism of apoptosis induced by baicalin-copper in a human hepatoblastoma cancer cell line (HepG2) and in vivo. This study demonstrated that baicalin-copper suppresses the proliferation of HepG2 cells in a dose-dependent manner. Intraperitoneal injection of baicalin-copper resulted in a significant decrease in tumor growth in xenografts in nude mice. Acridine orange staining and flow cytometry analysis demonstrated that baicalin-copper induced apoptosis in HepG2 cells and caused cells to arrest in G2-M phase of the cell cycle. Furthermore, baicalin-copper treatment significantly increased the Bax/Bcl-2 ratio and p38 levels, as well as decreased the expression of caspase-3, p-PI3K, p-Akt and p-mTOR (P copper induces apoptosis in HepG2 cells by down-regulating the PI3K/Akt/mTOR signaling pathway.

  9. Cytotoxicity, genotoxicity and oxidative stress of malachite green on the kidney and gill cell lines of freshwater air breathing fish Channa striata.

    Science.gov (United States)

    Majeed, S Abdul; Nambi, K S N; Taju, G; Vimal, S; Venkatesan, C; Hameed, A S Sahul

    2014-12-01

    The cytotoxicity, genotoxicity and oxidative stress of malachite green (MG) was investigated using the fish Channa striata kidney (CSK) and Channa striata gill (CSG) cell lines. Five concentrations ranging from 0.001 to 10 μg mL(-1) were tested in three independent experiments. Cytotoxicity was assessed by 3-(4, 5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay, Rhodamine 123 and Alamar Blue. The mitochondrial changes and apoptosis of MG-exposed cells were observed by Rhodamine 123 and acridine orange/ethidium bromide (AO/EB) staining, respectively. In vitro potential DNA damaging effect of MG was tested using comet assay. Mitochondrial damage, apoptosis and DNA fragmentation increased in a concentration-dependent manner. Additionally, DNA electrophoretic mobility experiments were carried out to study the binding effect of MG to double-stranded DNA (dsDNA) of cells. DNA shift mobility experiments showed that MG is capable of strongly binding to linear dsDNA causing its degradation. Biochemical parameters such as lipid peroxidation (MDA), catalase (CAT) activity and reduced glutathione (GSH) levels were evaluated after exposure to MG. In CSK and CSG cell lines exposed to MG for 48 h, a significant increase in lipid peroxidation, which might be associated with decreased levels of reduced glutathione and catalase activity in these cell lines (p < 0.001), was observed. PMID:25023653

  10. APPLICATION OF METAL RESISTANT BACTERIA BY MUTATIONAL ENHANCMENT TECHNIQUE FOR BIOREMEDIATION OF COPPER AND ZINC FROM INDUSTRIAL WASTES

    Directory of Open Access Journals (Sweden)

    M. R. Shakibaie ، A. Khosravan ، A. Frahmand ، S. Zare

    2008-10-01

    Full Text Available In this research, using mutation in the metal resistant bacteria, the bioremediation of the copper and zinc from copper factory effluents was investigated. Wastewater effluents from flocculation and rolling mill sections of a factory in the city of Kerman were collected and used for further experiments. 20 strains of Pseudomonas spp. were isolated from soil and effluents surrounding factory and identified by microbiological methods. Minimum inhibitory concentrations for copper (Cu and zinc (Zn were determined by agar dilution method. Those strains that exhibited highest minimum inhibitory concentrations values to the metals (5mM were subjected to 400-3200 mg/L concentrations of the three mutagenic agents, acriflavine, acridine orange and ethidium bromide. After determination of subinhibitory concentrations, the minimum inhibitory concentrations values for copper and zinc metal ions were again determined, which showed more than 10 fold increase in minimum inhibitory concentrations value (10 mM for Cu and 20 mM for Zn with P≤0.05. The atomic absorption spectroscopy of dried biomass obtained from resistant strains after exposure to mutagenic agents revealed that strains 13 accumulate the highest amount of intracellular copper (0.35% Cu/mg dried biomass and strain 10 showed highest accumulation of zinc (0.3% Zn/mg dried biomass respectively with P≤0.05. From above results it was concluded that the treatment of industrial waste containing heavy metals by artificially mutated bacteria may be appropriate solution for effluent disposal problems.

  11. Depletion of ribosomal protein L8 impairs Drosophila development and is associated with apoptosis

    Institute of Scientific and Technical Information of China (English)

    2010-01-01

    Ribosomal protein L8 is a component of the 60S subunit of the ribosome and is involved in protein synthesis but its role in Drosophila development is not well understood.We depleted L8 through RNA interference (RNAi) to examine its effects on fly development both in vivo and in vitro.The results demonstrated that L8 RNAi caused embryonic or first-larval lethality,delay of larval development,defects in eye and wing morphology,and dramatically reduced the number of S2 cells.This indicated that L8 plays a crucial role in Drosophila development.Acridine orange staining of the wing discs showed that apoptosis occurred when L8 was depleted,indicating that depletion of L8 is tightly connected to apoptosis.RT-PCR analyses of the transcription level of genes that are known to be key factors in apoptosis (p53,hid,reaper,dark,Dcp-1) and cell cycle regulation (cdc45,MCM3,cyclin B,incenp) in L8-deficient S2 cells,were consistent with their role in apoptosis induction and cell cycle arrest.These results indicate that depletion of L8 strongly impairs Drosophila development,and that this depletion is associated with cell proliferation arrest and apoptosis,in which p53 may play a central role.

  12. Cryopreservation of white-tailed deer (Odocoileus virginianus) semen using soybean-, liposome-, and egg yolk-based extenders.

    Science.gov (United States)

    Stewart, Jamie L; Shipley, Clifford F; Katich, Ashley Seder; Po, Eleonora; Ellerbrock, Robyn E; Lima, Fabio S; Canisso, Igor F

    2016-08-01

    The objectives of the present study were to compare the use of soybean-based (Andromed), liposome-based (Optixcell), and egg yolk-based (Ovine Red, Triladyl, and Biladyl) extenders for cryopreservation of white-tailed deer semen. In experiment 1, ejaculates obtained from six bucks were aliquoted into the following extenders: Andromed, Ovine Red, Triladyl, and Biladyl (containing 4%, 6%, or 8% of glycerol). In experiment 2, ejaculates obtained from eight bucks were divided amongst Andromed, Ovine Red, and Optixcell extenders. Total and progressive sperm motility were assessed for each sample before and after cryopreservation using a computer-automated semen analyzer. In experiment 2, flow cytometry was used for post-thaw assessment of sperm viability (SYBR-14/PI), acrosome integrity (FITC-PNA/PI), and chromatin stability (acridine orange). In experiment 1, both Andromed and Ovine Red extenders exhibited higher post-thaw total motility than Biladyl containing 4% or 6% of glycerol (pwhite-tailed semen as egg yolk-based Ovine Red extender, but are superior to egg yolk-based Biladyl or Triladyl extenders. PMID:27287191

  13. Low-cost computing and network communication for a point-of-care device to perform a 3-part leukocyte differential

    Science.gov (United States)

    Powless, Amy J.; Feekin, Lauren E.; Hutcheson, Joshua A.; Alapat, Daisy V.; Muldoon, Timothy J.

    2016-03-01

    Point-of-care approaches for 3-part leukocyte differentials (granulocyte, monocyte, and lymphocyte), traditionally performed using a hematology analyzer within a panel of tests called a complete blood count (CBC), are essential not only to reduce cost but to provide faster results in low resource areas. Recent developments in lab-on-a-chip devices have shown promise in reducing the size and reagents used, relating to a decrease in overall cost. Furthermore, smartphone diagnostic approaches have shown much promise in the area of point-of-care diagnostics, but the relatively high per-unit cost may limit their utility in some settings. We present here a method to reduce computing cost of a simple epi-fluorescence imaging system using a Raspberry Pi (single-board computer, USB color camera in conjunction with a leukocyte-selective vital dye (acridine orange) in order to determine a leukocyte count and differential from a low volume (<20 microliters) of whole blood obtained via fingerstick. Additionally, the system utilizes a "cloud-based" approach to send image data from the Raspberry Pi to a main server and return results back to the user, exporting the bulk of the computational requirements. Six images were acquired per minute with up to 200 cells per field of view. Preliminary results showed that the differential count varied significantly in monocytes with a 1 minute time difference indicating the importance of time-gating to produce an accurate/consist differential.

  14. In vitro evaluation of the cytotoxic and genotoxic effects of artemether, an antimalarial drug, in a gastric cancer cell line (PG100).

    Science.gov (United States)

    Alcântara, Diego Di Felipe Ávila; Ribeiro, Helem Ferreira; Cardoso, Plínio Cerqueira Dos Santos; Araújo, Taíssa Maíra Thomaz; Burbano, Rommel Rodriguez; Guimarães, Adriana Costa; Khayat, André Salim; de Oliveira Bahia, Marcelo

    2013-02-01

    Artemisinin is a sesquiterpene lactone endoperoxide, obtained from Artemisia annua, and extensively used as an antimalarial drug. Many studies have reported the genotoxic and cytotoxic effects of artemisinins; however, there are no studies that compare such effects between cancer cell lines and normal human cells after treatment with artemether, an artemisinin derivative. Gastric cancer is the fourth most frequent type of cancer and the second highest cause of cancer mortality worldwide. Thus, the aim of this study was to evaluate the in vitro genotoxic and cytotoxic effects induced by artemether in gastric cancer cell line (PG100) and compare them with the results obtained in human lymphocytes exposed to the same conditions. We used MTT (3-(4,5-methylthiazol-2-yl)-2, 5-diphenyl-tetrazolium bromide) assay, comet assay and ethidium bromide/acridine orange viability staining to evaluate the cytotoxic and genotoxic effects of artemether in PG100. MTT assay showed a decrease in the survival percentages for both cell types treated with different concentrations of artemether (P cancer cell line PG100. PMID:21953315

  15. Time-domain measurement of fluorescence lifetime variation with pH

    Science.gov (United States)

    Ryder, Alan G.; Power, Sarah; Glynn, Thomas J.; Morrison, John J.

    2001-07-01

    Advances in the design and miniaturization of the lasers and electronics required for Time Correlated Single Photon Counting (TCSPC) measurement of fluorescence lifetime have simplified the use of the time domain method. We have assembled a compact portable system that is capable of measuring lifetimes down to approximately 200 ps (with deconvolution) and that can operate at a range of excitation and emission wavelengths. The excitation sources are pulsed LEDs and laser diodes with a maximum pulse rate of 40 MHz and are easily interchanged. Furthermore, the development of violet and blue GaN LEDs and laser diodes is expanding the range of fluorophores available for fluorescence lifetime measurement of ion concentrations. pH sensitive fluorophores have a wide range of biological and clinical applications. The use of fluorescence lifetime rather than intensity to measure pH has a number of advantages including the reduction of effects due to the photobleaching, scattering, and intensity variations in the excitation source. Using our compact TCSPC instrumentation we have measured the dependence of fluorescence lifetimes on pH for a range of dyes in phosphate buffer over the physiologically important range of 6.0 to 8.0. Most dyes exhibit only a small variation in lifetime (pH range; however, acridine exhibits a large variation in lifetime and hence shows promise as a pH indicator.

  16. Zingiber officinale, Piper retrofractum and Combination Induced Apoptosis and p53 Expression in Myeloma and WiDr Cell Lines

    Directory of Open Access Journals (Sweden)

    HENY EKOWATI

    2012-09-01

    Full Text Available In previous studies, Zingiber officinale, Piper retrofractum, and the combination showed cytotoxic activity, induced apoptosis, and p53 expression of HeLa, T47D, and MCF-7 cell lines. This study was conducted to investigate the cytotoxic and apoptotic activity of Zingiber officinale (ZO, Piper retrofractum (PR, and the combination as well as their effect to p53 expression on Myeloma and WiDr cells. The powder of ZO, PR, and ZO + PR combination (1:1 were macerated with 96% ethanol for 3 x 24 hours. MTT cytotoxic assay was performed on Myeloma and WiDr cell lines. Apoptotic cells were stained with ethidium bromide and acridine orange. Imunohistochemical expression of p53 was examined on Myeloma and WiDr cell lines. Doxorubicin was used as positive control in all assays. Results showed that ZO, PR, and ZO + PR combination had cytotoxic activity on Myeloma cells with IC50 of 28, 36, and 55 mg/ml respectively and WiDr cell lines with IC50 of 74, 158, and 64 mg/ml respectively, induced apoptotic activity, and increased p53 expression on Myeloma and WiDr cells. These results suggest that ZO, PR, and their combination induced Myeloma and WiDr cells in apoptosis through p53 expression.

  17. Effect of Prototheca zopfii on neutrophil function from bovine milk.

    Science.gov (United States)

    Cunha, Luciane T; Pugine, Silvana P; Valle, Claudia R; Ribeiro, Andrea R; Costa, Ernane J X; De Melo, Mariza P

    2006-12-01

    This study was carried to investigate neutrophil function in the presence of Prototheca zopfii. For this purpose, bovine milk neutrophils were incubated in the absence (control) of and presence of P. zopfii, and then they were examined hydrogen peroxide (H(2)O(2)) production, antioxidant enzyme activities, and phagocytic capacity. Milk was collected from negative "California Mastitis Test" (CMT) quarter from three lactating Holstein cows after induction of leukocytosis with an intramammary infusion of oyster glycogen. H(2)O(2) production was measured using the phenol red method. Catalase activity was measured following H(2)O(2) reduction at 240 nm and the activity of glutathione reductase was determined by measuring the rate of NADPH oxidation at 340 nm. P. zopfii death was assessed by fluorescent microscopy using acridine orange assay and by colony forming units (CFUs). Comparisons between the groups were initially performed by analysis of variance (ANOVA). Significant differences were then compared using Tukey's test with a significance coefficient of 0.05. Hydrogen peroxide production, catalase and glutathione reductase activities by neutrophils incubated in presence of P. zopfii were stimulated five times, 21% and 27% respectively, compared to the unstimulated-neutrophils. Neutrophils did not affect P. zopfii death as shown by microscopy and CFUs. These observations led to the conclusion that the P. zopfii promote a high increase of H(2)O(2) production by neutrophils from bovine milk during algae exposition accompanied by increase of antioxidant enzyme activities; however, this process did not affect P. zopfii death. PMID:17146586

  18. A rapid, simple and sensitive flow cytometric system for detection of Plasmodium falciparum.

    Science.gov (United States)

    Saito-Ito, A; Akai, Y; He, S; Kimura, M; Kawabata, M

    2001-11-01

    We have established a rapid, simple and sensitive flow cytometric system for the detection of Plasmodium falciparum that involves lysing erythrocytes and staining parasites at the same time using a newly developed hemolysing and staining solution containing dodecyl methyl ammonium chloride and acridine orange. In this system, freed parasites of P. falciparum could be plotted separately from erythrocyte ghosts, white blood cells and platelets on the two-dimensional scattergram of forward-angle light scatter and green fluorescence by flow cytometry with an argon laser. It took only 2-3 min per sample to obtain the scattergram and analyze the data, including the time of sample preparation for flow cytometric analysis. Sample preparation with this method does not require any difficult handling procedures. The threshold of parasite detection was almost equal to that of microscopic examination for cultured P. falciparum. The results of drug-susceptibility assays using this system were also almost identical to those obtained using microscopic examination. In this system, parasites at different erythrocytic stages could be easily distinguished. This system must prove useful and practical for basic laboratory studies of P. falciparum including those requiring the differential measurement of parasites at specific erythrocytic stages. PMID:11719111

  19. In vitro cytotoxicity assessment of imidazolium ionic liquids: biological effects in fish Channel Catfish Ovary (CCO) cell line.

    Science.gov (United States)

    Radošević, Kristina; Cvjetko, Marina; Kopjar, Nevenka; Novak, Rudjer; Dumić, Jerka; Srček, Višnja Gaurina

    2013-06-01

    Increasing interest in the application of ionic liquids as green replacement for volatile organic solvents emphasized the need for the evaluation of their toxic effects at different biological systems in order to reduce the risk for human health and environment. To our knowledge, effects of imidazolium ionic liquids on cellular level of fish cell lines have not been studied yet. The cytotoxicity of imidazolium ionic liquids containing different anions and alkyl chain lengths as the substituent at the cation ring towards the fish CCO cell line was determined by WST-1 proliferation assay. Morphological alterations were examined by fluorescent microscopy using acridine orange/ethidium bromide staining and flow cytometry analysis was also performed. The results showed concentration-dependent cytotoxicity of ionic liquids in CCO cells, related to the type of anion and alkyl chain length, while EC50 values showed moderate to high cytotoxicity of tested imidazolium ionic liquids. Distinct morphological changes observed under fluorescence microscope and data obtained by flow cytometry suggest that the toxicity of imidazolium ionic liquids with longer alkyl chains could be related to necrosis. Results presented in here may be helpful for filling existing gaps of knowledge about ionic liquids toxicity and their impact on aquatic environment.

  20. Isolation of a flavonoid, apigenin 7-O-glucoside, from Mentha longifolia (L.) Hudson subspecies longifolia and its genotoxic potency.

    Science.gov (United States)

    Gulluce, Medine; Orhan, Furkan; Yanmis, Derya; Arasoglu, Tulin; Guvenalp, Zuhal; Demirezer, Lutfiye Omur

    2015-09-01

    Mentha is a medicinal and aromatic plant belonging to the Lamiaceae family, which is widely used in food, flavor, cosmetic and pharmaceutical industries. Recently, it has been found that the use of Mentha as a pharmaceutical source is based on its phytochemical constituents that have far been identified as tannins, saponins, phenolic acids and flavonoids. This study was designed to evaluate the mutagenic and antimutagenic activities of apigenin 7-O-glucoside (A7G), a flavonoid isolated from Mentha longifolia (L.) Hudson subspecies longifolia (ML). The possible antimutagenic potential of A7G was examined against mutagens ethyl methanesulfonate and acridine in an eukaryotic cell system Saccharomyces cerevisiae and sodium azide in Salmonella typhimurium TA1535 and 9-aminoacridine in S. typhimurium TA1537. According to our findings, any concentrations of the A7G used did not show mutagenic activity but exerted strong antimutagenic activities at tested concentrations. The inhibition rates for the Ames test ranged from 27.2% (S. typhimurium TA1535: 0.4 μM/plate) to 91.1% (S. typhimurium TA1537: 0.2 μM/plate) and for the yeast deletion assay from 4% to 57.7%. This genotoxicological study suggests that a flavonoid from ML owing to antimutagenic properties is of great pharmacological importance and might be beneficial to industries producing food additives, cosmetics and pharmaceuticals products. PMID:23377117

  1. Application of laser scanning microscopy for the analysis of oral biofilm dissolution by different endodontic irrigants

    Directory of Open Access Journals (Sweden)

    Aldo del Carpio-Perochena

    2014-01-01

    Full Text Available Background: Multi-specie biofilms are highly resistant to antimicrobials due to cellular interactions found in them. The purpose of this study was to evaluate, by confocal laser scanning microscopy, the biofilm dissolution effectiveness of different irrigant solutions on biofilms developed on infected dentin in situ. Materials and Methods: A total of 120 bovine dentin specimens infected intraorally (30/group were treated by the following solutions: 2% of chlorhexidine digluconate, 1%, 2.5% and 5.25% of sodium hypochlorite (NaOCl. The solutions were utilized for 5, 15 and 30 min with 2 experimental volumes 500 μL and 1 mL. All the samples were stained using an acridine orange and the biofilm thickness before (control group and after the experiments were evaluated, utilizing a confocal microscope at ×40. The Mann-Whitney U and the nom-parametric Kruskal-Wallis Dunns tests were utilized to determine the influence of the volume and to perform the comparisons among the groups respectively. The significance level was set at P 0.05. The biofilm dissolution treated with 1% NaOCl was directly proportional to the exposure time (P 0.05. Conclusion: The higher exposure times and concentrations of NaOCl were not sufficient to dissolve 100% of the biofilm. However, all NaOCl solutions were more effective than 2% chlorhexidine digluconate to dissolve organic matter.

  2. Cytocompatibility, cytotoxicity and genotoxicity analysis of dental implants

    International Nuclear Information System (INIS)

    Several types of materials are frequently used for dental prostheses in dental medicine. Different treatments with titanium are the most used. The aim of the present study was to analyze by means of cytotoxicity and cytocompatibility techniques the capacity of dental implants to integrate to the bone tissue. Cultures of UMR 106 cell line derived from an osteosarcoma were used for bioassays mainly because they show many of the properties of osteoblasts. Dental implant samples provided by B and W company were compared with others of recognized trademarks. The first ones contain ASTM titanium (8348 GR2) with acid printing. Cytotoxicity was analyzed by means of lysosome activity, using the neutral red technique and alkaline phosphatase enzyme activity. Cell variability was determined by means of the acridine ethidium-orange bromide technique. One-way ANOVA and Bonferroni and Duncan post-ANOVA tests were used for the statistical analysis. The assays did not show significant differences among the dental implants analyzed. Our findings show that the dental prostheses studied present high biocompatibility, quantified by the bioassays performed. The techniques employed revealed that they can be a useful tool for the analysis of other materials for dental medicine use

  3. Experimental Study of Rat Beta Islet Cells Cultured under Simulated Microgravity Conditions

    Institute of Scientific and Technical Information of China (English)

    Chun SONG; Xiu-Qing DUAN; Xi LI; Li-Ou HAN; Ping XU; Chun-Fang SONG; Lian-Hong JIN

    2004-01-01

    To observe the effects of simulated microgravity on beta islet cell culture, we have compared the survival rates and the insulin levels of the isolated rat islet cells cultured at micro- and normal gravity conditions. The survival rates of the cells cultured were determined by acridine orange-propidium iodide double-staining on day 3, 7 and 14. The morphology of the cells was observed by electron microscopy.Insulin levels were measured by radio immuno assays. Our results show that the cell number cultured under the microgravity condition is significantly higher than that under the routine condition (P<0.01). Some tubular structure shown by transmission electron microscopy, possibly for the transport of nutrients, were formed intercellularly in the microgravity cultured group on day 7. There were also abundant secretion particles and mitochondria in the cytoplasm of the cells. Scanning electron microscopy showed that there were holes formed between each islet, possibly connecting with the nutrient transport tubules. The microgravity cultured group also has higher insulin levels in the media as compared with the control group(P<0.01). Our results indicate that microgravity cultivation of islet cells has advantages over the routine culture methods.

  4. Antiproliferative and proapoptotic effects of somatostatin on activated hepatic stellate cells

    Institute of Scientific and Technical Information of China (English)

    Qin Pan; Ding-Guo Li; Han-Ming Lu; Liang-Yong Lu; Yu-Qin Wang; Qin-Fang Xu

    2004-01-01

    AIM: To assess the effects of somatostatin on proliferation and apoptosis of activated rat hepatic stellate cells (HSCs).METHODS: HSCs isolated from the livers of adult SpragueDawley rats (weighing 400-500 g) by in situ perfusion and purified by single-step density gradient centrifugation with Nycodenz, became activated after 10 days' cultivation. Then the apoptotic rate of HSCs treated with different doses of somatostatin for 72 h, was assayed by acridine orange/ethidium bromide fluorescent staining, terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling, transmission electron microscopy and flow cytometry, while the proliferation of HSCs was measured by MTT assay. Furthermore, the mechanisms of somatostatin were investigated by cytodynamic analysis.RESULTS: Somatostatin at the concentration of 10-6-10-9 mol/L could decrease the proliferative rate, and promote the apoptosis of activated rat HSCs in a dose-dependent way.Its action was most significant when the concentration reached 10-6 mol/L or 10-7 mol/L (P<0.05-0.01). An obvious cell-cycle arrest (G0/G1 arrest) was the important way for somatostatin to exert its action.CONCLUSION: Antiproliferative and proapoptotic effects of low-dose somatostatin on activated rat HSCs can be obtained.These findings reveal its potential antifibrotic action.

  5. 2,3,7,8-tetrachlorodibenzo-p-dioxin induced autophagy in a bovine kidney cell line.

    Science.gov (United States)

    Fiorito, Filomena; Ciarcia, Roberto; Granato, Giovanna Elvira; Marfe, Gabriella; Iovane, Valentina; Florio, Salvatore; De Martino, Luisa; Pagnini, Ugo

    2011-12-18

    The administration of 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD) to a variety of cultured cells may alter their ability to proliferate and die. In a previous study we demonstrated that TCDD induced proliferation in Madin-Darby Bovine Kidney (MDBK) cells where no signs of apoptosis were observed, but herein, analysis of MDBK cell morphology, in a large number of exposed cells, revealed some alterations, as expanded cytoplasm, an increase of intercellular spaces and many pyknotic nuclei. Hence, the aim of the current study was to elucidate the influences of dioxin on cell proliferation and cell death. We found that dioxin increased proliferation, as well as, activated cell death with autophagy, as we detected by increased amount of LC3-II, an autophagosome marker. Furthermore, formation of acidic vesicular organelles was observed by fluorescence microscopy following staining with the lysosomotropic agent acridine orange. These results were accompanied by down-regulation of telomerase activity, bTERT and c-Myc. Key tumor-suppressor protein p53 and expression of cell cycle inhibitor p21Waf1/Cip1 were activated after TCDD exposure. These changes occurred with activation of ATM phosphorylation in the presence of a decrease in Mdm2 protein levels. Taken together, these results support the idea that TCDD in MDBK cells, may exert its action, in part, by enhancing cell proliferation, but also by modulating the incidence of induced cell death with autophagy.

  6. Reliable Screening of Dye Phototoxicity by Using a Caenorhabditis elegans Fast Bioassay.

    Science.gov (United States)

    Bianchi, Javier Ignacio; Stockert, Juan Carlos; Buzzi, Lucila Ines; Buzz, Lucila Ines; Blázquez-Castro, Alfonso; Simonetta, Sergio Hernán

    2015-01-01

    Phototoxicity consists in the capability of certain innocuous molecules to become toxic when subjected to suitable illumination. In order to discover new photoactive drugs or characterize phototoxic pollutants, it would be advantageous to use simple biological tests of phototoxicy. In this work, we present a pilot screening of 37 dyes to test for phototoxic effects in the roundworm Caenorhabditis elegans. Populations of this nematode were treated with different dyes, and subsequently exposed to 30 min of white light. Behavioral outcomes were quantified by recording the global motility using an infrared tracking device (WMicrotracker). Of the tested compounds, 17 dyes were classified as photoactive, being phloxine B, primuline, eosin Y, acridine orange and rose Bengal the most phototoxic. To assess photoactivity after uptake, compounds were retested after washing them out of the medium before light irradiation. Dye uptake into the worms was also analyzed by staining or fluorescence. All the positive drugs were incorporated by animals and produced phototoxic effects after washing. We also tested the stress response being triggered by the treatments through reporter strains. Endoplasmic reticulum stress response (hsp-4::GFP strain) was activated by 22% of phototoxic dyes, and mitochondrial stress response (hsp-6::GFP strain) was induced by 16% of phototoxic dyes. These results point to a phototoxic perturbation of the protein functionality and an oxidative stress similar to that reported in cell cultures. Our work shows for the first time the feasibility of C. elegans for running phototoxic screenings and underscores its application on photoactive drugs and environmental pollutants assessment.

  7. The photoreduction and photosensitized reduction of dyes bound to a surfactant micellar surface

    Energy Technology Data Exchange (ETDEWEB)

    Usui, Y.; Saga, K.

    1982-10-01

    The photochemical reduction of thionine(Th) and Methylene Blue(MB) bound to anionic sodium dodecyl sulfate(SDS) micelles in an aqueous solution was inhibited at the lower concentration range of a reductant anion, EDTA, because of the electrostatic repulsion between the micelles and the reductant. The concentration dependence of EDTA on the quantum yield of photoreduction has shown to be, anomalously a sigmoidal type: this is explained by mechanistic interpretation based on the cooperative effects of the ionic strength and the essential concentration of EDTA. The enhancement effect for the quantum yield of the photoreduction of eosine bound to a cationic micelle by the EDTA anion was conversely observed, because of the electrostatic attraction. When 10-dodecyl) (Acridine Orange) was incorporated into SDS micelles in water containing EDTA and ascorbic acid, and the mixture was excited, the photosensitized reduction reduction of MB bound to the micelles occurred. It was found that the quantum yield of the sensitized reduction depended on the concentration of MB and was larger than the yield on the direct excitation of MB.

  8. Trichomoniasis: An update.

    Science.gov (United States)

    Preethi, V; Mandal, Jharna; Halder, Ajay; Parija, Subhash Chandra

    2011-07-01

    Trichomoniasis is one of the most common sexually transmitted infections (STIs) with a prevalence of 5-75%. In India, trichomoniasis accounts for 2-7% of all STIs. Infection with Trichomonas vaginalis is known to cause vaginitis. Significant association has also been noted between trichomoniasis and cervical cancer, atypical pelvic inflammatory disease, infertility, low birth weight, and respiratory-tract infection in neonates. Of interest are the recent documentations of association of this parasite with human immunodeficiency virus. Use of fluorescent dyes such as acridine orange has increased the sensitivity of the direct microscopy. Culture has been found to be more sensitive than the direct microscopy but has its own limitations. Antigen detection systems have hastened the proce ss of diagnosis tremendously. Molecular methods have been found to be very sensitive and specific. Once the presence of T. vaginalis has been documented, other STIs should also be actively looked for in that particular individual. Therapy should involve both the partners for proper control and eradication of the disease. PMID:23508486

  9. Cytotoxic Effects of Newly Synthesized Palladium(II Complexes of Diethyldithiocarbamate on Gastrointestinal Cancer Cell Lines

    Directory of Open Access Journals (Sweden)

    Shahram Hadizadeh

    2014-01-01

    Full Text Available As a part of a drug development program to discover novel therapeutic and more effective palladium (Pd based anticancer drugs, a series of water-soluble Pd complexes have been synthesized by interaction between [Pd (phen(H2O2(NO32] and alkylenebisdithiocarbamate(al-bis-dtc disodium salts. This study was undertaken to examine the possible cytotoxic effect of three novel complexes (0.125–64 µg/mL on human gastric carcinoma (AGS, esophageal squamous cell carcinoma (Kyse-30, and hepatocellular carcinoma (HepG2 cell lines. The cytotoxicity was examined using cell proliferation and acridine orange/ethidium bromide (AO/EB assay. In order to examine the effects of new Pd(II complexes on cell cycle status, we performed cell cycle analysis. The complexes were found to have completely lethal effects on the cell lines, and the half maximal inhibitory concentration (IC50 values obtained for the cell lines were much lower in comparison with cisplatin. We demonstrated that the three new Pd(II complexes are able to induce G2/M phase arrest in AGS and HepG2; in addition, the Pd(II complexes caused an S phase arrest in Kyse-30 cell line. Our results indicate that newly synthesized Pd(II complexes may provide a novel class of chemopreventive compounds for anticancer therapy.

  10. Thermodynamic characterization of proflavine–DNA binding through microcalorimetric studies

    International Nuclear Information System (INIS)

    Highlights: • Energetics of the interaction of proflavine with DNA has been studied. • The binding reaction was favored by both negative enthalpy and positive entropy. • Enthalpy–entropy compensation phenomenon was observed. • Non-polyelectrolytic forces played a dominant role in the binding process. • Proflavine enhanced the thermal stability of DNA remarkably. - Abstract: The interaction of an important acridine dye, proflavine hydrochloride, with double stranded DNA was investigated using isothermal titration calorimetry and differential scanning calorimetry. The equilibrium constant for the binding reaction was calculated to be (1.60 ± 0.04) · 105 · M−1 at T = 298.15 K. The binding of proflavine hydrochloride to DNA was favored by both negative enthalpy and positive entropy contributions to the Gibbs energy. The equilibrium constant for the binding reaction decreased with increasing temperature. The standard molar enthalpy change became increasingly negative while the standard molar entropy change became less positive with rise in temperature. However, the standard molar Gibbs free energy change varied marginally suggesting the occurrence of enthalpy–entropy compensation phenomenon. The binding reaction was dominated by non-polyelectrolytic forces which remained virtually unchanged at all the salt concentrations studied. The binding also significantly increased the thermal stability of DNA against thermal denaturation

  11. Anticancer Activity of Indian Stingless Bee Propolis: An In Vitro Study

    Directory of Open Access Journals (Sweden)

    Milind K. Choudhari

    2013-01-01

    Full Text Available Indian stingless bee propolis has a complex chemical nature and is reported to possess various medicinal properties. In the present study, anticancer activity of the ethanolic extract of propolis (EEP was explored by testing the cytotoxic and apoptotic effect in four different cancer cell lines, namely, MCF-7 (human breast cancer, HT-29 (human colon adenocarcinoma, Caco-2 (human epithelial colorectal adenocarcinoma, and B16F1 (murine melanoma, at different concentrations. Cytotoxicity was evaluated by MTT assay and Trypan blue dye exclusion assay. EEP at a concentration of 250 g/mL exhibited ≥50% mortality in all cell lines tested (i.e., IC50 value. EEP revealed a concentration and time dependent cytotoxic effect. Apoptosis was estimated by differential staining (ethidium bromide/acridine orange and TUNEL (deoxynucleotidyl transferase-dUTP nick end labeling assay. Light microscopy and atomic force microscopy demonstrated morphological features of apoptosis in all the cell lines after treatment with 250 g/mL EEP for 24 h. Thus, early onset of apoptosis is the reason for anticancer activity of Indian stingless bee propolis. Further, the antioxidant potential of Indian stingless bee propolis was demonstrated to substantiate its anticancer activity.

  12. Rosiglitazone enhances fluorouracil-induced apoptosis of HT-29 cells by activating peroxisome proliferator-activated receptor γ

    Institute of Scientific and Technical Information of China (English)

    Yan-Qin Zhang; Xiao-Qing Tang; Li Sun; Lin Dong; Yong Qin; Hua-Qing Liu; Hong Xia; Jian-Guo Cao

    2007-01-01

    AIM: To examine whether and how rosiglitazone enhances apoptosis induced by fluorouracil in human colon cancer (HT-29) cells.METHODS: Human colon cancer HT-29 cells were cultured in vitro and treated with fluorouracil and/or rosiglitazone. Proliferation and growth of HT-29 cells were evaluated by MTT assay and trypan blue exclusion methods, respectively. The apoptosis of HT-29 cells was determined by acridine orange/ethidium bromide staining and flow cytometry using PI fluorescence staining. The expressions of peroxisome proliferator-activated receptor y (PPARy), Bcl-2 and Bax in HT-29 cells were analyzed by Western blot.RESULTS: Although rosiglitazone at the concentration below 30 umol/L for 72 h exerted almost no inhibitory effect on proliferation and growth of HT-29 cells, it could significantly enhance fluorouracil-induced HT-29 cell proliferation and growth inhibition. Furthermore, 10 umol/L rosilitazone did not induce apoptosis of HT-29 cells but dramatically enhanced fluorouracil-induced apoptosis of HT-29 cells. However, rosiglitazone did not improve apoptosis induced by fluorouracil in HT-29 cells pretreated with GW9662, a PPARy antagonist. Meanwhile, the expression of Bax and PPARy was up-regulated, while the expression of Bcl-2 was down regulated in HT-29 cells treated with rosiglitazone in a time-dependent manner. However, the effect of rosiglitazone on Bcl-2 and Bax was blocked or diminished in the presence of GW9662.CONCLUSION: Rosiglitazone enhances fluorouracil-induced apoptosis of HT-29 cells by activating PPARγ.

  13. In-vitro human spermatozoa nuclear decondensation assessed by flow cytometry.

    Science.gov (United States)

    Samocha-Bone, D; Lewin, L M; Weissenberg, R; Madgar, Y; Soffer, Y; Shochat, L; Golan, R

    1998-02-01

    The process of sperm chromatin decondensation occurs when a spermatozoon enters an ovum. Protamine disulphide bonds are reduced to SH and the polycationic protamines combine with the polyanionic egg protein, nucleoplasmin, thus being stripped from DNA which then combines with histones. Defective chromatin decondensation will thus prevent further development of the male pronucleus. In this study human sperm samples were incubated in vitro at 28 degrees C (using a medium in which the polyanion, heparin, substitutes for nucleoplasmin and beta-mercaptoethanol for egg glutathione) for 10, 20 and 30 min before stopping the reaction with formalin (to 3.6%). The DNA of the fixed cells was stained with Acridine Orange by a one-step method and subjected to flow cytometry and data analysis, in which a zone characteristic of condensed chromatin is outlined on red-green fluorescence contour plots. After 20 min of incubation 97% of the control spermatozoa that were in the mature window (WIN M) had decondensed and moved out of this region. Defects in sperm decondensation were seen in four semen samples of the 20 that were tested. In cases where spermatozoa fail to produce a fertilized egg the cause may lie with defective chromatin quality, including failure of the sperm chromatin to decondense. The method described here is a simple procedure for detecting sperm samples containing such defective cells.

  14. Catalytic degradation of recalcitrant pollutants by Fenton-like process using polyacrylonitrile-supported iron (II) phthalocyanine nanofibers: Intermediates and pathway.

    Science.gov (United States)

    Zhu, Zhexin; Chen, Yi; Gu, Yan; Wu, Fei; Lu, Wangyang; Xu, Tiefeng; Chen, Wenxing

    2016-04-15

    Iron (II) phthalocyanine (FePc) molecules were isolated in polyacrylonitrile (PAN) nanofibers by electrospinning to prevent the formation of dimers and oligomers. Carbamazepine (CBZ) and Rhodamine B (RhB) degradation was investigated during a Fenton-like process with FePc/PAN nanofibers. Classical quenching tests with isopropanol and electron paramagnetic resonance tests with 5,5-dimethyl-pyrroline-oxide as spin-trapping agent were performed to determine the formation of active species during hydrogen peroxide (H2O2) decomposition by FePc/PAN nanofibers. After eight recycles for CBZ degradation over the FePc/PAN nanofibers/H2O2 system, the removal ratios of CBZ remained at 99%. Seven by-products of RhB and twelve intermediates of CBZ were identified using ultra-performance liquid chromatography and high-resolution mass spectrometry. Pathways of CBZ and RhB degradation were proposed based on the identified intermediates. As the reaction proceeded, all CBZ and RhB aromatic nucleus intermediates decreased and were transformed to small acids, but also to potentially toxic epoxide-containing intermediates and acridine, because of the powerful oxidation ability of •OH in the catalytic system. PMID:26949842

  15. Red blood cells of the lizards Ameiva ameiva (Squamata, Teiidae) display multiple mechanisms to control cytosolic calcium.

    Science.gov (United States)

    Beraldo, F H; Sartorello, R; Gazarini, M L; Caldeira, W; Garcia, C R S

    2002-02-01

    We have previously reported that lizard red blood cells control their cytosolic calcium concentration by sequestering calcium ions in pools, which could be discharged by thapsigargin, by the Na+/H+ ionophore, monensin, by the K+/H+ ionophore, nigericin and by the proton pump inhibitor, bafilomycin A1 [1]. We have now demonstrated, with the aid of confocal microscopy, the presence in these cells of organelles, which accumulate the dye acridine orange and are thus by inference the sites of proton pools. We have found, moreover, that monensin, nigericin and bafilomycin all act to discharge these pools. We further show that calcium release ensues when the calcium ionophore, ionomycin, is added after thapsigargin and monensin; this implies the existence of a third pool, besides the acidic pool and the Endoplasmic Reticulum (ER), which participates in calcium homeostasis. The ER calcium pool can de discharged by the addition of the second messenger, IP3, and we present evidence, based on confocal microscopy, that the IP3 receptors are located in or close to the nucleus. PMID:11969248

  16. Use of immunomagnetic separation for the detection of Desulfovibrio vulgaris from environmental samples

    Energy Technology Data Exchange (ETDEWEB)

    Chakraborty, R.; Hazen, T.C.; Joyner, D.C.; Kusel, K.; Singer, M.E.; Sitte, J.; Torok, T.

    2011-04-15

    Immunomagnetic separation (IMS) has proved highly efficient for recovering microorganisms from heterogeneous samples. Current investigation targeted the separation of viable cells of the sulfate-reducing bacterium, Desulfovibrio vulgaris. Streptavidin-coupled paramagnetic beads and biotin labeled antibodies raised against surface antigens of this microorganism were used to capture D. vulgaris cells in both bioreactor grown laboratory samples and from extremely low-biomass environmental soil and subsurface drilling samples. Initial studies on detection, recovery efficiency and viability for IMS were performed with laboratory grown D. vulgaris cells using various cell densities. Efficiency of cell isolation and recovery (i.e., release of the microbial cells from the beads following separation) was followed by microscopic imaging and acridine orange direct counts (AODC). Excellent recovery efficiency encouraged the use of IMS to capture Desulfovibrio spp. cells from low-biomass environmental samples. The environmental samples were obtained from a radionuclide-contaminated site in Germany and the chromium (VI)-contaminated Hanford site, an ongoing bioremediation project of the U.S. Department of Energy. Field deployable IMS technology may greatly facilitate environmental sampling and bioremediation process monitoring and enable transcriptomics and proteomics/metabolomics-based studies directly on cells collected from the field.

  17. Luteolin decreases the attachment, invasion and cytotoxicity of UPEC in bladder epithelial cells and inhibits UPEC biofilm formation.

    Science.gov (United States)

    Shen, Xiao-fei; Ren, Lai-bin; Teng, Yan; Zheng, Shuang; Yang, Xiao-long; Guo, Xiao-juan; Wang, Xin-yuan; Sha, Kai-hui; Li, Na; Xu, Guang-ya; Tian, Han-wen; Wang, Xiao-ying; Liu, Xiao-kang; Li, Jingyu; Huang, Ning

    2014-10-01

    Urinary tract infection (UTI), primarily caused by uropathogenic Escherichia coli (UPEC), is one of the most common infectious diseases worldwide. Emerging antibiotic resistance requires novel treatment strategies. Luteolin, a dietary polyphenolic flavonoid, has been confirmed as a potential antimicrobial agent. Here, we evaluated the sub-MICs of luteolin for potential properties to modulate the UPEC infection. We found that luteolin significantly decreased the attachment and invasion of UPEC J96 or CFT073 in human bladder epithelial cell lines T24. Meanwhile, obvious decreased expression of type 1 fimbriae adhesin fimH gene, lower bacterial surface hydrophobicity and swimming motility, were observed in luteolin-pretreated UPEC. Furthermore, luteolin could attenuate UPEC-induced cytotoxicity in T24 cells, which manifested as decreased activity of lactate dehydrogenase (LDH). Simultaneously, the inhibition of luteolin on UPEC-induced cytotoxicity was confirmed by ethidium bromide/acridine orange staining. Finally, the luteolin-pretreated UPEC showed a lower ability of biofilm formation. Collectively, these results indicated that luteolin decreased the attachment and invasion of UPEC in bladder epithelial cells, attenuated UPEC-induced cytotoxicity and biofilm formation via down-regulating the expression of adhesin fimH gene, reducing the bacterial surface hydrophobicity and motility.

  18. Subcellular and in-vivo Nano-Endoscopy

    Science.gov (United States)

    Cheemalapati, Surya Venkatasekhar; Winskas, John; Wang, Hao; Konnaiyan, Karthik; Zhdanov, Arseny; Roth, Alison; Adapa, Swamy Rakesh; Deonarine, Andrew; Noble, Mark; Das, Tuhin; Gatenby, Robert; Westerheide, Sandy D.; Jiang, Rays H. Y.; Pyayt, Anna

    2016-01-01

    Analysis of individual cells at the subcellular level is important for understanding diseases and accelerating drug discovery. Nanoscale endoscopes allow minimally invasive probing of individual cell interiors. Several such instruments have been presented previously, but they are either too complex to fabricate or require sophisticated external detectors because of low signal collection efficiency. Here we present a nanoendoscope that can locally excite fluorescence in labelled cell organelles and collect the emitted signal for spectral analysis. Finite Difference Time Domain (FDTD) simulations have shown that with an optimized nanoendoscope taper profile, the light emission and collection was localized within ~100 nm. This allows signal detection to be used for nano-photonic sensing of the proximity of fluorophores. Upon insertion into the individual organelles of living cells, the nanoendoscope was fabricated and resultant fluorescent signals collected. This included the signal collection from the nucleus of Acridine orange labelled human fibroblast cells, the nucleus of Hoechst stained live liver cells and the mitochondria of MitoTracker Red labelled MDA-MB-231 cells. The endoscope was also inserted into a live organism, the yellow fluorescent protein producing nematode Caenorhabditis elegans, and a fluorescent signal was collected. To our knowledge this is the first demonstration of in vivo, local fluorescence signal collection on the sub-organelle level. PMID:27694854

  19. Low-temperature spectra of the analogues of 10-hydroxybenzo[h]quinoline as an indication of barrierless ESIPT.

    Science.gov (United States)

    Deperasińska, Irena; Gryko, Daniel T; Karpiuk, Elena; Kozankiewicz, Bolesław; Makarewicz, Artur; Piechowska, Joanna

    2012-12-13

    The absorption and fluorescence spectra of two analogues of 10-hydroxybenzo[h]quinoline (10-HBQ), namely, 1-hydroxy-7-methylbenzo[c]acridine (HMBA) and 4-hydroxybenzo[c]phenanthridine (HBPA), were studied in n-alkane matrices at 5 K. Considerable energy separation between the onsets of the spectra and broadening of the bands was an indication that intramolecular proton transfer (ESIPT) takes place at such a low temperature. DFT and ab initio methods were used to calculate the electronic transition energies and oscillator strengths and the vibronic structure of the electronic spectra. Shortcomings in our knowledge of the shape of the potential energy surface for ESIPT systems are highlighted in the context of the discussion of the shape of the electronic spectra. The π-expansion of the 10-HBQ chromophore achieved by adding a benzene moiety at various positions adjacent to the pyridine ring led to compounds possessing diverse photophysical properties, ranging from the non-ESIPT strongly fluorescent molecule of 10-hydroxy-1-azaperylene to weakly emitting (or nonemitting) molecules, where ESIPT occurs very efficiently.

  20. Apoptosis of human tongue squamous cell carcinoma cell (CAL-27 induced by Lactobacillus sp. A-2 metabolites

    Directory of Open Access Journals (Sweden)

    Guoliang ZHANG

    2014-07-01

    Full Text Available Objective: To study the effect of Lactobacillus sp. A-2 metabolites on viability of CAL-27 cells and apoptosis in CAL-27 cells. Methods: Lactobacillus sp. A-2 metabolites 1 and 2 (LM1 and LM2 were obtained by culturing Lactobacillus sp. A-2 in reconstituted whey medium and whey-inulin medium; the cultured CAL-27 cells were treated with different concentrations of LM1 and LM2 (0, 3, 6, 12, 24, 48 mg/mL and assayed by methyl thiazolyltetrazolium (MTT method; morphological changes of apoptotic cell were observed under fluorescence microscopy by acridine orange (Ao fluorescent staining; flow cytometry method (FCM and agarose gel electrophoresis were used to detect the apoptosis of CAL-27 cells treated LM1 and LM2. Results: The different concentrations of LM1 and LM2 could restrain the growth of CAL-27 cells, and in a dose-dependent manner; the apoptosis of CAL-27 cells was obviously induced and was time-dependent. Conclusions: Viability of CAL-27 cells was inhibited by Lactobacillus sp. A-2 metabolites; Lactobacillus sp. A-2 metabolites could induce CAL-27 cells apoptosis; study on the bioactive compounds in the Lactobacillus sp. A-2 metabolites and their molecular mechanism is in progress.

  1. Sperm quality improvement after natural anti-oxidant treatment of asthenoteratospermic men with leukocytospermia

    Institute of Scientific and Technical Information of China (English)

    Paola Piomboni; Laura Gambera; Francesca Serafini; Giovanna Campanella; Giuseppe Morgante; Vincenzo De Leo

    2008-01-01

    Aim: To study the immune-modulating and anti-oxidant effects of beta-glucan, papaya, lactoferrin, and vitamins C and E on sperm characteristics of patients with asthenoteratozoospermia associated with leucocytosis. Methods:Fifty-one patients referred to our Sterility Center for semen analysis were selected. Sperm parameters were assessed before and after patient's treatment with beta-glucan, lactoferrin, papaya, and vitamins C and E. DNA damage was assessed by the acridine orange test and sperm structural characteristics were evaluated by transmission electron microscopy. Results: After 90 days of treatment, an increase in the percentage of morphologically normal sperm (17.0±5.2 vs. 29.8±6.5) and total progressive motility (19.0±7.8 vs. 34.8±6.8) were detected. Structural sperm characteristics as well as chromatin integrity were also improved after treatment. In terms of leukocyte concentration in seminal fluid, a significant reduction was recorded (2.2±0.9 vs. 0.9±0.2). Conclusion: The treatment of an inflammatory process by the synergic action of immune modulators and anti-oxidants could protect sperm during maturation and migration, leading to improved sperm function.

  2. Positive selection of mutants with cell envelope defects of a Salmonella typhimurium strain hypersensitive to the products of genes hisF and hisH

    International Nuclear Information System (INIS)

    Strain SB564 and its derivative DA78 are hypersensitive to the inhibitory action of the proteins coded for by genes hisF and hisH on cell division. Transduction of hisO1242, a regulatory mutation that elicits a very high level of expression of the histidine operon, into these strains resulted in the production of long filamentous cells carrying large balloons and in growth failure. Forty-one hisO1242 derivatives that escaped inhibition were isolated. These strains showed a large variety of alterations, many of which were related to the cell envelope. The more-frequent alterations included: changes in cell shape, increased sensitivity to one or more of several drugs (deoxycholate, cycloserine, penicillin, novobiocin, acridine orange), increased autolytic activity in alkaline buffer, anomalous fermentation of maltose on eosin--methylene blue plates, and temperature-conditional cell division. The alterations are produced, in some of the strains, by pleiotropic mutations in gene envB. Strains affected in divC, divD, and rodA loci have also been identified. Genetic analaysis has shown that several strains carry more than one envelope mutation. It is assumed that envelope mutations are positively selected because they somehow alleviate the particularly severe inhibition of cell division caused, in strains SB564 and DA78, by the excessive synthesis of hisF and hisH gene products

  3. The Acetone Extract of Sclerocarya birrea (Anacardiaceae Possesses Antiproliferative and Apoptotic Potential against Human Breast Cancer Cell Lines (MCF-7

    Directory of Open Access Journals (Sweden)

    Nicoline Fri Tanih

    2013-01-01

    Full Text Available Interesting antimicrobial data from the stem bark of Sclerocarya birrea, which support its use in traditional medicine for the treatment of many diseases, have been delineated. The current study was aimed to further study some pharmacological and toxicological properties of the plant to scientifically justify its use. Anticancer activity of water and acetone extracts of S. birrea was evaluated on three different cell lines, HT-29, HeLa, and MCF-7 using the cell titre blue viability assay in 96-well plates. Apoptosis was evaluated using the acridine orange and propidium iodide staining method, while morphological structure of treated cells was examined using SEM. The acetone extract exhibited remarkable antiproliferative activities on MCF-7 cell lines at dose- and time-dependent manners (24 h and 48 h of incubation. The extract also exerted apoptotic programmed cell death in MCF-7 cells with significant effect on the DNA. Morphological examination also displayed apoptotic characteristics in the treated cells, including clumping, condensation, and culminating to budding of the cells to produce membrane-bound fragmentation, as well as formation of apoptotic bodies. The acetone extract of S. birrea possesses antiproliferative and apoptotic potential against MCF-7-treated cells and could be further exploited as a potential lead in anticancer therapy.

  4. DNA methylation detection based on difference of base content

    Science.gov (United States)

    Sato, Shinobu; Ohtsuka, Keiichi; Honda, Satoshi; Sato, Yusuke; Takenaka, Shigeori

    2016-04-01

    Methylation frequently occurs in cytosines of CpG sites to regulate gene expression. The identification of aberrant methylation of certain genes is important for cancer marker analysis. The aim of this study was to determine the methylation frequency in DNA samples of unknown length and/or concentration. Unmethylated cytosine is known to be converted to thymine following bisulfite treatment and subsequent PCR. For this reason, the AT content in DNA increases with an increasing number of methylation sites. In this study, the fluorescein-carrying bis-acridinyl peptide (FKA) molecule was used for the detection of methylation frequency. FKA contains fluorescein and two acridine moieties, which together allow for the determination of the AT content of double-stranded DNA fragments. Methylated and unmethylated human genomes were subjected to bisulfide treatment and subsequent PCR using primers specific for the CFTR, CDH4, DBC1, and NPY genes. The AT content in the resulting PCR products was estimated by FKA, and AT content estimations were found to be in good agreement with those determined by DNA sequencing. This newly developed method may be useful for determining methylation frequencies of many PCR products by measuring the fluorescence in samples excited at two different wavelengths.

  5. Implementation of fluorescence confocal mosaicing microscopy by "early adopter" Mohs surgeons: a review of recent progress

    Science.gov (United States)

    Jain, Manu; Rajadhyaksha, Milind; Nehal, Kishwer

    2016-03-01

    Confocal mosaicing microscopy (CMM) enables rapid imaging of large areas of fresh tissue ex vivo without the processing that is necessary for conventional histology. When performed with fluorescence mode using acridine orange (nuclear specific dye) it enhances nuclei-to-dermis contrast that enables detection of all types of BCCs including thin strands of infiltrative basal cell carcinomas (BCCs). Thus far, this technique has been mostly validated in research setting for the analysis of BCC tumor margins. Recently, CMM has been adopted and implemented in real clinical settings by some surgeons as an alternative tool to frozen section (FS) during Mohs surgery. In this review article we summarize the development of CMM guided imaging of ex vivo tissues from bench to bedside. We also present its current state of application in routine clinical workflow not only for the assessment of BCC margin but also for other skin cancers such as melanoma, SCC, and some infectious diseases where FS is not routinely performed. Lastly, we also discuss the potential limitations of this technology as well as future developments. As this technology advances further, it may serve as an adjunct to standard histology and enable rapid surgical pathology of skin cancers at the bedside.

  6. Inhibitory Effects of Mistletoe Alkali on Salivary Adenoid Cystic Carcinoma Cells

    Institute of Scientific and Technical Information of China (English)

    LI Mei-hua; WANG Yi-shu; ZHOU Hong-lan; LI Ya-juan; QIU Xin-ru; WANG Xue-yao; ZHAO Yu-yang

    2013-01-01

    Mistletoe alkali plays an important role in salivary adenoid cystic carcinoma(SACC) cell proliferation,apoptosis and invasion.Mistletoe alkali shows potent anticaner property.In this paper,immunocytochemical and immunofluorescence staining were employed to evaluate the expression levels of proliferating cell nuclear antigen(PCNA),Caspase 3,Caspase 8 and Caspase 9.Apoptosis was detected by acridine orange/ethidium bromide (AO/EB) staining,cell invasion ability was assessed by Boyden Chamber assay.Pretreatment with mistletoe alkali markedly decreased PCNA expression in SACC cells in a dose-dependent manner(P<0.001) and also led to increase the expression of Caspase 3,Caspase 8 and Caspase 9 in SACC cells compared with control group(P<0.001).Number of apoptotic cells increased dramatically in mistletoe alkali group(P<0.001).In Boyden Chamber assay,mistletoe alkali treatment could inhibit SACC cells to penetrate the artificial basement membrane compared with control group(P<0.01).Mistletoe alkali remarkably inhibited the proliferation and invasion of SACC cells and induced the apoptosis of SACC cells.These results provide an insight into the mechanisms of anticancer effects of mistletoe alkali,and highlight the potential clinical application of it.

  7. Interaction of coumarin with calf thymus DNA: deciphering the mode of binding by in vitro studies.

    Science.gov (United States)

    Sarwar, Tarique; Rehman, Sayeed Ur; Husain, Mohammed Amir; Ishqi, Hassan Mubarak; Tabish, Mohammad

    2015-02-01

    DNA is the major target for a wide range of therapeutic substances. Thus, there has been considerable interest in the binding studies of small molecules with DNA. Interaction between small molecules and DNA provides a structural guideline in rational drug designing and in the synthesis of new and improved drugs with enhanced selective activity and greater clinical efficacy. Plant derived polyphenolic compounds have a large number of biological and pharmacological properties. Coumarin is a polyphenolic compound which has been extensively studied for its diverse pharmacological properties. However, its mode of interaction with DNA has not been elucidated. In the present study, we have attempted to ascertain the mode of binding of coumarin with calf thymus DNA (Ct-DNA) through various biophysical techniques. Analysis of UV-visible absorbance spectra and fluorescence spectra indicates the formation of complex between coumarin and Ct-DNA. Several other experiments such as effect of ionic strength, iodide induced quenching, competitive binding assay with ethidium bromide, acridine orange and Hoechst 33258 reflected that coumarin possibly binds to the minor groove of the Ct-DNA. These observations were further supported by CD spectral analysis, viscosity measurements, DNA melting studies and in silico molecular docking.

  8. Cardenolide-Induced Lysosomal Membrane Permeabilization Demonstrates Therapeutic Benefits in Experimental Human Non-Small Cell Lung Cancers

    Directory of Open Access Journals (Sweden)

    Tatjana Mijatovic

    2006-05-01

    Full Text Available Non-small cell lung cancers (NSCLCs are the leading cause of cancer deaths in most developed countries. Targeting heat shock protein 70 (Hsp70 expression and function, together with the induction of lysosomal membrane permeabilization (LMP, could overcome the multiple anti-cell death mechanisms evidenced in NSCLCs that are responsible for the failure of currently used chemotherapeutic drugs. Because cardenolides bind to the sodium pump, they affect multiple signaling pathways and thus have a number of marked effects on tumor cell behavior. The aim of the present study was to characterize in vitro and in vivo the antitumor effects of a new cardenolide (UNBS1450 on experimental human NSCLCs. UNBS1450 is a potent source of in vivo antitumor activity in the case of paclitaxeland oxaliplatin-resistant subcutaneous human NCIH727 and orthotopic A549 xenografts in nude mice. In vitro UNBS1450-mediated antitumor activity results from the induction of nonapoptotic cell death. UNBS1450 mediates the decrease of Hsp70 at both mRNA and protein levels, and this is at least partly due to UNBS1450-induced downregulation of NFAT5/ TonEBP (a factor responsible for the transcriptional control of Hsp70. These effects were paralleled by the induction of LMP, as evidenced by acridine orange staining and immunofluorescence analysis for cathepsin B accumulation.

  9. Effects of tamoxifen citrate on gene expression during nuclear chromatin condensation in male rats

    Institute of Scientific and Technical Information of China (English)

    Mukhtar Aleem; Varsha Padwal; Jyoti Choudhari; Nafisa Balasinor; Priyanka Parte; Manjeet Gill-Sharma

    2005-01-01

    Aim: To evaluate the effects of tamoxifen citrate on gene expression during nuclear chromatin condensation in male decondensation, acridine orange (AO) dye uptake, concentration of thiol-groups, levels and/or expression of transition proteins 1, 2 (TP1, TP2), protamine 1 (P1), cyclic AMP response element modulator-τ (CREMτ), androgenbinding protein (ABP) and cyclic adenosine 3', 5' monophosphate (cAMP) were evaluated after 60 days of exposure in adult male rats. Controls received the vehicle. Results: Tamoxifen citrate enhanced the rates of chromatin decondensation, increased AO dye uptake and reduced free thiols in caput epididymal sperms and reduced the levels of TP1, TP2, P1, and CREMτ in the testis, while cAMP was unaffected. P1 deposition was absent in the sperm. The transcripts of TP1, TP2 were increased, of P1 and ABP decreased, while those of CREMτ unaffected in the testis.Conclusion: Tamoxifen citrate reduced caput epididymal sperm chromatin compaction by reducing the testicular levels of proteins TP1, TP2 and P1 and the CREMτ involved in chromatin condensation during spermiogenesis.Tamoxifen citrate affects the expression of these genes at both the transcriptional and post-transcriptional levels.

  10. Microcystin-LR induced developmental toxicity and apoptosis in zebrafish (Danio rerio) larvae by activation of ER stress response.

    Science.gov (United States)

    Qi, Mei; Dang, Yao; Xu, Qinglong; Yu, Liqin; Liu, Chunsheng; Yuan, Yongchao; Wang, Jianghua

    2016-08-01

    Recent studies have demonstrated that cyanobacteria-derived Microcystin-LR (MC-LR) can cause developmental toxicity and trigger apoptosis in zebrafish (Danio rerio) larvae, but the underlying mechanisms remain largely unknown. In this study, we tested the hypothesis that the mechanism by which MC-LR induces developmental toxicity is through activation of endoplasmic reticulum (ER) stress. MC-LR (4.0 μM) exposure through submersion caused serious developmental toxicity, such as malformation, growth delay and decreased heart rates in zebrafish larvae, which could be inhibited by ER stress blocker, tauroursodeoxycholic acid (TUDCA, 20 μM). Meanwhile, acridine orange (AO) staining showed TUDCA could rescue cell apoptosis in heart area in zebrafish larvae resulted by MC-LR exposure. Real-time polymerase chain reaction (real-time PCR) analysis demonstrated that MC-LR induced activation of ER stress which consequently triggered apoptosis in zebrafish larvae. Protein expression examined by western blot indicated that MC-LR could activate MAPK8/Bcl-2/Bax pathway and caspase-dependent apoptotic pathway in zebrafish larva and the effects were mitigated by inhibition of ER stress. Taken together, the results observed in this study suggested that ER stress plays a critical role in developmental toxicity and apoptosis in zebrafish embryos exposed to MC-LR. PMID:27219292

  11. Synthesis, characterizations, photocatalytic and sensing studies of ZnO nanocapsules

    Energy Technology Data Exchange (ETDEWEB)

    Faisal, M., E-mail: mdfaisalahsan@gmail.com [Advanced Materials and Nano-Engineering Laboratory (AMNEL) and Centre for Advanced Materials and Nano-Engineering (CAMNE), Faculty of Science and Arts, Najran University, P. O. Box 1988, Najran, 11001 (Saudi Arabia); Khan, Sher Bahadar [Advanced Materials and Nano-Engineering Laboratory (AMNEL) and Centre for Advanced Materials and Nano-Engineering (CAMNE), Faculty of Science and Arts, Najran University, P. O. Box 1988, Najran, 11001 (Saudi Arabia); Center of Excellence for Advanced Materials Research, King Abdulaziz University, Jeddah 21589, P.O. Box 80203 (Saudi Arabia); Chemistry Department, Faculty of Science, King Abdulaziz University, P. O. Box 80203, Jeddah 21589 (Saudi Arabia); Rahman, Mohammed M.; Jamal, Aslam [Advanced Materials and Nano-Engineering Laboratory (AMNEL) and Centre for Advanced Materials and Nano-Engineering (CAMNE), Faculty of Science and Arts, Najran University, P. O. Box 1988, Najran, 11001 (Saudi Arabia); Asiri, Abdullah M. [Center of Excellence for Advanced Materials Research, King Abdulaziz University, Jeddah 21589, P.O. Box 80203 (Saudi Arabia); Chemistry Department, Faculty of Science, King Abdulaziz University, P. O. Box 80203, Jeddah 21589 (Saudi Arabia); Abdullah, M.M. [Advanced Materials and Nano-Engineering Laboratory (AMNEL) and Centre for Advanced Materials and Nano-Engineering (CAMNE), Faculty of Science and Arts, Najran University, P. O. Box 1988, Najran, 11001 (Saudi Arabia)

    2011-11-01

    ZnO nanocapsules have been synthesized hydrothermally. The structural and morophological properties were investigated using X-ray powder diffraction (XRD), field emission scanning electron microscopy (FESEM), FTIR, Raman, EDS and UV-vis absorption spectroscopy. For the first time chemical sensing properties of the synthesized ZnO nanocapsules have been investigated by I-V technique, where chloroform is used as a target compound. The chloroform sensors show good sensitivity (0.478 {mu}A cm{sup -2} mM{sup -1}), lower detection limit (6.67 {mu}M), and large linear dynamic range (LDR, 12.0 {mu}M-12.0 mM) with good linearity (R, 0.8523) in short response time. Additionally, photocatalytic activity of the prepared capsule shaped ZnO photocatalyst was evaluated by the degradation of acridine orange. Prepared ZnO nanocapsules posses high photocatalytic activity when compared with TiO{sub 2}-UV100.

  12. High-Efficiency Selective Electron Tunnelling in a Heterostructure Photovoltaic Diode.

    Science.gov (United States)

    Jia, Chuancheng; Ma, Wei; Gu, Chunhui; Chen, Hongliang; Yu, Haomiao; Li, Xinxi; Zhang, Fan; Gu, Lin; Xia, Andong; Hou, Xiaoyuan; Meng, Sheng; Guo, Xuefeng

    2016-06-01

    A heterostructure photovoltaic diode featuring an all-solid-state TiO2/graphene/dye ternary interface with high-efficiency photogenerated charge separation/transport is described here. Light absorption is accomplished by dye molecules deposited on the outside surface of graphene as photoreceptors to produce photoexcited electron-hole pairs. Unlike conventional photovoltaic conversion, in this heterostructure both photoexcited electrons and holes tunnel along the same direction into graphene, but only electrons display efficient ballistic transport toward the TiO2 transport layer, thus leading to effective photon-to-electricity conversion. On the basis of this ipsilateral selective electron tunnelling (ISET) mechanism, a model monolayer photovoltaic device (PVD) possessing a TiO2/graphene/acridine orange ternary interface showed ∼86.8% interfacial separation/collection efficiency, which guaranteed an ultrahigh absorbed photon-to-current efficiency (APCE, ∼80%). Such an ISET-based PVD may become a fundamental device architecture for photovoltaic solar cells, photoelectric detectors, and other novel optoelectronic applications with obvious advantages, such as high efficiency, easy fabrication, scalability, and universal availability of cost-effective materials. PMID:27183191

  13. The Use of Lysosomotropic Dyes to Exclude Lysosomal Membrane Permeabilization.

    Science.gov (United States)

    Repnik, Urška; Česen, Maruša Hafner; Turk, Boris

    2016-01-01

    Progressive lowering of pH is characteristic for the endocytic pathway and enables efficient degradation of molecules by hydrolytic enzymes at its distal end. The existence of the proton gradient over the endosomal/lysosomal membranes depends on the action of the vacuolar ATPase (v-ATPase). During lysosomal membrane permeabilization (LMP), protons leak through the destabilized membrane, resulting in loss of the pH gradient. Here, we present a protocol showing how this effect can be detected by staining cells with lysosomotropic dyes, which accumulate in acidic organelles after protonation. During LMP, cells lose the ability to retain these dyes and therefore appear pale. Among the most commonly used lysosomotropic dyes are LysoTracker reagents and acridine orange. Cells can be analyzed with a fluorescence microscope; however, flow-cytometric analysis enables fast, objective, and reliable evaluation of differences between samples. Advantages of the technique include the fact that sample preparation is relatively simple and can be scaled-up to test several different compounds or conditions. However, as we will discuss, cells treated with v-ATPase inhibitors also lose the pH gradient across lysosomal membranes and cannot be stained with lysosomotropic dyes, although this is not accompanied by LMP. Therefore, merely observing loss of staining is not in itself a proof of LMP. PMID:27140914

  14. Growth Inhibition and Apoptosis Inducing Mechanisms of Curcumin on Human Ovarian Cancer Cell Line A2780

    Institute of Scientific and Technical Information of China (English)

    ZHENG Li-duan; TONG Qiang-song; WU Cui-huan

    2006-01-01

    Objective: To explore the growth inhibition effects and apoptosis inducing mechanisms of curcumin on human ovarian cancer cell line A2780. Methods: After treatment with 10-50 μmol/L curcumin for 6-24 h, the growth activity of A2780 cancer cells were studied by [ 4, 5-dimethylthiazol-2-yl]-2, 5-diphenyItetrazolium bromide (MTT) colorimetry. Cellular apoptosis was inspected by flow cytometery and acridine orange-ethidium bromide fluorescent staining methods. The fragmentation of cellular chromosome DNA was detected by DNA ladder, the ultrastructural change was observed under a transmission electron microscope,and the protein levels of nuclear factor-kappa B (NF-κB, P65) and cysteinyl aspartate specific protease-3 (Caspase-3) in ovarian cancer cells were measured by immunohistochemistry. Results: After treatment with various concentrations of curcumin, the growth inhibition rates of cancer cells reached 62.05%- 89.24%,with sub-G1 peaks appearing on histogram. Part of the cancer cells showed characteristic morphological changes of apoptosis under fluorescence and electron microscopes, and the rate of apoptosis was 21.5 % -33.5%. The protein expression of NF-κB was decreased, while that of Caspase-3 was increased in a timedependent manner. Conclusion: Curcumin could significantly inhibit the growth of human ovarian cancer cells;inducing apoptosis through up-regulating Caspase-3 and down-regulating gene expression of NF-κB is probably one of its molecular mechanisms.

  15. A fluorescence-based centrifugal microfluidic system for parallel detection of multiple allergens

    Science.gov (United States)

    Chen, Q. L.; Ho, H. P.; Cheung, K. L.; Kong, S. K.; Suen, Y. K.; Kwan, Y. W.; Li, W. J.; Wong, C. K.

    2010-02-01

    This paper reports a robust polymer based centrifugal microfluidic analysis system that can provide parallel detection of multiple allergens in vitro. Many commercial food products (milk, bean, pollen, etc.) may introduce allergy to people. A low-cost device for rapid detection of allergens is highly desirable. With this as the objective, we have studied the feasibility of using a rotating disk device incorporating centrifugal microfluidics for performing actuationfree and multi-analyte detection of different allergen species with minimum sample usage and fast response time. Degranulation in basophils or mast cells is an indicator to demonstrate allergic reaction. In this connection, we used acridine orange (AO) to demonstrate degranulation in KU812 human basophils. It was found that the AO was released from granules when cells were stimulated by ionomycin, thus signifying the release of histamine which accounts for allergy symptoms [1-2]. Within this rotating optical platform, major microfluidic components including sample reservoirs, reaction chambers, microchannel and flow-control compartments are integrated into a single bio-compatible polydimethylsiloxane (PDMS) substrate. The flow sequence and reaction time can be controlled precisely. Sequentially through varying the spinning speed, the disk may perform a variety of steps on sample loading, reaction and detection. Our work demonstrates the feasibility of using centrifugation as a possible immunoassay system in the future.

  16. Anti-tumor Effects of a Recombinant Fowlpox Virus Expressing Apoptin on Human Cervical Carcinoma in Vivo and in Vitro

    Institute of Scientific and Technical Information of China (English)

    ZHU Ji-hong; JIN Ning-yi; LI Xiao; SUN Li-li; ZHANG Mu-chun; KAN Shi-fu; LIU Lei; HUANG Hai-yan; YANG Guo-hua; PIAO Bing-guo

    2011-01-01

    Apoptin is a chicken anemia virus-derived,p53-independent,bcl-2-insensitive apoptotic protein with the ability to specifically induce apoptosis of various human tumor cells,but not of normal diploid cells.To explore the application of apoptin in tumor gene therapy,we used a recombinant fowlpox virus expressing apoptin protein (vFV-Apoptin) to investigate the anti-tumor effectes of vFV-Apoptin on human cervical carcinoma(HeLa) cells in vivo and in vitro through 3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyltetrazolium bromide(MTT) assay,acridine orage/ethidium bromide(AO/EB) and annexin V staining test,respectively.The results show that vFV-Apoptin inhibites the proliferation of HeLa cells in vitro through inducing the apoptosis of HeLa cells,and the inhibition effect of vFV-Apoptin has a dose-effect and time-effect relationship.The results of animal models show that vFV-Apoptin significantly inhibits tumor growth,extends the lifespan of animals and improves the mean survival.Experimental results indicate that vFV-Apoptin has a potential application in the tumor gene therapy.

  17. Forster Resonance Energy Transfer and Laser Fluorescent Analysis of Defects in DNA Double Helix

    CERN Document Server

    Bregadze, Vasil G; Giorgadze, Tamar G; Jaliashvili, Zaza V; Chkhaberidze, Jemal G; Monaselidze, Jamlet R; Khuskivadze, Temur B

    2013-01-01

    Real time laser induced fluorescence spectroscopy usage for microanalysis of DNA double helix defects is shown. The method is based on Forster resonance energy transfer (FRET) in intercalator-donor pair (acridine orange as a donor and ethidium bromide as an acceptor). Transition metal ions such as Cu(II), Cu(I), Ag(I), silver nanoparticles (AgNPs), photo- and thermo effects were used to cause double helix defects in DNA. FRET radii were experimentally estimated in background electrolyte solution (0.01 M NaNO3) and proved to be 3.9 +- 0.3 nm and the data are in satisfactory agreement with the theoretically calculated value Ro = 3.5 +- 0.3 nm. Concentration of DNA sites, exposed to Cu(II), Cu(I), Ag(I) ions, AgNPs impact as well as laser irradiation ({\\lambda} = 457 nm) and temperature, which are applicable for intercalation, were estimated in relative units. FRET method allows to estimate the concentration of double helix areas with high quality stability applicable for intercalation in DNA after it was subjec...

  18. HERBAL MEDICINE 960 RECIPE REGULATES THE PROLIFERATION AND APOPTOSIS OF HUMAN HEPATOMA CELL LINE SMMC-7721 CELLS

    Institute of Scientific and Technical Information of China (English)

    王昌俊; 李建军; 陈庆强; 陈伟; 钱伯文

    2001-01-01

    To investigate the mechanism of 960 recipe regulating the proliferation and apoptosis of human hepatoma SMMC7721 cells.Methods 960 recipe-containing serum was derived from rats that pre-treated with 960 recipe through gastrogavage, and was added to cultured human hepatoma SMMC-7721 cells while the normal rat serum was tested as control. Cell proliferation was measured with 3H-TdR incorporation. Cell morphology was tested by acridine orange staining. Cell apoptosis and expressions of p53, bcl-2 and p21ras gene protein were analyzed with flow cytometry.Results After treating with 960 recipe, the inhibitory rate of 3H-TdR was 42.2% for 48h. Cell morphology showed typical apoptotic cells with condensed and fragmented nuclei. There were typical apoptotic peaks in DNA histogram, the apoptotic rate being 21.25% and 27.77% for 24h, 48h respectively. Cell cycle analysis showed that cells were arrested at S-phase by treating with 960 recipe for 24h and at G0/G1 phase for 48h. The expression of p53 increased, but bcl-2 and ras were reduced by treating with 960 recipe for 24h.Conclusion 960 recipe can inhibit proliferation and induce apoptosis of human hepatoma cells, and affect the cell cycle, the expression of oncogene and tumor suppressor gene. These might be the main antihepatoma mechanisms of 960 recipe.

  19. Composition of the Extracellular Matrix of Lymphatic Novel Threadlike Structures: Is It Keratin?

    Directory of Open Access Journals (Sweden)

    Hyub Huh

    2013-01-01

    Full Text Available Background. The lumen of novel threadlike structures (NTSs is enclosed by a single layer of endothelial cells surrounded by extracellular matrix (ECM. We hypothesized that collagen may be a component of the ECM associated with lymphatic NTSs. Methods. Six female New Zealand white rabbits were anesthetized, and the NTS structures within lymphatic vessels were identified by contrast-enhanced stereomicroscopy or alcian blue staining. Isolated NTS specimens were stained with acridine orange, YOYO-1, and 1,1′-dioctadecyl-3,3,3′,3′-tetramethylindocarbocyanine perchlorate (DiI. The structural and molecular composition of the ECM was investigated using transmission electron microscopy (TEM, electrospray ionization-mass spectrometry, and proteomic analysis. Results. The lymph vessel wall was stained red by DiI, and rod-shaped nuclei were stained green by YOYO-1. The area surrounding the NTS was also stained red and contained green rod-shaped nuclei. TEM images showed that the NTS consisted of many ECM fibers and the ECM fibers appeared to be ~100 nm in diameter and had narrowly spaced striated bands. Proteomic analysis of the lymphatic NTS-associated ECM identified 4 proteins: keratin 10, cytokeratin 3, cytokeratin 12, and soluble adenylyl cyclase. Conclusion. The TEM study suggested that the lymphatic NTS-associated ECM did not contain collagen. This was confirmed by proteomic analysis, which showed that keratin was the major component of the ECM.

  20. Inhibition of Dual Specific Oncolytic Adenovirus on Esophageal Cancer via Activation of Caspases by a Mitochondrial-dependent Pathway

    Institute of Scientific and Technical Information of China (English)

    SU Jia-qiang; CHI Bao-rong; LI Xiao; LIU Lei; LIU Li-ming; QI Yan-xin; WANG Zhuo-yue; JIN Ning-yi

    2012-01-01

    We investigated the anti-tumor effects of dual cancer specific-oncolytic adenovirus Ad-VP on esophageal cancer(EC).The anti-tumor activity of Ad-VP was compared with that of the control recombinant adenoviruses (Ad-GP,Ad-Apoptin,Ad-EGFP) in human esophageal cancer cell EC-109 and human normal liver cell L02 in vitro.In 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide(MTT) assays,the growth of EC-109 cells was slightly inhibited by Ad-GP.Ad-Apoptin and Ad-EGFP.However,Ad-VP induced a significant cytotoxic effect.Infection of EC-109 cells with Ad-VP resulted in a significant induction of apoptosis of them in vitro,detected by 4′,6-diamidino-2-phenylindole(DAPI) or acridine orange and ethidium bromide staining.The results of Western blot and flow cytometric assay indicate the loss of mitochondrial membrane potential(△ψm),the release of cytochrome c and the activation of caspase-3,6 and 7 in Ad-VP infiected EC-109 cells.In contrast,all these assays show almost no effects of the recombinant adenoviruses on L02 cells.These results demonstrate that the treatment of tumors with Ad-VP selectively inhibits tumor growth and induces apoptosis of esophageal cancer cells.Ad-VP may provide a novel and powerful strategy for cancer gene therapy.

  1. Metformin protects rat hepatocytes against bile acid-induced apoptosis.

    Directory of Open Access Journals (Sweden)

    Titia E Woudenberg-Vrenken

    Full Text Available BACKGROUND: Metformin is used in the treatment of Diabetes Mellitus type II and improves liver function in patients with non-alcoholic fatty liver disease (NAFLD. Metformin activates AMP-activated protein kinase (AMPK, the cellular energy sensor that is sensitive to changes in the AMP/ATP-ratio. AMPK is an inhibitor of mammalian target of rapamycin (mTOR. Both AMPK and mTOR are able to modulate cell death. AIM: To evaluate the effects of metformin on hepatocyte cell death. METHODS: Apoptotic cell death was induced in primary rat hepatocytes using either the bile acid glycochenodeoxycholic acid (GCDCA or TNFα in combination with actinomycin D (actD. AMPK, mTOR and phosphoinositide-3 kinase (PI3K/Akt were inhibited using pharmacological inhibitors. Apoptosis and necrosis were quantified by caspase activation, acridine orange staining and Sytox green staining respectively. RESULTS: Metformin dose-dependently reduces GCDCA-induced apoptosis, even when added 2 hours after GCDCA, without increasing necrotic cell death. Metformin does not protect against TNFα/ActD-induced apoptosis. The protective effect of metformin is dependent on an intact PI3-kinase/Akt pathway, but does not require AMPK/mTOR-signaling. Metformin does not inhibit NF-κB activation. CONCLUSION: Metformin protects against bile acid-induced apoptosis and could be considered in the treatment of chronic liver diseases accompanied by inflammation.

  2. Integrating subpathway analysis to identify candidate agents for hepatocellular carcinoma.

    Science.gov (United States)

    Wang, Jiye; Li, Mi; Wang, Yun; Liu, Xiaoping

    2016-01-01

    Hepatocellular carcinoma (HCC) is the second most common cause of cancer-associated death worldwide, characterized by a high invasiveness and resistance to normal anticancer treatments. The need to develop new therapeutic agents for HCC is urgent. Here, we developed a bioinformatics method to identify potential novel drugs for HCC by integrating HCC-related and drug-affected subpathways. By using the RNA-seq data from the TCGA (The Cancer Genome Atlas) database, we first identified 1,763 differentially expressed genes between HCC and normal samples. Next, we identified 104 significant HCC-related subpathways. We also identified the subpathways associated with small molecular drugs in the CMap database. Finally, by integrating HCC-related and drug-affected subpathways, we identified 40 novel small molecular drugs capable of targeting these HCC-involved subpathways. In addition to previously reported agents (ie, calmidazolium), our method also identified potentially novel agents for targeting HCC. We experimentally verified that one of these novel agents, prenylamine, induced HCC cell apoptosis using 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide, an acridine orange/ethidium bromide stain, and electron microscopy. In addition, we found that prenylamine not only affected several classic apoptosis-related proteins, including Bax, Bcl-2, and cytochrome c, but also increased caspase-3 activity. These candidate small molecular drugs identified by us may provide insights into novel therapeutic approaches for HCC. PMID:27022281

  3. Combinational effects of hexane insoluble fraction of Ficus septica Burm.F.and doxorubicin chemotherapy on T47D breast cancer cells

    Institute of Scientific and Technical Information of China (English)

    Agung; Endro; Nugroho; Adam; Hermawan; Dyaningtyas; Dewi; Pamungkas; Putri; Anindya; Novika; Edy; Meiyanto

    2013-01-01

    Objective:To evaluate the effects of n-hexane insoluble fraction(HIF)of Ficus septica leaves in combination with doxorubicin on cytotoxicity,cell cycle and apoptosis induction of breast cancer T47D cell lines.Methods:The in vitro drugs-stimulated cytotoxic effects were determined using MTT assay.Analysis of cell cycle distribution was performed using flowcytometer and the data was analyzed using ModFit LT 3.0 program.Apoptosis assay was earned out by double staining method using ethydium bromide-acridin orange.The expression of cleaved-poly(ADP-ribose)polymerase(PARP)on T47D cell lines was identified using immunocytochemistry.Results:The combination exhibited higher inhibitory effect on cell growth than the single treatment of doxorubicin in T47D cells.In addition,combination of doxorubicin and HIF increased the incidence of cells undergoing apoptosis.HIF could improve doxorubicin cytotoxic effect by changing the accumulation of cell cycle phase from G2/M to G1 phase.The combination also exhibited upregulation of cleaved-PARP in T47D cells.Conclusions:Based on this results,HIF is potential to be developed as co-chemotherapeutic agent for breast cancer by inducing apoptosis and cell cycle arrest.However,the molecular mechanism need to be explored further.

  4. Protective effects of rilmenidine and AGN 192403 on oxidative cytotoxicity and mitochondrial inhibitor-induced cytotoxicity in astrocytes.

    Science.gov (United States)

    Choi, Dong-Hee; Kim, Dong-Hoon; Park, Yun-Gyu; Chun, Boe-Gwun; Choi, Sang-Hyun

    2002-11-15

    Oxidative stress and mitochondrial dysfunction are important aspects of pathogenesis, particularly in the brain, which is highly dependent on oxygen, and the protection of astrocytes is essential for neuroprotection. In this context, imidazoline drugs have been reported to be neuroprotective. Our recent study showed that imidazoline drugs, including guanabenz, inhibit the naphthazarin-induced oxidative cytotoxicity associated with lysosomal destabilization. We now report on a study into the protective effects of rilmenidine and AGN 192403, which have affinity for imidazoline-1 receptors, on the cytotoxicity induced by naphthazarin and inhibitors of mitochondrial respiration in astrocytes. Cytotoxicity was measured grossly by LDH release and by measuring changes in lysosomal membrane stability and features of mitochondrial membrane permeabilization. Naphthazarin-induced cytotoxicity was evidenced by the ordered development of lysosomal acridine orange relocation, decrease in mitochondrial potential, cytochrome c release, and caspase-9 activation, and was inhibited by guanabenz, rilmenidine, and AGN 192403. Antimycin A and rotenone induced mitochondrial dysfunction primarily, and their cytotoxicities were inhibited only by AGN 192403. Rilmenidine and guanabenz may have a lysosomal stabilizing effect, which underlies their protective effects. AGN 192403 might affect the mitochondrial cell death cascades, and had a novel protective effect on the cytotoxicity associated with mitochondrial dysfunction.

  5. Pro-apoptotic effect of Persea americana var. Hass (avocado) on Jurkat lymphoblastic leukemia cells.

    Science.gov (United States)

    Bonilla-Porras, Angelica R; Salazar-Ospina, Andrea; Jimenez-Del-Rio, Marlene; Pereañez-Jimenez, Andres; Velez-Pardo, Carlos

    2013-11-01

    Abstract Context: Therapy for leukemia has a limited efficacy. There is a need to search for alternative anti-leukemia therapies. Persea americana Mill var. Hass (Lauraceae) is a tropical fruit (avocado) that might be used against cancer. Objective: To investigate whether P. americana induces death in Jurkat lymphoblastic leukemia cells. Materials and methods: Four ethanol extracts (0.1, 0.5, 1, 2 and 5 mg/mL) from avocado fruit (endocarp, whole seed, seed and leaves) were analyzed against Jurkat cells. Hydrogen peroxide generation by oxidation of 2',7'-dichlorodihydrofluorescein diacetate to the fluorescent compound 2',7'-dichlorfluorescein assay, acridine orange/ethidium bromide staining, flow cytometry analysis of annexin-V/7-amino-actinomycin, mitochondrial membrane potential and immunocytochemistry detection of transcription factor p53, caspase-3 and apoptosis-inducing factor (AIF) were evaluated. Results: Endocarp, seed, whole seed, and leaf (0.1 mg/mL) extracts induced significant apoptosis in Jurkat cells (p americana extracts function as a pro-apoptotic compound. Leukemic cells are eliminated through an oxidative stress mechanism. This study contributes to the understanding of the molecular mechanism of the avocado and its therapeutic action on leukemia. PMID:24188375

  6. Antihepatoma Activity of Artocarpus communis Is Higher in Fractions with High Artocarpin Content

    Directory of Open Access Journals (Sweden)

    Cheng-Wei Tzeng

    2014-01-01

    Full Text Available Extracts from natural plants have been used in traditional medicine for many centuries worldwide. Artocarpus communis is one such plant that has been used to treat liver cirrhosis, hypertension, and diabetes. To our knowledge, this study is the first to investigate the antihepatoma activity of A. communis toward HepG2 and PLC/PRF/5 cells and the first to explore the relationship between antihepatoma activity and the active compound artocarpin content in different fractions of A. communis. A. communis methanol extract and fractions induced dose-dependent reduction of tumor cell viability. DNA laddering analysis revealed that A. communis extract and fractions did not induce apoptosis in HepG2 and PLC/PRF/5 cells. Instead, acridine orange staining revealed that A. communis triggered autophagic cell death in a dose-dependent manner. The antihepatoma activity of A. communis is attributable to artocarpin. The fractions with the highest artocarpin content were also the fractions with the highest antihepatoma activity in the following order: dichloromethane fraction > methanol extract > ethyl acetate fraction > n-butanol fraction > n-hexane fraction. Taken together, A. communis showed antihepatoma activity through autophagic cell death. The effect was related to artocarpin content. Artocarpin could be considered an indicator of the anticancer potential of A. communis extract.

  7. [Multiplication of Brucella abortus and production of nitric oxide in two macrophage cell lines of different origin].

    Science.gov (United States)

    Serafino, J; Conde, S; Zabal, O; Samartino, L

    2007-01-01

    Brucella abortus is a bacterium which causes abortions and infertility in cattle and undulant fever in humans. It multiplies intracellularly, evading the mechanisms of cellular death. Nitric oxide (NO) is important in the regulation of the immune response. In the present work, we studied the ability of three B. abortus strains to survive intracellularly in two macrophage cell lines. The bacterial multiplication in both cell lines was determined at two different times in UFC/ ml units. Moreover the inoculated cells were also observed under light-field and fluorescence microscopy stained with Giemsa and acridine orange, respectively. The stain of both cellular lines showed similar results with respect to the UFC/ml determination. The presence of B. abortus was confirmed by electronic microscopy. In both macrophage cell lines inoculated with the rough strain RB51, the multiplication diminished and the level of NO was higher, compared with cells inoculated with smooth strains (S19 and 2308). These results suggest that the absence of O-chain of LPS probably affects the intracellular growth of B. abortus. PMID:18390151

  8. Histopatología cardíaca en los traumatismos torácicos cerrados Myocardial histopathology in the closed chest trauma

    Directory of Open Access Journals (Sweden)

    C. Torres Sánchez

    2002-01-01

    and localized or generalized. We have studied 59 cadavers, 43 males and 16 females, of which 40 died for traumatic causes and other 19 for no traumatic causes. Their ages were between 17 and 90 years old, with an average age of 51,33 years, and the mean postmortem was 13,6 hours (SD 8,7; range 3-59 hours. For this histopathology work, we obtained samples of cardiac tissues from the apex, anterior, lateral and posterior side of the left ventricle, lateral side of the right ventricle and middle side of the interventricle wall, in two different levels of the heart. After fixing and processing of the samples, staining with hematoxylin-eosin and orange acridine were performed. The statistical analysis was carried out using SPSS 10.0. Our results showed the usefulness of hematoxylin-eosin staining for detect the previous myocardial afectation and cronic illness, in the other hand the acridine orange is most usseful to detect ischemic lesions in these locations where the processes of perfusion of the cardiac muscle are going to be more affected acording the previous bibliography.

  9. Concentrations of Polycyclic Aromatic Hydrocarbons (PAHs) and Azaarenes in Runoff from Freshly Applied Coal-Tar-Based Pavement Sealcoat

    Science.gov (United States)

    Mahler, B. J.; Van Metre, P. C.

    2013-12-01

    Coal-tar-based sealcoat (CT-sealcoat) is extensively applied to asphalt parking lots and driveways in the U.S. and Canada. Toxicity to fish and invertebrates of runoff from pavement to which CT-sealcoat has been freshly applied has been reported, but relatively little is known about how concentrations of chemicals in runoff change in the hours to days following sealcoat application. We measured the concentrations of 16 U.S. Environmental Protection Agency Priority Pollutant polycyclic aromatic hydrocarbons (PAHs) and 7 azaarenes in 9 samples of simulated runoff from a coal-tar-sealed test plot collected at increasing intervals from 5 hours to 16 weeks following application. Azaarenes, several of which are common constituents in coal-tar pitch, and their oxidized derivatives, azaarones, are an emerging group of little-studied heterocyclic chemicals. Runoff samples were collected by spraying 25 L of a diluted groundwater to 10 m2 on sealed pavement and retrieving the runoff downgradient where the runoff pooled against spill berms. Unfiltered samples were analyzed by GC/MS following liquid-liquid extraction. In the first sample (t=5 hr), phenanthrene had the highest concentration (130 μg/L) among the 16 PAHs. Concentrations of the lower molecular weight (LMW) PAHs (2 and 3 ring) decreased during the 16 weeks following application, and concentrations of the higher molecular weight (HMW) PAHs (4 to 6 ring) increased, coincident with an increase in the concentration of suspended particulates. In the final sample (t=16 weeks), fluoranthene had the highest concentration (36 μg/L) among the 16 PAHs. Of the azaarenes measured, concentrations of acridine and carbazole (107 and 750 μg/L, respectively) in the initial sample exceeded those of any of the PAHs measured except phenanthrene; acridine and carbazole concentrations decreased over the 5 weeks to <5% of their initial values. Samples of dried sealcoat were analyzed the day of application and 5 weeks later. Samples were

  10. Protection of tetracycline on damage of rat bone mesenchymal stem cells induced by glucocorticosteroid%四环素对糖皮质激素诱导的骨髓间充质干细胞损伤的保护作用

    Institute of Scientific and Technical Information of China (English)

    杨智洋; 张德志; 仲兴; 崔凤荣; 郭涛; 张男; 刘丹平

    2011-01-01

    Objective To investigate the effect of tetracycline on rat bone mesenchymal stem cells (BMSCs) injury indued by glucocorticosteroid. Methods The model of rat BMSCs injury induced by glucocorticosteroid in vitro was estab lished, and then isolately cultured in vitro. The BMSCs of rats were divided into six groups; normal control group, hormone control group, 4, 20, 100, 500 u,g/ml tetracycline group. The injury of BMSCs induced by the glucocorticosteroid was de tected by Acridine orange-ethidium bromide double staining. The cytotoxicity of BMSCs was measured by MTT assay, and then drawing cell growth curve. Staining and analysis of bcl-2, bax epressions were detected by immunohistochemical. Western blot was used to detect the expression of caspase-3. Results Acridine orange-ethidium bromide double staining showed that the cell damage was apoptosis. Tetracycline had protection effect on cell proliferation of BMSCs inhibited by glucocorticosteroid by dose-dependent mode. 100 μg/ml tetracycline could induce the apoptosis of BMSCs by increasing the bcl-2, decreasing bax、caspas-3. Conclusion Tetracycline can protect the injury of BMSCs induced by glucocorticoste roid. This provide some experimental pharmaco-foundation for intervention therapy of early osteonecrosis.%目的 探讨四环素对糖皮质激素诱导大鼠骨髓间充质干细胞(BMSCs)损伤的保护作用.方法 体外建立糖皮质激素诱导大鼠BMSCs损伤模型,分离与体外培养大鼠BMSCs.将大鼠BMSCs分为正常对照组、激素对照组,四环素4、20、100、500 μg/ml浓度组.采用吖啶橙/溴化乙锭双荧光染色法观察四环素对激素诱导的BMSCs凋亡的影响;采用MTT法检测各组BMSCs活性,绘制细胞生长曲线;用免疫组织化学方法对细胞凋亡因子bcl-2、bax进行分析;Western blot测定细胞凋亡因子caspase-3表达.结果 吖啶橙/溴化乙锭染色显示细胞损伤为凋亡.四环素对激素抑制BMSCs的增殖有明显保护作用,

  11. A experimental study on isolation,culture and identification of osteoblasts from neonatal New Zealand rabbit%新西兰乳兔成骨细胞分离、培养与鉴定的实验研究

    Institute of Scientific and Technical Information of China (English)

    刘小荣; 张笠; 王勇平; 张玉娟; 邹传瑛

    2014-01-01

    Objective To investigate the experimental methods of isolation ,culture and identification of osteoblasts from neo-natal New Zealand rabbits in vitro .Methods Two-step enzymatic digestion was adopted to isolate osteoblasts from skull tissue of neonatal New Zealand rabbits to conduct primary cultured .Inverted phase contrast microscope was employed to study the cellular morphology ,acridine orange fluorescent staining was used to detect the cell adhesion function ,methyl thiazolyl tetrazolium(MTT) assay was employed to measure their proliferation ,and Alizarin red and tetracycline staining were used to test their mineralization . Results Primary osteoblasts were successfully obtained .Inverted phase contrast microscopy showed non-adherent cells were round ,while adherent cells were irregular fusiform ,triangular or polygonal .Acridine orange staining showed the nuclei of osteo-blasts green fluorescence ,with good adhesion ability .Good mineralization ability was also demonstrated by tetracycline and alizarin red staining .Osteoblasts possessed good proliferation activity .Conclusion Utilization of two-step enzymatic digestion contributes to getting a lot of osteoblasts with typical morphological features and biological activity in a short time .%目的:探讨新西兰乳兔成骨细胞体外分离、培养及鉴定的实验方法。方法采用二次酶消化法从新西兰乳兔颅骨组织块分离成骨细胞并进行原代培养,采用倒置相差显微镜进行形态学观察,吖啶橙荧光染色检测其黏附功能,采用四甲基偶氮唑盐(MTT)法检测其增殖情况,采用茜素红及四环素染色检测其矿化功能。结果成功获得原代成骨细胞;倒置相差显微镜显示未贴壁的细胞呈圆形,贴壁生长的细胞呈不规则梭形、三角形或多角形;吖啶橙染色可见成骨细胞的细胞核呈绿色荧光,具有良好的黏附能力;茜素红染色及四环素染色均显示其有良好的钙化能

  12. 不同粒径纳米氧化铝对体外培养神经细胞凋亡的影响%In Vitro Study on the Effects of Nano-Aluminum Oxide Particles with Various Sizes on Apoptosis of Neuron Cells

    Institute of Scientific and Technical Information of China (English)

    葛翠翠; 李伟庆; 张勤丽; 牛侨

    2012-01-01

    [目的]体外检测不同粒径纳米氧化铝颗粒对昆明乳鼠神经细胞凋亡的影响.[方法]透射电镜观察纳米氧化铝形态与尺寸,以10 nm、50 nm、10 μm Al2O3同剂量对体外培养的昆明乳鼠进行染毒,运用细胞计数试剂盒( cell counting kit,CCK-8)试剂检测细胞活力,用吖啶橙/溴化乙锭(acridine orange/ethidium bromide,AO/EB)双重染色法观察细胞死亡的形态学,采用Rhodamine123染色方法检测线粒体膜电位变化情况,间接反映早期凋亡变化,应用流式细胞仪测定各染毒组细胞凋亡.[结果]纳米粒子超声后,静置前后相比,电镜下可见纳米粒子均匀分布,且粒径小于100nm符合实验要求;AO/EB染色后,细胞形态学发生明显改变;罗丹明测线粒体膜电位,随着纳米氧化铝粒径的减小,膜电位逐渐降低;应用流式细胞术检测凋亡,随着纳米氧化铝粒径的减小,凋亡率逐渐增加.[结论]氧化铝颗粒能够引起原代培养的神经细胞凋亡,且粒径能够影响细胞凋亡率.%[ Objective ] To observe the apoptosis of cultured neurons from neonatal mice induced by nano-aluminum oxide (Al2O3) particles with different sizes in vitro. [ Methods ] Cultured neurons of neonatal mice were treated with nano-aluminum oxide at same doses but with different sizes. The morphological changes of neurons were observed under transmission electron microscopy with acridine orange (AO)/ethidium bromide (EB) double staining. The cell viability was examined with cell counting kit (CCK-8) assay. The collapses of mitochondrial transmembrane potential (△ψm) were measured using Rhodaminel23 staining to detect the undergoing apoptosis and the apoptosis rates were checked by flow cytometry. [ Results ] After ultrasonic for 10min, the nanoparticles were uniformly dispered with their particle sizes less than 100 nm under an electron microscopy both prior to and after standing. After being exposed to nano-alumina, the morphology of

  13. 不育症患者精子DNA碎片化指数与精子常规参数关系的研究%Research on the relationship between sperm DNA fragmentation and semen parameters of infertile men

    Institute of Scientific and Technical Information of China (English)

    孙超; 徐志鹏; 石亮; 戴玉田

    2011-01-01

    目的:近年研究认为精子DNA碎片化是评价精子质量的重要指标,本研究主要运用吖啶橙实验(acridine orange test,AOT)检测不育症患者精子DNA碎片化指数(DNA fragmentation index,DFI)与精子常规参数之间的关系.方法:收集本院男科门诊376例不育症患者的精液标本,采用改良的Neubauer血细胞计数板人工分析精子密度,计算机辅助精液分析(CASA)分析精子活力.精子形态分析采用Shorr染色法,分析标准采用WHO推荐的Tygerberg严格标准.采用AOT检测精子DNA碎片化程度.结果:DFI与精子密度呈负相关(r=-0.150,P=0.003)、与精子总活动率呈负相关(r=-0.114,P=0.028)、与精子前向运动率无统计学相关性(r=-0.089,P=0.085)、与精子正常形态率呈负相关(r=-0.155,P=0.003).结论:精子DNA碎片化指数与精子的密度、总活动率及精子正常形态率具有负相关性,AOT检测的DFI值为男性不育症的诊疗提供了一个新的指标.%Objective:Sperm DNA fragmentation has been assessed as a positive predictor of fertility potential in recent years.The aim of this study is using the acridine orange test (AOT) to analyze the relationship between sperm DNA fragmentation indexes(DFI)and semen conventional parameters of infertility men.Methods:Semen samples were collected from 376 infertility male patients in our andrology outpatient department.The samples were analyzed by improved Neubauer haemocytometer chamber and computer-aided sperm analysis(CASA) system to obtain concentration and motility parameters.All the samples were stained by Short staining method and the smears were analyzed according to Tygerberg strict criteria recommended by World Health Organization (WHO) to obtain morphology parameters.The DFI as marker of sperm DNA damage determined using the AOT.Results:Significant negative correlations were noted between DFI and sperm concentmtion(r=-0.150,P=0.003),motility(r=-0.114,P=0.028)and morphology(r=-0.155,P=0.003).No

  14. 黄芩提取物联合顺铂对人卵巢癌细胞株SKOV-3凋亡的研究%Study on the effects of extracts from Scutellaria baicalensis combined with cisplatin on apoptosis of human ovarian carcinoma cell line SKOV-3

    Institute of Scientific and Technical Information of China (English)

    李婷婷; 徐红梅; 梁作文; 钟加滕; 何津

    2011-01-01

    Objective: To explore the effects of extracts from Scutellaria baicalensis combined with cisplatin on proliferation and apoptosis of human ovarian carcinoma cell line SKOV - 3.Methods: MTI colorimetry was used to detect the effects of extracts from Scutellaria baicalensis combined with cisplatin on proliferation of human ovarian carcinoma cell line SKOV - 3; acridine orange staining was used to observe the apoptosis of human ovarian carcinoma cell line SKOV - 3; flow cytometry was used to analyze cell cycle and apoptosis; the average apoptosis rate was calculated; Western blot was used to observe the expressions of p53, ERK1/2 and p -STAT3.Results: After treated with 300μg/ml extracts from Scutellaria baicalensis and 5μg/ml cisplatin for 48 hours, the inhibition rate of humah ovarian carcinoma cell line SKOV -3 increased from 15.8% (treated with cisplatin only) to 37.3%; acridine orange staining showed apoptotic cells, the apoptosis rate was similar to that of 10μg/ml cisplatin group; the results of flow cytometry showed that the apoptosis rate increased from 15.1% to 42.2% (P <0.05); the results of Western blot showed that the expression levels of p53, ERK1/2 and p - STAT3 were significantly lower than those in 5μg/ml cisplatin group (P <0.05 ).Conclusion: The extracts from Scutellaria baicalensis combined with cisplatin can enhance the apoptotic effect on human ovarian carcinoma cell line SKOV -3, reduce toxicity and improve clinical efficacy.%目的:探讨黄芩提取物(Scutellaria Baicalensis)与顺铂(cisplatin,DDP)联合应用对卵巢癌细胞系SKOV-3增殖及凋亡的影响.方法:采用MTT比色法检测S.Baicalensis与DDP联合应用对SKOV-3细胞增殖的影响,吖啶橙染色观察诱导细胞凋亡情况,流式细胞仪分析细胞周期及凋亡并计算细胞平均凋亡率,Westernblot技术观察细胞内p53、ERK1/2、P-STAT3的表达.结果:300μ/ml S.Baicalensis联合5 μ/ml DDP给药:①48 h后可使肿瘤细

  15. Der Einfluß verschiedener Gefriermethoden bezüglich Morpholigie und Chromatinintegrität auf menschliche Spermatozoen von fertilen und subfertilen Männern

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    Hammadeh ME

    2000-01-01

    Full Text Available Einige Autoren haben gezeigt, daß die computergesteuerte Einfriermethode die Spermienqualität besser aufrechterhält als die Stickstoffdampf-Methode. Andere haben keinen Vorteil festgestellt, zumindest bei menschlichen Spermien. Das Ziel dieser Arbeit war es, zum einen negative Einflüsse der Einfrier-Auftau-Prozedur festzustellen (Kryoschädigung, zum anderen herauszufinden, ob es einen Unterschied zwischen der computergesteuerten Einfrier-Auftau-Technik und der Stickstoffdampf-Technik in Hinblick auf Chromatin- und Morphologieveränderungen sowohl von fertilen als auch von subfertilen Männern gibt. Die Studie umfaßt 37 Spermaproben, 22 nachweislich fertiler Spender (Kontrollgruppe, G2 und 15 Proben von Patienten mit Fertilitätsstörungen (G1, beurteilt nach WHO Richtlinien (1992. Die Proben wurden mit dem Kryoprotektivum Glycerol (HSPM 1:1 gemischt und jeweils in Stickstoffdampf und mit einer biologischen Friermaschine (Planer Serie 10 eingefroren. Vor und nach dem Einfrier-Auftau-Vorgang wurden mehrere Ausstriche angefertigt, um die Morphologie (strict criteria, Kruger 1986 und die Chromatinkondensation mittels Acridine-Orange-Färbung auszuwerten. Der Anteil an kondensiertem Chromatin im nativen Sperma der fertilen Probanden zeigte einen signifikanten Abfall (p 0,001 von 94,1 % ± 7,0 auf 89,1 % ± 8,1 nach Einfrieren mit der biologischen Friermaschine und auf 89,1 % ± 7,7 nach Einfrieren mit Stickstoffdampf. Die entsprechenden Werte der subfertilen Probanden zeigten die gleiche Tendenz und fielen ebenso signifikant (p = 0,001 von 80,4 % ± 9,9 auf 72,7 % ± 10,1 bzw. 62,7 ± 10,1 ab. Die Morphologie der Spermien zeigte die gleich Tendenz: Die Anzahl der morphologisch normalen Spermien (27,9 % ± 6,5, eingefroren mit der biologischen Friermaschine, zeigte in der fertilen Gruppe einen signifikanten Abfall (p = 0,001 auf 21,1 % ± 4,7 (mittlerer prozentualer Abfall auf 65 % und nach Einfrieren mit Stickstoffdampf auf 20,2 % ± 4,7. In

  16. Oxidative desulfurization and denitrogenation of a light gas oil using an oxidation/adsorption continuous flow process

    Energy Technology Data Exchange (ETDEWEB)

    Ishihara, Atsushi; Wang, Danhong; Dumeignil, Franck; Amano, Hiroshi; Qian, Eika Weihua; Kabe, Toshiaki [Department of Chemical Engineering, Tokyo University of Agriculture and Technology, 2-24-16 Nakacho, Koganei, Tokyo 184-8588 (Japan)

    2005-01-28

    The oxidation of undesirable sulfur compounds present in a desulfurized light gas oil (LGO; sulfur content: 39ppm) was performed with tert-butyl hydroperoxide (t-BuOOH) as the oxidant in the presence of a 16wt.% MoO{sub 3}/Al{sub 2}O{sub 3} catalyst. The oxidation activity of the sulfur compounds in the light gas oil increased when the O/S molar ratio increased up to 15; the activity slightly decreased for higher ratios. This optimal ratio was significantly higher than the stoichiometric one (=2) due to parallel oxidation reactions of olefins, etc., in the LGO. Further, we compared the oxidation reactivity of dibenzothiophene (DBT), 4,6-dimethyldibenzothiophene (4,6-DMDBT), and trimethyldibenzothiophene (C{sub 3}-DBT), which are refractory compounds present in the light gas oil. The reactivity decreased in the order DBT|4,6-DMDBT>C{sub 3}-DBT, irrespective of the WHSV and the temperature. Subsequent mathematical treatment revealed that the oxidative reaction of each sulfur compound follows a first-order kinetics. We found an activation energy of 32+/-2kJmol{sup -1}, whatever the compound, suggesting that the oxidation mechanism was the same for these compounds. Then, according to the proposed global process, the previously oxidized molecules in the treated light gas oil were further removed by adsorption over a silica gel at ambient temperature. As a result, the total sulfur content could be decreased after oxidation/adsorption to less than 5ppm. Further, N-containing model compounds were also treated according to the same procedure and the denitrogenation performance decreased in the order indole>quinoline>acridine>carbazole. Subsequently, the same process allowed decreasing the N content in the LGO from an initial value of 13.5ppm to a value of 0.8ppm, which is a remarkable result.

  17. Gametocytocidal screen identifies novel chemical classes with Plasmodium falciparum transmission blocking activity.

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    Natalie G Sanders

    Full Text Available Discovery of transmission blocking compounds is an important intervention strategy necessary to eliminate and eradicate malaria. To date only a small number of drugs that inhibit gametocyte development and thereby transmission from the mosquito to the human host exist. This limitation is largely due to a lack of screening assays easily adaptable to high throughput because of multiple incubation steps or the requirement for high gametocytemia. Here we report the discovery of new compounds with gametocytocidal activity using a simple and robust SYBR Green I- based DNA assay. Our assay utilizes the exflagellation step in male gametocytes and a background suppressor, which masks the staining of dead cells to achieve healthy signal to noise ratio by increasing signal of viable parasites and subtracting signal from dead parasites. By determining the contribution of exflagellation to fluorescent signal and using appropriate cutoff values, we were able to screen for gametocytocidal compounds. After assay validation and optimization, we screened an FDA approved drug library of approximately 1500 compounds, as well as the 400 compound MMV malaria box and identified 44 gametocytocidal compounds with sub to low micromolar IC50s. Major classes of compounds with gametocytocidal activity included quaternary ammonium compounds with structural similarity to choline, acridine-like compounds similar to quinacrine and pyronaridine, as well as antidepressant, antineoplastic, and anthelminthic compounds. Top drug candidates showed near complete transmission blocking in membrane feeding assays. This assay is simple, reproducible and demonstrated robust Z-factor values at low gametocytemia levels, making it amenable to HTS for identification of novel and potent gametocytocidal compounds.

  18. Anti-pancreatic cancer deliverables from sea: first-hand evidence on the efficacy, molecular targets and mode of action for multifarious polyphenols from five different brown-algae.

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    Sheeja Aravindan

    Full Text Available Pancreatic cancer (PC remains the fourth leading cause of cancer death with an unacceptable survival that has remained relatively unchanged over the past 25 years. The presence of occult or clinical metastases at the time of diagnosis together with the lack of effective chemotherapies pose a dire need for designing new and targeted therapeutic deliverables that favors the clinical outcome. Herein, we investigated the anti-tumorigenic potential of polyphenols from five different brown-algae in human PC cells (MiaPaCa-2, Panc-1, BXPC-3 and Panc-3.27. Total anti-oxidant capacity (TAC analysis on stepwise polyphenol separations with increasing polarity (Hexane-DCM-EA-methanol identified high levels of TAC in DCM and EA extractions across all seaweeds assessed. All DCM and EA separated polyphenols induced a dose-dependent and sustained (time-independent inhibition of cell proliferation and viability. Further, these polyphenols profoundly enhanced DNA damage (acridine orange/Ethidium bromide staining and DNA fragmentation in all the cell lines investigated. More importantly, luciferase reporter assay revealed a significant inhibition of NFκB transcription in cells treated with polyphenols. Interestingly, QPCR analysis identified a differential yet definite regulation of pro-tumorigenic EGFR, VEGFA, AKT, hTERT, kRas, Bcl2, FGFα and PDGFα transcription in cells treated with DCM and EA polyphenols. Immunoblotting validates the inhibitory potential of seaweed polyphenols in EGFR phosphorylation, kRas, AurKβ and Stat3. Together, these data suggest that intermediate polarity based fractions of seaweed polyphenols may significantly potentiate tumor cell killing and may serve as potential drug deliverable for PC cure. More Studies dissecting out the active constituents in potent fractions, mechanisms of action and synergism, if any, are warranted and are currently in process.

  19. TP53inp1 Gene Is Implicated in Early Radiation Response in Human Fibroblast Cells

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    Nikolett Sándor

    2015-10-01

    Full Text Available Tumor protein 53-induced nuclear protein-1 (TP53inp1 is expressed by activation via p53 and p73. The purpose of our study was to investigate the role of TP53inp1 in response of fibroblasts to ionizing radiation. γ-Ray radiation dose-dependently induces the expression of TP53inp1 in human immortalized fibroblast (F11hT cells. Stable silencing of TP53inp1 was done via lentiviral transfection of shRNA in F11hT cells. After irradiation the clonogenic survival of TP53inp1 knockdown (F11hT-shTP cells was compared to cells transfected with non-targeting (NT shRNA. Radiation-induced senescence was measured by SA-β-Gal staining and autophagy was detected by Acridine Orange dye and microtubule-associated protein-1 light chain 3 (LC3B immunostaining. The expression of TP53inp1, GDF-15, and CDKN1A and alterations in radiation induced mitochondrial DNA deletions were evaluated by qPCR. TP53inp1 was required for radiation (IR induced maximal elevation of CDKN1A and GDF-15 expressions. Mitochondrial DNA deletions were increased and autophagy was deregulated following irradiation in the absence of TP53inp1. Finally, we showed that silencing of TP53inp1 enhances the radiation sensitivity of fibroblast cells. These data suggest functional roles for TP53inp1 in radiation-induced autophagy and survival. Taken together, we suppose that silencing of TP53inp1 leads radiation induced autophagy impairment and induces accumulation of damaged mitochondria in primary human fibroblasts.

  20. Synthesis, characterization and anticancer activity studies of ruthenium(II) polypyridyl complexes on A549 cells.

    Science.gov (United States)

    Zeng, Chuan-Chuan; Jiang, Guang-Bin; Lai, Shang-Hai; Zhang, Cheng; Yin, Hui; Tang, Bing; Wan, Dan; Liu, Yun-Jun

    2016-08-01

    Four new ruthenium(II) polypyridyl complexes [Ru(N-N)2(bddp)](ClO4)21-4 (N-N=dmb: 4,4'-dimethyl-2,2'-bipyridine 1, bpy: 2,2'-bipyridine 2, phen: 1,10-phenanthroline 3 and dmp: 2,9-dimethyl-1,10-phenanthroline 4, bddp=benzilo[2,3-b]-1,4-diazabenzo[i]dipyrido[3,2-a:2',3'-c]phenazine) were synthesized and characterized by elemental analysis, ESI-MS and (1)H NMR. The cytotoxicity in vitro of the complexes against BEL-7402, HeLa, MG-63 and A549 cell lines was investigated by MTT method. The complexes show high cytotoxic activity toward the selected cell lines with an IC50 value ranging from 5.3±0.6 to 15.7±3.6μM. The apoptosis was studied with acridine orange (AO)/ethdium bromide (EB) and Hoechst 33258 staining methods. The cellular uptake was investigated with DAPI staining method. The reactive oxygen species (ROS) and mitochondrial membrane potential were performed under fluorescent microscope and flow cytometry. The complexes can induce an increase in the ROS levels and a decrease in the mitochondrial membrane potential. The comet assay was studied with fluorescent microscope. The percentage in apoptotic and necrotic cells and cell cycle arrest were assayed by flow cytometry. The effects of the complexes on the expression of caspases and Bcl-2 family proteins were studied by western blot analysis. The results show that the complexes induce apoptosis in A549 cells through an ROS-mediated mitochondrial dysfunction pathway, which was accompanied by regulating the expression of Bcl-2 family proteins. PMID:27288660

  1. Polyphenol stabilized colloidal gold nanoparticles from Abutilon indicum leaf extract induce apoptosis in HT-29 colon cancer cells.

    Science.gov (United States)

    Mata, Rani; Nakkala, Jayachandra Reddy; Sadras, Sudha Rani

    2016-07-01

    Green synthesized gold nanoparticles have received substantial attention owing to their biomedical applications, particularly in cancer therapy. Although anticancer activities of green synthesized gold nanoparticles have been reported earlier, the underlying mechanism behind their anticancer activity is still to be understood. The present study, describes the green synthesis of Abutilon indicum gold nanoparticles (AIGNPs) from Abutilon indicum leaf extract (AILE) and their cytotoxic mechanism in colon cancer cells. Dimensions of spherical shaped AIGNPs were found to be in the range of 1-20nm as determined by TEM. GC-MS and FTIR analysis indicated the presence of polyphenolic groups in AILE, which might have been involved in the stabilization of AIGNPs. In vitro free radical scavenging analysis revealed the radical quenching activity of AIGNPs. Further, the AIGNPs exhibited cytotoxicity in HT-29 colon cancer cells with IC50 values of 210 and 180μg/mL after 24 and 48h. This was mediated through nuclear morphological changes and cell membrane damage as evidenced by acridine orange/ethidium bromide, propidium iodide and AnnexinV-Cy3 staining methods. Mechanism of the observed cytotoxicity of AIGNPs was explained on the basis of increased levels of reactive oxygen species and simultaneous reduction in cellular antioxidants, which might have caused mitochondrial membrane potential loss, DNA damage and G1/S phase cell cycle arrest. Expression of cleaved Caspase-9, Caspase-8, Caspase-3, Lamin A/C and PARP, provided the clues for the induction of intrinsic and extrinsic apoptosis pathways in AIGNPs treated HT-29 cells. The study provides a preliminary guidance towards the development of colon cancer therapy using green synthesized gold nanoparticles. PMID:27038915

  2. Usefulness of quantitative buffy coat blood parasite detection system in diagnosis of malaria

    Directory of Open Access Journals (Sweden)

    Pinto M

    2001-01-01

    Full Text Available A rapid test for diagnosis of malaria based on acridine orange staining of centrifuged blood samples in a microhematocrit tube (QBC was compared with thick and thin peripheral blood smears in 2274 samples. Malaria was diagnosed in 239 (10.5% patients by Leishman′s staining technique and QBC method. The QBC method allowed detection of an additional 89 (3.9% cases. Thus the prevalence rate of malaria during the study was 14.4%. In 1946 patients who were negative by the QBC technique, the Leishman′s stained smears did not provide any help in malaria diagnosis. Analysis of the relative quantity of parasites in the specimens, in the QBC method, revealed that 80 out of 89 QBC positive but smear negative cases, had a very low parasite number (less than 10 parasites per QBC field. Although QBC method was superior to the smear for malarial parasite detection, species identification was not possible in 26 (7.9% cases by this technique. In 95.7% (n = 314 QBC positive cases, the buffy coat in the QBC tube appeared pigmented (gray to black. The colour of the buffy coat was therefore considered by us as a predictor of positivity and could be taken as an indicator for a careful and more prolonged search for the parasites. Thus, the QBC technique has its advantages in terms of speed, sensitivity and ease, especially in an endemic area as ours, where the level of parasitaemia is low and more than 70 to 80 smears need to be examined per day. However, the age old Romanowsky stains still appear superior for species identification.

  3. Application of UV-Vis spectrophotometric process for the assessment of indoloacridines as free radical scavenger.

    Science.gov (United States)

    Sridharan, Makuteswaran; Prasad, K J Rajendra; Madhumitha, G; Al-Dhabi, Naif Abdullah; Arasu, Mariadhas Valan

    2016-09-01

    A conventional approach has been used to synthesis Indole fused acridine, 4a-e. In this paper to achieve the target molecule, 4 the reaction was performed via two steps. In step 1, there was a reaction between Carbazolone, 1 and benzophenone, 2 to get dihydroindoloacridine, 3. In step 2, compound, 3 was treated with 5% Palladium/Carbon in the presence of diphenyl ether for 5h to give a dark brown product, 4. The column chromatography was used to purify final product, 4. All the synthesized compounds such as 3 and 4 were characterized by melting point, FTIR, (1)H NMR, and Mass spectra. Further to check the purity of the compounds it was subjected to CHN analyzer. The target molecules such as 3 and 4 were screened for antimicrobial studies against bacteria such as Bacillus subtilis (B. subtilis), Staphylococcus aureus (S. aureus), Klebsiella pneumonia (K. pneumonia), Salmonella typhi (S. typhi); and fungi like Aspergillus niger (A. niger), Aspergillus fumigatus (A. fumigatus). The obtained results clearly proves that the target molecules shown reasonable activity against K. pneumonia and A. niger. Further the compounds were screened for free radical scavenging activity using 2,2-diphenyl-1-picrylhydrazyl (DPPH). The free radical scavenging property was performed using UV-Visible spectroscopy. The results were compared with the standard BHT (Butylated Hydroxy Toluene). Compounds, 4a and 4e were shown higher percentage of inhibition when compare to the standard. The result confirms that further research on indoloacridine will leads effective drug to the market. PMID:27491030

  4. HPLC detection of loss rate and cell migration of HUVECs in a proanthocyanidin cross-linked recombinant human collagen-peptide (RHC)–chitosan scaffold

    Energy Technology Data Exchange (ETDEWEB)

    Zhang, Jing; Deng, Aipeng [School of Environmental and Biological Engineering, Nanjing University of Science and Technology, Nanjing 210094 (China); Yang, Yang [Faculty of Engineering, University of Nottingham, Nottingham NG7 2RD (United Kingdom); Gao, Lihu; Xu, Na; Liu, Xin; Hu, Lunxiang; Chen, Junhua [School of Environmental and Biological Engineering, Nanjing University of Science and Technology, Nanjing 210094 (China); Yang, Shulin, E-mail: yshulin@njust.edu.cn [School of Environmental and Biological Engineering, Nanjing University of Science and Technology, Nanjing 210094 (China)

    2015-11-01

    Porous scaffolds with appropriate pore structure, biocompatibility, mechanical property and processability play an important role in tissue engineering. In this paper, we fabricated a recombinant human collagen-peptide (RHC)–chitosan scaffold cross-linked by premixing 30% proanthocyanidin (PA) in one-step freeze-drying. To remove the residual acetic acid, optimized 0.2 M phosphate buffer of pH 6.24 with 30% ethanol (PBSE) was selected to neutralize the lyophilized scaffold followed by three times deionized water rinse. Ninhydrin assay was used to characterize the components loss during the fabrication process. To detect the exact RHC loss under optimized neutralization condition, high performance liquid chromatography (HPLC) equipped size exclusion chromatography column was used and the total RHC loss rate through PBSE rinse was 19.5 ± 5.08%. Fourier transform infrared spectroscopy (FT-IR) indicated hydrogen bonding among RHC, chitosan and PA, it also presented a probative but not strong hydrophobic interaction between phenyl rings of polyphenols and pyrrolidine rings of proline in RHC. Further, human umbilical vein endothelial cell (HUVEC) viability analyzed by a scanning electron microscope (SEM) and acridine orange/ethidium bromide (AO/EB) fluorescence staining exhibited that this scaffold could not only promote cell proliferation on scaffold surface but also permit cells migration into the scaffold. qRT-PCR exhibited that the optimized scaffold could stimulate angiogenesis associated genes VEGF and CD31 expression. These characterizations indicated that this scaffold can be considered as an ideal candidate for tissue engineering. - Highlights: • PA cross-linked recombinant human collagen–chitosan scaffold. • Fabrication in one-step lyophilization with neutralization. • HPLC detection of RHC loss rate • HUVEC proliferation and migration in scaffold • Angiogenesis associated gene expressions were increased in scaffold cell culturing.

  5. Influence of Cu content on the cell biocompatibility of Ti–Cu sintered alloys

    Energy Technology Data Exchange (ETDEWEB)

    Zhang, Erlin, E-mail: zhangel@atm.neu.edu.cn [Key Lab. for Anisotropy and Texture of Materials, Education Ministry of China, Northeastern University, Shenyang 110819 (China); Jiamusi University, Jiamusi 154007 (China); Zheng, Lanlan [Jiamusi University, Jiamusi 154007 (China); Liu, Jie [Key Lab. for Anisotropy and Texture of Materials, Education Ministry of China, Northeastern University, Shenyang 110819 (China); Dept. of Prosthodontics, The Affiliated Hospital of Medical College, Qingdao University, Qingdao 266003 (China); Bai, Bing [Dept. of Prosthodontics, School of Stomatology, China Medical University, Liaoning Institute of Dental Research, Shenyang 110001 (China); Liu, Cong [Jiamusi University, Jiamusi 154007 (China)

    2015-01-01

    The cell toxicity and the cell function of Ti–Cu sintered alloys with different Cu contents (2, 5, 10 and 25 wt.%, respectively) have been investigated in comparison with commercial pure titanium in order to assess the influence of Cu content on the cell biocompatibility of the Ti–Cu alloys. The cytotoxicity was studied by examining the MG63 cell response by CCK8 assessment. The cell morphology was evaluated by acridine orange/ethidium bromide (AO/EB) fluorescence and observed under scanning electronic microscopy (SEM). The cell function was monitored by measuring the AKP activity. It has been shown by the AO/EB morphology results that the cell death on both cp-Ti sample and Ti–Cu samples is due to apoptosis rather than necrosis. Although more apoptotic cells were found on the Ti–2Cu and Ti–5Cu samples, no evidence of Cu content dependent manner of apoptosis has been found. SEM observation indicated very good cell adhesion and spread on the cp-Ti sample and the Ti–Cu samples with different Cu contents. CCK8 results displayed that increase in the Cu content in Ti–Cu alloys does not bring about any difference in the cell viability. In addition, AKP test results indicated that no difference in the differentiation of MG63 was found between the cp-Ti and the Ti–Cu samples and among the Ti–Cu samples. All results indicated that Ti–Cu alloys exhibit very good cell biocompatibility and the Cu content up to 25 wt.% in the Ti–Cu alloys has no influence on the cell proliferation and differentiation. - Highlights: • The effect of Cu content on the cell biocompatibility has been investigated. • Cu content shows no influence on the cell proliferation. • Cu content shows no effect on the cell differentiation.

  6. Ethylenediamine functionalized-single-walled nanotube (f-SWNT-assisted in vitro delivery of the oncogene suppressor p53 gene to breast cancer MCF-7 cells

    Directory of Open Access Journals (Sweden)

    Karmakar A

    2011-05-01

    Full Text Available Alokita Karmakar2, Stacie M Bratton1, Enkeleda Dervishi2, Anindya Ghosh3, Meena Mahmood2, Yang Xu2, Lamya Mohammed Saeed2, Thikra Mustafa2, Dan Casciano2, Anna Radominska-Pandya1, Alexandru S Biris21Biochemistry Department, University of Arkansas for Medical Sciences; 2Nanotechnology Center, Applied Science Department; 3Department of Chemistry, University of Arkansas, Little Rock, AR, USAAbstract: A gene delivery concept based on ethylenediamine-functionalized single-walled carbon nanotubes (f-SWCNTs using the oncogene suppressor p53 gene as a model gene was successfully tested in vitro in MCF-7 breast cancer cells. The f-SWCNTs-p53 complexes were introduced into the cell medium at a concentration of 20 µg mL-1 and cells were exposed for 24, 48, and 72 hours. Standard ethidium bromide and acridine orange assays were used to detect apoptotic cells and indicated that a significantly larger percentage of the cells (approx 40% were dead after 72 hours of exposure to f-SWCNTs-p53 as compared to the control cells, which were exposed to only p53 or f-SWCNTs, respectively. To further support the uptake and expression of the genes within the cells, green fluorescent protein-tagged p53, attached to the f-SWCNTs was added to the medium and the complex was observed to be strongly expressed in the cells. Moreover, caspase 3 activity was found to be highly enhanced in cells incubated with the f-SWCNTs-p53 complex, indicating strongly induced apoptosis. This system could be the foundation for novel gene delivery platforms based on the unique structural and morphological properties of multi-functional nanomaterials.Keywords: carbon nanotubes, gene delivery, cancer cells, p53 oncogene suppressor

  7. Toxic Potential of Synthesized Graphene Zinc Oxide Nanocomposite in the Third Instar Larvae of Transgenic Drosophila melanogaster (hsp70-lacZBg9

    Directory of Open Access Journals (Sweden)

    Yasir Hasan Siddique

    2014-01-01

    Full Text Available In the present study the graphene zinc oxide nanocomposite (GZNC was synthesized, characterized, and evaluated for its toxic potential on third instar larvae of transgenic Drosophila melanogaster (hsp70-lacZBg9. The synthesized GZNC was characterized by X-ray diffraction (XRD, Fourier transform infrared spectroscopy (FTIR, thermogravimetric analysis (TGA, scanning electron microscopy (SEM, and transmission electron microscopy (TEM. The GZNC in 0.1% dimethyl sulphoxide (DMSO was sonicated for 10 minutes and the final concentrations 0.033, 0.099, 0.199, and 3.996 μg/μL of diet were established. The third instar larvae were allowed to feed on it separately for 24 and 48 hr. The hsp70 expression was measured by o-nitrophenyl-β-D-galactopyranoside assay, tissue damage was measured by trypan blue exclusion test, and β-galactosidase activity was monitored by in situ histochemical β-galactosidase staining. Oxidative stress was monitored by performing lipid peroxidation assay and total protein estimation. Ethidium bromide/acridine orange staining was performed on midgut cells for apoptotic index and the comet assay was performed for the DNA damage. The results of the present study showed that the exposure of 0.199 and 3.996 μg/μL of GZNC was toxic for both 24 hr and 48 hr of exposure. The doses of 0.033 μg/μL and 0.099 of GZNC showed no toxic effects on its exposure to the third instar larvae for 24 hr as well as 48 hr of duration.

  8. Cytotoxicity and DNA Damage Effect of TGA-capped CdTe Quantum Dots

    Institute of Scientific and Technical Information of China (English)

    LI Yan-bo; ZHANG Hai-xia; GUO Cai-xia; HU Gui-qin; DU Hai-ying; JIN Ming-hua; HUANG Pei-li; SUN Zhi-wei; YANG Wen-sheng

    2012-01-01

    The cytotoxicity and DNA damage caused by thioglycolic acid(TGA)-capped cadmium telluride(CdTe)quantum dots(QDs)to hepatocyte line HL-7702 were investigated.Cell viability was measured by 3-(4,5-dimethyl-thiazol-2-yl)-2,5-diphenyltetrazolium bromide(MTT)assay; DNA damage was detected by single cell gel electrophoresis(SCGE); the change of cell cycle progression was examined by propidium iodide(PI)-flow cytometry(FCM);apoptosis was measured by acridine orange/ethidium bromide(AO/EB)assay and Annexin V-FITC/PI-FCM(FITC:fluorescein isothiocyanate).The results show that the cytotoxicity induced by CdTe QDs was increased in a dose-dependent and time-dependent manner; after exposure to QDs for 24 h,as the exposure dose increased,the rate of DNA damage was significantly increased(P<0.05),and the degree of DNA damage was elevated.As the dose of CdTe QDs increased,the percentage of G0/G1 phase cells was significantly decreased(P<0.001),while the percenttages of S and G2/M phases cells were significantly increased(P<0.001).In AO/EB assay,apoptotic cells could be observed under a fluorescence microscope,and apoptotic rate was increased as exposure dose increased.In Annexin V-FITC/PI-FCM assay,the apoptotic rates of CdTe QDs treated groups were significantly increased compared with that of control group(P<0.05).Our studies indicate that CdTe QDs could influence cell viability,and induce DNA damage,the S and G2/M phases arrest and apoptosis of HL-7702.

  9. Fourier transform infrared spectroscopic analysis of sperm chromatin structure and DNA stability.

    Science.gov (United States)

    Oldenhof, H; Schütze, S; Wolkers, W F; Sieme, H

    2016-05-01

    Sperm chromatin structure and condensation determine accessibility for damage, and hence success of fertilization and development. The aim of this study was to reveal characteristic spectral features coinciding with abnormal sperm chromatin packing (i.e., DNA-protein interactions) and decreased fertility, using Fourier transform infrared spectroscopy. Chromatin structure in spermatozoa obtained from different stallions was investigated. Furthermore, spermatozoa were exposed to oxidative stress, or treated with thiol-oxidizing and disulfide-reducing agents, to alter chromatin structure and packing. Spectroscopic studies were corroborated with flow cytometric analyses using the DNA-intercalating fluorescent dye acridine orange. Decreased fertility of individuals correlated with increased abnormal sperm morphology and decreased stability toward induced DNA damage. Treatment with the disulfide reducing agent dithiothreitol resulted in increased sperm chromatin decondensation and DNA accessibility, similar as found for less mature epididymal spermatozoa. In situ infrared spectroscopic analysis revealed that characteristic bands arising from the DNA backbone (ν1230, ν1086, ν1051 cm(-1) ) changed in response to induced oxidative damage, water removal, and decondensation. This coincided with changes in the amide-I region (intensity at ν1620 vs. ν1640 cm(-1) ) denoting concomitant changes in protein secondary structure. Reduction in protein disulfide bonds resulted in a decreased value of the asymmetric to symmetric phosphate band intensity (ν1230/ν1086 cm(-1) ), suggesting that this band ratio is sensitive for the degree of chromatin condensation. Moreover, when analyzing spermatozoa from different individuals, it was found that the asymmetric/symmetric phosphate band ratio negatively correlated with the percentage of morphologically abnormal spermatozoa. PMID:26916383

  10. Laboratory diagnosis of malaria -- overview.

    Science.gov (United States)

    Bhatt, K M

    1994-01-01

    Features of the laboratory diagnosis of malaria are described. Microscope equipment is absolutely essential. Clinical symptoms are inadequate for the proper diagnosis of malaria. Screening for malaria involves identification of all cases where high fever is present in endemic areas. Diagnosis is complicated because many people take antimalarial drugs which reduce the chances of detecting malarial parasites. Confirmation should be made before treatment is administered. A thick blood slide can be quickly and cheaply taken without much training of health personnel. The disadvantage of thick stains is the difficulty in identifying "plasmodium" strains. When a thin smear with Giemsa and Leishmanin stain is used, a light infection may be missed. Thin smears require trained personnel and time, which in peak seasons may be impractical. Urinary tract and viral infections may be confused with malaria. Evidence of parasites can be discerned from thick stains. Modern assay techniques are also available. There are enzyme linked immunosorbent assays (ELISA) and immunofluorescent assay techniques (IFAT), which are frequently used in large scale seroepidemiological studies. DNA probes have the limitation of radioisotope handling problems. Acridine orange fluorescent microscopy with capillary centrifuged blood is a technique which improves the viability of Giemsa stain procedures. This technique is desirable because of the sensitivity and speed of diagnosis. The quantitative buddy coat (GBC) technique is superior to Giemsa stained thick blood film in identifying malaria, but it is not reliable with mixed infections. Advanced techniques are not readily available in local settings. The recommendation is to continue use of thick or thin blood film and trained health personnel. Laboratory results must be interpreted in the context of when the flood film was prepared, prior drug administration, and clinical manifestations.

  11. Potential use of porous titanium-niobium alloy in orthopedic implants: preparation and experimental study of its biocompatibility in vitro.

    Directory of Open Access Journals (Sweden)

    Jian Xu

    Full Text Available BACKGROUND: The improvement of bone ingrowth into prosthesis and enhancement of the combination of the range between the bone and prosthesis are important for long-term stability of artificial joints. They are the focus of research on uncemented artificial joints. Porous materials can be of potential use to solve these problems. OBJECTIVES/PURPOSES: This research aims to observe the characteristics of the new porous Ti-25Nb alloy and its biocompatibility in vitro, and to provide basic experimental evidence for the development of new porous prostheses or bone implants for bone tissue regeneration. METHODS: The Ti-25Nb alloys with different porosities were fabricated using powder metallurgy. The alloys were then evaluated based on several characteristics, such as mechanical properties, purity, pore size, and porosity. To evaluate biocompatibility, the specimens were subjected to methylthiazol tetrazolium (MTT colorimetric assay, cell adhesion and proliferation assay using acridine staining, scanning electron microscopy, and detection of inflammation factor interleukin-6 (IL-6. RESULTS: The porous Ti-25Nb alloy with interconnected pores had a pore size of 200 µm to 500 µm, which was favorable for bone ingrowth. The compressive strength of the alloy was similar to that of cortical bone, while with the elastic modulus closer to cancellous bone. MTT assay showed that the alloy had no adverse reaction to rabbit bone marrow mesenchymal stem cells, with a toxicity level of 0 to 1. Cell adhesion and proliferation experiments showed excellent cell growth on the surface and inside the pores of the alloy. According to the IL-6 levels, the alloy did not cause any obvious inflammatory response. CONCLUSION: All porous Ti-25Nb alloys showed good biocompatibility regardless of the percentage of porosity. The basic requirement of clinical orthopedic implants was satisfied, which made the alloy a good prospect for biomedical application. The alloy with 70

  12. Flavonoids derived from Abelmoschus esculentus attenuatesUV-B Induced cell damage in human dermal fibroblasts throughNrf2-ARE pathway

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    Juilee Patwardhan

    2016-01-01

    Abbreviations used:ABTS: 2,2'-azino-bis-(3-ethylbenzothiazoline -6-sulphonic acid, AO: Acridine orange, ANOVA: Analysis of variance, ARE: Antioxidant response elements, BSA: Bovine serum albumin, CAPE: Caffeic acid phenethyl ester, CAT: Catalase, DCFH-DA: 2',7'-dichlorofluorescein diacetate, DMEM: Dulbecco's modified eagle's medium, DMSO: dimethyl sulfoxide, DNA: Deoxyribonucleic acid, DPBS: Dulbecco's phosphate-buffered saline, DPPH: 2,2-diphenyl-1-picryl hydrazyl, ECL: Enhanced chemiluminescence, EDTA: Ethylenediaminetetraacetic acid, ELISA: Enzyme-linked immunosorbent assay, EtBr: Ethidium bromide, FBS: Fetal bovine serum, FE Fraction: Flavonoid-enriched fraction, FRAP: Ferric reducing antioxidant power, GPx: Glutathione peroxidase, GR: Glutathione reductase, GST: Glutathione-S-transferase, GSH: Reduced glutathione, GSSG: Oxidized glutathione, HDF: Human dermal fibroblast adult cells, HEPES: 4-(2-hydroxyethyl-1-piperazineethanesulphonic acid, HRP: Horseradish peroxidase, HO-1: Heme oxygenase-1, HPTLC: High-performance thin layer chromatography, Keap-1: Kelch-like ECH-associated protein-1, MTT: 3-(4,5-dimethylthiazol-2-yl-2,5-diphenyltetrazolium bromide, NaCl: sodium chloride, NFDM: nonfat dry milk, Nrf2: Nuclear factor E2-related factor 2, NQO1: NAD (P H: Quinine oxidoreductase 1, OH: Hydroxyl ions, PBST: Phosphate-buffered saline with 0.1% tween 20, PCR: Polymerase chain reaction, PMSF: Phenylmethanesulfonyl fluoride, Rf: Retention factor, ROS: Reactive oxygen species, rRNA: Ribosomal ribonucleic acid, SDS: Sodium dodecyl sulfate, SOD: Superoxide dismutase, TLC: Thin layer chromatography, TLC-DPPH: Thin layer chromatography-2,2-diphenyl-1-picryl hydrazyl, UV: Ultraviolet, UV-A: Ultraviolet-A, UV-B: Ultraviolet-B, UV-C: Ultraviolet-C, qPCR: Quantitative polymerase chain reaction

  13. Characterization and applications of as-grown {beta}-Fe{sub 2}O{sub 3} nanoparticles prepared by hydrothermal method

    Energy Technology Data Exchange (ETDEWEB)

    Rahman, Mohammed M., E-mail: mmrahmanh@gmail.com; Jamal, Aslam; Khan, Sher Bahadar; Faisal, Mohd [Najran University, Department of Chemistry, Faculty of Sciences and Arts (Saudi Arabia)

    2011-09-15

    The production of low-dimensional nanoparticles (NPs) with appropriate surface modification has attracted increasing attention in biological, biochemical, and environmental applications including chemical sensing, photocatalytic degradation, separation, and purification of toxic molecules from the matrices. In this study, iron oxide NPs have been prepared by hydrothermal method using ferric chloride and urea in aqueous medium under alkaline condition (pH 9 {approx} 10). As-grown low-dimensional NPs have been characterized by UV-vis spectroscopy, FT-IR, X-ray diffraction, Field emission scanning electron microscopy, Raman spectroscopy, High-resolution Transmission electron microscopy, and Electron Diffraction System. The uniformity of the NPs size was measured by the scanning electron microscopy, while the single phase of the nanocrystalline {beta}-Fe{sub 2}O{sub 3} was characterized using powder X-ray diffraction technique. As-grown NPs were extensively applied for the photocatalytic degradation of acridine orange (AO) and electrochemical sensing of ammonia in liquid phase. Almost 50% photo-catalytic degradation with AO was observed in the presence of UV sources (250 W) with NPs. {beta}-Fe{sub 2}O{sub 3} NP-coated gold electrodes (GE, surface area 0.0216 cm{sup 2}) have enhanced ammonia-sensing performances in their electrical response (I-V characterization) for detecting ammonia in liquid phase. The performances of chemical sensor were investigated, and the results exhibited that the sensitivity, stability, and reproducibility of the sensor improved significantly using {beta}-Fe{sub 2}O{sub 3} NPs on GE surface. The sensitivity was approximately 0.5305 {+-} 0.02 {mu}Acm{sup -2}mM{sup -1}, with a detection limit of 21.8 {+-} 0.1 {mu}M, based on a signal/noise ratio of 3 with short response time.

  14. p21{sup WAF1/CIP1} deficiency induces mitochondrial dysfunction in HCT116 colon cancer cells

    Energy Technology Data Exchange (ETDEWEB)

    Kim, Ae Jeong; Jee, Hye Jin; Song, Naree; Kim, Minjee [Department of Biochemistry, College of Medicine, Dong-A University, Busan (Korea, Republic of); Mitochondria Hub Regulation Center, College of Medicine, Dong-A University, Busan (Korea, Republic of); Jeong, Seon-Young [Mitochondria Hub Regulation Center, College of Medicine, Dong-A University, Busan (Korea, Republic of); Department of Medical Genetics, Ajou University School of Medicine (Korea, Republic of); Yun, Jeanho, E-mail: yunj@dau.ac.kr [Department of Biochemistry, College of Medicine, Dong-A University, Busan (Korea, Republic of); Mitochondria Hub Regulation Center, College of Medicine, Dong-A University, Busan (Korea, Republic of)

    2013-01-11

    Highlights: Black-Right-Pointing-Pointer p21{sup -/-} HCT116 cells exhibited an increase in mitochondrial mass. Black-Right-Pointing-Pointer The expression levels of PGC-1{alpha} and AMPK were upregulated in p21{sup -/-} HCT116 cells. Black-Right-Pointing-Pointer The proliferation of p21{sup -/-} HCT116 cells in galactose medium was significantly impaired. Black-Right-Pointing-Pointer p21 may play a role in maintaining proper mitochondrial mass and respiratory function. -- Abstract: p21{sup WAF1/CIP1} is a critical regulator of cell cycle progression. However, the role of p21 in mitochondrial function remains poorly understood. In this study, we examined the effect of p21 deficiency on mitochondrial function in HCT116 human colon cancer cells. We found that there was a significant increase in the mitochondrial mass of p21{sup -/-} HCT116 cells, as measured by 10-N-nonyl-acridine orange staining, as well as an increase in the mitochondrial DNA content. In contrast, p53{sup -/-} cells had a mitochondrial mass comparable to that of wild-type HCT116 cells. In addition, the expression levels of the mitochondrial biogenesis regulators PGC-1{alpha} and TFAM and AMPK activity were also elevated in p21{sup -/-} cells, indicating that p21 deficiency induces the rate of mitochondrial biogenesis through the AMPK-PGC-1{alpha} axis. However, the increase in mitochondrial biogenesis in p21{sup -/-} cells did not accompany an increase in the cellular steady-state level of ATP. Furthermore, p21{sup -/-} cells exhibited significant proliferation impairment in galactose medium, suggesting that p21 deficiency induces a defect in the mitochondrial respiratory chain in HCT116 cells. Taken together, our results suggest that the loss of p21 results in an aberrant increase in the mitochondrial mass and in mitochondrial dysfunction in HCT116 cells, indicating that p21 is required to maintain proper mitochondrial mass and respiratory function.

  15. SMI of Bcl-2 TW-37 is active across a spectrum of B-cell tumors irrespective of their proliferative and differentiation status.

    Science.gov (United States)

    Al-Katib, Ayad M; Sun, Yuan; Goustin, Anton Scott; Azmi, Asfar Sohail; Chen, Ben; Aboukameel, Amro; Mohammad, Ramzi M

    2009-02-16

    The Bcl-2 family of proteins is critical to the life and death of malignant B-lymphocytes. Interfering with their activity using small-molecule inhibitors (SMI) is being explored as a new therapeutic strategy for treating B-cell tumors. We evaluated the efficacy of TW-37, a non-peptidic SMI of Bcl-2 against a range spectrum of human B-cell lines, fresh patient samples and animal xenograft models. Multiple cytochemical and molecular approaches such as acridine orange/ethidium bromide assay for apoptosis, co-immunoprecipitation of complexes and western blot analysis, caspase luminescent activity assay and apoptotic DNA fragmentation assay were used to demonstrate the effect of TW-37 on different B-cell lines, patient derived samples, as well as in animal xenograft models. Nanomolar concentrations of TW-37 were able to induce apoptosis in both fresh samples and established cell lines with IC50 in most cases of 165-320 nM. Apoptosis was independent of proliferative status or pathological classification of B-cell tumor. TW-37 was able to block Bim-Bcl-XL and Bim-Mcl-1 heterodimerization and induced apoptosis via activation of caspases -9, -3, PARP and DNA fragmentation. TW-37 administered to tumor-bearing SCID mice led to significant tumor growth inhibition (T/C), tumor growth delay (T-C) and Log10kill, when used at its maximum tolerated dose (40 mg/kg x 3 days) via tail vein. TW-37 failed to induce changes in the Bcl-2 proteins levels suggesting that assessment of baseline Bcl-2 family proteins can be used to predict response to the drug. These findings indicate activity of TW-37 across the spectrum of human B-cell tumors and support the concept of targeting the Bcl-2 system as a therapeutic strategy regardless of the stage of B-cell differentiation.

  16. Silencing of Bcl-XL Expression in Human MGC-803 Gastric Cancer Cells by siRNA

    Institute of Scientific and Technical Information of China (English)

    Xiao-Yong LEI; Miao ZHONG; Lan-Fang FENG; Chun-Yan YAN; Bing-Yang ZHU; Sheng-Song TANG; Duan-Fang LIAO

    2005-01-01

    To investigate the inhibitory effect of the Bcl-XL small interfering RNA (siRNA) on Bcl-XL gene expression in the human gastric cancer cell line MGC-803, green fluorescent protein (GFP) siRNA was constructed and transfected into MGC-803 ceils, together with GFP expression vector pTrace SV40.GFP expression levels were observed using fluorescence microscopy. Bcl-XL siRNA and negative siRNA were then constructed and stably transfected into MGC-803 cells. RT-PCR and immunofluorescence were used to detect the expression of Bcl-XL. Spontaneous apoptosis was detected by acridine orange (AO) and flow cytometry. Results were as follows: (1) 48 h after GFP expression vector and GFP siRNA co-transfection, the expression level of GFP in the GFP siRNA group was much lower than the negative siRNA group,according to fluorescence microscopy results. The mRNA and protein levels of Bcl-XL in Bcl-XL siRNA stable transfectants were reduced to almost background level compared with negative siRNA transfectants or untreated cells. (2) Changes in nucleus morphology was observed by AO staining nucleic and flow cytometry analysis, which showed that stable Bcl-XL siRNA transfectants have an increased spontaneous apoptosis (21.17%± 1.26% vs. 1.19%±0.18% and 1.56%±0.15 % respectively, P<0.05 vs. negative siRNA or untreated control). siRNA targeting GFP or Bcl-XL genes can specifically suppress GFP or Bcl-XL expression in MGC-803 cells, and Bcl-XL siRNA can increase spontaneous apoptosis. Bcl-XL siRNA may be a beneficial agent against human gastric adenocarcinoma.

  17. Orexin A induces autophagy in HCT-116 human colon cancer cells through the ERK signaling pathway.

    Science.gov (United States)

    Wen, Jing; Zhao, Yuyan; Guo, Lei

    2016-01-01

    Orexins are a class of peptides which have a potent influence on a broad variety of cancer cells. Autophagy is closely associated with tumors; however, its function is not yet completely understood. In this study, we aimed to determine whether orexin A induces autophagy in HCT‑116 human colon cancer cells and to elucidate the molecular mechanisms involved. For this purpose, HCT‑116 cells were treated with orexin A, and cell viability was then measured by MTT assay, and apoptosis was determined by flow cytometry. The expression levels of autophagy‑related proteins were measured by western blot analysis. Quantitative analysis of autophagy following acridine orange (AO) staining was performed using fluorescence microscopy, and cellular morphology was observed under a transmission electron microscope. In addition, the HCT‑116 cells were treated with the extracellular signal‑regulated kinase (ERK) inhibitor, U0126, or the autophagy inhibitor, chloroquine, in combination with orexin A in order to examine the activation of ERK. We found that orexin A significantly inhibited the viability of the HCT‑116 cells. Both autophagy and apoptosis were activated during the orexin A‑induced death of HCT‑116 cells. When the HCT‑116 cells were treated with orexin A for 24 h, an accumulation of punctate microtubule-associated protein-1 light chain 3 (LC3) and an increase in LC3‑Ⅱ protein levels were also detected, indicating the activation of autophagy. Moreover, orexin A upregulated ERK phosphorylation; however, U0126 or chloroquine abrogated ERK phosphorylation and decreased autophagy, compared to treatment with orexin A alone. Therefore, our findings demonstratedm that orexin A induced autophagy through the ERK pathway in HCT‑116 human colon cancer cells. The inhibition of autophagy may thus prove to be an effective strategy for enhancing the antitumor potential of orexin A as a treatment for colon cancer.

  18. Design real-time reversal of tumor multidrug resistance cleverly with shortened carbon nanotubes

    Directory of Open Access Journals (Sweden)

    Wu P

    2014-12-01

    Full Text Available Pingping Wu,1 Shang Li,2 Haijun Zhang2 1Jiangsu Cancer Hospital, Nanjing, People’s Republic of China; 2Department of Oncology, Zhongda Hospital, Medical School, Southeast University, Nanjing, People’s Republic of ChinaAbstract: Multidrug resistance (MDR in tumors renders many currently available chemotherapeutic drugs ineffective. Research in nanobiotechnology-based therapeutic alternatives has provided innovative and promising strategies to overcome MDR. The aim of this study was to investigate whether the new strategy of a co-loaded reversal agent and chemotherapeutic drug with shortened carbon nanotubes (CNTs would show useful effects on the real-time reversal of tumor MDR. CNTs were cut and purified via ultrasonication and oxidative acid treatment to optimize their length for drug-delivery vehicles, then verapamil (Ver and doxorubicin (Dox were co-loaded on shortened CNTs (denoted as Ver/Dox/shortened CNTs, which acted as a drug delivery system. The multidrug resistant leukemia K562/A02 cells were treated with the denoted Ver/Dox/shortened CNTs. The real-time reversal of tumor MDR were evaluated by flow cytometer, 3-(4,5-Dimethylthiazol-2-yl-2,5-diphenyltetrazolium bromide assays, acridine orange/ethidium bromide staining, and Western blot analysis. In the same MDR tumor cells the new strategy of a co-loaded reversal agent and chemotherapeutic drug with CNTs could inhibit the function of P-glycoprotein in real-time by Ver as reversal agent, significantly increase the uptake of Dox, enhance the sensitivity of the MDR cancer cells to the chemotherapeutic agent, and induce apoptosis. It was therefore concluded that a co-loaded reversal agent and chemotherapeutic drug with shortened CNTs could have real-time reversal ability of MDR in tumors, which could represent a promising approach in cancer therapy.Keywords: multidrug resistance, carbon nanotubes, drug delivery system, tumor

  19. Corosolic acid analogue, a natural triterpenoid saponin, induces apoptosis on human hepatocarcinoma cells through mitochondrial pathway in vitro.

    Science.gov (United States)

    Qu, Liping; Zhang, Huiqing; Yang, Yanlong; Yang, Geliang; Xin, Hailiang; Ling, Changquan

    2016-08-01

    Context 2a,-3a,-24-Trihydroxyurs-12-en-28-oic acid (TEO, a corosolic acid analogue) is a triterpenoid saponin isolated from Actinidia valvata Dunn (Actinidiaceae), a well-known traditional Chinese medicine. Objective This study investigated the anti-proliferation and inducing apoptosis effects of TEO in three human hepatocellular carcinoma (HCC) cell lines. Materials and methods Cytotoxic activity of TEO was determined by the MTT assay at various concentrations from 2.5 to 40 μg/mL in BEL-7402, BEL-7404 and SMMC-7721 cell lines. Cell morphology was assessed by acridine orange/ethidium bromide and 4'-6-diamidino-2-phenylindole dihydrochloride staining and fluorescence microscopy. Cell-cycle distribution and DNA damage were determined by flow cytometry and comet assay. Mitochondrial dysfunction was assessed by JC-1 staining and transmission electron microscopy. Apoptosis changes were explored by Western blot, TNF-α and caspase-3, -8, -9 assays. Results TEO exhibited inhibition effects on BEL-7402, BEL-7404 and SMMC-7721 cells treated for 24 h, the IC50 values were 34.6, 30.8 and 30.5 μg/mL, respectively. TEO (40 μg/mL)-treated three cell lines increased by more than 21% in the G1 phase and presented the morphological change and DNA damage. TEO also declined the mitochondrial membrane potential and altered mitochondrial ultra-structure. Furthermore, caspase-3, caspase-8, caspase-9 and TNF-α were also activated. Mechanism investigation showed that TEO could decrease anti-apoptotic Bcl-2 protein expression, increase proapoptotic Bax and Bid proteins expressions and increase Bax/Bcl-2 ratio. Conclusion Our results demonstrate for the first time that TEO inhibited growth of HCC cell lines and induced G1 phase arrest. Moreover, proapoptotic effects of TEO were mediated through the activation of TNF-α, caspases and mitochondrial pathway. PMID:26810384

  20. Assessment of the in vitro cytotoxicity and in vivo anti-tumor activity of the alcoholic stem bark extract/fractions of Mimusops elengi Linn.

    Science.gov (United States)

    Kumar, Harish; Savaliya, Mihir; Biswas, Subhankar; Nayak, Pawan G; Maliyakkal, Naseer; Manjunath Setty, M; Gourishetti, Karthik; Pai, K Sreedhara Ranganath

    2016-08-01

    Various parts of Mimusops elengi Linn. (Sapotaceae) have been used widely in traditional Indian medicine for the treatment of pain, inflammation and wounds. The study was conducted to explore the use of stem bark of M. elengi on pharmacological grounds and to evaluate the scientific basis of cytotoxic and anti-tumor activity. Extract/fractions were prepared and in vitro cytotoxicity was assessed using SRB assay. Most effective fractions were subjected to fluorescence microscopy based acridine orange/ethidium bromide (AO/EB) and Hoechst 33342 staining to determine apoptosis induction and DNA fragmentation assay. Comet and micronuclei assay were performed to assess genotoxicity. Cell cycle analysis was also performed. In vivo anti-tumor potential was evaluated by Ehrlich ascites carcinoma (EAC) model in mice. The alcoholic stem bark extract of M. elengi along with four fractions showed potential in vitro cytotoxicity in SRB assay. Of these, dichloromethane and ethyl acetate fractions were selected for further studies. The fractions revealed apoptosis inducing potential in AO/EB and Hoechst 33342 staining, which was further confirmed by DNA fragmentation assay. Genotoxic potential was revealed by comet and micronuclei assay. Fractions also exhibited specific cell cycle inhibition in G0/G1 phase. In EAC model, ethyl acetate fraction along with the standard (cisplatin) effectively reduced the increase in body weight compared to control and improved mean survival time. Both fractions were able to restore the altered hematological and biochemical parameters. Hence, M. elengi stem bark may be a possible therapeutic candidate having cytotoxic and anti-tumor potential. PMID:25701190

  1. Omeprazole inhibits proliferation and modulates autophagy in pancreatic cancer cells.

    Directory of Open Access Journals (Sweden)

    Andrej Udelnow

    Full Text Available BACKGROUND: Omeprazole has recently been described as a modulator of tumour chemoresistance, although its underlying molecular mechanisms remain controversial. Since pancreatic tumours are highly chemoresistant, a logical step would be to investigate the pharmacodynamic, morphological and biochemical effects of omeprazole on pancreatic cancer cell lines. METHODOLOGY/PRINCIPAL FINDINGS: Dose-effect curves of omeprazole, pantoprazole, gemcitabine, 5-fluorouracil and the combinations of omeprazole and 5-fluorouracil or gemcitabine were generated for the pancreatic cancer cell lines MiaPaCa-2, ASPC-1, Colo357, PancTu-1, Panc1 and Panc89. They revealed that omeprazole inhibited proliferation at probably non-toxic concentrations and reversed the hormesis phenomena of 5-fluorouracil. Electron microscopy showed that omeprazole led to accumulation of phagophores and early autophagosomes in ASPC-1 and MiaPaCa-2 cells. Signal changes indicating inhibited proliferation and programmed cell death were found by proton NMR spectroscopy of both cell lines when treated with omeprazole which was identified intracellularly. Omeprazole modulates the lysosomal transport pathway as shown by Western blot analysis of the expression of LAMP-1, Cathepsin-D and β-COP in lysosome- and Golgi complex containing cell fractions. Acridine orange staining revealed that the pump function of the vATPase was not specifically inhibited by omeprazole. Gene expression of the autophagy-related LC3 gene as well as of Bad, Mdr-1, Atg12 and the vATPase was analysed after treatment of cells with 5-fluorouracil and omeprazole and confirmed the above mentioned results. CONCLUSIONS: We hypothesise that omeprazole interacts with the regulatory functions of the vATPase without inhibiting its pump function. A modulation of the lysosomal transport pathway and autophagy is caused in pancreatic cancer cells leading to programmed cell death. This may circumvent common resistance mechanisms of

  2. HPLC detection of loss rate and cell migration of HUVECs in a proanthocyanidin cross-linked recombinant human collagen-peptide (RHC)–chitosan scaffold

    International Nuclear Information System (INIS)

    Porous scaffolds with appropriate pore structure, biocompatibility, mechanical property and processability play an important role in tissue engineering. In this paper, we fabricated a recombinant human collagen-peptide (RHC)–chitosan scaffold cross-linked by premixing 30% proanthocyanidin (PA) in one-step freeze-drying. To remove the residual acetic acid, optimized 0.2 M phosphate buffer of pH 6.24 with 30% ethanol (PBSE) was selected to neutralize the lyophilized scaffold followed by three times deionized water rinse. Ninhydrin assay was used to characterize the components loss during the fabrication process. To detect the exact RHC loss under optimized neutralization condition, high performance liquid chromatography (HPLC) equipped size exclusion chromatography column was used and the total RHC loss rate through PBSE rinse was 19.5 ± 5.08%. Fourier transform infrared spectroscopy (FT-IR) indicated hydrogen bonding among RHC, chitosan and PA, it also presented a probative but not strong hydrophobic interaction between phenyl rings of polyphenols and pyrrolidine rings of proline in RHC. Further, human umbilical vein endothelial cell (HUVEC) viability analyzed by a scanning electron microscope (SEM) and acridine orange/ethidium bromide (AO/EB) fluorescence staining exhibited that this scaffold could not only promote cell proliferation on scaffold surface but also permit cells migration into the scaffold. qRT-PCR exhibited that the optimized scaffold could stimulate angiogenesis associated genes VEGF and CD31 expression. These characterizations indicated that this scaffold can be considered as an ideal candidate for tissue engineering. - Highlights: • PA cross-linked recombinant human collagen–chitosan scaffold. • Fabrication in one-step lyophilization with neutralization. • HPLC detection of RHC loss rate • HUVEC proliferation and migration in scaffold • Angiogenesis associated gene expressions were increased in scaffold cell culturing

  3. Synthesis and anticancer activities of 4-(4-substituted piperazin)-5,6,7-trialkoxy quinazoline derivatives.

    Science.gov (United States)

    Zhang, Ying; Huang, Yin-Jiu; Xiang, Hong-Mei; Wang, Pei-Yi; Hu, De-Yu; Xue, Wei; Song, Bao-An; Yang, Song

    2014-05-01

    A series of 4-(4-substituted piperazin)-5,6,7-trialkoxy quinazoline was prepared by conventional heating methods. Among these compounds, the crystal structure of compound 10o (CCDC: 916922) was determined by X-ray crystallography. Bioassay results showed that most target compounds had certain inhibition activities against proliferation of tumor cells, and some compounds even had good broad-spectrum inhibition activities. The ethoxyl series of compounds possessed higher inhibition activities against tumor cells than the methoxyl series of compounds. Bioactivity tests showed that the IC50 values of compound 10s against PC3, MGC803, A375, and A549 cells were 1.8, 2.8, 1.3, and 2.9 μΜ, respectively, which were much higher than those of commercial gefitinib (7.2, 7.6, 7.2, and 9.8 μM, respectively). Conversely, the IC50 values of compound 10s were very low against NH3T3, indicating only weak effect on normal cells as also proven by lactate dehydrogenase and acridine orange/ethidium bromide staining. Analyses of cell configuration and cell cycle revealed that compound 10s possibly caused cells to remain at G0/G1 phase by inhibiting cell proliferation for 24 h. Compound 10s also inhibited the phosphorylation of ERK1/2 and P38 with obvious concentration dependence. Thus, these compounds can inhibit the proliferation of A549 cells through the interruption of ERK1/2 and P38signaling pathways.

  4. Laboratory diagnosis of tick-borne African relapsing fevers: latest developments

    Directory of Open Access Journals (Sweden)

    Aurélien eFotso Fotso

    2015-11-01

    Full Text Available In Africa, relapsing fevers caused by ectoparasite-borne Borrelia species are transmitted by ticks, with the exception of Borrelia recurrentis, which is a louse-borne spirochete. These tropical diseases responsible for mild to deadly spirochetemia. Cultured B. crocidurae, B. duttonii and B. hispanica circulate alongside at least six species which have not yet been cultured in vectors. Direct diagnosis is hindered by the use of non-specific laboratory tools. Indeed, microscopic observation of Borrelia spirochaeta in smears of peripheral blood taken from febrile patients lacks sensitivity and specificity. Although best visualised using dark-field microscopy, the organisms can also be detected using Wright-Giemsa or acridine orange stains.. PCR-based detection of specific sequences in total DNA extracted from a specimen can be used to discriminate different relapsing fever Borreliae. In our laboratory, we developed a multiplex real-time PCR assay for the specific detection of B. duttonii/recurrentis and B. crocidurae: Multispacer Sequence Typing accurately identified cultured relapsing fever borreliae and revealed diversity among them. Other molecular typing techniques, such as multilocus sequence analysis of tick-borne relapsing fever borreliae, showed the potential risk of human infection in Africa. Recent efforts to culture and sequence relapsing fever borreliae have provided new information for reassessment of the diversity of these bacteria. Recently, matrix-assisted laser desorption ionization time-of-flight mass spectrometry has been reported as a means of identifying cultured borreliae and of identifying both vectors and vectorised pathogens such as detecting relapsing fever borreliae directly in ticks. The lack of a rapid diagnosis test restricts the management of such diseases. We produced monoclonal antibodies against Borrelia crocidurae in order to develop cheap assays for the rapid detection of relapsing fever borreliae. In this paper

  5. Microwave-mediated extracellular synthesis of metallic silver and zinc oxide nanoparticles using macro-algae (Gracilaria edulis) extracts and its anticancer activity against human PC3 cell lines.

    Science.gov (United States)

    Priyadharshini, Ramaramesh Indra; Prasannaraj, Govindaraj; Geetha, Natesan; Venkatachalam, Perumal

    2014-12-01

    A rapid and novel microwave-mediated protocol was established for extracellular synthesis of metallic silver (Ag) and zinc oxide (ZnO) nanoparticles using the extracts of macro-algae Gracilaria edulis (GE) and also examined its anticancer activity against human prostate cancer cell lines (PC3). The formation of silver nanoparticles (GEAgNPs) and zinc oxide nanoparticles (GEZnONPs) in the reaction mixture was determined by ultraviolet-visible spectroscopy. The synthesized Ag and ZnO nanoparticles were characterized by X-ray diffraction, Fourier transform infra-red spectroscopy, energy dispersive X-ray, and field emission scanning electron microscopy. The silver and zinc oxide nanoparticles were spherical and rod-shaped, respectively. Cell viability assays were carried out to determine the cytotoxic effects of AgNPs and ZnONPs against PC3 and normal African monkey kidney (VERO) cell line. The inhibitory concentration values were found to be 39.60, 28.55, 53.99 μg/mL and 68.49, 88.05, 71.98 μg/mL against PC3 cells and Vero cells for AgNPs, ZnONPs, and aqueous G. edulis extracts, respectively, at 48 h incubation period. As evidenced by acridine orange/ethidium bromide staining, the percentage of the apoptotic bodies was found to be 62 and 70 % for AgNPs and ZnONPs, respectively. The present results strongly suggest that the synthesized ZnONPs showed an effective anticancer activity against PC3 cell lines than AgNPs.

  6. Bridged bis(beta-cyclodextrin)s possessing coordinated metal center(s) and their inclusion complexation behavior with model substrates: enhanced molecular binding ability by multiple recognition.

    Science.gov (United States)

    Liu, Y; Chen, Y; Li, L; Zhang, H Y; Liu, S X; Guan, X D

    2001-12-14

    To investigate quantitatively the cooperative binding ability of several beta-cyclodextrin oligomers bearing single or multiligated metal center(s), the inclusion complexation behavior of four bis(beta-cyclodextrin)s (2-5) linked by 2,2'-bipyridine-4,4'-dicarboxy tethers and their copper(II) complexes (6-9) with representative dye guests, i.e., methyl orange (MO), acridine red (AR), rhodamine B (RhB), ammonium 8-anilino-1-naphthalenesulfonic acid (ANS), and sodium 6-(p-toludino)-2-naphthalenesulfonate (TNS), have been examined in aqueous solution at 25 degrees C by means of UV-vis, circular dichroism, fluorescence, and 2D NMR spectroscopy. The results obtained indicate that bis(beta-cyclodextrin)s 2-5 can associate with one or three copper(II) ion(s) producing 2:1 or 2:3 bis(beta-cyclodextrin)-copper(II) complexes. These metal-ligated oligo(beta-cyclodextrin)s can bind two model substrates to form intramolecular 2:2 host-guest inclusion complexes and thus significantly enhance the original binding abilities of parent beta-cyclodextrin and bis(beta-cyclodextrin) toward model substrates through the cooperative binding of two guest molecules by four tethered cyclodextrin moieties, as well as the additional binding effect supplied by ligated metal center(s). Host 6 showed the highest enhancement of the stability constant, up to 38.3 times for ANS as compared with parent beta-cyclodextrin. The molecular binding mode and stability constant of substrates by bridged bis- and oligo(beta-cyclodextrin)s 2-9 are discussed from the viewpoint of the size/shape-fit interaction and molecular multiple recognition between host and guest.

  7. Effects of anticancer drugs on transcription in vitro.

    Science.gov (United States)

    Wilmańska, D; Czyz, M; Studzian, K; Piestrzeniewicz, M K; Gniazdowski, M

    2001-01-01

    The effects of DNA interacting drugs on: (1) total RNA synthesis catalyzed by E. coli and T7 RNA polymerase; (2) synthesis of the initiating dinucleotide (pppApU) by E. coli RNA polymerase ("abortive initiation"); (3) elongation of RNA chains synthesized by T7 RNA polymerase on pT7-7 plasmid DNA bearing T7 RNA polymerase promoter phi 10 with human Cu/Zn superoxide dismutase coding sequence, (4) interaction of transcription factor Sp1 and its binding site were studied. Intercalating ligands which form quickly dissociating complexes with DNA (anthracyclines, proflavine, ethidium bromide) are compared with the slowly dissociating drug of d(G x C) specificity (actinomycin D), the non-intercalating, d(A x T) specific pyrrole antibiotics (netropsin and distamycin A) and covalently binding to DNA 1-nitroacridine derivative (nitracrine). The obtained results indicate that rapidly dissociating ligands, proflavine and ethidium bromide, inhibit total RNA synthesis in vitro and the abortive initiation to a similar extent while they do not induce discrete elongation stops of RNA polymerase. Actinomycin D and nitracrine exhibit a high inhibitory effect on total RNA synthesis and induce stops of RNA polymerase while not affecting abortive initiation. Pyrrole antibiotics primarily inhibit the initiation, while no elongation stops are induced. Actinomycin D inhibits complex formation between nuclear proteins and the Sp1 binding site. Netropsin, ethidium bromide, proflavine and other intercalating acridines do not affect Sp1 binding. The results indicate that the effects primarily depend on sequence specificity and secondarily on the dissociation rate of ligands from their complexes with DNA. PMID:11724400

  8. Characterization of Crew Refuse Returned from Shuttle Missions with Permanent Gas, Volatile Organic Compound, and Microbial Analyses

    Science.gov (United States)

    Peterson, B.; Hummerick, M.; Roberts, M.; Krummins, V.; Kish, A.; Garland, J.; Maxwell, S.; Mills, A.

    In addition to the mass and energy costs associated with bioregenerative systems for advanced life support, the storage and processing of waste on spacecraft requires both atmospheric and biological management. Risks to crew health may arise from the presence of potential human pathogens in waste or from decay processes during waste storage and/or processing. This study reports on the permanent gas, trace volatile organic and microbiological analyses of crew refuse returned from shuttle missions STS-105, 109 and 110. The research objective is to characterize the biological stability of the waste stream, to assess the risks associated with its storage, and to provide baseline measures for the evaluation of waste processing technologies. Microbiological samples were collected from packaging material, food waste, bathroom waste, and bulk liquid collected from the volume F waste container. The number of culturable bacteria and total bacteria were determined by plating on R2A media and by Acridine Orange direct count, respectively. Samples of the trash were analyzed for the presence of fecal and total coliforms and other human-associated bacteria. Dry and ash weights were determined to estimate both water and organic content of the materials. The aerobic and anaerobic bio-stability of stored waste was determined by on-line monitoring of CO2 and by laboratory analysis of off-gas samples for hydrogen sulfide and methane. Volatile organic compounds and permanent gases were analyzed using EPA method TO15 with gas chromatography/mass spectrometry and by gas chromatography with selective detectors . This study establishes a baseline measure of waste composition, labile organics, and microbial load for this material.

  9. Susceptible cytotoxicity to ultraviolet B light in fibroblasts and keratinocytes cultured from autoimmune-prone MRL/Mp-lpr/lpr mice

    Energy Technology Data Exchange (ETDEWEB)

    Furukawa, F.; Lyon, M.B.; Norris, D.A. (Univ. of Colorado School of Medicine, Denver (USA))

    1989-09-01

    The MRL/Mp-lpr/lpr (MRL/l) mouse is an autoimmune model of spontaneous lupus erythematosus (LE), in addition to lupus nephritis. In order to better understand the mechanisms of photosensitivity in LE, in vitro photocytotoxicity was examined by using fibroblasts and keratinocytes cultured from MRL/l mice, control MRL/Mp- +/+ (MRL/n) mice, and normal BALB/c mice. A colorimetric 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide assay and the acridine orange/ethidium bromide assay were used for determination of cytotoxicity. Fibroblasts cultured from newborn MRL/l mice showed higher susceptibility to single ultraviolet light B (UVB) light irradiation at a dose of 100-500 mJ than those from MRL/n, F1 hybrid of (MRL/l x MRL/n mice), and BALB/c mice. However, the susceptibility to UVB was not observed in young (1-month-old) and adult (4-month-old) MRL/l mice. UVA light irradiation was not cytotoxic. Keratinocytes cultured from MRL mice showed lower cytotoxicity to UVB irradiation than fibroblasts cultured. However, keratinocytes from newborn MRL/l mice showed higher cytotoxicity to 50 mJ UVB irradiation than cells from MRL/n mice. Syngeneic or allogeneic sera augmented UVB-induced cytotoxicity of fibroblasts cultured. UVB irradiation of spleen cells induced no significant difference of cytotoxicity between MRL/l and MRL/n mice. Based on the results of F1 hybrid of (MRL/l x MRL/n) mice, the susceptibility seemed to be associated with autoimmune traits and to be regulated by genetical background.

  10. Platelet-rich plasma and fibrin glue-coated bioactive ceramics enhance growth and differentiation of goat bone marrow-derived stem cells.

    Science.gov (United States)

    Nair, Manitha B; Varma, H K; John, Annie

    2009-07-01

    New biotechnologies such as tissue engineering require functionally active cells within supportive matrices where the physical and chemical stimulus provided by the matrix is indispensable to determine the cellular behavior. This study has investigated the influence of platelet-rich plasma (PRP) and fibrin glue (FG) on the functional activity of goat bone marrow-derived mesenchymal stem cells (gBMSCs) that differentiated into the osteogenic lineage. To achieve this goal, PRP and FG were separately coated on bioactive ceramics like hydroxyapatite (HA) and silica-coated HA (HASi), on which gBMSCs were seeded and induced to differentiate into the osteogenic lineage for 28 days. The cells were then analyzed for viability (lactate dehydrogenase assay: acridine orange and ethidium bromide staining), morphology (scanning electron microscopy), proliferation (picogreen assay), cell cycle assay (propidium iodide staining), and differentiation (alkaline phosphatase [ALP] activity and real-time PCR analysis of ALP, osteocalcin, and osteopontin gene). It has been observed that PRP and FG have appreciably favored the viability, spreading, and proliferation of osteogenic-induced gBMSCs. The osteopontin and osteocalcin expression was significantly enhanced on PRP- and FG-coated HA and HASi, but PRP had effect on neither ALP expression nor ALP activity. The results of this study have depicted that FG-coated ceramics were better than PRP-coated and bare matrices. Among all, the excellent performance was shown by FG coated HASi, which may be attributed to the communal action of the stimulus emanated by Si in HASi and the temporary extracellular matrix provided by FG over HASi. Thus, we can conclude that PRP or FG in combination with bioactive ceramics could possibly enhance the functional activity of cells to a greater extent, promoting the hybrid composite as a promising candidate for bone tissue engineering applications.

  11. Fluorescent antibody application in bioremediation procedures at the Savannah River Site

    International Nuclear Information System (INIS)

    Direct Fluorescent Antibodies (DFA) and Most Probable Number (MPN) techniques are currently being employed at the Savannah River Site to monitor methanotrophic bacteria for the bioremediation of trichloroethylene (TCE) in field studies. Direct Fluorescent Antibodies were developed against various methanotrophic bacteria isolated from SRS as well as methanotrophic bacteria acquired from the American Type Culture Collection (ATCC). DFA's are anticipated to be more efficient for monitoring methanotroph activity than MPN's because of shorter processing time, lower cost, and the direct nature of the assay. The DFA method is a direct technique, in that samples are processed immediately and can be enumerated within an hour. The MPN method is indirect, since samples must be cultured for 6-8 weeks before measuring methane consumption and carbon dioxide production. Indirect methods are not highly selective and have limited application. The greatest advantage of a faster assay, is that bioremediation procedures utilizing methanotrophic bacteria could be amended. These amendments would be based on environmental monitoring with results in real time (1 hour). The elimination of the MPN technique and the use of DFA's will save significantly on both materials and labor. The data obtained from the DFA's and MPN's were statistically compared to each other and to total bacterial counts (AODC). The statistical analysis used was Analysis of Variants (ANOVA). Using this analysis, groundwater samples were found to be not significantly different; whereas soil were significantly different. These methods were employed on soil samples from the Southern Sector and ground water samples from the TCE-contaminated Sanitary Landfill at SRS. Acridine Orange Direct Counts were compared to show relative differences between total bacterial and methanotroph population

  12. Disulfiram attenuates osteoclast differentiation in vitro: a potential antiresorptive agent.

    Directory of Open Access Journals (Sweden)

    Hua Ying

    Full Text Available Disulfiram (DSF, a cysteine modifying compound, has long been clinically employed for the treatment of alcohol addiction. Mechanistically, DSF acts as a modulator of MAPK and NF-κB pathways signaling pathways. While these pathways are crucial for osteoclast (OC differentiation, the potential influence of DSF on OC formation and function has not been directly assessed. Here, we explore the pharmacological effects of DSF on OC differentiation, activity and the modulation of osteoclastogenic signaling cascades. We first analyzed cytotoxicity of DSF on bone marrow monocytes isolated from C57BL/6J mice. Upon the establishment of optimal dosage, we conducted osteoclastogenesis and bone resorption assays in the presence or absence of DSF treatment. Luciferase assays in RAW264.7 cells were used to examine the effects of DSF on major transcription factors activation. Western blot, reverse transcription polymerase chain reaction, intracellular acidification and proton influx assays were employed to further dissect the underlying mechanism. DSF treatment dose-dependently inhibited both mouse and human osteoclastogenesis, especially at early stages of differentiation. This inhibition correlated with a decrease in the expression of key osteoclastic marker genes including CtsK, TRAP, DC-STAMP and Atp6v0d2 as well as a reduction in bone resorption in vitro. Suppression of OC differentiation was found to be due, at least in part, to the blockade of several key receptor activators of nuclear factor kappa-B ligand (RANKL-signaling pathways including ERK, NF-κB and NFATc1. On the other hand, DSF failed to suppress intracellular acidification and proton influx in mouse and human osteoclasts using acridine orange quenching and microsome-based proton transport assays. Our findings indicate that DSF attenuates OC differentiation via the collective suppression of several key RANKL-mediated signaling cascades, thus making it an attractive agent for the treatment of OC

  13. Reliable Screening of Dye Phototoxicity by Using a Caenorhabditis elegans Fast Bioassay.

    Directory of Open Access Journals (Sweden)

    Javier Ignacio Bianchi

    Full Text Available Phototoxicity consists in the capability of certain innocuous molecules to become toxic when subjected to suitable illumination. In order to discover new photoactive drugs or characterize phototoxic pollutants, it would be advantageous to use simple biological tests of phototoxicy. In this work, we present a pilot screening of 37 dyes to test for phototoxic effects in the roundworm Caenorhabditis elegans. Populations of this nematode were treated with different dyes, and subsequently exposed to 30 min of white light. Behavioral outcomes were quantified by recording the global motility using an infrared tracking device (WMicrotracker. Of the tested compounds, 17 dyes were classified as photoactive, being phloxine B, primuline, eosin Y, acridine orange and rose Bengal the most phototoxic. To assess photoactivity after uptake, compounds were retested after washing them out of the medium before light irradiation. Dye uptake into the worms was also analyzed by staining or fluorescence. All the positive drugs were incorporated by animals and produced phototoxic effects after washing. We also tested the stress response being triggered by the treatments through reporter strains. Endoplasmic reticulum stress response (hsp-4::GFP strain was activated by 22% of phototoxic dyes, and mitochondrial stress response (hsp-6::GFP strain was induced by 16% of phototoxic dyes. These results point to a phototoxic perturbation of the protein functionality and an oxidative stress similar to that reported in cell cultures. Our work shows for the first time the feasibility of C. elegans for running phototoxic screenings and underscores its application on photoactive drugs and environmental pollutants assessment.

  14. Activation of the intrinsic-apoptotic pathway in LNCaP prostate cancer cells by genistein- topotecan combination treatments

    Directory of Open Access Journals (Sweden)

    Vanessa Hörmann

    2013-03-01

    Full Text Available ABSTRACTBackground: Prostate cancer is the second most common cancer in American men. The development of alternative preventative and/or treatment options utilizing a combination of phytochemicals and chemotherapeutic drugs could be an attractive alternative compared to conventional carcinoma treatments. Genistein isoflavone is the primary dietary phytochemical found in soy and has demonstrated anti-tumor activities in LNCaP prostate cancer cells. Topotecan Hydrochloride (Hycamtin is an FDA-approved chemotherapy for secondary treatment of lung, ovarian and cervical cancers. The purpose of this study was to detail the potential activation of the intrinsic apoptotic pathway in LNCaP prostate cancer cells through genistein-topotecan combination treatments.Methods: LNCaP cells were cultured in complete RPMI medium in a monolayer (70-80% confluency at 37ºC and 5% CO2. Treatment consisted of single and combination groups of genistein and topotecan for 24 hours. The treated cells were assayed for i growth inhibition through trypan blue exclusion assay and microphotography , ii classification of cellular death through acridine/ ethidium bromide fluorescent staining, and iii activation of the intrinsic apoptotic pathway through Jc-1: mitochondrial membrane potential assay, cytochrome c release and Bcl-2 protein expression.Results: The overall data indicated that genistein-topotecan combination was significantly more efficacious in reducing the prostate carcinoma’s viability compared to the single treatment options. In all treatment groups, cell death occurred primarily through the activation of the intrinsic apoptotic pathway.Conclusion: The combination of topotecan and genistein has the potential to lead to treatment options with equal therapeutic efficiency as traditional chemo- and radiation therapies, but lower cell cytotoxicity and fewer side effects in patients.

  15. Autophagy as a Survival Mechanism for Squamous Cell Carcinoma Cells in Endonuclease G-Mediated Apoptosis

    Science.gov (United States)

    Masui, Atsushi; Hamada, Masakazu; Kameyama, Hiroyasu; Wakabayashi, Ken; Takasu, Ayako; Imai, Tomoaki; Iwai, Soichi; Yura, Yoshiaki

    2016-01-01

    Safingol, L- threo-dihydrosphingosine, induces cell death in human oral squamous cell carcinoma (SCC) cells through an endonuclease G (endoG) -mediated pathway. We herein determined whether safingol induced apoptosis and autophagy in oral SCC cells. Safingol induced apoptotic cell death in oral SCC cells in a dose-dependent manner. In safingol-treated cells, microtubule-associated protein 1 light chain 3 (LC3)-I was changed to LC3-II and the cytoplasmic expression of LC3, amount of acidic vesicular organelles (AVOs) stained by acridine orange and autophagic vacuoles were increased, indicating the occurrence of autophagy. An inhibitor of autophagy, 3-methyladenine (3-MA), enhanced the suppressive effects of safingol on cell viability, and this was accompanied by an increase in the number of apoptotic cells and extent of nuclear fragmentation. The nuclear translocation of endoG was minimal at a low concentration of safingol, but markedly increased when combined with 3-MA. The suppressive effects of safingol and 3-MA on cell viability were reduced in endoG siRNA- transfected cells. The scavenging of reactive oxygen species (ROS) prevented cell death induced by the combinational treatment, whereas a pretreatment with a pan-caspase inhibitor z-VAD-fmk did not. These results indicated that safingol induced apoptosis and autophagy in SCC cells and that the suppression of autophagy by 3-MA enhanced apoptosis. Autophagy supports cell survival, but not cell death in the SCC cell system in which apoptosis occurs in an endoG-mediated manner. PMID:27658240

  16. Green synthesis of silver and gold nanoparticles from Gymnema sylvestre leaf extract: study of antioxidant and anticancer activities

    Energy Technology Data Exchange (ETDEWEB)

    Nakkala, Jayachandra Reddy; Mata, Rani; Bhagat, Ekta; Sadras, Sudha Rani, E-mail: dr.ssrlab@gmail.com, E-mail: sadrassudha@gmail.com [Pondicherry University, Department of Biochemistry and Molecular Biology, School of Life Sciences (India)

    2015-03-15

    The present study reports the biological synthesis of silver and gold nanoparticles from Gymnema sylvestre leaf extract and their in vitro free radical scavenging efficacy as well as antiproliferative effect in Hep2 cells. The formation of silver (GYAgNPs) and gold nanoparticles (GYAuNPs) was confirmed by UV–visible spectroscopy. The average size of synthesized GYAgNPs and GYAuNPs was found to be 33 and 26 nm, respectively, by DLS particle size analyzer. TEM analysis indicated spherical shape of GYAgNPs and GYAuNPs and in EDX analysis they produced strong signal for silver and gold, respectively. Both GYAgNPs and GYAuNPs exhibited strong in vitro free radical quenching ability and their activity was comparable to that of GYLE. The cytotoxic effect of GYAgNPs and GYAuNPs in Hep2 cells was examined by MTT assay in which GYAgNPs displayed an IC{sub 50} value of 121 µg ml{sup −1}, while GYAuNPs produced up to 38 % of inhibition at the maximum concentration of 250 µg ml{sup −1} used in this study. Distinct morphological changes were observed in Hep2 cells following treatment with GYAgNPs and GYAuNPs at 24 h, and orange-colored apoptotic bodies were located by acridine orange and ethidium bromide double-staining technique. Also, there was increase in the levels of reactive oxygen species in treated cells as indicated by 2′,7′-dichlorofluorescin diacetate staining. Further, nuclear changes like chromatin condensation/fragmentation were also observed by propidium iodide and 4′,6-diamidino-2-phenylindole dilactate staining methods. These findings support that the antiproliferative effects of GYAgNPs and GYAuNPs in Hep2 cells are mediated through induction of apoptosis.

  17. Inhibition of mTOR-dependent autophagy sensitizes leukemic cells to cytarabine-induced apoptotic death.

    Directory of Open Access Journals (Sweden)

    Mihajlo Bosnjak

    Full Text Available The present study investigated the role of autophagy, a cellular self-digestion process, in the cytotoxicity of antileukemic drug cytarabine towards human leukemic cell lines (REH, HL-60, MOLT-4 and peripheral blood mononuclear cells from leukemic patients. The induction of autophagy was confirmed by acridine orange staining of intracellular acidic vesicles, electron microscopy visualization of autophagic vacuoles, as well as by the increase in autophagic proteolysis and autophagic flux, demonstrated by immunoblot analysis of p62 downregulation and LC3-I conversion to autophagosome-associated LC3-II in the presence of proteolysis inhibitors, respectively. Moreover, the expression of autophagy-related genes Atg4, Atg5 and Atg7 was stimulated by cytarabine in REH cells. Cytarabine reduced the phosphorylation of the major negative regulator of autophagy, mammalian target of rapamycin (mTOR, and its downstream target p70S6 kinase in REH cells, which was associated with downregulation of mTOR activator Akt and activation of extracellular signal- regulated kinase. Cytarabine had no effect on the activation of mTOR inhibitor AMP-activated protein kinase. Leucine, an mTOR activator, reduced both cytarabine-induced autophagy and cytotoxicity. Accordingly, pharmacological downregulation of autophagy with bafilomycin A1 and chloroquine, or RNA interference-mediated knockdown of LC3β or p62, markedly increased oxidative stress, mitochondrial depolarization, caspase activation and subsequent DNA fragmentation and apoptotic death in cytarabine-treated REH cells. Cytarabine also induced mTOR-dependent cytoprotective autophagy in HL-60 and MOLT-4 leukemic cell lines, as well as primary leukemic cells, but not normal leukocytes. These data suggest that the therapeutic efficiency of cytarabine in leukemic patients could be increased by the inhibition of the mTOR-dependent autophagic response.

  18. Lunar and Martian soil stimulants have different effects on L-[14C]glutamate binding to brain nerve terminals

    Science.gov (United States)

    Borisova, Tatiana; Krisanova, Natalia; Nazarova, Anastasiya; Borysov, Arseniy; Chunihin, Olexander

    Nano-sized particles can be deleterious to human physiology because they may be internalized by lung epithelium and overcome the blood-brain barrier. The health effects from exposure to Lunar and Martian dust are almost completely unknown, whereas they can be deleterious to human physiology. The effects of Lunar and Martian Soil Simulants (Orbital Technologies Corporation, Madison, USA) on the conductance of planar lipid membrane, membrane potential, acidification of synaptic vesicles, glutamate uptake, and ambient level of glutamate in isolated rat brain nerve terminals (synaptosomes) were studied using photon correlation spectroscopy, Planar Lipid Bilayer technique, spectrofluorimetry, radiolabeled assay, respectively. Lunar and Martian Soil Simulants did not influence the conductance of planar lipid membrane. It was revealed that nerve terminals were not indifferent to the exposure to inorganic particles of Lunar and Martian Soil Simulants. Using Zetasizer Nanosystem (Malvern Instruments) with helium-neon laser for dynamic light scattering (DLS), the synaptosomal size before and after the addition of Lunar and Martian Soil Simulants was measured and the binding of Lunar and Martian Soil Simulants inorganic particles to nerve terminals was demonstrated. Using potential-sensitive fluorescent dye rhodamine 6G, we showed that Lunar and Martian Soil particles did not influence the potential of the plasma membrane of nerve terminals. Acidification of synaptic vesicles of nerve terminals was not changed in the presence of Lunar and Martian Soil particles that was revealed with pH-sensitive fluorescent dye acridine orange. Martian Soil Simulant particles did not change binding of L-[14C]glutamate to brain nerve terminals, in contrast, Lunar ones changed this parameter and this fact may have harmful consequences to human physiology, in particular, glutamate homeostasis in the mammalian CNS.

  19. 1-(2,6-Dihydroxy-4-methoxyphenyl-2-(4-hydroxyphenyl Ethanone-Induced Cell Cycle Arrest in G1/G0 in HT-29 Cells Human Colon Adenocarcinoma Cells

    Directory of Open Access Journals (Sweden)

    Ma Ma Lay

    2014-01-01

    Full Text Available 1-(2,6-Dihydroxy-4-methoxyphenyl-2-(4-hydroxyphenyl ethanone (DMHE was isolated from the ethyl acetate fraction of Phaleria macrocarpa (Scheff. Boerl fruits and the structure confirmed by GC-MS (gas chromatography-mass spectrometry and NMR (nuclear magnetic resonance analysis. This compound was tested on the HT-29 human colon adenocarcinoma cell line using MTT (method of transcriptional and translational cell proliferation assay. The results of MTT assay showed that DMHE exhibited good cytotoxic effect on HT-29 cells in a dose- and time-dependent manner but no cytotoxic effect on the MRC-5 cell line after 72 h incubation. Morphological features of apoptotic cells upon treatment by DMHE, e.g., cell shrinkage and membrane blebbing, were examined by an inverted and phase microscope. Other features, such as chromatin condension and nuclear fragmentation were studied using acridine orange and propidium iodide staining under the fluorescence microscope. Future evidence of apoptosis/necrosis was provided by result fromannexin V-FITC/PI (fluorescein-isothiocyanate/propidium iodide staining revealed the percentage of early apoptotic, late apoptotic, necrotic and live cells in a dose- and time-dependent manner using flow cytometry. Cell cycle analysis showed G0/G1 arrest in a time-dependent manner. A western blot analysis indicated that cell death might be associated with the up-regulation of the pro-apoptotic proteins Bax PUMA. However, the anit-apotptic proteins Bcl-2, Bcl-xL, and Mcl-1 were also found to increase in a time-dependent manner. The expression of the pro-apoptotic protein Bak was not observed.

  20. Nestin is essential for zebrafish brain and eye development through control of progenitor cell apoptosis.

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    Hua-Ling Chen

    Full Text Available BACKGROUND: Nestin is expressed in neural progenitor cells (NPC of developing brain. Despite its wide use as an NPC marker, the function of nestin in embryo development is unclear. METHODOLOGY/PRINCIPAL FINDINGS: As nestin is conserved in zebrafish and its predicted sequence is clustered with the mammalian nestin orthologue, we used zebrafish as a model to investigate its role in embryogenesis. Injection of nestin morpholino (MO into fertilized eggs induced time- and dose-dependent brain and eye developmental defects. Nestin morphants exhibited characteristic morphological changes including small head, small eyes and hydrocephalus. Histological examinations show reduced hind- and mid-brain size, dilated ventricle, poorly organized retina and underdeveloped lens. Injection of control nestin MO did not induce brain or eye changes. Nestin MO injection reduced expression of ascl1b (achaete-scute complex-like 1b, a marker of NPCs, without affecting its distribution. Nestin MO did not influence Elavl3/4 (Embryonic lethal, abnormal vision, Drosophila-like 3/4 (a neuronal marker, or otx2 (a midbrain neuronal marker, but severely perturbed cranial motor nerve development and axon distribution. To determine whether the developmental defects are due to excessive NPC apoptosis and/or reduced NPC proliferation, we analyzed apoptosis by TUNEL assay and acridine orange staining and proliferation by BrdU incorporation, pcna and mcm5 expressions. Excessive apoptosis was noted in hindbrain and midbrain cells. Apoptotic signals were colocalized with ascl1b. Proliferation markers were not significantly altered by nestin MO. CONCLUSION/SIGNIFICANCE: These results suggest that nestin is essential for zebrafish brain and eye development probably through control of progenitor cell apoptosis.

  1. Penetration of erythromycin through Staphylococcus epidermidis biofilm

    Institute of Scientific and Technical Information of China (English)

    LIN Mao-hu; HE Lei; GAO Jie; LIU Yun-xi; SUO Ji-jiang; XING Yu-bin; JIA Ning

    2013-01-01

    Background The catheter related infection caused by Staphylococcus epiderrnidis biofilm is increasing and difficult to treat by antimicrobial chemotherapy.The properties of biofilms that give rise to antibiotic resistance are only partially understood.This study aimed to elucidate the penetration of erythromycin through Staphylococcus epidermidis biofilm.Methods The penetration ratio of erythromycin through Staphylococcus epidermidis biofilms of 1457,1457-msrA,and wild isolate S68 was detected by biofilm penetration model at different time points according to the standard regression curve.The RNNDNA ratio and the cell density within the biofilms were observed by confocal laser microscope and transmission electromicroscope,respectively.Results The penetration ratios of erythromycin through the biofilms of 1457,1457-msrA,and S68 after cultivation for 36 hours were 0.93,0.55 and 0.4,respectively.The erythromycin penetration ratio through 1457 biofilm (0.58 after 8 hours)was higher than that through the other two (0.499 and 0.31 after 24 hours).Lower growth rate of the cells in biofilm was shown,with reduction of RNA/DNA proportion observed by confocal laser microscope through acridine orange stain.Compared with the control group observed by transmission electrmicroscope,the cell density of biofilm air face was lower than that of agar face,with more cell debris.Conclusions Erythromycin could penetrate to the Staphylococcus epidermidis biofilm,but could not kill the cells thoroughly.The lower growth rate of the cells within biofilm could help decreasing the erythromycin susceptibility.

  2. A preliminary experimental study on the cardiac toxicity of glutamate and the role of α-amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid receptor in rats

    Institute of Scientific and Technical Information of China (English)

    LIU Yan; ZHOU Lan; XU Hai-fei; YAN Li; DING Fan; HAO Wei; CAO Ji-min

    2013-01-01

    Background Monosodium L-glutamate (MSG) is a food flavour enhancer and its potential harmfulness to the heart remains controversial.We investigated whether MSG could induce cardiac arrhythmias and apoptosis via the α-amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid (AMPA) receptor.Methods Myocardial infarction (MI) was created by ligating the coronary artery and ventricular arrhythmias were monitored by electrocardiogram in the rat in vivo.Neonatal rat cardiomyocytes were isolated and cultured.Cell viability was estimated by 3-(4,5)-dimethylthiahiazo(-z-yl)-3,5-di-phenytetrazoliumromide (MTT) assay.Calcium mobilization was monitored by confocal microscopy.Cardiomyocyte apoptosis was evaluated by acridine orange staining,flow cytometry,DNA laddering,reverse transcription-polymerase chain reaction (RT-PCR) and Western blotting.Results MSG (i.v.) decreased the heart rate at 0.5 g/kg and serious bradycardia at 1.5 g/kg,but could not induce ventricular tachyarrhythmias in normal rats in vivo.In rats with acute MI in vivo,however,MSG (1.5 g/kg,i.v.) induced ventricular tachyarrhythmias and these arrhythmias could be prevented by blocking the AMPA and N-methyl-d-aspartate (NMDA) receptors.Selectively activating the AMPA or NMDA receptor induced ventricular tachyarrhythmias in MI rats.At the cellular level,AMPA induced calcium mobilization,oxidative stress,mitochondrial dysfunction and apoptosis in cultured cardiomyocytes,especially when the AMPA receptor desensitization were blocked by cyclothiazide.The above toxic cellular effects of AMPA were abolished by AMPA receptor blockade or by H2O2 scavengers.Conclusions MSG induces bradycardia in normal rats,but triggers lethal tachyarrhythmias in myocardial infarcted rats probably by hindering AMPA receptors.AMPA receptor overstimulation also induces cardiomyocyte apoptosis,which may facilitate arrhythmia.

  3. Matrine induces the hepatic differentiation of WB-F344 rat hepatic progenitor cells and inhibits Jagged 1/HES1 signaling.

    Science.gov (United States)

    Yang, Zhiyun; Wang, Li; Wang, Xianbo

    2016-10-01

    Matrine is a Chinese medicine, which is widely utilized for the attenuation of liver injuries and promotion of liver regeneration. It was previously observed that the in vivo administration of matrine promoted oval cell‑mediated liver regeneration in a rat model, suggesting that this compound may affect the differentiation of hepatic progenitor cells. The present study aimed to determine the mechanisms underlying this observation and to investigate the effect of matrine on the differentiation of the WB‑F344 rat hepatic progenitor cell line. Matrine was administered to rats, and rat serum was collected. WB‑F344 cells were cultured in the presence or absence of the rat serum for 24‑72 h, and the effects on cell viability and proliferation were assessed using acridine orange/propidium iodide staining and a 3‑(4,5‑dimethylthiazol‑2‑yl) ‑2,5‑diphenyltetrazolium bromide assay. The expression of albumin (ALB, a hepatocyte marker) and the notch signaling pathway ligand, Jagged 1, were assessed using immunohistochemistry and western blotting, and the mRNA transcription of ALB, Jagged 1 and hairy and enhancer of split‑1 (HES1, another notch signaling ligand) were measured using reverse transcription‑polymerase chain reaction analysis. The results showed that proliferation of the WB‑F344 cells was inhibited by matrine serum in a concentration‑ and time‑dependent manner. Matrine serum downregulated Jagged 1 and HES1, and upregulated ALB, indicating the induction of WB‑F344 cell differentiation. The effects of matrine serum were reversed by supplementing the culture medium with 0.1 mol/l parathyroid hormone, a Notch signaling pathway activator. In conclusion, matrine induced hepatic differentiation of the hepatic progenitor cells, likely by inhibiting the Jagged 1/HES1 signaling pathway.

  4. An NMR study of sodium poly(acrylate) adsorption on rutile

    International Nuclear Information System (INIS)

    Adsorption of sodium poly(acrylate) (PA) on rutile particles in aqueous dispersion was studied. Two different molecular weights of PA (2,100 and 30,000) and two different grades of rutile were used. Various pHs, ionic strengths, and PA concentrations were investigated. The main technique employed was measurement of the transverse NMR relaxation of the solvent using the CPMG pulse sequence. Other techniques used to augment these results include electroacoustics, scanning electron microscopy, and measurement of the adsorbed amount of PA by a fluorescence spectroscopy technique using the dye acridine orange. Adsorption of small ions such as Na+, K+, Cl-, and NO3- to the particle surface was found to have a significant effect on the measured transverse relaxation rate, that was dependent on the pH and the concentration of the ions. There was usually an additional effect on the relaxation due to the adsorbed PA, but only qualitative rather than quantitative information about the adsorption could be deduced. At pH 4 especially, it could be seen that the results were consistent with the common assertion that polymers adsorb in a flat conformation at low concentration, and only become looped when all of the surface sites are full. At pH 10 it was found that the relaxation rate for the longer chain PA samples fluctuated over time, indicating metastable PA conformations. There were also unusual trends in the relaxation rate for these samples, which could be due to a previously proposed small ion complexation mechanism for PA adsorption at high pH in this system. It is possible that an extensive and comprehensive study using this technique, investigating all of the relevant parameters, especially the effect of small ion adsorption, may allow a quantitative description of the adsorbed conformation. (author)

  5. Effect of Sodium Arsenite on Rat Bone Marrow Mesenchymal Stem Cells: Cells Viability and Morphological Study

    Directory of Open Access Journals (Sweden)

    M.H. Abnosi

    2010-07-01

    Full Text Available Introduction & Objective: Sodium arsenite as an environmental pollutant being found in the air, water, and earth crust threats the human beings' health. The aim of this study was to investigate the effect of sodium arsenite on viability and morphology of mesenchymal stem cells in rat bone marrow.Materials & Methods: In this exprimental study the cells were extracted in DMEM containing 15% FBS and Pen/Strep until the 3rd passage then treated with 0, 0.1, 0.5, 2.5, 12.5 and 20 µM of sodium arsenite for 12, 24, 36 and 48 hrs. Viability of the cells was carried out with trypan blue and MTT staining, then 0.1 µM and 36 hrs treatment was selected for further investigations. Morphology of the cells was studied using fluorescent dye (Hochest, propidium iodide and acridine orange as well as protein profile of the cells were studied using SDS-PAGE. Data was analyzed using one and two way ANOVA.Results: Based on the two way ANOVA, cumulative effect of treatment time and used dosage caused highly significant reduction (p<0.001 in viability of rat bone marrow mesenchymal stem cells. One way ANOVA indicated that the viability of the cells reduced significantly (p<0.05 from 0.1 µM of sodium arsenite on wards in all the treatment time. Morphological changes including condensation and deformation of the nuclei, membrane disruption, and shrinkage of cytoplasm were also observed. Conclusion: Sodium arsenite toxicity caused morphological and protein profile changes as well as dose and time dependent reduction in viability of rat bone marrow mesenchymal stem cells.

  6. Combined effects of depleted uranium and ionising radiation on zebrafish embryos.

    Science.gov (United States)

    Ng, C Y P; Pereira, S; Cheng, S H; Adam-Guillermin, C; Garnier-Laplace, J; Yu, K N

    2015-11-01

    In the environment, living organisms are exposed to a mixture of stressors, and the combined effects are deemed as multiple stressor effects. In the present work, the authors studied the multiple stressor effect in embryos of the zebrafish (Danio rerio) from simultaneous exposure to alpha particles and depleted uranium (DU) through quantification of apoptotic signals at 24 h post-fertilisation (hpf) revealed by vital dye acridine orange staining. In each set of experiments, dechorionated zebrafish embryos were divided into 4 groups, each having 10 embryos: Group (C) in which the embryos did not receive any further treatment; Group (IU) in which the embryos received an alpha-particle dose of 0.44 mGy at 5 hpf and were then exposed to 100 µg l(-1) of DU from 5 to 6 hpf; Group (I) in which the embryos received an alpha-particle dose of 0.44 mGy at 5 hpf and Group (U) in which the dechorionated embryos were exposed to 100 µg l(-1) of DU from 5 to 6 hpf. The authors confirmed that an alpha-particle dose of 0.44 mGy and a DU exposure for 1 h separately led to hormetic and toxic effects assessed by counting apoptotic signals, respectively, in the zebrafish. Interestingly, the combined exposure led to an effect more toxic than that caused by the DU exposure alone, so effectively DU changed the beneficial effect (hormesis) brought about by alpha-particle irradiation into an apparently toxic effect. This could be explained in terms of the promotion of early death of cells predisposed to spontaneous transformation by the small alpha-particle dose (i.e. hormetic effect) and the postponement of cell death upon DU exposure. PMID:25948823

  7. BIO-ORGANIC CHEMISTRY QUARTERLY REPORT. June through August1963

    Energy Technology Data Exchange (ETDEWEB)

    Various

    1963-10-02

    This report covers the following titles: (1) The Effects of 8-Methyl Lipoic Acid on the Evolution of Oxygen and Reduction of Carbon Dioxide during Photosynthesis; (2) Further {sup 14}C and {sup 15}N Tracer Studies of Amino Acid Synthesis during Photosynthesis by Chlorella Pyrenoidosa; (3) Two-Dimensional High Voltage, Low-Temperature Paper Electrophoresis of {sup 14}C-Labeled Products of Photosynthesis with {sup 14}CO{sub 2}; (4) A Search for Enzymic and Nonenzymic Reactions Between Thiamine Derivatives and Sugar Phosphates; (5) The Cytochrome Content of Purified Spinach Chloroplast Lamellae; (6) The Osmium Tetroxide Fixation of Chloroplast Lamellae; (7) Kinetics of Exoenzymes and Applications to the Determination of the Sequence of Nucleic Acids; (8) Brain Biochemistry and Behavior in Rats; (9) Experiments on Classical Conditioning and Light Habituation in Planarians; (10) Operant Conditioning in Planarians; (11) Manganese Porphyrin Complexes; (12) EPR Studies of Some Complex Organic Solutions; (13) Transient Response of Light-induced Photosynthetic Electron Paramagnetic Resonance Signals: Rhodospirillum rubrum Chromatophores; (14) Studies of the Tautomerism of Amides; (15) Structure and Mechanism of Hydrolysis of the Product of Reaction of PZ05 and Ethyl Ether; (16) A Study of the Irradiation Products of Several Nitrones; (17) Biosynthesis of the Opium Alkaloids; (18) Synthesis of methyl-{beta}-D-thiogalactoside-{sup 35}S; (19) Effect of Acridine Orange and Visible Light on Thymine Dimer Formation and Disruption; (20) Some Aspects of the Radiation Chemistry of DNA; (21) Nuclear Magnetic Resonance; and (22) Studies on the Inhibition of the Photoreduction of FMN.

  8. Nucleolus-like bodies of fully-grown mouse oocytes contain key nucleolar proteins but are impoverished for rRNA.

    Science.gov (United States)

    Shishova, Kseniya V; Lavrentyeva, Elena A; Dobrucki, Jurek W; Zatsepina, Olga V

    2015-01-15

    It is well known that fully-grown mammalian oocytes, rather than typical nucleoli, contain prominent but structurally homogenous bodies called "nucleolus-like bodies" (NLBs). NLBs accumulate a vast amount of material, but their biochemical composition and functions remain uncertain. To clarify the composition of the NLB material in mouse GV oocytes, we devised an assay to detect internal oocyte proteins with fluorescein-5-isothiocyanate (FITC) and applied the fluorescent RNA-binding dye acridine orange to examine whether NLBs contain RNA. Our results unequivocally show that, similarly to typical nucleoli, proteins and RNA are major constituents of transcriptionally active (or non-surrounded) NLBs as well as of transcriptionally silent (or surrounded) NLBs. We also show, by exposing fixed oocytes to a mild proteinase K treatment, that the NLB mass in oocytes of both types contains nucleolar proteins that are involved in all major steps of ribosome biogenesis, including rDNA transcription (UBF), early rRNA processing (fibrillarin), and late rRNA processing (NPM1/nucleophosmin/B23, nucleolin/C23), but none of the nuclear proteins tested, including SC35, NOBOX, topoisomerase II beta, HP1α, and H3. The ribosomal RPL26 protein was detected within the NLBs of NSN-type oocytes but is virtually absent from NLBs of SN-type oocytes. Taking into account that the major class of nucleolar RNA is ribosomal RNA (rRNA), we applied fluorescence in situ hybridization with oligonucleotide probes targeting 18S and 28S rRNAs. The results show that, in contrast to active nucleoli, NLBs of fully-grown oocytes are impoverished for the rRNAs, which is consistent with the absence of transcribed ribosomal genes in the NLB mass. Overall, the results of this study suggest that NLBs of fully-grown mammalian oocytes serve for storing major nucleolar proteins but not rRNA.

  9. Antimicrobial nisin acts against saliva derived multi-species biofilms without cytotoxicity to human oral cells

    Science.gov (United States)

    Shin, Jae M.; Ateia, Islam; Paulus, Jefrey R.; Liu, Hongrui; Fenno, J. Christopher; Rickard, Alexander H.; Kapila, Yvonne L.

    2015-01-01

    Objectives: Nisin is a lantibiotic widely used for the preservation of food and beverages. Recently, investigators have reported that nisin may have clinical applications for treating bacterial infections. The aim of this study was to investigate the effects of ultra pure food grade Nisin ZP (>95% purity) on taxonomically diverse bacteria common to the human oral cavity and saliva derived multi-species oral biofilms, and to discern the toxicity of nisin against human cells relevant to the oral cavity. Methods: The minimum inhibitory concentrations and minimum bactericidal concentrations of taxonomically distinct oral bacteria were determined using agar and broth dilution methods. To assess the effects of nisin on biofilms, two model systems were utilized: a static and a controlled flow microfluidic system. Biofilms were inoculated with pooled human saliva and fed filter-sterilized saliva for 20–22 h at 37°C. Nisin effects on cellular apoptosis and proliferation were evaluated using acridine orange/ethidium bromide fluorescent nuclear staining and lactate dehydrogenase activity assays. Results: Nisin inhibited planktonic growth of oral bacteria at low concentrations (2.5–50 μg/ml). Nisin also retarded development of multi-species biofilms at concentrations ≥1 μg/ml. Specifically, under biofilm model conditions, nisin interfered with biofilm development and reduced biofilm biomass and thickness in a dose-dependent manner. The treatment of pre-formed biofilms with nisin resulted in dose- and time-dependent disruption of the biofilm architecture along with decreased bacterial viability. Human cells relevant to the oral cavity were unaffected by the treatment of nisin at anti-biofilm concentrations and showed no signs of apoptotic changes unless treated with much higher concentrations (>200 μg/ml). Conclusion: This work highlights the potential therapeutic value of high purity food grade nisin to inhibit the growth of oral bacteria and the development of

  10. Singlet oxygen mediated apoptosis by anthrone involving lysosomes and mitochondria at ambient UV exposure

    Energy Technology Data Exchange (ETDEWEB)

    Mujtaba, Syed Faiz [Photobiology Division, (CSIR)-Indian Institute of Toxicology Research, Post Box No. 80, M.G. Marg, Lucknow 226001, Uttar Pradesh (India); College of Pharmacy, Faculty of Pharmaceutical Sciences, Pt. B.D.S University of Health Sciences, Rohtak, Haryana (India); Dwivedi, Ashish; Yadav, Neera [Photobiology Division, (CSIR)-Indian Institute of Toxicology Research, Post Box No. 80, M.G. Marg, Lucknow 226001, Uttar Pradesh (India); Ray, R.S., E-mail: ratanray.2011@rediffmail.com [Photobiology Division, (CSIR)-Indian Institute of Toxicology Research, Post Box No. 80, M.G. Marg, Lucknow 226001, Uttar Pradesh (India); Singh, Gajendra [College of Pharmacy, Faculty of Pharmaceutical Sciences, Pt. B.D.S University of Health Sciences, Rohtak, Haryana (India)

    2013-05-15

    Highlights: ► Photomodification of anthrone at ambient environmental intensities of UV-radiation. ► Phototoxicity of anthrone through type-II photodynamic reaction by generating {sup 1}O{sub 2}. ► Role of DNA damage and lipid peroxidation in anthrone phototoxicity. ► Apototic cell death and involvement of lysosomes and mitochondria. ► Up-regulation of p21 and bax concomitantly down regulation of bcl2 genes expression. -- Abstract: Anthrone a tricyclic aromatic hydrocarbon which is toxic environmental pollutant comes in the environment through photooxidation of anthracene. We have studied the photomodification of anthrone under environmental conditions. Anthrone generates reactive oxygen species (ROS) like {sup 1}O{sub 2} through Type-II photodynamic reaction. Significant intracellular ROS generation was measured through dichlorohydrofluorescein fluorescence intensity. The generation of {sup 1}O{sub 2} was further substantiated by using specific quencher like sodium azide. UV induced photodegradation of 2-deoxyguanosine and photoperoxidation of linoleic acid accorded the involvement of {sup 1}O{sub 2} in the manifestation of anthrone phototoxicity. Phototoxicity of anthrone was done on human keratinocytes (HaCaT) through 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide and neutral red uptake assays. Anthrone induced cell cycle arrest (G2/M-phase) and DNA damage in a concentration dependent manner. We found apoptosis as a pattern of cell death which was confirmed through sub-G1 fraction, morphological changes, caspase-3 activation, acridine orange/ethidium bromide staining and phosphatidylserine translocation. Mitochondrial depolarization and lysosomal destabilization was parallel to apoptotic process. Our RT-PCR results strongly supports our view point of apoptotic cell death through up-regulation of pro-apoptotic genes p21 and Bax, and down regulation of anti-apoptotic gene Bcl{sub 2}. Therefore, much attention should be paid to concomitant

  11. Studies of ruthenium(II) polypyridyl complexes on cytotoxicity in vitro, apoptosis, DNA-binding and antioxidant activity

    Science.gov (United States)

    Huang, Hong-Liang; Liu, Yun-Jun; Zeng, Cheng-Hui; Yao, Jun-Hua; Liang, Zhen-Hua; Li, Zheng-Zheng; Wu, Fu-Hai

    2010-03-01

    Two new ruthenium(II) polypyridyl complexes [Ru(dmb) 2(maip)](ClO 4) 21 (maip = 2-(3-aminophenyl)imizado[4,5-f][1,10]phenanthroline and [Ru(dmb) 2(maip)](ClO 4) 22 (paip = 2-(4-aminophenyl)imidazo[4,5-f][1,10]phenanthroline, dmb = 4,4'-dimethyl-2,2'-bipyridine) have been synthesized and characterized. The DNA-binding behaviors of complexes 1 and 2 were studied by viscosity measurements, thermal denaturation, photocleavage, absorption titration and luminescence spectra. The results show that the two complexes intercalate between the base pairs of DNA. The DNA-binding constants Kb for complexes 1 and 2 were determined to be 1.12 ± 0.11 × 10 5 M -1 ( s = 2.17) and 3.46 ± 0.59 × 10 5 M -1 ( s = 2.11) M -1. The studies on the mechanism of photocleavage demonstrate that superoxide anion radical (O 2rad - ) and singlet oxygen ( 1O 2) may play an important role. The cytotoxicity of these complexes has been evaluated by MTT (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide) assay. The IC 50 values are 19.21, 33.15, 38.57 and 21.15 for complex 1 and 41.77, 123.58, 255.44 and 49.11 for complex 2 against BEL-7402, C-6, HepG-2 and MCF-7 cell lines, respectively. The apoptosis assay was carried out with acridine orange/ethidium bromide (AO/EB) staining methods and the results indicate that complexes can induce the apoptosis of BEL-7402 cells. The experiments on antioxidant activity show these complexes exhibit good antioxidant activity against hydroxyl radical (OH rad ).

  12. Singlet oxygen mediated apoptosis by anthrone involving lysosomes and mitochondria at ambient UV exposure

    International Nuclear Information System (INIS)

    Highlights: ► Photomodification of anthrone at ambient environmental intensities of UV-radiation. ► Phototoxicity of anthrone through type-II photodynamic reaction by generating 1O2. ► Role of DNA damage and lipid peroxidation in anthrone phototoxicity. ► Apototic cell death and involvement of lysosomes and mitochondria. ► Up-regulation of p21 and bax concomitantly down regulation of bcl2 genes expression. -- Abstract: Anthrone a tricyclic aromatic hydrocarbon which is toxic environmental pollutant comes in the environment through photooxidation of anthracene. We have studied the photomodification of anthrone under environmental conditions. Anthrone generates reactive oxygen species (ROS) like 1O2 through Type-II photodynamic reaction. Significant intracellular ROS generation was measured through dichlorohydrofluorescein fluorescence intensity. The generation of 1O2 was further substantiated by using specific quencher like sodium azide. UV induced photodegradation of 2-deoxyguanosine and photoperoxidation of linoleic acid accorded the involvement of 1O2 in the manifestation of anthrone phototoxicity. Phototoxicity of anthrone was done on human keratinocytes (HaCaT) through 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide and neutral red uptake assays. Anthrone induced cell cycle arrest (G2/M-phase) and DNA damage in a concentration dependent manner. We found apoptosis as a pattern of cell death which was confirmed through sub-G1 fraction, morphological changes, caspase-3 activation, acridine orange/ethidium bromide staining and phosphatidylserine translocation. Mitochondrial depolarization and lysosomal destabilization was parallel to apoptotic process. Our RT-PCR results strongly supports our view point of apoptotic cell death through up-regulation of pro-apoptotic genes p21 and Bax, and down regulation of anti-apoptotic gene Bcl2. Therefore, much attention should be paid to concomitant exposure of anthrone and UV-R for its total

  13. A vacuolar-type proton pump in a vesicle fraction enriched with potassium transporting plasma membranes from tobacco hornworm midgut

    Energy Technology Data Exchange (ETDEWEB)

    Wieczorek, H.; Weerth, S.; Schindlbeck, M.; Klein, U.

    1989-07-05

    Mg-ATP dependent electrogenic proton transport, monitored with fluorescent acridine orange, 9-aminoacridine, and oxonol V, was investigated in a fraction enriched with potassium transporting goblet cell apical membranes of Manduca sexta larval midgut. Proton transport and the ATPase activity from the goblet cell apical membrane exhibited similar substrate specificity and inhibitor sensitivity. ATP and GTP were far better substrates than UTP, CTP, ADP, and AMP. Azide and vanadate did not inhibit proton transport, whereas 100 microM N,N'-dicyclohexylcarbodiimide and 30 microM N-ethylmaleimide were inhibitors. The pH gradient generated by ATP and limiting its hydrolysis was 2-3 pH units. Unlike the ATPase activity, proton transport was not stimulated by KCl. In the presence of 20 mM KCl, a proton gradient could not be developed or was dissipated. Monovalent cations counteracted the proton gradient in an order of efficacy like that for stimulation of the membrane-bound ATPase activity: K+ = Rb+ much greater than Li+ greater than Na+ greater than choline (chloride salts). Like proton transport, the generation of an ATP dependent and azide- and vanadate-insensitive membrane potential (vesicle interior positive) was prevented largely by 100 microM N,N'-dicyclohexylcarbodiimide and 30 microM N-ethylmaleimide. Unlike proton transport, the membrane potential was not affected by 20 mM KCl. In the presence of 150 mM choline chloride, the generation of a membrane potential was suppressed, whereas the pH gradient increased 40%, indicating an anion conductance in the vesicle membrane. Altogether, the results led to the following new hypothesis of electrogenic potassium transport in the lepidopteran midgut. A vacuolar-type electrogenic ATPase pumps protons across the apical membrane of the goblet cell, thus energizing electroneutral proton/potassium antiport. The result is a net active and electrogenic potassium flux.

  14. The blockade of cyclooxygenases-1 and -2 reduces the effects of hypoxia on endothelial cells

    Directory of Open Access Journals (Sweden)

    Gloria M.A.

    2006-01-01

    Full Text Available Hypoxia activates endothelial cells by the action of reactive oxygen species generated in part by cyclooxygenases (COX production enhancing leukocyte transmigration. We investigated the effect of specific COX inhibition on the function of endothelial cells exposed to hypoxia. Mouse immortalized endothelial cells were subjected to 30 min of oxygen deprivation by gas exchange. Acridine orange/ethidium bromide dyes and lactate dehydrogenase activity were used to monitor cell viability. The mRNA of COX-1 and -2 was amplified and semi-quantified before and after hypoxia in cells treated or not with indomethacin, a non-selective COX inhibitor. Expression of RANTES (regulated upon activation, normal T cell expressed and secreted protein and the protective role of heme oxygenase-1 (HO-1 were also investigated by PCR. Gas exchange decreased partial oxygen pressure (PaO2 by 45.12 ± 5.85% (from 162 ± 10 to 73 ± 7.4 mmHg. Thirty minutes of hypoxia decreased cell viability and enhanced lactate dehydrogenase levels compared to control (73.1 ± 2.7 vs 91.2 ± 0.9%, P < 0.02; 35.96 ± 11.64 vs 22.19 ± 9.65%, P = 0.002, respectively. COX-2 and HO-1 mRNA were up-regulated after hypoxia. Indomethacin (300 µM decreased COX-2, HO-1, hypoxia-inducible factor-1alpha and RANTES mRNA and increased cell viability after hypoxia. We conclude that blockade of COX up-regulation can ameliorate endothelial injury, resulting in reduced production of chemokines.

  15. Cytotoxic and pro-apoptotic activities of leaf extract of Croton bonplandianus Baill. against lung cancer cell line A549.

    Science.gov (United States)

    Bhavana, J; Kalaivani, M K; Sumathy, A

    2016-06-01

    The acetone extract (AcE) of the Croton bonplandianus Baill., an exotic weed of the Euphorbiaceae family was studied for cytotoxicity, apoptosis, cell cycle arrest in A549 cell line and antioxidant capacities using MTT assay, acridine orange/ethidium bromide (AO/EB staining), cell cycle analysis and DPPH radical scavenging assay respectively. Based on the cytotoxic activity, the extract was tested for the apoptotic effect using AO/EB and Hoechst 33258 staining. The apoptosis was characterized by chromatin condensation and DNA fragmentation. Further, to determine the stage of cell death, cell cycle analysis was performed by flow cytometry and AcE was found to arrest G2/M phase in a dose dependent manner. The number of cells in G2/M phase increases with concurrent accumulation of cells in sub G₀/G₁phase indicates the induction of apoptosis at G2M phase. The free radical scavenging activity of the AcE against DPPH was considerably significant. The cytotoxic, apoptotic and antioxidant effect of the AcE could be well correlated with the presence of potent free radical scavenging secondary metabolites such as phenols (43 ± 0.05 µg/mL), flavonoids (3.5 ± 0.07 µg/mL) and tannin (0.36 ± 0.1 µg/mL). Our study has shown that A549 cells were more sensitive to AcE with an IC₅₀ of 15.68 ± 0.006 µg/mL compared to the standard drug 2.20 ± 0.008 µg/mL (cisplatin). The results suggest that Croton bonplandianus could serve as a potential source of alternative therapeutic agent for treating cancer. Further research is required to isolate the active principle compound and determination of its anticancer property.

  16. Reactive oxygen and nitrogen (ROS and RNS) species generation and cell death in tomato suspension cultures--Botrytis cinerea interaction.

    Science.gov (United States)

    Pietrowska, E; Różalska, S; Kaźmierczak, A; Nawrocka, J; Małolepsza, U

    2015-01-01

    This article reports events connected to cell survival and Botrytis cinerea infection development in cell suspension cultures of two tomato cultivars which show different levels of susceptibility to the pathogen: cv. Corindo (more susceptible) and cv. Perkoz (less susceptible). In parallel changes in reactive oxygen (ROS) and nitrogen (RNS) species generation and in S-nitrosoglutathione reductase (GSNOR) activity were studied. In vivo staining methods with acridine orange (AO) and ethidium bromide (EB) as well as fluorescent microscopy were used to assess tomato and B. cinerea cells death. The biochemical studies of ROS and RNS concentrations in plant cell extract were complemented by in vivo ROS and nitric oxide (NO) imaging using nitro blue tetrazolium (NBT), diaminobenzidine (DAB) and diaminofluorescein diacetate (DAF-DA) staining methods, and confocal microscope technique. B. cinerea infection proceeded slower in Perkoz cell cultures. It was evidenced by measuring the pathogen conidia germination and germination tube development in which nuclei revealing cell death dominated. Two different types of tomato cell death were observed: cells with necrotic nuclei dominated in Corindo whereas in Perkoz cells with characteristic of vacuolar death type prevailed. In Perkoz cells, constitutive levels of NO and S-nitrosothiols (SNO) were significantly higher and hydrogen peroxide (H₂O₂) and superoxide anion (O₂(-)) concentrations were slightly higher as compared with Corindo cells. Moreover, increases in these molecule concentrations as a result of B. cinerea inoculation were observed in both, Perkoz and Corindo cell cultures. The enzymatic GSNOR activity seems to be an important player in controlling the SNO level in tomato cells. Involvements of the studied compounds in molecular mechanisms of tomato resistance to B. cinerea are discussed in the paper. PMID:25064634

  17. The effects of PAHs and N-PAHs on retinoid signaling and Oct-4 expression in vitro.

    Science.gov (United States)

    Beníšek, Martin; Kubincová, Petra; Bláha, Luděk; Hilscherová, Klára

    2011-02-01

    Polycyclic aromatic hydrocarbons (PAHs) and their N-heterocyclic analogs (N-PAHs) are important environmental contaminants with negative effects in living organisms, including teratogenicity and embryotoxicity. Though most studies linked their embryotoxicity with aryl hydrocarbon receptor (AhR) and cytochrome P450 activation, the exact mechanism is not known. Other mechanisms such as disruption of retinoid signaling were recently suggested to be of importance. This study investigated PAHs and N-PAHs interference with retinoid signaling in vitro by modulating all-trans retinoic acid (ATRA) mediated response in a reporter gene assay using P19/A15 cell line. Further, effects on pluripotency and differentiation processes were evaluated by measuring octamer-4 (Oct-4), an important pluripotency marker and master differentiation factor. Two of the studied compounds, benz[a]anthracene and benz[c]acridine significantly up-regulated ATRA-mediated response in the co-exposure with a range of ATRA concentrations. Another structural N-PAH variant, 1,7-phenanthroline, downregulated ATRA-mediated response at most of tested ATRA concentrations and exposure times. Interesting concentration-dependent biphasic effects (i.e. downregulation with subsequent up-regulation to control levels) were observed at co-exposures of ATRA and parent PAH phenanthrene. Non significant Oct-4 modulation in co-exposure with ATRA was observed at compounds, which potentiated ATRA-mediated effects in the reporter gene assay. On the other hand, 1,7-phenanthroline and phenanthrene significantly suppressed Oct-4 levels in higher tested concentrations. Our results further extend the knowledge of PAH and N-PAH in vitro effects and indicate that these environmental toxicants may have influence on differentiation process and embryonic development by interfering with ATRA signaling and by modulating levels of Oct-4. PMID:21111795

  18. Homology modeling, molecular dynamics, and virtual screening of NorA efflux pump inhibitors of Staphylococcus aureus

    Science.gov (United States)

    Bhaskar, Baki Vijaya; Babu, Tirumalasetty Muni Chandra; Reddy, Netala Vasudeva; Rajendra, Wudayagiri

    2016-01-01

    Emerging drug resistance in clinical isolates of Staphylococcus aureus might be implicated to the overexpression of NorA efflux pump which is capable of extruding numerous structurally diverse compounds. However, NorA efflux pump is considered as a potential drug target for the development of efflux pump inhibitors. In the present study, NorA model was constructed based on the crystal structure of glycerol-3-phosphate transporter (PDBID: 1PW4). Molecular dynamics (MD) simulation was performed using NAMD2.7 for NorA which is embedded in the hydrated lipid bilayer. Structural design of NorA unveils amino (N)- and carboxyl (C)-terminal domains which are connected by long cytoplasmic loop. N and C domains are composed of six transmembrane α-helices (TM) which exhibits pseudo-twofold symmetry and possess voluminous substrate binding cavity between TM helices. Molecular docking of reserpine, totarol, ferruginol, salvin, thioxanthene, phenothiazine, omeprazole, verapamil, nalidixic acid, ciprofloxacin, levofloxacin, and acridine to NorA found that all the molecules were bound at the large hydrophobic cleft and indicated significant interactions with the key residues. In addition, structure-based virtual screening was employed which indicates that 14 potent novel lead molecules such as CID58685302, CID58685367, CID5799283, CID5578487, CID60028372, ZINC12196383, ZINC72140751, ZINC72137843, ZINC39227983, ZINC43742707, ZINC12196375, ZINC66166948, ZINC39228014, and ZINC14616160 have highest binding affinity for NorA. These lead molecules displayed considerable pharmacological properties as evidenced by Lipinski rule of five and prophecy of toxicity risk assessment. Thus, the present study will be helpful in designing and synthesis of a novel class of NorA efflux pump inhibitors that restore the susceptibilities of drug compounds. PMID:27757014

  19. The role of apoptosis in MCLR-induced developmental toxicity in zebrafish embryos

    Energy Technology Data Exchange (ETDEWEB)

    Zeng, Cheng [College of Fisheries, Huazhong Agricultural University, Wuhan 430070 (China); Sun, Hong [Hubei Maternal and Child Health Hospital, Wuhan 430070 (China); Xie, Ping [Donghu Experimental Station of Lake Ecosystems, State Key Laboratory for Freshwater Ecology and Biotechnology of China, Institute of Hydrobiology, The Chinese Academy of Sciences, Wuhan 430072 (China); Wang, Jianghua; Zhang, Guirong; Chen, Nan [College of Fisheries, Huazhong Agricultural University, Wuhan 430070 (China); Yan, Wei, E-mail: Yanwei75126@163.com [Institute of Agricultural Quality Standards and Testing Technology, Hubei Academy of Agricultural Sciences, Wuhan 430064 (China); Li, Guangyu, E-mail: ligy2001@163.com [College of Fisheries, Huazhong Agricultural University, Wuhan 430070 (China); Freshwater Aquaculture Collaborative Innovation Center of Hubei Province, Wuhan 430070 (China)

    2014-04-01

    Highlights: • MCLR-induced apoptosis in the heart of developing embryos leads to the growth delay in zebrafish. • MCLR-triggered apoptosis might be induced by ROS. • P53–Bax–Bcl-2 and caspase-dependent apoptotic pathway contribute greatly to MCLR-induced apoptosis. Abstract: We previously demonstrated that cyanobacteria-derived microcystin–leucine–arginine (MCLR) is able to induce developing toxicity, such as malformation, growth delay and also decreased heart rates in zebrafish embryos. However, the molecular mechanisms by which MCLR induces its toxicity during the development of zebrafish remain largely unknown. Here, we evaluate the role of apoptosis in MCLR-induced developmental toxicity. Zebrafish embryos were exposed to various concentrations of MCLR (0, 0.2, 0.5, 2, and 5.0 mg L⁻¹ for 96 h, at which time reactive oxygen species (ROS) was significantly induced in the 2 and 5.0 mg L⁻¹ MCLR exposure groups. Acridine orange (AO) staining and terminal deoxynucleotide transferase-mediated deoxy-UTP nick end labelling (TUNEL) assay showed that MCLR exposure resulted in cell apoptosis. To test the apoptotic pathway, the expression pattern of several apoptotic-related genes was examined for the level of enzyme activity, gene and protein expression, respectively. The overall results demonstrate that MCLR induced ROS which consequently triggered apoptosis in the heart of developing zebrafish embryos. Our results also indicate that the p53–Bax–Bcl-2 pathway and the caspase-dependent apoptotic pathway play major roles in MCLR-induced apoptosis in the developing embryos.

  20. Structure of the AcrAB-TolC multidrug efflux pump.

    Science.gov (United States)

    Du, Dijun; Wang, Zhao; James, Nathan R; Voss, Jarrod E; Klimont, Ewa; Ohene-Agyei, Thelma; Venter, Henrietta; Chiu, Wah; Luisi, Ben F

    2014-05-22

    The capacity of numerous bacterial species to tolerate antibiotics and other toxic compounds arises in part from the activity of energy-dependent transporters. In Gram-negative bacteria, many of these transporters form multicomponent 'pumps' that span both inner and outer membranes and are driven energetically by a primary or secondary transporter component. A model system for such a pump is the acridine resistance complex of Escherichia coli. This pump assembly comprises the outer-membrane channel TolC, the secondary transporter AcrB located in the inner membrane, and the periplasmic AcrA, which bridges these two integral membrane proteins. The AcrAB-TolC efflux pump is able to transport vectorially a diverse array of compounds with little chemical similarity, thus conferring resistance to a broad spectrum of antibiotics. Homologous complexes are found in many Gram-negative species, including in animal and plant pathogens. Crystal structures are available for the individual components of the pump and have provided insights into substrate recognition, energy coupling and the transduction of conformational changes associated with the transport process. However, how the subunits are organized in the pump, their stoichiometry and the details of their interactions are not known. Here we present the pseudo-atomic structure of a complete multidrug efflux pump in complex with a modulatory protein partner from E. coli. The model defines the quaternary organization of the pump, identifies key domain interactions, and suggests a cooperative process for channel assembly and opening. These findings illuminate the basis for drug resistance in numerous pathogenic bacterial species. PMID:24747401

  1. Streptococcus oralis Induces Lysosomal Impairment of Macrophages via Bacterial Hydrogen Peroxide.

    Science.gov (United States)

    Okahashi, Nobuo; Nakata, Masanobu; Kuwata, Hirotaka; Kawabata, Shigetada

    2016-07-01

    Streptococcus oralis, an oral commensal, belongs to the mitis group of streptococci and occasionally causes opportunistic infections, such as bacterial endocarditis and bacteremia. Recently, we found that the hydrogen peroxide (H2O2) produced by S. oralis is sufficient to kill human monocytes and epithelial cells, implying that streptococcal H2O2 is a cytotoxin. In the present study, we investigated whether streptococcal H2O2 impacts lysosomes, organelles of the intracellular digestive system, in relation to cell death. S. oralis infection induced the death of RAW 264 macrophages in an H2O2-dependent manner, which was exemplified by the fact that exogenous H2O2 also induced cell death. Infection with either a mutant lacking spxB, which encodes pyruvate oxidase responsible for H2O2 production, or Streptococcus mutans, which does not produce H2O2, showed less cytotoxicity. Visualization of lysosomes with LysoTracker revealed lysosome deacidification after infection with S. oralis or exposure to H2O2, which was corroborated by acridine orange staining. Similarly, fluorescent labeling of lysosome-associated membrane protein-1 gradually disappeared during infection with S. oralis or exposure to H2O2 The deacidification and the following induction of cell death were inhibited by chelating iron in lysosomes. Moreover, fluorescent staining of cathepsin B indicated lysosomal destruction. However, treatment of infected cells with a specific inhibitor of cathepsin B had negligible effects on cell death; instead, it suppressed the detachment of dead cells from the culture plates. These results suggest that streptococcal H2O2 induces cell death with lysosomal destruction and then the released lysosomal cathepsins contribute to the detachment of the dead cells. PMID:27113357

  2. Artonin E Induces Apoptosis via Mitochondrial Dysregulation in SKOV-3 Ovarian Cancer Cells

    Science.gov (United States)

    Karimian, Hamed; Dehghan, Firouzeh; Nordin, Noraziah; Mohd Ali, Hapipah; Mohan, Syam; Mohd Hashim, Najihah

    2016-01-01

    Artonin E is a prenylated flavonoid isolated from the stem bark of Artocarpus elasticus Reinw.(Moraceae). This study aimed to investigate the apoptotic mechanisms induced by artonin E in a metastatic human ovarian cancer cell line SKOV-3 in vitro. MTT assay, clonogenic assay, acridine orange and propidium iodide double staining, cell cycle and annexin V analyses were performed to explore the mode of artonin E-induced cell death at different time points. DNA laddering, activation of caspases-3, -8, and -9, multi-parametric cytotoxicity-3analysis by high-content screening, measurement of reactive oxygen species generation, and Western blot were employed to study the pathways involved in the apoptosis. MTT results showed that artonin E inhibited the growth of SKOV-3 cells, with IC50 values of 6.5±0.5μg/mL after 72 h treatment, and showed less toxicity toward a normal human ovarian cell lineT1074, with IC50 value of 32.5±0.5μg/mL. Results showed that artonin E induced apoptosis and cell cycle arrest at the S phase. This compound also promoted the activation of caspases-3, -8, and -9. Further investigation into the depletion of mitochondrial membrane potential and release of cytochrome c revealed that artonin E treatment induced apoptosis via regulation of the expression of pro-survival and pro-apoptotic Bcl-2 family members. The expression levels of survivin and HSP70 proteins were also down regulated in SKOV-3 cells treated with artonin E. We propose that artonin E induced an antiproliferative effect that led to S phase cell cycle arrest and apoptosis through dysregulation of mitochondrial pathways, particularly the pro- and anti-apoptosis signaling pathways. PMID:27019365

  3. Binding of 8-methoxypsoralen to DNA in vitro: Monitoring by spectroscopic and chemometrics approaches

    Energy Technology Data Exchange (ETDEWEB)

    Zhou, Xiaoyue; Zhang, Guowen, E-mail: gwzhang@ncu.edu.cn; Wang, Langhong

    2014-10-15

    8-Methoxypsoralen (8-MOP) is a naturally occurring furanocoumarin with a variety of biological and pharmacological activities. The binding mechanism of 8-MOP to calf thymus DNA (ctDNA) at physiological pH was investigated by multi-spectroscopic techniques including UV–vis absorption, fluorescence, circular dichroism (CD) and Fourier transform infrared (FT-IR) spectroscopy along with DNA melting studies and viscosity measurements. The multivariate curve resolution-alternating least squares (MCR-ALS) chemometrics approach was introduced to resolve the expanded UV–vis spectral data matrix, and both the pure spectra and the equilibrium concentration profiles for the components (8-MOP, ctDNA and 8-MOP-ctDNA complex) in the system were successfully obtained to monitor the 8-MOP-ctDNA interaction. The results suggested that 8-MOP could bind to ctDNA via intercalation binding as evidenced by significant increases in melting and relative viscosity of ctDNA and competitive study using acridine orange (AO) as a fluorescence probe. The positive values of enthalpy and entropy change suggested that hydrogen bonds and van der Waals forces played a predominant role in the binding process. Further, FT-IR and CD spectra analysis indicated that 8-MOP preferentially bound to A–T base pairs with no major perturbation in ctDNA double helix conformation. Moreover, molecular docking was employed to exhibit the specific binding mode of 8-MOP to ctDNA intuitively. - Highlights: • The interaction processes of 8-MOP with ctDNA was monitored by MCR-ALS approach. • The binding mode of 8-MOP to ctDNA was an intercalation. • 8-MOP most likely bound to adenine and thymine base pairs of ctDNA. • Molecular docking illustrated the specific binding.

  4. Pseudopyronine B: A Potent Antimicrobial and Anticancer Molecule Isolated from a Pseudomonas mosselii.

    Science.gov (United States)

    Nishanth Kumar, S; Aravind, S R; Jacob, Jubi; Gopinath, Geethu; Lankalapalli, Ravi S; Sreelekha, T T; Dileep Kumar, B S

    2016-01-01

    In continuation of our search for new bioactive compounds from soil microbes, a fluorescent Pseudomonas strain isolated from paddy field soil of Kuttanad, Kerala, India was screened for the production of bioactive secondary metabolites. This strain was identified as Pseudomonas mosselii through 16S rDNA gene sequencing followed by BLAST analysis and the bioactive metabolites produced were purified by column chromatography (silica gel) and a pure bioactive secondary metabolite was isolated. This bioactive compound was identified as Pseudopyronine B by NMR and HR-ESI-MS. Pseudopyronine B recorded significant antimicrobial activity especially against Gram-positive bacteria and agriculturally important fungi. MTT assay was used for finding cell proliferation inhibition, and Pseudopyronine B recorded significant antitumor activity against non-small cell lung cancer cell (A549), and mouse melanoma cell (B16F10). The preliminary MTT assay results revealed that Pseudopyronine B recorded both dose- and time-dependent inhibition of the growth of test cancer cell lines. Pseudopyronine B induced apoptotic cell death in cancer cells as evidenced by Acridine orange/ethidium bromide and Hoechst staining, and this was further confirmed by flow cytometry analysis using Annexin V. Cell cycle analysis also supports apoptosis by inducing G2/M accumulation in both A549 and B16F10 cells. Pseudopyronine B treated cells recorded significant up-regulation of caspase 3 activity. Moreover, this compound recorded immunomodulatory activity by enhancing the proliferation of lymphocytes. The production of Pseudopyronine B by P. mosselii and its anticancer activity in A549 and B16F10 cell lines is reported here for the first time. The present study has a substantial influence on the information of Pseudopyronine B from P. mosselii as potential sources of novel drug molecule for the pharmaceutical companies, especially as potent antimicrobial and anticancer agent. PMID:27617005

  5. Effects of different cryoprotective agents on ram sperm morphology and DNAintegrity.

    Science.gov (United States)

    Nur, Z; Zik, B; Ustuner, B; Sagirkaya, H; Ozguden, C G

    2010-06-01

    This study investigates the effects of glycerol, 1,2 propanediol, sucrose, and trehalose on post-thaw motility, morphology, and genome integrity of Awassi ram semen. Ejaculates of thick consistency with rapid wave motion (>+++) and >70% initial motility were pooled. Sperm were diluted to a final concentration of 1/5 (semen/extender) in 0% cryoprotectant, 6% glycerol, 6% 1,2 propanediol, 62.5 mM sucrose or 62.5 mM trehalose using a two-step dilution method. The equilibrated semen was frozen in 0.25-ml straws. Semen samples were examined for sperm motility, defective acrosomes (FITC-Pisum sativum agglutinin (FITC PSA)), DNA integrity (acridine orange staining (AO)) and apoptotic activity (terminal deoxynucleotidyl transferase-mediated dUTP nick-end labeling (TUNEL) and Caspase-3 activity) at four time points: after dilution with extender A, after cooling to 5 degrees C, after equilibration and post-thaw. Freezing and thawing procedures (cooling at 5 degrees C, dilution, equilibration, and thawing) had negative effects on motility (P<0.001), acrosome integrity (P<0.001), and DNA integrity as determined by AO (P<0.001) and TUNEL (P<0.001) assays. There were positive correlations between sperm with defective acrosomes and apoptotic (AO- and TUNEL-positive) spermatozoa. In contrast, a significant negative correlation was found between sperm motility and defective acrosomes and AO- and TUNEL positivity (P<0.01). The cryopreservation process acts as an apoptotic inducer in ram semen; all cryoprotectants used in the present study allowed apoptosis to some extent, with negative effects on sperm morphology and DNA integrity. The glycerol group performed better than the propanediol, sucrose, trehalose, and control groups in terms of post-thaw sperm motility but not DNA integrity.

  6. Cytotoxicity, Antioxidant and Apoptosis Studies of Quercetin-3-O Glucoside and 4-(β-D-Glucopyranosyl-1→4-α-L-Rhamnopyranosyloxy)-Benzyl Isothiocyanate from Moringa oleifera.

    Science.gov (United States)

    Maiyo, Fiona C; Moodley, Roshila; Singh, Moganavelli

    2016-01-01

    Moringa oleifera, from the family Moringaceae, is used as a source of vegetable and herbal medicine and in the treatment of various cancers in many African countries, including Kenya. The present study involved the phytochemical analyses of the crude extracts of M.oleifera and biological activities (antioxidant, cytotoxicity and induction of apoptosis in-vitro) of selected isolated compounds. The compounds isolated from the leaves and seeds of the plant were quercetin-3-O-glucoside (1), 4-(β-D-glucopyranosyl-1→4-α-L-rhamnopyranosyloxy)-benzyl isothiocyanate (2), lutein (3), and sitosterol (4). Antioxidant activity of compound 1 was significant when compared to that of the control, while compound 2 showed moderate activity. The cytotoxicity of compounds 1 and 2 were tested in three cell lines, viz. liver hepatocellular carcinoma (HepG2), colon carcinoma (Caco-2) and a non-cancer cell line Human Embryonic Kidney (HEK293), using the MTT cell viability assay and compared against a standard anticancer drug, 5-fluorouracil. Apoptosis studies were carried out using the acridine orange/ethidium bromide dual staining method. The isolated compounds showed selective in vitro cytotoxic and apoptotic activity against human cancer and non-cancer cell lines, respectively. Compound 1 showed significant cytotoxicity against the Caco-2 cell line with an IC50 of 79 μg mL(-1) and moderate cytotoxicity against the HepG2 cell line with an IC50 of 150 μg mL(-1), while compound 2 showed significant cytotoxicity against the Caco- 2 and HepG2 cell lines with an IC50 of 45 μg mL(-1) and 60 μg mL(-1), respectively. Comparatively both compounds showed much lower cytotoxicity against the HEK293 cell line with IC50 values of 186 μg mL(-1) and 224 μg mL(-1), respectively. PMID:26428271

  7. Glucosylceramide modulates endolysosomal pH in Gaucher disease.

    Science.gov (United States)

    Sillence, Dan J

    2013-06-01

    GlcCer accumulation causes Gaucher disease where GlcCer breakdown is inhibited due to a hereditary deficiency in glucocerebrosidase. Glycolipids are endocytosed and targeted to the Golgi apparatus in normal cells but in Gaucher disease they are mistargeted to lysosomes. To better understand the role of GlcCer in endocytic sorting RAW macrophages were treated with Conduritol B-epoxide to inhibit GlcCer breakdown. Lipid analysis found increases in GlcCer led to accumulation of both triacylglycerol and cholesterol consistent with increased lysosomal pH. Ratio imaging of macrophages using both acridine orange and lysosensor yellow/blue to measure endolysosomal pH revealed increases in Conduritol B-epoxide treated RAW macrophages and Gaucher patient lymphoblasts. Increased endolysosomal pH was restricted to Gaucher lymphoblasts as no significant increases in pH were seen in Fabry, Krabbe, Tay-Sachs and GM1-gangliosidosis lymphoblasts. Substrate reduction therapy utilises inhibitors of GlcCer synthase to reduce storage in Gaucher disease. The addition of inhibitors of GlcCer synthesis to RAW macrophages also led to increases in cholesterol and triacylglycerol and an endolysosomal pH increase of up to 1 pH unit. GlcCer modulation appears specific since glucosylsphingosine but not galactosylsphingosine reversed the effects of GlcCer depletion. Although no acute effects on glycolipid trafficking were observed using bafilomycin A the results are consistent with a multistep model whereby increases in pH lead to altered trafficking via cholesterol accumulation. GlcCer modulates endolysosomal pH in lymphocytes suggesting an important role in normal lysosomes which may be disrupted in Gaucher disease. PMID:23628459

  8. Antiproliferation and apoptosis induction of paeonol in HepG2 cells

    Institute of Scientific and Technical Information of China (English)

    Shu-Ping Xu; Guo-Ping Sun; Yu-Xian Shen; Wei Wei; Wan-Ren Peng; Hua Wang

    2007-01-01

    AIM: To investigate the antiproliferative effect of paeonol (Pae) used alone or in combination with chemotherapeutic agents [cisplatin (CDDP), doxorubicin (DOX) and 5-fluorouracil (5-FU)] on human hepatoma cell line HepG2 and the possible mechanisms.METHODS: The cytotoxic effect of drugs on HepG2 cells was measured by 3-(4, 5-dimethylthiazol-2-yl)-2, 5-diphenyltetra-zolium bromide (MTT) assay.Morphologic changes were observed by acridine orange (AO) fluorescence staining. Cell cycle and apoptosis rate were detected by flow cytometry (FCM). Drug-drug interactions were analyzed by the coefficient of drug interaction (CDI).RESULTS: Pae (7.81-250 mg/L) had an inhibitory effect on the proliferation of HepG2 cells in a dose-dependent manner, with the IC50 value of (104.77 + 7.28) mg/L. AO fluorescence staining and FCM assays showed that Pae induced apoptosis and arrested cell cycle at S phase in HepG2 cells. Further, different extent synergisms were observed when Pae (15.63, 31.25, 62.5 mg/L) was combined with CDDP (0.31-2.5 mg/L), DOX (0.16-1.25 mg/L), or 5-FU (12.5-100 mg/L) at appropriate concentrations. The IC50 value of the three drugs decreased dramatically when combined with Pae (P <0.01). Of the three different combinations, the sensitivity of cells to drugs was considerably different.CONCLUSION: Pae had a significant growth-inhibitory effect on the human hepatoma cell line HepG2,which may be related to apoptosis induction and cell cycle arrest. It also can enhance the cytotoxicity of chemotherapeutic agents on HepG2 cells, and the S phase arrest induced by Pae may be one of the mechanisms of these interactions.

  9. Thymoquinone inhibits murine leukemia WEHI-3 cells in vivo and in vitro.

    Directory of Open Access Journals (Sweden)

    Landa Zeenelabdin Ali Salim

    Full Text Available BACKGROUND: Thymoquinone is an active ingredient isolated from Nigella sativa (Black Seed. This study aimed to evaluate the in vitro and in vivo anti-leukemic effects of thymoquinone on WEHI-3 cells. METHODOLOGY/PRINCIPAL FINDINGS: The cytotoxic effect of thymoquinone was assessed using an MTT assay, while the inhibitory effect of thymoquinone on murine WEHI-3 cell growth was due to the induction of apoptosis, as evidenced by chromatin condensation dye, Hoechst 33342 and acridine orange/propidium iodide fluorescent staining. In addition, Annexin V staining for early apoptosis was performed using flowcytometric analysis. Apoptosis was found to be associated with the cell cycle arrest at the S phase. Expression of Bax, Bcl2 and HSP 70 proteins were observed by western blotting. The effects of thymoquinone on BALB/c mice injected with WEHI-3 cells were indicated by the decrease in the body, spleen and liver weights of the animal, as compared to the control. CONCLUSION: Thymoquinone promoted natural killer cell activities. This compound showed high toxicity against WEHI-3 cell line which was confirmed by an increase of the early apoptosis, followed by up-regulation of the anti-apoptotic protein, Bcl2, and down-regulation of the apoptotic protein, Bax. On the other hand, high reduction of the spleen and liver weight, and significant histopathology study of spleen and liver confirmed that thymoquinone inhibited WEHI-3 growth in the BALB/c mice. Results from this study highlight the potential of thymoquinone to be developed as an anti-leukemic agent.

  10. Anfisurvivin Oligonucleofides Promote In Vitro Cisplatin Induction of Apoptosis in Osteosarcoma Cells

    Institute of Scientific and Technical Information of China (English)

    DaxinGao; HaiyanZhang; GangLu; TaoHuang

    2004-01-01

    OBJECTIVE To study the effect of antisense oligonucleotides (ASODN) of survivin, a member of a gene family of inhibitors of apoptosis, on the ability of cisplatin (DDP) to induce apoptosis in osteosarcoma cells.METHODS ASODN of survivin were synthesized and transfected into osteosarcoma OS-732 cells. The effects of ASODN alone, and with DDP as well as the effect of sense oligonucleotides (SODN) with and without DDP were examined. The reverse tanscriptase-polymerase chain reaction (RTPCR) was utilized to detect the expression of survivin mRNA in each group of OS-732 cells. The cells were examined by flow cytometry (FCM) and staining with acridine orange (AO) to determine the morphology of the cells and the level of apoptosis in each group. The MTT assay was used to estimate the condition of cell growth.RESULTS In comparing the cells transfected with ASODN with the control, SODN and DDP groups, we found that the expression of survivin mRNA was significantly decreased (P<0.01), and the level of apoptosis enhanced. ASODN-treated cells appeared atrophic with condensed chromatin, characteristic of typical apoptotic changes. Cell growth was relatively inhibited in the ASODN groups. Furthermore, the apoptotic index (AI) and cell growth-inhibition ratio (IR) were significantly higher in the ASODN+DDP group compared to each group treated with a single agent and compared to the SODN+DDP group.CONCLUSION ASODN can specifically inhibit the expression of the survivin gene in OS-732 cells, and correspondingly suppress survivin function, resulting in an increase in sensitivity to DDP and thus the ability of DDP to induce apoptosis in osteosarcoma cells.

  11. Synergistic effect of combining paeonol and cisplatin on apoptotic induction of human hepatoma cell lines

    Institute of Scientific and Technical Information of China (English)

    Shu-ping XU; Guo-ping SUN; Yu-xian SHEN; Wan-ten PENG; Hua WANG; Wei WE

    2007-01-01

    Aim: To investigate whether paeonol (Pae) has synergistic effects with cisplatin (CDDP) on the growth-inhibition and apoptosis-induction of human hepatoma cell lines HepG2 and SMMC-7721.Methods: The cytotoxic effect of drugs was measured by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl-tetrazolium bromide assay. The coefficient of drug interaction was used to analyze the nature of drug interactions. Morphological changes were observed by acridine orange fluo-rescence staining. Cell cycle and the apoptosis rate were detected by flow cytometry. Bcl-2 and Bax expression were assayed by immunohistochemical staining.Results: Pae or CDDP had antiproliferative effect on the 2 cell lines in a dose-dependent manner, with different sensitivities to drugs. More interestingly, a synergistic inhibitory effect on the viability of the 2 cell lines was observed after treatment with a combination of Pae (15.63, 31.25, and 62.5 mg/L) with various concentrations of CDDP. Further study showed typical mor-phological changes of apoptosis if the cells were exposed to the two agents for 24 h. The apoptotic rate of the cells with combination treatment was signifi-candy higher than that of cells treated with Pae or CDDP alone. The expression of Bcl-2 decreased and that of Bax increased in the treated groups, especially in the combination group, with the ratio of Bcl-2/Bax decreasing correspondingly.Additionally, a combination of Pae with CDDP resulted in a stronger S phase arrest, compared with Pae or CDDP alone.Conclusion: Pae, in combination with CDDP, had a significantly synergistic growth-inhibitory and apoptosis-in-ducing effect on the 2 human hepatoma cell lines, which may be useful in hepatoma treatment.

  12. Effects of spider Macrothele raven venom on cell proliferation and cytotoxicity in HeLa cells

    Institute of Scientific and Technical Information of China (English)

    Li GAO; Bao-en SHAN; Jing CHEN; Jiang-hui LIU; Da-xiang SONG; Bao-cheng ZHU

    2005-01-01

    Aim: To examine the effect of venom from the spider Macrothele raven on cell proliferation and cytotoxicity in human cervical carcinoma, HeLa cells. Methods:Morphological and biochemical signs of apoptosis appeared using acridine orange-ethidium bromide (AO/EB) staining. Marked morphological changes in HeLa cells after treatment with spider venom were observed using scanning electron microscopy (SEM) and transmission electron microscopy (TEM). Cell proliferation and cytotoxicity were determined by [methyl-3H] thymidine assay ([3H]TdR) and lactate dehydrogenase (LDH) release, respectively. DNA fragmentation and cell cycle distribution were monitored using flow cytometry. In addition, Western blot analysis was used to evaluate the level of caspase-3 expression. In vivo examination of the inhibition of the size of tumors in nude mice treated with spider venom was measured. Results: Marked morphological changes were observed using AO/EB staining, SEM and TEM assay. Spider venom at concentrations of 10-40 mg/L caused dose- and time-dependent inhibition of HeLa cell proliferation.The ratio of apoptosis and necrosis increased. The activity of caspase-3 was upregulated after spider venom treatment. In vivo study of tumor size revealed that tumors significantly decreased in size from controls to tumors treated for 3 weeks with spider venom (P<0.05). Conclusion: The inhibition of HeLa cells by the venom of the spider Macrothele raveni was carried out in three ways: induction of apoptosis, necrosis of toxicity damage and direct lysis. Spider venom is a novel anti-tumor material both in vitro and in vivo.

  13. Genotoxic Evaluation of Mexican Welders Occupationally Exposed to Welding-Fumes Using the Micronucleus Test on Exfoliated Oral Mucosa Cells: A Cross-Sectional, Case-Control Study

    Science.gov (United States)

    Jara-Ettinger, Ana Cecilia; López-Tavera, Juan Carlos; Zavala-Cerna, María Guadalupe; Torres-Bugarín, Olivia

    2015-01-01

    Background An estimated 800,000 people worldwide are occupationally exposed to welding-fumes. Previous studies show that the exposure to such fumes is associated with damage to genetic material and increased cancer risk. In this study, we evaluate the genotoxic effect of welding-fumes using the Micronucleus Test on oral mucosa cells of Mexican welders. Material and Methods We conducted a cross-sectional, matched case-control study of n = 66 (33 exposed welders, and 33 healthy controls). Buccal mucosa smears were collected and stained with acridine orange, observed under 100x optical amplification with a fluorescence lamp, and a single-blinded observer counted the number of micronuclei and other nuclear abnormalities per 2,000 observed cells. We compared the frequencies of micronuclei and other nuclear abnormalities, and fitted generalised linear models to investigate the interactions between nuclear abnormalities and the exposure to welding-fumes, while controlling for smoking and age. Results Binucleated cells and condensed-chromatin cells showed statistically significant differences between cases and controls. The frequency of micronuclei and the rest of nuclear abnormalities (lobed-nuclei, pyknosis, karyolysis, and karyorrhexis) did not differ significantly between the groups. After adjusting for smoking, the regression results showed that the occurrence of binucleated cells could be predicted by the exposure to welding-fumes plus the presence of tobacco consumption; for the condensed-chromatin cells, our model showed that the exposure to welding-fumes is the only reliable predictor. Conclusions Our findings suggest that Mexican welders who are occupationally exposed to welding-fumes have increased counts of binucleated and condensed-chromatin cells. Nevertheless, the frequencies of micronuclei and the rest of nuclear abnormalities did not differ between cases and controls. Further studies should shed more light on this subject. PMID:26244938

  14. Glutathione transferase mu 2 protects glioblastoma cells against aminochrome toxicity by preventing autophagy and lysosome dysfunction

    Science.gov (United States)

    Huenchuguala, Sandro; Muñoz, Patricia; Zavala, Patricio; Villa, Mónica; Cuevas, Carlos; Ahumada, Ulises; Graumann, Rebecca; Nore, Beston F; Couve, Eduardo; Mannervik, Bengt; Paris, Irmgard; Segura-Aguilar, Juan

    2014-01-01

    U373MG cells constitutively express glutathione S-transferase mu 2 (GSTM2) and exhibit 3H-dopamine uptake, which is inhibited by 2 µM of nomifensine and 15 µM of estradiol. We generated a stable cell line (U373MGsiGST6) expressing an siRNA against GSTM2 that resulted in low GSTM2 expression (26% of wild-type U373MG cells). A significant increase in cell death was observed when U373MGsiGST6 cells were incubated with 50 µM purified aminochrome (18-fold increase) compared with wild-type cells. The incubation of U373MGsiGST6 cells with 75 µM aminochrome resulted in the formation of autophagic vacuoles containing undigested cellular components, as determined using transmission electron microscopy. A significant increase in autophagosomes was determined by measuring endogenous LC3-II, a significant decrease in cell death was observed in the presence of bafilomycin A1, and a significant increase in cell death was observed in the presence of trehalose. A significant increase in LAMP2 immunostaining was observed, a significant decrease in bright red fluorescence of lysosomes with acridine orange was observed, and bafilomycin A1 pretreatment reduced the loss of lysosome acidity. A significant increase in cell death was observed in the presence of lysosomal protease inhibitors. Aggregation of TUBA/α-tubulin (tubulin, α) and SQSTM1 protein accumulation were also observed. Moreover, a significant increase in the number of lipids droplets was observed compared with U373MG cells with normal expression of GSTM2. These results support the notion that GSTM2 is a protective enzyme against aminochrome toxicity in astrocytes and that aminochrome cell death in U373MGsiGST6 cells involves autophagic-lysosomal dysfunction. PMID:24434817

  15. Genotoxic Evaluation of Mexican Welders Occupationally Exposed to Welding-Fumes Using the Micronucleus Test on Exfoliated Oral Mucosa Cells: A Cross-Sectional, Case-Control Study.

    Directory of Open Access Journals (Sweden)

    Ana Cecilia Jara-Ettinger

    Full Text Available An estimated 800,000 people worldwide are occupationally exposed to welding-fumes. Previous studies show that the exposure to such fumes is associated with damage to genetic material and increased cancer risk. In this study, we evaluate the genotoxic effect of welding-fumes using the Micronucleus Test on oral mucosa cells of Mexican welders.We conducted a cross-sectional, matched case-control study of n = 66 (33 exposed welders, and 33 healthy controls. Buccal mucosa smears were collected and stained with acridine orange, observed under 100x optical amplification with a fluorescence lamp, and a single-blinded observer counted the number of micronuclei and other nuclear abnormalities per 2,000 observed cells. We compared the frequencies of micronuclei and other nuclear abnormalities, and fitted generalised linear models to investigate the interactions between nuclear abnormalities and the exposure to welding-fumes, while controlling for smoking and age.Binucleated cells and condensed-chromatin cells showed statistically significant differences between cases and controls. The frequency of micronuclei and the rest of nuclear abnormalities (lobed-nuclei, pyknosis, karyolysis, and karyorrhexis did not differ significantly between the groups. After adjusting for smoking, the regression results showed that the occurrence of binucleated cells could be predicted by the exposure to welding-fumes plus the presence of tobacco consumption; for the condensed-chromatin cells, our model showed that the exposure to welding-fumes is the only reliable predictor.Our findings suggest that Mexican welders who are occupationally exposed to welding-fumes have increased counts of binucleated and condensed-chromatin cells. Nevertheless, the frequencies of micronuclei and the rest of nuclear abnormalities did not differ between cases and controls. Further studies should shed more light on this subject.

  16. Idarubicin induces mTOR-dependent cytotoxic autophagy in leukemic cells

    Energy Technology Data Exchange (ETDEWEB)

    Ristic, Biljana [Institute of Microbiology and Immunology, School of Medicine, University of Belgrade, Dr. Subotica 1, 11000 Belgrade (Serbia); Bosnjak, Mihajlo [Institute of Histology and Embryology, School of Medicine, University of Belgrade, Belgrade (Serbia); Arsikin, Katarina [Institute of Microbiology and Immunology, School of Medicine, University of Belgrade, Dr. Subotica 1, 11000 Belgrade (Serbia); Mircic, Aleksandar; Suzin-Zivkovic, Violeta [Institute of Histology and Embryology, School of Medicine, University of Belgrade, Belgrade (Serbia); Bogdanovic, Andrija [Clinic for Hematology, Clinical Centre of Serbia, School of Medicine, University of Belgrade, Belgrade (Serbia); Perovic, Vladimir [Institute of Microbiology and Immunology, School of Medicine, University of Belgrade, Dr. Subotica 1, 11000 Belgrade (Serbia); Martinovic, Tamara; Kravic-Stevovic, Tamara; Bumbasirevic, Vladimir [Institute of Histology and Embryology, School of Medicine, University of Belgrade, Belgrade (Serbia); Trajkovic, Vladimir, E-mail: vtrajkovic@med.bg.ac.rs [Institute of Microbiology and Immunology, School of Medicine, University of Belgrade, Dr. Subotica 1, 11000 Belgrade (Serbia); Harhaji-Trajkovic, Ljubica, E-mail: buajk@yahoo.com [Institute for Biological Research, University of Belgrade, Belgrade, Despot Stefan Blvd. 142, 11000 Belgrade (Serbia)

    2014-08-01

    We investigated if the antileukemic drug idarubicin induces autophagy, a process of programmed cellular self-digestion, in leukemic cell lines and primary leukemic cells. Transmission electron microscopy and acridine orange staining demonstrated the presence of autophagic vesicles and intracellular acidification, respectively, in idarubicin-treated REH leukemic cell line. Idarubicin increased punctuation/aggregation of microtubule-associated light chain 3B (LC3B), enhanced the conversion of LC3B-I to autophagosome-associated LC3B-II in the presence of proteolysis inhibitors, and promoted the degradation of the selective autophagic target p62, thus indicating the increase in autophagic flux. Idarubicin inhibited the phosphorylation of the main autophagy repressor mammalian target of rapamycin (mTOR) and its downstream target p70S6 kinase. The treatment with the mTOR activator leucine prevented idarubicin-mediated autophagy induction. Idarubicin-induced mTOR repression was associated with the activation of the mTOR inhibitor AMP-activated protein kinase and down-regulation of the mTOR activator Akt. The suppression of autophagy by pharmacological inhibitors or LC3B and beclin-1 genetic knockdown rescued REH cells from idarubicin-mediated oxidative stress, mitochondrial depolarization, caspase activation and apoptotic DNA fragmentation. Idarubicin also caused mTOR inhibition and cytotoxic autophagy in K562 leukemic cell line and leukocytes from chronic myeloid leukemia patients, but not healthy controls. By demonstrating mTOR-dependent cytotoxic autophagy in idarubicin-treated leukemic cells, our results warrant caution when considering combining idarubicin with autophagy inhibitors in leukemia therapy. - Highlights: • Idarubicin induces autophagy in leukemic cell lines and primary leukemic cells. • Idarubicin induces autophagy by inhibiting mTOR in leukemic cells. • mTOR suppression by idarubicin is associated with AMPK activation and Akt blockade.

  17. Autophagy blockade sensitizes the anticancer activity of CA-4 via JNK-Bcl-2 pathway

    Energy Technology Data Exchange (ETDEWEB)

    Li, Yangling; Luo, Peihua; Wang, Jincheng; Dai, Jiabin; Yang, Xiaochun; Wu, Honghai; Yang, Bo, E-mail: yang924@zju.edu.cn; He, Qiaojun, E-mail: qiaojunhe@zju.edu.cn

    2014-01-15

    Combretastatin A-4 (CA-4) has already entered clinical trials of solid tumors over ten years. However, the limited anticancer activity and dose-dependent toxicity restrict its clinical application. Here, we offered convincing evidence that CA-4 induced autophagy in various cancer cells, which was demonstrated by acridine orange staining of intracellular acidic vesicles, the degradation of p62, the conversion of LC3-I to LC3-II and GFP-LC3 punctate fluorescence. Interestingly, CA-4-mediated apoptotic cell death was further potentiated by pretreatment with autophagy inhibitors (3-methyladenine and bafilomycin A1) or small interfering RNAs against the autophagic genes (Atg5 and Beclin 1). The enhanced anticancer activity of CA-4 and 3-MA was further confirmed in the SGC-7901 xenograft tumor model. These findings suggested that CA-4-elicited autophagic response played a protective role that impeded the eventual cell death while autophagy inhibition was expected to improve chemotherapeutic efficacy of CA-4. Meanwhile, CA-4 treatment led to phosphorylation/activation of JNK and JNK-dependent phosphorylation of Bcl-2. Importantly, JNK inhibitor or JNK siRNA inhibited autophagy but promoted CA-4-induced apoptosis, indicating a key requirement of JNK-Bcl-2 pathway in the activation of autophagy by CA-4. We also identified that pretreatment of Bcl-2 inhibitor (ABT-737) could significantly enhance anticancer activity of CA-4 due to inhibition of autophagy. Taken together, our data suggested that the JNK-Bcl-2 pathway was considered as the critical regulator of CA-4-induced protective autophagy and a potential drug target for chemotherapeutic combination. - Highlights: • Autophagy inhibition could be a potential for combretastatin A-4 antitumor efficacy. • The JNK-Bcl-2 pathway plays a critical role in CA-4-induced autophagy. • ABT-737 enhances CA-4 anticancer activity due to inhibition of autophagy.

  18. Short-term weightlessness produced by parabolic flight maneuvers altered gene expression patterns in human endothelial cells.

    Science.gov (United States)

    Grosse, Jirka; Wehland, Markus; Pietsch, Jessica; Ma, Xiao; Ulbrich, Claudia; Schulz, Herbert; Saar, Katrin; Hübner, Norbert; Hauslage, Jens; Hemmersbach, Ruth; Braun, Markus; van Loon, Jack; Vagt, Nicole; Infanger, Manfred; Eilles, Christoph; Egli, Marcel; Richter, Peter; Baltz, Theo; Einspanier, Ralf; Sharbati, Soroush; Grimm, Daniela

    2012-02-01

    This study focused on the effects of short-term microgravity (22 s) on the gene expression and morphology of endothelial cells (ECs) and evaluated gravisensitive signaling elements. ECs were investigated during four German Space Agency (Deutsches Zentrum für Luft- und Raumfahrt) parabolic flight campaigns. Hoechst 33342 and acridine orange/ethidium bromide staining showed no signs of cell death in ECs after 31 parabolas (P31). Gene array analysis revealed 320 significantly regulated genes after the first parabola (P1) and P31. COL4A5, COL8A1, ITGA6, ITGA10, and ITGB3 mRNAs were down-regulated after P1. EDN1 and TNFRSF12A mRNAs were up-regulated. ADAM19, CARD8, CD40, GSN, PRKCA (all down-regulated after P1), and PRKAA1 (AMPKα1) mRNAs (up-regulated) provide a very early protective mechanism of cell survival induced by 22 s microgravity. The ABL2 gene was significantly up-regulated after P1 and P31, TUBB was slightly induced, but ACTA2 and VIM mRNAs were not changed. β-Tubulin immunofluorescence revealed a cytoplasmic rearrangement. Vibration had no effect. Hypergravity reduced CARD8, NOS3, VASH1, SERPINH1 (all P1), CAV2, ADAM19, TNFRSF12A, CD40, and ITGA6 (P31) mRNAs. These data suggest that microgravity alters the gene expression patterns and the cytoskeleton of ECs very early. Several gravisensitive signaling elements, such as AMPKα1 and integrins, are involved in the reaction of ECs to altered gravity.

  19. Selective oxidation with nanoporous silica supported sensitizers: An environment friendly process using air and visible light

    Energy Technology Data Exchange (ETDEWEB)

    Saint-Cricq, Philippe; Pigot, Thierry; Blanc, Sylvie [Institut des Sciences Analytiques et de Physicochimie pour l' Environnement et les Materiaux, Universite de Pau et des Pays de l' Adour, Helioparc-2 Av. du President Angot, F-64053 Pau Cedex 09 (France); Lacombe, Sylvie, E-mail: sylvie.lacombe@univ-pau.fr [Institut des Sciences Analytiques et de Physicochimie pour l' Environnement et les Materiaux, Universite de Pau et des Pays de l' Adour, Helioparc-2 Av. du President Angot, F-64053 Pau Cedex 09 (France)

    2012-04-15

    Highlights: Black-Right-Pointing-Pointer Photo-sensitizers were covalently grafted on silica matrices. Black-Right-Pointing-Pointer Grafted powdered silica was characterized by diffuse reflectance and emission spectroscopy. Black-Right-Pointing-Pointer Selective solvent-free photo-oxygenation was carried out with air under visible light. Black-Right-Pointing-Pointer Singlet generation and reactivity at the gas-solid interface was demonstrated. - Abstract: Transparent and porous silica xerogels containing various grafted photosensitizers (PSs) such as anthraquinone derivatives, Neutral Red, Acridine Yellow and a laboratory-made dicyano aromatics (DBTP) were prepared. In most cases, the xerogels were shown to be mainly microporous by porosimetry. The PSs were characterized in the powdered monoliths (form, aggregation, concentration) by electronic spectroscopy which also proved to be a useful tool for monitoring the material evolution after irradiation. These nanoporous xerogels were used as microreactors for gas/solid solvent-free photo-oxygenation of dimethylsulfide (DMS) using visible light and air as the sole reactant. All these PSs containing monoliths were efficient for gas-solid DMS oxidation, leading to sulfoxide and sulfone in varying ratios. As these polar oxidation products remained strongly adsorbed on the silica matrix, the gaseous flow at the outlet of the reactor was totally free of sulfide and odorless. The best results in term of yield and initial rate of degradation of DMS were obtained with DBTP containing xerogels. Moreover, as these materials were reusable without loss of efficiency and sensitizer photobleaching after a washing regeneration step, the concept of recyclable sensitizing materials was approved, opening the way to green process.

  20. Cardioprotective role of Syzygium cumini against glucose-induced oxidative stress in H9C2 cardiac myocytes.

    Science.gov (United States)

    Atale, Neha; Chakraborty, Mainak; Mohanty, Sujata; Bhattacharya, Susinjan; Nigam, Darshika; Sharma, Manish; Rani, Vibha

    2013-09-01

    Diabetic patients are known to have an independent risk of cardiomyopathy. Hyperglycemia leads to upregulation of reactive oxygen species (ROS) that may contribute to diabetic cardiomyopathy. Thus, agents that suppress glucose-induced intracellular ROS levels can have therapeutic potential against diabetic cardiomyopathy. Syzygium cumini is well known for its anti-diabetic potential, but its cardioprotective properties have not been evaluated yet. The aim of the present study is to analyze cardioprotective properties of methanolic seed extract (MSE) of S. cumini in diabetic in vitro conditions. ROS scavenging activity of MSE was studied in glucose-stressed H9C2 cardiac myoblasts after optimizing the safe dose of glucose and MSE by 3-(4,5-dimethyl-thiazol-2-yl)-2,5-diphenyl tetrazolium bromide. 2',7'-dichlorfluorescein diacetate staining and Fluorescence-activated cell sorting analysis confirmed the suppression of ROS production by MSE in glucose-induced cells. The intracellular NO and H2O2 radical-scavenging activity of MSE was found to be significantly high in glucose-induced cells. Exposure of glucose-stressed H9C2 cells to MSE showed decline in the activity of catalase and superoxide dismutase enzymes and collagen content. 4',6-diamidino-2-phenylindole, propidium iodide and 10-N-nonyl-3,6-bis (dimethylamino) acridine staining revealed that MSE protects myocardial cells from glucose-induced stress. Taken together, our findings revealed that the well-known anti-diabetic S. cumini can also protect the cardiac cells from glucose-induced stress. PMID:23512199