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Sample records for acridines

  1. Novel trisubstituted acridines as human telomeric quadruplex binding ligands

    Ungvarsky, J.; Plšíková, J.; Janovec, J.; Koval, J.; Mikeš, J.; Mikesová, L.; Harvanova, D.; Fedoročko, P.; Kristian, P.; Kašpárková, Jana; Brabec, Viktor

    2014-01-01

    Roč. 57, DEC 2014 (2014), s. 13-29. ISSN 0045-2068 Institutional support: RVO:68081707 Keywords : Braco 19 derivatives * Trisubstituted acridines * DNA binding Subject RIV: BO - Biophysics Impact factor: 2.152, year: 2014

  2. Methyltrioctylammonium chloride catalysed sonochemical synthesis of acridine diones

    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. Dependence of acridine adsorption on ligand hydration enthalpy

    Matzner, R.; Bales, R.C. (Univ. of Arizona, Tucson, AZ (United States). Dept. of Hydrology and Water Resources)

    1994-11-01

    The environmental fate of acridine is of concern because of its toxic and teratogenic properties. It is found in tobacco smoke, air pollution source effluents, recent lake sediments, wood preservative wastewater, wastewater treatment plant biosludge, and contaminated groundwater. The effect of the aqueous solution pH and ligand type (H[sub 2]PO[sup [minus

  4. Ion flotatsion photometric determination of tellurium with acridine and papaverine

    The ion flotation of tellirium (4) bromide complex with papaverine and acridine has been studied. Papeverine and its sublate with tellurium are found to possess surface-active properties. Acridine is not a surfactart. An ion-flotation photometric method has been elaborated to determine tellurium with papaverine in blister copper and in anodic copper without separation of the sample matrix from interfering ions. The tellurium sublate is flotated for five minutes at the air flow rate >= 25 ml/min. The Te samples contained (%): Se 0.01-0.05; Ni 0,03-0.07; Cu 98-99.6; Au 30-130 g/t, Ag 700-2000 g/t, also As, Sb, Sn. The relative standard deviation does not exceed 0.06

  5. Kinetic interaction in hydrogenitrogenation of quinoline and acridine

    Liquid fossil fuels contain numerous nitrogen compounds. During hydrodenitrogenation processes, these compounds for the active catalytic sites, with each compound affecting the kinetics of the other compounds. An understanding of the kinetic interaction is essential in using the results of model compound kinetics to predict the behavior of complex mixtures. In this work, the authors study the hydrodenitrogenation of quinoline and acridine in n-hexadecane over a commercial nickel-molybdenum catalyst in a batch autoclave reactor at 8.3 MPa (1200 psig) and 357-3900C. The reaction networks and kinetics of individual compounds were developed. These results confirm the existing knowledge of reaction networks for quinoline and acridine. Furthermore, their experiments show that formation for o-ethylaniline, o-toluidine and aniline are also important steps in quinoline denitrogenation. For total nitrogen removal, a dual site Langmuir-Hinshelwood type model considering separate sites for adsorption of hydrogen and nitrogen compounds give the best fit

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

    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

  7. Lyoluminescence induced by gamma-irradiated NaCl in aqueous acridine orange solution

    Kalkar, C.D.; Lala, Neeta (Poona Univ., Pune (India). Dept. of Chemistry)

    1990-01-01

    The emission spectrum of lyoluminescence of acridine orange was recorded on a Fuess spectrograph. A continuous emission band was observed from 500 to 640 nm. On resolution, it showed two-banded emission with peaks at 565 and 603 nm respectively. The colour centers released during dissolution of irradiated NaCl crystals in aqueous acridine orange solution react to produce excited dimer and trimer species of acridine orange. It is suggested that these excited species are responsible for the observed lyoluminescence emission spectrum of acridine orange. (author).

  8. Lyoluminescence induced by gamma-irradiated NaCl in aqueous acridine orange solution

    The emission spectrum of lyoluminescence of acridine orange was recorded on a Fuess spectrograph. A continuous emission band was observed from 500 to 640 nm. On resolution, it showed two-banded emission with peaks at 565 and 603 nm respectively. The colour centers released during dissolution of irradiated NaCl crystals in aqueous acridine orange solution react to produce excited dimer and trimer species of acridine orange. It is suggested that these excited species are responsible for the observed lyoluminescence emission spectrum of acridine orange. (author)

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

    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...

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

    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.

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

    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...

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

    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.

  13. Removal of acridine orange from water by graphene oxide

    Fiallos, D. Coello; Gómez, C. Vacacela; Usca, G. Tubón; Pérez, D. Cid; Tavolaro, P.; Martino, G.; Caputi, L. S.; Tavolaro, A.

    2015-02-01

    Dyes are usually used in textile manufacturing and are one of the major contaminations in water. Thus, from an environmental point of view, the removal of dyes is of great concern, and recent applications using carbon-based materials showed high adsorption ability. In this work we use graphene oxide (GO) produced by improved Hummer's method, for adsorption of acridine orange dye (AO) in water. GO is a material containing functional groups such as carboxyl, epoxy, ketone, and hydroxyl, that can adsorb cationic dyes. Factors such as initial concentration of dye, the amount of GO, temperature and contact time were evaluated. Results show that the adsorption equilibrium, with the removal of 40% of the dye, is reached in approximately 1 hour, and that the adsorption capacity increases at higher initial concentrations. The highest value of AO adsorbed was 229.8 mg/g equivalent to 92% removal percentage by using AO initial concentration 0.10 mg/mL. FT-IR analysis of GO with adsorbed AO shows changes in the stretching vibrational bands, which corroborate the AO/GO interaction due to the functional groups present in GO. Furthermore, AO adsorbed on GO does not desorb back into water. Our results show that GO is an effective adsorbent and could be used to treat effluents contaminated with dyes.

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

    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.

  15. Binding of fluorescent acridine dyes acridine orange and 9-aminoacridine to hemoglobin: Elucidation of their molecular recognition by spectroscopy, calorimetry and molecular modeling techniques.

    Chatterjee, Sabyasachi; Kumar, Gopinatha Suresh

    2016-06-01

    The molecular interaction between hemoglobin (HHb), the major human heme protein, and the acridine dyes acridine orange (AO) and 9-aminoacridine (9AA) was studied by various spectroscopic, calorimetric and molecular modeling techniques. The dyes formed stable ground state complex with HHb as revealed from spectroscopic data. Temperature dependent fluorescence data showed the strength of the dye-protein complexation to be inversely proportional to temperature and the fluorescence quenching was static in nature. The binding-induced conformational change in the protein was investigated using circular dichroism, synchronous fluorescence, 3D fluorescence and FTIR spectroscopy results. Circular dichroism data also quantified the α-helicity change in hemoglobin due to the binding of acridine dyes. Calorimetric studies revealed the binding to be endothermic in nature for both AO and 9AA, though the latter had higher affinity, and this was also observed from spectroscopic data. The binding of both dyes was entropy driven. pH dependent fluorescence studies revealed the existence of electrostatic interaction between the protein and dye molecules. Molecular modeling studies specified the binding site and the non-covalent interactions involved in the association. Overall, the results revealed that a small change in the acridine chromophore leads to remarkable alteration in the structural and thermodynamic aspects of binding to HHb. PMID:27077554

  16. Modification of surface properties of electrospun polyamide nanofibers by means of a perfluorinated acridine

    Bianco, Andrea; Iardino, Giacomo; Bertarelli, Chiara; Miozzo, Luciano; Papagni, Antonio; Zerbi, Giuseppe

    2007-08-01

    Polyamide 6 (PA6) nanofibers were prepared from formic acid solutions by using electrospinning technique. The fibers were smooth, defects free and with diameters smaller than 200 nm. Small amounts of a perfluorinated acridine were added as dopant to the feed solution to modify the wettability of the fibers. The effect of doping on the contact angle values is well apparent. The contact angle values go from 50° of pure PA6 to 120° when 6% of acridine is added. A comparison between fibers and films of pure and doped polyamide 6 was carried out in order to determine the effect of morphology on wettability. Thermal annealing near the Tg of the polymer promoted the segregation of the molecules to the surface, reaching contact angles of 131° with smaller amounts (4%) of acridine. The surface segregation was also promoted by time aging.

  17. Extraction-absorptiometric determination of chromium by acridine yellow in natural and waste waters

    In interaction of Cr(VI) anion with acridine basic dye - acridine yellow has been studied. The colored ionic associate could be extracted by dichlorethane: acetone (3:1) binary mixture in Ph 1 to 2 N hydrochloric acid solution. Optimal concentration of reagent is 1,83·10-3-2,94·10-3M. The celebration graph obeyed Beer's law over the range 0,625-10 mkg Cr/ml and the apparent molar absorptivity of the extract at 454 nm was 3,4·104±500 l mol-1cm1. The molar ratio between Cr(VI) anion and acridine yellow in ionic associate has been determined by method Asmuse which is (1:1). The influence of interfering elements on the determination of chromium has been studied. The elaborated methods has been applied for determination of Cr(VI) in natural and waste waters

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

    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.

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

    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.

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

    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

  1. Surface enhanced Raman scattering of new acridine based fluorophore adsorbed on silver electrode.

    Solovyeva, Elena V; Myund, Liubov A; Denisova, Anna S

    2015-10-01

    4,5-Bis(N,N-di(2-hydroxyethyl)iminomethyl)acridine (BHIA) is a new acridine based fluoroionophore and a highly-selective sensor for cadmium ion. The direct interaction of the aromatic nitrogen atom with a surface is impossible since there are bulky substituents in the 4,5-positions of the acridine fragment. Nevertheless BHIA molecule shows a reliable SERS spectrum while adsorbed on a silver electrode. The analysis of SERS spectra pH dependence reveals that BHIA species adsorbed on a surface can exist in both non-protonated and protonated forms. The adsorption of BHIA from alkaline solution is accompanied by carbonaceous species formation at the surface. The intensity of such "carbon bands" turned out to be related with the supporting electrolyte (KCl) concentration. Upon lowering the electrode potential the SERS spectra of BHIA do not undergo changes but the intensity of bands decreases. This indicates that the adsorption mechanism on the silver surface is realized via aromatic system of acridine fragment. In case of such an adsorption mechanism the chelate fragment of the BHIA molecule is capable of interaction with the solution components. Addition of Cd(2+) ions to a system containing BHIA adsorbed on a silver electrode in equilibrium with the solution leads to the formation of BHIA/Cd(2+) complex which desorption causes the loss of SERS signal. PMID:25956332

  2. Surface enhanced Raman scattering of new acridine based fluorophore adsorbed on silver electrode

    Solovyeva, Elena V.; Myund, Liubov A.; Denisova, Anna S.

    2015-10-01

    4,5-Bis(N,N-di(2-hydroxyethyl)iminomethyl)acridine (BHIA) is a new acridine based fluoroionophore and a highly-selective sensor for cadmium ion. The direct interaction of the aromatic nitrogen atom with a surface is impossible since there are bulky substituents in the 4,5-positions of the acridine fragment. Nevertheless BHIA molecule shows a reliable SERS spectrum while adsorbed on a silver electrode. The analysis of SERS spectra pH dependence reveals that BHIA species adsorbed on a surface can exist in both non-protonated and protonated forms. The adsorption of BHIA from alkaline solution is accompanied by carbonaceous species formation at the surface. The intensity of such "carbon bands" turned out to be related with the supporting electrolyte (KCl) concentration. Upon lowering the electrode potential the SERS spectra of BHIA do not undergo changes but the intensity of bands decreases. This indicates that the adsorption mechanism on the silver surface is realized via aromatic system of acridine fragment. In case of such an adsorption mechanism the chelate fragment of the BHIA molecule is capable of interaction with the solution components. Addition of Cd2+ ions to a system containing BHIA adsorbed on a silver electrode in equilibrium with the solution leads to the formation of BHIA/Cd2+ complex which desorption causes the loss of SERS signal.

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

    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

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

    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.

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

    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

  6. Dissipation of pH gradients in tonoplast vesicles and liposomes by mixtures of acridine orange and anions

    Acridine orange altered the response to anions of both ATP and inorganic pyrophosphate-dependent pH gradient formation in tonoplast vesicles isolated from oat (Avena sativa L.) roots and red beet (Beta vulgaris L.) storage tissue. When used as a fluorescent pH probe in the presence of I-, ClO3-, NO3-, Br-, or SCN-, acridine orange reported lower pH gradients than either quinacrine or [14C]methylamine. Acridine orange, but not quinacrine, reduced [14C]methylamine accumulation when NO3- was present indicating that the effect was due to a real decrease in the size of the pH gradient, not a misreporting of the gradient by acridine orange. Other experiments indicated that acridine orange and NO3- increased the rate of pH gradient collapse both in tonoplast vesicles and in liposomes of phosphatidylcholine and that the effect in tonoplast vesicles was greater at 24 degree C than at 12 degree C. It is suggested that acridine orange and certain anions increase the permeability of membranes to H+, possibly because protonated acridine orange and the anions form a lipophilic ion pair within the vesicle which diffuses across the membrane thus discharging the pH gradient

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

    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

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

    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.

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

    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 ...

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

    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...

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

    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.

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

    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.

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

    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.

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

    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

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

    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.

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

    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.

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

    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 ...

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

    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

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

    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.

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

    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

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

    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...

  2. Redox reactions of copper(II) upon electrospray ionization in the presence of acridine ligands with an amide side chain

    Tintaru, A.; Charles, L.; Milko, Petr; Roithová, J.; Schröder, Detlef

    2009-01-01

    Roč. 22, č. 3 (2009), s. 229-233. ISSN 0894-3230 R&D Projects: GA AV ČR KJB400550704; GA ČR GA203/08/1487 Institutional research plan: CEZ:AV0Z40550506 Keywords : acridine * copper * electrospray ionization * mass spectrometry * quinoline Subject RIV: CC - Organic Chemistry Impact factor: 1.602, year: 2009

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

    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.

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

    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

  5. Acridine orange staining and radiometric detection of microorganisms in blood cultures

    To determine whether acridine orange (AO) staining of blood cultures could be used as a substitute for blind subculture when used in conjunction with the BACTEC system (Johnston Laboratories, Inc., Towson, Md.), the two methods were compared on all BACTEC-negative specimens. Since blind subcultures were routinely performed in our laboratory on days 2 and 6 of incubation, AO staining was also performed on these days. Cultures which were BACTEC positive on day 1 of incubation were not included in the study. Of the 2,395 bottles tested after 2 days of incubation, 106 were subculture positive. Of these, 96 (90.6%) were also AO positive and BACTEC positive, 3 (2.8%) were AO positive and BACTEC negative, and 7 (6.6%) were AO negative and BACTEC positive. Of the 3,487 bottles tested on day 6 of incubation, 14 were subculture positive; 7 (50%) of these were AO positive and BACTEC positive, and seven were AO positive and BACTEC negative. Of the total of 10 culture-positive bottles missed by BACTEC, all were positive, and all 10 companion aerobic bottles were BACTEC positive. In both phases of the experiment, there was a total of only four false-positive AO stains. As a result of this investigation, we have substituted AO staining for blind subculturing of BACTEC-negative bottles

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

    *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.

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

    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.

  8. Diagnosis of malaria by acridine orange fluorescent microscopy in an endemic area of Venezuela

    Irene Bosch

    1996-02-01

    Full Text Available Fluorescent (acridine orange microscopical examination of capillary centrifuged blood (quantitative buffy coat [QBC®] analysis and Giemsa stained thick blood smears (GTS were compared for diagnosis of malaria in blood specimens from adults living in malaria transmission areas of the States of Bolivar and Amazonas in southeastern and south Venezuela, respectively. Of a total of 198 GTS examined, 95 subjects (48% showed parasitaemia. Among the 95 blood films with a positive GTS, 94 were judged positive by the QBC. However, positive QBC tubes were found in 29 out of 103 blood specimens with a negative GTS. Thus, relative to a GTS standard, the sensitivity and specificity of the QBC-test was 99.2% and 72%, respectively. Young trophozoites of Plasmodium vivax and P. falciparum could not be distinguished with certainty. It is confirmed that the QBC offers many advantages compared with the standard diagnosis of malaria parasites, specifically in the speed of staining and ease of interpretation. However, in places where P. falciparum and P. vivax occur, species and stage differentiation should be confirmed with the GTS.

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

    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...

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

    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

  11. Synthesis and antiproliferative activity of 9-benzylamino-6-chloro-2-methoxy-acridine derivatives as potent DNA-binding ligands and topoisomerase II inhibitors.

    Zhang, Wei; Zhang, Bin; Zhang, Wei; Yang, Ti; Wang, Ning; Gao, Chunmei; Tan, Chunyan; Liu, Hongxia; Jiang, Yuyang

    2016-06-30

    A series of 9-benzylamino acridine derivatives were synthesized as an extension of our discovery of acridine antitumor agents. Most of these acridine compounds displayed good antiproliferative activity with IC50 values in low micromole range and structure-activity relationships were studied. Topo I- and II- mediated relaxation studies suggested that all of our compounds displayed strong Topo II inhibitory activity at 100 μM, while only four exhibited moderate Topo I inhibitory activity. The typical compound 8p could penetrate A549 cancer cells efficiently. Compound 8p could intercalate within the double-stranded DNA structure and induce DNA damage. Moreover, compound 8p could induce A549 cells apoptosis through caspase-dependent intrinsic pathway and arrest A549 cells at the G2/M phase. PMID:27060757

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

    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+}.

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

    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...

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

    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

  15. Binding of 1-nitro-9- (3-dimethylaminopropylamino-acridine to the DNA of the apical meristem cells of adventitious onion (Allium cepa L. roots

    Danuta Antosiewicz

    2014-02-01

    Full Text Available It was established that one half of the ledakrin (I-nitro-9-(3-dimethylaminopropylamino-acridine bound to the DNA in the cells of the studied onion root tips (Allium cepa L., forms labile complexes with it, the remaining half is covalently attached to only one strand of the DNA. One molecule of covalently bound ledakrin falls on average to 104-2X104 pairs of bases.

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

    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...

  17. Short communication. Evaluation of a commercial kit based on acridine orange/propidium iodide to assess the plasma membrane integrity of ram sperm

    J. L. Yániz; I. Palacín; S. Vicente-Fiel; J. Gosalvez; C. López-Fernández; Santolaria, P.

    2013-01-01

    This study was designed to develop a semiautomatic computer assisted methodology to evaluate the membrane integrity of ram spermatozoa using a commercial kit based on acridine orange/propidium iodide (AO/PI) labelling and ImageJ software. The study was divided into two experiments. In the first trial, the new computer-assisted method was validated by mixing fresh semen samples with different volumes of freeze killed spermatozoa to determine proportions of damaged spermatozoa in the final samp...

  18. Thermodynamic properties of three-ring aza-aromatics. 1. Experimental results for phenazine and acridine, and mutual validation of experiments and computational methods

    Measurements leading to the calculation of thermodynamic properties for phenazine (Chemical Abstracts registry number [92-82-0]) in the ideal-gas state are reported. Experimental methods included adiabatic heat-capacity calorimetry, inclined-piston manometry, and combustion calorimetry. Thermodynamic properties for acridine (Chemical Abstracts registry number [260-94-6]) were reported previously and included those measured with adiabatic heat-capacity calorimetry, comparative ebulliometry, inclined-piston manometry, and combustion calorimetry. New measurement results for acridine reported here are densities determined with a vibrating-tube densimeter and heat capacities for the liquid phase at saturation pressure determined with a differential-scanning calorimeter (d.s.c.). All critical properties were estimated. Molar entropies for the ideal-gas state were derived for both compounds at selected temperatures. Independent calculations of entropies for the ideal-gas state were performed at the B3LYP/6-31+G(d, p) model chemistry for phenazine and acridine. These are shown to be in excellent accord with the calorimetric results. All results are compared with experimental property values reported in the literature.

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

    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)

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

    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

  1. [Determination of vitamin B12 concentration by fluorescence quenching with acridine orange-rhodamine 6G energy transfer system].

    Liu, Bao-sheng; Gao, Jing; Yang, Geng-liang

    2005-07-01

    An energy transfer technique between acridine orange (AO) and rhodamine 6G (R6G) was studied, and the optimum experimental conditions of energy transfer were defined. It was found that the effective energy transfer could occur between AO and R6G in the dodecylbenzene sodium sulfonate solution with Na2 HPO4-citric acid buffer solution at pH 5.0. The fluorescence intensity of AO-R6G system was diminished by vitamin B12 in an alkalescence medium. Based on the AO-R6G energy transfer system anovel fluorescence quenching method for the determination of vitamin B12 has been developed. Under optimal conditious, the linear range of calibration curves for the determination of vitamin B12 was 0-3.0 x 10(-5) mol x L(-1). The detection limits were 4.8 x 10(-7) mol x L(-1) for Vitamin B12. Among six times of determination, the relative standard deviation was 0.51%-0.64%, and the recovery was 98.40% -103.62%. The method features good recurrence, rapidity of reaction, good stability, and few interfering substances. It can be satisfactorily used in the determination of the injection content of vitamin B12. PMID:16241060

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

    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

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

    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

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

    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.

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

    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. - 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

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

    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

  7. 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.

    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

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

    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.

  9. 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.

    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

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

    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.

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

    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)

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

    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.

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

    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.

  14. 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

    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.

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

    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

  16. 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.

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

    2016-03-15

    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. PMID:26722674

  17. Short communication. Evaluation of a commercial kit based on acridine orange/propidium iodide to assess the plasma membrane integrity of ram sperm

    J. L. Yániz

    2013-03-01

    Full Text Available This study was designed to develop a semiautomatic computer assisted methodology to evaluate the membrane integrity of ram spermatozoa using a commercial kit based on acridine orange/propidium iodide (AO/PI labelling and ImageJ software. The study was divided into two experiments. In the first trial, the new computer-assisted method was validated by mixing fresh semen samples with different volumes of freeze killed spermatozoa to determine proportions of damaged spermatozoa in the final samples. The proportion of damaged spermatozoa in each sample determined by the automated procedure where highly correlated (R2=0.97, p<0.001 with the predicted theoretical values. In the second trial, the new method was compared with a previously validated method of membrane integrity assessment based on phase-contrast/propidium iodide (PH/PI methodology. Measurements by AO/PI were, on average, 4.0% larger than measurements by PH/PI (SD=7.02% and 1.79% smaller than measurements of sperm motility determined by CASA (SD=4.83. The AO/PI method was also more repeatable than the PH/PI. The double staining methodology coupled with the routine for image analysis allowing automatic determination of sperm membrane integrity means a reduction in processing time of 75% compared to the previously developed method using a single fluorochrome (3 vs 12 min on average if the incubation period was included. This facilitates its use when a large number of samples are analysed. Our results validate the new computer assisted method for assessing sperm membrane integrity in sheep. The new method developed, in addition to being a free tool, allows quick automatic determination of sperm viability, which facilitates its use in routine semen analysis.

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

    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

  19. 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

    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.

  20. 1H and 13C NMR spectra, structure and physicochemical features of phenyl acridine-9-carboxylates and 10-methyl-9-(phenoxycarbonyl)acridinium trifluoromethanesulphonates--alkyl substituted in the phenyl fragment.

    Krzymiński, K; Malecha, P; Zadykowicz, B; Wróblewska, A; Błażejowski, J

    2011-01-01

    The 1H and 13C NMR spectra of twelve phenyl acridine-9-carboxylates--alkyl-substituted in the phenyl fragment--and their 10-methyl-9-(phenoxycarbonyl)acridinium salts dissolved in CD3CN, CD3OD, CDCl3 and DMSO-d6 were recorded in order to examine the influence of the structure of these compounds and the properties of the solvents on chemical shifts and 1H-(1)H coupling constants. Experimental data were compared with 1H and 13C chemical shifts predicted at the GIAO/DFT level of theory for DFT(B3LYP)/6-31G** optimised geometries of molecules, as well as with values of 1H chemical shifts and 1H-(1)H coupling constants, estimated using ACD/HNMR database software to ensure that the assignment was correct. To investigate the relations between chemical shifts and selected structural or physicochemical characteristics of the target compounds, the values of several of these parameters were determined at the DFT or HF levels of theory. The HOMO and LUMO energies obtained at the HF level yielded the ionisation potentials and electron affinities of molecules. The DFT method provided atomic partial charges, dipole moments, LCAO coefficients of pz LUMO of selected C atoms, and angles reflecting characteristic structural features of the compounds. It was found that the experimentally determined 1H and 13C chemical shifts of certain atoms relate to the predicted dipole moments, the angles between the acridine and phenyl moieties, and the LCAO coefficients of the pz LUMO of the C atoms believed to participate in the initial step of the oxidation of the target compounds. The spectral and physicochemical characteristics of the target compounds were investigated in the context of their chemiluminogenic ability. PMID:21134782

  1. 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

    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.

  2. 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

    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)

  3. Acridine-intercalator based hypoxia selective cytotoxins

    Hypoxia selective cytotoxins of the general formula STR1 wherein n is from 1 to 5, and NO2 is in at least one of the 2, 4 or 5-positions of the imidazole are developed. Such compounds have utility as radiosensitizers and chemosensitizers. 9 figs

  4. 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

    SIXTO RAUL COSTAMAGNA

    2000-07-01

    Full Text Available 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 paralelamente por la técnica de Papanicolaou y por la coloración fluorescente con naranja de acridina. Los resultados mostraron que el PAP presenta una sensibilidad del 54,5% para la detección de T. vaginalis, validación efectuada frente a la coloración fluorescente con naranja de acridina, para una prevalencia de enfermedad en el grupo de mujeres estudiadas del 13,75% y un nivel de confianza del 95%. Para ensayos "en paralelo" con ambas coloraciones, el valor global de la prueba fue del 93,8%, con un valor predictivo del resultado negativo del 93,2%. Concluimos que si bien T. vaginalis es detectada en el PAP, éste no presenta sensibilidad significativamente elevada como para ser considerada como única prueba, debiéndose complementar siempre con una coloración fluorescente con naranja de acridina, u otra prueba de similar valorThe present study examined the validity of PAP staining, as used for vaginal cytology, against fluorescent staining with acridine orange in order to determine the value of a negative result of Trichomonas vaginalis obtained by a PAP smear. We examined eighty vaginal-secretion samples from 18- to 45-year-old female patients of the Hospital Municipal of the city of Bahía Blanca, Province of Buenos Aires, Argentina. The samples were stained in parallel by the PAP smear technique and the fluorescent staining technique with acridine orange described by Fripp in 1975. The results of our validation tests demonstrated that

  5. 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

    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

  6. Acridine nucleophilic displacement - possible culprit of acridine interaction with prion protein

    Šebestík, Jaroslav; Pavlíček, A.; Šafařík, Martin; Holada, K.; Hlaváček, Jan; Stibor, I.

    Praha : Ústav organické chemie a biochemie AV ČR, 2007 - (Slaninová, J.), s. 93-95 ISBN 978-80-86241-28-9. - (Collection Symposium Series. 9). [Biologically Active Peptides /10./. Praha (CZ), 11.04.2007-13.04.2007] R&D Projects: GA ČR GA203/07/1517 Institutional research plan: CEZ:AV0Z40550506 Keywords : aminoacridines * prion * quinacrine Subject RIV: CC - Organic Chemistry

  7. 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

    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

  8. Dependence of the Reactivity of Acridine on Its Substituents: A Computational and Kinetic Study

    Zawada, Zbigniew; Šebestík, Jaroslav; Šafařík, Martin; Bouř, Petr

    -, č. 34 (2011), s. 6989-6997. ISSN 1434-193X R&D Projects: GA ČR GA203/07/1517; GA ČR GAP208/11/0105; GA MŠk(CZ) LH11033 Grant ostatní: AV ČR(CZ) M200550902 Institutional research plan: CEZ:AV0Z40550506 Keywords : nitrogen heterocycles * kinetics * reaction mechanisms * transition states * density functional calculations Subject RIV: CF - Physical ; Theoretical Chemistry Impact factor: 3.329, year: 2011

  9. Divergent strategy for the synthesis of original dihydrobenzo- and dihydronaphtho-acridines.

    Solmont, Kathleen; Boufroura, Hamza; Souibgui, Amel; Fornarelli, Pauline; Gaucher, Anne; Mahuteau-Betzer, Florence; Ben Hassine, Béchir; Prim, Damien

    2015-06-14

    A straightforward access to numerous novel substituted dihydrobenzo- and dihydronaphthoacridines is described using a unique molecular platform in two key steps. A large range of carbon-based substituents such as aromatic, vinyl, alkynyl fragments through Pd-catalysed couplings has been installed. The molecular diversity is extended to the introduction of aza-heterocycles and further authorizes the installation of alkylamino chains by means of Cu-promoted C-N bond formation. Possible access to quinolinium salts is also described. The methodology revealed convenient preparation of a wide panel of molecules that display various rigidity/flexibility and lipophilic/hydrophilic balances. Finally, the influence of structural modulations on the photophysical properties of these novel architectures is also studied. It is noteworthy that styryl and alkynyl derivatives are emissive in water (ϕF up to 12%). PMID:25965873

  10. Acridin-9-yl exchange: A proposal for the action of some 9-aminoacridine drugs

    Šebestík, Jaroslav; Šafařík, Martin; Stibor, I.; Hlaváček, Jan

    2006-01-01

    Roč. 84, č. 6 (2006), s. 605-614. ISSN 0006-3525 R&D Projects: GA ČR(CZ) GA203/04/1421 Institutional research plan: CEZ:AV0Z40550506 Keywords : 9-aminoacridine * quinacrine * amino-amine-displacement * prion Subject RIV: CC - Organic Chemistry Impact factor: 2.480, year: 2006

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

    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.

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

    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...

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

    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

  14. 9,9-Dimethyl-12-(3-nitrophenyl-7,8,9,10,11,12-hexahydrobenz[a]acridin-11-one

    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.

  15. 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.

    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 ...

  16. 9,9-Dimethyl-12-(3-nitro­phen­yl)-7,8,9,10,11,12-hexa­hydro­benz[a]acridin-11-one

    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.

  17. 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

    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.

  18. Frequencies of micronuclei detected on Mytilus galloprovincialis by different staining techniques after treatment with zinc chloride.

    Majone, F; Beltrame, C; Brunetti, R

    1988-01-01

    The frequencies of micronuclei induced by ZnCl2 and detected on the gill tissue of the marine mussel Mytilus galloprovincialis with different staining techniques (acridine orange, gallocyanin chromallum, Feulgen, Giemsa) were compared. At least in the used system, the Feulgen and gallocyanin chromallum methods gave a frequency of micronuclei significantly lower than that obtained with the acridine orange and Giemsa techniques. No significant difference between the frequencies obtained with acridine orange and Giemsa was shown. So, though the acridine orange is surely the method which provides the more reliable data, in environmental screening works the Giemsa technique may be more suitable for its simplicity. PMID:2461512

  19. 10-[2-(Dimethyl­amino)eth­yl]-9-(4-methoxy­phen­yl)-3,3,6,6-tetra­methyl-3,4,6,7,9,10-hexa­hydro­acridine-1,8(2H,5H)-dione

    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.

  20. Fate of Carbamazepine during Water Treatment

    Kosjek, T.; Andersen, Henrik Rasmus; Kompare, Boris; Ledin, Anna; Heath, E.

    2009-01-01

    Seven transformation products of carbamazepine generated by at least one of three common water treatment technologies (W-radiation, oxidation with chlorine dioxide (ClO2), and biological treatment with activated Sludge) were identified by complementary use of ion trap, single quadrupole, and...... quadrupole-time-of-flight mass spectrometers. Acridine was formed during all of the three treatment processes, while acridine 9-carbaldehyde was identified as an intermediate during ClO2 oxidation. Further treatment of acridine with ClO2 produced 9-hydroxy-acridine, UV-treatment resulted in the formation of...... compared the treatment technologies according to the removal of carbamazepine and the production and decay of its transformation products. The most successful method for the removal of carbamazepine was UV treatment, while acridine and acridone were more susceptible to biological treatment. Therefore...

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

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

    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.

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

    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.

  4. Organic dyes removal using magnetically modified rye straw

    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

  5. Organic dyes removal using magnetically modified rye straw

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

    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

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

    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.

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

    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.

  9. DNA-Conjugated Organic Chromophores in DNA Stacking Interactions

    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...

  10. Synthesis of Thioacridine Derivatives Using Lawesson’s Reagent

    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.

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

    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.

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

    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.

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

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

    2004-01-01

    , 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...

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

    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.

  15. A novel immunoradiometric assay for human liver ferritin.

    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.

  16. FLOW CYTOMETRIC ANALYSIS OF EFFECTS OF 1,3-DINITROBENZENE ON RAT SPERMATOGENESIS

    Exposure of 100-d old rats to 1,3-dinitiobenzene (m-DNB) at dosages up to 48 mg/kg resulted in disruption of spermatogenesis as measured by flow cytometry (FCM) of acridine orange-stained sperm and testis cells. ne day (d 1) after a single exposure to 48 mg/kg m-DNB. CM measureme...

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

    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.

  18. Degranulation of mast cells and inhibition of the response to secretory agents by phototoxic compounds and ultraviolet radiation

    The symptoms of cutaneous phototoxicity from coal tar compounds and the nonsteroidal anti-inflammatory drug benoxaprofen are characterized by wheal and flare formation which is mediated by histamine released from dermal mast cells. Rat serosal mast cells were used as an in vitro model system to study the direct effect of phototoxic compounds on mast cell degranulation. The coal tar compounds studied included acridine and pyrene. Combined exposure of cells to acridine and UVA (320 to 400 nm) radiation caused mast cells to degranulate, as assayed by the release of [3H]serotonin. Maximum [3H]serotonin release (70 to 80%) was obtained with 50 microM acridine and 300 kJ/m2 UVA. Pyrene (25 microM), when photoexcited with UVB (280 to 360 nm) radiation, caused about 80% release of [3H]serotonin. No degranulation occurred with 20 microM benoxaprofen and UVB doses up to 7.2 kJ/m2. Trypan blue staining correlated well with degranulation caused by acridine plus UVA; however, with pyrene plus UVB there was greater [3H]serotonin release than dye uptake. Excitation of photosensitizers with doses of UV radiation that did not cause trypan blue staining suppressed degranulation of mast cells in response to chemical stimulation. Acridine, pyrene, and benoxaprofen in the presence of UV radiation inhibited the mast cells from responding to compound 48/80 or the calcium ionophore, chlortetracycline. Two other phototoxic compounds, chlorpromazine and deoxytetracycline, also abolished degranulation by compound 48/80. These findings indicate that phototoxic compounds: (1) cause degranulation in the presence of high doses of UV radiation; and (2) suppress degranulation of mast cells in response to secretory stimuli at doses of UV radiation that do not cause release of mediator

  19. Survival and activity of Streptococcus faecalis and escherichia coli in tropical freshwater

    The survival of Streptococcus faecalis 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. faecalis and E. coli decreased less than 1 log unit after 105 hours 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 hours, E. coli was more active than S. faecalis as measured by nucleic acid composition. In this tropical rain forest watershed, 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

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

    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.

  1. How prevalent are Plasmodium ovale and P. malariae in East Asia?

    Kawamoto, F; Liu, Q; Ferreira, M U; Tantular, I S

    1999-10-01

    Plasmodium ovale and Plasmodium malariae, two of the four human malaria parasites, are usually found at very low prevalence in East Asia, even in areas with intense malaria transmission. In this article, Fumihiko Kawamoto, Qing Liu, Marcelo Ferreira and Indah Tantular review data obtained in recent field surveys, using alternative diagnostic methods such as acridine orange staining and PCR-based methods, to evaluate the prevalence of these two malaria species in East Asia. They argue that these species might be much more prevalent in East Asia than reported previously. In addition, they discuss the implications of sequence variations found in the small subunit ribosomal RNA genes of the two species targeted by diagnostic PCR and compare morphological criteria for speciation of malaria parasites stained with Giemsa and acridine orange. PMID:10481157

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

    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

  3. Controlled evaluation of supplemented peptone and Bactec blood culture broths for the detection of bacteremia and fungemia.

    Reimer, L G; McDaniel, J D; Mirrett, S.; Reller, L B; Wang, W L

    1985-01-01

    Comparison of conventional blood culture media with newer formulations of Bactec media for radiometric detection are lacking. Therefore, we compared the yield and speed of detection of clinically important microorganisms with supplemented peptone broth (SPB) and Bactec aerobic (6B) and anaerobic (7C or 7D) broths in 7,627 blood samples from adult patients. Acridine orange stains from SPB, radiometric readings from Bactec, and routine subcultures from all bottles were done at the same time int...

  4. Fungi and Bacteria in or on Leaves of Eelgrass (Zostera marina L.) from Chesapeake Bay †

    Newell, Steven Y.

    1981-01-01

    Samples of green and brown leaves of eelgrass (Zostera marina L.) were incubated in seawater without an additional carbon source. Parallel leaf samples were used for acridine orange bacterial counting and water-soluble aniline blue estimation of fungal biovolume. The incubations produced no evidence that there is an eelgrass counterpart for the chytridialean symbiont which is very common in turtlegrass (Thalassia testudinum König). Sterile mycelium (i.e., living mycelium without identifiable ...

  5. Critical Evaluation of Techniques to Detect and Measure Cell Death – Study in a Model of UV Radiation of the Leukaemic Cell Line HL60

    Leite, Marina; Quinta‐Costa, Margarida; Leite, Pedro Simas; Guimarães, José Eduardo

    1999-01-01

    The reliability of eight distinct methods (Giemsa staining, trypan blue exclusion, acridine orange/ethidium bromide (AO/EB) double staining for fluorescence microscopy and flow cytometry, propidium iodide (PI) staining, annexin V assay, TUNEL assay and DNA ladder) for detection and quantification of cell death (apoptosis and necrosis) was evaluated and compared. Each of these methods detects different morphological or biochemical features of these two processes. The comparative analysis of th...

  6. Synthesis of 9-Aminoacridine Derivatives as Anti-Alzheimer Agents.

    Munawar, Rabya; Mushtaq, Nousheen; Arif, Sadia; Ahmed, Ahsaan; Akhtar, Shamim; Ansari, Sumaira; Meer, Sadia; Saify, Zafar S; Arif, Muhammad

    2016-05-01

    In the present study, some 9-aminoacridine derivatives have been synthesized by condensation of 9-aminoacridine with substituted phenacyl, benzoyl, and benzyl halides (RM1-RM6). Compounds were investigated for acetylcholinesterase and butyrylcholinesterase inhibition potential, considering these enzymes playing a key role in Alzheimer's disease. All derivatives showed better inhibition of enzymes than the standard galantamine, whereas except RM4, all exhibit better results than tacrine, a well-known acridine derivative used for the treatment of Alzheimer's disease. PMID:26385945

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

    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...

  8. Endovascular photodynamic therapy to prevent arterial restenosis

    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...

  9. ANTIPROLIFERATIVE AND APOPTOTIC EFFECTS OF THE ESSENTIAL OIL OF ORIGANUM ONITES AND CARVACROL ON HEP-G2 CELLS

    TOMSUK, Özlem; SİVAS, Hülya

    2011-01-01

    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...

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

    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

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

    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...

  12. Therapeutic study of proton beam in vascular disease animal models

    Proton beam radiation therapy is difficult to apply to animal model. When the cells with DNA damage in the irradiated zebrafish were stained with acridine orange, green fluorescent cell death spots were increased in trunk regions compared to non-irradiated control embryos. From this study, we found that proton radiation therapy can inhibit the blood vessel growth, which is probably induced in vivo in zebrafish embryos, and vascular endothelial cell proliferation.

  13. Chemical studies on the toxicology of the insecticide tamaron in the rat

    Non - Labelled O,S - dimethyl - phosphoramido - thiolate ( tamaron) was prepared by refluxing sulphur with phosphorus trichloride in the presence of anhydrous aluminium chloride to give the phosphoryl chloride(I), which upon reaction with absolute methanol in the presence of acridine and calcium oxide yielded O-methyl-phosphorodichloridothioate (II). Isomerisation of (II) gave S - methyl - phosphoro- dichlorido-thioate (III) .The latter reacted with absolute methanol and dry ammonia gas to give tamaron (IV)

  14. Identification of Escherichia coli DNA helicase IV with the use of a DNA helicase activity gel.

    Trieu, V N; McCarthy, D

    1989-01-01

    A DNA helicase activity gel was developed based on the assumption that DNA helicases could unwind double-stranded DNA in a polyacrylamide matrix. The production of single-stranded DNA was detected by staining the activity gel with acridine orange and visualizing the gel under long-wave UV light. The products of DNA helicase activities appeared as red bands within a green fluorescent background. A novel DNA helicase, called helicase IV, was detected in crude extracts of Escherichia coli with t...

  15. A role of the 9-aminoacridines and their conjugates in a life science

    Šebestík, Jaroslav; Hlaváček, Jan; Stibor, I.

    2007-01-01

    Roč. 8, č. 5 (2007), s. 471-483. ISSN 1389-2037 R&D Projects: GA ČR GA203/04/1421; GA ČR GA203/07/1517 Institutional research plan: CEZ:AV0Z40550506 Keywords : acridines * 9-aminoacridine * conjugates * nucleic acids * proteins Subject RIV: CC - Organic Chemistry Impact factor: 3.259, year: 2007

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

    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...

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

    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...

  18. Seasonal dimethylsulfoniopropionate (DMSP) variability in Dona Paula Bay

    Kumar, S.S.; Chinchkar, U.; Nair, S.; LokaBharathi, P.A.; Chandramohan, D.

    surfaces. Advances in Microbial Physiology 32, 53–85. Ginestet, P., Audic, J., Urbain, V., Block, J., 1998. Estimation of nitrifying bacterial activitiesby measuringoxygenuptakeinthepresenceof the metabolicinhibitors allylthioureaandazide.AppliedandEnvironmentalMicrobiology....2. Microbiological parameters Total bacterial numbers were enumerated by the acridine orange direct count (AODC) method (Hobbie et al.,1977). Samples were fixed immediately with formaldehyde (2% final concentra- tion). A fixed amount was filtered through 0.2 mm pore...

  19. Curcumin induces apoptosis in breast cancer cells and inhibits tumor growth in vitro and in vivo

    Lv, Zhi-Dong; Liu, Xiang-Ping; Zhao, Wei-Jun; Dong, Qian; Li, Fu-Nian; Wang, Hai-Bo; Kong, Bin

    2014-01-01

    Curcumin has shown therapeutic and/or adjuvant therapeutic effects on the treatment of some patients with breast cancer. However, its mechanisms of action are largely unknown. This study was designed to investigate its antitumor effect and underlying mechanisms in human breast cancer MDA-MB-231 and MCF-7 cells. The MTT assay was used to evaluate cell viability, and flow cytometry, acridine orange staining and transmission electron microscopy were used to detect apoptosis for cultured cells. T...

  20. Reaction of quinacrine with prion protein: treatment for Creutzfeldt-Jakob disease?

    Zawada, Zbigniew; Šebestík, Jaroslav; Šafařík, Martin; Březinová, Anna; Bouř, Petr; Hlaváček, Jan; Stibor, Ivan

    2010-01-01

    Roč. 104, č. 11 (2010), s. 1129-1129. ISSN 0009-2770. [Pokroky v organické, bioorganické a farmaceutické chemii /45./. 20.11.2010-22.11.2010, Nymburk] R&D Projects: GA ČR GA203/07/1517 Institutional research plan: CEZ:AV0Z40550506 Keywords : quinacrine * acridine displacement * prions * prevention of aggregation Subject RIV: CC - Organic Chemistry

  1. Reactive oxygen and nitrogen (ROS and RNS) species generation and cell death in tomato suspension cultures—Botrytis cinerea interaction

    Pietrowska, E.; Różalska, S.; Kaźmierczak, A.; Nawrocka, J.; Małolepsza, U.

    2014-01-01

    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 br...

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

    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 ...

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

    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)

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

    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

  5. Physical and chemical mutagenesis on a mycophagous nematode Aphelenchoides composticola (M.T. Franklin, 1957)

    Chemical mutagens as EMS, acriflavine, acridine, colchicine, nitrous acide and physical mutagens, such as X rays, have been used on the gonochoric mycophagous Nematode Aphelenchoides composticola. They show a nematicid activity due, to their toxicity on treated Nematodes and to the induction of lethal mutations affecting particularly early stages of gametogenesis. They produce abnormal strains dwarfs or giants (up to 25% of the population). Concentrations of chemical mutagens varying from 0.2 to 0.5% correspond to the optimal production of abnormalities. Similar results were obtained by irradiation near to 2000r. The action of the mutagens shows some differences: EMS and X rays generally produce dwarfs, whereas acriflavine, acridine, colchicine or nitrous acid induced only giants. Abnormal strains appear: in the F1, generation by X rays or acridine treatments; in the F2 or F3 generation by acriflavine, colchicine, nitrous acid or EMS action. The abnormal strains could be either variants or mutants and from these we select: four dwarfs B, C, D, E, induced by EMS 0.5% for 24 hours appearing in the F3 generation; or dwarf F induced by irradiation of 1500r appearing in the F1 generation. All these selected mutants are autosomal recessive single factors D and C controlled by two alleles of the some locus

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

    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).

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

    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

  8. Toluidine blue: the mode of photodynamic action in yeast cells

    Toluidine blue, a thiazine dye, was shown to have in vivo photodynamic activity through singlet oxygen (O21Δsub(g)) production. This was based mainly on the effective protection by N3- and the marked enhancement in D2O for the sensitized inactivation of yeast cells. The mode of the in vivo activity was, however, quite different from that of acridine orange, for which the singlet oxygen mechanism has also been proposed. The most characteristic feature in the toluidine blue-sensitization was the total lack of the induction of gene conversion (at trp 5), while the survival went down below 10%. The non-induction of genetic changes was confirmed at several pH's in the neutral region, whereas the inactivation was seen in parallel to the reported pH dependence of singlet oxygen production in vitro. Direct measurements by microspectrophotometry showed none of the toluidine blue was accumulated in the cell. It was also ascertained from acridine-sensitized induction of gene conversion that toluidine blue never interfered with the binding of acridine orange to cellular DNA. These findings suggested that the unique mode of photodynamic activity of toluidine blue is attributable to its action from outside of the cell. Furthermore, comparisons between the photodynamically treated cells (with toluidine blue) and non-treated cells with respect to the response to UV irradiation excluded certain cell functions relating to the expression of gene conversion from the possible damage sites. The photoreactivation process of UV induced gene conversion was not distributed by the pre-toluidine blue sensitization. In view of the foregoing results, the plasma membrane was tentatively suggested as the most likely site of damage. (author)

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

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

    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

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

    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

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

    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)

  13. Isolation of amoebae and Pseudomonas and Legionella spp. from eyewash stations.

    Paszko-Kolva, C.; Yamamoto, H.; Shahamat, M; Sawyer, T K; Morris, G.; Colwell, R. R.

    1991-01-01

    Forty eyewash units were sampled for protozoa, bacteria, and fungi. Total heterotrophic bacterial counts on nutrient agar and R2A agar (Difco Laboratories, Detroit, Mich.) ranged from 0 to 10(5) CFU/ml, with Pseudomonas spp. being the most frequently isolated. Total counts of 10(4) and 10(8) cells per ml were obtained with the acridine orange staining procedure. All samples were examined for Legionella spp. by direct fluorescent-antibody staining and by culturing on buffered charcoal-yeast ex...

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

    Pratiwi Sudarmono; Robert Utji; Kardono, Leonardus B. S.; Shirly Kumala

    2006-01-01

    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 cy...

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

    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.

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

    Y Padmanabha Reddy; K. B. Chandrasekhar; Mohammed Jaffar Sadiq

    2015-01-01

    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 antic...

  17. Okadaic acid inhibits cell multiplication and induces apoptosis in a549 cells, a human lung adenocarcinoma cell line

    Wang, Renjun; Lv, Lili; Zhao, Yunfeng; Yang, Nana

    2014-01-01

    This essay aims to research the effect of okadaic acid (OA) on A549 cell multiplication, and cell apoptosis induced by OA was observed by cell morphology. MTT assay, trypan blue exclusion test (TBET), Giemsa staining method and acridine orange (AO) fluorescence staining assay were applied. The results of cell survival evaluated by TBET and colorimetric assay with 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) showed: The number of A549 cells was decreased in a dose-depende...

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

    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...

  19. Visualization of DNA damage in individual cells

    A simple technique of micro-agarose gel electrophoresis has been developed to permit an evaluation of DNA damage in individual cells. Cells are embeded in agarose gel on microscope slides, lysed by detergents and then electrophoresed for a short time. In damaged cells, DNA migrated from the nuclei toward the anode, displaying 'comets' visualized by staining with a DNA-specific fluorochrome, acridine orange. The technique was applicable to quantifying DNA damage in individual cells exposed to Gy level of reactor radiation. (author)

  20. Visualization of ATP release in pancreatic acini in response to cholinergic stimulus. Use of fluorescent probes and confocal microscopy

    Sørensen, Christiane Elisabeth; Novak, Ivana

    2001-01-01

    receptors on pancreatic ducts. Thus, it was relevant to ask whether the upstream acini could be the source of releasable ATP and what the stimulus might be. We used freshly prepared rat pancreatic acini and applied conventional luminescence measurements of luciferin/luciferase reaction. As a new application...... partially overlapping with those marked by acridine orange and LysoTracker Red. In functional studies we show that native pancreatic acini release ATP in response to various stimuli but most importantly to cholinergic stimulation, a very likely physiological stimulus in this epithelium. In a close vicinity...

  1. Evaluation of sperm chromatin structure in boar semen

    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.

  2. Structure-activity relationship of indoloquinoline analogs anti-MRSA.

    Zhao, Min; Kamada, Tomonori; Takeuchi, Aya; Nishioka, Hiromi; Kuroda, Teruo; Takeuchi, Yasuo

    2015-12-01

    Indolo[3,2-b]quinoline analogs (3a-3s), 4-(acridin-9-ylamino) phenol hydrochloride (4), benzofuro[3,2-b]quinoline (3t), indeno[1,2-b]quinolines (3u and 3v) have been synthesized. Those compounds were found to exhibit anti-bacterial activity towards Methicillin-resistant Staphylococcus aureus (anti-MRSA activity). Structure-activity relationship studies were conducted that indoloquinoline ring, benzofuroquinoline ring and 4-aminophenol group are essential structure for anti-MRSA activity. PMID:26522949

  3. Photosensitized oxidation of DNA and its components

    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

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

    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...

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

    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...

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

    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.

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

    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.

  8. CONFIRMING LOCATION OF NITROGEN FIXING GENES ON PLASMIDS IN RHIZOBIUM ISOLATED FROM PISUM SATIVUM

    Balaji Hajare and Avinash Ade1

    2012-06-01

    Full Text Available To confirm the location of the nitrogen fixing genes whether on the plasmids or on the genomic DNA the Rhizobial isolates isolated from pea (Pisum sativum were treated with acridine orange with various concentrations and fixed nitrogen was estimated from the media in which these rhizobia were grown. There was no significant difference in between the cured and non cured strains of the Rhizobium which proved that the nitrogen fixing genes are not plasmid borne but these are located on the genomic DNA.

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

    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 ...

  10. Kinetics and mechanism of desulfurization and denitrogenation of coal-derived liquids. Eleventh quarterly report, December 21, 1977-March 20, 1978

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

    1978-05-20

    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, benzacridines, dibenzacridine, 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/. The results of the experiments are described. (LTN)

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

    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...

  12. Dye Transport across the Retinal Basement Membrane of the Blowfly Calliphora erythrocephala

    Weyrauther, E.; Roebroek, J.G.H.; Stavenga, D.G.

    1989-01-01

    In the blowfly, Calliphora erythrocephala, transport of dye into or out of the retina, following injection into the eye or thorax, was investigated, mainly by microspectrophotometry and fluorimetry. After injection into the eye, Phenol Red, Trypan Blue, Lucifer Yellow and 9-amino-acridine were transported out of the retina; Procion Yellow and Rhodamine-123 stayed in it. The time constants of this transport process were in the range 45-80 min at 23°C, depending on the dye. When Lucifer Yellow ...

  13. Microbiology of the Oil Fly, Helaeomyia petrolei

    Kadavy, Dana R.; Plantz, Bradley; Christopher A. Shaw; Myatt, Jill; Kokjohn, Tyler A.; Nickerson, Kenneth W.

    1999-01-01

    Helaeomyia petrolei larvae isolated from the asphalt seeps of Rancho La Brea in Los Angeles, Calif., were examined for microbial gut contents. Standard counts on Luria-Bertani, MacConkey, and blood agar plates indicated ca. 2 × 105 heterotrophic bacteria per larva. The culturable bacteria represented 15 to 20% of the total population as determined by acridine orange staining. The gut itself contained large amounts of the oil, had no observable ceca, and maintained a slightly acidic pH of 6.3 ...

  14. 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

    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.

  15. Integrative biological studies of anti-tumour agents

    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...

  16. Further in vivo studies on the participation of singlet oxygen in the photodynamic inactivation and induction of genetic changes in saccharomyces cerevisiae

    In vivo participation of singlet excited oxygen (1O2, 1Δsub(g)) in the photodynamic inactivation and induction of genetic changes (gene conversion) in acridine orange-sensitized yeast cells was investigated by using N3-, an efficient 1O2 quencher, and D2O, a known agent for the enhancement of the lifetime of 1O2. The addition of N3- protected the cells from both photodynamic actions. From an analysis of the concentration-dependent protection, about 80% of the induction of the genetic change is explainable on the basis of 1O2 mechanism. The quantitative estimation of the N3- protection in the inactivation was not possible because of the sigmoidal nature of the inactivation curve. The replacement of H2O with D2O during illumination was effective in enhancing the photodynamic inactivation but almost completely ineffective for the gene conversion induction. The deuterium effect with the cell system was clearly not as large as would be expected from in vitro experiments. This, however, could be explained from the kinetic consideration that natural quenchers of 1O2 in the cell would mask the deuterium effect. By experiments with different cell stages it was demonstrated that these two modifying effects were dependent on the intercellular reaction environment. The conclusion is that 1O2 must be the major intermediate responsible for the photodynamic actions in acridine orange-sensitized yeast cells. (author)

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

    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

  18. Effects of hyperoxia on microvascular cells in vitro

    Microvascular cells are most vulnerable to direct oxygen damage. Using an in vitro model system we have investigated the effect of elevated oxygen on the proliferation, morphology, and integrity of microvascular endothelial cells (EC) and pericytes. Cultivation of these cells at oxygen concentrations of 40% for 1 wk resulted in the inhibition of EC proliferation but had no effect on the growth of the pericytes. Similarly, hyperoxia induced a dramatic change in the shape of the EC, increasing their spread area by close to six-fold. Under the same conditions, the spread area of the pericytes was unaffected. To understand the effect of the hyperoxic treatment on the cells, the integrity of various membrane systems was assessed. 51Cr release was used to monitor plasma membrane integrity. There was no difference in chromium release by EC and pericytes over the 7 d of growth under normoxic and hyperoxic conditions. Mitochondrial integrity was examined by staining the cells with Rhodamine 123, which is selectively accumulated by the mitochondria. The staining pattern of the mitochondria of both EC and pericytes was altered by growth in the elevated oxygen. Finally, the lysosomes were visualized using acridine orange. The acridine orange staining pattern revealed enlarged and perinuclear lysosomes in the EC but no change in the pericyte lysosomal staining pattern. Thus, the cells of the microvasculature seem to be differentially affected by hyperoxia, a fact that may be significant in the etiology of reperfusion injury, ischemic disease, and pathologies associated with prematurity

  19. Synthesis and X-Ray Crystal Structure of Two Acridinedione Derivatives

    Dalbir Kour

    2014-01-01

    Full Text Available The two acridinedione derivatives 1 [3,3,6,6-tetramethyl-9-(4-methoxyphenyl-3,4,6,7,9,10-hexahydro-2H,5H-acridine-1,8-dione (C24H29NO3] and 2 [3,3,6,6-tetramethyl-9-(4-methylphenyl-3,4,6,7,9,10-hexa-hydro-2H,5H-acridine-1,8-dione (C24H29NO2] were synthesized and their crystal structures were determined by direct methods. The asymmetric unit of compound 1 contains two independent molecules. The 1,4-dihydropyridine (DHP ring adopts boat conformation in both 1 and 2. In 1 the dione rings exist in sofa conformation (for both the crystallographically independent molecules while the corresponding rings in 2 adopt half chair and sofa conformations, respectively. The crystal packing is stabilized by intermolecular N–H⋯O and C–H⋯O interactions in compound 1 and N–H⋯O interactions in compound 2.

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

    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.

  1. Toxicity of eight polycyclic aromatic compounds to red clover (Trifolium pratense), ryegrass (Lolium perenne), and mustard (Sinapsis alba).

    Sverdrup, Line E; Krogh, Paul Henning; Nielsen, Torben; Kjaer, Christian; Stenersen, Jørgen

    2003-12-01

    The effect of eight polycyclic aromatic compounds (PACs) on the seed emergence and early life-stage growth of three terrestrial plants (Sinapsis alba, Trifolium pratense and Lolium perenne) were studied in a greenhouse, using a Danish agricultural soil with an organic carbon content of 1.6%. After three weeks of exposure, seed emergence and seedling weight (fresh weight and dry weight) were determined. Exposure concentrations were verified with chemical analysis. The substances tested were four polycyclic aromatic hydrocarbons (fluoranthene, pyrene, phenanthrene and fluorene), the N-, S-, and O-substituted analogues of fluorene (carbazole, dibenzothiophene and dibenzofuran, respectively), and the quinoline representative acridine. Seedling growth was a far more sensitive endpoint than seed emergence for all substances. Concentrations estimated to give a 20% reduction of seedling fresh weight (EC20-values) ranged from 36 to 290 mgkg(-1) for carbazole, 43 to 93 mgkg(-1) for dibenzofuran, 37 to 110 mgkg(-1) for dibenzothiophene, 140 to 650 mgkg(-1) for fluoranthene, 55 to 380 mgkg(-1) for fluorene, 37 to 300 mgkg(-1) for phenanthrene, and 49 to 1300 mgkg(-1) for pyrene. For acridine, no toxicity was observed within the concentration range tested (1-1000 mgkg(-1)). As illustrated by the EC20-values, there was a rather large difference in sensitivity between the species, and T. pratense was the most sensitive of the species tested. PMID:14505722

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

    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

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

    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.

  4. Synthesis, DNA Binding and Antitumor Evaluation of Styelsamine and Cystodytin Analogues

    Hugo K. H. Fong

    2013-01-01

    Full Text Available A series of N-14 sidechain substituted analogues of styelsamine (pyrido[4,3,2-mn]acridine and cystodytin (pyrido[4,3,2-mn]acridin-4-one alkaloids have been prepared and evaluated for their DNA binding affinity and antiproliferative activity towards a panel of human tumor cell lines. Overall it was found that styelsamine analogues were stronger DNA binders, with the natural products styelsamines B and D having particularly high affinity (Kapp 5.33 × 106 and 3.64 × 106 M−1, respectively. In comparison, the cystodytin iminoquinone alkaloids showed lower affinity for DNA, but were typically just as active as styelsamine analogues at inhibiting proliferation of tumor cells in vitro. Sub-panel selectivity towards non-small cell lung, melanoma and renal cancer cell lines were observed for a number of the analogues. Correlation was observed between whole cell activity and clogP, with the most potent antiproliferative activity being observed for 3-phenylpropanamide analogues 37 and 41 (NCI panel average GI50 0.4 μM and 0.32 μM, respectively with clogP ~4.0–4.5.

  5. Rapid diagnosis of malaria by fluorescent microscopy with light microscope and interface filter

    The present study is planned to compare acridine orange (A.O) staining with Giemsa staining by using light microscopy with IF and also with fluorescent microscopy for detection of parasites in peripheral blood of patients suffering from clinically suspected cases of malaria. 200 patients with fever and shivering were included. General investigations like Hb, TLC and platelets were done by sysmex K-1000. Thin and thick blood films were made and stained according to protocol given i.e. by Giemsa and AO stains and slides were examined by different microscopes i.e. light microscope, light microscope with IFS and fluorescent microscope. Out of 200 subjects, 170 (85%) patients showed positive parasitaemia and 30 (15%) subjects were negative for malaria parasites. fib, TLC and platelets were reduced when comparing with MP negative cases. IFS microscope with acridine orange staining showed early detection of malaria parasites by counting fewer fields as compared to light microscopy with Giemsa stains. Time consumed for detection of parasites was also significantly reduced in IFS microscope by using AO stains. (author)

  6. Genotoxic and hematological parameters in Colossoma macropomum (Pisces, Serrasalmidae) as biomarkers for environmental impact assessment in a protected area in northeastern Brazil.

    Fortes Carvalho Neta, Raimunda Nonata; Pinheiro Sousa, Débora Batista; de Macêdo Sobrinho, Inaldo Carvalho; Yarbrough Horton, Emily; da Silva de Almeida, Zafira; Tchaicka, Lígia; de Sousa, Alana Lislea

    2015-10-01

    Genotoxic and hematological parameters in tambaqui (Colossoma macropomum), a native freshwater fish, were used as biomarkers to assess exposure to environmental stressors within the Maracanã Protected Area of Maranhão State, Brazil. Fish were sampled at two sites-Serena Lagoon (control) and Ambude River-on four occasions (dry and rainy season), and biometric data (length and weight) recorded and blood collected from all fish for analysis. Erythrocyte indices-mean corpuscular volume, mean corpuscular hemoglobin, and mean corpuscular hemoglobin concentration-were calculated. Blood samples were stained with Giemsa and acridine orange, and microscopically examined for micronuclei and morphological nuclear abnormalities. Micronuclei were observed in fish from both sites, although the frequency was significantly higher in fish from the Ambude River and morphological nuclear abnormalities were only observed in fish from the Ambude River. More morphological nuclear abnormalities and a larger number of micronuclei were observed in erythrocytes stained with acridine orange compared with those stained with Giemsa. On average, erythrocyte indices were lower in fish from the Ambude River than from the Serena Lagoon. The results confirm that genotoxic and hematological parameters in C. macropomum can be used as indicators of environmental health and could be valuable tools for monitoring environmental conditions within protected areas. PMID:26062456

  7. Genotoxicity of non-covalent interactions: DNA intercalators

    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

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

    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...

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

    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

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

    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

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

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

    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

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

    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.

  14. Regioselectivity and Tautomerism of Novel Five-Membered Ring Nitrogen Heterocycles Formed via Cyclocondensation of Acylthiosemicarbazides

    Karel D. Klika

    2008-03-01

    Full Text Available A series of 1-acyl-4-phenyl/(acridin-9-ylthiosemicarbazides 3, including fournew compounds, were prepared in order to study substituent effects on cyclizationreactions with oxalyl chloride (producing imidazolidine-4,5-diones 4, dimethylacetylenedicarboxylate (to give thiazolidin-4-ones 7 and 8 and autocondensation underalkaline conditions (to yield 1,2,4-triazoles 9. A positional isomer, 10 of compound 3f wasalso prepared. Altogether, twenty new compounds characterized and identified by IR, UV,1H, 13C and 2D NMR and quantum chemical calculations are described. The tautomerismof the products and regioselectivity of the reactions were evaluated. Compounds 3f−h,3h·2HCl, 7b,d and 10 were screened for cytotoxic activity against the L1210 leukemia cellline and all compounds, except for 3f, exhibited promising inhibitions of cell growth.

  15. New staining methods for yeast like fungi under special consideration of human pathogenic fungi

    Paulitsch-Fuchs, Astrid; Treiber, Fritz; Grasser, Erik; Buzina, Walter; Rosker, Christian

    2010-11-01

    A new method for in-cellular staining of yeast like fungi with Oregon Green and SYTOX Green is presented enabling their detection as well as the observation of cellular details via confocal laser scanning microscopy. Fluorochromes play an important role in many scientific disciplines including medicine, cell biology and botany. For the visualisation of fungal cell walls Calcofluor White is the flourochrome of choice. The necessity of an UV laser for its excitation makes it unpracticable for daily use. Safranin O, DAPI, 2NBDG, Ethidium Bromide and Acridin-orange are commonly used stains for nuclei in fugal microscopy. The attention was given to the possibility of using the differences in staining patterns to distinguish certain pathogenic yeast species e.g. Candida albicans and Candida krusei. Our results show that high quality microscopy of yeast like organisms can readily be achieved by the use of two suitable fluorochromes.

  16. Mitotic recombination induced by chemical and physical agents in the yeast Saccharomyces cerevisiae

    The treatment of diploid cultures of yeast with ultraviolet light (uv), γ-rays, nitrous acid (na) and ethyl methane sulphonate (ems) results in increases in cell death, mitotic gene conversion and crossing-over. Acridine orange (ao) treatment, in contrast, was effective only in increasing the frequency of gene conversion. The individual mutagens were effective in the order uv>na>γ-rays>ao>ems. Prior treatment of yeast cultures in starvation medium produced a significant reduction in the yield of induced gene conversion. The results have been interpreted on the basis of a general model of mitotic gene conversion which involves the post-replication repair of induced lesions involving de novo DNA synthesis without genetic exchange. In contrast mitotic crossing-over appears to involve the action of a repair system independent from excision or post-replication repair which involves genetic exchange between homologous chromosomes

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

    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)

  18. Ecotoxicity of carbamazepine and its UV photolysis transformation products

    Donner, E.; Kosjek, T.; Qualmann, Signe;

    2013-01-01

    considerably more toxic than carbamazepine itself may be produced during UV-treatment of wastewater effluents and/or photo-induced degradation of carbamazepine in natural waters. This study highlights the need to consider mixture toxicity and the formation and persistence of toxicologically relevant......Carbamazepine, an anti-epileptic pharmaceutical agent commonly found in wastewater, is highly recalcitrant to standard wastewater treatment practices. This study investigated the mixture toxicity of carbamazepine transformation products formed during ultraviolet (UV) photolysis using three standard...... treatment period, together with concurrent increases in acridine and acridone concentrations. Ecotoxicity was shown to increase in parallel with carbamazepine degradation indicating that the mixture of degradation products formed was more toxic than the parent compound, and all three ecotoxicity endpoints...

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

    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

  20. Mechanism and In Vivo Evaluation: Photodynamic Antibacterial Chemotherapy of Lysine-Porphyrin Conjugate.

    Xu, Zengping; Gao, Yuxiang; Meng, Shuai; Yang, Baochen; Pang, Liyun; Wang, Chen; Liu, Tianjun

    2016-01-01

    Lysine-porphyrin conjugate 4i has potent photosensitive antibacterial effect on clinical isolated bacterial strains such as Methicillin-resistant Staphylococcus aureus (MRSA), Escherichia coli, and Pseudomonas aeruginosa. The mechanism of photodynamic antibacterial chemotherapy of 4i (4i-PACT) in vitro and the treatment effect in vivo was investigated in this paper. Atomic force microscopy (AFM) revealed that 4i-PACT can effectively destroy membrane and wall of bacteria, resulting in leakage of its content. This was confirmed by dual fluorescent staining with acridine orange/ethidium bromide and measuring materials absorption at 260 nm. Agarose gel electrophoresis measurement showed that 4i-PACT can damage genomic DNA. Healing of wound in rat infected by mixed bacteria showed that the efficiency of 4i-PACT is dependent on the dose of light. These results showed that 4i-PACT has promising bactericidal effect both in vitro and in vivo. PMID:26973620

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

    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

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

    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

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

    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

  4. Soil microbial toxicity of eight polycyclic aromatic compounds: effects on nitrification, the genetic diversity of bacteria, and the total number of protozoans

    Sverdrup, Line Emilie; Ekelund, Flemming; Krogh, Paul Henning;

    2002-01-01

    measured by denaturant gradient gel electrophoresis. In general, only weak effects at very high concentrations were found for the protozoans. However, for acridine, protozoan numbers were reduced at lower concentrations than those that affected the nitrification process, that is, with a 5% reduction at 380...... mg/kg. For effects on nitrification, toxicity (NOEC values) expressed as soil pore-water concentrations (log10(micromol/L)) showed a significant inverse relationship with lipophilicity (log octanol-water partition coefficient) of the substances (r2 = 0.69, p = 0.011, n = 8). This finding could...... indicate that the toxicity of substances similar to those tested might be predicted by a quantitative structure-activity relationship with lipophilicity as the predictor variable....

  5. Preparation and photo-catalytic behavior of conjugated polymers based on paper-making wastewater.

    Feng, Libang; Qiang, Xiaohu; Shi, Xueting

    2009-08-01

    Based on alkaline paper-making wastewater, a polymer catalyst (FQ) was prepared and characterized by FTIR, ESR and element analysis techniques. The results show that the catalyst has conjugated structure and the conjugate degree increases after heat treatment. The catalyst has quite high photo-catalytic activity, which was verified by the fact that the simulated dyeing wastewater containing methylene blue (MB) or acridine orange (AO) can be degraded completely in 20 minutes under natural light using FQ as the photo-catalyst. Therefore, the synthetic dyeing wastewater can be disposed of using the materials coming from paper-making wastewater. It is a very promising method to treat one kind of wastewater with the materials from another kind of wastewater. PMID:20183197

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

    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.

  7. Evaluation of neurocytotoxic effects after exposure to Fe nuclei on developing optic tectum of Medaka, Oryzias latipes

    The aim of this study is to investigate neurocytotoxic effects of high linear energy transfer (LET) radiation on developing brain using medaka embryos. In our previous study, medaka embryos at stage 28 which were irradiated with lower dose than lethal dose 50% (LD50), were observed radiation induced apoptosis in the optic tectum transiently. To assess these radiation-induced apoptosis quantitatively, we developed a rapid and simple method using acridine orange (AO) in whole mount specimens. By using this method, we could evaluate the neurocytotoxic effects of x-rays on developing brain using medaka embryos. In this study, we tried to evaluate the neuryocyototoxic effects of Fe nuclei by using this AO-staining method. As a result, relative biological effectiveness values for radiation-induced apoptosis in central nervous system (CNS) were about 7 for exposure to Fe nuclei. (author)

  8. Time-resolved spectroscopy and fluorescence resonance energy transfer in the study of excimer laser damage of chromatin

    Radu, L.; Mihailescu, I.; Radu, S.; Gazdaru, D.

    2007-09-01

    The analysis of chromatin damage produced by a 248 nm excimer laser radiation, for doses of 0.3-3 MJ/m 2 was carried out by time-resolved spectroscopy and fluorescence resonance energy transfer (FRET). The chromatin was extracted from a normal and a tumoral tissue of Wistar rats. The decrease with laser dose of the relative contribution of the excited state lifetimes of ethidium bromide (EtBr) bounded to chromatin constitutes an evidence of the reduction of chromatin deoxyribonucleic acid (DNA) double-strand structure. FRET was performed from dansyl chloride to acridine orange, both coupled to chromatin. The increase of the average distance between these ligands, under the action of laser radiation, reflects a loosening of the chromatin structure. The radiosensitivity of tumor tissue chromatin is higher than that of a normal tissue. The determination of the chromatin structure modification in an excimer laser field can be of interest in laser therapy.

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

    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...

  10. A novel pH-sensitive polymeric fluorescent probe: Synthesis, characterization and optical properties

    A novel initiator containing proflavine derivative moiety, 3,6-dibromo-isobutyramide acridine (DIA), was synthesized and initiated the atom transfer radical polymerization (ATRP) of methyl methacrylate (MMA). A water-soluble monomer, N,N-dimethylacrylamide (DMAA) was also initiated by DIA for comparison. The obtained fluorescent polymers, PMMA-DIA and PDMAA-DIA, were characterized by 1H NMR, GPC and TGA. The emission spectra of the fluorescent polymers exhibit obvious changes in color and fluorescence intensity along with pH varied in range of 3.0-9.0. In addition, the obtained polymers present good film-forming capacity and the films also have a high quantum yield and pH response. Both oil-soluble PMMA-DIA and water-soluble PDMAA-DIA have steady optical and chemical properties by containing proflavine moiety in the main chain.

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

    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

  12. Exploration for the multi-effect of cardamom in's resistance to multiple myeloma.

    Zhihua, Zhao; Jianping, Yang; Miaomiao, Sun; Kuisheng, Chen

    2014-11-01

    This paper aimed to probe the cardamom in effect on the viability, proliferation, apoptosis and periodic function of the multiple myeloma, and explore its mechanism. We used CCK-8 method to evaluate the effect of cardamom in on the viability of PBMNCs (Persom Blood Mononuclear Normal Cells). EdU can test the influence of small cell proliferation. We used the method of PI single-staining flow cytometry, in order to test the influence of tumor cell cycle. AO (Acridine Orange), EB (Ethidium Bromide) double staining fluorescene microscope was applied to observe the influence of tumor apoptotic morphology. It can be concluded that cardamom in can inhibit the viability and proliferation of MM (Multiple Myeloma) cells and cardamom in is the anti-myeloma drug with strong viability. PMID:25410063

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

    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.

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

    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.

  15. Spectrofluorimetric determination of second critical micellar concentration of SDS and SDS/Brij 30 systems.

    Romani, Ana Paula; Machado, Antonio Eduardo da Hora; Hioka, Noboru; Severino, Divinomar; Baptista, Mauricio S; Codognoto, Lúcia; Rodrigues, Maira R; de Oliveira, Hueder Paulo Moisés

    2009-03-01

    Potentially useful stead-state fluorimetric technique was used to determine the critical micellar concentrations (CMC(1) and CMC(2)) for two micellar media, one formed by SDS and the other by SDS/Brij 30. A comparative study based on conductimetric and surfacial tension measurements suggests that the CMC(1) estimated by the fluorimetric method is lower than the value estimated by these other techniques. Equivalent values were observed for SDS micelles without Brij 30 neutral co-surfactant. The use of acridine orange as fluorescent probe permitted to determine both CMC(1) and CMC(2). Based on it an explanation on aspects of micelle formation mechanism is presented, particularly based on a spherical and a rod like structures. PMID:18815872

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

    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.

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

    Luciana Barros de Moura Neiva; Cassiane Dezoti da Fonseca; Mirian Watanabe; Maria de Fátima Fernandes Vattimo

    2013-01-01

    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/Bro...

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

    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).

  19. Nanophotonics and DNA: New approaches

    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...

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

    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.

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

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

    2003-01-01

    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......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...... 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...

  2. The ultrastructure of shelled and unshelled cashew nuts.

    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

  3. Mid-Infrared Spectroscopy of Polycyclic Aromatic Nitrogen Heterocycles (PANHS) and their Ions

    Mattioda, Andrew L.; Hudgin, Douglas; Bauschlicher, Charles W.; Alamandola, Louis J.

    2003-01-01

    In recent years, polycyclic aromatic nitrogen heterocycles (PANHs) have attracted a good deal of attention because of their potent carcinogenic and mutagenic properties, and their prevalence in our environment. Such species also play a prominent role in the chemistry of life up to and including the very nucleobases from which our DNA is constructed. Surprisingly, these compounds may even be common outside of our terrestrial environment. To wit, it is now widely accepted that polycyclic aromatic materials are abundant in space and represent a major reservoir of organic carbon in the interstellar medium and developing planetary systems. Given that nitrogen is the fourth most abundant chemically reactive element in space (surpassed only by hydrogen, carbon, and oxygen), it is entirely reasonable to suspect that PANHs may represent an important component of that organic reservoir. Motivated by their intrinsic merit and with special attention toward evaluating their exobiological significance, we have initiated a program to study the spectroscopic and chemical properties of P A " s under conditions relevant to extraterrestrial environments. Here we present the first results of that program-infrared spectroscopic measurements on a series of PANH"s in neutral and cationic forms, isolated in inert matrices at cryogenic temperatures.temperatures. The species studied include: 1 -, and 2-azabenz[a]anthracene, 1-, 2-, and 4- azachrysene, dibenz[a,h]acridine, and dibenz[a,J)acridine. The experimental measurements are also compared with theoretical spectra calculated using density functional theory. General spectroscopic trends observed in this series of compounds are discussed and the implications of these results for Astrophysics and Exobiology are considered.

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

    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

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

    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

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

    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

  7. Differential cytotoxic effects of arsenic compounds in human acute promyelocytic leukemia cells

    Arsenic trioxide, As2O3, has successfully been used to treat acute promyelocytic leukemia (APL). Induction of apoptosis in cancerous cells has been proposed to be the underlying mechanism for the therapeutic efficacy of arsenic. To further understand the cytotoxicity of arsenic compounds in APL cells, HL-60 cells were exposed to graded concentrations of the following arsenicals for up to 48 h: arsenic trioxide (AsIII), sodium arsenate (AsV), phenylarsine oxide (PAOIII), monomethylarsonous acid (MMAIII), monomethylarsonic acid (MMAV) and dimethylarsinic acid (DMAV), and the viability and modes of cell death assessed. The arsenic-exposed cells were stained with annexin V-PE and 7-aminoactinomycin D (7-AAD) and analyzed by flow cytometry in order to detect apoptotic and viable cells while cell morphology was visualized using scanning and transmission electron microscopy. Acridine orange staining and microtubule-associated protein 1 light chain 3 (MAP-LC3) detection were used to recognize autophagic cell death. The results showed that the compounds reduced viable HL-60 cells by inducing apoptosis in a concentration-dependent manner. None of the compounds tested caused a significant change in binding of acridine orange or redistribution of MAP-LC3. Potencies of the six different arsenic compounds tested were ranked as PAOIII > MMAIII ≥ AsIII > AsV > MMAV > DMAV. An increase in caspase-3 activity by PAOIII, MMAIII and DMAV implied that these compounds induced apoptosis in HL-60 cells through a caspase-dependent mechanism, but the other arsenic compounds failed to activate caspase-3, suggesting that they induce apoptosis by an alternative pathway.

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

    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

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

    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

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

    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

  11. 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

    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.

  12. Novel microfilaricidal activity of nanosilver

    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

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

    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.

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

    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

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

    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

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

    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

  17. Induction of cancer cell death by proton beam in tumor hypoxic region

    Proton beam has been applied to treat various tumor patients in clinical studies. However, it is still undefined whether proton radiation can inhibit the blood vessel formation and induce the cell death in vascular endothelial cells in growing organs. The aim of this study are first, to develop an optimal animal model for the observation of blood vessel development with low dose of proton beam and second, to investigate the effect of low dose proton beam on the inhibition of blood vessel formation induced by hypoxic conditions. In this study, flk1-GFP transgenic zebrafish embryos were used to directly visualize and determine the inhibition of blood vessels by low dose (1, 2, 5 Gy) of proton beam with spread out Bragg peak (SOBP). And we observed cell death by acridine orange staining at 96 hours post fertilization (hpf) stage of embryos after proton irradiation. We also compared the effects of proton beam with those of gamma-ray. An antioxidant, N-acetyl cystein (NAC) was used to investigate whether reactive oxygen species (ROS) were involved in the cell deaths induced by proton irradiation. Irradiated flk-1-GFP transgenic embryos with proton beam irradiation (35 MeV, spread out Bragg peak, SOBP) demonstrated a marked inhibition of embryonic growth and an altered fluorescent blood vessel development in the trunk region. When the cells with DNA damage in the irradiated zebrafish were stained with acridine orange, green fluorescent cell death spots were increased in trunk regions compared to non-irradiated control embryos. Proton beam also significantly increased the cell death rate in human umbilical vein endothelial cells (HUVEC), but pretreatment of N-acetyl cystein (NAC), an antioxidant, recovered the proton-induced cell death rate (p<0.01). Moreover, pretreatment of NAC abrogated the effect of proton beam on the inhibition of trunk vessel development and malformation of trunk truncation. From this study, we found that proton radiation therapy can inhibit the

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

    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.

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

    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.

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

    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

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

    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

  2. Cytotoxic Effects of Newly Synthesized Palladium(II Complexes of Diethyldithiocarbamate on Gastrointestinal Cancer Cell Lines

    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.

  3. Anti-tumor Effects of a Recombinant Fowlpox Virus Expressing Apoptin on Human Cervical Carcinoma in Vivo and in Vitro

    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.

  4. Allergy Testing and Drug Screening on an ITO-Coated Lab-on-a-Disc

    Ho Chin Kwok

    2016-02-01

    Full Text Available A lab-on-a-disc (LOAD is a centrifugal microfluidic set-up based on centrifugal force without using micro-pumps to drive reagents and cells to various chambers through channels and valves for reactions. A LOAD coated with conductive transparent indium tin oxide (ITO for thermal control was developed to screen allergy-blocking agents. When the acridine orange (AO-loaded KU-812 human basophilic cells were activated in the LOAD by stimuli, AO trapped in the cytoplasmic granules was released externally as an allergic mediator mimetic to report degranulation. This response was monitored by fluorescence when the released AO in supernatant had been transferred, with a higher spinning speed, from the reaction chamber to detection chamber in the LOAD where AO reacted with exogenous DNA. We report here the principles of the system and an improved LOAD set-up with the ITO-coated glass resistive microheater to run assays at 37 °C. By using this platform, we demonstrate here for the first time that triptolide, an active ingredient from the Chinese medicine herb Tripterygium wilfordii Hook f., was able to suppress the fMLP-mediated degranulation in basophils. This serves as an example how LOADs can be used to screen agents to alleviate symptoms of allergy.

  5. Composition, antimicrobial activity and in vitro cytotoxicity of essential oil from Cinnamomum zeylanicum Blume (Lauraceae).

    Unlu, Mehmet; Ergene, Emel; Unlu, Gulhan Vardar; Zeytinoglu, Hulya Sivas; Vural, Nilufer

    2010-11-01

    The essential oil from the bark of Cinnamomum zeylanicum Blume was analyzed by GC-MS and bioassays were carried out. Nine constituents representing 99.24% of the oil were identified by GC-MS. The major compounds in the oil were (E)-cinnamaldehyde (68.95%), benzaldehyde (9.94%) and (E)-cinnamyl acetate (7.44%). The antimicrobial activity of the oil was investigated in order to evaluate its efficacy against 21 bacteria and 4 Candida species, using disc diffusion and minimum inhibitory concentration methods. The essential oil showed strong antimicrobial activity against all microorganisms tested. The cytotoxic and apoptotic effects of the essential oil on ras active (5RP7) and normal (F2408) fibroblasts were examined by MTT assay and acridine orange/ethidium bromide staining, respectively. The cytotoxicity of the oil was quite strong with IC(50) values less than 20 μg/mL for both cell lines. 5RP7 cells were affected stronger than normal cells. Morphological observation of apoptotic cells indicated the induction of apoptosis at the high level of the oil, especially in 5RP7 cells. The present study showed the potential antimicrobial and anticarcinogenic properties of the essential oil of cinnamon bark, indicating the possibilities of its potential use in the formula of natural remedies for the topical treatment of infections and neoplasms. PMID:20828600

  6. Interaction of single and multi wall carbon nanotubes with the biological systems: tau protein and PC12 cells as targets.

    Zeinabad, Hojjat Alizadeh; Zarrabian, Alireza; Saboury, Ali Akbar; Alizadeh, Ali Mohammad; Falahati, Mojtaba

    2016-01-01

    Subtle changes in the structure of nanoparticles influence their surface tension and corresponding interaction with cells and proteins. Here, the interaction of the single wall carbon nanotube (SWCNT) and multiwall carbon nanotube (MWCNT) with different surface tension with tau protein was evaluated using a variety of techniques including far and near circular dichroism, fluorescence spectroscopy, dynamic light scattering, Zeta potential, and TEM evaluation. Also the cytotoxicity of SWCNT and MWCNT on the PC12 cell line as a model of nervous system cell line was investigated by the MTT, LDH, acridine orange/ethidium bromide staining, flow cytometry, caspase 3 activity, cell and membrane potential assays. It was observed that SWCNT induced more structural changes of tau protein relative to MWCNT/tau protein interaction. It was also revealed that SWCNT and MWCNT impaired the viability and complexity of PC12 cells in different modes of cytotoxicity. Analysis of cellular outcomes indicated that MWCNT in comparison with SWCNT resulted in induction of necrotic modes of cell death, whereas apoptotic modes of cell death were activated in SWCNT-incubated cells. Together these findings suggest that surface tension may be used to determine how nanoparticle structure affects neurotoxicity and protein conformational changes. PMID:27216374

  7. Transient absorption spectroscopy in biology using the Super-ACO storage ring FEL and the synchrotron radiation combination

    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,...

  8. Interaction of cationic dye/surfactants with Klebsiella K18 capsular polysaccharides: Physico-chemical studies

    Physico-chemical studies on the interaction of capsular polysaccharide (SPS) isolated from Klebsiella K18, with cationic dyes and surfactants have been reported. SPS is an integral component of gram-negative bacteria and having glucuronic acid as the potential anionic site, induced strong metachromasy (blue shift ∼ 110 nm) in the cationic dye pinacyanol chloride (PCYN). Reversal of metachromasy was observed upon addition of co-solvents which provides a qualitative measurement of stability and nature of metachromatic compound associated with PCYN-SPS interaction. Thermodynamic parameters such as association constant, changes in free energy, enthalpy and entropy of dye-polymer interaction, were evaluated which revealed the nature of interaction. Studies on fluorescence quenching of acridine orange (AO) was also performed. The interaction of SPS with cationic and cationic-non-ionic mixed surfactant systems have been studied by turbidimetry, spectrophotometry, spectrofluorometry and viscosity measurements. The studies could provide an understanding on the effects of the surfactants on binding with the polymer. The binding was found to be electrostatic in origin and also hydrophobic in nature to a certain extent.

  9. [Disinfectants - bacterial cells interactions in the view of hygiene and public health].

    Książczyk, Marta; Krzyżewska, Eva; Futoma-Kołoch, Bożena; Bugla-Płoskońska, Gabriela

    2015-01-01

    In recent years, the use of biocides has increased rapidly. One common example is triclosan, with wide application in households as well as medical and industrial fields, especially food industry and animal husbandry. Chemical disinfection is a major mean to control and eliminate pathogenic bacteria, particularly those with multidrug resistance (MDR) phenotype. However, exposition to biocides results in an adaptive response in microorganisms, causing them to display a wide range of resistance mechanisms. Numerous microorganisms are characterized by either natural resistance to chemical compounds or an ability to adapt to biocides using various strategies, such as: modification of cell surface structures (lipopolisaccharide), membrane fatty acids), over-expression of efflux pumps (a system for active transport of toxic compounds out of bacterial cell), enzymatic inactivation of biocides or altering biocide targets. For instance, it was shown that in vitro exposition of Salmonella Typhimurium to subinhibitory concentration of biocides (triclosan, quaternary ammonium compounds [QACs]) resulted in selection of variants resistant to tested biocides and, additionally, to acridine dyes and antibiotics. Bacillus subtilis and Micrococcus luteus strains isolated from chlorine dioxide containing disinfection devices were found to be resistant to chlorine dioxide and also to other oxidizing compounds, such as peracetic acid and hydrogen peroxide. Interaction between chemical compounds, including disinfectants and microbial cells, can create a serious threat to public health and sanitary-hygienic security. This phenomenon is connected with factor risk that intensify the probability of selection and dissemination of multidrug resistance among pathogenic bacteria. PMID:26400890

  10. Requirement of mitoses for the reversal of X-inactivation in cell hybrids between murine embryonal carcinoma cells and normal female thymocytes

    By means of a 5-bromodeoxyuridine (BrdU) incorporation and acridine orange fluorescence staining method the authors studied reactivation of the inactivated X chromosome (Xi) in newly formed cell hybrids between the near-diploid HPRT-deficient OTF9-63 murine embryonal carcinoma cell (ECC) with an XO sex chromosome constitution and the normal female mouse thymocyte. Synchronization of the late replicating S chromosome in such hybrid cells, indicative of reactivation, was found for the first time on Day 3, and the frequency of reactivation was attained 90% on Day 5. Inhibition of cell cycle progression either by methylglyoxal bis(guanylhydrazone) dihydrochloride, an inhibitor of polyamine metabolism, or by isoleucine-deficient medium after cell fusion delayed reactivation of the Xi, which implied that the number of cell division cycles traversed by individual cells rather than the length of time after cell fusion is critical for the reactivation. Double-labeling experiments using [3H]thymidine and BrdU indicated that hybrid cells had undergone three or four mitoses before reactivation of the Xi. Most probably reactivation of the Xi is consequent to reversion of the thymocyte genome to an undifferentiated state under the influence of OTF9 genome. DNA demethylation or dilution of Xi-specific factors by mitoses may be involved in this process