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Quencher
Kinds of Quencher Selected AbstractsHigh glucose activates pituitary proopiomelanocortin gene expression: possible role of free radical-sensitive transcription factorsDIABETES/METABOLISM: RESEARCH AND REVIEWS, Issue 4 2007Koichi Asaba Abstract Background Hyperglycemia is recognized as a metabolic stress, and indeed it is known to stimulate hypothalamo-pituitary-adrenal (HPA) axis, a representative anti-stress system, in patients with diabetes mellitus or in animal models of hyperglycemia. Thus, we tried to clarify the molecular mechanism of glucose-induced HPA axis activation. Methods We studied the effect of high glucose on the transcriptional regulation of proopiomelanocortin (POMC) gene that encodes adrenocorticotropic hormone, a central mediator of HPA axis, using AtT20 corticotroph cell line in vitro. Results We found that high glucose concentration (24 mM) significantly stimulated the 5,-promoter activity of POMC gene. The effect was promoter-specific, and was mimicked by nuclear factor-kappaB (NF-,B)- or AP1-responsive promoters but not by cAMP-responsive element or serum-response element-containing promoters. Furthermore, the stimulatory effect of high glucose on POMC gene was eliminated by NF-,B and AP1 inhibitors, suggesting the involvement of the transcriptional factors. The POMC 5,-promoter has the canonical NF-,B consensus sequence, and gel shift assay showed the binding of NF-,B to the element. Finally, the effect of high glucose was completely abolished by treatment with a radical quencher 4-hydroxy-2,2,6,6-tetramethylpiperidine-1-oxyl (TEMPOL). Conclusions Our data suggest that hyperglycemia activates POMC gene expression, at least partly, via NF-,B/AP1, and that high-glucose-induced free radical generation may mediate the activation of these transcription factors, which in turn stimulates the transcription of POMC gene. Copyright © 2006 John Wiley & Sons, Ltd. [source] A short proregion of trialysin, a pore-forming protein of Triatoma infestans salivary glands, controls activity by folding the N-terminal lytic motifFEBS JOURNAL, Issue 5 2008Rafael M. Martins Triatoma infestans (Hemiptera: Reduviidae) is a hematophagous insect that transmits the protozoan parasite Trypanosoma cruzi, the etiological agent of Chagas' disease. Its saliva contains trialysin, a protein that forms pores in membranes. Peptides based on the N-terminus of trialysin lyse cells and fold into ,-helical amphipathic segments resembling antimicrobial peptides. Using a specific antiserum against trialysin, we show here that trialysin is synthesized as a precursor that is less active than the protein released after saliva secretion. A synthetic peptide flanked by a fluorophore and a quencher including the acidic proregion and the lytic N-terminus of the protein is also less active against cells and liposomes, increasing activity upon proteolysis. Activation changes the peptide conformation as observed by fluorescence increase and CD spectroscopy. This mechanism of activation could provide a way to impair the toxic effects of trialysin inside the salivary glands, thus restricting damaging lytic activity to the bite site. [source] Conformational changes of ,-lactoglobulin in sodium bis(2-ethylhexyl) sulfosuccinate reverse micellesFEBS JOURNAL, Issue 4 2004A fluorescence, CD study The effect of ,-lactoglobulin encapsulation in sodium bis(2-ethylhexyl) sulfosuccinate reverse micelles on the environment of protein and on Trp was analysed at different water contents (,0). CD data underlined the distortion of the ,-sheet and a less constrained tertiary structure as the ,0 increased, in agreement with a concomitant red shift and a decrease in the signal intensity obtained in steady-state fluorescence measurements. Fluorescence lifetimes, evaluated by biexponential analysis, were ,1 = 1.28 ns and ,2 = 3.36 ns in neutral water. In reverse micelles, decay-associated spectra indicated the occurrence of important environmental changes associated with ,0. Bimolecular fluorescence quenching by CCl4 and acrylamide was employed to analyse alterations in the accessibility of the two Trp residues in ,-lactoglobulin, induced by changes in ,0. The average bimolecular quenching constant <> was found not to depend on ,0, confirming the insolubility of this quencher in the aqueous interface, while <> increases with ,0. The drastic decrease with ,0 of kq, associated with the longest lifetime, , comparatively to the increase of , emphasizes the location of ,-lactoglobulin in the aqueous interfacial region especially at ,0,,10. The fact that (,0 = 30) , (water) also confirms the important conformational changes of encapsulated ,-lactoglobulin. [source] Conjugated Polymers Combined with a Molecular Beacon for Label-Free and Self-Signal-Amplifying DNA MicroarraysADVANCED FUNCTIONAL MATERIALS, Issue 20 2009Kangwon Lee Abstract A conjugated polymer (CP) and molecular-beacon-based solid-state DNA sensing system is developed to achieve sensitive, label-free detection. A novel conjugated poly(oxadiazole) derivative exhibiting amine and thiol functional groups (POX-SH) is developed for unique chemical and photochemical stability and convenient solid-state on-chip DNA synthesis. POX-SH is soluble in most nonpolar organic solvents and exhibits intense blue fluorescence. POX-SH is covalently immobilized onto a maleimido-functionalized glass slide by means of its thiol group. Molecular beacons having a fluorescent dye or quencher molecule as the fluorescence resonance energy transfer (FRET) acceptor are synthesized on the immobilized POX-SH layer through direct on-chip oligonucleotide synthesis using the amine side chain of POX-SH. Selective hybridization of the molecular beacon probes with the target DNA sequence opens up the molecular beacon probes and affects the FRET between POX-SH and the dye or quencher, producing a sensitive and label-free fluorescence sensory signal. Various molecular design parameters, such as the size of the stem and loop of the molecular beacon, the choice of dye, and the number of quencher molecules are systematically controlled, and their effects on the sensitivity and selectivity are investigated. [source] State-state transitions for CCl2(X1A1, A1B1, a3B1) radical and collisional quenching of CCl2(A1B1 and a3B1) by O2, N2, NO, N2O, NH3, and various aminated moleculesINTERNATIONAL JOURNAL OF CHEMICAL KINETICS, Issue 6 2002Yide Gao CCl2 free radicals were produced by a pulsed dc discharge of CCl4 in Ar. Ground electronic state CCl2(X) radicals were electronically excited to the A1B1 (0,4,0) vibronic state with an Nd:YAG laser pumped dye laser at 541.52 nm. Experimental quenching data of excited CCl2(A1B1 and a3B1) by O2, N2, NO, N2O, NH3, NH(CH3)2, NH(C2H5)2, and N(C2H5)3 molecules were obtained by observing the time-resolved total fluorescence signal of the excited CCl2 radical in a cell, which showed a superposition of two exponential decay components under the presence of quencher. The quenching rate constants kA of CCl2(A) state and ka of CCl2(a) state were derived by analyzing the experimental data according to a proposed three-level model to deal with the CCl2(X1A1, A1B1, a3B1) system. The formation cross sections of complexes of electronically excited CCl2 radicals with O2, N2, NO, N2O, NH3, and aminated molecules were calculated by means of a collision-complex model. © 2002 Wiley Periodicals, Inc. Int J Chem Kinet 34: 351,356, 2002 [source] Kinetic Study of the Quenching Reaction of Singlet Oxygen by Common Synthetic Antioxidants (tert -Butylhydroxyanisol, tert -di-Butylhydroxytoluene, and tert -Butylhydroquinone) as Compared with ,-TocopherolJOURNAL OF FOOD SCIENCE, Issue 5 2009Ji In Kim ABSTRACT:, Effects of synthetic phenolic antioxidants (BHA, BHT, and TBHQ) on the methylene blue (MB) sensitized photooxidation of linoleic acid as compared with that of ,-tocopherol have been studied. Their antioxidative mechanism was studied by both ESR spectroscopy in a 2,2,6,6-tetramethylpiperidone (TMPD)-methylene blue (MB) system and spectroscopic analysis of rubrene oxidation induced by a chemical source of singlet oxygen. Total singlet oxygen quenching rate constants (kox,Q+kq) were determined using a steady state kinetic equation. TBHQ showed the strongest protective activity against the MB sensitized photooxidation of linoleic acid, followed by BHA and BHT. TBHQ (1 × 10,3 M) exhibited 86.5% and 71.4% inhibition of peroxide and conjugated diene formations, respectively, in linoleic acid photooxidation after 60-min light illumination. The protective activity of TBHQ against the photosensitized oxidation of linoleic acid was almost comparable to that of ,-tocopherol. The data obtained from ESR and rubrene oxidation studies clearly showed the strong singlet oxygen quenching ability of TBHQ. The kox,Q+kq of BHA, BHT, and TBHQ were determined to be 3.37 × 107, 4.26 × 106, and 1.67 × 108 M,1 s,1, respectively. The kox,Q+kq of TBHQ was within the same order of magnitude of that of ,-tocopherol, a known efficient singlet oxygen quencher. There was a high negative correlation (r2,=,,0.991) between log (kox,Q+kq) and reported oxidation potentials for the synthetic antioxidants, indicating their charge-transfer mechanism for singlet oxygen quenching. This is the 1st report on the kinetic study on kox,Q+kq of TBHQ in methanol as compared with other commonly used commercial synthetic antioxidants and ,-tocopherol. [source] Reconstitution of Photosystem II Reaction Center with Cu-Chlorophyll aJOURNAL OF INTEGRATIVE PLANT BIOLOGY, Issue 11 2006Shuang Liu Abstract An isolated photosystem (PS) II reaction center (RC) with altered pigment content was obtained by chemical exchange of native chlorophyll a (Chl) with externally added Cu-Chl a (Cu-Chl). Pigment composition and spectroscopic properties of the RC exchanged with Cu-Chl were compared with native RC and RC treated with Chl in the same way. High-performance liquid chromatography analysis showed approximately 0.5 Cu-Chl per two pheophytin in the Cu-Chl-reconstituted RC preparation. Insertion of Cu-Chl resulted in a decrease in absorption at 670 nm and an increase at 660 nm, suggesting that the peripheral Chl may have been displaced. Fluorescence emission spectra of the Cu-Chl-reconstituted RC displayed a marked decrease in fluorescence yield and a blue shift of the band maximum, accompanied by the appearance of a broad peak at a shorter wavelength, indicating that energy transfer in the modified RC was disturbed by Cu-Chl, a quencher of the excited state. However, there were few differences in the circular dichroism (CD) spectra, suggesting that the arrangement of pigments and proteins responsible for the CD signal was not significantly affected. In addition, no obvious change in peptide components was found after the exchange procedure. (Managing editor: Ping He) [source] Fluorescent, internally quenched, peptides for exploring the pH-dependent substrate specificity of cathepsin BJOURNAL OF PEPTIDE SCIENCE, Issue 7 2006Paolo Ruzza Abstract Cathepsin B is a cysteine protease that in tumor tissues is localized in both acidic lysosomes and extracellular spaces. It can catalyze the cleavage of peptide bonds by two mechanisms: endoproteolytic attack with a pH optimum around 7.4, and attack from the C -terminus with a pH optimum at 4.5,5.5. In this work, seven fluorescent, internally quenched, decapeptides have been synthesized using the prototypical cathepsin B selective substrate Z-Phe-Arg-AMC as a lead, and used to identify the structural factors determining the susceptibility of peptides to hydrolysis at acidic and neutral pH values. Each peptide differs from the others in one amino acid (residue 6) and contains a highly fluorescent Nma group linked to the ,-amino function of the N -terminal Orn residue and a Dnp group linked to the side chain of the Lys8 residue acting as a quencher. Proteolytic cleavage was monitored by measuring the increase of fluorescence at 440 nm upon excitation at 340 nm, and the cleavage sites were determined by HPLC followed by ESI-MS analysis. Peptides containing Ala or Phe at position 6 are good substrates for the enzyme at both pH 5.0 and 7.4. By contrast, those containing Glu, Asp, Lys or Val are not cleaved at all by cathepsin B at pH 7.4, and are poorly hydrolyzed at pH 5.0. These findings provide new information for the rational design of cathepsin B-activated peptide-containing anticancer drugs. Copyright © 2006 European Peptide Society and John Wiley & Sons, Ltd. [source] Melatonin generates singlet oxygen on laser irradiation but acts as a quencher when irradiated by lamp photolysisJOURNAL OF PINEAL RESEARCH, Issue 3 2005D. S. Maharaj Abstract:, Melatonin, a naturally occurring chemical mediator, although assigned a diverse range of functions, has attracted interest in recent years because of its ability to function as a free radical scavenger. Because of the implications of singlet oxygen in neurotoxicity, the objective of the study was to investigate the ability of melatonin to quench singlet oxygen generated using laser irradiation or lamp photolysis. The results show that melatonin produces radicals upon laser irradation while the lamp photolysis studies show that melatonin is able to scavenge singlet oxygen produced by naphthalene. While melatonin is a free radical scavenger under biological conditions, it acts as a generator of singlet oxygen and or radicals (as ,, is 1.41) when irradiated with laser light, implying that it has the potential to be used in photodynamic therapy in the destruction of tumors. [source] Scirpusin A, a hydroxystilbene dimer from Xinjiang wine grape, acts as an effective singlet oxygen quencher and DNA damage protectorJOURNAL OF THE SCIENCE OF FOOD AND AGRICULTURE, Issue 5 2010Qingjun Kong Abstract BACKGROUND: Grapes and red wines are rich sources of phenolic compounds such as anthocyanins, catechins, flavonols and stilbenes, most of which are potent antioxidants showing cardioprotective properties. We first isolated scirpusin A, a hydroxystilbene dimer, from a wine grape of Xinjiang, and studied its antioxidant activity. RESULTS: Reactive oxygen species scavenging effects and the protection against reactive singlet oxygen-induced DNA damage of scirpusin A have been investigated in our experiments. The concentration of scirpusin A required to inhibit 50% of 1O2 generation was 17 µmol L,1, while addition of scirpusin A at 140 µmol L,1 caused complete inhibition. Further kinetic study revealed that the reaction of Scirpusin A with singlet oxygen has an extremely high rate constant (ka = 4.68 × 109 L mol,1 s,1). Scirpusin A (140 µmol L,1) exhibited significant inhibition effects on pBR322 DNA breakage. However, scavenging effects of scirpusin A on superoxide anion O2,, and hydroxyl radical ·OH were not potent as the inhibitor rates at a concentration of 1400 µmol L,1 were 28.83% and 19.5%, respectively. CONCLUSION: The present study shows that scirpusin A is a selective quencher of singlet oxygen and a protector against reactive singlet oxygen-induced pBR322 DNA damage at very low concentrations. Copyright © 2010 Society of Chemical Industry [source] OT-674 Suppresses Photooxidative Processes Initiated by an RPE Lipofuscin FluorophorePHOTOCHEMISTRY & PHOTOBIOLOGY, Issue 1 2008Jilin Zhou The pathological processes involved in age-related macular degeneration (AMD) include retinal pigment epithelial (RPE) cell degeneration; oxidative mechanisms likely contribute to the demise of these cells. Indeed, RPE cells may be particularly susceptible to photooxidative mechanisms since they accumulate retinoid-derived photoreactive compounds that constitute the lipofuscin of the cell. Thus we undertook to test the capacity of OT-674, the reduction product (Tempol-H) of the nitroxide Tempol, to suppress photooxidative processes initiated by the RPE lipofuscin fluorophore A2E. Accordingly, when ARPE-19 cells that had accumulated A2E were irradiated at 430 nm, pretreatment with OT-674 (0.01,10 mm) was found to confer a resistance to cell death. Monitoring by quantitative HPLC also showed that OT-674 reduced A2E photooxidation in a cell-free system. Moreover, when presented with a singlet oxygen generator, OT-674 served as a quencher of singlet oxygen that was more effective than Trolox and ,-tocopherol. We conclude that OT-674 is a potent antioxidant that suppresses photooxidative processes generated in cultured RPE cells by the lipofuscin fluorophore A2E. As oxidative damage to RPE cells is considered to be a risk factor for AMD, antioxidant therapy with OT-674 may serve a protective role. [source] Dual Chromophore-Nitroxides: Novel Molecular Probes, Photochemical and Photophysical Models and Magnetic MaterialsPHOTOCHEMISTRY & PHOTOBIOLOGY, Issue 4 2007Gertz I. Likhtenstein Over the last decades scientists have faced growing requirements in novel methods of fast and sensitive analysis of antioxidant status of biological systems, spin redox probing and spin trapping, investigation of molecular dynamics, and of convenient models for studies of photophysical and photochemical processes. In approaching this problem, methods based upon the use of dual chromophore-nitroxide (CN) compounds have been suggested and developed. A CN consists of two molecular sub-functionality (a chromophore and a stable nitroxide radical) tethered together by spacers. In the dual compound the nitroxide is a strong intramolecular quencher of the fluorescence from the chromophore fragment. Reduction to hydroxylamine, oxidation of the nitroxide fragment or addition of an active radical yield the fluorescence increase and the parallel decay of the fragment electron spin resonance (ESR) signal. At certain conditions the dual molecules undergo photomagnetic switching and form excited state multi-spin systems. These unique properties of CN were intensively exploited as the basis for several methodologies, which include molecular probing, modeling intramolecular photochemical and photophysical processes, and construction of new magnetic materials. [source] Fusion,Fission Transport of Probes and Quenchers in Microdomains of an Amphiphilic Ionene Polyelectrolyte,PHOTOCHEMISTRY & PHOTOBIOLOGY, Issue 3 2007Celize M. Tcacenco In aqueous solution, amphiphilic ionenes such as the [3,22]-ionene spontaneously adopt globular conformations and form microdomains that are highly micelle-like, i.e. are capable of solubilizing organic molecules, binding and exchanging counterions and accelerating or inhibiting the rates of bimolecular reactions. Time-resolved fluorescence decay of pyrene and pyrene derivatives solubilized in these microdomains at concentrations where excimer formation occurs show that even water-insoluble probes can migrate between the hydrophobic microdomains formed in aqueous solution by a [3,22]-ionene chloride (with the N-terminal groups quaternized with benzyl chloride). Time-resolved studies of the quenching of pyrene fluorescence by alkylpyridine derivatives revealed similar behavior. The observed quenching behavior requires that the migration be between microdomains on the same ionene chain or same group of associated ionene chains and is consistent with migration dominated by fusion/fission transport of the probe and quencher. [source] The Effect of pH on the Topography of Porphyrins in Lipid Membranes,PHOTOCHEMISTRY & PHOTOBIOLOGY, Issue 2 2005Irena Bronshtein ABSTRACT The effect of the acidity of the environment on the topography and photophysics of sensitizer molecules in homogeneous solutions, and when embedded in a lipid microenvironment, was studied. Four hematoporphyrin (HP) analogs were studied, which have chemical "spacers" of varying lengths between the chromophoric tetrapyrrole and the carboxylate moiety. These derivatives have essentially the same chemical attributes and reactivity as the parent compound, HP IX, which is used in clinical procedures of photodynamic therapy. The binding constants of these HP derivatives to membrane model systems increase with the length of carboxylate chain in the pH range 3.0,6.6. This effect of chain length is attributed to an increase in the hydrophobicity of the molecule on elongation of the alkyl chains. A strong pH dependence of the quenching efficiency of the porphyrins' fluorescence by iodide ions was observed in aqueous solution and is attributed to a unique electrostatic interaction between the fluorophore and the quencher. The quenching efficiency in liposomes, relative to the quenching in buffer, as a function of pH, shows that porphyrins in the neutral form penetrate deeper inside the lipid bilayer and are less exposed to external quenching than when negatively charged at the carboxylic moiety. This vertical displacement in the membrane is also evidenced in the effect of pH on the photosensitized oxidation efficiency of a membrane-bound chemical target. Increasing the pH causes a significant decrease in the sensitization efficiency in liposomes. This trend is attributed to the vertical localization, and protonation of the carboxylic groups on lowering the pH leads to sinking of the sensitizer into the lipid bilayer and to a consequent generation of singlet oxygen at a deeper point. This increases the dwell time of singlet oxygen within the bilayer, which results in greater photodamage to a membrane-residing singlet oxygen target. [source] A Photophysical and Photochemical Study of 6-Methoxy-2-naphthylacetic Acid, the Major Metabolite of the Phototoxic Nonsteroidal Antiinflammatory Drug NabumetonePHOTOCHEMISTRY & PHOTOBIOLOGY, Issue 2 2000F. Boscá ABSTRACT Nabumetone is a phototoxic nonsteroidal antiinflammatory drug used for the treatment of osteoarthritis. However, nabumetone is considered a prodrug with its metabolite 6-methoxy-2-naphthylacetic acid the active form. Photophysical and photochemical studies on this metabolite have been undertaken. It undergoes photodecarboxylation in aerated aqueous and organic solvents. In addition to the accepted photodegradation pathway for related molecules, a new mechanism that implies generation of the naphthalene radical cation from the excited singlet and addition of O2 prior to the decarboxylation process has been demonstrated. Evidence for the involvement of the excited singlet state in this mechanism have been obtained by steady-state and time-resolved fluorescence experiments. The fluorescence quenching by O2 and the shorter singlet lifetime in aerated solvents support this assignment. Laser flash photolysis also supports this mechanism by showing the noninvolvement of the triplet in the formation of the naphthalene radical cation. Finally, the well-known electron acceptor CCl4 acts as an efficient singlet quencher, enhancing the route leading to the radical cation, preventing intersystem crossing to the triplet and thus resulting in a dramatic increase in the yield of 6-methoxy-2-naphthaldehyde, the major oxidative decarboxylation product; this constitutes unambiguous proof in favor of the new mechanistic proposals. [source] Controlling the photoluminescence of water-soluble conjugated poly[2-(3-thienyl)ethyloxy-4-butylsulfonate)] for biosensor applicationsPOLYMER INTERNATIONAL, Issue 5 2007Enrique López-Cabarcos Abstract The photoluminescence of poly[2-(3-thienyl)ethyloxy-4-butylsulfonate)] (PTE-BS) in aqueous solution increases threefold on addition of the surfactant tetrabutylammonium perchlorate (TBA). Furthermore, the luminescence of the PTE-BS/TBA system is reduced by more than five times by the addition of small amounts of the cationic electron acceptor methyl viologen (MV2+). The Stern,Volmer constant KSV = 1.4 × 104 L mol,1 for the quenching of the polymer,surfactant complex by MV2+ is approximately 60 times smaller than the KSV = 8.4 × 105 L mol,1 obtained in water polymer solutions without surfactant. Thus, the luminescence of PTE-BS in aqueous solution can be modulated by complexing the polymer either with a surfactant or with a quencher. In this contribution we show that the surfactant/quencher tuning effect found in polymers of the phenylenevinylene family, such as poly(2,5-methoxy-propyloxysulfonate phenylenevinylene), also appears in polymers of the thiophene family such as PTE-BS. Copyright © 2007 Society of Chemical Industry [source] False labelling due to quenching failure of N -hydroxy-succinimide,ester-coupled dyesPROTEINS: STRUCTURE, FUNCTION AND BIOINFORMATICS, Issue 7 2010Weiqun Wang Abstract In comparative fluorescence gel electrophoresis experiments, cross-talk was detected. It was traced back to a failure in the quenching process in typical labelling protocols. Despite a huge excess of potential reaction sites for the N -hydroxy-succinimide,ester-coupled dye, sufficient active dye molecules were available after the quenching step to label protein molecules un-specifically. It could be shown that only a 100-fold increase in the amount of quencher will silence residual dye to such an extent that no artificial signals are detected. [source] Simple PbII fluorescent probe based on PbII -catalyzed hydrolysis of phosphodiesterBIOPOLYMERS, Issue 6 2003Ming Sun Abstract A new fluorescent probe for PbII, p -nitrophenyl 3H -phenoxazin-3-one-7-yl phosphoric acid (NPPA), was designed and synthesized by linking resorufin (serving as a fluorophore and electron acceptor) to p -nitrophenol (serving as a fluorescence quencher and electron donor) through phosphodiester bonds. When NPPA was irradiated with light, intramolecular fluorescence self-quenching took place because of the photoinduced electron transfer from the donor to the acceptor. However, upon the addition of PbII, the phosphate ester bonds in the probe were cleaved and the fluorophore was released, accompanying the retrievement of fluorescence. © 2003 Wiley Periodicals, Inc. Biopolymers (Biospectroscopy), 2003 [source] An RNase H-Assisted Fluorescent Biosensor for AptamersCHEMBIOCHEM, Issue 12 2007Dae-Ro Ahn Dr. Recycling program. A signal amplification strategy was established for aptamer-based molecular recognition of thrombin with concomitant release of a single-stranded guard-DNA (g-DNA). The g-DNA then bound to F-RNA-Q, which contained a fluorophore and quencher. The fluorescence-quenched RNA was degraded by using RNase H to give a fluorescence signal, and the undamaged g-DNA was recycled to yield fluorescence amplification. [source] Catalysis and Rational Engineering of trans-Acting pH6DZ1, an RNA-Cleaving and Fluorescence-Signaling Deoxyribozyme with a Four-Way Junction StructureCHEMBIOCHEM, Issue 9 2006Yutu Shen Biosensor made of a catalytic DNA. When the RNA moiety (red) in the fluorogenic DNA substrate (purple) is cleaved by the bound catalytic DNA (blue) with a multiduplex structure, a strong fluorescence signal is generated owing to the separation of the fluorophore (F) from the quencher (Q). [source] Photoluminescence of Uranium(VI): Quenching Mechanism and Role of Uranium(V)CHEMISTRY - A EUROPEAN JOURNAL, Issue 27 2010Satoru Tsushima Dr. Abstract The photoluminescence of uranium(VI) is observed typically in the wavelength range 400,650,nm with the lifetime of several hundreds ,s and is known to be quenched in the presence of various halide ions (case,A) or alcohols (case,B). Here, we show by density functional theory (DFT) calculations that the quenching involves an intermediate triplet excited state that exhibits uranium(V) character. The DFT results are consistent with previous experimental findings suggesting the presence of photoexcited uranium(V) radical pair during the quenching process. In the ground state of uranyl(VI) halides, the ligand contributions to the highest occupied molecular orbitals increase with the atomic number (Z) of halide ion allowing larger ligand-to-metal charge transfer (LMCT) between uranium and the halide ion. Consequently, a larger quenching effect is expected as Z increases. The quenching mechanism is essentially the same in cases,A and B, and is driven by an electron transfer from the quencher to the UO22+ entity. The relative energetic stabilities of the triplet excited state define the "fate" of uranium, so that in case,A uranium(V) is oxidized back to uranium(VI), while in case,B uranium remains as pentavalent. [source] 2-Phenanthrenyl,DNA: Synthesis, Pairing, and Fluorescence PropertiesCHEMISTRY - A EUROPEAN JOURNAL, Issue 3 2009Nikolay Abstract Three 2,-phenanthrenyl- C -deoxyribonucleosides with donor (phenNH2), acceptor (phenNO2), or no (phenH) substitution on the phenanthrenyl core were synthesized and incorporated into oligodeoxyribonucleotides. Duplexes containing either one or three consecutive phenR residues, which were located opposite each other, were formed. Within these residues, the phenR residues are expected to recognize each other through interstrand stacking interactions, in much the same way as described previously for biphenyl DNA. The thermal, thermodynamic, and fluorescence properties of such duplexes were determined by UV melting analysis and fluorescence spectroscopy. Depending on the nature of the substituent, the thermal stability of single-modified duplexes can vary between ,2.7 to +11.3,°C in Tm and that of triple-modified duplexes from +7.8 to +11.1,°C. Van,t Hoff analysis suggested that the observed higher thermodynamic stability in phenH- and phenNO2 -containing duplexes is of enthalpic origin. A single phenH or phenNO2 residue in a bulge position also stabilizes a corresponding duplex. If a phenNO2 residue is placed in a bulge position next to a base mismatch this can lead, in a sequence-dependent manner, to duplex destabilization. The phenNO2 residue was found to be a highly efficient (10,100-fold) quencher of phenH and phenNH2 fluorescence if placed in the opposite position to the fluorophores. When phenH and phenNH2 residues were placed opposite each other, efficient quenching of phenH and enhancement of phenNH2 fluorescence was found, which is an indicator for electron- or energy-transfer processes between the aromatic units. [source] An Oligonucleotide-based Fluorescence Sensor for Mercury(II) in Aqueous SolutionsCHINESE JOURNAL OF CHEMISTRY, Issue 8 2009Huiwang WU Abstract A highly selective fluorescence sensor was developed for Hg(II) ion detection in aqueous solutions based on the selective binding of Hg(II) ions with a pair of thymine-thymine mismatch. The sensor consists of two DNA probes functionalized with a fluorophore (fluorescein, F) and a quencher (tetramethyl rhodamine, Q) moiety separately. This pair of DNA probes contains two pairs of thymine-thymine mismatches used to detect Hg(II) ions. In the presence of Hg(II) ions, thymine-Hg2+ -thymine was formed between thymine residues of probes. From that, the interaction of the two DNA probes increased. Thus, the DNA probes formed a double-stranded structure. Both the fluorophore and quencher were brought close to each other leading to the fluorescence resonance energy transfer (FRET) between F and Q. Under the optimum conditions, the sensor was used to detect the Hg(II) ions from 50 to 1000 nmol·L,1 with a regression equation y=5281.13,1650.56 lg[Hg2+] (R2=0.985). The linear range covers 100 to 500 nmol·L,1, and the limit of detection (LOD) is 79 nmol·L,1. The disturbance of some co-existing metal ions was explored, and no significant fluorescence quenching in the presence of 1.0 ,mol·L,1 other metal ions was observed. The fluorescence sensor has good sensitivity and selectivity for Hg(II) ions providing a rapid, simple and low cost method for the detection of mercury(II) ions in aqueous solutions. [source] Fluorescence quenching of anthracene by N, N -diethylaniline in the O/W microemulsionCHINESE JOURNAL OF CHEMISTRY, Issue 6 2000Xia Guo Abstract Photoinduced electron-transfer system of anthracene- N, N -diethylaniline (MA) was studied in the oil in water (O/W) microemulsions formed by SDS (sodium dodecyl sulfate), BA (benzyl alcohol) and H2O. The time-resolved fluorescence study showed mat the fluorescence quenching of the excited anthracene by DEA occurs at the interface of the O/W microemulsions. Besides as the quencher of the excited anthracene, N, N -diethylaniline could act as a cosurfactant to change the structures of the microemulsions, Just as BA did. The quenching rate constants for the different structures of the system were determined. [source] Highly selective single nucleotide polymorphism recogniton by a chiral (5S) PNA beacon,CHIRALITY, Issue 1 2009Filbert Totsingan Abstract A chiral peptide nucleic acid (PNA) beacon containing a C-5 modified monomer based on L-lysine was synthesized. The terminal amino group of the lysine side chain was linked to a spacer for future applications on surfaces. The PNA beacon bears a carboxyfluorescein fluorophore and a dabcyl quencher at opposite ends. The DNA binding properties were compared with those of a homologous PNA beacon containing only achiral monomers. Both beacons underwent a fluorescence increase in the presence of complementary DNA, with higher efficiency and higher selectivity (evaluated using single mismatched DNA sequences) observed for the chiral monomer containing PNA. Ion exchange (IE) HPLC with fluorimetric detection was used in combination with the beacon for the selective detection of complementary DNA. A fluorescent peak corresponding to the PNA beacon:DNA duplex was observed at a very low detection limit (1 nM). The discriminating capacity of the chiral PNA beacon for a single mismatch was found to be superior to those observed with the unmodified one, thus confirming the potency of chirality for increasing the affinity and specificity of DNA recognition. Chirality, 2009. © 2008 Wiley-Liss, Inc. [source] Anthraquinones as Artificial DNA Building BlocksEUROPEAN JOURNAL OF ORGANIC CHEMISTRY, Issue 13 2008Nicolas Bouquin Abstract Synthesis and properties of oligodeoxynucleotides containing anthraquinone-derived building blocks with flexible linkers are described. Starting from the 1,4-, 1,5-, 1,8- and 2,6-dihydroxyanthraquinone isomers, the corresponding phosphoramidites were prepared and incorporated into oligonucleotides. The site of linker attachment was found to be of critical importance for hybrid stability. Whereas the 2,6-isomer led to a significant stabilization, all other isomers had a negative effect on the stability of the duplex. Spectroscopic studies showed that the anthraquinones behave as fluorescence quenchers. Models of anthraquinone-modified double-stranded hybrids are proposed. (© Wiley-VCH Verlag GmbH & Co. KGaA, 69451 Weinheim, Germany, 2008) [source] Membrane embedded location of Na+ or H+ binding sites on the rotor ring of F1F0 ATP synthasesFEBS JOURNAL, Issue 22 2002Christoph Von Ballmoos Recent crosslinking studies indicated the localization of the coupling ion binding site in the Na+ -translocating F1F0 ATP synthase of Ilyobacter tartaricus within the hydrophobic part of the bilayer. Similarly, a membrane embedded H+ -binding site is accepted for the H+ -translocating F1F0 ATP synthase of Escherichia coli. For a more definite analysis, we performed parallax analysis of fluorescence quenching with ATP synthases from both I. tartaricus and E. coli. Both ATP synthases were specifically labelled at their c subunit sites with N -cyclohexyl- N, -(1-pyrenyl)carbodiimide, a fluorescent analogue of dicyclohexylcarbodiimide and the enzymes were reconstituted into proteoliposomes. Using either soluble quenchers or spinlabelled phospholipids, we observed a deeply membrane embedded binding site, which was quantitatively determined for I. tartaricus and E. coli to be 1.3 ± 2.4 Å and 1.8 ± 2.8 Å from the bilayer center apart, respectively. These data show a conserved topology among enzymes of different species. We further demonstrated the direct accessibility for Na+ ions to the binding sites in the reconstituted I. tartaricus c11 oligomer in the absence of any other subunits, pointing to intrinsic rotor channels. The common membrane embedded location of the binding site of ATP synthases suggest a common mechanism for ion transfer across the membrane. [source] A fluorescence quenching test for the detection of flavonoid transformationFEMS MICROBIOLOGY LETTERS, Issue 2 2001Lilian Schoefer Abstract A novel fluorescence quenching test for the detection of flavonoid degradation by microorganisms was developed. The test is based on the ability of the flavonoids to quench the fluorescence of 1,6-diphenyl-1,3,5-hexatriene (DPH). Several members of the anthocyanidins, flavones, isoflavones, flavonols, flavanones, dihydroflavanones, chalcones, dihydrochalcones and catechins were tested with regard to their quenching properties. The anthocyanidins were the most potent quenchers of DPH fluorescence, while the flavanones, dihydroflavanones and dihydrochalcones, quenched the fluorescence only weakly. The catechins had no visible impact on DPH fluorescence. The developed test allows a quick and easy differentiation between flavonoid-degrading and flavonoid-non-degrading bacteria. The investigation of individual reactions of flavonoid transformation with the developed test system is also possible. [source] A Smart Nanoprobe Based On Fluorescence-Quenching PEGylated Nanogels Containing Gold Nanoparticles for Monitoring the Response to Cancer TherapyADVANCED FUNCTIONAL MATERIALS, Issue 6 2009Motoi Oishi Abstract A biocompatible, caspase-3-responsive, and fluorescence-quenching smart apoptosis nanoprobe based on a PEGylated nanogel that contains gold nanoparticles (GNPs) (fluorescence quenchers) in the cross-linked polyamine gel core and fluorescein isothiocyanate (FITC)-labeled DEVD peptides at the tethered PEG chain ends is prepared for monitoring the cancer response to therapy. FITC,DEVD,nanogel,GNP shows very little fluorescence in the absence of activated caspase-3 (normal cells) through the fluorescence resonance energy transfer (FRET) process between the GNPs and the FITC molecules, while pronounced fluorescence signals are observed in apoptotic cells because of the cleavage of the DEVD peptide by activated caspase-3 present in the cells, which results in the release of FITC molecules. Thus, remarkable quenching and dequenching of fluorescence signals in response to activated caspase-3 is observed. Apoptotic cells are detected in human hepatocyte (HuH-7) multicellular tumor spheroids (MCTSs), a commonly used three-dimensional in vitro model mimicking the in vivo biology of tumors, as early as one day post-treatment with staurosporine, an apoptosis-inducing agent; while growth inhibition (i.e., change in size) of the HuH-7 MCTSs is only observed after a delay of three days (i.e., on day 4). This demonstrates the effectiveness of the FITC,DEVD,nanogel,GNP probe as a smart nanoprobe for real-time monitoring as well as a more rapid assessment of the early response to cancer therapy. [source] Functionalized Siloles: Versatile Synthesis, Aggregation-Induced Emission, and Sensory and Device ApplicationsADVANCED FUNCTIONAL MATERIALS, Issue 6 2009Zhen Li Abstract The synthesis of functionalized siloles has been a challenge because of the incompatibility of polar functional groups with the reactive intermediates in the conventional protocols for silole synthesis. In this work, a synthetic route for silole functionalization is elaborated, through which a series of functionalized siloles are successfully prepared. Whereas light emissions of traditional luminophores are often quenched by aggregation, most of the functionalized siloles show an exactly opposite phenomenon of aggregation-induced emission (AIE). The siloles are nonemissive when dissolved in their good solvents but become highly luminescent when aggregated in their poor solvents or in the solid state. Manipulation of the aggregation,deaggregation processes of the siloles enables them to play two seemly antagonistic roles and work as both excellent quenchers and efficient emitters. The AIE effect endows the siloles with multifaceted functionalities, including fluorescence quenching, pH sensing, explosive detection, and biological probing. The sensing processes are very sensitive (with detection limit down to 0.1,ppm) and highly selective (with capability of discriminating among different kinds of ions, explosives, proteins, DNAs, and RNAs). The siloles also serve as active layers in the fabrication of electroluminescent devices and as photosensitive films in the generation of fluorescence patterns. [source] | |||||||||||||||||||||||||||||||||||||