Home About us Contact | |||
Fluorescence Analysis (fluorescence + analysis)
Kinds of Fluorescence Analysis Selected AbstractsEvaluation of poly(ethylene-co-vinyl acetate-co-carbon monoxide) and polydimethylsiloxane for equilibrium sampling of polar organic contaminants in waterENVIRONMENTAL TOXICOLOGY & CHEMISTRY, Issue 9 2009Jörgen A Magnér Abstract Abstract-The aim of the present study was to develop a passive a bsorptive equilibrium sampler that would enable the determination of the concentrations of polar organic compound (POC) in water more efficiently than existing techniques. To this end, a novel plastic material, poly(ethylene-co-vinyl acetate-co-carbon monoxide) (PEVAC), was evaluated and the results were compared with an existing silicone-based passive absorptive equilibrium device. Seven compounds (imidacloprid, carbendazim, metoprolol, atrazin, carbamazepine, diazinon, and chlorpyrifos), a mixture of pharmaceuticals, and pesticides with a logarithmic octanol-water partition coefficient ranging from 0.2 to 4.77 were selected as model substances for the experiments. The results showed that six of the seven selected POCs reached distribution equilibrium within 4 d in the two materials tested. A linear relation with a regression coefficient of more than 0.8906 between the established logarithmic absorbent-water partition coefficient and the calculated logarithmic dissociation partition coefficient of the selected compounds in the two polymers was observed. The correlation between these two coefficients was within one order of magnitude for the compounds that reached equilibrium in the two polymers, which demonstrates that both materials are suitable for mimicking biological uptake of POCs. The PEVAC material showed an enhanced sorption for all selected compounds compared to the silicone material and up to five times higher enrichment for the most polar compound. Fluorescence analysis of the sampler cross-section, following the uptake of fluoranthene, and proof that the sorption was independent of surface area variations demonstrated that the PEVAC polymer possessed absorptive rather than adsorptive enrichment of organic compounds. [source] Major and trace element provenance signatures in stream sediments from the Kando River, San'in district, southwest JapanISLAND ARC, Issue 2 2006Edwin Ortiz Abstract Basement rocks in the catchment of the Kando River in southwest Japan can be divided into two main groups. Paleogene to Cretaceous felsic granitoids and volcanic rocks dominate in the upstream section, and more mafic, mostly Miocene volcanic and volcaniclastic rocks occur in the downstream reaches. Geochemically distinctive Mount Sambe adakitic volcanic products also crop out in the west. X-ray fluorescence analyses of major elements and 14 trace elements were made of two size fractions (<180 and 180,2000 µm) from 86 stream sediments collected within the catchment, to examine contrasts in composition between the fractions as a result of sorting and varying source lithotype. The <180 µm fractions are depleted in SiO2 and enriched in most other major and trace elements relative to the 180,2000 µm fractions. Na2O, K2O, Ba, Rb and Sr are either depleted relative to the 180,2000 µm fractions, or show little contrast in abundance. Sediments from granitoid-dominated catchments are distinguished by greater K2O, Th, Rb, Ba and Nb than those derived from the Miocene volcanic rocks. Granitoid-derived <180 µm fractions are also enriched in Zr, Ce and Y. Sediments derived from the Miocene volcanic rocks generally contain greater TiO2, Fe2O3*, Sc, V, MgO and P2O5, reflecting their more mafic source. Sediments containing Sambe volcanic rocks in their source are marked by higher Sr, CaO, Na2O and lower Y, reflecting an adakitic signature that persists into the lower main channel, where compositions become less variable as the bedload is homogenized. Normalization against source averages shows that compositions of the 180,2000 µm fractions are less fractionated from their parents than are the <180 µm fractions, which are enriched for some elements. Contrast between the size fractions is greatest for the granitoid-derived sediments. Weathering indices of the sediments are relatively low, indicating source weathering is moderate, and typical of temperate climates. Some zircon concentration has occurred in granitoid-derived <180 µm fractions relative to 180,2000 µm counterparts, but Th/Sc and Zr/Sc ratios overall closely reflect both provenance and homogenization in the lower reaches. [source] Capillary electrophoresis-laser induced fluorescence analysis of endogenous damage in mitochondrial and genomic DNAELECTROPHORESIS, Issue 13 2005Michaela Wirtz Abstract Reactive oxygen molecules are formed in vivo as by-products of normal aerobic metabolism. All organisms dependent on oxygen are inevitably exposed to these species so that DNA damage can occur in both genomic and mitochondrial DNA (mtDNA). In order to determine endogenous DNA damage we have developed an analytical method that involves the isolation and hydrolysis of genomic DNA or mtDNA, the labeling of modified and unmodified nucleotides and micellar electrokinetic chromatography with laser-induced fluorescence detection. With this method we have found etheno-adenine, thymine glycol, uracil, hypoxanthine, and 5-methylcytosine. These were identified by the addition of internal standards to the genomic or mtDNA. There are a large number of other signals in the electropherograms of mtDNA that we have never found in genomic DNA analysis because they are at lower concentration in the genome. In the DNA of untreated patients with chronic lymphocytic leukemia (CLL), uracil and high levels of etheno-adenine were found, which can be explained by antioxidant enzyme alterations and oxidative stress in the CLL lymphocytes. [source] Formation of Large Clusters in the Reaction of Lanthanide Cations with HeptamolybdateEUROPEAN JOURNAL OF INORGANIC CHEMISTRY, Issue 13 2004Katja Burgemeister Abstract A series of new lanthanide polyoxomolybdates was synthesised by reaction of lanthanide cations with the heptamolybdate anion (Mo7O24)6,. The heteropolyanions [Ln4(MoO4)(H2O)16(Mo7O24)4]14, (Ln = LaIII, CeIII, PrIII, SmIII, GdIII) were isolated as either their ammonium or tetramethylammonium salts and characterised by X-ray structure analysis, energy dispersive X-ray fluorescence analysis, and IR and Raman spectroscopy. The central [Ln4(MoO4)(H2O)16]10+ core of the anion is coordinated by four heptamolybdate groups. Each LnIII atom in the core exhibits ninefold coordination by oxygen atoms belonging to H2O, (MoO4)2, and (Mo7O24)6, units; the central molybdenum atom is coordinated tetrahedrally by four oxygen atoms. (© Wiley-VCH Verlag GmbH & Co. KGaA, 69451 Weinheim, Germany, 2004) [source] Lipid damage during frozen storage of Gadiform species captured in different seasonsEUROPEAN JOURNAL OF LIPID SCIENCE AND TECHNOLOGY, Issue 6 2007Santiago P. Aubourg Abstract Quality loss of two gadiform fish species (blue whiting, Micromesistius poutassou; hake, Merluccius merluccius) during frozen storage (,30 and ,10,°C; up to 12,months) was studied. For this, hydrolytic (formation of free fatty acids, FFA) and oxidative (conjugated dienes, peroxide and interaction compound formation) lipid damage were analysed. For both species, individual fishes captured in two different trials (May and November) were considered. Increasing (p,<0.05) lipid hydrolysis and oxidation (peroxide and interaction compound formation) were observed for all kinds of samples throughout the frozen storage. Interaction compound detection by fluorescence analysis showed the best correlation values with storage time. Some higher (p,<0.05) hydrolysis development could be observed in hake captured in May than in its counterpart from the November trial, while frozen blue whiting did not provide definite differences for FFA formation between both trials. Concerning peroxide formation, higher (p,<0.05) values were obtained for individual blue whiting and hake captured in November when compared to their corresponding May fish for both frozen storage conditions. Interaction compound formation was also found to be higher (p,<0.05) for November hake fish than for its counterpart captured in May, while blue whiting did not provide definite differences between trials. [source] Time-resolved fluorescence analysis of the recombinant photosystem II antenna complex CP29FEBS JOURNAL, Issue 2 2001Effects of zeaxanthin, phosphorylation Nonradiative dissipation of excitation energy is the major photoprotective mechanism in plants. The formation of zeaxanthin in the antenna of photosystem II has been shown to correlate with the onset of nonphotochemical quenching in vivo. We have used recombinant CP29 protein, over-expressed in Escherichia coli and refolded in vitro with purified pigments, to obtain a protein indistinguishable from the native complex extracted from thylakoids, binding either violaxanthin or zeaxanthin together with lutein. These recombinant proteins and the native CP29 were used to measure steady-state chlorophyll fluorescence emission and fluorescence decay kinetics. We found that the presence of zeaxanthin bound to CP29 induces a ,,35% decrease in fluorescence yield with respect to the control proteins (the native and zeaxanthin-free reconstituted proteins). Fluorescence decay kinetics showed that four components are always present but lifetimes (,) as well as relative fluorescence quantum yields (rfqy) of the two long-lived components (,3 and ,4) are modified by the presence of zeaxanthin. The most relevant changes are observed in the rfqy of ,3 and in the average lifetime (, 2.4 ns with zeaxanthin and 3.2,3.4 ns in the control proteins). When studied in vitro, no significant effect of acidic pH (5.2,5.3) is observed on chlorophyll a fluorescence yield or kinetics. The data presented show that recombinant CP29 is able to bind zeaxanthin and this protein-bound zeaxanthin induces a significant quenching effect. [source] Direct luminescence chronology of the Epipaleolithic Kebaran site of Nahal Hadera V, IsraelGEOARCHAEOLOGY: AN INTERNATIONAL JOURNAL, Issue 4 2003D.I. Godfrey-Smith We report direct luminescence ages for the culture-bearing sediments of the Kebaran site of Nahal Hadera V (NHV) in the coastal plain of Israel. Although the site contains, in addition to rich lithic deposits, plentiful mammalian bone, it has proved to be undatable using radiocarbon dating, in spite of the fact that the cultural context places the time of occupation well within the range of radiocarbon dating. In contrast, luminescence dating of the site sediments proved successful. Luminescence ages were determined using the single aliquot additive-dose (SAA) method, applied to sand-sized quartz extracts to determine past equivalent doses (De). Dose rates (R) were calculated using thick source alpha counting for the uranium (U) and thorium (Th) concentrations and x-ray fluorescence analysis for the potassium (K20) concentration. Of the five samples collected at the site, four represent cultural and subcultural deposits and the fifth represents the geological substrate for the archaeological deposit, a quartz-rich, carbonate-cemented dune sand known as aeolianite or kurkar. The luminescence age of the kurkar is 42.7 ± 6.3 ka. Human occupation of the site occurred between 21.3 ka and 14.0 ka ago, during the Last Glacial Maximum. © 2003 Wiley Periodicals, Inc. [source] Diffusion and distribution of element-labelled surfactants in human hairINTERNATIONAL JOURNAL OF COSMETIC SCIENCE, Issue 2 2004F.-J. Wortmann Synopsis To directly follow the diffusion process of cosmetically relevant agents into human hair, a specific methodological approach is presented and elucidated for selected surfactants. For this, practically relevant anionic and cationic surfactants were synthesized with a chlorine atom at the end of their alkyl chain. The property changes of the surfactants through the modification are corresponding to an extension of the alkyl chain by about two methylene groups, thus representing a moderate increase of hydrophobicity. After the application of a modified surfactant to hair, it can be localized and quantified through its chlorine atom in cross-sections by scanning electron microscopy combined with micro X-ray fluorescence analysis. The determination of the diffusion coefficient D is realized through the application of the Matano-equation to element intensity profiles. Values for D vary within the chosen range of pH and temperature between 10,14 and 10,16 m2 s,1. The diffusion coefficients for the anionic surfactants increase with decreasing pH and increasing temperature, The temperature dependence follows in all cases the Arrhenius relationship with activation energies EA of 50,100 kJ mol,1, which decrease with pH. The pH-related effects, with comparable values for D and EA, are opposite for the cationic surfactant. These observations are consistently interpreted on the basis of ionic and hydrophobic interactions in hair. Résumé Afin de suivre directement le processus de diffusion d'agents cosmétiques pertinents dans le cheveu humain, une approche méthodologique spécifique est présentée et approfondie pour des tensio actifs sélectionnés. En pratique, des tensio actifs pertinents, anioniques et cationiques ont été synthétisés en introduisant un atome de Chlore à l'extrémité de leur chaîne alkyle. Les changements de propriétés de ces tensio actifs, via cette modification, correspondent à un allongement de la chaîne alkyle d'environ deux groupes méthylène, représentant ainsi une augmentation modérée de leur caractère hydrophobe. Suite à l'application sur le cheveu d'un tensio actif modifié, il peut être localisé et quantifié par l'atome de Chlore dans des coupes transverses par microscopie électronique à balayage couplée à l'analyse par micro Fluorescence X. La détermination du coefficient de diffusion D est effectuée par l'application de l'équation de Matano aux profils de l'intensité de l'élément. Les valeurs de D varient, selon l'échelle de pH et de température, entre 10,14 et 10,16 m2 s,1. Les coefficients de diffusion pour les tensio actifs anioniques augmentent avec des pH décroissants et des températures croissantes. La dépendance vis à vis de la température suit, dans tous les cas, la relation d'Arrhenius avec des énergies d'activation EA de 50 à 100 kJ mol,1, qui décroît avec le pH. Les effets liés au pH, avec des valeurs comparables de D et EA, sont opposés pour le tensio actif cationique. Ces observations sont constamment interprétées par les interactions de types ioniques et hydrophobes dans le cheveu. [source] Effect of sequence distribution of PES/PEES random, block, and alternative copolymers on excimer formation in solutionJOURNAL OF APPLIED POLYMER SCIENCE, Issue 4 2007Xiao-Ming Zhou Abstract Three copolymers of poly(ether sulfone) and poly(ether ether sulfone) with the same composition but different sequence distribution were synthesized by three kinds of methods. Their molecular aggregation in dichloromethane was studied by fluorescence spectrophotometer and electron microscope. The experimental results revealed that the formation of intermolecular excimers in alternative copolymer (A50) dichloromethane solution were observed at a A50 concentration about 1.6 × 10,2 g/mL by the fluorescence analysis, but the formation of intermolecular excimers in dichloromethane were not found for random copolymer (R50) and block copolymer (B50). The electron micrograph of three copolymer films, heat-treated at 200°C for 7 days, presented a diffraction micrograph, which suggest that three copolymer molecular aggregation is changed from a randomly coiled amorphous phase to an ordered one, and the order structure of alternative copolymer (A50) was the most distinct in three copolymers. © 2007 Wiley Periodicals, Inc. J Appl Polym Sci 2007 [source] Bone material quality in transiliac bone biopsies of postmenopausal osteoporotic women after 3 years of strontium ranelate treatmentJOURNAL OF BONE AND MINERAL RESEARCH, Issue 4 2010Paul Roschger Abstract Strontium ranelate (SrR) is a relatively new treatment for osteoporosis. In this study we investigated its potential impact on human bone material quality in transiliac bone biopsies from postmenopausal osteoporotic women treated 3 years with calcium and vitamin D plus either 2,g SrR per day or placebo. Bone mineralization density distribution (BMDD), strontium (Sr) concentration, collagen cross-link ratio, and indentation modulus were analyzed by quantitative backscattered electron imaging, electron-induced X-ray fluorescence analysis, synchrotron radiation induced micro X-ray fluorescence elemental mapping, Fourier transform infrared imaging, and nanoindentation, respectively. The BMDD of SrR-treated patients was shifted to higher atomic numbers (Zmean +1.5%, p,<,.05 versus placebo). We observed Sr being preferentially incorporated in bone packets formed during SrR treatment up to 6% atom fraction [Sr/(Sr,+,Ca)] depending on the SrR serum levels of the individuals (correlation r,=,0.84, p,=,.018). Collagen cross-link ratio was preserved in SR-treated bone. The indentation modulus was significantly decreased in younger versus older bone packets for both placebo- (,20.5%, p,<,.0001) and SrR-treated individuals (,24.3%, p,<,.001), whereas no differences were found between the treatment groups. In conclusion, our findings indicate that after SrR treatment, Sr is heterogeneously distributed in bone and preferentially present in bone packets formed during treatment. The effect of SrR on BMDD seems to be due mainly to the uptake of Sr and not to changes in bone calcium content. Taken together, these data provide evidence that the investigated bone quality determinants at tissue level were preserved in postmenopausal osteoporotic women after 3-year treatment with 2,g SrR per day plus calcium and vitamin D. © 2010 American Society for Bone and Mineral Research [source] Determination of intrathyroidal iodine by X-ray fluorescence analysis in 60- to 65-year olds living in an iodine-sufficient areaJOURNAL OF INTERNAL MEDICINE, Issue 1 2006M. MILAKOVIC Abstract. Objectives., X-ray fluorescence (XRF) is a non-invasive method for determining the iodine content of the thyroid gland in vivo. In spite of the obvious clinical value of such a method in situations of iodine deficiency or iodine overload, the method has not so far been widely used. The objective was to investigate the applicability of the XRF method in a larger number of subjects. Design and subjects., The study comprised 37 individuals, aged 60,65 years, who had spent their entire life with iodine supplementation through iodinated table salt. Individuals with (previous) thyroid disease were excluded. The individual thyroid function had previously been evaluated by measurements of thyroid-related hormones, thyroid volume and 131-Iodine (131I) uptake which indicated a sufficient iodine intake of the population in the area. Iodine in the right thyroid lobe in each subject was examined using XRF. Results., The mean thyroid iodine concentration was 0.4 mg mL,1, corresponding to a mean total iodine content of 5.2 mg (range 0.9,20.2). There was a pronounced difference between individuals. No correlation was found between iodine concentration and 131I uptake or thyroid volume. Neither was iodine content and 131I uptake correlated. Conclusions., In a population living under iodine-sufficient conditions, a large variation of iodine stored in the thyroid is compatible with euthyroidism. Determination of the iodine pool by XRF investigation is feasible in a clinical setting and the method offers a unique possibility to study the intrathyroidal iodine pool in subjects with thyroid disease. The low radiation dose enables the use of the method in pregnant women and also in young individuals. [source] Adhesive bonding of super-elastic titanium,nickel alloy castings with a phosphate metal conditioner and an acrylic adhesiveJOURNAL OF ORAL REHABILITATION, Issue 6 2003H. Matsumura summary, The purpose of the current study was to evaluate the bonding characteristics of super-elastic titanium,nickel (Ti,Ni) alloy castings. Disk specimens were cast from a Ti,Ni alloy (Ti-50·85Ni mol%) using an arc centrifugal casting machine. High-purity titanium and nickel specimens were also prepared as experimental references. The specimens were air-abraded with alumina, and bonded with an adhesive resin (Super-Bond C & B). A metal conditioner containing a phosphate monomer (Cesead II Opaque Primer) was also used for priming the specimens. Post-thermocycling average bond strengths (MPa) of the primed groups were 41·5 for Ti,Ni, 30·4 for Ti and 19·5 for Ni, whereas those of the unprimed groups were 21·6 for Ti, 19·3 for Ti,Ni and 9·3 for Ni. Application of the phosphate conditioner elevated the bond strengths of all alloy/metals (P < 0·05). X-ray fluorescence analysis revealed that nickel was attached to the debonded resin surface of the resin-to-nickel bonded specimen, indicating that corrosion of high-purity nickel occurred at the resin,nickel interface. Durable bonding to super-elastic Ti,Ni alloy castings can be achieved with a combination of a phosphate metal conditioner and a tri- n -butylborane-initiated adhesive resin. [source] Ultraviolet curing of acrylic systems: Real-time Fourier transform infrared, mechanical, and fluorescence studiesJOURNAL OF POLYMER SCIENCE (IN TWO SECTIONS), Issue 23 2002Carmen Peinado Abstract The photopolymerization of acrylic-based adhesives has been studied by Fourier transform infrared and fluorescence analysis in real time. Real-time infrared spectroscopy reveals the influence of the nature of the photoinitiator on the kinetics of the reaction. Furthermore, the incident light intensity dependence of the polymerization rate shows that primary radical termination is the predominant mechanism during the initial stages of the curing of the acrylic system with bis(2,4,6-trimethylbenzoyl) phenyl phosphine oxide (TMBAPO) as a photoinitiator. The fluorescence intensity of selected probes increases during the ultraviolet curing of the adhesive, sensing microenvironmental viscosity changes. Depending on the nature of the photoinitiator, different fluorescence,conversion curves are observed. For TMBAPO, the fluorescence increases more slowly during the initial stage because of the delay in the gel effect induced by primary radical termination. Mechanical tests have been carried out to determine the shear modulus over the course of the acrylic adhesive ultraviolet curing. In an attempt to extend the applications of the fluorescence probe method, we have undertaken comparisons between the fluorescence changes and shear modulus. Similar features in both curves confirm the feasibility of the fluorescence method for providing information about microstructural changes during network formation. © 2002 Wiley Periodicals, Inc. J Polym Sci Part A: Polym Chem 40: 4236,4244, 2002 [source] The small-angle and wide-angle X-ray scattering set-up at beamline BL9 of DELTAJOURNAL OF SYNCHROTRON RADIATION, Issue 3 2007Christina Krywka The multi-purpose experimental endstation of beamline BL9 at the Dortmund Electron Accelerator (DELTA) is dedicated to diffraction experiments in grazing-incidence geometry, reflectivity and powder diffraction measurements. Moreover, fluorescence analysis and inelastic X-ray scattering experiments can be performed. Recently, a new set-up for small-angle and wide-angle X-ray scattering utilizing detection by means of an image-plate scanner was installed and is described in detail here. First small-angle X-ray scattering experiments on aqueous solutions of lysozyme with different cosolvents and of staphylococcal nuclease are discussed. The application of the set-up for texture analysis is emphasized and a study of the crystallographic texture of natural bio-nanocomposites, using lobster and crab cuticles as model materials, is presented. [source] Use of highly energetic (116,keV) synchrotron radiation for X-ray fluorescence analysis of trace rare-earth and heavy elementsJOURNAL OF SYNCHROTRON RADIATION, Issue 4 2001Izumi Nakai This study has revealed the advantages of the use of 116,keV X-rays as an excitation source of X-ray fluorescence (XRF) analyses. This technique is suitable for nondestructive multielemental analyses of heavy elements such as rare-earth elements. The lowest MDL value evaluated for the bulk analysis of a JG-1 standard reference sample (granite rock) was 0.1 p.p.m. for W for a 500,s measurement. The spectrum of standard glass samples of SRM612 demonstrated clearly resolved K -line peaks of more than 30 elements, including all the existing rare-earth elements, at 50 p.p.m. levels. The calibration curve for the determination of a rare-earth element shows a linear relation between the XRF intensity and concentrations from 10 to 0.03,ng. This powerful technique should be useful for nondestructive analyses of rare-earth and heavy elements in geological, geochemical and archaeological samples as well as industrial materials. [source] Clinical Pharmacokinetics of the PDT Photosensitizers Porfimer Sodium (Photofrin), 2-[1-Hexyloxyethyl]-2-Devinyl Pyropheophorbide-a (Photochlor) and 5-ALA-Induced Protoporphyrin IXLASERS IN SURGERY AND MEDICINE, Issue 5 2006David A. Bellnier PhD Abstract Background and Objectives Photodynamic therapy (PDT) uses a photosensitizer activated by light, in an oxygen-rich environment, to destroy malignant tumors. Clinical trials of PDT at Roswell Park Cancer Institute (RPCI) use the photosensitizers Photofrin, Photochlor, and 5-ALA-induced protoporphyrin IX (PpIX). In some studies the concentrations of photosensitizer in blood, and occasionally in tumor tissue, were obtained. Pharmacokinetic (PK) data from these individual studies were pooled and analyzed. This is the first published review to compare head-to-head the PK of Photofrin and Photochlor. Study Design/Materials and Methods Blood and tissue specimens were obtained from patients undergoing PDT at RPCI. Concentrations of Photofrin, Photochlor, and PpIX were measured using fluorescence analysis. A non-linear mixed effects modeling approach was used to analyze the PK data for Photochlor (up to 4 days post-infusion; two-compartment model) and a simpler multipatient-data-pooling approach was used to model PK data for both Photofrin and Photochlor (at least 150 days post-infusion; three-compartment models). Physiological parameters were standardized to correspond to a standard (70 kg; 1.818 m2 surface area) man to facilitate comparisons between Photofrin and Photochlor. Results Serum concentration-time profiles obtained for Photofrin and Photochlor showed long circulating half-lives, where both sensitizers could be found more than 3 months after intravenous infusion; however, estimated plasma clearances (standard man) were markedly smaller for Photofrin (25.8 ml/hour) than for Photochlor (84.2 ml/hour). Volumes of distribution of the central compartment (standard man) for both Photofrin and Photochlor were about the size (3.14 L, 4.29 L, respectively) of plasma volume, implying that both photosensitizers are almost 100% bound to serum components. Circulating levels of PpIX were generally quite low, falling below the level of instrument sensitivity within a few days after topical application of 5-ALA. Conclusion We have modeled the PK of Photochlor and Photofrin. PK parameter estimates may, in part, explain the relatively long skin photosensitivity attributed to Photofrin but not Photochlor. Due to the potential impact and limited experimental PK data in the PDT field further clinical studies of photosensitizer kinetics in tumor and normal tissues are warranted. Lasers Surg. Med. © 2006 Wiley-Liss, Inc. [source] Fluorescence spectroscopic analysis of circulating platelet activation during coronary angioplastyLASERS IN SURGERY AND MEDICINE, Issue 5 2001Alexander Christov PhD Abstract Background and Objective Platelet activation during percutaneous transluminal coronary angioplasty (PTCA) initiates thrombus formation and plaque regrowth at sites of arterial injury, limiting procedure efficacy. We have developed a simple assay for circulating platelet activation based on fluorescence analysis of membrane fluidity and intracellular calcium concentration and light scattering analysis of platelet aggregation. Study Design/Materials and Methods Platelet activation state was measured in 45 patients undergoing angioplasty, before and after treatment with platelet inhibitors. Results PTCA alone produced a decrease in pyrene dimer formation (P0.0083) and an increase in light scattering at 650 nm (P0.0128). Treatment with ADP and GPIIb/IIIa receptor antagonists reduced PTCA induced changes in pyrene dimer formation. An unexpected decrease in pyrene dimer formation (P0.05) was detected when the GPIIb/IIIa receptor antagonist was given together with an ADP receptor antagonist. Conclusions 1) Analysis of membrane fluidity provides a sensitive marker for platelet activation state. 2) Reduced membrane fluidity after combined platelet inhibitor treatments suggests reduced antiplatelet efficacy. Lasers Surg. Med. 39:414,426, 2001. © 2001 Wiley-Liss, Inc. [source] From cells to tissues: Fluorescence confocal microscopy in the study of histological samplesMICROSCOPY RESEARCH AND TECHNIQUE, Issue 2 2004Pietro Transidico Abstract Our knowledge of the genetic mechanisms controlling cell proliferation and differentiation usually originates from in vitro cultured cell line models. However, the definition of the molecular switches involved in control of homeostasis and the understanding of the changes occurring in neoplastic transformation require looking at single cells as the components of a complex tissue network. Histological examination of tissue samples can gain a substantial amount of information from high-resolution fluorescence analysis. In particular, confocal microscopy can help in the definition of functional pathways using multiparameter analysis. In this report, we present acquisition and analysis procedures to obtain high-resolution data from tissue sections. Confocal microscopy coupled to computational restoration, statistical evaluation of spatial correlations, and morphological analysis over large tissue areas were applied to colorectal samples providing a molecular fingerprint of the biological differences inferred from classical histological examination. Microsc. Res. Tech. 64:89,95, 2004. © 2004 Wiley-Liss, Inc. [source] Arabidopsis thaliana acyl-CoA-binding protein ACBP2 interacts with heavy-metal-binding farnesylated protein AtFP6NEW PHYTOLOGIST, Issue 1 2009Wei Gao Summary ,,Arabidopsis thaliana acyl-CoA-binding protein 2 (ACBP2) was observed to interact with farnesylated protein 6 (AtFP6), which has a metal-binding motif (M/LXCXXC). Their interaction and expression in response to heavy metals were investigated. ,,Yeast two-hybrid analysis and in vitro assays showed that an ACBP2 derivative lacking ankyrin repeats did not interact with AtFP6, indicating that the ankyrin repeats mediate protein,protein interaction. Autofluorescence-tagged ACBP2 and AtFP6 transiently co-expressed in tobacco (Nicotiana tabacum) were both targeted to the plasma membrane. ,,Reverse transcriptase polymerase chain reaction and northern blot analyses revealed that AtFP6 mRNA was induced by cadmium (Cd(II)) in A. thaliana roots. Assays using metal-chelate affinity chromatography demonstrated that in vitro translated ACBP2 and AtFP6 bound lead (Pb(II)), Cd(II) and copper (Cu(II)). Consistently, assays using fluorescence analysis confirmed that (His)6 -AtFP6 bound Pb(II), like (His)6 -ACBP2. ,,Arabidopsis thaliana plants overexpressing ACBP2 or AtFP6 were more tolerant to Cd(II) than wild-type plants. Plasma membrane-localized ACBP2 and AtFP6 probably mediate Pb(II), Cd(II) and Cu(II) transport in A. thaliana roots. Also, (His)6 -ACBP2 binds [14C]linoleoyl-CoA and [14C]linolenoyl-CoA, the precursors for phospholipid repair following lipid peroxidation under heavy metal stress at the plasma membrane. ACBP2 -overexpressing plants were more tolerant to hydrogen peroxide than wild-type plants, further supporting a role for ACBP2 in post-stress membrane repair. [source] The Effect of Decreasing Temperature up to Chilling Values on the in vivo F685/F735 Chlorophyll Fluorescence Ratio in Phaseolus vulgaris and Pisum sativum: The Role of the Photosystem I Contribution to the 735 nm Fluorescence Band ,PHOTOCHEMISTRY & PHOTOBIOLOGY, Issue 1 2000Giovanni Agati ABSTRACT The effect of leaf temperature (T), between 23 and 4°C, on the chlorophyll (Chl) fluorescence spectral shape was investigated under moderate (200 ,E m,2 s,1) and low (30,35 ,E m,2 s,1) light intensities in Phaseolus vulgaris and Pisum sativum. With decreasing temperature, an increase in the fluorescence yield at both 685 and 735 nm was observed. A marked change occurred at the longer emission band resulting in a decrease in the Chl fluorescence ratio, F685/F735, with reducing T. Our fluorescence analysis suggests that this effect is due to a temperature-induced state 1,state 2 transition that decreases and increases photosystem II (PSII) and photosystem I (PSI) fluorescence, respectively. Time-resolved fluorescence lifetime measurements support this interpretation. At a critical temperature (about 6°C) and low light intensity a sudden decrease in fluorescence intensity was observed, with a larger effect at 685 than at 735 nm. This is probably linked to a modification of the thylakoid membranes, induced by chilling temperatures, which can alter the spillover from PSII to PSI. The contribution of photosystem I to the long-wavelength Chl fluorescence band (735 nm) at room temperature was estimated by both time-resolved fluorescence lifetime and fluorescence yield measurements at 685 and 735 nm. We found that PSI contributes to the 735 nm fluorescence for about 40, 10 and 35% at the minimal (F0), maximal (Fm) and steady-state (Fs) levels, respectively. Therefore, PSI must be taken into account in the analysis of Chl fluorescence parameters that include the 735 nm band and to interpret the changes in the Chl fluorescence ratio that can be induced by different agents. [source] Mutations of genes in synthesis of the carotenoid precursors of ABA lead to pre-harvest sprouting and photo-oxidation in riceTHE PLANT JOURNAL, Issue 2 2008Jun Fang Summary Pre-harvest sprouting (PHS) or vivipary in cereals is an important agronomic trait that results in significant economic loss. A considerable number of mutations that cause PHS have been identified in several species. However, relatively few viviparous mutants in rice (Oryza sativa L.) have been reported. To explore the mechanism of PHS in rice, we carried out an extensive genetic screening and identified 12 PHS mutants (phs). Based on their phenotypes, these phs mutants were classified into three groups. Here we characterize in detail one of these groups, which contains mutations in genes encoding major enzymes of the carotenoid biosynthesis pathway, including phytoene desaturase (OsPDS), ,-carotene desaturase (OsZDS), carotenoid isomerase (OsCRTISO) and lycopene , -cyclase (,-OsLCY), which are essential for the biosynthesis of carotenoid precursors of ABA. As expected, the amount of ABA was reduced in all four phs mutants compared with that in the wild type. Chlorophyll fluorescence analysis revealed the occurrence of photoinhibition in the photosystem and decreased capacity for eliminating excess energy by thermal dissipation. The greatly increased activities of reactive oxygen species (ROS) scavenging enzymes, and reduced photosystem (PS) II core proteins CP43, CP47 and D1 in leaves of the Oscrtiso/phs3-1mutant and OsLCY RNAi transgenic rice indicated that photo-oxidative damage occurred in PS II, consistent with the accumulation of ROS in these plants. These results suggest that the impairment of carotenoid biosynthesis causes photo-oxidation and ABA-deficiency phenotypes, of which the latter is a major factor controlling the PHS trait in rice. [source] PAUL-LOUIS CYFFLÉ'S (1724,1806) TERRE DE LORRAINE: A TECHNOLOGICAL STUDYARCHAEOMETRY, Issue 5 2010M. MAGGETTI Fragments of four Terre de Lorraine biscuit figurines were subjected to porosity analysis, X-ray fluorescence analysis, X-ray diffraction analysis, backscattered-electron image analysis,coupled with energy dispersive spectrometry,and electron backscatter diffraction analysis to determine the porosity, bulk, major, minor and trace element compositions, and the composition and the proportion of their constituent phases. Cyfflé's Terre de Lorraine wares embrace two distinct types of paste, a calcareous and an aluminous,siliceous one. Both are porous (9,25% water adsorption). The former consists of a mixture of different proportions of ground quartz or calcined flint, ground Pb-bearing glass and calcium carbonate with a refractory clay. The firing temperature was between 950 and 1050°C. For the latter, Cyfflé mixed ground pure amorphous SiO2, ground quartz or calcined flint, ground porcelain, ground Na,Ca-glass and coarse-grained kaolinite with a fine-grained kaolinitic clay. The figurines were fired below 1000°C. The result was a porous, hard paste porcelain-like material. Cyfflé's recipes for both pastes can be calculated from the chemical and the modal analyses. [source] THE HISTORY OF THE LAWRENCE BERKELEY NATIONAL LABORATORY INSTRUMENTAL NEUTRON ACTIVATION ANALYSIS PROGRAMME FOR ARCHAEOLOGICAL AND GEOLOGICAL MATERIALSARCHAEOMETRY, Issue 2 2007F. ASARO The Lawrence Berkeley National Laboratory pottery provenance group developed standards and instrumental neutron activation analysis (INAA) methods that are used at many archaeometry laboratories around the world. The background and development of ,Standard Pottery' and of methods for INAA are described. Early pottery provenance studies are described, and other research programmes, involving obsidian and magmatic mixing, the origin of the stone used for the Colossi of Memnon, and the ,Plate of Brass', are mentioned. Research work by the Laboratory included the discovery of the world-wide iridium anomaly and extensive subsequent research on what has come to be known as the ,Asteroid Impact Theory'. Characteristics of the analytical programme for pottery provenance work, including overall aims, precision and accuracy, intercalibration, and irradiation and measurement protocols, are discussed. New research areas developed in the past 15 years, to broaden the usefulness of chemical compositional data for archaeological investigation, and examples of recent work, are described. This research, which makes use of high-precision X-ray fluorescence analysis in addition to INAA measurements on sample splits, includes distinguishing the products of different workshops located at the same production site, studies on the significance of the distribution of silver in archaeological pottery and the use of high-precision chemical compositional data as an aid for making chronological distinctions. [source] Wavelength-dispersive X-ray fluorescence analysis of ancient glasses,ARCHAEOMETRY, Issue 4 2002R. Falcone The application of X-ray fluorescence to the study of the composition of ancient glasses has been limited by the large amount of material required for samples. In this paper the setting of a reduced sampling XRF method for the analysis of ancient glasses is described. The method involves the preparation of glass beads by melting the sample with lithium tetraborate flux. By means of reference and synthetic samples, the method allows regression curves to be set, covering broad ranges from ppm to high concentrations. By means of a single program, up to 31 elements of interest in the study of ancient glass are analysed. The reproducibility, sensitivity and reliability of the method are discussed. The results demonstrate that 0.2 g of glass is sufficient to obtain accurate and sensitive analyses for most of the elements of interest. [source] Screening of chondrogenic factors with a real-time fluorescence-monitoring cell line ATDC5-C2ER: Identification of sorting nexin 19 as a novel factorARTHRITIS & RHEUMATISM, Issue 11 2009Akinori Kan Objective To establish a cell culture system for noninvasive and real-time monitoring of chondrogenic differentiation in order to screen for chondrogenic factors. Methods The optimum reporter construct transfected into chondrogenic ATDC5 cells was selected by a luciferase reporter assay and fluorescence analysis during cultures with insulin. The established cell line was validated according to its fluorescence following stimulation with SOX proteins, bone morphogenetic protein 2 (BMP-2), or transforming growth factor , (TGF,) and was compared with the level of messenger RNA for COL2A1 as well as with the degree of Alcian blue staining. Screening of chondrogenic factors was performed by expression cloning using a retroviral expression library prepared from human tracheal cartilage. The expression pattern of the identified molecule was examined by in situ hybridization and immunohistochemistry. Functional analysis was performed by transfection of the identified gene, the small interfering RNA, and the mutated gene. Results We established an ATDC5 cell line with 4 repeats of a highly conserved enhancer ligated to a COL2A1 basal promoter and the DsRed2 reporter (ATDC5-C2ER). Fluorescence was induced under the stimulations with SOX proteins, BMP-2, or TGF,, showing good correspondence to the chondrogenic markers. Screening using the ATDC5-C2ER system identified several chondrogenic factors, including sorting nexin 19 (SNX19). SNX19 was expressed in the limb cartilage of mouse embryos and in the degraded cartilage of adult mouse knee joints during osteoarthritis progression. The gain-of-function and loss-of-function analyses revealed a potent chondrogenic activity of SNX19. Conclusion We established the ATDC5-C2ER system for efficient monitoring of chondrogenic differentiation by fluorescence analysis, and we identified a novel chondrogenic factor (SNX19) using this system. This system will be useful for elucidating the molecular network of chondrogenic differentiation. [source] Cloning, expression, purification and crystallization as well as X-ray fluorescence and preliminary X-ray diffraction analyses of human ADP-ribosylhydrolase 1ACTA CRYSTALLOGRAPHICA SECTION F (ELECTRONIC), Issue 5 2009Stefan Kernstock Human ADP-ribosylhydrolase 1 (hARH1, ADPRH) cleaves the glycosidic bond of ADP-ribose attached to an Arg residue of a protein. hARH1 has been cloned, expressed heterologously in Escherichia coli, purified and crystallized in complex with K+ and ADP. The orthorhombic crystals contained one monomer per asymmetric unit, exhibited a solvent content of 43% and diffracted X-rays to a resolution of 1.9,Å. A prerequisite for obtaining well diffracting crystals was the performance of X-ray fluorescence analysis on poorly diffracting apo hARH1 crystals, which revealed the presence of trace amounts of K+ in the crystal. Adding K-ADP to the crystallization cocktail then resulted in a crystal of different morphology and with dramatically improved diffraction properties. [source] Characterization of the G-quadruplex structure of a catalytic DNA with peroxidase activityBIOPOLYMERS, Issue 5 2009De-Ming Kong Abstract It has been reported that the complexes formed by hemin and some G-quadruplexes can be developed as a new class of DNAzyme with peroxidase activity. This kind of DNAzyme has received a great deal of attention. But to date, the actual G-quadruplex structure that can provide hemin with enhanced peroxidase activity is in doubt. Herein, the G-quadruplex structure of CatG4, a 21-nucleotide DNA oligomer which was previously reported to bind hemin and the resulting complex exhibiting enhanced peroxidase activity, was characterized by fluorescence and circular dichroism measurements. The results suggest that the catalytically active form of CatG4 may be a unimolecular parallel quadruplex rather than a unimolecular chair-type antiparallel quadruplex or a multistranded parallel quadruplex. In addition, the fluorescence analysis of labeled oligonucleotides may be developed as a supplementary tool for the study of DNA conformations. © 2009 Wiley Periodicals, Inc. Biopolymers 91: 331,339, 2009. This article was originally published online as an accepted preprint. The "Published Online" date corresponds to the preprint version. You can request a copy of the preprint by emailing the Biopolymers editorial office at biopolymers@wiley.com [source] The ybeY protein from Escherichia coli is a metalloproteinACTA CRYSTALLOGRAPHICA SECTION F (ELECTRONIC), Issue 11 2005Chenyang Zhan The three-dimensional crystallographic structure of the ybeY protein from Escherichia coli (SwissProt entry P77385) is reported at 2.7,Å resolution. YbeY is a hypothetical protein that belongs to the UPF0054 family. The structure reveals that the protein binds a metal ion in a tetrahedral geometry. Three coordination sites are provided by histidine residues, while the fourth might be a water molecule that is not seen in the diffraction map because of its relatively low resolution. X-ray fluorescence analysis of the purified protein suggests that the metal is a nickel ion. The structure of ybeY and its sequence similarity to a number of predicted metal-dependent hydrolases provides a functional assignment for this protein family. The figures and tables of this paper were prepared using semi-automated tools, termed the Autopublish server, developed by the New York Structural GenomiX Research Consortium, with the goal of facilitating the rapid publication of crystallographic structures that emanate from worldwide Structural Genomics efforts, including the NIH-funded Protein Structure Initiative. [source] |