Novel photocatalyst-based colourimetric indicator for oxygen

The preparation and characterisation of a novel, UV-activated solvent-based, colourimetric indicator for O-2 is described, comprising a redox dye (methylene blue, MB), semiconductor photocatalyst (TiO2), and a sacrificial electron donor (SED), all dispersed/dissolved in a polymer medium (sulfonated polystyrene, SPS). Upon exposure, the indicator is readily photobleached as the MB is converted into its oxygen-sensitive, leuco form, LMB. Unlike its water-based counterpart, the recovery of the original colour is very slow (ca. 5 days cf. 6 min), probably due to the largely hydrophobic nature of the polymer encapsulation medium. The kinetics of film photobleaching appear to fit very well, in terms of: irradiance, [TiO2] and [MB], to the usual Langmuir-Hinshelwood type equation associated with a photocatalytic process. The glycerol appears not only to function as a SED, but also a plasticizer and medium for dye dissolution. The kinetics of colour recovery of the photobleached film appear directly dependent upon the ambient level of O-2 but shows a more complex dependence upon the relative humidity, RH. The photobleached film does not recover any of its colour over a 24 h period if the RH

Reversible, fluorescence-based optical sensor for hydrogen peroxide

The preparation and characterisation are described of a robust, reversible, hydrogen peroxide optical sensor, based on the fluorescent quenching of the dye ion-pair [Ru(bpy)(3)(2+)(Ph4B-)(2)], by O-2 produced by the catalytic breakdown of H2O2, utilizing the inorganic catalyst RuO2 center dot xH(2)O. The main feature of this system is the one-pot formulation of a coating ink that, when dried, forms an active single-layer fluorescence-based H2O2 sensor, demonstrably capable of detecting H2O2 over the range of 0.01 to 1 M, with a relative standard deviation of ca. 4% and a calculated lower limit of detection of 0.1 mM. These sensors are sterilisable, using dry-heat, and stable when stored over 40 days, without exhibiting any loss in sensitivity or response characteristics.

Photobleaching of methylene blue sensitised by TiO2: an ambiguous system?

The photobleaching of methylene blue MB, sensitised by TiO2, in an aqueous solution is studied in the absence and presence of oxygen, in the absence of oxygen and in the presence of a sacrificial electron acceptor (SED), MB is photoreduced to its colourless leuco form, LMB, by the TiO2 photocatalyst. This same photoreduction process is observed even if an SED is not present, indicating that MB itself can act as an SED. The oxidation of LMB by oxygen to regenerate MB is significantly slower if the aqueous solution is acidified (0.01 mol dm(-3) HClO4) and, at low partial pressures, the rate of reaction depends directly upon the concentration of dissolved oxygen. The TiO2-sensitised photobleaching of MB is irreversible in an oxygen-saturated aqueous solution, as expected, since the bleaching was due to an oxidative process. However, in an acidified solution (0.01 mol dm(-3) HClO4), the photobleaching process, in an oxygen-saturated solution, generates LMB initially. The latter situation arises because, under acidic conditions, LMB reacts only very slowly with oxygen to form MB. The significance of these findings with respect to the popular use of photobleaching of MB as a demonstration of semiconductor photomineralisation is discussed. (C) 1999 Elsevier Science S.A. All rights reserved.

Use of luminescent gold compounds in the design of thin-film oxygen sensors

The use of two gold compounds incorporated into thin plastic films as luminescence quenching oxygen sensors is described. The films are sensitive both to gaseous oxygen and to oxygen dissolved in nonaqueous media such as ethanol. The luminescence quenching of these sensors by oxygen obeys the Stern-Volmer equation and Stern-Volmer constants of 5.35 x 10(-3) and 0.9 x 10(-3) Torr(-1) are found, respectively, for the two dyes in a polystyrene polymer matrix. The sensitivity of the films is strongly influenced by the nature of the polymer matrix, and greatest sensitivity was found in systems based an the polymers polystyrene or cellulose acetate butyrate. Sensitivity was not found to be temperature dependent though raising the temperature hom 15 to 50 degrees C did result in a slight decrease in emission intensity and a hypsochromic shift in the emission wavelength. The rate of response and recovery of the sensors can be increased either by decreasing film thickness or by increasing the operating temperature. The operational and storage stability of these films is generally good though exposure to light should be avoided as one of the dyes tends to undergo photobleaching probably due to a photoinduced ligand substitution reaction.

MODELED DIFFUSION-CONTROLLED RESPONSE AND RECOVERY BEHAVIOR OF A NAKED OPTICAL FILM SENSOR WITH A HYPERBOLIC-TYPE RESPONSE TO ANALYTE CONCENTRATION

The diffusion-controlled response and recovery behaviour of a naked optical film sensor (i.e., with no protective membrane) with a hyperbolic-type response [i.e., S0/S = (1 + Kc), where S is the measured value of the absorbance or luminescence intensity of one form of the sensor dye in the presence of the analyte, S0 is the observed value of S in the absence of analyte and K is a constant] to changes in analyte concentration, c, in a system under test is approximated using a simple model, and described more accurately using a numerical model; in both models it is assumed that the system under test represents an infinite reservoir. Each model predicts the variations in the response and recovery times of such an optical sensor, as a function of the final external analyte concentration, the film thickness (I) and the analyte diffusion coefficient (D). From an observed signal versus time profile for a naked optical film sensor it is shown how values for K and D/I2 can be extracted using the numerical model. Both models provide a qualitative description of the often cited asymmetric nature of the response and recovery for hyperbolic-type response naked optical film sensors. It is envisaged that the models will help in the interpretation of the response and recovery behaviour exhibited by many naked optical film sensors and might be especially apposite when the analyte is a gas.

A specificity-enhanced time-resolved fluoroimmunoassay for zeranol employing the dry reagent all-in-one-well principle

A simple dry chemistry time-resolved fluorescence immunoassay (TR-FIA) method was developed for the measurement of zeranol in bovine urine samples. The samples were purified by immunoaffinity chromatography and a specificity-enhanced zeranol antibody was employed in the immunoassay. This resulted in a highly selective method, which had only negligible reactivity with Fusarium spp, toxins. The all-in-one-well dry chemistry concept made the assay very simple to use because all the assay-specific reagents were already present in the reaction wells in dry form. Only the addition of diluted sample extract was required to perform the competitive one-step TR-FIA and the results were available in less than 1 h. The analytical limit of detection (mean + 3s) for the immunoassay was 0.16 ng ml(-1) (n=12) and the functional limit of detection for the whole method, estimated by the analysis of zeranol-free samples, was 1.3 ng ml(-1) (n=20). The recovery of zeranol at the level of 2 ng ml(-1) was 99% (n=18) and the within-assay variation ranged between 4.5 and 9.0%.

DETERMINATION OF THE CONCENTRATIONS OF THE STEROIDS ESTRADIOL, PROGESTERONE AND TESTOSTERONE IN BOVINE SERA - COMPARISON OF COMMERCIAL DISSOCIATION ENHANCED LANTHANIDE FLUORESCENCE IMMUNOASSAY KITS WITH CONVENTIONAL RADIO AND ENZYME IMMUNOASSAYS

The performance of three conventional enzyme and radioimmunoassays routinely used to detect residues of anabolic steroids in cattle sera were compared with dissociation enhanced lanthanide fluorescence immunoassay (DELFIA) kits designed for the hospital market. Slight modifications to the kit reagents were required for the analysis of bovine sera. Owing to the large sample volumes used in conventional assays, detection limits were generally better than those obtained with DELFIA kits, however, assay reproducibility was enhanced using the DELFIA technology. Comparison of sera obtained from cattle implanted with anabolic steroids revealed a good correlation between alternate methods (r(2) from 0.91 to 0.97). The DELFIA kits offer a faster method for measuring estradiol, progesterone and testosterone with adequate sensitivity and in a safer environment than that encountered using radioimmunoassays.

Path-selective photoinduced electron transfer (PET) in a membrane-associated system studied by pH-dependent fluorescence

The pH-dependent fluorescence behavior of two regioisomeric 'receptor(1)-spacer(1)-fluorophore-spacer(2)-receptor(2)' systems 1 and 2 in micellar solutions of sodium dodecyl sulfate show that photoinduced electron transfer (PET) only occurs from the amine group connected to the 4-amino position of the aminonaphthalimide fluorophore in both cases. This demonstrates the directing influence of the photogenerated electric field within the aminonaphthalimide excited state on the electron transfer process. Since path-selectivity of PET is also known within the membrane-bound photosynthetic reaction center in bacteria, its origins may be illuminated by the simple experiments described here. (C) 2011 Elsevier B. V. All rights reserved.

Assessment of groundwater quality recovery strategies using nitrate transport modelling. Application to the Saône alluvial formations (Tournus, Saône-et-Loire)

To assess the efficiency of different agro-environmental strategies used to reduce groundwater pollution by nitrates, transport modelling in soils and groundwater has been carried out on two withdrawal areas in an alluvial plain. In a first time, the agro-environmental model AgriFlux allowed the simulation of water and nitrates fluxes flowing to groundwater. This model was calibrated for each agro-pedological unit of the studied territory. In a second time, the application of the hydrogeological model MODFLOW-MT3D allowed the simulation of nitrate transport in groundwater for the 1980-2004 period. This soil-groundwater coupled modelling has shown that soil nature is the first factor that conditions the vulnerability to nitrates. Thus, nitrate leaching occurs preferentially under sandy soils. Efficiency of different agro-environmental operations for groundwater quality recovery was quantified. The best results are obtained by combination of (1) grassland re-installation on sandy agricultural lots located in near well protection perimeter and (2) fertilization reduction on sandy agricultural lots located in the well alimentation area upstream the near protection perimeter. On other soils, the effect of grassland on groundwater quality improvement is more limited. Nevertheless, the control of nitrate fertilisation remains essential and is justified in both near and far well protection perimeters. Modelling thus allows optimising and priorizing agro-environmental actions in alluvial agricultural zones. [Comte J.-C., Banton O., Kockmann F., Villard A., Creuzot G. (2006), Assessment of groundwater quality recovery strategies using nitrate transport modelling. Application to the Saône alluvial formations (Tournus, Saône-et-Loire), Ingénieries Eau-Agriculture-Territoires, 45, 15-28]

The incidences of trisomy 8, trisomy 9 and D20S108 deletion in polycythaemia vera: an analysis of blood granulocytes using interphase fluorescence in situ hybridization:an analysis of blood granulocytes using interphase fluorescence in situ hybridization

We have used interphase fluorescence in situ hybridization (IFISH) to detect trisomy 8, trisomy 9 and 20q deletion in circulating granulocytes from patients with polycythaemia vera (PV). Out of 64 PV patients, 15 (23%) exhibited an abnormality. Two patients had trisomy 9, three had trisomy 8 and 10 patients had hemizygous deletion of D20S108 (a locus in the 20q common deleted region). Aberrant nuclei ranged from 10% to 80% in these 15 cases. There was no correlation between the presence of a marker and sex, age, interval between presentation and IFISH analysis, neutrophil or platelet count or therapy. Conventional marrow cytogenetic karyotype results were available in 23 cases and there was concurrence between these and blood IFISH in 16 cases (13 normal and three with 20q/D20S108 deletion by both methods). Three patients with D20S108 deletion by IFISH were normal by previous marrow cytogenetic testing and four cases with 20q deletion by previous marrow cytogenetics had normal blood granulocytes according to IFISH. Thus, we confirm that trisomies 8 and 9 and deletion of 20q are diagnostically useful markers of PV. IFISH analysis of blood granulocytes is a practical method for detecting these markers, but as an adjunct to, not as a substitute for, conventional marrow cytogenetics.

Gross anatomy of the muscle systems of Fasciola hepatica as visualized by phalloidin-fluorescence and confocal microscopy

Neuropeptides, biogenic amines and acetylcholine are expressed abundantly within the nervous systems of parasitic flatworms, and are particularly evident in the innervation of the musculature. Such associations have implicated the nervous system in locomotion, host attachment and reproductive co-ordination. Information on the muscle systems of parasitic flatworms is generally sparse, in particular those muscles associated with the reproductive system, intestinal tract and attachment apparatus. Also, the use of sectioned material has left description of the 3-dimensional organization of the musculature largely unrecorded. Using fluorescein isothiocyanate (FITC)-labelled phalloidin as a site-specific probe for filamentous actin, applied to whole-mount preparations of adult Fasciola hepatica and examined by confocal scanning laser microscopy, the present work reports on the organization of the major muscle systems in this trematode parasite. A highly regular array of outer circular, intermediate longitudinal and inner diagonal fibres distinguishes the body wall musculature, which is also involved in the development of both ventral and oral suckers. Circular fibres dominate the duct walls of the male and female reproductive systems, whereas the muscles of the intestinal tract have a somewhat diffuse arrangement of fibres. An understanding of the structural complexity of the muscle systems of parasitic flatworms is considered as fundamental to the interpretation of results from physiological experiments.