Sol gel based fiber optic sensor for blook pH measurement

International Nuclear Information System (INIS)

Grant, S. A.; Glass, R. S.

1996-01-01

This paper describes a fiber-optic pH sensor based upon sol-gel encapsulation of a self-referencing dye, seminaphthorhodamine-1 carboxylate (SNARF-1C). The simple sol-gel fabrication procedure and low coating leachability are ideal for encapsulation and immobilization of dye molecules onto the end of an optical fiber. A miniature bench-top fluorimeter system was developed for use with the optical fiber to obtain pH measurements. Linear and reproducible responses were obtained in human blood in the pH range 6.8 to 8.0, which encompasses the clinically-relevant range. Therefore, this sensor can be considered for in vivo use

Cytotoxicity Test Based on Human Cells Labeled with Fluorescent Proteins: Fluorimetry, Photography, and Scanning for High-Throughput Assay.

Science.gov (United States)

Kalinina, Marina A; Skvortsov, Dmitry A; Rubtsova, Maria P; Komarova, Ekaterina S; Dontsova, Olga A

2018-06-01

High- and medium-throughput assays are now routine methods for drug screening and toxicology investigations on mammalian cells. However, a simple and cost-effective analysis of cytotoxicity that can be carried out with commonly used laboratory equipment is still required. The developed cytotoxicity assays are based on human cell lines stably expressing eGFP, tdTomato, mCherry, or Katushka2S fluorescent proteins. Red fluorescent proteins exhibit a higher signal-to-noise ratio, due to less interference by medium autofluorescence, in comparison to green fluorescent protein. Measurements have been performed on a fluorescence scanner, a plate fluorimeter, and a camera photodocumentation system. For a 96-well plate assay, the sensitivity per well and the measurement duration were 250 cells and 15 min for the scanner, 500 cells and 2 min for the plate fluorimeter, and 1000 cells and less than 1 min for the camera detection. These sensitivities are similar to commonly used MTT (tetrazolium dye) assays. The used scanner and the camera had not been previously applied for cytotoxicity evaluation. An image processing scheme for the high-resolution scanner is proposed that significantly diminishes the number of control wells, even for a library containing fluorescent substances. The suggested cytotoxicity assay has been verified by measurements of the cytotoxicity of several well-known cytotoxic drugs and further applied to test a set of novel bacteriotoxic compounds in a medium-throughput format. The fluorescent signal of living cells is detected without disturbing them and adding any reagents, thus allowing to investigate time-dependent cytotoxicity effects on the same sample of cells. A fast, simple and cost-effective assay is suggested for cytotoxicity evaluation based on mammalian cells expressing fluorescent proteins and commonly used laboratory equipment.

Assessment of uranium in ground water from Durg District of Chhattisgarh state and its correlation with other quality parameters

International Nuclear Information System (INIS)

Santosh Kumar Sar; Megha Sahu; Shweta Singh; Vijita Diwan; Manoj Jindal; Arun Arora

2017-01-01

The contamination of uranium in the ground water samples gathered from different source of Durg District (Chhattisgarh) have been analysed by using LED Fluorimeter. It was observed that uranium concentration ranges from 0.638 µg/l to 45.7 µg/l and 100% of the samples found under the safe limit of 60 μg/l prescribed by the India’s Atomic Energy Regulatory Board, Department of Atomic Energy, 2004. In present work, an attempt has been made to inspect uranium concentration in the samples and its correlation with the physico-chemical properties of water. A positive correlation of uranium with TDS, conductance, chloride, nitrate, sulphate, hardness, calcium, magnesium and alkalinity was found with various water quality parameters. (author)

Determination of uranium in environmental samples from the Nile delta and the adjacent regions of Egypt using laser fluorimetry

Energy Technology Data Exchange (ETDEWEB)

Shawky, S; Ibrahim, N; Farouk, A [National research Center for Nuclear safety and radiation Control, Atomic Energy Authority, cairo (Egypt); Ghods, A [IAEA Laboratories, Chemistry Unit., A 244 Seibersdorf, (Austria)

1995-10-01

Total uranium content was determined in soil and plant samples taken from various areas in the Nile delta. Samples from east and west of the delta, the suez canal` cities and from alexandria region were analyzed using laser fluorimetry (LF). Uranium was extracted from digested ashed samples with methyl-isobutyl ketone (MIBK) and measured using laser fluorimeter. The radium content of the same soil samples was determined using gamma spectrometry (GS). The uranium content of plant samples was determined using (LF), since it has a detection limit lower than of (GS). The uranium content varied between 0.6-4.4{mu}g/g and 0.032-0.17 {mu}g/g for soil and plant samples respectively. 1 fig., 3 tabs.

Determination of uranium in environmental samples from the Nile delta and the adjacent regions of Egypt using laser fluorimetry

International Nuclear Information System (INIS)

Shawky, S.; Ibrahim, N.; Farouk, A.; Ghods, A.

1995-01-01

Total uranium content was determined in soil and plant samples taken from various areas in the Nile delta. Samples from east and west of the delta, the suez canal' cities and from alexandria region were analyzed using laser fluorimetry (LF). Uranium was extracted from digested ashed samples with methyl-isobutyl ketone (MIBK) and measured using laser fluorimeter. The radium content of the same soil samples was determined using gamma spectrometry (GS). The uranium content of plant samples was determined using (LF), since it has a detection limit lower than of (GS). The uranium content varied between 0.6-4.4μg/g and 0.032-0.17 μg/g for soil and plant samples respectively. 1 fig., 3 tabs

A specific alpha laboratory dedicated to structural and thermodynamic studies on actinide complexes

Energy Technology Data Exchange (ETDEWEB)

Charbonnel, M-C.; Berthon, C.; Berthon, L.; Boubals, N.; Couston, L.; Auwer, C. den; Meyer, D.; Guillaneux, D

2004-07-01

The main scope of the LN1 laboratory in ATALANTE facility is the chemical and physico-chemical study of transuranic samples to understand the behavior of compounds of actinide with selective ligands at a molecular scale. The main techniques implemented in this laboratory are the following ones: Nuclear Magnetic Resonance spectrometer (400 MHz shielded magnetic field), a four circle X-ray diffractometer for single crystals, a microcalorimeter to the measurement of low heats of reactions, a Time Resolved Laser-induced spectro-fluorimeter, vibrational spectrometers: FTIR and Raman, an Electro-spray Ionisation Mass spectrometer. Specific glove boxes have been built for each technique to work on radio elements with safety conditions and allow the analysis of samples in different states (aqueous and organic liquids, gels, solids). (authors)

Laser fluorimetric determination of uranium in environmental samples from Nile Delta and adjacent regions

International Nuclear Information System (INIS)

Shawky, S.; Ibrahiem, N.; Farouk, A.; Ghods, A.

1994-01-01

Total uranium content was determined in soil and plant samples obtained from various areas in the Nile Delta. Samples taken from east and west of the delta, Suez canal cities and from the Alexandria region were analysed using laser fluorimetry (LF). Uranium was extracted from digested samples with methyl-isobutyl ketone and measured using a laser fluorimeter. The radium content of the same soil samples was determined using gamma spectroscopy. The uranium content of plant samples was determined using LF, since this technique has a detection limit lower than that of GS. Uranium content in the samples varied between 0.6-4.4 μg/g for soil and 0.032-0.17 μg/g for plant tissue. (author)

Estimation of uranium in bioassay samples of occupational workers by laser fluorimetry

International Nuclear Information System (INIS)

Suja, A.; Prabhu, S.P.; Sawant, P.D.; Sarkar, P.K.; Tiwari, A.K.; Sharma, R.

2010-01-01

A newly established uranium processing facility has been commissioned at BARC, Trombay. Monitoring of occupational workers at regulars intervals is essential to assess intake of uranium by the workers in this facility. The design and engineering safety features of the plant are such that there is very low probability of uranium getting air borne during normal operations. However, the leakages from the system during routine maintenance of the plant may result in intake of uranium by workers. As per the new biokinetic model for uranium, 63% of uranium entering the blood stream gets directly excreted in urine. Therefore, bioassay monitoring (urinalysis) was recommended for these workers. A group of 21 workers was selected for bioassay monitoring to assess the existing urinary excretion levels of uranium before the commencement of actual work. For this purpose, sample collection kit along with an instruction slip was provided to the workers. Bioassay samples received were wet ashed with conc. nitric acid and hydrogen peroxide to break down the metabolized complexes of uranium and it was co-precipitated with calcium phosphate. Separation of uranium from the matrix was done using ion exchange technique and final activity quantification in these samples was done using laser fluorimeter (Quantalase, Model No. NFL/02). Calibration of the laser fluorimeter is done using 10 ppb uranium standard (WHO, France Ref. No. 180000). Verification of the system performance is done by measuring concentration of uranium in the standards (1 ppb to 100 ppb). Standard addition method was followed for estimation of uranium concentration in the samples. Uranyl ions present in the sample get excited by pulsed nitrogen laser at 337.1 nm, and on de-excitation emit fluorescence light (540 nm) intensity which is measured by the PMT. To estimate the uranium in the bioassay samples, a known aliquot of the sample was mixed with 5% sodium pyrophosphate and fluorescence intensity was measured

Ratiometric, filter-free optical sensor based on a complementary metal oxide semiconductor buried double junction photodiode.

Science.gov (United States)

Yung, Ka Yi; Zhan, Zhiyong; Titus, Albert H; Baker, Gary A; Bright, Frank V

2015-07-16

We report a complementary metal oxide semiconductor integrated circuit (CMOS IC) with a buried double junction (BDJ) photodiode that (i) provides a real-time output signal that is related to the intensity ratio at two emission wavelengths and (ii) simultaneously eliminates the need for an optical filter to block Rayleigh scatter. We demonstrate the BDJ platform performance for gaseous NH3 and aqueous pH detection. We also compare the BDJ performance to parallel results obtained by using a slew scanned fluorimeter (SSF). The BDJ results are functionally equivalent to the SSF results without the need for any wavelength filtering or monochromators and the BDJ platform is not prone to errors associated with source intensity fluctuations or sensor signal drift. Copyright © 2015 Elsevier B.V. All rights reserved.

Use of zero order diffraction of a grating monochromator towards convenient and sensitive detection of fluorescent analytes in multi fluorophoric systems

Science.gov (United States)

Panigrahi, Suraj Kumar; Mishra, Ashok Kumar

2018-02-01

White light excitation fluorescence (WLEF) is known to possess analytical advantage in terms of enhanced sensitivity and facile capture of the entire fluorescence spectral signature of multi component fluorescence systems. Using the zero order diffraction of the grating monochromator on the excitation side of a commercial spectrofluorimeter, it has been shown that WLEF spectral measurements can be conveniently carried out. Taking analyte multi-fluorophoric systems like (i) drugs and vitamins spiked in urine sample, (ii) adulteration of extra virgin olive oil with olive pomace oil and (iii) mixture of fabric dyes, it was observed that there is a significant enhancement of measurement sensitivity. The total fluorescence spectral response could be conveniently analysed using PLS2 regression. This work brings out the ease of the use of a conventional fluorimeter for WLEF measurements.

LUMOS - A Sensitive and Reliable Optode System for Measuring Dissolved Oxygen in the Nanomolar Range

DEFF Research Database (Denmark)

Lehner, Philipp; Larndorfer, Christoph; Garcia-Robledo, Emilio

2015-01-01

Most commercially available optical oxygen sensors target the measuring range of 300 to 2 mu mol L-1. However these are not suitable for investigating the nanomolar range which is relevant for many important environmental situations. We therefore developed a miniaturized phase fluorimeter based...... for read out of less sensitive optical oxygen sensors based on the same or similar indicator dyes, for example for monitoring oxygen at physiological conditions. The presented sensor system exhibits lower noise, higher resolution and higher sensitivity than the electrochemical STOX sensor previously used...... measurement system called the LUMOS (Luminescence Measuring Oxygen Sensor). It consists of a readout device and specialized "sensing chemistry" that relies on commercially available components. The sensor material is based on palladium(II)-5,10,15,20-tetrakis-(2,3,4,5,6-pentafluorphenyl)-porphyrin embedded...

226Ra and natural uranium in egyptian bottled mineral waters

International Nuclear Information System (INIS)

Higgy, R.H.

2000-01-01

Concentration levels of 226 Ra and natural uranium have been analysed bottled mineral water commercially available in egypt. 226 Ra was determined by applying a chemical procedure in which Ra was coprecipitated with Ba as sulphate. The precipitate was then dissolved with EDTA and then measured by liquid scintillation system, after mixing with a scintillation cocktail. Natural uranium was determined by applying a chemical procedure for uranium extraction using MIBK and then measured using laser fluorimeter system. The concentration values obtained were compared with concentrations reported by other countries and with reference values accepted for drinking water. Based on the consumption rate and the measured concentrations, the collective committed effective doses were calculated. In addition, Ca, Mg and Na were measured using Icp system and compared with some worldwide values

Gating circuit for single photon-counting fluorescence lifetime instruments using high repetition pulsed light sources

International Nuclear Information System (INIS)

Laws, W.R.; Potter, D.W.; Sutherland, J.C.

1984-01-01

We have constructed a circuit that permits conventional timing electronics to be used in single photon-counting fluorimeters with high repetition rate excitation sources (synchrotrons and mode-locked lasers). Most commercial time-to-amplitude and time-to-digital converters introduce errors when processing very short time intervals and when subjected to high-frequency signals. This circuit reduces the frequency of signals representing the pulsed light source (stops) to the rate of detected fluorescence events (starts). Precise timing between the start/stop pair is accomplished by using the second stop pulse after a start pulse. Important features of our design are that the circuit is insensitive to the simultaneous occurrence of start and stop signals and that the reduction in the stop frequency allows the start/stop time interval to be placed in linear regions of the response functions of commercial timing electronics

Quality Assurance Project Plan For Verification of ANDalyze Lead100 Test Kit and AND1000 Fluorimeter

Science.gov (United States)

Lead (Pb) is a naturally occurring metal in the aquatic environment; however, most Pb contamination of concern arises from anthropogenic sources (such as deposition of Pb dust from combustion processes in natural waterways or due to its use in plumbing materials). Although Pb is...

A Fluorescence Based Miniaturized Detection Module for Toxin Producing Algae

Science.gov (United States)

Zieger, S. E.; Mistlberger, G.; Troi, L.; Lang, A.; Holly, C.; Klimant, I.

2016-12-01

Algal blooms are sensitive to external environmental conditions and may pose a serious threat to marine and human life having an adverse effect on the ecosystem. Harmful algal blooms can produce different toxins, which can lead to massive fish kills or to human disorders. Facing these problems, miniaturized and low-cost instrumentation for an early detection and identification of harmful algae classes has become more important over the last years. 1,2Based on the characteristic pigment pattern of different algae classes, we developed a miniaturized detection module, which is able to detect and identify algae classes after analyzing their spectral behavior. Our device combines features of a flow-cytometer and fluorimeter and is build up as a miniaturized and low-cost device of modular design. Similar to a fluorimeter, it excites cells in the capillary with up to 8 different excitation wavelengths recording the emitted fluorescence at 4 different emission channels. Furthermore, the device operates in a flow-through mode similar to a flow-cytometer, however, using only low-cost elements such as LEDs and photodiodes. Due to its miniaturized design, the sensitivity and selectivity increase, whereas background effects are reduced. With a sampling frequency of 140 Hz, we try to detect and count particular cell events even at a concentration of 2 cells / 7.3 µL illuminated volume. Using a self-learning multivariate algorithm, the data are evaluated autonomously on the device enabling an in-situ analysis. The flexibility in choosing excitation and emission wavelengths as well as the high sampling rate enables laboratory applications such as measuring induction kinetics. However, in its first application, the device is part of an open and modular monitoring system enabling the sensing of chemical compounds such as toxic and essential Hg, Cd, Pb, As and Cu trace metal species, nutrients and species related to the carbon cycle, VOCs and potentially toxic algae classes (FP7

A new trend to determine biochemical parameters by quantitative FRET assays.

Science.gov (United States)

Liao, Jia-yu; Song, Yang; Liu, Yan

2015-12-01

Förster resonance energy transfer (FRET) has been widely used in biological and biomedical research because it can determine molecule or particle interactions within a range of 1-10 nm. The sensitivity and efficiency of FRET strongly depend on the distance between the FRET donor and acceptor. Historically, FRET assays have been used to quantitatively deduce molecular distances. However, another major potential application of the FRET assay has not been fully exploited, that is, the use of FRET signals to quantitatively describe molecular interactive events. In this review, we discuss the use of quantitative FRET assays for the determination of biochemical parameters, such as the protein interaction dissociation constant (K(d)), enzymatic velocity (k(cat)) and K(m). We also describe fluorescent microscopy-based quantitative FRET assays for protein interaction affinity determination in cells as well as fluorimeter-based quantitative FRET assays for protein interaction and enzymatic parameter determination in solution.

Regions with increased natural radioactivity. Pt. 4

International Nuclear Information System (INIS)

Sansoni, B.; Matthes, W.; Schnitzler, J.

1986-05-01

Between May 23 and June 23, 1984, 169 specimens were taken of surface water and drinking water in the Fichtelgebirge, mainly from natural springs and wells in the granite region as well as in part from the drinking water supply. The uranium concentration was measured in the measuring van by a laser fluorimeter UA-3 (Scintrex); the water temperature, electrical conductivity, oxygen concentration and the pH value were determined by the Combibox (WTW). Furthermore, the discharge from the springs and wells was also determined. The concentration of radon-222 was emanometrically determined the following day in the field laboratory, while the radium-226 was separated overnight from the 20-l water specimens by ion exchange at manganese dioxide columns. The radium-226 was later determined emanometrically from the eluate in Juelich. Multi-element determinations by ICP atomic emission, atomic fluorescence and atomic absorption spectrometry were carried out in selected water specimens. (orig./HP) [de

A new front-face optical cell for measuring weak fluorescent emissions with time resolution in the picosecond time scale.

Science.gov (United States)

Gryczynski, Z; Bucci, E

1993-11-01

Recent developments of ultrafast fluorimeters allow measuring time-resolved fluorescence on the picosecond time scale. This implies one is able to monitor lifetimes and anisotropy decays of highly quenched systems and of systems that contain fluorophores having lifetimes in the subnanosecond range; both systems that emit weak signals. The combination of weak signals and very short lifetimes makes the measurements prone to distortions which are negligible in standard fluorescence experiments. To cope with these difficulties, we have designed a new optical cell for front-face optics which offers to the excitation beam a horizontal free liquid surface in the absence of interactions with optical windows. The new cell has been tested with probes of known lifetimes and anisotropies. It proved very useful in detecting tryptophan fluorescence in hemoglobin. If only diluted samples are available, which cannot be used in front-face optics, regular square geometry can still be utilized by inserting light absorbers into a cuvette of 1 cm path length.

Development of fluorescent, oscillometric and photometric methods to determine absorbed dose in irradiated fruits and nuts

International Nuclear Information System (INIS)

Kovacs, A.; Foeldiak, G.; Hargittai, P.; Miller, S.D.

2001-01-01

To ensure suitable quality control at food irradiation technologies and for quarantine authorities, simple routine dosimetry methods are needed for absorbed dose control. Taking into account the requirements at quarantine locations these methods would require nondestructive analysis for repeated measurements. Different dosimetry systems with different analytical evaluation methods have been tested and/or developed for absorbed dose measurements in the dose range of 0.1-10 kGy. In order to use the well accepted ethanolmonochlorobenzene dosimeter solution and the recently developed aqueous alanine solution in small volume sealed vials, a new portable, digital, and programmable oscillometric reader was developed. To make use of the availability of the very sensitive fluorimetric evaluation method, liquid and solid inorganic and organic dosimetry systems were developed for dose control using a new routine, portable, and computer controlled fluorimeter. Absorption or transmission photometric methods were also applied for dose measurements of solid or liquid phase dosimeter systems containing radiochromic dye agents, which change colour upon irradiation. (author)

A Quantitative Fluorescence-Based Lipase Assay

Directory of Open Access Journals (Sweden)

Giovanna Lomolino

2012-01-01

Full Text Available An easy and fast gel diffusion assay for detecting and monitoring lipase activity by quantification of fluorescein is described. By measuring the intensity of fluorescein, it is possible to obtain a calibration curve with a regression coefficient better than by using the radius of fluorescent haloes. Through the quantification of fluorescence intensity of fluorescein released after the hydrolysis of a fluorescent ester, fluorescein dibutyrate, used as substrate in agar plates, commercial and skimmed milk lipase activity were studied. Moreover, with this method, lipase activity can be monitored in reaction medium that contains compounds which are affected by turbidity or cause measurement interference for UV-spectrophotometer and fluorimeter. In this experiment, boiled skimmed milk was dispersed in the agar gel with fluorescein dibutyrate, and it was used as a reaction medium to mimic natural conditions. The development of such an assay has a potential for applications in industries ranging from pharmaceuticals to food production and monitoring.

The measurement of the urinary excretion of uranium by time-resolved laser-induced fluorescence (TRLIF)

International Nuclear Information System (INIS)

Cavadore, D.; Poirey, B.; Comba, J.B.; Minaud, G.; Ballet, D.

1999-01-01

Rapid direct measurements of the urinary excretion of uranium are often disturbed by metabolic uncertainties and analytical interferences. One consequence of these phenomena is detection limits or uncertainties that are too high. The technique proposed here associates rapid processing of the sample with an optimised measurement system. The objectives of the study - rapidity of response, accuracy and precision lower than 10% and ease of operation - are attained by using a solid power laser as excitation source in conjunction with a modified commercial fluorimeter. We describe the analytical stages for the two methods used (direct measurement and measurement after mineralisation of the sample). The experimental results achieved with 120 measurements are compared with the results obtained by extraction chromatography. The advantages and drawbacks of the TRLIF technique are discussed. Finally, the values of the natural urinary excretion of uranium among 80 non-exposed workers from the Marcoule region are presented as a function of the analytical technique selected. (authors)

Early warning smartphone diagnostics for water security and analysis using real-time pH mapping

Science.gov (United States)

Hossain, Md. Arafat; Canning, John; Ast, Sandra; Rutledge, Peter J.; Jamalipour, Abbas

2015-12-01

Early detection of environmental disruption, unintentional or otherwise, is increasingly desired to ensure hazard minimization in many settings. Here, using a field-portable, smartphone fluorimeter to assess water quality based on the pH response of a designer probe, a map of pH of public tap water sites has been obtained. A custom designed Android application digitally processed and mapped the results utilizing the global positioning system (GPS) service of the smartphone. The map generated indicates no disruption in pH for all sites measured, and all the data are assessed to fall inside the upper limit of local government regulations, consistent with authority reported measurements. This implementation demonstrates a new security concept: network environmental forensics utilizing the potential of novel smartgrid analysis with wireless sensors for the detection of potential disruption to water quality at any point in the city. This concept is applicable across all smartgrid strategies within the next generation of the Internet of Things and can be extended on national and global scales to address a range of target analytes, both chemical and biological.

Oxygen sensor nanoparticles for monitoring bacterial growth and characterization of dose–response functions in microfluidic screenings

International Nuclear Information System (INIS)

Cao, Jialan; Köhler, J. Michael; Nagl, Stefan; Kothe, Erika

2015-01-01

We are presenting a microfluidic droplet-based system for non-invasive, simultaneous optical monitoring of oxygen during bacterial cultivation in nL-sized droplets using ∼350 nm nanobeads made from polystyrene and doped with the NIR-emitting oxygen probe platinum (II) 5, 10, 15, 20-meso-tetraphenyltetrabenzoporphyrin (PtTPTBP). Data were readout by a two-channel micro flow-through fluorimeter and a two-channel micro flow-through photometer. The time-resolved miniaturized optical multi endpoint detection was applied to simultaneously sense dissolved oxygen, cellular autofluorescence, and cell density in nL-sized segments. Two bacterial strains were studied that are resistant to heavy metal ions, viz. Streptomyces acidiscabies E13 and Psychrobacillus psychrodurans UrPLO1. The study has two main features in that it demonstrates (a) the possibility to monitor the changes in oxygen partial pressure during metabolic activity of different bacterial cultures inside droplets, and (b) the efficiency of droplet-based microfluidic techniques along with multi-parameter optical sensing for highly resolved microtoxicological screenings in aquatic systems. (author)

Helicase-dependent amplification of nucleic acids.

Science.gov (United States)

Cao, Yun; Kim, Hyun-Jin; Li, Ying; Kong, Huimin; Lemieux, Bertrand

2013-10-11

Helicase-dependent amplification (HDA) is a novel method for the isothermal in vitro amplification of nucleic acids. The HDA reaction selectively amplifies a target sequence by extension of two oligonucleotide primers. Unlike the polymerase chain reaction (PCR), HDA uses a helicase enzyme to separate the deoxyribonucleic acid (DNA) strands, rather than heat denaturation. This allows DNA amplification without the need for thermal cycling. The helicase used in HDA is a helicase super family II protein obtained from a thermophilic organism, Thermoanaerobacter tengcongensis (TteUvrD). This thermostable helicase is capable of unwinding blunt-end nucleic acid substrates at elevated temperatures (60° to 65°C). The HDA reaction can also be coupled with reverse transcription for ribonucleic acid (RNA) amplification. The products of this reaction can be detected during the reaction using fluorescent probes when incubations are conducted in a fluorimeter. Alternatively, products can be detected after amplification using a disposable amplicon containment device that contains an embedded lateral flow strip. Copyright © 2013 John Wiley & Sons, Inc.

Detection of knockdown resistance (kdr mutations in Anopheles gambiae: a comparison of two new high-throughput assays with existing methods

Directory of Open Access Journals (Sweden)

Ball Amanda

2007-08-01

Full Text Available Abstract Background Knockdown resistance (kdr is a well-characterized mechanism of resistance to pyrethroid insecticides in many insect species and is caused by point mutations of the pyrethroid target site the para-type sodium channel. The presence of kdr mutations in Anopheles gambiae, the most important malaria vector in Africa, has been monitored using a variety of molecular techniques. However, there are few reports comparing the performance of these different assays. In this study, two new high-throughput assays were developed and compared with four established techniques. Methods Fluorescence-based assays based on 1 TaqMan probes and 2 high resolution melt (HRM analysis were developed to detect kdr alleles in An. gambiae. Four previously reported techniques for kdr detection, Allele Specific Polymerase Chain Reaction (AS-PCR, Heated Oligonucleotide Ligation Assay (HOLA, Sequence Specific Oligonucleotide Probe – Enzyme-Linked ImmunoSorbent Assay (SSOP-ELISA and PCR-Dot Blot were also optimized. The sensitivity and specificity of all six assays was then compared in a blind genotyping trial of 96 single insect samples that included a variety of kdr genotypes and African Anopheline species. The relative merits of each assay was assessed based on the performance in the genotyping trial, the length/difficulty of each protocol, cost (both capital outlay and consumable cost, and safety (requirement for hazardous chemicals. Results The real-time TaqMan assay was both the most sensitive (with the lowest number of failed reactions and the most specific (with the lowest number of incorrect scores. Adapting the TaqMan assay to use a PCR machine and endpoint measurement with a fluorimeter showed a slight reduction in sensitivity and specificity. HRM initially gave promising results but was more sensitive to both DNA quality and quantity and consequently showed a higher rate of failure and incorrect scores. The sensitivity and specificity of AS

Detection of knockdown resistance (kdr) mutations in Anopheles gambiae: a comparison of two new high-throughput assays with existing methods

Science.gov (United States)

Bass, Chris; Nikou, Dimitra; Donnelly, Martin J; Williamson, Martin S; Ranson, Hilary; Ball, Amanda; Vontas, John; Field, Linda M

2007-01-01

Background Knockdown resistance (kdr) is a well-characterized mechanism of resistance to pyrethroid insecticides in many insect species and is caused by point mutations of the pyrethroid target site the para-type sodium channel. The presence of kdr mutations in Anopheles gambiae, the most important malaria vector in Africa, has been monitored using a variety of molecular techniques. However, there are few reports comparing the performance of these different assays. In this study, two new high-throughput assays were developed and compared with four established techniques. Methods Fluorescence-based assays based on 1) TaqMan probes and 2) high resolution melt (HRM) analysis were developed to detect kdr alleles in An. gambiae. Four previously reported techniques for kdr detection, Allele Specific Polymerase Chain Reaction (AS-PCR), Heated Oligonucleotide Ligation Assay (HOLA), Sequence Specific Oligonucleotide Probe – Enzyme-Linked ImmunoSorbent Assay (SSOP-ELISA) and PCR-Dot Blot were also optimized. The sensitivity and specificity of all six assays was then compared in a blind genotyping trial of 96 single insect samples that included a variety of kdr genotypes and African Anopheline species. The relative merits of each assay was assessed based on the performance in the genotyping trial, the length/difficulty of each protocol, cost (both capital outlay and consumable cost), and safety (requirement for hazardous chemicals). Results The real-time TaqMan assay was both the most sensitive (with the lowest number of failed reactions) and the most specific (with the lowest number of incorrect scores). Adapting the TaqMan assay to use a PCR machine and endpoint measurement with a fluorimeter showed a slight reduction in sensitivity and specificity. HRM initially gave promising results but was more sensitive to both DNA quality and quantity and consequently showed a higher rate of failure and incorrect scores. The sensitivity and specificity of AS-PCR, SSOP-ELISA, PCR Dot

Evaluation of zinc accumulation, allocation, and tolerance in Zea mays L. seedlings: implication for zinc phytoextraction.

Science.gov (United States)

Bashmakov, Dmitry I; Lukatkin, Alexander S; Anjum, Naser A; Ahmad, Iqbal; Pereira, Eduarda

2015-10-01

This work investigated the accumulation, allocation, and impact of zinc (Zn; 1.0 μM-10 mM) in maize (Zea mays L.) seedlings under simulated laboratory conditions. Z. mays exhibited no significant change in its habitus (the physical characteristics of plants) up to 10-1000 μM of Zn (vs 5-10 mM Zn). Zn tolerance evaluation, based on the root test, indicated a high tolerance of Z. mays to both low and intermediate (or relatively high) concentrations of Zn, whereas this plant failed to tolerate 10 mM Zn and exhibited a 5-fold decrease in its Zn tolerance. Contingent to Zn treatment levels, Zn hampered the growth of axial organs and brought decreases in the leaf area, water regime, and biomass accumulation. Nevertheless, at elevated levels of Zn (10 mM), Zn(2+) was stored in the root cytoplasm and inhibited both axial organ growth and water regime. However, accumulation and allocation of Zn in Z. mays roots, studied herein employing X-ray fluorimeter and histochemical methods, were close to Zn accumulator plants. Overall, the study outcomes revealed Zn tolerance of Z. mays, and also implicate its potential role in Zn phytoextraction.

Photoluminescence behavior of riboflavin and lumiflavin in liquid solutions and solid films

Energy Technology Data Exchange (ETDEWEB)

Penzkofer, A., E-mail: alfons.penzkofer@physik.uni-regensburg.de [Fakultaet fuer Physik, Universitaet Regensburg, Universitaetsstrasse 31, D-93053 Regensburg (Germany)

2012-05-25

Highlights: Black-Right-Pointing-Pointer Delayed fluorescence and phosphorescence spectra of flavins in starch films measured. Black-Right-Pointing-Pointer Quantum yield of singlet-triplet intersystem determined with a new approach. Black-Right-Pointing-Pointer Theory developed for determination of luminescence quantum yields of films. Black-Right-Pointing-Pointer Delayed fluorescence and phosphorescence lifetimes of flavins in starch films measured. Black-Right-Pointing-Pointer Singlet and triplet relevant parameters of riboflavin and lumiflavin determined. - Abstract: The absorption and emission behavior of riboflavin and lumiflavin in water, tetrahydrofuran (THF), water-starch, THF-polystyrene, starch films, and polystyrene films was studied at room temperature. Absorption cross-section spectra, fluorescence quantum distributions, and fluorescence quantum yields were determined. For the starch films additionally phosphorescence and delayed fluorescence spectra as well as phosphorescence lifetimes and delayed fluorescence lifetimes were measured and their quantum yields of intersystem-crossing, intrinsic triplet-based phosphorescence quantum yields, T{sub 1}-S{sub 0} radiative lifetimes, and S{sub 0}-T{sub 1} absorption strengths were calculated. A method of absolute intrinsic luminescence quantum distribution and quantum yield determination for dye doped films on transparent plates with a fluorimeter is described.

Process, product, and waste-stream monitoring with fiber optics

International Nuclear Information System (INIS)

Milanovich, F.P.; Hirschfeld, T.

1983-07-01

Fiber optic technology, motivated by communications and defense applications, has advanced significantly the past ten years. In particular, advances have been made in visible radiation transmission efficiency with concurrent reductions in fiber size, weight, and cost. Researchers at the Lawrence Livermore National Laboratory (LLNL) coupled these advances in fiber optic technology with analytical fluorescence analysis to establish a new technology - remote fiber fluorimetry (RFF). Laser-based RFF offers the potential to measure and monitor from one central and remote laboratory, on-line, and in near real time, trace (ppM) to substantial (g/L) concentrations of selected chemical species in typical process, product, and waste streams. The fluorimeter consists of a fluorescence or Raman spectrometer; unique coupling optics that separates input excitation (laser) radiation from return (fluorescence) radiation; a fiber optic cable; and an optrode - a terminal that interfaces the fiber to the measurement point, which is designed to respond quantitatively to a particular chemical species. At LLNL, research is underway into optrodes that measure pressure, temperature, and pH and those that detect and quantify various actinides, sulfates, inorganic chloride, hydrogen sulfide, aldehydes, and alcohols

DPASV analytical technique for ppb level uranium analysis

Science.gov (United States)

Pal, Sangita; Singha, Mousumi; Meena, Sher Singh

2018-04-01

Determining uranium in ppb level is considered to be most crucial for reuse of water originated in nuclear industries at the time of decontamination of plant effluents generated during uranium (fuel) production, fuel rod fabrication, application in nuclear reactors and comparatively small amount of effluents obtained during laboratory research and developmental work. Higher level of uranium in percentage level can be analyzed through gravimetry, titration etc, whereas inductively coupled plasma-atomic energy spectroscopy (ICP-AES), fluorimeter are well suited for ppm level. For ppb level of uranium, inductively coupled plasma - mass spectroscopy (ICP-MS) or Differential Pulse Anodic Stripping Voltammetry (DPASV) serve the purpose. High precision, accuracy and sensitivity are the crucial for uranium analysis in trace (ppb) level, which are satisfied by ICP-MS and stripping voltammeter. Voltammeter has been found to be less expensive, requires low maintenance and is convenient for measuring uranium in presence of large number of other ions in the waste effluent. In this paper, necessity of uranium concentration quantification for recovery as well as safe disposal of plant effluent, working mechanism of voltammeter w.r.t. uranium analysis in ppb level with its standard deviation and a data comparison with ICP-MS has been represented.

New visible and selective DNA staining method in gels with tetrazolium salts.

Science.gov (United States)

Paredes, Aaron J; Naranjo-Palma, Tatiana; Alfaro-Valdés, Hilda M; Barriga, Andrés; Babul, Jorge; Wilson, Christian A M

2017-01-15

DNA staining in gels has historically been carried out using silver staining and fluorescent dyes like ethidium bromide and SYBR Green I (SGI). Using fluorescent dyes allows recovery of the analyte, but requires instruments such as a transilluminator or fluorimeter to visualize the DNA. Here we described a new and simple method that allows DNA visualization to the naked eye by generating a colored precipitate. It works by soaking the acrylamide or agarose DNA gel in SGI and nitro blue tetrazolium (NBT) solution that, when exposed to sunlight, produces a purple insoluble formazan precipitate that remains in the gel after exposure to light. A calibration curve made with a DNA standard established a detection limit of approximately 180 pg/band at 500 bp. Selectivity of this assay was determined using different biomolecules, demonstrating a high selectivity for DNA. Integrity and functionality of the DNA recovered from gels was determined by enzymatic cutting with a restriction enzyme and by transforming competent cells after the different staining methods, respectively. Our method showed the best performance among the dyes employed. Based on its specificity, low cost and its adequacy for field work, this new methodology has enormous potential benefits to research and industry. Copyright © 2016 Elsevier Inc. All rights reserved.

Analytical Application of Flow Immunosensor in Detection of Thyroxine and Triiodothyronine in Serum.

Science.gov (United States)

Wani, Tanveer A; Zargar, Seema; Majid, Salma; Darwish, Ibrahim A

2016-11-01

In this study, an immunosensor based on kinetic exclusion analysis (KinExA) was used for thyroxine (T4) and triiodothyronine (T3) estimation. A KinExA™ 3200 instrument was used for this analysis, which is an automated flow fluorimeter designed to separate free unbound antibody binding sites in reaction mixtures of antibody, antigen, and antibody-antigen complex. A T3-BSA- and T4-BSA-coated polymethyl methacrylate (PMMA) bead microcolumn is generated inside the flow cell of the instrument. A sample mixture containing T3 and T4 with their respective monoclonal antibodies and their complexes are drawn past the microbead column. The unbound T3 or T4 monoclonal antibody binding sites are captured by their respective T3 and T4 antigens coated on the PMMA beads as bovine serum albumin conjugates. Fluorescently labeled secondary antibodies bind to the T3 or T4 antigen-antibody complex to generate fluorescence intensity for analysis. The limit of detection for the T3 and T4 assays was found to be 0.06 and 1.9 ng mL -1 with acceptable precision values. The convenience of the automated KinExA format may be valuable in medical diagnostic laboratories.

Smart phone based bacterial detection using bio functionalized fluorescent nanoparticles

International Nuclear Information System (INIS)

Rajendran, Vinoth Kumar; Bakthavathsalam, Padmavathy; Ali, Baquir Mohammed Jaffar

2014-01-01

We are describing immunochromatographic test strips with smart phone-based fluorescence readout. They are intended for use in the detection of the foodborne bacterial pathogens Salmonella spp. and Escherichia coli O157. Silica nanoparticles (SiNPs) were doped with FITC and Ru(bpy), conjugated to the respective antibodies, and then used in a conventional lateral flow immunoassay (LFIA). Fluorescence was recorded by inserting the nitrocellulose strip into a smart phone-based fluorimeter consisting of a light weight (40 g) optical module containing an LED light source, a fluorescence filter set and a lens attached to the integrated camera of the cell phone in order to acquire high-resolution fluorescence images. The images were analysed by exploiting the quick image processing application of the cell phone and enable the detection of pathogens within few minutes. This LFIA is capable of detecting pathogens in concentrations as low as 10 5 cfu mL −1 directly from test samples without pre-enrichment. The detection is one order of magnitude better compared to gold nanoparticle-based LFIAs under similar condition. The successful combination of fluorescent nanoparticle-based pathogen detection by LFIAs with a smart phone-based detection platform has resulted in a portable device with improved diagnosis features and having potential application in diagnostics and environmental monitoring. (author)

Feasibility of using fiber optics for monitoring ground water contaminants

International Nuclear Information System (INIS)

Hirschfeld, T.; Deaton, T.; Milanovich, F.; Klainer, S.M.

1984-06-01

The report contains the results of the initial feasibility study for a research program undertaken to develop the technology needed to use fiber optics for monitoring groundwater contaminants. The technology appears especially well suited to the requirements of detection monitoring where a few indicator parameters can be measured continuously by sensors placed down small-diameter monitoring wells. Data are generated at a remote, centrally located fluorimeter, connected to the sampling sites by inexpensive optical fibers. The analytical method is laser-induced fluorescence which gives the desired sensitivity. The optrode, a chemical system and/or a mechanical device at the distal end of a fiber optic, furnishes the needed specificity. Various fiber and optrode configurations have been evaluated and their applications to groundwater monitoring are discussed. Feasibility is shown for physical measurements such as temperature, pressure and pH. Chemical detection and quantification of the actinides, inorganic and organic chlorides, sulfates, alcohols, aldehydes, pesticides and tracer materials are presented. Finally, it is shown that the need for smaller diameter wells (as compared to conventional sampling methods), and the ability to make up to 50 unattended in situ measurements, using a reasonably priced centralized fluorometer system connected to the sampling sites by inexpensive optical fibers, results in acceptable economy

Role of maintenance of analytical instruments in the proceedings of quality control laboratory

International Nuclear Information System (INIS)

Haribabu, A.; Sailoo, C.C.; Balaji Rao, Y.; Subba Rao, Y.

2015-01-01

Control Laboratory being a centralized analytical facility of Nuclear Fuel Complex (NFC) is engaged in chemical qualification of all nuclear materials processed/produced at NFC. The primary responsibility of control laboratory is to provide timely analytical results of raw materials, intermediates and final products to all the production plants of NFC for downstream processing. Annual analytical load of nearly five lakhs of estimations are being carried out at laboratory. For this purpose a gamut of analytical facilities ranging from classical methods like gravimetry, volumetry etc. to fully automated state-of-art analytical instruments like ICP-AES, Gas Analysers, Flame and Graphite Furnace-AAS, Direct Reading Emission Spectrometer (DRES), RF GD-OES, TIMS, WD-XRFS, ED-XRFS, Laser based PSD Analyser, Laser Fluorimeter, UV-Vis Spectrophotometer, Gamma Ray Spectrometer, Ion-Chromatography, Gas Chromatography are used to acquire analytical data to see the suitability of products for their intended use. Depending on the applications, analysts validate their procedures, calibrate their instruments, and perform additional instrument checks, such as system suitability tests and analysis of in-process quality control check samples. With the increasing sophistication and automation of analytical instruments, an increasing demand has been placed on maintenance engineers to qualify these instruments for the purpose

Studies on effective decomposition of monazite minerals by variety of phosphate fluxes for simple and direct determination of uranium by LED fluorimeter

International Nuclear Information System (INIS)

Gopal, Leela; Hanuman, V.V.; Chakrapani, G.

2013-01-01

A simple, rapid, effective sample decomposition method is developed for the determination of uranium (U) in monazite minerals by fluorimetric (Light Emitting Diodes (LED) based) technique. The salts of sodium dihydrogen phosphate (NaH 2 PO 4 ), disodium hydrogen phosphate (Na 2 HPO 4 ) and tetrasodium pyrophosphate (Na 4 P 2 O 7 ) were used to conduct studies on effective decomposition and dissolution of monazite minerals. The flux short listed for sample decomposition has several advantages. The fusion is very simple (involve minimal skills), time saving and eco-friendly (no acids were used for sample dissolution), where as in the reported conventional sample decomposition methods involving fusion with sodium peroxide, mixture of KHF 2 and NaF, mineral acids are being used for sample decomposition to get clear solution. Further this solution cannot be used directly for uranium determination by LED fluorimetry; hence separation is required, resulting in low sample throughput. In the present method no such separation is required as the flux itself acts as a fluorescence enhancing reagent and buffer (maintaining the optimum pH of 7.1 ± 0.1). The fused melt of the flux mixture when dissolved in water, gives clear and stable solution. The accuracy and precision of the method was evaluated by analyzing Certified Reference Material, IGS-36 (Institute of Geological Sciences, UK) and monazite samples received from BSOI, Trivandrum. The accuracy of the data was further evaluated by comparing with conventional standard decomposition methods. The results are well within experimental error. RSD of the method is ±2% at 0.30% U 3 O 8 in monazite minerals. (author)

A polymeric dosimeter film based on optically-stimulated luminescence for dose measurements below 1 kGy

International Nuclear Information System (INIS)

Kovacs, A.; Baranyai, M.; Wojnarovits, L.; Slezsak, I.; McLaughlin, W.L.; Miller, S.D.; Miller, A.; Fuochi, P.G.; Lavalle, M.

1999-01-01

A new potential dosimetry system 'Sunna' containing a microcrystalline dispersion of an optically-stimulated fluor in a plastic matrix has been recently developed to measure and image high doses. Our previous investigations have revealed that the new dosimeter system is capable of measuring absorbed doses in the dose range of 1-100 kGy. The optically-stimulated luminescence (OSL) analysis is based on the blue light stimulation of the colour center states produced upon irradiation, and the intensity of the resulting red-light emission is used to measure absorbed dose. This analysis is carried out with a simple table-top fluorimeter developed for this purpose having also the ability to calculate the mathematical formula of the calibration function. The Sunna dosimeter was recently investigated for potential use in lower dose range below 1 kGy. These investigations have shown that the film is suitable for measuring doses in the range of 1-1000 Gy for both electron and gamma radiation. To test the applicability of the film, its reproducibility, stability, sensitivity to ambient and UV light and irradiation temperature were measured. The stability of the dosimeter was investigated by monitoring the change of the OSL signal with storage time after irradiation. Further experiments proved the homogeneity of the film with respect to thickness variation, and limited differences in its response were found between batches. (author)

Stimulation and inhibition of bacterial growth by caffeine dependent on chloramphenicol and a phenolic uncoupler--a ternary toxicity study using microfluid segment technique.

Science.gov (United States)

Cao, Jialan; Kürsten, Dana; Schneider, Steffen; Köhler, J Michael

2012-10-01

A droplet-based microfluidic technique for the fast generation of three dimensional concentration spaces within nanoliter segments was introduced. The technique was applied for the evaluation of the effect of two selected antibiotic substances on the toxicity and activation of bacterial growth by caffeine. Therefore a three-dimensional concentration space was completely addressed by generating large sequences with about 1150 well separated microdroplets containing 216 different combinations of concentrations. To evaluate the toxicity of the ternary mixtures a time-resolved miniaturized optical double endpoint detection unit using a microflow-through fluorimeter and a two channel microflow-through photometer was used for the simultaneous analysis of changes on the endogenous cellular fluorescence signal and on the cell density of E. coli cultivated inside 500 nL microfluid segments. Both endpoints supplied similar results for the dose related cellular response. Strong non-linear combination effects, concentration dependent stimulation and the formation of activity summits on bolographic maps were determined. The results reflect a complex response of growing bacterial cultures in dependence on the combined effectors. A strong caffeine induced enhancement of bacterial growth was found at sublethal chloramphenicol and sublethal 2,4-dinitrophenol concentrations. The reliability of the method was proved by a high redundancy of fluidic experiments. The results indicate the importance of multi-parameter investigations for toxicological studies and prove the potential of the microsegmented flow technique for such requirements.

Experimental studies on the weathering of chemicals in a field trial to predict their behaviour in case of a spill

International Nuclear Information System (INIS)

Mamaca, E.; Merlin, F.X.; Le Floch, S.

2004-01-01

Most of the world's production of vegetable oil is transported by sea. In 2001, nearly 850,000 tons of vegetable oil entered and left harbours in France. This trend increases the risk of accidental spills at sea. The physical state of vegetable oil changes when it is spilled at sea, turning this non-toxic product into a pollutant that damages the marine ecosystem. This study demonstrated how vegetable oil could react when spilled at sea. A series of field studies were conducted to obtain experimental data on the behaviour of vegetable oil both on the surface of water and in the water column. Castor oil, soybean oil, oleic acid and dioctylphtalate were released at sea and the dispersion of the oil in the water was monitored with a fluorimeter. Measurements were taken to a depth of 1 metre. Emulsification and viscosity kinetics were monitored. The study showed that the behaviour of the 4 products depends on the nature of the product and weather conditions such as wind and sea surface state. Vegetable oil spilled at sea behaves differently from spilled chemical products in terms of solubility. It was suggested that in the case of an accidental spill at sea, emergency responders should first pump the oil and then use dispersants. 6 refs., 2 tabs., 11 figs

Ultratrace analysis of actinides via coprecipitation/laser-induced fluorescence spectroscopy

International Nuclear Information System (INIS)

Miller, S.M.

1982-01-01

Actinides were selectively preconcentrated by coprecipitating each out of solution with a fluoride matrix and calcining each sample at 800 0 C. The fluorescence spectrum of each sample was recorded by illuminating the sample with laser light and detecting fluorescence with either a fluorescence/Raman spectrometer, an infrared spectrometer or in certain cases a filter fluorimeter. Three previously unobserved actinide spectra were recorded. Narrow lines at 546.9 nm, 564.6 nm, and 569.6 nm were found for CaF 2 :PuO 2++ at 10K. CaF 2 :Am + 3 displayed two broadband fluorescent peaks at 625 nm and 746 nm at room temperature and CaF 2 :Pu + 3 possessed a fluorescent peak at 1.22 microns at 10K. Energy transfer was observed in the form of Tb fluorescence quenching in TbF 3 :Pu + 3 when Pu was present in quantities of 10 ppM or more and in the form of Tb fluorescence enhancement in TbF 3 :Am + 3 when 1 ppM or more of Am was present. Careful sample preparation and the use of temporal as well as a spectral discrimination system extended the detection limit of U from 1 ml samples to the subfemtogram level. The fluorescence detection limits for Pu and Am were extended to 0.48 and 0.032 pg/ml. 39 figures, 9 tables

On-site monitoring of Hebei Spirit oil spill by fluorometric detection of oil residues in coastal waters off Taean, Korea

Energy Technology Data Exchange (ETDEWEB)

Kim, M.; Yim, U.H.; Hong, S.H.; Jung, J.H.; Won, J.; An, J.; Choi, H.W.; Shim, W.J. [Korea Ocean Research and Development Inst., Geoje (Korea, Republic of)

2009-07-01

This paper discussed activities conducted to monitor a crude oil spill that contaminated over 70 km of the western Korean coastline. Contamination levels and temporal variations of dissolved and dispersed oils in sea and pore water at 40 beaches were monitored using a portable fluorimeter for 10 months after the spill. More than 980 samples from the heavily-impacted Mallipo Beach area were analyzed. The analysis showed that oil concentrations in the sea water were as high as 16,600 {mu}g/L directly after the spill, and decreased to below the Korean marine water quality standard of 10 {mu}g/L at most sites 10 months after the spill. However, the oil content in pore water remained high, with levels of up to 2,320 {mu}g/L for the first few months following the spill. Higher oil contamination levels were observed at some sites for up to 10 months after the spill. Results of the study suggested that oil in pore water persisted in confined areas along the coastline. Results from the fluorescence detection technique were then compared with traditional gas chromatography (GC) techniques of total petroleum hydrocarbon analysis. It was concluded that fluorescence detection was capable of generating accurate results more quickly and cost-effectively than traditional GC techniques. 22 refs., 7 figs.

Spatial distribution of uranium and basic water quality parameter in the capital of Bihar and consequent ingestion dose.

Science.gov (United States)

Kumar, Deepak; Singh, Anshuman; Jha, Rishi Kumar

2018-04-21

Investigation of presence of Uranium (U) in groundwater/drinking water is an active are of research due to its chemical and radiological toxicity as well as long-term health effects. The current study had the objective of estimating U as a naturally occurring radioactive element in groundwater samples and assessment of ingestion dose, when groundwater is the source of drinking water. The random sampling method was chosen for the collection of samples based on population density. The estimation of U was done using LED fluorimeter. Statistical tools were applied to analyze the data and its spatial distribution. The U concentrations in three blocks of urban Patna were well below the permissible limits suggested by different health agencies of the world. A correlation test was performed to analyze the association of U with other physiochemical parameters of water samples. It was found that the sulfate, chloride, calcium, hardness, alkalinity, TDS, salinity, and ORP were positively correlated, whereas fluoride, phosphate, magnesium, dissolved oxygen, and pH were negatively correlated with U concentrations. The ingestion dose due to U, occurring in groundwater, was found to vary from 0.2-27.0 μSv y -1 with a mean of 4.2 μSv y - 1 , which was well below the recommended limit of 0.1 mSv (WHO WHO Chron 38:104-108, 2012).Therefore, the water in this region is fit for drinking purposes.

Surface defect assisted broad spectra emission from CdSe quantum dots for white LED application

Science.gov (United States)

Samuel, Boni; Mathew, S.; Anand, V. R.; Correya, Adrine Antony; Nampoori, V. P. N.; Mujeeb, A.

2018-02-01

This paper reports, broadband photoluminescence from CdSe quantum dots (QDs) under the excitation of 403 nm using fluorimeter and 403 nm CW laser excitation. The broad spectrum obtained from the colloidal quantum dots was ranges from 450 nm to 800 nm. The broadness of the spectra was attributed to the merging of band edge and defect driven emissions from the QDs. Six different sizes of particles were prepared via kinetic growth method by using CdO and elemental Se as sources of Cd and Se respectively. The particle sizes were measured from TEM images. The size dependent effect on broad emission was also studied and the defect state emission was found to be predominant in very small QDs. The defect driven emission was also observed to be redshifted, similar to the band edge emission, due to quantum confinement effect. The emission corresponding to different laser power was also studied and a linear relation was obtained. In order to study the colour characteristics of the emission, CIE chromaticity coordinate, CRI and CCT of the prepared samples were measured. It is observed that, these values were tunable by the addition of suitable intensity of blue light from the excitation source to yield white light of various colour temperatures. The broad photoluminescence spectrum of the QDs, were compared with that of a commercially available white LED. It was found that the prepared QDs are good alternatives for the phosphor in phosphor converted white LEDs, to provide good spectral tunability.

Fluorimetric method for determination of Beryllium

International Nuclear Information System (INIS)

Sparacino, N.; Sabbioneda, S.

1996-10-01

The old fluorimetric method for the determination of Beryllium, based essentially on the fluorescence of the Beryllium-Morine complex in a strongly alkaline solution, is still competitive and stands the comparison with more modern methods or at least three reasons: in the presence of solid or gaseous samples (powders), the times necessary to finalize an analytic determination are comparable since the stage of the process which lasts the longest is the mineralization of the solid particles containing Beryllium, the cost of a good fluorimeter is by far Inferior to the cost, e. g., of an Emission Spectrophotometer provided with ICP torch and magnets for exploiting the Zeeman effect and of an Atomic absorption Spectrophotometer provided with Graphite furnace; it is possible to determine, fluorimetrically, rather small Beryllium levels (about 30 ng of Beryllium/sample), this potentiality is more than sufficient to guarantee the respect of all the work safety and hygiene rules now in force. The study which is the subject of this publication is designed to the analysis procedure which allows one to reach good results in the determination of Beryllium, chiefly through the control and measurement of the interference effect due to the presence of some metals which might accompany the environmental samples of workshops and laboratories where Beryllium is handled, either at the pure state or in its alloys. The results obtained satisfactorily point out the merits and limits of this analytic procedure

Retail Sugar From One Zambian Community Does Not Meet Statutory Requirements for Vitamin A Fortification.

Science.gov (United States)

Greene, Matthew D; Kabaghe, Gladys; Musonda, Mofu; Palmer, Amanda C

2017-12-01

Industrial food fortification is a major strategy to improve dietary micronutrient intakes and prevent deficiencies. Zambia introduced mandatory sugar fortification with vitamin A, at a target of 10 mg/kg, in 1998. Representative surveys conducted since that time do not support marked improvement in vitamin A status. To describe vitamin A concentrations in retail sugar, as well as vendor practices, perceptions of fortified foods, and sugar use practices. We conducted a census of sugar vendors in one Zambian community, capturing information on vendors, available brands and packaging options, and storage conditions. We purchased all brands and package types of sugar available at each vendor. In a 15% subsample, we conducted semi-structured interviews with vendor-consumer pairs. We tested 50% of sugar samples at random for vitamin A using an iCheck portable fluorimeter. The distribution of vitamin A in sugar in market samples was highly skewed, with a median of 3.1 mg/kg (25th-75th percentiles: 1.8-5.5) and a range from 0.2 to 29.9 mg/kg. Only 11.3% of samples met the 10 mg/kg statutory requirement. Sugar was primarily repackaged and sold in small quantities, with rapid turnover of stocks. Perceptions of fortification by vendors and consumers were generally positive. Vitamin A in fortified sugar fell well below statutory requirements. Our data point to challenges at regional depot and/or poor adherence to fortification standards at the factory level. A renewed commitment to monitoring and enforcement will be required for Zambia to benefit from a food fortification strategy.

Uranium exploration of Samar Island

International Nuclear Information System (INIS)

Santos, G. Jr.

1979-02-01

Uranium exploration is being undertaken to meet the requirements of the Philippine Nuclear Power Plant-1 (PNPP-1) programmed to operate in 1982, for about 140 metric tons annually or 2664 MT of U 3 O 8 up to the year 2000. Samar was chosen as the survey pilot project and the method used was a geochemical reconnaissance or low density observation survey to delineate broad areas where follow-up uranium surveys may be undertaken. Stream sediments or surface waters were collected at each sampling point at a density of one sample per 20-25 sq. km. The conductance and pH of the water were measured with a conductivity meter and pH respectively. Radioactivity was determined using a portable scintillometer. The stream sediment and heavy mineral samples were analyzed for uranium (U), copper (CCu), lead (Pb), zinc (Zn), manganese (Mn), silver (Ag), cobalt (Co), nickel (Ni). Water samples were analyzed for uranium only. The solid samples were digested in an acid mixture of 85% concentrated nitric acid and 15% concentrated hydrochloric acid, and the leachable uranium was determined using a fluorimeter. The detection limits for uranium were 0.3 ppb and 0.3 ppm for water and solid samples, respectively. Analysis for Cu, Pb, Zn, Mn, Ag, Co, and Ni were done by Atomic Absorption Spectrophotometry using the same leaching solution prepared for uranium analysis. Over 9000 determinations were done on nearly 1600 samples. The survey delineated at least two areas where follow-up surveys for uranium are warranted. These areas are the San Isidro - Catarman in Northwestern Samar, and the vicinity of Bagacay mines in Central Samar

Early development and characterization of a DNA-based radiation dosimeter

Science.gov (United States)

Avarmaa, Kirsten A.

It is the priority of first responders to minimize damage to persons and infrastructure in the case of a nuclear emergency due to an accident or deliberate terrorist attack -- if this emergency includes a radioactive hazard, first responders require a simple-to-use, accurate and complete dosimeter for radiation protection purposes in order to minimize the health risk to these individuals and the general population at large. This work consists of the early evaluation of the design and performance of a biologically relevant dosimeter which uses DNA material that can respond to the radiation of any particle type. The construct consists of fluorescently tagged strands of DNA. The signalling components of this dosimeter are also investigated for their sensitivity to radiation damage and light exposure. The dual-labelled dosimeter that is evaluated in this work gave a measurable response to gamma radiation at dose levels of 10 Gy for the given detector design and experimental setup. Further testing outside of this work confirmed this finding and indicated a working range of 100 mGy to 10 Gy using a custom-built fluorimeter as part of a larger CRTI initiative. Characterization of the chromatic components of the dosimeter showed that photobleaching is not expected to have an effect on dosimeter performance, but that radiation can damage the non-DNA signalling components at higher dose levels, although this damage is minimal at lower doses over the expected operating ranges. This work therefore describes the early steps in the quantification of the behaviour of the DNA dosimeter as a potential biologically-based device to measure radiation dose.

A comparative study of distribution coefficients (Kd) for naturally occurring Uranium (U) and Thorium (Th) in two different aquatic environments

International Nuclear Information System (INIS)

Kumar, Ajay; Karpe, Rupali; Rout, Sabyasachi; Narayanan, Usha; Ravi, P.M.

2012-01-01

The uranium and thorium contents and their mobility in aqueous systems are mainly controlled by the pH, alkalinity, the oxidation reduction potential (ORP) and the type of complexing agents present, such as carbonates, phosphates, vanadates, fluorides, sulfates and silicates, etc. A comparative study of distribution coefficients (K d ) for U and Th in sediment-seawater and soil-groundwater system has been carried out. K d was determined using a batch method. In this method, 5 g dried sediment samples was placed in each of seven empty conical flasks and equilibrated for 7 days with 150 mL of sea water containing 10, 20, 30, 40, 50, 75 and 100 mg/L of uranium and thorium contents followed by shaking using end-over end shaker at 25°C. After equilibration time, the samples of each set were centrifuged, filtered through 0.45 μm filter paper and supernatant analyzed for uranium and thorium. In the similar way, experiments were conducted for soil-groundwater system. The concentration of uranium in aliquots of equilibrium solution was measured using laser fluorimeter and Th was determined using anion exchange column followed by co-precipitation with ferric hydroxide and estimated by gross alpha counter. Physico-chemical parameters of soil, sediments, seawater and groundwater were also studied. In this study, K d values have been reported as the mean from two sets of experimental determinations. Based on the resulting data set, it can be concluded that K d values of uranium and thorium are not only dependent on properties of adsorbed phases but also on the kinds of minerals present in that medium. The results of K d values obtained indicated that the sediments have better sorption properties than soil

A study of Uranium contamination and the expected hazards in southwest Punjab

International Nuclear Information System (INIS)

Kumar, Mukesh; Prasher, Sangeeta; Singh, Surinder

2011-01-01

In recent years, concerns are being raised about the possible carcinogenicity and neurological disorders due to the ingested uranium through drinking water in southwest Punjab. Germany's Microtrace Mineral Lab has confirmed that the high level of Uranium and other dangerous heavy metals present in drinking water samples from the region is responsible for 80% of the neurologically disabled children. The present study deals with the analysis of uranium in the water samples collected from Bathinda District of Punjab. The investigations reveals that the significant percentage of the area surveyed has uranium concentrations in the ground water more than the WHO (World Health Organization) recommended limit of 15 g/L. The highest concentration of uranium 100 g/L has been investigated in the water sample from Giana village and the minimum uranium concentration value of 1.84 μg/l is found in Malkana. The average uranium concentration in water samples is found to vary from 9.330.06 μg/l in Rampura to 56. 950.13 μg/l in Bucho Mandi villages. In order to be sure about the results, a few selected samples have been re-examined using laser fluorimeter technique in the Environmental Survey Laboratory, Rajasthan Atomic Power Station of Bhabha Atomic Research Center at Kota, Rajasthan, India. The results for uranium concentration obtained using both techniques are found in fair agreement with some exceptions. The excess cancer risk due to such a high content of uranium in drinking water is found to vary from 25-154x 10 -4 % (1: 3974 -1: 6511) with a mean value 77x, 10 -4 %, which means that at least one individual out of 13026 may be influenced by cancer. (author)

Gestion de la fertilisation azotée des cultures de plein champ. Perspectives d'amélioration de l'efficience d'utilisation de l'azote sur base du suivi du statut azoté de la biomasse aérienne

Directory of Open Access Journals (Sweden)

Goffart, JP.

2013-01-01

Full Text Available Nitrogen fertilization management of open-field crops. Opportunities for improvement of nitrogen use efficiency based on crop nitrogen status monitoring. The improvement of nitrogen use efficiency in crops is currently an important issue in farming due to current and future economical and environmental constraints. Splitting of N fertilizer application is the most suitable approach for providing an optimal match of N need and supply. The implementation of monitoring methods to assess crop N status is required to define the relevant amounts and periods required for split applications. This paper discusses the available methods of split N fertilizer application and their mode of use and implementation. After a short overview of the concept of crop N status, the main existing methods to estimate nitrogen needs are examined for their accuracy, specificity and sensitivity based on research conducted at CRA-W (Production and Sectors Department, Crop Production Systems Unit over the last two decades, focusing specifically on the potato crop. The methods and the results described relate to the petiole sap nitrate concentration test assessed via a reflectometer, the measurement of leaf chlorophyll concentration with a handheld chlorophyll meter, leaf chlorophyll fluorescence readings with handheld fluorimeters, and the measurement of crop light reflectance with a handheld radiometer (ground-based remote sensing or with satellite imagery (spatial remote sensing. Conditions for implementing such methods within decision support systems are briefly described, by focusing on the need to use relative values rather than raw values taken from the readings, and also on the requirement for threshold value definition. The integration of crop nitrogen status values into N fertilization recommendation models is illustrated through a Decision Support System created at CRA-W for the potato crop.

Environmental drivers of phototrophic biofilms in an Alpine show cave (SW-Italian Alps)

International Nuclear Information System (INIS)

Piano, E.; Bona, F.; Falasco, E.; La Morgia, V.; Badino, G.; Isaia, M.

2015-01-01

The proliferation of lampenflora is a major threat for the conservation of show caves, since phototrophic organisms cause physical, chemical and aesthetic damage to speleothems. In this paper we examine the environmental factors influencing the presence and the growth of the three main photosynthetic groups composing phototrophic biofilms in the Bossea show cave (SW-Italian Alps). The presence and the primary production of cyanobacteria, diatoms and green algae were detected with BenthoTorch®, an instrument for in situ measurement of chlorophyll a concentration that has never been used before in caves. By means of different techniques of regression analysis, we highlighted the response of the three photosynthetic groups to different environmental factors. Illuminance proved to be the main factor influencing positively both the probability of the presence and the productivity of the three groups. The presence of seeping water on the substrate and the distance from the cave entrance proved to play an important role in determining patterns of colonization. By means of GIS techniques, we provide thematic maps of the cave, providing a representation of pattern of the density of the three examined photosynthetic groups within different areas of the cave. The same approach may apply to other show caves, aiming at providing suggestions for the cave management (i.e. cleaning of the cave walls and positioning of artificial lights) and reduce impact caused by tourism. - Highlights: • We used a PAM fluorimeter on autotrophic biofilms in a show cave for the first time. • We modelled the environmental factors influencing phototrophic biofilms. • Illuminance, moisture and distance from the entrance proved to be significant. • We produced thematic maps illustrating our results. • We provide suggestions for cave management

Real-Time Fluorescence Measurements of ROS and [Ca2+] in Ischemic / Reperfused Rat Hearts: Detectable Increases Occur only after Mitochondrial Pore Opening and Are Attenuated by Ischemic Preconditioning.

Science.gov (United States)

Andrienko, Tatyana; Pasdois, Philippe; Rossbach, Andreas; Halestrap, Andrew P

2016-01-01

Mitochondrial permeability transition pore (mPTP) opening is critical for ischemia / reperfusion (I/R) injury and is associated with increased [Ca2+] and reactive oxygen species (ROS). Here we employ surface fluorescence to establish the temporal sequence of these events in beating perfused hearts subject to global I/R. A bespoke fluorimeter was used to synchronously monitor surface fluorescence and reflectance of Langendorff-perfused rat hearts at multiple wavelengths, with simultaneous measurements of hemodynamic function. Potential interference by motion artefacts and internal filtering was assessed and minimised. Re-oxidation of NAD(P)H and flavoproteins on reperfusion (detected using autofluorescence) was rapid (t0.5 ROS increases during early reperfusion. However, two different fluorescent cytosolic ROS probes did detect ROS increases after 2-3 min of reperfusion, which was shown to be after initiation of mPTP opening. Cyclosporin A (CsA) and IP attenuated these responses and reduced infarct size. [Ca2+]i (monitored with Indo-1) increased progressively during ischemia, but dropped rapidly within 90 s of reperfusion when total mitochondrial [Ca2+] was shown to be increased. These early changes in [Ca2+] were not attenuated by IP, but substantial [Ca2+] increases were observed after 2-3 min reperfusion and these were prevented by both IP and CsA. Our data suggest that the major increases in ROS and [Ca2+] detected later in reperfusion are secondary to mPTP opening. If earlier IP-sensitive changes occur that might trigger initial mPTP opening they are below our limit of detection. Rather, we suggest that IP may inhibit initial mPTP opening by alternative mechanisms such as prevention of hexokinase 2 dissociation from mitochondria during ischemia.

Sinking towards destiny: High throughput measurement of phytoplankton sinking rates through time-resolved fluorescence plate spectroscopy.

Science.gov (United States)

Bannon, Catherine C; Campbell, Douglas A

2017-01-01

Diatoms are marine primary producers that sink in part due to the density of their silica frustules. Sinking of these phytoplankters is crucial for both the biological pump that sequesters carbon to the deep ocean and for the life strategy of the organism. Sinking rates have been previously measured through settling columns, or with fluorimeters or video microscopy arranged perpendicularly to the direction of sinking. These side-view techniques require large volumes of culture, specialized equipment and are difficult to scale up to multiple simultaneous measures for screening. We established a method for parallel, large scale analysis of multiple phytoplankton sinking rates through top-view monitoring of chlorophyll a fluorescence in microtitre well plates. We verified the method through experimental analysis of known factors that influence sinking rates, including exponential versus stationary growth phase in species of different cell sizes; Thalassiosira pseudonana CCMP1335, chain-forming Skeletonema marinoi RO5A and Coscinodiscus radiatus CCMP312. We fit decay curves to an algebraic transform of the decrease in fluorescence signal as cells sank away from the fluorometer detector, and then used minimal mechanistic assumptions to extract a sinking rate (m d-1) using an RStudio script, SinkWORX. We thereby detected significant differences in sinking rates as larger diatom cells sank faster than smaller cells, and cultures in stationary phase sank faster than those in exponential phase. Our sinking rate estimates accord well with literature values from previously established methods. This well plate-based method can operate as a high throughput integrative phenotypic screen for factors that influence sinking rates including macromolecular allocations, nutrient availability or uptake rates, chain-length or cell size, degree of silification and progression through growth stages. Alternately the approach can be used to phenomically screen libraries of mutants.

Environmental drivers of phototrophic biofilms in an Alpine show cave (SW-Italian Alps)

Energy Technology Data Exchange (ETDEWEB)

Piano, E., E-mail: elena.piano@unito.it [Department of Life Sciences and Systems Biology, University of Turin, Via Accademia Albertina 13, 10123 Turin (Italy); Bona, F.; Falasco, E. [Department of Life Sciences and Systems Biology, University of Turin, Via Accademia Albertina 13, 10123 Turin (Italy); La Morgia, V. [ISPRA, via Ca' Fornacetta, 9, 40064 Ozzano dell' Emilia (Italy); Badino, G. [Department of Physics, University of Turin, Via P. Giuria 1, 10125 Turin (Italy); Isaia, M. [Department of Life Sciences and Systems Biology, University of Turin, Via Accademia Albertina 13, 10123 Turin (Italy)

2015-12-01

The proliferation of lampenflora is a major threat for the conservation of show caves, since phototrophic organisms cause physical, chemical and aesthetic damage to speleothems. In this paper we examine the environmental factors influencing the presence and the growth of the three main photosynthetic groups composing phototrophic biofilms in the Bossea show cave (SW-Italian Alps). The presence and the primary production of cyanobacteria, diatoms and green algae were detected with BenthoTorch®, an instrument for in situ measurement of chlorophyll a concentration that has never been used before in caves. By means of different techniques of regression analysis, we highlighted the response of the three photosynthetic groups to different environmental factors. Illuminance proved to be the main factor influencing positively both the probability of the presence and the productivity of the three groups. The presence of seeping water on the substrate and the distance from the cave entrance proved to play an important role in determining patterns of colonization. By means of GIS techniques, we provide thematic maps of the cave, providing a representation of pattern of the density of the three examined photosynthetic groups within different areas of the cave. The same approach may apply to other show caves, aiming at providing suggestions for the cave management (i.e. cleaning of the cave walls and positioning of artificial lights) and reduce impact caused by tourism. - Highlights: • We used a PAM fluorimeter on autotrophic biofilms in a show cave for the first time. • We modelled the environmental factors influencing phototrophic biofilms. • Illuminance, moisture and distance from the entrance proved to be significant. • We produced thematic maps illustrating our results. • We provide suggestions for cave management.

Real-Time Fluorescence Measurements of ROS and [Ca2+] in Ischemic / Reperfused Rat Hearts: Detectable Increases Occur only after Mitochondrial Pore Opening and Are Attenuated by Ischemic Preconditioning.

Directory of Open Access Journals (Sweden)

Tatyana Andrienko

Full Text Available Mitochondrial permeability transition pore (mPTP opening is critical for ischemia / reperfusion (I/R injury and is associated with increased [Ca2+] and reactive oxygen species (ROS. Here we employ surface fluorescence to establish the temporal sequence of these events in beating perfused hearts subject to global I/R. A bespoke fluorimeter was used to synchronously monitor surface fluorescence and reflectance of Langendorff-perfused rat hearts at multiple wavelengths, with simultaneous measurements of hemodynamic function. Potential interference by motion artefacts and internal filtering was assessed and minimised. Re-oxidation of NAD(PH and flavoproteins on reperfusion (detected using autofluorescence was rapid (t0.5 < 15 s and significantly slower following ischemic preconditioning (IP. This argues against superoxide production from reduced Complex 1 being a critical mediator of initial mPTP opening during early reperfusion. Furthermore, MitoPY1 (a mitochondria-targeted H2O2-sensitive fluorescent probe and aconitase activity measurements failed to detect matrix ROS increases during early reperfusion. However, two different fluorescent cytosolic ROS probes did detect ROS increases after 2-3 min of reperfusion, which was shown to be after initiation of mPTP opening. Cyclosporin A (CsA and IP attenuated these responses and reduced infarct size. [Ca2+]i (monitored with Indo-1 increased progressively during ischemia, but dropped rapidly within 90 s of reperfusion when total mitochondrial [Ca2+] was shown to be increased. These early changes in [Ca2+] were not attenuated by IP, but substantial [Ca2+] increases were observed after 2-3 min reperfusion and these were prevented by both IP and CsA. Our data suggest that the major increases in ROS and [Ca2+] detected later in reperfusion are secondary to mPTP opening. If earlier IP-sensitive changes occur that might trigger initial mPTP opening they are below our limit of detection. Rather, we suggest that

Development of a biosensor protein bullet as a fluorescent method for fast detection of Escherichia coli in drinking water.

Directory of Open Access Journals (Sweden)

Ignacio Gutiérrez-Del-Río

Full Text Available Drinking water can be exposed to different biological contaminants from the source, through the pipelines, until reaching the final consumer or industry. Some of these are pathogenic bacteria and viruses which may cause important gastrointestinal or systemic diseases. The microbiological quality of drinking water relies mainly in monitoring three indicator bacteria of faecal origin, Escherichia coli, Enterococcus faecalis and Clostridium perfringens, which serve as early sentinels of potential health hazards for the population. Here we describe the analysis of three chimeric fluorescent protein bullets as biosensor candidates for fast detection of E. coli in drinking water. Two of the chimeric proteins (based on GFP-hadrurin and GFP-pb5 chimera proteins failed with respect to specificity and/or sensitivity, but the GFP-colS4 chimera protein was able to carry out specific detection of E. coli in drinking water samples in a procedure encompassing about 8 min for final result and this biosensor protein was able to detect in a linear way between 20 and 103 CFU of this bacterium. Below 20 CFU, the system cannot differentiate presence or absence of the target bacterium. The fluorescence in this biosensor system is provided by the GFP subunit of the chimeric protein, which, in the case of the better performing sensor bullet, GFP-colS4 chimera, is covalently bound to a flexible peptide bridge and to a bacteriocin binding specifically to E. coli cells. Once bound to the target bacteria, the excitation step with 395 nm LED light causes emission of fluorescence from the GFP domain, which is amplified in a photomultiplier tube, and finally this signal is converted into an output voltage which can be associated with a CFU value and these data distributed along mobile phone networks, for example. This method, and the portable fluorimeter which has been developed for it, may contribute to reduce the analysis time for detecting E. coli presence in drinking

Adenoviral gene delivery to primary human cutaneous cells and burn wounds.

Science.gov (United States)

Hirsch, Tobias; von Peter, Sebastian; Dubin, Grzegorz; Mittler, Dominik; Jacobsen, Frank; Lehnhardt, Markus; Eriksson, Elof; Steinau, Hans-Ulrich; Steinstraesser, Lars

2006-01-01

The adenoviral transfer of therapeutic genes into epidermal and dermal cells is an interesting approach to treat skin diseases and to promote wound healing. The aim of this study was to assess the in vitro and in vivo transfection efficacy in skin and burn wounds after adenoviral gene delivery. Primary keratinocytes (HKC), fibroblasts (HFB), and HaCaT cells were transfected using different concentrations of an adenoviral construct (eGFP). Transfection efficiency and cytotoxicity was determined up to 30 days. Expression was quantified by FACS analysis and fluorimeter. Cytotoxicity was measured using the trypan blue exclusion method. 45 male Sprague Dawley rats received 2x10(8) pfu of Ad5-CMV-LacZ or carrier control intradermally into either superficial partial thickness scald burn or unburned skin. Animals were euthanized after 48 h, 7 or 14 days posttreatment. Transgene expression was assessed using immunohistochemistry and bioluminescent assays. The highest transfection rate was observed 48 h posttransfection: 79% for HKC, 70% for HFB, and 48% for HaCaT. The eGFP expression was detectable in all groups over 30 days (P>0.05). Cytotoxic effects of the adenoviral vector were observed for HFB after 10 days and HaCaT after 30 days. Reporter gene expression in vivo was significantly higher in burned skin compared with unburned skin (P=0,004). Gene expression decreases from 2 to 7 days with no significant expression after 14 days. This study demonstrates that effective adenoviral-mediated gene transfer of epidermal primary cells and cell-lines is feasible. Ex vivo gene transfer in epithelial cells might have promise for the use in severely burned patients who receive autologous keratinocyte sheets. Transient cutaneous gene delivery in burn wounds using adenoviral vectors causes significant concentrations in the wound tissue for at least 1 week. Based on these findings, we hypothesize that transient cutaneous adenoviral gene delivery of wound healing promoting factors has

Uranium Content in the Geological Samples of Different River Valleys in the Dauki fault Belt of Jaintiapur

International Nuclear Information System (INIS)

Chowdhury, Minhaz ul Islam; Ahmed Monir; Bhuiyan Abu Daiyan; Akon Eunus

1996-01-01

Thirty four geological samples that include six radioactive rock samples, four soil samples, two ooze samples,twelve stream-sediment samples and ten water samples, collected during a field survey in Jaintiapur area of the Dauki fault belt, oriented to the study on probable mobility of uranium either as detritus or in aquatic solution through the region, have been investigated with fluorimeter so as to find out content of uranium in the samples. The investigation aims at determining geochemical significance on uranium occurrence prevailing in the area. In general, the content of uranium in the rock samples lies in average distribution of geochemical interest. However, 194 ppm of uranium content in the gross material of the rock sample, collected from Lalakhal anomaly that records 1500 cps in situ, bears geochemical significance. But, the anomaly being associated with a cross -bedding, the presence of uranium may be inferred as an accumulation in placer sediment transported from a nearby source, Despite absence of in situ radiometric anomalies in the overall geological formations other than the Tipam and the Dupitila Sandstone members as encountered in the traverse of Sari valley , the area, in general, appears to be potential for possible occurrence of uranium. The transported stream sediments ooze and surface run-off water, as collected from the Rangapani, the Sari and even the Nayagang show geochemically significant distribution of uranium. Laboratory analyses of soil samples refers to dispersion of U bearing materials in the soil along the major channels.Uranium content in the surface run-off water of the Sari, the Rangapani and the Nayagang that have originated from the uranium bearing Meghalayan hills strongly support previous inference on mobility of uranium in aquatic solution through the geological formations of the area for possible formation of secondary uranium deposits. Eventually, prevailing geological evidences advocate that the area may be brought

Rapid Simultaneous Amplification and Detection of the MBR/JH Chromosomal Translocation by Fluorescence Melting Curve Analysis

Science.gov (United States)

Bohling, Sandra D.; King, Thomas C.; Wittwer, Carl T.; Elenitoba-Johnson, Kojo S. J.

1999-01-01

Polymerase chain reaction (PCR) amplification and product analysis for the detection of chromosomal translocations, such as the t(14;18), has traditionally been a two-step process. PCR product detection has generally entailed gel electrophoresis and/or hybridization or sequencing for confirmation of assay specificity. Using a microvolume fluorimeter integrated with a thermal cycler and a PCR-compatible double-stranded DNA (dsDNA) binding fluorescent dye (SYBR Green I), we investigated the feasibility of simultaneous thermal amplification and detection of MBR/JH translocation products by fluorescence melting curve analysis. We analyzed DNA from 30 cases of lymphoproliferative disorders comprising 19 cases of previously documented MBR/JH-positive follicle center lymphoma and 11 reactive lymphadenopathies. The samples were coded and analyzed blindly for the presence of MBR/JH translocations by fluorescence melting curve analysis. We also performed dilutional assays using the MBR/JH-positive cell line SUDHL-6. Multiplex PCR for MBR/JH and β-globin was used to simultaneously assess sample adequacy. All (100%) of the 19 cases previously determined to be MBR/JH positive by conventional PCR analysis showed a characteristic sharp decrease in fluorescence at ∼90°C by melting curve analysis after amplification. Fluorescence melting peaks obtained by plotting the negative derivative of fluorescence over temperature (−dF/dT) versus temperature (T) showed melting temperatures (Tm) at 88.85 ± 1.15°C. In addition, multiplex assays using both MBR/JH and β-globin primers yielded easily distinguishable fluorescence melting peaks at ∼90°C and 81.2°C, respectively. Dilutional assays revealed that fluorescence melting curve analysis was more sensitive than conventional PCR and agarose gel electrophoresis with ultraviolet transillumination by as much as 100-fold. Simultaneous amplification and fluorescence melting curve analysis is a simple, reliable, and sensitive method

Development of a biosensor protein bullet as a fluorescent method for fast detection of Escherichia coli in drinking water.

Science.gov (United States)

Gutiérrez-Del-Río, Ignacio; Marín, Laura; Fernández, Javier; Álvarez San Millán, María; Ferrero, Francisco Javier; Valledor, Marta; Campo, Juan Carlos; Cobián, Natalia; Méndez, Ignacio; Lombó, Felipe

2018-01-01

Drinking water can be exposed to different biological contaminants from the source, through the pipelines, until reaching the final consumer or industry. Some of these are pathogenic bacteria and viruses which may cause important gastrointestinal or systemic diseases. The microbiological quality of drinking water relies mainly in monitoring three indicator bacteria of faecal origin, Escherichia coli, Enterococcus faecalis and Clostridium perfringens, which serve as early sentinels of potential health hazards for the population. Here we describe the analysis of three chimeric fluorescent protein bullets as biosensor candidates for fast detection of E. coli in drinking water. Two of the chimeric proteins (based on GFP-hadrurin and GFP-pb5 chimera proteins) failed with respect to specificity and/or sensitivity, but the GFP-colS4 chimera protein was able to carry out specific detection of E. coli in drinking water samples in a procedure encompassing about 8 min for final result and this biosensor protein was able to detect in a linear way between 20 and 103 CFU of this bacterium. Below 20 CFU, the system cannot differentiate presence or absence of the target bacterium. The fluorescence in this biosensor system is provided by the GFP subunit of the chimeric protein, which, in the case of the better performing sensor bullet, GFP-colS4 chimera, is covalently bound to a flexible peptide bridge and to a bacteriocin binding specifically to E. coli cells. Once bound to the target bacteria, the excitation step with 395 nm LED light causes emission of fluorescence from the GFP domain, which is amplified in a photomultiplier tube, and finally this signal is converted into an output voltage which can be associated with a CFU value and these data distributed along mobile phone networks, for example. This method, and the portable fluorimeter which has been developed for it, may contribute to reduce the analysis time for detecting E. coli presence in drinking water.

Synthesis and characterization of lanthanide-based luminescent nanoparticles: toward new bio-labels

International Nuclear Information System (INIS)

Lechevallier, Severine

2010-01-01

This work deals with the development of luminescent nanoparticles (NPs) suitable as bio-labels. Inorganic NPs have been synthesized, in which luminescent ion (Eu 3+ in most of cases) is substituting ions of the oxide host matrix. NPs of Ln(OH)CO 3 :Eu 3+ (Ln = Y or Gd), Ln 2 O 3 :Eu 3+ , Ln 2 O 2 S:Eu 3+ and SiO 2 :Eu 3+ have been characterized by the way of TGA, WAXS-XRD, TEM, IR and photoluminescence (PL) techniques. The controlled precipitation using urea as precipitating agent is the way chosen and optimized to obtain spherical and monodispersed in size (150±15 nm) NPs. These particles of amorphous lanthanide hydroxycarbonate can directly be used as luminescent bio-labels or after their conversion in oxide or oxi-sulfide. For the silica particles, the synthesis by aerosol pyrolysis has been used. The obtained particles are spherical with a main diameter of 350 nm. In a second step, the surface of the inorganic NPs has been modified, in order to graft amino-reactive functions. Several modification ways have been explored: with APTES (aminopropyltriethoxysilane), with TEOS (tetra-ethoxysilane) and then APTES, or with a TEOS/APTES mixture. The same characterizations techniques have been applied to the modified particles, and chemical analysis, solid state NMR and XPS. DLS and ζ-potential of the NPs dispersed in water have also been measured. These analyses have been completed by the evaluation of the number of accessible amine functions by coupling with FITC (fluorescein isothiocyanate). The direct modification of oxides (Y 2 O 3 or Gd 2 O 3 ) with APTES is the best way, and an homogenous layer of 1 nm with a high number of accessible amine can be graft. Finally, to move toward luminescent bio-labelling in biological medium, the luminescence of the NPs has been observed and analyzed using: - A spectro-fluorimeter, after their dispersion in water; - A fluorescence microscope, on glass slides, under broad band excitation; - A confocal microscope, under laser

Application of fission track technique for estimation of uranium concentration in drinking waters of Punjab

International Nuclear Information System (INIS)

Prabhu, S.P.; Raj, Sanu S.; Sawant, P.D.; Kumar, Ajay; Sarkar, P.K.; Tripathi, R.M.

2010-01-01

Full text: Drinking water samples were collected from four different districts, namely Bhatinda, Mansa, Faridkot and Firozpur, of Punjab for ascertaining the U(nat.) concentrations. The samples were collected from bore wells, hand pumps, tube wells and treated municipal water supply. All these samples (235 nos.) collected were preserved and processed by following the international standard protocol and analyzed by Laser Fluorimetry. Results of analysis by laser fluorimetry have been already reported. To ensure accuracy of the data obtained by laser fluorimetry, few samples (10 nos) from each district were analyzed by alpha spectrometry as well as by fission track analysis (FTA) technique. FTA in solution media for uranium has been already standardized in Bioassay laboratory of Health Physics Division. Few of drinking water sample was directly transferred to polythene tube sealed at one end. Lexan detector with proper identification mark was immersed in the samples and the other open end of the tube was also heat-sealed. Two tubes containing samples and one containing uranium standard (80 ppb) were irradiated in the Pneumatic Carrier Facility (PCF) of DHRUVA reactor. The Lexan detectors were then chemically etched and tracks were counted under an optical microscope at 400X magnification. Concentration of uranium in sample was determined by comparison technique. Quality assurance was carried out by replicate analysis and by analysis of standard reference materials. Uranium concentration in these samples ranged from 3.2 to 60.5 ppb with an average of 28.8 ppb. A t-test analysis for paired data was done to compare the results obtained by FTA and those obtained by laser fluorimeter. The calculated value for t is -1.19, which is greater than the tabulated value of t for 40 observations (-2.02 at 95% confidence level). This shows that the results of the measurements carried out by the FTA and laser fluorimetry are not significantly different. The preliminary studies

Real-time monitoring of genetically modified Chlamydomonas reinhardtii during the Foton M3 space mission

Science.gov (United States)

Lambreva, M.; Rea, G.; Antonacci, A.; Serafini, A.; Damasso, M.; Pastorelli, S.; Margonelli, A.; Johanningmeier, U.; Bertalan, I.; Pezzotti, G.; Giardi, M. T.

2008-09-01

. We analysed the hourly changes and the daily light/dark trend in the maximum quantum yield of PSII photochemistry, Fv/Fm (Fig.2). Some physiological parameters that characterize the post-flight effect on algae viability and photosynthetic performance were also determined. The dose and particle flux during Foton-M3 flight were monitored in real time by the active spectrum-dosimeter Liulin- Photo, mounted on the top of Photo-II fluorimeter (Fig.2). Liulin-Photo measurements provided information on the amount of the energy released on the samples and the quality of the incident ionizing radiation [3]. The space flight results in relationship with the ground control simulation are discussed.

Tracing river runoff and DOC over the East Siberian Shelf using in situ CDOM measurements

Science.gov (United States)

Pugach, Svetlana; Semiletov, Igor; Pipko, Irina

2010-05-01

The Great Siberian Rivers integrate meteorological and hydrological changes in their watersheds and play a significant role in the physical and biogeochemical regime of the Arctic Ocean through transport of fresh water (FW) and carbon into the sea. Since 1994, the Laboratory of Arctic Research POI in cooperation with the IARC UAF investigate the fresh water and carbon fluxes in the Siberian Arctic land-shelf system with the special emphasize in the East Siberian Arctic shelf (ESAS) which represents the widest and shallowest continental shelf in the World Ocean, yet it is still poorly explored. The East Siberian Sea is influenced by water exchange from the eastern Laptev Sea (where local shelf waters are diluted mostly by Lena River discharge) and by inflow of Pacific waters from the Chukchi Sea. This region is characterized by the highest rate of coastal erosion and significant volume of the riverine discharge and exhibits the largest gradients in all oceanographic parameters observed for the entire Arctic Ocean. Here we demonstrate a connection among Chromophoric (or Colored) Dissolved Organic Matter (CDOM) which represents the colored fraction of Dissolved Organic Carbon (DOC), salinity, and pCO2. Our data have documented strong linear correlations between salinity and CDOM in the near shore zone strongly influenced by riverine runoff. Correlation coefficient between CDOM and salinity in surface waters was equal to -0.94, -0.94 and -0.95 for surface water stations in September of 2003, 2004, and 2005, respectively. Combined analysis of CDOM and DOC data demonstrated a high degree of correlation between these parameters (r=0.96). Such close connection between these characteristics of waters in this region makes it possible to restore the distribution of DOC according to our original CDOM data of the profiling systems, such as CTD-Seabird equipped by WETStar CDOM fluorimeter. It is shown that the CDOM can be used as a conservative tracer to follow the transport and

Origin, conditions and processes of sandstone reservoir diagenetic silicification in the North Sea; Origine, conditions et processus de la silicification diagenetique de reservoir greseux en Mer du Nord

Energy Technology Data Exchange (ETDEWEB)

Blanchet, A.

2002-02-01

The petroleum reservoir qualities of sandstones are partially reduce by the presence of diagenetic quartz which occlude the cement porosity. Two sandstones reservoirs have been studied to understand the process and origin of silicification. The Brent sandstones have been sampled in the Alwyn north and Dunbar fields (North Sea) and the Franklin sandstones in the Elgin and Glenelg fields (North Sea). Fluid inclusions are often used to constrain quartz precipitation temperature. However their possible stretching with burial is still a problem. Fluid inclusion (FI) petrographic and micro-thermometric study does not show any evidence of stretching: no correlation between size, shape and homogenization temperature (Th) has been observed. Large Th range, even for FI in a single detrital grain overgrowth boundary, cannot been explained by temperature reset. The fluorescence of hydrocarbon inclusions has been analysed by micro-spectro-fluorimeter (MSF) and interpreted as density after calibration using known crude oils. Hydrocarbon inclusions show large density range for FI trapped along a single detrital grain overgrowth boundary. Raman spectroscopy reveals that aqueous inclusions are not saturated with methane. Moreover, a linear relation between methane content and Th implies pressure corrections reducing the range of trapping temperatures. The similarity of Th and MSF data between FI located along detrital grain-overgrowth boundary and within the overgrowth of a single grain indicates that FI located along the boundary are trapped after the precipitation of the first overgrowth zone. These inclusions cannot then be used to represent the beginning of the silicification. The diagenetic sequence and aluminium content in the quartz overgrowth allow to establish the origin of silica for each overgrowth zone defined in cathodoluminescence. High aluminium contents (up to 1375 ppm) are linked to feldspar dissolution which can be induced by acid meteoric water or organic acids

Evaluation of Low-Drift Nozzles in Agrochemical Applications in Orchards Evaluación de Boquillas Antideriva en Aplicaciones de Agroquímicos en Huertos Frutales

Directory of Open Access Journals (Sweden)

Sergio Behmer

2010-09-01

Full Text Available Fruit production demands several phytosanitary treatments per year. The efficiency of these treatments is affected by product loss because of drift, a situation that is aggravated by frequent high winds and the presence of a substantial rural population. The objective of this study was to evaluate the distribution and to quantify product losses in fruit orchard applications. Fluorescent tracers with air-blast sprayers were applied, using air-induction hollow cone nozzles, compared to conventional hollow cone nozzles. To evaluate drift, pipes cleaner were placed on columns behind the adjacent row to the treated one, and for the losses under the tree, Petri dishes were located below the treated row; for the distribution in the tree, leaves were collected from the canopy. The deposits were quantified with a fluorimeter, and the surface of the leaves with an area optical meter. Both nozzles produced equal quantity of deposits in the canopy with a minor quantity in the top part. The low drift sprinklers obtained 75% less drift than the standard nozzles; however, the losses under the tree were 50% higher. To conclude, the utilization of low drift nozzles in fruit orchards is an alternative to mitigate pollution of the environment.La producción frutícola exige varios tratamientos fitosanitarios anuales, cuya eficiencia se ve afectada por la deriva. Esta situación se agrava por la alta frecuencia de vientos y por la existencia de una importante población rural. El objetivo del trabajo fue evaluar la distribución en el árbol y cuantificar las pérdidas de producto en aplicaciones en huertos frutales. Se aplicó un trazador fluorimétrico utilizando un pulverizador hidroneumático en dos versiones: A con boquillas de cono con inducción de aire, y B con boquillas de cono estándar. Para evaluar la deriva se colocaron limpiadores de pipas en columnas ubicadas detrás de la fila adyacente a >la tratada. Las pérdidas debajo del árbol se recolectaron

Use of radon as tracer for identification of aquifer discharge along the Matinha stream, MG, Brazil; O uso do radonio como tracador para identificacao de descarga de aquiferos ao longo do corrego da Matinha, MG

Energy Technology Data Exchange (ETDEWEB)

Chagas, Claudio Jose

2017-11-01

activities. The fluorimeter used to recover the mass of injected fluorescence tracer was the GGUN FL-30 connected to a notebook so that it could follow the tracer cloud passage. The gamma radiation measurements were fitted with AT6101C detector installed inside a backpack so that they could be scanned along the thalweg and along the banks of the stream. The results showed the presence of significant amounts of the radioisotope in several sections of Matinha stream, which indicates the interconnection between the groundwater and the surface waters. This interconnection with aquifers justifies the increase of the flows found between upstream and downstream of the sections with larger activities of {sup 222}Rn, since there were no other superficial contributions along the section studied. The gamma radiation found denotes proximity to the base rock along with the minerals brought by the groundwater and sedimented on the flood surfaces. (author)

Use of radon as tracer for identification of aquifer discharge along the Matinha stream, MG, Brazil

International Nuclear Information System (INIS)

Chagas, Claudio Jose

2017-01-01

Within the framework of 'Nuclear Techniques' is the use of natural and artificial isotopes as tracers for the study of the environment. The use of natural tracers in hydrology is a very useful tool that has been used in several scenarios. One such tracer is the isotope of radon, "2"2"2Rn, a noble gas from natural sources as a consequence of alpha decay of "2"2"6Ra. Radon can be found, to a greater or lesser degree, in all groundwater, as well as in soils and rocks from where it exudes into the atmosphere through spaces arising from fissures and or nanopores present. It can emerge into the water bodies by the leaching of water or other liquids through these interstices. In this research, the "2"2"2Rn gas diluted naturally by the leaching of the water that infiltrated the soils and rocks of the aquifers was used as a tracer in order to identify sections of discharge of these aquifers along the Stream of Matinha. The study area is about 70 km from Belo Horizonte and is in the city of Itauna, MG. The Matinha stream is about 2,200 m in length and is part of the network of tributaries of the water reservoir called Serra Azul, in the Juatuba River Basin, MG. The results presented express the studies that occurred between the second half of 2014 and the first half of 2016, in ten campaigns for water sampling, flow and natural gamma radiation measurements. Water samples were collected near the bottom and very close to the thalweg in the watercourse using syringe and or peristaltic suction pump at low flow to avoid gas leakage. The "2"2"2Rn analyzes were performed in the field during the collections and in the laboratory of the CDTN - Centro de Desenvolvimento da Tecnologia Nuclear, all of which were carried out with the radon detector RAD 7. The flow measurements counted on the use of the fluorescent tracer's technique: rhodamine and fluorescein were used for injection at points upstream and downstream of sections with higher "2"2"2Rn activities. The fluorimeter used to

New Insights into the Functional Behavior of Antibodies as Revealed by Binding Studies on an Anti-Uranium Monoclonal Antibody

International Nuclear Information System (INIS)

Blake, Diane A.; Xia Li; Haini Yu; Blake, Robert C.

2004-01-01

As part of an ongoing effort to develop immunoassays for chelated uranium(VI) on a hand-held flow fluorimeter, an anti-uranium monoclonal antibody designated as 8A11 was fluorescently labeled using two different strategies. When 8A11 was coupled via reactive lysines to either ALEXATM 488 or Cy5TM, the resulting fluorescent antibody conjugate exhibited positive cooperativity in the presence of its antigen, U(VI) chelated with 2,9-dicarboxy-1,10-phenanthroline (U(VI)-DCP). That is, when one of the two binding sites on the covalently modified 8A11 was occupied with bound antigen, the affinity of the remaining site on the antibody for U(VI)-DCP appeared to increase. Unmodified 8A11 bound U(VI)-DCP with the expected hyperbolic dependence on the concentration of antigen, consistent with independent and equal binding of ligand at both sites. Proteolytic cleavage of the fluorescently conjugated 8A11 to produce the fluorescent monovalent Fab fragment yielded an active preparation that now bound U(VI)-DCP with no evidence of positive cooperativity. Although, in principle, any divalent antibody has the potential to exhibit positive cooperativity in its binding interactions with its antigen, very little literature precedent for this type of behavior exists. Native 8A11 was also noncovalently labeled with highly fluorescent ZENONTM reagents. These reagents are fluorescently-labeled Fab fragments of goat anti-mouse antibodies that bind to the Fc portion of 8A11. These high-affinity, monovalent fluorescent reagents permitted the intact 8A11 mouse antibody to be labeled in situ with no covalent modifications. Incubation of the 8A11 with ZENON 647 produced a fluorescent protein complex that showed an 8-fold higher affinity for U(VI)-DCP than did the free 8A11 alone. Again, very few literature precedents exist for this phenomenon, where agents that bind to the Fc portion of an intact antibody change the affinity of the antibody for the antigen at the structurally distant Fab portion

The use of Fluorescence Resonance Energy Transfer (FRET peptidesfor measurement of clinically important proteolytic enzymes

Directory of Open Access Journals (Sweden)

Adriana K. Carmona

2009-09-01

Full Text Available Proteolytic enzymes have a fundamental role in many biological processes and are associated with multiple pathological conditions. Therefore, targeting these enzymes may be important for a better understanding of their function and development of therapeutic inhibitors. Fluorescence Resonance Energy Transfer (FRET peptides are convenient tools for the study of peptidases specificity as they allow monitoring of the reaction on a continuous basis, providing a rapid method for the determination of enzymatic activity. Hydrolysis of a peptide bond between the donor/acceptor pair generates fluorescence that permits the measurement of the activity of nanomolar concentrations of the enzyme. The assays can be performed directly in a cuvette of the fluorimeter or adapted for determinations in a 96-well fluorescence plate reader. The synthesis of FRET peptides containing ortho-aminobenzoic acid (Abz as fluorescent group and 2, 4-dinitrophenyl (Dnp or N-(2, 4-dinitrophenylethylenediamine (EDDnp as quencher was optimized by our group and became an important line of research at the Department of Biophysics of the Federal University of São Paulo. Recently, Abz/Dnp FRET peptide libraries were developed allowing high-throughput screening of peptidases substrate specificity. This review presents the consolidation of our research activities undertaken between 1993 and 2008 on the synthesis of peptides and study of peptidases specificities.As enzimas proteolíticas têm um papel fundamental em muitos processos biológicos e estão associadas a vários estados patológicos. Por isso, o estudo da especificidade das peptidases pode ser importante para uma melhor compreensão da função destas enzimas e para o desenvolvimento de inibidores. Os substratos com supressão intramolecular de fluorescência constituem uma excelente ferramenta, pois permitem o monitoramento da reação de forma contínua, proporcionando um método prático e rápido para a determinação da

Fast neutron biological effects on normal and tumor chromatin

International Nuclear Information System (INIS)

Constantinescu, B.; Bugoi, Roxana; Paunica, Tatiana; Radu, Liliana

1997-01-01

the relative absorbencies (E/E 0 ) obtained for the thermal transitions of chromatin samples versus fast neutron dose, lower values at higher neutron dose, were observed, indicating a damaged chromatin DNA. The treatment with gyrostan accelerates the damage process, but adding thyroxine and D3 vitamin the negative effect is partially recovered. Some conclusions on the role of gyrostan and of thyroxine and D3 vitamin are presented. To study the influence of metal ions on DNA strand breakage induced by fast neutrons, chromatin irradiations with 100, 200 and 300 Gy were performed separately and in presence of CsCl, CuCl 2 and AlCl 3 in a 5 x 10 -2 M concentration. Fluorescence lifetime values for chromatin-ethidium bromide complexes were determined using a Fluorimeter Edinburgh Anal. Instr. FL-900 CD. Greater values denote a damaged chromatin DNA. The presence of metal cations strongly decreases the time of life values, acting as a trap for the water radiolysis products. Our results suggest a combination of fast neutron irradiation with anticancer drug gyrostan for tumor destruction enhancement and the use of metal ions with hormonal thyroxine and D3 vitamin for a normal cells better protection during neutron therapy.(authors)

The Measurement of Integrated Gamma-Ray Doses in the Range 50 Mrad to 5 Mrad Using Phosphate Glass and Lithium Fluoride; Mesure, au Moyen de Verre au Phosphate et de Fluorure de Lithium, de Doses Integrees de Rayons Gamma de 50 Mrad a 5 Mrad; 0418 0417 041c 0414 ; Medicion de Dosis Integradas de Rayos Gamma de 50 Mrad a 5 Mrad por Medio de Vidrio al Fosfato y de Fluoruro de Litio

Energy Technology Data Exchange (ETDEWEB)

Webb, G. A.M. [Central Electricity Generating Board, Berkeley Nuclear Laboratories, Berkeley, Glos. (United Kingdom)

1965-06-15

The radio-photoluminescence and colour-change of phosphate glass and the thermoluminescence of lithium fluoride have been compared as the basis of dosimetry systems over the range of {gamma}-ray doses from 50 mrad to 5 Mrad. The stimulated luminescence of low atomic number, silver-activated phosphate flass was measured on a commercial fluorimeter (Toshiba fluoroglass dosimeter) supplemented by a spectrophotometer to measure the change of optical density for the high-dose region. The effects of various cleaning procedures for the glass were studied. The thermoluminescence of lithium fluoride was measured using a laboratory reader to produce glow curves and then integrating the area under the required peak. The emission spectrum of the thermoluminescence was measured using optical filters, which were also used to minimize the effects due to infra-red radiation from the heating tray. The effects on the tribothermoluminescent peak of optical filters and readout under nitrogen were investigated. A number of performance criteria of the two systems were investigated and compared critically. The range of the glass is very large, from 5 x 10{sup -2} to 5 x 10{sup 6} rad, i. e. a dynamic range of 108, whereas LiF, although it may be used for slightly lower doses, saturates above 10{sup 5} rad. The LiF is almost energy-dependent, but glass shows a maximum departure of seven times the response to Co{sup 60} at 60 keV; this may be corrected by shielding at the expense of raising the lower energy limit. The fading of the glass has been thoroughly investigated; that of LiF, which is not so marked, has been studied to a lesser extent. Annealing and re-use of both glass and LiF is possible and various annealing cycles have been tried for each; the sensitivity for re-use has also been measured. The effects of temperature of irradiation and storage in the range 20 to 100 Degree-Sign C have been studied for each system, as has the evaluation of doses in mixed neutron and {gamma