一种适合14C-尿素呼气试验的环境友好型闪烁液的效果评价

目的评价环境友好型闪烁液应用于14C-尿素呼气试验诊断幽门螺杆菌感染的效果。方法149例患者接受14C-尿素呼气试验,均分别采用环境友好型和传统液体闪烁液进行检测,比较两者结果。结果应用环境友好型闪烁液进行14C-尿素呼气试验诊断幽门螺杆菌感染,与应用传统型闪烁液相比,两者差异无显著性,结果符合率为97.99%。结论环境友好型闪烁液对14C-尿素呼气试验结果无明显负面影响,可替代传统闪烁液。     

A thermostable cocktail for the liquid scintillation counting of heterogeneous media

Considerable analytical errors arise in the liquid scintillation counting of heterogeneous media as a consequence of gel instability. With large sample numbers, a major causative factor of this instability is temperature changes during the counting period. An emulsifier-scintillation cocktail has been designed to provide stable counting conditions for heterogeneous media over a temperature range of 10–30°C, i.e., the wide range of temperature likely to be encountered in liquid scintillation counters lacking sample cooling facilities. A comparison was made with a conventional commercially available emulsifier-scintillator.     

Use of scintillometric quantitation of unscheduled DNA synthesis in isolated rat hepatocytes for the screening of genotoxic agents

The induction of unscheduled DNA synthesis has been considered as a suitable endpoint for the screening of genotoxic agents. Experimentally, unscheduled DNA synthesis is most frequently measured by autoradiography. The purpose of this report was to examine the usefulness of the liquid scintillation counting technique in measuring unscheduled DNA synthesis response in isolated rat hepatocytes. The various liquid scintillation counting-based unscheduled DNA synthesis assay procedures were examined according to the following groupings: (1) procedures based on the acid precipitation of cellular macromolecules, (2) procedures based on isopycnic gradient centrifugation of solubilized cells, (3) procedures based on nuclei isolation in conjunction with other DNA purification methods, and (4) procedures based on the selective retention of hepatocellular DNA. Limited cases in which test chemicals gave positive unscheduled DNA synthesis response in liquid scintillation counting-based assays and negative unscheduled DNA synthesis response in autoradiography-based assays are presented. It is concluded that liquid scintillation counting-based unscheduled DNA synthesis assays represent an appropriate system for inclusion in carcinogenicity and mutagenicity testing programs.Abbreviations 2-AAF 2-acetylaminofluorene - 2-AF 2-aminofluorene - AFB₁ aflatoxin B₁ - ARG autoradiography - DMN dimethylnitrosamine - LSC liquid scintillation counting - MMS methyl methanesulfonate - MNNG N-methyl-N-nitro-N-nitrosoguanidine - 4-NQO 4-nitroquinoline-1-oxide - PCA perchloric acid - TCA trichloroacetic acid - UDS unscheduled DNA synthesis     

Dynamics of Pulsed X-Ray Radiation of a Plasma Micropinch Discharge

Plasma Physics Reports - A complex of diagnostic equipment has been created and a measurement procedure has been developed for a multi-channel scintillation X-ray spectrometer with nanosecond time...     

An effielent seintillation radioassay system and sensitive protein assay for submilligram quantities of subcellular membranes

A simple combination of commercially available vials has been devised to reduce both the system volume (2–4 ml) and the consumption of scintillation chemicals for radioassay. Counting efficiency is improved by adding a transparent liquid between the vials. The estimation of protein in the samples either before or after admixture with scintillation fluid is possible with a simple application of the trinitrobenzenesulfonic acid method, following removal of nonprotein trinitrophenylated material by a butanol extraction.     

A novel cell-based scintillation proximity assay for studying protein function and activity in vitro using membrane-soluble scintillants

Here we describe for the first time a cell-based scintillation proximity assay using membrane soluble scintillants (MSS). MSS have a scintillant "head" group (2,5-diphenyloxazole) attached to a lipophilic "tail." MSS do not scintillate in an aqueous environment in the presence of a radioactive source: however, in a non-aqueous environment, such as a lipid bilayer (e.g., liposome or cell membrane), scintillation does occur. MSS can be incorporated into liposomes. When these MSS-containing liposomes are fused with the plasma membranes of cells in culture the MSS are incorporated into the cell membrane. Radiolabelled molecules in close proximity to the cell membrane will then elicit a scintillation signal. This system has been used to successfully monitor [(14)C]methionine uptake in HeLa cells and may be used in radiochemical and radioligand binding assays either in vivo or on microsomal preparations obtained from tissues. This new scintillation proximity technology could be readily adapted for high-throughput screening.     

A rapid method to determine the mammalian cell cycle

A method was developed for determining the duration of the mammalian cell cycle and each of its major phases, mitosis, G1, DNA synthetic period, and G2. Mitotic time was determined by assessment of the mitotic index at intervals after cells collected in mitosis and stored at 4 °C were reincubated at 37 °C. The duration of the three remaining phases was derived from a graphic representation of the uptake of ³H-thymidine by a synchronous population of cells grown directly in scintillation vials. The scintillation counting method for determination of these parameters is advantageous over methods using autoradiography in that the investigator's bias in scoring cells is eliminated. Complex mathematical interpretations are unnecessary, and the data obtained from the scintillation counter are readily processed. Results from scintillation counting and autoradiographic methods are shown to be comparable.     

Improvements in and Environmental Applications of Double-Vial Radiorespirometry for the Study of Microbial Mineralization

Several variations in the scintillation mixture and the filter paper arrangements for double-vial radiorespirometry were compared. Improved efficiencies (44%) and shorter response times were found by adding wetting agents and methanolic NaOH to the scintillation mixture in the filter paper. The scintillation chemicals used did not contain dioxane and were found to be nontoxic to the test microbiota in this system. Covering the inner reaction vial with aluminum foil minimized the reduction in counting efficiency when testing colored or dense environmental samples. Mineralization rates were determined with ¹⁴C-labeled glucose, acetate, and glutamate and [¹⁴C]cellulose- and [¹⁴C]lignin-labeled lignocellulose for composting cow manure, forest soil, and arctic lake sediment microbiota. This improved method can be used in a variety of procedures involving the measurement of microbial mineralizations of organic compounds. Since no liquid scintillation cocktail is used for counting, the radioactive wastes are aqueous or can be incinerated, making disposal easy.     

In silico investigation of factors affecting the MV imaging performance of a novel water-equivalent EPID

PurposeA Geant4 model of a novel, water-equivalent electronic portal imaging device (EPID) prototype for radiotherapy imaging and dosimetry utilising an array of plastic scintillating fibres (PSFs) has been developed. Monte Carlo (MC) simulations were performed to quantify the PSF-EPID imaging performance and to investigate design aspects affecting performance for optimisation.MethodsUsing the Geant4 model, the PSF-EPID’s imaging performance for 6 MV photon beams was quantified in terms of its modulation transfer function (MTF), noise power spectrum (NPS) and detective quantum efficiency (DQE). Model parameters, including fibre dimensions, optical cladding reflectivity and scintillation yield, were varied to investigate impact on imaging performance.ResultsThe MC-calculated DQE(0) for the reference PSF-EPID geometry employing 30 mm fibres was approximately nine times greater than values reported for commercial EPIDs. When using 10 mm long fibres, the PSF-EPID DQE(0) was still approximately three times greater than that of a commercial EPID. Increased fibre length, cladding reflectivity and scintillation yield produced the greatest decreases in NPS and increases in DQE.ConclusionsThe potential to develop an optimised next-generation water-equivalent EPID with MV imaging performance at least comparable to commercial EPIDs has been demonstrated. Factors most important for optimising prototype design include fibre length, cladding reflectivity and scintillation yield.     

A rapid micromethod for the determination of nitrogen and phosphate in biological material

Two types of small containers for the liquid scintillation counting of 0.15–2 ml of aqueous sample in emulsifying agents have been investigated. Counting efficiency and reproducibility were found to be comparable with those found with more orthodox methods, but the effective cost is less. The systems also provide an economical way of using expensive reagents.     

Replication of the R6K plasmid during the Escherichia coli cell cycle.

The cell-cycle replication pattern of the R6K plasmid has been investigated by using the membrane-elution technique to produce cells labelled at different times during the division cycle and scintillation counting for quantitative analysis of radioactive plasmid DNA. The high-copy plasmid R6K replicates exponentially in a cell-cycle-independent manner. A mini-R6K plasmid deleted for the ori alpha origin of replication also replicates, exponentially in a cell-cycle-independent manner.     

Scintillation proximity assay for measuring uptake by the human drug transporters hOCT1, hOAT3, and hOATP1B1

Increasing evidence suggests a key role of transport proteins in the pharmacokinetics of drugs. Within the solute carrier (SLC) family, various organic cation transporters (OCTs), organic anion transporters (OATs), and organic anion transporting polypeptides (OATPs) that interact with drug molecules have been identified. Traditionally, cellular uptake assays require multiple steps and provide low experimental throughput. We here demonstrate the use of a scintillation proximity approach to detect substrate uptake by human drug transporters in real time. HEK293 cells stably transfected with hOCT1, hOATP1B1, or hOAT3 were grown directly in Cytostar-T scintillating microplates. Confluent cell monolayers were incubated with 14C- or 3H-labeled transporter substrates. Cellular uptake brings the radioisotopes into proximity with the scintillation plate base. The resulting light emission signals were recorded on-line in a microplate scintillation counter. Results show time- and concentration-dependent uptake of 14C-tetraethylammonium, 3H-methylphenylpyridinium (HEK-hOCT1), 3H-estradiol-17beta-D-glucuronide (HEK-hOATP1B1), and 3H-estrone-3-sulfate (HEK-hOAT3), while no respective uptake was detected in empty vector-transfected cells. Km of 14C-tetraethylammonium and 3H-estrone-3-sulfate uptake and hOAT3 inhibition by ibuprofen and furosemide were similar to conventional dish uptake studies. The scintillation proximity approach is high throughput, amenable to automation and allows for identification of SLC transporter substrates and inhibitors in a convenient and reliable fashion, suggesting its broad applicability in drug discovery.     

Branched-chain-amino-acid aminotransferase assay using radioisotopes

A method for the assay of the activity of branched-chain-amino-acid aminotransferase from Escherichia coli has been developed. Radioactive isoleucine is used, the radioactive oxoacid formed in the enzymic reaction is converted to its p-nitrophenylhydrazone, and the hydrazone is extracted into toluene based scintillation liquid. The small reaction tubes containing the toluene layer and the reaction mixture as a water layer are placed into liquid scintillation vials and counted for radioactivity. The radioactive amino acid remaining in the water layer causes only a rather low background.     

Scintigrams of the thyroid gland; the diagnosis of morphologic abnormalities with I131

Functioning thyroid tissue containing sufficient radioiodine can be visualized by scanning the gland with a directional scintillation counter.(4) This visual representation of the gland is called a "scintigram." Scintigrams have been invaluable in the detection and study of both "toxic" and non-functioning nodules, diffuse enlargement in hyperthyroidism and the subsequent reduction in gland size after treatment, carcinoma, and aberrant thyroid tissue.     

Chemiluminescence in a toluene-fenoxol scintillation system in the radioreceptor method of researching biological membranes

The scintillation toluene-fenoxol 8/10 system with ammonia (0.01%) is suggested for radioactive counting in radioreceptor biomembrane assay. It is shown that chemiluminescence in this system is defined mainly by membrane components presence. The remotion of filter from scintillator after labelled molecules elution and methanol (3%) addition to scintillation mixture are suggested to reduce the chemiluminescence level.     

Liquid scintillation counting of 14C-labeled hemin and its application to the biosynthesis of heme in bone marrow

A rapid procedure is described for the liquid scintillation counting of ¹⁴C-labeled hemin isolated after incubation of bone marrow with radioactively labeled glycine-2-¹⁴C. The method has been applied for studies on the biosynthesis of heme in bone marrow of several animal species.     

A generic high-throughput screening assay for kinases: protein kinase a as an example

A generic high-throughput screening assay based on the scintillation proximity assay technology has been developed for protein kinases. In this assay, the biotinylated (33)P-peptide product is captured onto polylysine Ysi bead via avidin. The scintillation signal measuring the product formation increases linearly with avidin concentration due to effective capture of the product on the bead surface via strong coulombic interactions. This novel assay has been optimized and validated in 384-well microplates. In a pilot screen, a signal-to-noise ratio of 5- to 9-fold and a Z' factor ranging from 0.6 to 0.8 were observed, demonstrating the suitability of this assay for high-throughput screening of random chemical libraries for kinase inhibitors.     

A device for the liberation and determination of 14CO2

A simple closed system for the serial determination of 14CO2 in small volumes of fluid samples, is described. The device consists of commercially available scintillation vials and silicone tube seals. 14CO2 is selectively liberated by citric acid and absorbed in a scintillation vial by Hyamine. Experiments on the effect of dichloroacetate on pyruvate dehydrogenase activity in rat hindlimbs perfused with [1-14C]pyruvate demonstrate the applicability of the method.     

A FlashPlate assay for the identification of PARP-1 inhibitors

A novel FlashPlate scintillation proximity assay has been developed for the high-throughput screening (HTS) of large compound libraries to identify inhibitors of poly(ADP-ribose) polymerase-1 (PARP-1), an important enzyme involved in DNA repair. The assay was originally developed for the 96-well FlashPlate but is easily transferred to a 384-well format. Moreover, the authors demonstrate that the assay is sufficiently sensitive to determine accurate IC(50) values and adaptable for kinetic evaluation of lead molecules. The mechanism of action of the assay requires the binding of PARP-1 to a double-stranded DNA oligonucleotide leading to the active enzyme. Using NAD(+) and (3)H-NAD(+) as substrate, activated PARP-1 synthesizes labeled poly(ADP-ribose) chains. Once the reaction is stopped, ADP-ribose polymers are brought into proximity with the pretreated FlashPlate walls, resulting in signal amplification. This signal is then detected by a TopCount scintillation plate reader. The developed assay is a robust and reproducible method of screening for PARP-1 inhibitors that is low maintenance and cost-effective and can easily be automated.     

Liquid scintillation counting of radioactive monomeric and polymeric carbohydrates on paper chromatograms

A simple, improved scintillation counting procedure was developed for the assay of radioactive mono- and polysaccharides on paper chromatograms. Segments of chromatograms are placed in scintillation vials and soaked in water to completely elute the carbohydrate before addition of Aquasol, a xylene-based scintillation fluid. The resulting water-Aquasol solution is counted in a liquid scintillation counter. Evaluation of numerous experimental variables revealed optimal conditions for complete elution of mono- and polysaccharides with water before counting in Aquasol.The water elution-Aquasol procedure allows water-soluble substances (¹⁴C- and ³H-labeled) on paper to be assayed with increased accuracy and sensitivity (three- to fivefold improvement in counting efficiency of tritiated samples). The simplicity of the procedure allows entire radiochromatograms to be assayed readily.