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Reducing the effect of chemiluminescence on the efficiency of extracting 14C-neomycin from fortified kidney tissue as measured by a liquid scintillation system.

Extraction efficiency was monitored, using 14C-neomycin and liquid scintillation counting, during development of a procedure to isolate neomycin from fortified kidney tissue. Chemiluminescence caused 119% false positive recovery at 0.16 ppm neomycin (1260 disintegrations/min (dpm) activity). However, the contribution of chemiluminescence to false positive recovery was negligible above 8 ppm neomycin (63,000 dpm activity). To reduce chemiluminescence, fortified kidney tissue was extracted with saline by homogenization, mixed with Protosol at 70 degrees C for 1 h, cooled 5 min at 4 degrees C and neutralized with acetic acid. The digest was mixed with Aquasol and incubated 1 h at 47 degrees C, followed by equilibration for 2 h in the counter. The procedure allows reliable monitoring of extraction efficiency down to about 407 dpm activity from 14C-neomycin (0.05 ppm) with maximum chemiluminescence of about 4%.

Animals↗

Serum immunoassay of a small cell lung cancer associated ganglioside: development of a sensitive scintillation proximity assay.

We here report an enzyme linked immunosorbent assay (ELISA) and a scintillation proximity assay (SPA) for detection of the ganglioside FucGM1 in sera from small cell lung cancer (SCLC) patients. The SPA was more sensitive and reproducible than the ELISA. In this assay, monoclonal antibodies specific for FucGM1 were bound to SPA particles and incubated with labelled FucGM1 and 100 microliters test-serum overnight, and counted in a beta-counter. The sensitivity was 0.2 ng. Seven out of twenty sera from SCLC patients were positive, whereas none of twenty sera from healthy individuals were positive for FucGM1. The SPA was more sensitive than the previously reported HPTLC as well as a direct ELISA.

Carbohydrate Sequence↗

A 4pi(LS)beta-gamma coincidence system using a TDCR apparatus in the beta-channel.

This article is an overview of the capabilities of a 4 pi(LS)beta-gamma coincidence counting system based on a liquid scintillation detector in the beta-channel. Equipped with a three-photomultiplier apparatus designed for the triple to double coincidence ratio (TDCR) method, the 4 pi(LS)beta-gamma coincidence system developed at LNHB combines the two techniques. The 4 pi(LS)beta-gamma counting setup records the different types of scintillation events between photomultipliers (double and triple coincidences) and the associated gamma-spectra according to a bi-dimensional configuration. When measuring the activity of a (54)Mn solution, the extrapolation method is carried out by defocusing the photomultipliers; this procedure allows the measurement of gamma-ray sensitivity of the scintillation cocktail in the 835 keV energy region. In addition, in the framework of a (65)Zn standardization, 4 pi(LS)beta-gamma measurements are compared with standard 4 pi(PC)beta-gamma anticoincidence counting using a proportional counter in the beta-channel.

Journal Article↗

An external-sample liquid scintillation counting for 75Se and its application to selenoprotein detection.

An external-sample liquid scintillation (LS) counting for the gamma emitter 75Se has been developed. An expressly designed well-type LS vial and a 2,5-diphenyoxazole-1,4-bis(5-phenyl-2-oxazoyl)-benzene-xylene solution containing 35% tertrabutylzinn allow 75Se to be counted in a standard LS counter with counting efficiency up to 43.2%, much higher than that of conventional LS counting method. This external sample LS has a good count rate linearity and exhibits low background count rates. After in vivo labeling with [75Se]selenite, 75Se distributions and the Se-containing proteins present in tissues of male rat were investigated by means of sodium dodecyl sulfate-polyacrylamide gel electrophoresis, external-sample LS and gamma-detector. Eight Se-containing proteins or protein subunits were detected to be Se-containing proteins or protein subunits in arterial wall, and their apparent molecular masses (Mr) were 76.4, 67.0, 57.4, 30.3, 25.4, 22.7, 21.7, and 15.1 kDa, respectively. In addition, eight 75Se-labeled proteins (Mr: 66.8, 57.0, 43.1, 30.0, 24.8, 19.8, 18.0, and 14.8 kDa) were found in brain homogenates, and nine 75Se-labeled proteins (Mr: 117.0, 78.0, 66.6, 57.2, 43.0, 38.1, 25.0, 20.1, and 18.0 kDa) were detected in testis homogenates. Some of them should be new biologically important selenoproteins that have not been identified so far.

Animals↗

Cadmium uptake and distribution in mouse embryos following maternal exposure during the organogenic period: a scintillation and autoradiographic study.

Pregnant C57BL/6J mice were given a single intraperitoneal injection of either a teratogenic (2,400 micrograms Cd/kg) an intermediate (40 micrograms Cd/kg) or a trace dose (0.66 micrograms Cd/kg) of cadmium chloride (CdCl2) on day 9 of gestation; each dose contained 109CdCl2 (20 microCi). The mice were euthanized at various times after Cd exposure, and the amount of Cd in the embryos was determined in a gamma counter. For all three doses a low, but similar, percentage of the dose rapidly entered the embryos; levels then decreased during the next 11 hours, only to rise again by 24 hours and continue to rise for the remainder of the pregnancy. When the Cd content was related to embryonic weight the Cd concentration was at its highest level after 1 hour and then decreased rapidly and continued to decrease for the rest of pregnancy. Autoradiographic studies showed that the teratogenic dose of Cd entered all tissues of the embryos but was particularly localized in cells of the neural tube, limb buds, and gut. By 12 hours, cell damage was seen in the embryos but the affected cells were not necessarily the heavily labelled cells. In embryos exposed to the nonteratogenic dose the highest Cd accumulation was seen in the embryonic gut and limb bud ectoderm but all tissues showed a low level of labelling.

Animals↗

Monoclonal antibodies for diagnosis and potential therapy of small cell lung cancer--the ganglioside antigen fucosyl-GM1.

Lung carcinomas represent a highly heterogenous group of tumors, which includes small cell lung cancer (SCLC). The ganglioside fucosyl-GM1 (FucGM1) was originally identified as a highly selective marker for SCLC. A number of monoclonal antibodies against FucGM1 have been produced against the purified ganglioside, and their specificity validated. With monoclonal antibodies, FucGM1 has been detected in tissues as well as in serum samples from SCLC patients. A sensitive, quantitative assay method for FucGM1 was developed based on scintillation proximity immunoassay (SPA): A specific monoclonal antibody against FucGM1 is bound to immunosorbent particles that contain a fluor. When radiolabelled FucGM1 binds to these particles, the fluor is excited, and the photons emitted can be measured directly in a beta-counter. This sensitive assay for FucGM1 has several potential diagnostic applications: differential diagnosis of SCLC, development of serum assays for diagnosis and monitoring of disease progression, and monitoring of patient response to therapy. The expression of FucGM1 in SCLC cells is heterogeneous, and may depend on the differentiated state of tumor cells. However, monoclonal antibodies specific for FucGM1 may have important future applications for radioimmunoimaging as well as in immunotherapy for SCLC.

Antibodies, Monoclonal↗

Quality control of gamma counters. I. Experimental evaluation of sources of error.

Errors of counting rate as large as 2 percent may occur through variations in instrumental parameters during the measurement of I-125 activity using a gamma counter. Much larger errors result from failure to maintain constant sample geometry; in particular, the slope of calibration curves depends on sample volume.

Quality Control↗

Lung retention of Pu following inhalation of PuO2 in rats measured using a whole body counter.

A lung retention function on the amount of PuO2 in rats was determined. Five rats were exposed to polydisperse aerosols of PuO2 having a size of 0.47 micron activity median aerodynamic diameter. The initial lung burden was between 1990 Bq and 2960 Bq. Instead of serial-sacrifice study, in vivo counting of low energy L X-rays with thin NaI(Tl) scintillation detectors was used to follow the lung retention of Pu at various intervals up to 468 days after inhalation. The calibration of this counting system was made by measuring lung activity of rats sacrificed for other experimental purposes. It was confirmed that the potential skin contamination and the Pu translocated to the other organs was not counted at the present counting geometry. Our results showed that 77% of PuO2 deposited deep in the lung was cleared with a half-time of 53 days, whereas the residual 23% stayed there for a longer period (a half-time of 795 days).

Administration, Inhalation↗

Intraoperative detection of somatostatin-receptor-positive neuroendocrine tumours using indium-111-labelled DTPA-D-Phe1-octreotide.

After injection of 111In-labelled DTPA-D-Phe1-octreotide, intraoperative tumour localisation was performed using a scintillation detector in 23 patients with neuroendocrine tumours. Count rates from suspect tumour lesions and adjacent normal tissue were expressed as a ratio before (Rin situ) and after (Rex vivo) excision. 111In activity concentration ratios of tumour tissue to blood (T/B) were determined in a gamma counter. In patients with midgut carcinoids, (all scintigraphy positive), false Rin situ recordings were found in 4/29 macroscopically identified tumours. T/B ratios were all high (27-650). In patients with medullary thyroid carcinomas (eight out of ten scintigraphy positive), misleading Rin situ results were found in 4/37 macroscopically identified tumours. T/B ratios were lower (3-39) than those seen in midgut carcinoids. Two out of four patients with endocrine pancreatic tumours had positive scintigraphy, reliable intraoperative measurements and very high T/B ratios (910-1500). One patient with a gastric carcinoid had correct measurements in situ and ex vivo with high T/B ratios (71-210). In situ measurements added little information to preoperative scintigraphy and surgical findings using the present detection system. Rex vivo measurements were more reliable. The very high T/B ratios seen in midgut carcinoids and some endocrine pancreatic tumours would be favourable for future radiation therapy via somatostatin receptors.

Carcinoid Tumor↗

Evaluation of radiation monitor effectiveness for the detection of 85Kr.

As part of the preparations for the purging of TMI Unit-2, the 85Kr sensitivity of 12 radiation detector systems or system combinations was determined. Eleven of these were evaluated using a cube-shaped polyethylene-walled room containing a volume of 5.6 m3 (200 ft3). Krypton-85 gas was added to produce a concentration of 6.7 x 10(-6) muCi/ml in the test room. It was found that none of the ion chambers and scintillation detector systems were able to detect this concentration of 85Kr. Detectors employing thin window GM pancake probes were found to be sensitive enough to monitor this gas down to the unrestrictive area maximum permissible concentration level (MPC) of 3 x 10(-5) muCi/ml, while a large window gas flow proportional counter was found to be sensitive enough to monitor down to about 09.1 MPC. At the end of this experiment, 2.3 m3 (80 ft3) of the gas in the test room was pumped into a compressed air cylinder (scuba bottle) and was used to calibrate The Pennsylvania State University (PSU) Noble Gas Monitor. The sensitivity of this system, which employs gas compression and Ge(Li) spectroscopy, was demonstrated to be between 0.1 and 0.03 times MPC, depending on the counting time employed.

Air Pollutants, Radioactive↗

Evaluation in mice of some iodinated melanoma imaging agents using cryosectioning and multi-wire proportional counting.

AMBIS 4000, a multi-wire proportional counter, was calibrated for iodine-125 measurements. The detector displayed a linear response over a wide dynamic range. Using whole-body mice cryosections, a linear relationship could be established between count rate per area (cpm/mm2) measured with the AMBIS 4000 detector and the count rate per gram (dpm/g) determined with an NaI(Tl) scintillation detector. A calibration curve could, thus be constructed. This new method allowed direct visual and quantitative evaluation of the biodistribution of a short series of 125I-labelled benzamides in melanoma-bearing mice. All the compounds studied showed good tumoral targeting ability. For one of them, ortho-N-(2-diethylaminoethyl)-4-iodobenzamide, liver and lung uptake decreased rapidly after dosing, making it a suitable tracer for scintigraphic detection of malignant melanoma.

Animals↗

PRESCILA: a new, lightweight neutron rem meter.

Conventional neutron rem meters currently in use are based on 1960's technology that relies on a large neutron moderator assembly surrounding a thermal detector to achieve a rem-like response function over a limited energy range. Such rem meters present an ergonomic challenge, being heavy and bulky, and have caused injuries during radiation protection surveys. Another defect of traditional rem meters is a poor high-energy response above 10 MeV, which makes them unsuitable for applications at high-energy accelerator facilities. Proton Recoil Scintillator-Los Alamos (PRESCILA) was developed as a low-weight (2 kg) alternative capable of extended energy response, high sensitivity, and moderate gamma rejection. An array of ZnS(Ag) based scintillators is located inside and around a Lucite light guide, which couples the scintillation light to a sideview bialkali photomultiplier tube. The use of both fast and thermal scintillators allows the energy response function to be optimized for a wide range of operational spectra. The light guide and the borated polyethylene frame provide moderation for the thermal scintillator element. The scintillators represent greatly improved versions of the Hornyak and Stedman designs from the 1950's, and were developed in collaboration with Eljen Technology. The inherent pulse height advantage of proton recoils over electron tracks in the phosphor grains eliminates the need for pulse shape discrimination and makes it possible to use the PRESCILA probe with standard pulse height discrimination provided by off-the-shelf health physics counters. PRESCILA prototype probes have been extensively tested at both Los Alamos and the German Bureau of Standards, Physikalisch-Technische Bundesanstalt. Test results are presented for energy response, directional dependence, linearity, sensitivity, and gamma rejection. Initial field tests have been conducted at Los Alamos and these results are also given. It is concluded that PRESCILA offers a viable, ergonomically superior, alternative to traditional rem meters that is effective for a wide range of neutron fields. The probe is capable of excellent sensitivity (40 counts per minute per microSv h-1 for 241AmBe) and extended energy response to beyond 20 MeV. Directional response is uniform (+/-15%) over a wide range of energies. Response linearity has been characterized to over 20 mSv h-1. Gamma rejection is effective in gamma fields up to 2 mSv h-1. The PRESCILA technology has been commercialized and is now offered under license by Ludlum Measurements, Inc.

Equipment Design↗

Direct quantitative digital autoradiography-thin-layer chromatography of 3 alpha,3 beta- and 5 alpha-reduced and 17 beta-dehydrogenated androgens derived from testosterone metabolism.

A digital autoradiographic-thin-layer chromatographic method involving simple steps is described for thorough separation of eight major androgens (testosterone, androstenedione, dihydrotestosterone, androsterone, epiandrosterone, androstanedione, 3 alpha-androstanediol and 3 beta-androstanediol) derived from testosterone metabolism. Their direct quantification is performed by radioscanning, which avoids tedious plate-scraping and liquid scintillation counting. Satisfactory accuracy is obtained by both external standardization of plates by calibrated amounts of radiolabelled standard steroids and internal standardization of bioassays by radiolabelled squalane. Coefficients of variation are below 8.5% in the range 100-500 dpm. Some analytical criteria related to chromatographic conditions and quantification parameters depending on position-sensitive proportional counter are discussed.

Androgens↗

Some applications of the indium-113m-transferrin technique for gamma-radiation detection of microvascular effects of serotonin and ischemia.

The distribution of systematically injected In-113m (t1/2 = 100 min) in organs of the rat was analyzed, and the use of the isotope for in vivo and in vitro gamma-radiation detection studies of blood plasma protein extravasation was demonstrated in skin, muscle, and tumor. In-113m was slowly excreted from rats. One to 6 h after injection the blood held 3% and 2%, respectively, of injected radioactivity/g tissue wet weight; skin and muscle held 0.1%-0.2%/g; liver, colon, and spleen held approximately 1%/g; lungs 1.5%-1.3%/g and kidneys 2.8%-3.3%/g. Scintillation camera technique revealed 40%-80% extraaccumulation of In-113m in a control extremity upon local administration of serotonin and 20%-40% in an extremity with a transplanted tumor, thus indicating a lower effect of serotonin in tumor microvascular circulation than in muscle and skin. In vitro detection of In-113m radiation by a well-counter in dissected tissues showed no effects of serotonin in the tumor and a four- to five-fold increase of radioactivity in muscle and skin, thus confirming blood protein extravasation upon serotonin treatment in these tissues. External analyses of In-113m in the vascular system using one miniaturized probe directed toward an area of interest showed that the method was too sensitive to movements of the animal, and second probe directed toward a control area is needed.

Animals↗

The fate of four 75Se-labeled amino acids: studies of some unsuccessful pancreas-scanning agents.

The Radiochemical Centre developed a number of alpha-amino acids labeled with 75Se in the hope of finding a pancreas-scanning agent superior to L-selenomethionine. Six were submitted for clinical trial and we tested four of these: D-selenomethionine and three derivatives of selenocysteine. Dynamic scintillation camera studies showed that liver uptake was substantial in all cases but that the time course of accumulation and removal differed between agents. Plasma radioactivity curves demonstrated differences in initial removal rates. Two of the agents showed evidence of incorporation of activity into protein and release into the circulation just as with L-selenomethionine. Whole-body counter measurements showed that the retention of activity from the agents exhibiting early plasma protein incorporation was similar to that of L-selenomethionine. Much of the activity from the other two agents was excreted faster but small proportion was retained for a long time. Although the cummulated activity and absorbed dose were substantially less for at least two of the agents than for 75Se-L-selenomethionine, the lack of sufficient pancreatic concentration renders them inferior as scanning agents.

Amino Acids↗