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PubMed1994

Diagnostic pharmaco-scintigraphy with hepatic intra-arterial technetium-99m macroaggregated albumin in the determination of tumour to non-tumour uptake ratio in hepatocellular carcinoma.

Between October 1990 and March 1993, 124 patients who had hepatocellular carcinoma (HCC) underwent diagnostic pharmaco-scintigraphy with hepatic intraarterial technetium-99m macroaggregated albumin (TcMAA) to determine the tumourous to non-tumourous liver tissue uptake ratio (T/N ratio). There were 110 males and 14 females. Ages ranged from 16 to 73 with a median of 55 years. The range of T/N ratio was 0.7 to 19.3 with a median of 3.8. 12 patients with inoperable HCC were subsequently selected by predetermined criteria to undergo treatment with hepatic intraarterial yttrium-90 microspheres and the T/N ratios in these patients were validated by beta probe dosimetry and liquid scintillation count of multiple liver biopsies. The T/N ratio determined by preoperative diagnostic TcMAA scan correlated well with intraoperative beta probe dosimetry, with coefficient of correlation r = 0.82. Preoperative TcMAA scan also correlated well with liquid scintillation count of biopsy specimens, with r = 0.96. We conclude that TcMAA scan can be used to determine the T/N ratio in patients with HCC, thus allowing better selection of patients with inoperable tumours for loco-regional therapy.

Adolescent↗
PubMed1992

Differential proliferation of rat aortic and mesenteric smooth muscle cells in culture.

Smooth muscle cells (SMC) from various arterial origins have been successfully maintained in culture. The present study evaluates the proliferative activity of aortic and mesenteric SMC in culture. Aortic and mesenteric SMC were obtained from male Wistar rats by explant and enzyme digestion techniques, respectively. Vascular SMC obtained by either method exhibited a characteristic hill-and-valley growth pattern in culture after confluence and were positively labelled with either anti-smooth muscle actin or myosin by an indirect immunofluorescent method. The rate of incorporation of thymidine into DNA and cell number counting were used as indices of proliferation in vitro. Vascular SMC from passages 4-33 were first synchronized with either Dullbecco's Modified Eagle's Medium (DME) or Ham's F-12 medium, supplemented with insulin-transferring-selenium (ITS), for 72 hours. SMC were then stimulated with 10% bovine serum for either 24 or 72 hours with the former processed for scintillation counting, the latter for cell number determination. The incorporation of tritiated thymidine into DNA following a 2 hour incubation was determined by scintillation counting after perchloric acid extraction. In terms of cell numbers, proliferative responses to bovine serum were determined by Coulter counting. Autoradiography was also carried out in some cultures to determine both thymidine and mitotic labelling indices. The rate of thymidine incorporation in aortic cells was 2-3 fold higher than in mesenteric cells. Aortic and mesenteric SMC lines exhibited similar cell cycle intervals in terms of total duration and individuals cycle parameters. However, the total thymidine index was higher in the aortic than mesenteric SMC.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗
PubMed2006

Standardization of radionuclide by beta(LS)-gamma coincidence counting using the geometry-efficiency variation method.

A liquid scintillation counting method consisting of three photomultiplier tubes for beta counters and a NaI(Tl) gamma counter has been developed for the standardization of radionuclides with the beta-gamma coincidence technique. The beta detection efficiency functions are obtained by means of a geometry-variation method developed in the present work; an array of beta detectors is moved uniformly at the same time from a centrally located counting vial to 50 mm. The method has been applied in the standardization of 60Co and 134Cs. Unquenched liquid scintillation samples with nominal count rates from 1000 to 6000 s-1 were prepared. The observed beta detection efficiencies with this method are from 90 to 45% in the case of 60Co, and from 84 to 50% for 134Cs. The output of each beta channel is summed together and compared with gamma data by the coincidence analyzer. The dead time of each counting channel is adjusted to be 20 micros, sufficiently long to suppress the afterpulses in the beta counting channel. The activity of each sample is determined by using the Cox and Isham formula. The obtained results are in good agreement with KRISS certified values.

Beta Particles↗
PubMed1992

Technique-dependent variations in cerebral microvessel blood volumes and hematocrits in the rat.

To quantitate small parenchymal microvessel blood volumes in the brain, the distribution spaces of radiolabeled red blood cells (RBC) and serum albumin (RISA) were assessed in rats by different methods of tissue sampling and radioassay. Three minutes after intravenous administration of 55Fe-RBCs and/or 125I-RISA, the rats were decapitated. The brain was either immediately frozen within the skull and later removed (head-frozen group) or rapidly removed from the skull and then frozen (brain-frozen group). Radioactivity was measured either by liquid scintillation counting of tissue pieces, which contained pial plus large and small parenchymal microvessels, or by quantitative autoradiography (QAR) of tissue sections, which indicated small parenchymal microvessel blood only. In 12 of 15 areas, the RISA, RBC, and blood volumes determined by liquid scintillation counting of head-frozen tissue pieces were equal to or greater than those of brain-frozen tissue; this indicated less than or equal to 25% greater blood retention in pial and parenchymal microvessels with head freezing. At the parenchymal microvessel level (QAR assay), the distribution volumes of RBCs, RISA, and blood were similar with the two freezing techniques; hence with QAR either freezing procedure can be used to assess small parenchymal microvessel blood volumes.

Animals↗
PubMed1996

Biodistribution and clearance of 99mTc-labeled Arg-Gly-Asp (RGD) peptide in rats with ischemic acute renal failure.

Based on the previous demonstration of a renoprotective effect of arginine-glycine-aspartic acid (RGD) peptides in acute renal failure, experiments were designed to test the distribution and renal accumulation of the peptide. To accomplish this goal, in this study, RGD peptide was radiolabeled and its biodistribution and renal accumulation was determined in rats with ischemic acute renal failure (ARF). 99mTc-RGD with or without 111In-DTPA were injected intravenously in control and ARF rats. Various organs were dissected at different times after injection and subjected to gamma-scintillation counting and autoradiography (ARG). Blood clearance of 99mTc-RGD was rapid, with t1/2 < 10 min, and unchanged in ARF compared with control rats. Kidneys retained the largest portion of the injected dose in both control and ARF rats, as detected using scintillation counting and whole-body ARG (10.56 +/- 1.05% and 10.12 +/- 3.16% injected dose/g wet weight, respectively). Renal ARG revealed a significant increase in binding to the cortex in ARF kidneys, compared with that of control kidneys. Given the differences in renal blood flow and GFR in control and postischemic kidneys, the next series of experiments was performed with two radiopharmaceuticals, 99mTc-RGD and 111In-DTPA. The ratio of 99mTc-RGD:111In-DTPA was increased more than three-fold in ARF kidneys compared with control kidneys (2.7 +/- 0.15 versus 0.8 +/- 0.19, respectively). The results indicate that (1) RGD peptide undergoes a rapid clearance predominantly via the renal route; (2) despite a significant reduction in the renal perfusion, 99mTc-RGD peptide accumulates in the postischemic kidney; (3) this is consistent with the hypothesis on the involvement of RGD-recognizing integrins in the development of tubular obstruction in renal ischemia.

Acute Kidney Injury↗
PubMed1976

The study of (Methyl-3H)decamethonium dichloride incorporation into normal and dystrophic mouse muscle.

1. (Methyl-3H)decamethonium dichloride was injected intravenously into the tail vein of dystrophic and normal mice of the Bar Harbour strain 129 ReJ/dy in paralysing doses. 2. Scintillation counts were made of 1 mm sections of diaphragm and tibialis anterior which showed a normal distribution of tritiated decamethonium in the dystrophic animals. 3. Intraperitoneal injections of L-(gamma-3H)leucine were made into dystrophic and normal mice. Examination of diaphragm and tibialis anterior by scintillation counting showed the abnormal uptake typical of dystrophic involvement of murine muscle. 4. The normal distribution of (methyl-3H)decamethonium dichloride uptake by the dystrophic muscle does not support the concept of active denervation in this disease.

Animals↗
PubMed1972

Technique for measuring 14 CO 2 uptake by soil microorganisms in situ.

Uptake of (14)CO(2) in soils due to algae or sulfur-oxidizing bacteria was examined by incubation of soil samples with gaseous (14)CO(2) and subsequent chemical oxidation of biologically fixed radioactive isotope to (14)CO(2) for detection with a liquid scintillation counting system. The (14)CO(2) was added to the soil in the gas phase so that no alteration of the moisture or ionic strength of the soil occurred. Wet oxidation of radioactive organic matter was carried out in sealed ampoules, and the (14)CO(2) produced was transferred to a phenethylamine-liquid scintillation counting system with a simply constructed apparatus. The technique is inexpensive and efficient and does not require elaborate traps since several possible interfering factors were found to have no harmful effects. Experiments in coal mine regions and in geothermal habitats have demonstrated the ecological applicability of this technique for measurement of CO(2) fixation by sulfur-oxidizing bacteria and soil algae.

Bacteria↗
PubMed1987

Quantitation of genes and transcripts by saturation hybridization in urea solutions using strand-selected probes.

This report describes methods for quantifying specific sequences in preparations of single-stranded or double-stranded nucleic acids. We use saturating amounts of hybrid-selected, strand-specific probes and perform hybridizations in urea solutions. Hybrids (RNA-DNA or DNA-DNA) are then analyzed by either of two methods. The first employs standard S1 nuclease digestion, precipitation of resistant material on glass fiber filters, and assay by liquid scintillation counting. This method is generally chosen in the assay of rare transcripts in total nucleic acid extracts as well as preparations of polyadenylated RNA. The second employs the separation of excess probe from probe-target hybrids by gel electrophoresis, recovery of hybrids on ion-exchange paper, and determination of cpm bound by liquid scintillation counting. This method is of particular value in the assay of double-stranded sequences and the quantitation of restriction fragment length polymorphisms. Whereas both methods are accurate over ranges of target abundance representing at least several orders of magnitude, the latter ("NA45 assay") is most sensitive, and the selection of M13-bound or unbound DNA fragments by this method can be exploited in a variety of other applications.

Animals↗
PubMed1983

Extension of the ELISA method to the measurement of the specific radioactivity of viruses in crude cellular extracts.

The double-antibody sandwich method of ELISA, which allows accurate quantitative determination of plant viruses, was extended to a radiochemical procedure which permits direct measurement of the specific radioactivity of virus labelled in vivo and present in very crude plant homogenates. Evidence is presented showing that 20 to 50% of the virus introduced in the polystyrene wells during the antigen incubation step could be trapped in the sandwich. The percentage of virus bound increased with the concentration of the coating antibody and was almost proportional to the concentration of the antigen and to the incubation time of the antigens. Complete dissociation of the double-antibody sandwich was achieved by incubation with 0.2 M KOH or NaOH (pH 13.3), and the label carried by the virus was measured by scintillation counting of the solubilization fluid. The ratio infected/healthy was much higher for the radiochemical procedure than for the immunosorbent assay itself since binding of the virus to the coating antibody was not accompanied by any nonspecific trapping of radioactive contaminants in the double-antibody sandwich. The procedure was highly sensitive since the background corresponded to the scintillation counting background. The detection of label carried by tobacco mosaic virus was possible when the tobacco samples contained at least 5 ng of virus carrying a label as low as 40 dpm 3H or 20 dpm 14C.

Enzyme-Linked Immunosorbent Assay↗
PubMed1990

Tissue distribution of doxorubicin associated with polyisohexylcyanoacrylate nanoparticles.

The body distribution of i.v. doxorubicin depends mainly on the physicochemical characteristics of the molecule. However, entrapment of that cytostatic drug inside polyalkylcyanoacrylate nanoparticles has been shown to modify its distribution profoundly in the mouse. Polysiohexylcyanoacrylate nanoparticles loaded with [14C]-doxorubicin were studied in comparison with free drug, with emphasis on their distribution pattern in mouse tissue after i.v. administration. An autoradiographic study showed that most of the radioactivity was found in the reticuloendothelial system as soon as a few minutes after i.v. administration of the doxorubicin-loaded nanoparticles. Quantitative determinations by liquid scintillation counting in fresh tissue (spleen, heart, kidneys, liver, lungs, bone marrow) and blood samples confirmed these observations. When the drug was linked to nanoparticles, doxorubicin blood clearance was reduced during the first few minutes after administration, whereas heart and kidney concentrations were substantially decreased. Assays of doxorubicin and doxorubicinol by a specific HPLC analytical method gave results very similar to those obtained by scintillation counting.

Animals↗
PubMed2003

Controlled high meat diets do not affect calcium retention or indices of bone status in healthy postmenopausal women.

Calcium balance is decreased by an increased intake of purified proteins, although the effects of common dietary sources of protein (like meat) on calcium economy remain controversial. We compared the effects of several weeks of controlled high and low meat diets on body calcium retention, using sensitive radiotracer and whole body scintillation counting methodology. Healthy postmenopausal women (n = 15) consumed diets with similar calcium content (approximately 600 mg), but either low or high in meat (12 vs. 20% of energy as protein) for 8 wk each, in a randomized crossover design. After 4 wk of equilibration of each diet, calcium retention was measured by extrinsically labeling the 2-d menu with (47)Ca, followed by whole body scintillation counting for 28 d. Urinary and blood indicators of bone metabolism were also determined for each diet. Calcium retention was not different during the high and low meat dietary periods (d 28, mean +/- pooled SD: 17.1 and 15.6%, +/-0.6%, respectively; P = 0.09). An initially higher renal acid excretion in subjects consuming the high meat compared with the low meat diet decreased significantly with time. The diets did not affect urinary calcium loss or indicators of bone metabolism. In conclusion, under controlled conditions, a high meat compared with a low meat diet for 8 wk did not affect calcium retention or biomarkers of bone metabolism in healthy postmenopausal women. Calcium retention is not reduced when subjects consume a high protein diet from common dietary sources such as meat.

Aged↗
PubMed1972

Quantitative features of a sandwich radioimmunolabeling technique for lymphocyte surface receptors.

The present study was designed to devise and characterize an indirect or sandwich radioimmunolabeling technique for the study of lymphocyte surface receptors of immunoglobulin nature. Mouse lymphocytes from various sources were treated by the method of Shortman et al. to remove debris and damaged cells. This was an important preliminary step, as without it, little meaning could be attached to bulk scintillation counting of labeled cell suspensions, in view of the marked tendency of dead or damaged cells to adsorb protein nonspecifically. Next, cells were reacted at 0 degrees C for 30 min with graded dilutions of unlabeled rabbit antisera against defined mouse Ig chains. After washing, the cells were reacted with a sheep anti-rabbit globulin reagent labeled with (125)I, again at graded concentrations. After further washing, lymphocyte labeling was quantitated by both bulk scintillation counting and radioautography. Conditions were defined in which nonthymus-derived cells (B cells) but not thymus-derived cells (T cells) could be labeled. Most B cells displayed kappa- and micro-chains on their surface, but some also displayed alpha- and gamma(2)-chains, though in smaller amounts. When the concentration of both the first and the second reagents were raised considerably, conditions were defined under which virtually all T cells could be labeled by polyvalent antiglobulin sera, anti-kappa sera, or, with more difficulty, by anti-micro sera. A large series of control experiments confirmed the serologic specificity of this labeling. It was shown that under equivalent conditions, B cells bind 100-400 times more antiglobulin than do T cells. The theoretical implications of the results are briefly discussed. It is argued that the sandwich approach offers certain technical advantages over direct labeling procedures for further analyses of T cell receptors and for studies of receptor metabolism.

Animals↗
PubMedDate not supplied

Determination of malachite green residues in the eggs, fry, and adult muscle tissue of rainbow trout (Oncorhynchus mykiss).

Malachite green, an effective antifungal therapeutant used in fish culture, is a known teratogen. We developed a method to simultaneously detect both the chromatic and leuco forms of malachite green residues in the eggs, fry, and adult muscle tissue of rainbow trout (Oncorhynchus mykiss). Homogenates of these tissues were fortified with [14C] malachite green chloride and extracted with 1% (v/v) acetic acid in acetonitrile or in methanol. The extracts were partitioned with chloroform, dried, redissolved in mobile phase, and analyzed by liquid chromatography (LC) with postcolumn oxidation of leuco malachite green to the chromatic form. LC fractions were collected every 30 s for quantitation by scintillation counting. Recoveries of total [14C] malachite green chloride residue were 85 and 98% in eggs fortified with labeled malachite green at concentrations of 0.5 and 1.00 microgram/g, respectively; 68% in fry similarly fortified at a concentration of 0.65 microgram/g; and 66% in muscle homogenate similarly fortified at a level of 1.00 microgram/g. The method was tested under operational conditions by exposing adult rainbow trout to 1.00 mg/L [14C] malachite green chloride bath for 1 h. Muscle samples analyzed by sample oxidation and scintillation counting contained 1.3 and 0.5 microgram/g total malachite green chloride residues immediately after exposure and after a 5-day withdrawal period, respectively.

Animals↗
PubMed1997

Mangafodipir trisodium injection, a new contrast medium for magnetic resonance imaging: in vitro metabolism and protein binding studies of the active component MnDPDP in human blood.

The binding to human serum proteins of MnDPDP (manganese(II) dipyridoxyl diphosphate), the active component of the magnetic resonance imaging contrast medium mangafodipir trisodium injection (Teslascan) was studied in ultrafiltration experiments. Sera from three males and three females were incubated with 86 microM [14C]MnDPDP for 60 min at room temperature (20-23 degrees C), followed by centrifugation through filters with a cut-off of 30 kDa. Analysis of the filtrates and the initial incubation mixtures for manganese, by ICP-AES, and for DPDP and its dephosphorylated metabolites DPMP (dipyridoxyl monophosphate) and PLED (dipyridoxyl ethylenediamine diacetate) by liquid scintillation counting, showed a clear difference in protein binding of manganese and the ligands under these conditions. Only 2.2 +/- 1.8% (mean +/- S.E.; n = 6) of DPDP, DPMP and PLED were bound to protein, whereas 26.9 +/- 2.9% (mean +/- S.E.; n = 6) of manganese was bound to protein. No binding of DPDP, DPMP or PLED to blood cells was observed when whole blood, containing either heparin or EDTA as anticoagulant, was spiked with [14C]MnDPDP and the cell-free fraction and the lysed cell fraction analysed by liquid scintillation counting. The extent of protein binding of manganese corresponded well with results from an in vitro metabolism study, in which MnDPDP was added to heparinized human whole blood, showing that approximately 25% of DPDP, DPMP or PLED were not bound to manganese. The in vitro metabolism study revealed that transmetallation with zinc was nearly complete within 1 min, and that dephosphorylation is a sequential process going from DPDP to the monophosphate DPMP, and then to the fully dephosphorylated compound PLED.

Blood Proteins↗
PubMed1994

Polyacrylamide gradient gel electrophoresis as a method to measure transfers of radiolabeled cholesteryl esters between several plasma lipoprotein fractions.

In human plasma, cholesteryl esters can redistribute between various lipoprotein fractions through an exchange reaction mediated by the cholesteryl ester transfer protein. In the present study, liquid scintillation counting of polyacrylamide gradient gel fragments was applied to the determination of transfers of radio-labeled cholesteryl esters between different lipoprotein fractions. This method consisted in four successive steps: (1) incubation of total human plasmas with a tracer dose of high-density lipoproteins (HDL) containing tritium 3H-labeled cholesteryl esters, (2) separation of plasma lipoprotein fractions by electrophoresis in 20-160 g/liter polyacrylamide gels, (3) cutting off and dissolution with a NaOCl solution of polyacrylamide gel fragments which contained the different lipoprotein fractions, (4) liquid scintillation counting of the dissolved gel fragments. The present method provided a convenient and accurate method of determining the rate of cholesteryl ester transferred between several plasma lipoprotein classes. As an example, it was applied to the measurement of the rates of radiolabeled cholesteryl esters transferred from either HDL3 or HDL2 toward the two other main plasma lipoprotein fractions, very-low-density lipoproteins and low-density lipoproteins.

Cholesterol Esters↗
PubMed1997

A proposed method to minimize waste from institutional radiation safety surveillance programs through the application of expected value statistics.

Institutional radiation safety programs routinely use wipe test sampling and liquid scintillation counting analysis to indicate the presence of removable radioactive contamination. Significant volumes of liquid waste can be generated by such surveillance activities, and the subsequent disposal of these materials can sometimes be difficult and costly. In settings where large numbers of negative results are regularly obtained, the limited grouping of samples for analysis based on expected value statistical techniques is possible. To demonstrate the plausibility of the approach, single wipe samples exposed to varying amounts of contamination were analyzed concurrently with nine non-contaminated samples. Although the sample grouping inevitably leads to increased quenching with liquid scintillation counting systems, the effect did not impact the ability to detect removable contamination in amounts well below recommended action levels. Opportunities to further improve this cost effective semi-quantitative screening procedure are described, including improvements in sample collection procedures, enhancing sample-counting media contact through mixing and extending elution periods, increasing sample counting times, and adjusting institutional action levels.

Radiation Monitoring↗
PubMedDate not supplied

The measurement of technetium-99 and iodine-129 in waste water from pressurized nuclear-power reactors.

The program for measuring the dispersal presence of contaminating nuclides in waste streams produced by pressurized-water-reactor (PWR) power plants required the establishment of analytical procedures for a number of nuclides. In this report the procedure to assess 99Tc and 129I content in liquid samples using liquid-scintillation counting is presented. 99Tc and 129I are adsorbed on AG 1 X2 ion-exchange resin. 129I is then eluted with NaOCl and further purified by liquid-liquid extraction using carbon tetrachloride and an aqueous solution of sodium thiosulphate. 99Tc is eluted with hot nitric acid (6 M/60 degrees C). If necessary any 60Co present in the sample is separated by liquid-liquid extraction of the cobalt-TTA complex with iso-BuOH. The overall recovery yield is 96.5 +/- 2.5% for 99Tc and 90.1 +/- 6.2% for 129I. Finally beta-activity is measured by liquid-scintillation counting. A limit of detection of 5 Bq/L can thus be obtained for both 99Tc and 129I.

Belgium↗
PubMed2002

Convection-enhanced delivery of boronated epidermal growth factor for molecular targeting of EGF receptor-positive gliomas.

Convection enhanced delivery (CED) is potentially a powerful method to improvethe targeting of macromolecules to the central nervous system by applying a pressure gradient to establish bulk flow through the brain interstitium during infusion. The purpose of the present study was to evaluate CED as a means to improve the intracerebral and intratumoral (i.t.) uptake of a heavily boronated macromolecule (dendrimer; BD) linked to epidermal growth factor (EGF) for neutron capture therapy in rats bearing a syngeneic epidermal growth factor receptor (EGFR) + glioma. Boronated EGF was radiolabeled with 125I and administered by CED at a rate of 0.33 micro l/min for 15, 30, and 60 min [infusion volumes (V(I)) of 5, 10, and 20 micro l, respectively], using a syringe pump connected to an indwelling cannula implanted into the right caudate nucleus of normal rats or i.t. in rats bearing either F98(EGFR) or F98 wild-type (F98(WT)) gliomas. After infusion, rats were euthanized, and their brains were removed and serially sectioned. The uptake and biodistribution of (125)I-boronated EGF in tumor or brain was studied by quantitative autoradiography and gamma-scintillation counting. The volume of distribution (V(d)) in brain was assessed using a computer interfaced image analysis system. After CED, the V(d) increased from 34.4 to 123.5 micro l with corresponding V(i) ranging from 5 to 20 micro l. The V(d) of BD-EGF in the brain was 64.8 +/- 13.4 micro l with CED (V(i) 10 micro ), and the V(d):V(i) ratio was 6.5 compared with a V(d) of 9.4 +/- 1.6 micro l and a V(d):V(i) ratio of 0.9 after direct intracerebral injection. As determined by quantitative autoradiography and gamma-scintillation counting at 24 h after CED, 47.4% of the injected dose per gram tissue (%ID/g) was localized in F98(EGFR) gliomas compared with 33.2%ID/g after direct i.t. injection and 12.3%ID/g in F98(WT) gliomas. On the basis of these observations, we have concluded that CED is more effective than i.t. injection as a way to deliver boronated EGF to EGFR (+) gliomas for boron neutron capture therapy.

Animals↗