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PubMed2004

High radioactive concentration of 99mTc from a zirconium [99Mo]molybdate gel generator using an acidic alumina column for purification and concentration.

BACKGROUND: Newer applications of radiopharmaceuticals in nuclear medicine require pertechnetate of moderate to high radioactive concentration. Hence there is a need to develop simple procedures for the concentration of pertechnetate, and such a procedure is given in this paper. METHODS: Ten to 20 ml of sodium [Tc]pertechnetate eluted in de-ionized water from a zirconium [Mo]molybdate (ZrMo) gel column generator was passed through 2 g of an acidic alumina bed (35 x 8 mm) in order to remove the co-eluted traces of Mo and to retain the pertechnetate. The retained pertechnetate was then re-eluted, quantitatively, in 3 ml of normal saline, from the alumina column. RESULTS: About a 4-fold increase in radioactive concentration of Tc was obtained (cf. approximately 10-12 ml normal saline is required for the elution of Tc from the gel column). Generators containing up to 22.2 GBq (600 mCi) Mo in 6-7 g ZrMo gel column (35 x 13 mm) were prepared and a radioactive concentration of Tc up to 4 GBq x ml (110 mCi x ml) was obtained on the first day of use. The overall recovery of Tc was >90%, Mo breakthrough was 10 to 10% and the duration of concentration was 3-5 min. The chemical impurity in terms of Al, Mo and Zr was <10 ppm each. The same procedure for the concentration of pertechnetate was applied to generators with 12-15 g ZrMo gel beds to obtain a higher capacity Tc gel generator, with similar findings.

Aluminum Oxide↗
PubMed1975

Radioactive labelling of borme mosaic virus.

Brome mosaic virus (BMV) can be radioactively labelled by allowing virus synthesis to take place in barley leaves whose cut ends are placed in solutions of radio active precursors of BMV. When barley plants are infected by inoculation of the first emerging leaf, the rate of incorporation of both [14C]-leucine and [32P]-ortho-phosphate into virus particles was found to be maximal in the second leaf, cut and labelled 4 to 5 days after inoculation. The rate of incorporation declined when the labelling or the cutting and labelling were delayed, even though assembly of particles still took place from pre-formed components. An exposure of 72 h to an appropriate amount of 32P yielded highly radioactive BMV RNA for possible sequencing studies. A specific radioactivity (32P) of the order of 1 mCi/mg RNA can be obtained readily.

Adenosine↗
PubMed1989

Galactolipids of Thylakoid Pigment Protein Complexes Separated Electrophoretically from Thylakoids of Dunaliella salina Labeled with Radioactive Fatty Acids.

Experiments have been conducted to determine whether mono- and digalactosyldiacylglycerols (MGDG and DGDG) labeled by short term exposure of the green alga Dunaliella salina to exogenous radioactive lauric, myristic, or palmitic acids are preferentially recovered in one or more thylakoid protein complexes separated by electrophoresis on nondenaturing gels. A selective retention of certain lipid classes and molecular species by the major electrophoretically separable fractions, oligomeric form (LHCP-1) and monomeric form (LHCP-3) of hight-harvesting chlorophyll a/b protein complex, and major photosystem I protein complex (CPla), and nonprotein-bound lipids, was indicated. After 5 min of labeling with (3)H-palmitate the specific radioactivities of MGDG and DGDG molecular species recovered from LHCP-3 and CPla were slightly higher and lower, respectively, than specific radioactivities of equivalent molecular species from LHCP-1 and nonprotein-bound lipid. However, the differences were smaller than those which have been observed between whole thylakoids and the more highly labeled chloroplast envelopes from briefly labeled cells.

Journal Article↗
PubMed1992

Simple Determination of the CO(2)/O(2) Specificity of Ribulose-1,5-Bisphosphate Carboxylase/Oxygenase by the Specific Radioactivity of [C]Glycerate 3-Phosphate.

A new method is presented for measurement of the CO(2)/O(2) specificity factor of ribulose-1,5-bisphosphate carboxylase/oxygenase (Rubisco). The [(14)C]3-phosphoglycerate (PGA) from the Rubisco carboxylase reaction and its dilution by the Rubisco oxygenase reaction was monitored by directly measuring the specific radioactivity of PGA. (14)CO(2) fixation with Rubisco occurred under two reaction conditions: carboxylase with oxygenase with 40 micromolar CO(2) in O(2)-saturated water and carboxylase only with 160 micromolar CO(2) under N(2). Detection of the specific radioactivity used the amount of PGA as obtained from the peak area, which was determined by pulsed amperometry following separation by high-performance anion exchange chromatography and the radioactive counts of the [(14)C]PGA in the same peak. The specificity factor of Rubisco from spinach (Spinacia oleracea L.) (93 +/- 4), from the green alga Chlamydomonas reinhardtii (66 +/- 1), and from the photosynthetic bacterium Rhodospirillum rubrum (13) were comparable with the published values measured by different methods.

Journal Article↗
PubMed1998

Soft X-ray beamline specialized for actinides and radioactive materials equipped with a variably polarizing undulator.

This report presents the design of an undulator beamline at SPring-8 to be used for soft X-ray spectroscopy focused on radioactive materials. Photoemission spectroscopy experiments are carried out in a radioisotope (RI)-controlled area where actinide compounds as well as unsealed radioactive materials are usable. Intrusion of the radioactive materials into the electron storage ring or to the outside of the evacuated beamline components can be avoided by a specially devised RI protection/inspection mechanism. The combination of a variably polarizing undulator and a varied-line-spacing plane-grating monochromator provides linearly or circularly polarized soft X-rays with a high resolving power in the energy range 0.28-1.5 keV. The beamline will become operational in December 1997.

Journal Article↗
PubMed1984

Radioactive labels for Protein A: evaluation in the indirect immunoradiometric assay (IRMA) for Bacillus anthracis spores.

Staphylococcus aureus Protein A (SpA) labelled with [125I] by the Bolton & Hunter (1973) method performed about as well as labelled sheep anti-rabbit globulin (SAR) in an indirect immunoradiometric assay (IRMA) for Bacillus anthracis spores immobilized on multispot microscope slides. SpA labelled with [3H] by propionylation also performed well but would be expensive to use. SpA labelled with [3H] fluorodinitrobenzene, or labelled with [125I] by the chloramine T reaction gave erratic assay results, high noise values and low signal-to-noise ratios, indicating substantial direct binding of labelled SpA to the slide surface and to the bacterial preparation. The uptake of radioactively labelled SpA in the IRMA was compared with the fluorescence intensity of individual spores in a microfluorometric immunofluorescence (IF) test involving dual labelled fluorescein-[125I]-SpA. The maximum number of SAR molecules bound to the mixture of spores and cell-free antigens in the B. anthracis IRMA was about twice the maximum number of radioactively labelled SpA molecules bound. The SAR:SpA saturation binding ratio on the surface of the spores, however, was approximately the inverse of this. It is concluded that radioactively-labelled SpA is not recommended in preference to anti-species antibody reagents in bacterial IRMA tests but fluorescein-conjugated SpA deserves further consideration for use in microscope-based IF tests for bacterial antigens.

Antibodies, Anti-Idiotypic↗
PubMed1979

Yeast tRNA Leu UAG. Purification, properties and determination of the nucleotide sequence by radioactive derivative methods.

A second major species of leucine tRNA, tRNA Leu UAG (formerly designated tRNA Leu CUA) was purified from baker's yeast in a three-step procedure entailing BD-cellulose chromatography in the presence and absence of Mg2+ and Sephadex G-100 gel filtration. Results of aminoacylation and partial RNase T1 digestion experiments showed that this tRNA retains a native conformation under conditions that denature yeast tRNA Leu m5CAA (tRNA3 Leu). The primary structure of baker's yeast tRNA Leu UAG was elucidated by application of sensitive radioactive isotope derivative ("postlabeling") methods. Complete RNase T1 and A and partial RNase U2 fragments, prepared from non-radioactive tRNA and 5'-half and 3'-half molecules, were separated by two-dimensional polyethyleneimine-cellulose anion-exchange thin-layer chromatography and isolated by a novel micropreparative procedure affording high yields of these compounds in sufficient purity for subsequent tritium derivative analysis. Base composition and sequence of oligonucleotides were analyzed by tritium derivative methods. Molar ratios of the fragments were determined from the radioactivity of 3H-labeled nucleoside trialcohols in combination with base analysis. 2'-O-Methylated guanosine was characterized using the [gamma-32P]ATP/polynucleotide kinase reaction. The analysis of classical complete and partial RNase digests by the tritium derivative methods yielded the complete nucleotide sequence of the tRNA. A total of about 20 A260 units of the RNA was used for analysis, i.e. considerably less material than required for conventional spectrophotometric analysis. A different sequencing approach, consisting of a combination of "readout sequencing" with tritium sequencing of complete RNase T1 and A fragments, was applied to the 3'-half molecule. The 3'-half molecule was labeled with 32P at its 5' terminus, partially degraded with RNase T1, U2, and Phy1 and with alkali, and subjected to polyacrylamide gel electrophoresis. The sequence was read off the gel on the basis of cleavage patterns and size of the fragments. While the readout procedure provided only the positions of A, U, C, and G residues in the chain, additional information from tritium derivative analysis was utilized to define the positions of the modified nucleosides. The readout sequencing procedure was found to require less than 0.01 A260 unit of RNA and the analysis of the complete fragments about 6 A260 units. Interesting structural features of tRNA Leu UAG are (a) the location of unique, leucine tRNA iso-acceptor-specific sequences next to U-8, a constant nucleotide participating in synthetase recognition, (b) the occurrence of 1-methyladenosine in the T loop, a modification not present in the structurally related tRNA Leu m5CAA, and (c) the unusual presence of an unmodified uridine in the first position of the anticodon, which may be related to the unusual coding properties reported for this tRNA.

Adenine↗
PubMed1985

Autophagic-lysosomal and mitochondrial sequestration of [14C]sucrose. Density gradient distribution of sequestered radioactivity.

[14C]Sucrose, introduced into the cytosol of isolated rat hepatocytes by means of electropermeabilization, was sequestered by sedimentable subcellular particles during incubation of the cells at 37 degrees C. The sedimentation characteristics of particle-associated [14C]sucrose were different from the lysosomal marker enzyme acid phosphatase, suggesting an involvement of organelles of greater size than the average lysosome. Isopycnic banding in isotonic metrizamide/sucrose density gradients resolved two major peaks of radioactivity: a light peak (1.08-1.10 g/ml) coinciding with lysosomal marker enzymes, and a dense peak (1.15 g/ml), coinciding with a mitochondrial marker enzyme. The dense peak was preferentially associated with large-size particles having the sedimentation properties of mitochondria, and it was resistant to the detergent digitonin at a concentration which extracted all of the radioactivity in the light peak. Similarly the autophagy inhibitor 3-methyladenine prevented accumulation of [14C]sucrose in the light peak, while the radioactivity in the dense peak was unaffected. We therefore tentatively conclude that the light peak represents autophagic sequestration of [14C]sucrose into lysosomes (and probably autophagosomes) while the dense peak represents a mitochondrial uptake unrelated to autophagy.

Acid Phosphatase↗
PubMed1984

Radioactive choline uptake in the isolated rat phrenic nerve-hemidiaphragm preparation. A biochemical and autoradiographic study.

When hemidiaphragms are stimulated via the phrenic nerve in the presence of 10 microM radioactive choline (Ch), the rate of radioactive Ch uptake in the endplate-rich area (EPA) is greater than that in the endplate-poor muscle (M). Ch uptake in the EPA is temperature-dependent, with a Q10 of 2.9 and an activation energy of 19.5 kcal/mol. It is inhibited in a Na+-depleted medium, in the absence of Ca2+, and by 10-20 microM hemicholinium-3 (HC-3) and it is not inhibited by alpha-bungarotoxin even when the muscle is completely paralyzed. In the absence of stimulation the rate of uptake in the EPA is slightly, but not significantly, greater than in M. Using autoradiography, we find an enhanced amount of isotope in the nerve terminals and their immediate vicinities compared with the muscle fibres, in both stimulated and unstimulated hemidiaphragms. There is no enhanced uptake of isotope into the nerve terminals in stimulated tissues in the presence of 26 microM HC-3. The uptake of isotope into the muscle is not altered by any of these treatments. There is a positive correlation between the initial rate of radioactive Ch uptake in the EPA and the amount of isotope in the nerve terminals (the mean corrected grain density above the nerve terminals). Without correcting for the large amount of diffusion that occurs, the ratio of the grain density above the synapses to that above the muscle fibres is 1.66 in tissue stimulated at 1 Hz, 1.04 in stimulated tissues in the presence of 26 microM HC-3, and 1.31 in unstimulated tissues.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗
PubMed1987

Dynamic uptake of radioactive substance in rat salivary gland following 3H-melatonin administration.

Dynamics of radioactive accumulation in rat greater salivary gland following systemic administration of 3H-melatonin was studied to determine a possible action of the hormone in the gland. Progressive decline of 3H-melatonin concentrations was found in the serum, lung, skeletal muscle, liver, kidney, and salivary gland during 60 min following the administration. On the contrary, there was a progressive accumulation of radioactive substance other than 3H-melatonin in the salivary gland but not in other tissues mentioned. The radioactivity was also progressively and preferentially localized in the nuclear fraction of the gland cells. These results suggest a possible direct action of melatonin derivative in rat salivary gland.

Animals↗
PubMedDate not supplied

Identifying removable radioactivity on the surface of cats during the first week after treatment with iodine 131.

Because radioiodine (1-131) is excreted in urine and saliva, treated cats can accumulate I-131 on their coats from contacting soiled litter and grooming. This could result in removable radioactivity, which is a potential source of human exposure to radiation and specifically to internal contamination. The purpose of this study was to determine if there is removable radioactivity on cats treated with I-131. Daily wipe tests were performed for 7 days at two sites (both flanks, one site; and all four paws, one site) on six hyperthyroid cats treated with I-131. A y counter was used to determine the counts per minute (cpm) of the samples, which were converted to disintegrations per minute (dpm) to estimate activity. The results were compared to the New York State limits of removable activity for a non-controlled area (<1000dpm/100 cm2) to determine if the amount of removable activity was acceptable for a member of the public. The median value of removable activity was 241 dpm (range from 34 to 4184 dpm) for the flanks, and 308 dpm (range from 60 to 1890 dpm) for the paws. The amount of removable radioactivity on the surface of hospitalized cats treated with I-131 during the first week after treatment, occasionally and without obvious pattern, exceeded the New York State limit. Sporadic activity as high as 4148 dpm was found. It is prudent to advise owners to observe routine hygiene when handling cats after discharge to minimize the risk of internal contamination.

Animals↗
PubMed1979

Treatment of periocular tumours of horses using radioactive gold198 grains.

The literature is briefly reviewed concerning the clinical features of equine cutaneous tumours. The data from 19 horses with periocular skin tumours are reported together with the results of their treatment by implantation with radioactive gold198. The principles governing the implantation of radioactive sources are explained and a method for computing the initial activity of the sources is outlined. Five histologically different tumour types were treated in this series of 19 horses, 2 horses were bilaterally affected. The techniques of implantation using a simple needle/stillette combination and a purpose built gold grain implantation gun are described and the postoperative course of the animals is traced. The treatment was successful in ablating the tumour mass and preventing regrowth in 19 of 21 periocular tumour groups. The cosmetic effects of the treatment were found to be satisfactory. Emphasis is laid on the care needed in handling the radioactive material and on the need for a thorough understanding of the principles involved in radiotherapy. Doses received by the operator during an implantation procedure are quoted which show that the technique is safe as well as being effective.

Animals↗
PubMed1983

Enterohepatic circulation of radioactivity following an oral dose of [14C]temazepam in the rat.

The enterohepatic circulation of radioactive material after administering [14C]temazepam was evaluated in three sets of male Wistar strain rats connected in pairs by bile duct-duodenum cannulae. After a single oral dose (10 mg kg-1) to the donor rat, the excretion of radioactivity in the urine and faeces of both rats and in the bile of the recipient rat was determined. Mean total recovery of the administered radioactivity was 92.2%. Based on the amount remaining in the donor rat (gastrointestinal tract and faeces), 81.7% of the dose was absorbed by the donor. The total amount recovered from the recipient, 69.4% of original dose (85.1% of donor's absorbed dose), represented the amount excreted in the donor's bile. Similarly, 54.1% of the original dose (77.9% of the transferred biliary excretion from donor) was reabsorbed by the recipient, and the biliary excretion from this animal (45.9% original dose) accounted for 86.% of the amount reabsorbed.

Administration, Oral↗
PubMed1973

Pituitary gonadotrophs: nuclear concentration of radioactivity after injection of ( 3 H)testosterone.

Gonadotrophs and castration cells in the male rat pituitary showed nuclear concentration of radioactivity 1 hour after [1,2,6,7-(3)H]testosterone injection. Thyrotrophs and acidophils did not retain radioactivity; also the cells of the intermediate and posterior lobes did not accumulate radioactivity. The autoradiographic results suggest a direct and selective action of androgen on gonadotrophs, which contrasts with the action of estradiol which was shown earlier to bind not only to basophils but to acidophils and chromophobes as well.

Animals↗
PubMed1963

ELECTROPHORETIC STUDY OF THE INTERACTION OF RADIOACTIVE POLIOVIRUS WITH COMPONENTS OF CULTURED CELLS.

Thorne, H. V. (University of Minnesota, Minneapolis). Electrophoretic study of the interaction of radioactive poliovirus with components of cultured cells. J. Bacteriol. 85:1247-1255. 1963.-The interaction of P(32)-labeled poliovirus with the postnuclear fraction of HeLa cell homogenates was first examined using radioactivity to quantitate the reaction. The effects of virus and debris concentration, suspending medium, temperature, and time on the reaction were determined. Binding was independent of salt concentration up to 1.0 m and unaffected by calcium and magnesium ions at 10(-3)m. The postnuclear particulate components of homogenates of mammalian cells in culture were examined by sucrose gradient zone electrophoresis at pH 7.2 with a simple apparatus which permitted several simultaneous analyses. The distributions of components for different cell types were distinct and appeared to be influenced by conditions of culture. Addition of radioactive poliovirus to the homogenate before analysis was used to identify components with virus-binding activity. Activity of HeLa and human esophageal epithelium cell homogenates was found mainly in membranous fractions of relatively low electrophoretic mobility. Components with a broader spread of mobility were moderately active in cultured human amnion, but uncultured amnion had almost undetectable virus-binding activity and a distinct distribution of components. Rat heart cells and L cells did not bind poliovirus, but binding components were present in both ERK-1 and CRE rabbit lines.

Animals↗
PubMed1980

Radioactivity studies on 2 synovial specimens after radiation synovectomy with yttrium-90 silicate.

Synovial membranes surgically removed from the knees of 2 patients who had received radioactive yttrium-90 silicate have been examined. Autoradiographs showed that in both cases the activity was unevenly distributed over the synovium. One of the specimens, removed 7 weeks after administration of the isotope, was found to contain a slight degree of radioactivity, attributable in part to a long-lived radioactive contaminant.

Arthritis, Rheumatoid↗
PubMed2004

Analytical and diagnostic accuracy of "second generation" assays for thyrotrophin receptor antibodies with radioactive and chemiluminescent tracers.

AIMS: To investigate the analytical and diagnostic accuracy of thyrotrophin (TSH) receptor antibody assays using recombinant human TSH receptors. METHODS: Sera from 68 patients with Graves' disease, 23 patients with autoimmune thyroiditis, and 119 healthy controls were evaluated in four different laboratories using both radioactive and chemiluminescent tracers. Functional sensitivity, interlaboratory precision, optimal cutoff values for Graves' disease, and the correlation between the two methods were evaluated. RESULTS: Functional sensitivity was 0.98 IU/litre for both assays. Interlaboratory precision, expressed as per cent coefficient of variation over a wide range of antibody concentrations, varied from 5.7% to 15.1% for the radioligand, and from 6.6% to 19.9% for the chemiluminescence assay. The two methods (radioactive and chemiluminescent) were closely correlated. All the sera from untreated or relapsing patients with Graves' disease gave TSH receptor antibody values above 2.1 IU/litre, whereas in none of the healthy controls did values exceed 2.5 IU/litre. Receiver operating curve analysis allowed an optimal cutoff point to be defined at 1.99 IU/litre, according to a sensitivity of 100% and specificity of 99.1%. CONCLUSIONS: These data show the high analytical and diagnostic accuracy of the human TSH receptor assays, both with radioactive and chemiluminescent tracers, when both functional sensitivity and interlaboratory reproducibility are considered. These two methods could be proposed as first line diagnostic markers for Graves' disease.

Adult↗
PubMed1986

A comparison of the acylation of 1-acyl-sn-glycero-3-phosphoinositol and 1-acyl-sn-glycero-3-phosphocholine in neuronal nuclei in vitro using radioactive arachidonate and oleate.

A neuronal nuclear fraction (N1) was isolated from cerebral cortices of 15-day-old rabbits. Samples of N1 were incubated with a radioactive fatty acid ([3H]arachidonate or [14C]oleate), acylation cofactors, and 1-acyl-sn-glycero-3-phosphoinositol (1-acyl-GPI) or 1-acyl-sn-glycero-3-phosphocholine (1-acyl-GPC). In competition studies, both radioactive fatty acids were incubated with one lysophospholipid or the two lysophospholipids were incubated with one radioactive fatty acid. Using [3H] arachidonate and one lysophosphoglyceride, a maximal rate of incorporation into phosphatidylinositol (PI) was found at a relatively low concentration of 1-acyl-GPI (10 microM), while increasing rates of incorporation into phosphatidylcholine (PC) were seen with increasing concentrations of 1-acyl-GPC (to 65 microM). At low concentrations of lysophosphoglyceride (less than or equal to 25 microM) the rate of arachidonate incorporation into PI greatly exceeded rates of arachidonate incorporation into PC. This higher rate of arachidonate incorporation into PI was also seen in incubations where both lysophospholipids were present. For oleate, greater rates of incorporation into PC were found in comparison with rates of labelling of PI in assays using relatively high concentrations of one or both lysophospholipids. When comparing arachidonate and oleate, in assays with one or both fatty acids, the polyunsaturate showed at least threefold higher rates of incorporation into PI. For PC labelling higher rates of arachidonate incorporation were evident at the higher concentrations of 1-acyl-GPC and the superiority over oleate was not as marked as that seen in PI labelling.

Acylation↗