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Disposition of radioactivity after injection of liver-targeted proteins labeled with 111In or 125I. Effect of labeling on distribution and excretion of radioactivity in rats.

The effect of radiolabeling liver-specific proteins on the in vivo disposition of radioactivity was investigated. The suitability of 111In and 125I as radiolabels for protein disposition studies in vivo was examined. Galactosylated and cationized bovine serum albumin were labeled with either 125I by the chloramine-T method or 111In, using 1-(4-isothiocyanatobenzyl)ethylenediaminetetraacetic acid (SCN-BZ-EDTA) or diethylenetriaminepentaacetic acid (DTPA) as bifunctional chelating agents (BCAs) and administered intravenously to rats. 125I radioactivity disappeared rapidly from the liver with subsequent excretion in the urine and bile, mainly in the TCA soluble fraction. 111In-associated radioactivity, on the other hand, remained in the hepatic tissue in considerably higher amounts during the experiment and was excreted in the bile and urine to a lower extent when compared with 125I. When the effect of BCA on excretion of 111In radioactivity was compared, no significant differences were observed in the urinary clearances. However, biliary excretion was significantly higher for 111In-SCN-BZ-EDTA-bound radioactivity. In conclusion, when compared with 125I, 111In labeling seems to more accurately characterize the in vivo distribution of liver-targeted proteins after their iv administration in rats and allows a more accurate pharmacokinetic evaluation to be performed.

Albumins↗

The incorporation of radioactive fatty acids and of radioactive derivatives of glucose into the phospholipids of subsynaptosomal fractions of cerebral cortex.

1. Crude synaptosomal fractions (P2) from guinea-pig cerebral cortex were incubated in a Krebs-glucose medium containing labelled fatty acids and [3H]glucose. After the shortest incubation period (7.5 min) a high percentage (50-80%) of the total radioactive fatty acids was found in the P2 fractions. 2. After the incubation, the synaptosomal fractions were submitted to hypo-osmotic disruption and subsynaptosomal fractionation was carried out by using discontinuous-sucrose-gradient centrifugation. The specific radioactivities of phosphatidylcholine, phosphatidylethanolamine, phosphatidylserine and phosphatidylinositol were determined in fractions D (synaptic vesicles), E (microsomal preparation) and H (disrupted synaptosomes), as were the specific activities of a number of marker enzymes and the distribution of acetylcholine. 3. By using [14C]oleate, [14C]arachidonate, [3H]palmitate and [3H]glucose, the order to specific radioactivities in fraction D was found to be: phosphatidylinositol greater than phosphatidylcholine greater than phosphatidylserine greater than phosphatidylethanolamine. 4. The specific radioactivities of phosphatidylcholine and phosphatidylethanolamine were always higher in fraction D than in fraction E. As fraction E had higher specific activities of several membrane marker enzymes, the enhanced labelling found in fraction D was considered to be localized in the synaptic vesicles. In this fraction, phosphatidylinositol made particularly large contributions to the total phospholipid labelling derived from [14C]arachidonate and [3H]glucose. 5. The similar labelling ratios of fatty acid/glucose in the phospholipids of fractions D and E, and the high specific radioactivities in the total phospholipid of the soluble fraction O, suggested intrasynaptosomal phospholipid transport.

Acetylcholine↗

Transfer of radioactive materials from radioactively labeled conditioned medium to fixed cells.

Transfer of radioactive materials to glutaraldehyde-fixed Ni12Cl cells was observed when fixed cells were incubated in conditioned media of Ni12Cl cells cultured with various radioactive precursors such as glucosamine, fucose or leucine. Radioactivities were not transferred to fixed cells when the conditioned medium was boiled. The transfer decreased drastically by treatments of the medium with trypsin or detergents (dodecylsulfate or deoxycholate). Efficient transfer occurred only at the physiological temperature. No radioactivity was transferred to fixed cells if conditioned media of nonlabeled cells and radioactive precursors including various nucleotide sugars were incubated with fixed cells. The transferring activity in a conditioned medium did not decrease by the dialysis or the centrifugation at 105,000 x g for 2 h, and remained in the supernatant. The activity migrated into the sucrose gradient. Materials transferred to fixed cells were not solubilized by detergents but were released by the treatment with trypsin, indicating transferred materials were covalently associated with fixed cells. Those results suggest that conditioned media thus obtained contain both substrate(s) and enzyme(s) which fixed labels to fixed cells.

Animals↗

Tissue distribution of radioactivity following intranasal administration of radioactive microspheres.

The aim of this study was to increase understanding of the kinetics of microparticle distribution and elimination following intranasal application. To do this we investigated the in-vivo distribution of radioactivity following intranasal instillation of scandium-46 labelled styrene-divinyl benzene 7-microm-diameter microspheres. Groups of BALB/c mice received 0.250 mg (47.5 kBq) particles suspended in either 50-microL or 10-microL volumes of phosphate buffered saline. The in-vivo distribution of radioactivity was influenced by the volume of liquid that was used to instil the microsphere suspension. Comparatively large (50 microL) administration vehicle volumes resulted in substantial bronchopulmonary deposition (approximately 50% of administered dose). Intranasal instillation of microspheres suspended in 10-microL volumes tended to restrict particle deposition initially to the nasal cavity. For both administration vehicle volumes tested, the radioactivity per unit mass of excised nasal-associated lymphoid tissue (NALT) was found to be consistently elevated relative to other tissues. This corroborates the findings of other workers who have previously identified NALT as an active site of microparticle accumulation following intranasal application. Elimination via the alimentary canal was the principal fate of intranasally applied radiolabelled material. No significant concentration of radioactivity within excised gut-associated lymphoid tissue (GALT) (Peyer's patches) was noted. At latter time points we observed, in mice that received the 50-microL volume particle suspension nasally, accumulation of potentially relevant quantities of radioactivity in the liver (0.3% after 576 h) and spleen (0.04% after 576 h). Thus, our data corroborate the notion that epithelial membranes in the lung are probably less exclusive to the entry of microparticulates into systemic compartments than are those mucosae in the gastrointestinal tract or nasopharynx. This effect may contribute to the effectiveness of pulmonary delivered antigen-loaded microparticles as humoral immunogens.

Administration, Intranasal↗

Detection of DNA viruses by radioactive and non radioactive DNA probes: application to African swine fever virus.

A molecular hybridization technique using radioactive and non radioactive DNA probes, has been used to detect ASFV DNA immobilized on nitrocellulose paper. It is based on the use of plasmid pRPEL-2 as a hybridization probe. This plasmid contain the H-ClaI DNA fragment (size 5.6 Kbp) from the Spain-70 strain of ASFV. The sensitivity of detection using radioactive 32P-probes (specific activity about 2 X 10(8) cpm per microgram) was about 20 pg of viral DNA. The 32P-pRPEL-2 DNA probe can detect about 100 infected MS cells and failed to hybridize to DNA from HSV-2, MS cells or salmon sperm. The sensitivity with non radioactive probes was about 4 ng of viral DNA for a sulfonated DNA probe and 400 pg for a biotinylated DNA probe. The efficiency of DNA fixation to the filter, the effect of EDTA and of ultrasonic treatment of the sample were also investigated.

African Swine Fever Virus↗

[Resorption and incorporation of radioactive-labeled amino acids during administration of various protein carriers in rats. 3. Uptake of radioactivity by blood plasma, liver and muscular tissue and incorporation into tissue proteins after intragastric administration of 14C-leucine and 3H-glycine].

Growing male rats received diets of varying biological value (protein sources: powdered whole egg (V); fish meal (F); yeast (H); gelatine (G); protein-free diet (e)) for a 14-day feeding period. Subsequently, 14C leucine and 3H glycine were administered intragastrically. The level of uptake of 14C and 3H radioactivity into blood plasma, liver and muscular tissue and the rate of incorporation of the radioactive tracers into the proteins of these tissues was examined. A negative correlation was found to exist between the incorporation of radioactivity into liver proteins and the biological value of dietary proteins, the former being mainly dependent on the level of incorporation into the liver. For muscular proteins the rate of incorporation decreases with the decreasing biological value of the dietary proteins. This may be attributed to the fact that with poor protein nutrition the rate of protein synthesis in the skeletal muscles is also reduced. Comparative studies on the specific 14C radioactivity from free leucine made in the group on the protein-free diet and in the group receiving the whole egg diet showed that the leucine pool of the skeletal muscles was markedly redueced in animals fed a protein-deficient diet while the leucine pool in the liver remained comparatively constant.

Animal Nutritional Physiological Phenomena↗

Radioactive metal complexes with affinity for tumors. II. Biodistribution of radioactivity in cellular and subcellular fractions of tumor tissues.

The 99mTc and 57Co complexes of ethylenediamine-N,N-diacetic acid (EDDA) are accumulated in tumor tissue. The complexes and related radioactive compounds were administered to experimental animals bearing Ehrlich tumor, and the blood, tumor, abscess, and other tissues were separated, fractionated and analyzed. In blood, the EDDA complexes of 99mTc and 57Co were in dialyzable forms, whereas other tumor-nonlocalizing compounds were in undialyzable or protein-bound forms. The tumor/blood and tumor/muscle ratios of the radioactivity showed that the complexes had the high affinity for tumor tissues. Density gradient centrifugation analysis of the ascites tumor tissues showed that a significant amount of the radioactivity of the complexes was present in tumor cells. Subcellular fractionation of solid tumor tissue showed that the radioactivity was present in nuclear fraction.

Animals↗

Comparison of a radioactive and non-radioactive method for sequencing foot and mouth disease virus isolates.

The authors compare the radioactive method of detecting foot and mouth disease virus sequence products with a non-radioactive, silver stain sequencing method. The latter was found to compare favourably to the radioactive technique for detecting such products. The silver stain sequencing method was simple and did not require expensive specialised equipment. This new approach will be particularly useful in developing countries, since the method does not depend on the availability of fresh radioactive isotopes and is also safer and considerably cheaper.

Animals↗

Separation of critical radioactive and non-radioactive species from aqueous waste streams.

The separation of radioactive and non-radioactive species from the simulated DOE neutralized current acid waste (NCAW) stream was studied. Cation and anion species were referred to their possible basic compounds, and divided into seven groups (nitrate, phosphate, sulfate, fluoride, nitrite, carbonate, and hydroxide). The nitrate group (the major anion in the DOE waste streams) contains several cations species, while the rest of the groups are only in the form of sodium. The precipitation measurements were conducted in three experimental stages. In the first stage, the precipitation of sodium sulfate, sodium phosphate, sodium-sulfate-phosphate, and aluminum nitrate systems were studied using isopropylamine (IPA) as a precipitation solvent. The objectives of this stage were to evaluate the precipitation ability of IPA in precipitating these compounds individually, and to validate the consistency of the analytical instruments and the employed experimental procedure. Tests performed on the acquired data indicated a high level of experimental consistency. The removal of phosphate, sulfate and aluminum were very high. In the second stage, the precipitation studies were conducted on the: (1) nitrate group alone; (2) binary groups containing the groups of nitrate-phosphate, nitrate-sulfate, nitrate-fluoride, nitrate-nitrite, and nitrate-carbonate; (3) combined nitrate, phosphate, sulfate, and fluoride groups and (4) combined nitrate, phosphate, sulfate, fluoride, nitrite, and carbonate groups. IPA was used as a precipitation solvent. The objectives of this stage were to evaluate the interactions of these groups in the absence of the hydroxide group (e.g. DOE acid-dissolved sludge and acidified supernate streams), and the influence of such interactions on the individual removal of the targeted species. The removal of the aluminum, phosphate, fluoride, and alkaline cations were significantly high (reached 99.9%). The removal of sulfate were moderately high (reached 87%), and the removal of nitrate and alkali cations including cesium were to some extent low (reached about 50%). In the third stage, the precipitation of inorganic species from the simulated NCAW stream was studied using IPA and ethylamine (EA). The precipitation process is very feasible for reducing the radioactivity contents of alkaline cations. However, the process is less effective in separating alkali cations including cesium. The removal of polyvalent transition metals such as aluminum ion is negatively influenced by the significant presence of hydroxide. While the process is effectively capable of separating phosphate, fluoride, and sulfate, it is significantly less effective in separating nitrate and nitrite. A previously derived thermodynamics framework was used to model the precipitation measurements. The framework provided two predictive equations (the 2-Suffix and 3-Suffix equations). Both equations were reasonably adequate for predicting the solubility phase behavior of tested inorganic species in a mixed-solvents mixture as well as for estimating optimum interaction parameters. However, the 3-Suffix equation was better than the 2-Suffix equation. The parameters were useful for estimating the: (1) precipitation fractions (%P) of the studied species, for instance, at different concentration levels of similar targeted species, or in different waste streams with similar or approximate abundance of species, or at different solvents volume ratio (V(r)) where no experimental data are available.

Chemical Precipitation↗

[Influence of the radioactive strontium (89Sr) using for nuclear medical radiation therapy upon radioactive draining-water system].

PURPOSE: Strontium-89 chloride (89Sr) is a new radiopharmaceutical that provides effective pain relief for metastatic bone lesions, and is expected to be available soon in the palliative management for metastatic bone pain in Japan. Because of relatively long physical half life (50.5 days), 89Sr may affect to the radioactive draining-water system by exceeding the limits of activity concentration for radioactive drain. In this article, the influence of 89Sr use on the radioactive drainage system was simulated. METHODS: The standard tank capacity of drainage and draining frequency was determined from the results of questionnaire carried out for the nationwide medical and research institutes where radioisotope treatment are performed. On the assumption that 89Sr of 148 MBq for one therapy was used twice a week and several common radionuclides were used as the same activity as used at Chiba Cancer Center, the influence of 89Sr was estimated. The calculation was performed using the activity contamination ration into the draining-water system of each radionuclide of 0.01, which was legally determined. RESULTS: The simulation revealed that the sum of the contamination ratios of individual radionuclides exceeded a legal value of 1.0 in standard drainage with the capacity of 5 m3 and 10 m3 and draining frequency of 7 times per year. The actual contamination ratios of common radiopharmaceuticals measured at Chiba Cancer Center ranged from 1/100 to 1/1000 of the legal values. CONCLUSION: It is necessary that the legal value of activity contamination ratios into the draining-water system should be reassessed before starting 89Sr therapy.

Bone Neoplasms↗

Co-expression of cholecystokinin mRNA and tyrosine hydroxylase mRNA in populations of rat substantia nigra cells; a study using a combined radioactive and non-radioactive in situ hybridization procedure.

Improvements in the sensitivity of non-radioactive in situ hybridization histochemistry methods for detection of mRNA now make it feasible to combine the use of non-radioactive and radioactive in situ methods to visualize two mRNAs on the same tissue section. The method reported here allows the simultaneous detection of two mRNAs in one cell and therefore is ideally suited to the studies of co-expression. Here we demonstrate the co-expression of tyrosine hydroxylase (TH) mRNA and cholecystokinin (CCK) mRNA in the ventral mesencephalic dopaminergic neurones of the rat. The distribution of dopaminergic neurones containing both TH and CCK transcripts suggests, on the basis of earlier anatomical studies that these CCK/TH-containing doubled-labelled cells project mainly to the striatal matrix. Dopamine neurones believed to project to the patch compartment did not contain CCK mRNA.

Animals↗

Distribution patterns of natural radioactivity and delineation of anomalous radioactive zones using in situ radiation observations in Southern Tamil Nadu, India.

In situ radiation measurements in the beach sectors and adjacent hinterlands and along rivers in the interiors of southern peninsular India were carried out using a portable radiation survey meter. A very high intrinsic anomalous radioactivity >26microGy/h has been observed in the hinterlands within a fresh quarry and weathered boulders in the syenite rock body around Puttetti in the western Kanyakumari district of southern Tamil Nadu. Over the weathered hillocks in the hinterlands adjacent to the coast around Inayam, Kurumpanai and Midalam, the in situ radiation measurements have also exhibited high radioactivity ranging from 4 to 22microGy/h which is significantly higher than the radiation exposure rates (RER) observed along the beach sectors at various locations from Chavara to Tuticorin (1-14microGy/h). The observed radiation levels are presumably the highest concentration in southern India and it is the first time that such a high intrinsic radiogenic source in the hinterlands is reported in southwest coast of India. It is also observed based on the laboratory analysis of samples and in situ radiation data that the rivers/channels in this region contain insignificant level of radioactivity concentration and hence they do not contribute much to the placer deposits on the beaches. The placer deposits associated with significant RER (both in situ observations as well as laboratory estimates from samples) in the beach sectors from Kadiapattanam to Inayam are inferred to be derived through the country rocks/weathered hillocks in the immediate hinterlands.

Geography↗

Detection of chromosome aberrations in the human interphase nucleus by visualization of specific target DNAs with radioactive and non-radioactive in situ hybridization techniques: diagnosis of trisomy 18 with probe L1.84.

The localization of chromosome 18 in human interphase nuclei is demonstrated by use of radioactive and non-radioactive in situ hybridization techniques with a DNA clone designated L1.84. This clone represents a distinct subpopulation of the repetitive human alphoid DNA family, located in the centric region of chromosome 18. Under stringent hybridization conditions hybridization of L1.84 is restricted to chromosome 18 and reflects the number of these chromosomes present in the nuclei, namely, two in normal diploid human cells and three in nuclei from cells with trisomy 18. Under conditions of low stringency, cross-hybridization with other subpopulations of the alphoid DNA family occurs in the centromeric regions of the whole chromosome complement, and numerous hybridization sites are detected over interphase nuclei. Detection of chromosome-specific target DNAs by non-radioactive in situ hybridization with appropriate DNA probes cloned from individual chromosomal subregions presents a rapid means of identifying directly numerical or even structural chromosome aberrations in the interphase nucleus. Present limitations and future applications of interphase cytogenetics are discussed.

Cell Nucleus↗

Radioactive methionine: determination, and distribution of radioactivity in the sulfur, methyl and 4-carbon moieties.

A simple and inexpensive method is described for isolation and determination of [14C]methionine in the non-protein fraction of tissues extensively labeled with 14C. The effectiveness of the method was demonstrated by isolation of non-protein [14C]methionine (as the carboxymethylsulfonium salt) of proven radiopurity from the plant Lemna which had been grown for a number of generations on [U-14C]sucrose and contained a 2000-fold excess of 14C in undefined non-protein compounds. To our knowledge, this is the first reported assay for radioactive methionine under these demanding conditions. This method also offers an attractive alternative to the use of more expensive and sophisticated equipment for assay of radioactive methionine under less demanding conditions. An advantage is that the isolated methioninecarboxymethylsulfonium salt is readily degraded to permit separate determination of radioactivity in the 4-carbon, methyl and sulfur moieties of methionine. During this work, a facile labilization of 3H attached to the (carboxy)methylene carbon of methioninecarboxymethylsulfonium salt was observed. This labilization is ascribed to formation of a sulfur ylid.

Carbon↗

Use of unabsorbed radioactive marker substances in a re-assessment of the radioactive triolein test of fat absorption.

The radioactive triolein test has been compared with two chemical methods for the assessment of fat absorption in a group of inpatients from general medical wards. The special feature of the investigation is the use of a radioactive faecal marker to measure the completeness of faecal collections during the investigation period. Estimations of unabsorbed (131)I triolein and faecal fat measured by the chemical methods were made on the same specimens. The results showed that only 48% of the patients provided a complete collection of faeces (defined as a recovery of more than 90% of the faecal marker). From these patients good correlations were found between the triolein absorption test and the two chemical methods.It is suggested that previous criticism of the radioactive triolein test may have been due to incomplete faecal collections causing false results.

Chemistry Techniques, Analytical↗

[Nucleic acid hybridization for direct detection of Chlamydia trachomatis. Comparison of a radioactive with a non-radioactive procedure].

A total of 234 urogenital specimens were investigated with a radioactive and a nonradioactive rRNA:cDNA-hybridization test for the direct detection of Chlamydia trachomatis (Gen-Probe, San Diego, Calif.). The results were compared with those yielded by the conventional cell culture technique (McCoy cells). Using cell culture a urogenital C. trachomatis infection could be detected in 37 of 234 patients (15.8%) at two outpatient sexually transmitted disease clinics. Referred to the detection rate, cell culture sensitivity was 83.8% for the radioactive test and 86.5% for the nonradioactive DNA-hybridization test. The specificity of the radioactive test was 94.8% and that of the nonradioactive test, 93.4%.

Cells, Cultured↗

[Content of radioactive cesium in selected food products. Radioactive cesium in daily food rations of selected population groups].

The content of radioactive caesium isotopes emitting beta radiation was studied in daily food rations analysed in diets of working-class and non-working-class families from food products from the regions of Olsztyn, Poznań, Lublin, Warsaw and Wrocław in 1987 and 1988. In 1987 the highest level of radioactive caesium was found in the food rations in Olsztyn, and the lowest in the rations in Poznań (3.32 and 0.65 Bq/kg respectively). In 1988 higher radio-caesium content was found in rations composed according to the data on the diet consumed daily in non-working-class families. In that case the highest content was in the daily food ration composed in Warsaw--2.35 Bq/kg, and lowest in Poznań--1.19 Bq/kg. In the daily food rations of working-class families about one half of that value was found. The calculated mean values of both analysed rations were: 1.35 for Olsztyn, 0.89 for Poznań, and 1.86 Bq/kg for Warsaw. The calculated mean value of the contamination with radioactive caesium was in 1988 0.93 Bq/kg for the rations in working-class families (in 1987 it was 1.80 Bq/kg).

Cesium Radioisotopes↗

[Draining of radioactive waste water from departments working with radioactive material after introduction of the law for radiation protection (author's transl)].

In order to establish the drainage of radioactive waste water 1976 in the Federal Republic of Germany a new law for radiation protection was introduced. In this paper a concept is introduced how to determine specifications for constructing a decay plant using the given data of a department for nuclear medicine. The given boundary conditions were to secure a low concentration of radioactive material in order to establish the drainage of extreme low activities without diluting it.

Berlin↗