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Excretion of radioactivity in faeces and urine of rats injected with 3H,14C-lipopolysaccharide.

The route of excretion of lipopolysaccharide (LPS) and its possible degradation in vivo was studied in rats using biosynthetically radiolabelled LPS from Salmonella abortus equi, carrying 3H activity exclusively in fatty acids and 14C activity in fatty acids and sugars. Following intravenous injection of the above LPS in AS2 rats with or without anaesthesia, excretion of radioactivity occurred mainly in the faeces and to smaller extent in urine. The rate of excretion was slow, a large part of the radioactivity being still present in the liver after 14 days. In faeces the percent recovery of 3H (18%) was lower than that of 14C (32%) suggesting loss of tritium activity and thereby of fatty acids from the excreted LPS. A similar loss of tritium was also found in the material remaining in the liver and spleen 14 days after LPS administration. In urine the material recovered during 14 days (about 7% of injected) was different from the original LPS, 70% of 14C activity being dialysable and practically all 3H activity being volatile. Similar results were also obtained following administration of the LPS intraperitoneally under anaesthesia. However, when the LPS was administered intraperitoneally without anaesthesia, in the majority of the animals, 90% of 14C and 54% of 3H was excreted in faeces within 3 days, suggesting that both route of administration and use of anaesthesia during injection influence the subsequent rate of excretion of LPS.

Animals↗

[Delayed effects of acute radiation injuries by radioactive carbon (14C-glycine)].

A study was made of late biological effects of radioactive carbon (14C-glycine) applied in acute doses. The course of radiation sickness, at its acute stage, is similar to that of acute radiation sickness caused by external gamma-irradiation. The recovery is slow, and the disease becomes chronic exhibiting metabolism disturbances and early death of animals. Radioactive carbon has a pronounced blastomogenic action.

Animals↗

Experimental study of measurement of regional bone blood flow in the adult mongrel dog using radioactive microspheres.

To date, no one has conclusively accurately measured the regional bone blood flow mainly due to the lack of a proper investigative method. In this study, regional blood flow in the femur and tibia in adult mongrel dogs were studied using 85Sr-and 141Ce-labeled microspheres with a diameter of 15 +/- 5 microns. The blood flow rates were calculated by relating the radioactivity from tissue samples that from sample blood with a known flow. The highest rate of blood flow, 48.7 ml/min/100 g, was found in the femoral marrow and the lowest, 3.0 ml/min/100 g, in the diaphyseal cortex of the tibia. Flow rates in the metaphyseal regions, about 15 or 20 ml/min/100 g, were one and a half or 2 times greater than those in the epiphyseal regions. These values were mostly within the range of values reported by other investigators. Radioactive microspheres can be applied not only to physiological conditions but also to various pathological conditions if their size and the dose, the site and method of injection and tissue sampling are investigated further.

Animals↗

[Incorporation of radioactively labelled phosphoglycerides into the retina after subcutaneous application (author's transl)].

Radioactively labelled phosphoglycerides were given subcutaneously to rabbits and the incorporation of these substances into the retina and other organs has been studied. In a first examination the phospholipids were labelled with 3H-oleic acid, in a second examination we used 32P for labelling. We could show, that both the fatty acids and the polar head group of the phosphoglycerides were incorporated into the retina lipids. The specific radioactivity of the retina phosphoglycerides was always higher than that of lipids extracted from other organs.

Animals↗

Assay of nanogram levels of triglyceride lipase with a radioactive substrate.

A simple, sensitive procedure for the determination of triglyceride lipase activity has been developed. Nanogram amounts of oleic acid hydrolyzed from commercially available [(14)C]triolein were readily determined by the counting of the radioactivity of substrate and product after their rapid chromatographic separation on copper hydroxide-impregnated ion-exchange paper. Comparison of the relative amounts of radioactivity of the separated substrate and product gave an estimate of the percentage of hydrolysis of substrate. Comparison of results with a standard of pure lipase enables one to express the amount of hydrolysis in terms of the standard lipase. The results show that measured activity is a linear function of time up to 1 hr of incubation and of amounts of enzyme up to 125 ng. Reproducibility of the test is good.

Carbon Isotopes↗

[Radioactive labeling of Blatella germanica].

For ecological investigations in natural populations of Blatella germanica a new method of dry incorporation of a porcelain tracer is developed. The quality of this method in contrast with other non radioactive and at present radioactive methods depends on the stable position of the tracer and the possibility of taking different marking substances for individually marking a large number of experimental animals. The identification bases on the previously investigated half-life values. The new method in combination with the recaptivate method is a good possibility to analyse a population of cockroaches.

Animals↗

Determination of the specific radioactivity of fatty acids separated as their methyl esters by gas-liquid chromatography.

Free or combined (3)H-labeled fatty acids are converted to their methyl-(14)C esters or, if labeled with (14)C, to their methyl-(3)H esters. For a given specific radioactivity of the methyl group, the nuclide ratio in the esters separated by GLC is a direct measure of the specific radioactivity of the fatty acids, and quantitative collection is unnecessary. Methods of methylation with minimum quantities of labeled methanol, and of deriving nuclide ratios from channel ratios in a scintillation spectrometer, are given.

Carbon Isotopes↗

Preparation of radioactive Tay-Sachs ganglioside labeled in the sialic acid moiety.

A procedure is described for the preparation of Tay-Sachs ganglioside specifically labeled in the sialic acid portion of the molecule. Rat brain gangliosides were labeled biosynthetically by the intracranial injection of N-acetyl-(3)H-D-mannosamine. Radioactive gangliosides were isolated and selectively degraded with bacterial neuraminidase and rat liver beta-galactosidase to Tay-Sachs ganglioside-(3)H. Radioactivity in the labeled product was confined to the N-acetyl-neuraminic acid portion of the molecule.

Animals↗

[On the physiology of growth and riboflavin overproduction of Eremothecium ashbyii. III. Investigations on the incorporation of radioactive labeled substrates in cell material and riboflavin (author's transl)].

The incorporation of glycine-2-14C and adenine-U-14C in cell material and riboflavin was investigated in order to determine the proportions metabolites were channeled in growth processes and product synthesis of Eremothecium ashbyii. Extraction- and measurement methods were developed to compare the incorporation of radioactive metabolites into different cell fractions. Young cells incorporate the labeled compounds in a high rate into the cell material. With increasing age of the culture the incorporation of radioactivity is more directed to riboflavin. 96 hours old cells incorporate about seven times more 14C-adenine into riboflavin than into the cell material.

Adenine↗

[Metabolism and pharmacokinetics of piprozoline in rat, dog and man. 2nd Communication: Determination of metabolic profiles with the aid of an HPLC-radioactivity detection system following direct injection of urine and bile onto a reversed-phase column (author's transl)].

This paper describes the determination of metabolic profiles of 14C marked ethyl (Z-3-ethyl-4-oxo-5-piperidino-thiazolidin-2-ylidene) acetate (piprozoline, Gö 919, Probilin) in rat, dog and human urine and rat bile with the aid of an HPLC-radioactivity detector system following the direct injection of large quantities onto an analytical C-18 column. Metabolite 1 and two glucuronic acid conjugates of metabolite I were determined quantitatively, isolated and identified. Following HPLC separation, the specific detection of radioactivity permits the determination of metabolic profiles without prior clean-up so that unstable metabolites can also be analyzed.

Adult↗

Thyroid uptake of radioactive iodine in hyperthyroidism.

We compared the 24-hour uptake of radioactive iodine (RAIU) in hyperthyroid patients diagnosed from 1970 to 1972 (Group A) and from 1975 to 1976 (Group B). Mean RAIU values in Group B decreased from those in Group A. In 1969 we lowered the normal range for the RAIU test to 0% to 24%, and used the new range for the diagnosis of hyperthyroidism in both groups of patients. The RAIU in Group B is within normal limits in 14% of the patients with Graves' disease and 80% of the patients with toxic nodular goiter (TNG). This study demonstrates that in our geographic area, the 24-hour RAIU has become a poor test for the diagnosis of hyperthyroidism, particularly TNG. However, the test should be performed before radioactive iodine therapy because it is necessary in calculating the therapeutic dose.

Adult↗

Studies on the inhibition of intestinal absorption of radioactive strontium. V. The effect of administration of calcium alginate.

A method of selective suppression of absorption of radioactive strontium from ingested food material is described which allows calcium to be available to the body. In the present study the effects of administering calcium alginate were determined. Studies on the relative binding of Ca and Sr by calcium alginate in vivo are also important to the investigation of calcium-strontium exchange.Samples of calcium alginate were obtained commercially or prepared from sodium salts and the binding properties with Ca(45) and Sr(89) were tested in vivo. There was a free exchange of radioactive calcium as well as of strontium with bound inactive calcium. The amount of Ca(45) retained in the gut by calcium alginate is proportionally less than Sr(89), so that the ratio Sr(89)/Ca(45) in the bone is consistently and significantly lower in treated rats. Calcium alginate acts differently from other calcium salts, although in a manner similar to sodium alginate.

Alginates↗

[Problems in the use of radioactively labelled bacteria in experiments. 2. Quantitative method for the evaluation of pathogens in the calf lung by means of labelled bacteria after aerosol or intratracheal administration].

By means of two aerosol apparatures radioactive labelled Pasteurella-multocida-germs were given to calves via a respiratory mask. Labelling of Pasteurellae was done internal by using special nutritive media with Fe-59 or P-32. In a period of 10-25 minutes 5 X 10(9)-10(10) labelled bacteria were given to the animals for inhalation. Another animal group was injected intratrachealy with the same quantity of labelled germs. Immediately after termination of germ application the animals were killed and dissected. After solution or homogenization, respectively of the total lung tissue radioactivity was determined by liquid-scintillation-counting (Fe-59) or Cerenkov-measurement (P-32), respectively. The activities recovered from lung homogenates rendered is possible to calculate the received amount of bacteria. 40-80% of germs would be recovered in the lung after application by intratracheal injection while only 1-7% could be recovered after individual aerosol application lasting for 15-25 minutes.

Aerosols↗

[Characteristics of collagen preparations labelled in vitro by 14C-acetic anhydrides. Their use for the determination of collagenase activity and as radioactive markers].

A modified procedure in developed for acetylation by means of 14C-acetic anhydride of highly purified collagen I preparations. The acetylated collagen exhibited high specific radioactivity and was effectively hydrolyzed by bacterial collagenase. The 14C-acetylated collagen was used as a radioactive marker in electrophoretic analysis of labelled proteins.

Acetates↗

Glucose induced inhibition of radioactive nucleotide efflux from mouse pancreatic islets is dissociated from an increase in islet oxygen uptake.

Glucose at 1.7 and 5.6 mmol/l, pyruvate (30 mmol/l) and glutamine (10 mmol/l) all stimulated islet respiration slightly. More marked stimulation was achieved with glucose 8.3 mmol/l. When pyruvate (30 mmol/l) was added to islets respiring in the presence of different concentrations of glucose (1.7, 3.3 or 5.6 mmol/l), a stimulatory effect was only observed at the highest glucose concentration. Stimulation of islet oxygen uptake was also achieved when glucose 8.3 (but not 5.6) mmol/l was added to islets respiring in glutamine 10 mmol/l. Radioactive nucleotide efflux was, however, progressively inhibited by increasing the glucose concentration up to 5.6 mmol/l both in the absence or presence of glutamine 10 mmol/l. It is concluded that the glucose-induced inhibition of radioactive nucleotide efflux is not obligatorily linked to respiration.

Adenine Nucleotides↗

[In vitro method determining sensitivity of anticancer agents by incorporation of radioactive precursors].

A new sensitivity test of anticancer agents was developed to measure the lethal effects of cancer cells by the incorporation of radioactive precursors. The thousand cancer cells were cultured in a microplate in the presence of anticancer agents. These cells were exposed to radioactive precursors. Two or three days later, the cancer cells were harvested on a glass fiber filter by a multiple automatic cell-harvester and the incorporation of precursors was counted by a liquid scintillation counter. In this study, the in vivo results of drug testing in animal model systems were compared with drug sensitivities. Mice inoculated Ehrlich ascites cells were treated with various kinds of anticancer drugs. The development of the cells was compatible with the result of the sensitivity test. The growths of Lauson and ME-180 cells derived from human cancers implanted subcutaneously to nude mice were also well correlated with this sensitivity test.

Animals↗

[Uptake of radioactive precursors for the evaluation of sensitivity of anticancer drugs against cancer cells].

An in vitro drug sensitivity test was developed to estimate the lethal effects of drugs on cancer cells. Cancer cells we incubated in the presence of radioactive precursors and anticancer drugs. The drug effects were estimated from the changes in rates of incorporation of precursors into DNA, RNA, and protein. Handling of a large number of samples became possible using a microtiter plate and a multiple automatic cell harvester. Incorporation of radioactive precursor showed linear correlation with drug-induced lethality. The results of this test were also correlated well with in vivo regression of tumors of mice. This test appeared to be more reliable than other similar test of cell lethality in vitro. For utilization in clinical studies, a test plate was simplified and 25 human cancers were tested. The results demonstrated the generally good correlation between in vitro and in vivo tumor sensitivities.

Animals↗