Sfpi1/PU.1 mutations in mouse radiation-induced acute myeloid leukaemias affect mRNA and protein abundance and associate with disrupted transcription.

Radiation-induced acute myeloid leukaemias (AMLs) in mice are characterised by deletions and point mutations in the Sfpi1/PU.1 transcription factor. Six AML cell lines were used to examine the impact of three previously described R235 point mutations. AML cells carry myeloid and stem cell markers and the R235 mutations differentially affect mRNA and protein abundance. Expression of Sfpi1/PU.1 target genes was deregulated in a broadly similar fashion irrespective of R235 mutation including Flt3, which is frequently subject to activating mutations in human myeloid leukaemias. While R235 mutations differentially affect protein abundance they resulted in similar disruption of Sfpi1/PU.1 functions.

Investigations of the potential bioavailability of 210Po in some foodstuffs.

Extraction of 210Po from lamb's liver, pig's kidneys, mussel flesh and brown crabmeat with a series of solvent systems has been used to gain some insight into the uncertainty about the gastrointestinal absorption factor for Po incorporated into foodstuffs. By extracting the tissues with diethyldithiocarbamate dissolved in chloroform, and also in methanol, it has been shown that 210Po is more effectively extracted from lamb's liver and crabmeat than it is from each of the other tissues. A similar pattern of extractability is also evident for aqueous solutions of citric acid on crabmeat and mussel flesh. Of particular note is the low extractability by dimethylsulphoxide (3%) of 210Po in mussel flesh as opposed to 24% extractability of 210Po from crabmeat. 210Po-binding macromolecules of about 10 kDa have been released from crabmeat and mussel flesh by digestion with pepsin.

Effects of macromolecular chelating agents on the release of 239Pu and 59Fe from rat alveolar macrophages after phagocytic uptake of 239Pu-59Fe-iron hydroxide colloid.

The chelation-mediated mobilization of 239Pu and 59Fe from rat alveolar macrophages which had engulfed 239Pu-59Fe-hydroxide colloids was studied by exposing the macrophages to diethylenetriaminepentaacetic acid and macromolecular weight forms of diethylenetriaminepentaacetic acid. The release of 239Pu and 59Fe by these macromolecular weight chelating agents and diethylenetriaminepentaacetic acid was assessed in monolayer culture and shown to be dependent upon the nature of the engulfed colloids. Whereas calcium-diethylenetriaminepentaacetic acid, aminoethylcellulose-diethylenetriaminepentaacetic acid, and aminoethylcellulose-diethylenetriaminepentaacetic acid oxidized with periodic acid (aminoethylcellulose-diethylenetriaminepentaacetic acid(I2) exhibited some effectiveness at releasing 239Pu and 59Fe from macrophages which had engulfed the low aggregative colloid, they were less effective in the release of these radionuclides from the high aggregative colloid. The efficacy of these chelating agents on the removal of radionuclides from alveolar macrophages differed between iron and plutonium, suggesting the significant difference in the dissolution and subsequent metabolism of these radionuclides in alveolar macrophages. From the similarities of the percentage increases in 239Pu mobilized from the low aggregate colloid by aminoethylcellulose-diethylenetriaminepentaacetic acid (I2) and diethylenetriaminepentaacetic acid, it would appear that both forms of diethylenetriaminepentaacetic acid could be accessing the same deposits of 239Pu; however, for 59Fe the percentage increases are quite different for the two forms of diethylenetriaminepentaacetic acid. SiO2-diethylenetriaminepentaacetic acid, an insoluble macromolecular weight chelating agent, was ineffective at releases 239Pu but was slightly effective at releasing radioiron.

Chelating agents and the regulation of metal ions.

Up to about the early 1980s it was perhaps still possible to summarize in a review of a moderate length the development of the medicinal applications of chelation chemistry and the exploitation of such chemistry in regulating the metal ion concentrations in the body. However, in the last few years there has a great surge in the development of chelation chemistry and its usage in medicine and related areas of life sciences research. It is no longer the case that such a review primarily concentrates upon the use of chelating agents in removing toxic metals from the body but it must now cover the use of chelating agents in the imaging procedures nuclear medicine and magnetic resonance imaging (MRI), the use of chelating agents in unravelling the biochemistry of reactive oxidative species (ROS) and the control and measurement of intracellular calcium ions. It is in the recent applications that there have been the greatest developments over the last ten years.

Speciation of plutonium in potato and the gastrointestinal transfer of plutonium and americium from potato.

Complexation of Pu(IV) and Am(III) by naturally occurring agents such as citrate and phytate might enhance their uptake from the digestive tract. However, the extent to which they enhance the gastrointestinal uptake of actinides from foodstuffs is far from resolved, as this study shows. Investigations of the chemical forms of Pu(IV) and Am(III), by gel permeation chromatography, in simulated digests of potato tubers naturally radiolabelled with 239Pu(IV) and 241Am(III) have shown that neither citrate nor phytate appear to determine their chemical forms. Therefore, it is possible that these are not the complexing anions which determine the gastrointestinal transfer of these radioelements from potato meal. Isotachophoretic analyses of the juices pressed from tubers and solutions prepared by simulated digestion of potato tubers have demonstrated the presence of several low molecular weight anions. These anions might be complexing agents because they possess an isotachophoretic mobility similar to that of citrate; some of these anions remain unidentified. Whereas 239Pu and 241Am were used in the foregoing studies, 238Pu and 241Am were used to produce either in vitro or naturally radiolabelled potatoes for gastrointestinal transfer measurements using rats and hamsters. Gastrointestinal transfer values of 0.13 +/- 0.05% (mean +/- standard error of mean) and 0.16 +/- 0.06% were determined with rats for the uptake of 238Pu and 241Am, respectively, from naturally labelled potato meal. Higher gastrointestinal transfer values were obtained for hamsters: for 238Pu and 241Am the transfer values from naturally labelled meal were 0.25 +/- 0.08% and 0.33 +/- 0.07%, respectively. Similar values were observed for uptake from in vitro labelled potato meal. On the basis of the similarity of the values for the naturally labelled potato meal and for the 'spiked' potato meal it would appear that biological incorporation is not necessary for the binding of the actinides to the ligands which will determine gastrointestinal transfer.

Use of paramagnetic chelated metal derivatives of polysaccharides and spin-labeled polysaccharides as contrast agents in magnetic resonance imaging.

Soluble and insoluble polysaccharides were derivatized with diethylenetriaminepentaacetic acid (DTPA) and/or spin-labeled with 2,2,6,6-tetramethylpiperidine-1-oxyl (TEMPO). Polysaccharides derivatized with DTPA were prepared via cyanogen bromide activation, coupling to a diamine linker, and to DTPA anhydride. Spin-labeled polysaccharides were also prepared via cyanogen bromide activation. The extent of derivatization for dextran (18 kDa) was about 120 glucose units per DTPA, and for cellulose and starch about 15-30 units per DTPA. For spin-labeled polysaccharides, the average loading ranged from 1 nitroxide per 16 glucose units for starch to 181 for dextran (82 kDa). These derivatized paramagnetic polysaccharides were shown to be more effective relaxants than the small paramagnetic molecules alone. Both soluble and insoluble polysaccharide-linker-DTPA-Gd(III) complexes were effectively cleared from the body (rats) after oral administration. After intravenous administration, the biodistribution of dextran-linker-DTPA-Gd(III) complexes differed significantly from that of GdDTPA. Reduction of the nitroxide by ascorbic acid was retarded in the polysaccharide derivatives, particularly in starch derivatized with both nitroxide and linker-DTPA-Cu(II). These agents showed contrast enhancement in the gastrointestinal tract of rabbits.

Effects of macromolecular chelators on intestinal cadmium absorption in mice.

Suppression of absorption by macromolecular chelators have been successful with several metals. In this paper a series of immobilized chelators ranging from DTPA to S-containing soft bases have been synthetized and investigated for ability to suppress intestinal uptake of 109Cd2+ in mice. Dextran-0-ethyl-mercaptan, xanthates derived from polysaccharides and polyvinyl alcohol, dithiocarbamates of polyethylene imine and aminoethyl cellulose, and DTPA immobilized on aminopropyl silica were all ineffective. DTPA immobilized on aminoethyl cellulose even enhanced the intestinal uptake. The macromolecular chelators were without extensive effect on organ distribution of absorbed cadmium, except for dithiocarbamate immobilized on polyethylene imine, which enhanced the deposition of cadmium in several organs including the brain. Although the results are discouraging, they indicate that design and synthesis of immobilized vicinal dithio compounds may represent an avenue for development of non-absorbable chelators with high affinity for cadmium.

Effect of chelating agent structure on the mobilization of cadmium from intracellular deposits.

An examination of the efficacy of several structural types of chelating agents in the removal of cadmium from its intracellular deposits in mouse liver and kidneys reveals that of the structural types examined, only dithiocarbamates and a vicinal dithiol were able to mobilize cadmium from such intracellular sites. Esters of L-cysteine, a macrocyclic thioether, and a disulfide of a dithiocarbamate were unable to cause any appreciable decrease in either renal or hepatic cadmium levels. Charged groups such as carboxylic acid groups reduce the efficacy as well as the toxicity of the structural types that can otherwise mobilize such cadmium. It was also found that the administration of a cadmium-binding polymer ip leads to only a very slight net excretion of cadmium, while the po administration of this polymer leads to no net additional cadmium excretion. Of the compounds newly reported here, some are approximately equal in cadmium-mobilizing efficacy to the most effective of previously reported compounds.

The retention of 14C-DTPA in human volunteers after inhalation or intravenous injection.

This study in human volunteers was designed to compare the retention of diethylenetriaminepentaacetic acid (DTPA) in the body after intravenous (i.v.) injection with that following inhalation by using a 14C labelled tracer. After i.v. injection retention in the blood could be described by three exponential components with half-times of about 1.4 min (approximately 60%) 14.3 min (approximately 20%) and 95 min (approximately 20%). By 24 hr more than 99% of the 14C-DTPA had been excreted in the urine and less than 0.5% remained in the plasma. After inhalation of 14C-DTPA retention in the lungs could be represented by a single component with a half time of about 75 min. As a consequence the length of time that a therapeutically useful amount of DTPA is retained in the body is approximately twice that following intravenous injection.

Actinide transport across cell membranes.

Protactinium uptake into the normal liver does not exceed 3%, but when the phospholipid levels in the liver are elevated by administration of thioacetamide this uptake increases to 31%. Phosphatidic acid, which is absent from the normal liver, has been shown to extract protactinium into organic solvents. However, phosphatidylserine, a component of normal liver cell membranes, does not extract protactinium. It might be conjectured that this is why so little protactinium is taken up by the normal liver. The hypothesis is advanced that phosphatidylserine, which is known to complex plutonium, americium and curium, may regulate the uptake of these elements by liver.