Consultative committee on ionizing radiation: Impact on radionuclide metrology.

In response to the CIPM MRA, and to improve radioactivity measurements in the face of advancing technologies, the CIPM's consultative committee on ionizing radiation developed a strategic approach to the realization and validation of measurement traceability for radionuclide metrology. As a consequence, measurement institutions throughout the world have devoted no small effort to establish radionuclide metrology capabilities, supported by active quality management systems and validated through prioritized participation in international comparisons, providing a varied stakeholder community with measurement confidence.

Evaluation of Handheld Radionuclide Identifiers.

Characterization of commercially available instruments for measurement and identification of unknown radionuclides was carried out in support of the development and testing of the American National Standards Institute (ANSI) standard, N42.34, "Performance Criteria for Hand-held Instruments for the Detection and Identification of Radionuclides." Measurements were based on the performance of the devices, i.e., the capability of the detectors to ensure a correct radionuclide identification in a given time interval for various radioactive sources.

Use of resonance ionization mass spectrometry for determination of Cs ratios in solid samples.

A method to quantitatively determine the radionuclidic content of complex samples with a minimum of chemical preparation has been under development at the National Institute of Standards and Technology (NIST). Based on earlier studies with resonance ionization mass spectrometry (RIMS), a graphite furnace was added to the RIMS system at NIST. Efficiency determinations for 133Cs in solution and isotopic ratio measurements of cesium (135Cs/137Cs) for an NIST nuclear burn-up sample using RIMS and for a sediment natural matrix standard using thermal ionization mass spectrometry are presented.

Modeling energy deposition and cellular radiation effects in human bronchial epithelium by radon progeny alpha particles.

Energy deposition and cellular radiation effects arising from the interaction of single 218Po and 214Po alpha particles with basal and secretory cell nuclei were simulated for different target cell depths in the bronchial epithelium of human airway generations 2, 4, 6, and 10. To relate the random chord lengths of alpha particle tracks through spherical cell nuclei to the resulting biological endpoints, probabilities per unit track length for different cellular radiation effects as functions of LET were derived from in vitro experiments. The radiobiological data employed in the present study were inactivation and mutation (mutant frequency at the HPRT gene) in V79 Chinese hamster cells and inactivation and transformation in C3H 10T1/2 cells. Based on computed LET spectra and relative frequencies of target cells, probabilities for transformation, mutation, and cell killing in basal and secretory cells were computed for a lifetime exposure of 20 WLM. While predicted transformation probabilities were about two orders of magnitude higher than mutation probabilities, they were still about two orders of magnitude lower than inactivation probabilities. Furthermore transformation probabilities for basal cells are generally higher than those for secretory cells, and 214Po alpha particles are primarily responsible for transformations in bronchial target cells.

Biomarkers of OH radical damage in vivo.

Mechanisms of formation of o-tyrosine (o-Tyr) and thymine glycol (TG), the two possible markers of OH radical generation in biosystems and in vivo are described. The o-Tyr measurements require invasive approaches, while TG detection may be accomplished by noninvasive analysis in the urine.

Formation of ortho-tyrosine by radiation and organic solvents in chicken tissue.

Fresh chicken breast and beef incubated in water were found to contain no o-Tyr at the current levels of detection (0.1 ppm) by capillary gas chromatography/mass spectrometry and selective ion monitoring. In contrast, samples incubated at 37 degrees C in the presence of ethanol, benzene, or carbon tetrachloride (used in fat extraction) contained large quantities (2.5-5.1 ppm) of o-Tyr. No o-Tyr was detected in the water-insoluble fraction of meat treated with carbon tetrachloride after triple extraction by water. However, reaction of radiation generated .OH in gamma-irradiated fresh chicken tissue with endogenous phenylalanine yields o-Tyr with a linear yield-dose response in both water-soluble and -insoluble tissue fractions. Nonradiolytically generated .OH is suggested to be formed through a mitochondrion-mediated Haber-Weiss reaction in association with water-soluble proteins since the yields of o-Tyr in beef, a tissue with a higher mitochondrial content, are four times greater than in the chicken breast tissue.

Modification of deoxyribose-phosphate residues by extracts of ataxia telangiectasia cells.

The release of DNA 5'-terminal deoxyribose-phosphate residues from enzymatically incised apurinic/apyrimidinic sites by human cell extracts has been under investigation. During the course of these studies, we observed that ataxia telangiectasia cell extracts modify deoxyribose-phosphate (dRp) residues by converting them to an altered form, dRp-X, which shows altered chromatographic properties on HPLC analysis. The chemical nature of the adduct is as yet unknown, but dRp-X is stable to both heat and acid. The modification requires an enzymatic activity and a low-molecular weight co-factor. Extracts of normal cells contain a dialyzable inhibitor that suppresses the reaction occurring with ataxia telangiectasia cell extracts. Formation of dRp-X has been observed in 7 out of 7 ataxia telangiectasia lymphoblastoid lines which represent at least 3 genetic complementation groups. Similar modification of dRp did not occur with extracts of cells of normal origin, nor those representing Fanconi's anaemia, xeroderma pigmentosum, Bloom's syndrome, Werner's syndrome or Friedreich's ataxia.

Generation of oxy radicals in biosystems.

Many recent lines of evidence indicate that endogenous free radicals contribute to spontaneous mutagenesis through the direct induction of DNA damage. However, the mechanisms underlying this process are not yet fully understood. A brief overview of the knowledge that is currently available is provided here, with emphasis on the generation of oxy radicals in biosystems, the reactions of those radicals with biomolecules, and the induction of oxidative DNA base damage that might lead to mutation.

Mechanisms of free radical chemistry and biochemistry of benzene.

omicron-Tyrosine (omicron-Tyr) was used as a specific biomarker for .OH radicals generated in biosystems. Specificity of omicron-Tyr as an .OH biomarker was based on previous studies in systems exposed to ionizing radiations. Fresh muscle tissue incubated with benzene for 1 hr at 38 degrees C exhibits formation of omicron-Tyr as seen in the cases of ethanol- and carbon tetrachloride-exposed systems. Gas chromatography/mass spectrometry selective ion monitoring measurements of omicron-Tyr yields in chicken breast muscle incubated with water or benzene indicate levels of less than 0.1 ppm and 3.0 +/- 0.5 ppm of omicron-Tyr, respectively. Formation of .OH is presumed to originate via a Haber-Weiss reaction of H2O2 with Fe(II) preceded by the formation of .O2- and H2O2 from distorted mitochondria.

Intramolecular H atom abstraction from the sugar moiety by thymine radicals in oligo- and polydeoxynucleotides.

Hydroxyl radical addition to uracil (U) has been suggested to lead to strand breaks in polyuridylic acid, an occurrence attributed in part to H atom abstraction by .U-OH radicals from the ribose moiety [D.G.E. Lemaire et al., Int. J. Radiat. Biol. 45, 351-358 (1984)]. We have investigated this particular reaction by means of the hydroxyl radical-induced products of thymine (T), pT, TpT, TpTpT, polythymidylic acid (poly-T), (T + dR) poly-dA.poly-T, and a mixture of T and 2-deoxyribose (dR). The major monomeric product of .T-OH in TpT, TpTpT, poly-T, and poly-dA.poly-T was found to be 5-hydroxy-6-hydrothymine (H-T-OH), while that in T, pT, and T plus dR was thymine glycol (HO-T-OH). These results indicated that the intramolecular H atom abstraction from a nearby sugar (in this case, deoxyribose) moiety by base radicals, i.e., .T-OH, occurs in oligo- and polydeoxynucleotides of T. In poly-T, the yield of H-T-OH is not much greater than in TpT or TpTpT, indicating that the abstraction of an H atom from the sugar moiety of a nucleotide subunit further than two nucleotides along the chain may not be significant. Additionally, a corresponding decrease in the yield of HO-T-OH with an increase in the yield of H-T-OH suggests that the formations of these two types of thymine products are competitive.

Free radical-induced cross-linking of polydeoxythymidylic acid in deoxygenated aqueous solution.

Radiation-generated hydroxyl radicals and hydrogen atoms were shown to induce the cross-linking of polydeoxythymidylic acid (mol X wt approximately 170 000) in N2O-saturated acqueous solution. The irradiated samples were hydrolysed with formic acid and then analysed by high performance liquid chromatography. Products were isolated and subsequently characterized by capillary gas chromatography-mass spectrometry. The presence of previously described monomeric thymine products was also shown. Yields were determined and mechanisms of formation were described for the products.

OH radical-induced products of tyrosine peptides.

Reactions of radiation-generated OH radicals with tyrosine and its homopeptides, i.e. L-Tyr-L-Tyr and L-Tyr-L-Tyr-L-Tyr, in N2O saturated solutions were shown to give crosslinks between the peptide chains with high yields. High-performance liquid chromatography, capillary gas chromatography and mass spectrometry were used for isolation and identification of the monomeric and dimeric products. Evidence is presented for the crosslinking to occur through C-C and C-O-C bonds. To the best of our knowledge, the formation of the ether type of crosslink is demonstrated for the first time. Mechanisms of product formation are also discussed, which involve radicals that were described in previous pulse radiolysis studies.