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PubMed1999

A rapid and sensitive quantitative kinase activity assay using a convenient 96-well format.

Activation of protein kinases in response to growth factor and extracellular matrix stimulation has been implicated in regulating a number of cell functions including differentiation, gene expression, migration, and proliferation. An improved quantitative assay for measuring protein kinase activity is crucial to the detailed study of this important category of signaling proteins and their role in regulating cell behavior. We describe a modified in vitro kinase activity assay that is both sensitive and quantitative. It offers several advantages when compared to the traditional immunoprecipitation/kinase assay: (i) high sensitivity that reduces the required amount of cell lysate by an order of magnitude, (ii) an immunoseparation technique utilizing antibody immobilization onto the surface of microtiter wells that replaces the cumbersome immunoprecipitation method, (iii) a 96-well plate configuration that eases handling of multiple samples and increases throughput of the assay, and (iv) the use of 96-well filter plates that greatly reduces radioactive liquid waste generation. While we implement this technique in a case study for measuring the activity of extracellular signal-regulated kinase 2 (ERK2), this assay can be extended to studying other protein kinases by using an appropriate antibody and in vitro substrate for the kinase of interest.

Animals↗
PubMed1981

Residual activity of Tc-generators.

Owing to contamination by long-lived radio-isotopes, the residual activity of an old 99Mo-99mTc generator may be large enough to prevent its disposal. The long-lived impurities of one generator were determined by energy spectrum and half-life measurements. In this generator 46Sc, 51Cr, 60 Co, 92mNb, 95Nb and 124Sb nuclides were found. Only the half-life of 60Co exceeds 3 months, thus a moderately long storage of the generator reduces the activity, and its disposal is then safe.

Radioactive Waste↗
PubMed1991

Approaches to investigating clusters: by example, childhood leukaemia near Sellafield.

There is occasionally concern about suspected clusters of disease in geographically localised areas among members of the general public or other groups. These occurrences need a systematic approach to their investigation. Such an approach needs to consider the definition and characteristics of the cluster and of the putative environmental cause for concern. Depending on the outcome of these considerations there will be different actions, possibly involving epidemiological studies. The necessary components of the approach and resulting actions are outlined in this paper, and illustrated by the example of childhood leukaemia rates around the Sellafield nuclear plant.

Case-Control Studies↗
PubMed1991

Components of total irradiation dose in Switzerland and their ranges.

The present knowledge about dose components in Switzerland and their ranges is reviewed. Considerable ranges are found for doses induced by radon decay-products and by cosmic and terrestrial radiation. Yearly doses from radon decay-products show average values between about 1 and 20 mSv in different communities and individual values up to about 150 mSv. The reliability of these average values is, however, limited, because radon concentrations have been measured up to now only in a small number of houses, and because corrections of the raw data are necessary, increasing the uncertainty of the results. Doses from terrestrial and cosmic radiation show locally variable values between about 0.5 and 1.5 mSv per year. These doses are mainly derived from outdoor measurements. Therefore, these results also are only of limited use in possible epidemiological applications.

Background Radiation↗
PubMed2002

Lesson learned from Co-60 accident in Thailand.

The causes and consequences of a Co-60 radiation accident in Samutprakarn Province, Thailand, were scrutinized to learn lessons aimed at preventing future radiation accidents. "Orphan sources" may end up in scrapyards. An out-of-use Co-60 medical teletherapy source, left unattended in a disused parking area belonging to a Medical Dealer, was stolen and sold to a scrap dealer in Samutprakarn Province at the end of January 2000. Because of its valuable appearance, a number of workers in the scrap trade who were not aware of radiation hazards managed to dismantle all parts. The Co-60 source was removed and left unshielded among pieces of scrap metal in the yard of the scrap shop. Some workers immediately became sick. Eighteen days later when they went to a local hospital their symptoms were recognized as radiation sickness and the incident was reported to the Office of Atomic Energy for Peace (OAEP) in Thailand. The unshielded source, with an estimated activity of 15.7 TBq (425 Ci), was retrieved soon after by an emergency team and placed in safe storage at the OAEP premises. Ten victims developed radiation sickness symptoms, of which three died soon after the accident. The accident alarmed the public, and has raised national concerns. The accident is similar in some ways to the 1987 radiation accident at Goiania, Brazil, involving a Cs-137 radiotherapy source. If not properly disposed of orphan radiation sources can lead to serious injury or even death. The accident highlights the need for security of spent high activity sources and the importance of regulatory controls.

Cobalt Radioisotopes↗
PubMed1998

Microbiology of spent nuclear fuel storage basins.

Microbiological studies of spent nuclear fuel storage basins at Savannah River Site (SRS) were performed as a preliminary step to elucidate the potential for microbial-influenced corrosion (MIC) in these facilities. Total direct counts and culturable counts performed during a 2-year period indicated microbial densities of 10(4) to 10(7) cells/ml in water samples and on submerged metal coupons collected from these basins. Bacterial communities present in the basin transformed between 15% and 89% of the compounds present in Biologtrade mark plates. Additionally, the presence of several biocorrosion-relevant microbial groups (i.e., sulfate-reducing bacteria and acid-producing bacteria) was detected with commercially available test kits. Scanning electron microscopy and X-ray spectra analysis of osmium tetroxide-stained coupons demonstrated the development of microbial biofilm communities on some metal coupons submerged for 3 weeks in storage basins. After 12 months, coupons were fully covered by biofilms, with some deterioration of the coupon surface evident at the microscopical level. These results suggest that, despite the oligotrophic and radiological environment of the SRS storage basins and the active water deionization treatments commonly applied to prevent electrochemical corrosion in these facilities, these conditions do not prevent microbial colonization and survival. Such microbial densities and wide diversity of carbon source utilization reflect the ability of the microbial populations to adapt to these environments. The presumptive presence of sulfate-reducing bacteria and acid-producing bacteria and the development of biofilms on submerged coupons indicated that an environment for MIC of metal components in the storage basins may occur. However, to date, there has been no indication or evidence of MIC in the basins. Basin chemistry control and corrosion surveillance programs instituted several years ago have substantially abated all corrosion mechanisms.

Bacteriological Techniques↗
PubMed2001

A metabolic derivation of tritium transfer coefficients in animal products.

Tritium is a potentially important environmental contaminant originating from the nuclear industry, and its behaviour in the environment is controlled by that of hydrogen. Animal food products represent a potentially important source of tritium in the human diet and a number of transfer coefficient values for tritium transfer to a limited number of animal products are available. In this paper we present an approach for the derivation of tritium transfer coefficients which is based on the metabolism of hydrogen in animals. The derived transfer coefficients separately account for transfer to and from free (i.e. water) and organically bound tritium. A novel aspect of the approach is that tritium transfer can be predicted for any animal product for which the required metabolic input parameters are available. The predicted transfer coefficients are compared to available independent data. Agreement is good (R2=0.97) with the exception of the transfer coefficient for transfer from tritiated water to organically bound tritium in ruminants. This may be attributable to the particular characteristics of ruminant digestion. We show that tritium transfer coefficients will vary in response to the metabolic status of an animal (e.g. stage of lactation, diet digestibility etc.) and that the use of a single transfer coefficient from diet to animal product is inappropriate. It is possible to derive concentration ratio values from the estimated transfer coefficients which relate the concentration of tritiated water and organically bound tritium in an animal product to their respective concentrations in the animals diet. These concentration ratios are shown to be less subject to metabolic variation and may be more useful radioecological parameters than transfer coefficients. For tritiated water the concentration ratio shows little variation between animal products ranging from 0.59 to 0.82. In the case of organically bound tritium the concentration ratios vary between animal products from 0.15 (goat milk) to 0.67 (eggs).

Animals↗
PubMed2001

Fetal dose assessment for the offspring of the Techa Riverside residents.

Massive releases of fission products from the plutonium facility Mayak (Southern Urals, Russia) resulted in the contamination of the Techa river and its floodlands (1949-1956). The results of long-term studies of exposure populations have been used for different purposes of retrospective dosimetry. The riverside residents were exposed via various pathways and their progeny were exposed in utero both to external radiation and to internal radiation from radionuclides ingested by the mothers prior to conception and during pregnancy. Fetal doses due to 90Sr from the maternal skeleton were estimated and compared with doses from other pathways. Individual red bone marrow (RBM) doses to the late fetuses were calculated on the basis of 90Sr contents measured in the maternal skeleton and the 90Sr transfer coefficients (TC) to the fetal skeleton were determined on the basis of the Techa river data. Two values of TC were assumed depending on the mothers' maturation status in the period of maximum releases: TC=0.2 for group 1 (adulthood of mothers in 1950) and TC=0.02 for group 2 (mothers not yet in menarche age in 1950). Fetal doses in both groups that resulted from the different TC values varied by a factor of about 5-8. Furthermore, the fetal RBM doses due to 90Sr from the maternal skeleton were found to depend on the distances from the site of release and the time after major 90Sr intake. The average fetal RBM doses in the population of the upper, middle and lower Techa riverside regions were close to the ratio 3:2:1 and 20 years after the onset of contamination, the fetal doses have decreased by a factor of about 3-5. The distance from the site of release determined the relative contribution of different pathways to the fetal dose. For the settlements that are located closest to the site of release, the external exposure was of major concern up to the date of evacuation. For the non-evacuated settlements, the contribution of internal doses was higher and after 1956, external exposure is assumed to be negligible. From 1956 on, 90Sr that has been transferred from the maternal skeleton was practically the single source of in utero exposure.

Adult↗
PubMed2002

The Southern Urals radiation studies. A reappraisal of the current status.

In the late 1940s and early 1950s the nuclear workers of the Mayak Production Association in the Southern Urals were exposed to high doses from gamma-rays and from incorporated plutonium. In addition, the population of the Techa riverside downstream of the plutonium-production sites received continued exposures from external gamma-rays due to fission products released into the river and from the internal radiation due to incorporation of the fission products. Based on two international coordination meetings in 1998 and 2000, a synopsis has been given recently in this journal of the radioepidemiological studies on these exposed populations. This commentary describes the current status of these singular investigations with regard to the dosimetry, the assessment of late health effects, and the risk estimation both for the Mayak nuclear workers and the Techa riverside population. A central issue are newly published reduced estimates of the external dose to the Techa riverside population which imply substantially increased risk coefficients for solid cancer. Unless the new dosimetry system, TRDS-2000, has missed a major dose contribution, there is now conspicuous disagreement with current risk estimates. Unaccounted doses from atmospheric releases of fission products and from radiological screening of the Techa riverside population need to be explored, but underestimation of the short lived fission products released into the river appears to be a more critical factor. It is furthermore argued that even if TRDS-2000 were confirmed it would remain questionable whether risk estimates can be based on organ-specific doses when they are obtained in a population with a much higher bone-marrow exposure that may possibly have caused an 'abscopal' radiation effect.

Female↗
PubMed1995

Do persons living near a uranium processing site have evidence of increased somatic cell gene mutations? A first study.

The objective of this study was to examine if individuals living near a uranium processing site have greater mutagenic damage, as measured by three mutagenicity assays, compared with subjects unexposed to any nuclear facilities. The design was a cross-sectional exploratory analysis of 112 subjects; 56 volunteer residents were from within a 5-mile radius of the Fernald Uranium Processing site and 56 'control' subjects were from a geographically separate area unexposed to any known uranium emissions. The groups were constrained to be similar in age and sex composition. The main outcome measures were three human somatic gene mutation assays consisting of the HPRT T-lymphocyte cloning assay to measure 6-thioguanine resistant lymphocytes; the glycophorin A assay to detect the loss of expression of the M or N allele; and the micronucleus assay as a marker of chromosomal damage. The results showed no statistically significant or quantitatively important differences between groups for all three mutagenicity assays; only the unselected cloning efficiency was statistically significantly different between groups (0.42 +/- 0.16 for the Fernald versus 0.35 +/- 0.12 for the comparison groups). In both groups, age was significantly related to HPRT mutant frequency, with a 1.25% rate of increase in mutant frequencies for each 1-year gain of age in the Fernald group and a 1.12% rate of increase in mutant frequencies for each 1-year gain of age in the comparison group. For the micronucleus data, females had a greater mean micronucleus frequency than males. In addition, smokers had an increased mean ln (natural logarithm) HPRT mutant frequency (3.06 +/- 0.14 for current smokers compared with a mean of 2.72 +/- 0.05 for non-current (i.e. never plus former) smokers). Our results are consistent with the previously reported association between sex type and micronucleus frequency, the known relationship between age and T-lymphocyte cloning efficiency and age and HPRT mutant frequency, and verify the wide inter-subject variability for the latter. Finally, we conclude that at a population level, the relationships between current cigarette use and HPRT mutant frequency, and sex type and micronucleus frequency, are stronger than is the association between geographic proximity to a uranium processing site and mutagenic abnormalities.

Aged↗