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Biomedical subjects

T G Hinton

Publications and source records attributed to T G Hinton.

At least 19 recordsLinked to original sources

Radiation-induced untargeted germline mutations in Japanese medaka.

Radiation has been shown to increase mutation frequencies at tandem repeat loci by indirect interactions of radiation with DNA. We studied germline mutations in chronically exposed Japanese medaka (Oryzias latipes) using microsatellite loci. After screening 26 randomly selected loci among unirradiated parents and their 200 offspring, we selected seven highly mutable loci (0.5-1.0 x 10(-2) mutants per locus per gamete) and two bonus loci for further study. To determine if radiation exposure increases mutation frequencies in these loci, medaka were chronically irradiated from subadults through maturation at relatively low dose rates of 68 mGy/d. Total doses for males and females were 10.4 and 3 Gy, respectively. The mean number of mutations for the offspring of exposed families (0.149+/-0.044) was significantly higher (P=0.018) than for control families (0.080+/-0.028), indicating induction of germline mutations from chronic irradiation. This increase in the microsatellite mutation rate is greater than expected from direct interaction of radiation with DNA, suggesting indirect, untargeted mechanism(s) for mutations. This study identified microsatellite loci with a high mutational background in medaka, variation among loci and families as important variables, and demonstrated the usefulness of this fish model for studying radiation-induced germline mutations.

Alleles↗

Foliar uptake of cesium from the water column by aquatic macrophytes.

The probable occurrence and rate of foliar absorption of stable cesium (133Cs) from the water column by aquatic macrophyte species was analyzed following the addition of 133Cs into a small reservoir near Aiken, South Carolina, USA. An uptake parameter u (10(3)Lkg(-1)d(-1)) and a loss rate parameter k (d(-1)) were estimated for each species using time series of 133Cs concentrations in the water and plant tissues. Foliar uptake, as indicated by rapid increases in plant concentrations following the 133Cs addition, occurred in two floating-leaf species, Brasenia schreberi and Nymphaea odorata, and two submerged species, Myriophyllum spicatum and Utricularia inflata. These species had values of u> or =0.75 x 10(3)Lkg(-1)d(-1). Less evidence for foliar uptake was observed in three emergent species, including Typha latifolia. Ratios of u to k for B. schreberi, M. spicatum, N. odorata and U. inflata can be used to estimate concentration ratios (CR) at equilibrium, and these estimates were generally within a factor of 2 of the CR for 137Cs for these species in the same reservoir. This correspondence suggests that foliar uptake of Cs was the principal absorption mechanism for these species. Assessments of: (1) the prevalence of foliar uptake of potassium, rubidium and Cs isotopes by aquatic macrophytes and (2) the possible importance of foliar uptake of Cs in other lentic systems are made from a review of foliar uptake studies and estimation of comparable u and k values from lake studies involving Cs releases.

Cesium Radioisotopes↗

The influence of a whole-lake addition of stable cesium on the remobilization of aged 137Cs in a contaminated reservoir.

To document the short-term dynamics of Cs, 4 kg of (133)Cs were introduced into an 11.4-ha, 157 000 m(3) reservoir previously contaminated with (137)Cs from past reactor operations at the US Department of Energy's Savannah River Site near Aiken, South Carolina, USA. The (133)Cs addition resulted in an increase of 6.1 MBq of (137)Cs (1.9 mug (137)Cs) in the water column over the following 260 days. Possible sources for the increased (137)Cs included (1) release from the sediments, (2) release from the approximately 26 000 kg of aquatic macrophytes that occupied 80% of the reservoir, and (3) wash-in from the pond's watershed. Data are presented to indicate that release from the sediments was the principal source of the (137)Cs increase. The fraction of (137)Cs released from the sediments (0.7%) is consistent with laboratory measurements of (137)Cs desorption from neighboring ponds on the Savannah River Site.

Adsorption↗

Low Dose Rate Irradiation Facility: initial study on chronic exposures to medaka.

Uncertainties associated with the effects from chronic low-level exposures to radiation prompted us to construct a Low Dose Rate Irradiation Facility (LoDIF). The facility was designed specifically to test the appropriateness of the 10 mGy d(-1) guideline often espoused as acceptable for protection of aquatic biota from ionizing radiation. Scientists at the 0.4 ha facility use 40 outdoor mesocosms and 137Cs irradiators of three different source strengths to research the effects of chronic low-level irradiation at different levels of biological organization. A description of the facility is included along with results from a pilot study in which Japanese medaka (a small fish native to Asia) were chronically irradiated at the highest dose rate possible within the facility (350+/-150 mGy d(-1)). Irradiated fish produced fewer eggs per day (p=0.03); had a lower percentage of viable eggs (p=0.04), and produced a lower percentage of hatchlings (p=0.05). Although these data are not surprising based on the relatively high dose rates, they are important to future work at the LoDIF because they confirm the utility of our chosen model organism for detecting population-level responses, and they illustrate the statistical power achieved from using replicated mesocosms, in that statistical significance was achieved with few replicates per treatment. Future directions for the LoDIF are presented, as well as an invitation for interested researchers to participate in our studies.

Animals↗

The maximally exposed individual--comparison of maximum likelihood estimation of high quantiles to an extreme value estimate.

The dose to human and nonhuman individuals inflicted by anthropogenic radiation is an important issue in international and domestic policy. The current paradigm for nonhuman populations asserts that if the dose to the maximally exposed individuals in a population is below a certain criterion (e.g., <10 mGy d(-1)) then the population is adequately protected. Currently, there is no consensus in the regulatory community as to the best statistical approach. Statistics, currently considered, include the maximum likelihood estimator for the 95th percentile of the sample mean and the sample maximum. Recently, the investigators have proposed the use of the maximum likelihood estimate of a very high quantile as an estimate of dose to the maximally exposed individual. In this study, we compare all of the above-mentioned statistics to an estimate based on extreme value theory. To determine and compare the bias and variance of these statistics, we use Monte Carlo simulation techniques, in a procedure similar to a parametric bootstrap. Our results show that a statistic based on extreme value theory has the least bias of those considered here, but requires reliable estimates of the population size. We recommend establishing the criterion based on what would be considered acceptable if only a small percentage of the population exceeded the limit, and hence recommend using the maximum likelihood estimator of a high quantile in the case that reliable estimates of the population size are not available.

Animals↗

Effects of radiation on the environment: a need to question old paradigms and enhance collaboration among radiation biologists and radiation ecologists.

A historical perspective is given of the current paradigm that does not explicitly protect nonhuman biota from radiation but instead relies on the concept that if dose limits are set to protect humans, then the environment is automatically protected as well. We summarize recent international questioning of this paradigm and briefly present three frameworks for protecting biota that are being considered by the U.S. Department of Energy, the Canadian Nuclear Safety Commission, and the International Commission on Radiological Protection. We point out a controversial component in each of the three frameworks and suggest topics that need additional research. We emphasize that to properly address radiation protection of the environment, we need to understand how effects are integrated across different levels of biological organization. We caution that the proposed use of molecular end points to estimate ecological risks from radioactive contamination is applicable only if we understand the extent of the impact that molecular damage has on individual organisms and populations of exposed biota. To accomplish the latter, enhanced collaborations are required among the traditionally separate disciplines of radiation biology and radiation ecology.

Conservation of Natural Resources↗

Reduction of crop contamination by soil resuspension within the 30-km zone of the Chernobyl nuclear power plant.

A field experiment was conducted within the 30-km zone of the Chernobyl Nuclear Power Plant to analyze whether the application of mulching reduced resuspension of 137Cs contaminated soil in oat (Avena sativa) crops. In 1993, we applied a mulch treatment at a dose of 200 g m(-2), and soil resuspension was measured by estimating soil loadings onto plant surfaces from Ti concentrations in plants. In 1994, two mulch doses were applied, 200 and 50 g m(-2), and we estimated the contribution of soil resuspension by using artificial resuspension collection devices (ARC). In the 1993 experiment between 4.6 and 34.4% of the plant's total 137Cs contamination was attributed to external soil contamination. The mean amount of soil-derived 137Cs attached to vegetation was 124.7 Bq kg(-1)(plant) in control plots and 53.7 Bq kg(-1)(plant) in mulched plots. In the 1994 experiment, covering the soil with a mulch layer decreased the radiocesium content in ARC by about 70%. Results obtained in these experiments suggest that soil resuspension was a significant mechanism for plant contamination and that mulching was effective in reducing that contamination.

Agriculture↗

Environmental biodosimetry: a biologically relevant tool for ecological risk assessment and biomonitoring.

Biodosimetry, the estimation of received doses by determining the frequency of radiation-induced chromosome aberrations, is widely applied in humans acutely exposed as a result of accidents or for clinical purposes, but biodosimetric techniques have not been utilized in organisms chronically exposed to radionuclides in contaminated environments. The application of biodosimetry to environmental exposure scenarios could greatly improve the accuracy, and reduce the uncertainties, of ecological risk assessments and biomonitoring studies, because no assumptions are required regarding external exposure rates and the movement of organisms into and out of contaminated areas. Furthermore, unlike residue analyses of environmental media, environmental biodosimetry provides a genetically relevant biomarker of cumulative lifetime exposure. Symmetrical chromosome translocations can impact reproductive success, and could therefore prove to be ecologically relevant as well. We describe our experience in studying aberrations in the yellow-bellied slider turtle as an example of environmental biodosimetry.

Animals↗

Comparative bias associated with various estimates of dose to maximally exposed individuals.

Protection of the environment from anthropogenic radiation is a resurging international interest. The paradigm currently in use argues that the population is adequately protected if dose rates to the maximally exposed individuals are below a certain limit. Based on data sampled from natural populations, resource managers need to be able to test the hypothesis that dose rates to the maximally exposed individuals are acceptable. Recognizing the difficulty of sampling the maximally exposed individual within a contaminated environment, risk assessors have used various alternative approaches that vary from changing the paradigm and applying recommended dose rate limits to representatively, rather than maximally, exposed individuals, to using the 95th percentile of the sample mean as an estimator of the population maximum. To determine the effectiveness of numerous proposed alternatives, we used computer simulation techniques to generate a "population" of doses with known distributional qualities and then mathematically "sampled" the population to compare the ability of the various statistics at estimating the known population maximum. The simulation procedure was repeated 1000 times using Monte Carlo techniques, each time producing a measure of the distance between the estimate and the true value. We were thus able to quantify the bias associated with several approaches used to determine compliance with dose rate criteria established by the Department of Energy for protecting biota. The 95th quantile of the sample mean, and the sample maximum underestimated the population maximum by as much as 72 and 44%, respectively. The maximum likelihood estimate (MLE) of the 99.99th percentile was found to be the best predictor of the population maximum, even for small sample sizes of 20 and for both normally and lognormally distributed populations. However, bias associated with the MLE increased significantly if the population's distribution was incorrectly identified. We suggest shifting the regulatory criterion appropriately to argue that if the top 1% (as opposed to the maximum) of the population has a dose rate less than or equal to the regulatory limit then the population is adequately protected, and then using the MLE of the 99th percentile as the least biased sample statistic. Results of this study are also relevant when estimating dose to critical sub-groups of humans whose lifestyles are such that their doses are among the maximum for the population.

Animals↗

A comparison of 90Sr and 137Cs uptake in plants via three pathways at two Chernobyl-contaminated sites.

Foliar absorption of resuspended 90Sr, root uptake and contamination adhering to leaf surfaces (i.e. soil loading) were compared at two Chernobyl-contaminated sites, Chistogalovka and Polesskoye. Although foliar absorption of resuspended 90Sr was quantifiable, its contribution amounted to less than 10% of the plants' total, above-ground contamination. Root uptake was 200 times greater than foliar absorption at the near-field site of Chistogalovka and eight times greater at Polesskoye, where the fallout consisted of the more soluble condensation-type, rather than fuel particles. Strontium's bioavailability exceeded that of 137Cs (analyzed in the same plants) by orders of magnitude when compared using concentration ratios. Simplistic, cumulative effective dose calculations for humans ingesting 90Sr- and 137Cs-contaminated plants revealed that the dose at Chistogalovka was greater from 90Sr (185 mSv vs. 3 mSv from 137Cs), while at Polesskoye the dose from 137Cs (66 mSv) was 30 times greater than from 90Sr (2 mSv).

Absorption↗

Conservation of chromosome 1 in turtles over 66 million years.

Fluorescence in situ hybridization of a whole chromosome 1-specific probe from the yellow-bellied slider turtle (Trachemys scripta) to cells from four other species of turtle ranging from a desert tortoise to a loggerhead sea turtle resulted in specific and exclusive hybridization to chromosome 1 in all five species. Previous observations of conservation in the giemsa banding pattern and chromosome morphology and number among turtles are thus extended to the DNA sequence level, revealing a cytogenetic stability of chromosome 1 in these turtles during the past 66-144 million years. This contrasts with the situation for various hominoid species where, in many instances, extensive chromosomal rearrangements have been reported in one third of that time period. Our probe, which was prepared by microdissecting whole chromosomes from embryonic T. scripta fibroblasts and amplifying using DOP-PCR, is the first report of a whole-chromosome FISH probe for any reptile.

Animals↗

Chromosome translocations in T. scripta: the dose-rate effect and in vivo lymphocyte radiation response.

Using a whole-chromosome FISH painting probe we previously developed for chromosome 1 of the yellow-bellied slider turtle (Trachemys scripta), we investigated the dose-rate effect for radiation-induced symmetrical translocations in T. scripta fibroblasts and lymphocytes. The dose rate below which no reduction in effect per unit dose is observed with further dose protraction was approximately 23 cGy h(-1). We estimated the whole-genome spontaneous background level of complete, apparently simple symmetrical translocations in T. scripta lymphocytes to be approximately 1.20 x 10(-3)/cell projected from aberrations occurring in chromosome 1. Similar spontaneous background levels reported for humans are some 6- to 25-fold higher, ranging from about 6 x 10(-3) to 3.4 x 10(-2) per cell. This relatively low background level for turtles would be a significant advantage for resolution of effects at low doses and dose rates. We also chronically irradiated turtles over a range of doses from 0-8 Gy delivered at approximately 5.5 cGy h(-1) and constructed a lymphocyte dose-response curve for complete, apparently simple symmetrical translocations suitable for use with animals chronically exposed to radiation in contaminated environments. The best-fitting calibration curve (not constrained through the zero dose estimate) was of the form Y(as) = c + aD + bD(2), where Y(as) was the number of apparently simple symmetrical translocations per cell, D was the dose (Gy), a = (0.0058 +/- 0.0009), b = (-0.00033 +/- 0.00011), and c = (0.0015 +/- 0.0013). With additional whole-chromosome probes to improve sensitivity, environmental biodosimetry using stable chromosome translocations could provide a practical and genetically relevant measurement end point for ecological risk assessments and biomonitoring programs.

Animals↗

Resource allocation-based life histories: a conceptual basis for studies of ecological toxicology.

Whereas ecological assessments of contaminants are concerned with populations and higher levels of organization, most mechanistic work in toxicology is directed at effects on individuals and their parts. We propose that studies based on individuals can be useful in ecological analysis of polluted systems when based on the concepts of resource allocation-based life history analysis. At the heart of the resource allocation approach is the concept of operative environments of individuals (i.e., environmental factors influencing birth, death, or migration). Contaminants can have strong influences on operative environments, modifying resource allocation strategies that reflect changes in energy assimilation and demands. By examining contaminant-induced responses of individuals from the perspective of changing operative environments, individual-based changes and population dynamics can be addressed in an ecologically rigorous manner.

Animals↗

Culture methods for turtle lymphocytes.

Optimization of culture techniques for turtle and other reptilian lymphocytes is essential for facilitating cytogenetic and immunologic research for these animals. We examined a variety of conditions and parameters relevant to turtle lymphocyte culture including: different mitogenic agents, alone and in combination; lymphocyte separation protocols; culture volume; time required to stimulate lymphocytes to mitosis; importance of humidity and gas exchange in culture incubation; suitability of different culture media; effects of varying serum concentrations; ability of interleukin-2 (IL-2) to stimulate lymphocyte growth and prevent apoptosis; and feasibility of inducing premature chromosome condensation. The best conditions of those we studied for obtaining mitotic cells were (1) the combined use of phytohemagglutinin-M form (2%) and lipopolysaccharides (0.55 microg/ml), (2) the use of 5% autologous turtle serum (as opposed to fetal bovine serum), and (3) collection of mitotic cells around 96 hours after mitogenic stimulation. Human, recombinant IL-2 did not increase the fraction of lymphocytes in mitosis over the range of concentrations tested and calyculin A was ineffective at inducing premature chromosome condensation in turtle lymphocytes over the range of concentrations tested. This test regime provides a guideline for determination of appropriate lymphocyte culture conditions in turtles and other reptiles.

Animals↗

Chromosome translocations in turtles: a biomarker in a sentinel animal for ecological dosimetry.

Nonhuman organisms are being exposed to ionizing radiations at radionuclide-contaminated sites around the world. Direct methods are seldom available for measuring biologically relevant doses received by these organisms. Here we extend biological dosimetry techniques, which are much better developed for humans and a few other mammalian species, to a nonmammalian species. Turtles were chosen because a long-lived animal would best serve the need for low-level, chronic exposure conditions. We chose the yellow-bellied slider turtle (Trachemys scripta), which is known to have a maximum life span of at least 22 years. As reported elsewhere, we first isolated an embryonic fibroblast cell line and constructed whole-chromosome-specific DNA libraries for chromosome 1 by microdissection and PCR. A FISH painting probe was prepared and used to establish a dose-response curve for ionizing radiation-induced chromosome interchange aberrations in turtle fibroblasts. This was compared to the dose response for human fibroblasts treated under similar conditions in our laboratory. With respect to induction of chromosome interchange aberrations, human fibroblasts were approximately 1.7 times more sensitive than the T. scripta fibroblasts. To the extent that symmetrical interchanges are persistent over long periods, this approach could eventually provide a measure of the integrated lifetime dose these organisms receive from radionuclides in their environment and give a measure of the extent of relevant genetic damage over that time.

Animals↗

A comparison of techniques used to estimate the amount of resuspended soil on plant surfaces.

The objectives of this study were to compare four common techniques used to estimate soil mass loadings on plant surfaces and to assess the need to account for particle-size distributions of both the soil tracer and contaminant of concern within the soil. Soil loadings (g soil kg-1 dried plant) from split samples collected in a pasture near Chernobyl were estimated using soil tracers of plutonium analyzed via alpha spectroscopy (mean +/- standard error; 1.0 +/- 0.2), titanium analyzed with an inductive coupled plasma spectrometer; (3.6 +/- 0.6), and neutron activation analysis for scandium (8.1 +/- 1.6), as well as simply washing the soil off the vegetation (34.1 +/- 5.6) Differences were significant at p < 0.001. We also found that soil loading estimates from any one technique varied by a factor of 10 depending on the soil particle size used in the calculations. This was because soil loadings decreased when smaller-sized soil fractions dominated the resuspension process. However, the percent of the plant's total contamination attributable to soil loading increased with smaller soil particles. Smaller soil particles apparently contribute less to the mass of soil loading (g soil kg-1 dry plant), but more to the total plant contamination (Bq) because of the higher concentration of contaminant found in the smaller-sized soil fractions. Differences in mass loading estimates due to the technique chosen (a factor of 10), or due to differences in elemental concentration as a result of the soil particle size used in the calculation (also a factor of 10), were greater than the natural variability observed in the field (2.5).

Alpha Particles↗

Sensitivity analysis of ECOSYS-87: an emphasis on the ingestion pathway as a function of radionuclide and type of deposition.

A sensitivity analysis of parameters associated with the ingestion pathway was conducted for the computer model ECOSYS-87. The model is currently being used extensively throughout Europe for accident consequence analysis following a nuclear release. Individual parameter perturbation was used to develop sensitivity indices. The sensitivity indices identified parameters whose uncertainties had a large impact on model results. The relative ranking of the sensitive parameters depended on the radionuclide (137Cs, 90Sr, 131I, or 239Pu), whether dose 1 y or 50 y postaccident was being considered, and whether the deposition event was dry or mixed. The most influential parameter for 239Pu was resuspension. Parameters to which human dose was sensitive following exposure to 137Cs, 90Sr, and 131I, were as follows: yield (biomass) of vegetation, transfer of radionuclides from plants to animals, deposition velocity, changes in radionuclide concentrations due to food processing, livestock feeding rates, and weathering of radionuclides from plant surfaces. The ranking of 131I's parameters was governed by its 8-d physical half-life. Parameters that affected the initial deposition, parameters that could rapidly affect the transfer of 131I from the biota to humans, or parameters that allowed 131I to decay prior to consumption were important. Important parameters specific to 90Sr and 137Cs included transfer of radionuclides from soil to plant, leaching from the plant rooting zone, and resuspension. Parameters associated with the movement of radionuclides within the soil were not as important for 137Cs as they were for 90Sr. With the exception of the deposition velocity, if a parameter proved to be sensitive for dry deposition, it was as sensitive, or even more so, for a mixed deposition event. Extending the model end point from 1 y to 50 y postaccident also caused a shift in the relative ranking of sensitive parameters for 137Cs and 90Sr. Parameters that were not important for any of the radionuclides considered under this scenario were those related to timing and length of crop harvest, transfer of radionuclides from leaf surfaces to edible portions of plants, rate at which radionuclide concentrations in plants decrease due to growth dilution, and time for animals to reach marketable size.

Accidents↗