Cataract in the chronically exposed residents of the Techa riverside villages

The present study is based on a retrospective analysis of archive data of the Clinical Department of the Urals Research Center for Radiation Medicine that has been established to examine and treat accidentally exposed residents of the Urals Region. All individuals included in this study were examined by an ophthalmologist. The study of cataract incidence has been conducted retrospectively for the period from 1951 till 2000 among chronically exposed residents of the Techa riverside villages (6343 persons). Individual accumulated absorbed doses to soft tissues (analogue of eye dose) reached 1.18 Gy (mean 0.12 Gy) while for 88.9% of the study group the dose did not exceed 0.1 Gy. There was no evidence of the influence of low-dose and low-dose rate on cataract incidence. Excess relative risk of cataract formation per 1 Gy was 0.40 (95% CI ?0.43; 1.47). It is noted that 15% of all excess cases were registered in persons with soft tissue dose above 0.3 Gy, though their fraction among all examined persons was only 4.1%. Risk of cataract development significantly increased in exposed individuals with retinal angiosclerosis, diabetes and arterial hypertension.     

Modeling analysis of the lymphocytopoiesis dynamics in chronically irradiated residents of Techa riverside villages

A biologically motivated dynamical model of the lymphocytopoietic system in irradiated humans is applied here to analyze the data obtained under hematological examinations of residents of Techa riverside villages. Those people were exposed to chronic irradiation with varying dose rates, due to the radioactive contamination of the river basin by the Mayak Production Association. Modeling studies revealed the relationship between the dynamics of the lymphocytopoietic system in the examined individuals and the variation of dose rate over the considered period of time. It is found that the developed model is capable of reproducing the decreased level of blood lymphocyte concentration observed during the period of maximum radiation exposure, the recovery processes in the system observed during the period of decreasing dose rate, as well as the enhanced mitotic activity of bone marrow precursor cells in this hematopoietic lineage observed during the entire period under consideration. Mechanisms of these effects of chronic irradiation on the human lymphocytopoietic system are elucidated based on the applied model. The results obtained demonstrate the efficiency of the developed model in the analysis, investigation, and prediction of effects of chronic irradiation with varying dose rate on the human lymphocytopoietic system. In particular, the developed model can be used for predicting any radiation injury of this vital system in people exposed to chronic irradiation due to environmental radiological events, such as anthropogenic radiation accidents or radiological terroristic attacks.     

Characterization of immunity status in exposed residents of the Techa riverside villages 50 years after the onset of radiation exposure

The goal of the study was to assess the state of immunity in exposed residents of the Techa riverside villages 50 years, or more, after the onset of radiation exposure. 127 chronically exposed persons and 55 unexposed persons were studied. The mean dose to red bone marrow (RBM) was 0.69 Sv in exposed subjects, the mean dose to soft tissue was 0.07 Sv, the mean dose rate amounted to 0.10 Sv/yr to RBM and 0.02 Sv/yr to soft tissues in 1950. The state of the basic links of the immunity system (cellular, humoral, mononuclear phagocyte system, cytokine spectrum, etc.) was assessed using conventional methods. Exposed persons manifested a significant reduction in the absolute counts of CD3+, CD4+, CD 11b+, CD16+ lymphocytes in the peripheral blood, as well as an increase in the relative counts of CD8+. The group comprised of the Techa riverside residents demonstrated an increased immunoregulatory index (exposed individuals: 1.47; controls: 1.71, p = 0.001). An increased production of Immunoglobulin A and increased proportions of CD25+ lymphocytes were revealed in exposed individuals. Changes in the phagocytic activity of neutrophils and monocytes were insignificant, and were primarily associated with changes in the proportions of pagocytes in the peripheral blood stream. The state of the immunity in chronically exposed individuals at late time after the begin of exposure is characterized by a number of specific features reflected primarily on the cellular immunity. No relationship between immunity changes and accumulated exposure dose and dose rate were noted over the period of maximum radiation exposures (1950).     

Status of hemapoiesis in residents of the Techa riverside villages in the period of maximum radiation exposure. Report 2. Influence of exposure dose and dose rate of red bone marrow as well as modifying factors on the frequency of cytopenia and cytosis

The purpose of this study is a retrospective estimation of the influence of dose and dose rate of the red bone marrow chronic radiation exposure in combination with various modifying factors (gender, age, comorbidity) on the frequency of deviations from normal values of the results of peripheral blood investigation in humans exposed on the Techa River. The results of investigation show that humans chronically exposed to radiation can develop marked changes in the cellular composition of peripheral blood characterized by a tendency to cytopenia (signs of the decompensation of hemopoiesis). The tendency to cytopenia can be identified earlier in the lymphoid germ, and later in platelet and erythroid lines. A high lability of granulocytes under the influence of various, often infectious, factors is the cause of the lack of statistically significant differences in terms of frequency of neutropenia. Several non-radiation factors (gender, age, health status) in combination with radiation exposure could have a modifying influence on hematopoiesis, which contributed to the disruption of adaptation processes and the development of conditions characterized by a tendency to cytopenias in exposed individuals. The red bone marrow dose rate reduction resulted in a gradual decrease in the frequency of erythrocytopenia, thrombocytopenia, neutropenia and lymphocytopenia in the group of exposed population. Increased frequencies of erythrocytosis, thrombocytosis, lymphocytosis, monocytosis and neutrophilia were observed when the median dose rate was reduced to the level of 0.024 Gy/year (in the year 1956), which could be regarded as activation of regenerative processes in hematopoiesis.     

Radioecological situation at the villages of the Techa river

The Techa river was contaminated in 1949-1956 from the nuclear enterprise "Mayak". The investigations were carried out in flood plain of the Techa river in 1992-2001. 90Sr and 137Cs stores were calculated in the soil-vegetation cover. There is uneven character of the spatial radionuclides contamination of the investigated area. The store with 90Sr changes from 25 to 930 kBq/m2 (0.7-25.0 Ci/km2) and that with 137Cs--from 30 to 1700 kBq/m2 (0.8-46.0 Ci/km2). In the preriver-bed soils the ratio 90Sr/137Cs increases with further from discharge point. Individual effective dose was calculated for the Brodocalmak population. 90Sr was revealed in the flood plain soils of the Iset river. The contribution of the contaminated Techa river and its flood plains soils accounted for as by incorporated radionuclides as background gamma-radiation does not exceed 0.13-0.17 mSv/yr if the contaminated Techa river utilization is limited. In other case the contribution of the contaminated Techa river increases to 1.6-3.0 mSv/yr. These values exceed international safety norms.     

In vitro hemopoiesis in human micro long-term bone marrow cultures recharged with either allogeneic, T-cell-depleted allogeneic, or syngeneic bone marrow cells

The production of granulocyte-macrophage colony-forming cells (GM-CFC) and the proliferation period in human long-term bone marrow cultures are inferior to murine cultures. There is also evidence that recharge of the cultures after establishing confluent stromal layers will not greatly improve myelopoiesis. Data in the literature indicate that PHA-responsive T lymphocytes persist for up to 5 weeks in human but not in murine long-term marrow cultures. We therefore analyzed the effects of recharging micro long-term bone marrow cultures with bone marrow cell samples depleted by T lymphocytes. Depletion was performed in a complement-mediated cytotoxicity assay by applying the monoclonal antibody CAMPATH-1. Our data show that regardless of whether T cells were removed only at recharge, at both initiation and recharge, or only at initiation, obvious enhancement could neither be achieved in the GM-CFC production nor in the proliferation period. Furthermore, no advantage was seen when using syngeneic marrow cells. We conclude that in allogeneic long-term marrow cultures hemopoiesis is not limited by immunological incompatibilities.     

Reconstruction of 90Sr intake for breast-fed infants in the Techa riverside settlements

The Techa River (Southern Urals, Russia) was contaminated as a result of radioactive releases by the Mayak plutonium production facility during 1949-1956. The persons born after the onset of the contamination have been identified as the "Techa River Offspring Cohort" (TROC). The TROC has the potential to provide direct data on health effects in progeny that resulted from exposure of a general parent population to chronic radiation. The purpose of the present investigation is the estimation of (90)Sr intake from breast milk and river water in the period from birth to 6 months of life, necessary for an infant dose calculation. The investigation is based on all available data concerning radioactive contamination due to global fallouts and Mayak releases in the Southern Urals where extensive radiometric and radiochemical investigations of human tissues and environmental samples were conducted during the second half of the twentieth century. The strontium transfer factor from mother's daily diet to breast milk was estimated as 0.05 (0.01-0.13) d L(-1). Based on this transfer factor and data on (90)Sr water contamination, the average total (90)Sr intake for an infant born in the middle Techa River region was found to be equal to 60-80 kBq in 1950-1951. For the same period, calculations of (90)Sr intake using ICRP models gave values of 70-100 kBq. From 1952 onwards, the differences in intakes calculated using the two approaches increased, reaching a factor of 2-3 in 1953. The Techa River data provide the basis for improving and adapting the ICRP models for application to Techa River-specific population.     

The xid mutation affects hemopoiesis in long term cultures of murine bone marrow

Cells of the humoral immune system are particularly affected by a mutation at the X chromosome linked immunodeficiency disease (xid)locus. Although B cells are made in normal numbers, they fail to become phenotypically and functionally diverse. Consequently, poor antibody responses are mounted to certain types of Ag. There have been some indications that other types of hemopoietic cells may be influenced by the mutation and development of the humoral immune system is unusually dependent on the presence of T lymphocytes. We now describe an analysis of the lympho-hemopoietic environment studied with long term bone marrow cultures. Contrary to expectations, cultures initiated with cells from homozygous female or hemizygous male mice with the mutant allele established more quickly than normal. The accelerated initial growth pattern was clearly linked to the xid mutation. Artificial mixtures of marrow exhibited intermediate growth kinetics. Experiments with H-2 congenic and T6 chromosome marked cells did not reveal an intrinsic dominance of growth in nonadherent xid cells. Similar results were obtained with culture conditions which favored production of myeloid or lymphoid cells. These findings would be consistent with subtle changes in the bone marrow microenvironment resulting from the xid mutation. The pedigree of the mouse strains had a significant influence on lymphopoiesis in long term bone marrow cultures. Lymphocytes of BALB/c origin dominated over CBA/H background cells in cultures established from mixtures of the two, but this did not correlate with any functional deficiency in CBA/H stromal cells. In fact, establishment of an adherent layer was a rate-limiting step in initiating long term cultures and this could be achieved with a low dose inoculum of CBA/H marrow. Even more dramatic effects were found in hemopoietic cells from doubly defective C3H.nu/nu-xid mice. The bone marrow of these athymic animals contained normal numbers of granulocyte/macrophage progenitors. However, lymphoid cultures could not be reproducibly established with their cells and myelopoiesis was never observed in vitro. The relatively simple conditions which pertain in culture make it possible to appreciate effects of mutations and pedigree on hemopoiesis which are unremarkable in intact animals.     

Status of hemapoiesis in residents of the Techa riverside villages in the period of maximum radiation exposure. Report 1. Evaluation of the cellular composition of peripheral blood and the role of comorbidities in the oppression of hemapoiesis

Residents of the Techa riverside villages chronically exposed to ionizing radiation (the average dose rate in 1951-1956 was 0.047 Gy/year, the maximum reached 2.44 Gy/year) developed marked changes in the cellular composition of peripheral blood. The maximum reduction of peripheral blood counts occurred in the years 1951-1953, after which the beginning of the restoration of cellularity to the control level was observed. The dose rate at this point (1956) was about 0.02 Gy/year. The factors of radiation and non-radiation nature (gender, the age at the onset of exposure, health related disorders) in different combinations affect the number of peripheral blood cells. The influence of dose rate of a chronic radiation exposure on a platelet count takes precedence over other factors. The factors of gender and dose rate determine the number of erythrocytes in exposed persons. The changes accompanying the health status and dose rate significantly affect the number of neutrophils. The influence of comorbidity, age and dose rate on the number of monocytes was noted. A lymphocyte count was mainly determined by the age at the onset of exposure and concomitant diseases. A joint influence of chronic radiation exposure and concomitant diseases increases a mutual action on erythro- and thrombocytopoiesis. The decrease of the dose rate was followed by a gradual predominance of the somatic disease influence on leukocyte (neutrophils, lymphocytes and monocytes) counts.     

Effect of indometofen on the survival and bone marrow hemopoiesis of mice exposed to acute external gamma- or X-ray irradiation

The purpose of the study is evaluation of radioprotective effectiveness of indometofen at its prophylactic administration in conditions of acute irradiation. Evaluation of radioprotective efficiency was performed by studying the 30-day survival rate, life expectancy, structure of deathly irradiated mice, and bone marrow hemopoiesis using methods of endogenous and exogenous colony formation. The prophylactic application of indometofen at doses 30 mg/kg for 5 days before irradiation has been observed to protect mice against radiation death induced by gamma or X-ray exposures at doses LD(50-70/30), increasing their survival rate by 16-44%, and reduce severity of post radiation disorders of bone marrow hemopoiesis.     

Cytogenetic study of the bone marrow and peripheral blood lymphocytes in monkeys in long-term gamma irradiation

A comparative study of chromosome structures of bone marrow and peripheral blood cells has been carried out in 8 rhesus macaques (2-7 years of age), 4 of which survived after prolonged low-capacity (3.87 microA/kg) gamma irradiation, the total dose being 7.97 Gy (LD50/60). It has been established that prolonged low-capacity gamma irradiation was of a high mutagenic activity. Various tissues of irradiated monkeys showed differences in the frequency (4 months) and types (4-33 months) of aberrations within the period of 4 to 33 months following irradiation. Mutagenic effect characteristic of the early period after the irradiation was retained in the peripheral blood of irradiated monkeys within the period of observation.     

Role of stromal microenvironment in the regulation of bone marrow hemopoiesis after curantyl administration

Role of the stromal microenvironment in regulation of bone marrow hemopoiesis at the administration of the thrombocyte disaggregant curantyl was studied by the method of heterotopic transplantation of the mice bone marrow. It is shown that the action of curantyl on hemopoiesis is realised through the stem stromal cells of the bone marrow. It is noted that the inhibitory action of the preparation on proliferation of osteogenic precursor-cells is followed by activation of bone resorption processes in regenerating ectopic hemopoietic organ. Under the action of curantyl at low bone marrow cellularity in the focus of heterotopic hemopoiesis and femur an increase of mitotic activity in hemopoietic elements is noted. It is revealed that a phenomenon of ineffective megakaryocytopoiesis with intramedullary destruction of megakaryocytes leads to the local excretion of the thrombocyte released growth factor (TRGF) which has a compensatory character.     

Proliferation and differentiation of bone marrow stromal cells under hypoxic conditions

Low oxygen tension is a potent differentiation inducer of numerous cell types and an effective stimulus of many gene expressions. Here, we described that under 8% O(2), bone marrow stromal cells (MSCs) exhibited proliferative and morphologic changes. The level of differentiated antigen H-2Dd and the number of G(2)/S/M phase cells increased evidently under 8% O(2) condition. Also, the proportion of wide, flattened, and epithelial-like cells (which were alkaline phosphatase staining positive) in MSCs increased significantly. When cultured in adipogenic medium, there was a 5- to 6-fold increase in the number of lipid droplets under hypoxic conditions compared with that in normoxic culture. We also demonstrated the existence of MSC differentiation under hypoxic conditions by electron microscopy. Expression of Oct4 was inhibited under 8% O(2) condition, but after adipocyte differentiation in normoxic culture and hypoxia-mimicking agents cobalt chloride (CoCl(2)) and deferoxamine mesylate (DFX) treatments, Oct4 was still expressed in MSCs. These results indicate hypoxia accelerates MSC differentiation and hypoxia and hypoxia-mimicking agents exert different effects on MSC differentiation.     

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. Received: 8 December 2000 / Accepted: 1 July 2001