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Introduction: Epidemiological studies indicate that obesity is associated with an increased risk of 20-25% with several types of cancer., Objective: The frequency of chromosome aberrations was evaluated in lymphocytes from postmenopausal obese and non-obese women., Materials and Methods: Twenty obese and 20 non-obese women, all post-menopause, were recruited. The groups were matched according to age (± 5 years) and place of origin. After signing the consent form, women were interviewed using a structured questionnaire, and a blood sample (5 ml) was drawn into vacutainer tubes. From each sample, lymphocyte cell cultures were established with and without mitomycin C (challenge assay). Afterwards, the frequency of chromosome aberrations were recorded for each group and treatment. Data were analyzed using the statistical program SPSS, v. 14.0., Results: Obese women had a higher frequency of chromosome aberrations when compared with non-obese women. After exposing the cell cultures to mitomycin C, obese women presented an increase in the number of total chromosome aberrations in comparison to non-obese women (3.7± 0.6 vs. 2.70±0.6; p=0.001)., Conclusions: The higher frequency of chromosome aberrations in lymphocytes from postmenopausal obese women compared to non-obese women suggested differences in the DNA repair capacity. This may indicate an association between genomic instability and the higher incidence of cancer in this population.

Introduction: Mutagens contained in complex mixtures can present synergistic interactions, either additive or antagonistic. Therefore, development of experimental approaches is necessary to elucidate which is the responsible agent for the effect in the mixtures., Objective: An experimental design was developed that allowed an understanding of the processes between the compounds of complex mixtures., Materials and Methods: Human lymphocytes were exposed to binary mixtures of the mutagens B[a]P, DMBA, Trp-P-1 and MX for 1 hour with or without S9. Viability was assessed with trypan blue dye and the genotoxicity by the comet assay., Results: All of the hydrocarbon showed an effect with furanone. With and without S9, the most toxic interactions were observed between hydrocarbons. Synergistic interaction was observed without S9 between B [a] P and Trp-P-1 and between DMBA and Trp-P-1 with metabolic activity. Without S9 antagonistic interaction was observed only between Trp-P-1+DMBA, and with S9 between Trp-P-1+MX and MX+DMBA. It observed an increase dose dependent in tail length. Half the cultures showed genotoxic damage and increased cell damage. For each mixture, minimum concentrations were determined at which adverse effects are observed; for some only the maximum concentration was determined at which no adverse effects are observed., Conclusion: The processes between mutagens present in a mixture have become better understood, and the results validated an analytical model that determined which component had an effect on another. The results also showed that the type of compounds in the mixture determined whether or not a risk threshold was present.

The recent steep increase in population dose from radiation-based medical diagnostics, such as computed tomography (CT) scans, requires insight into human health risks, especially in terms of cancer development. Since the induction of genetic damage is considered a prominent cause underlying the carcinogenic potential of ionizing radiation, we quantified the induction of micronuclei and loss of heterozygosity events in human cells after exposure to clinically relevant low doses of X rays. A linear dose-response relationship for induction of micronuclei was observed in human fibroblasts with significantly increased frequencies at doses as low as 20 mGy. Strikingly, cells exposed during S-phase displayed the highest induction, whereas non S-phase cells showed no significant induction below 100 mGy. Similarly, the induction of loss of heterozygosity in human lymphoblastoid cells quantified at HLA loci, was linear with dose and reached significance at 50 mGy. Together the findings favor a linear-no-threshold model for genetic damage induced by acute exposure to ionizing radiation. We speculate that the higher radiosensitivity of S-phase cells might relate to the excessive cancer risk observed in highly proliferative tissues in radiation exposed organisms.

We describe a low level of chromatid-type aberrations which included the relatively rare isochromatid/chromatid triradial in peripheral blood lymphocytes that were irradiated, ostensibly in G0, with accelerated heavy (12)C ions. These were produced only at the energies of 69 MeV/n (34.6 keV/microm), almost absent at the energy of either 58.6 MeV/n (46.07 keV/ microm) or 19.3 MeV/n (97 keV/microm), nor were they found after low-LET X-rays. Mechanisms potentially responsible for their formation are discussed.

Biological cells are complex in both morphological and biochemical structure. The effects of cellular fine structure on light scattered from cells are studied by employing a three-dimensional code named AETHER which solves the full set of Maxwell equations by using the finite-difference time-domain method. It is shown that changes in cellular fine structure can cause significant changes in the scattered light pattern over particular scattering angles. These changes potentially provide the possibility for distinguishability of cellular intrastructures. The effects that features of different intrastructure have on scattered light are discussed from the viewpoint of diagnosing cellular fine structure. Finally, we discuss scattered light patterns for lymphocyte-like cells and basophil-like cells.

Published data concerning the effects of indoor radon exposure on the frequency of chromosome aberrations in peripheral lymphocytes of residents are contradictory. Possible reasons for this may be the low radon concentration in dwellings and/or the limited number of investigated persons. We therefore studied the relationship of domestic radon exposure and the occurrence of chromosome aberrations in peripheral lymphocytes in 61 persons living in houses with radon concentrations from 80 up to 13,000 Bq/m3. We analyzed 60,000 cells from fluorescence plus Giemsa (FPG)-stained slides. It could be clearly demonstrated that in groups of persons living in dwellings with indoor radon concentrations >200 Bq/m3 the number of cells containing dicentrics and/or centric rings (C(dic + cr)) (2.45 +/- 0.50 x 10(-3)) was significantly increased (p < 0.05) in comparison to the control level (1.03 +/- 0.15 x 10(-3)). However, there was no difference in the mean frequency of C(dic + cr) between the groups living in dwellings with higher radon concentrations. Using the fluorescence in situ hybridization (FISH) technique for the detection of translocations, we analyzed 23,315 cells in 16 persons of the highest exposed group (>5,000 Bq/m3). The observed frequency of translocations was 3.9 +/- 0.64 x 10(-3). In comparison to the control group (2.02 +/- 0.18 x 10(-3)), there was a slight but not statistically significant increase in the exposed group (P = 0.055). If, however, the age of the examined persons is taken into account, the values are significantly increased (P < 0.05) in the exposed persons older than 40 years in comparison to the age-matched controls. Since most of the translocations were found in stable cells, it is concluded that translocations are also induced in blood-forming tissue and are transmitted to peripheral blood., (Copyright 2003 S. Karger AG, Basel)

The classical cytogenetic assay to estimate the dose to which an individual has been exposed relies on the measurement of chromosome aberrations in lymphocytes at the first post-irradiation mitosis 48 h after in vitro stimulation. However, evidence is accumulating that this protocol results in an underestimation of the cytogenetic effects of high LET radiation due to a selective delay of damaged cells. To address this issue, human lymphocytes were irradiated with C-ions (25-mm extended Bragg peak, LET: 60-85 keV/ micro m) and aberrations were measured in cells reaching the first mitosis after 48, 60, 72 and 84 h and in G2-phase cells collected after 48 h by calyculin A induced premature chromosome condensation (PCC). The results were compared with recently published data on the effects of X-rays and 200 MeV/u Fe-ions (LET: 440 keV/ micro m) on lymphocytes of the same donor (Ritter et al., 2002a). The experiments show clearly that the aberration yield rises in first-generation metaphase (M1) with culture time and that this effect increases with LET. Obviously, severely damaged cells suffer a prolonged arrest in G2. The mitotic delay has a profound effect on the RBE: RBE values estimated from the PCC data were about two times higher than those obtained by conventional metaphase analysis at 48 h. Altogether, these observations argue against the use of single sampling times to quantify high LET induced chromosomal damage in metaphase cells., (Copyright 2003 S. Karger AG, Basel)

We describe a peculiar and relatively rare type of chromosomal rearrangement induced in human peripheral lymphocytes that were ostensibly irradiated in G(0) phase of the cell cycle by accelerated heavy ions, and which, to the best of our knowledge, have not been previously described. The novel rearrangements which were detected using mFISH following exposure to 500 MeV/nucleon and 5 GeV/n 56Fe particles, but were not induced by either 137Cs gamma rays or 238Pu alpha particles, can alternatively be described as either complex chromatid-isochromatid or complex chromatid-chromosome exchanges. Different mechanisms potentially responsible for their formation are discussed., (Copyright 2003 S. Karger AG, Basel)

G(0) human blood lymphocytes were irradiated with 2.0 Gy gamma-rays and cultured to metaphase whilst held in a 50-Hz power frequency magnetic field of 0.23, 0.47 or 0.7 mT. No differences were found in the frequencies of gamma-induced chromosome aberrations observed in cells held in the EM fields compared with replicates held in a sham coil. Similar field conditions have been reported to increase the frequency of gamma-induced HPRT mutations, leading to a suggestion that the EM fields alter the fidelity of repair of genomic lesions. This was not confirmed by the chromosome aberration assay described here., (Copyright 2003 S. Karger AG, Basel)

Human peripheral lymphocytes were exposed to 137Cs gamma-rays (0-4.3 Gy) in order to check the impact of unstable cells on the dose-response curve for translocations. Chromosomes 2, 4 and 8 were FISH-painted. 17,720 first dividing cells were analysed. For the discrimination between stable and unstable cells the painted and the counter-stained chromosomes were analysed at doses of 1 Gy and higher. The cell distribution of translocations follows a Poisson distribution. The data were fitted to the linear-quadratic function, y = c + alphaD + betaD2. As expected, the alpha coefficients of the dose-response curves for translocations in stable cells or in total cells do not differ. However, at doses >1 Gy, the frequency of all translocations in stable cells seems to be lower than the frequency in total cells. For the establishment of calibration curves for past dose assessment purposes, only complete translocations should be scored, in order to estimate reliable doses., (Copyright 2003 S. Karger AG, Basel)

The incidence of spontaneous aneuploidy in human somatic and germ cells is known to be positively associated with aging. However, the influence of age on the individual susceptibility to chemically induced chromosome malsegregation has not been elucidated. In this study the spindle poison vinblastine (VBL) was used as a model compound to assess the influence of donor age on chemically induced chromosome malsegregation in cultured lymphocytes. Blood cultures from 20 female donors belonging to two different age groups (10 <30 years and 10 >50 years) were treated with VBL (7.5 ng/ml) from 43 h after mitogen stimulation until harvest at 60 h, i.e. during the time interval corresponding to G2/M. In order to block cytokinesis, cytochalasin B (6 microg/ml) was added to cultures at 44 h. For each donor the incidence of micronuclei, polyploidy and malsegregation (non-disjunction and loss) of chromosomes X and 8 was determined using fluorescence in situ hybridization with chromosome-specific centromeric probes. Both background incidence of micronuclei and spontaneous chromosome X non-disjunction were significantly elevated in older donors. Individual responses to VBL treatment showed wide interindividual variability, which was not significantly associated with the age of the donor. In both age classes chromosome X was more susceptible than chromosome 8 to both spontaneous and VBL-induced malsegregation. These results indicate that donor age has a limited influence on the aneugenic effects exerted by VBL in peripheral lymphocytes in vitro. Other factors have to be considered to account for the large interindividual variation in sensitivity to VBL challenge observed in this work.

Oral contraceptives are highly efficient and easily administered drugs; however, it must not be forgotten that they are composed of chemical substances which can be classified as potential carcinogens. Testing of a substance for genotoxicity represents a reliable approach both to evaluate the genetic hazard and to obtain information on its possible tumorigenic (cancerogenic) properties. The present study was undertaken to evaluate through carefully planned and controlled investigations the in vitro cytogenetic effects of oral contraceptives (ethynilestradiol and norgestrel mixed in the proportion 1:5) using three different concentrations, with two different durations of treatment (48 and 72 h), on two types of human cells (lymphocytes and fibroblasts) and a series of short-term test procedures: sister chromatid exchange (SCE), micronucleus test (MN), and chromosome aberrations (CA). In addition, the FISH procedure and in vitro anaphase and metaphase preparation analyses were performed. In contrast to CA and SCE frequencies, the frequency of MN in treated blood lymphocytes showed higher values by comparison with the controls, although the difference was statistically significant only for the lowest concentration (P = 0. 016). When using pancentromeric alphoid probes, the FISH procedure gave positive signals in more than 85% of micronuclei, clearly indicating that MN may contain whole chromosomes rather than acentric fragments. Unlike the lymphocytes, the fibroblasts showed dose-dependent effects, although those treated with the highest hormone concentrations showed an increased number of highly damaged cells (cytoplasmatic vacuolization, nuclear fragmentation, etc.), a decreased number of anaphase cells, a large number of which were abnormal, and a reduction of mitotic index. In conclusion, our data confirm that hormones do not induce structural chromosome aberrations in lymphocytes and indicate that ethynilestradiol and norgestrel have an aneugenic effect on fibroblast and lymphocyte cultures; FISH analysis on micronuclei from lymphocyte cultures and anaphase preparations from fibroblast cultures support this hypothesis. Teratogenesis Carcinog. Mutagen. 20:147-159, 2000., (Copyright 2000 Wiley-Liss, Inc.)

Effects of X-ray low doses (0.5-25 cGy) with following UV light (254 nm, 20 J/m2) on human peripheral blood lymphocytes were studied. Reparation response registered by unscheduled DNA synthesis (UDS) activity was demonstrated to be the most intensive after action of X-rays in dose ranges 2-3 cGy and 15-20 cGy, and least intensive after 10 cGy. In those cells where UV light was followed by X-rays, dose ranges 2-3 cGy and 15-20 cGy cause essential decrease of UDS as compared with UV-light action only. The most intensive UDS was in those UV-irradiated lymphocytes which where previously exposed to 10 cGy of X-rays. At the same time 10 cGy cause minimal reparation response without UV light. Possible mechanisms of discovered phenomena are discussed. In particular, reaction on 2-3 cGy might be a reorganization of genome for adaptive response or an evolution reserve cells response. From 10 cGy the reason of reaction might be a reparation, induced by radiation.

The paper presents results of investigation of constitutive heterochromatin of chromosomes 1, 9 and 16 in 23 patients with endometrial cancer and 5 women of the control group. The analysis was carried out on slides obtained by the routine method of the peripheral blood cell culture. The C-band segment variability was studied by the C-binding method. The investigation established an increase of polymorphism of constitutive heterochromatin in cancer patients, extreme variants and heteromorphism of homologues of chromosomes 1, 9 and 16. It is shown that polymorphism of the C-band on chromosomes 1, 9 and 16 in patients with cancer in their families increases as compared to the patients without cancer pathology in pedigrees.

The clastogenic effect of halothane anesthesia on peripheral blood lymphocytes from operated patients was studied. Chromosomal aberrations and sister chromatic exchanges in lymphocyte cultures of 7 patients were analyzed. Blood samples were obtained before the operative intervention, immediately after the effect of anesthesia was over and 24 hours later. To differentiate sister chromatids in the culture medium, on the 24th cultivation hour bromodeoxyuridine in final concentration 10 micrograms/ml was added. Increased incidence of structural chromosomal aberrations was recorded in lymphocyte cultures of patients after anesthesia--6.43 per cent cells with aberrations in the first post-halothane culture and 8.28 per cent in the second. Comparison of the cell affection index with the total number of affected cells revealed a continuous clastogenic effect pattern within the study period. Reliable evidence for induction of sister chromatid exchanges after the anesthesia was not found. The results were debated in the light of the possible mechanisms of clastogenicity of halothane anesthesia, the initial state of the patients and the consequences for them.

Blood samples were spiked with Na-24 to study the separate effect of this nuclide on the incidence of chromosomal aberrations in neutron irradiated blood samples. A delay of 96 h was allowed before cultivation, so the results of chromosomal aberration analysis could be compared with the results obtained by direct irradiation of blood samples with U-235 fission neutrons [7]. The absorbed dose was calculated using a simple conservative model. From the results obtained we can conclude that Na-24 alone was not the reason for the difference in the incidence of chromosomal aberrations between blood samples cultivated immediately after "in vitro" irradiation by U-235 fission neutrons and samples which were cultivated after 96 h storage.

Thermophilic clones of lymphoblastoid cell cultures Namalwa were generated and found to be capable of life and reproduction at a temperature of 60 degrees C. The reproductive dynamics, cytology, and ultrastructure of these clones were studied.

The results of cytogenetic examination of 51 patients with acute radiation disease caused by Chernobyl accident show interindividual variability in frequency and spectrum of chromosome aberrations in men with the same diagnosis, who were first examined 9-38 months after the accident. It is shown that possibility to reveal a cytogenetic marker of radiation exposure grows up in cases with severe disease.

Human peripheral lymphocytes (HPL) were used to study the synergistic effects of hyperthermia and X-irradiation. The effects on the chromosome aberration frequencies of different combinations of heat and radiation exposure at different cell cycle phases were analysed at the first mitosis after irradiation. When unstimulated HPL were heated after irradiation with 2 and 3 Gy, respectively, the chromosomal aberration frequencies were significantly higher than following radiation exposure alone. Heat treatment during different phases of the cell cycle and irradiation during the G2 phase led to an increase in the aberration frequencies when the cells were heat-treated not later than 16 h before radiation. Furthermore we show that the number of breaks increased linearly with the duration of hyperthermia when HPL were heated for periods of 1-15 min at 45 degrees C 32 h before irradiation at 48 h after stimulation. Heat treatment alone during any phase of the cell cycle did not induce chromosome aberrations. The increased aberration frequencies are probably the result of inhibited repair of radiation-induced DNA strand breaks.

Five continuous lymphoid suspension cell cultures were established by cultivation of lymphocytes from African green monkeys. The immunological typing revealed the mixed pattern of cultures consisting of B and T cell types. Two types of viruses: herpesvirus and retrovirus C-type were demonstrated by electron microscopy. Indirect immunofluorescence and molecular hybridization studies showed one of the viruses to be an EBV-related herpesvirus of green monkeys designated HVGM and the other to belong to H (S) TLV-1 group.

The method of the cytokinetic blocking was used to determine, in 18 healthy male donors aged 21-35 years, the frequency of binuclear cells with micronuclei that amounted to 1.1 +/- 0.2% (0.3-2.4%) on the average. The dependence of this parameter upon radiation dose (60Co-gamma radiation, 0.05 to 2 Gy) was a linear quadratic function and was described by the following regression equation: Yx = 0.76 + 5.76 D +/- 2.61 D2. With a dose of 0.05 Gy the frequency of binuclear cells with micronuclei exceeded significantly the control level.

A study was made of the dose dependence of the chromosome aberration frequency in human lymphocytes exposed to 60Co-gamma radiation and neutrons (mean energy of 0.85 MeV) at the G0 stage and in different periods of the G1 and G1/S stages of the cycle. With gamma irradiation the dose dependence for cells at the G1 and G1/S stages was at a higher level than that for cells at the G0 stage, whereas the opposite picture was observed for cells exposed to neutron radiation. The difference was also noted in the time-response curves where gamma radiation increased and neutrons, on the contrary, decreased the aberration yield in the cells that passed from G0 to G1 stage. The experimental data obtained are attributed to activation of repair system at the G1 stage which is mainly conditioned by chromatin decondensation; the activating, that is, the functional factor influences the aberration induction with gamma irradiation, while the decondensation, that is, the structural factor, with neutron irradiation.

The number of nucleoli was counted in the rabbit lymphocyte nuclei. This number was higher in meat rabbits than in skin-meat rabbits. We suppose that the polymorphism in the number of nucleoli reflecting the functional activity of ribosomal genes may be associated with biological-farming direction of productivity and energy of the animal growth.

The significant protective effect of recombinant interferon in the cultures of lymphocytes of healthy donors and patients with bronchial asthma has been revealed. The cytogenetic damage were stimulated by alkylating agents thioTEPA and photrin during their administration at the stages Gi-S of the cell cycle. No differences were revealed in the action of mutagens and protector in the patients and healthy persons.

The cytogenetic data obtained indicate a real possibility to intensify damage of the human cells as affected by rapid neurons. It can be achieved by combination of reparation inhibitors (caffeine) and rare-ionizing radiation (gamma rays) and is expressed in 5.5-fold increase of yield of chromosome damages in caffeine case and in increase of yield of exchange aberrations in the case of combination with gamma radiation.

Sister chromatid exchange (SCE) frequency at different times of fixation was studied in human lymphocyte cultures obtained from 6 donors. No differences were found in the SCE frequency between human lymphocyte cultures fixed at 72 and 96 hours of incubation (10.61 +/- 0.85 and 10.15 +/- 0.81 SCE per cell, respectively). However, a decreased SCE frequency (8.11 +/- 0.36 SCE per cell) was observed in cultures fixed at 120 hours of incubation. For a more detailed studies, one lymphocyte culture was fixed at different times of incubation (from 56 to 128 hours, at each a 8 hours). A slight increase in SCE frequencies was found at the interval between 56 and 88 hours of incubation, while starting from 104 hours of incubation a marked decrease in the SCE frequency was observed. Time-dependent changes in the SCE frequency may be described by the equation y = -1.8614 + 0.3922x - (2.5183 x 10(-3))x2, where y is the number of SCEs per cell, and x--the duration of culture incubation in hours. The observed phenomenon may be associated with changes in proportion of T and B lymphocytes, or with heterochromatization of chromosomes during a prolonged cultivation, or with an early in vitro stimulation of the in vivo long-lived lymphocytes that may be more damaged than the in vivo short-lived and the in vitro late-stimulating ones.

The observation of heritable fragile sites on human chromosomes prepared for lymphocyte cultures has been shown to depend on the type of tissue culture medium in which the lymphocytes are grown. The sites are observed at a much greater frequency when medium 199 is used than when RPMI 1640, Ham's F10, Eagle's (basal), and CMRL 1969 are used. One site on the X chromosome is of clinical significance in that it is a marker for X-linked mental retardation.

Human lymphocytes and Chinese hamster ovary (CHO) cells in culture were exposed for 12 1/2 hours to a magnetic resonance imaging apparatus with a 2.35-Tesla magnet and 100-MHz radio frequency emission. The cells were examined for cytogenetic damage manifested either as chromosome aberrations or sister chromatid exchanges (SCEs), which constitute very sensitive measures of genetic and cellular damage. In either unstimulated or stimulated human lymphocytes, as well as in exponentially growing CHO cells, no increase in either chromosome aberrations or SCEs was found as a result of exposure to these MR conditions. The data indicate that long-term exposure to MR imaging conditions far exceeding those to be found in the clinical situation does not cause cytogenetic damage.