[Cytological and cytogenetic damages of spermatogonial cells in mice in the acute and late periods after irradiation by protons, helium ions, and gamma-rays].

Experiments with small test animals have been carried out to study the survival rate of spermatogenic cells in the acute period after exposure to protons with the energy 9 GeV, helium ions with the energy 4 GeV/nucleon, and 60Co gamma radiation in doses of 0.5-7.5 Gy and the reciprocal translocation frequency in spermatocytes under meiosis at the diakinesis-metaphase-1 stage six months after the exposure of the animals to protons with the energy 50 MeV and 9 GeV, helium ions with the energy 4 GeV/nucleon, and 60Co gamma-radiation in doses of 0.5-4.0 Gy. It is shown that the dependence of the effect on the dose is linear or near to linear for all kinds of radiation used. Relative biological effectiveness coefficients of the accelerated nuclei obtained by correlating equally effective doses of the standard and investigated radiations turned out to be higher in survival rate of type B spermatogonium and amounted to 2.0 and 1.3 for 9 GeV protons and helium ions respectively. At the same time, the use of the nonparametric method to determine the RBE coefficients in the course of finding out the reciprocal translocation frequency in spermatocytes points to an increase in the RBE coefficients of charged particles from 1.0 to 2.0.

Genetic control of susceptibility to pulmonary infection with Chlamydia pneumoniae in the mouse.

A mouse model was used to study the genetic control of differential host response to pulmonary infection with Chlamydia pneumoniae. The A/J and C57BL/6 strains show differential response to intranasal infection with respect to their ability to clear pulmonary bacterial load and the extent of lung pathology developed by 2 weeks post infection. The genetic basis of this interstrain difference was studied by whole-genome scan in an informative [A/J x C57BL/6J] F2 cross using the pulmonary microbial load as a phenotypic readout of host response. We detected a highly significant linkage (LOD score=11.5) on chromosome 17 that overlaps with the major histocompatibility (MHC) locus. This quantitative trait locus (QTL) accounts for approximately 30% of the phenotypic variance with B6 alleles conferring susceptibility and inherited in a recessive fashion. Significant linkage was also detected to chromosome 5 in female mice, while chromosome 6 showed suggestive linkage in male mice, pointing to additional complexity in the genetic control of the difference in susceptibility observed in A/J and C57BL/6J.

Bovine Chlamydophila spp. infection: do we underestimate the impact on fertility?

Classical methods for detection of Chlamydophila species, and of antibodies against these agents, have indicated that these bacteria are highly prevalent in cattle and associated with numerous disease conditions. These methods demonstrated acute Chlamydophila-induced diseases such as epizootic bovine abortion, as well as worldwide variable, but generally high, Chlamydophila seroprevalence. However, it was impossible to consistently detect the low levels of these organisms which were suspected to be present in endemic infections. Application of highly sensitive real-time PCR and ELISA methods for detection of Chlamydophila spp. DNA and of antibodies against Chlamydophila spp., respectively, in a series of prospective cohort studies revealed a high prevalence of Chlamydophila spp. genital infections in female calves (61%) and adult heifers (53%). These infections were acquired by extragenital transmission in the first weeks of life, and infection frequency was increased by crowding of the animals. A challenge study demonstrated that infection with C. abortus resulted in decreased fertility of heifers. The experimental use of a C. abortus vaccine provided evidence for immunoprotection against C. abortus-induced suppression of bovine fertility. The results of these investigations suggest that bovine Chlamydophila infection should be viewed more as pervasive, low-level infection of cattle than as rare, severe disease. Such infections proceed without apparent disease or with only subtle expressions of disease, but potentially have a large impact on bovine herd health and fertility.

Occupational exposure to lead and induction of genetic damage.

To investigate whether occupational exposure to lead is genotoxic, we evaluated data from 103 lead-exposed workers and 78 matched controls. These data correspond to three different sampling periods, and we measured genetic damage as increases in the frequency of binucleated cells with micronuclei (BNMN) in peripheral blood lymphocytes. The levels of exposure were determined according to the lead levels in blood. Clearly significant increases in BNMN were observed in the exposed groups when compared to the control group. In addition, for the overall population (n = 181), we observed a clear relationship between lead levels in blood and BNMN (r = 0.497; p < 0.001). When we examined four exposure levels--very low exposure (< 1.20 microM/L), low exposure (1.20-1.91 microM/L), high exposure (1.92-2.88 microM/L), and very high exposure (> 2.88 microM/L)--we found significant differences in the genetic damage induction. We conclude that exposure to levels of lead higher than 1.20 microM/L may pose an increase in genetic risk. In addition, our data show that blood lead level is a good indicator of genetic damage induction.

Genotoxicity and radioresistance in electroplating workers exposed to chromium.

A biomonitoring study was carried out to investigate the genetic risk associated to occupational exposure to chromium. The induction of genetic damage was measured by analysing the frequency of micronuclei (MN) in peripheral blood lymphocytes. In addition to the 40 electroplater exposed workers who participated in the study, a group constituted by 18 volunteer donors, without exposure to chromium, was analysed as a control group. Measures of chromium levels at working place and in erythrocytes and urine were obtained, as indicators of exposure. The results from this study indicate that the blood from exposed workers contained higher levels of chromium, when compared with those obtained in the control group, and that a significant increase in the frequency of both the total number of MN and the number of binucleated cells carrying MN (BNMN) was detected. Furthermore, a good direct relationship was obtained between the amount of chromium present in air, erythrocytes or urine and the frequency of MN. To determine the existence of radioresistance as consequence of chromium exposure, the response of lymphocytes to the in vitro gamma-radiation was studied. The results of this experiment show a lower induction in the increase of the frequency of MN after challenge irradiation in the lymphocytes of chromium exposed workers, which should be indicative of an adaptive response.

Evaluation of DNA damage by the Comet assay in shoe workers exposed to toluene and other organic solvents.

The alkaline single-cell gel electrophoresis (or Comet) assay was applied to evaluate DNA damage in cryopreserved peripheral blood mononuclear leukocytes from 34 female shoe workers exposed to organic solvents and a group of 19 non-exposed women. We also investigated whether the polymorphisms of glutathione S-transferase M1 (GSTM1) and T1 (GSTT1) genes affect individual level of DNA damage possibly induced by the solvent exposure. Chemical measurements of workplace air in the two factories studied showed that the workers were exposed to acetone, gasoline, and toluene in both factories and to ethylacetate and diisocyanate in one factory. In the exposed workers, the average level of blood hemoglobin was lower and that of urinary hippuric acid higher than in the non-exposed individuals. However, the occupational exposure to organic solvents did not affect the Comet values. Neither did age, smoking, or the GSTM1 genotype have any effect on the outcome of this assay. The low prevalence of the GSTT1-null genotype precluded conclusions on the influence of GSTT1 polymorphism.

Biomonitoring of workers exposed to lead. Genotoxic effects, its modulation by polyvitamin treatment and evaluation of the induced radioresistance.

A population monitoring study was performed, by using the micronucleus (MN) assay in human peripheral lymphocytes, to investigate whether occupational exposure to lead is genotoxic to workers. In addition to the exposed workers group, two more groups were studied, an external group from a factory without exposure to lead and an internal control group, from the same factory as the exposed workers, but that were not directly exposed to lead. Measures of lead levels at working place and in blood were calculated, and blood samples were collected to carry out a MN study. The results from these studies indicate that the blood from workers directly exposed contained high levels of lead, compared with the other groups, and a significant increase in the frequency of both the total number of MN and the number of binucleated cells carrying MN appeared. In addition, a study on the antimutagenic effects of a polyvitamin rich diet was conducted by measuring the frequency of MN after the workers had a four month daily intake of a polyvitamin-polymineral complex. These results clearly show a significant reduction of the MN frequency evaluated after this treatment, obtaining values that were even lower than those obtained in the internal control group. Finally, a challenge assay was carried out to determine response to gamma-radiation as indication of any kind of radiosensitivity or radioresistance. The results of this experiment did not show any significant variation in the increase of the frequency of MN after challenge irradiation in the lead exposed workers; nevertheless this increase was significantly reduced in the sample obtained after the polyvitamin treatment indicating a radioresistance response.