Cytogenetic Analysis Of Peripheral Lymphocytes Research Articles

Chromosome analysis of nuclear power plant workers using fluorescence in situ hybridization and Giemsa assay

The aim of this study was to evaluate the genotoxic effects of ionizing radiation in vivo in exposed Bulgarian nuclear power plant workers by using classical cytogenetic and molecular cytogenetic analyses of peripheral lymphocytes. Chromosome analysis using fluorescence in situ hybrydization (FISH) and Giemsa techniques was undertaken on 63 workers and 45 administrative staff controls from the Bulgarian Nuclear Power Plant. Using the Giemsa method, the frequencies of cells studied with chromosome aberrations, dicentrics plus rings and chromosome fragments in the radiation workers were significantly higher compared with the control group (P = 0.044, P = 0.014, and P = 0.033, respectively). A significant association between frequencies of dicentrics plus rings and accumulated doses was registered (P < 0.01). In the present study, a FISH cocktail of whole chromosome paints for chromosomes 1, 4 and 11 was used. A significant association between frequency of translocations and accumulated doses was also observed (P < 0.001). Within the control group, a correlation was found between age and the spontaneous frequency of translocations. No correlation was found between smoking status and frequency of translocations. When compared with the control group, workers with accumulated doses up to 100 mSv showed no increase in genome translocation frequency, whereas workers with accumulated doses from 101 to 200 mSv showed a statistically significant doubling of genome translocation frequency (P = 0.009). Thus, in cases of chronic exposure and for purposes of retrospective dosimetry, the genome frequency of translocations is a more useful marker for evaluation of genotoxic effects than dicentric frequency.

Nonmosaic 47,XYY syndrome presenting with male infertility: case series

In this study, we describe nine patients with 47,XYY presenting with male infertility. All patients were subjected to history taking, clinical examination, duplex ultrasonographic examination of the scrotum, endocrinological investigations and cytogenetic analysis of peripheral lymphocytes. Two patients tried intracytoplasmic sperm injection (ICSI). Our results showed that seven patients were oligospermic and two patients were azoospermic. Bilateral varicocele was detected in seven patients. The hormonal levels in the majority of the patients were within normal range. Two patients showed improvement after varicocelectomy. The wife of one of the oligospermic patients became successfully pregnant after the first trial of ICSI. In conclusion, this report suggests that patients with XYY may present with primary infertility and may show oligospermia and nonobstructive azoospermia. Careful clinical, ultrasonographic, endocrinological and cytogenetic examinations should be a part of their diagnostic work-up for the proper management of these patients. In addition, ICSI may be a hope for some of these patients.

Human biomonitoring system in the Czech Republic

The human biomonitoring (HBM) is an integral part of Environmental Health Monitoring System in the Czech Republic since 1994. Selected biomarkers of the internal dose (heavy metals, PCBs) and cytogenetic analysis of peripheral lymphocytes as a biomarker of the exposure/effect to/of environmental genotoxic factors are systematically followed up in the blood and urine of adults (blood donors), in children aged 8 to 10 years, and in the breast milk of nursing mothers. Selected outputs documented the declining trend of blood lead levels, with the recent reference value of 80 mg/l for men, and the rising trend of blood selenium levels in adults, but not in children. PCBs and chlorinated pesticides in human milk show a long-term downward trend, but still higher than in neighbouring countries. The frequency of aberrant cells revealed a downward trend, but the increase obtained in the last monitored period needs to be explained. Further HBM activities are required to demonstrate the corresponding trends and to reduce human exposure and health risks.

Intrachromosomal triplication for the distal part of chromosome 15q

We report the case of a boy whose karyotype at birth showed additional material on one chromosome 15. He underwent treatment for unilateral nephroblastoma at 6 years old. At 23 years old, he presented with body asymmetry, facial dysmorphism, arachnodactyly, severe scoliosis, and mental retardation. Molecular cytogenetic analyses of peripheral lymphocytes demonstrated a complex mosaic with three clones. A major cell lineage (68%) showed a chromosome 15 with additional material fused to its telomere long arm that was constituted by an inverted duplicated 15q24.3-qter segment. Therefore, the resulting add(15)(q) harbored an intrachromosomal triplication with the middle segment being inverted in orientation. A minor cell lineage (7%) showed an abnormal chromosome 3 resulting from a telomeric fusion between its short arm and an inverted duplicated 15q24.3-qter segment. The third cell lineage (25%) showed a normal 46,XY constitution. Finally, this resulted in tetrasomy for the distal 15q24.3-qter region in 75% of the patient's lymphocytes. To our knowledge, distal 15q tetrasomy is rare and only eight cases have been reported in the literature, all due to a supernumerary analphoid marker consisting of an inverted duplication. We report here the first observation of distal 15q tetrasomy associated with a 46 chromosomes constitution. We compare the phenotype of our patient to previous cases of distal tetrasomy 15q and discuss the mechanisms underlying this chromosomal rearrangement.

A novel case of infantile sacral teratoma and a constitutional t(12;15)(q13;q25) pat

Cytogenetic analysis of peripheral lymphocytes of an infantile patient with a sacral teratoma revealed a constitutional translocation (12;15)(q13;q25) pat. The same translocation was found in four additional relatives. Loss of heterozygosity analysis of the patient's tumor material showed retention of both translocation-derived chromosomes. Since allelic loss in the 12q13 region has been observed in germ cell tumors, we hypothesize that disregulation of genes located at or near the 12q13 breakpoint may be related to the development of this sacral teratoma. As a first step towards the identification of these genes, a 12q13 genomic contig that spans the breakpoint has been constructed.

Cytogenetic effects of hexavalent chromium in Bulgarian chromium platers

The aim of the present study was to evaluate the genotoxic effects of hexavalent chromium (Cr(VI)) in vivo in exposed Bulgarian chromium platers by using classical cytogenetic and molecular cytogenetic analyses of peripheral lymphocytes and exfoliated buccal cells. No significant difference was observed between the exposed workers and the controls with regard to the frequency of cells with chromosome aberrations (CAs) using conventional Giemsa staining and in the frequency of sister chromatid exchanges (SCEs). However, there was a significant increase in the number of cells with micronuclei (MN) in peripheral lymphocytes from chromium exposed workers as compared to the controls. In the buccal cells from these workers, this increase was even more pronounced. Cytosine arabinoside (AraC), an inhibitor of DNA synthesis and repair, was found to significantly increase the levels of MN in vitro in the lymphocytes of both groups. The increase was more expressed in the lymphocytes of chromium exposed workers. Both centromere positive (C +) as well as centromere negative (C −) MN were observed by the fluorescence in situ hybridization (FISH) technique in both of the cell types studied. No difference between C + and C − MN frequencies was found in the lymphocytes as well as in the buccal cells. Thus, Cr(VI) appears to have both clastogenic as well as aneugenic effects in humans.

Frequency of HPRT mutants in humans exposed to vinyl chloride via an environmental accident

The mutant frequency (MF) in the hypoxanthine-guanine-phosphoribosyl-transferase (HPRT) locus of peripheral blood T-lymphocytes was measured in a population environmentally exposed to vinyl chloride — a toxic and carcinogenic substance through an accidental release into the atmosphere. It was compared to MF in a control group of unexposed individuals. Both groups were re-investigated in a follow-up study, 2 years later. No significant difference could be observed in MF between exposed and controls either at the accident nor in the follow-up study. Approximately the same mean HPRT mutant frequencies were observed for both groups in T-lymphocytes from blood samples obtained shortly after the accident and from the follow-up blood samples. Both groups showed a higher mean MF in the re-investigation samples which is most probably due to the significantly lower average cloning efficiency (CE) under non-selective conditions and because of the inverse relationship between CE and MF. The exposed population showed a higher mean T-cell CE at the initial blood sampling as compared to the control group. The concurrent cytogenetic analyses of peripheral lymphocytes showed a significant increase in cells with aberrations in the exposed population. Clastogenic but not mutagenic activity of vinyl chloride was observed in our study.

Variable levels of mosaicism for trisomy 21 in a non-immune hydropic infant with chylothorax

We report the first case of mosaic trisomy 21 with non-immune hydrops fetalis and bilateral chylothoraces. Prenatal fetal blood karyotype analysis of 15 fetal cells revealed a 46,XX karyotype. Aggressive prenatal management, including fetal thoracocentesis and pleuro-amniotic shunt, was performed. A clinical phenotype of Down syndrome was apparent after the gross oedema had subsided. Subsequent chromosome study of neonatal blood lymphocytes showed mosaic trisomy 21 with 23 per cent trisomic cells. Review of the initial fetal blood sample identified trisomy in 5 per cent of 134 cells. Follow-up study at five months showed no trisomy 21 in 100 cells. This case illustrates the variable levels of mosaicism manifest in the peripheral blood of an infant with obvious Down syndrome phenotype, and the limitation of cytogenetic analysis of peripheral lymphocytes alone in prenatal and postnatal detection of low levels of mosaicism.

Analysis of the sex chromosome constitution of sperm in men with a 47,XYY mosaic karyotype by fluorescence in situ hybridization

Objective: To determine the incidence of sex chromosome aneuploidy in the sperm of two men with a 47,XYY/46,XY karyotype. Design: Case report. Setting: Infertility clinic in a teaching hospital. Patient(s): One patient with near normal semen parameters whose wife had a history of miscarriages and one patient with primary infertility and severe oligoasthenozoospermia. Intervention(s): Cytogenetic analysis of peripheral lymphocytes and three-color X/Y/18 fluorescence in situ hybridization analysis of sperm. Main Outcome Measure(s): Analysis of sex chromosome disomy and diploidy rates in sperm. Result(s): Both patients had a 47,XYY/46,XY karyotype. The hyperdiploidy rate of patient 1 was 19% and that of patient 2 was 90%. The incidence of disomy XY was significantly elevated in both patients compared with the controls (0.23% and 1.02%, respectively, versus 0.10%). The incidence of disomy YY (0.44% versus 0.10%) was increased only in patient 2, as was the incidence of disomy 18 (0.49% versus 0.09%) and the rate of diploidy (0.83% versus 0.13%). The rate of 24,XX sperm in both patients was not different from that in the controls. Conclusion(s): Patients with a 47,XYY mosaic karyotype may be at risk of producing offspring with a hyperdiploid sex constitution. These patients should have their sperm investigated by fluorescence in situ hybridization to determine their particular risks before they undergo intracytoplasmic sperm injection.

Cytogenetic analysis of peripheral lymphocytes in a population exposed to vinyl chloride through an accidental release into the environment

On the first of June, 1996 an environmental accident occurred in Schönebeck, Germany in which free vinyl chloride was evaporated into the atmosphere. Thereby, the human population living in this area was exposed to vinyl chloride and its byproducts. Chromosomal aberrations were measured in peripheral blood lymphocytes from 29 potentially exposed and 29 non-exposed (control) individuals. Both groups were matched regarding age, gender and smoking habits. Two hundred metaphases were analysed for chromosomal aberrations per each individual. The exposed group showed a statistically significant increase in the mean frequency of aberrant cells (1.47% versus 1.07% in the control group). Chromosomal aberrations in peripheral lymphocytes have been shown to be a very sensitive biomarker of genotoxic effects not only for occupational exposure to vinyl chloride as reported several times during the last 20 years, but also for an accidental environmental exposure. A follow-up cytogenetic study is recommendable.

Cytogenetic analysis of peripheral lymphocytes in medical personnel by means of FISH

Genotoxic effects of occupational exposure to cytostatics were investigated in 20 nurses and physicians working in various departments of one hospital. The group was divided into two equal subgroups one of which was involved in the administration of cytostatics (exposed subgroup) and the other was not (unexposed subgroup). The whole group and the two subgroups were compared with a control group of 11 healthy blood donors. Two differently labeled whole chromosome painting (WCP) probes specific for the chromosomes 1 and 4 were used simultaneously. Chromosome aberrations were classified in terms of the Protocol for Aberration Identification and Nomenclature (PAINT) nomenclature. The results obtained by the painting method were compared with findings of conventional unbanded chromosome analysis. Significant differences in the numbers of translocations ( F G/100=2.25±1.50 vs. 0.66±0.21, p<0.01) and unstable chromosome aberrations determined by the conventional method (AB.C/100=2.70±2.31 vs. 1.63±1.59, p<0.05) were found between the exposed subgroup and controls. The unexposed subgroup differed from the controls only in the number of translocations ( F G/100=2.93±2.79 vs. 0.66±0.51. p<0.05). No significant differences in the number of stable and unstable aberrations were found between the exposed and the unexposed subgroups. On the other hand, highly significant differences ( p<0.01) were demonstrated by the two methods between the whole group (all medical personnel) and the controls. All differences which were found to be significant when translocations were compared were also found to be significant when total stable chromosome exchanges, i.e., the sum of translocations and insertions, were considered. Multicolour chromosome painting is apparently a more sensitive method than the conventional metaphase-based analysis.

Population-based biomonitoring in the Czech Republic — the system and selected results

In the framework of the system of monitoring the environmental impact on population health, the concentration of lead, cadmium and selenium in blood and cadmium in urine was measured in adults ( n = 670), children ( n = 599) and umbilical blood ( n = 549) using atomic absorption spectrophotometry. Furthermore, cytogenetic analysis of peripheral lymphocytes in all population groups under study was investigated. The median blood Pb level for the overall group of adults (47.8 μg/l, i.e. 0.23 μmol/l) was significantly higher in men (51.5 μg/l, i.e. 0.25 μmol/l). Smoking significantly influenced the blood Pb level in women. The 90th percentile in no group exceeded the value of 100 μg/l (0.48 μmol/l). The median blood Cd level in adults (0.9 μg/l, i.e. 0.008 μmol/l) depends on smoking habit (1.25 μg/l, i.e. 0.01 μmol/l). The median urine Cd level was 0.585 μg/g creatinine (0.59 μmol/mole creatinine) in adults and 0.37 μg/g creatinine (0.37 μmol/mole creatinine) in children. The median blood Se level (53.5 μ/l, i.e. 0.68 μmol/l) was found to be higher in the group of non-smokers (57.5 μg/l, i.e. 0.73 μmol/l). Lead and selenium level were significantly lower in the umbilical blood. Cytogenetic analysis results showed age-dependent average percentages of aberrant cells: 1.1% in umbilical blood, 1.27% in children and 1.71 in adults in line with the reference values for the Czech population.

P XVI B.2 - P XVI B.2 Cytogenetic analysis of peripheral lymphocytes in health personnel by means of fish

P XVI B.2 - P XVI B.2 Cytogenetic analysis of peripheral lymphocytes in health personnel by means of fish

Chromosome investigations in weightlifters and powerlifters after intake of anabolic steroids

A total of 231 weightlifters and powerlifters were asked in an anonymous poll whether they had taken anabolic steroids (AS) for longer than 3 months. A sample of blood was taken from the 17 athletes (7.4%) who answered positively and agreed to cytogenetic analysis of peripheral lymphocytes in order to identify the possible genotoxic risk, and the results were compared with a reference group of 45 blood donors. On the basis of cytogenetic analysis, the increased quantity of chromosome aberrations in peripheral lymphocytes was found to be 2.6% in comparison with the level of 1.3% in a reference group. These results were also verified by the distribution of aberrant cells in cultures of lymphocytes that were analyzed. These results indicated an increased risk of genotoxic exposure of these athletes, which might cause an increase in malignant diseases.

Chromosome and sister-chromatid exchange analysis in peripheral lymphocytes, and mutagenicity of urine in anesthesiology personnel.

Genotoxic effects of occupational exposure to anesthetic gases (halothane and nitrous oxide) were studied in 24 persons working at departments of anesthesiology and resuscitation (14 anesthesiologists and 10 anesthesiologic nurses). Frequencies of chromosomal aberrations were determined in 48-h cultures, and sister-chromatid exchanges (SCEs) in 72-h cultures of peripheral lymphocytes. In parallel, the mutagenetic potential of urine was tested using Ames tester Salmonella typhimurium strains TA98 and TA100 in both the presence and the absence of a metabolic activation system. The control group consisted of 30 healthy blood donors from the Clinic of Hematology and Blood Transfusion. Halothane concentrations in the working environment of operating rooms were determined using gas-liquid chromatography. Halothane exposure levels were far above the maximum allowable concentration in most countries, ranging between 9 and 490 mg.m-3. Cytogenetic analysis of peripheral lymphocytes revealed significantly increased frequencies of aberrant cells (ABCs) and SCEs in the exposed group as compared with the controls (3.84% vs 1.87% ABCs, 8.69 +/- 1.30 vs 7.86 +/- 1.07 SCEs per cell). Results of the urine mutagenicity test were negative.

Mutagenicity studies on paracetamol in human volunteers. I. Cytogenetic analysis of peripheral lymphocytes and lipid peroxidation in plasma

The clastogenic activity of paracetamol (PC) was assayed on a group of 11 healthy volunteers. PC was administered in the form of tablets 3 × 1000 mg in the course of 8 h. Blood samples were taken 0, 24, 72 and 168 h after the first application of the drug. Each blood sample was used for the cytogenetic analysis of peripheral lymphocytes, for the measuring of the level of lipid peroxidation (LPO) and ascorbemia in plasma. After PC administration the frequency of aberrant cells (AB.C.) was increased to 2.77% AB.C. after 24 h vs. 1.68% at 0 h, and breaks per cell (B/C) to 0.0295 vs. 0.0182, respectively. If PC was applied simultaneously with ascorbic acid (AA), also in a dose of 3 × 1000 mg, an increased frequency of AB.C. was observed only after 72 h, of B/C after both 24 h and 72 h. No increase in LPO as determined by the thiobarbituric acid assay was seen after PC administration (1.02−1.10 nmole malondialdehyde (MDA)/ml plasma). The LPO level was increased 72 h after the simultaneous application of PC and AA (1.26 nmole MDA/ml). No effect of AA in terms of a decreased PC clastogenicity was observed.

Cytogenetic analysis of peripheral lymphocytes in workers occupationally exposed to soft-coal pressure gasification

Cytogenetic analysis of peripheral lymphocytes in workers occupationally exposed to soft-coal pressure gasification

Cytogenetic analysis of peripheral lymphocytes of workers occupationally exposed to styrene

Cytogenetic analysis of peripheral lymphocytes of workers occupationally exposed to styrene

The use of the cytogenetic analysis of peripheral lymphocytes as a method for checking the level of MAC in Czechoslovakia

The use of the cytogenetic analysis of peripheral lymphocytes as a method for checking the level of MAC in Czechoslovakia

Cytogenetic analysis of peripheral lymphocytes in workers occupationally exposed to epichlorohydrin.

Peripheral lymphocytes of 3 groups of adult subject (28 workers occupationally exposed for 4 years to epichlorohydrin [ECHH], 34 matching controls and 21 subjects of the general population (control group) were cytogenetically analysed. In total, 11 806 metaphases were scored. The following frequencies of aberrant cells (bearing mostly chromatid and chromosome breaks) were detected in individual groups: ECHH-exposed group 3.12%; matching controls 2.06%; general population control group 1.33%. The results seem to indicate that even the concentration of 1 mg ECHH/m3 was capable of increasing the frequency of aberrant cells in the occupationally exposed group of workers.