Oto Osina

Chromosomal damage among medical staff occupationally exposed to volatile anesthetics, antineoplastic drugs, and formaldehyde.

OBJECTIVES Structural chromosomal aberrations in blood lymphocytes represent a biomarker for cellular damage caused by genotoxic carcinogens and are an indicator of increased cancer risk. We evaluated the association between frequencies of total chromosomal aberrations, chromatid- and chromosome-type aberrations, and occupational exposures to volatile anesthetics, antineoplastic agents, and formaldehyde among 601 medical professionals. METHODS Chromosomal damage among exposed individuals and unexposed controls was determined by conventional cytogenetic analysis. We used binary logistic regression to evaluate the effects of workplace exposures and major confounders on chromosomal damage. RESULTS Significantly higher frequencies of total chromosomal, chromatid-type and chromosome-type aberrations were observed among subjects occupationally exposed to volatile anesthetics, antineoplastic agents, and formaldehyde compared to age- and sex-matched controls (P<0.0001). The risk of an increased frequency of chromosomal aberrations was associated with exposure to anesthetics [odds ratio (OR) 3.9, 95% confidence interval (95% CI) 2.7-5.8], cytostatics (OR 2.7, 95% CI 1.9-3.9), and formaldehyde (OR 1.7, 95% CI 1.1-2.7). No other covariate contributed significantly to the model. Chromatid- and chromosome-type aberrations were associated with exposure to anesthetics and cytostatics without any contribution of other variables. Stratified data analysis showed the risk of increased chromosomal aberrations among non-smoking female nurses and physicians exposed to anesthetics, cytostatics and, partially, formaldehyde. Chromatid and chromosome exchanges were significantly higher in the exposed groups than among controls. CONCLUSION Our findings indicate that the presence of genotoxic compounds in operating rooms, oncological units, and pathological departments results in a significant increase of chromosomal damage (impair of chromosomal integrity) among medical workers employed in these facilities.

Chromosomal aberrations in tire plant workers and interaction with polymorphisms of biotransformation and DNA repair genes

We evaluated chromosomal aberrations in lymphocytes of 177 workers exposed to xenobiotics in a tire plant and in 172 controls, in relation to their genetic background. Nine polymorphisms in genes encoding biotransformation enzymes and nine polymorphisms in genes involved in main DNA repair pathways were investigated for possible modulation of chromosomal damage. Chromosomal aberration frequencies were the highest among exposed smokers and the lowest in non-smoking unexposed individuals (2.5+/-1.8% vs. 1.7+/-1.2%, respectively). The differences between groups (ANOVA) were borderline significant (F=2.6, P=0.055). Chromosomal aberrations were higher in subjects with GSTT1-null (2.4+/-1.7%) than in those with GSTT1-plus genotype (1.8+/-1.4%; F=7.2, P=0.008). Considering individual groups, this association was significant in smoking exposed workers (F=4.4, P=0.040). Individuals with low activity EPHX1 genotype exhibited significantly higher chromosomal aberrations (2.3+/-1.6%) in comparison with those bearing medium (1.7+/-1.2%) and high activity genotype (1.5+/-1.2%; F=4.7, P=0.010). Both chromatid- and chromosome-type aberration frequencies were mainly affected by exposure and smoking status. Binary logistic regression analysis revealed that frequencies of chromatid-type aberrations were modulated by NBS1 Glu185Gln (OR 4.26, 95%CI 1.38-13.14, P=0.012), and to a moderate extent, by XPD Lys751Gln (OR 0.16, 95%CI 0.02-1.25, P=0.081) polymorphisms. Chromosome-type aberrations were lowest in individuals bearing the EPHX1 genotype conferring the high activity (OR 0.38, 95%CI 0.15-0.98, P=0.045). Present results show that exposed individuals in the tire production, who smoke, exhibit higher chromosomal aberrations frequencies, and the extent of chromosomal damage may additionally be modified by relevant polymorphisms.

Interactions of DNA repair gene variants modulate chromosomal aberrations in healthy subjects

Human cancers are often associated with numerical and structural chromosomal instability. Structural chromosomal aberrations (CAs) in peripheral blood lymphocytes (PBL) arise as consequences of direct DNA damage or due to replication on a damaged DNA template. In both cases, DNA repair is critical and inter-individual differences in its capacity are probably due to corresponding genetic variations. We investigated functional variants in DNA repair genes (base and nucleotide excision repair, double-strand break repair) in relation to CAs, chromatid-type aberrations (CTAs) and chromosome-type aberrations (CSAs) in healthy individuals. Chromosomal damage was determined by conventional cytogenetic analysis. The genotyping was performed by both restriction fragment length polymorphism and TaqMan allelic discrimination assays. Multivariate logistic regression was applied for testing individual factors on CAs, CTAs and CSAs. Pair-wise genotype interactions of 11 genes were constructed for all possible pairs of single-nucleotide polymorphisms. Analysed individually, we observed significantly lower CTA frequencies in association with XPD Lys751Gln homozygous variant genotype [odds ratio (OR) 0.64, 95% confidence interval (CI) 0.48-0.85, P = 0.004; n = 1777]. A significant association of heterozygous variant genotype in RAD54L with increased CSA frequency (OR 1.96, 95% CI 1.01-4.02, P = 0.03) was determined in 282 subjects with available genotype. By addressing gene-gene interactions, we discovered 14 interactions significantly modulating CAs, 9 CTAs and 12 CSAs frequencies. Highly significant interactions included always pairs from two different pathways. Although individual variants in genes encoding DNA repair proteins modulate CAs only modestly, several gene-gene interactions in DNA repair genes evinced either enhanced or decreased CA frequencies suggesting that CAs accumulation requires complex interplay between different DNA repair pathways.

Cyclin D1 splice site variant triggers chromosomal aberrations in healthy humans.

Chromosomal aberrations (CAs) were suggested to be the cause of cancer by Boveri already a 100 years ago, but the detection of the Philadelphia chromosome about 50 years later provided the first direct evidence on the relationship (reviewed by Mitelman). CAs include recurrent aberrations, including specific translocations, deletions and amplifications of chromosomal regions or whole chromosomes. The detection of recurrent CAs was greatly facilitated when fluorescence in situ hybridization techniques were developed for interphase chromosomes. Non-specific CAs include missing, fragmented or fused chromosomal segments, divided into chromosome-type aberrations (CSAs) and chromatidtype aberrations (CTAs). These have been conventionally analyzed by microscopic scoring of metaphase nuclei from cultured lymphocytes. Modern sequencing techniques have revealed numerous gene fusions and rearrangements, in the extreme, chromothripsis, a catastrophic parsing of illegitimate chromosomal segments together. Non-specific CAs may arise as a result of direct DNA damage by, for example, ionizing radiation or replication on a damaged DNA template; the former lesions would be detected as CSAs, whereas the latter may be CSAs or CTAs. Non-specific CAs remain in lymphocytes for their lifetime. They have been used in monitoring of radiation exposure and exposure to genotoxic compounds and, together with sister chromatid exchanges and micronuclei, CAs have offered the only available method for human biomonitoring for genotoxic exposures. There are convincing data on the association of CA frequency with subsequent risk of many cancers. Furthermore, patients with many types of cancer show increased frequencies of CAs at diagnosis. CAs were assumed to be somatic events but our recent study on multiple myeloma showed that the recurrent t(11;14) (q13;q32) translocation was strongly associated with the cyclin D1 (CCND1) G870A genotype at a splice site. However, no effect was found for any other translocations or other CAs in myeloma. Here, we wanted to test the possible dependence of non-specific CAs on the CCND1 genotype by assaying for the G870A genotype in a large cohort of persons for whom CAs were measured. This polymorphism has been implicated as a risk factor for a number of cancers. The subjects (N1⁄4 731) were identified from occupational settings, including persons with defined exposures (553), and hospital staff without exposures (178) from Slovakia. Occupational exposures included small organic compounds (330), cytostatics (81), anesthetics (74) and metals (68). The median age was 40 years (range 19 to 71), whereas the mean was 40.6 years (s.d. 10.3); characteristics of the population are shown in Table 1. The sampling of peripheral blood was carried out according to the Helsinki Declaration and the study design was approved by the local Ethical Committee of the Jessenius Medical Faculty. The participants provided an informed consent. Cytogenetic analysis was performed on cultured lymphocytes, as previously described, by microscopically analyzing (two microscopists) in a double-blind fashion coded slides of 100 mitoses per person for the frequency of total CAs, CTAs and CSAs. Inter-arm and intra-arm interchanges of CTAs and CSAs (including dicentrics and centric rings) were also scored. In statistical analysis, non-parametric Mann–Whitney U-test was used for testing of differences between the groups. The CAs were categorized into a high-frequency group (42%) and low-frequency group (p2%); this arbitrary cutoff point was based on our previous experience. For CTA and CSA, the cutoff was 1%. Odds ratios (ORs) from multivariable linear regression analysis were employed to consider simultaneous effects of particular occupational exposures, age, gender and smoking habits on the frequencies of CAs, CTAs and CSAs. Genotyping of G870A (rs603965, also known as rs9344) was carried out by the PCR-restriction-fragment length polymorphism method (primer F 50-GTGAAGTTCATTTCCAATCCGC-30, primer R 50-GGGACATCACCCTCACTTAC-30). The amplified fragment was digested with the MspI restriction endonuclease and the digested PCR products were resolved on 3% agarose gels containing ethidium bromide for visualization under UV light. The quality control was performed by random re-genotyping of about 10% of sample. Occupational exposure information was lacking for six individuals. Genotyping of one idividual was not successful, thus 730 persons were successfully genotyped. The results for total CAs showed significant effects of occupational exposure (OR 1.68) and CCND1 AA genotype (OR 1.85, Table 1). In the separate analysis of CTAs and CSAs, the only significant effect of OR 1.99 (P1⁄4 0.003) was on CSAs (Table 2). Our recent study on multiple myeloma showed that the t(11;14) translocation was associated with the CCND1 GG genotype with an OR 1.95 (P1⁄4 2.07 10 ) (Weinhold et al.). The present results showed that AA genotype was associated with overall lymphocyte CAs and, specifically, CSAs in healthy individuals. The opposite genotype effects are likely to be explained by the cell type in question. In the earlier study, we found no genotype effect in mantle cell lymphoma, for which the t(11;14) translocation is a hallmark, most likely because in mantle cell lymphoma the translocations are formed during VDJ recombination when antigen receptor genes are rearranged, earlier than in multiple myeloma. Myeloma is thought to arise from post-germinal center memory B-cells in which immunoglobulin H (IgH) translocations take place during germinal center maturation when somatic hypermutation and class switch recombination take place. Both VDJ and class switch recombination involve complex doublestranded DNA repair events. VDJ recombination is initiated by recombination-activating genes, whereas the germinal center events are catalyzed by activation-induced cytidine deaminase, followed by classical non-homologous DNA end-joining and alternative end-joining repair activities. It is thus likely that the

Skeletal fluorosis from the point of view of an occupational exposure to fluorides in former Czechoslovakia

Skeletal fluorosis from the point of view of an occupational exposure to fluorides in former Czechoslovakia Electrolytic production of aluminium in former Czechoslovakia started in the year 1953 in the Žiar valley in the central Slovakia. However, till 1995 the hygienic conditions for health protection were not met in the factory. It underwent a reconstruction afterwards. The authors demonstrate cases of occupational skeletal fluorosis (currently rare in Europe) in 14 metallurgists which were all disclosed in foundry workers in Žiar nad Hronom as to the year 2005. The occupational disease was diagnosed after 17.7 ± 7.67 years (x ± SD) of exposure in the foundry. The authors describe the clinical conditions, haematological and biochemical tests (decreased level of ionising calcium was found in serum). The content of fluorides in urine was increased (254.4 ± 130.95 μmol/l). The average age of patients at the time of recognition of the professional etiology of the disease was 57.93 ± 7.95 years. Eight patients were older than 60 years. Skeletal abnormalities were evaluated by using X-ray skiagraphy, estimating the Stage I-III of the skeletal fluorosis. Typically an increase of bone density was found, the compact part of long bones was coarsed, there were calcifications of the interosseous membrane between radius and ulna and some ossifications of the sacrospinal and sacrotuberous ligaments. Twelve patients suffered sensorimotor polyneuropathy of extremities, chronic bronchitis was found in 6 patients (two of them were smokers). The last occupational case was registered in the year 2001. The authors assume that aluminium production with modern technology of better safety and protection of health of workers is the key which will make the skeletal fluorosis the history in the Czech and Slovak Republic.

Comparison of chromosomal aberrations frequency and polymorphism of GSTs genes in workers occupationally exposed to cytostatics or anaesthetics

Comparison of chromosomal aberrations frequency and polymorphism of GSTs genes in workers occupationally exposed to cytostatics or anaesthetics Authors compared the incidence of chromosomal aberrations (CAs) of workers occupationally exposed to cytostatics (group EXP1) or anaesthetics (group EXP2) in relationship to polymorphism of GSTM1, GSTP1 and GSTT1 genes. The cytogenetic analysis for chromosomal aberrations frequency and for polymorphisms of genes the PCR and PCR-RFLP method were used. Statistically higher frequency of total CAs was detected in both exposed groups: group EXP1 1.90±1.34%; Mann-Whitney U-test, p=0.001; group EXP2 2.53±1.46%, p=0.0008) as compared to control (1.26±0.93%). In group EXP2 was detected statistically higher frequency of aberrations CSA-type as compared to CTA-type. In xenobiotic metabolizing genes for GST higher frequency of total CAs and constituent types chromatid-type aberrations (CTAs) and chromosome-type aberrations (CSAs) of genes GSTM1 and GSTT1 with null genotype was detected. Statistically significant difference was detected only in CSA-type of aberrations in GSTT1 gene. In gene GSTP1 was not detected any difference in frequency of aberrations in presence of the variant allele. Presented results point out importance of individual susceptibility in evaluation of genotoxic agents of anaesthetics or cytostatics.

Analysis of Autonomic Nervous System Functional Age and Heart Rate Variability in Mine Workers

Abstract Introduction: Heavy working conditions and many unpropitious factors influencing workers health participate in development of various health disorders, among other autonomic cardiovascular regulation malfunction. The aim of this study is to draw a comparison of autonomic nervous system functional age and heart rate variability changes between workers with and without mining occupational exposure. Material and methods: Short term HRV was measured by DiANS PF8 device in men with and without occupational mining exposure (exposure for 10 years at least) using standard orthoclinostathic protocol (each phase 300 sec or 300 heartbeats if heart rate under 60 bpm), excluding those with severe cardiovascular, metabolic, or psychiatric diseases, nicotine and other drugs abuse, as well as those, who underwent heavy stress situation during last year. Results: Evaluating 41 HRV records among miners (n=24, age 47.9 ± 6.1 years, exposure 22.5 ± 5 years) and non-miners (n=17, age 48.5 ± 6.9 years) we found significantly increased difference between functional age of ANS and calendar age in miners group (+7.2 ± 7.3 years) over against the non-miners group (−1.6 ± 6.5 years). No correlation was found between exposure duration and functional age of ANS difference. Analysis of HRV parameters show significant reduction in total spectral power, LF, HF and rMSSD in mine workers above 48 years of age. Discussion and conclusion: Our results show that influence of occupational work factors from mining exposure can clearly take part in worsening the reactivity of ANS, which can be associated with greater risk of developing mostly cardiovascular diseases. It is important to think of non-occupational factors improving or deteriorating ANS reactivity and of individual sensitivity to other external factors.

Biomonitoring Study – Toxic Elements in Human Blood

The content of toxic heavy metals (As, Cd, Cr, Hg, and Pb) in blood depends on locality, industry and social status of habitants. We studied this determination in detail in 1994.The blood of the young healthy blood donors without any metal exposure was analyzed by the atomic absorption spectrophotometry. The results were used as the reference values of the average non-exposed population. These results did not differ from those gained in other European countries. There is no regular human biomonitoring in Slovakia but there are monitors practiced in neighbouring countries like Czech Republic or Germany. We could suppose the level of exposure in our region from these results and from literature. After 20 years the industry changed and the new technologies bring better quality of environment. It is not very appropriate to compare the results of samples which were analyzed in this time with old table values. But until this study we did not have newer data. We aimed to prepare the new bio-monitoring procedures containing recent data obtained by an inductively coupled plasma-mass spectrometry as new reference values for measurements evaluation.

Importance of GOLD Guidelines for Chronic Obstructive Pulmonary Disease

In December 2011, a major revision of GOLD 2011 guidelines was published based on the evidence-based medicine. The goal of GOLD 2011 is to determine the severity of the disease, its impact on the patients health, and the risk of future events; all of which eventually guide therapy. A combined COPD assessment according to GOLD 2011 considers the patients level of symptoms, spirometry abnormalities, risk of exacerbation, and the presence of comorbidities. GOLD 2011 stratifies patients into four basic groups labeled A, B, C, and D. The aim of the present study was to assess the importance of updated GOLD guidelines for the diagnosis, treatment, and prevention of COPD. We found that the multicomponent 2011 guidelines offer a significant advantage over the previous mono-component COPD assessment according to GOLD 2006 in terms of disease control and therapy management, with patients enjoying better spirometry values and a higher arterial oxygen content considered the primary outcomes of interest.