Variation of selected genotoxic and epigenetic markers due to therapeutic exposure to PAHs and ultraviolet radiation.

BACKGROUND: Goeckerman therapy (GT) of psoriasis involves dermal application of crude coal tar containing polycyclic aromatic hydrocarbons (PAHs) and exposure to ultraviolet radiation (UVR). Little is known about GT influence on DNA epigenetics. OBJECTIVE: The study aim was to discover epigenetic mechanisms altered by the exposure related to the GT of psoriasis. METHODS: Observed group of patients with plaque psoriasis (n = 23) was treated by GT with 3 % CCT. Before and after GT, we analyzed the levels of benzo[a]pyrene-7,8-diol-9,10-epoxide-DNA adducts (BPDE-DNA), p53 protein in serum, 5-methylcytosine (5-mC, global DNA methylation), and methylation in selected CpG sites of p53 gene. RESULTS: We found a significant increase in the levels of BPDE-DNA (p < 0.01) and serum levels of p53 protein (p < 0.01) after GT, and an insignificant decrease in the percentage of 5-mC in peripheral blood DNA. Methylation of p53 CpG sites was affected neither by psoriasis nor by GT. The study confirmed good effectiveness of GT (significantly reduced psoriasis area and severity index; p < 0.001). CONCLUSION: Our findings indicate that there is a significantly increased genotoxic hazard related to the exposure of PAHs and UV radiation after GT of psoriasis. However, global DNA methylation and p53 gene methylation evade the effect of GT, as they remained unchanged (Tab. 4, Fig. 3, Ref. 50).

Serum Levels of Aryl Hydrocarbon Receptor, Cytochromes P450 1A1 and 1B1 in Patients with Exacerbated Psoriasis Vulgaris.

The aryl hydrocarbon receptor (AhR) is highly expressed in psoriasis skin lesions. The aim of this study was to investigate serum concentrations of AhR, cytochromes P450 (CYP) 1A1 and 1B1 in patients with exacerbated psoriasis vulgaris treated with combined therapy of ultraviolet radiation (UVR) and crude coal tar. The analyses were performed by using enzyme-linked immunosorbent assays. Before the treatment, the patients had significantly higher serum levels of AhR and CYP1A1 than healthy controls. AhR median noticeably decreased after the therapy; nevertheless, it remained significantly higher compared to the controls. CYP1A1 levels measured before and after the therapy did not differ significantly. Serum CYP1A1 positively correlated with AhR values before and after the treatment. The serum values of CYP1B1 were very low and we did not see any differences between the study group and the control group. The study demonstrated that serum levels of AhR and CYP1A1 could indicate their immunopathological and metabolic roles in exacerbated psoriasis.

Elevated levels of circulating biomarkers of cell death (nucleosomes) in the patients with plaque psoriasis treated with the Goeckerman regimen.

BACKGROUND: Nucleosomes are complexes that are formed during apoptosis. Psoriasis is a chronic skin disease characterized by keratinocyte hyperproliferation and anti-apoptotic features. Presented study was focused to expression of circulating biomarkers of cell death (circulating nucleosomes, CN) during Goeckerman therapy of psoriasis (UV, PAHs). METHODS: In a group of patients with psoriasis (19), treated with Goeckerman regimen (GR), we evaluated their level of CN, level of chromosomal aberration in peripheral lymphocytes (CA), level of urinary 1-hydroxypyrene (1-OHP) and their value of Psoriasis Area and Severity Index (PASI). RESULTS: Following the treatment, the serum level of CN and urinary level of 1-OHP (p<0.05) were significantly increased (p<0.01). We found significant correlation between CN and urinary level of 1-OHP after GR (r=0.57; p<0.05). Immediately after the treatment we found significantly increased total numbers of abnormal chromosomes (ABB; p<0.01) and structurally abnormal chromosomes (SAB; p<0.05). CONCLUSIONS: We found slightly (but statistically significant) elevated level of circulating biomarkers of cell death (nucleosomes) in patients with plaque psoriasis treated with GR (PAHs, UV radiation). We suppose that elevated level of CN is a result of combination of the positive effects of GR and its weak genotoxic effect (mainly PAHs). Conclusions are supported by significant correlation between CN and urinary level of 1-OHP after GR and significantly elevated level of CA after GR (Tab. 2, Fig. 1, Ref. 28).

Expression of soluble sCD163 in serum of psoriatic patients is modulated by Goeckerman therapy.

BACKGROUND: CD163 is the monocyte/macrophage receptor for haptoglobin-haemoglobin complexes. The aim of this study was to assess the kinetics in the expression of CD163 on monocytes and the concentration of soluble sCD163 in serum of psoriatic patients in order to examine the effect of Goeckerman therapy. METHODS: sCD163 was measured in 71 patients before and after therapy, and in 57 healthy donors. A subgroup of 40 patients and 25 controls was used to assess the expression of membrane CD163. sCD163 was evaluated by ELISA. Flow cytometry method was used to determine the expression of membrane CD163 on monocytes, expressed as mean fluorescence index (MFI). RESULTS: Before therapy, the serum level of sCD163 was significantly higher in our patients than in controls (P=0.0154). However, we observed a profound decrease in sCD163 in our patients after therapy (P=0.0037). Similar to sCD163, pre-treatment expression of CD163 on monocytes was significantly more enhanced in patients than that in controls (P=0.0078). There was a trend towards down-regulation of the expression after therapy, nonetheless, the change was not statistically significant compared to the values before therapy (P=0.8666). This was also confirmed by comparison with controls which displayed lower expression of CD163 than patients after therapy (P=0.0019). The disease activity, expressed as PASI score, was significantly decreased in our patients by GT (P=0.0001). CONCLUSIONS: While sCD163 level in psoriatic patients was diminished after GT therapy, CD163 expression on monocytes was altered only to a minor extent.

C-reactive protein, chemerin, fetuin-A and osteopontin as predictors of cardiovascular risks in persons with psoriasis vulgaris.

The study aimed to contribute to understanding the role of CRP, chemerin, fetuin-A and osteopontin and to assess their suitability as biomarkers of early stages of cardiovascular diseases in psoriasis vulgaris. Serum levels measured in 28 patients and 22 controls. Patients: increased levels of CRP (p<0.001), chemerin (p<0.05), osteopontin (p<0.05) and decreased levels of fetuin-A (p<0.05), significant relationships between CRP and fetuin-A (rho=0.530, p<0.01), CRP and chemerin (rho=0.543, p<0.01), CRP and age (rho=0.590, p<0.001), osteopontin and fetuin-A (r=-0.415, p<0.05), chemerin and PASI score (rho=-0.424, p<0.05). We confirmed specific roles of the biomarkers in psoriasis. CRP, fetuin-A and osteopontin could be considered appropriate markers for the detection of early stages of cardiovascular diseases.

Plasma levels of p53 protein and chromosomal aberrations in patients with psoriasis treated with the Goeckerman regimen.

The Goeckerman regimen (GR) is one of the oldest effective treatments for psoriasis. It involves daily dermal application of crude coal tar with dermal exposure to ultraviolet (UV) radiation. A study was carried out to evaluate the genotoxic risk of GR by comparing p53 protein plasma level and chromosomal aberrations (CA) in peripheral lymphocytes in 33 patients with psoriasis, before and after GR. Psoriasis Area and Severity Index (PASI) was used to evaluate the efficacy of GR. PASI significantly decreased after GR (P < 0.001), confirming the excellent efficacy of the treatment, However, significant increases in level of p53 protein (P < 0.05) and CA (P < 0.001) after treatment indicates that this method carries an increased genotoxic risk in patients with psoriasis.

Angiogenic activity in patients with psoriasis is significantly decreased by Goeckerman's therapy.

Goeckerman's therapy (GT) of psoriasis is based on daily application of pharmacy grade coal tar on affected skin with subsequent exposure to UV light. Goeckerman's therapy is still the first line therapy of psoriasis in the Czech Republic because of its low cost and long-term efficacy. Disturbances in angiogenic activity are characteristic for the immunopathogenesis of psoriasis. An abnormal spectrum of cytokines, growth factors and proangiogenic mediators is produced by keratinocytes and inflammatory cells in patients suffering from the disease. The aim of this study was to evaluate the influence of GT of psoriasis on angiogenic activities by comparing serum levels of vascular endothelial growth factor (VEGF) and basic fibroblast growth factor (bFGF) in 44 patients with psoriasis in peripheral blood samples collected before and after therapy. Forty otherwise healthy blood donors serve as a control group. The efficacy of GT was delineated by psoriasis area and severity index (PASI). The disease activity was significantly diminished by GT (P < 0.001). The serum levels of both VEGF and bFGF were statistically significantly correlated to PASI value in patients before the treatment by GT. The serum levels of VEGF (329.4 +/- 125.5 microg/ml) and bFGF (10.2 +/- 5.04 pg/ml) in patients before GT were significantly higher than those measured in healthy blood donors (VEGF 236.4 +/- 55.9 pg/ml, bFGF 7.3 +/- 3.7 pg/ ml). The serum levels of both VEGF and bFGF were significantly diminished by GT. The level of VEGF dropped from 329.4 +/- 125.5 pg/ml before GT to 278.5 +/- 109.9 pg/ml after GT (P = 0.0042) and the level of bFGF fell from 10.2 +/- 5.04 to 7.78 +/- 4.5 pg/ml (P = 0.019). Comparing to healthy controls, the serum level of bFGF in psoriasis patients was normalised (P = 0.5723) after GT. In contrast, the serum level of VEGF remained significantly increased in psoriasis patients after GT in comparison with healthy blood donors (P = 0.0319). In conclusion, we found that the angiogenic potential which is abnormally increased in patients with psoriasis is significantly alleviated by GT.

Roles of miR-31 and endothelin-1 in psoriasis vulgaris: pathophysiological functions and potential biomarkers.

Psoriatic lesions are characterized by hyperproliferation, aberrant differentiation of keratinocytes resistant to apoptosis and inflammation. miR-31 plays pro-proliferative, pro-differentiative and pro-inflammatory roles and modulates apoptosis in psoriatic keratinocytes. Endothelin-1 (ET-1) is produced by psoriatic keratinocytes and suppresses apoptosis. Inflammation increases the production of ET-1, which in turn leads to the chronic stimulation of keratinocyte proliferation. The aim of this study was to identify the putative link between two potential biomarkers (miR-31 and ET-1) in patients with psoriasis. The study design included experimental group (29 patients with psoriasis), and the control group (22 blood donors). The PASI score evaluated the state of the disease (median: 18.6; interquartile range 14.5-20.9). Both, the serum level of ET-1 and the whole blood level of miR-31 were significantly increased (p<0.001 and p<0.05, respectively) in patients compared to the controls. However, a significant negative relationship between ET-1 and miR-31 was observed (Spearman's rho=-037, p=0.05). It is possible that a negative feedback loop will be present between miR-31 and ET-1. Our results indicate that miR-31 and ET-1, potential biomarkers of the disease, play significant roles in the pathophysiology of psoriasis.

Selected immunological changes in patients with Goeckerman's therapy TNF-alpha, sE-selectin, sP-selectin, sICAM-1 and IL-8.

Psoriasis is one of the most frequent inflammatory skin diseases in which abnormal individual immune reactivity plays an important role. The aim of the present study was to describe selected immunological changes, concerning pro-inflammatory cytokines (TNF-alpha, IL-8) and adhesion molecules (sE-selectin, sP-selectin, sICAM-1), in 56 patients cured by Goeckerman's therapy (GT). GT includes dermal application of crude coal tar (containing polycyclic aromatic hydrocarbons) and exposure to UV radiation. When compared with the control group (healthy blood donors), the patients before GT had significantly increased serum levels of sE-selectin (p<0.001), sP-selectin (p<0.001), sICAM-1 (p<0.001) and IL-8 (p<0.001). Significantly decreased serum levels of sE-selectin (p<0.05) and significantly increased serum levels of IL-8 (p<0.05) were found after GT therapy. Serum levels of sICAM significantly correlated with the disease activity and with serum levels of sE-selectin. The level of PASI score (Psoriasis Area and Severity Index) significantly decreased after GT (p<0.001) and confirms the high efficiency GT. These findings confirmed that pro-inflammatory chemokine (IL-8) and adhesion molecules (sE-selectin, sP-selectin, sICAM-1) play an important role in the development and regulation of inflammation in psoriasis. Determination of sE-selectin and sICAM seems to be a promising marker of psoriasis's activity. Chemokine pathway (IL-8) and TNF-alpha activity seem to be modulated by Goeckerman's therapy (polycyclic aromatic hydrocarbons).

Cytogenetic and immunological changes after dermal exposure to polycyclic aromatic hydrocarbons and UV radiation.

Goeckerman's therapy (GT), which combines exposure to coal tar (polycyclic aromatic hydrocarbons - PAHs) and UV radiation (UV) is often used as the first option for treatment of psoriasis. However, PAHs and UV represent mutagenic, carcinogenic and immunotoxic agents. Therefore GT can represent a health risk for the patients. The group under observation consisted of thirty patients undergoing GT. Before and after the treatment, blood samples were collected and chromosomal aberrations and selected immunological markers were determined. The relationships between chromosomal aberrations and immunological markers and the extent (duration) of exposure to GT were evaluated. The Psoriasis Area and Severity Index (PASI) score confirmed the high efficacy of GT. However, significantly elevated levels of chromosomal aberrations of peripheral lymphocytes were also found after the therapy (p<0.001). The levels of chromosomal abnormalities correlated to the extent and the total duration of exposure to PAHs (r = 0.682, p<0.01 and r = 0.605, p<0.05). After the therapy, significantly decreased levels of IgE, IgM isotypes of immunoglobulin, alpha(2)-macroglobulin and transferrin together with beta(2)-microglobulin were found. From the immunological markers listed above only the decreased level of alpha(2)-macroglobulin correlated to the extent of exposure to PAHs (r = -0.568, p<0.05). No correlation was found between chromosomal aberrations, significantly changed immunological markers and the duration of UV exposure. Our study revealed that GT has a significant impact on both genetic and immunological parameters of psoriatic patients. The results indicate that GT could increase genotoxic risk and modulates immunity of treated patients.

Distribution of nickel in body fluids and organs of rats chronically exposed to nickel sulphate.

1. Male and female rats were given 100 mg Ni L-1 (as nickel sulphate) in drinking water for 6 months. 2. The feeding of nickel was associated with an increased concentration of nickel in body fluids and organs. The highest concentrations of nickel were found in the liver of both male and female rats. In male rats nickel levels decreased in the order: liver > kidney = whole blood = serum > testes > urine. In female rats the decreasing order was similar: liver > kidney = whole blood = serum = plasma > urine > ovaries. 3. No significant differences were found between nickel concentrations in organs (except ovaries), blood and urine of rats exposed for 3 months and those exposed for 6 months indicating the reaching of a steady state of nickel in the rat during long-term exposure. 4. The urinary excretion of the orally administered nickel was only 2% of absorbed dose (supposing 1% Ni absorption).

Environmental exposure to polycyclic aromatic hydrocarbons in Czech Republic.

1 Objectives of this study were (1) to compare concentrations of individual polycyclic aromatic hydrocarbons (PAH) in air of polluted and nonpolluted area of Czech Republic during winter and summer periods and (2) to verify if urinary 1-hydroxypyrene (1-OHP), as supposed practical biological marker, permits the assessment of spacial and temporal variations in environmental PAH exposure. 2 The study population consisted of three groups: (1) a group of 22 physical exercise students who regularly train outside, from the university situated in a polluted town, spending 14 days in winter and 14 days in summer in 'non-polluted' mountains; (2) a control group of 22 residents from the town and (3) a control group of 18 residents from the mountains. 3 The total PAH concentrations (sum of 13 individual PAH) were 19.3 and 104.6 ng/m3 in town and in mountains, respectively, during summer and 86.6 and 261 ng/m3 during winter. 4 Median 1-OHP levels ranged between 0.03 and 0.13 mumol/mol creatinine for controls and between 0.04 and 0.12 mumol/mol creatinine for students. No relationship was found between pyrene levels in air and group means of urinary 1-OHP. Our results show that other factors (probably PAH in food) contribute in masking air pollution influence on urinary 1-OHP levels in subjects non-occupationally exposed to PAH.

Study on kidney function in female workers exposed to perchlorethylene.

1. Biochemical markers of kidney damage were examined in 16 female workers chronically exposed to tetrachlorethylene (TCE) in five dry-cleaning shops. The results were compared with those obtained in 13 females non-occupationally exposed to organic solvents. 2. The intensity of exposure was monitored by personal environmental monitoring. The time-weighed average exposure to TCE amounted to 157 mg m-3 (range 9-799 mg m-3). A satisfactory agreement was found between the concentration of TCE in ambient air sampled with the charcoal tube method and with a passive dosimeter. 3. The urinary excretion of lysozyme was increased in the exposed group. No difference was found in the urinary excretion of albumin, beta 2-microglobulin, lactate dehydrogenase, total proteins or glucose. The prevalence of abnormal values of biochemical parameters in the exposed group did not differ from that observed in the control group. No correlation was found between the level of TCE exposure and biochemical parameters. 4. The present study suggests that chronic exposure to TCE does not lead to renal damage.

Environmental health.

A short history of the close connections between human health an ambient environment (environmental health) in the global and national (Czech Republic) sense is presented. Concrete tasks and prospects for the near future are mentioned, including the human ecology education programme.

Possible genotoxic risk of combined exposure to pharmaceutical coal tar and UV-B radiation.

Goeckerman's therapy of psoriasis combines exposure to pharmaceutical coal tar and UV-B radiation. In the pilot study (15 patients had been diagnosed with psoriasis, the average time period in hospital therapy was 24 days, the average age of the patients was 29 years, 47% of them were smokers) a level of genotoxic risk from therapy was evaluated by using chromosomal aberration of peripheral lymphocytes. The study suggested the presence of an increased genotoxic risk from the therapy. The PASI scores (Psoriasis Area and Severity Index) were monitored.

The effects of subchronical exposure to SO2 on biochemical and hematological parameters in guinea pigs.

The effects of subchronical exposure to SO2 (400ppm, 3 hours daily, 28 days) on biochemical and hematological parameters were investigated in guinea pigs. Mostly no significant changes in the values of biochemical parameters and no significant changes in hematological parameters were found. The levels of investigated ions (K+, Na+, Cl-, Ca++, Mg++ and phosphates), proteins (albumines, globulines, total proteins), enzymes (LD, ALT, AST, CK) and other biochemical parameters (urea, creatinine, bilirubin) were not significantly different between groups, with the exception of a significantly higher ALP concentration in the exposed group as compared with controls (2.17 mukat and 1.85 mukat, respectively. It can be concluded that a subchronical exposure to sulphur dioxide mostly did not induce any definite changes in biochemical and hematological parameters in guinea pigs.

[Biological monitoring of chemical exposure]. / Biologické monitorování chemické expozice.

In this paper we described the biological monitoring as a capable exposure assessment tool that has provided important information used in public health decisions. Biological monitoring is based on determination of biological markers of exposure which are presented as the quantity of a chemical substance or its metabolites or as the deviation of biological parameters (enzyme activity etc.) induced by this substance in exposed humans. The greatest advantage of biological monitoring is the fact that the biological marker of exposure is more directly related to the adverse effects than any environment measurement. Another advantage of biological monitoring is based on the reality that the nonoccupational background exposure (leisure activity, residency, dietary habits, smoking, etc.) may also be expressed in the biological level. Biological parameters can be unfortunately affected by various factors that influence the fate of xenobiotic in vivo. The "BEL" (BTV-biological tolerance value for occupational exposures) is defined as the maximum permissible quantity of a chemical substance or its metabolites or the maximum permissible deviation from the norm of biological parameters during or after exposure. It should be subject to regular revision in the light of new scientific data.

[Phenotyping of cytochrome P-450 1A2 and N-acetyltransferase (NAT2) using the in vivo caffeine test as a tool for determining individual susceptibility to selected xenobiotics]. / Fenotypizace cytochromu P-450 1A2 a N-acetyltransferasy (NAT2) in vivo metodou kofeinového testu jako nástroj pro zjistování individuální vnímavosti k vybraným xenobiotikum.

An in vivo phenotyping method of CYP1A2 (cytochrome P-450 1A2) and NAT2 (N-acetyltransferase, isoform 2) based on caffeine test (Butler et al. Pharmacogenetics 1992;2:116-127) with modifications has been established. Molar concentrations of caffeine and its metabolites in urine were determined by HPLC with gradient elution and spectral detection (200-350 nm). The results obtained with this method were sufficiently accurate and precise and allowed to calculate the respective parameters which were subsequently used to evaluate the CYP1A2 and NAT2 activities.

[Effect of acute exposure to SO2 on cellular defense mechanisms in the lung (pilot study)]. / Ucinek akutní expozice SO2 na celulární obranné mechanismy plic (pilotní studie).

Sulphur dioxide is one of the most important and most frequent atmospheric pollutant of our environmental setting. In presented pilot-study we describe the influence of acute exposure to sulphur dioxide on alveolar macrophages of guinea pigs. The alveolar macrophages represent the main part of mononuclear phagocytic system, which is responsible for the defence of respiratory tract against foreign compounds. The experimental animals were exposed over three hours to sulphur dioxide with the concentration of about 400 ppm. After exposure we did not find significant changes in phagocytic activity of alveolar macrophages.

[Chemical load in the population and its evaluation]. / Chemická zátez populace a její hodnocení.

Human health is determined by the interplay between heredity and the environment. Air, water, food and soil contain chemical, physical and biological agents some of which are known to be harmful to health. Chemical substances that pose the risk to human health and safety and to the environment are subject to governmental regulation. The regulatory decision-making process and regulatory actions are based on two distinct elements: risk assessment and risk management. Air pollution (outdoor, indoor) is a world problem afflicting densely populated urban centers and heavily industrialised areas. Industrialization and the widespread use of chemicals coupled with modern intensive agricultural practices have raised a global concern about the contamination of soil and water. Three categories of environmental chemical contaminants generally occur in food--natural and synthetic organic compounds and traces of toxic metals. Human health protection against chemical exposure can be realised in three ways. Environmental monitoring assesses exposure to a chemical agent by measuring its concentration in the environment (i.e., air, soil, food, water). Biological monitoring assesses internal exposure to a chemical agent by measuring the chemical, its metabolites or nonadverse biological response in body fluids, tissues, expired air or excreta. Health surveillance entails the periodic medical examinations of exposed humans with the purpose of protecting health and preventing disease.