Wojciech Wąsowicz

Publications of Wojciech Wąsowicz

• Evaluation of Reproductive Disorders in Female Rats Exposed to N-Methyl-2-Pyrrolidone.

  Authors: Krystyna Sitarek, Jan Stetkiewicz, Wojciech Wąsowicz

  Birth defects research. Part B, Developmental and reproductive toxicology. 02/2012;

  BACKGROUND: N-methyl-2-pyrrolidone (NMP) is a solvent used in the petrochemical, and electronic industries, in pesticides production, veterinary drugs, and paint removers. The aim of study was toBACKGROUND: N-methyl-2-pyrrolidone (NMP) is a solvent used in the petrochemical, and electronic industries, in pesticides production, veterinary drugs, and paint removers. The aim of study was to evaluate the relationship between the dose of NMP given orally and its effect on fertility in female rats and early development of their progeny. METHODS: Females were exposed by gavage 5 days/week to NMP at 150, 450, or 1000 mg/kg/day 2 weeks before mating, during mating, gestation, and lactation. On the first postnatal day (PND 1), the live and dead pups were counted, weighed, and gender was determined. On PND 4, the litters were culled to eight animals each and balanced for gender. Young animals were observed during 3 weeks after birth. RESULTS AND CONCLUSION: Fertility index did not significantly differ in the control and the group exposed at 150 mg/kg/day but it was significantly lower in the groups exposed at 450 or 1000 mg/kg/day. The number of live pups in the group exposed to the highest dose was significantly lower and the number of stillbirths in litters was significantly greater. Survival of the pups from all exposed groups during the 3 weeks after birth was significantly lower than the control animals. The results of our study indicate that intragastric exposure of female rats to NMP before pregnancy during gestation causes significant impairment in female fertility and intrauterine mortality rates. At lower doses, toxic or slightly toxic to the mothers, this substance causes decrease in viability and physical development of progeny.

• Evaluation of biological effects of nanomaterials. Part I. Cyto- and genotoxicity of nanosilver composites applied in textile technologies.

  Authors: Wojciech Wąsowicz, Małgorzata Cieślak, Jadwiga Palus, Małgorzata Stańczyk, Elżbieta Dziubałtowska, Maciej Stępnik, Markus Düchler

  International journal of occupational medicine and environmental health. 12/2011; 24(4):348-58.

  The aim of this study was to investigate the cyto- and genotoxicity of nanocomposites (NCs) and generation of reactive oxygen species (ROS) as a result of particle-cell interactions. Titanium dioxideThe aim of this study was to investigate the cyto- and genotoxicity of nanocomposites (NCs) and generation of reactive oxygen species (ROS) as a result of particle-cell interactions. Titanium dioxide (TiO(2)-Ag) and ion-exchange resin (Res-Ag), both coated with silver (Ag), were examined. The murine macrophage J774A.1 cells were incubated in vitro with NC at different concentrations for 24 h. Cytotoxicity was analyzed by the methylthiazolyldiphenyl-tetrazolium bromide reduction test (MTT reduction test). ROS generation was assessed by incubation of cells with dichlorodihydrofluorescein diacetate (DCF) and flow cytometry. DNA damage was detected by comet assay and included single-strand breaks (SSB), alkali-labile sites (ALS) and oxidative DNA damage after formamidopyrimidine glycosylase (FPG) treatment. The tail moment was used as an indicator of DNA damage. TiO(2)-Ag was not cytotoxic up to 200 μg/ml, whereas IC(50) for Res-Ag was found to be 23 μg/ml. Intracellular ROS levels were elevated after 4 h of exposure to Res-Ag at the concentration of 50 μg/ml. Both types of NC induced fragmentation of DNA strands, but only one of the composites caused damage to purine bases. TiO(2)-Ag induced SSB of DNA at concentrations of 10 and 5 μg/ml. For Res-Ag, a concentration-dependent increase in tail moments was observed. Silver-coated nanocomposites (both TiO(2)-Ag and Res-Ag) may cause genotoxic effects in murine macrophages J774A.1. Res-Ag increased generation of ROS which suggested that toxicity of Res-Ag in murine macrophages is likely to be mediated through oxidative stress. This paper will support industry and regulators alike in the assessment of hazards and risks and methods for their mitigation at the earliest possible stage in material and product development.

• 4-Week inhalation toxicity of 2-methylnaphthalene in experimental animals.

  Authors: Radosław Swiercz, Wojciech Wąsowicz, Jan Stetkiewicz, Jolanta Gromadzińska, Wanda Majcherek

  International journal of occupational medicine and environmental health. 12/2011; 24(4):399-408.

  This paper presents toxic effects of 2-MN in laboratory animals under conditions of 4-week inhalation exposure to 2-methylnaphthalene (2-MN) vapors. Male Wistar rats were exposed to 2-MN vapors at aThis paper presents toxic effects of 2-MN in laboratory animals under conditions of 4-week inhalation exposure to 2-methylnaphthalene (2-MN) vapors. Male Wistar rats were exposed to 2-MN vapors at a nominal concentration of 0, 2, 10 or 50 mg/m(3) in dynamic inhalation chambers for 4 weeks (6 h/day, 5 days/week). After 4 weeks of inhalation exposure the animals were necropsied. Blood samples were collected and selected organs were weighted and prepared for histological examinations. The effects of the increased levels of exposure to 2-MN experienced by the experimental rats were as follows: a) increasing γ-glutamylotransferase activity, b) stimulation of the hematopoietic system, c) lower cholesterol concentrations, d) higher number of goblet cells in lobar bronchi, e) hyperplasia of hepatic bile ducts. Four-week exposure of the animals to 2-MN at 2 mg/m(3) proved to be the no-observed-adverse-effect-level (NOAEL), while 10 mg/m(3) appeared to represent the lowest-observed-adverse-effect-level (LOAEL).

• Oxidative DNA damage and oxidative stress in subjects occupationally exposed to nitrous oxide (N(2)O).

  Authors: Teresa Wrońska-Nofer, Jerzy-Roch Nofer, Jolanta Jajte, Elżbieta Dziubałtowska, Wiesław Szymczak, Wojciech Krajewski, Wojciech Wąsowicz, Konrad Rydzyński

  Mutation research. 11/2011; 731(1-2):58-63.

  Occupational exposure to nitrous oxide (N(2)O) and/or halogenated hydrocarbons has been suggested to induce damage of genetic material, but the underlying mechanisms remain obscure. This studyOccupational exposure to nitrous oxide (N(2)O) and/or halogenated hydrocarbons has been suggested to induce damage of genetic material, but the underlying mechanisms remain obscure. This study investigated the role of oxidative processes in the genotoxicity associated with exposure to waste anaesthetic gases. The study was performed in 36 female nurses and in 36 unexposed female health care workers matched for age and employment duration. Genotoxic effects were examined by Comet test modification employing formamidopyrimidine glycosylase (FPG) that allows assessment of oxidative DNA damage. Reactive oxygen species (ROS) in leukocytes were investigated by fluorescence spectroscopy with 2',7'-dichlorofluorescin diacetate. Oxidative stress markers including 8-iso-prostaglandin F(2α) (8-iso-PGF(2α)), thiobarbituric acid-reacive substances (TBARS), α-tocopherol, and glutathione peroxidise (GPX) activity were measured immuno- or colorimetrically. N(2)O, sevoflurane and isoflurane were monitored by gas chromatography and mass spectrometry. The study documents for the first time the positive correlation between the oxidative DNA damage and the N(2)O levels in the ambient air. By contrast, no association was observed between genotoxic effects and sevoflurane or isoflurane. In addition, ROS generation and plasma and urine concentrations of TBARS and 8-iso-PGF(2α), respectively, were elevated, while GPX activity was reduced in nurses exposed to waste anaesthetic gases. Path analysis pointed to a causal relationship between N(2)O exposure, oxidative stress and DNA damage. Occupational exposure to N(2)O is associated with increased oxidative DNA damage and the level of exposure plays a critical role in this regard. Increased oxidative stress may represent a mechanistic link between chronic N(2)O exposure and genotoxicity.

• GSTP1 mRNA expression in human circulating blood leukocytes is associated with GSTP1 genetic polymorphism.

  Authors: Edyta Reszka, Zbigniew Jabłonowski, Edyta Wieczorek, Jolanta Gromadzińska, Marek Sosnowski, Wojciech Wąsowicz

  Clinical biochemistry. 06/2011; 44(13):1153-5.

  We explored association between GSTP1 Ile(105)Val (rs1695) polymorphism and GSTP1 mRNA expression in circulating blood leukocytes. GSTP1 transcripts level and polymorphism were determined byWe explored association between GSTP1 Ile(105)Val (rs1695) polymorphism and GSTP1 mRNA expression in circulating blood leukocytes. GSTP1 transcripts level and polymorphism were determined by Real-Time PCR in 51 bladder cancer and 90 healthy men. Individuals with at least one GSTP1 Val(105) variant allele possessed higher GSTP1 mRNA level in blood leukocytes compared to GSTP1 Ile(105)Ile carriers. GSTP1 Ile(105)Val gene polymorphism influences its expression in blood, regardless of cancer disease.

• Hypermethylation of p16 and DAPK promoter gene regions in patients with non-invasive urinary bladder cancer.

  Authors: Zbigniew Jabłonowski, Edyta Reszka, Jolanta Gromadzińska, Wojciech Wąsowicz, Marek Sosnowski

  Archives of medical science : AMS. 06/2011; 7(3):512-6.

  The aim of the study was to examine the frequency of methylation status in promoter regions of p16 and DAPK genes in patients with non-invasive bladder cancer. Forty-two patients (92.9% men, 73.8%The aim of the study was to examine the frequency of methylation status in promoter regions of p16 and DAPK genes in patients with non-invasive bladder cancer. Forty-two patients (92.9% men, 73.8% smokers, 71.4% T1G1, 19.1% T1G2, 9.5% T1G3) and 36 healthy controls were studied. Isolation of genomic DNA from blood serum and methylation-specific PCR (MSP) were applied. Methylation status - methylated and unmethylated promoter regions of p16 and DAPK genes were analysed. Seventeen out of 42 patients (40.5%) had the methylated p16 gene, while methylation of the DAPK gene was seen in 27 of 42 cases (64.3%). In 12 patients (28.6%) both analysed genes were methylated. A statistically significant (p = 0.046) higher frequency of DAPK gene methylation (71.4%) was observed in patients with lower grade (G1) bladder cancer. Detection of the aberrant hypermethylation of DAPK and p16 genes in blood DNA from non-invasive bladder cancer patients might offer an effective means for earlier auxiliary diagnosis of the malignancy.

• The toxicokinetics of 2-methylnaphtalene in rats.

  Authors: Radosław Swiercz, Wojciech Wąsowicz, Wanda Majcherek

  International journal of occupational medicine and environmental health. 01/2010; 23(4):385-9.

  The aim of the study was to evaluate the toxicokinetics of 2-methylnaphtalene (2-MN) during and after inhalation exposure. Male Wistar rats were exposed to 2-MN vapours at nominal concentrations ofThe aim of the study was to evaluate the toxicokinetics of 2-methylnaphtalene (2-MN) during and after inhalation exposure. Male Wistar rats were exposed to 2-MN vapours at nominal concentrations of 200 or 400 mg/m3 in the dynamic inhalation chamber for 6 hours or 5 days (6 h/day). Blood samples were collected during and after exposure. Blood concentrations of 2-MN were estimated by gas chromatography using the headspace technique. During a 6-hour exposure to 200 or 400 mg/m3, blood 2-MN concentration increased rapidly within the first or second hour of exposure, respectively, after reaching a plateau. The elimination of 2-MN from blood followed an open two-compartment model. 2-MN was rapidly eliminated from blood of the animals exposed by inhalation to 2-MN. During exposure, lung retention of the chemical was found to decrease. Under conditions of repeated 2-MN exposure, no significant systemic 2-MN accumulation could be observed.

• Catecholamine levels in the brain of rats exposed by inhalation to benzalkonium chloride.

  Authors: Radosław Swiercz, Zofia Grzelińska, Sławomir Gralewicz, Wojciech Wąsowicz

  International journal of occupational medicine and environmental health. 07/2009;

  Objectives: The aim of the study was to obtain quantitative data on the effect of inhalation exposure to benzalkonium chloride (BAC) on the concentration of catecholamines and their metabolites inObjectives: The aim of the study was to obtain quantitative data on the effect of inhalation exposure to benzalkonium chloride (BAC) on the concentration of catecholamines and their metabolites in selected brain structures. Additionally, concentration of corticosterone (CORT) in plasma was estimated. Material and Methods: Wistar rats were subjected to a single (6-hour) or repeated (3 days, 6 h/day) exposure to BAC aerosol at ca. 30 mg/m3. The Waters integrated analytical system of HPLC was used to determine the plasma corticosterone. Qualitative and quantitative determinations of catecholamines and their metabolites: 3,4-dihydroxyphenylacetic (DOPAC) and homovanillic (HVA) acids were performed with the use of the Waters integrity HPLC. Results: The determinations have shown that in the BAC-exposed rats the plasma CORT concentration was several times higher than in the control rats. A significant increase of the concentration of dopamine (DA) (striatum and diencephalon) and noradrenaline (NA) (hippocampus and cerebellum) and a significant reduction of adrenaline (A) level (cortex, hippocampus, striatum and mesencephaloon) was found to occur in the brain of rats exposed to BAC compared to control. In the animals exposed to BAC, the concentration of DOPAC, a DA metabolite, was significantly reduced, but the change occurred mainly in the striatum. This resulted in a significant decrease of the DOPAC/DA and HVA/DA metabolic ratio in this structure. Conclusion: It is assumed that the alterations in the concentration of catecholamines and their metabolites in the BAC-exposed rats were related to the unexpectedly strong and persistent activation of the hypothalamo-pituitary-adrenocortical (HPA) axis evidenced by the high plasma CORT concentration.

• DNA damage induced by nitrous oxide: Study in medical personnel of operating rooms.

  Authors: Teresa Wrońska-Nofer, Jadwiga Palus, Wojciech Krajewski, Jolanta Jajte, Małgorzata Kucharska, Jan Stetkiewicz, Wojciech Wąsowicz, Konrad Rydzyński

  Mutation research. 05/2009;

  Occupational exposure to anaesthetics such as nitrous oxide (N(2)O) and halogenated hydrocarbons has been suggested to increase risk of genetic damage. However, the dose-dependency of genotoxicOccupational exposure to anaesthetics such as nitrous oxide (N(2)O) and halogenated hydrocarbons has been suggested to increase risk of genetic damage. However, the dose-dependency of genotoxic effects has not been unequivocally established and their relation to occupational exposure limit (OEL) remain obscure. In this study, the genotoxicity associated with occupational exposure to anaesthetics has been investigated in a group of 55 female nurses and 29 male anaesthesiologists active for at least 5 years in a working environment containing variable concentrations of N(2)O and halogenated hydrocarbons. 83 unexposed health care workers (52 female nurses and 31 male doctors) matched for age, gender, smoking habit and employment duration were included in the control group. Genotoxicity has been assessed using comet test. Concentrations of nitrous oxide, sevoflurane and isoflurane monitored by gas chromatography and mass spectrometry made possible to relate the extent of DNA damage to the level of exposure. Our results for the first time document a positive correlation between the DNA damage and the N(2)O levels in the ambient air. By contrast, no correlation has been observed between genotoxic effects and concentrations of sevoflurane and isoflurane. The extent of genetic injury was especially aggravated among nurses and anaesthesiologists exposed to N(2)O in concentrations exceeding OEL (180mg/m(3)). We conclude that occupational exposure to N(2)O is associated with increased DNA damage and that the level of exposure plays a critical role in this regard.

• Carcinogenic effect of arsenate in C57BL/6J/Han mice and its modulation by different dietary selenium status

  Authors: Maciej Stępnik, Jan Stetkiewicz, Aleksander Krajnow, Katarzyna Domeradzka, Dobrosława Gradecka-Meesters, Joanna Arkusz, Małgorzata Stańczyk, Jadwiga Palus, Elżbieta Dziubałtowska, Wojciech Sobala, Jolanta Gromadzińska, Wojciech Wąsowicz, Konrad Rydzyński

  Ecotoxicology and Environmental Safety.

  In this study, carcinogenic effects of arsenate in female C57BL/6J/Han mice exposed in drinking water to 50, 200 or 500 μg As/L for 24 months were investigated. All animals were fed low-seleniumIn this study, carcinogenic effects of arsenate in female C57BL/6J/Han mice exposed in drinking water to 50, 200 or 500 μg As/L for 24 months were investigated. All animals were fed low-selenium diet, however half of them were supplemented with sodium selenite in drinking water (200 μg Se/L) to ensure the normal dietary level of selenium. Glutathione peroxidase activity in erythrocytes and plasma as well as selenium concentration in plasma after 3, 6, 12 and 18 months in satellite groups showed considerable decrease in animals from non-selenium supplemented groups in comparison to supplemented groups. A clear arsenic concentration-dependent increase in the number of malignant lymphoma associated with increase in the risk of death was observed (hazard ratio=0.91, 1.46, and 2.24, for 50, 200 and 500 μg As/L, respectively). No significant influence of selenium dietary status on arsenic carcinogenicity was shown. A significant association between selenium supplementation status and increased risk of death of the animals from causes other than malignant tumors was found (HR=1.79, p=0.04).