Concentration of trace elements in blood of Polish patients with prostate cancer.

The goal of the study was to analyse the concentrations of chemical elements (Fe, Ni, As, Cd, Pb, Hg, Cr, Zn) which are important for the determination of environmental toxins (e.g. resulting from smoking, exposure to harmful agents at work) in Polish patients with prostate cancer. The study covered 66 patients with diagnosed prostate cancer and 64 healthy volunteers over 50 years old. The analysis of the concentrations of selected chemical elements in whole blood was performed using inductively coupled plasma mass spectrometry (ICP-MS). In their blood, the patients with cancer had a significantly higher concentration of only one of the examined elements: arsenic. Additionally, the study group had lower concentrations of chromium, zinc, but also cadmium and lead, which are commonly regarded as carcinogenic. Taking into consideration the control group of healthy subjects of this study, we can assume that the subjects with prostate cancer were exposed to higher levels of arsenic, and that exposure to this element may be associated with an increased risk of cancer.

The Reaction of the Yeast Saccharomyces cerevisiae to Contamination of the Medium with Aflatoxins B2 and G1, Ochratoxin A and Zearalenone in Aerobic Cultures.

The mechanisms by which yeast cells respond to environmental stress include the production of heat shock proteins (HSPs) and the reduction of oxidative stress. The response of yeast exposed to aflatoxins B2+G1 (AFB2+G1), ochratoxin A (OTA), and zearalenone (ZEA) in aerobic conditions was studied. After 72 h of yeast cultivation in media contaminated with mycotoxins, the growth of yeast biomass, the level of malondialdehyde, and the activity of superoxide dismutase, glutathione S-transferase and glutathione peroxidase were examined; the expression profile of the following heat shock proteins was also determined: HSP31, HSP40, HSP60, HSP70, and HSP104. It was demonstrated that at the tested concentrations, both AFB2+G1 and ZEA inhibited yeast biomass growth. OTA at a concentration of 8.4 [µg/L] raised the MDA level. Intensified lipoperoxidation and increased activity of SOD and GPx were observed, regardless of the level of contamination with ZEA (300 µg/L or 900 µg/L). Increased contamination with AFB2+G1 and OTA caused an increase in the production of most HSPs tested (HSP31, HSP40, HSP70, HSP104). ZEA contamination in the used concentration ranges reduced the production of HSP31. The response of yeast cells to the presence of mycotoxin as a stressor resulted in the expression of certain HSPs, but the response was not systematic, which was manifested in different profiles of protein expression depending on the mycotoxin used. The tested mycotoxins influenced the induction of oxidative stress in yeast cells to varying degrees, which resulted in the activation of mainly SOD without GST mobilization or with a small involvement of GPx.

Analysis of oxidative stress indicators in Polish patients with prostate cancer.

The aim of the study was to analyze the activity of antioxidant enzymes (glutathione S-transferase, catalase, superoxide dismutase) and the concentration of malondialdehyde in order to determine the role of detoxification mechanisms in prostate cancer. The activities of superoxide dismutase (SOD), catalase (CAT), and glutathione S-transferase (GST) were measured using ready-made kits; lipid peroxidation intensity was determined by the thiobarbituric acid method. Superoxide dismutase was the only enzyme among antioxidant and detoxification enzymes for which a statistically significant difference in activity was found between the studied groups (1.4 U·ml-1 in patients vs. 1.6 U·ml-1 in control). No statistically significant differences were found for GST, CAT or the concentration of MDA between the group of men with prostate cancer and the control group. The lower SOD activity in men with prostate cancer may be due to a deficiency in their antioxidant defense system.

Influence of the Medium Composition and the Culture Conditions on Surfactin Biosynthesis by a Native Bacillus subtilis natto BS19 Strain.

An effective microbial synthesis of surfactin depends on the composition of the culture medium, the culture conditions and the genetic potential of the producer strain. The aim of this study was to evaluate the suitability of various medium components for the surfactin producing strain and to determine the impact of the culture conditions on the biosynthesis of surfactin isoforms by the newly isolated native strain Bacillus subtilis natto BS19. The efficiency of surfactin biosynthesis was determined by measuring the surface tension of the medium before and after submerged culture (SmF) and by qualitative and quantitative analysis of the obtained compound by high performance liquid chromatography. The highest efficiency of surfactin biosynthesis was achieved using starch as the carbon source and yeast extract as the nitrogen source at pH 7.0 and 37 °C. Potato peelings were selected as an effective waste substrate. It was shown that the increase in the percentage of peel extract in the culture medium enhanced the biosynthesis of surfactin (mg/L) (2-30.9%; 4-46.0% and 6-58.2%), while reducing surface tension of the medium by about 50%. The obtained results constitute a promising basis for further research on biosynthesis of surfactin using potato peelings as a cheap alternative to synthetic medium components.

Antioxidant defence barrier of great tit Parus major nestlings in response to trace elements.

Metals can have direct and indirect effects on the generation of reactive oxygen species in wild birds. The aim of this work has been to examine the effect of exposure to trace metals (copper Cu, iron Fe, cobalt Co, manganese Mn) on oxidative stress biomarkers such as lipoperoxidation TBARS and level of superoxide dismutase SOD, catalase CAT, and reduced glutathione GSH in the livers and kidneys of great tit Parus major nestlings (n = 165, 63 broods) living in polluted environments associated with soda plants and agricultural activities (Kujawy region) and from a reference site (Tuchola Forest), both in the north of Poland. As we predicted, the level of TBARS in both organs of chicks from polluted areas was higher than in those from reference site. This could be connected with Fe concentrations, particularly in areas adjacent to soda plants (livers Rs = 0.49, p < 0.002; kidneys Rs = 0.69, p < 0.001). We also showed differences in the level of antioxidants depending on the environment. CAT activity was higher in nestlings from Kujawy than in those from Tuchola. Meanwhile SOD activity (both organs) and GSH levels (kidneys) were lower in the polluted area compared to the reference site. Concentrations of Cu, Fe, Co, and Mn may play a role in regulating the antioxidant system components' activity.

Polymorphism of glutathione S-transferase in the population of Polish patients with carcinoma of the prostate.

The aim of the study was frequency analysis of GSTM1, GSTT1, and GSTP1 polymorphisms of glutathione S-transferase in the group of patients with prostate cancer and in a control group of healthy individuals. Genomic DNA was isolated; molecular analysis of glutathione S-transferase M1 and T2 polymorphisms was performed using multiplex PCR and RFLP methods. The products of the PCR reaction were then visualized in agarose gel, and a statistical analysis of the results was performed. No statistically significant differences were found in the frequency of glutathione S-transferase polymorphisms between 66 patients with prostate cancer and the control group (64 healthy volunteers). The GSTM1 gene deletion was found in ca. 47% of patients with prostate cancer and in ca. 55% of the controls. The GSTT1 deletion was found in approximately 17% of patients and 14% of the controls. The distribution of GSTP1 Ile/Ile, Ile/Val, and Val/Val polymorphisms was ca. 51.5%, 39%, and 9% in the group of patients and 61%, 34%, and 5% in the control group, respectively. The results indicate that there is no relationship between glutathione S-transferase polymorphisms and prostate cancer in the study group, which is a novelty when compared with the previous work on the role of these genetic variants in the etiology of cancer.

Evaluation of various methods of selection of B. subtilis strains capable of secreting surface-active compounds.

The aim of the study was the evaluation of a three-step method for the selection of bacterial strains capable of producing surfactin. The procedure consisted of the following steps: 1.blood agar test, 2. measurement of the surface tension (ST) of the medium using the du Nouy method before and after submerged culture, 3. qualitative and quantitative assessment of surfactin by HPLC. Forty five Bacillus subtilis natto strains producing haemolysis zones (≥3mm) were selected. Nineten of them reduced ST of the medium to ≤ 40 mN/m; in six cases, the reduction was as much as 50%. All indicated strains produced surfactin. Positive correlations (p <0.5) between the percentage reduction of ST of the medium and surfactin concentration (r = 0.44), indicate that this parameter is determinant of the ability to synthesize this compound. The blood agar test has been shown to be useful only as a pre-selection criterion for surfactin producers (18 strains selected by this method reduced ST by only ≤30%). The proposed selection strategy proved effective and made it possible to select the BS15 strain that reduced the ST of the medium to 30.56 ± 0.15 mN/m and simultaneously provided a high concentration of surfactin compared to other strains.

Effects of chemical elements in the trophic levels of natural salt marshes.

The relationships between the bioaccumulation of Na, K, Ca, Mg, Fe, Zn, Cu, Mn, Co, Cd, and Pb, acidity (pH), salinity (Ec), and organic matter content within trophic levels (water-soil-plants-invertebrates) were studied in saline environments in Poland. Environments included sodium manufactures, wastes utilization areas, dumping grounds, and agriculture cultivation, where disturbed Ca, Mg, and Fe exist and the impact of Cd and Pb is high. We found Zn, Cu, Mn, Co, and Cd accumulation in the leaves of plants and in invertebrates. Our aim was to determine the selectivity exhibited by soil for nutrients and heavy metals and to estimate whether it is important in elucidating how these metals are available for plant/animal uptake in addition to their mobility and stability within soils. We examined four ecological plant groups: trees, shrubs, minor green plants, and water macrophytes. Among invertebrates, we sampled breastplates Malacostraca, small arachnids Arachnida, diplopods Diplopoda, small insects Insecta, and snails Gastropoda. A higher level of chemical elements was found in saline polluted areas (sodium manufactures and anthropogenic sites). Soil acidity and salinity determined the bioaccumulation of free radicals in the trophic levels measured. A pH decrease caused Zn and Cd to increase in sodium manufactures and an increase in Ca, Zn, Cu, Cd, and Pb in the anthropogenic sites. pH increase also caused Na, Mg, and Fe to increase in sodium manufactures and an increase in Na, Fe, Mn, and Co in the anthropogenic sites. There was a significant correlation between these chemical elements and Ec in soils. We found significant relationships between pH and Ec, which were positive in saline areas of sodium manufactures and negative in the anthropogenic and control sites. These dependencies testify that the measurement of the selectivity of cations and their fluctuation in soils provide essential information on the affinity and binding strength in these environments. The chemical elements accumulated in soils and plants; however, further flow is selective and variable. The selectivity exhibited by soil systems for nutrients and heavy metals is important in elucidating how these metals become available for plant/animal uptake and also their mobility and stability in soils.

Sex ratio of White Stork Ciconia ciconia in different environments of Poland.

The aim of this study was to analyze the variation in sex ratio of White Stork Ciconia ciconia chicks from differentiated Poland environments. We took under a consideration the impact of Cd and Pb for establish differences among sex ratio in chicks. We also study multiplex PCR employment for establish gender considerations. We collected blood samples via venipuncture of brachial vein of chicks during 2006-2008 breeding seasons at the Odra meadows (SW-Poland; control), which were compared with those from suburbs (SW-Poland), and from copper smelter (S-Poland; polluted) and from swamps near Baltic Sea. We found differences among sex ratio in White Stork chicks from types of environment. Male participation in sex structure is importantly higher in each type of environment excluded suburban areas. Differences in White Stork sex ratio according to the degree of environmental degradation expressed by Cd and Pb and sex-environment-metal interactions testify about the impact of these metals upon sex ratios in storks. Simultaneously, as a result of multiplex PCR, 18S ribosome gene, which served as internal control of PCR, was amplified in male and female storks. It means that it is possible to use primers designed for chicken in order to replicate this fragment of genome in White Stork. Moreover, the use of Oriental White Stork Ciconia boyciana W- chromosome specific primers makes it possible to determine the sex of C. ciconia chicks. Many factors make sex ratio of White Stork changes in subsequent breeding seasons, which depend significantly on specific environmental parameters that shape individual detailed defense mechanisms.

Variable contribution of functional prey groups in diets reveals inter- and intraspecific differences in faecal concentrations of essential and non-essential elements in three sympatric avian aerial insectivores: a re-assessment of usefulness of bird faeces in metal biomonitoring.

Aerial insectivores through their insect diet can contribute to biotransfer of elements across habitats. We investigate the relationship between dietary composition as expressed by the contributions of six functional invertebrate prey groups (primarily of agriculturally subsidised invertebrates characteristic of agricultural areas in temperate regions of Europe) and concentrations of essential (Na, K, Ca, Mg, Fe, Cu, Zn, Mn, Co) and non-essential (As, Cd, Pb) elements of environmental concern in the faeces of nestlings of three species of avian aerial insectivores - Common Swift Apus apus, Barn Swallow Hirundo rustica and House Martin Delichon urbicum - which breed sympatrically and use apparently similar resources of flying insect prey. There were significant differences between the species for 7 of the 12 elements (Ca, Zn, Cu, Co, As, Pb, Cd); these differences were attributable to the variable dietary composition, even though the concentrations of the elements varied enormously between the faecal samples from the individual species. Partial correlation analysis between the biomass (expressed in mg dry weight) of the six functional prey groups and faecal concentrations of elements showed the highest number of significant relationships for toxic metals (As, Pb and Cd). The results of the General Regression Model explaining faecal element concentrations revealed the different explanatory power of the effects of PCA (of six functional prey groups) dietary scores. A significant fit of GRM was obtained for 7 elements (Na, Mg, Fe, Mn, As, Pb, Cd) for Barn Swallows, 2 elements (Cu, As) for House Martins and 1 element (Mn) for Common Swifts. Overall, the results confirmed our predictions that the biomass of consumed coprophilous taxa and insects from crop habitats was positively correlated with the faecal concentrations of toxic elements. Unexpectedly, however, the faecal samples (primarily those of Common Swifts) that contained many oil-seed rape insect pests had lower Ca, Pb and Cd levels and a higher As level. Our study implies that the cross-boundary transfer of contaminants, primarily non-essential elements, by aerially foraging birds through the considerable accumulation of their faeces has potential consequences for the local biogeochemical cycle and environmental quality.

Essential and nonessential elements in nestling rooks Corvus frugilegus from eastern Poland with a special emphasis on their high cadmium contamination.

Concentration of minerals (sodium, potassium, calcium [Ca], magnesium, iron [Fe], copper, zinc [Zn], manganese [Mn], and cobalt) as well as toxic metals (cadmium [Cd], lead [Pb]) were determined in five tissues (liver, lung, kidney, muscle, and bone) of nestling rooks (Corvus frugilegus; 1 to 13 days old) found dead in seven breeding colonies in eastern Poland. Cd concentration in all analyzed tissues was in the narrow range of 17.0-17.2 mg/kg dry weight (dw) Cd, which in the light of the literature data indicates acute contamination by this toxic metal. Similarly, we found increased levels of Pb, which in all tissues ranged between 5.0 and 6.2 mg/kg dw. Results of multivariate general linear model (GLM) testing of the effect of three variables (tissue type, colony, and nestling age) on tissue concentrations of various metals showed significance for Fe, Cu, Zn, and Mn. Only concentrations of Ca, Fe, and Zn differed significantly between the analyzed tissues. GLM analysis did not show any statistically significant differences in tissue levels of minerals and both toxic metals among examined rookeries, which indicates the widespread presence of nonpoint Cd and Pb pollution linked to agricultural activity and similar levels of these inorganic contaminants on crop fields (feeding grounds) around breeding colonies. We concluded that high levels of both toxic metals, Cd and Pb, probably resulting from the diet of nestling rooks, are based mainly on a diet of ground-dwelling beetles gathered on crop fields.

Do toxic heavy metals affect antioxidant defense mechanisms in humans?

The aim of this study was to prove whether anthropogenic pollution affects antioxidant defense mechanisms such as superoxide dismutase (SOD) and catalase (CAT) activity, ferritin (FRT) concentration and total antioxidant status (TAS) in human serum. The study area involves polluted and salted environment (Kujawy region; northern-middle Poland) and Tuchola Forestry (unpolluted control area). We investigated 79 blood samples of volunteers from polluted area and 82 from the control in 2008 and 2009. Lead, cadmium and iron concentrations were measured in whole blood by the ICP-MS method. SOD and CAT activities were measured in serum using SOD and CAT Assay Kits by the standardized colorimetric method. Serum TAS was measured spectrophotometrically by the modified Benzie and Strain (1996) method and FRT concentration-by the immunonefelometric method. Pb and Cd levels and SOD activity were higher in volunteers from polluted area as compared with those from the control (0.0236 mg l(-1) vs. 0.014 mg l(-1); 0.0008 mg l(-1) vs. 0.0005 mg l(-1); 0.137 Um l(-1) vs. 0.055 Um l(-1), respectively). Fe level, CAT activity and TAS were lower in serum of volunteers from polluted area (0.442 g l(-1) vs. 0.476 gl(-1); 3.336 nmol min(-1)ml(-1) vs. 6.017 nmol min(-1)ml(-1); 0.731 Trolox-equivalents vs. 0.936 Trolox-equivalents, respectively), whilst differences in FRT concentration were not significant (66.109 µg l(-1) vs. 37.667 µg l(-1), p=0.3972). Positive correlations between Pb (r=0.206), Cd (r=0.602) and SOD in the inhabitants of polluted area, and between Cd and SOD in the control (r=0.639) were shown. In volunteers from both studied environments TAS-FRT (polluted: r=0.625 vs. control: r=0.837) and Fe-FRT (polluted area: r=0.831 vs. control: r=0.407) correlations, and Pb-FRT (r=0.360) and Pb-TAS (r=0.283) in the control were stated. The higher lead and cadmium concentrations in blood cause an increase of SOD activity. It suggests that this is one of the defense mechanisms of an organism against oxidative stress caused by environmental factors, whilst non-enzymatic mechanisms marked by TAS are the main antioxidant defense system in relation with Pb concentration in humans from unpolluted area. Simultaneously, the higher CAT activity and TAS can indicate that these mechanisms play a key role in the antioxidant protection in non-stressed environments.