Adverse Effects of Non-Metallic Nanoparticles in the Central Nervous System.

The interest in nanoparticles (NPs) and their effects on living organisms has been continuously growing in the last decades. A special interest is focused on the effects of NPs on the central nervous system (CNS), which seems to be the most vulnerable to their adverse effects. Non-metallic NPs seem to be less toxic than metallic ones; thus, the application of non-metallic NPs in medicine and industry is growing very fast. Hence, a closer look at the impact of non-metallic NPs on neural tissue is necessary, especially in the context of the increasing prevalence of neurodegenerative diseases. In this review, we summarize the current knowledge of the in vitro and in vivo neurotoxicity of non-metallic NPs, as well as the mechanisms associated with negative or positive effects of non-metallic NPs on the CNS.

The influence of silver nanoparticles on the process of epithelial transition in the context of cancer metastases.

BACKGROUND: Exposure to nanoparticles (NPs) can occur in a variety of occupational situations. Ultrafine particles of natural and anthropological origin toxicity has been described in epidemiological studies. Meanwhile, the risks associated with NPs exposure are not comprehensively assessed. A wide spectrum of NPs toxicity has been demonstrated, mainly through the induction of oxidative stress and inflammatory mediators. Among the newly described mechanisms of NPs toxicity is the induction of fibrosis via the epithelial-mesenchymal transition (EMT), which is also a key mechanism of cancer metastasis. The effect of NPs on EMT in the context of metastasis has not been sufficiently described so far, and the results of studies do not allow for the formulation of unambiguous conclusions. Therefore, the aim of the work was to determine the biological activity of silver NPs against MDA-MB-436 triple-negative breast cancer cells. MATERIAL AND METHODS: Exposure to nanoparticles (NPs) can occur in a variety of occupational situations. Ultrafine particles of natural and anthropological origin toxicity has been described in epidemiological studies. Meanwhile, the risks associated with NPs exposure are not comprehensively assessed. A wide spectrum of NPs toxicity has been demonstrated, mainly through the induction of oxidative stress and inflammatory mediators. Among the newly described mechanisms of NPs toxicity is the induction of fibrosis via the epithelial-mesenchymal transition (EMT), which is also a key mechanism of cancer metastasis. The effect of NPs on EMT in the context of metastasis has not been sufficiently described so far, and the results of studies do not allow for the formulation of unambiguous conclusions. Therefore, the aim of the work was to determine the biological activity of silver NPs against MDA-MB-436 triple-negative breast cancer cells. RESULTS: Silver nanoparticles (AgNPs) cause a statistically significant increase in relative expression of all tested mesenchymal EMT markers - cadherin 2, vimentin, matrix metalloproteinase 2 and matrix metalloproteinase 9. At the same time, reduction of epithelial cadherin 1 expression was observed. The level of MDA-MB-436 migration and TGF-beta 1 secretion was slighty increased in AgNPs-treated cells, with no influence on invasion potential. CONCLUSIONS: Potentially prometastatic effect of AgNPs encourages further work on the safety of nanomaterials. Med Pr Work Health Saf. 2023;74(6):541-8.

Cold adapted and closely related mucoraceae species colonise dry-aged beef (DAB).

The dry ageing is a historically relevant method of meat preservation, now used as a way to produce the dry-aged beef (DAB) known for its pronounced flavour. Partially responsible for the taste of the DAB may be various microorganisms that grow on the surface of the meat. Historically, the fungal species colonising the DAB were described as members of the genera Thamnidium and Mucor. In this study we used both culture based approach as well as ITS2 rDNA metabarcoding analysis to investigate the fungal community of the DAB, with special emphasis on the mucoralean taxa. Isolated fungi were members of 6 different species from the family Mucoraceae, belonging to the genera Mucor and Helicostylum. Metabarcoding data provided supplementary information regarding the presence of other fungi including those from the Thamnidium genus. In both approaches used in this study isolates closely related to the Mucor flavus strain CBS 992.68 dominated.

Exposure to Chlorpyrifos Alters Proliferation, Differentiation and Fatty Acid Uptake in 3T3-L1 Cells.

Organophosphorus pesticides (OPs) are important factors in the etiology of many diseases, including obesity and type 2 diabetes mellitus. The aim of this study was to investigate the effect of a representative of OPs, chlorpyrifos (CPF), on viability, proliferation, differentiation, and fatty acid uptake in 3T3-L1 cells. The effect of CPF exposure on preadipocyte proliferation was examined by the MTT, NR, and BrdU assays. The impact of CPF exposure on the differentiation of preadipocytes into mature adipocytes was evaluated by Oil Red O staining and RT-qPCR. The effect of CPF on free fatty acid uptake in adipocytes was assessed with the fluorescent dye BODIPY. Our experiments demonstrated that exposure to CPF decreased the viability of 3T3-L1 cells; however, it was increased when the cells were exposed to low concentrations of the pesticide. Exposure to CPF inhibited the proliferation and differentiation of 3T3-L1 preadipocytes. CPF exposure resulted in decreased lipid accumulation, accompanied by down-regulation of the two key transcription factors in adipogenesis: C/EBPα and PPARγ. Exposure to CPF increased basal free fatty acid uptake in fully differentiated adipocytes but decreased this uptake when CPF was added during the differentiation process. Increased free fatty acid accumulation in fully differentiated adipocytes may suggest that CPF leads to adipocyte hypertrophy, one of the mechanisms leading to obesity, particularly in adults. It can therefore be concluded that CPF may disturb the activity of preadipocytes and adipocytes, although the role of this pesticide in the development of obesity requires further research.

Influence of chlorpyrifos exposure on UVB irradiation induced toxicity in human skin cells.

BACKGROUND: Although chlorpyrifos (CPS) has been banned in many developed countries, it still remains one of the best-selling pesticides in the world. Widespread environmental and occupational exposure to CPS pose a serious risk to human health. Another environmental factor that can adversely affect human health is ultraviolet radiation B (UVB, 280-315 nm wave length). Here we attempt determine if exposure to CPS can modify toxic effects of UVB. Such situation might be a common phenomenon in agriculture workers, where exposure to both factors takes place. METHODS: Two skin cell lines; namely human immortalized keratinocytes HaCaT and BJ human fibroblasts were used in this study. Cytotoxicity was investigated using a cell membrane damage detection assay (LDH Cytotoxicity Assay), a DNA damage detection assay (Comet Assay), an apoptosis induction detection assay (Apo-ONE Homogeneous Caspase-3/7 Assay) and a cell reactive oxygen species detection assay (ROS-Glo H2O2 assay). Cytokine IL-6 production was also measured in cells using an ELISA IL-6 Assay. RESULTS: Pre-incubation of skin cells with CPS significantly increased UVB-induced toxicity at the highest UVB doses (15 and 20 mJ/cm2). Also pre-exposure of BJ cells to CPS significantly increased the level of DNA damage, except for 20 mJ/cm2 UVB. In contrast, pre-exposure of HaCaT cells, to CPS prior to UVB radiation did not cause any significant changes. A decrease in caspase 3/7 activity was observed in HaCaT cells pre-exposed to 250 µM CPS and 5 mJ/cm2 UVB. Meanwhile, no statistically significant changes were observed in fibroblasts. In HaCaT cells, pre-exposure to CPS resulted in a statistically significant increase in ROS production. Also, in BJ cells, similar results were obtained except for 20 mJ/cm2. Interestingly, CPS seems to inhibited IL-6 production in HaCaT and BJ cells exposed to UVB (in the case of HaCaT cells for all UVB doses, while for BJ cells only at 15 and 20 mJ/cm2). CONCLUSIONS: In conclusion, the present study indicates that CPS may contribute to the increased UVB-induced toxicity in skin cells, which was likely due to the induction of ROS formation along with the generation of DNA damage. However, further studies are required to gain better understanding of the mechanisms involved.

Effect of silver nanoparticles on cytotoxicity, oxidative stress and pro-inflammatory proteins profile in lung adenocarcinoma A549 cells.

INTRODUCTION AND OBJECTIVE: The general population is exposed to silver nanoparticles (AgNPs) released into the environment, e.g. through the respiratory tract. Lung cancers are among the most frequently diagnosed and deadly malignancies, often diagnosed at late stage with existing distant metastases. The aim of the study was to determine the activity of AgNPs against A549 lung cancer cells. MATERIAL AND METHODS: A549 cells and AgNPs were used in the study. Cytotoxicity was tested by MTT and NR assays. Oxidative stress was determined by measuring malonyldialdehyde and level of free -SH groups Proteins secretion was assessed using the Human Profiler Cytokine Array Kit assay. RESULTS: AgNPs reduce A549 cells viability and induce oxidative stress. They also lead to increased secretion of several proinflammatory proteins, which stimulate metastasis. CONCLUSIONS: AgNPs exhibit direct anti-cancer effect, however, their potentially promethastic effect encourages further work on the safety of nanomaterials.

Fungal Biostarter Effect on the Quality of Dry-Aged Beef.

Meat aging is a process consisting of its storage in specific conditions which leads to an increase in its organoleptic qualities. The most common method of meat aging is the wet vacuum-bag based method, whereas the traditional method, called dry-aging, involves keeping meat at a low temperature for an extended time. However, this process is characterized by low repeatability of the results. Therefore, different approaches to stabilize the process are tested. The aim of this study was to analyze the influence of the Mucor flavus biostarter on the physicochemical characteristics and sensory quality of dry-aged beef (DAB). We hypothesized that a fungal biostarter positively influences the quality of DAB and stabilizes the dry-aging process. Meat control samples (N = 7) and samples inoculated with the Mucor flavus biostarter (N = 7), originating from 14 individuals crossbred from Holstein-Friesian cows with bulls of meat breeds, were put into the dry-aging fridge (DryAger, Bad Saulgau, Germany) for 28 days. The physicochemical parameters (pH, color parameters, WHC, GP (glycolytic potential), chemical composition of muscle, the content of malondialdehyde, shear force), muscular protein proteolysis (SDS-PAGE), sensory quality, and microbial composition of DAB were assessed after aging. The results showed a significant effect of the fungal biostarter on pH increase (0.25 units), and light myosin chain proteolysis (approximately 16%) as well as improvement of sensory quality (e.g., acceptability was improved by one unit in an applied scale 1-9). All together, the M. flavus-based biostarter was shown to increase the quality of DAB.

ThermoSteg-Covert Channel for Microbolometer Thermographic Cameras.

The article presents a new concept-steganography in thermography. Steganography is a technique of hiding information in a non-obvious way and belongs to sciences related to information security. The proposed method, called ThermoSteg, uses a modification of one of the parameters of the thermal imaging camera-integration time-to embed the signal containing hidden information. Integration time changing makes the microbolometer array heat up while reading the sensors. The covert information can be extracted from the stream of thermograms recorded by another thermal camera that observes the first one. The covert channel created with the ThermoSteg method allows the transmission of covert data using a thermal sensor as a wireless data transmitter. This article describes a physical phenomenon that is exploited by the ThermoSteg method and two proposed methods of covert data extraction, and presents the results of experiments.

StegoFrameOrder-MAC Layer Covert Network Channel for Wireless IEEE 802.11 Networks.

The proposed StegoFrameOrder (SFO) method enables the transmission of covert data in wireless computer networks exploiting non-deterministic algorithms of medium access (such as the distributed coordination function), especially in IEEE 802.11 networks. Such a covert channel enables the possibility of leaking crucial information outside secured network in a manner that is difficult to detect. The SFO method embeds hidden bits of information in the relative order of frames transmitted by wireless terminals operating on the same radio channel. The paper presents an idea of this covert channel, its implementation, and possible variants. The paper also discusses implementing the SFO method in a real environment and the experiments performed in the real-world scenario.

Chlorpyrifos alters expression of enzymes involved in vitamin D3 synthesis in skin cells.

Skin acts as a mechanical barrier between human body and environment. Epidermal cells are regularly exposed to many physiological and environmental stressors, such as pesticides, like chlorpyrifos (CPS). It is recognised that CPS may affect metabolism of other exo- and endogenous substances by affecting enzyme activity and expression. This study aims to investigate the effect of CPS on expression of CYP27A1, CYP27B1 and CYP24A1, the enzymes involved in synthesis and metabolism of vitamin D3, in human keratinocytes HaCaT and human fibroblasts BJ. Synthesis of vitamin D3 in cells was initiated by irradiating with UVB. Expression of CYP27A1, CYP27B1 and CYP24A1 was evaluated by RT-qPCR and Western blot. Our experiments revealed that expression of all tested cytochrome P450 isoforms in cells exposed to CPS changed significantly. Exposure of HaCaT keratinocytes to CPS decreased CYP27A1 mRNA levels, but increased CYP27B1 and CYP24A1 mRNA levels. This was confirmed at the protein level, except for the CYP27A1 expression. Outcome for the BJ cells was however less conclusive. Though exposure to CPS decreased CYP27A1 and CYP27B1 mRNA levels, at protein level increasing concentration of CPS and UVB intensity induced expression of CYP27A1 and CYP24A1. The expression of CYP27B1 isoform decreased in line with mRNA level. Nevertheless, it can be concluded that CPS may therefore interrupt vitamin D3 metabolism in skin cells, but further studies are required to better understand such mechanisms.

Nuclear Factor kappa B activation by Ag, Au nanoparticles, CdTe quantum dots or their binary mixtures in HepG2 cells.

INTRODUCTION AND OBJECTIVE: Nuclear factor kappa B (NF-κB) signalling pathway plays a central role in the regulation of cellular response to stress. The aim of the study was to investigate the ability of silver nanoparticles (AgNPs), gold nanoparticles (AuNPs), CdTe quantum dots (CdTeQDs) or their binary mixtures to stimulate NF-κB binding in HepG2 cells. A dual luciferase reporter system was used to investigate NF-κB binding. MATERIAL AND METHODS: Cells were transiently transfected with a firefly luciferase reporter system and Renilla luciferase expression plasmid as a transfection efficiency control. Twenty- four hours after transfection, the cells were treated with nanoparticles (10 µg/cm3 AgNPs, 10 µg/cm3 AuNPs, 3 µg/cm3 CdTeQDs) or with 10 ng/cm3 TNFα as a positive control. Six hours later, the cells were lysed and the activities of the luminescence of firefly and Renilla luciferases were measured using the Dual-Luciferase Reporter Assay System. RESULTS: AuNPs and CdTeQDs alone significantly inhibited NF-κB binding activity. Co-treatment with AgNPs and CdTeQDs resulted in an additive effect, whereas the presence of AgNPs diminished the inhibitory effect of AuNPs. Interestingly, significant antagonism was observed between AuNPs and CdTeQDs, suggesting a similar mode of action. CONCLUSIONS: Comparison of the NF-κB binding activity induced by the mixtures of NPs suggests that in some cases NF-κB binding activity might differ from that observed for the NPs alone.

Two Sides to the Same Coin-Cytotoxicity vs. Potential Metastatic Activity of AgNPs Relative to Triple-Negative Human Breast Cancer MDA-MB-436 Cells.

Silver nanoparticles (AgNPs) are used in many fields of industry and medicine. Despite the well-established antimicrobial activity, AgNPs are foreseen to be used as anticancer drugs due to the unusual feature-inability to induce drug resistance in cancer cells. The aim of the study was to assess biological activity of AgNPs against MDA-MB-436 cells. The cells were derived from triple-negative breast cancer, a type of breast cancer with poor prognosis and is particularly difficult to cure. AgNPs were toxic to MDA-MB-436 cells and the probable mechanism of toxicity was the induction of oxidative stress. These promising effects, giving the opportunity to use AgNPs as an anti-cancer agent should, however, be treated with caution in the light of further results. Namely, the treatment of MDA-MB-436 cells with AgNPs was associated with the increased secretion of several cytokines and chemokines, which were important in breast cancer metastasis. Finally, changes in the actin cytoskeleton of MDA-MB-436 cells under the influence of AgNPs treatment were also observed.

Organophosphorus pesticides can influence the development of obesity and type 2 diabetes with concomitant metabolic changes.

Widespread use and the bioaccumulation of pesticides in the environment lead to the contamination of air, water, soil and agricultural resources. A huge body of evidence points to the association between the pesticide exposure and increase in the incidence of chronic diseases, e.g. cancer, birth defects, reproductive disorders, neurodegenerative, cardiovascular and respiratory diseases, developmental disorders, metabolic disorders, chronic renal disorders or autoimmune diseases. Organophosphorus compounds are among the most widely used pesticides. A growing body of evidence is suggesting the potential interdependence between the organophosphorus pesticides (OPs) exposure and risk of obesity and type 2 diabetes mellitus (T2DM). This article reviews the current literature to highlight the latest in vitro and in vivo evidences on the possible influence of OPs on obesity and T2DM development, as well as epidemiological evidence for the metabolic toxicity of OPs in humans. The article also draws attention to the influence of maternal OPs exposure on offspring. Summarized studies suggest that OPs exposure is associated with metabolic changes linked with obesity and T2DM indicated that such exposures may increase risk or vulnerability to other contributory components.

Assessment of DNA damage in Polish children environmentally exposed to pesticides.

Exposure to pesticides leads to complex, long-lasting adverse effects on human health, and poses a substantial risk to those living in areas devoted to agriculture. Children are particularly vulnerable to the pesticide exposure, due to the developmental, dietary and physiological factors. Small body mass and typical exploratory behavior result in increased risk of intoxication. Thus, even exposure to low concentrations of pesticides, if of sufficient duration, may lead to permanent health disorders and limit their harmonious development. In this study 108 children, living in areas of an intense pesticide use and a control group (n = 92) of children from an agrotouristic area were investigated, whether DNA damage increased due to prolonged pesticide exposure. A presence of DNA breaks and oxidative damage to DNA bases, characterized as Fpg-sensitive sites, were detected by comet assay. Micronuclei (MN) formation was evaluated by cytokinesis-block MN assay. The exposure of children to pesticides resulted in increased number of MN in peripheral blood lymphocytes (P = 0.016), increased DNA strand breaks level (P = 0.002) and oxidative damage to DNA (P < 0.001). Negative correlation was demonstrated between the level of DNA strand breaks and acetylcholinesterase (AChE) activity in exposed group. In conclusion, despite just environmental pesticide exposure in the test group of children, significant biological effects were detected.

IL­6 prevents CXCL8­induced stimulation of EpCAM expression in ovarian cancer cells.

Epithelial cell adhesion molecule (EpCAM), which is expressed in the majority of epithelial tissues, exhibits tumor growth promoting abilities and is overexpressed in human epithelial ovarian cancer. Therefore, EpCAM is considered to be a promising target for specific immune­based therapies. The present study evaluated the role of IL­6 and IL­8 in the expression of EpCAM in the A2780 human ovarian cancer cell line. Furthermore, the cellular localization of the EpCAM protein in A2780 cells was determined and the effect of EpCAM inhibition on the proliferation of the A2780 cells was investigated. An MTT assay demonstrated that blocking EpCAM with anti­EPCAM antibodies had no effect on cellular metabolic activity (proliferation). Gene expression analysis revealed that IL­8 increased EpCAM expression, whereas IL­6 and the combination of IL­6/IL­8 had no effect on EpCAM expression. Immunofluorescence analysis confirmed that EpCAM is expressed on A2780 cell membranes. The present results demonstrated that IL­8 increased EpCAM expression at the mRNA level in ovarian cancer cells and suggested a potential role of IL­6 as an inhibitor of IL­8­stimulated EpCAM expression.

Chlorpyrifos stimulates expression of vitamin D3 receptor in skin cells irradiated with UVB.

Skin, the organ responsible for vitamin D synthesis, is fully exposed to many xenobiotics, e.g. polycyclic aromatic hydrocarbons and pesticides. A broad spectrum organophosphorus insecticides (OP's), such as chlorpyrifos (CPS), are commonly used in agriculture and to control domestic insects. Thus, the aim of this study was to investigate the effect of chlorpyrifos, on the expression of vitamin D3 receptor (VDR) in human keratinocytes cell line HaCaT and fibroblasts cell line BJ. The impact of CPS and UVB radiation on cell viability were examined by Neutral Red assay. The effect of CPS on VDR expression was evaluated by RT-qPCR and flow cytometry (FC). The presented study demonstrated that exposure to CPS and UVB significantly affects the viability of HaCaT and BJ cells lines. Results also revealed that exposure to CPS induced the expression at mRNA and protein level of VDR nuclear receptor in both cell lines exposed to UVB. In HaCaT incubated with 250 µM CPS and 15 mJ/cm2 UVB, the relative VDR expression was ∼2-fold higher; whereas in BJ incubated with 250 µM CPS and 20 mJ/cm2, UVB was∼3-fold higher. Results from FC confirmed this result, as VDR expression increased by ~250% in HaCaT incubated with 250 µM CPS and 20 mJ/cm2 UVB, and in BJ incubated with 250 µM CPS, and 20 mJ/cm2 UVB cells VDR expression increased by ~190%, compared with control. It can therefore be concluded that OPs pesticide might interfere with vitamin D3 metabolism in skin cells.

Event-related potentials (ERP) and SGIP1 gene polymorphisms in alcoholics: relation to family history of alcoholism and drug usage.

OBJECTIVE: The electrophysiological characteristics may serve as valuable biomarkers for the genetic vulnerability underlying alcoholism. The purpose of this study was to evaluate the potential associations between single nucleotide polymorphisms (SNPs) located in the SGIP1 gene and the theta ERP quantitative traits. METHOD: The theta band (4-7 Hz) visual ERP occurring in the P300 response in the resting EEG were examined to explore the electrophysiological effects of alcohol on the brain in five regions: frontal, central, parietal, temporal and occipital in patients with alcohol addiction. In addition, we tested the potential associations between single nucleotide polymorphisms (SNPs) located in the SGIP1 gene and ERP quantitative traits. RESULTS: We found that the amplitude of the auditory P300 response differed considerably among groups of alcoholics in the frontal, central and temporal areas of the brain and it was lower in the studied brain regions in alcoholics in comparison to non-alcoholics. However, among subjects in the young adult group (GR-1) there was no statistical difference in amplitude of P300 response with control subjects in all studied brain regions in comparison with non-alcoholics. Moreover, we revealed that SNP rs10889635 had a significant effect on P300 amplitude in the central and temporal regions. The reduced P300 amplitude was in AA carriers in comparison to both carriers of GG and GA alleles. CONCLUSION: The present study demonstrated a possible association of target P300 evoked theta and of alcohol dependence with SNPs from the gene SGIP1 in the region of rs10889635, but further studies are required.

Effect of IL-6 and IL-8 on the expression of the complement activation inhibitors MAC-inhibitory protein and decay-accelerating factor in ovarian cancer A2780 cells.

The aim of the present study was to evaluate the role of interleukin (IL)-6 and IL-8 on the expression of the membrane-bound complement inhibitors membrane attack complex-inhibitory protein (CD59) and decay-accelerating factor (CD55), in the human ovarian carcinoma A2780 cell line, which is a non-producing IL-6 cell line that does exhibit IL-6 responsiveness, due to the presence of IL-6 receptors. Extracellular levels of complement system inhibitors were evaluated by western blotting and reverse transcription-quantitative polymerase chain reaction. Cellular localization of CD55 and CD59 in the ovarian cancer cells was assessed by immunofluorescence. The detection of a soluble form of CD55 and CD59 released by the A2780 cells following stimulation with IL-6 and IL-8 was detected by enzyme-linked immunosorbent assay. The present data revealed that A2780 cells express CD55 and CD59 at the mRNA and protein level, but do not secrete these proteins to the culture medium. Results of western blotting demonstrated that the protein level of CD59 was regulated by IL-6 and IL-8 in a dose-dependent manner. Immunofluorescence analysis revealed that the ovarian cancer A2780 cell line expresses the membrane bound form of CD55 protein. The present results indicate that CD55 and CD59 may affect the efficiency of complement-mediated immunotherapies.

IL­6 and IL­8 enhance factor H binding to the cell membranes.

The aim of the present study was to assess the role of interleukin (IL)­6 and IL­8 on the expression of fluid­phase complement inhibitor, factor H (FH), and FH­like protein 1 (FHL­1), in the A2780 ovarian carcinoma cell line. This cell line does not normally produce IL­6, however, is IL­6 responsive due to the presence of receptor for IL­6. The presence of FH and FHL­1 in the cell lysates was confirmed by western blotting. The levels of FH and FHL­1 in the medium were determined by enzyme­linked immunosorbent assay. To evaluate gene expression, reverse transcription­quantitative polymerase chain reaction was performed. The cellular localization of FH and FHL­1 in ovarian cancer cells was assessed by immunofluorescence. The present study revealed that FH, contrary to FHL­1, was secreted by ovarian cancer cells, however, this process was independent of IL stimulation. No significant differences were observed in the concentration of FH in the control cells, when compared with the samples treated with IL­6/IL­8. The results of western blotting revealed that the protein expression levels of FH and FHL­1 were not regulated by IL­6 and IL­8 in a dose­dependent manner. Immunofluorescence analysis confirmed that the A2780 ovarian cancer cell line expressed both membrane bound and intracellular forms of FH and FHL­1. The present data revealed that the A2780 cells expressed and secreted FH protein and are also able to bind FH and FHL­1. This may influence the efficiency of complement mediated immunotherapy.

Cholinesterase activity in blood and pesticide presence in sweat as biomarkers of children`s environmental exposure to crop protection chemicals.

INTRODUCTION: On the contrary to the adult population exposed to pesticides, mostly on occupational basis, rural children are mostly exposed to pesticides deposited in the environment. However, even this constant, distributed in time exposure to low concentrations of pesticides may led to permanent health disorders and limit children's harmonious development. OBJECTIVE: The main objective of the study was to evaluate the usefulness of aacetylcholinesterase (AChE) and butyrylcholinesterase (BuChE) activity determination as a marker of children's environmental exposure to pesticides. An additional aim was to evaluate the usefulness of sweat patches as a novel, non-invasive method of detection of pesticides in sweat as a measure of pesticide exposure. MATERIALS AND METHOD: A total of 108 children living in areas of intense pesticide use, and as a control group, 92 children living in an agro-tourist area were enrolled in the study. The AChE and BuChE activity was assayed colorimetricaly in diluted whole blood or plasma, respectively. In addition, selected pesticides were measured by GC/MS analysis in samples of the subject's sweat absorbed onto a sorbent. RESULTS: The study demonstrated significantly lower AChE and BuChE activity, respectively, in the diluted whole blood and plasma of children exposed to pesticides, compared to the control group (p<0.001 and p=0.003, respectively). The measured mean level of AChE activity was 241.63 ± 26.76 and 348.0 ± 46.95 mU/µmolHb in the exposed and the control group, respectively, whereas the mean activity of BuChE was 424.1 ± 81.1 and 458.6 ± 86.5 mmol/L/min. In addition, pesticide metabolites were detected in 19 (17.6%) sweat samples collected from exposed children. CONCLUSIONS: Altogether, the study indicated that cholinesterase activity is a sensitive marker of the children's environmental exposure to pesticides, whereas sweat patches are useful devices for collecting samples to be analysed for the presence of the pesticides.