Wolfgang Hadnagy

Pyrethroids used indoors--biological monitoring of exposure to pyrethroids following an indoor pest control operation.

A prospective epidemiological study with respect to pyrethroid exposure was carried out combining clinical examination, indoor monitoring and biological monitoring. The results of the biological monitoring are presented. Biological monitoring was performed in 57 persons before (T1) as well as 1 day (T2), 3 days (T3), 4-6 months (T4), and 10-12 months (T5) following a pest control operation (PCO) with pyrethroid containing products such as cyfluthrin, cypermethrin, deltamethrin or permethrin. Pyrethroids in blood were measured by GC-ECD. The respective metabolities cis- and trans-3-(2,2-dichlorovinyl)-2,2-dimethylcyclopropane carboxylic acid (DCCA), cis-3-(2,2-dibromovinyl)-2,2-dimethylcyclopropane carboxylic acid (DBCA), 3-phenoxybenzoic acid (3-PBA) and fluorophenoxybenzoic acid (FPBA) were measured in urine using GC/MS. For all cases the concentrations of pyrethroids in blood were found to be below the detection limit of 5 micrograms/l before and after the PCO. With a detection limit of 0.2 microgram/l of the investigated metabolites, the percentage of positive samples were 7% for cis-DCCA, 3.5% for trans-DCCA and 5.3% for 3-PBA before PCO. One day after PCO (T2) the percentage of positive samples increased remarkably for cis-DCCA (21.5%), trans-DCCA (32.1%) and 3-PBA (25%) showing significantly increased internal doses as compared to pre-existing values. This holds also true for T3, whereas at T4 and T5 the significant increase was no more present. FPBA and DBCA concentrations were below the respective detection limit before PCO and also in most cases after PCO. In 72% of the subjects the route of pyrethroid uptake (measured by determining the DCCA isomeric ratio) was oral/inhalative and in 28% it was dermal. Based on the biological monitoring data it could be shown that appropriately performed pest control operations lead to a significant increase of pyrethroid metabolite concentration in the early phase (1 and 3 days) after pyrethroid application as compared to the pre-exposure values. However, evaluated metabolite concentrations 4-6 months after PCO did not exceed values of published background levels.

Immunological parameters in humans exposed to pesticides in the agricultural environment

Immune parameters were examined in 224 sera of non-exposed controls and in 304 sera of pesticide applicators in the agricultural environment. In comparison to the control group pesticide applicators showed significant increased odds ratios for neopterin and soluble tumor necrosis factor receptor (sTNF RII) and a decreased odds ratio for immunoglobulin M. Obtained results indicate an enhanced macrophage activation and an impaired humoral defense. These alterations have been found to correlate with exposure duration in the group of pesticide applicators in agriculture. For subjects who worked in indoor pest control an inverse correlation for sTNF RII with exposure duration was obtained indicating impairment of cell mediated immune function. It can be concluded that exposure to pesticides in the agricultural environment may contribute to modulation of the immune system. Since immune modulating agents can potentially lead to adverse health consequences the involvement of immune biomarkers in pesticide-related health studies seems to be of considerable value for risk assessment studies.

Induction of mitotic cell division disturbances and mitotic arrest by pyrethroids in V79 cell cultures

Five pyrethroids (fenvalerate, deltamethrin, cypermethrin, permethrin, cyfluthrin) differing in their chemical purity were investigated on their cytotoxic effects, especially on their ability to induce mitotic cell division disturbances using Chinese hamster lung cells of line V79. The colony forming ability (CFA) resulted in distinct differences of the cytotoxic effect of the tested pyrethroids, whereby permethrin was found to be most toxic. With the exception of fenvalerate all tested pyrethroids gave rise to inhibition of cell cycle progression as shown by G2/M-arrest of synchronized V79 cells by flow cytometry as well as by the increase of the mitotic index as evaluated by light microscopy. The mitotic arresting activity could be attributed to the occurrence of abnormal mitotic figures such as initial and full C-metaphases. The results however indicate, that pyrethroids per se do not contribute to the cytotoxic effects but that other factors such as chemical impurities, source as well as manufacturing process and isomer composition may be responsible for the observed cytotoxic effects.

Pyrethroids used indoors--immune status of humans exposed to pyrethroids following a pest control operation--a one year follow-up study.

A multiparametric analysis of immune components was performed in blood and serum of 61 voluntary persons before and after (1 day, 3 days, 4-6 months, 10-12 months) a professional pest control operation (PCO) using pyrethroids. Following parameters were included in the study (1) immunological parameters of the humoral defence, i.e. immunoglobulins of the classes A, G, M and E, complement components C3c and C4, acute phase proteins such as acid alpha 1-glycoprotein, haptoglobin, C-reactive protein; (2) mediators and receptors of immunity, i.e. neopterin, soluble interleukin-2 receptor (sIL-2R), soluble interleukin-6 receptor (sIL-6R), soluble tumor necrosis factor receptor (sTNF RII); (3) immunological markers of the cellular defence, i.e. white blood cell counts and lymphocyte (sub)populations such as total lymphocytes (CD2), mature lymphocytes (CD3), T-helper/inducer cells (CD4), T-suppressor/cytotoxic cells (CD8), B-cells (CD20), natural killer cells (CD56), as well as the ratio of CD4/CD8. The medians of all investigated immune components found before and for all time intervals after pyrethroid application were within the reference interval with respect to the total collective. Within this physiological range the investigated parameters showed a trend to lower values predominantly during the early phase (1 and 3 days) after PCO, partially being significant. Significant decreases were no more present in the late phase (6 to 12 month) after PCO indicating reversibility. Atopics did not differ in the immune response after PCO as compared to non-atopics. Obtained results suggest a modulation of immune components after a correct performed PCO within the physiological range towards lower values during the first days. However these immune changes are considered to be subtle and underlying compensatory mechanisms of immunoregulation.

Immunological alterations in sera of persons living in areas with different air pollution

The present study aimed to investigate immune parameters in sera of adult persons chronically exposed to different degrees of ambient air pollution. As related to air pollution derived from coal mining industry and coke plants, a significantly increased prevalence of cases with abnormally high serum levels of the immunoglobulins IgA and IgM as well as the complement component C3c was found as compared with a less polluted control area, indicating a higher stimulation of acute reactants in combination with a polyclonal immune response. These findings may be attributed to elevated concentrations of airborne particulates, suggesting that permanent exposure to increased levels of airborne particulates leads to chronic irritation of the airways in association with activation of the immune system, which may give rise to an enhanced risk for chronic airway diseases.

A rapid method for detection of nongenotoxic carcinogens of environmental pollutants using synchronized V79 cells and flow cytometry

Synchronized V79 cells were treated before entering mitosis with known and suspicious mitotic arrestants and analyzed by flow cytometry and by light microscopy. Colcemid, nocodazole, vinblastine, diethylstilbestrol, triethyl lead and cadmium sulfate caused a dose dependent mitotic arrest of up to 80%, in comparison with 6% for the controls. Mixtures of polycyclic aromatic hydrocarbons and heterocyclic compounds induced a mitotic arrest of 50%-60%. Extracts of airborne particulates revealed a mitotic arrest of 10%-40%. In contrast, benzoquinone and hydroquinone led to a G2-block rather than to a mitotic arrest. Results of flow cytometry measurements correlated well with those obtained by light microscopy. Cell synchronization in combination with flow cytometry seems to be of considerable value as a rapid method for testing nongenotoxic agents with mitotic arresting activity.

Inhibition of phagocytosis of human macrophages induced by airborne particulates.

Human monocyte-derived macrophages isolated from peripheral blood were treated with different extracts of airborne particulates collected in the highly industrialized Rhine-Ruhr area. All tested extracts showed a substantial impairment of phagocytosis by inhibition of phagocytic activity as well as phagocytic capacity, while cell viability was rather well maintained. Significant reduction of phagocytosis already appeared at a concentration equivalent to extracted particulates from 3.8 m3 air. Having properties of alveolar macrophages, human monocyte-derived macrophage cultures may offer a reliable in vitro model for assessment of pulmonary toxicity by respirable pollutants.

Hemolytic activity of crystalline silica--separated erythrocytes versus whole blood.

Whole blood and 1% erythrocyte suspensions were treated with crystalline silica (quartz DQ12, Min-U-Sil5) at concentrations of 0.5, 1, 2, and 5 mg/ml. Quartz DQ12 and Min-U-Sil5 revealed a strong dose-dependent hemolytic activity in the 1% erythrocyte suspension reaching nearly total hemolysis (> 80%) at the highest tested concentration of 5 mg/ml. This effect may be ascribed to surface reactivity by silanol groups. In contrast, using whole blood cultures the tested silica dusts caused no or only minor hemolytic activity (< 4%). The mechanism by which the hemolytic activity is prevented in whole blood cultures can be attributed to a number of factors such as the presence of metal binding proteins and free radical scavenger, antioxidant mechanisms and to coating of the silica surface by proteins, antibodies and complement. In contrast to separated erythrocytes whole blood represents an independent physiological compartment with functions of host defence and regulatory functions against cell damaging effects produced by oxidative stress.

Cytokines detectable in saliva of children as appropriate markers of local immunity of the oral cavity: an approach for the use in air pollution studies

The objective of this study was the detection of proinflammatory markers in saliva to be involved in local immunity of the oral cavity. Therefore saliva of 167 schoolchildren aged 8-10 years were investigated for the presence of interleukin-8 (IL-8), tumour necrosis factor alpha (TNF alpha) and soluble tumour necrosis factor receptor type II (sTNFRII). In saliva of schoolchildren sufficient quantities of IL-8 (302.3-4208.6 pg/ml), TNF alpha (0.3-40.6 pg/ml) and sTNFRII (17.6-931.3 pg/ml) were detectable. IL-8, TNF alpha and sTNFRII revealed significant correlations with each other. Results suggest an immunoregulatory mechanism of IL-8, TNF alpha and TNF-receptor to be of special concern in host defence as well as in maintaining homeostasis of local immunity within the oral cavity. Saliva provides an ideal medium for the detection of proinflammatory markers of the oral cavity with respect to mucosal and granulotype origin and may be employed in air pollution epidemiology, especially with regard to children.

Soluble tumor necrosis factor receptor (sTNF RII) in sera of children and traffic-derived particulate air pollution.

Tumor necrosis factor receptor (sTNF RII) was determined in sera of 160 healthy schoolchildren of the city of Düsseldorf, Germany, living in areas with different traffic density. According to the frequency distribution a higher prevalence of children with increased sTNF RII values (> 3000 pg/ml) were found for a high traffic area as compared to a low traffic area. Based on sTNF RII values above the 75% percentile of children from the low traffic area, the group of children from the high traffic area revealed a significant increased odds ratio of 2.5. Concerning traffic-derived particulate air pollution an association between the concentration of fine particles (PM2.5) and sTNF RII serum levels could be observed for both areas. Furthermore, sTNF RII values gave a significant positive correlation with C3c, an activation product of the complement component C3. C3c has been shown to be a sensitive indicator of the non-specific humoral defence in response to air pollution. Therefore, the results suggest that traffic-derived fine particles may upon inhalation trigger immune modulation via the activation of macrophages and enhanced cytokine production.