Integrated Exposure Assessment Survey (INES) exposure to persistent and bioaccumulative chemicals in Bavaria, Germany.

The Integrated Exposure Assessment Survey (INES) was started in the year 2005. Altogether 50 healthy adults living in Bavaria, Germany, were included into the study. Monitoring was conducted in accordance with relevant routes of human exposure (inhalation, ingestion) and integrated different pathways (indoor air, food, house dust). This approach consisted of a combination of external measurements of contaminants with the determination of these substances or their metabolites in body fluids. The target substances were phthalates, perfluorinated compounds (PFC), polychlorinated biphenyls (PCBs), polychlorinated dibenzo-p-dioxins (PCDDs) and polychlorinated dibenzofurans (PCDFs), and polybrominated diphenylethers (PBDEs). This paper gives a brief description of the objectives and the concept of INES as well as methods of sampling and analyses of target compounds. Some preliminary results of biomonitoring data for PFC and phthalates as well as of the dietary intake of DEHP will be discussed.

[Differences in small area prevalence of respiratory diseases in children: challenges of an investigation initiated by parents]. / Kleinräumige Prävalenzunterschiede von Atemwegserkrankungen bei Kindern: Grenzen der Erkenntnismöglichkeiten einer von Eltern angeregten Studie.

OBJECTIVE: The aim of our study was to assess the prevalence of respiratory diseases in children living in a rural area in Bavaria in a systematic and standardised way to clarify whether a disease cluster exists. Reports from a parents' initiative claiming an increase of diseased children and assuming an association with air pollution due to nearby industrial sources and heavy traffic were the reason for the study. METHODS: Parents of all children living in small towns specified by the parents' initiative as affected region (central area) and parents of those children aged 6-7 or 13-14 years living in adjacent areas (control area) were asked to complete a written questionnaire. In bivariate analysis, differences in prevalences between the central area and the control area were assessed by the chi (2) test. In multivariate analysis, confounder-adjusted odds ratios with 95 % confidence intervals were calculated by logistic regression. In addition, generalized linear mixed models were used to control for potential clusters in families. RESULTS: The response rate was rather low (total 46 %, central area 53 %, control area 34 %). The study population comprised 121 girls and 141 boys. There was a pattern of increased prevalences of cough, wheeze and respiratory diseases caused by infections in children living in the central area compared to the control area. However, statistical significance was rarely observed. No systematic differences were observed for the prevalence of allergic rhinitis and atopic dermatitis. Selection bias due to varying response rates in the central versus the control area and information bias introduced by the preceding intensive public discussion could not be excluded. The comparison of prevalences detected in the small area with published data from other epidemiological studies in Germany did not indicate a systematically increased prevalence of respiratory symptoms or physician-diagnosed respiratory diseases in children from the whole area as well as in children from the central area only. CONCLUSION: This study exemplifies the challenges of small area investigations initiated by the residential community to clarify an assumed disease cluster caused by environmental pollution. The prior comparison of small area prevalence data with results of other epidemiological studies aids the decision making as to whether a detailed study with comprehensive assessment of individual exposure is justifiable.

[Older soil contaminations and human biomonitoring--two typical examples]. / Altere Bodenbelastungen und Human-Biomonitoring--zwei Fallbeispiele.

AIM: A health risk assessment of inhabitants had to be accomplished in two areas of former color production works, which had caused considerable soil contaminations predominantly with arsenic and lead. METHODS: Samples of soil, blood and urine were analysed during orientation and detail investigations according to the German soil protection laws. RESULTS: The soil investigations revealed high contaminations in some parts of the two areas (maximum concentrations 550 mg arsenic/kg and 9100 mg lead/kg in residential topsoil from depths of 0-35 cm). Contrary to these findings, arsenic levels in the urine and lead levels in the blood of most of the residents were lower than the reference values for the common German population (arsenic 15 microg/L, lead 50-90 microg/L). In some cases of elevated biological concentrations, other sources of exposure could be detected like drinking tea from a tea-pot with a lead-containing glaze or drinking water from a highly contaminated garden well; interventional measures resulted in major decreases in the internal loads of these people. CONCLUSIONS: The investigations clearly showed that an exposure via the pathway soil--water--human can be of serious health concern whereas older soil contaminations are seemingly of low relevance with regard to the direct contact of persons during daily life on those areas. Therefore, it is recommended to increase the soil examination values ("Prüfwerte") of the German soil protection laws by a default factor of 2 or 3 in cases of soil contaminations older than 20 years.

Foreign-body reaction to the bioabsorbable suretac device.

SUMMARY: Arthroscopic shoulder stabilization with the bioabsorbable Suretac device (Acufex Microsurgical, Mansfield, MA) offers some technical advantages compared with other approaches. However, in 4 patients (3 men, 1 woman; between 20 and 35 years of age) with 3 SLAP lesions grades 2 and 1 post-traumatic anterior shoulder instability, breakage and early loosening of the Suretac device was observed. All patients complained about shoulder pain and loss of active and passive motion. The arthroscopic examination revealed a massive synovitis without positive cultures. Loose fragments of the Suretac device spread in the joint cavity and induced a foreign-body reaction. Histologically, there was a massive infiltration of phagocytic cells (histiocytes, multinucleated giant cells) and birefringent polymeric particles surrounded by or within histiocytes and multinucleated giant cells. All patients underwent arthroscopic synovectomy. Because of instability, 2 of them required restabilization, which was performed with suture anchors (Fastak [Arthrex, Naples, FL], Mitek [Mitek Division, Ethicon, Norderstedt, Germany]). Postoperatively all patients were pain free, progressing to full active and passive range of motion. Our patients suffered from a mechanical failure rather than from a predisposition to exaggerated inflammatory response. According to this observation, the Suretac device seems to be prone to early failure in patients with SLAP lesions because of its biodegradability.

[Anterior cruciate ligament-plasty and high valgus tibial osteotomy as a combined procedure in anterior instability and varus deviation]. / Vordere Kreuzbandplastik und valgisierende hohe Tibiaosteotomie als kombiniertes Vorgehen bei anteriorer Instabilität und Varusmorphotyp.

Besides patients having an acute rupture of the ACL with a clear therapeutical strategy there are increasingly young patients with chronical anterior instability and degenerative arthritis of the medial compartment and varus malalignment. This constellation is taxing severely the operative procedure considering that there is not only the instability which has to be treated but that also an improvement of the arthritis symptomatology has to be achieved. In this paper we present a therapeutical concept of high tibial osteotomy combined with an arthroscopic assisted reconstruction of the ACL which is performed as a single procedure since the early 80-ies. The goal is to remove two severe arthrogenic factors correcting the instability and the pathological anatomical axis, to reduce the medial knee pain and to improve the use of the knee in life activities.

[Arthroscopic shoulder stabilization. Differentiated treatment strategy with Suretac, Fastak, Holmium: YAG-laser and electrosurgery]. / Arthroskopische Schulterstabilisierung. Differenzierte Behandlungsstrategie mit Suretac, Fastak, Holmium: YAG-Laser und Elektrochirurgie.

The goal for arthroscopic stabilization of anterior glenohumeral instability is to achieve an outcome equivalent to or better than open procedures. A number of arthroscopic procedures have been advocated to reestablish continuity of the inferior glenohumeral ligament complex (IGHLC) with the glenoid. Implantable suture anchors were developed to avoid the problems associated with arthroscopic staple capsulorrhaphy like iatrogenic injury of the glenoid or humeral surface, loosening and migration of the staple. Several transosseous techniques include the need for an accessory posterior incision, the possibility of neurovascular injury (Suprascapular or axillary nerve), and the loosening of the repair after typing over the fascia of the infraspinatus posteriorly. The preferred techniques are cannulated, absorbable fixation device (Suretac) and easy implantable suture anchors made of titanium (Fastak). Even in the hands of experienced arthroscopists, unacceptably high recurrence rates for arthroscopic shoulder stabilization have been reported, due to the steep learning curve for both technical performance and patient selection. Our experience suggests, that if proper selection criteria are employed, normal patients and overhead-athletes may benefit from the advantages of an arthroscopic repair without accepting an increased risk for recurrence. We performed a prospective analysis of 105 shoulders, who underwent arthroscopic stabilization with Suretac or Fastak between 4/96 and 7/98. 48 shoulders were available for followup at least one year. The redislocation rate was 6.25% (3 shoulders) and the rate of subluxation without dislocation also was 6.25%, but none of the shoulders required a second open stabilization. The reason for redislocation or subluxation were 5/6 traumatic injuries, participating in contact sports or in one case a generalized ligamentous laxity. In combination with the LACS-Procedure or the Electro thermally assisted capsular shift (ETACS) not only the capsular detachment but also the capsular redundancy may be adressed and a lower failure rate can be expected.

Arthroscopic shoulder stabilization Differential treatment strategy with Suretac, Fastak, holmium: YAG laser and electrosurgery.

The goal for arthroscopic stabilization of anterior glenohumeral instability is to achieve an outcome equivalent to or better than open procedures. A number of arthroscopic procedures have been advocated to reestablish continuity of the inferior glenohumeral ligament complex (IGHLC) with the glenoid. Implantable suture anchors were developed to avoid the problems associated with arthroscopic staple capsulorrhaphy like iatrogenic injury of the glenoid or humeral surface, loosening and migration of the staple. Several transosseous techniques include the need for an accessory posterior incision, the possibility of neurovascular injury (Suprascapular or axillary nerve), and the loosening of the repair after tying over the fascia of the infraspinatus posteriorly. The preferred techniques are cannulated, absorbable fixation device (Suretac) and easy implantable suture anchors made of titanium (Fastak). Even in the hands of experienced arthroscopists, unacceptably high recurrence rates for arthroscopic shoulder stabilization have been reported, due to the steep learning curve for both technical performance and patient selection. Our experience suggests, that if proper selection criteria are employed, normal patients and overhead-athletes may benefit from the advantages of an arthroscopic repair without accepting an increased risk for recurrence. We performed a prospective analysis of 105 shoulders, who underwent arthroscopic stabilization with Suretac or Fastak between 4/96 and 7/98. 48 shoulders were available for followup at least one year. The redislocation rate was 6.25 % (3 shoulders) and the rate of subluxation without dislocation also was 6.25 %, but none of the shoulders required a second open stabilization. The reason for redislocation or subluxation were 5/6 traumatic injuries, participating in contact sports or in one case a generalized ligamentous laxity. In combination with the LACS-Procedure or the Electro thermally assisted capsular shift (ETACS) not only the capsular detachment but also the capsular redundancy may be adressed and a lower failure rate can be expected.

[Limit values for polycyclic aromatic hydrocarbons in soil of children's playgrounds--basic criteria and recommendations]. / Richtwerte für polyzyklische aromatische Kohlenwasserstoffe im Boden von Kinderspielplätzen--Ableitungskriterien und Empfehlungen.

Elevated concentrations of polycyclic aromatic hydrocarbons (PAK) are often found in the soil of former waste disposal sites, industrial areas, etc. It is desirable and useful to determine orientation values to facilitate and unify the evaluation of contaminations under the aspects of present or planned uses of an area, health protection and decision-making on remedial measures. In the present paper we wish to draw attention to, and discuss problems resulting from, particular characteristics of PAK, e.g. the toxicological property "complete carcinogens" or the necessity of taking into account oral, inhalative and dermal exposure of children on a playground. Based on the discussion, orientation values for benzo[a]pyrene and PAK ("normal" pattern) of 0.5 mg/kg soil and 5 mg/kg soil, respectively, are recommended for top soil of vegetation-free playgrounds. In comparison, deductions carried out by other working groups are presented.

[Evaluation of wood preservative pollutants of indoor air]. / Beurteilung von Holzschutzmittelbelastungen in Innenräumen.

The evaluation of indoor pollution by wood preservatives and pesticides has been a matter of increasing interest for the past years. The present paper reviews actual knowledge. The so-called "wood preservative syndrome" is referred to. Basics and evaluation criteria concerning the investigation of external exposure (wood, dust, air) and human biomonitoring is summarized. Several substances, in particular pentachlorophenol, lindane and pyrethroids, and principle possibilities to decrease exposure are considered.

[Public health evaluation of (hazardous) substances in soil--concept of the AGU and Department of "Tolerable (Hazardous) Substance Concentrations in Soil" for metals]. / Gesundheitliche Bewertung von (Schad-)Stoffen im Boden--Konzept der AGU und Ableitung "Tolerabler (Schad-)Stoffkonzentrationen im Boden" (TSKB) für Metalle.

Up to now no uniform procedure and no generally accepted values exist for health evaluation of pollutants in soil. Aim of the working group for environmental hygiene (AGU) of the Bavarian Ministry of Labour and Social Order, Family, Women and Health is to propose health-based procedures and values for assessment, in consideration of existing concepts and regulations. Basis for health evaluation is the existence of appropriate soil investigations. This refers to the sampling scheme (sampling depth, sampling site and manner, number and representativity of samples) as well as sample preparation and analysis. The "still tolerable pollutant concentration in soil" (TSKB) is supposed to mark a threshold which separates a range of tolerable pollutant concentration from a range where soil-related measures are indicated. Basis for determination of a TSKB-value is the estimation of the average uptake of the respective substance via air, drinking water and food. From this the average level of a tolerable dosis without health risk by regular long-term uptake by humans is calculated. From the remaining portion of tolerable uptake the TSKB-value for the additional pathway of uptake by soil is derived. The estimations are representatively performed for 3-year old children, because small children are especially endangered by uptake of polluted soil. TSKB-values were determined by this procedure for the metals As, Cd, Cr, Cu, Hg, Ni, Pb, Zn and compared with assessment criteria and values of other working groups.

Genotoxicity of 1,3- and 1,6-dinitropyrene: induction of micronuclei in a panel of mammalian test cell lines.

1,3-Dinitropyrene (1,3-DNP) and 1,6-dinitropyrene (1,6-DNP) were assessed for their potential to increase the frequencies of micronuclei in a panel of test cell lines consisting of H4IIEC3/G-, 5L, 5L/r-1,3-DNP1, 208F, V79, V79/r-1,6-DNP1, HepG2 and BWI-J cells, which have been partially characterized for their expression of xenobiotic metabolising enzymes. The micronuclei were analyzed for the presence or absence of kinetochores indicating the occurrence of aneuploidy or chromosome breakage, respectively. 1,3-DNP caused a substantial increase in the frequency of micronuclei only in V79 cells. 1,6-DNP was strongly genotoxic in lines H4IIEC3/G-, 208F, V79 and, to a minor degree, in 5L/r-1,3-DNP1. It caused the formation of kinetochore-positive as well as kinetochore-negative micronuclei in V79 cells but only of kinetochore-negative micronuclei in H4IIEC3/G- and 208F cells. 1,6-DNP-induced formation of micronuclei was paralleled by the appearance of multinucleated cells. Treatment of V79 cells with 1,3-DNP resulted in the same types of damage as treatment with 1,6-DNP, although considerably higher concentrations were required. The results show that 1,6-DNP can be highly genotoxic in mammalian cells, whereas, at least in the panel of test cell lines used, 1,3-DNP possesses only a low genotoxic activity. 1,3-DNP appears to be activated to genotoxic products in V79 cells by the same pathway(s) as 1,6-DNP.

Induction of DNA damage by N-nitrosodiethylamine in rat hepatoma cells: correlation with cytochrome P450-mediated aldrin epoxidase activity.

The genotoxic potency of the hepatocarcinogen N-nitrosodiethylamine (NDEA) was tested in various Reuber hepatoma cell lines, which differ in their expression of differentiated liver specific functions including hepatic cytochrome P450 forms, and in rat hepatocytes, with the aim of characterizing the enzymes involved in activation. DNA single-strand breaks assessed by alkaline elution served as an indicator of genetic damage. Aldrin epoxidase activity was used as a marker for various hepatic cytochrome P450 forms. The poorly differentiated cell lines RH35 and H4IIEC3/T were apparently not affected by NDEA; moderate effects were observed in the well-differentiated lines H4IIEC3/G- and 2sFou, and major effects in two other well-differentiated lines, Fao and C2Rev7, and in hepatocytes. The degree of DNA damage in the cell lines correlated positively with the expression of aldrin epoxidase. Furthermore, DNA damage induced by N-nitrosodimethylamine (NDMA) was determined in C2Rev7 cells and in rat hepatocytes in order to assess a possible involvement of the NDMA-metabolizing cytochrome P45oIIE1 in the activation of NDEA by comparing the genotoxic potencies of the two compounds. NDMA was distinctly less effective than NDEA in C2Rev7 cells at all concentrations tested. In hepatocytes, NDMA induced more DNA damage than NDEA at low concentrations, but was slightly less active at high concentrations. The results suggest that NDEA is preferentially metabolized to genotoxic products by one or several cytochrome P450 forms different from P450IIE1.

Involvement of different pathways in the genotoxicity of nitropropanes in cultured mammalian cells.

The metabolic pathways leading to genotoxicity of nitropropanes in mammalian cells were investigated by measuring the effects of 2-nitropropane (2-NP) and 1-nitropropane (1-NP) on various cell lines characterized for their expression of cytochrome P450-dependent mono-oxygenases. Cells used were the rat hepatoma cell lines 2sFou, H4IIEC3/G- and C2Rev7, which express various forms of cytochrome P450-dependent mono-oxygenases, and V79 Chinese hamster cells which lack these enzyme activities. Induction of DNA repair synthesis, micronuclei and, where assessable, mutations to 6-thioguanine (TG) resistance served as indicators of genotoxic effects. 2-NP elicited a positive response at all endpoints measured in the hepatoma lines after pretreatment of the cells with dexamethasone, an inducer of various liver-specific cytochrome P450 forms. Genotoxicity was much weaker or not detectable in cells not pretreated with the inducer. 1-NP was not genotoxic in the hepatoma cells irrespective of whether the cells were pretreated or not. Neither isomer elicited DNA repair synthesis in V79 cells, but both isomers caused mutations to TG resistance, and 1-NP increased the number of micronucleated and multinucleated cells. The findings show that there are different pathways in mammalian cells by which nitropropanes can be converted to genotoxic products. Presumably the induction of liver tumours by 2-NP is linked to the metabolic pathway which is characterized by the formation of genotoxic metabolites from 2-NP but not 1-NP. This pathway appears to depend on the presence of liver-specific, dexamethasone-inducible, cytochrome P450 forms. The relevance of the genotoxic effects of the nitropropanes observed in V79 cells for the situation in vivo is open to question.

H4IIEC3 rat hepatoma cells activate N-nitrosodimethylamine but are resistant to the genotoxic products.

Exposure of the differentiated rat hepatoma cells H4IIEC3 (H4) or several of their subclones to 20 mM N-nitrosodimethylamine (NDMA) did not significantly increase the number of 6-thioguanine (TG)-resistant cells or of micronuclei. Similar results were obtained with H4 cells pretreated with isopropanol, an inducer of the NDMA-metabolizing cytochrome P450 form. However, when H4 cells were co-cultured with V79 Chinese hamster cells, which are incapable of activating NDMA, exposure to the nitrosamine (5-40 mM) caused a concentration-dependent increase in the frequency of TG-resistant V79 cells; pretreatment of H4 cells with 0.5% isopropanol nearly doubled the magnitude of this response. When freshly isolated rat hepatocytes were used as the activation system in co-cultures with either H4 cells or V79 cells, NDMA induced up to 20 times more TG-resistant mutants in V79 cells than in H4 cells. The results indicate that H4 hepatoma cells are capable of metabolizing NDMA to genotoxic products but can protect themselves, presumably by repairing the potentially mutagenic DNA lesions.

Ski-flying: related catecholamine excretion compared with cross-country skiing.

We examined the catecholamine excretion and its performance-diagnostic relevance in athletes of the German National Team during ski-flying and cross-country skiing. Five athletes were examined during the 1986 World Championship ski-flying competition and eight athletes during a 24-km cross-country qualification test. There is little doubt that the special competitive conditions have considerable influence on the usefulness of these data. We decided to publish the results anyway since the results may expand our knowledge about the performance-diagnostic relevance of catecholamine excretion under competitive conditions. During cross-country skiing, the average noradrenaline elimination (1166 pmol.min-1) was about 150% higher and the average adrenaline elimination (243 pmol.min-1) about 30% lower than during ski-flying. The noradrenaline-adrenaline ratio was about 4.8 in cross-country skiing and 1.3-1.5 in ski-flying. Catecholamine excretion does, as expected, therefore permit differentiation of the various strains on the organism. The impression with respect to performance-diagnostic relevance arises that successful athletes show a more stable and lower excretion of noradrenaline under competitive conditions. This can only be a preliminary statement; it should serve to stimulate discussion and further investigations of the problem.

1,6-Dinitropyrene causes spindle disturbances and chromosomal damage in V79 Chinese hamster cells.

We have investigated the cytogenetic effect of 1,6-dinitropyrene (1,6-DNP) in Chinese hamster V79 cells. The chemical caused a dose-dependent increase in the incidence of initial and full C-mitoses, polyploid mitoses, ana-telophases with lagging chromosomes, non-disjunction and multipolar configurations, in a range of 0.05-5 microM. These findings indicate that 1,6-DNP interferes with the functioning of the spindle apparatus in V79 cells. Early signs of spindle disturbances were seen at 1,6-DNP concentrations which only moderately reduced cell growth and division. Analysis of structural chromosomal aberrations revealed the appearance of chromatid-type aberrations with open breaks and exchanges accompanied by gaps. The results indicate that 1,6-DNP is both a spindle-disturbing and a clastogenic agent in V79 cells.

Mutagenicity, clastogenicity and cytotoxicity of procarcinogens in a rat hepatoma cell line competent for xenobiotic metabolism.

The present studies were aimed at evaluating the suitability of the differentiated Reuber hepatoma cells H4IIEC3/G- for monitoring permanent damage to the DNA caused by hepatotrophic chemicals. First we determined the profile of xenobiotic metabolizing enzymes. The cells expressed various cytochrome P-450-dependent monooxygenases, UDP-glucuronosyl-, phenol sulpho- and glutathione S-transferase, cytochrome c (P-450) reductase and carboxylesterases. We then established the conditions for genotoxicity testing in H4IIEC/G- cells. Induction of resistance against 6-thioguanine and appearance of micronuclei served as indicators for mutagenicity and clastogenicity, respectively. 6-Thioguanine-resistant H4IIEC3/G- cells were phenotypically stable for at least 30 cell cycles; recovery of 6-thioguanine-resistant cells was not significantly affected by the number of cells seeded for mutant selection up to at least 10(6) cells/100-mm dish; expression time of chemically induced mutants was 12-15 days; a period of 24 h after treatment appeared to be sufficient to allow for the formation of micronuclei. Finally we tested the genotoxic effects of promutagens which are typically activated or inactivated in liver. Aflatoxin B1, N-nitrosodiethylamine and cyclophosphamide were genotoxic to H4IIEC3/G- cells at concentrations of 10-30 nM, 2-20 mM and 1 mM, respectively. N-Nitrosodimethylamine and benzo[a]pyrene were not or only weakly cytotoxic and genotoxic to the cells, but this appears most likely to be due to protective mechanisms rather than to lack of metabolic activation. The results indicate that differentiated hepatoma cells such as H4IIEC3/G- offer a means of studying the potential of chemicals for inducing permanent DNA damage in liver cells.

The effects of light quality and intensity on the synthesis of ribulose-1,5-bisphosphate carboxylase and its mRNAs in the green alga Chlorogonium elongatum.

In the green alga Chlorogonium elongatum the promoting effect of light on the synthesis of ribulose bisphosphate carboxylase/oxygenase (RuBPCase) is mainly caused by blue light of wavelengths between 430 nm and 510 nm, with a maximum effect at about 460 nm. Blue light also causes an increase in the amounts of the mRNAs for the large and the small subunits of the enzyme. Furthermore, the concentration of RuBPCase is affected by the light energy fluence rate. The rate of synthesis as well as the maximal obtainable concentration of the enzyme are functions of the light energy fluence rate up to 26 W·m(-2). No further increase occurs beyond that intensity. The quantity of irradiation also alters the concentrations of the subunit mRNAs. The results indicate that the changes in the mRNA levels are the major regulatory steps in the light-dependent synthesis of the RuBPCase enzyme.