Correction: Degen et al. Citrinin Exposure in Germany: Urine Biomarker Analysis in Children and Adults. Toxins 2023, 15, 26.

In the original publication [...].

Human duodenal organoid-derived monolayers serve as a suitable barrier model for duodenal tissue.

Usually, duodenal barriers are investigated using intestinal cell lines like Caco-2, which in contrast to native tissue are limited in cell-type representation. Organoids can consist of all intestinal cell types and are supposed to better reflect the in vivo situation. Growing three-dimensionally, with the apical side facing the lumen, application of typical physiological techniques to analyze the barrier is difficult. Organoid-derived monolayers (ODMs) were developed to overcome this. After optimizing culturing conditions, ODMs were characterized and compared to Caco-2 and duodenal tissue. Tight junction composition and appearance were analyzed, and electrophysiological barrier properties, like paracellular and transcellular barrier function and macromolecule permeability, were evaluated. Furthermore, transcriptomic data were analyzed. ODMs had tight junction protein expression and paracellular barrier properties much more resembling the originating tissue than Caco-2. Transcellular barrier was similar between ODMs and native tissue but was increased in Caco-2. Transcriptomic data showed that Caco-2 expressed fewer solute carriers than ODMs and native tissue. In conclusion, while Caco-2 cells differ mostly in transcellular properties, ODMs reflect trans- and paracellular properties of the originating tissue. If cultured under optimized conditions, ODMs possess reproducible functionality, and the variety of different cell types makes them a suitable model for human tissue-specific investigations.

Comparison of different salt solutions for density separation of conventional and biodegradable microplastic from solid sample matrices.

Microplastics are the new emerging pollutants ubiquitously detectable in aquatic and terrestrial ecosystems. Fate and behavior, as well as ecotoxicity, are of increasing environmental concern, particularly in sediments and soils as natural sinks. For a global environmental risk assessment, reliable and easy to apply analytical methods are mandatory to obtain comparable data. This is based on the isolation of microplastics out of the solid sample matrices prior to instrumental detection. Thus, this study provides an easy to apply approach for density separation. The technique emerged from a comparative study using different salt solutions to isolate conventional, and for the first time biodegradable, microplastics from different solid sample matrices, i.e., sand, artificial soil, and compost. Four solutions (water, sodium chloride, sodium hexametaphosphate, and sodium bromide) of different densities were applied followed by oxidizing digestion. Finally, the impact of the procedures on size and surface properties of microplastics was tested. Dependent on the sample matrix, the highest recovery rates of 87.3-100.3% for conventional polymers, and 38.2-78.2% for biodegradable polymers, were determined with sodium bromide. It could be shown that the type of solid sample matrix influences the recovery rates and has to be considered when choosing a sample preparation technique.

Dissection of Barrier Dysfunction in Organoid-Derived Human Intestinal Epithelia Induced by Giardia duodenalis.

BACKGROUND & AIMS: The protozoa Giardia duodenalis is a major cause of gastrointestinal illness worldwide, but underlying pathophysiological mechanisms remain obscure, partly due to the absence of adequate cellular models. We aimed at overcoming these limitations and recapitulating the authentic series of pathogenic events in the primary human duodenal tissue by using the human organoid system. METHODS: We established a compartmentalized cellular transwell system with electrophysiological and barrier properties akin to duodenal mucosa and dissected the events leading to G. duodenalis-induced barrier breakdown by functional analysis of transcriptional, electrophysiological, and tight junction components. RESULTS: Organoid-derived cell layers of different donors showed a time- and parasite load-dependent leak flux indicated by collapse of the epithelial barrier upon G. duodenalis infection. Gene set enrichment analysis suggested major expression changes, including gene sets contributing to ion transport and tight junction structure. Solute carrier family 12 member 2 and cystic fibrosis transmembrane conductance regulator-dependent chloride secretion was reduced early after infection, while changes in the tight junction composition, localization, and structural organization occurred later as revealed by immunofluorescence analysis and freeze fracture electron microscopy. Functionally, barrier loss was linked to the adenosine 3',5'-cyclic monophosphate (cAMP)/protein kinase A-cAMP response element-binding protein signaling pathway. CONCLUSIONS: Data suggest a previously unknown sequence of events culminating in intestinal barrier dysfunction upon G. duodenalis infection during which alterations of cellular ion transport were followed by breakdown of the tight junctional complex and loss of epithelial integrity, events involving a cAMP/protein kinase A-cAMP response element-binding protein mechanism. These findings and the newly established organoid-derived model to study G. duodenalis infection may help to explore new options for intervening with disease and infection, in particular relevant for chronic cases of giardiasis.

Citrinin Exposure in Germany: Urine Biomarker Analysis in Children and Adults.

Citrinin (CIT), a mycotoxin known to exert nephrotoxicity, is a contaminant in food and feed. Since CIT contamination is not regularly analyzed, data on its occurrence and especially levels in food commodities are insufficient for conducting a conventional exposure assessment. Yet, human biomonitoring, i.e., an analysis of CIT and its metabolite dihydrocitrinone (DH-CIT) in urine samples allows to estimate exposure. This study investigated CIT exposure in young (2-14 years) and adult (24-61 years) residents of three federal states in Germany. A total of 179 urine samples from children and 142 from adults were collected and analyzed by a targeted LC-MS/MS based method for presence of CIT and DH-CIT. At least one of the biomarkers was detected and quantified in all urines, which indicated a widespread dietary exposure to the mycotoxin in Germany. Interestingly, the biomarker concentrations of CITtotal (sum of CIT and DH-CIT) were higher in children's urine (range 0.05-7.62 ng/mL; median of 0.54 ng/mL) than in urines from adults (range 0.04-3.5 ng/mL; median 0.3 ng/mL). The biomarker levels (CITtotal) of individual urines served to calculate the probable daily CIT intake, for comparison to a value of 0.2 µg/kg bw/day defined as 'level of no concern for nephrotoxicity' by the European Food Safety Authority. The median exposure of German adults was 0.013 µg/kg b.w., with only one urine donor exceeding this provisional tolerable daily intake (pTDI) for CIT. The median exposure of children was 0.05 µg/kg bw per day (i.e., 25% of the pTDI); however, CIT exposure in 12 individuals (6.3% of our study group) exceeded the limit value, with a maximum intake of 0.46 µg/kg b.w. per day. In conclusion, these results show evidence for non-negligible exposure to CIT in some individuals in Germany, mainly in children. Therefore, further biomonitoring studies and investigations aimed to identify the major sources of CIT exposure in food commodities are required.

A Comparison of Different Counting Methods for a Holographic Particle Counter: Designs, Validations and Results.

DIH is used in many fields of 3D imaging to locate micro or nano-particles in a volume and determine their size, shape or trajectories. A variety of different wavefront reconstruction approaches have been developed for 3D profiling and tracking to study particles' morphology or visualize flow fields. The novel application of HPCs requires observing particle densities in a given sampling volume which does not strictly necessitate the reconstruction of particles. Such typically spherical objects yield circular intereference patterns-also referred to as fringe patterns-at the hologram plane which can be detected by simpler 2D image processing means. The determination of particle number concentrations (number of particles/unit volume [#/cm 3 ]) may therefore be based on the counting of fringe patterns at the hologram plane. In this work, we explain the nature of fringe patterns and extract the most relevant features provided at the hologram plane. The features aid the identification and selection of suitable pattern recognition techniques and its parameterization. We then present three different techniques which are customized for the detection and counting of fringe patterns and compare them in terms of detection performance and computational speed.

Design and Validation of a Holographic Particle Counter.

An in-line holographic particle counter concept is presented and validated where multiple micrometer sized particles are detected in a three dimensional sampling volume, all at once. The proposed PIU is capable of detecting holograms of particles which sizes are in the lower µ m- range. The detection and counting principle is based on common image processing techniques using a customized HT with a result directly relating to the particle number concentration in the recorded sampling volume. The proposed counting unit is mounted ontop of a CNM for comparison with a commercial TSI-3775 CPC. The concept does not only allow for a precise in-situ determination of low particle number concentrations but also enables easy upscaling to higher particle densities (e.g., > 30 . 000 # c c m ) through its linear expandability and option of cascading. The impact of coincidence at higher particle densities is shown and two coincidence correction approaches are presented where, at last, its analogy to the coincidence correction methods used in state-of-the-art CPCs is identified.

Characterization of a Robust 3D- and Inkjet-Printed Capacitive Position Sensor for a Spectrometer Application.

An inkjet- and 3D-printed capacitive sensor system with an all-digital and flexible sensor read-out hardware is reported. It enables spectrometer devices with significantly reduced device outlines and costs. The sensor is developed as multilayer inkjet-printed electrode structure on a 3D-printed copper housing. Very high required position resolutions of r e s p o s < 50 nm and a wide measurement range of r m = 1000 µ m at an offset of d 0 = 1000 µ m in the considered spectrometers motivate this work. The read-out hardware provides high sampling rates of up to r s ≈ 10 ns and enables the generation of trigger signals, i.e., the mirror control signal, without a time lag. The read-out circuitry is designed as a carrier frequency system, which enables flexible choices of bandwidth and measurement signal frequency. It thus allows for separation in frequency from coupling parasitics, i.e., other frequencies present in the device under test, and makes the read-out quasi-noise-immune.

Pain extent is more strongly associated with disability, psychological factors, and neck muscle function in people with non-traumatic versus traumatic chronic neck pain: a cross sectional study.

BACKGROUND: Neck pain is presented with a variety of symptoms. Pain drawings are used in the clinical assessment of people with neck pain. Pain extent based upon pain drawings can be associated with different factors. However, the relation between pain extent and function limitations in people with neck pain is unknown. AIM: The aim of this study was to explore the associations between pain extent extracted from pain drawings, and self-reported neck pain related disability, quality of life, depression, self-reported neck function, cervical muscle function, and range of motion in a chronic neck pain population and possible differences depending on the onset of pain being traumatic or not. DESIGN: Observational cross-sectional study. SETTING: Primary and secondary healthcare. POPULATION: People with chronic neck pain (N.=200) of traumatic (N.=120) or non-traumatic (N.=80) origin. METHODS: Outcome measures: Pain extent, Short Form 36 Health Survey Physical and Mental Component Summary (SF36-PCS/MCS), TAMPA Scale of Kinesiophobia (TSK), Beck Depression Inventory-II (BDI-II), Neck Disability Index (NDI), Craniocervical Flexion Test (CCFT), Cervical Extension Test (CE), and Cervical Range of Motion (ROM). Correlations were calculated using Spearman or Pearson correlation coefficients. Correlation between pain extent and outcomes were calculated for all participants collectively and then separately for those with a traumatic versus non-traumatic neck pain. RESULTS: Overall, significant positive correlations were observed between pain extent and NDI (r=0.33; P<0.001), BDI-II (r=0.29; P<0.001), CCFT (r=-0.24; P=0.001) and CE (r=-0.19; P=0.006). No difference was observed in pain extent between patients with traumatic (mean: 7.6±6.7%) and non-traumatic onset (7.4±6.8%). Pain extent correlated moderately with NDI, BDI-II, TSK, CCFT and CE in those with non-traumatic onset, but weakly with NDI, BDI-II, CCFT and CE in those with trauma-induced chronic neck pain. CONCLUSIONS: Pain extent is correlated with patient-reported neck function, depression and muscle test performance in people with chronic neck pain. These correlations were strongest in those with non-traumatic neck pain. CLINICAL REHABILITATION IMPACT: Patients presenting with larger pain areas show poorer psychological and physical function. Pain drawings can therefore indicate a need for addressing these functions in management of a person with chronic neck pain.

Occurrence of carbazoles in dust and air samples from different locations in Germany.

9H-carbazole is generated from incomplete combustion of diverse fossil fuels and biomass, in tobacco smoke and from industrial processes, while halogenated carbazoles have natural and anthropogenic sources. We analyzed 9H-carbazole and 14 halogenated carbazoles in dust samples from 14 schools, 13 daycare centers, and 13 residences, as well as 5 indoor air samples from residences in Munich, Germany. Overall, we present first data of various carbazoles in different indoor environments without visible combustion sources. The median (95th percentile) values of the halogenated analytes mainly detected in the entire study group were 10.3ng/g (308ng/g) for 9H-carbazole, 13.3ng/g (735ng/g) for 3,6-dichloro-9H-carbazole, 6.2ng/g (159ng/g) for 1,3,6-tribromo-9H-carbazole, and 1.2ng/g (21.1ng/g) for 2,7-dibromo-9H-carbazole. For most of the target analytes, the highest concentrations were observed in dust samples from schools, and the lowest were found in residences. In the air samples, all analytes were found only at low levels, with median values of 7.7pg/m3 for 9H-carbazole and 6.1pg/m3 for 2,3,6,7-tetrachloro-9H-carbazole. For 9H-carbazole, "typical" and "high" non-dietary intake of children through dust ingestion using median and 95th percentile values were calculated to be 0.03ng/kg b.w. and 1.1ng/kg b.w. daily, respectively. Due to limited toxicological information and exposure data for other relevant pathways (e.g., dietary intake), the risk assessment is inconclusive. Nevertheless, there are indications that 9H-carbazole has carcinogenic properties and that halogenated carbazoles have dioxin-like toxicities. Therefore, further research is essential.

Giardia's Epithelial Cell Interaction In Vitro: Mimicking Asymptomatic Infection?

The protozoan parasite Giardia duodenalis is responsible for more than 280 million cases of gastrointestinal complaints ("giardiasis") every year, worldwide. Infections are acquired orally, mostly via uptake of cysts in contaminated drinking water. After transformation into the trophozoite stage, parasites start to colonize the duodenum and upper jejunum where they attach to the intestinal epithelium and replicate vegetatively. Outcome of Giardia infections vary between individuals, from self-limiting to chronic, and asymptomatic to severely symptomatic infection, with unspecific gastrointestinal complaints. One proposed mechanism for pathogenesis is the breakdown of intestinal barrier function. This has been studied by analyzing trans-epithelial electric resistances (TEER) or by indicators of epithelial permeability using labeled sugar compounds in in vitro cell culture systems, mouse models or human biopsies and epidemiological studies. Here, we discuss the results obtained mainly with epithelial cell models to highlight contradictory findings. We relate published studies to our own findings that suggest a lack of barrier compromising activities of recent G. duodenalis isolates of assemblage A, B, and E in a Caco-2 model system. We propose that this epithelial cell model be viewed as mimicking asymptomatic infection. This view will likely lead to a more informative use of the model if emphasis is shifted from aiming to identify Giardia virulence factors to defining non-parasite factors that arguably appear to be more decisive for disease.

Alkylsulfonic acid phenylesters (ASEs, Mesamoll®) in dust samples of German residences and daycare centers (LUPE 3).

For decades, plasticizers have been produced in high quantities to improve the flexibility and durability of products. One possible replacement product is alkylsulfonic acid phenylesters (ASEs), marketed as Mesamoll®. This study aimed to quantify the ASE dust contamination of residences and daycare centers to obtain insight into the recent exposure situation. ASEs were quantified in dust samples collected from 25 residences and 25 daycare centers using GC/MS measurements. Median (95th percentile) values of the sum of tetra- to heptadecylphenylesters are higher in daycare centers, with a value of 19.6mg/kg (216mg/kg), compared to residences, with a value of 7.6mg/kg (171mg/kg). A daily non-dietary intake of 0.08 and 0.86µg/kg b.w., respectively, was observed using the median and 95th percentile values obtained from dust samples. These levels are 1250 and 115 times below a previously set temporary tolerable daily intake value. Nevertheless, the fact that basic data on toxicity and exposure via other pathways are limited or unavailable at present has to be considered.

Monocyclic and bicyclic monoterpenes in air of German daycare centers and human biomonitoring in visiting children, the LUPE 3 study.

To investigate the assumed association between indoor air pollution with monoterpenes (MTps) and the internal MTp exposure of occupants, a comparative study was performed in daycare centers in two federal states of Germany. Three well-known monoterpenoid air pollutants, viz. α-pinene (αPN), Δ(3)-carene (CRN), and R-limonene (LMN), were measured in indoor air in 45 daycare centers. Additionally, urine samples of 222 children visiting these facilities were collected in the evening after a full-day stay. Altogether 11 MTp metabolites were analyzed in the urine samples using a novel highly sensitive and selective gas chromatographic-tandem-mass spectrometric procedure. The medians (95th percentiles) of the MTp levels in indoor air were 9.1 µg m(-3) (94 µg m(-3)) for LMN, 2.6 µg m(-3) (13 µg m(-3)) for αPN, and <1.0 µg m(-3) (3.2 µg m(-3)) for CRN. None of the day care centers exceeded the German health precaution or hazard guide value. In spite of the low MTp air exposure, the urine analyses revealed an exposure to the three monoterpenes in almost all children. The median levels of MTp metabolites in urine were 0.11 mg L(-1) for LMN-8,9-OH, 0.10 mg L(-1) for LMN-1,2-OH, 49 µg L(-1) for PA, 2.9 µg L(-1) for POH, 5.2 µg L(-1) for tCAR, and 4.1 µg L(-1) for cCAR (LMN metabolites), 7.2 µg L(-1) for MYR, 19 µg L(-1) for tVER, and 19 µg L(-1) for cVER (αPN metabolites), as well as 8.2 µg L(-1) for CRN-10-COOH (CRN metabolite). Statistically significant and strong correlations among the urinary metabolites of each MTp were found. Moreover, statistical associations between LMN metabolites and the LMN indoor air levels were revealed. However, the weakness of the associations indicates a considerable impact of other MTp sources, e.g. diet and consumer products, on the internal exposure.

Studying enzymatic bioreactions in a millisecond microfluidic flow mixer.

In this study, the pre-steady state development of enzymatic bioreactions using a microfluidic mixer is presented. To follow such reactions fast mixing of reagents (enzyme and substrate) is crucial. By using a highly efficient passive micromixer based on multilaminar flow, mixing times in the low millisecond range are reached. Four lamination layers in a shallow channel reduce the diffusion lengths to a few micrometers only, enabling very fast mixing. This was proven by confocal fluorescence measurements in the channel's cross sectional area. Adjusting the overall flow rate in the 200 µm wide and 900 µm long mixing and observation channel makes it possible to investigate enzyme reactions over several seconds. Further, the device enables changing the enzyme/substrate ratio from 1:1 up to 3:1, while still providing high mixing efficiency, as shown for the enzymatic hydrolysis using ß-galactosidase. This way, the early kinetics of the enzyme reaction at multiple enzyme/substrate concentrations can be collected in a very short time (minutes). The fast and easy handling of the mixing device makes it a very powerful and convenient instrument for millisecond temporal analysis of bioreactions.

Perfluorinated compounds in fish and blood of anglers at Lake Möhne, Sauerland area, Germany.

Perfluorinated compounds (PFCs) were measured in fish samples and blood plasma of anglers in a cross-sectional study at Lake Möhne, Sauerland area, Germany. Human plasma and drinking water samples were analyzed by solid phase extraction, high-performance liquid chromatography (HPLC), and tandem mass spectrometry (MS/MS). PFCs in fish fillet were measured by ion pair extraction followed by HPLC and MS/MS. PFOS concentrations in 44 fish samples of Lake Möhne ranged between 4.5 and 150 ng/g. The highest median PFOS concentrations have been observed in perches (median: 96 ng/g) and eels (77 ng/g), followed by pikes (37 ng/g), whitefish (34 ng/g), and roaches (6.1 ng/g). In contrast, in a food surveillance program only 11% of fishes at retail sale contained PFOS at detectable concentrations. One hundred five anglers (99 men, 6 women; 14-88 years old; median 50.6 years) participated in the human biomonitoring study. PFOS concentrations in blood plasma ranged from 1.1 to 650 µg/L (PFOA: 2.1-170 µg/L; PFHxS: 0.4-17 µg/L; LOD: 0.1 µg/L). A distinct dose-dependent relationship between fish consumption and internal exposure to PFOS was observed. PFOS concentrations in blood plasma of anglers consuming fish 2-3 times per month were 7 times higher compared to those without any fish consumption from Lake Möhne. The study results strongly suggest that human internal exposure to PFC is distinctly increased by consumption of fish from PFC-contaminated sites.

Time-resolved mid-IR spectroscopy of (bio)chemical reactions in solution utilizing a new generation of continuous-flow micro-mixers.

A specially designed micro-mixer made of silicon, calcium fluoride, and silicone with an optical transmission path of 8 µm has been used for mid-IR spectroscopy monitoring of mixing-induced chemical reactions in the low millisecond time regime. The basic principle of the proposed continuous-flow technique is to mix two liquids introduced in two times two alternatingly stacked layers through diffusion at the entrance of a 200 µm wide, 1 cm long micro-fluidic channel also serving as measurement area. By using this special, dedicated arrangement, diffusion lengths and hence the mixing times can be significantly shortened and the overall performance improved in comparison to previous systems and alternative methods. Measurements were carried out in transmission mode using an Fourier transform infrared (FTIR) microscope, recording spectra with spot sizes of 180 × 100 µm(2) each at defined spots along this channel. Each of these spots corresponds to a specific reaction time: moving the measurement spot towards the entry yields shorter reaction times, moving it towards the channel's end gives longer reaction times. This principle is generic in nature and provides a solution for accurate, chemically induced triggering of reactions requiring the mixing of two liquid reagents or reagent solutions. A typical experiment thus yields up to 85 time-coded data points, covering a time span from 1 to 80 ms at a total reagent consumption of only about 125 µL. Using the fast neutralization reaction of acetic acid with sodium hydroxide as a model, the time required for 90% mixing was determined to be around 4 ms. Additionally, first experiments on ubiquitin changing its secondary structure from native to "A-state" were carried out, illustrating the potential for time-resolved measurements of proteins in aqueous solutions.

Expression of the human endogenous retrovirus (HERV) group HML-2/HERV-K does not depend on canonical promoter elements but is regulated by transcription factors Sp1 and Sp3.

After fixation in the human genome, human endogenous retroviruses (HERVs) are bona fide cellular genes despite their exogenous origin. To be able to spread within the germ line and the early embryo, the ancient retroviral promoters must have adapted to the requirements for expression in these cell types. We describe that in contrast to the case for current exogenous retroviruses, which replicate in specific somatic cells, the long terminal repeat (LTR) of the human endogenous retrovirus HERV-K acts as a TATA- and initiator element-independent promoter with a variable transcription start site. We present evidence that the HERV-K LTR is regulated by the transcription factors Sp1 and Sp3. Mutating specific GC boxes, which are binding sites for Sp proteins, and knocking down Sp1 and Sp3 by use of small interfering RNA (siRNA) significantly reduced the promoter activity. Binding of Sp1 and Sp3 to the promoter region was confirmed using electrophoretic mobility shift assays (EMSAs) and chromatin immunoprecipitation (ChIP). Our data explain why certain HERV-K proviruses have lost promoter competence. Since vertebrate promoters lacking canonical core promoter elements are common but poorly studied, understanding the HERV-K promoter not only will provide insight into the regulation of endogenous retroviruses but also can serve as a paradigm for understanding the regulation of this class of cellular genes.

Prevalence of nickel sensitization and urinary nickel content of children are increased by nickel in ambient air.

In a cross-sectional study performed in 2000, an unexpected positive association between nickel (Ni) in ambient air, urinary Ni content and the prevalence of Ni sensitization in a subgroup of 6-yr-old children living near a steel mill was observed. Between 2005 and 2006, in a different and larger study population, we examined if Ni from ambient air or urinary Ni concentration was related to Ni sensitization in children living next to Ni-emitting steel mills. We studied 749 school beginners living in four Ni-polluted industrial areas of North Rhine-Westphalia, Germany. We assessed Ni in ambient air, Ni in urine from children and mothers, and Ni in tap water, conducted patch tests in children (including the NiSO(4)-dilution test) and collected questionnaire data. Statistics were done by linear and logistic regression analyses, adjusted for covariates. At increased Ni concentration in ambient air (unit of increase: 10 ng/m(3)), urinary Ni concentrations rose in both mothers (9.1%; 95% CI: 6.8-11.4%) and children (2.4%; 95% CI: 0.4-4.4%). The prevalence of Ni sensitization in children was associated with increased Ni from ambient air (unit of increase: 18 ng/m(3); odds ratio 1.28; 95% CI: 1.25-1.32) and urinary Ni concentration (unit of increase: 7.1 µg/L; odds ratio 2.4; 95% CI: 1.20-4.48). Ni in ambient air of areas with Ni-emitting factories contributes to internal Ni exposure in residents via inhalation and, furthermore, is a risk factor for the development of Ni sensitization in children.

Two-year follow-up biomonitoring pilot study of residents' and controls' PFC plasma levels after PFOA reduction in public water system in Arnsberg, Germany.

Residents in Arnsberg, Germany, had been supplied by drinking water contaminated with perfluorooctanoate (PFOA). Biomonitoring data from 2006 evidenced that plasma PFOA concentrations of residents from Arnsberg were 4.5-8.3 times higher than those in reference groups. The introduction of charcoal filtration in July 2006 distinctly reduced PFOA concentrations in drinking water. Our one-year follow-up study showed a 10-20% reduction of PFOA plasma levels in residents from Arnsberg. Here we report the first results of the two-year follow-up study Arnsberg 2008. Additionally, the results of the two-year follow-up examination of the reference group are included. Paired plasma samples of 138 study participants (45 children, 46 mothers and 47 men) collected in 2006 and 2008 were considered in the statistical analyses. Within the two years plasma concentrations of PFOA, perfluorooctanesulfonate (PFOS) and perfluorohexanesulfonate (PFHxS) decreased in residents from Arnsberg and in control groups. The geometric means of PFOA plasma levels declined by 39% (children and mothers) and 26% (men) in Arnsberg and by 13-15% in the corresponding subgroups from the reference areas. For the population from Arnsberg a geometric mean plasma PFOA half-life of 3.26 years (range 1.03-14.67 years) was calculated. Our results confirm an ongoing reduction of the PFOA load in residents from Arnsberg. The decline of PFC levels in plasma of participants from the reference areas reflects the general decrease of human PFC exposure during the very recent years.