Reconstruction of phthalate exposure and DINCH metabolites from biomonitoring data from the EXHES cohort of Tarragona, Spain: A case study on estimated vs reconstructed DEHP using the PBPK model.

Phthalates are known endocrine disruptors (EDs) and are associated with potential diseases, such as obesity and diabetes. In 2002, the plasticizer 1,2-cyclohexane dicarboxylic acid diisononyl ester (DINCH) was introduced as an alternative to phthalates in the European market. The objective of this study was to evaluate the total exposure to phthalate and DINCH metabolites from EXHES Tarragona, Spain cohort of pregnant women. On the one hand, the analytical determination of phthalate and DINCH metabolites in urine was carried out. On the other hand, the reconstructed exposure was calculated for phthalates and DINCH using their metabolites concentration measured in the urine. Thirteen different phthalate metabolites and two metabolites of DINCH were measured and detected in almost all pregnant women's urine samples (n = 60). There were significant correlations between metabolites of the same parent compounds, and also between DEHP and MBzP metabolites, DiNP and BBZP metabolites, and DEHP and DiNP metabolites respectively. The exposure of pregnant women to phthalate and DINCH parent compounds were also back calculated using the levels of each metabolite found in pregnant women urine (reconstructed exposure). Besides, to demonstrate the utility of this approach, the physiologically based pharmacokinetic (PBPK) model was used to predict the cumulative amount of MEHP (a principal metabolite of DEHP in urine). To proceed with that, DEHP reconstructed exposure and estimated exposure from the same cohort (previously studied by the same authors) were simulated using the PBPK model. Results showed that the reconstructed-PBPK simulation was closer to the 24 h biomonitoring data than the estimated PBPK-simulation., This clearly shows that the combination of reconstructed exposure with the PBPK model is a good tool to predict chemicals exposure. However, some discrepancies between simulated and biomonitored values were found. This can be associated with other sources that contribute to the total exposure and emphasises the need to consider multi-routes exposure for the widely distributed chemicals like phthalates and DINCH.

Comparing dietary and non-dietary source contribution of BPA and DEHP to prenatal exposure: A Catalonia (Spain) case study.

Bisphenol A (BPA) and Di-(2-ethylhexyl) phthalate (DEHP) are two wide spread chemicals classified as endocrine disruptors (ED). The present study aims to estimate the non-dietary (dermal, non-dietary ingestion and inhalation) exposure to BPA and DEHP for a pregnant women cohort. In addition, to assess the prenatal exposure for the fetus, a physiologically based pharmacokinetic (PBPK) model was used. It was adapted for pregnancy in order to assess the internal dosimetry levels of EDs (BPA and DEHP) in the fetus. Estimates of exposure to BPA and DEHP from all pathways along with their relative importance were provided in order to establish which proportion of the total exposure came from diet and which came from non-dietary exposures. In this study, the different oral dosing scenarios (dietary and non-dietary) were considered keeping inhalation as a continuous exposure case. Total non-dietary mean values were 0.002 µg/kgbw/day (0.000; 0.004 µg/kgbw/day for 5th and 95th percentile, respectively) for BPA and 0.597 µg/kgbw/day (0.116 µg/kgbw/day and 1.506 µg/kgbw/day for 5th and 95th percentile, respectively) for DEHP. Indoor environments and especially dust ingestion were the main non-dietary contributors to the total exposure of BPA and DEHP with 60% and 81%. However, as expected, diet showed the higher contribution to total exposure with > 99.9% for BPA and 63% for DEHP. Although diet was considered the primary source of exposure to BPA and phthalates, it must be taken into account that with non-dietary sources the first-pass metabolism is lacking, so these may be of equal or even higher toxicological relevance than dietary sources. The present study is in the framework of "Health and environmental-wide associations based on large population surveys" (HEALS) project (FP7-603946).

Prenatal exposure estimation of BPA and DEHP using integrated external and internal dosimetry: A case study.

Prenatal exposure to Endocrine disruptors (EDs), such as Bisphenol A (BPA) and di (2-ethylhexyl) phthalate (DEHP), has been associated with obesity and diabetes diseases in childhood, as well as reproductive, behavioral and neurodevelopment problems. The aim of this study was to estimate the prenatal exposure to BPA and DEHP through food consumption for pregnant women living in Tarragona County (Spain). Probabilistic calculations of prenatal exposure were estimated by integrated external and internal dosimetry modelling, physiologically based pharmacokinetic (PBPK) model, using a Monte-Carlo simulation. Physical characteristic data from the cohort, along with food intake information from the questionnaires (concentrations of BPA and DEHP in different food categories and the range of the different food ratios), were used to estimate the value of the total dietary intake for the Tarragona pregnancy cohort. The major contributors to the total dietary intake of BPA were canned fruits and vegetables, followed by canned meat and meat products. In turn, milk and dairy products, followed by ready to eat food (including canned dinners), were the most important contributors to the total dietary intake of DEHP. Despite the dietary variations among the participants, the intakes of both chemicals were considerably lower than their respective current tolerable daily intake (TDI) values established by the European Food Safety Authority (EFSA). Internal dosimetry estimates suggest that the plasma concentrations of free BPA and the most important DEHP metabolite, mono (2-ethylhexyl) phthalate (MEHP), in pregnant women were characterized by transient peaks (associated with meals) and short half-lives (< 2h). In contrast, fetal exposure was characterized by a low and sustained basal BPA and MEHP concentration due to a lack of metabolic activity in the fetus. Therefore, EDs may have a greater effect on developing organs in young children or in the unborn child.

Bisphenol A analogues (BPS and BPF) present a greater obesogenic capacity in 3T3-L1 cell line.

This study was aimed at comparing the toxicity effects on cell viability and the obesogenic activity of Bisphenol A (BPA) and its analogues, Bisphenol S (BPS) and Bisphenol F (BPF), by in vitro assays with a preadipocytic 3T3-L1 cell line. To compare the toxic potential and select the concentrations of each chemical not showing a decrease in cell viability, MTT assay was performed. The cell phenotype was determined in differentiated 3T3-L1 adipocytes by red oil O staining. To determine the expression levels of the different adipogenic proteins the Western Blot test was performed. The results from MTT assay showed a greater toxic effect of BPA - at equal and even lower concentrations-than its analogues. However, BPS followed by BPF showed a greater neutral lipid storage capacity than BPA, which was reflected in the increase of the protein expression of CCAAT/enhancer binding protein α (C/EBPα), peroxisome proliferator-activated receptor gamma γ (PPARγ) and acid-binding protein 4 (FABP4). In summary, these BPA analogues -especially BPS- present a greater endocrine potential activity than BPA.

Experimental contamination of a closed endotracheal suction system: 24 h vs 72 h.

OBJECTIVE: To ascertain the desirability of replacing closed suction systems after 72 h rather than after 24 h (manufacturer's recommendations) because it is possible that a reduction in the frequency of manipulations might reduce the risk of exogenous nosocomial pneumonia. We investigated the presence of time-dependent differences (after 24 h and 72 h) in pathogen survival/growth in artificially contaminated closed suction catheters (OptiFlo). DESIGN: The trial simulated bacterial contamination of the airways using a suspension of 2 x 10(3) CFU/ml of Staphylococcus aureus or Pseudomonas aeruginosa. Contamination was performed on a total of 80 catheters. Forty were contaminated a total of eight times every 45-60 min. Another 40 catheters underwent the same procedure 24 times over three consecutive days. Microbiological analysis of the catheters took place after 24 h and 72 h, respectively. RESULTS: The mean S. aureus load was 9.4 CFU/catheter after eight suction procedures and 6.2 CFU/catheter after 24 suction procedures (3 days). Mean growth of P. aeruginosa was 5.3 CFU/catheter, and 8.2 CFU/catheter after 3 days. There was no statistically significant difference between day 1 and 3 for S. aureus (p = 0.474), but there was for P. aeruginosa (p = 0.004). CONCLUSION: Our findings show that, from an experimental point of view, it remains controversial whether routine change of closed suction catheters can be extended from 24 h to 72 h. However, clinical evidence suggests that prolonged use of a closed suctioning system is safe.

[Haemophilia and knee function: are there differences between haemophilic and healthy children?]. / Hämophilie und Kniefunktion - Gibt es Unterschiede zwischen hämophilen und gesunden Kinder?

With early prophylactic treatment our haemophilic children grow up in good health. Nevertheless, we cannot prevent every bleeding. Those bleedings may be just subclinical but they could lead to overloading of the knee and more and more of the ankle joint in the long term. Motion analysis can help to understand this process and prevent it. A comparison of the gait function of haemophilic and healthy children of the age 3-18 years showed distinct functional differences especially in the youngest age group (3-6 years). Apparently, the coordination skill gait rhythm was significantly worse in the heamophilic group. All measured functional deficits can be treated with physiotherapy. Possible reasons for these early functional differences are overprotection and/or early subclinical bleedings.

Control of cell growth by c-Myc in the absence of cell division.

The c-Myc protein (Myc) is a transcription factor, and deregulated expression of the c-myc gene (myc) is frequently found in tumours. In Burkitt's lymphoma (BL), myc is transcriptionally activated by chromosomal translocation. We have used a B-cell line called P493-6 that carries a conditional myc allele to elucidate the role of Myc in the proliferation of BL cells. Regulation of proliferation involves the coordination of cell growth (accumulation of cell mass) and cell division [1] [2] [3]. Here, we show that division of P493-6 cells was strictly dependent on the expression of the conditional myc allele and the presence of foetal calf serum (FCS). More importantly, cell growth was regulated by Myc without FCS: Myc alone induced an increase in cell size and positively regulated protein synthesis. An increase in protein synthesis is thought to be one of the causes of cell mass increase. Furthermore, Myc stimulated metabolic activities, as indicated by the acidification of culture medium and the activation of mitochondrial enzymes. Our results confirm the model that Myc is involved in the regulation of cell growth [4] and provide, for the first time, direct evidence that Myc induces cell growth, that is, an increase in cell size, uncoupled from cell division.

Concentrations of polychlorinated biphenyls (PCBs) and polybrominated diphenyl ethers (PBDEs) in milk of women from Catalonia, Spain.

In this study, the concentrations of polychlorinated biphenyls (PCBs) and polybrominated diphenyl ethers (PBDEs) in milk from women living in the vicinity of a new hazardous waste incinerator (HWI) in Catalonia, Spain, were determined. The study was performed after 4 years of regular operations in the facility and the present PCB levels were compared with baseline concentrations obtained in a pre-operational program. PCBs and PBDEs levels were determined by HRGC/HRMS in 15 samples. In the present study planar PCBs ranged from 1.3 to 6.3 pg WHO-TEQ/g fat with a mean value of 3.8 pg WHO-TEQ/g fat. After adding dioxin-like mono-ortho-PCBs the total PCB-TEQ concentrations ranged from 3.8 to 13.3 pg WHO-TEQ/g fat (mean value: 8.7 pg WHO-TEQ/g fat). A comparison of the current data with those obtained in the baseline study showed significant decreases for both planar and total WHO-TEQ of PCBs: 47.9% and 44.6%, respectively. PCB concentrations in milk of women living in urban zones were higher than those living near industrial areas (10.1 and 7.4 pg WHO-TEQ/g fat, respectively). Mean PBDE concentrations were 2.2 and 2.5 ng/g fat for women living in urban and industrial zones, respectively. Dietary intake of PCBs and PBDEs for a standard adult woman samples were 898 and 843 ng/day for PCBs, and 72 and 63 ng/day for PBDEs, for residents in urban and industrials areas, respectively. This study suggests that dietary intake is more relevant for human exposure to PCBs and PBDEs than living near the HWI.

The transcriptional program of a human B cell line in response to Myc.

The proto-oncogene c-myc (myc) encodes a transcription factor (Myc) that promotes growth, proliferation and apoptosis. Myc has been suggested to induce these effects by induction/repression of downstream genes. Here we report the identification of potential Myc target genes in a human B cell line that grows and proliferates depending on conditional myc expression. Oligonucleotide microarrays were applied to identify downstream genes of Myc at the level of cytoplasmic mRNA. In addition, we identified potential Myc target genes in nuclear run-on experiments by changes in their transcription rate. The identified genes belong to gene classes whose products are involved in amino acid/protein synthesis, lipid metabolism, protein turnover/folding, nucleotide/DNA synthesis, transport, nucleolus function/RNA binding, transcription and splicing, oxidative stress and signal transduction. The identified targets support our current view that myc acts as a master gene for growth control and increases transcription of a large variety of genes.

Molecular characterization of metal-binding polypeptide domains by electrospray ionization mass spectrometry and metal chelate affinity chromatography.

Metal ion-binding of synthetic peptides containing HxH and CxxC motifs was investigated by electrospray ionization mass spectrometry (ESI-MS) and metal chelate affinity chromatography. A high affinity of Ni2+ and Cu2+ to HxH containing sequences was found. Based on their natural metal ion-binding potential it was possible to include metal affinity chromatography in the purification process of two proteins without using an additional His-tag sequence: ATPase-439, a P type ATPase from Helicobacter pylori and the amyloid precursor protein (APP).

Biomass production and nitrogen content of C3- and C4- grasses in pure and mixed culture with different nitrogen supply.

Two C3 grasses (Hordeum vulgare L., Avena sativa L.) and two C4 grasses (Panicum miliaceum L., Panicum crus-galli L.) were cultivated in standard soil in the open air in pure cultures and in various mixed cultures at low and high nitrogen fertilization levels. After three months the dry weight, length and nitrogen content of the aboveground and below-ground parts of the plants and the shoot/root ratios were determined. Hordeum vulgare was the most successful species irrespective of the nitrogen fertilization level, and also exhibited in most cases the highest nitrogen concentrations. Panicum miliaceum, on the other hand, was the species least able to compete. The production of biomass was reduced in cultures growing under nitrogen starvation conditions, this phenomenon being more pronounced with respect to the C4 than to the C3 species. The decrease in the production of biomass at low N conditions was most drastic with Panicum crus-galli, the species with the lowest nitrogen content and thus assumed to be best adapted to nitrogen starvation conditions. In cultures growing at low nitrogen fertilization levels the shoot/root ratios of all species.shifted in favour of an increasing root proportion. The extent of this shift, however, differed from species to species.

PCDD/F levels in the neighbourhood of a municipal solid waste incinerator after introduction of technical improvements in the facility.

In 1998 and 1999, the concentrations of polychlorinated dibenzo-p-dioxins and dibenzofurans (PCDD/Fs) were determined in soil and herbage samples collected in the vicinity of an old municipal solid waste incinerator (MSWI) (S. Adrià del Besòs, Barcelona, Spain). Just after the 1999 collection, an adaptation to the EU legislation on pollutant emissions from the stack was carried out in this facility. The purpose of the present study was to determine the current concentrations of PCDD/Fs in soil and herbage samples collected in the neighbourhood of the MSWI and to compare these concentrations with those obtained in the 1998 and 1999 surveys. During the period 1998-1999, an increase of 31% (P>.05) was found in the median PCDD/F levels in soils, while a reduction of 40% (P>.05) was observed in the period 1999-2000. Similarly, in the period 1998-1999 an increase of 41% (P>.05) was found in the levels of PCDD/Fs in vegetation, while a 30% decrease (P<.05) was seen in the period 1999-2000. Although after introduction of the technical improvements in the MSWI a notable reduction in the levels of PCDD/Fs in soil and vegetation has been noted, the median decreases have not been as great as it could be expected according to the very pronounced reductions in PCDD/F emissions from the stack. It indicates that other emission sources of PCDD/Fs also have a notable impact on the area under direct influence of the MSWI.

Annual variation in the levels of metals and PCDD/PCDFs in soil and herbage samples collected near a cement plant.

In May 2000, the levels of a number of metals (As, Cd, Co, Cr, Cu, Hg, Mn, Ni, Pb, Sn, Tl, V and Zn) and polychlorinated dibenzo-p-dioxins (PCDDs) and polychlorinated dibenzofurans (PCDFs) were determined in soil and herbage samples collected near a cement plant from Sta. Margarida i els Monjos (Catalonia, Spain). To determine the temporal variation in the concentrations of metals and PCDD/PCDFs, in May 2001 soil and herbage samples were again collected at the same sampling points and analyzed for the levels of metals and PCDD/PCDFs. In general terms, metal concentrations in soils did not change between May 2000 and May 2001, while significant decreases in the levels of Cr, Ni and V were found in herbage. On the other hand, no significant differences in the mean I-TEQ values of PCDD/PCDFs were found in soil and herbage samples. The results of this survey show that according to the annual variation in the levels of metals and PCDD/PCDFs the environmental impact of the cement plant on the area under its direct influence is not relevant.

Spatial distribution and temporal variation of metals in the vicinity of a municipal solid waste incinerator after a modernization of the flue gas cleaning systems of the facility.

In June 1994 and 1997, the concentrations of a number of elements were determined in soil and herbage samples collected in the vicinity of a municipal solid waste incinerator (MSWI) (Tarragona, Catalonia, Spain). In August 1997, an adaptation to the EU legislation on pollutant emissions from the stack was carried out to the incinerator. In June 1999, soil and herbage samples were collected again at the same sampling points and the levels of arsenic (As), cadmium (Cd), chromium (Cr), lead (Pb), manganese (Mn), mercury (Hg), nickel (Ni) and vanadium (V) were measured by ICP-MS or AAS with graphite furnace. The results are compared with those obtained in the 1994 and 1997 surveys. In the period 1997-1999, the only significant changes in soil levels corresponded to Cd and Pb, with decreases of 21.0% and 53.5%, respectively. In vegetation, only Mn levels showed a significant reduction, which contrasts with the notable increases found in the concentrations of As, Hg and Ni. According to the results of this survey, it seems evident that other metal emission sources in the same area of study are masking the environmental improvements carried out in the MSWI.

Zinc and copper levels in serum and urine: relationship to biological, habitual and environmental factors.

Zinc and copper levels were determined in serum and urine of 434 subjects living in an industrial and an agricultural area of Tarragona Province, Spain. Zinc and copper concentrations were related to a range of factors such as sex, age, blood pressure, and drinking and smoking habits. Geometric mean serum zinc and copper concentrations were, respectively, 113.9 and 84 micrograms dl-1, while the mean values for urine zinc and copper concentrations were 698.7 and 26.6 micrograms g-1 creatinine. Serum zinc and copper levels and urine copper concentrations in men were significantly lower than in women, while there were no differences in serum or urinary zinc and copper levels with age. The consumption of alcohol significantly reduced the levels of zinc and copper in serum, whereas blood pressure had no influence on these values. The levels of zinc and copper in urine were not affected by the smoking and drinking habits, place of residence, or blood pressure. In general terms, the results of this study agree with previously reported values from different countries.

Levels of PAHs in soil and vegetation samples from Tarragona County, Spain.

The levels of 16 polycyclic aromatic hydrocarbons (PAHs) were determined in 24 soil and 12 wild chard samples collected in Tarragona County (Catalonia, Spain), an area with an important number of chemical and petrochemical industries. Samples were also collected in urban/residential zones and in presumably unpolluted sites (control samples). In soils, the sum of the 16 PAHs ranged between 1002 and 112 ng/g (dry weight) for samples collected near chemical industries and unpolluted sites, respectively. With the exception of acenaphthylene, acenaphthene, anthracene and benzo[k]fluoranthene, no significant differences in the levels of the remaining PAHs were found among the different zones of sample collection. In chard samples, the highest value (sum of 16 PAHs) was observed in the residential area, followed by the industrial and the unpolluted zones, with concentrations of 179, 58 and 28 ng/g (dry weight), respectively. In general terms, the current PAH concentrations in soil and vegetation are lower than the levels reported in a number of investigations from different regions and countries. They are also below the maximum PAH concentrations allowed by the Catalan legislation for different uses of soil.

Metal pollution of soils and vegetation in an area with petrochemical industry.

The aim of this study was to determine the concentrations of arsenic (As), cadmium (Cd), chromium (Cr), mercury (Hg), manganese (Mn), lead (Pb), and vanadium (V) in soil and chard samples collected in various industrial sites of Tarragona County (Spain), an area with an important number of petrochemical industries. Samples were also collected in urban (Tarragona downtown) and presumably unpolluted (blank samples) sites. Human health risks derived from metal inhalation and ingestion of soils were also assessed. With the exception of an increase in the levels of Cd and V, significant differences in soil samples from the industrial and the unpolluted zone were not found. In chard samples, significant differences between collection sites were only noted for V concentrations. For non-carcinogenic risks, the current levels of metals in the industrial area were lower than those considered as safe for the general population. In relation to carcinogenic risks, only As ingestion and Cr inhalation in the industrial zone might potentially cause an increase of the cases of cancer. A Kohonen self-organized map (an Artificial Neural Network) showed differences in metal concentrations according to the zone of origin of the samples. The current results suggest that although in general terms the petrochemical complex is not a relevant metal pollution source for the area, attention should be paid to As, Cr and V.

Lead in children's hair, as related to exposure in Tarragona Province, Spain.

Concentrations of lead were determined in the hair of 478 school children living in an industrial and an agricultural area of Tarragona Province, Spain. The influence of age, sex, hair colour and family occupation on the children's hair lead concentration was also evaluated. Girls had more lead in their hair than boys (10.54 vs 6.55 micrograms g-1), and the lead levels decreased with age. Lead concentrations in hair with respect to hair colour decreased in the order red, blond, brown, black. Family occupation was one of the most significant factors influencing the lead content of children's hair. Although the average lead content was higher in the industrial area (9.38 micrograms g-1) than in the agricultural area (7.80 micrograms g-1), this difference was not statistically significant.

Mercury in hair for a child population from Tarragona Province, Spain.

Mercury concentrations were determined in scalp hair of 233 school children aged 6-16 years. The study was carried out in three communities (Flix, Tarragona and Tortosa) from Tarragona Province (Southern Catalonia, Spain). The influence of the variables place of residence, age, sex, fish and seafood consumption, number of dental amalgam fillings, hair color, parents' occupation, and smoking habits of the household members was also examined. The geometric mean mercury concentration in hair was 0.77 microgram/g. The place of residence, sex, and the frequency in consuming fish and seafood were the variables that significantly affected hair mercury concentrations. Girls had more mercury in their hair than boys, whereas hair mercury levels were significantly correlated with the frequency in the fish and seafood consumption, with the levels being more elevated when the fish and seafood consumption was also higher. Hair mercury concentrations were also affected by the place of residence, with school children of Flix showing lower mercury concentrations than those found in children from Tarragona and Tortosa. The remaining variables had no influence on hair mercury levels.

Dietary intake of copper, chromium and zinc in Tarragona Province, Spain.

The dietary intake of copper, chromium, and zinc from foods in Tarragona Province (NE Spain) was determined by means of two different methods: the total diet study and the duplicate diet study. In the total diet study, a total of 375 food samples were analyzed, whereas duplicates of food consumed weekly by 20 families were analyzed for copper, chromium and zinc concentrations. Pulses were the group with the highest levels of the three elements; vegetables and fruits which are basic items in the 'Mediterranean diet', contained low concentrations of these metals. Total intakes of copper, chromium and zinc were 1156.3, 124.6 and 7522.9 micrograms day-1, respectively. On the other hand, the total intake of the elements obtained through the duplicate diet study was 1119.8 micrograms day-1 for copper, 129.0 micrograms day-1 for chromium, and 6800.1 micrograms day-1 for zinc. The daily intake of copper and zinc by the population of Tarragona Province is lower than the usually recommended values, while the intake of chromium is closer to those values.