Determination of homovanillic, isohomovanillic and vanillylmandelic acids in human urine by means of glass capillary gas—liquid chromatography with temperature-programmed electron-capture detection

The determination of urinary homovanillic, isohomovanillic and vanillylmandelic acids as their trifluoroacetylhexafluoroisopropyl ester derivatives by glass capillary gas—liquid chromatography has been studied. It was shown that even with high column efficiencies a single peak—single compound relationship could not be assumed and for reliable quantitation it was necessary to check determinations with a second gas—liquid chromatography column.     

The contribution of neighbouring countries to pesticide levels in Dutch surface waters

Compared with other European countries, Dutch consumption of pesticides is high, particularly in agriculture, with many of the compounds found in surface waters in high concentrations and various standards being exceeded. Surface water quality is routinely monitored and the data obtained are published in the Dutch Pesticides Atlas. One important mechanism for reducing pesticide levels in surface waters is authorisation policy, which proceeds on the assumption that the pollution concerned has taken place in the Netherlands. The country straddles the delta of several major European rivers, however, and as river basins do not respect national borders some of the water quality problems will derive from neighbouring countries. Against this background the general question addressed in this article is the following: To what extent do countries neighbouring on the Netherlands contribute to pesticide pollution of Dutch surface waters? To answer this question, data from the Pesticides Atlas for the period 2005-2009 were used. Border zones with Belgium and Germany were defined and the data for these zones compared with Dutch data. In the analyses, due allowance was also made for authorised and non-authorised compounds and for differences between flowing and stagnant waters. Monitoring efforts in the border zones and in the Netherlands were also characterised, showing that efforts in the former are similar to those in the rest of the country. In the border zone with Belgium the relative number of non-authorised pesticides exceeding the standards is clearly higher than in the rest of the Netherlands. These exceedances are observed mainly in flowing waters. In contrast, there is no difference in the relative number of standard-exceeding measurements between the border zones and the rest of the Netherlands. In the boundary zones the array of standard-exceeding compounds clearly deviates from that in the rest of the Netherlands, with compounds authorised in the neighbouring countries but not in the Netherlands, such as flufenacet, featuring prominently. The share of the neighbouring countries in the total number of exceedances in the Netherlands is roughly proportional to the relative area of the border zones. Although there is a certain influx of pesticides from across national borders, the magnitude of the problem appears to be limited.     

Similarities and differences between measured and predicted concentrations of pesticides in Dutch surface waters

In order to have a thorough evaluation of the progress and effectiveness of Dutch crop protection policy, both model predictions and measured pesticide concentrations in surface waters are considered. To this purpose, monitoring data obtained by various water boards and other monitoring institutes were processed. Data were aggregated over a two year time period and over space (at 1x1 km-grid). A geographic view is given in the Dutch Pesticides Atlas (www.pesticidesatlas.nl). The model used for the predictions was the Dutch National Environmental Indicator NMI version 2 (www.nmi.alterra.nl) that has input data regarding spray drift data, crop interception, soil and climate and many more. Information on aggregation steps over time and space, grid sizes, information on crop areas was geared to one another for both instruments. Results on measured pesticide concentrations in surface waters and model predictions were compared to each other at the national scale. For this study, 10 different cases were selected covering a large range of pesticides' characteristics and pesticides' use. In 60% of the cases, the results were largely in agreement with each other when expressed as absolute numbers of measurements exceeding the environmental quality standard. This is very accurate and useful for policy purposes. Based on concentrations and on the order of magnitude, no significant agreement between measurements and model predictions was found. Differences were explained by various factors, and an overview of predominant systematic differences between the measurements and the model predictions was presented. Using both measurements and model predictions in supporting environmental policy evaluations is warranted, because of higher Weight-of-Evidence. Combining both can assist in optimizing the knowledge on pesticides behaviour, fate and ecological problems and therefore this is the preferred evaluation method.     

Aluminium in human breast tissue

Aluminium is omnipresent in everyday life and increased exposure is resulting in a burgeoning body burden of this non-essential metal. Personal care products are potential contributors to the body burden of aluminium and recent evidence has linked breast cancer with aluminium-based antiperspirants. We have used graphite furnace atomic absorption spectrometry (GFAAS) to measure the aluminium content in breast biopsies obtained following mastectomies. The aluminium content of breast tissue and breast tissue fat were in the range 4-437 nmol/g dry wt. and 3-192 nmol/g oil, respectively. The aluminium content of breast tissue in the outer regions (axilla and lateral) was significantly higher (P=0.033) than the inner regions (middle and medial) of the breast. Whether differences in the regional distribution of aluminium in the breast are related to the known higher incidence of tumours in the outer upper quadrant of the breast remains to be ascertained.     

Increased autocrine production of insulin-like growth factor II (IGF-II) alters serum sensitivity of MCF-7 human breast cancer cell proliferation

Abstract. Regulation of the growth of breast cancer cells is the result of a complex interaction between steroid hormones and growth factors, and in particular of oestrogen and insulin-like growth factors (IGF). Alteration of any one mitogenic component can affect the cell response to other pathways. Previous work has shown that increased autocrine production of IGF-II from a transfected inducible expression vector can result in reduced oestrogen sensitivity of growth of MCF-7 human breast cancer cells. This report describes alterations to non-oestrogen regulated pathways of cell growth following enhanced IGF-II expression in these transfected MI7 cells. Serum sensitivity of cell growth in the absence of oestrogen was found to differ between MI7 and untransfected MCF-7 cells, in that growth of MI7 but not MCF-7 cells was strongly inhibited by high serum levels. Increased serum had no effect on levels of IGF-II mRNA, IGFIR, IGFBP4 mRNA, or IGFBP secreted in MI7 cells. However, growth inhibition by serum in MI7 cells could be overcome by increasing levels of IGF-II in the serum or by removal of IGFBP onto polycarbonate membranes. Thus, the growth inhibition by serum in MI7 cells is concluded to result from the increased levels of IGFBP added with higher serum. This would support an inhibitory role for IGFBP on growth of breast cancer cells when cell growth is being driven by IGF pathways in the absence of oestrogen, and would suggest that cellular sensitivity to such factors can depend on levels of endogenous IGF production.     

Regional and Elevational Patterns in Vascular Epiphyte Richness on an East Asian Island

The distribution of species on mountains has been related to various predictor variables, especially temperature. Thermal specialization—presumed to be more pronounced on tropical mountains than on temperate mountains—accounts for the elevational pattern of species richness and varies between organisms and geographic areas. In this study, the elevational and regional distribution patterns of 331 epiphyte species in Taiwan were explored using 39,084 botanic collections, mostly from herbaria. Species richness showed a peak in elevation at 500–1500 m. This peak could not be explained by a null model, the mid‐domain effect, suggesting that environmental variables accounted mostly for the distribution of species on the mountains. Next, species distributions were modeled to assess epiphyte regional and elevational distribution patterns. The model results not only corroborated the position of the mid‐elevation peak in richness, but also identified two mountain areas on the island with exceptionally high species richness. These areas of high epiphyte diversity coincide with areas of high rainfall in relation to the direction of the prevailing winds. Moreover, a subsequent exploratory ordination analysis showed a varied thermal preference between epiphyte subcategories (hemiepiphytes, dicotyledons, orchids, and ferns). In contrast to predictions by the elevational Rapoport's rule, ordination analysis also showed that the degree of thermal specialization increased with elevation, suggesting that highland species may be especially vulnerable to global warming.     

Substrate Specificity of Cytoplasmic N-Glycosyltransferase

N-Linked protein glycosylation is a very common post-translational modification that can be found in all kingdoms of life. The classical, highly conserved pathway entails the assembly of a lipid-linked oligosaccharide and its transfer to an asparagine residue in the sequon NX(S/T) of a secreted protein by the integral membrane protein oligosaccharyltransferase. A few species in the class of γ-proteobacteria encode a cytoplasmic N-glycosylation system mediated by a soluble N-glycosyltransferase (NGT). This enzyme uses nucleotide-activated sugars to modify asparagine residues with single monosaccharides. As these enzymes are not related to oligosaccharyltransferase, NGTs constitute a novel class of N-glycosylation catalyzing enzymes. To characterize the NGT-catalyzed reaction, we developed a sensitive and quantitative in vitro assay based on HPLC separation and quantification of fluorescently labeled substrate peptides. With this assay we were able to directly quantify glycopeptide formation by Actinobacillus pleuropneumoniae NGT and determine its substrate specificities: NGT turns over a number of different sugar donor substrates and allows for activation by both UDP and GDP. Quantitative analysis of peptide substrate turnover demonstrated a strikingly similar specificity as the classical, oligosaccharyltransferase-catalyzed N-glycosylation, with NX(S/T) sequons being the optimal NGT substrates.     

3,5,3′,5′-Tetrachlorobiphenyl is a weak oestrogen agonist in vitro and in vivo

Polychlorinated biphenyls (PCBs) are widespread, persistent environmental contaminants of which some congeners can act as endocrine disrupters. Previous work has shown that 3,4,3′,4′-tetrachlorobiphenyl (PCB77) can act as an oestrogen with actions mediated through the oestrogen receptor. Here, oestrogenic actions have been assessed for two further tetrachlorobiphenyl isomers. Assays of oestrogenic action have involved (1) ligand regulation of oestrogen-sensitive gene expression; (2) ligand regulation of cell growth in oestrogen-dependent human breast cancer cell lines MCF7 McGrath and ZR-75-1; and (3) ligand activity in the immature mouse uterine weight bioassay in vivo. These results demonstrate that 3,5,3′,5′-tetrachlorobiphenyl (PCB 80) can be considered to be a weak oestrogen agonist, but the 2,5,2′,5′-congener (PCB 52) revealed no oestrogenic properties in any of these assays. Implications of these results are discussed in relation to structure-activity predictions for environmental oestogens.     

Multihormone regulation of MMTV-LTR in transfected T-47-D human breast cancer cells

Multihormonal regulation on the long terminal repeat (LTR) region of mouse mammary tumour virus (MMTV) has been studied using T-47-D human breast cancer cells stably transfected with the steroid sensitive LTR-C3 chimaeric gene. The specificity of steroid action on transfected LTR sequences has been compared with regulation of endogenous cellular markers. We conclude that the hormone response element of the LTR can be induced by physiological concentrations of androgen, progestin and glucocorticoid. 17 beta-Oestradiol did not regulate the LTR at physiological levels but an effect was found at 10(-6) M. This effect was not inhibited by antioestrogen nor was it reproduced by the synthetic oestrogen diethylstilboestrol suggesting such effects do not occur via the oestrogen receptor. The antioestrogens tamoxifen and transhydroxytamoxifen do not induce the LTR. No significant steroid competition was found in LTR regulation: whilst oestradiol did not act at physiological concentration it did not interfere with induction by androgen, progestin or glucocorticoid. Such gene regulation did not simply follow receptor status of the cells nor was it reflected in patterns of growth regulation by steroids. The implications of these findings on the mechanism of steroid hormone action are discussed.