Long-term Data from the Swedish National Environmental Monitoring Program of Pesticides in Surface Waters.

Environmental monitoring is essential for assessing the impact of human activities on the environment. Monitoring data are used to ascertain that environmental standards are met, to inform policy making, to determine trends, and to provide parameterization data for prediction models. The design of monitoring programs depends on what is being monitored, for what purpose, and available resources. Here we describe the strategy and design of the Swedish environmental monitoring program for chemical pesticides in surface waters and provide data generated within this program since 2002 (). We include examples of how the data can be used for toxicity assessments, trend analyses, and comparison between sampling strategies. Our goal is to increase awareness of this dataset and provide detailed information about the data so that it may be incorporated into meta-analytical research, comparison studies, model validation, and other scientific efforts.

Sequence analysis of heparan sulfate epitopes with graded affinities for fibroblast growth factors 1 and 2.

Proteins that belong to the fibroblast growth factor (FGF) family regulate proliferation, migration, and differentiation of many cell types. Several FGFs, including the prototype factors FGF-1 and FGF-2, depend on interactions with heparan sulfate (HS) proteoglycans for activity. We have assessed tissue-derived HS fragments for binding to FGF-1 and FGF-2 to identify the authentic saccharide motifs required for interactions. Sequence information on a range of N-sulfated HS octasaccharides spanning from low to high affinity for FGF-1 was obtained. All octasaccharides with high affinity for FGF-1 (> or =0.5 m NaCl required for elution) contained an internal IdoUA(2-OSO(3))-GlcNSO(3)(6-OSO(3))-IdoUA(2-OSO(3))-trisaccharide motif. Octasaccharides with a higher overall degree of sulfation but lacking the specific trisaccharide motif showed lower affinity for FGF-1. FGF-2 was shown to bind to a mono-O-sulfated HS 6-mer carrying a single internal IdoUA(2-OSO(3))-unit. However, a di-O-sulfated -IdoUA(2-OSO(3))-GlcNSO(3)-IdoUA(2-OSO(3))-trisaccharide sequence within a HS 8-mer gave stronger binding. These findings show that not only the number but also the positions of individual sulfate groups determine affinity of HS for FGFs. Our findings support the notion that FGF-dependent processes can be modulated in vivo by regulated expression of distinct HS sequences.

Binding of heparin/heparan sulfate to fibroblast growth factor receptor 4.

Fibroblast growth factors (FGFs) are heparin-binding polypeptides that affect the growth, differentiation, and migration of many cell types. FGFs signal by binding and activating cell surface FGF receptors (FGFRs) with intracellular tyrosine kinase domains. The signaling involves ligand-induced receptor dimerization and autophosphorylation, followed by downstream transfer of the signal. The sulfated glycosaminoglycans heparin and heparan sulfate bind both FGFs and FGFRs and enhance FGF signaling by mediating complex formation between the growth factor and receptor components. Whereas the heparin/heparan sulfate structures involved in FGF binding have been studied in some detail, little information has been available on saccharide structures mediating binding to FGFRs. We have performed structural characterization of heparin/heparan sulfate oligosaccharides with affinity toward FGFR4. The binding of heparin oligosaccharides to FGFR4 increased with increasing fragment length, the minimal binding domains being contained within eight monosaccharide units. The FGFR4-binding saccharide domains contained both 2-O-sulfated iduronic acid and 6-O-sulfated N-sulfoglucosamine residues, as shown by experiments with selectively desulfated heparin, compositional disaccharide analysis, and a novel exoenzyme-based sequence analysis of heparan sulfate oligosaccharides. Structurally distinct heparan sulfate octasaccharides differed in binding to FGFR4. Sequence analysis suggested that the affinity of the interaction depended on the number of 6-O-sulfate groups but not on their precise location.

Selectively desulfated heparin inhibits fibroblast growth factor-induced mitogenicity and angiogenesis.

Fibroblast growth factors (FGFs) are known to induce formation of new blood vessels, angiogenesis. We show that FGF-induced angiogenesis can be modulated using selectively desulfated heparin. Chinese hamster ovary cells (CHO677) deficient in heparan sulfate biosynthesis were employed to assess the function of heparin/heparan sulfate in FGF receptor-1 (FGFR-1) signal transduction and biological responses. In the presence of FGF-2, FGFR-1 kinase and subsequent mitogen-activated protein kinase Erk2 activities were augmented in a dose-dependent manner, whereas high concentrations of heparin resulted in decreased activity. The length of the heparin oligomer, minimally an 8/10-mer, was critical for the ability to enhance FGFR-1 kinase activity. The N- and 2-O-sulfate groups of heparin were essential for binding to FGF-2, whereas stimulation of FGFR-1 and Erk2 kinases by FGF-2 also required the presence of 6-O-sulfate groups. Sulfation at 2-O- and 6-O-positions was moreover a prerequisite for binding of heparin to a lysine-rich peptide corresponding to amino acids 160-177 in the extracellular domain of FGFR-1. Selectively 6-O-desulfated heparin, which binds to FGF-2 but fails to bind the receptor, decreased FGF-2-induced proliferation of CHO677 cells, presumably by displacing intact heparin. Furthermore, FGF-2-induced angiogenesis in chick embryos was inhibited by 6-O-desulfated heparin. Thus, formation of a ternary complex of FGF-2, heparin, and FGFR-1 appears critical for the activation of FGFR-1 kinase and downstream signal transduction. Preventing complex formation by modified heparin preparations may allow regulation of FGF-2 functions, such as induction of angiogenesis.

Structural basis and potential role of heparin/heparan sulfate binding to the angiogenesis inhibitor endostatin.

Recombinant mouse endostatin produced by mammalian cells was shown to bind to heparin with a K(d) of 0.3 microM, suggesting that this interaction may play a role in its anti-angiogenic activity. Alanine mutagenesis demonstrated that a major site of four clustered arginines (positions 155, 158, 184 and 270) and a second site (R193, R194) are essential for binding. The same epitopes also participate in endostatin binding to heparan sulfate and sulfatides but not in its binding to the extracellular protein ligands fibulin-1 and fibulin-2. Analyses with various heparin fragments demonstrated a minimum size (12mer) for efficient binding to endostatin and a crucial role of 2-O- and 6-O-sulfation. Furthermore, a substantial proportion (10-50%) of heparan sulfate chains obtained from various tissues showed a distinct binding to endostatin, indicating its potential to interact with extracellular and/or membrane-bound proteoglycans. Angiogenesis induced by basic fibroblast growth factor-2 (FGF-2), but not by vascular endothelial growth factor (VEGF), in a chick chorioallantoic membrane assay could be inhibited by endostatin in a dose-dependent manner. The mutational block of heparin binding decreased endostatin inhibition to low levels but elimination of zinc binding had no effect.

Characterization of fibroblast growth factor 1 binding heparan sulfate domain.

Fibroblast growth factors FGF-1 and FGF-2 mediate their biological effects via heparan sulfate-dependent interactions with cell surface FGF receptors. While the specific heparan sulfate domain binding to FGF-2 has been elucidated in some detail, limited information has been available concerning heparan sulfate structures involved in the recognition of FGF-1. In the current study we present evidence that the minimal FGF-1 binding heparan sulfate sequence comprises 5-7 monosaccharide units and contains a critical trisulfated IdoA(2-OSO3)-GlcNSO3(6-OSO3) disaccharide unit. N-Sulfated heparan sulfate decasaccharides depleted of FGF-1 binding domains showed dose-dependent and saturable binding to FGF-2. These data indicate that the FGF-1 binding domain is distinct from the minimal FGF-2 binding site, previously shown to contain an IdoA(2-OSO3) residue but no 6-O-sulfate groups. We further show that the FGF-1 binding heparan sulfate domain is expressed in human aorta heparan sulfate in an age-related manner in contrast to the constitutively expressed FGF-2 binding domain. Reduction of heparan sulfate O-sulfation by chlorate treatment of cells selectively impedes binding to FGF-1. The present data implicate the 6-O-sulfation of IdoA(2-OSO3)-GlcNSO3 units in cellular heparan sulfate in the control of the biological activity of FGF-1.

Pesticides in stream water within an agricultural catchment in southern Sweden, 1990-1996.

Pesticide loss to stream water was studied in a small agricultural catchment in southern Sweden during the period 1990-1996. A total of 38 pesticides were detected in water samples, including 30 herbicides, four fungicides, three insecticides and one metabolite of one of the herbicides. Concentrations of pesticides in stream water were observed throughout the sampling periods. Peak concentrations occurred during the spraying seasons and following runoff events. Daily average concentrations sometimes varied by one order of magnitude from one day to another. Pesticides were also found in water samples as a result of incautious actions during handling and application procedures. Concentrations were lower at the outlet of the catchment area when the water had passed an open part of the stream, compared to concentrations detected in discharge from a culvert system upstream. This was largely a result of dilution from groundwater intrusion during low-flow periods. Sampling at different sites along the culvert demonstrated that the small village situated in the catchment did not contribute to pesticide findings in the culvert discharge. Wind drift had little influence on stream-water quality. Pesticide application for weed control in farmyards resulted in a substantial contribution to the pesticide load in stream water. Pesticide were persistent in the discharge throughout the winter and originated from both autumn and spring applications, as well as from farmyard application. Some autumn applied pesticides prevailed in stream flow during the following summer. Total amounts of pesticides lost in stream flow during May-September each year varied between 0.5 and 2.8 kg during the 7-year period, corresponding to approximately 0.1% of the applied amount. Losses of single pesticides were generally less than 0.3% of the applied amount during individual years. Pesticides from agricultural applications in the catchment constituted, on average, 82% of the total transported amount lost during May-September each year, of which 2% was from autumn application the previous year. There was an overall correlation between amounts used in the catchment and occurrence in the water samples. The total pesticide load in water decreased markedly during the course of the investigation, in accordance with decreased amounts applied during spring and early summer. The results indicate that concentrations of some pesticides entering head-water streams in agricultural areas are close to, and during certain time periods even above those levels demonstrated as having an impact on the aquatic flora and fauna.

Identification of O-sulphate substituents on D-glucuronic acid units in heparin-related glycosaminoglycans using novel synthetic disaccharide standards.

The two disaccharides, methyl 4-O-(2-O-sulpho-beta-D-glucopyranosyl-uronic acid)-2-deoxy-2-amino-alpha-D-glucopyranoside and methyl 4-O-(3-O-sulpho-beta-D-glucopyranosyluronic acid)-2-deoxy-2-amino-alpha-D-glucopyranoside, were prepared by de novo synthesis, and converted to the corresponding 2,5-anhydro-D-[1-3H]mannitol derivatives by deamination with nitrous acid followed by reduction with NaB3H4. The resultant labelled products were used as standards in the identification, by anion-exchange high-performance liquid chromatography (HPLC), of disaccharides generated by HNO2/NaB3H4 treatment of heparan sulphate isolated from human brain. The two standards, containing 2-O- and 3-O-sulphated glucuronic acid, respectively, were clearly separated by the HPLC procedure. Comparison with the deamination products derived from heparan sulphate showed that the mono-O-sulphated disaccharide species containing a sulphated glucuronic acid unit co-eluted with the 2-O-sulphated standard. The corresponding component isolated from other heparan sulphate preparations, or from heparin, also eluted at the same position. No disaccharide derived from heparin or heparan sulphate appeared at the elution position of the 3-O-sulphated standard. It is concluded that D-glucuronic acid units in heparin-related glycosaminoglycans may be sulphated at C2, whereas no evidence has been found for sulphation at C3. By contrast, analysis of mono-O-sulphated disaccharides derived from a chemically sulphated, bacterial capsular polysaccharide (generated by Escherichia coli K5) clearly demonstrated the occurrence of O-sulphate groups at C-3 of D-glucuronic acid units.

Spirited team. Interview by Jill L. Sherer.

J. Harry Kreuger put an old calling to a new use. Kreuger, an ordained minister and pastoral care director at Adventist Healthcare Mid-Atlantic, Rockville, MD, is a self-proclaimed visionary. In his words, "I see things that could be and ask 'why not?'" Instead of turning to divine inspiration alone for a better way to create wellness in the Adventist system's service community, Kreuger asked his colleagues in neighboring churches for help. He spoke with staff editor Jill L. Sherer about how they responded to his initiative--and on how the 5-month-old Damascus (MD) Community Wellness Center became a reality.

Primary malignant fibrous histiocytoma of the lung. Fine needle aspiration cytologic features.

A case of primary malignant fibrous histiocytoma of the lung occurring in a 71-year-old woman is presented. The preoperative aspiration cytology showed a large-cell, undifferentiated, malignant neoplasm suggestive of carcinoma. Subsequent histologic examination revealed a primary malignant fibrous histiocytoma. The diagnosis was confirmed by electron microscopic and immunohistochemical studies. Cytologic features of this rare primary pulmonary sarcoma are discussed.

Sialic acid in rabbit lacrimal gland fluid.

To date, there has been no attempt to determine which of the orbital glands contribute the sialic acid, which has been found in tears and tear mucoids. In the present study, sialic acid was found in the fluid collected directly from the lacrimal gland excretory duct, uncontaminated by the secretions of the other orbital glands (Nictitans, Harderian, conjunctival) as well as in the fluid secreted into the conjunctival sac by the other orbital glands, uncontaminated by lacrimal gland fluid. At all flow rates, the rate of secretion of sialic acid increased as flow rate increased in both fluids and the rate of secretion of sialic acid by the lacrimal gland was three times that by the other orbital glands. This is the first demonstration that a substance, which can be derived from either nonserum glycoproteins, such as the tear mucoids, or alpha-globulins, is a component of the secretions of the lacrimal gland, as well as of the secretion of the other orbital glands.