Solubilization of cholesterol and polycyclic aromatic compounds into sodium bile salt micelles (part 2).
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The aqueous solubility of cholesterol was determined over the temperature range from 288.2 to 318.2 K with intervals of 5 K by the enzymatic method. The solubility was (3.7+/-0.3)x10(-8) mol dm(-3) (average +/- S.D.) at 308.2 K. The maximum additive concentrations of cholesterol into the aqueous micellar solutions of sodium deoxycholate (NaDC), sodium ursodeoxycholate (NaUDC), and sodium cholate (NaC) were spectrophotometrically determined at different temperatures. The cholesterol solubility increased in the order of NaUDC+info)
Relationship of exposure to coke-oven emissions and urinary metabolites of benzo(a)pyrene and pyrene in coke-oven workers.
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Coke-oven workers are occupationally exposed to a high concentration of polycyclic aromatic hydrocarbons (PAH). r-7,t-8,9,c-10-Tetrahydroxy-7,8,9,10-tetrahydrobenzo(a)pyrene (trans-anti-BaP-tetraol) and 1-hydroxypyrene (1-OHP) are urinary metabolites of benzo(a)pyrene and pyrene, respectively. In this study, we investigated the relationship among individual air exposure to benzene soluble fraction (BSF) of total particulates, as a surrogate marker of ambient PAH exposures, and urinary trans-anti-BaP-tetraol and 1-OHP concentrations in coke-oven workers at a steel plant in Taiwan. Fifty-seven subjects, including 41 male workers who work in one coke-oven plant and 16 men (referents) from an administrative area, were studied. The mean trans-anti-BaP-tetraol and 1-OHP concentrations (mean +/- SD) were 0.4 +/- 0.3 nmol/mol creatinine and 9.7 +/- 21.6 micromol/mol creatinine, respectively, in coke-oven workers. These levels were significantly higher than those in referents (0.03 +/- 0.03 nmol/mole creatinine, P < 0.001 and 0.4 +/- 0.2 micromol/mol creatinine, P < 0.01, respectively). Urinary trans-anti-BaP-tetraol concentrations were significantly and positively correlated with individual average BSF and urinary 1-OHP concentrations. That is, the higher the urinary trans-anti-BaP-tetraol concentrations, the more ambient BSF exposure and urinary 1-OHP concentrations (Spearman correlation coefficients r = 0.68 and 0.70, respectively; P < 0.0001; n = 57). These findings suggest that urinary 1-OHP and trans-anti-BaP-tetraol might be considered as potential biomarkers for the assessment of uptake of known PAH carcinogens in the air. (+info)
The atherogen 3-methylcholanthrene induces multiple DNA adducts in mouse aortic smooth muscle cells: role of cytochrome P4501B1.
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OBJECTIVE: 3-Methylcholanthrene (MC), a polycylic aromatic hydrocarbon, induces atherogenesis in mice fed an atherogenic diet. In this study, we tested the hypothesis that MC would induce DNA adducts in mouse aortic smooth muscle cells (SMCs) and that cytochrome P4501B1 (CYP1B1) plays an important role in the activation of MC to genotoxic intermediates. METHODS: Cultured SMCs were treated with MC or the vehicle dimethyl sulfoxide (DMSO), and DNA was isolated after 24 h. In some experiments, the cells were pre-treated with the CYP1B1 inhibitor 1-ethynylpyrene (EP) prior to exposure to MC. DNA adducts were determined by the 32P-postlabeling assay. Aryl hydrocarbon hydroxylase assay was measured by fluorimetry. RESULTS: MC induced formation of 12 DNA adducts that were not observed in DMSO-treated cells. DNA adduct formation was dose-dependent, with maximum response observed at 3 microM. Pre-treatment of cells with EP dramatically suppressed DNA adduct formation by MC. MC treatment caused induction of CYP1B1, but not CYP1A1. CONCLUSION: The induction of high levels of multiple DNA adducts in SMCs by MC suggests that SMCs have a functional enzymatic machinery capable of metabolically activating MC to genotoxic metabolites. The significant inhibition by EP of MC-induced DNA adduct formation indicated that CYP1B1 was the primary CYP enzyme responsible for formation of genotoxic metabolites that may play a role in the induction of atherosclerosis by MC. (+info)
Fluorescent molecular sensory system based on bis pyrene-modified gamma-cyclodextrin dimer for steroids and endocrine disruptors.
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A gamma-cyclodextrin dimer modified with two pyrene moieties, 6-(2-pyrenebutylate-aminoethyl)pyrenebutylate-amino-6-deoxy-bis-gamma-cyclodextri n, has been synthesized in the presence of N,N'-dicyclohexycarbodiimide from gamma-cyciodextrin dimer linked with ethylenediamine at an upper rim. The sensing ability and binding property of the titled dimer were investigated for bile acids and endocrine disruptors. This cyclodextrin dimer showed both monomer and excimer fluorescence; the guest-induced emissions were observed as increases or decreases depending on the guest. The values deltaI(m)/I0(m) and deltaI(ex)/I0(ex), where I0(m) and I(m) are fluorescence intensities of monomer emission in the absence and presence of a guest and I0(ex) and I(ex) are those of excimer emission and deltaI(m) and deltaI(ex) were I(m) - I0(m) and I(ex) - I0(ex), respectively, were used as a parameter of sensitivity. This host exhibited highly sensitive molecular recognition ability for bile acids and endocrine disruptors, in which the sensing parameters obtained from monomer emission were plus or minus values, whereas the parameters obtained as excimer emission were minus ones. The behavior of the appended moieties of the host during a host-guest complexation was studied by induced circular dichroism (ICD) and fluorescence spectra. The ICD intensities of the titled dimer were decreased upon an addition of a guest. The guest-induced variations in the fluorescence and ICD intensity suggest that the appended moieties move by altering the spatial relationship in the hydrophobic space between two linked cyclodextrins. (+info)
Liquid chromatographic determination of ornithine and lysine based on intramolecular excimer-forming fluorescence derivatization.
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A highly sensitive and selective fluorometric determination method for ornithine and lysine has been developed. This method is based on an intramolecular excimer-forming fluorescence derivatization with a pyrene reagent, 4-(1-pyrene)butyric acid N-hydroxysuccinimide ester (PSE), followed by reversed-phase liquid chromatography (LC). The analytes, containing two amino moieties in a molecule, were converted to the corresponding dipyrene-labeled derivatives by reaction with PSE. The derivatives afforded intramolecular excimer fluorescence (450-550 nm) which can clearly be discriminated from the normal fluorescence (370-420 nm) emitted from PSE and monopyrene-labeled derivatives of monoamines. The structures of the derivatives and the emission of excimer fluorescence were confirmed by LC with mass spectrometry and with three-dimensional fluorescence detection system, respectively. The PSE derivatives of ornithine and lysine could be separated by reversed-phase LC on ODS column with isocratic elution. The detection limits (signal-to-noise ratio = 3) for ornithine and lysine were 3.5 and 3.7 fmol, respectively, for a 20-microl injection. Furthermore, this method had enough selectivity and sensitivity for the determination of ornithine and lysine in normal human urine. (+info)
Derivative matrix isopotential synchronous fluorescence spectroscopy for the direct determination of 1-hydroxypyrene as a urinary biomarker of exposure to polycyclic aromatic hydrocarbons.
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Urinary 1-hydroxypyrene is a biomarker in the measurement of human exposure to polycyclic aromatic hydrocarbons. A rapid and simple derivative isopotential synchronous fluorescence method was developed for the direct determination of 1-hydroxypyrene in urine. A length of iso-intensity route was scanned on the three-dimensional fluorescence spectrum of urine and this result was combined with that from derivative technique. Thus the strong background signals of urine were removed and the 1-hydroxypyrene can be determined directly in urine without tedious pre-separation. The derivative isopotential synchronous fluorescence spectrum was directly obtained from a single scan on a spectrofluorometer, which further simplified isopotential synchronous fluorescence technique. The recoveries of 93% to 115% were obtained for 1-hydroxypyrene added to urine. (+info)
Ca-dependent binding of actin to gelsolin.
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Ca(2+) of 0.3-1.0 microM induces both the exposure of tryptic cleavage sites within the gelsolin molecule inaccessible in the Ca-free conformation, and binding of one actin monomer to the N-terminal half of gelsolin. On the other hand, gelsolin-induced enhancement of pyrene actin fluorescence was observed only above 50 microM Ca(2+), and a ternary actin/gelsolin complex preformed in 200 microM Ca(2+) was stable only above 30 microM Ca(2+). These results provide direct evidence for Ca-induced transitions from closed to open conformation of the gelsolin molecule in the range of 3 x 10(-7) to 10(-6) M Ca(2+). They also suggest that Ca(2+)>10(-5) M is required to stabilize actin-actin contacts in the 2:1 actin/gelsolin complex. (+info)
Urinary 1-hydroxypyrene as a biomarker of internal dose of polycyclic aromatic hydrocarbons in carbon black workers.
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In this study, a total of 30 workers were selected, including eight wet pelletizing workers and 22 packaging workers. For all selected workers, urine samples were collected on the first day pre-shift, first day post-shift and fifth day post-shift, and their urinary 1-hydroxylpyrene levels (1-OHP) were determined (denoted as BM1pre, BM1post and BM5post, respectively). Personal respiratory exposures, including both inhalable particle-bound PAHs (Cinh) and gaseous PAHs (Cgas), together with dermal exposure to particle-bound PAHs (Cskin) were measured. Personal background information, including age, sex and smoking habit, was carefully registered. Pyrene exposure was statistically significantly correlated with exposure to PAHs and carcinogenic PAHs. Multiple linear regression analysis results showed that the BM1post values could not be explained by workers' exposures. For BM5post in packaging workers, both the regression model (R2 = 0.73) and the regression coefficients for Cgas, Cinh and Cskin were statistically significant (P < 0.05). For pelletizing workers, the R2 value was higher but was not statistically significant because of the smaller number of workers. The resultant regression coefficients for 'sex', 'smoking habit' and 'age' were statistically insignificant (P >> 0.05), which could be because these variables made relatively small contributions to BM5post. In conclusion, this study suggests BM5post could be a suitable indicator for PAH exposures of carbon black workers, on the condition that both respiratory (including gaseous PAHs and particle-bound PAHs) and dermal exposures have been assessed. (+info)