Gene/Protein Disease Symptom Drug Enzyme Compound
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Query: EC:3.1.6.1 (sulfatase)
 3,205 document(s) hit in 31,850,051 MEDLINE articles (0.00 seconds)

Metabolites of 3'-methyl-4-(dimethylamino)azobenzene (3'-Me-DAB) in the rat bile and urine were investigated by the use of a tracer technique. 3H-3'-Me-DAB in cottonseed oil was administered orally by a stomach tube. The dye metabolites in the bile and urine collected during 24 hr after the administration were hydrolyzed with beta-glucuronidase/arylsulfatase. The hydrolyzed metabolites were then extracted with chloroform or separated by chromatography on Amberlite XAD-2 using methanol as a solvent. The metabolites in the chloroform or methanol eluates were identified by the reverse isotope dilution analysis, before or after separation by thin-layer chromatography. The N-demethylated, aryl hydroxylated, and their azo-reduced products were detected in the bile, in addition to the products oxidized at the ring methyl group as the new metabolites. On the other hand, the metabolites retaining the azo-linkage were scarcely detected in urine and instead 3-aminobenzoic acid, 3-amino-6-hydroxytoluene, and their N-acetylated products were major metabolites in urine. These results indicate that the metabolism of 3'-Me-DAB in the rat involves oxidation of the ring methyl group. Significance of the ring methyl group in the carcinogenic action of aminoazo dyes is also discussed.
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PMID:Analysis of biliary and urinary metabolites of 3'-methyl-4-(dimethylamino)azobenzene in rats. 10 70

[14C] Cholesterol-5 alpha, 6 alpha-expoxide, administered to mice by either gastric intubation or skin painting, was rapidly and primarily excreted in the feces. Residual amonts of the epoxide and its metabolites were found in a wide variety of organs, and persisted for at least 72 hr. At some sites (principally the liver, the small intestinal contents and the combined stomach/duodenum and their contents), the labeled compound existed in a water-soluble form which could not be extracted with chloroform/methanol. Treatment of the small intestinal contents with a preparation of beta-glucuronidase/sulfatase produced a marked increase in the amount of organic-solvent-extractable cholesterol-alpha-epoxide and other polar metabolites. Unchanged epoxide was found mainly in the feces and the skin at the site of application. On the basis of these results, stool specimens, and not blood samples, should be analyzed to detect the presence of this compound and/or its metabolites in vivo.
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PMID:The metabolic fate of cholesterol-5 alpha, 6 alpha-expoxide in vivo. 48 Nov 35

A yellow-colored protein (YCP) was isolated from the hemolymph (i.e. blood) of fifth instar wandering stage larvae of Manduca sexta. The molecular mass of YCP was 31 kDa by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. Gel filtration chromatography suggested that native YCP was a monomer. The absorbance spectrum of YCP revealed maxima at 278 and 405 nm. Chromophore was released from YCP through denaturation of the protein with methanol and chloroform. In neutral solution and in acid, the released chromophore showed the absorbance characteristics of an ommochrome: ommatin D. In addition, the chromophore was sensitive to treatment with arylsulfatase as would be expected for ommatin D. The amino acid composition and the N-terminal sequence of YCP were determined. The YCP polypeptide chain was found to be glycosylated. Carbohydrate analysis suggested that Man and GlcNAc were present in a 3:1 ratio. Circular dichroism indicated that YCP consisted of 68% beta-pleated sheet with no alpha-helices being detected. An in vitro incubation of larval fat body in the presence of [35S]methionine indicated that this organ was the site of synthesis. Ommochromes arise in insects as end products of the metabolism of tryptophan. It is well-documented that ommochromes occur in both the tissues and the excreta of insects. We propose that in M. sexta, one such tryptophan metabolite is found in the hemolymph associated with a specific protein.
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PMID:Purification and properties of an ommochrome-binding protein from the hemolymph of the tobacco hornworm, Manduca sexta. 193 73

A reversed-phase high-performance liquid chromatographic method is described, which allows the simultaneous quantification of propranolol and 4-hydroxypropranolol enantiomers in human plasma. After extraction from plasma (pH 10.5) using ethyl acetate, the enantiomers are derivatized with R-(+)-phenylethylisocyanate as chiral derivatization reagent and triethylamine as basic catalyst in chloroform. Ascorbic acid is used to prevent 4-hydroxypropranolol from oxidation during the extraction. Chromatographic separation on ODS columns and fluorescence detection (228 nm/greater than 340 nm) allows sensitive quantitation of all derivatives. Incubation of the plasma samples with beta-glucuronidase/arylsulfatase and the use of the specific beta-glucuronidase inhibitor saccharo-1,4-lactone allows the quantitation of both the sulfate and glucuronide conjugates of the enantiomers. The method was applied to human plasma samples from a subject after administration of 60 mg racemic propranolol three times daily.
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PMID:Simultaneous determination of propranolol and 4-hydroxypropranolol enantiomers after chiral derivatization using reversed-phase high-performance liquid chromatography. 238 82

A previously unknown melatonin metabolite was isolated by chloroform extraction and reverse phase HPLC from human and rat urine after administration of synthetic melatonin and characterized by mass spectroscopy and proton magnetic resonance spectroscopy to 1-acetyl-1,2,3,3a,8,8a-hexahydro-8a-hydroxy-5-methoxypyrrolo[2,3-b ]indole, a cyclic isomer of 2-hydroxymelatonin. This isolation was based on the fact that our melatonin antibody (a-MT-K1) cross-reacted against this novel metabolite at a level of 0.1% (melatonin 100%). In our HPLC program for indoles the cyclic 2-hydroxymelatonin eluted at 25 min, separately from synthetic indoles, between 6-hydroxymelatonin (19 min) and melatonin (35 min). In [3H] melatonin studies it was found to be present (at 25 min in our HPLC), accounting for 5% of the urinary metabolites of melatonin in the rat. Since beta-glucuronidase-arylsulfatase treatment of rat urine did not liberate the cyclic 2-hydroxymelatonin this would appear to be excreted into urine as the free form.
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PMID:A cyclic isomer of 2-hydroxymelatonin: a novel metabolite of melatonin. 356 38

Peripheral blood leukocytes isolated from men and women were studied for their capacity to metabolize estrone (E1) sulfate. Fresh human leukocytes (granulocytes and mononuclear cells) were incubated in phosphate buffer, pH 7.4, containing [3H]E1S for 1 h at 37 C. The samples were extracted with chloroform for measurement of the [3H]E1 formed, and the results were corrected for nonenzymatic hydrolysis. The mean E1 sulfatase activity in leukocytes isolated from normal women in the follicular phase of their cycle was 75% higher than that during the luteal [1840 +/- 179 (+/- SE) vs. 1048 +/- 101 fmol E1 micrograms protein-1 h-1; P less than 0.004] and higher than that in normal men (875 +/- 123; P less than 0.002), but was not different from that in menopausal (1349 +/- 151) or hirsute women (1700 +/- 222). In pregnant women, the mean leukocyte E1 sulfatase activity was significantly lower (861 +/- 147) than that in nonpregnant women in the follicular phase (P less than 0.003). These results suggest that progesterone may modulate E1 sulfatase activity, whereas estrogens do not.
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PMID:Variations in estrone sulfatase activity in human leukocytes. 366 72

A sensitive, specific, and reliable liquid chromatography tandem mass spectrometry (LC-MS/MS) method was developed for detection and identification of zeranol in chicken or rabbit liver. A homogenized liver sample was hydrolyzed with beta-glucuronidase/arylsulfatase, and the hydrolysate was extracted with ethyl ether. The supernatant was evaporated to dryness, and the residue was dissolved in chloroform and re-extracted with sodium hydroxide. After acidification, the extract was cleaned up on a C18 solid-phase extraction cartridge and analyzed by electrospray LC-MS/MS in the negative ion mode. The multiple reaction monitoring transition from both m/z 321 to 277 and m/z 321 to 303 was monitored for confirmation, and the product ion of 277 was used for quantitation. Separation was performed on a Waters XTettra C18 column (50 x 2.1 mm, 3.5 microm) combined with a safeguard column (Symmetry C18, 20 x 3.9 mm, 5 microm), using a gradient elution with acetonitrile and 20 mM ammonium acetate. Calibration curves were prepared and good linearity was achieved over the concentration ranges tested. For all liver samples fortified at 3 different levels of 1, 5, and 50 microg/kg, the overall recoveries and relative standard deviations were in the range of 61-90 and 8-13%, respectively. The limit of quantitation based on the assay validation was 1 microg/kg. The method had been used on a routine basis for detection and identification of zeranol in liver samples.
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PMID:Detection and identification of zeranol in chicken or rabbit liver by liquid chromatography-electrospray tandem mass spectrometry. 1218 Jun 76

A sensitive analytical method was developed for quantitative analysis of delta(9)-tetrahydrocannabinol (delta(9)-THC), 11-nor-delta(9)-tetrahydrocannabinol-carboxylic acid (delta(9)-THC-COOH), cannabinol (CBN) and cannabidiol (CBD) in human hair. The identification of delta(9)-THC-COOH in hair would document Cannabis use more effectively than the detection of parent drug (delta(9)-THC) which might have come from environmental exposure. Ketamine was added to hair samples as internal standard for CBN and CBD. Ketoprofen was added to hair samples as internal standard for the other compounds. Samples were hydrolyzed with beta-glucuronidase/arylsulfatase for 2h at 40 degrees C. After cooling, samples were extracted with a liquid-liquid extraction procedure (with chloroform/isopropyl alcohol, after alkalinization, and n-hexane/ethyl acetate, after acidification), which was developed in our laboratory. The extracts were analysed before and after derivatization with pentafluoropropionic anhydride (PFPA) and pentafluoropropanol (PFPOH) using a Hewlett Packard gas chromatographer/mass spectrometer detector, in electron impact mode (GC/MS-EI). Derivatized delta(9)-THC-COOH was also analysed using a Hewlett Packard gas chromatographer/mass spectrometer detector, in negative ion chemical ionization mode (GC/MS-NCI) using methane as the reagent gas. Responses were linear ranging from 0.10 to 5.00 ng/mg hair for delta(9)-THC and CBN, 0.10-10.00 ng/mg hair for CBD, 0.01-5.00 ng/mg for delta(9)-THC-COOH (r(2)>0.99). The intra-assay precisions ranged from <0.01 to 12.40%. Extraction recoveries ranged from 80.9 to 104.0% for delta(9)-THC, 85.9-100.0% for delta(9)-THC-COOH, 76.7-95.8% for CBN and 71.0-94.0% for CBD. The analytical method was applied to 87 human hair samples, obtained from individuals who testified in court of having committed drug related crimes. Quantification of delta(9)-THC-COOH using GC/MS-NCI was found to be more convenient than GC/MS-EI. The latter may give rise to false negatives due to the detection limit.
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PMID:Hair analysis for delta(9)-THC, delta(9)-THC-COOH, CBN and CBD, by GC/MS-EI. Comparison with GC/MS-NCI for delta(9)-THC-COOH. 1220 25

BACKGROUND: The purpose of this study was to localize the expression of steroid sulfatase (STS) in cumulus cells and to determine the relationship between STS mRNA expression and the serum levels of follicle-stimulating hormone (FSH), luteinizing hormone (LH), estradiol and progesterone. METHODS: The subject group included 49 women (29 to 44 years old) for whom in vitro fertilization treatment was indicated. All subjects gave informed consent. One hundred fourteen samples of cumulus-oocyte complex (COC) were obtained under microscopic observation. Part of the COC was stained by STS antibody. RNA was extracted by phenol-chloroform method and real-time PCR was performed. Serum of each patient was collected and was measured by ELISA. RESULTS: Some of the cumulus samples were stained by STS antibody. The expression of STS mRNA in all samples was confirmed by quantitative RT-PCR. Although there was no significant correlation between the level of STS mRNA and the serum levels of estradiol, progesterone and LH, there was a statistically significant negative correlation between the level of STS mRNA expression and the serum level of FSH (n = 105, p = 0.018, r = -0.22). CONCLUSION: These results have demonstrated for the first time the expression of STS in cumulus cells by immunohistological stainings and real-time RT-PCR. STS expression in cumulus cells may be related to the control of the local steroidal environment in the oocyte. Serum FSH may control STS mRNA expression from the results of RT-PCR, although the correlation was low.
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PMID:Localization and gene expression of steroid sulfatase by RT-PCR in cumulus cells and relationship to serum FSH levels observed during in vitro fertilization. 1582 1

A mass spectrometric method is described for monitoring cerebrosides in the presence of excess concentrations of alkali metal salts. This method has been adapted for use in the assay of arylsulfatase A (ASA) and the cerebroside sulfate activator protein (CSAct or saposin B). Detection of the neutral glycosphingolipid cerebroside product was achieved via enhancement of ionization efficiency in the presence of lithium ions. Assay samples were extracted into the chloroform phase as for the existing assays, dried, and diluted in methanol-chloroform-containing lithium chloride. Samples were analyzed by electrospray ionization mass spectrometry with a triple quadrupole mass spectrometer in the multiple reaction monitoring tandem mass spectrometric mode. The assay has been used to demonstrate several previously unknown or ambiguous aspects of the coupled ASA/CSAct reaction, including an absolute in vitro preference for CSAct over the other saposins (A, C, and D) and a preference for the non-hydroxylated species of the sulfatide substrate over the corresponding hydroxylated species. The modified assay for the coupled ASA/CSAct reaction could find applicability in settings in which the assay could not be performed previously because of the need for radiolabeled substrate, which is now not required.
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PMID:A novel mass spectrometric assay for the cerebroside sulfate activator protein (saposin B) and arylsulfatase A. 1606 47


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