Gene/Protein Disease Symptom Drug Enzyme Compound
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Query: EC:2.7.10.2 (focal adhesion kinase)
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1,2-Diaminobenzene and its derivatives react with selenous acid in acidic solution to form the piazselenols, which can be extracted into toluene. Microgram amounts of selenium can be determined spectrophotometrically by measuring the absorbance of these piazselenols extracted into toluene. A more sensitive method, in which the piazselenols extracted into toluene are detected by electron-capture gas chromatography, has been developed. In order to find a more sensitive reagent, 13 piazselenols were synthesized. The retention behaviour and sensitivity in electron-capture detection gas chromatography and the distribution ratios between aqueous solution and toluene were studied for each piazselenol extracted into toluene. Of these piazselenols, 4,6-dibromopiazselenol, formed by the reaction of 1,2-diamino-3,5-dibromobenzene with selenous acid, was found to be best as regards sensitivity and distribution ratio. Under the optimal conditions for the formation and the extraction of the piazselenol, the practical detection limit was 1 ng. Selenium(VI) and total selenium in NBS Bovine Liver, SRM 1577, were determined successfully.
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PMID:Some 1,2-diaminobenzene derivatives as reagents for gas chromatographic determination of selenium with an electron-capture detector. 88 62

An isotope dilution gas chromatography-mass spectrometry method for selenium determination in urine using 76Se as an internal standard is described. Three different derivatizing reagents, 4-nitro-o-phenylenediamine (NPD), 3,5-dibromo-o-phenylenediamine (DBPD), and 4-trifluoromethyl-o-phenylenediamine (TFMPD) were investigated for their gas chromatographic behavior including memory and precision and accuracy of isotope ratio measurements. By these criteria, the performance of these reagents was TFMPD greater than DBPD greater than NPD. Overall precision values of 1 to 7% were observed in determining Se isotope ratios at the 10-ng level, with no significant difference in using any of the three reagents. Memory effect was observed in the order NPD greater than DBPD greater than TFMPD with TFMPD showing no measurable memory effect. Accuracy of the GC-MS method was verified by quantitation of selenium in the NIST freeze-dried urine reference material SRM 2670.
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PMID:Determination of selenium in urine by isotope dilution gas chromatography-mass spectrometry using 4-nitro-o-phenylenediamine, 3,5-dibromo-o-phenylenediamine, and 4-trifluoromethyl-o-phenylenediamine as derivatizing reagents. 151 65

We have improved Zeeman atomic absorption spectrometric determination of selenium in serum by using an acidic solution of Ag + Cu + Mg as a matrix modifier and performing the charring step in flowing O2. Under these conditions, we could calibrate serum results with selenium standards in bovine albumin solutions. Mean analytical recovery was 98%, and the CV was 1.8% within runs, 2.9% between runs. Analysis of Reference Materials from the U.S. National Bureau of Standards (SRM 909 and RM 8419) yielded the values of 106 (SD 2.4) and 15 (SD 1.6) microgram/L, respectively, in good agreement with the expected values (106 and 16, respectively). The method--being reliable and relatively simple and rapid--is suitable for use in epidemiological screenings. Mean selenium concentrations in serum sampled from 274 adults in central Italy were 90 (SD 15) microgram/L. For 11-year-old children, these values were lower and showed a tendency to sex-related difference: 82 (SD 9.9) microgram/L for 97 boys, 78 (SD 9.3) microgram/L for 90 girls.
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PMID:Improved determination of selenium in serum by Zeeman atomic absorption spectrometry. 333 41

A microbiological assay to detect different chemical compounds of selenium for potential future use in the study of the distribution of these chemical forms in foods is being developed. This assay is based on the detection, by infrared analysis, of CO2 in a culture of Escherichia coli when the bacteria are grown in the presence of various selenium compounds. The CO2 production is the result of selenium-dependent formate dehydrogenase activity, which catalyzes oxidation of formic acid produced during glucose metabolism. Smooth response curves were generated over several orders of magnitude for selenocystine, selenite, and selenomethionine. The assay detects selenium concentrations (above background) as low as 1.5 nM for selenocystine and selenite and 4 nM for selenomethionine in minimal medium. Detection of selenomethionine was enhanced (to a sensitivity of 1.5 nM) by the addition of methionine to minimal medium and was enhanced even further (to a sensitivity of 0.8 nM) by the addition of a defined mixture of amino acids. Selenomethionine could be assayed in the presence of an amino acid concentration which is proportional to the amino acid/elemental selenium ratio found in a wheat gluten reference material (NIST SRM 8418). This implies that the assay can detect selenium compounds in a variety of foods at low concentrations, avoiding the background CO2 production caused by high concentrations of non-selenium-containing amino acids. The observation that methionine enhanced selenomethionine availability for formate dehydrogenase synthesis supports studies in animals demonstrating that methionine controls selenomethionine incorporation into selenoenzymes.(ABSTRACT TRUNCATED AT 250 WORDS)
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PMID:Optimization of an Escherichia coli formate dehydrogenase assay for selenium compounds. 781 Oct 71

Electrothermal atomic absorption spectrometry with a temperature-stabilizing platform and palladium(II) nitrate-magnesium nitrate-barium nitrate matrix modifier combination was used to measure total selenium concentrations in 0.5-ml samples of human urine. The method was validated by analysis of desiccated, standard urine samples (SRM 2670) from NIST. The peak height response was linear from 0 to 150 micrograms Se/liter urine with a sensitivity of 0.001 absorbance unit/micrograms Se/liter and a detection limit of 6 micrograms Se/liter (P = 0.05) for a single measurement. Total urinary selenium concentrations and urine volumes of 24-h samples from 28 healthy adult subjects on self-selected diets and living in San Angelo, Texas were measured. Daily total urinary excretions of 16 male and 12 female subjects (mean +/- SD) were 28 +/- 26 and 34 +/- 25 micrograms Se/day, respectively. A single subject taking 300 micrograms Se/day in the form of aqueous SeO2 as a dietary supplement excreted 232 micrograms Se/day. The unsupplemented subjects' daily selenium excretions correspond to a mean dietary intake of approximately 60 micrograms/day.
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PMID:Palladium(II), magnesium(II), and barium(II) nitrate combinations for matrix modification in electrothermal atomic absorption measurement of total selenium in human urine. 797 75

An on-line anodic stripping voltammetry (ASV) flow system, interfaced with inductively coupled plasma atomic emission spectrometry (ICP-AES) and inductively coupled plasma mass spectrometry (ICPMS) detectors, has been used for determination of arsenic(III) and selenium(IV) and for elimination of polyatomic interferences which arise from chloride in sample matrices. Details of the working electrode preparation are discussed. Arsenic signals in ICP-AES were enhanced by as much as 10 times through preconcentration of sample volumes up to 5 mL. Using ICP-AES detection, recoveries for analyte spikes in 1:10 diluted urine were 102% for As(III) (matrix-matched standards) and 91% for Se(IV) (standards in electrolyte). Using ICPMS detection, determination of certified Se(IV) and Se(IV) spikes in diluted NIST SRM 2670 elevated urine gave recoveries of 92-103%, while recoveries of As(III) spikes in diluted NIST SRM 2670 urine ranged from 94 to 113%. High levels of chloride matrix exhibited little effect on the arsenic signal with ICP-AES or ICPMS detection. Elimination of the polyatomic interference ArCl+ in ICPMS was very efficient for diluted NIST SRM 2670 urine and for a synthetic matrix of 1000 micrograms/mL chloride.
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PMID:Determination of arsenic(III) and selenium(IV) using an on-line anodic stripping voltammetry flow cell with detection by inductively coupled plasma atomic emission spectrometry and inductively coupled plasma mass spectrometry. 829 28

Structural modifications to the photoinactive benzophenoxazine Nile blue A have led to three novel derivatives which include 5-ethylamino-9-diethylaminobenzo[a]phenoxazinium (EtNBA), 5-ethylamino-9-diethylaminobenzo[a]phenothiazinium (EtNBS), and 5-ethylamino-9-diethylaminobenzo[a]phenoselenazinium (EtNBSe) chlorides. The incorporation of sulfur and selenium into the benzophenoxazine moiety results in lipophilic, red-absorbing (650-660 nm) chromophores which possess significantly increased singlet oxygen yields (0.025 and 0.65, respectively, compared to 0.005 for EtNBA). This study examines the photosensitizing efficacies and pharmacokinetics in vitro in the EMT-6 murine mammary sarcoma cell line as well as the physicochemical, photochemical, and redox properties of these new analogues. Comparisons with Photofrin II, the only photosensitizer available clinically, were made in an attempt to high-light their different pharmacological characteristics. The photodynamic activity of the benzophenoxazine dyes correlates with their ability to generate the phototoxin singlet oxygen and increases in the following order: EtNBA < EtNBS << EtNBSe. At an extracellular dye concentration of 0.5 microM, the light dose required to kill approximately 50% of the cells was 2.0 and < 0.5 J/cm2 for the sulfur and selenium dyes, respectively. The light dose required to kill approximately 50% of the cells for both EtNBA and Photofrin II could not be determined because of their weak phototoxic effect under these conditions. At a light dose of 3.3 J/cm2, EtNBSe is approximately 1000 times more phototoxic than Photofrin II. All three benzophenoxazine derivatives are characterized by a similar uptake/efflux pattern in vitro consisting of a rapid and extensive cellular accumulation followed by a slow efflux rate. Contrary to their rapid uptake, 50% of the accumulated EtNBS and EtNBSe is retained intracellularly after a 6-h period in dye-free medium. Video-enhanced fluorescence microscopy corroborates the rapid uptake measurements as well as indicating the intracellular localization of the dyes in both living and thermally inactivated cells. Low extracellular dye concentrations (0.05 microM) result in a punctate fluorescence pattern in the perinuclear region, while higher dye concentrations (> 0.1 microM) lead to additional fluorescence in the cytoplasm, cytomembranes, and other organelles but apparently not the nucleus. Absorption spectrometry revealed that living cells rapidly reduce the dyes to their colorless leuko form (photoinactive) if oxygen is not readily available in the environment. It is shown that the cellular reduction is an enzymatic process and that an oxygen-free and cell-free medium containing both the coenzyme NADH and the hydride transfer enzyme diaphorase is capable of reducing the dyes to the colorless leuko form.(ABSTRACT TRUNCATED AT 400 WORDS)
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PMID:Phototoxicity, redox behavior, and pharmacokinetics of benzophenoxazine analogues in EMT-6 murine sarcoma cells. 849 21

Hydride generation atomic absorption spectrometry was used to determine selenium content in cereals, legumes and dry fruits from the coast of the province of Granada (southeastern Spain). Accuracy was assured using both a NIST SRM 1572 and recovery experiments. The precision expressed as relative standard deviation (R.S.D.) varied between 6.50% for seeds and 15.98% for bread. The highest selenium concentrations were found for dry fruits (294.6 ng/g), followed by legumes (111.8 ng/g), and the lowest for cereals (27.8 ng/g). Considering the average daily individual consumption of these foods in Andalusia (southern Spain), the daily dietary intake of selenium supplied by this source is 15.36 micrograms/day for an adult. The content of total selenium in corn samples taken from the zone is independent of both human and industrial activities (P > 0.05).
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PMID:Determination of selenium in cereals, legumes and dry fruits from southeastern Spain for calculation of daily dietary intake. 864 54

The method described was developed to be applied in determination of selenium in biological matrices (plasma, urine and tissues) using ETA-AAS with Zeeman background correction. These matrices were obtained from non-fasting S.D. rats and Beagle dogs of both sexes in order to acquire data on the endogenous levels of selenium in these laboratory animals when fed with standard diets. For tissue digestion, a simple procedure using the strong organic base, Soluene 350, was adopted. Precision assays were carried out monitoring Se(IV) levels in spiked matrices (range from 25 to 200 ng) and obtaining relative standard deviations (RSD%) in the range from 3.2% to 14.5% (intra-day) and from 7.6% to 15.9% (inter-day). Accuracy assays gave relative errors (RE%) in the range from -6.5 to 4.2% (intra-day) and from -5.5% to 5.7% (inter-day). The validity of the method was checked on reference material (NBS SRM 1577 bovine liver) and the values obtained correlated with the certified ones. The detection limit assumed was 0.9 ng/ml, whereas the quantitation limit of selenium in matrices ranged from 2 to 5 ng/ml (or g), depending on the kind of sample.
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PMID:Determination of selenium in plasma, urine and tissues, of standard diet-fed rats and dogs by ETA-atomic absorption spectroscopy with Zeeman background correction. 903 61

In 1996, the National Institute of Standards and Technology (NIST) released Standard Reference Material 1846 (Infant Formula), which can be used as a control material for assigning values to in-house control materials and for validating analytical methods for measurement of proximates, vitamins, and minerals in infant formula and similar matrixes. The SRM was manufactured by preparing a spray-dried formula base containing fat, protein, carbohydrates, and minerals and then combining that formula base with a dry-blend vitamin premix that supplied the vitamins. The Certificate of Analysis for SRM 1846 provides assigned values for concentrations of proximates (fat, protein, etc.), vitamins, and minerals for which product labeling is required by the Infant Formula Act of 1980 and by the Nutrition Labeling and Education Act of 1990. These assigned values were based on agreement of measurements by NIST and/or collaborating laboratories. Certified values are provided for vitamins A (trans), E, C, B2, and B6 and niacin. Noncertified values are provided for solids, ash, fat, nitrogen, protein, carbohydrate, calories, vitamin D, delta-tocopherol, gamma-tocopherol, vitamin B1, vitamin B12, folic acid, pantothenic acid, biotin, choline, inositol, calcium, phosphorus, magnesium, iron, zinc, copper, sodium, potassium, and chloride. Information values are provided for iodine, manganese, selenium, and vitamin K.
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PMID:Certification of nutrients in Standard Reference Material 1846: infant formula. 917 Jun 57


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