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The inducible isoform of nitric oxide synthase (iNOS) and three zinc tetrathiolate mutants (C104A, C109A, and C104A/C109A) were expressed in Escherichia coli and purified. The mutants were found by ICP-AES and the zinc-specific PAR colorimetric assay to be zinc free, whereas the wild-type iNOS zinc content was 0.38 +/- 0.01 mol of Zn/mol of iNOS dimer. The cysteine mutants (C104A and C109A) had an activity within error of wild-type iNOS (2.24 +/- 0.12 micromol of NO min(-1) mg(-1)), but the double cysteine mutant had a modestly decreased activity (1.75 +/- 0.14 micromol of NO min(-1) mg(-1)). To determine if NO could stimulate release of zinc and dimer dissociation, wild-type protein was allowed to react with an NO donor, DEA/NO, followed by buffer exchange. ICP-AES of samples treated with 10 microM DEA/NO showed a decrease in zinc content (0.23 +/- 0.01 to 0.09 +/- 0.01 mol of Zn/mol of iNOS dimer) with no loss of heme iron. Gel filtration of wild-type iNOS treated similarly resulted in approximately 20% more monomeric iNOS compared to a DEA-treated sample. Only wild-type iNOS had decreased activity (42 +/- 2%) after reaction with 50 microM DEA/NO compared to a control sample. Using the biotin switch method under the same conditions, only wild-type iNOS had increased levels of S-biotinylation. S-Biotinylation was mapped to C104 and C109 on wild-type iNOS using LysC digestion and MALDI-TOF/TOF MS. Immunoprecipitation of iNOS from the mouse macrophage cell line, RAW-264.7, and the biotin switch method were used to confirm endogenous S-nitrosation of iNOS. The data show that S-nitrosation of the zinc tetrathiolate cysteine results in zinc release from the dimer interface and formation of inactive monomers, suggesting that this mode of inhibition might occur in vivo.
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PMID:S-Nitrosation and regulation of inducible nitric oxide synthase. 1577 90

The determination of lead in tinplate by ICP-AES was presented in this paper. The tinplate samples were pretreated with NaOH-H2O2 for dissolving tin and lead,and the spectral interferences from iron could thus be avoided. The lead content in the sample was determined in the range of 0.14-0.38 microg/cm2. The recovery and the relative standard deviation (RSD) were in the ranges of 96.1%-103.2% and 2.6%-3.8%, respectively. The method was simple,rapid and efficient.
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PMID:[Determination of lead in tinplate by ICP-AES]. 1581 66

Concentrations of essential (copper, iron, manganese, selenium, and zinc) and non-essential (arsenic, cadmium, mercury, and lead) trace elements were measured in arms of Octopus vulgaris. The cephalopods were sampled from commercial fishery landings at two sites on the Portuguese coast in spring 2002 and 2003. Mercury was determined using an Advanced Mercury Analyser Spectrophotometer (AMAS) and other trace elements were measured using inductively coupled plasma-atomic emission spectrometry (ICP-AES). Since high levels of arsenic were detected, identification of the forms present was carried out using high performance liquid chromatography (HPLC) followed by ICP-MS. Mean concentrations of trace elements analysed were in the following order: As > Zn > Fe > Cu >> Cd > Pb > Se > Mn >> Hg. The results of speciation of arsenic demonstrate that virtually all arsenic was in the arsenobetaine form, which is the less toxic form. The concentrations of several trace elements were generally high in samples from Viana in 2002. Cadmium concentrations were above the legal limit for human consumption in samples from Viana in 2002 and two of these animals also had lead concentrations that exceeded legal limits. Mercury appeared in all samples but levels were within legally defined safe limits. No relationship was detected between trace element concentrations and size or maturity of octopus.
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PMID:Interannual patterns of variation in concentrations of trace elements in arms of Octopus vulgaris. 1583 85

The concentrations of cadmium, chromium, copper, iron, lead, manganese, nickel, and zinc in water and bed sediments of river Gomti have been studied in a fairly long stretch of 500 km from Neemsar to Jaunpur. Grab samples of water (October 2002-March 2003) and bed sediments (December 2002 and March 2003) were collected from 10 different locations following the standard methods. The river water and sediment samples were processed and analyzed for heavy metals viz., Cd, Cr, Cu, Fe, Pb, Mn, Ni, and Zn, and using ICP-AES. The heavy metals found in the river water were in the range: Cd (0.0001-0.0005 mg/L); Cr (0.0015-0.0688 mg/L); Cu (0.0013-0.0.0043 mg/L); Fe (0.0791-0.3190 mg/L); Mn (0.0038-0.0.0973 mg/L); Ni (0.0066-0.011 mg/L); Pb (0.0158-0.0276 mg/L); and Zn (0.0144-0.0298 mg/L) respectively. In the sediments the same were found in the range: Cd (0.70-7.90 /microg/g); Cr (6.105-20.595 microg/g); Cu (3.735-35.68 microg/g); Fe (5051.485-8291.485 micorg/g); Mn (134.915-320.45 microg/g); Ni (13.905-37.370 microg/g); Pb (21.25-92.15 microg/g); and Zn (1 5.72-99.35 microg/g) of dry weight respectively. Some physico-chemical parameters viz., pH, total solids, total dissolved solids, total suspended solids, dissolved oxygen, biological oxygen demand, chemical oxygen demand, hardness etc. were estimated as these have direct or indirect influence on the incidence, transport and speciation of the heavy metals. Based on the geoaccumulation indices, the Gomti river sediments from Neemsar to Jaunpur are considered to be unpolluted with respect to Cr, Cu, Fe, Mn, and Zn. It is unpolluted to moderately polluted with Pb. In case of Cd it varies from moderately polluted to highly polluted. As far as Ni is concerned the sediment is very highly polluted at Barabanki and Jaunpur D/s. No correlation was found between enrichment factor and geoaccumulation index.
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PMID:Status of heavy metals in water and bed sediments of river Gomti--a tributary of the Ganga River, India. 1595 11

Adsorption of Zn on goethite, hematite, birnessite, pyrolusite, corundum, and gibbsite was studied using a batch adsorption technique as a function of pH, zinc concentration in solution, and time of exposure. Adsorption from 0.01 M NaNO3 solutions undersaturated with respect to zinc (hydr)oxide at 3<pH<8 was found to be reversible and equilibrium was achieved in less than 24 h. A 2pK surface complexation model that assumes the constant capacitance of the electric double layer (CCM) and postulates the formation of positively charged >MeOZn+ complexes, where Me=Fe, Mn, and Al, was used to describe the dependence of adsorption equilibria on aqueous solution composition in a wide range of pH and Zn concentration. The logarithms of surface stability constant for Zn interaction with metal oxy(hydr)oxides (>MeOH0+Zn2+-->MeOZn+) vary from -2.5 to 0.5. They are higher for oxy(hydr)oxides than for anhydrous oxides. Stable isotopes of zinc in several filtrates were measured using an ICP-MS Neptune multicollector which made it possible, for the first time, to assess the degree of isotopic fractionation between 66Zn and 64Zn during zinc adsorption on mineral surfaces. The isotopic offset between aqueous solution and mineral surfaces (Delta(66/64)Zn(soln/solid)=delta((66/64)Zn)(solution)-delta((66/64)Zn)(surface)) was found to be weakly dependent on percentage of adsorbed metal and equals 0.20+/-0.03, 0.17+/-0.06, -0.10+/-0.03, -0.10+/-0.09, and -0.13+/-0.12 per thousand for goethite, birnessite, pyrolusite, corundum, and Al(OH)3. For hematite, Delta(66/64)Zn varies from -0.61+/-0.10 per thousand at pH 5.5 to -0.02+/-0.09 per thousand at 5.8<pH<6.7. Overall, zinc stable isotopic fractionation induced by adsorption on most mineral surfaces does not exceed 0.2 per thousand. We do not observe any correlation between the sign and magnitude of isotopic offset and the chemical nature of solid phase (hydrous versus anhydrous minerals), zinc surface adsorption constants (surface complexation model of the present work), and coordination and first-neighbor distances of surface >MeOZn(H2O)(n) complexes (available literature data on X-ray absorption spectroscopy). Apparently, the fine structure of surface complexes and the position and bond strength for second neighbors of zinc are likely to control its isotopic fractionation during adsorption on mineral surfaces. Our results strongly suggest that inorganic processes controlling zinc isotope adsorption on soil and sediment minerals should be of second-order importance compared to biological factors.
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PMID:Zinc stable isotope fractionation during its adsorption on oxides and hydroxides. 1596 23

Stable isotope absorption studies of iron have been limited by the high cost and limited availability of isotope ratio analysis using thermal ionization MS (TIMS). The development of high-resolution double focusing inductively coupled plasma MS (ICP-MS) may permit more cost-efficient sample analysis due to its high throughput, lower cost, easy sample pretreatment, and greater availability. Our objective was to develop an ICP-MS methodology for the measurement of iron isotope ratios using very small blood volumes. We developed a technique using multiple iron-nickel mixing standard solutions to adjust for nickel interference calibration. RBC samples from human subjects previously given 58Fe and 57Fe were analyzed for iron isotope ratios and compared with our current methodology (TIMS). Reproducibility of iron isotope ratios provided external relative SD < 0.5 and 0.7% (1 SD) for 57Fe/54Fe and 58Fe/54Fe, respectively. Iron isotope ratios from ICP-MS analysis did not differ from those from TIMS based on statistical analyses, nor did the calculated iron absorption values. The mean and SD of iron absorption did not differ when measured by TIMS or ICP-MS. A 2-microL RBC sample was sufficient for ICP-MS iron isotope ratio analysis with an internal relative SD < 0.5% and analytical time < 5 min. This technique may assist groups in increasing their use of stable isotope methods to assess iron absorption in infants and children.
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PMID:High resolution inductively coupled plasma mass spectrometry allows rapid assessment of iron absorption in infants and children. 1598 66

Two methods for separation of transferrin (Tf) sialoforms, capillary electrophoresis (CE) and high performance liquid chromatography (HPLC) with conventional UV absorbance detection, have been investigated and compared. First, conditions affecting the separation of the Tf isoforms by capillary zone electrophoresis and HPLC were carefully optimized. The use of 15 mmol L(-1) borate buffer (pH 8.4) containing 3 mmol L(-1) diaminobutane (DAB) as additive enabled good separation of the Tf isoforms by CE (75 cm x 50 microm i.d. fused silica capillary) at 25 kV. In HPLC, a gradient of ammonium acetate (from 0 to 250 mmol L(-1) in 45 min) buffered at pH 6 (Tris-HCl) proved suitable for separation of Tf isoforms on a Mono-Q HR 5/5 anion-exchange column. On-line specific detection of the iron associated with the different Tf isoforms, after Fe saturation, by inductively coupled plasma mass spectrometry (ICP-MS) was studied in detail to compare its analytical performance with UV detection. For both CE and HPLC an octapole reaction system (ORS) ICP-MS instrument was used to minimize polyatomic interferences on the (56)Fe major isotope. Limits of detection of the different isoforms were in the range of 0.02-0.04 micromol L(-1) Tf for HPLC-ICP (ORS)-MS. This hybrid technique proved more selective and reliable detection of transferrin isoforms with 2, 3, 4, 5, and 6 sialic acid residues (S(2), S(3), S(4), S(5), and S(6)) in real serum samples. Interesting results from iron speciation of Tf in serum from healthy individuals and from pregnant women are given.
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PMID:Detection of transferrin isoforms in human serum: comparison of UV and ICP-MS detection after CZE and HPLC separations. 1604 4

The combination of ion chromatography (IC) and inductively coupled plasma emission spectroscopy (ICP-ES) was used forthe sensitive and specific detection of hypophosphite (PO2), phosphite (PO3), methylphosphonic acid (MPA), and phosphate (PO4). Application of this technique to a wide range of environmental samples proved that reduced phosphorus was present in some situations including process water from thermal phosphorus plants, drinking water contacting cast iron, and phosphorus corrosion inhibitor used in water treatment and in sewage wastewater. Preliminary testing did not detect high concentrations of reduced phosphorus and phosphine in situations where it was previously reported to be very important, including anaerobic digesters in wastewater treatment plants. The new IC-ICP-ES technique is a promising tool for use in corrosion and soil research where phosphites are likely to be present.
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PMID:Analysis of reduced phosphorus in samples of environmental interest. 1604 69

Time courses of changes in manganese, iron, copper, and zinc concentrations were examined in regions of the brain of a 6-hydroxydopamine (6-OHDA)-induced rat model of Parkinson's disease using inductively coupled plasma mass spectrometry (ICP-MS). The concentrations were simultaneously determined in brain section at the level of the substantia nigra 1, 3, 7, 10, 14, and 21 days after the 6-OHDA treatment and compared with those of control rats. The distributions of these elements were obtained for 18 regions of the sagittal section (1-mm thick). The ICP-MS results indicated that Mn, Fe, Cu, and Zn levels of the 6-OHDA-induced parkinsonian brain were observed to increase in all regions that lay along the dopaminergic pathway. In the substantia nigra, the increase in Mn level occurred rapidly from 3 to 7 days and preceded those in the other elements, reaching a plateau in the 6-OHDA brain. Iron and Zn levels increased gradually until 7 days and then increased rapidly from 7 to 10 days. The increase in the copper level was slightly delayed. In other regions, such as the globus pallidus, putamen, and amygdala, the levels of Mn, Fe, Cu, and Zn increased with time after 6-OHDA treatment, although the time courses of their changes were region-specific. These findings contribute to our understanding of the roles of Mn and Fe in the induction of neurological symptoms and progressive loss of dopaminergic neurons in the development of Parkinson's disease. Manganese may hold the key to disturbing cellular Fe homeostasis and accelerating Fe levels, which play the most important role in the development of Parkinson's disease.
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PMID:Regional distributions of manganese, iron, copper, and zinc in the brains of 6-hydroxydopamine-induced parkinsonian rats. 1613 22

An on-line flow injection method for the direct determination of trace elements in environmental samples is described. A mini-column packed with 2,6-diacetylpyridine functionalized Amberlite XAD-4 was used to preconcentrate and separate 8 trace metals (Cd, Co, Cu, Mn, Ni, Pb, U and Zn) from water and extracts from solid samples. The metals were eluted with 0.1 M HNO(3) directly to the detection system (either inductively coupled plasma-mass spectrometry (ICP-MS) or flame atomic absorption spectrometry (FAAS)). As well as demonstrating that the resin could be used to preconcentrate ultra-trace analytes from natural waters, it was also shown to work well at a pH of 5.5. Therefore, after treatment of sample digests with sodium fluoride, samples that contain extremely large concentrations of iron may be analysed for trace analytes without the excess iron overloading the capacity of the resin. To this end, the analytes Cd, Co, Cu and Ni were preconcentrated from acid extracts of certified soil/sediment samples and then eluted with nitric acid to be determined on-line. Limits of detection (3sigma) of Cd = 0.33 microg l(-1), Co = 0.094 microg l(-1), Cu = 0.34 microg l(-1), Mn = 0.32 microg l(-1), Ni = 0.30 microg l(-1), Pb = 0.43 microg l(-1), U = 0.067 microg l(-1) and Zn = 0.20 microg l(-1) for the FI-ICP-MS system and Cd = 22 microg l(-1), Co = 60 microg l(-1), Cu = 10 microg l(-1) and Ni = 4.8 microg l(-1) for the FI-FAAS system were obtained. Analysis of certified reference materials showed good agreement with the certified values using the two methods.
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PMID:Preconcentration and determination of trace elements with 2,6-diacetylpyridine functionalized Amberlite XAD-4 by flow injection and atomic spectroscopy. 1622 74


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