Effect of carbon cup aging on plasma zinc determination by flameless atomic adosorption spectrometry.
(73/1258)
Determination of zinc in blood plasma by flameless atomic absorption spectrometry is discussed, with particular reference to the protocol required for the successful use of the Varian-Techtron Carbon Rod Atomizer. Cup aging is shown to be an important factor in limiting the precision of this analytical technique and ways of minimizing the problem are described. Matrix problems have also been encountered, which precluded the use of aqueous standard curves and the method of standard additions. We propose the use of plasma in preparing standard curves, the values for which are corrected for inherent plasma zinc, as a possible solution to the problem. (+info)
Structural characterization and thermal stability of Notothenia coriiceps metallothionein.
(74/1258)
Fish and mammalian metallothioneins (MTs) differ in the amino acid residues placed between their conserved cysteines. We have expressed the MT of an Antarctic fish, Notothenia coriiceps, and characterized it by means of multinuclear NMR spectroscopy. Overall, the architecture of the fish MT is very similar to that of mammalian MTs. However, NMR spectroscopy shows that the dynamic behaviour of the two domains is markedly different. With the aid of absorption and CD spectroscopies, we studied the conformational and electronic features of fish and mouse recombinant Cd-MT and the changes produced in these proteins by heating. When the temperature was increased from 20 to 90 degrees C, the Cd-thiolate chromophore absorbance at 254 nm of mouse MT was not modified up to 60 degrees C, whereas the absorbance of fish MT decreased significantly starting from 30 degrees C. The CD spectra also changed quite considerably with temperature, with a gradual decrease of the positive band at 260 nm that was more pronounced for fish than for mouse MT. The differential effect of temperature on fish and mouse MTs may reflect a different stability of metal-thiolate clusters of the two proteins. Such a conclusion is also corroborated by results showing differences in metal mobility between fish and mouse Zn-MT. (+info)
Spectral and metal-binding properties of three single-point tryptophan mutants of the human transferrin N-lobe.
(75/1258)
Human serum transferrin N-lobe (hTF/2N) contains three conserved tryptophan residues, Trp(8), Trp(128) and Trp(264), located in three different environments. The present report addresses the different contributions of the three tryptophan residues to the UV-visible, fluorescence and NMR spectra of hTF/2N and the effect of the mutations at each tryptophan residue on the iron-binding properties of the protein. Trp(8) resides in a hydrophobic box containing a cluster of three phenylalanine side chains and is H bonded through the indole N to an adjacent water cluster lying between two beta-sheets containing Trp(8) and Lys(296) respectively. The fluorescence of Trp(8) may be quenched by the benzene rings. The apparent increase in the rate of iron release from the Trp(8)-->Tyr mutant could be due to the interference of the mutation with the H-bond linkage resulting in an effect on the second shell network. The partial quenching in the fluorescence of Trp(128) results from the nearby His(119) residue. Difference-fluorescence spectra reveal that any protein containing Trp(128) shows a blue shift upon binding metal ion, and the NMR signal of Trp(128) broadens out and disappears upon the binding of paramagnetic metals to the protein. These data imply that Trp(128) is a major fluorescent and NMR reporter group for metal binding, and possibly for cleft closure in hTF/2N. Trp(264) is located on the surface of the protein and does not connect to any functional residues. This explains the facts that Trp(264) is the major contributor to both the absorbance and fluorescence spectra, has a strong NMR signal and the mutation at Trp(264) has little effect on the iron-binding and release behaviours of the protein. (+info)
Glucose-induced activation of rubidium transport and water flux in sunflower root systems.
(76/1258)
Excised 20-d-old sunflower roots (Helianthus annuus L. cv. Sun-Gro 393) were used to study the effect of different sugars on rubidium and water fluxes. The roots sensed and absorbed glucose from the external medium inducing the activation of rubidium accumulated in the root (Rb(+) root), the flux of exuded rubidium (J(Rb)) and, to a lesser degree, the exudation rate (J(v)). These effects were also triggered by fructose, but not by 6-deoxyglucose (6-dG), a glucose analogue which is not a substrate for hexokinase (HXK). The effect of 2-deoxyglucose (2-dG), an analogue that is phosphorylated but not further metabolized, was complex, suggesting an inhibitory effect on solute transport to the xylem. The amounts of glucose required to activate rubidium and water fluxes were similar to those previously reported to regulate different processes in other plants (0.5--10 mM). When sorbitol was used instead of glucose, neither rubidium uptake (Rb(+) root plus J(Rb)) nor J(v) was activated. It is proposed that glucose present in the root plays an important signalling role in the regulation of Rb(+) (K(+)) and water transport in plant roots. (+info)
Iron concentrations in human dermis assessed by microdialysis associated with atomic absorption spectrometry.
(77/1258)
Until recently, the determination of metallic elements concentrations in normal skin, in vivo, was rare due to the lack of non-invasive techniques. Microdialysis has the advantage of being slightly invasive when applied to the collection in vivo of endogenous or exogenous substances from the skin. Iron is an active element in different cutaneous disorders. The aim of this work was to assess iron by atomic absorption spectrometry (AAS) after the collection of samples by microdialysis from human dermis. A first essential step, before determining the in vivo iron concentration in human dermis, was to establish an experimental protocol applicable to ex vivo as well as in vivo conditions. For this reason, this work deals only with the assessment of iron in ex vivo human dermis. A skin microdialysis technique and a calibration method, the No Net Flux, were used to quantify basal iron concentrations in human dermis and the same method was also used to determine in vitro and ex vivo iron recoveries. No differences were detected between in vitro and ex vivo recoveries. Ex vivo basal iron dermis concentrations ranged from 3.6 to 7.7 microg/l. This study shows that non-invasive microdialysis is an efficient method for sampling iron from human skin. A sensitive and accurate AAS technique was able to assess low iron concentrations in human dermis. The strategy adopted for this work was efficient and appropriate for the determination of iron in human skin and experiments will be carried out in vivo. (+info)
Hepatic iron storage in very low birthweight infants after multiple blood transfusions.
(78/1258)
OBJECTIVE: To investigate the effect of multiple blood transfusions on hepatic iron storage in preterm, very low birthweight (VLBW) infants. METHODS: Seventeen VLBW infants who died within the first six months of life and underwent postmortem examination were studied. Serum ferritin, iron, and total iron binding capacity were measured within the week before the infants' death. Liver iron concentration was quantitatively determined by atomic absorption spectrophotometry and semiquantitatively assessed by histochemical liver iron grading. The clinical characteristics and the iron results were compared between infants receiving < 100 ml of blood (group A) and those receiving >/= 100 ml (group B). Spearman's correlation coefficient was used to evaluate the relation between the volume of blood transfused and serum/liver iron concentrations. Statistically significant variables associated with liver iron concentration were further subjected to multivariate stepwise regression analysis. RESULTS: Infants in group B had significantly higher serum iron (p < 0.01), serum ferritin (p < 0.01), and liver iron concentration (p < 0.01) than those in group A. The total and net volume of blood transfused were significantly associated with liver iron concentration (p < 0.001, r = 0.86; p < 0.001, r = 0.71 respectively), semiquantitative histochemical liver iron grading (p < 0.001, r = 0.80; p < 0.005, r = 0.71 respectively), and serum ferritin (p < 0.001, r = 0.84; p < 0.01, r = 0.69 respectively). In addition, both liver iron concentration and liver iron grading were found to be significantly associated with serum ferritin (p < 0.001, r = 0.76; p < 0.005, r = 0.68 respectively). Multivariate stepwise regression analysis indicated that the (log) liver iron concentration was significantly associated with the (log) volume of blood transfusion (p < 0.001; regression coefficient 0.39, SE 0.09), after adjustment for gestational age (R(2) = 0.84). CONCLUSIONS: This study showed a significant positive relation between the volume of blood transfused and the liver iron concentration in preterm VLBW infants. Although the transfusional blood volume correlated closely with the amount of iron deposited in hepatic tissues, clinical manifestations of iron overload were not observed. Carers should be aware of this potential harmful effect before prescribing blood or routine iron supplement to vulnerable preterm infants. (+info)
Corrosion characteristics of ferric and austenitic stainless steels for dental magnetic attachment.
(79/1258)
The corrosion behaviors of four ferric stainless steels and two austenitic stainless steels were examined in a simulated physiological environment (0.9% NaCl solution) to obtain basic data for evaluating the appropriate composition of stainless steels for dental magnetic attachments. The corrosion resistance was evaluated by electrochemical techniques and the analysis of released metal ions by atomic absorption spectrophotometry. The surface of the stainless steels was analyzed by X-ray photoelectron spectroscopy (XPS). The breakdown potential of ferric stainless steels increased and the total amount of released metal ions decreased linearly with increases in the sum of the Cr and Mo contents. The corrosion rate of the ferric stainless steels increased 2 to 6 times when they were galvanically coupled with noble metal alloys but decreased when coupled with commercially pure Ti. For austenitic stainless steels, the breakdown potential of high N-bearing stainless steel was approximately 500 mV higher than that of SUS316L, which is currently used as a component in dental magnetic attachments. The enriched nitrogen at the alloy/passive film interface may be effective in improving the localized corrosion resistance. (+info)
Retention time of macerated alfalfa hay and silage in sheep.
(80/1258)
Fresh alfalfa was mowed and conditioned mechanically at four levels: a control (rubber rolls), macerated once (a single passage through three finely corrugated rolls set at 1-mm clearance), macerated twice (two passages), and macerated thrice (three passages). Alfalfa was then field-wilted either for 45 h and conserved as chopped silage at 30% dry matter (DM) or for 94 h and stored as baled hay at 85% DM. The eight forage treatments (four mechanical conditioning levels x two conservation systems) were fed to 24 sheep (three replications per treatment) during 5 wk. At the beginning of wk 5, a 15-g sample of chromium-mordanted forage (3.5% Cr) was fed to each sheep, and feces samples were collected at 30 different times over 7 d, between 10 h and 168 h after Cr ingestion. Four models were used to estimate the passage rates, the time delay, and the mean retention time (MRT). A two-compartment time-dependent model and a multicompartment model produced the best fit (average r2 of 0.96) to represent the Cr concentration in the feces over time. When compared with alfalfa hay, alfalfa silage had a higher (P < 0.01) time-dependent turnover rate (0.0949 vs 0.0733/h), a lower (P = 0.03) time delay (9.1 vs 11 h), and a lower (P = 0.04) MRT (57.8 vs 64.4 h). Maceration did not affect significantly (P > 0.10) the time delay or the MRT. However, the MRT of macerated alfalfa hay tended to be higher than the MRT of control hay. Experimental data based on marker concentration in the feces can be used satisfactorily to assess differences in MRT between treatments, but they should be used with caution to estimate the partition of retention time within the gastrointestinal tract. (+info)