Trace element requirements, intake and recommendations.
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Biomedical subjects
Publications and source records attributed to G V Iyengar.
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Over 100 years ago, macro-scale analytical techniques were used to discover the roles of special compounds (especially of metallic elements) in living organisms, and investigations were focussed on selected proteins and pigments suspected of containing percentage quantities of metals. In contrast, present-day analytical techniques are capable of detecting extremely small quantities and have become routine ultra-trace measurement tools to probe elemental interactions at cellular levels. The scientific achievements connecting these two boundaries are punctuated with an array of analytical developments; some highlighting the phenomenal advances in the measurement technology and others reflecting the exceptional bioanalytical perception and the multi-disciplinary outlook of trace element investigators. An account of the events that contributed to the overall progress in biological trace element research is the essence of this communication.
There has been considerable progress in understanding the role of chromium in human nutrition. However, the lack of data on the forms of chromium-absorption from foods by the gastro-intestinal tract, and our concomitant inability to obtain an accurate assessment of the daily mobile pool of metabolically active chromium in the human body continues to be an impediment in assessing the overall impact of chromium nutrition. Based on recent chromium data on human tissues and body fluids available in the literature, an assessment of chromium content in different body compartments as well as in the total body is presented. However, on the analytical side, a few problems still persist and, therefore, a reliable analysis for chromium is restricted to a few selected laboratories.
Using 201 foods from the United States Food and Drug Administration's Total Diet Study (FDA TDS), a mixed diet composite (USDIET-I) was prepared to represent the intake of 25-30-year-old males in the United States. Proximate analyses, phytate determination, and assays for nutrient elements and selected toxic elements, as well as organic nutrients were carried out on this composite. As part of a quality control exercise for a coordinated research program, atomic absorption spectrophotometry, inductively coupled atomic emission spectrometry, colorimetry and neutron activation analysis were used to determine up to 30 elements in this diet material. A comparison of the daily intakes of As, Ca, Cd, Cu, Fe, Hg, K, Mg, Mn, Na, P, Se and Zn from the composite USDIET-I shows excellent to good agreement with FDA TDS values calculated from results of single food analyses. These USDIET-I results demonstrate the feasibility of the mixed diet concept as a viable approach for a reliable assessment of daily intakes, especially for a number of elements such as Cd, Cr, Hg and Mo that occur at low concentrations in individual food products. Simultaneously, stability of some organic nutrients during storage was also investigated. Initial findings suggest that this program may also be useful in the development of reference materials for organic nutrients, for which there is a great need. These aspects are discussed.
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Over 60 investigators, some of them with an outstanding international reputation as analysts, were requested to identify reliable data from their countries for elemental concentrations in normal adult human samples of clinical interest by contributing their own data as well as screening the literature information. A set of samples consisting of whole blood and its components, urine, milk, liver and hair were chosen and considered for 15 elements of biological significance: Zn, Cu, Fe, Se, Mn, Pb, Cd, Hg, As, Mo, Cr, Co, I, Ni and F. The results partly cover over 40 countries from the global regions of Africa, Asia, Europe, North, South and Central America, Australia and New Zealand. This survey has been useful in demonstrating certain trends of trace element picture around the world, at least qualitatively. Both diet and environment have a strong influence on the distribution pattern of several elements such as As, Cd, Mn, Pb, Se and Zn. A limited comparison of the available information on soil status of different countries revealed interesting associations for elements such as Mn and Zn. Importantly, this study revealed that only a few countries were in a position to supply a reasonable amount of data on samples requested here. In particular, for a number of countries, reliable data for even very essential elements such as Cu, Zn and Fe were not available. In view of the nutritional importance of several elements the time is ripe for international organizations to intervene and help produce some reference data for selected global regions which lack data of any kind.
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Concentrations of Co, Cu, Fe, Hg, Mn, Sb, Se and Zn in IAEA milk (dry) standard A-11 were re-evaluated with the help of instrumental and radiochemical neutron activation analysis (NAA). The results show reasonably good agreement for Co (5.1 +/- 0.55 ng/g) and Zn (34 +/- 2.5 micrograms/g), in relation to the recommended values. For Cu (374 +/- 15 ng/g), Fe (2.4 +/- 0.34 microgram/g) and Mn (250 +/- 20 ng/g); the results obtained are lower than the reported values. Cu, Mn, and Zn were cross-checked by atomic absorption spectrophotometry (AAS). The results, Cu = 331 +/- 27 ng/g, Mn = 302 +/- 62 ng/g, and Zn = 35 +/- 1 microgram/g, fall within the range of mean values obtained by NAA. for Hg and Se, using instrumental NAA, only upper limits could be indicated; because of the low levels of concentrations of Hg and Se on one hand, and high content of P (9100 micrograms/g) in A-11 milk standard on the other, NAA coupled with radiochemistry is to be preferred for these two elements. In pooled human milk, in addition to Cu and Mn (radiochemical) and Co, Fe, Sb and Zn (instrumental), Hg and Se could also be determined non-destructively because of the favorable Hg/P and Se/P ratios in this matrix.
Scalp hair, fingernail and water samples collected from different parts of Egypt are analysed by atomic absorption spectrophotometry and neutron activation analysis. The results for hair show minimum variation of Zn among different regions; a relatively well-controlled dispersion of values (maximum to minimum less than 2) for Co, Fe, Mn and Se; an overall inter-regional variation of factors of 7, 10, 6, 4 and 6 for the elements Ag, Cd, Cs, Sc and W, respectively; a steady decline in the concentration of Sc from south (Aswan) to north (Alexandria). The findings for nail show a steady decline in the concentration of Fe and Sc from south (Aswan) to north (Alexandria); elevated levels of Cd in samples from Aswan area; and regional variations extending up to factors of 6.5, 3.6, 4.7, 5.9, 4.4, 4.5 and 1.9 for Co, Cr, Cs, Mn, Mo, W and Zn, respectively. No unified relationship is observed between the elemental compositions of hair and nail. For Cr, Cs, Fe, Sb and Sc mean values for hair are lower than in nails. Among the remaining elements the ratio nail/hair is less than 1 for Ag in Assiut, El Kharga and Alexandria, for Cd in all of the areas with the exception of Aswan, for Co, Mn and Mo in Cairo, for Zn in Cairo and Alexandria, and for W in Alexandria. The data for water samples reflect highest concentration of all the measured elements in the Mediterranean sea; a steady increase of the concentration of Cu in drinking water from south to north and for Nile water a similar trend for Ca and Mg; and elevated concentrations of Sb and Mn in river water. No distinct trend for the interrelationship between water and tissue elemental concentrations could be established for the regions around Cairo, Aswan and Alexandria.
The activity of the selenoenzyme glutathione peroxidase (EC 1.11.1.9) was determined in platelets of 15 patients with acute myocardial infarction and 13 control subjects. The platelets of the patients had significantly lower activities of the enzyme (P(t) greater than 0.99). This may be related to the pathogenesis of the disease.
The implication of post-mortem changes such as cell swelling, imbibition and autolysis on the elemental composition of body organs has been studied in rats. Liver has been chosen as an example. Retaining the liver inside the intact dead body for different periods of time at ambient temperature induced significant changes in its weight due to post-mortem tissue degeneration. Livers from animals that were frozen at -15 degrees C also showed significant decreases in weight when they were thawed on the third day. The effect of these changes on the concentrations of various elements depended on the association of the elements with extracellular fluid and intracellular components. For example, concentration of K+ was affected more by the lysis of the cell and sustained losses up to 30 per cent. in relation to the control values, while the total content was reduced by more than 40 per cent. as a result of both lysis and tissue liquefaction. For Na+, differences ranging from +10 to -20 per cent. in concentrations and +20 to -40 per cent. in total content were observed which could be explained by the movement of fluid in and out of the organ. Among trace elements, variation observed for iron ranged from -20 to +40 per cent., while both copper and zinc were found to fluctuate between -20 and +20 per cent. The loss observed in the total content in the liver for the five trace elements studied (Cu, Fe, Mn, Rb and Zn) was found to be about 20 to 40 per cent.
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The differences in concentrations of cesium, cobalt, iron, mercury, rubidium, selenium and zinc between serum and plasma were examined with the aid of instrumental neutron activation analysis. Eighty serum and plasma samples obtained from 13 donors were compared. Serum was prepared in plastic tubes immediately after clotting, and plasma was separated with heparin as anticoagulant. No significant differences in the concentrations of cesium, cobalt, mercury and selenium were observed. However, the concentrations of iron, rubidium and zinc were significantly higher in serum than in plasma. The average differences were 322, 12 and 20 ng/ml for iron rubidium and zinc, respectively. The average differences found for cesium, rubidium and zinc were far below that which can be expected from a complete, or considerable release of these elements from platelets which aggregate or disintegrate during the clotting process in preparing serum.
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Sampling of platelets for trace-element analysis poses special problems: obtaining adequate sample material, achieving a sufficient cell purity, preserving viability (integrity), correcting for trapped plasma, and controlling contamination. We used a blood-cell separator for the primary isolation of platelets from blood, and differential centrifugation in natural plasma to further isolate them. The pyrimidopyrimidine RA233 was used as a stabilizer to maintain viability. 131I-labeled human serum albumin was used to estimate trapped plasma. Contamination was controlled by using five-times-distilled water to simulate donor's blood in the system and by comparing three fractions: the serum, the first portion of the platelet-rich plasma, and the supernatant plasma after the final centrifugation. Neutron activation analysis was used for the elemental analysis. A single differential centrifugation of the platelet-rich plasma from the blood-cell separator at 400 x g for 8 min was optimum (mean mass fractions:erythrocytes/platelets less than 5 mg/g and leukocytes/platelets less than 20 mg/g). The trapped plasma in the wet platelet samples amounted to about 0.40 g/g. No appreciable contamination from the sampling system was found for the elements Ag, Cd, Co, Cr, Cs, Cu, Fe, Mo, Rb, Sb, Se, and Zn.
We determined the elements Cu, Fe, K, and Zn in normal human platelets by neutron activation analysis. The platelets were obtained from seven donors and treated as described in Part I. The elemental composition is expressed on a wet-weight basis for plasma-free platelets. The following results were obtained (+/- values are 1 SD); "Pure" platelets: trapped plasma = 378 +/- 35 mg/g, water content = 715 +/- 15 mg/g, mean weight of the single platelet (by two different methods) = 9.9 +/- 1.1 pg and 11.2 +/- 1.7 pg, K = 4.39 +/- 1.06 mg/g, Zn = 49.23 +/- 10.97 microgram/g, Fe = 12.28 +/- 2.94 microgram/g, and Cu = 1.39 +/- 0.25 microgram/g. "Impure" platelets: trapped plasma = 349 +/- 31 mg/g, water content = 736 +/- 12 mg/g, K = 3.26 +/- 0.78 mg/g, Zn = 35.71 +/- 7.99 microgram/g, Fe = 17.11 +/- 5.10 microgram/g, and Cu = 1.39 +/- 0.21 microgram/g. To our knowledge, no data on Fe and Cu in platelets have hitherto been reported.
The elements Ag, Au, Cd, Co, Cr, Cs, Mo, Rb, Sb, and Se were determined in platelets from seven normal donors. The results, in ng/g wet weight, for plasma-free platelets follow: "Pure" platelets: Ag = 29 +/- (18), au = 0.22 +/- (0.22), Cd = 6.2 +/- 3.4, Cs = 54.8 +/- 19.2, Cr = 6.1 +/- 2.5, Co = 7.5 +/- (5.0), Mo = 3.4 +/- 1.3, Rb = 10400 +/- 3000, Sb = 18 +/- (26), and Se = 782 +/- 127. "Impure" platelets: Au = 0.23 +/- (0.28), Cd = 6.4 +/- 2.6, Cs = 35.2 +/- 13.8, Cr = 8.2 +/- 2.9, Co = 2.9 +/- (3.0), Mo = 3.2 +/- 0.8, Rb = 8700 +/- 1700, Sb = 13.2 +/- (8.7), and Se = 679 +/- 57. To our knowledge, none of these 10 trace elements has been determined in platelets before. The selenium concentration in platelets exceeds that in other tissues (e.g., liver). We suggest that glutathione peroxidase or other unknown selenoenzymes are particularly important in platelet metabolism. Platelets are crucial for triggering thrombosis, and so may be involved as links between selenium deficiency and the concomitant increased death rate from cardiovascular disease.