Reference Intervals of Hematology and Clinical Chemistry Analytes for 1-Year-Old Korean Children.

BACKGROUND: Reference intervals need to be established according to age. We established reference intervals of hematology and chemistry from community-based healthy 1-yr-old children and analyzed their iron status according to the feeding methods during the first six months after birth. METHODS: A total of 887 children who received a medical check-up between 2010 and 2014 at Boramae Hospital (Seoul, Korea) were enrolled. A total of 534 children (247 boys and 287 girls) were enrolled as reference individuals after the exclusion of data obtained from children with suspected iron deficiency. Hematology and clinical chemistry analytes were measured, and the reference value of each analyte was estimated by using parametric (mean±2 SD) or nonparametric methods (2.5-97.5th percentile). Iron, total iron-binding capacity, and ferritin were measured, and transferrin saturation was calculated. RESULTS: As there were no differences in the mean values between boys and girls, we established the reference intervals for 1-yr-old children regardless of sex. The analysis of serum iron status according to feeding methods during the first six months revealed higher iron, ferritin, and transferrin saturation levels in children exclusively or mainly fed formula than in children exclusively or mainly fed breast milk. CONCLUSIONS: We established reference intervals of hematology and clinical chemistry analytes from community-based healthy children at one year of age. These reference intervals will be useful for interpreting results of medical check-ups at one year of age.

Stability of Vitamin A, Iron and Zinc in Fortified Rice during Storage and Its Impact on Future National Standards and Programs--Case Study in Cambodia.

Fortified rice holds great potential for bringing essential micronutrients to a large part of the world population. The present study quantified the losses of three different micronutrients (vitamin A, iron, zinc) in fortified rice that were produced using three different techniques (hot extrusion, cold extrusion, and coating) and stored at two different environments (25 ± 5 °C at a humidity of 60% and 40 ± 5 °C at a humidity of 75%) for up to one year. Fortified rice premix from the different techniques was mixed with normal rice in a 1:100 ratio. Each sample was analyzed in triplicate. The study confirmed the high stability of iron and zinc during storage while the retention of vitamin A was significantly affected by storage and the type of techniques used to make rice premix. Losses for iron and zinc were typically <10% for any type of rice premix. After 12 months at mild conditions (25 °C and humidity of 60%), losses for vitamin A ranged from 20% for cold extrusion, 30% for hot extruded rice 77% for coated rice premix. At higher temperatures and humidity, losses of vitamin A were 40%-50% for extruded premix and 93% for coated premix after 6 months. We conclude that storage does lead to a major loss of vitamin A and question whether rice is a suitable food vehicle to fortify with vitamin A. For Cambodia, fortification of rice with iron and zinc could be an effective strategy to improve the micronutrient status of the population if no other food vehicles are available.

Population-based pediatric reference intervals for hematology, iron and transferrin.

Reference intervals are crucial decision-making tools aiding clinicians in differentiating between healthy and diseased populations. However, for children such values often are lacking or incomplete. Blood samples were obtained from 689 healthy children, aged 6 months to 18 years, recruited in day care centers and schools. Hematology and anemia analytes were measured on the Siemens Advia 2120 and Abbott Architect ci8200 platforms (hemoglobin, erythrocyte volume fraction [EVF], erythrocytes, mean corpuscular volume [MCV], mean corpuscular hemoglobin [MCH], mean corpuscular hemoglobin concentration [MCHC], reticulocytes, leukocytes, lymphocytes, monocytes, neutrophils, eosinophils, basophils, platelets, iron, transferrin, transferrin saturation). Age- and gender-specific pediatric reference intervals were defined by calculating 2.5th and 97.5th percentiles. The data generated is primarily applicable to a Caucasian population, but could be used by any laboratory if verified for the local patient population.

Development of a certified reference material (NMIJ CRM 7531-a) for the determination of trace cadmium and other elements in brown rice flour.

A certified reference material (CRM) for trace cadmium and other elements in brown rice flour was developed at the National Metrology Institute of Japan (NMIJ). The CRM was provided as a dry powder after drying and frozen pulverization of fresh brown rice obtained from a Japanese domestic market. Characterization of the property value for each element was carried out exclusively by NMIJ with at least two independent analytical methods, including inductively coupled plasma mass spectrometry (ICP-MS), ICP high-resolution mass spectrometry, isotope-dilution ICP-MS, ICP optical emission spectrometry, and graphite-furnace atomic-absorption spectrometry. Property values were provided for six elements (Mn, Fe, Cu, Zn, As, and Cd). The concentration range of the property values was from 0.280 mg kg(-1) of As to 31.8 mg kg(-1) of Zn. The combined relative standard uncertainties of the property values were estimated by considering the uncertainties of the homogeneity, characterization, difference among analytical methods, dry-mass correction factor, and calibration standard. The range of the relative combined standard uncertainties was from 1.1% of Zn to 1.6% of As.

Arsenic-enriched groundwaters of India, Bangladesh and Taiwan--comparison of hydrochemical characteristics and mobility constraints.

Arsenic (As) enrichment in groundwater has become a major global environmental disaster. Groundwater samples were collected from 64 sites located in the districts of 24-Parganas (S), and Nadia in West Bengal, India (Bhagirathi sub-basin), and 51 sites located in the districts of Comilla, Noakhali, Magura, Brahman baria, Laxmipur, Munshiganj, Faridpur and Jhenaida in Bangladesh (Padma-Meghna sub-basin). Groundwater samples were also collected from two As-affected areas (Chianan and Lanyang plains) of Taiwan (n = 26). The concentrations of major solutes in groundwater of the Padma-Meghna sub-basin are more variable than the Bhagirathi sub-basin, suggesting variations in the depositional and hydrological settings. Arsenic concentrations in groundwaters of the studied areas showed large variations, with mean As concentrations of 125 µg/L (range: 0.20 to 1,301 µg/L) in Bhagirathi sub-basin, 145 µg/L (range: 0.20 to 891 µg/L) in Padma-Meghna sub-basin, 209 µg/L (range: 1.3 to 575 µg/L) in Chianan plain, and 102 µg/L (range: 2.5 to 348 µg/L) in Lanyang plain groundwater. The concentrations of Fe, and Mn are also highly variable, and are mostly above the WHO-recommended guideline values and local (Indian and Bangladeshi) drinking water standard. Piper plot shows that groundwaters of both Bhagirathi and Padma-Meghna sub-basins are of Ca-HCO(3) type. The Chianan plain groundwaters are of Na-Cl type, suggesting seawater intrusion, whereas Lanyang plain groundwaters are mostly of Na-HCO(3) type. The study shows that reductive dissolution of Fe(III)-oxyhydroxides is the dominant geochemical process releasing As from sediment to groundwater in all studied areas.

Magnetic labeling of non-phagocytic adherent cells with iron oxide nanoparticles: a comprehensive study.

Small particles of iron oxide (SPIO) and ultrasmall particles of iron oxide (USPIO), inducing a strong negative contrast on T(2) and T(2)*-weighted MR images, are the most commonly used systems for the magnetic labeling of cultured cells and their subsequent detection by magnetic resonance imaging (MRI). The purpose of this work is to study the influence of iron incubation concentration, nanoparticle size and nanoparticle coating on the magnetic labeling and the viability of non-phagocytic adherent cells in culture. The magnetic labeling of 3T6 fibroblasts was studied by T(2)-weighted MRI at 4.7 T and by dosing-or cytochemical revealing-of iron through methods based on Perl's Prussian blue staining. Cells were incubated for 48 h with increasing iron concentrations of SPIO (25-1000 microg Fe/ml Endorem. Sinerem, a USPIO (20-40 nm) coated with neutral dextran, and Resovist (65 nm), a SPIO bearing an anionic carboxydextran coating, were compared with Endorem (dextran-coated, 80-150 nm) as magnetic tags. The iron loading of marrow stromal cell primary cultures (MSCs) isolated from rat femurs was compared with that of 3T6 fibroblasts. The SPIO-labeling of cells with Endorem was found to be dependent on the iron incubation concentration. MSCs, more sparsely distributed in the culture, exhibited higher iron contents than more densely populated 3T6 fibroblast cultures. A larger iron loading was achieved with Resovist than with Endorem, which in turn was more efficient than Sinerem as a magnetic tag. The magnetic labeling of cultured non-phagocytic adherent cells with iron oxide nanoparticles was thus found to be dependent on the relative concentration of the magnetic tag and of the cells in culture, on the nanoparticle size, and on the coating type. The viability of cells, estimated by methods assessing cell membrane permeability, was not affected by magnetic labeling in the conditions used in this work.

Contents of bioelements and toxic metals in the Polish population determined by hair analysis. Part III. Adults aged 20 to 40 years.

The aim of the present study was to define referential values of 5 basic bioelements (Ca, Mg, Zn, Cu, Fe) and 2 toxic metals (Pb and Cd) in the hair of Polish adults aged 20 to 40 years, based on the research conducted from 1991 to 2006 on a group of 5733 formally healthy subjects of both sexes. The results of our biochemical tests were subjected to statistical analysis and they can be treated as referential norms for the concentrations of analyzed bioelements in the hair of the Polish population. Our analyses confirm significant differences in accumulations of certain elements in women's and men's hair. We established that women's hair Ca, Mg, Zn (p = 0.000000) and hair Cu (p = 0.0001) concentrations are higher than they are in men. A lower mean of hair Fe concentrations in women were confirmed by some tests. Men have significantly higher (p = 0.0000) concentrations of toxic metals (hair Pb and Cd), which keep increasing with age. Analyses of correlations confirm significance (for the assumed level of significance alpha = 0.05) of synergistic interactions between bioelements: Ca-Mg (r = +0.70), Ca-Zn (r = +0.39), Ca-Cu (r = +0.14), Mg-Zn (r = +0.31), Mg-Cu (r = +0.10), Zn-Cu (r = +0.11), Zn-Fe (r = +0.07). Significance (a < 0.05) of antagonistic interactions with toxic metals: Ca-Pb (r = -0.13), Zn-Pb (r = -0.15), Mg-Pb (r = -0.15), Zn-Cd (r =-0.02) was confirmed. Significance (alpha < 0.05) of synergistic effects with toxic metals: Pb-Cd (r = +0.35), Fe-Pb (r = +0.16), Fe-Cd (r = +0.08) was also confirmed. Adults in whose hair bioelements and hair toxic metals concentrations have shown values outlying from reference and who have too many toxic elements and too few bioelements should undergo further diagnostic tests since the results could be a sign of disturbances that could lead to various diseases.

[Determination of trace lead and iron in nickel chloride and manganese sulfate by flame atomic absorption spectrometry after coprecipitation with yttrium phosphate].

Using yttrium phosphate as the coprecipitation collector for the separation and preconcentration of trace lead and iron in nickel chloride and manganese sulfate, flame atomic absorption spectrometric (FAAS) determination was described in the present paper. Coprecipitation parameters including the pH of the solution, and the amounts of YCl3 and H3 PO4 were discussed. It was found that lead and iron in nickel chloride could be coprecipitated quantitatively in the range of pH 3.0-4.0, and so could be lead in manganese sulfate. The detection limits (3sigma) of lead and iron in 20 mL solution were 1.63 x 10(-2) mg x L(-1) and 4.58 x 10(-2) mg x L(-1) respectively. In NiCl2 solution the standard addition recoveries for lead and iron were 100.91% and 99.73% respectively, and in MnSO4 solution the standard addition recoveries were 99.45% and 98.98% respectively. The method has eliminated the interference of matrix, and the result is satisfied.

Determination of Iron in Caco-2 cells by ET-AAS.

An electrothermal atomic absorption spectrometric method (ET-AAS) was developed for the direct determination of iron in intestinal Caco-2 cells after studying cell viability and proliferation using 3-(4,5-dimethylthiazole-2-yl)-2,5-diphenyl tetrazolium bromide (MTT test). Zeeman background correction and end-capped graphite tubes with L'vov platforms were used. Samples were dissolved in dimethylsulfoxide (DMSO) and pipetted directly into the graphite tube. The preashing, pretreatment and atomization steps were optimized. The temperatures selected were 600, 1200, and 2100 degrees C, respectively. Stability measurements were performed using iron standard solutions in DMSO on the one hand and acidified cell solutions on the other. Direct measurement and standard addition were compared in order to determine possible influences of the matrix. The low detection limit of the ET-AAS method (1.3 microg/L or 3.3 microg/g) combined with the small sample quantities required are ideal for the determination of iron in cells due to the low iron content and the limited growth area of the cells. The method was developed for iron uptake studies for toxicological purposes.

Heavy metals monitoring using bivalves from Mediterranean Sea and Red Sea.

The concentrations of heavy metals (Cd, Co, Cu, Fe, Mn, Ni, Pd and Zn) were measured in the Bivalves (Modiolus auriculatus and Donax trunculus) collected from the Egyptian coasts of Mediterranean Sea and Brachiodonates sp. from the Egyptian coasts of Red Sea. The average concentrations of the heavy metals analyzed exhibited the following decreasing order: Fe > Zn > Cu > Mn > Ni > Co > Pb > Cd for both Mediterranean Sea and Red Sea. The analyses of Cd, Co, Ni, Pb, and Zn showed higher average concentrations for samples collected from Red Sea than that collected from Mediterranean Sea, while Fe, Cu and Mn showed the reverse results. Fe was used as a normalizing agent for all studied metals and exhibited presence of two locations from each of Mediterranean Sea and Red Sea have anthropogenic inputs of heavy metals. These results suggest that the coastal area in both Mediterranean Sea and Red Sea of Egypt might be considered relatively unpolluted with heavy metal.

Speciation of Fe(II) and Fe(III) by the modified ferrozine method, FIA-spectrophotometry, and flame AAS after cloud-point extraction.

A method has been developed for the simultaneous determination of traces of Fe(III) and Fe(II) in water by on-line coupling of spectrophotometry with flame atomic absorption spectrometry (FAAS). The method involves cloud-point extraction (CPE) of both species with ammonium pyrrolidinecarbodithioate (APDC) under standard conditions, which facilitates the in situ complexation and extraction of both species. Differentiation of the oxidation states of iron is achieved by using mathematical equations to overcome the interference of Fe(III) in the spectrophotometric determination of Fe(II) when they are both present in the same solution. In this manner the time-consuming and labor-intensive steps of preoxidation of Fe(II) or reduction of Fe(III) are eliminated. By preconcentrating a 10-mL sample solution detection limits as low as 7 microg L(-1), were obtained after a single-step extraction procedure. The relative standard deviation (n=4, 30 microg L(-1)) was 2.6 % and 1.8 % for spectrophotometry and FAAS, respectively. Recoveries in the range of 96-105 % were obtained by analysis of spiked real samples. The method was further verified by analyzing a certified reference material (IMEP-9); for this the recovery was 98.5 %.

Intravenous versus oral iron supplementation for preoperative stimulation of hemoglobin synthesis using recombinant human erythropoietin.

To compare two modalities of iron supplementation for the preoperative stimulation of erythropoiesis using recombinant human erythropoietin (rhEPO), 12 adults in normal hemoglobin and iron status due for elective surgery were randomized to rhEPO 200 U/kg body weight subcutaneously twice weekly combined with either iron sucrose 200 mg intravenously twice weekly or iron sulfate 160 mg/day orally, for 3 weeks preoperatively. Efficacy was measured by the increases over baseline in hemoglobin, reticulocyte count, and ferritin determined 3 days before surgery; preoperative reticulocyte count and ferritin were significantly higher with intravenous iron, whereas the only significant intragroup increases in hemoglobin between time points also occurred in this group. Intravenous iron significantly boosts the hematopoietic response to rhEPO and prevents iatrogenic iron depletion in otherwise healthy candidates for elective surgery.

Heavy metal contamination of nanosuspensions produced by high-pressure homogenisation.

High pressure homogenisation is a method for the production of nanosuspensions. In this process crystalline drug particles are pressed with high pressure through a narrow homogenisation gap. Due to the conditions in the gap it seems possible that metal erosion can occur. In this study the heavy metal (Fe) contamination of nanosuspensions produced by high pressure homogenisation was determined. Therefore nanosuspensions were analysed by atom absorption spectroscopy concerning their load of iron which is chosen as reference metal. The results show that the erosion of metal is below 1 ppm and will not cause any toxicological problems.

Toxicity considerations when revising the Nordic nutrition recommendations.

The Nordic countries have issued common nutrition recommendations since 1980. In connection with the 3rd revision, a joint working group of nutritionists and toxicologists assessed the toxicology of selected trace elements. Values for upper limits of intake were established for iron, zinc, iodine and selenium. The safety factors between the lowest intakes at which adverse effects had been reported and the suggested upper limits of intake were small. In the toxicological evaluation of upper safe intake levels of essential trace elements, interactions between trace elements as well as long-term exposure to moderately elevated trace element intakes have to be considered.

Serum ferritin iron, a new test, measures human body iron stores unconfounded by inflammation.

Serum ferritin protein is an acute phase reactant. We hypothesized that serum ferritin protein generated in response to an inflammatory process would have much less iron (Fe) in it than would "normal" ferritin protein, and therefore measuring serum ferritin iron would assess human body iron status unconfounded by inflammation. BASIC METHODS: We measured serum ferritin iron in 140 clinical samples obtained from the serum banks of Bronx VA Medical Center Hematology and Nutrition Laboratory (Bronx, NY), the CDC Nutritional Biochemistry serum sample bank (Atlanta, GA), and the sample bank from patients with thalassemia and iron overload treated at New York Hospital (New York, NY). Each was analyzed for three conventional criteria of iron status: serum iron, percentage of transferrin saturation and ferritin protein. In addition, tests for inflammation were also performed: C-reactive protein, WBC and transaminases. Seventy-seven patients' sera from 140 screened met each of three consistent criteria for stages of iron status. Serum ferritin was immobilized by immunoprecipitation with rabbit antihuman polyclonal antibody bound to agarose and separated from other iron-containing proteins, digested with 0.2 ml of 3N nitric acid and analyzed for iron content by atomic absorption spectroscopy. RESULTS: Serum ferritin iron ranged in normal controls from 10 ng to 35 ng Fe/ml. The patients with iron deficiency (4/4) and those in negative iron balance (5/6) had values < or = 10 ng. Positive iron balance (8/9) and iron overload (22/22) values were > 35 ng/ml, in contrast to 11/19 with inflammation. Seventeen of twenty-two with overload had values > 100 ng/ml while only 1/19 with inflammation had such a value. Ferritin iron in ferritin protein was > 15% by weight in 14/22 with iron overload but in 0/19 with inflammation. IMPLICATIONS OF THE WORK: Serum ferritin iron is a simple, direct measure of iron stores that we propose, in conjunction with measuring serum ferritin protein, as a minimally invasive screening procedure for accurately assessing the whole range of human body iron status, unconfounded by inflammation.

Evaluation of multiple micronutrient supplementation in the management of oral submucous fibrosis in Karachi, Pakistan.

Oral submucous fibrosis (OSF) is an oral precancerous condition characterized by symptoms such as intolerance to spicy food, altered salivation, progressive difficulty in opening the mouth, and signs like vesiculation, ulceration, blanching, rigidity, and stiffening of the oral mucosa and depapillation and altered mobility of the tongue. It is seen mostly among people of Indian subcontinent origin. The major structural change is extensive fibroelastic scarring of the lamina propria and deeper connective tissues. A combination of micronutrients (vitamins A, B complex, C, D, and E) and minerals (iron, calcium, copper, zinc, magnesium, and others) was evaluated for its efficacy in controlling the symptoms and signs of OSF in 117 compliant subjects in Karachi, Pakistan, in a single-arm preliminary study. The subjects received supplementation for one to three years. Significant improvement in symptoms, notably intolerance to spicy food, burning sensation, and mouth opening, was observed at exit. The interincisor distance deteriorated in 11 subjects (10%) at exit; it was stable in 56 subjects (49%) and improved in 48 (41%). The mean interincisor distance was 19.1 +/- 10.8 (SD) mm at exit compared with 16.2 +/- 7.5 mm at baseline. A significant proportion of concomitant lesions like leukoplakia also regressed at exit. The major outcome from this study was a beneficial clinical response in subjects with OSF to multiple micronutrient intervention, which justifies its further evaluation in well-designed randomized controlled trials in other settings in South Asia.

Functional and psychic deterioration in elderly people may be aggravated by folate deficiency.

The deterioration of functional and mental capacity is one of the major problems of the elderly. This deterioration may be caused or worsened by folate deficiency. The aim of this investigation was to analyze the relationship between mental and functional capacities and folate status in a group of 177 elderly Spanish people. Folate deficiency is common in the Spanish population. In this study, 48.6% of the elderly subjects had folate intakes below recommended values (200 microg/day), 34.9% had serum concentrations < 14 nmol/L and 6.6% had <360 nmol/L erythrocyte folate. Subjects took part in a series of tests: Katz' scale of activities of daily living, Lawton's scale of instrumental activities of daily living, Pfeiffer's mental status questionnaire, Folstein's Mini-Mental State Test and the Geriatric Depression scale of Yesavage. The results for Lawton's scale of instrumental activities of daily living were significantly better (indicating greater independence and capacity) when folate intake and serum or erythrocyte folate concentrations were adequate (i.e., folate intake no less than recommended, > or = 14 nmol/L serum folate or > or = 360 nmol/L erythrocyte folate). Subjects with adequate Mini-Mental State Exam results (> or = 28 points) had serum and erythrocyte folate concentrations significantly higher than those with less adequate results (<28 points). Thus, there is evidence to suggest that the folate status of the elderly should be monitored and, if possible, improved.

Iron supplementation during pregnancy.

There is world-wide concern that women of childbearing age cannot meet the increased iron needs during pregnancy. In the industrial world however, iron supplementation during pregnancy is a controversial issue. On the one hand, selective iron supplementation after iron status assessment has been recommended because not all women need iron, and compliance is likely to be better when an individual's need is recognized. On the other hand, routine iron supplementation to all women in the second half of pregnancy has been advocated in order to reach all women without the difficulties associated with assessment of iron status. Pregnant women could possibly meet their iron needs by increasing iron absorption efficiency. Although in that case, dietary counselling may prevent anaemia during pregnancy, supplementation with iron will be necessary to cure iron-deficiency anaemia.