Plasma hemoglobin: A method comparison of six assays for hemoglobin and hemolysis index measurement.

INTRODUCTION: Plasma hemoglobin (Hb) is measured for assessment of in vivo and in vitro hemolysis. The objective of the present investigation was to conduct a method comparison of five quantitative and one semi-quantitative Hb and H-index (hemolysis index) assays to evaluate their performance measuring plasma Hb in clinical specimens. METHODS: One hundred and fourteen clinical specimens previously tested for plasma Hb using a laboratory-developed spectrophotometric assay were also tested for Hb using a HemoCue Plasma/Low Hb assay (azide methemoglobin), a laboratory-modified Pointe Scientific Hb assay (cyanmethemoglobin), tested for H-index measurements using a Roche cobas c501, an Abbott Architect c8000, and a semi-quantitative (binned) H-index measurement on a Beckman AU5800. The reference result was defined as the median Hb score (median of all Hb or H-index results). RESULTS: The laboratory-developed spectrophotometric Hb assay and Roche H-index methods mostly closely matched the median Hb score across all data, as well as for lower range median Hb score results ≤2.0 g/L. Two-way frequency table analysis using an Hb (or H-index) cutoff of 0.5 g/L (or 0.5 H-index units) was then performed to compare methods to the median Hb score cutoff. The Beckman method had the highest accuracy at this cutoff, the Roche and Abbott methods had the highest positive predictive value (PPV), and the Beckman, HemoCue, and Pointe methods had the highest negative predictive value (NPV). CONCLUSIONS: Plasma Hb and H-index results vary by method. Laboratories should evaluate the performance characteristics of their respective assays when considering adoption of spectrophotometric or chemical methods for plasma Hb assessment.

Evaluation of microtitre plate-based Haemoglobin estimation.

INTRODUCTION: Haemoglobin estimation is one of the most important clinical investigations. Many techniques are available to measure haemoglobin; still there is a need for a haemoglobin assay technique which is cheap, robust and simple and can be used in field conditions very quickly using figure prick sample. We evaluated a cyanmethaemoglobin-based haemoglobin estimation using a microtitre plates for the purpose. METHODS: Microtitre plate-based haemoglobin estimation was developed using cyanmethaemoglobin-based assay and was compared with standard haematology analyser-based haemoglobin estimation in a large number of samples from a population of voluntary blood donors. Various tests were performed to evaluate the stability of colour, variation of the results during duplicate assay on the same days and on different days as well as linearity of the test was performed against broad range of haemoglobin values for the new microtitre plate-based technique. Standard statistical test of significance was applied to validate the assay. RESULTS: Total 200 samples from in-house and field conditions were evaluated. 10 µL blood sample in 300 µL Drabkin's solution provided optimum and comparable results after 10 minutes of incubation. The colour was stable up to 6 hours, the coefficient of variation was less than 3%, and the cost per test including everything was less than 3 cent/2P. Turnaround time for 90 samples was only 30 minutes. CONCLUSION: Cyanmethaemoglobin-based assay in microtitre plate is feasible, robust, rapid, cheap and cost-effective method for haemoglobin estimation in field conditions.

Determination of the formal redox potentials of the cyanhaemoglobin/cyanmethaemoglobin and the myoglobin/metmyoglobin couples at neutral pH.

Determination of a representative formal redox potential of the Fe(II)/Fe(III) redox couple in cyanhaemoglobin, at pH=7 and related to the state in solution, was the objective of this work. It was achieved at low concentrations of the protein (5µM) to circumvent undesired adsorption. Square-wave voltammetry instead of classical cyclic voltammetry was applied because this method is more sensitive and provides information on the formal redox potential and reversibility, even for rapid processes. We obtained E°'=-0.12±0.01V for cyanhaemoglobin and E°'=-0.10±0.01V, vs. SHE, for myoglobin in comparison. These values differ by only 20mV because the two Fe(II)/Fe(III) redox centres are embedded in closely resembling chemical environments. The small difference is probably owed to the additional axially coordinating cyanide ligand in cyanmethaemoglobin which slightly favours the Fe(III) state in the haem macrocycle.

Updated Method for In Vitro Analysis of Nanoparticle Hemolytic Properties.

Hemolysis is damage to red blood cells (RBCs), which results in the release of the iron-containing protein hemoglobin into plasma. An in vitro assay was developed and described earlier for the analysis of nanoparticle hemolytic properties. Herein, we present a revised version of the original protocol. In this protocol, analyte nanoparticles and controls are incubated in blood. Undamaged RBCs are removed by centrifugation and hemoglobin, released by the damaged erythrocytes, is converted to cyanmethemoglobin by incubation with Drabkin's reagent. The amount of cyanmethemoglobin in the supernatant is measured by spectrophotometry. This measured absorbance is compared to a standard curve to determine the concentration of hemoglobin in the supernatant. The measured hemoglobin concentration is then compared to the total hemoglobin concentration to obtain the percentage of nanoparticle-induced hemolysis. The revision includes updated details about nanoparticle sample preparation, selection of nanoparticle concentration for the in vitro study, updated details about assay controls and case studies about nanoparticle interference with the in vitro hemolysis assay.

[Study on anemia and vitamin A and vitamin D nutritional status of Chinese urban pregnant women in 2010-2012].

Objective: To evaluate the prevalence of anemia and the nutritional status of vitamins A and D by analyzing hemoglobin, serum retinol, and serum 25-hydroxyvitamin D levels in Chinese urban pregnant women during 2010-2012. Methods: Data were obtained from the China Nutrition and Health Survey in 2010-2012. Using multi-stage stratified sampling and population proportional stratified random sampling, 2 250 pregnant women from 34 metropolis and 41 middle-sized and small cities were included in this study. Information was collected using a questionnaire survey. The blood hemoglobin concentration was determined using the cyanmethemoglobin method, and anemia was determined using the World Health Organization guidelines combined with the elevation correction standard. The serum retinol level was determined using high-performance liquid chromatography, and vitamin A deficiency (VAD) was judged by the related standard recommended by the World Health Organization. The vitamin D level was determined using enzyme-linked immunosorbent assay and vitamin D deficiency was judged by the recommendation standards from the Institute of Medicine of The National Academies. The hemoglobin, serum retinol, and serum 25-hydroxyvitamin D levels were compared, along with differences in the prevalence of anemia, VAD, and the vitamin D deficiency rate (including deficiency and serious deficiency). Results: A total of 1 738 cases of hemoglobin level, 594 cases of serum retinol level, and 1 027 cases of serum 25-hydroxyvitamin D were available for analysis in this study. The overall blood hemoglobin level (P(50) (P(25)-P(75))) was 122.70 (114.00-131.10) g/L; 123.70 (115.21-132.00) g/L for metropolis and 122.01 (113.30-130.40) g/L for middle-sized and small cities. The blood hemoglobin level of metropolis residents was significantly higher than that of middle-sized and small city residents (P=0.027). The overall prevalence of anemia was 17.0% (295/1 738). The overall serum retinol level (P(50) (P(25)-P(75))) was 1.61 (1.20-2.06) µmol/L; 1.50 (1.04-2.06) µmol/L for metropolis and 1.63 (1.31-2.05) µmol/L for middle-sized and small cities. The serum retinol level of metropolis residents was significantly higher than that of middle-sized and small city residents (P=0.033). The overall prevalence of VAD was 7.4% (47/639); 11.5% (33/286) for metropolis and 4.0% (14/353) for middle-sized and small cities. A significant difference was observed in the prevalence of VAD between metropolis and middle-sized and small city residents (P<0.001). The overall serum 25-hydroxyvitamin D level (P(50) (P(25)-P(75))) was 15.41 (11.79-20.23) ng/ml; 14.71 (11.15-19.07) ng/ml for metropolis and 16.02 (12.65-21.36) ng/ml for middle-sized and small cities. A significant difference was observed in the vitamin D level between metropolis and middle-sized and small city residents (P<0.001). The overall prevalence of vitamin D deficiency was 74.3% (763/1 027); A significant difference was observed in the prevalence of serious vitamin D deficiency between metropolis (30.64%(144/470)) and middle-sized and small city residents (26%(267/1 027))(P=0.002). There were no significant differences between blood hemoglobin level and the prevalence of anemia, VAD, and vitamin D deficiency. Conclusion: The prevalence of anemia in Chinese urban pregnant women improved from 2002 to 2012. The prevalence of vitamin D deficiency in pregnant women was generally more serious, while a certain percentage of women had VAD. The prevalence of VAD and serious vitamin D deficiency among pregnant women from metropolis was significantly higher than that of pregnant women from medium and small-sized cities.

Visualizing the Bohr effect in hemoglobin: neutron structure of equine cyanomethemoglobin in the R state and comparison with human deoxyhemoglobin in the T state.

Neutron crystallography provides direct visual evidence of the atomic positions of deuterium-exchanged H atoms, enabling the accurate determination of the protonation/deuteration state of hydrated biomolecules. Comparison of two neutron structures of hemoglobins, human deoxyhemoglobin (T state) and equine cyanomethemoglobin (R state), offers a direct observation of histidine residues that are likely to contribute to the Bohr effect. Previous studies have shown that the T-state N-terminal and C-terminal salt bridges appear to have a partial instead of a primary overall contribution. Four conserved histidine residues [αHis72(EF1), αHis103(G10), αHis89(FG1), αHis112(G19) and ßHis97(FG4)] can become protonated/deuterated from the R to the T state, while two histidine residues [αHis20(B1) and ßHis117(G19)] can lose a proton/deuteron. αHis103(G10), located in the α1:ß1 dimer interface, appears to be a Bohr group that undergoes structural changes: in the R state it is singly protonated/deuterated and hydrogen-bonded through a water network to ßAsn108(G10) and in the T state it is doubly protonated/deuterated with the network uncoupled. The very long-term H/D exchange of the amide protons identifies regions that are accessible to exchange as well as regions that are impermeable to exchange. The liganded relaxed state (R state) has comparable levels of exchange (17.1% non-exchanged) compared with the deoxy tense state (T state; 11.8% non-exchanged). Interestingly, the regions of non-exchanged protons shift from the tetramer interfaces in the T-state interface (α1:ß2 and α2:ß1) to the cores of the individual monomers and to the dimer interfaces (α1:ß1 and α2:ß2) in the R state. The comparison of regions of stability in the two states allows a visualization of the conservation of fold energy necessary for ligand binding and release.

[Study on the anemia status of Chinese urban residents in 2010-2012].

OBJECTIVE: The level of blood hemoglobin and the anemia status of Chinese urban residents in 2010-2012 was analyzed. METHODS: All the data in this study came from the China Nutrition and Health Survey in 2010-2012. By using multi-stage stratified sampling and population proportional stratified random sampling method, 74 276 residents aged above 6 from 34 metropolis and 41 middle-sized and small cities were included in this study. The concentration of blood hemoglobin was determined by cyanmethemoglobin method. Anemia was judged by the anemia standard recommended by WHO, combined with elevation correction standard. The level of blood hemoglobin, the prevalence of anemia and the 95%CI value were analyzed by using the complex sampling weighted processing, combined with the population figures released by the National Bureau of Statistics in 2009. RESULTS: In 2010-2012, the level of blood hemoglobin of Chinese city population was(144.16 ± 0.78)g/L, (152.88 ± 0.94)g/L for male and(135.01 ± 0.71)g/L for female, while (145.65 ± 1.22)g/L for metropolis and (143.90 ± 0.89)g/L for small and medium-sized. The anemia prevalence of Chinese city population (pregnant women were not included) was 9.7%(95%CI: 9.4%-10.1%), 6.8%(95%CI: 6.4%-7.3%) for male and 12.8%(95%CI: 12.2%-13.4%) for female, while 8.5%(95% CI: 8.0%-9.0%) for metropolis and 10.0%(95%CI: 9.5%-10.4%) for small and medium-sized. The anemia prevalence of 18-44 women (15.4%, 95%CI: 14.3%-16.6%) was the highest among all the age-groups, and the average anemia prevalence of people more than 60 years-old (including) (12.5%, 95%CI: 11.8%-13.2%) was higher than the other age-groups. CONCLUSION: The anemia prevalence of Chinese city population in 2010-2012 was obviously decreased in comparison of 10 years ago, while, more attention and improvement measures should be take upon women at reproductive age and the elder people.

Comparison of haemoglobin estimates using direct & indirect cyanmethaemoglobin methods.

BACKGROUND & OBJECTIVES: Estimation of haemoglobin is the most widely used method to assess anaemia. Although direct cyanmethaemoglobin method is the recommended method for estimation of haemoglobin, but it may not be feasible under field conditions. Hence, the present study was undertaken to compare indirect cyanmethaemoglobin method against the conventional direct method for haemoglobin estimation. METHODS: Haemoglobin levels were estimated for 888 adolescent girls aged 11-18 yr residing in an urban slum in Delhi by both direct and indirect cyanmethaemoglobin methods, and the results were compared. RESULTS: The mean haemoglobin levels for 888 whole blood samples estimated by direct and indirect cyanmethaemoglobin method were 116.1 ± 12.7 and 110.5 ± 12.5 g/l, respectively, with a mean difference of 5.67 g/l (95% confidence interval: 5.45 to 5.90, P<0.001); which is equivalent to 0.567 g%. The prevalence of anaemia was reported as 59.6 and 78.2 per cent by direct and indirect methods, respectively. Sensitivity and specificity of indirect cyanmethaemoglobin method were 99.2 and 56.4 per cent, respectively. Using regression analysis, prediction equation was developed for indirect haemoglobin values. INTERPRETATION & CONCLUSIONS: The present findings revealed that indirect cyanmethaemoglobin method overestimated the prevalence of anaemia as compared to the direct method. However, if a correction factor is applied, indirect method could be successfully used for estimating true haemoglobin level. More studies should be undertaken to establish agreement and correction factor between direct and indirect cyanmethaemoglobin methods.

Helix-Capping Histidines: Diversity of N-H···N Hydrogen Bond Strength Revealed by (2h)JNN Scalar Couplings.

In addition to its well-known roles as an electrophile and general acid, the side chain of histidine often serves as a hydrogen bond (H-bond) acceptor. These H-bonds provide a convenient pH-dependent switch for local structure and functional motifs. In hundreds of instances, a histidine caps the N-terminus of α- and 310-helices by forming a backbone NH···Nδ1 H-bond. To characterize the resilience and dynamics of the histidine cap, we measured the trans H-bond scalar coupling constant, (2h)JNN, in several forms of Group 1 truncated hemoglobins and cytochrome b5. The set of 19 measured (2h)JNN values were between 4.0 and 5.4 Hz, generally smaller than in nucleic acids (~6-10 Hz) and indicative of longer, weaker bonds in the studied proteins. A positive linear correlation between (2h)JNN and the difference in imidazole ring (15)N chemical shift (Δ(15)N = |δ(15)Nδ1 - δ(15)Nε2|) was found to be consistent with variable H-bond length and variable cap population related to the ionization of histidine in the capping and noncapping states. The relative ease of (2h)JNN detection suggests that this parameter can become part of the standard arsenal for describing histidines in helix caps and other key structural and catalytic elements involving NH···N H-bonds. The combined nucleic acid and protein data extend the utility of (2h)JNN as a sensitive marker of local structural, dynamic, and thermodynamic properties in biomolecules.

An analysis of the bias in red blood cell hemolysis measurement using several analytical approaches.

BACKGROUND: Percentage hemolysis in red cell concentrates (RCC) for transfusion is an indicator of RBC damage. As several factors need to be measured to determine hemolysis, and multiple assays are available for each, the choice of analytical methodology could critically influence results. METHODS: Hemolysis was measured in 48 RCCs every 7 days during storage including expiry (42 days), with supernatant hemoglobin measured using the reference Drabkin's cyanmethemoglobin method and the Harboe spectrophotometric method, total hemoglobin measured using Drabkin's method and 3 automated analyzers (ADVIA 120, CELL-DYN 1700, Coulter AcT), and hematocrit measured using traditional centrifugation and automated analyzers. RESULTS: The choice of method affects hemolysis measurement. Biases ranging from -0.01- -0.03% were observed depending on the combination of methods used. Hematocrit measurement appeared to be a major determinant of bias, and the greatest bias was seen with the ADVIA 120 automated analyzer. Although results did not differ by a level thought to be of clinical significance, the choice of method will impact quality control pass/fail rates. CONCLUSION: Although guidelines exist in many jurisdictions regarding acceptable hemolysis levels in RCCs, these are silent regarding the methods to be used. The presence of bias highlights the need for standardization of methodology.

On the temperature stability of extracellular hemoglobin of Glossoscolex paulistus, at different oxidation states: SAXS and DLS studies.

Glossoscolex paulistus hemoglobin (HbGp) was studied by dynamic light scattering (DLS) and small angle X-ray scattering (SAXS). DLS melting curves were measured for met-HbGp at different concentrations. SAXS temperature studies were performed for oxy-, cyanomet- and met-HbGp forms, at several pH values. At pH 5.0 and 6.0, the scattering curves are identical from 20 to 60 °C, and Rg is 108 Å, independent of the oxidation form. At pH 7.0, protein denaturation and aggregation occurs above 55 °C and 60 °C, for oxy and met-HbGp, respectively. Cyanomet-HbGp, at pH 7.0, is stable up to 60 °C. At alkaline pH (8.0-9.0) and higher temperature, an irreversible dissociation process is observed, with a decrease of Rg, Dmax and I(0). Analysis by p(r), obtained from GNOM, and OLIGOMER, was used to fit the SAXS experimental scattering curves by a combination of theoretical curves obtained for HbLt fragments from the crystal structure. Our results show clearly the increasing contribution of smaller molecular weight fragments, as a function of increasing pH and temperature, as well as, the order of thermal stabilities: cyanomet->oxy->met-HbGp.

Nitric oxide scavenging by red blood cell microparticles and cell-free hemoglobin as a mechanism for the red cell storage lesion.

BACKGROUND: Intravascular red cell hemolysis impairs nitric oxide (NO)-redox homeostasis, producing endothelial dysfunction, platelet activation, and vasculopathy. Red blood cell storage under standard conditions results in reduced integrity of the erythrocyte membrane, with formation of exocytic microvesicles or microparticles and hemolysis, which we hypothesized could impair vascular function and contribute to the putative storage lesion of banked blood. METHODS AND RESULTS: We now find that storage of human red blood cells under standard blood banking conditions results in the accumulation of cell-free and microparticle-encapsulated hemoglobin, which, despite 39 days of storage, remains in the reduced ferrous oxyhemoglobin redox state and stoichiometrically reacts with and scavenges the vasodilator NO. Using stopped-flow spectroscopy and laser-triggered NO release from a caged NO compound, we found that both free hemoglobin and microparticles react with NO about 1000 times faster than with intact erythrocytes. In complementary in vivo studies, we show that hemoglobin, even at concentrations below 10 µmol/L (in heme), produces potent vasoconstriction when infused into the rat circulation, whereas controlled infusions of methemoglobin and cyanomethemoglobin, which do not consume NO, have substantially reduced vasoconstrictor effects. Infusion of the plasma from stored human red blood cell units into the rat circulation produces significant vasoconstriction related to the magnitude of storage-related hemolysis. CONCLUSIONS: The results of these studies suggest new mechanisms for endothelial injury and impaired vascular function associated with the most fundamental of storage lesions, hemolysis.

Method for analysis of nanoparticle hemolytic properties in vitro.

Hemolysis is damage to red blood cells (RBCs), which results in the release of the iron-containing protein hemoglobin into plasma. Here we describe an in vitro assay specifically developed for the analysis of nanoparticle hemolytic properties (see Fig. 1). In this assay, analyte nanoparticles are incubated in blood, and hemoglobin is released by damaged cells and converted to red-colored cyanmethemoglobin by reagents. The nanoparticles and undamaged RBCs are then removed by centrifugation, and the amount of cyanmethemoglobin in the supernatant is measured by spectrophotometry. This measured absorbance is compared to a standard curve to determine the concentration of hemoglobin in the supernatant. This hemoglobin concentration is then compared to that in the supernatant of a blood sample treated with a negative control to obtain the percentage of nanoparticle-induced hemolysis. Fig. 1. Schematic illustration of the steps in this in vitro assay to evaluate nanoparticle hemolytic properties. PFH is plasma-free hemoglobin. CMH is cyanmethemoglobin. TBH is total blood hemoglobin.

Hemoglobin redux: combining neutron and X-ray diffraction with mass spectrometry to analyse the quaternary state of oxidized hemoglobins.

Improvements in neutron diffraction instrumentation are affording the opportunity to re-examine the structures of vertebrate hemoglobins and to interrogate proton and solvent position changes between the different quaternary states of the protein. For hemoglobins of unknown primary sequence, structural studies of cyanomethemoglobin (CNmetHb) are being used to help to resolve sequence ambiguity in the mass spectra. These studies have also provided additional structural evidence for the involvement of oxidized hemoglobin in the process of erythrocyte senescence. X-ray crystal studies of Tibetan snow leopard CNmetHb have shown that this protein crystallizes in the B state, a structure with a more open dyad, which possibly has relevance to RBC band 3 protein binding and erythrocyte senescence. R-state equine CNmetHb crystal studies elaborate the solvent differences in the switch and hinge region compared with a human deoxyhemoglobin T-state neutron structure. Lastly, comparison of histidine protonation between the T and R state should enumerate the Bohr-effect protons.

Preliminary neutron and X-ray crystallographic studies of equine cyanomethemoglobin.

Room-temperature and 100 K X-ray and room-temperature neutron diffraction data have been measured from equine cyanomethemoglobin to 1.7 A resolution using a home source, to 1.6 A resolution on NE-CAT at the Advanced Photon Source and to 2.0 A resolution on the PCS at Los Alamos Neutron Science Center, respectively. The cyanomethemoglobin is in the R state and preliminary room-temperature electron and neutron scattering density maps clearly show the protonation states of potential Bohr groups. Interestingly, a water molecule that is in the vicinity of the heme group and coordinated to the distal histidine appears to be expelled from this site in the low-temperature structure.

International Council for Standardization in Haematology technical report 1-2009: new reference material for haemiglobincyanide for use in standardization of blood haemoglobin measurements.

The International Council for Standardization in Haematology (ICSH) in conjunction with Eurotrol, B.V. has released a new lot of the haemiglobincyanide or haemoglobin standard. This technical report describes the purpose, methodology in manufacturing, summary of value assignment data and availability of this standard material used for the standardization and calibration of whole blood haemoglobin measurements on most haemoglobinometers and automated blood cell counters throughout the world.

Examination of blood haemoglobin concentration measured using the OSM2.

Blood haemoglobin concentration is regularly measured automatically by instruments reporting the value in around 1 min. The OSM2 from Radiometer is an example. Results from this instrument have been compared with those of a reference method using the hemiglobincyanide principle. Four healthy, moderately trained, young men (non-smokers) cycled for 2 min to exhaustion. Blood samples were drawn from indwelling catheters in the femoral artery and vein before exercise, during exercise and in the 1 h recovery. Blood haemoglobin concentration was analysed using both methods. The results of the OSM2 were linearly related to those of the control method, with a random variation of 0.14 mmol L(-1) (1.5%). For arterial blood, the OSM2 showed a systematic bias of -0.36 mmol L(-1) (-4%). For femoral venous samples the bias varied depending on the haemoglobin concentration, being negative at low concentrations and positive at high values (-3 to +2%). Consequently, the arteriovenous (a-v) difference differed systematically between the two methods. The varying bias in the results of the OSM2 for femoral-venous samples correlated with pH, pCO(2), O(2) saturation of haemoglobin (sO(2)) and with the haemoglobin concentration itself (cHb). Partial correlation analyses suggest that only the latter two correlations were independent, while correlations of the bias with pH and pCO(2) were removed when correcting for the effect of sO(2) and cHb. In conclusion, the OSM2 measures the blood haemoglobin concentration fairly precisely, but there is a variable bias of up to 4% in absolute value. Finally, the instrument does not report a-v differences reliably.

Association between magnesium : iron intake ratio and diabetes in Chinese adults in Jiangsu Province.

BACKGROUND: The intake of iron is positively related to the risk of diabetes, whilst magnesium intake is inversely related. However, it is unknown whether there is an interaction between dietary magnesium and iron expressed as a ratio and diabetes. METHODS: This is a cross-sectional household survey carried out in 2002 in Jiangsu Province, China. A total of 2849 men and women aged >or = 20 years participated (participation rate 89.0%). Iron and magnesium intake was assessed by 3-day weighed food records. Fasting plasma glucose, serum ferritin and haemoglobin were measured. RESULTS: The mean intake of iron and magnesium was 25 mg/day and 332 mg/day, respectively. The prevalence of diabetes was 3.0% in men and 2.6% in women. Magnesium intake was negatively associated with diabetes when adjusted for age and sex, but not in a fully adjusted model. A strong inverse association between magnesium : iron intake ratio and diabetes was observed. In the fully adjusted model, the odds ratios of diabetes across quartiles of magnesium : iron intake ratio were: 1.0, 0.63 [95% confidence interval 0.32-1.25], 0.36 (0.16-0.81) and 0.48 (0.20-1.14) (P for trend 0.038). There was an interaction between central obesity and magnesium : iron ratio. CONCLUSION: Magnesium : iron intake ratio is an independent risk marker for diabetes in Chinese adults. As this is a cross-sectional study, we cannot establish any causal relationship.

Zinc supplementation improved length growth only in anemic infants in a multi-country trial of iron and zinc supplementation in South-East Asia.

Data from 4 randomized, placebo-controlled, double-blind trials in Indonesia, Thailand, and Vietnam, the South-East Asian Multicountry Trial on Iron and Zinc supplementation in Infants (SEAMTIZI), were pooled to investigate the effects of iron and zinc supplementation infant growth. Infants (n = 2451) aged 4-6 mo old were supplemented with iron (10 mg/d) and/or zinc (10 mg/d) for 6 mo. Overall, neither iron nor zinc supplementation prevented the progressive growth faltering during infancy, which is common in many developing countries. However, infants who received zinc were less likely to be stunted at the end of the supplementation period (odds ratio 0.80; 95% CI 0.64-1.0). Boys had a 30% higher risk of being stunted at the end of the study than girls (P < 0.01). Baseline factors modified the effect of supplementation, with infants anemic at baseline (hemoglobin < 105 g/L) benefiting from zinc supplementation, with an estimated increase in height-for-age Z-score (HAZ) score of 0.17 (P < 0.01), but with no effect of zinc supplementation on growth in infants not anemic at baseline. Iron supplementation negatively affected linear growth in infants with a birth weight of >3500 g (estimated effect size, -0. 14 HAZ score; P < 0.01), but with no significant effect in infants with a lower birth weight. This study shows that blanket supplementation of infants with iron or zinc will not be beneficial to all recipients and may have adverse effects in some. Hence, interventions such as iron and zinc supplementation for infants should be restricted to subgroups in which there is a clear benefit and baseline factors should be considered and characterized before implementing new policies.

Mechanism of methaemoglobin breakdown by the lysine-specific gingipain of the periodontal pathogen Porphyromonas gingivalis.

Abstract The R- and K-gingipain proteases of Porphyromonas gingivalis are involved in proteolysis of haemoglobin from which the defensive dimeric haem pigment is formed. Whilst oxyhaemoglobin is refractory towards K-gingipain, methaemoglobin is rapidly degraded. Ligation of methaemoglobin with N3-, which effectively blocks haem dissociation from the protein, prevented haemoglobin breakdown. Haem-free globin was rapidly degraded by K-gingipain. These data emphasise the need for haemoglobin oxidation which encourages haem dissociation and makes the haem-free globin susceptible to proteolytic attack.