Diagnostic delays and treatment challenges in children with coeliac disease: The New Zealand Coeliac Health Survey.

AIM: Coeliac disease (CD) is an increasingly common immune-mediated disorder. Treatment is a life-long gluten-free diet. The aim of this study was to describe the presenting symptoms, delays in diagnosis and difficulties associated with managing CD in children. METHOD: The New Zealand Coeliac Health Survey was undertaken in collaboration with Coeliac New Zealand Incorporated, whose membership was the study population. The questionnaire enquired about presenting and ongoing symptoms, and challenges associated with treatment. Children aged <16 years were included in this analysis. Proportions and the mean or median were calculated, as appropriate. RESULTS: There were 123 children with doctor-diagnosed CD. The median age at diagnosis was 4 years (range 0-13 years). The median time between symptom onset and diagnosis was 1.5 years (range 0-11 years). Despite a gluten-free diet, many children continued to experience symptoms, which were most commonly attributed to an unknown cause (61.8%), hidden sources of gluten (44.1%) or food allergy (29.4%). Families found that following a gluten-free diet was very (12%) or moderately (31%) difficult, particularly when eating out. CONCLUSION: Recognition of the challenges associated with the diagnosis and treatment of CD in childhood is an important issue in addressing the needs of children with CD, and their families.

The cost of diabetes-related hospital care to the Southern District Health Board in 2016/17.

AIM: To estimate the cost of diabetes-related hospital admissions to the Southern District Health Board for the year 2016/17. METHODS: Unidentified data with an ICD-10-AM diagnostic code for any type of diabetes were obtained for admissions to Dunedin and Southland Hospitals. Each admission was categorised according to whether the diabetes diagnostic code was listed first, second or subsequently, and by diagnostic group within each of these three categories. The case weight for each admission was multiplied by the 2016/17 cost weight value of NZ$4,824.67. RESULTS: There were 6,994 separate hospital admission events. The total cost was NZ$40,986,618. Admissions where diabetes was the primary, secondary or subsequent diagnosis cost NZ$2,214,172, NZ$8,057,235 and NZ$30,697,210, respectively. More than 80% of admissions were for those aged 55 years and over. Ketoacidosis was the most common primary reason for admission (n=103) among those with type 1 diabetes, costing NZ$349,892. When diabetes was not the primary or secondary diagnosis, the most common primary diagnosis was a circulatory system disease, costing NZ$8,181,324. The mean (SD) cost per admission where the primary diagnosis was coronary artery disease with and without diabetes diagnostic codes was NZ$10,407 ($20,694) and NZ$8,657 ($11,347), respectively. CONCLUSIONS: The annual cost of diabetes-related hospital admissions is substantial. Monitoring the cost of diabetes to DHBs should be prioritised, along with implementation of interventions that reduce preventable diabetes-related hospital admissions, and new diabetes cases.

Addition of Exogenous γ-Glutamyl Hydrolase Eliminates the Need for Lengthy Incubation of Whole-Blood Lysate for Quantitation of Folate Vitamers by High-Performance Liquid Chromatography-Tandem Mass Spectrometry.

BACKGROUND: Measurement of folate monoglutamates by HPLC-tandem mass spectrometry (HPLC-MS/MS) in whole-blood lysate (WBL) requires lengthy incubation before analysis, risking degradation of labile folate vitamers. OBJECTIVE: We explored whether the addition of a commercially available recombinant exogenous γ-glutamyl hydrolase (exoGGH) enzyme reduced the required incubation time of WBL for measurement of folate as monoglutamates. METHODS: For conventional deglutamylation of polyglutamates, WBL was incubated for 4 h at 37°C. Alternatively, we added exoGGH to WBL at varying concentrations (1-10 µg/mL) and incubation times (0-90 min). We also investigated modifications to the sample diluent (pH, ascorbic acid compared with sodium ascorbate, and ascorbate concentration). Finally, we tested the effect of the enzyme in different sample types: WBL from frozen whole blood compared with frozen WBL or with frozen washed RBCs. Samples (n ≤ 15/experiment) were analyzed by HPLC-MS/MS for 6 folate monoglutamates and 5-methyltetrahydrofolate diglutamate. RESULTS: Optimal deconjugation of folate polyglutamates was achieved by using 1% ascorbic acid and 5 µg enzyme/mL WBL, requiring ≤30 min incubation time to achieve complete folate recovery as monoglutamates. This treatment resulted in similar folate concentrations as conventional deglutamylation (4 h at 37°C). The exoGGH enzyme was effective in samples stored frozen as whole blood and as WBL. However, the extended thaw time of whole blood resulted in 5-methyltetrahydrofolate loss and unacceptable changes to the non-methyl folate concentration. Total folate (with exoGGH) measured in washed RBCs was ∼15% lower than RBC folate calculated from WBL concentrations (conventional deglutamylation). CONCLUSIONS: The use of exoGGH minimized incubation time and thus may avoid degradative losses of labile folate forms during sample preparation. The lower folate results in washed RBCs may be due to inadequate packing of RBCs, among other unidentified factors. A larger study is required to confirm the lack of differences in folate concentrations determined with and without the use of exoGGH.

Lactating Canadian Women Consuming 1000 µg Folic Acid Daily Have High Circulating Serum Folic Acid Above a Threshold Concentration of Serum Total Folate.

Background: Consumption of high-dose folic acid supplements is common throughout pregnancy and lactation in several countries, including Canada, Brazil, and the United States, and may lead to high levels of circulating unmetabolized folic acid. Objective: The objective of the study was to characterize serum and whole-blood folate forms in Canadian lactating women regularly consuming a daily high-dose folic acid supplement. Methods: One-hundred and seventeen Canadian lactating women aged between 18 and 42 y, with a geometric mean ± SD prepregnancy body mass index (kg/m2) of 23.1 ± 1.2, were enrolled in a vitamin D supplementation trial between 13 and 22 wk of gestation. As part of the trial, the women received a daily multivitamin containing 1000 µg folic acid throughout pregnancy and lactation until 8 wk postpartum. At 8 wk postpartum, serum folate forms, including folic acid and RBC total folate, were determined from nonfasted blood samples. Differences in median folate vitamer concentrations among quintiles of serum total folate status were assessed by the Wald test and quantile regression methods. A breakpoint in the relation between serum folic acid and serum total folate was modeled with the use of the segmented package in R. Results: Median serum total folate concentration among participants was 79.3 nmol/L (5th-95th percentile 30.7-186 nmol/L) and median RBC folate concentration was 2790 nmol/L (5th-95th percentile 1330-4850 nmol/L). There was a breakpoint in the relation between serum total folate and serum folic acid at 78.5 nmol/L (95% CI: 67.9, 89.1 nmol/L), below which serum folic acid was not associated with serum total folate, and above which serum folic acid increased 0.78 nmol/L (95% CI: 0.70, 0.86 nmol/L; P < 0.001) for each 1 nmol/L increase in serum total folate. Conclusions: These data demonstrate the potential for high serum folic acid concentrations proportional to overall folate concentrations in lactating women with serum total folate >80 nmol/L taking high-dose supplemental folic acid. This study was registered at clinicaltrials.gov as NCT01112891.

Iron, Zinc, Folate, and Vitamin B-12 Status Increased among Women and Children in Yaoundé and Douala, Cameroon, 1 Year after Introducing Fortified Wheat Flour.

Background: Few data are available on the effectiveness of large-scale food fortification programs.Objective: We assessed the impact of mandatory wheat flour fortification on micronutrient status in Yaoundé and Douala, Cameroon.Methods: We conducted representative surveys 2 y before and 1 y after the introduction of fortified wheat flour. In each survey, 10 households were selected within each of the same 30 clusters (n = ∼300 households). Indicators of inflammation, malaria, anemia, and micronutrient status [plasma ferritin, soluble transferrin receptor (sTfR), zinc, folate, and vitamin B-12] were assessed among women aged 15-49 y and children 12-59 mo of age.Results: Wheat flour was consumed in the past 7 d by ≥90% of participants. Postfortification, mean total iron and zinc concentrations of flour samples were 46.2 and 73.6 mg/kg (target added amounts were 60 and 95 mg/kg, respectively). Maternal anemia prevalence was significantly lower postfortification (46.7% compared with 39.1%; adjusted P = 0.01), but mean hemoglobin concentrations and child anemia prevalence did not differ. For both women and children postfortification, mean plasma concentrations were greater for ferritin and lower for sTfR after adjustments for potential confounders. Mean plasma zinc concentrations were greater postfortification and the prevalence of low plasma zinc concentration in women after fortification (21%) was lower than before fortification (39%, P < 0.001); likewise in children, the prevalence postfortification (28%) was lower than prefortification (47%, P < 0.001). Mean plasma total folate concentrations were ∼250% greater postfortification among women (47 compared with 15 nmol/L) and children (56 compared with 20 nmol/L), and the prevalence of low plasma folate values was <1% after fortification in both population subgroups. In a nonrepresentative subset of plasma samples, folic acid was detected in 77% of women (73% of those fasting) and 93% of children. Mean plasma and breast-milk vitamin B-12 concentrations were >50% greater postfortification.Conclusion: Although the pre-post survey design limits causal inference, iron, zinc, folate, and vitamin B-12 status increased among women and children in urban Cameroon after mandatory wheat flour fortification.

Correlations between Maternal, Breast Milk, and Infant Vitamin B12 Concentrations among Mother-Infant Dyads in Vancouver, Canada and Prey Veng, Cambodia: An Exploratory Analysis.

Vitamin B12 plays an essential role in fetal and infant development. In regions where animal source food consumption is low and perinatal supplementation is uncommon, infants are at risk of vitamin B12 deficiency. In this secondary analysis, we measured total vitamin B12 concentrations in maternal and infant serum/plasma and breast milk among two samples of mother-infant dyads in Canada (assessed at 8 weeks post-partum) and in Cambodia (assessed between 3-27 weeks post-partum). Canadian mothers (n = 124) consumed a daily vitamin B12-containing multiple micronutrient supplement throughout pregnancy and lactation; Cambodian mothers (n = 69) were unsupplemented. The maternal, milk, and infant total vitamin B12 concentrations (as geometric means (95% CI) in pmol/L) were as follows: in Canada, 698 (648,747), 452 (400, 504), and 506 (459, 552); in Cambodia, 620 (552, 687), 317 (256, 378), and 357 (312, 402). The majority of participants were vitamin B12 sufficient (serum/plasma total B12 > 221 pmol/L): 99% and 97% of mothers and 94% and 84% of infants in Canada and Cambodia, respectively. Among the Canadians, maternal, milk, and infant vitamin B12 were all correlated (p < 0.05); only maternal and infant vitamin B12 were correlated among the Cambodians (p < 0.001).

Nutrient intake values for folate during pregnancy and lactation vary widely around the world.

Folate is a B-vitamin with particular importance during reproduction due to its role in the synthesis and maintenance of DNA. Folate is well known for its role in preventing neural tube defects (NTDs) during the periconceptional period. There is also an increased need for folate throughout pregnancy to support optimal growth and development of the fetus and blood volume expansion and tissue growth of the mother. During lactation, women are at risk of folate deficiency due to increased demands to accommodate milk folate levels. Nutrient Intake Values (NIVs) for folate have been calculated to take into account additional needs during pregnancy and lactation. However, these values vary widely between countries. For example, the folate requirement that is set to meet the needs of almost all healthy women during pregnancy varies from 300 µg/day in the United Kingdom to 750 µg/day in Mexico. Currently, there is no accepted standardized terminology or framework for establishing NIVs. This article reviews country-specific NIVs for folate during pregnancy and lactation and the basis for setting these reference values.

Quantitation of whole-blood total folate within defined MTHFR C677T genotype groups by isotope dilution-liquid chromatography-tandem mass spectrometry differs from microbiologic assay.

Standardization of folate measurement is needed for accurate assessment of folate status. We compared the measurement of whole-blood folate by isotope dilution-liquid chromatography-tandem MS (ID-LC-MS/MS) with the historical gold standard microbiological assay (MA) using 3 common calibrators within the frame of the methylenetetrahydrofolate reductase (MTHFR) C677T polymorphism. Seventy-three whole-blood samples with an even distribution of MTHFR C677T genotypes (24 CC, 24 CT, 24 TT) were prepared, and total folate was determined by ID-LC-MS/MS and MA using the following calibrators: 5-methyltetrahydrofolate (5-methylTHF) (Merck), folic acid (FA) (Merck), and FA (Sigma). To compare the methods, 5-formyltetrahydrofolate (5-formylTHF) was excluded in the ID-LC-MS/MS summation of total folate, because it is likely that the majority of 5-formylTHF detected is a pyrazino-s-triazine oxidation product of 5-methylTHF. MA whole-blood folate measured by using the FA calibrators was consistently higher than with the 5-methylTHF calibrator. Differences between dilutions and analysis of spiked whole-blood samples showed a nonlinear response, with overrecovery of 5-methylTHF by ~23% toward the higher end of the MA calibration range. Significant proportional biases between ID-LC-MS/MS and MA were found in all comparisons except when the MA was calibrated with 5-methylTHF and a higher sample dilution of 1:1600 (regression slope: 1.05; P = 0.31; intercept-21, P = 0.16). Calibration bias and matrix effects in the MA underscore the need for a formally accepted whole-blood folate reference method. ID-LC-MS/MS procedures have the potential to offer a high degree of accuracy; however, further work is needed to determine the origin of the pyrazino-s-triazine derivative.

A hydrogen peroxide electrode assay to measure thiol peroxidase activity for organoselenium and organotellurium drugs.

Molecular mimics of the enzyme glutathione peroxidase (GPx) are increasingly being evaluated as redox active drugs. Their molecular mechanism of action parallels that of the native enzyme; however, a major distinction is that GPx mimics can use alternative thiol substrates to glutathione. This generic thiol peroxidase activity implies that it is necessary to assess a GPx mimic's recognition of a range of cellular thiols in order to determine its potential therapeutic effects. We report an electrochemical assay that, by measuring the rate of decrease of the peroxide substrate, allows the activity of GPx mimics to be directly compared against an array of thiols. The derived pseudo zero-order rate constants, k(obs), for representative GPx mimics range between 0 and 6.6 min(-1) and can vary by more than an order of magnitude depending on the thiol electron donor. An additional advantage of the assay is that it enables synergistic interactions between GPx mimics and cellular proteins to be evaluated. Here we report that glutathione disulfide reductase, which is commonly used to evaluate GPx mimic activity, recognizes the GPx mimic ebselen as a substrate, increasing its apparent k(obs). Therefore, reports relying on glutathione disulfide reductase to evaluate GPx mimic activity may exaggerate drug antioxidant action.