Iron supplements acutely increase hepcidin, but the duration and magnitude of the increase, its dose dependence, and its effects on subsequent iron absorption have not been characterized in humans. ...Better understanding of these phenomena might improve oral iron dosing schedules. We investigated whether the acute iron-induced increase in hepcidin influences iron absorption of successive daily iron doses and twice-daily iron doses. We recruited 54 nonanemic young women with plasma ferritin ≤20 µg/L and conducted: (1) a dose-finding investigation with 40-, 60-, 80-, 160-, and 240-mg labeled Fe as 57Fe-, 58Fe-, or 54Fe-FeSO4 given at 8:00 am fasting on 1 or on 2 consecutive days (study 1, n = 25; study 2, n = 16); and (2) a study giving three 60-mg Fe doses (twice-daily dosing) within 24 hours (study 3, n = 13). In studies 1 and 2, 24 hours after doses ≥60 mg, serum hepcidin was increased (P < .01) and fractional iron absorption was decreased by 35% to 45% (P < .01). With increasing dose, fractional absorption decreased (P < .001), whereas absolute absorption increased (P < .001). A sixfold increase in iron dose (40-240 mg) resulted in only a threefold increase in iron absorbed (6.7-18.1 mg). In study 3, total iron absorbed from 3 doses (2 mornings and an afternoon) was not significantly greater than that from 2 morning doses. Providing lower dosages (40-80 mg Fe) and avoiding twice-daily dosing maximize fractional absorption. The duration of the hepcidin response supports alternate day supplementation, but longer-term effects of these schedules require further investigation. These clinical trials were registered at www.ClinicalTrials.gov as #NCT01785407 and #NCT02050932.
•Iron supplements at doses of 60 mg Fe as FeSO4 or higher increase hepcidin for up to 24 hours and are associated with lower iron absorption on the following day.•The soluble transferrin receptor/ferritin ratio and hepcidin are equivalent predictors of iron absorption from supplements.
In iron-depleted women without anemia, oral iron supplements induce an increase in serum hepcidin (SHep) that persists for 24 hours, decreasing iron absorption from supplements given later on the ...same or next day. Consequently, iron absorption from supplements is highest if iron is given on alternate days. Whether this dosing schedule is also beneficial in women with iron-deficiency anemia (IDA) given high-dose iron supplements is uncertain. The primary objective of this study was to assess whether, in women with IDA, alternate-day administration of 100 and 200 mg iron increases iron absorption compared to consecutive-day iron administration. Secondary objectives were to correlate iron absorption with SHep and iron status parameters. We performed a cross-over iron absorption study in women with IDA (n=19; median hemoglobin 11.5 mg/dL; mean serum ferritin 10 mg/L) who received either 100 or 200 mg iron as ferrous sulfate given at 8 AM on days 2, 3 and 5 labeled with stable iron isotopes 57Fe, 58Fe and 54Fe; after a 16-day incorporation period, the other labeled dose was given at 8 AM on days 23, 24 and 26 (days 2, 3 and 5 of the second period). Iron absorption on days 2 and 3 (consecutive) and day 5 (alternate) was assessed by measuring erythrocyte isotope incorporation. For both doses, SHep was higher on day 3 than on day 2 (
<0.001) or day 5 (
<0.01) with no significant difference between days 2 and 5. Similarly, for both doses, fractional iron absorption (FIA) on days 2 and 5 was 40-50% higher than on day 3 (
<0.001), while absorption on day 2 did not differ significantly from day 5. There was no significant difference in the incidence of gastrointestinal side effects comparing the two iron doses (
=0.105). Alternate day dosing of oral iron supplements in anemic women may be preferable because it sharply increases FIA. If needed, to provide the same total amount of iron with alternate day dosing, twice the daily target dose should be given on alternate days, as total iron absorption from a single dose of 200 mg given on alternate days was approximately twice that from 100 mg given on consecutive days (
<0.001). In IDA, even if hepatic hepcidin expression is strongly suppressed by iron deficiency and erythropoietic drive, the intake of oral iron supplements leads to an acute hepcidin increase for 24 hours. The study was funded by ETH Zürich, Switzerland. This study has been registered at
.
Bifidobacteria is one of the major gut commensal groups found in infants. Their colonization is commonly associated with beneficial effects to the host through mechanisms like niche occupation and ...nutrient competition against pathogenic bacteria. Iron is an essential element necessary for most microorganisms, including bifidobacteria and efficient competition for this micronutrient is linked to proliferation and persistence. For this research we hypothesized that bifidobacteria in the gut of iron deficient infants can efficiently sequester iron. The aim of the present study was to isolate bifidobacteria in fecal samples of iron deficient Kenyan infants and to characterize siderophore production and iron internalization capacity.
Fifty-six bifidobacterial strains were isolated by streaking twenty-eight stool samples from Kenyan infants, in enrichment media. To target strains with high iron sequestration mechanisms, a strong iron chelator 2,2-dipyridyl was supplemented to the agar media. Bifidobacterial isolates were first identified to species level by 16S rRNA sequencing, yielding B. bifidum (19 isolates), B. longum (15), B. breve (11), B. kashiwanohense (7), B. pseudolongum (3) and B. pseudocatenulatum (1). While most isolated bifidobacterial species are commonly encountered in the infantile gut, B. kashiwanohense was not frequently reported in infant feces. Thirty strains from culture collections and 56 isolates were characterized for their siderophore production, tested by the CAS assay. Siderophore activity ranged from 3 to 89% siderophore units, with 35 strains (41%) exhibiting high siderophore activity, and 31 (36%) and 20 (23%) showing intermediate or low activity. The amount of internalized iron of 60 bifidobacteria strains selected for their siderophore activity, was in a broad range from 8 to118 μM Fe. Four strains, B. pseudolongum PV8-2, B. kashiwanohense PV20-2, B. bifidum PV28-2a and B. longum PV5-1 isolated from infant stool samples were selected for both high siderophore activity and iron internalization.
A broad diversity of bifidobacteria were isolated in infant stools using iron limited conditions, with some strains exhibiting high iron sequestration properties. The ability of bifidobacteria to efficiently utilize iron sequestration mechanism such as siderophore production and iron internalization may confer an ecological advantage and be the basis for enhanced competition against enteropathogens.
Hepatic hepcidin synthesis is stimulated by inflammation but inhibited during iron deficiency anemia (IDA). In humans, the relative strength of these opposing signals on serum hepcidin and the net ...effect on iron absorption and systemic iron recycling is uncertain. In this prospective, 45-day study, in young women (n=46; age 18-49 years) with or without IDA, we compared iron and inflammation markers, serum hepcidin and erythrocyte iron incorporation from
Fe-labeled test meals, before and 8, 24 and 36 hours (h) after influenza/DPT vaccination as an acute inflammatory stimulus. Compared to baseline, at 24-36 h after vaccination: 1) interleukin-6 increased 2-3-fold in both groups (
<0.001); 2) serum hepcidin increased >2-fold in the non-anemic group (
<0.001), but did not significantly change in the IDA group; 3) serum iron decreased in the non-anemic group (
<0.05) but did not change in the IDA group; and 4) erythrocyte iron incorporation did not change in either of the two groups, but was approximately 2-fold higher in the IDA group both before and after vaccination (
<0.001). In this study, mild acute inflammation did not increase serum hepcidin in women with IDA, suggesting low iron status and erythropoietic drive offset the inflammatory stimulus on hepcidin expression. In non-anemic women, inflammation increased serum hepcidin and produced mild hypoferremia, but did not reduce dietary iron absorption, suggesting iron-recycling macrophages are more sensitive than the enterocyte to high serum hepcidin during inflammation. The study was registered as a prospective observational trial at
The study was funded by the International Atomic Energy Agency.
Sweetpotato and potato are fast-maturing staple crops and widely consumed in low- and middle-income countries. Conventional breeding to biofortify these crops with iron could improve iron intakes. To ...our knowledge, iron absorption from sweetpotato and potato has not been assessed.
The aim was to assess iron absorption from regular and iron-biofortified orange-fleshed sweetpotato in Malawi and yellow-fleshed potato and iron-biofortified purple-fleshed potato in Peru.
We conducted 2 randomized, multiple-meal studies in generally healthy, iron-depleted women of reproductive age. Malawian women (n = 24) received 400 g regular or biofortified sweetpotato test meals and Peruvian women (n = 35) received 500 g regular or biofortified potato test meals. Women consumed the meals at breakfast for 2 wk and were then crossed over to the other variety. We labeled the test meals with 57Fe or 58Fe and measured cumulative erythrocyte incorporation of the labels 14 d after completion of each test-meal sequence to calculate iron absorption. Iron absorption was compared by paired-sample t tests.
The regular and biofortified orange-fleshed sweetpotato test meals contained 0.55 and 0.97 mg Fe/100 g. Geometric mean (95% CI) fractional iron absorption (FIA) was 5.82% (3.79%, 8.95%) and 6.02% (4.51%, 8.05%), respectively (P = 0.81), resulting in 1.9-fold higher total iron absorption (TIA) from biofortified sweetpotato (P < 0.001). The regular and biofortified potato test meals contained 0.33 and 0.69 mg Fe/100 g. FIA was 28.4% (23.5%, 34.2%) from the regular yellow-fleshed and 13.3% (10.6%, 16.6%) from the biofortified purple-fleshed potato meals, respectively (P < 0.001), resulting in no significant difference in TIA (P = 0.88).
FIA from regular yellow-fleshed potato was remarkably high, at 28%. Iron absorbed from both potato test meals covered 33% of the daily absorbed iron requirement for women of reproductive age, while the biofortified orange-fleshed sweetpotato test meal covered 18% of this requirement. High polyphenol concentrations were likely the major inhibitors of iron absorption. These trials were registered at www.clinicaltrials.gov as NCT03840031 (Malawi) and NCT04216030 (Peru).
Plasma ferritin is a widely used indicator to detect iron deficiency, but the threshold ferritin that defines iron deficiency remains uncertain. Our aim was to define the ferritin concentration at ...which the body begins to upregulate iron absorption from the diet; this could provide a functionally-defined threshold of incipient iron deficiency. We hypothesized this threshold ferritin concentration would correspond to the threshold hepcidin concentration at which iron absorption begins to increase.
We performed a pooled analysis of our stable iron isotope studies (n = 1058) conducted from 2006 to 2019 in healthy women (age 18–50 years; mean±SD ferritin 33.7 ± 27.1 μg/L) that measured iron absorption from labeled test meals providing physiological amounts of iron. To fit relationships between iron absorption, ferritin and hepcidin, we used generalized additive modeling, and to identify thresholds, we estimated the first derivatives of the fitted trend to assess inflection points in these relationships.
Hepcidin increased linearly with increasing ferritin over the entire range of ferritin values. Iron absorption began to increase below a threshold hepcidin value of 3.09 (95%CI: 2.80, 3.38) nmol/l, above which iron absorption remained stable. Iron absorption began to increase below a threshold ferritin value of 51.1 (95%CI: 49.1, 53.1) µg/l, above which iron absorption remained stable. The latter two findings were internally consistent in that, in the relationship between hepcidin and ferritin, a hepcidin of ~3 nmol/l corresponded to a ferritin of ~51 µg/l.
Based on physiological upregulation of iron absorption, a threshold ferritin of <50 µg/L, corresponding to a threshold hepcidin of <3 nmol/l, indicates incipient iron deficiency in young women.
Funding for this study was provided by the ETH Zurich.
Genome wide studies have associated TMPRSS6 rs855791 (2321 C>T) with iron status
and hepcidin. It is unclear whether this polymorphism affects iron absorption. In
nonanemic Taiwanese women (n=79, 44 ...TT variant, 35 CC variant), we administered
standardized rice-based test meals containing 4 mg of labeled 57Fe or 58Fe as FeSO4 on
alternate days. Fractional iron absorption was measured by erythrocyte incorporation of
the tracers 14 days after administration. Compared to the CC variant, in the TT variant
serum iron and transferrin saturation were lower (P=0.001; P<0.001, respectively) and
serum hepcidin/transferrin saturation and serum hepcidin/serum iron ratios were higher
(P=0.042; P=0.088, respectively). Serum hepcidin did not differ between groups
(P=0.862). Geometric mean (95% CI) fractional iron absorption, corrected to a serum
ferritin of 15 μg/L, was 26.6% (24.0, 29.5) in the CC variant and 18.5% (16.2, 21.1) in the
TT variant (P=0.002). Overall, predictors of iron absorption were: serum ferritin
(P<0.001); genetic variant (P=0.032); and hepcidin (P<0.001). In the models by variant,
in the CC variant the model explained 67-71% of variability in absorption and serum
ferritin was the only significant predictor (P<0.001); in the TT variant, the model
explained only 35-43% of variability, and hemoglobin (P=0.032), soluble transferrin
receptor (P=0.004) and hepcidin (P<0.001) were significant predictors. Women with the
TMPRSS6 rs855791 (2321 C>T) polymorphism show altered iron homeostasis which
affects oral iron absorption and may increase their risk for iron deficiency. The trial was
registered at www.clinicaltrials.gov as NCT03317873, and funded by the Kaohsiung
Chang-Gung Memorial Hospital, Kaohsiung, Taiwan, (grant CMRPG8F0721) and ETH
Zurich, Switzerland.
Bouillon cubes are widely consumed and when fortified with iron could contribute in preventing iron deficiency. We report the development (part I) and evaluation (current part II) of a novel ferric ...phytate compound to be used as iron fortificant in condiments such as bouillon. Ferric pyrophosphate (FePP), is the compound of choice due to its high stability in foods, but has a modest absorption in humans. Our objective was to assess iron bioavailability from a novel iron fortificant consisting of ferric iron complexed with phytic acid and hydrolyzed corn protein (Fe-PA-HCP), used in bouillon with and without an inhibitory food matrix. In a randomised single blind, cross-over study, we measured iron absorption in healthy adult women (n = 22). In vitro iron bioaccessibility was assessed using a Caco-2 cell model. Iron absorption from Fe-PA-HCP was 1.5% and 4.1% in bouillon with and without inhibitory matrix, respectively. Relative iron bioavailability to FeSO
was 2.4 times higher than from FePP in bouillon (17% vs 7%) and 5.2 times higher when consumed with the inhibitory meal (41% vs 8%). Similar results were found in vitro. Fe-PA-HCP has a higher relative bioavailability versus FePP, especially when bouillon is served with an inhibitory food matrix.
Direct iodization in fish sauce, soy sauce, and seasoning sauces plays a crucial role in optimizing the iodine intake of Thailand’s people. However, determining the iodine content to ensure that ...these sauces meet the standard of Thailand’s Food and Drug Administration (FDA) is challenging. In this study, all local laboratories equipped with inductively coupled plasma–mass spectrometry (ICP-MS) and with experience in iodine analysis by any analytical method were invited to participate in a hands-on training workshop and two rounds of interlaboratory comparison. The aim was to improve laboratory performance and assess the potential for iodine monitoring for mandatory direct-iodized sauces. All target laboratories participated in this study. The hands-on training workshop harmonized the analytical method and increased the capacity of participating laboratories. Most laboratories (7/8) achieved satisfactory performance for six test samples based on interlaboratory comparison. Samples were extracted by tetramethylammonium hydroxide (TMAH), with the presence of 6% 2-propanol, 0.01% triton X-100, internal standard, and iodine determination in direct-iodized sauces by ICP-MS. The reproducibility standard deviation (SL), after the removal of outlier results for iodine content, was 7–22% iodine at a level of 0.03–4.81 mg/L. Moreover, the Thai FDA’s judgment range for official control activities should expand the range of 2–3 mg per 1 L (ppm) by at least 22%.
Organic fertilizer applications can contribute to Zinc (Zn) biofortification of crops. An enriched stable isotope source tracing approach is a central tool to further determine the potential of this ...biofortification measure. Here, we assessed the use of the widely available quadrupole single-collector ICPMS (Q-ICPMS, analytical error = 1% relative standard deviation) and the less accessible but more precise multicollector ICPMS as reference instrument (MC-ICPMS, analytical error = 0.01% relative standard deviation) to measure enriched Zn stable isotope ratios in soil-fertilizer-plant systems. The isotope label was either applied to the fertilizer (direct method) or to the soil available Zn pool that was determined by isotope ratios measurements of the shoots that grew on labeled soils without fertilizer addition (indirect method). The latter approach is used to trace Zn that was added to soils with complex insoluble organic fertilizers that are difficult to label homogeneously. To reduce isobaric interferences during Zn isotope measurements, ion exchange chromatography was used to separate the Zn from the sample matrix. The
Zn:
Zn isotope ratios altered from 0.148 at natural abundance to 1.561 in the fertilizer of the direct method and 0.218 to 0.305 in soil available Zn of the indirect method. Analysis of the difference (Bland-Altman) between the two analytical instruments revealed that the variation between
Zn:
Zn isotope ratios measured with Q-ICPMS and MC-ICPMS were on average 0.08% 95% confidence interval (CI) = 0.68%. The fractions of Zn derived from the fertilizer in the plant were on average 0.16% higher (CI = 0.49%) when analyzed with Q- compared to MC-ICPMS. The sample matrix had a larger impact on isotope measurements than the choice of analytical instrument, as non-purified samples resulted on average 5.79% (CI = 9.47%) higher isotope ratios than purified samples. Furthermore, the gain in analytical precision using MC-ICPMS instead of Q-ICPMS was small compared to the experimental precision. Thus, Zn isotope measurements of purified samples measured with Q-ICPMS is a valid method to trace Zn sources in soil-fertilizer-plant systems. For the indirect source tracing approach, we outlined strategies to sufficiently enrich the soil with Zn isotopes without significantly altering the soil available Zn pool.
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