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Background: In the past, elevated estradiol levels were reported to downregulate the iron regulatory hormone hepcidin, thereby potentially improving iron metabolism. As estrogen plays a role in regulating the menstrual cycle and can influence the cytokine interleukin-6 (IL-6; a hepcidin up-regulator), this investigation examined the effects of estradiol supplementation achieved by the use of a monophasic oral contraceptive pill (OCP) on IL-6, hepcidin levels and iron status during the hormone-deplete versus hormone-replete phases within an oral contraceptive cycle (OCC). Methods: Fifteen healthy female OCP users were recruited and provided a venous blood sample on 2 separate mornings during a 28-day period. These included (a) days 2-4 of the OCC, representing a hormone-free withdrawal period (WD); (b) days 12-14 of the OCC, representing the end of the first week of active hormone therapy (AHT). Results: IL-6 and hepcidin levels were not significantly different at WD and AHT. Serum ferritin was significantly higher (p = 0.039) during AHT as compared to WD. Conclusions: Fluctuations in OCP hormones (estradiol and/or progestogen) had no effect on basal IL-6 and hepcidin levels in young women. Nevertheless, elevated ferritin levels recorded during AHT may indicate that OCP hormones can positively influence iron stores within an OCC despite unchanged hepcidin levels.

In physiological conditions, circulating iron can be filtered by the glomerulus and is almost completely reabsorbed by the tubular epithelium to prevent urinary iron wasting. Increased urinary iron concentrations have been associated with renal injury. However, it is not clear whether increased urinary iron concentrations in patients are the result of increased glomerular iron filtration and/or insufficient tubular iron reabsorption and if these processes contribute to renal injury. We measured plasma and urine iron parameters and urinary tubular injury markers in healthy human subjects ( n = 20), patients with systemic iron overload ( n = 20), and patients with renal tubular dysfunction ( n = 18). Urinary iron excretion parameters were increased in both patients with systemic iron overload and tubular dysfunction, whereas plasma iron parameters were only increased in patients with systemic iron overload. In patients with systemic iron overload, increased urinary iron levels were associated with elevated circulating iron, as indicated by transferrin saturation (TSAT), and increased body iron, as suggested by plasma ferritin concentrations. In patients with tubular dysfunction, enhanced urinary iron and transferrin excretion were associated with distal tubular injury as indicated by increased urinary glutathione S-transferase pi 1-1 (GSTP1-1) excretion. In systemic iron overload, elevated urinary iron and transferrin levels were associated with increased injury to proximal tubules, indicated by increased urinary kidney injury marker 1 (KIM-1) excretion. Our explorative study demonstrates that both glomerular filtration of elevated plasma iron levels and insufficient tubular iron reabsorption could increase urinary iron excretion and cause renal injury.

Orally administrated iron is suspected to increase susceptibility to enteric infections among children in infection endemic regions. Here we investigated the effect of dietary iron on the pathology and local immune responses in intestinal infection models. Mice were held on iron-deficient, normal iron, or high iron diets and after 2 weeks they were orally challenged with the pathogen Citrobacter rodentium. Microbiome analysis by pyrosequencing revealed profound iron- and infection-induced shifts in microbiota composition. Fecal levels of the innate defensive molecules and markers of inflammation lipocalin-2 and calprotectin were not influenced by dietary iron intervention alone, but were markedly lower in mice on the iron-deficient diet after infection. Next, mice on the iron-deficient diet tended to gain more weight and to have a lower grade of colon pathology. Furthermore, survival of the nematode Caenorhabditis elegans infected with Salmonella enterica serovar Typhimurium was prolonged after iron deprivation. Together, these data show that iron limitation restricts disease pathology upon bacterial infection. However, our data also showed decreased intestinal inflammatory responses of mice fed on high iron diets. Thus additionally, our study indicates that the effects of iron on processes at the intestinal host-pathogen interface may highly depend on host iron status, immune status, and gut microbiota composition.

Item does not contain fulltext BACKGROUND: The development of atherosclerosis may be enhanced by iron accumulation in macrophages. Hepcidin-25 is a key regulator of iron homeostasis, which downregulates the cellular iron exporter ferroportin. In haemodialysis (HD) patients, hepcidin-25 levels are increased. Therefore, it is conceivable that hepcidin-25 is associated with all-cause mortality and/or fatal and non-fatal cardiovascular (CV) events in this patient group. The aim of the current analysis was to study the relationship between hepcidin-25 and all-cause mortality and both fatal and non-fatal CV events in chronic HD patients. METHODS: Data from 405 chronic HD patients included in the CONvective TRAnsport STudy (NCT00205556) were studied (62% men, age 63.7 +/- 13.9 years [mean +/- SD]). The median (range) follow-up was 3.0 (0.8-6.6) years. Hepcidin-25 was measured with mass spectrometry. The relationship between hepcidin-25 and all-cause mortality or fatal and non-fatal CV events was investigated with multivariate Cox proportional hazard models. RESULTS: Median (interquartile range) hepcidin-25 level was 13.8 (6.6-22.5) nmol/L. During follow-up, 158 (39%) patients died from any cause and 131 (32%) had a CV event. Hepcidin-25 was associated with all-cause mortality in an unadjusted model [hazard ratio (HR) 1.14 per 10 nmol/L, 95% CI 1.03-1.26; P = 0.01], but not after adjustment for all confounders including high-sensitive C-reactive protein (HR 1.02 per 10 nmol/L, 95% CI 0.87-1.20; P = 0.80). At the same time, hepcidin-25 was significantly related to fatal and non-fatal CV events in a fully adjusted model (HR 1.24 per 10 nmol/L, 95% CI 1.05-1.46, P = 0.01). CONCLUSION: Hepcidin-25 was associated with fatal and non-fatal CV events, even after adjustment for inflammation. Furthermore, inflammation appears to be a significant confounder in the relation between hepcidin-25 and all-cause mortality. These findings suggest that hepcidin-25 might be a novel determinant of CV disease in chronic HD patients.

Background Blood donors that meet the hemoglobin (Hb) criteria for donation may have undetected subclinical iron deficiency. The aim of this study was to assess the prevalence of subclinical iron deficiency in whole blood donors with Hb levels above cutoff levels for donation by measuring zinc protoporphyrin (ZPP) levels. In addition, prevalence rates based on other iron variables were assessed for comparison. Study Design and Methods The study population comprised 5280 Dutch whole blood donors, who passed the Hb criteria for donation. During donor screening, Hb levels were measured in capillary samples (finger prick), and venous blood samples were taken for measurements of ZPP and other iron variables. These variables included ferritin, transferrin saturation, soluble transferrin receptor (sTfR), hepcidin, red blood cell mean corpuscular volume (MCV), and mean cell Hb (MCH). Results With a ZPP cutoff level of at least 100 μmol/mol heme, subclinical iron deficiency was present in 6.9% of male donors and in 9.8% of female donors. Based on other iron variables, iron deficiency was also observed. Prevalence rates ranged from 4.8% (based on transferrin saturation) to 27.4% (based on hepcidin concentration) in men and from 5.6% (based on sTfR concentration) to 24.7% (based on hepcidin concentration) in women. Conclusion Results from this study showed that subclinical iron deficiency is prevalent among blood donors that meet the Hb criteria for blood donation, based on ZPP levels and on other iron variables. This finding needs attention because these donors are at increased risk of developing iron deficiency affecting Hb formation and other cellular processes.

Item does not contain fulltext The effect of carbohydrate (CHO) consumption during prolonged endurance running on post-exercise inflammation and hepcidin levels was investigated. Eleven well-trained male endurance athletes completed a graded exercise test, followed by two experimental running trials in a randomized order. The two experimental trials consisted of a 90 min run at 75% of the peak oxygen uptake velocity (vVO(2peak)), while consuming a solution with either 6% CHO or a placebo (PLA) equivalent at 3 ml kg(-1) every 20 min. Serum interleukin-6 (IL-6), free hemoglobin (Hb), haptoglobin (Hp), hepcidin and iron parameters were assessed throughout the post-run recovery period. Serum iron and IL-6 were significantly elevated immediately post-run in both CHO and PLA (p

Contains fulltext : 80562.pdf (Publisher’s version ) (Open Access) CONTEXT: Obesity and iron deficiency are two of the most common nutritional disorders worldwide. Several studies found higher rates of iron deficiency in obese than in normal-weight children. Hepcidin represents the main inhibitor of intestinal iron absorption, and its expression is increased in adipose tissue of obese patients. Leptin is able, in vitro, to raise hepcidin expression. OBJECTIVES: Aims of this work were 1) to assess the association between poor iron status and obesity, 2) to investigate whether iron homeostasis of obese children may be modulated by serum hepcidin variations, and 3) to assess the potential correlation between leptin and serum hepcidin variations. METHODS: Iron status and absorption as well as hepcidin, leptin, and IL-6 levels were studied in 60 obese children and in 50 controls. RESULTS: Obese children showed lower iron and transferrin saturation (both P < 0.05) and higher hepcidin levels (P = 0.004) compared with controls. A direct correlation between hepcidin and obesity degree (P = 0.0015), and inverse correlations between hepcidin and iron (P = 0.04), hepcidin and transferrin saturation (P = 0.005), and hepcidin and iron absorption (P = 0.003) were observed. A correlation between leptin and hepcidin (P = 0.006) has been found. The correlation remained significant when adjusted for body mass index, sex, pubertal stage, and IL-6 values. CONCLUSIONS: We propose that in obese patients, increased hepcidin production, at least partly leptin mediated, represents the missing link between obesity and disrupted iron metabolism.

Contains fulltext : 69186.pdf (Publisher’s version ) (Closed access) We describe a novel heterozygous mutation in exon 3 of the HFE-gene that was co-inherited with Cys282Tyr in two unrelated Dutch men both presenting a classical form of hereditary hemochromatosis. Heterozygosity for this mutation was also found in one out of 100 healthy controls of Dutch descent. This c.548T>C mutation converts a leucine to a proline residue at position 183 in the alpha2-helix of the HFE-protein (Leu183Pro). Standard bioinformatics analysis shows that the mutation is likely to disturb the HFE interaction with TfR1. This disrupting role of the mutation in the iron regulatory pathway is further corroborated by the familial co-occurrence of the observed compound heterozygosity with increased serum iron parameters. Haplotype analysis strongly suggests that this novel mutation arose from a common ancestor in the distant past. These findings may have implications for HFE-testing of iron overloaded heterozygous Cys282Tyr-patients of Northern European origin and their relatives.

Background Stunting, anemia and inflammation are frequently observed in children with end-stage renal disease (ESRD). Objectives To assess anthropometric, hematological and inflammatory data and to study their potential interrelationship in Guatemalan juveniles undergoing hemodialysis (HD) and peritoneal dialysis (PD). Methods 54 juveniles 7–20 years of age were recruited in FUNDANIER, Guatemala City: 27 on HD and 27 PD. Hemoglobin, serum iron, transferrin, serum transferrin receptor (sTfR), serum ferritin, transferrin saturation and iron-binding capacity, white blood cell count (WBC), erythrocyte sedimentation rate (ESR), C-reactive protein (CRP), as well as IL-6, IL-1 and TNF-α, weight and height were determined by standard methods. Hepcidin–25 (Hep-25) was assessed by weak cation exchange time-of-flight mass-spectrometry. Results 92% and 55% of HD and PD children, respectively, were stunted and 95% and 85% were anemic. Among iron status biomarkers, serum ferritin was massively increased and significantly higher in the HD group compared to the PD group. Hep-25 was also greatly elevated in both groups. 41% of HD patients showed increments in three or more inflammatory biomarkers, while it was 2 or less in all PD subjects. Conclusions The degree of stunting, the prevalence and severity of anemia in Guatemalan juvenile ESRD far exceed the national statistics for this low-income Central American country. Ferritin and Hep-25 concentrations were elevated, with the latter to an extraordinary magnitude. Additional biomarkers of inflammation not directly related to iron status were elevated as well. The role of both disease- and environment-related factors in combination best explains the magnitude of the biomarker abnormalities.

Fetal DNA in plasma and serum of pregnant women has been reported to be significantly increased in preeclampsia (1)(2). This increase may even precede clinical diagnosis (3). We hypothesized that subsequent development of hemolysis, elevated liver enzymes, and low platelet count (HELLP) syndrome will further increase fetal DNA in maternal plasma and serum. In addition, total cell-free DNA may be increased in plasma as a result of tissue destruction, which generally occurs in HELLP syndrome. To confirm our hypothesis, we recruited three groups of pregnant women with informed consent and matched for maternal age [(MA), in years] and gestational age [(GA), in weeks+days] at the time of blood drawing; the median ages (and ranges) were as follows: normotensive women (n = 10), MA, 30 (24–37) and GA, 32+6 (27+0−34+3); preeclamptic women without HELLP syndrome (n = 7), MA, 28 (24–36) and GA, 33+0 (27+6−34+3); and preeclamptic women with HELLP syndrome (n = 10), MA, 31 (26–38) and GA, 26+6−33+6. All women in the latter group had HELLP syndrome at the time of the blood drawing, except two who developed HELLP only after delivery. All participants were carrying single male fetuses. Preeclampsia was defined as a diastolic blood pressure >90 mmHg on two or more consecutive occasions after 20 weeks of pregnancy in previously normotensive women, with proteinuria >0.3 g/L in a 24-h collection period. HELLP syndrome was defined as the simultaneous occurrence of hemolysis (lactate dehydrogenase, >600 U/L), increased liver enzymes (serum aspartate aminotransferase and alanine aminotransferase >70 U/L), and low platelets (

Hepcidin, the key hormone in iron homeostasis,[1][1] is regulated by erythropoietic activity and hypoxia (negative feedback),[2][2] as well as iron stores (hepcidin induction by iron) and inflammation (upregulation).[1][1],[3][3] Only a few publications have focused on hepcidin in the context of

Item does not contain fulltext Abstract Background: Iron deficiency is a commonly encountered problem in pregnancy and a frequently observed cause of pregnancy-associated anemia. We longitudinally assessed the iron regulatory hormone hepcidin during gestation and postpartum and related hepcidin to conventional indicators of iron status and inflammation. Methods: Thirty-one healthy pregnant women were included and 81 blood samples from the three trimesters, directly and 6 weeks postpartum were analyzed for hemoglobin, the iron parameters: iron, total iron binding capacity, transferrin saturation, ferritin, soluble transferrin receptor and hepcidin, and CRP and leucocytes as markers of inflammation. Results: Hepcidin concentration decreased gradually from the first to the second and third trimester to undetectable levels (

Contains fulltext : 125600.pdf (Publisher’s version ) (Open Access) INTRODUCTION: A reliable diagnostic biomarker of iron status is required for severely anemic children living in malarious areas because presumptive treatment with iron may increase their infection risk if they are not iron deficient. Current biomarkers are limited because they are altered by host inflammation. In this study hepcidin concentrations were assessed in severely anemic children living in a highly malarious area of Malawi and evaluated against bone marrow iron in order to determine the usefulness of hepcidin as a point of care test. METHODS: 207 severely anemic children were assessed for levels of hepcidin, ferritin, serum transferrin receptor, erythropoietin, hematological indices, C-reactive protein, interleukin-6, malaria parasites and HIV infection. Deficiency of bone marrow iron stores was graded and erythroblast iron incorporation estimated. Interaction of covariates was assessed by structural-equation-modeling. RESULTS AND CONCLUSION: Hepcidin was a poor predictor of bone marrow iron deficiency (sensitivity 66.7%; specificity 48.5%), and of iron incorporation (sensitivity 54.2%; specificity 61.8%), and therefore would have limitations as a point of care test in this category of children. As upregulation of hepcidin by inflammation and iron status was blunted by erythropoietin in this population, enhanced iron absorption through the low hepcidin values may increase infection risk. Current recommendations to treat all severely anemic children living in malarious areas with iron should therefore be reconsidered.

Contains fulltext : 125929.pdf (Publisher’s version ) (Open Access) Mass spectrometry (MS)-based assays for the quantification of the iron regulatory hormone hepcidin are pivotal to discriminate between the bioactive 25-amino acid form that can effectively block the sole iron transporter ferroportin and other naturally occurring smaller isoforms without a known role in iron metabolism. Here we describe the design, validation and use of a novel stable hepcidin-25(+40) isotope as internal standard for quantification. Importantly, the relative large mass shift of 40 Da makes this isotope also suitable for easy-to-use medium resolution linear time-of-flight (TOF) platforms. As expected, implementation of hepcidin-25(+40) as internal standard in our weak cation exchange (WCX) TOF MS method yielded very low inter/intra run coefficients of variation. Surprisingly, however, in samples from kidney disease patients, we detected a novel peak (m/z 2673.9) with low intensity that could be identified as hepcidin-24 and had previously remained unnoticed due to peak interference with the formerly used internal standard. Using a cell-based bioassay it was shown that synthetic hepcidin-24 was, like the -22 and -20 isoforms, a significantly less potent inducer of ferroportin degradation than hepcidin-25. During prolonged storage of plasma at room temperature, we observed that a decrease in plasma hepcidin-25 was paralleled by an increase in the levels of the hepcidin-24, -22 and -20 isoforms. This provides first evidence that all determinants for the conversion of hepcidin-25 to smaller inactive isoforms are present in the circulation, which may contribute to the functional suppression of hepcidin-25, that is significantly elevated in patients with renal impairment. The present update of our hepcidin TOF MS assay together with improved insights in the source and preparation of the internal standard, and sample stability will further improve our understanding of circulating hepcidin and pave the way towards further optimization and standardization of plasma hepcidin assays.

BACKGROUND The iron-regulating hormone hepcidin is a promising biomarker in the diagnosis of iron disorders. Concentrations of hepcidin have been shown to increase during the day in individuals who are following a regular diet. It is currently unknown whether these increases are determined by an innate rhythm or by other factors. We aimed to assess the effect of dietary iron on hepcidin concentrations during the day. METHODS Within a 7-day interval, 32 volunteers received an iron-deficient diet on 1 day and the same diet supplemented with 65 mg ferrous fumarate at 0815 and 1145 on another day. Blood was drawn to assess ferritin, hepcidin-25, and transferrin saturation (TS) throughout both days at 4 time points between 0800 (fasted) and 1600. A linear mixed model for repeated data was used to analyze the effect of iron intake on TS and hepcidin concentrations. RESULTS Baseline values of hepcidin at 0800 correlated significantly with ferritin (r = 0.61). During the day of an iron-deficient diet the mean TS was similar both in men and in women, whereas hepcidin increased. During the day with iron supplementation the mean TS was significantly higher both in men and in women, and the mean hepcidin was moderately but significantly higher in women (1.0 nmol/L, 95% CI, 0.2–1.8) but not in men (0.0 nmol/L, 95% CI, −0.8 to 0.8). CONCLUSIONS Our data demonstrate that ferritin sets the basal hepcidin concentrations and suggest that innate diurnal rhythm rather than dietary iron mediates the daily hepcidin variations. These findings will be useful for optimizing sampling protocols and will facilitate the interpretation of hepcidin as an iron biomarker.

Contains fulltext : 117437.pdf (Publisher’s version ) (Open Access) Hepcidin, an important regulator of iron homeostasis, is suggested to be causally related to anemia of inflammation. The aim of this study was to explore the role of plasma hepcidin in anemia among older persons from the general population. The Leiden 85-Plus Study is a population-based study of 85-year olds in Leiden, the Netherlands. Eighty-five-year old inhabitants of Leiden were enrolled between September 1997 and September 1999. At the age of 86, plasma hepcidin was determined with time of flight mass spectrometry in 490 participants [160 (32.7%) male, 114 (23.3%) with anemia]. Anemia was defined according to criteria of the World Health Organization (hemoglobin level

Blood donors that meet the hemoglobin (Hb) criteria for donation may have undetected subclinical iron deficiency. The aim of this study was to assess the prevalence of subclinical iron deficiency in whole blood donors with Hb levels above cutoff levels for donation by measuring zinc protoporphyrin (ZPP) levels. In addition, prevalence rates based on other iron variables were assessed for comparison.The study population comprised 5280 Dutch whole blood donors, who passed the Hb criteria for donation. During donor screening, Hb levels were measured in capillary samples (finger prick), and venous blood samples were taken for measurements of ZPP and other iron variables. These variables included ferritin, transferrin saturation, soluble transferrin receptor (sTfR), hepcidin, red blood cell mean corpuscular volume (MCV), and mean cell Hb (MCH).With a ZPP cutoff level of at least 100 μmol/mol heme, subclinical iron deficiency was present in 6.9% of male donors and in 9.8% of female donors. Based on other iron variables, iron deficiency was also observed. Prevalence rates ranged from 4.8% (based on transferrin saturation) to 27.4% (based on hepcidin concentration) in men and from 5.6% (based on sTfR concentration) to 24.7% (based on hepcidin concentration) in women.Results from this study showed that subclinical iron deficiency is prevalent among blood donors that meet the Hb criteria for blood donation, based on ZPP levels and on other iron variables. This finding needs attention because these donors are at increased risk of developing iron deficiency affecting Hb formation and other cellular processes.

Contains fulltext : 98043.pdf (Publisher’s version ) (Closed access) In malaria-endemic areas, iron deficiency and placental Plasmodium falciparum infection commonly coexist. In primigravidae and their newborns, hepcidin and other iron parameters were evaluated in groups and classified according to placental P. falciparum and maternal anemia status. Mothers had relatively high hepcidin levels considering their low iron status. In cord blood, levels of hepcidin, hemoglobin, and other iron parameters were also similar for groups. We conclude that maternal hepcidin is not significantly altered as a function of placental infection and/or anemia. Importantly, fetal hemoglobin and iron status were also unaffected, regardless of the presence of placental infection or maternal anemia.

Contains fulltext : 95951.pdf (Publisher’s version ) (Closed access) Anemia is a common finding among patients with chronic heart failure (HF). Although comorbidities, such as kidney failure, might contribute to the pathogenesis of anemia, many patients with HF do not have any other obvious etiology for their anemia. We investigated whether anemia in HF is associated with an elevation in hepcidin concentration. We used time-of-flight mass spectrometry to measure hepcidin concentration in urine and serum samples of patients with HF and in control subjects. We found that the concentration of hepcidin was lower in urine samples of patients with HF compared with those of control subjects. Serum hepcidin was also reduced in HF but was not significantly lower than that in controls. There were no significant differences between hepcidin levels in patients with HF and anemia compared with patients with HF and normal hemoglobin level. We concluded that hepcidin probably does not play a major role in pathogenesis of anemia in patients with chronic HF.

Contains fulltext : 87990tjalsma.pdf (Publisher’s version ) (Closed access) BACKGROUND: Iron deficiency anemia (IDA) affects many young women in sub-Saharan Africa. Its etiology is multifactorial, but the major cause is low dietary iron bioavailability exacerbated by parasitic infections such as malaria. OBJECTIVE: We investigated whether asymptomatic Plasmodium falciparum parasitemia in Beninese women would impair absorption of dietary iron or utilization of circulating iron. DESIGN: Iron absorption and utilization from an iron-fortified sorghum-based meal were estimated by using oral and intravenous isotope labels in 23 afebrile women with a positive malaria smear (asexual P. falciparum parasitemia; > 500 parasites/muL blood). The women were studied while infected, treated, and then restudied 10 d after treatment. Iron status, hepcidin, and inflammation indexes were measured before and after treatment. RESULTS: Treatment reduced low-grade inflammation, as reflected by decreases in serum ferritin, C-reactive protein, interleukin-6, interleukin-8, and interleukin-10 (P < 0.05); this was accompanied by a reduction in median serum hepcidin of approximately 50%, from 2.7 to 1.4 nmol/L (P < 0.005). Treatment decreased serum erythropoietin and growth differentiation factor 15 (P < 0.05). Clearance of parasitemia increased geometric mean dietary iron absorption (from 10.2% to 17.6%; P = 0.008) but did not affect systemic iron utilization (85.0% compared with 83.1%; NS). CONCLUSIONS: Dietary iron absorption is reduced by approximately 40% in asymptomatic P. falciparum parasitemia, likely because of low-grade inflammation and its modulation of circulating hepcidin. Because asymptomatic parasitemia has a protracted course and is very common in malarial areas, this effect may contribute to IDA and blunt the efficacy of iron supplementation and fortification programs. This trial was registered at clinicaltrials.gov as NCT01108939. 01 december 2010

Contains fulltext : 81093.pdf (Publisher’s version ) (Closed access) PURPOSE: This investigation assessed the effects of training intensity and ground surface type on hemolysis, inflammation, and hepcidin activity during running. METHODS: Ten highly trained male endurance athletes completed a graded exercise test, two continuous 10-km runs on a grass (GRASS) and a bitumen road surface (ROAD) at 75%-80% peak VO2 running velocity, and a 10 x 1-km interval running session (INT) at 90%-95% of the peak VO2 running velocity. Venous blood and urine samples were collected before, immediately after, and at 3 and 24 h after exercise. Serum samples were analyzed for circulating levels of IL-6, free hemoglobin (Hb), haptoglobin (Hp), iron, and ferritin. Urine samples were analyzed for changes in hepcidin expression. RESULTS: After running, the IL-6 and free Hb were significantly greater, and serum Hp was significantly lower than preexercise values in all three conditions (P < 0.05). Furthermore, IL-6 levels and the change in free Hb from baseline were significantly greater in the INT compared with those in the GRASS (P < 0.05). There were no differences between the GRASS and ROAD training surfaces (P > 0.05). Serum iron and ferritin were significantly increased after exercise in all three conditions (P < 0.05) but were not different between trials. CONCLUSION: Greater running intensities incur more inflammation and hemolysis, but these variables were not affected by the surface type trained upon.

Contains fulltext : 79569.pdf (Publisher’s version ) (Closed access) The effect of two running sessions completed within a 12-h period on hemolysis, inflammation, and hepcidin activity in endurance athletes was investigated. Ten males completed two experimental trials in a randomized, counterbalanced order. The two trials included (a) a one-running-session trial (T1) including 10 x 1 km interval repeats (90% peak VO2 velocity), and (b) a two-running-session trial (T2), comprising a continuous 10-km run (70% peak VO2 velocity), and a 10 x 1 km interval run (90% peak VO2 velocity) completed 12 h later. Interleukin-6 (IL-6), free hemoglobin (Hb), haptoglobin (Hp), iron, ferritin, and hepcidin were assessed post-exercise. After the T1 and T2 interval runs, free Hb was significantly increased and Hp significantly decreased (p

Background: Results in previous qualitative studies of the association of the apoptosis inhibitor survivin with prognosis of breast cancer patients have been contradictory. Methods: Survivin mRNA was measured by quantitative TaqMan reverse transcription-PCR in 275 breast cancer tissues from patients with operable tumors and was correlated with established clinicopathologic factors, relapse-free survival [(RFS); 102 events], and overall survival [(OS); 81 events]. Results: High survivin mRNA concentrations were found mainly in tissues from younger patients and in high-grade cancer tissues. High survivin concentrations were most strongly associated with estrogen receptor- or progesterone receptor-negative tumors. In univariate Cox regression analysis for RFS, survivin concentrations were significantly associated with poor prognosis with a hazard ratio (HR) of 1.99 (95% confidence interval, 1.31–3.02; P = 0.001) for every 10-fold increase in expression. For OS, a significant contribution of survivin to poor prognosis was found with a HR of 2.76 (1.67–4.55; P Conclusion: Survivin demonstrates a strong, independent, association with poor prognosis. Survivin might be used as a new marker to stratify breast cancer patients for more optimal treatment modalities, or it could be a promising new target for therapy.

Recently, qualitative analysis of cell-free DNA in blood plasma has attracted much interest for the diagnosis of cancer, fetal gender, Rhesus D status, and inherited disorders (1)(2). Other studies have shown that quantification of total plasma DNA may indicate fetal chromosomal aneuploidies and pregnancy-associated complications or the presence/recurrence of cancer (1)(3)(4). Irrespective of the type of study (qualitative or quantitative DNA analysis), it is important that cell-free plasma DNA is not contaminated with cellular DNA that interferes with analysis and accurate quantification. To prevent cellular DNA contamination, Chiu et al. (5) emphasized the need of preanalytical standardization of blood-processing protocols. The authors note that after centrifugation of blood at a low speed (800g), the amount of isolated DNA from plasma is affected by the presence of cells that remain in the plasma fraction. An additional centrifugation step (16 000g) or filtering of the plasma is necessary to produce absolutely cell-free plasma DNA. Because most laboratories, including ours, centrifuge blood at relatively low speeds (800–1500g) and store plasma without additional treatment, these results may have a serious impact on the usefulness of previously collected plasma samples for retrospective DNA analysis/quantification. To confirm the results of Chiu et al. (5), we collected EDTA blood in 7-mL Vacutainer Tubes from 18 healthy …

Item does not contain fulltext Novel fluorescent oligonucleotides that contain a 3' minor groove binding group (MGB) hybridize to single-stranded targets with increased sequence-specificity compared to ordinary DNA probes. This reduces non-specific probe hybridization and results in low background fluorescence during the 5' nuclease PCR assay (TaqMan, Applied Biosystems, Foster City, CA). We developed a method for closed-tube genotyping using two allele-specific MGB probes labeled with different fluorophores in one reaction. After PCR, tubes were transported to a fluorescence plate-reader for analysis of fluorescence. Common spreadsheet software was used for automated genotype assignment. As an example, DNA samples from 172 hemochromatosis patients were selected and tested for molecular defects in the HFE gene, i.e., mutations in codon 63 and 282. Tight genotype clusters were observed for both codons and results with MGB probes were identical to conventional genotyping (PCR + restriction-fragment-length-polymorphism). We show that this fast and easy method can be used for large-scale (high-throughput) genetic studies but also for routine molecular diagnostics without post-PCR manipulation of amplicons or the need for real-time quantitative PCR machines. Hum Mutat 19:554-559, 2002.

Contains fulltext : 137963.pdf (Publisher’s version ) (Open Access) BACKGROUND: This investigation compared the effects of an extended period of weight-bearing (running) vs. non-weight-bearing (cycling) exercise on hepcidin production and its implications for iron status. METHODS: Ten active males performed two separate exercise training blocks with either running (RTB) or cycling (CTB) as the exercise mode. Each block consisted of five training sessions (Day 1, 2, 4, 5, 6) performed over a seven day period that were matched for exercise intensity. Basal venous blood samples were obtained on Day 1 (D1), and on Recovery Days 3 (R3) and 7 (R7) to assess iron status, while basal and 3 h post-exercise urinary hepcidin levels were measured on D1, D2, D6, as well as R3 and R7 (basal levels only) for each condition. RESULTS: Basal urinary hepcidin levels were significantly elevated (p

To the editor: We present a new homozygous truncating mutation, L165X, of the hemojuvelin gene ( HJV ), observed in one male patient with severe juvenile hemochromatosis (JH). Because the C282Y-variant in the hemochromatosis gene ( HFE ) was also common in this family, we investigated whether the

Urinary hepcidin, inflammation, and iron metabolism were examined during the 24 hr after exercise. Eight moderately trained athletes (6 men, 2 women) completed a 60-min running trial (15-min warm-up at 75–80% HRpeak + 45 min at 85–90% HRpeak) and a 60-min trial of seated rest in a randomized, crossover design. Venous blood and urine samples were collected pretrial, immediately posttrial, and at 3, 6, and 24 hr posttrial. Samples were analyzed for interleukin-6 (IL-6), C-reactive protein (CRP), serum iron, serum ferritin, and urinary hepcidin. The immediate postrun levels of IL-6 and 24-hr postrun levels of CRP were significantly increased from baseline (6.9 and 2.6 times greater, respectively) and when compared with the rest trial (p ≤ .05). Hepcidin levels in the run trial after 3, 6, and 24 hr of recovery were significantly greater (1.7–3.1 times) than the pre- and immediate postrun levels (p ≤ .05). This outcome was consistent in all participants, despite marked variation in the magnitude of rise. In addition, the 3-hr postrun levels of hepcidin were significantly greater than at 3 hr in the rest trial (3.0 times greater, p ≤ .05). Hepcidin levels continued to increase at 6 hr postrun but failed to significantly differ from the rest trial (p = .071), possibly because of diurnal influence. Finally, serum iron levels were significantly increased immediately postrun (1.3 times, p ≤ .05). The authors concluded that high-intensity exercise was responsible for a significant increase in hepcidin levels subsequent to a significant increase in IL-6 and serum iron.