Tissue Iron Stores Research Articles

Association of Cord Ferritin Levels with Brain Stem Evoked Response Audiometry in Newborns.

Iron is an important nutrient and it plays a pivotal role in myelin formation and neurotransmitter synthesis, thus contributing to normal neurological activity. Ferritin is a reliable indicator of the tissue iron stores and in-utero stores can be well measured by cord ferritin levels. The objective of this study was to evaluate the effects of umbilical cord ferritin levels (CFL) on the brain stem evoked response audiometry (BERA) This prospective observational study was conducted in a tertiary care centre of North India with a sample size of 100 inborn neonates. After evaluation of the umbilical cord ferritin levels the study cohort was divided into Group A( UCF<75ng/ml) and Group B(UCF>75ng/dl). All subjects were subjected to BERA. A detailed analysis of CFL and BERA was done and statistically analysed. Neonates in group B had significantly prolonged absolute peak latency of wave I, III and V and interpeak latency of wave III-V when compared to group A. The Pearson correlation also showed negative correlation of CFL with absolute peak latency of wave I, III, and V and interpeak latency of wave III-V. Among the maternal and neonatal variables, highly significant correlation was noted between absolute latency of wave I, III and V, CFL and cord hematocrit. The Pearson correlation showed negative correlation of absolute peak latency of wave I, III and V with maternal haemoglobin (Hb), neonatal birth weight, CFL,s and cord hematocrit values. A negative Pearson correlation was noted between interpeak latency of wave III-V with neonatal birth weight and cord hematocrit level, interpeak latency of wave I-V with neonatal birth weight and between interpeak latency of wave I-III with cord hematocrit values. CFL's significantly affect the absolute peak latency of wave I, III and V and it also affects the interpeak latency of wave III-V. This may be attributed to slow conduction time secondary to altered myelination. CFL's should be considered as a routine protocol in neonates to detect early compromises in the process of myelination and brain maturation.

Математическое прогнозирование рисков железодефицитных состояний у женщин

Purpose. Based on the study of medico-social, medico-biological and psychological factors, to assess the risks of iron deficiency in women. The results obtained will be used to develop a relevant program for the prevention of iron deficiency anemia and latent iron deficiency in primary health care. Materials and methods. The medical and social study was conducted on the basis of the city polyclinics of Kazan, the samples included two groups of women: 394 patients with laboratory-confirmed iron deficiency, including its clinical manifestations, and 373 women with a confirmed absence of iron deficiency and without probable complaints. The following methods were used in the study: bibliographic, socio-hygienic, psychological testing, sociological, information-analytical, mathematical-statistical, organizational experiment. Results. New data have been established on the factors affecting the volume of tissue iron stores in women: statistically significant associations of serum ferritin with sleep quality parameters and carbohydrate intake. The risks of iron deficiency anemia and latent iron deficiency were assessed, and subranges for individual risk groups were calculated. Findings. Statistically significant associations were found in the ferritin-total protein model with late bedtime (p &lt;0,001), discrete sleep patterns (p &lt;0,001), situational anxiety (p &lt;0,001), and daily sugar intake (p = 0,021). According to mathematical forecasting, the greatest influence on the development of IDA in women is “going to bed” (R=2,1). Women who go to bed after midnight and from 23.00 to midnight are at risk of iron deficiency anemia, the risk of which is 2 times higher than in women who fall asleep before 23.00. The probability of latent iron deficiency in women with a high level of personal and situational anxiety is 28–29 times higher than in women with a low level of anxiety (R=29,254 and R=28, respectively).

An Experimental Model for Iron Deficiency Anemia in Sows and Offspring Induced by Blood Removal during Gestation.

Simple SummaryAnemia is a common condition in sow herds especially during gestation. The condition occurs despite iron supplementation in feed and has been associated with increased stillbirth rate. We currently have limited knowledge about the consequences of anemia during gestation in sows. The aim of this study was to assess the effects of severe iron deficiency during gestation on sow and piglet health outcomes in an experimental model. For sows, hematologic parameters were monitored at regular intervals and farrowing results were recorded. After farrowing, tissue iron concentrations in sows and new born piglets were measured. Results of our study showed that sows and piglets were adversely affected by iron deficiency anemia (IDA) during gestation. Mild to severe IDA resulted in reduced tissue iron stores at farrowing in sows and piglets. Strategies to overcome the condition could therefore be beneficial to the pig industry.Anemia is a common condition in sow herds. We aimed to study the effects of severe iron deficiency during gestation on sow and piglet health outcomes with an experimental model for blood-removal-induced iron deficiency anemia. In total, 18 multiparous sows (8 in trial I and 10 in trial II) were allocated to either a blood removal group or a control group. Hematologic parameters were monitored at regular intervals and the tissue iron concentrations were measured for the sows and newborn piglets after farrowing. In trial I, the mean liver iron content was reduced to 46.7 µg/g in the blood removal sows compared to 252.6 µg/g in the controls (p < 0.001). In trial II, sows in the blood removal group had lower iron content in the liver (147.8 µg/g), kidney (46.3 µg/g) and spleen (326.5 µg/g) compared to the control sows (323.2 µg/g, 81.3 µg/g and 728.9 µg/g, respectively; p = 0.009, 0.016, 0.01, respectively). In trial I, piglets from sows in the blood removal group had significantly decreased hematocrit (Hct), red blood cells (RBC) and a tendency for reduced hemoglobin (Hb) compared to the control piglets. We established a blood removal model that resulted in mild- to severe degrees of sow anemia and reduced tissue iron stores at farrowing.

Characterization of Iron Metabolism in Mouse Model of Congenital Polycythemia with Human Gain-of-Function Erythropoietin Receptor

Hepcidin (encoded by HAMP), the peptide hormone regulating systemic iron homeostasis, controls iron flux to plasma and thus ensures adequate iron supply for erythropoiesis. Increased erythropoietic activity negatively regulates hepcidin synthesis by the hormone erythroferrone (ERFE) produced by erythroid precursors in response to erythropoietin (EPO). Here we evaluated iron metabolism in a mouse model of polycythemia with augmented erythropoiesis and low EPO levels bearing the gain-of-function polycythemia-causing human EPO receptor gene (mtHEPOR) (Divoky PNAS 2002). The mtHEPOR embryos developed polycythemia around embryonic day (ED) 17.5, followed by unexpected but transient correction of polycythemia in perinatal life (postnatal day 7 - PD7) and reappearance of a polycythemic phenotype at 3-6 weeks of age. (Human patients with the gain-of-function polycythemia-causing human EPOR mutations are also born non-polycythemic and then develop polycythemia within a few weeks of neonatal life). The mtHEPOR mice had reduced Epo levels, augmented EPOR signaling and prolonged activation of Stat5 in erythroblasts when compared to mEpoR mice (Divoky JMolMed 2016). To assess whether developmental changes of erythron in this mouse model correlate with changes in iron metabolism, we determined temporal changes of Erfe and Hamp expression and body iron stores.We found that mtHEPOR mice have increased expression of Erfe at ED 17.5 (7.5-times) and PD7 (4-times) compared to mEpoR controls when normalized to a housekeeping gene beta-actin and independently to erythroid marker glycophorin A. Therefore Erfe in mtHEPOR mice would be increased both because of a higher number of erythroid precursors and because each precursor is producing more Erfe by increased transcription. This corresponds to reduced expression of Hamp in the liver of ED17.5 (4-times) and PD7 mtHEPOR mice (15-times) compared to mEpoR littermates. However, perinatally (PD1), Erfe expression dramatically decreased in mtHEPOR mice (7-times) concomitantly with increased Hamp expression (45-times), and both these factors reached expression levels detected in mEpoR at PD1. Expression of Erfe and Hamp between mtHEPOR and mEpoR mice was also not different in older mice (7 months of age), even though mtHEPOR mice were markedly polycythemic with persistent erythropoietic activity, markedly increased splenic erythropoiesis (the weight of spleens of control animals was 100-180 mg compared to 200-350 mg of mtHEPOR mice), and augmented EPOR-Stat5 signaling. However, mtHEPOR and mEpoR mice differed in transferrin saturation (TSAT) and tissue iron stores. The spleens of mtHEPOR mice had markedly increased iron deposits compared to mEpoR controls and stainable iron could only be detected in the livers of mtHEPOR mice but not in the livers of mEpoR controls. TSAT was elevated in mtHEPOR mice (77±3.2%) when compared to mEpoR (54.7±8.0%), consistent with the iron overload in mtHEPOR mice. Nevertheless, Hamp expression was comparable between mtHEPOR and mEpoR mice. This shows that Hamp expression was unexpectedly normal in respect to the presence of iron overload in mtHEPOR mice at 7 months of age.With further aging (mice 14 months and older), Erfe continued to increase and Hamp continued to decrease in mEpoR mice. In contrast, there was a trend towards decreased Erfe expression while Hamp expression became significantly elevated (2-times) in mtHEPOR mice. These changes were accompanied by a 2.4-fold decline in TSAT in both genotypes, which suggests that aged mEpoR mice develop iron deficiency while mtHEPOR mice remain iron-replete likely due to mobilization of iron from high iron stores.We show that the polycythemic phenotype, as well as Erfe and Hamp expression, in mtHEPOR mice undergoes dynamic changes during ontogenesis. We conclude that further research will be needed to understand the development of iron overload and changes in Erfe and Hamp expression in mtHEPOR mice characterized by erythroid hyperplasia and low Epo levels.Acknowledgment: BK, VD, and MH were supported by the Czech Science Foundation (projects GA15-13732S and GA17-05988S), KONTAKTII project LH15223, and in part by project LTAUSA17142. BK and JS contributed equally to this work. JTP and MH act as equivalent co-senior authors. DisclosuresNo relevant conflicts of interest to declare.

2315-PUB: High Serum Ferritin Predicts Inferior Diabetes-Related Outcomes after Gastric Bypass but Not Sleeve Gastrectomy Bariatric Surgery

Objective: Because higher iron level is associated with increased risk of type 2 diabetes and bariatric surgery is known to alter iron homeostasis, we examined the effects of tissue iron stores on glycemic outcome after bariatric surgery. Research Design and Methods: We performed a retrospective analysis of glycemia outcomes after Roux-en Y gastric bypass (RYGB) or vertical sleeve gastrectomy (VSG) in individuals with elevated hemoglobin A1c (HbA1c) as a function of serum ferritin before and after surgery, using two large and independent electronic health databases from the Carolinas Collaborative and University of Michigan. Results: Improvement of HbA1c, weight, and alanine aminotransferase (ALT) was comparable between the two procedures 6-12 months after surgery. For gastric bypass procedures, however, the improvement in HbA1c and the chance of normalizing HbA1c were inversely proportional to preoperative ferritin, while no such relationship existed for gastrectomy. The improvement in ALT was correlated with the decrease in serum ferritin after surgery for both surgery types. Changes in hemoglobin were also related to preoperative ferritin. Conclusion: Preoperative iron stores may modulate glycemic improvement after bariatric surgery differentially depending on procedure type. This has implications regarding the operation of choice in certain patient populations. Disclosure J. Tong: None. R. Alvarez: None. G.B. Russell: None. A.N. Khouri: None. R.J. Seeley: Consultant; Self; Ionis Pharmaceuticals, Inc., Kintai, Kintai, Novo Nordisk A/S, Sanofi, Scohia Pharma Inc. Research Support; Self; AstraZeneca, Novo Nordisk A/S, Pfizer Inc. Stock/Shareholder; Self; Redesign Health. D.A. McClain: None. Funding National Institutes of Health (R01DK097550)

PF500 A NEW FTL GENE MUTATION RESPONSIBLE FOR HYPOFERRITINEMIA WITHOUT IRON DEFICIENCY

Background:Serum ferritin (sFt), is widely considered the best test to assess iron deficiency, as it reflects tissue iron storage. A decrease in sFt is an early indicator of a lack of iron in the body and precedes the decrease of serum iron and the onset of iron deficiency anemia. We present an exceptional case of hypoferritinemia, without anemia or associated iron deficiency. Repeated injections of intravenous iron to the patient led to severe iron overload.Aims:Case report: A 34‐year‐old woman was referred to our Center for investigation of iron overload. She reported that three years ago a biological assessment to investigate asthenia revealed very low sFt (1 μg/L). The CBC showed no anemia nor microcytosis. The patient had previously received oral iron supplementation for 8 years because of this low sFt. She was given IV iron and received 18 injections in 15 months. Following these injections, ferritin normalized (69 μg/L), but transferrin saturation (TSat) was measured at 96%. The patient consulted a few months later for severe asthenia, anorexia, weight loss and dyspnoea and was hospitalized. Liver iron concentration (LIC) measured by MRI was 190 μmol/g (N &lt;36 μmol/g). The diagnosis of secondary haemochromatosis was given and phlebotomies were started. When we saw the patient for second opinion, she already had 9 phlebotomies. Iron parameters showed a SFt at 2 μg/L, serum iron at 31.48 μmol/L and TSat at 92%. Hemoglobin (Hb) levels were normal at 128 g/L with a MCV at 98 fL. Due to a poor clinical tolerance of the phlebotomies, oral iron chelation was proposed. The patient reported that her father, mother, two sisters, and one brother also had unexplained hypoferritinemia and had received iron supplementation.Methods:After informed written consent, Sanger sequencing of the 5’UTR and exon 1 of the FTL (ferritin light chain) gene and NGS analysis of a panel of genes involved in hereditary iron overload (BMP6, HAMP, HFE, HJV, SLC40A1, TRF2) were performed.Results:An undescribed mutation of the FTL gene (c.92A&gt; G or p.Y31C), was identified at the homozygous state. A mutation of the BMP6 gene (c: 337C&gt; G or p.Gln133Glu) in the heterozygous state was also observed. This already described mutation is associated with a decrease expression of hepcidin and is thought to promote iron overload. However, hepcidin was measured 3 times at normal values in our patient.Summary/Conclusion:The mutation identified in the FTL gene is localized in the region involved in L ferritin secretion. Two hypotheses could account for the hypoferritinemia presented by the patient. It could be a lack of serum secretion of the L‐ferritin chain, despite normal tissue iron stores, or a modification of the recognition site by the antibodies used for ferritin assays (immunological). In order to explore these hypotheses, a study of family segregation and different assay techniques for ferritin were investigated. In addition, the BMP6 gene mutation could have played an aggravating cofactor role, favouring the occurrence of acquired iron overload in this patient. This case illustrates the importance of a pragmatic attitude in the presence of any discrepancies among iron parameters. This isolated hypoferritinemia without anemia or other evidence of iron deficiency, was not an indication of IV iron treatment, which led to iron overload and serious complications in the patient.

Iron Metabolism of the Skeletal Muscle and Neurodegeneration.

Recent studies clearly indicate that the endocrine function of the skeletal muscle is essential for a long and healthy life. Regular exercise, which has been shown to stimulate the release of myokines, lowers the risk of many diseases, including Alzheimer’s and Parkinson’s disease, emphasizing the role of skeletal muscle in proper functioning of other tissues. In addition, exercise increases insulin sensitivity, which may also impact iron metabolism. Even though the role of iron in neurodegeneration is well established, the exact mechanisms of iron toxicity are not known. Interestingly, exercise has been shown to modulate iron metabolism, mainly by reducing body iron stores. Insulin signaling and iron metabolism are interconnected, as high tissue iron stores are associated with insulin resistance, and conversely, impaired insulin signaling may lead to iron accumulation in an affected tissue. Excess iron accumulation in tissue triggers iron-dependent oxidative stress. Further, iron overload in the skeletal muscle not only negatively affects muscle contractility but also might impact its endocrine function, thus possibly affecting the clinical outcome of diseases, including neurodegenerative diseases. In this review, we discuss possible mechanisms of iron dependent oxidative stress in skeletal muscle, its impact on muscle mass and endocrine function, as well as on neurodegeneration processes.

Insulin secretion-sensitivity index-2 could be a novel marker in the identification of the role of pancreatic iron deposition on beta-cell function in thalassemia major

The purpose of this study is to evaluate the impact of insulin secretion-sensitivity index-2 (ISSI-2) in the identification of the role of pancreatic iron deposition on beta-cell function in thalassemia major. Tissue iron stores were measured with magnetic resonance imaging (MRI) in the liver (R2), pancreas (R2*), and heart (T2*). ISSI-2 was assessed as a novel oral glucose tolerance test-based measure of beta-cell function. Also, the Stumvoll index showing the insulin sensitivity and Stumvoll index estimating first and second phase insulin secretion were calculated. Fourteen of the 51 Thalassemia Major patients, aged 8-34 (mean 21.1 ± 7.2) years-old, had either an impaired glucose tolerance test (n = 9, 17.6%) or diabetes mellitus (n = 5, 9.8%)-referred to as the glucose dysregulation (GD) group. The median serum ferritin and the mean liver R2 and cardiac T2* values were not significantly different between the GD and normal glucose tolerance (NGT, n = 37) groups whereas pancreas R2* was significantly higher in the GD group compared to the NGT group (p = 0.004). Patients with GD showed significantly lower ISSI-2 index (p < 0.001) as well as the Stumvoll index and Stumvoll first and second phase indices compared to those with NGT (p < 0.001). All patients with GD displayed a pancreas R2* >50 Hz and ISSI-2 <2. In conclusion, Pancreas R2* MRI combined with ISSI-2 index may be valuable parameters to identify patients at the highest risk for developing glucose dysregulation.

Iron, erythropoietin, and inflammation regulate hepcidin in Bmp2-deficient mice, but serum iron fails to induce hepcidin in Bmp6-deficient mice.

The bone morphogenetic protein (BMP)-SMAD signaling pathway is a key transcriptional regulator of hepcidin in response to tissue iron stores, serum iron, erythropoietic drive and inflammation to increase the iron supply when needed for erythropoiesis, but to prevent the toxicity of iron excess. Recently, BMP2 was reported to play a non-redundant role in hepcidin regulation in addition to BMP6. Here, we used a newly validated BMP2 ELISA assay and mice with a global or endothelial conditional knockout (CKO) of Bmp2 or Bmp6 to examine how BMP2 is regulated and functionally contributes to hepcidin regulation by its major stimuli. Erythropoietin (EPO) did not influence BMP2 expression in control mice, and still suppressed hepcidin in Bmp2 CKO mice. Lipopolysaccharide (LPS) reduced BMP2 expression in control mice, but still induced hepcidin in Bmp2 CKO mice. Chronic dietary iron loading that increased liver iron induced BMP2 expression, whereas acute oral iron gavage that increased serum iron without influencing liver iron did not impact BMP2. However, hepcidin was still induced by both iron loading methods in Bmp2 CKO mice, although the degree of hepcidin induction was blunted relative to control mice. Conversely, acute oral iron gavage failed to induce hepcidin in Bmp6 -/- or CKO mice. Thus, BMP2 has at least a partially redundant role in hepcidin regulation by serum iron, tissue iron, inflammation and erythropoietic drive. In contrast, BMP6 is absolutely required for hepcidin regulation by serum iron.

The novel compound PBT434 prevents iron mediated neurodegeneration and alpha-synuclein toxicity in multiple models of Parkinson\u2019s disease

Elevated iron in the SNpc may play a key role in Parkinson’s disease (PD) neurodegeneration since drug candidates with high iron affinity rescue PD animal models, and one candidate, deferirpone, has shown efficacy recently in a phase two clinical trial. However, strong iron chelators may perturb essential iron metabolism, and it is not yet known whether the damage associated with iron is mediated by a tightly bound (eg ferritin) or lower-affinity, labile, iron pool. Here we report the preclinical characterization of PBT434, a novel quinazolinone compound bearing a moderate affinity metal-binding motif, which is in development for Parkinsonian conditions. In vitro, PBT434 was far less potent than deferiprone or deferoxamine at lowering cellular iron levels, yet was found to inhibit iron-mediated redox activity and iron-mediated aggregation of α-synuclein, a protein that aggregates in the neuropathology. In vivo, PBT434 did not deplete tissue iron stores in normal rodents, yet prevented loss of substantia nigra pars compacta neurons (SNpc), lowered nigral α-synuclein accumulation, and rescued motor performance in mice exposed to the Parkinsonian toxins 6-OHDA and MPTP, and in a transgenic animal model (hA53T α-synuclein) of PD. These improvements were associated with reduced markers of oxidative damage, and increased levels of ferroportin (an iron exporter) and DJ-1. We conclude that compounds designed to target a pool of pathological iron that is not held in high-affinity complexes in the tissue can maintain the survival of SNpc neurons and could be disease-modifying in PD.

Correlation of serum ferritin levels, in female patients with chronic diffuse hair loss: A cross sectional study

Context: Iron is involved in critical physiological processes within the hair follicle, suggesting that iron deficiency (ID) could disrupt hair synthesis. The relationship between body iron status and hair loss has been investigated in a number of studies, however, with relatively discrepant findings. Aim: The aim of this study is to evaluate whether chronic telogen effluvium (CTE) and female pattern hair loss (FPHL) in patients were associated with decreased tissue iron stores, as measured by serum ferritin levels. Setting and Design: This was a cross-sectional study, conducted in a teaching hospital. Materials and Methods: Forty female patients aged between 15 and 45 years, having chronic diffuse hair loss, were recruited. Statistical Analysis Used: Statistical analysis used in this study was Mean ± standard deviation, Pearson's correlation coefficient, and one-sample t-test. Results: The mean serum ferritin levels in all participants, CTE, and FPHL were 18.6 μg/L, 16 μg/L, and 36.64 μg/L, respectively. Almost 57.5% of participants had serum ferritin levels Conclusion: Participants of this study had low serum ferritin levels at different definitions of ID, and the participants showed ID, which was significant. Hence, chronic diffuse hair loss was associated with decreased iron stores.

Bone Mineral Density in Postmenopausal Women Heterozygous for the C282Y HFE Mutation.

Mutations in the HFE gene may be associated with increased tissue iron stores reflected in an elevated serum ferritin. With homozygous mutation C282Y, the increase in serum ferritin may be associated with tissue damage in the liver, pancreas, and pituitary and with a reduced bone mineral density. With heterozygous mutation C282Y, the degree of iron retention is less but information relating to how a heterozygous C282Y mutation might impact bone mineral density is uncertain. The present study was undertaken to study the relationships between bone mineral density measured by dual energy X-ray absorptiometry and the serum ferritin and serum iron in postmenopausal women heterozygous for the C282Y mutation. The spinal bone mineral density, L2–4, was significantly less than age matched community controls (P = 0.016). There was no significant change in the femoral neck bone mineral density compared to age matched community controls. The correlation between the spinal bone mineral density, L2–4, the femoral neck bone mineral density, and the serum ferritin was not significant. The serum iron correlated significantly inversely with the femoral neck bone mineral density (P = 0.048). The heterozygous C282Y mutation may be associated with impairment of bone cell function in postmenopausal women when only small increases in the serum iron or serum ferritin have occurred.

Brain iron overload, insulin resistance, and cognitive performance in obese subjects: a preliminary MRI case-control study.

The linkage among the tissue iron stores, insulin resistance (IR), and cognition remains unclear in the obese population. We aimed to identify the factors that contribute to increased hepatic iron concentration (HIC) and brain iron overload (BIO), as evaluated by MRI, and to evaluate their impact on cognitive performance in obese and nonobese subjects. We prospectively recruited 23 middle-aged obese subjects without diabetes (13 women; age 50.4 ± 7.7 years; BMI 43.7 ± 4.48 kg/m2) and 20 healthy nonobese volunteers (10 women; age 48.8 ± 9.5 years; BMI 24.3 ± 3.54 kg/m2) in whom iron load was assessed in white and gray matter and the liver by MRI. IR was measured from HOMA-IR and an oral glucose tolerance test. A battery of neuropsychological tests was used to evaluate the cognitive performance. Multivariate regression analysis was used to identify the independent associations of BIO and cognitive performance. A significant increase in iron load was detected at the caudate nucleus (P < 0.001), lenticular nucleus (P = 0.004), hypothalamus (P = 0.002), hippocampus (P < 0.001), and liver (P < 0.001) in obese subjects. There was a positive correlation between HIC and BIO at caudate (r = 0.517, P < 0.001), hypothalamus (r = 0.396, P = 0.009), and hippocampus (r = 0.347, P < 0.023). The area under the curve of insulin was independently associated with BIO at the caudate (P = 0.001), hippocampus (P = 0.028), and HIC (P = 0.025). BIOs at the caudate (P = 0.028), hypothalamus (P = 0.006), and lenticular nucleus (P = 0.012) were independently associated with worse cognitive performance. Obesity and IR may contribute to increased HIC and BIO being associated with worse cognitive performance. BIO could be a potentially useful MRI biomarker for IR and obesity-associated cognitive dysfunction.

Cord Blood Serum Ferritin of Infants of Diabetic Mothers

Introduction:Maternal diabetes mellitus is associated with depleted fetal iron stores and this is proportionate to the degree of maternal control, presence or absence of diabetes-related complications, and is not related to maternal iron status. In this study, we aim to assess the effect of maternal diabetes on cord blood serum ferritin. Methods: The present prospective (case-control) study was carried out in AL-Zahraa Teaching Hospital (March 2012-October 2012). Umbilical cord blood samples were collected from 100 newborn infants who were delivered normally or by caesarean section. Fifty infants of diabetic mothers (IDMs) and 50 normal control neonates were randomly recruited. A serum sample was obtained to measure ferritin concentrations by mini VIDAS machine, which compares the results with the standards. Statistical analysis was performed using SPSS (statistical package for social sciences) Version 17. Independent sample t-test was used for data measurement and chi-square test for analyzing the categorical data. Also, Pearson’s correlation coefficient was used to compare two measurement variables. P-valueResults: There was no significant difference between IDMs and infants of healthy mothers, regarding the gestational age at the time of delivery (P=0.31). Also, there was no significant difference between the two groups, regarding their packed cell volume (PCV), mean corpuscular volume (MCV), and red cell distribution width (RDW) (P>0.05). Finally, there was a highly significant difference between the two groups, regarding cord blood serum ferritin (P<0.05). Conclusion: This study shows that IDMs have lower tissue iron stores (S. ferritin) at birth. Also, according to the results, there is a significant association between S. ferritin ,gestational age and birth weight in these neonates.

Transferrin Deficiency Leads To Specific and Partially Reversible Iron Overload

Studies in hpx mice, a model of inherited transferrin deficiency, have demonstrated that transferrin is essential for iron delivery to the erythron and regulation of expression of hepcidin, a hormone that inhibits dietary iron absorption. Here we address two other putative roles for s transferrin. First, we determined if transferrin is essential for metabolism of metals other than iron. Metal content analysis of fractionated mouse sera indicated that iron was the most abundant metal in transferrin-rich fractions. Organ metal content analysis demonstrated severe imbalances for iron and minimal to moderate imbalances for copper, zinc and manganese in hpx tissues. Analysis of metal-dependent gene expression and organ metal content in a mouse model of inherited hepcidin excess suggested that hpx tissue imbalances in copper, zinc and manganese levels were not all physiologic or specific to primary transferrin deficiency. Second, we tested if transferrin modulates the mobilization of tissue iron stores. Transferrin treatment of hpx mice decreased iron levels in most tissues analyzed. Iron mobilization was independent of hepcidin, as iron levels decreased in hpx mice deficient in hemojuvelin, a membrane protein essential for hepcidin expression. Iron mobilization led to decreased ferritin iron and heavy chain levels. The decrease in liver ferritin iron stores did not match the decrease in total liver iron stores, suggesting that iron was mobilized from multiple storage sites. Overall, our analysis suggests that the primary consequences of transferrin deficiency are largely specific to iron and that transferrin can modulate the mobilization of tissue iron stores. Disclosures:No relevant conflicts of interest to declare.

Combined Chelation Therapy with Deferasirox and Deferoxamine In Transfusion-Dependent Thalassemia

AbstractAbstract 4269Therapeutic regimens that combine two iron chelators may enhance chelation efficiency by improving access to different tissue iron stores and control of the toxic labile iron pool. The combination of two chelators can reduce toxicity through averting the need for high doses of a single drug, but it is essential to establish the safety such regimens. We therefore explored the combined use of deferasirox (DSX) and deferoxamine (DFO) in patients with transfusion-dependent thalassemia who had failed standard chelation therapy with single drug. Patients were eligible if the liver iron concentration (LIC) >15 mg/g dry liver-weight or if iron-induced end organ injury was present. Subjects were monitored for hepatic and renal toxicity, visual or auditory changes, and the development of new complications from iron overload. The ability of the combined therapy to control systemic iron burden (serum ferritin and LIC) and myocardial iron overload (MRI T2*) was evaluated. We also measured changes in plasma levels of non-transferrin bound iron (NTBI) and labile plasma iron (LPI).Fifteen subjects were enrolled in 3 groups: adults with LIC <15 mg/g dry liver-weight (group A), adults with LIC >15 mg/g (group B), and children 8–18 years with LIC >5 mg/g (group C). The duration of therapy was 52 weeks. DSX (20-30 mg/Kg) was administered daily and DFO (35-50 mg/Kg/infusion) was infused on 3–7 days/week (as 8–12 hour infusion) based upon the degree of iron overload present at baseline. At the initiation of the study, the mean daily dose of DFO was 16, 33, and 17 mg/Kg/day and mean DSX dose was 21, 25 and 22 mg/Kg/day for groups A, B and C, respectively. At the conclusion of the trial, the median LIC declined by 48% from 10.8 mg/g (3.9-34.8 mg/g) to 5.7 mg/g (1.0-24.0 mg/g, p=0.003). The median ferritin fell by 43% from 2030 ng/mL (1000-5230 ng/mL) to 1150 ng/mL (421-5260 ng/mL, p=0.008). Myocardial iron in the 3 subjects who had T2* <20 msec at study entry (range 6.5–19.5 msec at week 0) showed an average improvement of +2.43 msec following treatment (range 8.8–21.3 msec at week 52, p=0.027). All 3 subjects with left ventricular ejection fraction below 60% at baseline (47.5-58.1%) showed improvement at end of study (60.6-64.4%). There was progressive decline in median plasma NTBI level during the study from 3.26 μM (1.79-5.79 μM) at baseline to 2.38 μM (1.59-3.08 μM) at 12 months (p=0.008). DSX produced immediate and significant decline in plasma NTBI when administered during infusion of DFO. The median plasma NTBI measured on DFO alone was 2.46 μM (0.92-5.90 μM), which decreased to 1.96 μM (0-3.50 μM) following administration of the dose of DSX (p<0.001). A sustained control of the LPI fraction was also demonstrated throughout the study period. At baseline the median LPI was 0.87 μM (0-2.43 μM) which decreased to 0.05 μM (0-1.20 μM) during the study period (p=0.004).No significant toxicity or unusual adverse events were observed with combined chelation therapy in this group of high-risk patients with thalassemia. Elevation of serum creatinine or ALT was not observed in any subject. One subject from group B died at 9 weeks from start of trial from sepsis. One subject interrupted DSX therapy because of abdominal pain. In all other cases the treatment was well tolerated and no dose adjustment or suspension of therapy was required on account of toxicity. Protocol-mandated modification of treatment (temporary cessation of DSX or DFO) occurred in three subjects owing to a marked fall in serum ferritin and LIC. These results suggest that simultaneous administration DSX and DFO is well tolerated and has low potential for toxicity. Combined chelation therapy appears to be effective in rapidly reducing systemic iron burden, lowering myocardial iron, and controlling plasma NTBI and LPI in patients at risk of developing end-organ damage. Disclosures:Harmatz:Ferrokin: Membership on an entity's Board of Directors or advisory committees, Research Funding; Novartis: Research Funding. Porter:Novartis: Membership on an entity's Board of Directors or advisory committees, Research Funding, Speakers Bureau. Vichinsky:Novartis: Membership on an entity's Board of Directors or advisory committees, Research Funding, Speakers Bureau.

Serum ferritin is derived primarily from macrophages through a nonclassical secretory pathway

The serum ferritin concentration is a clinical parameter measured widely for the differential diagnosis of anemia. Its levels increase with elevations of tissue iron stores and with inflammation, but studies on cellular sources of serum ferritin as well as its subunit composition, degree of iron loading and glycosylation have given rise to conflicting results. To gain further understanding of serum ferritin, we have used traditional and modern methodologies to characterize mouse serum ferritin. We find that both splenic macrophages and proximal tubule cells of the kidney are possible cellular sources for serum ferritin and that serum ferritin is secreted by cells rather than being the product of a cytosolic leak from damaged cells. Mouse serum ferritin is composed mostly of L-subunits, whereas it contains few H-subunits and iron content is low. L-subunits of serum ferritin are frequently truncated at the C-terminus, giving rise to a characteristic 17-kD band that has been previously observed in lysosomal ferritin. Taken together with the fact that mouse serum ferritin is not detectably glycosylated, we propose that mouse serum ferritin is secreted through the nonclassical lysosomal secretory pathway.

Hepicidin and its role in iron metabolism

Among various micro-nutrients, iron plays a major role not only for hemoglobin alone but also for oxidative metabolism and energy production. Hemoglobin functions as a chief oxygen carrier while other iron containing mitochondrial enzymes and respiratory chain proteins are involved in oxidative metabolism and release of energy from carbohydrates and fats. Iron salts have very low bioavailability and have potential for toxicity. Therefore iron absorption and its metabolism in humans is highly regulated process. Its regulation at cellular levels is through iron responsive proteins via ironresponsive elements in messenger RNA from genes encoding proteins of iron metabolism. Regulation of iron metabolism along with iron mobilization from macrophages, tissue stores and synthesis of transferrin receptor and ferritin is a complex process. It has been observed that hepatocytes produce hormone peptide hepcidin which plays a major role in the systemic regulation of iron metabolism by its action on target cells at various sites including the liver. Zhang and Enns have recently concluded that hepicidin plays a key of role in the regulation of iron metabolism.1 Several experimental and other clinical studies have demonstrated that hepicidin controls the release of iron from variety of cells such as macrophages, hepatocytes, enterocytes etc to plasma.2-5 Hepcidin primarily controls the iron absorption. However, the recycling of iron from red cell lysis and release of iron from tissue iron stores is carried by the interaction of hepicidin with ferroportin which is a cellular iron exporter in vertebrates.5 Ferroportin is primarily expressed by enterocytes and macrophages. It has been shown that release of ferroportin is also controlled by hepicidin.

Evaluation of serum hepcidin in assessment of iron status in renal transplant patients

Background: Hepcidin is a 25 aminoacid peptide that isproduced primarily by the liver in response to a varietyof stimuli known to modulate tissue iron stores and itsserum availability. It circulates in plasma, is filtered bythe kidney, and accumulates in urine. The presence ofhepcidin in the plasma negatively regulates the egress ofiron from the cells and macrophages, involved intransport of iron into the extracellular spaces. Hepcidinbinds to ferroportin 1 present on the cell surface leadingto the internalization of ferroportin 1 and subsequentdegradation. Hepcidin has also been shown to be anacute phase reactant increased by interleukin 6 (IL-6) andmarkedly induced by infection and inflammation.Documentation of the role of hepcidin in thedevelopment of anemia in chronic kidney disease andwhether it can be used as a marker of iron status inchronic kidney disease (CKD) has been sought in manystudies.The aim of this study was to detect the value of hepcidinas a marker of iron status in post renal transplantrecipients as compared with chronic renal failure (CRF)patients on hemodialysis. Results: In our study we found the level of serumprohepcidin in the renal transplant recipient group to becomparable to the control group. In the group with CKDthe serum prohepcidin was significantly lower thancontrols as well as the transplant recipients (p-value:0.00). There was no significant difference between theCKD group with higher hemoglobin and those withlower hemoglobin. From the correlation between theacute phase proteins and prohepcidin we found a positivecorrelation between serum ferritin, CRP and prohepcidinonly in the group with CKD on hemodialysis. Conclusion: We suggest that serum prohepcidin can be used as a routine measurement in transplant recipients as an indicator of the iron stores in the body. Further dedicated studies on large groups of patients need to be done to verify the usefulness of such a test.

Erythropoietin Injections Stimulate Spleen BMP4-Dependant Stress Erythropoiesis in a Mouse Model of Generalized Inflammation.

Abstract 1983Poster Board I-1005 Introduction:In mouse, acute anemia leads to the rapid expansion and differentiation of stress erythroid progenitors in the spleen. It has been shown that these progenitors respond to BMP4, Stem Cell Factor and hypoxia and differentiate into stress BFU-E. These are sensitive to high levels of erythropoietin (EPO) and rapidly expand in the spleen, allowing rapid recovery from the anemia (JM Perry et al., Blood 2009, 113:911-918). Inflammation is known to inhibit growth and differentiation of erythroid progenitors and to suppress EPO synthesis in the kidney. However, the effect of pro-inflammatory cytokines on this stress erythropoiesis response is not known. We have recently developed a mouse model of zymosan-induced generalized inflammation and shown that stimulation of erythropoiesis by repeated blood withdrawal or injections of erythropoietin favours iron mobilization from tissue iron stores (S. Lasocki et al., CCM 2008, 36:2388-2394), suggesting that EPO treatment may be beneficial provided effective erythropoiesis can be elicited. Objectives:The aim of our study was to assess the impact of EPO injections on the stress erythropoietic response in this mouse model of chronic inflammation. Methods:Mice (C57BL/6) received a single intraperitoneal injection of zymosan at day 1 (Z1) followed by four consecutive daily injections of EPO at day 5, 6, 7 and 8. Mice were analyzed one day (Z9EPO1), four days (Z12EPO4) or nine days (Z17EPO9) after the final injection and compared to controls, Z alone or EPO alone. Double Ter119/CD71 labelling was used to analyze the different stages of erythroblast differentiation by FACS, in bone marrow and spleen in the different conditions. Spleen BMP4 expression was followed by RT-qPCR and immunohistochemistry. Serum EPO levels were measured by ELISA and haematological parameters were recorded. Results:In the inflammatory condition, bone marrow erythropoiesis is suppressed and does not respond to EPO injections. There is a concomitant increased in the percentage of apoptotic Ter119+ cells. In the spleen, inflammation increases spleen size but only moderately stimulates the percentage of erythroblasts. However, EPO injections lead to a 10-fold increase in the percentage of immature erythroblasts at Z9EPO1, followed three days later (Z12EPO4) by a similar increase in the proportion of mature erythroblasts. This finally results in increased reticulocytes and haemoglobin concentration.In the spleen, BMP4 mRNAs are not stimulated by inflammation but significantly increased by EPO injections, both in normal mice and mice with Zymosan-induced inflammation. The protein BMP4 is expressed by erythroid precursors and stromal cells. Double labelling with F4/80 and BMP4 clearly shows that spleen macrophages are the BMP4-expressing cells following EPO injections in mice with a generalized inflammation. Conclusion:In mouse, bone marrow erythropoiesis is repressed by inflammation as it has been shown for human erythropoiesis and it does not respond to EPO injections. By contrast, spleen stress erythropoiesis is strongly stimulated by injections of EPO despite the presence of inflammation. This results from a strong increase in BMP4 synthesis by spleen macrophages. BMP4 is known to be stimulated by acute anemia but our study is the first report of a direct effect of EPO injections on BMP4 expression in the spleen and of the identification of macrophages as the stromal cells producing BMP4.It will be of interest to find out if bone marrow macrophages in humans can synthesize BMP4 and also contribute to a medullar stress erythropoietic response. Disclosures:No relevant conflicts of interest to declare.