Free Iron Content Research Articles

Oxidative stress in pediatric patients with β thalassemia major

Background β-thalassemia major (β-TM) is a common inherited hemolytic type of anemia. Repeated blood transfusions predispose β-TM patients toward peroxidative tissue injury because of secondary iron overload. Objectives This study aimed to evaluate the effects of iron overload on antioxidant enzymes and liver cell damage in β-TM patients undergoing regular blood transfusions. Patients and methods This prospective case–control cohort study included 30 pediatric patients with a confirmed diagnosis of β-TM on regular blood transfusions and 20 age-matched and sex-matched healthy children attending the Qena University Hospital, Pediatric Clinic. Blood samples were withdrawn from each patient to measure serum levels of ferritin, glutathione peroxidase (GPX), and superoxide dismutase (SOD). Results Total bilirubin, aspartate transaminase (AST), alanine transaminase (ALT), and ferritin levels were significantly higher in the β-TM group (P Conclusion Impaired levels of antioxidant enzymes SOD and GPX in patients with β-TM on repeated transfusion, in addition to excessive free iron concentration, iron overload may attribute to oxidative damage in these patients. Antioxidant systems that compensate for reduced lipid peroxidation to lower tissue damage are needed.

THU-406 - Elevated faecal free iron content in HFE haemochromatosis is reduced by therapeutic venesection

THU-406 - Elevated faecal free iron content in HFE haemochromatosis is reduced by therapeutic venesection

Iron Profile and Glycaemic Control in Patients with Type 2 Diabetes Mellitus

Iron overload is increasingly being connected to insulin resistance in Type 2 Diabetes Mellitus (T2DM) patients. Free iron causes the assembly of reactive oxygen species that invariably steer the body’s homeostasis towards oxidative stress-mediated diabetic complications. This study aims to assess the serum iron, total iron binding capacity (TIBC), and percentage transferrin saturation (Tsat) of 150 subjects divided into three groups (I,II,III) of 50. Healthy individuals (controls) constituted Group I. Group II consisted of T2DM patients with optimal glycaemic control. T2DM patients with suboptimal glycaemic control formed group III. Mean serum free iron concentration was 105.34 ± 3.5, 107.33 ± 3.45, and 125.58 ± 3.45 μg/dL in Group I, Group II, and Group III, respectively. Mean serum TIBC concentration in Group I, Group II, and Group III was 311.39 ± 5.47, 309.63 ± 6.1, and 284.2 ± 3.18μg/dL, respectively. Mean serum transferrin saturation (%) in Group I, Group II, and Group III was 34.17 ± 1.21, 35.02 ± 1.2, and 44.39 ± 1.07, respectively. The difference between TIBC, mean serum free iron concentration, and transferrin saturation between Group I and Group III (for all, p values <0.001), as well as between Group II and Group III (p values 0.0012, 0.0015, and <0.0001, respectively) was statistically significant. The fasting plasma glucose values of Groups II and III were significantly higher than those of Group I, (p < 0.0001). Glycated haemoglobin (HbA1c) values were also shown to increase from Group I to II and then III, and the increase was highly significant (all p values <0.0001). Thus, decreased glycaemic control and an increase in the glycation of haemoglobin was the key to elevation in serum iron values and alterations in other parameters. However, a significant correlation was absent between serum iron and HbA1c (r = 0.05) and transferrin saturation (r = 0.0496) in Group III.

Iron-induced nitric oxide leads to an increase in the expression of ferritin during the senescence of Lotus japonicus nodules

Iron is an essential nutrient for legume-rhizobium symbiosis and accumulates abundantly in the nodules. However, the concentration of free iron in the cells is strictly controlled to avoid toxicity. It is known that ferritin accumulates in the cells as an iron storage protein. During nodule senescence, the expression of the ferritin gene, Ljfer1, was induced in Lotus japonicus. We investigated a signal transduction pathway leading to the increase of Ljfer1 in the nodule. The Ljfer1 promoter of L. japonicus contains a conserved Iron-Dependent Regulatory Sequence (IDRS). The expression of Ljfer1 was induced by the application of iron or sodium nitroprusside, which is a nitric oxide (NO) donor. The application of iron to the nodule increased the level of NO. These data strongly suggest that iron-induced NO leads to increased expression of Ljfer1 during the senescence of L. japonicus nodules.

Effect of grape seed and skin supplement on milk yield and composition of dairy ewes.

In the present study, we investigated the effect of grape seed and skin supplement (GSSS), on lactating dairy ewes' production. Ten dairy pregnant ewes from northern Tunisia were allocated to two groups: control diet (C) and supplemented with 20% (w/w) GSSS. The experiment lasted for 8weeks and took place after 2months of lambing. During the experiment, daily milk yield and milk composition were determined. Supplementation of the diet with GSSS increased milk production (P < 0.001), calcium (P < 0.01), free iron (P < 0.01) and urea content (P < 0.001) but had no effect on milk fat nor protein. From these data, it is concluded that the inclusion of GSSS in sheep diets increased significantly ewes' milk yield.

Structural basis of haem-iron acquisition by fungal pathogens

Pathogenic microorganisms must cope with extremely low free-iron concentrations in the host's tissues. Some fungal pathogens rely on secreted haemophores that belong to the Common in Fungal Extracellular Membrane (CFEM) protein family, to extract haem from haemoglobin and to transfer it to the cell's interior, where it can serve as a source of iron. Here we report the first three-dimensional structure of a CFEM protein, the haemophore Csa2 secreted by Candida albicans. The CFEM domain adopts a novel helical-basket fold that consists of six α-helices, and is uniquely stabilized by four disulfide bonds formed by its eight signature cysteines. The planar haem molecule is bound between a flat hydrophobic platform located on top of the helical basket and a peripheral N-terminal 'handle' extension. Exceptionally, an aspartic residue serves as the CFEM axial ligand, and so confers coordination of Fe3+ haem, but not of Fe2+ haem. Histidine substitution mutants of this conserved Asp acquired Fe2+ haem binding and retained the capacity to extract haem from haemoglobin. However, His-substituted CFEM proteins were not functional in vivo and showed disturbed haem exchange in vitro, which suggests a role for the oxidation-state-specific Asp coordination in haem acquisition by CFEM proteins.

Electrochemistry of Tris(1,10-phenanthroline)iron(II) inside a polymeric hydrogel. Coupled chemical reactions and migration effects

The retention, release, and detection of metallic complexes in polymeric hydrogels are of interest in drug delivery, analytical chemistry, and water remediation. The electrochemistry of the redox complexes inside the hydrogel could be affected by the viscoelastic properties of the gel, local ionic force and pH, and interactions (e.g., hydrophobic) between the complex and the polymer chains. In this work, it is shown that a simple setup, consisting of a disk electrode pressed on the hydrogel, allows to perform electrochemistry of a redox couple: Tris(1,10-phenanthroline)iron(II) (Fe(phen)3 2+) inside a hydrogel matrix. The behavior is compared with the same couple in solution, and it is found that the electrochemical properties of the redox couple are strongly affected by the presence of the hydrogel matrix. The cyclic voltammogram of the hydrogel loaded with complex shows a response, which suggests electrochemical-chemical mechanism. The chemical step is likely linked to a catalytic oxidation of free hydrated Fe2+ ions present inside the hydrogel together with the redox complex. Since Fe2+ ions have small charge transfer constants on the glassy carbon electrodes, only the catalytic current is observed. Indeed, when excess ligand (phenanthroline) is absorbed inside the hydrogel, the measured cyclic voltammograms show a single reversible oxidation/reduction step. It seems that the complexation equilibrium shifts toward the complex, making the free iron concentration negligible. Accordingly, the cyclic voltammetry shape and peak potential difference agree with a reversible oxidation/reduction. Additionally, the peak currents of the cyclic voltammograms show a linear dependence with the square root of time, as predicted by a Randles-Sevcik equation. However, the measured currents are smaller than the simulated ones. The differences are in agreement with simulations of the cyclic voltammograms where the migration of the redox species is considered. Chronoamperometry is used to measure the mass transport of redox species inside the hydrogel. It is found that the current transients still obey Cottrell’s equation, but the diffusion coefficients obtained from the slopes of Cottrell’s plots have to be corrected for migration effects. The effective diffusion coefficient of Fe(phen)3 2+ measured inside the hydrogel (D Red-hydrogel = 5.5 (±0.5) × 10−8 cm2 s−1) is ca. 80 times smaller than the one measured in solution (D Red-solution = 4.4 (±0.5) × 10−6 cm2 s−1). The simple setup has a true semi-infinite boundary condition, which allows characterizing the hydrogel in the same condition as the bulk material and easily changing both the redox species and the hydrogel structure.

A Streptococcus suis LysM domain surface protein contributes to bacterial virulence

Streptococcus suis (SS) is a major swine pathogen, as well as a zoonotic agent for humans. Numerous factors contribute to SS virulence, but the pathogenesis of SS infection is poorly understood. Here, we show that a novel SS surface protein containing a LysM at the N-terminus (SS9-LysM) contributes to SS virulence. Homology analysis revealed that the amino acid sequence of SS9-LysM from the SS strain GZ0565 shares 99.8–68.7% identity with homologous proteins from other SS strains and 41.2% identity with Group B Streptococcal protective antigen Sip. Immunization experiments showed that 7 out of 30 mice immunized with recombinant SS9-LysM were protected against challenge with the virulent GZ0565 strain, while all of the control mice died within 48h following bacterial challenge. In mouse infection model, the virulence of the SS9-LysM deletion mutant (ΔSS9-LysM) was reduced compared with the wild-type (WT) strain GZ0565 and SS9-LysM complemented strain. In addition, ΔSS9-LysM was significantly more sensitive to killing by pig blood ex vivo and mouse blood in vivo compared with the WT strain and SS9-LysM complemented strain. In vivo transcriptome analysis in mouse blood showed that the WT strain reduced the expression of host genes related to iron-binding by SS9-LysM. Moreover, the total free iron concentration in blood from infected mice was significantly lower for the ΔSS9-LysM strain compared with the WT strain. Together, our data reveal that SS9-LysM facilitates SS survival within blood by releasing more free iron from the host. This represents a new mechanism of SS pathogenesis.

Correlation between Antioxidant Enzyme Activity, Free Iron Content and Lipid Oxidation in Four Lines of Korean Native Chicken Meat.

This study was conducted to observe the association between antioxidant enzyme activity, free iron content and lipid oxidation of Korean native chicken (KNC) meat during refrigerated storage. Four lines of KNC (Yeonsan ogye, Hyunin black, Hoengseong yakdak and Hwangbong) were raised under similar conditions. A total of 16 roosters were randomly sampled and slaughtered at the age of 12 mon. The breast and thigh meats were stored aerobically for 10 d at 4℃. Although thigh meat had higher antioxidant enzyme activity, it was more susceptible to lipid oxidation and released more iron during storage than breast meat. Aerobic refrigerated storage for 10 d significantly decreased the activity of antioxidant enzymes and increased the amount of free iron and malondialdehyde. The activities of superoxide dismutase (SOD) and glutathione peroxidase (GSH-Px) were negatively correlated with lipid oxidation, whereas that of catalase was not. The amount of free iron was positively associated with lipid oxidation. We concluded that chicken line did not affect strongly on antioxidant enzyme activity and lipid oxidation in breast meat of KNC. However, the thigh meat of Hwangbong and Hyunin black had higher SOD and GSH-Px activity, respectively, and lower malondialdehyde contents than that of other chickens. SOD, GSH-Px and free iron play significant roles in meat lipid oxidation during refrigerated storage.

HOMOLOGY MODELING AND TOPOLOGY PREDICTION OF FRPBPROTEININ NEISSERIA MENINGITIDIS

Iron is an essential nutrient for almost all bacteria, but the concentration of free iron in the human host is too low to support growth. In the extracellular compartments of the human body, iron is bound mainly to transferrin and lactoferrin. Many bacteria deal with iron limitation by the secretion of siderophores, which are small organic compounds that bind iron very efficiently. The Fe31-siderophore complex is bound to specific receptors on the bacterial cell surface and internalized. The transport across the outer membrane requires energy, which is provided in Escherichia coli by the inner membrane protein TonB.FrpB is another iron-limitation inducible outer membrane protein of Neisseria meningitides. Its function is unknown. Previously, we have raised monoclonal antibodies against this protein, most of which appeared to be bactericidal in an in vitro assay with human complement. Here we describe the topology and 3D structure of a novel antigen which was discovered by mining the bacterial genome and that is very effective in inducing bactericidal antibodies. This antigen is a very good candidate for inclusion in universal vaccines against N.meningitidis.

Association of serum free iron and glycemic control among type II diabetes mellitus population in Puducherry - a preliminary study

Background: Iron modify glucose metabolism and glucose metabolism impinges on iron. Glycation of hemoglobin contributes to substantial affinity for transitional metals and the development of complications is linked to the accumulation of glycation. Objective: This study was undertaken to assess the serum free iron levels in patients with type II diabetes mellitus (DM) and its correlation with good and poor glycemic control. Materials and Methods: An analytical cross-sectional study was conducted among 100 patients and 50 healthy controls. They are grouped as Group I, 50 normal healthy controls; Group II, 50 type II DM patients with good glycemic control (HbA1C ≤ 7); Group III, 50 type II DM patients with poor glycemic control (HbA1C > 7). Levels of HbA1C l and serum iron were assessed. The data collected were recorded and analyzed using SPSS version 16.0. Result: The mean serum iron concentration between group I and III (p < 0.001) as well as between group II and III (p < 0.005) was statistically significant. Difference between group I and II was not statistically significant (p = 0.205). Conclusion: From our study it was concluded that serum free iron concentration was higher in patients with type II DM with poor glycemic control. In addition, there was a positive correlation between serum free iron concentration and glycemic control.

Mechanism of Iron-Dependent Repressor (IdeR) Activation and DNA Binding: A Molecular Dynamics and Protein Structure Network Study.

Metalloproteins form a major class of enzymes in the living system that are involved in crucial biological functions such as catalysis, redox reactions and as ‘switches’ in signal transductions. Iron dependent repressor (IdeR) is a metal-sensing transcription factor that regulates free iron concentration in Mycobacterium tuberculosis. IdeR is also known to promote bacterial virulence, making it an important target in the field of therapeutics. Mechanistic details of how iron ions modulate IdeR such that it dimerizes and binds to DNA is not understood clearly. In this study, we have performed molecular dynamic simulations and integrated it with protein structure networks to study the influence of iron on IdeR structure and function. A significant structural variation between the metallated and the non-metallated system is observed. Our simulations clearly indicate the importance of iron in stabilizing its monomeric subunit, which in turn promotes dimerization. However, the most striking results are obtained from the simulations of IdeR-DNA complex in the absence of metals, where at the end of 100ns simulations, the protein subunits are seen to rapidly dissociate away from the DNA, thereby forming an excellent resource to investigate the mechanism of DNA binding. We have also investigated the role of iron as an allosteric regulator of IdeR that positively induces IdeR-DNA complex formation. Based on this study, a mechanistic model of IdeR activation and DNA binding has been proposed.

Iron metabolism following intravenous transfusion with stored versus fresh autologous erythrocyte concentrate in healthy dogs.

OBJECTIVE To determine effects of IV transfusion with fresh (3-day-old) or stored (35-day-old) autologous erythrocyte concentrate on serum labile iron concentration, iron-binding capacity, and protein interaction with iron in dogs. ANIMALS 10 random-source healthy dogs. PROCEDURES Dogs were randomly assigned to receive autologous erythrocyte concentrate stored for 3 days (n = 5) or 35 days (5). One unit of whole blood was collected from each dog, and erythrocyte concentrates were prepared and stored as assigned. After erythrocyte storage, IV transfusion was performed, with dogs receiving their own erythrocyte concentrate. Blood samples were collected from each dog before and 5, 9, 24, 48, and 72 hours after transfusion. Serum was harvested for measurement of total iron, labile iron, transferrin, ferritin, hemoglobin, and haptoglobin concentrations. RESULTS For dogs that received fresh erythrocytes, serum concentrations of the various analytes largely remained unchanged after transfusion. For dogs that received stored erythrocytes, serum concentrations of total iron, labile iron, hemoglobin, and ferritin increased markedly and serum concentrations of transferrin and haptoglobin decreased after transfusion. CONCLUSIONS AND CLINICAL RELEVANCE Transfusion with autologous erythrocyte concentrate stored for 35 days resulted in evidence of intravascular hemolysis in healthy dogs. The associated marked increases in circulating concentrations of free iron and hemoglobin have the potential to adversely affect transfusion recipients.

Cationic Peptides Facilitate Iron-induced Mutagenesis in Bacteria

Pseudomonas aeruginosa is the causative agent of chronic respiratory infections and is an important pathogen of cystic fibrosis patients. Adaptive mutations play an essential role for antimicrobial resistance and persistence. The factors that contribute to bacterial mutagenesis in this environment are not clear. Recently it has been proposed that cationic antimicrobial peptides such as LL-37 could act as mutagens in P. aeruginosa. Here we provide experimental evidence that mutagenesis is the product of a joint action of LL-37 and free iron. By estimating mutation rate, mutant frequencies and assessing mutational spectra in P. aeruginosa treated either with LL-37, iron or a combination of both we demonstrate that mutation rate and mutant frequency were increased only when free iron and LL-37 were present simultaneously. Colistin had the same effect. The addition of an iron chelator completely abolished this mutagenic effect, suggesting that LL-37 enables iron to enter the cells resulting in DNA damage by Fenton reactions. This was also supported by the observation that the mutational spectrum of the bacteria under LL-37-iron regime showed one of the characteristic Fenton reaction fingerprints: C to T transitions. Free iron concentration in nature and within hosts is kept at a very low level, but the situation in infected lungs of cystic fibrosis patients is different. Intermittent bleeding and damage to the epithelial cells in lungs may contribute to the release of free iron that in turn leads to generation of reactive oxygen species and deterioration of the respiratory tract, making it more susceptible to the infection.

The effect of ferrous-chelating hairtail peptides on iron deficiency and intestinal flora in rats.

Chelating agents, such as small peptides, can decrease free iron content and increase iron bioavailability. They may have promising therapeutic potential and may prevent the pro-oxidant effects of low molecular weight iron. Hairtail is a species of fish that is rich in easily digestible proteins. We extended this strategy for iron delivery by using an enzymatic hydrolysate of hairtail as the chelating agent and found that the ferrous-chelating hairtail peptides have anti-anaemic activity in Sprague-Dawley rats with anaemia. The anti-anaemic activity of ferrous-chelating hairtail peptides prepared by enzymatic hydrolysis of the hairtail and ferrous chelation was studied in rat models of iron deficiency anaemia. After the end of the 35 d experiment, we noted significant differences in haemoglobin, mean corpuscular volume, haemoglobin distribution width, and ferritin concentrations between those animals supplemented with ferrous-chelating hairtail peptides and FeSO4 and healthy animals. There were no negative side effects on the animals' growth or behaviour. There was no obvious inflammation in the intestinal mucosa lamina propria and no unbalance of intestinal flora. The novel ferrous-chelating hairtail peptides may be a suitable fortificant for improving iron-deficiency status. Our findings demonstrated that this multi-tracer technique has many applications in nutritional research. © 2015 Society of Chemical Industry.

Effect of humic substances and clay minerals on the hydrosorption capacity of clay particles in meadow soils (Middle Priamurje, Far East of Russia)

The study of clay fractions is usually focused on water-dispersible clay (WDC), which is a good indicator of many soil processes involving water (water erosion, transfer and removal of nutrients and contaminants). Aggregated clay (AC) that remains after WDC extraction is also important, as it controls the formation of aggregates and their mechanical and water resistance. The aim of this study was a gradual extraction of WDC and AC fractions from periodically overmoistening meadow podbel (mollic planosol) and the study of the content and composition of humic substances and clay minerals in these fractions as well as their effect on the hydrosorption capacity. The complete extraction of clay fractions showed that AC prevails over WDC along the entire profile and most evidently in the Ag horizon. Most of the organic matter is concentrated in AC. X-ray diffractometry and SEM analysis revealed a similar set of minerals in both fractions, but in different proportions. In the Ag and Elcg horizons, WDC is enriched with clastic components (fine-dispersed quartz, feldspars, amphiboles) and clay minerals with rigid structures (illite, kaolinite, chlorite), compared to AC. In the Btg1 horizon smectite prevails in both fractions. The specific surface, which characterizes the hydrosorption capacity of clays, of WDC is 28–95 m2/g less than that of AC. Maximal difference in specific surface of clays is found in the horizon Elcg, where AC has the highest content of free iron. The present study allowed defining the heterogeneity of the total clay in the periodically overmoistened meadow soil, which is manifested in differences in composition and properties of WDC and AC fractions. AC largely determines hydrosorption properties of soil due to a high content of organic matter and the predominance of swelling minerals in its composition, compared with WDC. The obtained data highlight the importance of studying clay fractions WDC and AC for the characteristics of differentiation and aggregation of the clay in soils.

Chemistry of Arsenic in Semi-Arid Alkaline Soils of the Southern High Plains, USA: Sorption Characteristics and Interactions with Soil Constituents

Limited information is available on the chemistry of arsenic (As) in the semi-arid alkaline soils of the Southern High Plains (SHP), USA. This study examined As sorption characteristics and its interactions with soil constituents in important agricultural soils (Amarillo, Arvana, Patricia, and Pullman) of the SHP using sorption isotherm models and sequential fractionation techniques. Results from fractionation of As into five distinct pools showed that about 52.4 % of the added As was found in the exchangeable and non-adsorbed pool in the Amarillo soil, suggesting this soil could have the highest tendency to release sorbed As to the environment, whereas the Pullman soil exhibited the greatest capacity to fix As as over 45 % of the added As was found in the residual fraction. The distribution of As among the soil constituents was a reflection of the characteristics of these soils. Arsenic sorption behaviors were well described by both the Freundlich and Langmuir models. Arsenic sorption maxima (q max) was highest in the Amarillo soil (~2124 mg kg−1), followed by the Pullman, Arvana, and Patricia soils with q max values of 1692, 1370, and 1317 mg kg−1, respectively. The Freundlich distribution coefficient (K d) was highest in the Pullman soil (21.6 L kg−1) and lowest in the Amarillo (1.38 L kg−1). Sorption parameters such as K d, N (sorption intensity constant) and qmax, varied among the soils, and the variability associated with K d and N in these semi-arid soils was explained by soil properties such as pH, organic matter, calcium carbonate, total free iron (Fe), sand, clay, total aluminum, total Fe, and total manganese contents. Findings from this study are important in understanding the environmental fate of As in semi-arid/arid climates and could be extended to other regions with similar soil characteristics.

Dating Endometriotic Ovarian Cysts Based on the Content of Cyst Fluid and its Potential Clinical Implications.

This study was undertaken to test the hypotheses that, due to gradual accumulation of dead erythrocytes and their ingested products resulting from repeated hemorrhage, older endometriomas (whitish in color) contain chocolate fluid with higher iron content than younger (brownish/blackish in color) ones with concomitant higher collagen content and more adhesions. We recruited 30 premenopausal women with histologically confirmed ovarian endometriomas and collected samples of their endometriotic lesions and chocolate fluid and measured the viscosity, density, and the concentration of total bilirubin, ferritin, and free iron of the chocolate fluid. We also evaluated the lesion color and adhesion scores. In addition, we performed Masson trichrome and Picro-Sirius red staining on all endometriotic cysts and evaluated the extent of fibrosis in the lesions. We found that fluids taken from white-colored endometriomas had significantly higher concentration of total bilirubin, ferritin, and free iron, respectively, than black/brown-colored ones. In addition, older cysts had fluids that had significantly higher density and viscosity. Fluid density correlated positively with the concentrations of total bilirubin, ferritin, and free iron. Older lesions had significantly more collagen content and higher adhesion scores. Taken together, these data supports the notion that older cysts, having experienced more bleeding episodes, contain chocolate fluid that is higher in viscosity, density, and iron content and higher fibrotic content than younger ones. This provides another piece of evidence that endometriotic lesions are wounds that undergo repeated injury and repair, resulting ultimately fibrotic lesions that are resistant to hormonal treatment.

Effects of changes in intracellular iron pool on AlkB-dependent and AlkB-independent mechanisms protecting E.coli cells against mutagenic action of alkylating agent

An Escherichia coli hemH mutant accumulates protoporphyrin IX, causing photosensitivity of cells to visible light. Here, we have shown that intracellular free iron in hemH mutants is double that observed in hemH+ strain. The aim of this study was to recognize the influence of this increased free iron concentration on AlkB-directed repair of alkylated DNA by analyzing survival and argE3→Arg+ reversion induction after λ>320nm light irradiation and MMS-treatment in E. coli AB1157 hemH and alkB mutants. E.coli AlkB dioxygenase constitutes a direct single-protein repair system using non-hem Fe(II) and cofactors 2-oxoglutarate (2OG) and oxygen (O2) to initiate oxidative dealkylation of DNA/RNA bases. We have established that the frequency of MMS-induced Arg+ revertants in AB1157 alkB+hemH–/pMW1 strain was 40 and 26% reduced comparing to the alkB+hemH– and alkB+hemH+/pMW1, respectively. It is noteworthy that the effect was observed only when bacteria were irradiated with λ>320nm light prior MMS-treatment. This finding indicates efficient repair of alkylated DNA in photosensibilized cells in the presence of higher free iron pool and AlkB concentrations. Interestingly, a 31% decrease in the level of Arg+ reversion was observed in irradiated and MMS-treated hemH–alkB– cells comparing to the hemH+alkB– strain. Also, the level of Arg+ revertants in the irradiated and MMS treated hemH– alkB– mutant was significantly lower (by 34%) in comparison to the same strain but MMS-treated only. These indicate AlkB-independent repair involving Fe ions and reactive oxygen species. According to our hypothesis it may be caused by non-enzymatic dealkylation of alkylated dNTPs in E. coli cells. In in vitro studies, the absence of AlkB protein in the presence of iron ions allowed etheno(ϵ) dATP and ϵdCTP to spontaneously convert to dAMP and dCMP, respectively. Thus, hemH– intra-cellular conditions may favor Fe-dependent dealkylation of modified dNTPs.

MtDNA mutagenesis impairs elimination of mitochondria during erythroid maturation leading to enhanced erythrocyte destruction.

Haematopoietic progenitor cells show special sensitivity to mitochondrial DNA (mtDNA) mutagenesis, which suggests that increased mtDNA mutagenesis could underlie anemias. Here we show that elevated mtDNA mutagenesis in mice with a proof-reading deficient mtDNA polymerase (PolG) leads to incomplete mitochondrial clearance, with asynchronized iron loading in erythroid precursors, and increased total and free cellular iron content. The resulting Fenton chemistry leads to oxidative damage and premature destruction of erythrocytes by splenic macrophages. Our data indicate that mitochondria actively contribute to their own elimination in reticulocytes and modulate iron loading. Asynchrony of this sequence of events causes severe mitochondrial anaemia by depleting the organism of red blood cells and the bone marrow of iron. Our findings account for the anaemia development in a progeroid mouse model and may have direct relevance to the anemias associated with human mitochondrial disease and ageing.