Oxidative Stress and Genomic Damage Induced In Vitro in Human Peripheral Blood by Two Preventive Treatments of Iron Deficiency Anemia

Abstract Abstract 3210 Introduction: Iron deficiency anemia (IDA) is a major health problem worldwide. Although a clinical diagnosis is relatively simple, specific laboratory markers of IDA are lacking especially in the setting of inflammation. Ferritin, the current standard to define IDA is an acute phase protein that is non-specifically elevated during inflammation. Serum transferrin receptor level measurements although available are not yet standardized as a clinical tool. Serum hepcidin is a recently developed novel marker that is currently neither available nor standardized sufficiently. Furthermore, such assays require instrumentation, technical sophistication, and are expensive. Objectives: We sought to identify novel markers of IDA using mass spectrometry based proteomics. Identifying such markers could yield targets that once validated could serve cost effective point of care assays to detect iron deficiency anemia. Since there is evidence for oxidative damage mediated by reactive oxygen species in IDA, as a first step, we characterized and quantified posttranslational oxidative modifications of hemoglobin and tested their utility as biomarkers. Patients and Methods: We prospectively enrolled patients with IDA (defined as ferritin <12ng/ml in the presence of normal CRP and/or a bone marrow aspirate with “0” iron stores) and healthy controls (n = 23 and 15 respectively). Patients with diabetes, cardiovascular disease, renal disease, cerebrovascular disease and liver disease were excluded as these are conditions associated with preexisting oxidant stress. Erythrocytes from the blood of IDA patients and controls were isolated by centrifugation, washed in 0.9% saline, and lysed in distilled water to yield intracellular hemoglobin. Hemoglobin was then either studied further as an intact molecule or after digestion with trypsin. We used matrix assisted laser desorption ionization (MALDI - TOF) mass spectrometry to identify oxidative modifications of tryptic digested hemoglobin. We used electro spray ionization (ESI) mass spectrometry to identify and semiquantitate oxidative hemoglobin modifications by methods previously established and published by others and ourselves. Results: Using a combination of mass spectrometric methods, we identified 4 oxidative modifications of hemoglobin in patients with IDA and healthy controls (Table 1). Interestingly, a non enzymatic posttranslational modification of hemoglobin, glutathionyl hemoglobin, was found to be significantly increased in IDA patients compared with healthy controls (Glutathionyl hemoglobin % of beta chain; mean ± SD 0.169 ± 0.096 vs 0.077 ± 0.037; p = 0.001). Markers of oxidative stress (reduced RBC glutathione) were lower in IDA compared to healthy controls but the difference was not significant (mean ± SD 0.92 ± 0.53 vs 1.08 ± 0.52 mmol/L; p = 0.54). Glutathionyl hemoglobin levels correlated inversely with serum ferritin (Spearman rho -0.485; p < 0.05). Conclusions: Using two distinct proteomic methods, we identified oxidative posttranslational modifications of hemoglobin in IDA and healthy controls. Glutathionyl hemoglobin, an established marker of oxidative stress was elevated in patients with IDA and correlated inversely with serum ferritin. Overall, these findings suggest that glutathionyl hemoglobin has potential as a biomarker of IDA. Disclosures: No relevant conflicts of interest to declare.

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Is the Peripheral Blood Film Reliable for the Diagnosis of Iron Deficiency Anemia?

American Journal of Clinical Pathology ◽

10.1093/ajcp/55.4.447 ◽

1971 ◽

Vol 55 (4) ◽

pp. 447-451 ◽

Cited By ~ 22

Author(s):

Virgil F. Fairbanks

Keyword(s):

Iron Deficiency ◽

Iron Deficiency Anemia ◽

Peripheral Blood ◽

Blood Film ◽

Deficiency Anemia ◽

Peripheral Blood Film

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Long-term Effect of Intravenous Iron Carboxymaltose Treatment on Oxidative Stress in Women with Iron Deficiency Anemia

Volume 1, Issue 3 - Health Sciences Quarterly ◽

10.26900/hsq.1.1.02 ◽

2021 ◽

pp. 3-10

Author(s):

Gökhan Pektaş ◽

İsmail Kırlı

Keyword(s):

Oxidative Stress ◽

Nitric Oxide ◽

Iron Deficiency ◽

Iron Deficiency Anemia ◽

Catalase Activity ◽

Intravenous Iron ◽

Deficiency Anemia ◽

Healthy Controls ◽

Iron Treatment ◽

Oxidant Status

Objective: This study aims to clarify the effects of intravenous iron supplementation on biomarkers for oxidative stress in women with iron deficiency anemia. Methods: This is a cross-sectional review of 40 healthy women and 40 women who underwent intravenous iron treatment due to anemia. Biochemical markers for oxidative stress were determined for both healthy controls and anemic patients. These markers were also evaluated at hour 1 and day 30 of intravenous iron treatment. Results: The patients with anemia had significantly higher catalase activity and total oxidant status (TOS) but significantly lower nitrate and total anti-oxidant status (TAS) than the healthy controls (p=0.0245, p<0.0001, p=0.0437 and p<0.0001 respectively). At hour 1 of intravenous iron treatment, nitrate, nitrite, nitric oxide, total thiol and TAS values were significantly lower and TOS values were significantly higher than those before the administration of treatment (p=0.0322, p=0.0003, p=0.0005, p<0.0001, p<0.0001 and p=0.004). At day 30 of intravenous iron treatment, catalase activity, nitrate, total thiol and TOS values were significantly lower than those before the administration of treatment (p=0.0332, p=0.0015, p=0.0391 and p<0.0001 respectively) and at hour 1 of treatment (p=0.0498, p<0.0001, p=0.0004 and p<0.0001 respectively). At day 30 of intravenous iron treatment, nitric oxide and TAS values were significantly higher than those before the administration of treatment (p=0.0480 and p=0.001 respectively) and at hour 1 of treatment (p<0.0001 for both). Conclusion: Intravenous iron replacement prompts oxidative stress at hour 1 of infusion in adults with anemia but this increase resolves partially in the following 30 days.

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Clinical Characteristics of Patients with a RBC Fragment Flag on the Advia® 2120 Automated Hematology Analyzer.

Blood ◽

10.1182/blood.v110.11.5153.5153 ◽

2007 ◽

Vol 110 (11) ◽

pp. 5153-5153

Author(s):

Jonathan Ben-Ezra

Keyword(s):

Iron Deficiency ◽

Iron Deficiency Anemia ◽

Peripheral Blood ◽

Blood Smear ◽

Lymphoblastic Leukemia ◽

Peripheral Blood Smear ◽

Deficiency Anemia ◽

Clinical Criteria ◽

Hematology Analyzer ◽

Automated Hematology Analyzer

The clinical diagnosis of thrombotic thrombocytopenia purpura (TTP) is a difficult one to make. It is based on clinical criteria, one of which is a microangiopathic hemolytic anemia, characterized morphologically by the presence of schistocytes on the peripheral blood smear. The ADVIA 2120 automated hematology analyzer is able to quantify the presence of red blood cell (RBC) fragments. We studied the ability of the ADVIA 2120 to be able to detect RBC fragments in the blood of TTP patients, and the characteristics of all patients in whom RBC fragments were obtained. During the study period, 6 TTP patients were studied. The initial numbers of RBC fragments ranged from 0.02–0.05 × 106 cells/μl. During the course of plasmapheresis, these numbers decreased to 0.00–0.02 × 106 cells/μl, corresponding to a rise in the platelet count. Figure Figure In the course of a month, 52 blood samples on 39 patients were flagged by the hematology analyzer to have RBC fragments (0.01–0.12 × 106 cells/μl). 52 Samples with RBC Fragment Flag Hemoglobin Platelets RDW Range 4– 14.3 g/dl 5–906 × 103/ul 13.9– 28.6% Number Abnormal 46 (<13.0 g/dl) 23 (<160 × 103/ul) 51 (>14.1%) Within this population, there were two patients with TTP, and one with DIC. Four of the samples did not have detectable schistocytes upon visual inspection of the peripheral blood smear. There were 19 samples from 14 patients who had RBC fragment counts ≥ 0.04 × 106 cells/μl. 19 Specimens with RBC Fragments ≥ 0.04 × 106/ul Hemoglobin Platelets RDW Range 8– 14.1 g/dl 59– 906 × 103/ul 16.4– 25.3% Number Abnormal 15 (<13 g/dl) 4 (<160 × 103/ul) 19 (>14.1%) The diagnoses in these 14 patients were iron deficiency anemia (4 patients), thalassemia trait (2), acute lymphoblastic leukemia (2), and one each with TTP, sickle cell anemia, heart failure, kidney stone, cerebrovascular accident (CVA), and end stage renal disease. We conclude that the RBC fragment flag on the ADVIA 2120 is nonspecific. Although it does detect schistocytes in TTP, these are often present in low numbers. Quantitatively, the most numerous RBC fragments are found in diseases with marked anisopoikilocytosis, such as iron deficiency anemia.

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Effect of iron deficiency anemia on lipid peroxidation and antioxidant systems

Journal of College of Medical Sciences-Nepal ◽

10.3126/jcmsn.v7i4.6739 ◽

2012 ◽

Vol 7 (4) ◽

pp. 34-43 ◽

Cited By ~ 1

Author(s):

NL Madhikarmi ◽

KRS Murthy

Keyword(s):

Oxidative Stress ◽

Lipid Peroxidation ◽

Iron Deficiency ◽

Iron Deficiency Anemia ◽

Binding Capacity ◽

Lipid Hydroperoxide ◽

Nutritional Deficiencies ◽

Deficiency Anemia ◽

Antioxidant Systems ◽

Enzymatic Antioxidants

Iron deficiency is one of the most widespread nutritional deficiencies in the world. Globally more than two billion people are suffering from iron deficiency anemia. The present study was designed to investigate the influence of a large number of factors known to be associated with oxidative stress. The case-control study determines the lipid peroxidation and antioxidants status in forty Iron deficiency anemia and forty healthy volunteers with their informed consent. All the parameters were assayed by spectrophotometric methods. Blood hemoglobin and plasma iron were decreased whereas total iron binding capacity was increased significantly in Iron deficiency anemia. The lipid peroxidation parameters like malondialdehyde, lipid hydroperoxide were significantly increased in Iron deficiency anemia. Both enzymatic; glutathione peroxidase, superoxide dismutase and catalase and non-enzymatic antioxidants; vitamin C, E and reduced glutathione were significantly decreased in Iron deficiency anemia case as compared to their healthy counterparts. Our findings suggest, increased lipid peroxidation products and reduced antioxidants system boost the oxidative stress state, hence deteriorating the condition of Iron deficiency anemia patients. Journal of College of Medical Sciences-Nepal,2011,Vol-7,No-4, 34-43 DOI: http://dx.doi.org/10.3126/jcmsn.v7i4.6739

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