Effect of iron supplementation on oxidative stress and antioxidant status in iron-deficiency anemia

This study was designed to measure the effect of iron supplementation on antioxidant status in iron-deficient anemia, including the time for hemoglobin normalization and at the time of filling of iron body stores. The extent of plasma lipid peroxidation was evaluated by measuring the levels of malondialdehyde and glutathione peroxidase (GSH-Px), and the activities of superoxide dismutase (SOD) and catalase in 63 patients with iron-deficiency anemia before and after 6 wk of iron supplementation and at the time when body iron stores are saturated. After 6 wk of iron supplementation, a significant decrease of oxidative stress was observed in the treated subjects relative to controls (p<0.05). No significant differences existed between treated patients at 6 wk and at the end of the study. The erythrocyte levels of catalase, SOD, and GSH-Px were significantly lower in treated patients relative to controls (p<0.05). These levels increased after 6 wk of supplementation (p<0.05) and showed no significant differences with those at the end of the study.     

In vitro and in vivo effects of selenium and selenium with vitamin E on platelet functions in diabetic rats relationship to platelet sorbitol and fatty acid distribution

In vitro 30 min of incubation with selenomethionine (Sm)+vitamin E multiplied by about five platelet selenium (Se) decreased significantly platelet thrombin and ADP-induced aggregation decrease. Four groups of streptozotocin-induced diabetic rats were fed with a supplemented purified diet with an Se-rich yeast (Selenion): DSel, Sm: DSm, Sm α-tocopherol: DSmE or unsupplemented diet: D. After 24 wk of supplementation, only a decrease in thrombin-induced aggregation in group DSel compared to DSm and DSmE and D was observed. However, after 24 wk of diet compared to 14 wk, in group D and DSm, a significant increase in thrombin-induced aggregation occurred (p<0.0001), whereas a significant decrease in groups DSel and DSmE (p<0.0001,p<0.03) was noted. After 21 wk of diet, in DSmE, platelet adhesion to fibronectin was significantly decreased compared to group D (p<0.05). These changes in DSmE were associated with a significant decrease in platelet sorbitol (p<0.02) and a very significant increase in platelet Se (p<0.0005). Sm associated with vitamin E would appear more efficient to prevent oxidative damage of diabetic platelet membrane and thus to modulate its hyperactivity.     

Activity of neutrophil NADPH oxidase in iron-deficient anemia

This study was designed to measure the effects of iron supplementation on respiratory burst in iron-deficient anemia. The performance of neutrophils was evaluated by measuring the activity of NADPH oxidase in 18 patients with iron-deficient anemia before and after body iron stores are saturated. The activity of NADPH oxidase was significantly lower in pretreatment patients relative to controls (p<0.05). The activity increased after iron supplementation to levels that had no significant differences relative to controls.     

Effects of Parenteral Supply of Iron on RBC Parameters,Performance, and Health in Neonatal Dairy Calves

The present study was designed to evaluate the effects of parenteral iron on red blood cell parameters, performance, and health in dairy Holstein calves. Twenty neonatal calves were equally divided at random into two groups, one of which served as controls. Care was taken to ensure homogeneity of sex, age, and general health status of the animals. The controls received a normal diet and water ad libitum, while the study animals were injected with 1 g iron as Fe-dextran 2 days after birth. A daily record was kept of the calves’ weight and growth parameters. At periods of 24–48 h after birth and at 14, 28, and 42 days of age, jugular blood was drawn from all the experimental and control animals to measure the packed cell volume, red blood count, hemoglobin, mean corpuscular hemoglobin, mean corpuscular hemoglobin concentration, and iron levels. At the start of the experiments, there were no significant differences between these parameters between the two study groups (p > 0.05). With time, significant differences were seen between most of the values measured (p < 0.05) except for the mean corpuscular hemoglobin, mean corpuscular hemoglobin concentration, and iron level. Significant differences were seen for total weight gain and mean daily weight gain, which were higher in the iron-supplemented group (p < 0.05).     

Effect of zinc deficiency and supplementation on lipid peroxidation of renal tissue in ovariectomized rats

The aim of this study was to investigate how zinc deficiency and supplementation affects lipid peroxidation in the renal tissue in ovariectomized rats. Four study groups were formed with 10 Spraque-Dawley rats each. Two of the groups served as normal and ovariectomized controls; the other two were ovariectomized rats that were zinc deficient and zinc supplemented, respectively. The zinc-deficient ovariectomized rats showed greater renal and plasma lipid peroxidation, as indicated by higher malondialdehyde levels than all other groups (p<0.05). These values were higher in the ovariectomized controls than those of the normal controls and of the ovariectomized, zinc-supplemented groups (p<0.05), which, in, turn, showed no significant differences of their respective renal and plasma malondialdehyde values. The renal and erythrocyte glutathione levels in the zinc-supplemented rats were higher than those in all other groups (p<0.05). The zinc-deficient group had the lowest renal and erythrocyte glutathione levels (p<0.05). The renal tissue zinc levels in the ovariectomized rats were higher than those in the zinc-deficient animals, but lower than in the normal controls and zincsupplemented rats (p<0.05). The zinc-supplemented animals had the highest renal tissue zinc levels (p<0.05). The results of this study suggest that zinc deficiency increases renal tissue damage in ovariectomized rats and that zinc supplementation can be used to prevent this condition.     

The effect of time of day and exercise on platelet functions and platelet–neutrophil aggregates in healthy male subjects

Platelet activation state changes by exercise. The effect of exercise time on platelet activation state and formation of platelet–neutrophil aggregates are not known yet. In this study the effect of exercise and time of day were examined on platelet activity with platelet–neutrophil aggregates. Ten moderately active males aged 27± 1.63 (mean±S.D.) years completed sub-maximal (70% VO_2max) exercise trials for 30 min. Blood pressure (BP) was recorded. Venous blood samples were obtained at rest, immediately post-exercise and after 30 min of recovery. Whole blood was analysed for haematocrit (Hct), haemoglobin (Hb), platelet count (PC), mean platelet count (MPV) and platelet aggregation (PA). Platelet–neutrophil aggregates and beta-thromboglobulin (β-TG) levels were assayed. Platelet count showed significant increase after morning exercise ((236± 32)×10⁹ l⁻¹ versus (202± 34)×10⁹ l⁻¹ baseline, p < 0.05). Exercise resulted in significantly lower MPV after the evening exercise (9.16± 0.5 fl versus 9.65± 0.36 fl, p < 0.05). Platelet aggregation by adenosine diphosphate (ADP) decreased after morning exercise and the recovery aggregation levels were significantly different at two different times of the day (68± 20% a.m. versus 80± 12% p.m., p < 0.05). It was also showed that platelet–neutrophil aggregates increased significantly from baseline after both exercises. Exercise-induced platelet–neutrophil aggregates were higher in the evening (10.7± 1.3% p.m. versus 6.4± 1.8% a.m., p < 0.0001). It is therefore concluded that besides platelet–platelet aggregation, exercise can cause platelet– neutrophil aggregates. In addition, time of day has an effect on platelet activation related events. Circadian variations of physiological parameters may have an effect on thrombus formation by platelet activation. (Mol Cell Biochem xxx: 119–124, 2005)     

Changes of total content of magnesium and zinc status in patients with chronic toxoplasmosis

The aim of this study was to investigate the effects of the Toxoplasma gondii infection on the total content of the essential elements Zn and Mg levels. Plasma zinc and magnesium concentrations were measured in 158 patients whose anti-T gondii IgG antibodies were positive. Scores were obtained for the patients and their age- and sex-matched 82 sero-negative healthy controls. The mean concentration of Mg in blood was significantly lower in sero-positive patients than in their controls both in females (p<0.01) and males (p<0.01). The average Zn concentration in seropositive female patients was 0.72±0.2 mg/L and 0.65±0.1 mg/L in controls (p>0.05). The mean values of the Zn in blood were 1.00±0.3 mg/L in seropositive male patients and 0.84±0.2 in controls (p>0.05). No correlation could be demonstrated between age and mean values of Zn and Mg in both sero-positive females/males and controls (p>0.05). No significant correlation could be found between blood Zn and Mg levels in seropositive female/male patients and controls (p>0.05). Mg levels were found to be clearly decreased in the patients with chronic toxoplasmosis, whereas no change was observed in Zn levels.     

Investigation of Serum Trace Element,Malondialdehyde and Immune Status in Drug Abuser Patients Undergoing Detoxification

Drug abuser patients (n = 104), age ranging from 19 to 42 years, were randomly recruited to investigate the serum levels of trace elements (Cu, Zn, Fe, and Mg), malondialdehyde (MDA), and immunoglobulin (IgG, IgA, and IgM) before and after clinical intervention. Control group also included 104 healthy individuals. Blood samples were analyzed for determining trace elements, MDA, and immunoglobulin using atomic absorption spectroscopy, Ultraviolet-Visible (UV-VIS) spectroscopy, and turbidimetry method, respectively. For serum level of Zn and Fe, the differences between the groups (before intervention, after intervention, and control) were not significant (p > 0.05). However, significant differences were found in serum copper levels between control group, drug abuser patients, and before and after intervention (p < 0.05). The concentration of Mg was found to be significantly higher (p = 0.007) in drug abuser patients than the controls, and after intervention, the level was restored to control value. A displacement of elemental homeostasis was observed in drug abuser patients compared to control, and it was improved after intervention. An increase in serum concentration of MDA was found in drug abuser patients compared to control subjects (p > 0.05) but was not statistically significant. After intervention, the concentration was restored to control value (p > 0.05). The serum concentrations of IgA and IgM were found to be significantly higher (p < 0.05) in drug abuser patients before intervention than the controls, and the level tended to be restored to control level after clinical intervention. Serum IgG level was found to be lower in drug abuser patients compared to controls and further declined significantly (p < 0.05) after intervention. These findings may suggest a possible imbalance in the levels of micronutrients, antioxidants, and immunoglobulin in drug abuser patients, which tend to be restored to control values after detoxification.     

Concentrations of seven trace elements in different hematological matrices in patients with type 2 diabetes as compared to healthy controls

This study aimed to compare the trace element status of patients with type 2 diabetes (n=53) with those of nondiabetic healthy controls (n=50). The concentrations of seven trace elements were determined in the whole blood, blood plasma, erythrocytes, and lymphocytes of the study subjects. Vanadium and iron levels in lymphocytes were significantly higher in diabetic patients as compared to controls (p<0.05 for iron and p<0.01 for vanadium). In contrast, lower manganese (p<0.01) and selenium (p<0.01) concentrations were detected in lymphocytes derived from patients with type 2 diabetes versus healthy subjects. Furthermore, significantly lower chromium levels (p<0.05) were found in the plasma of diabetic individuals as compared to controls. Trace element concentrations were not dependent on the degree of glucose control as determined by correlation analysis between HBA_1c versus metal levels in the four blood fractions. In summary, this study primarily demonstrated that trace element levels in lymphocytes of patients with type 2 diabetes could deviate significantly from controls, whereas, in general, no considerable differences could be found when comparing the other fractions between both patient groups. Therefore, it seems reasonable to analyze metal levels in leukocytes to determine trace element status in patients with type 2 diabetes and perhaps in other diseases.     

Plasma selenium levels in rheumatoid arthritis

The plasma selenium (Se) levels were determined in patients with rheumatoid arthritis (RA) and healthy controls. Plasma Se levels in 60 patients were found to be significantly lower than those in 60 normal, healthy controls (p<0.001). Similar significant differences were determined in sex-matched comparisons between patients and controls (p<0.001) but there was no significant difference in plasma Se levels in sex-matched comparisons in both groups (p>0.05). Our results suggest that Se is an important factor in RA.     

Mean platelet volume: Interrelation with platelet aggregation activity and glycoprotein IIb-IIIa and Ib expression levels

Increased mean platelet volume (MPV) is an independent risk factor of thrombotic events in patients with cardiovascular diseases. Interactions of MPV with platelet aggregation activity and contents of glycoprotein (GP) IIb-IIIa (αIIb/β3 integrin, fibrinogen receptor) and GP Ib (von Willebrand factor receptor) have been investigated in this study. The study was performed in a group of healthy volunteers (n = 38) and a group of patients with acute coronary syndrome (ACS, n = 116). Patient’s blood was collected at days 1, 3–5 and 8–12 after ACS development. All patients received acetylsalicylic acid (ASA, inhibitor of thromboxane A2 synthesis) as the antiaggregant therapy and most of them also received clopidogrel (ADP receptor antagonist), except 44 patients who had not taken clopidogrel at day 1 before first blood collection. Aggregation of volunteers’ platelets was stimulated by 1.25, 2.5, 5 and 20 μM ADP, while aggregation of patients’ platelets was stimulated by 5 and 20 μM ADP. GP IIb-IIIa and GP Ib content on the platelet surface was measured using ¹²⁵I-labelled monoclonal antibodies. GP IIb-IIIa and GP Ib genetic polymorphisms were determined in ACS patients. In healthy donors significant correlations between MPV and aggregation levels have been recognized at 1.25 μM and 2.5 μM ADP (correlation coefficient (r) values of 0.396 and 0.373, p < 0.05), while at 5 μM and 20 μM ADP these interactions did not reach the level of statistical significance (r values of 0.279 and 0.205, p > 0.05). Correlations between MPV and aggregation levels were observed at day 1 of ACS in a subgroup of patients receiving ASA but before the beginning of clopidogrel treatment (r values of 0.526, p < 0.001 and 0.368, p < 0.05 for 5 and 20 μM ADP, respectively). Correlations between these parameters were not found during combined treatment of patients with ASA and clopidogrel. Strong direct correlations between MPV and GP IIb-IIIa and GP Ib contents were detected in both healthy donors and ACS patients (at all time points): the r values ranged from 0.439 to 0.647 (p ≤ 0.001 for all correlations). Genetic polymorphisms of GP IIb-IIIa (GP IIIa Leu33Pro) and GP Ib ((?5)T/C (Kozak) and Thr145Met) identified in ACS patients did not affect expression levels of corresponding glycoproteins. The data obtained indicate that increased MPV values correlate with increased platelet aggregation activity and enhanced GP IIb-IIIa and GP Ib expression.     

Superoxide dismutase activity and zinc and copper concentrations in growth retardation

The aim of this study was to determine the effects of oral zinc treatment on red cell copper/zinc-superoxide dismutase (Cu/Zn-SOD) activity and zinc and copper concentrations in growth retardation. Thirty-five patients, average age of 11 yr, were selected. The control group consisted of 10 healthy children whose average age was 10 yr. Superoxide dismutase activity was determined by spectrophotometer. Copper and zinc concentrations were measured by an atomic absorption spectrophotometer. The activity of Cu/Zn-SOD before zinc treatment was higher than the controls (p<0.001). There was a decrease in Cu/Zn-SOD activity after zinc treatment (p<0.001) and the values after treatment were still higher than the controls (p<0.001). Plasma zinc concentrations before zinc treatment were lower than controls (p<0.01). After treatment, there was an increase in plasma zinc concentrations compared to controls and the patients' values before zinc treatment, respectively (p<0.001, p<0.001). After zinc treatment, plasma copper concentrations were decreased significantly (p<0.01). An increase in red cell zinc concentration (p<0.01) and a decrease in copper concentration (p<0.001), which were statistically significant, were seen after zinc treatment. The results suggested that there were significant alterations in Cu/Zn-SOD activity and zinc and copper concentrations during growth retardation. With zinc treatment, these parameters appeared to approach normal values.     

Synovial fluid and plasma selenium, copper, zinc, and iron concentrations in patients with rheumatoid arthritis and osteoarthritis

In recent years, a great number of studies have investigated the possible role of trace elements in the etiology and pathogenesis of rheumatoid arthritis (RA) and osteoartritis (OA). We studied synovial fluid and plasma concentrations of selenium (Se), zinc (Zn), copper (Cu), and iron (Fe) in patients with RA and OA and compared them with sex- and age-matched healthy subjects. Plasma albumin levels were measured as an index of nutritional status. Plasma Se, Cu, and Zn concentrations were determined by atomic absorption spectrophotometry and Fe concentrations were determined by the colorimetric method. Although plasma and synovial fluid Se concentration were found to be significantly lower (p<0.05, and p<0.05, respectively), Cu concentrations were significantly higher in patients with RA than those of healthy subjects and OA (p<0.05 and p<0.05, respectively). There were no significant differences in plasma and synovial fluid Zn concentrations and albumin levels among three groups (p>0.05). On the other hand, synovial fluid Cu and Fe concentrations were significantly higher in patients with OA than those of healthy subjects (p<0.05). There was a significantly positive correlation between synovial fluid Se−Cu values and Zn−Fe values in patients with RA. Our results showed that synovial fluid and plasma trace element concentrations, excluding Zn, change in inflammatory RA, but not in OA. These alterations in trace element concentrations in inflammatory Ra might be a result on the changes of the immunoregulatory cytokines.     

Serum zinc and magnesium levels in patients with blastocystosis

The aim of the study was to investigate the total content of the essential elements of zinc and magnesium levels in patients infected with Blastocystis hominis. Zinc and magnesium concentrations were measured in 52 patients who were positive for the intestinal parasite Blastocystis hominis. Scores were obtained for the positives and their age- and sex-matched 60 Blastocystis hominis-negative healthy controls. For comparison of two groups of continuous variables, the independent samples t-test was used. The mean concentration of magnesium in blood was significantly lower in Blastocystis hominis-positive patients than in their controls both in females (p<0.05) and males (p<0.05). The average zinc concentration in Blastocystis hominis-positive female patients was 0.61±0.2 mg/L and 0.60±0.2 mg/L in controls (p>0.05). The mean values of the zinc in blood were 0.62±0.2 mg/L in Blastocystis hominis-positive male patients and 0.82±0.1 in controls (p>0.05). No correlation could be demonstrated between age and mean values of zinc and magnesium in Blastocystis-positive females/males and controls (p>0.05). No significant correlation could be found between blood zinc and magnesium levels in Blastocystis-positive female/male patients and controls (p>0.05). Magnesium levels were found to be clearly decreased, whereas no change was observed in zinc levels in the patients with Blastocystis compared to controls.     

Trace element concentrations and superoxide dismutase and catalase activities in benign and malignant larynx tumors

The plasma and erythrocyte levels of zinc, copper, and magnesium and the activities of red-cell copper-zinc superoxide dismutase (Cu/Zn-SOD) and catalase (CAT) were determined in patients with benign and malignant tumors of the larynx. Blood samples from patients and healthy controls were drawn using heparinized tubes. The erythrocyte Cu/Zn-SOD and CAT activities were determined spectrophotometrically and the zinc, copper, and magnesium concentrations were determined in erythrocyte and plasma by atomic absorption spectrometry. Variance analysis was employed in the statistical evaluation of the findings. There was a significant increase in red-cell Cu/Zn-SOD activity in the subjects with malignant and benign tumors compared to controls (p<0.001). The CAT activity increased only in the benign tumor group (p<0.01). The plasma zinc concentrations were significantly lower in the malignant tumor group (p<0.05) and significantly higher in the benign tumor group (p<0.01). The erythrocyte copper concentrations were significantly lower in both benign and malignant tumor groups (p<0.001). The plasma copper and magnesium and the erythrocyte magnesium concentrations did not show significant differences relative to controls (p>0.05). The increases in the activities of SOD and CAT activities and the changes in trace elements concentrations can indicate the presence of increased reactive oxygen species that might play a part in the pathogenesis larynx tumors. Presented at the IX Asian-Pacific Congress of Clinical Biochemistry, March 9–14, 2002, New Delhi, India.     

Leptin Does Not Play a Major Role in Platelet Aggregation in Obesity and Leptin Deficiency

Objective: A recent study suggested that high concentrations of leptin enhance platelet aggregations. Therefore, the aim of this study was to investigate whether platelet aggregation is altered in patients with leptin gene mutations compared with obese subjects or controls. Research Methods and Procedures: Four men (one homozygous man and his three heterozygous brothers) carrying a leptin gene mutation; 20 age‐matched, healthy, unrelated men; and 18 age‐matched obese men were enrolled in the study. Adenosine diphosphate (ADP)‐, collagen‐, and epinephrine‐induced platelet aggregation were evaluated in all individuals. Results: Our results show that patients with the leptin gene mutation (both the homozygous and heterozygous patients) had significantly higher ADP‐induced (78.3 ± 3.4% vs. 57.9 ± 9.3%, p = 0.001), collagen‐induced (78.1 ± 2.9% vs. 56.7 ± 9.3%, p = 0.007), and epinephrine‐induced (76.5 ± 9.2% vs. 59.5 ± 7.70%, p = 0.003) platelet aggregation compared with controls. However, ADP‐, collagen‐, or epinephrine‐induced platelet aggregations were similar to those in obese patients. Platelet aggregation responses to a combination of pretreatment with leptin at concentrations of 20, 50, 100, or 500 ng/mL for 5 minutes and ADP at concentrations of 2 μmol/liter also were evaluated. However, we did not find significant increases in platelet aggregation even at high concentrations of leptin (100 or 500 ng/mL) in leptin‐deficient patients, obese subjects, or controls. Discussion: Our data show that similar to findings in obese humans, homozygous or heterozygous leptin deficiency is associated with increased platelet aggregation compared with controls, and that higher concentrations of leptin do not increase platelet aggregation.     

Responsivenes of total content changes of magnesium and zinc status in patients infected with Giardia intestinalis

The aim of the study was to investigate the changes of total content of the essential elements of zinc and magnesium levels in patients infected with Giardia intestinalis. Zinc and magnesium concentrations were measured in 64 patients who were positive for the intestinal parasite G. intestinalis. Scores were obtained for the positives and their 60 age- and sex-matched G. intestinalis-negative healthy controls. The mean concentration of magnesium in blood was significantly lower in G. intestinalis-positive patients than in their controls both in females (p<0.05) and males (p<0.05). The average zinc concentration in G. intestinalis-positive female patients was 0.76±0.3 mg/L and it was 0.60±0.2 mg/L in controls (p>0.05). The mean values of the zinc in blood were 0.73±0.2 mg/L in G. intestinalis-positive male patients and 0.82±0.1 mg/L in controls (p>0.05). No correlation could be demonstrated between age and mean values of zinc and magnesium in both G. intestinalis-positive females/males and controls (p>0.05). No significant correlation could be found between blood zinc and magnesium levels in G. intestinalis-positive female/male patients and controls (p>0.05). Magnesium levels were found to be clearly decreased, whereas no change was observed in zinc level in the patients infected with G. intestinalis compared to controls.     

Short‐Term Effects of Gastric Bypass Surgery on Circulating Ghrelin Levels

Objective: To prospectively evaluate the short‐term effects of Roux‐en‐Y gastric bypass (RYGBP) on ghrelin secretion and its relevance on food intake and body weight changes. Research Methods and Procedures: Ghrelin response to a standardized test meal was evaluated in eight obese patients (BMI, 43.5 to 59.1 kg/m²) before and 6 weeks after RYGBP. Ghrelin response was compared with that of an age‐matched group of six normal weight individuals (BMI, 19.6 to 24.9 kg/m²). Results: Fasting serum ghrelin levels were lower in obese subjects compared with controls (p < 0.05). Meal ingestion significantly suppressed ghrelin concentration in controls (p < 0.05) and obese subjects (p < 0.05), albeit to a lesser degree in the latter group (p < 0.05). Despite a 10.3 ± 1.5% weight loss, fasting serum ghrelin levels were paradoxically further decreased in obese subjects 6 weeks after RYGBP (p < 0.05). Moreover, at this time‐point, food intake did not elicit a significant ghrelin suppression. The changes in ghrelin secretion after RYGBP correlated with changes in insulin sensitivity (p < 0.05) and caloric intake (p < 0.05). Discussion: This study showed that the adaptive response of ghrelin to body weight loss was already impaired 6 weeks after RYGBP. Our study provides circumstantial evidence for the potential role of ghrelin in the negative energy balance in RYGBP‐operated patients.     

Longitudinal study of serum zinc and copper levels in hemodialysis patients and their relation to biochemical markers

A 6-mo longitudinal study of 48 hemodialysis patients (HPs) with chronic renal failure was performed. Three blood samplings were done. Samples of whole blood from each patient were collected during hemodialysis sessions after passing through the artificial kidney. Zinc and copper levels were measured by atomic absorption spectrometry. Additionally, 36 biochemical indexes were evaluated during the study. Fifty-two healthy matched controls were also considered. Mean serum zinc and copper concentrations in HPs were significantly decreased (Zn) and increased (Cu), when compared with healthy controls (p<0.01). Zinc concentrations found in the first and second blood samplings from patients were significantly lower than those measured for the third sampling (p<0.01). The etiology of chronic renal failure influenced the statistically serum Zn levels of patients (p<0.05). Serum copper levels of HPs were significantly diminished by the existence of secondary associated diseases (p<0.01). Uric acid and parathyroid hormone, and total-cholesterol and glutamic-pyruvic-transaminase levels were significantly (p<0.05) and linearly related with serum zinc and copper concentrations, respectively. From all of indexes, creatinine, direct bilirubin, magnesium, calcium, parathyroid hormone, transferrin, and albumin were statistically modified along the longitudinal study (p<0.05). Transferrin serum levels were significantly diminished in the third blood sampling, indicating the tendency toward anemia in the patients. This result is reinforced by low levels of biochemical and hematological indexes related with iron body staus.     

Iron-induced oxidative DNA damage in rat sperm cells in vivo and in vitro

We investigated whether acute iron intoxication causes oxidative DNA damage, measured in terms of 7-hydro-8-oxo-2′-deoxyguanosine, 8-oxodG, in nuclear DNA in testes and epididymal sperm cells in vivo and in vitro in rats. In addition, we investigated levels of the modified nucleoside in liver and kidney and measured its urinary excretion.

Sperm cells were isolated from the epididymides and the testes cells were isolated after homogenisation. In vitro, the sperm and testes cells were incubated with increasing concentrations of FeCl₂ ranging from 0 to 600 μM. The median (range) levels of 8-oxodG/10⁵ dG in the epididymal sperm cells increased from 0.48 (0.42–0.90) to 15.1 (11.4–17.6) (p < 0.05), whereas the level rose from 0.63 (0.22–0.81) to 8.8 (4.5–11.6) (p < 0.05) at 0 and 600 μM, respectively, in the testicular cells.

In vivo groups of 7–8 rats received 0, 200 or 400 mg iron/kg as dextran i.p. After 24h, epididymal sperm cells, testes, kidneys and liver were collected for analysis. Kidney and sperm DNA showed a significant increase in 8-oxodG in the iron-treated animals. The median (range) values of the 8-oxodG/10⁵ dG in the epididymal sperm cells rose from 0.66 (0.38–1.09) to 1.12 (0.84–5.88) (p < 0.05) at 0 and 400 mg iron/kg, respectively, whereas the values in the testes and liver showed no significant change. In the kidneys the 8-oxodG/10⁵ dG median (range) values were 0.98 (0.73–1.24), 1.21 (1.13–1.69) and 1.34 (1.12–1.66) after 0, 200 and 400 mg iron/kg, respectively (p < 0.05).

The 8-oxodG-excretion rate was measured in 24 h urine before and after iron treatment. The rate of urinary 8-oxodG excretion increased from 129 (104–179) pmol/24 h before treatment to 147 (110–239) pmol/24h after treatment in the group receiving 400 mg iron/kg (p < 0.05).

The results indicate that acute iron intoxication may increase oxidative damage to sperm and kidney DNA.