Ultraviolet A radiation induces immediate release of iron in human primary skin fibroblasts: the role of ferritin.
(25/3222)
In mammalian cells, the level of the iron-storage protein ferritin (Ft) is tightly controlled by the iron-regulatory protein-1 (IRP-1) at the posttranscriptional level. This regulation prevents iron acting as a catalyst in reactions between reactive oxygen species and biomolecules. The ultraviolet A (UVA) radiation component of sunlight (320-400 nm) has been shown to be a source of oxidative stress to skin via generation of reactive oxygen species. We report here that the exposure of human primary skin fibroblasts, FEK4, to UVA radiation causes an immediate release of "free" iron in the cells via proteolysis of Ft. Within minutes of exposure to a range of doses of UVA at natural exposure levels, the binding activity of IRP-1, as well as Ft levels, decreases in a dose-dependent manner. This decrease coincides with a significant leakage of the lysosomal components into the cytosol. Stabilization of Ft molecules occurs only when cells are pretreated with lysosomal protease inhibitors after UVA treatment. We propose that the oxidative damage to lysosomes that leads to Ft degradation and the consequent rapid release of potentially harmful "free" iron to the cytosol might be a major factor in UVA-induced damage to the skin. (+info)
Recurrent mutations in the iron regulatory element of L-ferritin in hereditary hyperferritinemia-cataract syndrome.
(26/3222)
BACKGROUND AND OBJECTIVE: Hereditary hyperferritinemia-cataract syndrome (HHCS) is an autosomal dominant disorder characterized by bilateral cataracts and increased serum and tissue L-ferritin, in the absence of iron overload. The deregulation of ferritin production is caused by heterogeneous mutations in the iron regulatory element (IRE) of L-ferritin that interfere with the binding of iron regulatory proteins. DESIGN AND METHODS: We have identified several patients from three unrelated Italian families with HHCS. Iron parameters were assessed by standard methods. The IRE element of L-ferritin was amplified by PCR using appropriate primers and directly sequenced. RESULTS: Ferritin levels ranged from 918 microg/L to 2490 microg/L in the patients studied. In one family bilateral cataracts were diagnosed early in life, whereas in the others cataracts were diagnosed around 40-50 years. The female proband of family 3 presented with a severe iron deficiency anemia, which was unrecognized because of the increased ferritin values. Sequencing of the IRE element of L-ferritin in the probands of the three families identified three different nucleotide substitutions (+32 GAE A, +40 AAE G and +39 CT) in the IRE of L-ferritin. These mutations have already been reported in unrelated subjects of different ethnic origins. INTERPRETATION AND CONCLUSIONS: Our findings are consistent with recurrent mutations associated with HHCS and underline the importance of this syndrome in the differential diagnosis of unexplained hyperferritinemia. In addition, the findings highlight the role played by transferrin saturation in the diagnosis of iron deficiency in these patients. (+info)
Bacterioferritin A modulates catalase A (KatA) activity and resistance to hydrogen peroxide in Pseudomonas aeruginosa.
(27/3222)
We have cloned a 3.6-kb genomic DNA fragment from Pseudomonas aeruginosa harboring the rpoA, rplQ, katA, and bfrA genes. These loci are predicted to encode, respectively, (i) the alpha subunit of RNA polymerase; (ii) the L17 ribosomal protein; (iii) the major catalase, KatA; and (iv) one of two iron storage proteins called bacterioferritin A (BfrA; cytochrome b1 or b557). Our goal was to determine the contributions of KatA and BfrA to the resistance of P. aeruginosa to hydrogen peroxide (H2O2). When provided on a multicopy plasmid, the P. aeruginosa katA gene complemented a catalase-deficient strain of Escherichia coli. The katA gene was found to contain two translational start codons encoding a heteromultimer of approximately 160 to 170 kDa and having an apparent Km for H2O2 of 44.7 mM. Isogenic katA and bfrA mutants were hypersusceptible to H2O2, while a katA bfrA double mutant demonstrated the greatest sensitivity. The katA and katA bfrA mutants possessed no detectable catalase activity. Interestingly, a bfrA mutant expressed only approximately 47% the KatA activity of wild-type organisms, despite possessing wild-type katA transcription and translation. Plasmids harboring bfrA genes encoding BfrA altered at critical amino acids essential for ferroxidase activity could not restore wild-type catalase activity in the bfrA mutant. RNase protection assays revealed that katA and bfrA are on different transcripts, the levels of which are increased by both iron and H2O2. Mass spectrometry analysis of whole cells revealed no significant difference in total cellular iron levels in the bfrA, katA, and katA bfrA mutants relative to wild-type bacteria. Our results suggest that P. aeruginosa BfrA may be required as one source of iron for the heme prosthetic group of KatA and thus for protection against H2O2. (+info)
Radioimmunoassay of human cardiac acidic isoferritin: a new index for hepatic cancer.
(28/3222)
OBJECTIVE: To investigate human cardiac acidic isoferritin as a specific index of hepatic cancer. METHODS: Acidic isoferritin was isolated and purified from human heart muscle. A radioimmunoassay for the acidic isoferritin in human serum has been developed, on the equilibrium method. The antiserum was obtained from rabbits immunized with purified acidic isoferritin. The 125I-acidic isoferritin was prepared by the chloramine-T method. The data were processed using the automated smoothed spline function data processing program. RESULTS: The intra- and inter-assay CV of acidic isoferritin RIA were 1.65% and 9.71%, respectively, and the recovery rate was 102%. The antiserum provided a linear response from 7.0 to 369.6 micrograms/L with ED50 of 27.50 micrograms/L. The cross reactivity with AFP, CEA, lactoferrin and transferrin was negligible, and that with ferritin was 1.74%. The serum acidic isoferritin concentration showed a considerable variation in different sex and age groups. The serum acidic isoferritin was measured in liver diseases including hepatic cancer, hepatic cirrhosis and acute and chronic hepatitis. Its sensitivity for diagnosis of hepatic cancer was 73.05%, independent from the severity of hepatic injury. In 8 malignant tumors studied, acidic isoferritin appeared the most valuable in the diagnosis of hepatic cancer, with its positive, negative, false positive and false negative rates all being ideal. CONCLUSIONS: Acidic isoferritin may turn to be a rather specific index of hepatic cancer. Combination of monitoring both acidic isoferritin and AFP would raise the positive detection and specificity in the diagnosis of hepatic cancer. (+info)
Iron nutritional status in preterm infants fed formulas fortified with iron.
(29/3222)
AIMS: To prospectively evaluate the iron nutritional status of preterm infants fed either a term (0.5 mg/dl iron) or preterm (0.9 mg/dl) formulas fortified with iron after hospital discharge. METHODS: Healthy low birthweight preterm infants were randomly assigned into three groups at the time of hospital discharge. Group A were fed an iron fortified preterm formula (0.9 mg/dl iron) until 6 months corrected age; group B, a fortified term formula (0.5 mg/l iron) until 6 months corrected age group C, the preterm formula between hospital discharge and term, then the term formula until 6 months corrected age. RESULTS: Seventy eight infants were followed up to 6 months corrected age. Iron intake from formula differed significantly between the groups (A, 1.17 mg/kg/day (SD 0.32) > C, 0. 86 mg/kg/day (SD 0.40) = B, 0.81 mg/kg/day (SD 0.23); p < 0.0001). Haemoglobin concentrations were similar to those of iron sufficient preterm infants of the same postnatal age, and term infants of the same postmenstrual age (after 3 months of age). There were no significant differences in haemoglobin concentration (p = 0.391), plasma ferritin (A vs B, p = 0.322), or in the incidence of iron deficiency (A vs B, p = 0.534). CONCLUSIONS: Iron fortified formulas containing between 0.5 and 0.9 mg/dl iron seem to meet the iron nutritional needs of preterm infants after hospital discharge. (+info)
The influence of epitope availability on atomic-force microscope studies of antigen-antibody interactions.
(30/3222)
The ability of the atomic-force microscope (AFM) to detect interaction forces between individual biological molecules has recently been demonstrated. In this study, force measurements have been obtained between AFM probes functionalized with the beta-subunit of human chorionic gonadotrophin (betahCG) and surfaces functionalized with anti-betahCG antibody. A comparison of the obtained results with previous anti-ferritin antibody-binding data identifies differences when the antigen molecule expresses only a single epitope (betahCG), rather than multiple epitopes (ferritin), for the monoclonal antibodies employed. Specifically, the probability of observing probe-sample adhesion is found to be higher when the antigen expresses multiple epitopes. However, the periodic force observed in the adhesive-force distribution, due to the rupture of single antigen-antibody interactions, is found to be larger and more clearly observed for the mono-epitopic system. Hence, these findings indicate the potential of the AFM to distinguish between multivalent and monovalent antibody-antigen interactions, and demonstrate the influence of the number of expressed epitopes upon such binding studies. (+info)
Protein adhesion force dynamics and single adhesion events.
(31/3222)
Using the manipulation force microscope, a novel atomic force microscope, the adhesion forces of bovine serum albumin, myoglobin, ferritin, and lysozyme proteins to glass and polystyrene substrates were characterized by following the force necessary to displace an adsorbed protein-covered microsphere over several orders of magnitude in time. This force was consistent with a power law with exponent a = 0.37 +/- 0.03 on polystyrene, indicating that there is no typical time scale for adhesion on this substrate. On glass, the rate of adhesion depended strongly on protein charge. Forces corresponding to single protein adhesion events were identified. The typical rupture force of a single lysozyme, ferritin, bovine serum albumin, and myoglobin protein adhering to glass was estimated to be 90 +/- 10 pN, 115 +/- 13 pN, 277 +/- 44 pN, and 277 +/- 44 pN, respectively, using a model of the experimental system. These forces, as well as the force amplitudes on hydrophobic polystyrene, correlate with protein stiffness. (+info)
Manganese absorption and retention by young women is associated with serum ferritin concentration.
(32/3222)
BACKGROUND: The interaction between iron and manganese in the gut is well characterized but iron status has not been shown to affect manganese absorption. OBJECTIVE: The objective of this study was to determine whether iron status as determined by serum ferritin concentrations affects manganese absorption, retention, balance, and status. DESIGN: The subjects were healthy young women; 11 had serum ferritin concentrations >50 microg/L and 15 had serum ferritin concentrations <15 microg/L. In a crossover design, subjects consumed diets that supplied either 0.7 or 9.5 mg Mn/d for 60 d. Manganese absorption and retention were assessed during the last 30 d of each dietary period by using an oral dose of 54Mn; balance was assessed simultaneously. RESULTS: Dietary manganese did not affect manganese status, but high serum ferritin depressed arginase activity. The interaction of ferritin status and dietary manganese affected 54Mn absorption and biological half-life. Absorption was greatest in subjects with low ferritin concentrations when they were consuming the low-manganese diet, and was least in subjects with high ferritin concentrations. Biological half-life was longest when subjects with high ferritin concentrations consumed the low-manganese diet, and was shortest in all subjects consuming the high-manganese diet. Manganese balance was only affected by the amount of manganese in the diet. CONCLUSIONS: These results show that iron status, as measured by serum ferritin concentration, is strongly associated with the amount of manganese absorbed from a meal by young women. When greater amounts of manganese are absorbed, the body may compensate by excreting manganese more quickly. (+info)