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PubMed1996

Expression and biochemical characterization of iron regulatory proteins 1 and 2 in Saccharomyces cerevisiae.

Iron-regulatory proteins (IRPs) 1 and 2 are cytosolic RNA-binding proteins that bind to specific stem-loop structures, termed iron-responsive elements (IREs) that are located in the untranslated regions of specific mRNAs encoding proteins involved in iron metabolism. The binding of IRPs to IREs regulates either translation or stabilization of mRNA. Although IRP1 and IRP2 are similar proteins in that they are ubiquitously expressed and are negatively regulated by iron, they are regulated by iron by different mechanisms. IRP1, the well-characterized IRP in cells, is a dual-function protein exhibiting either aconitase activity when cellular iron is abundant or RNA-binding activity when cellular iron is scarce. In contrast, IRP2 lacks detectable aconitase activity and functions exclusively as an RNA-binding protein. To study and compare the biochemical characteristics of IRP1 and IRP2, we expressed wild-type and mutant rat IRP1 and IRP2 in the yeast Saccharomyces cerevisiae. IRP1 and IRP2 expressed in yeast bind the IRE RNA with high affinity, resulting in the inhibition of translation of an IRE-reporter mRNA. Mutant IRP2s lacking a 73 amino acid domain unique to IRP2 and a mutant IRP1 containing an insertion of this domain bound RNA, but lacked detectable aconitase activity, suggesting that the presence of this domain prevents aconitase activity. Like IRP1, the RNA-binding activity of IRP2 was sensitive to inactivation by N-ethylmaleimide (NEM) or 5,5'-dithiobis(2-nitrobenzoic acid) (DTNB), indicating IRP2 contains a cysteine(s) that is (are) necessary for RNA binding. However, unlike IRP1, where reconstitution of the 4Fe-4S cluster resulted in a loss in RNA-binding activity, the RNA-binding activity of IRP2 was unaffected using the same iron treatment. These data suggested that IRP2 does not contain a 4Fe-4S cluster similar to the cluster in IRP1, indicating that they sense iron by different mechanisms.

Aconitate Hydratase↗
PubMed1977

[The metallic cations' capture through the external mucopolysaccharide surface coat encompassing the neurons (author's transl)].

Union of several iron compounds and other metallic cations to the duodenum mucines and to the neuronal surface mucopolysaccharides is studied. The results differentiate clearly the behaviour of the duodenum mucosubstances and neuronal surface mucosubstances. The duodenum mucines only attract to certain cations, in such a way that their union seems to be specific. On the contrary the neuronal surface mucopolysaccharides attract to most of the assaied metallic cations. These results coincide with other biochemical ones that prove that cationic attraction by the acid mucopolysaccharides is of electrostatic nature. To reveal the metallic cation union to the mucopolysaccharides is used ammonium sulphide in solution. The physiological role of these perineuronal mucopolysaccharides in relation to nervous impulse transmission and propagation is discussed attending to the obtained results.

Animals↗
PubMed2007

Combined effect of staining substances on the discoloration of esthetic Class V dental restorative materials.

The purpose of this study was to determine the combined effect of an organic substance (mucin as a substitute for salivary organic substances), chlorhexidine, and an iron compound/tea solution on the changes in the color of esthetic Class V dental restorative materials. Color of a glass ionomer, resin-modified glass ionomer, compomer and flowable resin composite of A2 shade, respectively, was determined according to the CIELAB color scale relative to the standard illuminant D65. Color was measured at baseline, and after sequential immersion in the following substances: Step-1, mucin in PBS (MCP) for 48 h; Step-2, chlorhexidine (CHX) for 24 h; Step-3, iron compound (IRN) or tea solution (TEA) up to 7 days; and Step-4, ultrasonic cleaning for 1 h. Color change (DeltaE(ab )*) was calculated by the equation: DeltaE(ab)* = [(DeltaL*)(2) + (Deltaa*)(2) + (Deltab*)(2)](1/2), of which DeltaL(*) indicates changes in value, Deltaa(*) indicates changes in red-green parameter and Deltab(*) indicates changes in yellow-blue parameter. DeltaE(ab)* values after immersion in MCP and CHX were compared, and DeltaE(ab)* values after immersion in IRN or TEA, and subsequent ultrasonic cleaning were compared with respect to the restorative material and immersion substance. DeltaE(ab)* and changes in the color parameters (DeltaL(*), DeltaC(ab)* and DeltaH(ab)*) were analyzed by repeated measures, analysis of variance and a post-hoc test at the 0.05 level of significance. Color changes after immersion in MCP were acceptable (DeltaE(ab)* < 3.3), and those after immersion in CHX were generally acceptable. The range of DeltaE(ab)* values after immersion in IRN was 3.1-19.6, and that after ultrasonic cleaning was 2.4-9.6. The range of DeltaE(ab)* values after immersion in TEA was 10.7-21.1, and that after ultrasonic cleaning was 11.9-14.5. Color changes of four Class V restorative materials after combined treatment with mucin, chlorhexidine and an iron compound/tea solution were not acceptable. Colors did not recover to their original values after ultrasonic cleaning. Modifications on the surface of a restoration should be considered to reduce stain accumulation.

Acrylic Resins↗
PubMed1975

Oxidation of dicyano-bis(1,10 phenanthroline) iron(II) by compounds I and II of cytochrome c peroxidase.

The apparent bimolecular rate constant for the oxidation of dicyano-bis(1,10 phenanthroline) iron(II) by compound II of cytochrome c peroxidase (ferrocytochrome c; hydrogen-peroxide oxidoreductase EC 1.11.1.5) has been measured over the pH range 2.5-11.0 at 0.1 M ionic strength, 25 degrees C, by the stopped-flow technique. An ionizable group in the enzyme, with a pKa of 4.5, strongly influences the electron transfer rate between the ferrous complex and the oxidized site in the enzyme. The electron transfer is fastest when the group is protonated, with a rate constant of 2.9 - 10-5 M--1 - s-1. The rate constantdecreases over three orders of magnitude when the proton dissociates. The apparent bimolecular rate constant for the oxidation of the ferrous complex by compound I of cytochrome c peroxidase was determined between pH 3.5 and 6. Under all conditions where this rate constant could be measured it was about three times larger than that for the oxidation by compound II.

Cytochrome c Group↗
PubMed1996

Magnetic resonance imaging reveals micro-haemorrhage in the feet of diabetic patients with a history of ulceration.

Soft tissue haemorrhage in the foot is a possible precursor of ulceration in patients with diabetic peripheral neuropathy. High resolution 'targetted' magnetic resonance imaging was used to scan the forefoot. Neuropathic patients with and without previous ulceration were matched for degree of neuropathy, mean vibration perception threshold 33.5 +/- 4.2 V (previous ulcer) vs 31.0 +/- 6.9 V (no ulcer), age, sex, and duration of diabetes against non-neuropathic controls. There were nine patients in each category. Paramagnetic materials, e.g. iron compounds, cause a signal void ('drop-out') on gradient-echo images which disappear on spin-echo images. Evidence of haemorrhage was seen in 6 patients with previous ulceration, and none in the other groups (p = 0.009, chi square test). Autologous injection of 20 microliters of blood into the foot of a healthy volunteer produced similar images, a 'drop-out' 1 cm across being visible on magnetic resonance scanning 3 days later. Peak vertical forefoot pressures were not significantly different in the neuropathic groups 0.67 +/- 0.20 vs 0.60 +/- 0.13 Pa but were lower in the non-neuropathic group, 0.43 +/- 0.11 Pa (p = 0.0004, Mann-Whitney), and do not explain the appearance of these haemorrhages. Magnetic resonance imaging provides a sensitive way of detecting micro-haemorrhage and its presence may predict an increased risk of foot ulceration.

Blood Pressure↗
PubMed1990

A protein-free medium for the growth of hybridomas and other cells of the immune system.

A protein-free medium, termed ABC, has been developed which essentially eliminates the need for serum proteins. ABC supports the long-term growth of murine hybridomas as well as other transformed cells of the immune system. The requirement of hybridoma growth for transferrin has been met by substituting the soluble organo-iron compound, sodium nitroprusside. Substantial improvement in the growth of hybridomas was afforded by the inclusion of 18 trace elements complexed to disodium ethylene diaminetetraacetate (EDTA). The medium was further improved by the inclusion of components not found in Ham's F12 medium or by raising the concentrations of existing low molecular weight components. Murine hybridomas can be cultured routinely in this protein-free medium in an anchorage-independent manner with doubling times generally under 24 h. Visualized on electrophoretic gels, levels of monoclonal antibody taken from those cultures often exceeded 80% of the total protein. The medium was also able to support the growth of HuT 78 and H9 cells as well as certain other transformed cells of the immune system. In addition, normal human peripheral blood lymphocytes, activated with phytohemagglutinin and cultured with 50 U/ml recombinant interleukin 2, could be grown for 2 wk with a 50-fold expansion over input cell number.

Amino Acids↗
PubMed2000

The magnetic resonance imaging of musculoskeletal hemorrhage.

Hematomas in the extremities can present clinically as a soft tissue mass. Hematomas can usually be distinguished from neoplasia on MR by the signal patterns of hemoglobin breakdown products, which are dependent on the chemical bonding and oxidation state of hemoglobin iron. Beginning with a discussion of relevant atomic electronic structure, this review will examine how oxyhemoglobin, deoxyhemoglobin, methemoglobin, and hemosiderin, the principal iron compounds occurring in the various stages of a hematoma, affect its appearance on MRI.

Diagnosis, Differential↗
PubMed2001

Adhesion of the dissimilatory Fe(III)-reducing bacterium Shewanella alga BrY to crystalline Fe(III) oxides.

Shewanella alga BrY adhesion to hydrous ferric oxide, goethite, and hematite was examined. Adhesion to each oxide followed the Langmuir adsorption model. No correlation between adhesion and Fe(III) oxide surface area or crystallinity was observed. Zeta potential measurements suggested that electrostatic interactions do not influence S. alga BrY adhesion to these minerals. Cell adhesion does not appear to explain the recalcitrance of crystalline Fe(III) oxides to bacterial reduction.

Bacterial Adhesion↗
PubMed1983

The interaction between heme and protein in cytochrome c1.

The optical spectrum of reduced bovine cytochrome c1 at 77 K shows a fine splitting of the beta-band, which is indicative of the native conformation of the protein. At room temperature, this conformation is reflected in an absorbance band at 530 nm. The exposure of the heme of ferrocytochrome c1, investigated by means of solvent-perturbation spectroscopy, appears to be extremely sensitive to temperature and SH reagents bound to the oxidized protein. Addition of combinations of potential ligands to the isolated tryptic heme peptide of cytochrome c1 reveals that only a mixture of methionine and cysteine (or their equivalents) generates a beta-band at 77 K which is identical in shape to that of native cytochrome c1. In the EPR spectrum of a complex of ferrocytochrome c1 and nitric oxide at pH 10.5, no hyperfine splitting derived from a second ligated nitrogen atom could be detected. The results indicate that methionine and cysteine are the axial ligands of heme in cytochrome c1. The EPR spectrum of isolated ferricytochrome c1 is that of a low-spin heme iron compound with a gz value of 3.36 and a gy value of 2.04.

Animals↗
PubMed1985

Modulation of interleukin 2 release from a primate lymphoid cell line in serum-free and serum-containing media.

The ability to grow a clone of the cell line, MLA144, which is a constitutive producer of interleukin 2 (IL-2), in serum-free medium permitted the study of the direct effect of various agents on cell growth and IL-2 production in a homogeneous population. Bovine serum albumin (BSA) at 4 mg/ml was optimal for cell growth and IL-2 production. Selenium at 10 ng/ml enhanced IL-2 production nearly twofold and lithium at 42 ng/ml also enhanced IL-2 production by nearly twofold. Neither compound at these levels altered cellular proliferation. Two other compounds, iron and zinc, known to be associated with cellular proliferation and/or immunoregulation did not alter IL-2 production. Catalase or horseradish peroxidase was able to substitute for BSA and maintain the long-term growth of the MLA144 clone with only a 30% decrease in the rate of cellular proliferation and a 50% decrease in IL-2 production compared to cells maintained in the serum-free formulation with BSA. Addition of 0.5 mg of BSA to the catalase serum-free formulation increased the production of IL-2 to 70% of that of cells cultured in the BSA-containing serum-free formulation. The catalase-containing serum-free formulation has the advantage of consisting of only three proteins, catalase, insulin, and transferrin, at a very low protein content. The catalase-containing serum-free medium also supported the long-term growth of a human T-cell line, HSB-2.

Animals↗
PubMed1996

Prozac: panacea or puzzle?

Selective 5-HT (serotonin) reuptake inhibitors (SSRIs) lack many of the adverse side-effects of older antidepressants. One of these compounds, fluoxetine (Prozac), has acquired manifest public awareness that is not evident with other SSRIs. Here, Clare Stanford compares the basic pharmacology of SSRIs to determine whether fluoxetine has any atypical features, and thus provide scientific justification for the attention focused on this compound. Ironically, fluoxetine might be distinctive in that it is the least selective SSRI and has marked effects on catecholamine function in the brain.

Animals↗
PubMed2006

Design of molecular wires based on supramolecular structures for application in glucose biosensors.

In the present work, the synthesis and the spectroelectrochemical characterization of a novel iron compound derived of tetra-2-pyridyl-1,4-pyrazine (TPPZ) with hexacyanoferrate species forming a very stable supramolecular complex in the presence of polypyrrole (PPY) matrix, is described. The hybrid material has shown excellent catalytic activity towards H(2)O(2) detection that makes it suitable for being used as redox mediator in a glucose biosensor. The hybrid FeTPPZFeCN/PPY film presents satisfactory detection limits and high sensitivity for H(2)O(2) in the presence of K(+) or Na(+) ions. For the glucose biosensor, a linear range up to 1.1 mmoll(-1) of glucose was observed with no interferences. In this case, the sensitivities obtained were 7.88 and 5.90 microAmmol(-1)lcm(-2) in phosphate buffer or NaCl solutions, respectively. The good sensitivity is related to the presence of a high-dimensional structure based on polypyridine type ligands providing an "electron antennae effect" facilitating electron tunneling between the protein and the electrode.

Biosensing Techniques↗
PubMed2005

Retention of tannic acid and condensed tannin by Fe-oxide-coated quartz sand.

This paper intends to shed light on the interactions between tannin and mineral soil particles. For that purpose, aqueous solution of condensed tannin (CT) (derived from Black pine (Pinus nigra var. maritima)) and commercially available tannic acid (TA) were added to purified quartz (Qtz) sand and quartz sand coated with either goethite (Gt) or ferrihydrite (Fh). After solvent removal by evaporation the samples were extracted by water. The extracts were analysed for organic carbon, total phenolics and CT. The extractability of the two tannins was small and increased in the order Qtz-Fh < Qtz-Gt < Qtz. For all mineral samples, TA was more extractable than CT. Bonding of tannins to the mineral samples and the partial peptisation of the Fe oxide coatings upon the binding resulted in complex tannin release curves. Our results suggest that the inextractability of tannins from natural soils and the absence of tannins in soil leachates might be caused by strong adsorption on soil minerals such as Qtz and Fe (oxy)(hydr)oxides. The results of competition experiments with mixtures of both tannins demonstrate that the CTs, and TA in particular, can release large amounts of Fe (oxides), suggesting that the tannins are excellent metal-mobilising agents. We therefore suggest that the fate of tannins in the mineral soil environment is highly dependent on the abundance of weakly bonded secondary oxides.

Adsorption↗
PubMed2006

Influence of kinetics on the determination of the surface reactivity of oxide suspensions by acid-base titration.

The effect of acid-base titration protocol and speed on pH measurement and surface charge calculation was studied on suspensions of gamma-alumina, hematite, goethite, and silica, whose size and porosity have been well characterized. The titration protocol has an important effect on surface charge calculation as well as on acid-base constants obtained by fitting of the titration curves. Variations of pH versus time after addition of acid or base to the suspension were interpreted as diffusion processes. Resulting apparent diffusion coefficients depend on the nature of the oxide and on its porosity.

Aluminum Oxide↗