Interaction Between Iron Research Articles

Azomethine Fe3+ coordination compounds containing carbazole units: Synthetic approach, spectral characterization, and magnetic studies

The main aim of our study is to investigate various types of hybrid systems with a paramagnetic iron (III) ion as a core and photoactive derivatives of carbazole at the periphery. We have synthesized a variety of biligand coordination compounds with the composition [FeL2]A (where L is 2‐[2‐[(E)‐[4‐[4‐(3,6‐di‐tert‐butylcarbazol‐9‐yl)benzoyl]oxy‐2‐phenolate]methyleneamino]ethylamino]ethyl 4‐(3,6‐di‐tert‐butylcarbazol‐9‐yl)benzoate, A− is NO3− (I), Cl− (II), PF6− (III)). These systems were studied by superconducting quantum interference device (SQUID) magnetometry and X‐band electron paramagnetic resonance (EPR) spectroscopy. Magnetic measurements revealed mixed spin states (high spin [HS], low spin [LS]) of Fe (III) ions at room temperatures. Estimates of the corresponding spin contributions were made. It was found that all samples exhibit AFM exchange interactions between iron (III) ions. The ground spin state at 2.0 K was established and analyzed. EPR measurements confirmed the HS states of iron (III) and revealed two types of them: with weak distorted and strong low‐symmetry octahedral crystal fields.

Physicochemical Properties of Condensed Products of Interaction between Iron(II) Cations and Permanganate Ions

Condensed products are obtained via the partial oxidation of iron(II) cations using potassium permanganate, and their physicochemical properties are studied. Samples obtained at a potassium permanganate-to-iron(II) molar ratio close to stoichiometric in a preparation of magnetite have the greatest relative magnetic susceptibility. It is shown that the initial rate of formation of relative magnetic susceptibility in the range of 30–70°C depends linearly on the inverse thermodynamic temperature. Sedimentation of the synthesized products upon exposure to an external magnetic field is complete within 1–1.5 min. Diffractometry shows that crystalline magnetite samples form at a KMnO4 : Fe(II) molar ratio of 0.08–0.18. Magnetic granulometry reveals they have superparamagnetic properties. Under conditions of static sorption, the degree of extraction of chromate ions is 98.7%.

Insight into the interaction mechanism of iron ions with soil humic acids. The effect of the pH and chemical properties of humic acids.

The main aim of this work was to study the mechanisms of interaction between iron(II) ions and humic acids as a function of pH, iron concentration and various humic acids chemical properties, including the degree of humification, elemental composition, aromaticity and content of acidic functional groups. The results indicated that iron was bound by humic acids at pH 7 in amounts ∼2 times higher than at pH 5 (averaged capacities: 117 and 57 cmol/kg, respectively). Iron binding at pH 7 increased with increasing the total carboxylic and phenolic groups content and the degree of humification of humic acids (R-coefficients: 0.99 and 0.95, respectively). The stability of humic acid-iron complexes at pH 7 were only slightly lower than at pH 5 due to iron hydroxides formed at pH > 5 (averaged stability constants: 5.18 and 5.26, respectively). Iron coordination mode varied depending on pH: at pH 5, the bidentate (chelate) mode dominated, whereas at pH 7 the bridging mode appeared prevalent. The total amount of bound iron was much smaller than the content of the carboxylic and phenolic groups in humic acids, on average by ∼80 (pH 7) and ∼90.1% (pH 5) indicating the occurrence of steric effects in humic acid structure i.e. the reduction of the complexation capacity of free functional groups by adjacent groups occupied by iron and/or the formation of intramolecular aggregates with iron hindering the access of further metal ions. At pH 5 the complexes were soluble in the iron concentration range positively correlated to carboxylic and phenolic groups content, showing the protective nature of negatively charged functional groups on the stability of the solution. At this pH, the destabilization of the system was governed by the neutralization of humic acid charged structures by metal cations and the compression of the double electric layer. At pH 7 the stability of the humic acid-iron solution was largely determined by the form of iron, mainly by the precipitation of metal hydroxides acting as a flocculant destabilizing the solution by co-precipitation of humic acid-iron complexes.

State of Atoms and Interatomic Interactions in Complex Perovskite-Like Oxides: XXXVII.1 Synthesis and Magnetic Properties of SrTi1–xFe x O3 Solid Solutions

A series of SrTi1–xFe x O3 solid solutions was synthesized and characterized over a wide concentration range (x = 0.01–0.10). The magnetic susceptibility and magnetization data show that strong ferromagnetic interactions between iron(III) atoms exist in this system.

Theoretical analysis of the binding of iron(III) protoporphyrin IX to 4-methoxyacetophenone thiosemicarbazone via DFT-D3, MEP, QTAIM, NCI, ELF, and LOL studies.

Thiosemicarbazones display diverse pharmacological properties, including antimalarial activities. Their pharmacological activities have been studied in depth, but little of this research has focused on their antimalarial mode of action. To elucidate this antimalarial mechanism, we investigated the nature of the interactions between iron(III) protoporphyrin IX (Fe(III)PPIX) and the thione-thiol tautomers of 4-methoxyacetophenone thiosemicarbazone (MAPTSC). Dispersion-corrected density functional theory (DFT-D3), the quantum theory of atoms in molecules (QTAIM), the noncovalent interaction (NCI) index, the electron localization function (ELF), the localized orbital locator (LOL), and thermodynamic calculations were employed in this work. Fe(III)PPIX-MAPTSC binding is expected to inhibit hemozoin formation, thereby preventing Fe(III)PPIX detoxification in plasmodia. Preliminary studies geared toward the identification of atomic binding sites in the thione-thiol tautomers of MAPTSC were carried out using molecular electrostatic potential (MEP) maps and conceptual DFT-based local reactivity indices. The thionic sulfur and the 2 N-azomethine nitrogen/thiol sulfur of, respectively, the thione and thiol tautomers of MAPTSC were identified as the most favorable nucleophilic sites for electrophilic attack. The negative values of the computed Fe(III)PPIX-MAPTSC binding energies, enthalpies, and Gibbs free energies are indicative of the existence and stability of Fe(III)PPIX-MAPTSC complexes. MAPTSC-Fe(III) coordinate bonds and strong hydrogen bonds (N-H···O) between the NH2 group in MAPTSC and the C=O group in one propionate side chain of Fe(III)PPIX are crucial to Fe(III)PPIX-MAPTSC binding. QTAIM, NCI, ELF, and LOL analyses revealed a subtle interplay of weak noncovalent interactions dominated by dispersive-like van der Waals interactions between Fe(III)PPIX and MAPTSC that stabilize the Fe(III)PPIX-MAPTSC complexes.

Mössbauer study of the effect of pH on the rate of redox interactions between iron(III) and 4-n-hexylresorcinol in aqueous media

Mossbauer spectroscopy is used for a comparative study of the rate of iron(III) reduction by 4-n-hexylresorcinol (4-n-HR, a chemical analog of microbial autoregulators excreted by cells into the environment that allow intercellular communication) in aqueous media in the pH range of 1.5–4.5 simulating acidic soil conditions. The concomitant process of 4-n-HR oxidation is monitored using UV spectrophotometry.

Interactions between Iron(III)-hydroxide Polymaltose Complex and Commonly Used Drugs

Under physiological conditions, ferric ions are essentially insoluble because of the formation of polynuclear hydroxo-bridged complexes. Ferrous ions are more soluble but may produce hydroxyl radicals on reaction with hydrogen peroxide. Chelation of ferric and ferrous ions with organic ligands may prevent these undesirable reactions. Alternatively, iron(III)-hydroxide/oxide can be stabilized and solubilized by tight interactions with carbohydrates. The data presented in this work show that, because of its physicochemical properties, the iron(III)-hydroxide polymaltose complex (IPC, Maltofer) does not interact with the active ingredients of commonly used drugs such as acetylsalicylic acid (CAS 50-78-2), tetracycline hydrochloride (CAS 64-75-5), calcium hydrogen-phosphate (CAS 7757-93-9), methyl-L-dopa sesquihydrate (CAS 41372-08-1), and magnesium-L-aspartate hydrochloride (CAS 28184-71-6). In contrast, as confirmed by calculations using thermodynamic parameters, FeCl3 x 6H2O (CAS 10025-77-1) can form different types of complexes with these substances. Moreover, the data show that under aerobic conditions high concentrations of ascorbic acid (CAS 50-81-7) can lead to mobilization of iron from IPC and, thus, support the observation that orange juice slightly increases the uptake of iron from IPC.

Interactions between Iron(III)-hydroxide Polymaltose Complex and Commonly Used Medications

Simple iron salts, such as iron sulphate, often interact with food and other medications reducing bioavailability and tolerability. Iron(III)-hydroxide polymaltose complex (IPC, Maltofer) provides a soluble form of non-ionic iron, making it an ideal form of oral iron supplementation. The physicochemical properties of IPC predict a low potential for interactions. The effects of co-administration with aluminium hydroxide (CAS 21645-51-2), acetylsalicylic acid (CAS 50-78-2), bromazepam (CAS 1812-30-2), calcium acetate (CAS 62-54-4), calcium carbonate (CAS 471-34-1), auranofin (CAS 34031-32-8), magnesium-L-aspartate hydrochloride (CAS 28184-71-6), methyldopa sesquihydrate (CAS 41372-08-1), paracetamol (CAS 103-90-2), penicillamine (CAS 52-67-5), sulfasalazine (CAS 599-79-1), tetracycline hydrochloride (CAS 64-75-5), calcium phosphate (CAS 7757-93-9) in combination with vitamin D3 (CAS 67-97-0), and a multi-vitamin preparation were tested in rats fed an iron-deficient diet. Uptake of iron from radiolabelled IPC with and without concomitant medications was compared. None of the medicines tested had a significant effect on iron uptake. Iron-59 retrieval from blood and major storage organs was 64-76% for IPC alone compared with 59-85% following co-administration with other medications. It is concluded that, under normal clinical conditions, IPC does not interact with these medications.

π Back-Bonding of Iron(II) Complexes Supported by Tris(pyrid-2-ylmethyl)amine and Its Nitro-Substituted Derivatives

The electronic and geometric structures of a series of iron(II) complexes supported by tetradentate tris(pyrid-2-ylmethyl)amine-type ligands with different numbers of 4-nitropyridine groups, [(PyCH(2))(3-n)(4-NO(2)PyCH(2))(n)N] (n = 0-3), were examined by X-ray absorption fine-structure and variable-temperature (1)H NMR spectroscopies and theoretical calculations to reveal how the low-spin state is stabilized through π back-bonding interactions between iron(II) and 4-nitropyridine donor group(s).

Structure and magnetic properties of AgFeP 2O 7

AgFeP 2O 7 has been synthesized by flux crystallization and characterized by single crystal and powder X-ray diffraction (sp. gr. P2 1/ c, a=7.3298(2), b=7.9702(2), c=9.5653(2) Å, β=111.842(1)°, V=518.68(2) Å 3) and FTIR-spectroscopy. The structure is composed of isolated iron octahedra and phosphate tetrahedra interconnected into 3D network with hexagonal channels, where silver counter-ions are located. The magnetic behavior of the compound approaches the Curie–Weiss equation with a Weiss constant θ=−165.9 K indicating strong antiferromagnetic interaction between iron(III) ions.

Mechanisms of Bioleaching and the Visualization of these by Combined AFM & EFM

The understanding of mechanisms, by which microorganisms are effecting the dissolution of metal sulfides, what in general is called bioleaching, has progressed largely during the last 15 years. Whereas in the beginning of the nineties of the last century the direct and the indirect mechanisms were state of the knowledge, nowadays it is generally accepted that contact and non-contact mechanisms besides ferric iron, perhaps with some organic sulfhydryl groups, are the relevant players [1]. Consequently, since the introduction of these ideas/hypotheses, research in this field has been revived considerably, as is obvious from the literature[2]. Also molecular biology has started to contribute to the understanding of these processes due to the clarification of the basic reactions [3, 4, 5]. Summarizing the main elements of the contact and non-contact mechanisms, we know now that in case of the contact mechanism bacteria need to attach to metal sulfide surfaces by means of their extracellular, polymeric substances (EPS) due to a primary electrostatic interaction between iron(II)ions complexed by uronic acid residues. This first interaction is followed by hydrophobic interactions (more than half of the EPS of A. ferrooxidans are lipids) and perhaps covalent bonding (speculative). These forces are active in the Angstrom range (Å). Consequently, the space between bacterial outer membrane and the surface of a metal sulfide, which is filled by EPS, ranges between 10 and 50 nm only. In this space the dissolution reactions occur.

Aggregated Enhanced Raman Scattering in Fe(III)PPIX Solutions: The Effects of Concentration and Chloroquine on Excitonic Interactions

Resonance Raman spectra of hematin and hemin solutions are reported for 413 and 514 nm excitation wavelengths. Enhancement of A1g modes (1569 and 1370 cm(-1)) and B1g modes (1124 and 755 cm(-1)) as a function of increased concentration are observed when irradiating with 514 nm laser excitation but not 413 nm. This can be rationalized by considering an excitonic coupling mechanism. As the concentration of hematin increases there is an increased probability of supramolecular interactions between iron(III) protoporphyrin IX (Fe(III)PPIX) units occurring. The Fe(III)PPIX concentration reaches a saturation point in solution and excitonic coupling reaches a maximum causing the enhancement profile to plateau when applying 514 nm excitation. In contrast, when using 413 nm excitation there were no changes in band intensity with increased concentration showing that excitonic coupling through supramolecular interactions for aggregated solutions is wavelength dependent. Electronic absorption spectra show that as the concentration of Fe(III)PPIX increases in solution the Soret band is slightly blue shifted and the Q-band significantly broadens supporting the excitonic hypothesis. Understanding the mechanism that accounts for the Raman photophysical behavior of hemes at high concentrations provided an indirect method to monitor antimalarial drug interactions. A second aim was to investigate chloroquine binding to Fe(III)PPIX-OH/H2O monomers, pi-pi dimers and micro-oxo dimers formed in highly concentrated solutions approaching those of the digestive vacuole of the P. falciparum malaria parasite using excitonic Raman enhancement. It was hypothesized that the Raman excitonic enhancement mechanism could be impeded in heme aggregated solutions by the addition of chloroquine. This would result in a reduction in heme bands associated with the A1g modes including nu4. Resonance Raman spectra recorded using 514 nm excitation show that chloroquine (CQ) acts as a molecular spacer and binds noncovalently through dispersion interactions giving rise to pi-pi interactions, between micro-oxo dimer units of Fe(III)PPIX as evinced by the decrease in intensity of nu4 in the Raman spectrum as a function of increasing CQ mole ratio. In comparison, electronic spectra show that CQ can bind to the unligated face of Fe(III)PPIX-OH/H2O monomers, potentially reducing the formation of pi-pi dimers. This study has important implications in determining the effectiveness of potential antimalarial compounds that are thought to exert their effectiveness by binding through supramolecular interactions to the unligated faces of Fe(III)PPIX-OH/H2O monomers and micro-oxo dimers.

Synthesis of iron oxide nanoparticles of narrow size distribution on polysaccharide templates

We report here the preparation of nanoparticles of iron oxide in the presence of polysaccharide templates. Interaction between iron (II) sulfate and template has been carried out in aqueous phase, followed by the selective and controlled removal of the template to achieve narrow distribution of particle size. Particles of iron oxide obtained have been characterized for their stability in solvent media, size, size distribution and crystallinity and found that when the negative value of the zeta potential increases, particle size decreases. A narrow particle size distribution with D 100 = 275 nm was obtained with chitosan and starch templates. SEM measurements further confirm the particle size measurement. Diffuse reflectance UV-vis spectra values show that the template is completely removed from the final iron oxide particles and powder XRD measurements show that the peaks of the diffractogram are in agreement with the theoretical data of hematite. The salient observations of our study shows that there occurs a direct correlation between zeta potential, polydispersity index, bandgap energy and particle size. The crystallite size of the particles was found to be 30–35 nm. A large negative zeta potential was found to be advantageous for achieving lower particle sizes, owing to the particles remaining discrete without agglomeration.

Two novel dinuclear iron(III) complexes containing μ-oxo-di-μ-carboxylato motif

Two novel μ-oxo-di-μ-carboxylato-bridged iron(III) complexes of [Fe2(bpea)2(PhCO2)2(μ-O)] (ClO4)2·C2H5OH (1) and [Fe2(bpma)2(ClCH2COO)2(μ-O)](ClO4)2· H2O (2) (bpea = N,N-bis(2-pyridylmethyl)ethylamine, bpma = N,N-bis(2-pyridylmethyl)methylamine), have been synthesized and determined by X-ray diffraction. Complex (1) crystallizes in the Orthorhombic space group P nma with d(Fe···Fe) of 3.094 A and average d(Fe–Obbridge) of 1.805 A; Complex (2) crystallizes in the Monoclinic space group C 2/c, with d(Fe···Fe) of 3.109 A and average d(Fe–Obbridge) of 1.794 A. The magnetic studies indicate a stronger antiferromagnetic interaction between iron(III) ions through μ-oxo-di-μ-carboxylato-bridge for complex (1), with J = − 141.6 cm−1.

Studies of interaction between iron (III) and intermediates of synthetic neuromelanin by means of cyclic voltammetry of and dopamine

Neuromelanin is a complex polymer pigment found primarily in the dopaminergic neurons of the human substantia nigra, whose composition is complex including production of dopamine auto-oxidation, glutathione and a variety of amino acid. Neuromelanin forms stable complex with iron (III). We observed that 5,6-dihydroxyindole and its ramification possessed strong ability of chelating iron (III), and they are the production of dopamine auto-oxidation under physiological pH condition. In the present of l-cysteine, the relative yields of electrochemical oxidation of dopamine also had strong ability of chelating iron (III). The experimental results suggest that 5,6-dihydroxyindole and 5-S-cysteineldopamine play important roles in the process of synthetic neuromelanin chelating iron (III).

Tris(chloranilato)ferrate(III) Anionic Building Block Containing the (Dihydroxo)oxodiiron(III) Dimer Cation: Synthesis and Characterization of [(TPA)(OH)FeIIIOFeIII(OH)(TPA)][Fe(CA)3]0.5(BF4)0.5·1.5MeOH·H2O [TPA = tris(2-pyridylmethyl)amine; CA = chloranilate

[(TPA)(OH)FeIIIOFeIII(OH)(TPA)][Fe(CA)3]0.5(BF4)0.5.1.5MeOH.H2O (1) which possesses both the [FeIII(CA)3]3- (CA= chloranilate) and hydroxooxoiron(III) ions has had its structure determined by single-crystal X-ray diffraction. The 2-300 K magnetic susceptibility of 1 provides the magnetic parameters, g = 2.07, J/kB = -165 K (115 cm-1), theta = -1 K, and the spin impurity, rho = 0.05, which indicates a strong antiferromagnetic interaction between iron(III) ions via the oxo anion.

Electronic Structures of Six-Coordinate Ferric Porphyrin Complexes with Weak Axial Ligands: Usefulness of 13C NMR Chemical Shifts

1H NMR, (13)C NMR, and EPR spectra of six-coordinate ferric porphyrin complexes [Fe(Por)L2]ClO4 with different porphyrin structures are presented, where porphyrins (Por) are planar 5,10,15,20-tetraphenylporphyrin (TPP), ruffled 5,10,15,20-tetraisopropylporphyrin (TiPrP), and saddled 2,3,7,8,12,13,17,18-octaethyl-5,10,15,20-tetraphenylporphyrin (OETPP), and axial ligands (L) are weak oxygen ligands such as pyridine-N-oxide, substituted pyridine-N-oxide, DMSO, DMF, MeOH, THF, 2-MeTHF, and dioxane. These complexes exhibit the spin states ranging from an essentially pure high-spin (S = 5/2) to an essentially pure intermediate-spin (S = 3/2) state depending on the field strength of the axial ligands and the structure of the porphyrin rings. Reed and Guiset reported that the pyrrole-H chemical shift is a good probe to determine the spin state in the spin admixed S = 5/2,3/2 complexes (Reed, C. A.; Guiset, F. J. Am. Chem. Soc. 1996, 118, 3281-3282). In this paper, we report that the chemical shifts of the alpha- and beta-pyrrole carbons can also be good probes to determine the spin state because they have shown good correlation with those of the pyrrole-H or pyrrole-C(alpha). By putting the observed or assumed pyrrole-H or pyrrole-C(alpha) chemical shifts of the pure high-spin and pure intermediate-spin complexes into the correlation equations, we have estimated the carbon chemical shits of the corresponding complexes. The orbital interactions between iron(III) and porphyrin have been examined on the basis of these chemical shifts, from which we have found that both the d(xy)-a(2u) interaction in the ruffled Fe(T(i)PrP)L2+ and d(xy)-a(1u) interaction in the saddled Fe(OETPP)L2+ are quite weak in the high-spin and probably in the intermediate-spin complexes as well. Close inspection of the correlation lines has suggested that the electron configuration of an essentially pure intermediate-spin Fe(T(i)PrP)L2+ changes from (d(xy), d(yz))3(d(xy))1(d(z)2)1 to (d(xy))2(d(xz), d(yz))2(d(z)2)1 as the axial ligand (L) changes from DMF to MeOH, THF, 2-MeTHF, and then to dioxane. Although the DFT calculation has indicated that the highly saddled intermediate-spin Fe(OETPP)(THF)2+ should adopt (d(xy), d(yz))3(d(xy))1(d(z)2)1 rather than (d(xy))2(d(xz), d(yz))2(d(z)2)1 because of the strong d(xy)-a(1u) interaction (Cheng, R.-J.; Wang, Y.-K.; Chen, P.-Y.; Han, Y.-P.; Chang, C.-C. Chem. Commun. 2005, 1312-1314), our 13C NMR study again suggests that Fe(OETPP)(THF)2+ should be represented as (d(xy))2(d(xz), d(yz))2(d(z)2)1 because of the weak d(xy)-a(1u) interaction. The contribution of the S = 3/2 state in all types of the spin admixed S = 5/2,3/2 six-coordinate complexes has been determined on the basis of the (13)C NMR chemical shifts.

Ionically Crosslinked Alginate/Carboxymethyl Chitin Beads for Oral Delivery of Protein Drugs

Complex beads composed of alginate and carboxymethyl chitin (CMCT) were prepared by dropping aqueous alginate-CMCT into an iron(III) solution. The structure and morphology of the beads were characterized by IR spectroscopy and scanning electron microscopy (SEM). IR confirmed electrostatic interactions between iron(III) and the carboxyl groups of alginate as well as CMCT, and the binding model was suggested as a three-dimensional structure. SEM revealed that CMCT had a porous morphology while alginate and their complex beads had a core-layer structure. The swelling behavior, encapsulation efficiency, and release behavior of bovine serum albumin (BSA) from the beads at different pHs were investigated. The BSA encapsulation efficiency was fairly high (>90%). It was found that CMCT disintegrated at pH 1.2 and alginate eroded at pH 7.4 while the complex beads could effectively retain BSA in acid (>85%) and reduce the BSA release at pH 7.4. The results suggested that the iron(III)-alginate-CMCT bead could be a suitable polymeric carrier for site-specific protein drug delivery in the intestine.

Interactions between iron(III) and sucrose, dextran, or starch in complexes

This paper is aimed to study the nature of iron(III)–saccharide interactions in complexes and to look for new iron-containing drugs for supplementing organisms. Iron(III)-saccharide complexes were synthesized by reacting FeCl 3 and saccharides (rice starch, sucrose, and dextran) in basic solutions and their characteristics were compared with synthetic iron-oxide. Both crystalline and amorphous iron(III) complexes were obtained and identified by X-ray diffraction (XRD). Amorphous samples were chosen to investigate the interactions between iron(III) and saccharides by continuous-flow dissolution experiments. In acidic solution, the rate of iron release from amorphous iron complexes increased with the following order: iron-rice starch≈iron-oxide≪iron-sucrose≈iron-dextran. The colloidal structure of iron-rice starch was proposed to describe its similar dissolution profile with that of iron-oxide.

Reaction of Hydroxyurea with Iron(III): Products and the Stoichiometry of the Redox Reaction

AbstractProducts and reaction stoichiometry of the interaction between iron(III) and hydroxyurea (HU) in aqueous solutions were studied. Under condition of an excess of iron(III) over hydroxyurea, initially formed mono(hydroxyureato)iron(III) complex decomposed through redox processes, forming two moles of FeII, one mole of NH4+, one mole of CO2, and 0.5 mole of N2O per 1 mole of initially present HU. Hydroxyurea radical, H2N‐CO‐NHO?, was detected in the course of the reaction. The possible mechanism of the formation of the products is discussed and compared with that obtained for horseradish peroxidase mediated oxidation of hydroxyurea with hydrogen peroxide. The possibility of direct reaction of hydroxyurea with the iron center of protein R2 of the ribonucleotide reductase is proposed. The reaction examined may serve as a simple model for testing the suggested hydroxyurea like activity for chemical substances.