Polymerization Mode Research Articles

Modification of novolac resin by interpenetrating network formation with poly(butyl acrylate)

AbstractInterpenetrating networks (IPNs) of novolac (phenol formaldehyde) resin and poly(butyl acrylate) (PBA) were prepared by a sequential mode of polymerization. Both full IPNs and semi‐IPNs of different compositions were synthesized and characterized with respect to their mechanical properties, that is, their modulus, ultimate tensile strength (UTS), elongation‐at‐break percentage, and toughness. Their thermal properties were examined with differential scanning calorimetry and thermogravimetric analysis (TGA). A morphological study was performed with an optical microscope. The effects of the variation of the blend ratios on the aforementioned properties were studied. There was a gradual decrease in the modulus and UTS with a simultaneous increase in the elongation‐at‐break percentage and toughness for both types of IPNs as the proportions of PBA were increased. With increasing proportions of PBA, the glass‐transition temperatures of the different IPNs underwent shifts toward a lower temperature region. This showed a plasticizing influence of PBA on the rigid and brittle phenolic matrix. TGA thermograms depicted the classical two‐step degradation for the phenolic resin. Although there was an apparent increase in the thermal stability at the initial stage (up to 350°C), particularly at lower temperatures, a substantial decrease in the thermal stability was observed at higher temperatures under study. In all the micrographs of full IPNs and semi‐IPNs, two‐phase structures were observed, regardless of the PBA content. © 2005 Wiley Periodicals, Inc. J Appl Polym Sci 97: 2407–2417, 2005

Hardness and diametral tensile strength of a hybrid composite resin polymerized with different modes and immersed in ethanol or distilled water media

The aim of this in vitro study was to evaluate the microhardness and diametral tensile strength of a hybrid composite resin (Z250, 3M ESPE) polymerized with four different modes of light exposure and immersed in two different media. Composite resin specimens were randomly polymerized according to the experimental groups (conventional, 550 mW/cm(2)/30 s; soft start, 300 mW/cm(2)/10 s + 550 mW/cm(2)/20 s; high intensity, 1060 mW/cm(2)/10 s; pulse delay, 550 mW/cm(2)/1 s + 60 s of waiting time + 550 mW/cm(2)/20 s) and immersed in one of two media (distilled water or absolute ethanol) for 24 h. After that, microhardness (M) and diametral tensile strength (DTS) tests were performed. For DTS, there were no statistical differences among the polymerization modes, however, ethanol medium groups presented statistically lower DTS (p < 0.05) than water medium. For the M test, samples immersed in ethanol medium presented lower M for almost all groups. Conventional mode presented higher M values for the groups immersed in water medium. In ethanol medium, conventional and pulse delay groups presented higher M values, statistically different (p < 0.05) from the high intensity group. For all experimental conditions, the top surface showed higher M than the bottom surface. Different polymerization modes can be related to the different polymer structures formed, and consequently with different physical properties of resin composite. The immersion media can alter the physical properties of resin composites of different polymer structures.

Heterogenized “Ligand-Free” Lanthanide Catalysts for the Homo- and Copolymerization of Ethylene and 1,3-Butadiene

Silica-supported lanthanide catalysts have been prepared from the reaction of dehydroxylated silica and tris(bistrimethylsilyl)amido lanthanides Ln[N(SiMe3)2]3 (Ln = Sc, Y, La, Nd, Sm, Gd, Dy). On activation with AliBu3 (TIBA), the compounds catalyze the polymerization of ethylene, with the scandium catalyst showing the highest activity. There is an inverse relationship between ionic radius of the lanthanide metal and the activity of the resulting catalyst. The polymer produced was highly linear and of high molecular weight. Activation with magnesium alkyls was less effective but gave polyethylene of slightly higher molecular weight. The supported lanthanides, when activated with TIBA, were also active for the polymerization of 1,3-butadiene, with very high molecular weight polymers being produced. In all cases the polymer was found to be predominately 1,4-cis (typically 85−90% by diad analysis), with only scandium producing a much higher level of 1,4-trans units. The system was also shown to copolymerize ethylene and butadiene, with large differences in the polymeric products depending on the both the choice of the lanthanide and the feed ratio of the monomer. With both neodymium and dysprosium multiblock polymers consisting of 1,4-cis, 1,4-trans, and ethylene blocks were obtained. With scandium two distinct modes of polymerization were detected, with both multiblock 1,4-cis and a somewhat alternating ethylene/1,4-trans block structure observed in the 13C NMR spectra.

A New Laboratory Polymerizing Apparatus Equipped with Halogen and Metal Halide Light Sources

We developed a new laboratory polymerizing unit (Twinkle MIII: M3) equipped with different light sources (a metal halide and two halogen lamps) that could illuminate independently. This study introduced the structure of the curing unit and examined the curing depth of a representative indirect composite (Artglass). The composite material was cured with the following modes: 1) M3 - halogen only; 2) M3 - metal halide only; 3) M3 - both halogen and metal halide (HM); 4) conventional xenon; and 5) preliminary halogen. The results obtained from analysis of variances and post-hoc tests revealed that curing depth was affected significantly (p < 0.05) by both exposure mode and time, and that HM mode exhibited the most favorable curing performance (p < 0.05) among the curing units and polymerization modes. In conclusion, the M3 unit was found to be well suited to polymerizing the composite material tested.

Dynamic changes in the length distribution of actin filaments during polymerization can be modulated by barbed end capping proteins

During actin polymerization, it has been theorized that the actin filament length distribution initially grows in the form of a Gaussian before converting to produce that of an exponential. However, it has been difficult to demonstrate this experimentally. In this study, we use modern fluorescence microscopy techniques to observe the changing actin filament length distribution during and subsequent to the polymerization process. Nucleated actin filament growth using barbed end capping proteins (gelsolin and erythrocyte capping protein) leads to Gaussian length distributions that are relatively stable. As predicted, nucleated actin filament growth using actin/spectrin complexes follows a similar process until polymerization reaches equilibrium whereafter the Gaussian length distribution rapidly converts to that of an exponential. This study provides direct confirmation of the original theories for the mode of actin polymerization but raises doubts regarding the mechanism of the length distribution conversion from Gaussian to exponential.

Lessons from Simulation Regarding the Control of Synthetic Self-Assembly

AbstractWe investigate the role of particle potential symmetry on self-assembly by Monte Carlo simulation with the particular view towards synthetically creating structures of prescribed form and function. First, we establish a general tendency for the rotational potential symmetries of the particles to be locally preserved upon self-assembly. Specifically, we find that a dipolar particle potential, having a continuous rotational symmetry about the dipolar axis, gives rise to chain formation, while particles with multipolar potentials (e.g., square quadrupole) having discrete rotational symmetries led to the self-assembly of random surface polymers preserving the rotational symmetries of the particles within these sheet structures. Surprisingly, these changes in self-assembly geometry with the particle potential symmetry are also accompanied by significant changes in the thermodynamic character and in the kinetics of the self-assembly process. Linear chain growth involves a continuous chain growth process in which the chains break and reform readily, while the growth of the two-dimensional polymers only occurs after an ‘initiation’ or ‘nucleation’ time that fluctuates from run to run. We show that the introduction of artificial seeds provides an effective method for controlling the structure and growth kinetics of sheet-like polymers. The significance of these distinct modes of polymerization on the functional character of self-assembly growth is illustrated by constructing an artificial centrosome structure derived from particles having continuous and discrete rotational potential symmetries.

Binary frontal polymerization: Velocity dependence on initial composition

Abstract Frontal polymerization is a mode of polymerization in which a localized zone of reaction propagates through the coupling of thermal diffusion and the Arrhenius dependence of the reaction rate. The dependence of the front propagation velocity on the initial composition has been determined in initially miscible binary systems of a free-radically cured diacrylate and an amine- or cationically cured epoxy resin. A minimum of the velocity as a function of the monomer mole fraction is observed if the two polymerizations occur independently. Excellent agreement with an analytical description was found with the diacrylate and an amine-cured epoxy but not for a diacrylate and a cationically cured one because of the effect of HCl impurities on the peroxide.

The effect of pulse activation on microleakage of a ‘packable’ composite resin and two ‘ormocers’

The purpose of the study was to examine the effect of pulse polymerization on microleakage of one packable composite resin and two organically modified ceramics (ormocers), within a high C-factor preparation. Class-V cavity preparations with occlusal margins in enamel and gingival margins in dentin were prepared on 60 freshly extracted premolars or molars. Teeth were randomly assigned to 12 experimental groups (five teeth each) representing the different restorative systems, mode of polymerization and thermal cycling test. Three groups of 20 teeth each were restored with one of the following restorative systems: Scotch bond 1/Filtek P 60, Admira bond/Admira, Etch&Prime 3.0/Definite. 10 restorations of each restorative system (two experimental groups) were conventionally polymerized, whereas the other 10 restorations were polymerized with a pulse program. The specimens were stored in double-distilled water at 37 degrees C for 24 h. Ten restorations from each restorative system (30 restorations, half of the specimens) were thermocycled, the other 30 restorations were kept in double-distilled water at 37 degrees C for 24 h. Then all 60 restorations stained with dye, sectioned and scored for microleakage. Results showed that there were no significant differences (P > 0.05) among the materials with respect to microleakage scores. The experimental groups that were conventionally polymerized revealed significantly more microleakage (P < 0.05) after thermal cycling than all the other groups. The gingival microleakage was significantly more (P < 0.05) than occlusal in conventionally polymerized, thermocycled experimental groups.

Influence of polymerization mode of dual-polymerized resin direct core foundation systems on bond strengths to bovine dentin

Little is known about how various polymerization combinations of dual-polymerized adhesives and resin core foundation pastes affect dentin bond strength. This study examined the influence of different polymerization modes of dual-polymerized resin direct core foundation systems on dentin bond strength using bovine dentin. Two commercially available dual-polymerized resin direct core foundation systems, Clearfil DC Core and UniFil Core, were studied. Bovine mandibular incisors were mounted in autopolymerizing resin and the facial dentin surfaces were ground wet on 600-grit SiC paper. Dentin surfaces were treated according to manufacturer's recommendations, and dual-polymerized adhesives were applied and dual-polymerized (control) or only chemically polymerized. The core foundation resins were condensed into the mold (4 x 2-mm) and were also either dual-polymerized or only chemically polymerized. Eighty specimens (n=10) were stored in 37 degrees C water for 24 hours, then shear tested at a crosshead speed of 1.0 mm/min in a universal testing machine. Fourier transformation infrared spectroscopy was used to measure the degree of conversion of dual-polymerized resin core foundation pastes with both modes of polymerization. One- and 2-way ANOVA, Tukey HSD tests, and Student t -tests were performed (alpha=.05). The highest mean bond strengths (+/-SD) were obtained when the adhesives and resin pastes were dual-polymerized for both core foundation systems (14.6 +/- 2.1 MPa for Clearfil DC Core, 15.6 +/- 2.2 MPa for UniFil Core). Chemical polymerization of dual-polymerized adhesives and resin pastes resulted in significantly lower bond strength (2.8 +/- 0.6 MPa for Clearfil DC Core, 8.1 +/- 2.2 MPa for UniFil Core) ( P <.001). The degree of conversion for the resin pastes decreased significantly ( P <.05) when only chemical polymerization was employed, from 81.5% to 76.1% for Clearfil DC Core and 86.7% to 81.2% for UniFil Core. The data suggests that the dentin bond strengths of the dual-polymerized resin direct core foundation systems are highest when both adhesive and resin core paste are dual polymerized.

In vitro shear bond strength of cementing agents to fixed prosthodontic restorative materials

Durable bonding to fixed prosthodontic restorations is desirable; however, little information is available on the strength of the bond between different cements and fixed prosthodontic restorative materials. This study determined the shear-bond strength of cementing agents to high-gold-content alloy castings and different dental ceramics: high-strength aluminum oxide (Procera AllCeram), leucite-reinforced (IPS Empress), and lithium disilicate glass-ceramic (IPS Empress 2). Prepolymerized resin composite cylinders (5.5 mm internal diameter, n=20) were bonded to the pretreated surfaces of prosthodontic materials. High-gold-content alloy and high-strength aluminum oxide surfaces were airborne-particle-abraded, and pressable ceramics were hydrofluoric acid-etched and silanized prior to cementing. The cementing agents tested were a zinc-phosphate cement (Fleck's zinc cement), glass ionomer cements (Fuji I, Ketac-Cem), resin-modified glass ionomer cements (Fuji Plus, Fuji Cem, RelyX Luting), resin cements (RelyX ARC, Panavia F, Variolink II, Compolute), and a self-adhesive universal resin cement (RelyX Unicem). Half the specimens (n=10) were tested after 30 minutes; the other half (n=10) were stored in distilled water at 37 degrees C for 14 days and then thermal cycled 1000 times between 5 degrees C and 55 degrees C prior to testing. Shear-bond strength tests were performed using a universal testing machine at a constant crosshead speed of 0.5 mm/min. Statistical analysis was performed by multifactorial analysis of variance taking interactions between effects into account. For multiple paired comparisons, the Tukey method was used (alpha=.05). In a 3-way ANOVA model, the main factors substrate, cement, time, and all corresponding interactions were statistically significant (all P <.0001). In subsequent separate 1-way or 2-way ANOVA models for each substrate type, significant differences between cement types and polymerizing modes were found (all P <.001). None of the cement types provided the highest bonding values with all substrate types. After 14 days of water storage followed by thermal cycling, only the self-adhesive universal resin cement (RelyX Unicem) and 2 of the resin cements (Panavia F and Compolute) exhibited strong bond strengths to specific prosthodontic materials. In contrast, zinc-phosphate, glass ionomer, and resin-modified glass ionomer cements showed the lowest values of all tested cementing agents after 14 days of water storage followed by thermal cycling.

DnaG interacts with a linker region that joins the N- and C-domains of DnaB and induces the formation of 3-fold symmetric rings.

Loading of the replicative ring helicase onto the origin of replication (oriC) is the final outcome of a well coordinated series of events that collectively constitute a primosomal cascade. Once the ring helicase is loaded, it recruits the primase and signals the switch to the polymerization mode. The transient nature of the helicase-primase (DnaB-DnaG) interaction in the Escherichia coli system has hindered our efforts to elucidate its structure and function. Taking advantage of the stable DnaB-DnaG complex in Bacillus stearothermophilus, we have reviewed conflicting mutagenic data from other bacterial systems and shown that DnaG interacts with the flexible linker that connects the N- and C-terminal domains of DnaB. Furthermore, atomic force microscopy (AFM) imaging experiments show that binding of the primase to the helicase induces predominantly a 3-fold symmetric morphology to the hexameric ring. Overall, three DnaG molecules appear to interact with the hexameric ring helicase but a small number of complexes with two and even one DnaG molecule bound to DnaB were also detected. The structural/functional significance of these data is discussed and a speculative structural model for this complex is suggested.

Acrylamide in French fries: influence of free amino acids and sugars.

The free amino acid profile and sugar (fructose, glucose, and sucrose) composition were determined in potato samples selected to give a large range of variation (a total of 66 samples). From these samples French fries were produced in a laboratory-scale simulation of an industrial process followed by a finish fry at 180 degrees C for 3.5 min using a restaurant fryer. The final product was blast frozen and analyzed for acrylamide. Acrylamide was detected in all samples, but its concentration varied significantly from 50 to 1800 ng/g. For isotope dilution (13C3) acrylamide analysis, samples were extracted with water, cleaned up on HLB Oasis polymeric and Accucat mixed mode anion and cation exchange SPE columns, and analyzed by LC-MS/MS. Statistical analysis of the data indicates that the effect of sugars and asparagine on the concentration of acrylamide in French fries is positive and significant (p < 0.001). It appears that one of the ways acrylamide formation in French fries can be effectively controlled is by the use of raw products with low sugar (and to a lesser degree, asparagine) content.

Catena-Poly[[bis(pyridine-κN)cobalt(II)]-μ-oxalato] 0.25-hydrate

The structure of the title compound, {[Co(C2O4)(C5H5N)2]·0.25H2O}n, has been determined at ca 110 K with relatively high precision, revealing the polymeric aggregation mode of this species.

Detection in situ and characterization of lignin in the G-layer of tension wood fibres of Populus deltoides.

The occurrence of lignin in the additional gelatinous (G-) layer that differentiates in the secondary wall of hardwoods during tension wood formation has long been debated. In the present work, the ultrastructural distribution of lignin in the cell walls of normal and tension wood fibres from poplar (Populus deltoides Bartr. ex Marshall) was investigated by transmission electron microscopy using cryo-fixation-freeze-substitution in association with immunogold probes directed against typical structural motifs of lignin. The specificity of the immunological probes for condensed and non-condensed guaiacyl and syringyl interunit linkages of lignin, and their high sensitivity, allowed detection of lignin epitopes of definite chemical structures in the G-layer of tension wood fibres. Semi-quantitative distribution of the corresponding epitopes revealed the abundance of syringyl units in the G-layer. Predominating non-condensed lignin sub-structures appeared to be embedded in the crystalline cellulose matrix prevailing in the G-layer. The endwise mode of polymerization that is known to lead to these types of lignin structures appears consistent with such an organized cellulose environment. Immunochemical labelling provides the first visualization in planta of lignin structures within the G-layer of tension wood. The patterns of distribution of syringyl epitopes indicate that syringyl lignin is deposited more intensely in the later phase of fibre secondary wall assembly. The data also illustrate that syringyl lignin synthesis in tension wood fibres is under specific spatial and temporal regulation targeted differentially throughout cell wall layers.

Structure-based mutational analysis of the 4'-phosphopantetheinyl transferases Sfp from Bacillus subtilis: carrier protein recognition and reaction mechanism.

The activation of apo-peptidyl carrier proteins (PCPs) of nonribosomal peptide synthetases (NRPSs), apo-acyl carrier proteins (ACPs) of polyketide synthases (PKSs), and fatty acid synthases (FASs) to their active holo form is accomplished with dedicated 4'-phosphopantetheinyl transferases (PPTases). They catalyze the transfer of the essential prosthetic group 4'-phosphopantetheine (4'-Ppant) from coenzyme A (CoA) to a highly conserved serine residue in all PCPs and ACPs. PPTases, based on sequence and substrate specifity, have been classified into three types: bacterial holo-acyl carrier protein synthase (AcpS), fatty acid synthase of eukaryotes (FAS2) and Sfp, a PPTase of secondary metabolism. The recently solved crystal structures of AcpS and Sfp-type PPTases with CoA revealed a common alpha + beta-fold with a beta(1)alpha(3)beta(2) motif and similarities in CoA binding and polymerization mode. However, it was not possible to discern neither the PCP binding region of Sfp nor the priming reaction mechanism from the Sfp-CoA cocrystal. In this work, we provide a model for the reaction mechanism based on mutational analysis of Sfp that suggests a reaction mechanism in which the highly conserved E151 deprotonates the hydroxyl group of the invariant serine of PCP. That, in turn, acts as a nucleophile to attack the beta-phosphate of CoA. The Sfp mutants K112, E117, and K120 further revealed that the loop region between beta4 and alpha5 (residues T111-S124) in Sfp is the PCP binding region. Also, residues T44, K75, S89, H90, D107, E109, E151, and K155 that have been shown in the Sfp-CoA cocrystal structure to coordinate CoA are now all confirmed by mutational and biochemical analysis.

Flexural strength and modulus of a step‐cured resin composite

In an attempt to reduce marginal contraction gaps, a step‐curing mode of light polymerization of resin composite has been proposed. It was hypothesized that such an exposure mode, having an initial reduced curing rate, would result in composite having a lower modulus of elasticity than composite exposed in one step. The composite was initially exposed to power density levels of 50, 100, 150, or 200 mW/cm2for durations of 10, 20, or 40 s. The final exposure and the exposure of the control group were performed at 750 mW/cm2for 20 s. It was found that certain modes of two‐step exposure resulted in a lower flexural modulus than did a one‐step exposure of constant, high power density. Regression analysis showed, with statistical significance, that flexural modulus was relatively high following a short initial exposure at low power density or using a long initial exposure at high power density. Conversely, flexural modulus was relatively low following a long initial exposure at low power density or using a short initial exposure at high power density. The results were explained by probable differences in degree of conversion and cross‐link density of the polymer network.

Influence of adhesive polymerization mode on dentin bond strength of direct core foundation systems

This study examined the influence of various adhesive systems on dentin bond strength of direct core foundation resins. Two commercially available direct core foundation resin systems and 2 adhesive polymerization modes were used. Facial bovine dentin surfaces were wet ground on 600-grit SiC paper. Dentin surfaces were treated according to the manufacturers' instructions and were light polymerized (control). Chemical- and light-polymerized adhesive systems were used separately. The resin paste was condensed into a mold and bonded to the dentin surface. Ten specimens per test group were stored in water at 37 degrees C for 24 hours, and a shear test was conducted at a crosshead speed of 1.0 mm/minute using a universal testing machine. Analysis of variance (ANOVA) and Duncan's multiple comparison test were performed (alpha = 0.05). Dual polymerization of resin pastes revealed higher bond strength with the combination of light-polymerized adhesive (22.8-24.3 MPa), but significantly lower bond strength with the combination of a chemical-polymerized adhesive (4.2-5.7 MPa). The present data suggests that dentin bond strengths in direct core foundation systems can be influenced by the combination of adhesive and resin paste.

Thermodynamic Dissection of the Polymerizing and Editing Modes of a DNA Polymerase

DNA polymerases with intrinsic proofreading activity interact with DNA primer/templates in two distinct modes, corresponding to the complexes formed during the 5′–3′ polymerization or 3′–5′ editing of a nascent DNA chain. Thermodynamic measurements designed to quantify the energetic contributions of individual DNA–protein contacts in either the polymerizing or editing complexes are complicated by the fact that both species exist in solution and are not resolved in conventional DNA–protein binding assays. To overcome this problem, we have developed a new binding analysis that combines information from steady-state and time-resolved fluorescence experiments and uses the Klenow fragment of Escherichia coli DNA polymerase I (KF) and fluorescently labeled primer/template oligonucleotides as a model polymerase–DNA system. Steady-state fluorescence titrations are used to evaluate the overall affinity of KF for the primer/template, while time-resolved fluorescence anisotropy is used to quantify the equilibrium fractions of the primer/template bound in the polymerizing and editing modes. From a combined analysis of both data, the equilibrium constant and hence standard free energy change associated with each binding mode can be obtained unequivocally. This method is initially used to determine the equilibrium constants describing binding of a correctly base-paired primer/template to the 5′–3′ polymerase and 3′–5′ exonuclease sites of KF. It is then extended to quantify the extent to which these parameters are affected by the introduction of mismatches into the primer/template, and by rearrangement of specific side-chains in the exonuclease domain of the protein. While these perturbants were originally designed to demonstrate the utility of our new approach, they are also relevant in their own right since they have helped identify some hitherto unknown determinants of polymerase fidelity.

Heat-Resistant Organic Matrices Quo Vadis? (Review)

The introduction is a brief review of the characteristics desired for heat-resistant composite matrices, and the two possible approaches based on linear polymers and telechelic oligomers that can be cross-linked during use. Special attention is paid to cross-linkable oligomers, whose use currently poses fewer problems. The different structures in question, i.e. bismaleimides, bismaleimides (modified by cyanates or epoxies), bisnadimides, acetylene and propargylic compounds, are discussed in terms of chemistry (stability of oligomers, mode of polymerization, aging) and mechanical properties (ductility). Several solutions little used at the present time (semi-interpenetrated networks, molecular composites, polymerizable molecular assemblies, hyper branched oligomers and liquid crystal monomers, etc.) are discussed in terms of the future. There may be technical solutions for use up to 300°C, but industrial development is far from being resolved because of, among other things, the small market for these products.

A Novel Mode of Polymerization of α1-Proteinase Inhibitor

Patients homozygous for the Z mutant form of alpha1-proteinase inhibitor (alpha1-PI) have an increased risk for the development of liver disease because of the accumulation in hepatocytes of inclusion bodies containing linear polymers of mutant alpha1-PI. The most widely accepted model of polymerization proposes that a linear, head-to-tail polymer forms by sequential insertion of the reactive center loop (RCL) of one alpha1-PI monomer between the central strands of the A beta-sheet of an adjacent monomer. This model derives primarily from two observations: peptides that are homologous with the RCL insert into the A beta-sheet of alpha1-PI monomer and this insertion prevents alpha1-PI polymerization. Normal alpha1-PI monomer does not spontaneously polymerize; however, here we show that the disulfide-linked dimer of normal alpha1-PI spontaneously forms linear polymers in buffer. The monomers within this dimer are joined head-to-head. Thus, the arrangement of monomers in these polymers must be different from that predicted by the loop-A sheet model. Therefore, we propose a new model for alpha1-PI polymer. In addition, polymerization of disulfide-linked dimer is not inhibited by the presence of the peptide even though dimer appears to interact with the peptide. Thus, RCL insertion into A beta-sheets may not occur during polymerization of this dimer.