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Actin polymerization induced by GTP gamma S in permeabilized neutrophils is induced and maintained by free barbed ends.

To address the mechanisms through which agonists stimulate actin polymerization, we examined the roles of monomer sequestering proteins and free barbed ends on actin polymerization induced by guanosine 5'-3-O-(thio)triphosphate (GTP gamma S) in neutrophils permeabilized with streptolysin O. Addition of profilin (without GTP gamma S) caused a net decrease in F-actin. Thus, merely making profilin available in the cell was not sufficient to induce actin polymerization. On the other hand, addition of profilin hardly affected the polymerization induced by GTP gamma S, while thymosin beta 4 or DNase I decreased this polymerization. These data suggested that GTP gamma S induced polymerization by increasing the availability of barbed ends. In the presence of cytochalasin B, profilin did inhibit polymerization induced by GTP gamma S, demonstrating that GTP gamma S did not inhibit profilin's monomer sequestering ability. The F-actin induced by GTP gamma S was not limited by a time-dependent loss of G-actin or G-proteins from permeabilized cells since, following stimulation with suboptimal concentrations of GTP gamma S, addition of more GTP gamma S induced further polymerization. Barbed ends remained free after F-actin reached plateau since (a) cytochalasin B caused depolymerization of induced F-actin and (b) profilin did not depolymerize induced F-actin unless the cells were first treated with cytochalasin to cap barbed ends. The data indicate that GTP gamma S maintains an increased level of F-actin by keeping at least a few barbed ends available for polymerization.

Actins↗

Bundling of actin filaments by elongation factor 1 alpha inhibits polymerization at filament ends.

Elongation factor 1 alpha (EF1 alpha) is an abundant protein that binds aminoacyl-tRNA and ribosomes in a GTP-dependent manner. EF1 alpha also interacts with the cytoskeleton by binding and bundling actin filaments and microtubules. In this report, the effect of purified EF1 alpha on actin polymerization and depolymerization is examined. At molar ratios present in the cytosol, EF1 alpha significantly blocks both polymerization and depolymerization of actin filaments and increases the final extent of actin polymer, while at high molar ratios to actin, EF1 alpha nucleates actin polymerization. Although EF1 alpha binds actin monomer, this monomer-binding activity does not explain the effects of EF1 alpha on actin polymerization at physiological molar ratios. The mechanism for the inhibition of polymerization is related to the actin-bundling activity of EF1 alpha. Both ends of the actin filament are inhibited for polymerization and both bundling and the inhibition of actin polymerization are affected by pH within the same physiological range; at high pH both bundling and the inhibition of actin polymerization are reduced. Additionally, it is seen that the binding of aminoacyl-tRNA to EF1 alpha releases EF1 alpha's inhibiting effect on actin polymerization. These data demonstrate that EF1 alpha can alter the assembly of F-actin, a filamentous scaffold on which non-membrane-associated protein translation may be occurring in vivo.

Actin Cytoskeleton↗

Free and polymerized tubulin in cultured bone cells and Chinese hamster ovary cells: the influence of cold and hormones.

Free and polymerized tubulin were measured in bone cells and Chinese hamster ovary (CHO) cells cultured on plastic substrata. Polymerized tubulin was stabilized in a microtubule- stabilizing medium (MSM) containing 50 percent glycerol and separated from free tubulin by centrifugation. Tubulin content was assayed in both fractions by the colchicines- binding assay. The measured degree of polymerization in both bone cells and CHO cells varied with stabilixation conditions. The degree of polymerization in both bone cells and CHO cells varied with stabilization conditions. The degree of polymerization in both bone cells and CHO cells varied with stabilization conditions. The degree of polymerization in attached cells was found to increase up to 73 percent during the first 20 min after addition of the MSM at 24 degrees C, and remained constant thereafter. Stabilization of 0 degrees C resulted in a decrease down to 62 percent in the degree of constant thereafter. Stabilization at 0 degrees C resulted in a decrease down to 62 percent in the degree of polymerization during the first 20 min after addition of the MSM at 24 degrees C, and remained constant thereafter. Confluent bone cells maintained at 0 degrees C for 1 h before stabilization contained significantly less polymerized tubulin than control cells kept at 37 degrees C using stabilization both at 0 degrees C and at 24 degrees C. Changes in bone cell morphology induced by incubation of cells with prostaglandin E(1) or E(2), parthyroid hormone, and dibutyryl cyclic AMP were not associated with a change in the degree of tubulin polymerization. This was confirmed morphologically by immunofluorescence using affinity-purified tubulin antibodies: microtubules in hormone- treated cells were not noticeably reorganized when compared to microtubule organization in control cells. They were, however, squeezed closer together in cellular pseudopods due to the altered cell shape. This altered cell shape appears to be correlated with disorganization of the microfilament system, since microfilaments, detected using affinity-purified actin antibodies, did alter drastically their appearance and distribution after hormone addition.

Alprostadil↗

Equilibrium polymerization in the Stockmayer fluid as a model of supermolecular self-organization.

A diverse range of molecular self-organization processes arises from a competition between directional and isotropic van der Waals intermolecular interactions. We conduct Monte Carlo simulations of the Stockmayer fluid (SF) with a large dipolar interaction as a minimal self-organization model and focus on basic thermodynamic properties that are needed to characterize the polymerization transition that occurs in this fluid. In particular, we determine the polymerization transition lines from the maximum in the specific heat, C(v), and the inflection point in the extent of polymerization, Phi. We also characterize the geometry (radius of gyration R(g), chain length L, chain topology) of the clusters that form in this associating fluid as a function of temperature, T, and concentration, rho . The pressure, P, and the second virial coefficient, B2, were determined, since these properties contain essential information about the strength of the isotropic (van der Waals) interactions. Our simulations indicate that the locations of the polymerization lines are quantitatively consistent with a model of equilibrium polymerization with the enthalpy of polymerization ("sticking energy") fixed by the minimum in the intermolecular potential. The polymerization transition in the SF is accompanied by a topological transition from predominantly linear to ring polymers upon cooling that is driven by the minimization of the dipolar energy of the clusters. We also find that the basic interaction parameters describing polymerization and phase separation in the SF can be estimated based on the existing theory of equilibrium polymerization, but the theory must be refined to account for ring formation in order to accurately describe the configurational properties of this model self-organizing fluid.

Journal Article↗

Cysteine-independent polymerization of metallothioneins in solutions and in crystals.

Polymerization of metallothioneins is one of the usually encountered puzzles during the research process of metallothioneins' structure and function. Our work focuses on the cysteine independently occurred polymerization from metallothioneins monomers in different milieus, while it leaves out the aggregation caused by the oxidation of cysteine, because the latter circumstance is the result of purification lapsus. After the purification of metallothioneins monomers, a dynamic light-scattering technique is used to detect the polymerized states of rabbit liver metallothionein I and II in different buffers, which is the first systematical detection of polymerized states of metallothioneins in solutions. The effects of different compositions of each buffer are discussed in details. Steric complementarity, hydrophobic, and electrostatic interaction characteristics are studied, following the modeling of monomers and relevant polymers of rat metallothionein II, rabbit liver metallothionein I and II. These theoretical calculations are the first complete computer simulations on different factors affecting metallothioneins' polymerization. A molecular recognition mechanism of metallothioneins' polymerization in solutions is proposed on the bases of experimental results and theoretical calculations. Preliminary X-ray studies of two crystal forms of rabbit liver metallothionein II are compared with the crystal structure of rat metallothionein II, and the polymerized states in crystal packing are discussed with the knowledge of polymerization of metallothioneins in solutions. The hypothesis, which is consistent with theoretical calculations and experimental results, is expected to construct a connection between the biochemical characteristics and physiological functions of metallothioneins, and this research may give some enlightenment to the topics of protein polymerizations.

Amino Acid Sequence↗

Growth hormone promotion of tubulin polymerization stabilizes the microtubule network and protects against colchicine-induced apoptosis.

We have investigated the effect of GH on microtubular physiology in Chinese hamster ovary (CHO) cells stably transfected with the complementary DNA for the rat GH receptor (CHO-GHR(1-638)). We show here that after 30 min of human GH (hGH) treatment of CHO-GHR(1-638) cells, there was a significant increase in the level of polymerization of all four tubulin isoforms (alpha-, beta-, gamma-, and tyrosinated alpha-tubulin) compared with the serum-deprived state. However, this transient increase in the levels of polymerized tubulin after hGH treatment was particularly pronounced for beta- and tyr alpha-tubulin. For alpha- and gamma-tubulin, the hGH-induced increase in polymerization state lasted to approximately 3 h and then declined by 7 h, whereas for beta- and tyr alpha-tubulin there was a decrease in the polymerization state at 1-2 h after hGH treatment compared with the level at 30 min (but still greater than the serum-deprived state) followed by a second but lesser wave of increased polymerization lasting to 7 h. The changes in the polymerization state of the tubulins were not accompanied by comparative changes in the level of total cellular tubulin. The proline rich box 1 region of the GH receptor was required for hGH to stimulate tubulin polymerization indicative that this event is JAK dependent. Increased tubulin polymerization still occurred in response to hGH in a receptor truncation lacking the carboxyl terminal half of the intracellular domain of the GH receptor indicative that hGH induced changes in intracellular calcium concentration is not required for tubulin polymerization. Prior treatment of CHO-GHR(1-638) cells with hGH retarded colchicine induced microtubule depolymerization and also prevented colchicine induced apoptotic cell death. The integrity of the microtubule network was not required for GH-induced STAT5 mediated transcription as treatment of cells with colchicine, vincristine, or vinblastine did not alter the fold stimulation of the STAT5 mediated transcriptional response to GH. Thus one consequence of cellular treatment with GH is alteration in microtubule physiology.

Animals↗

The linear shrinkage and microhardness of packable composites polymerized by QTH or PAC unit.

This study evaluated the effectiveness of a plasma arc curing (PAC) unit for packable resin composite curing. The amount and speed of polymerization shrinkage and the microhardness of packable composites were evaluated in order to compare the PAC unit's effectiveness with a quartz tungsten halogen (QTH) unit. Sure Fil (Dentsply Caulk), Pyramid (BISCO Inc) and Synergy Compact (Colténe/Whaledent) were used as the packable composites. In the case of curing with the PAC unit, the composites were light cured with Apollo 95E (DMD System Inc) for 1 second (Group 1), 2 seconds (Group 2), 3 seconds (Group 3), 6 seconds (Group 4) and 12 seconds (Group 5). For light curing with the QTH unit, the composites were light cured for 60 seconds using XL3000 (Group 6). The linear polymerization shrinkage of each composite was measured using a custom made linometer, and the data was stored in a computer every 0.5 to 0.55 seconds for a total of 60 seconds. For each composite, the amount of polymerization was compared using one-way ANOVA with Tukey at the 95% confidence level. In order to compare the speed of polymerization, the peak time (PT), showing the highest speed of polymerization and maximum speed of polymerization (Smax), were determined from the data and compared using one-way ANOVA with Tukey at the 95% confidence level for each material. Based on the statistical analysis among the PAC-cure groups (Groups 1 through 5), the group that was not statistically different from the QTH-cure group (Group 6) in the amount of linear polymerization shrinkage was determined for each material, and the corresponding curing time of the group was defined as the tentative minimum PAC-curing time (TMPT). For microhardness measurements, the samples were placed in a 2-mm thick Teflon plate. Twenty specimens, randomly divided into the PAC-cure group (Group 1) or the QTH-cure group (Group 2), were prepared for each material. In Group 1, each composite was light cured for TMPT with the PAC unit. In Group 2, each composite was light cured for 60 seconds with the QTH unit. Microhardness was measured on the upper and lower surface. For each material, the microhardness of the upper and lower surface of Groups 1 and 2 was analyzed using two-way ANOVA with Tukey at the 95% confidence level. The amount of polymerization was Group 1<Group 2<Group 3<Group 4<Groups 5, 6 in the Sure Fil composite (p<0.05); Groups 1, 2<Group 3<Groups 4, 6<Group 5 in the Synergy Compact composite (p<0.05) and Group 1 <Group 2 <Group 3 <Groups 4, 6 <Group 5 in the Pyramid composite (p<0.05) Regarding the speed of polymerization, the order of PT was G1, G2, G3<G4, G5<G6 (p<0.05). The order of Rmax was G6<G1, G2, G3, G4, G5 in Sure Fil; G6<G1<G2, G3, G4<G5 in Synergy Compact (p<0.05) and G6<G1<G2, G3<G4, G5 in Pyramid (p<0.05) On the upper surface, there was no statistical difference in microhardness between Groups 1 and 2 in all materials. On the lower surface, the microhardness of Group 2 was significantly higher than Group 1 in all materials. In all materials in Group 1 and the Synergy Compact of Group 2, microhardness of the upper surface was significantly higher than the lower surface (p<0.05). In Sure Fil and Pyramid of Group 2, there was no difference in microhardness between the upper and lower surfaces.

Composite Resins↗

Some mechanical properties of a highly cross-linked, microwave-polymerized, injection-molded denture base polymer.

PURPOSE: The impact strength and the flexural properties of denture base materials are of importance in predicting their clinical performance upon sudden loading. This study compares the impact and transverse strengths and the flexural modulus of three denture base polymers. MATERIALS AND METHODS: The investigation included a relatively new microwave-polymerized polyurethane-based denture material processed by an injection-molding technique, a conventional microwave-polymerized denture material, and a heat-polymerized compression-molded poly(methyl methacrylate) (PMMA) denture material. Impact strength was determined using a Charpy-type impact tester. The transverse strength and the flexural modulus were assessed with a three-point bending test. The results were subjected to statistical analysis using a one-way analysis of variance and the Scheffé test for comparison. RESULTS: The impact strength of the microwave-polymerized injection-molded polymer was 6.3 kl/m2, while its flexural strength was 66.2 MPa. These values were lower than those shown by the two compression-molded PMMA-based polymers. The differences were statistically significant. The flexural modulus of the new denture material was 2,832 MPa, which was higher than the conventional heat-polymerized polymer but was comparable to the other microwave-polymerized PMMA-based polymer. The difference in the flexural modulus was statistically significant. CONCLUSION: In terms of the impact and flexural strengths, the new microwave-polymerized, injection-molded, polyurethane-based polymer offered no advantage over the existing heat- and microwave-polymerized PMMA-based denture base polymers. However, it has a rigidity comparable to that of the microwave-polymerized PMMA polymer.

Acrylic Resins↗

Post-gel polymerization contraction of "low shrinkage" composite restoratives.

This study compared the post-gel contraction of two "low-shrinkage" composites (InTen-S [IS], Ivoclar-Vivadent; Aelite LS [AL], BISCO Inc) and an ormocer (Admira [AM], Voco) to two conventional mini-filled composites (Renew [RN], BISCO; Z100 [ZO], 3M ESPE). A strain-monitoring device and test configuration were used to measure the linear polymerization shrinkage associated with the various composites (A2 shade) during and up to 60 minutes post light polymerization. Each specimen was irradiated for 40 seconds using a halogen curing light (Max, Dentsply-Caulk) with an intensity of 401 mW/cm2. Five specimens were made for each composite. Data was analyzed using one-way ANOVA/Scheffe's post-hoc test at significance level 0.05. The linear percentage shrinkage immediately after light polymerization and at 60 minutes post light polymerization ranged from 0.10 +/- 0.02 to 0.40 +/- 0.02% and 0.22 +/- 0.02 to 0.60 +/- 0.05%, respectively. Post-gel shrinkage ranking of the materials was as follows: immediately after light polymerization - IS < AL < AM < ZO < RN and at 60 minutes post light polymerization - IS < AL = AM < ZO < RN. The shrinkage associated with IS, AL and AM was significantly lower than for ZO and RN immediately after light polymerization and at 1, 10, 30 and 60 minutes post light polymerization. The post-gel polymerization shrinkage of IS, AL and AM was significantly lower than conventional mini-filled composites.

Analysis of Variance↗

Effectiveness and ergonomic features of different light sources for polymerization of composite resin materials.

INTRODUCTION: Besides the halogen curing lights, already accepted as standard equipment, diode and plasma-arc units for polymerization of filling materials have also been commercially available in Bulgaria in recent years. The present study was prompted by the lack of detailed scientific information about these novel light curing devices. AIM: 1. To evaluate the quality of polymerization achieved by different light curing units by measuring the quantity of residual monomer in the test specimens. 2. To classify the units according to their convenience features and polymerization abilities. MATERIALS AND METHODS: Five diode units from three different manufacturers, one plasma-arc unit and one halogen unit were used. The test specimens, made from the hybrid photopolymerizing material Illumine (Ortho Plus, France), Type II, radiopaque, color C1, were 2 mm high and 4 mm in diameter. They were cured for 40 seconds with the respective unit after which they were ground and 25 mg from each sample were immersed in 8 ml of 95% ethanol for extraction of the residual monomer. The quantity of the residual monomer was determined by the method of the standard straight line. Statistical analysis was performed with the analysis of variance and F-test. The comparisons in each group were made by the Tukey-Kramer test, the level of significance for the null hypothesis being P = 0.05. The complex evaluation of the units was made with the help of a scale, developed by us. RESULTS: The material polymerized by the halogen light demonstrated the smallest quantities of residual monomer of all the specimens. There was no statistically significant difference between the quantities of the residual monomer in the specimens polymerized by the plasma arc unit (PAC), OL2 and Elipar Free Light exp. Such a difference, however, existed when comparing the PAC unit and the 3M ESPE unit, working in a continuous irradiation mode, the former having more residual monomer. Polymerization by Elipar Free Light produced the smallest quantities of residual monomer of all other diode lights. The only exception was the comparison between OL2 and Elipar Free Light exp., where there was no difference. The diode light of 3M ESPE performed best of all units according to the criteria we used for evaluation of the light curing devices. CONCLUSIONS: 1. The intensity of radiation of the light source and the duration of polymerization are of crucial importance for the complete conversion of the monomer into polymer. 2. Diode technology has not yet been able to reach any halogen equipment with respect to quality of polymerization, despite some of its unquestionable advantages. 3. When choosing a light curing unit, the quality of polymerization is more important than the price and the offered additional features.

Composite Resins↗

The polymerization of acetyl-CoA carboxylase.

Citrate, an allosteric activator of acetyl-CoA carboxylase, induces polymerization of an inactive protomeric form of the enzyme into an active filamentous form composed of 10-20 protomers. The light-scattering properties of the carboxylase were used to study the kinetics of its polymerization and depolymerization. From stopped flow kinetic studies, we have established that polymerization is a second order process, with a second order rate constant of 597,000 M-1 s-1. There appear to be two steps which limit polymerization of the inactive carboxylase protomer: 1) a rapid citrate-induced conformational change which is independent of enzyme concentration and leads to an active protomeric form of the enzyme (Beaty, N. B., and Lane, M. D. (1983) J. Biol. Chem. 258, 13043-13050, preceding paper) and 2) the dimerization of the active protomer, which constitutes the first step of polymerization and is enzyme concentration-dependent. Dimerization is the rate-limiting step of acetyl-CoA carboxylase polymerization. Depolymerization of fully polymerized acetyl-CoA carboxylase is caused by malonyl-CoA, ATP X Mg, and Mg2+. Both malonyl-CoA and ATP X Mg (and HCO-3) compete with citrate in the maintenance of a given state of the protomer-polymer equilibrium apparently by carboxylating the enzyme to form enzyme-biotin-CO-2 which destablizes the polymeric form. Free citrate is the species responsible for polymerizing the enzyme and Mg2+ causes depolymerization of the enzyme by lowering the concentration of free citrate.

Acetyl-CoA Carboxylase↗

Nucleation-controlled polymerization of human monoclonal immunoglobulin G cryoglobulins.

The kinetics of the polymerization of human monoclonal cryoimmunoglobulins at low temperature was investigated in temperature jump experiments by monitoring the changes in turbidity resulting from the scattering of incident light by the polymers. Above a critical concentration between 2 and 3 mg/ml, depending on the ionic strength, the kinetics were characterized by a concentration-dependent lag phase and initial rate of self-assembly. Under equilibrium conditions which favored polymerization, the only stable intermediate detected by analytical ultracentrifugation was the dimer. Although purified monomers were unable to self-associate at 4 degrees C, addition of trace amounts of autologous dimers promoted polymerization. The apparent rate of polymerization was shown to be slow (k = 4.7 X 10(-4) M-1 s-1), and the process was governed by an equilibrium constant of 4.6 X 10(4) M-1. The initial rate of self-assembly was proportional to the product of the monomer concentration and the concentration of promoter (i.e. dimer). The rate of depolymerization was three orders of magnitude greater than the rate of polymerization and was proportional to the concentration of polymers present. These results suggest that the polymerization of monoclonal cryoimmunoglobulins is a nucleation-controlled process in which dimerization is the rate-limiting step. Kinetic studies on the polymerization of Fab and F(ab')2 fragments from cryoimmunoglobulins and a comparison of cryogel ultrastructure by electron microscopy suggested that the interaction site between monomers is located in the Fab region. Since the polymerization of monomers was only induced by autologous dimers and not dimers from other cryoimmunoglobulins, it was concluded that the hypervariable regions play a specific role in the condensation reaction. The fact that one cryoimmunoglobulin has a well defined antibody activity against streptolysin O argued against a low temperature-induced auto-anti-idiotype mechanism. Reduction of the interchain disulfide bonds of the Fab fragments abolished their ability to polymerize, probably by inducing a conformational change a considerable distance away in the variable domains of the molecules.

Cryoglobulins↗

Role of Leu-beta 88 in the hydrophobic acceptor pocket for Val-beta 6 during hemoglobin S polymerization.

X-ray crystallographic studies indicate that the hydrophobic acceptor pocket made by E and F helices involving Leu-beta 88 and Phe-beta 85 is critical for the formation of stable hydrophobic interactions with Val-beta 6 on an adjacent deoxy-hemoglobin (Hb) S tetramer. Ala and Phe substitutions at the beta 88 position in Hb S were made using a yeast expression system in an effort to clarify the role of Leu-beta 88 in creating a suitable acceptor site for Val-beta 6 during polymerization of Hb S. Both Ala- and Phe-beta 88 substitutions in Hb S inhibited polymerization compared with Hb S. Critical concentrations for polymerization of alpha 2 beta 2 Val-6,Ala-88 and alpha 2 beta 2Val-6,Phe-88 were 6- and 10-fold higher, respectively, than that of Hb S (alpha 2 beta 2Val-6,Leu-88). Deoxy-Hb S containing Phe-beta 88 polymerized without a delay time like Trp-beta 6- and Phe-beta 6-substituted hemoglobins (Adachi, K., Konitzer, P., Kim, J., Welch, N., and Surrey, S. (1993) J. Biol. Chem. 268, 21650-21656). In contrast, oversaturated deoxy-Hb S containing Ala-beta 88 also polymerized without a delay time; however, with decreasing hemoglobin concentrations, the kinetics of polymerization were biphasic. At lower hemoglobin concentrations, closer to the critical concentration for polymerization, deoxy-Hb S containing Ala-beta 88 polymerized after a distinct delay time. These results suggest that bulky beta 88 hydrophobic replacements like Phe may sterically inhibit insertion of Val-beta 6 into the acceptor pocket. In contrast, smaller sized, less hydrophobic amino acids like Ala compared with Leu-beta 88 may allow insertion of Val-beta 6 into the acceptor pocket but may not promote stable protein-protein interactions with an adjacent Hb molecule. Stereospecificity and hydrophobicity of the Val-beta 6 hydrophobic acceptor pocket as well as the beta 6 amino acid are, therefore, critical for polymerization of deoxy-Hb S.

Biopolymers↗

ESR study of MMA polymerization by a peroxide/amine system: bone cement formation.

Electron spin resonance (ESR) spectroscopy was used to gain insight at the molecular level into the curing of bone cement. Methyl methacrylate was polymerized using a N,N-dimethyl-p-toluidine (TD)/benzoyl peroxide (BPO) redox system in the presence of polymethyl methacrylate (PMMA) powder. The conventional nine-line ESR spectrum for the growing polymer radical was detected at the gel stage of polymerization. While the optimum free radical concentration was observed near the equimolar amine/BPO concentration, excess amine led to a change in the chemical structure of the trapped radical and inhibited the polymerization process. At a high amine/BPO ratio the nine-line signal disappeared and a three-line nitroxide-based radical appeared. The appearance of this nitroxide signal seems to depend on the amine/BPO molar ratio and on the presence of PMMA. An excess amount of amine with respect to BPO was found to inhibit the polymerization process. When BPO was removed, the system still polymerized but with a longer gelation time and a lower radical concentration. These results demonstrate that trapped free radicals in the bulk polymerization of MMA convert to polymeric peroxides that act as initiators in bone cement. When the accelerator 4-dimethylamino phenethyl alcohol (TDOH) was used, a higher radical concentration was observed in the polymerizing system. TDOH shows potential for being a more effective accelerator than TD for bone cement curing.

Amino Alcohols↗

Actin polymerization in human eosinophils, unlike human neutrophils, depends on intracellular calcium mobilization.

Eosinophils represent major effector cells in the allergic inflammation. In contrast to neutrophils, the mechanism of eosinophil activation during the inflammatory response is poorly understood. In this study, the relation between calcium fluxes, chemotaxis, and actin polymerization in eosinophils from healthy non-atopic donors was investigated. Pre-incubation of eosinophils with the intracellular calcium chelator BAPTA dose-dependently prevented an increase in the intracellular calcium concentration ([Ca2+]i), whereas the depletion of extracellular calcium in the test medium had no effect. The chemotactic response of eosinophils, which was measured by the modified boyden chamber technique upon stimulation with RANTES, C5a and PAF, was dose-dependently inhibited by the chelation of intracellular calcium as well as inactivation of the cells in Ca2+ -depleted medium. To evaluate whether other cell functions which are involved in the migratory response of eosinophils might be dependent on intracellular and extracellular calcium, actin polymerization was investigated. Flow-cytometric measurement of F-actin with NBD-phallacidin revealed that actin polymerization in human eosinophils in response to RANTES, C5a, and PAF was dose-dependently inhibited by the intracellular calcium chelator BAPTA. Since it is well known that actin polymerization in neutrophils is not affected by chelation of intracellular calcium, actin polymerization in these cells was investigated under the same conditions as for eosinophils. In contrast to eosinophils, BAPTA did not inhibit actin polymerization in neutrophils. In summary, these data demonstrate that intracellular calcium fluxes represent a prerequisite for eosinophil chemotaxis and actin polymerization in human eosinophils. Furthermore, regulation of actin polymerization in eosinophils differed from that of neutrophils on the level of intracellular calcium fluxes.

Actins↗

Coordination polymerization in water affording amorphous polyethylenes.

The coordination polymerization of ethylene in water as a reaction medium was studied. Rubbery amorphous branched polyethylene was obtained when a known cationic diimine-substituted methyl complex was employed as a catalyst precursor. High rates of up to 900 TOh(-1) (turnover frequency) were observed. In contrast to solution polymerization in an organic solvent, the rate of suspension polymerization in water increases greatly with ethylene pressure in the range up to 20 bar; this indicates control of the polymerization rate by the concentration of the olefin monomer at the catalytically active site. The effect and mode of mass transfer phenomena were studied. A high catalyst stability in the aqueous coordination polymerization was observed. It was found to be due to an "encapsulation" of the water-insoluble catalyst precursor in the hydrophobic amorphous polymer during the polymerization reaction, and this resulted in strongly restricted accessibility for the aqueous phase. Surprisingly, exposure of the water-stable catalyst precursor to ethylene monomer in solution in the presence of water resulted in immediate decomposition. Polymer microstructure, and thermal and mechanical properties were investigated. The different degree of branching, molecular weight, and corresponding macroscopic properties of the polymers obtained in water as a reaction medium versus solution polymerization in methylene chloride under the same conditions are due to the different phase behavior during polymerization (suspension vs. solution), as opposed to an effect of water on the catalytically active centers.

Journal Article↗

Complexity in modeling of residual stresses and strains during polymerization of bone cement: effects of conversion, constraint, heat transfer, and viscoelastic property changes.

Aseptic loosening of cemented joint prostheses remains a significant concern in orthopedic biomaterials. One possible contributor to cement loosening is the development of porosity, residual stresses, and local fracture of the cement that may arise from the in-situ polymerization of the cement. In-situ polymerization of acrylic bone cement is a complex set of interacting processes that involve polymerization reactions, heat generation and transfer, full or partial mechanical constraint, evolution of conversion- and temperature-dependent viscoelastic material properties, and thermal and conversion-driven changes in the density of the cement. Interactions between heat transfer and polymerization can lead to polymerization fronts moving through the material. Density changes during polymerization can, in the presence of mechanical constraint, lead to the development of locally high residual strain energy and residual stresses. This study models the interactions during bone cement polymerization and determines how residual stresses develop in cement and incorporates temperature and conversion-dependent viscoelastic behavior. The results show that the presence of polymerization fronts in bone cement result in locally high residual strain energies. A novel heredity integral approach is presented to track residual stresses incorporating conversion and temperature dependent material property changes. Finally, the relative contribution of thermal- and conversion-dependent strains to residual stresses is evaluated and it is found that the conversion-based strains are the major contributor to the overall behavior. This framework provides the basis for understanding the complex development of residual stresses and can be used as the basis for developing more complex models of cement behavior.

Bone Cements↗

Difference in polymerization color changes of dental resin composites by the measuring aperture size.

The objective of this study was to evaluate the influence of the illuminating/viewing aperture diameter of 11 mm/8 mm and 3 mm/3 mm and measuring geometry on the color and polymerization color changes of dental resin composites. Color of resin composites of 12 mm in diameter was measured on a reflection spectrophotometer with SCE and SCI geometry under D65 illumination. Before polymerization, difference in the three-color coordinates by the material was prominent in spite of the fact that the shade designation of all materials was the same. After polymerization, L* value decreased in hybrid composites but not in packable composites, and changes in a* value were small. When the same aperture size was used, polymerization color change with the SCE was generally higher than those with the SCI. Measured color was different by the aperture size, and the color difference was 16.82-27.05 before polymerization and 20.09-28.51 after polymerization. General trends of polymerization color change were similar in both aperture sizes. Polymerization color changes with 3-mm aperture size were higher than those with 8-mm aperture size in hybrid resin composites in both geometries (p < 0.05); however, the color change was material dependent in the case of packable resin composites.

Color↗