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PubMed2006

Application of polymeric surfactants in micellar electrokinetic chromatography-electrospray ionization mass spectrometry of benzodiazepines and benzoxazocine chiral drugs.

Chiral micellar EKC (CMEKC) coupled to ESI-MS using polymeric surfactants as pseudostationary phases is investigated for simultaneous enantioseparation of two benzodiazepines, (+/-)-oxazepam ((+/-)-OXA) and (+/-)-lorazepam ((+/-)-LOR), and one benzoxazocine, (+/-)-nefopam ((+/-)-NEF). First, enantioselectivity and electrospray sensitivity of six chiral polymeric surfactants for all three chiral compounds are compared. Second, using poly(sodium N-undecenoyl-L-leucinate) as pseudostationary phase, the organic modifiers (methanol (MeOH), isopropanol, and ACN) are added into the running buffer to further improve chiral resolution (RS). Next, a CMEKC-ESI-MS method for the simultaneous enantioseparation of two benzodiazepines is further developed by using a dipeptide polymeric surfactant, poly(sodium N-undecenoxy carbonyl-L,L-leucyl-valinate) (poly-L,L-SUCLV). The CMEKC conditions including nebulizer pressure, capillary length, ammonium acetate concentration, pH, poly-L,L-SUCLV concentration, and capillary temperature were optimized to achieve maximum chiral RS and highest sensitivity of MS detection. The spray chamber parameters (drying gas temperature and drying gas flow rate) as well as sheath liquid conditions (MeOH content, pH, flow rate, and ionic strength) were found to significantly influence MS S/N of both (+/-)-OXA and (+/-)-LOR. Finally, a comparative study between simultaneous UV and MS detection showed high plate numbers, better chiral RS, and enhanced detectability with CMEKC-MS. However, speed of analysis was faster using CMEKC-UV.

Benzodiazepines↗
PubMed2006

Permanent surface modification of polymeric capillary electrophoresis microchips for protein and peptide analysis.

Because of their surface heterogeneity, proteins readily adsorb on polymeric substrates via various interactions, which adversely affects the performance of polymeric microfluidic devices in electrophoresis-based protein/peptide analysis. Therefore, it is necessary to use surface modification techniques such as dynamic coating or more complicated permanent surface modification, which has broader application and better performance, to render the polymeric microchannels protein-resistant. This manuscript is a review of the surface chemistry of microfluidic devices used for electrophoretic separations of proteins and peptides. The structural complexity of proteins as it relates to adsorption is described, followed by a review of the mechanisms and structural characteristics of protein-resistant surfaces. Permanent surface modification techniques used in grafting protein-resistant materials onto the surfaces of electrophoresis microchannels fabricated from polymer substrates are summarized and successful examples are presented.

Electrophoresis, Capillary↗
PubMed1984

Localization of two phosphorylation sites adjacent to a region important for polymerization on the tail of Dictyostelium myosin.

Phosphorylation of the myosin heavy chains of Dictyostelium discoideum is known to be inhibited following chemotactic stimulation of the cells. Effects of dephosphorylation on the assembly of myosin and on its actin-activated ATPase activity raised the question of where the phosphorylated sites are located with respect to sites responsible for polymerization and actin binding. Using seven monoclonal antibodies the binding sites of which were mapped in the electron microscope, two phosphorylation sites, i.e., threonine residues that were phosphorylated by a kinase from D. discoideum, were localized by immunoblotting of chymotryptic fragments. Two of the antibodies bound to the terminal one fifth of the tail and recognized a phosphorylated chymotryptic fragment of 38 kd. The non-phosphorylated form and single and double phosphorylated forms of this fragment were separated by two-dimensional electrophoresis. Antibody labeling of lower mol. wt. polypeptides indicated that both phosphorylation sites were located at least 32 kd from the end of the tail. A non-phosphorylated fragment, that was insoluble at low ionic strength due to polymerization, proved to be an internal cleavage product of the tail. A segment of this fragment necessary for polymerization was mapped adjacent to the phosphorylation sites.

Journal Article↗
PubMed2002

The influence of system compliance and sample geometry on composite polymerization shrinkage stress.

The objective of this study was to use finite-element analysis to model tensilometer tests of polymerizing dental composites. A typical sample in polymerization shrinkage stress tests is shaped like a flat disk, that is, has a high aspect ratio (ratio of diameter to height). In the experimental literature it is implied that the induced stress state in the flat disk composite samples is uniaxial. Three published tensilometer tests of curing dental composite samples with similar high aspect ratios (varying from 3 to 5) were modeled, but with test configurations having low, intermediate and high relative compliance (a tenfold variation). With the use of linear elastic finite element analysis, an instantaneous volumetric shrinkage of 1% was applied to the composite via the thermal analogy and the following questions were addressed: 1. Does the numerically predicted state of stress in composite samples tested in this fashion differ substantially from the uniaxial stress state assumed in the experiments?2. How do the numerically predicted stresses compare with the experimentally determined nominal stresses?3. Does compliance of the mountings influence the numerically predicted stress state? The finite-element results predicted a complex triaxial stress state that was strongly influenced by the compliance of the mountings. For the low and intermediate system compliance the model overpredicted the polymerization contraction stress, as would be anticipated due to the inability of the model to account for viscoplastic flow. For high system compliance, the numerical and experimental stress values were in better agreement, mainly because the linear elastic mountings accounted for most of the measured system compliance.

Compliance↗
PubMed1979

Bioevaluation of plasma polymerized films in skeletal muscle.

Plasma polymerized ethylene (PPE), styrene (PPS), and chlorotrifluoroethylene (PPCTFE) were synthesized by exposing the monomeric gases to an inductively coupled radio frequency "glow-discharge" field. The polymer films were deposited on poly(dimethyl) siloxane (medical grade Silastic), which was then surgically implanted in rat paravertebral muscle for periods up to 84 weeks. The biocompatibility of the plasma deposited films and uncoated Silastic was evaluated by qualitative (graded inflammatory cell response) and quantitative (connnective tissue capsule thickness) techniques as a function of time. The morphological features of the connective tissue capsule and the plasma polymerized films were examined by SEM after 75 weeks of implantation. Results showed that the acute inflammatory cell migration around PPS and PPCTFE was at a maximum in 2 weeks, decaying to control levels in 4 to 8 weeks. The PPE response was judged as less than the control response up to 4 weeks. After 8 weeks no qualitative difference could be detected between the plasma polymerized films and Silastic. On the other hand, a quantifiable change in fibrous capsule response as a function of time and material was noted until 24 weeks. From these data we conclude that these types of films do not elicit an untoward foreign body reaction at a skeletal muscle implant site in rats.

Animals↗
PubMed1983

Polymerization kinetics of resin-based restorative materials.

The kinetics and mechanism of cure of resin-based restorative materials have been investigated by incorporating into commercial materials additional amounts of inhibitor, initiator, and accelerator. The polymerization was monitored by IR spectroscopy, viscosity measurements, and an oscillating rheometer. The rate of initiation of the polymerization was found to be first order with respect to the initiator and accelerator concentration. The inhibitor was found to be responsible for the induction period during which no polymerization occurred. The duration of this period was proportional to the inhibitor concentration. It was also found that the efficiency of the inhibitor affected the difference between the initial and final set times so that with one particularly effective inhibitor a "snap-set" behavior could be obtained.

Benzoquinones↗
PubMedDate not supplied

Characterization of plasma polymerized silicone coatings useful as biomaterials.

Plasma polymerization techniques were used to deposit a layer of filler-free silicone rubber on a variety of substrate materials. The thickness of the deposited film was 0.5-0.8 micron. As it is the surface of the biomaterial that comes in direct contact with the body fluids, the surface of the biomaterial is of paramount importance. In this study, the plasma polymerized biomaterials were characterized. Thus, the scanning electron microscope (SEM) showed the surfaces to be smooth. To study the surface layer of the deposited polymer, Fourier-transform infrared spectrometry in the attenuated total reflection (ATR) mode was used. The deposited material was indeed silicone polymer with adsorption bands at 1262, 1020, and 802 cm-1 for the Si-CH3 bending, Si-0-Si stretching, and Si-CH3 bending, respectively. To find the bonding nature of the polymer, electron spectrometry for chemical analysis (ESCA) was used. The silicone polymer was shown to be highly cross-linked. To find the molecular weight between cross-links, swelling studies were done. Thus the results of the study show that the plasma polymerization could produce a filler-free silicone layer on a variety of substrate materials.

Biocompatible Materials↗
PubMed1988

Blood compatibility of surfaces modified by plasma polymerization.

Tubular blood-contacting polymeric materials were modified by plasma polymerization and evaluated in the baboon with respect to their capacity to induce both acute and chronic arterial thrombosis. Polymer surface composition was determined by electron spectroscopy for chemical analysis. Steady-state arterial thromboembolism was initiated by introducing tubular segments into chronic arteriovenous shunts. Rates of platelet destruction induced by the test materials were calculated from 111In-platelet survival measurements. Nine plasma polymers based on tetrafluoroethylene, hexafluoroethane, hexafluoroethane/H2, and methane, when deposited on silicone rubber, consumed platelets at rates ranging from 1.1-5.6 x 10(8) platelets/cm2-day. Since these values were near the lower detection limit for this test system, the plasma polymers were considered relatively nonthrombogenic. Acute thrombus formation was initiated by inserting expanded Teflon (Gore-Tex PTFE) vascular grafts into the shunt system. 111In-platelet deposition was measured by scintillation camera imaging over a 1-h exposure period. Standard PTFE grafts (10 cm x 4 mm i.d.) accumulated approximately 1 x 10(10) platelets over this interval. While modification of PTFE grafts with a plasma polymer based on hexafluoroethane/H2 did not alter graft surface morphology, platelet deposition was reduced by 87% as compared to the controls (p less than 0.001). We conclude that both the surface chemistry and texture of prosthetic materials influence thrombogenesis. The method of plasma polymerization may be useful for assessing the importance of these variables independently and, perhaps, for minimizing certain adverse blood-material interactions.

Animals↗
PubMed1990

A hydrophilic plasma polymerized film composite with potential application as an interface for biomaterials.

A hydrophilic polymer composite film (approx. 420 nm thick), with potential application as an interface for biomaterials has been prepared on nonorganic substrates, which include glass, silicon, and aluminum foil, using a glow discharge plasma polymerization technique. A thin film (110 nm thick) polymerized from hexane provided an adherent protective coating for the substrate material, and covalent bonding sites for the outer layer polymerized from N-vinyl-2-pyrrolidone. This outer layer provided the hydrophilic surface or interface. The two layers were copolymerized for a short period during transition between monomers to provide an intimate covalently bonded diffuse interphase. Preliminary in vitro and in vivo biocompatibility studies indicate that the hydrophilic film is non-cytotoxic, and does not increase the inflammatory response when compared with negative controls.

Animals↗
PubMed1995

Biocompatibility study of as-polymerized poly(L-lactide) in rats using a cage implant system.

To evaluate the biocompatibility of in vitro predegraded as polymerized poly(L-lactide) (PLLA), a cage implant system was used to investigate white cell and enzyme concentrations with time. The use of a cage permits in a serial fashion a quantitative and qualitative measurement of exudate components formed around an implant. Subcutaneously in rats, caped cages manufactured from stainless-steel mesh were implanted with in vitro predegraded, as-polymerized PLLA, as-polymerized PLLA cylinders, and empty cages serving as controls. In vitro predegradation was used to simulate the degradation products of long-term in vitro degradation. Predegraded PLLA particles were obtained by in vitro hydrolysis at elevated temperatures. The first 7 days of implantation were characterized by an acute inflammatory reaction; the exudate extracted from the cages showed predominantly neutrophils for all types of implants. After day 7, there was a more chronic inflammatory reaction with predominantly macrophages and lymphocytes. There were no significant differences in the total leukocyte concentration or macrophage concentration for any of the cages in the period from 10-21 days. Extracellular enzyme activity also did not show any significant differences among the three types of cages. A possible explanation for the absence of any significant differences could be that the in vitro predegraded particles were sieved before implantation, thus eliminating all small particles (< 70 microns) that are probably mandatory to provoke an increased cellular reaction.

Animals↗
PubMed2005

Preparation of thermosensitive microgels via suspension polymerization using different temperature protocols.

A thermosensitive and biodegradable microgel for protein drug release was synthesized from a thermosensitive macromer via inverse suspension polymerization. Preparation was made under a constant temperature or under variable temperatures. In the latter protocol, dispersion was performed at a low temperature below lower critical solution temperature of the macromer aqueous solution and polymerization was performed at a high temperature above lower critical solution temperature. According to the experiments, the constant-temperature method was not suitable for preparation of microgels when the macromer concentration was high, because early physical gelation at the preparation temperature seriously influenced formation of dispersed droplets. If the macromer concentration was low, both temperature protocols resulted in spherical hydrogel microparticles, but the properties of the resulting microgels were different to a certain degree. In both cases, the model protein bovine serum albumin was loaded into microgels by a postfabrication encapsulation technique, which takes advantage of the microgels' negative thermosensitivity. The results demonstrate that, in microgel preparation, the variable-temperature protocol is useful in suspension polymerization of negatively thermosensitive macromers at a wide rage of monomer concentrations.

Animals↗
PubMed2007

A novel synthetic tissue-adhesive hydrogel using a crosslinkable polymeric micelle.

We prepared a novel tissue-adhesive hydrogel by using a polymeric micelle consisting of an aldehyde-terminated poly(ethylene glycol)-poly(D,L-lactide) (PEG-PLA) block polymer. A Schiff base is chemically formed between the amino groups in a polyallylamine and the aldehyde groups on the surface of polymeric micelles. The hydrogel was formed in approximately 2 s when the polymeric micelle solution and polyallylamine solution are mixed in vitro. The hydrogel was rapidly formed in vivo, and it adhered to a tissue surface. Our novel tissue-adhesive hydrogel creates no risk of infectious contaminations, because it consists of only synthetic materials. Further, PEG and PLA are known to be biocompatible and noncytotoxic. The results obtained in the present study show that a hydrogel prepared by the formation of a Schiff base between aldehyde and amine groups will potentially address the need for novel tissue-adhesive materials.

Cross-Linking Reagents↗
PubMed2006

Evaluation of polymerization of light-curing hybrid composite resins.

The quality of polymerization of hybrid composite resins was tested to explore their feasibility for dental restorations. For this, microhardness, polymerization shrinkage, the coefficient of thermal expansion, and surface morphology were evaluated during or after light curing in conjunction with the thermocycling process. Each product had different microhardness values. The repeated thermal stimulus has no specific effect on the change of microhardness. The difference of microhardness between the thermocycled specimens and specimens stored only in distilled water was minor. The measured microhardness had a linear correlation with the filler content (vol %) of the tested specimens. The polymerization shrinkage had rapidly increased only during the light curing, and then it reached a plateau. Among the specimens, Z250 showed the least amount of shrinkage for all tested thicknesses. Regardless of the product, the shrinkage values increased as the specimens became thick. The coefficient of thermal expansion of the control specimens ranged between 42 and 55 microm/degrees C in the temperature range of 30-80 degrees C. The coefficient showed an inverse correlation with the filler content. Through the thermocycling process, Palfique Estelite showed randomly propagating cracks on the surface. Larger fillers showed a more apparent detachment than the smaller fillers.

Hot Temperature↗
PubMed2007

Influence of changes in the composition of an acrylic bone cement on its polymerization kinetics.

It has been suggested in the literature that a lower polymerization rate of an acrylic bone cement is favorable for the in vivo longevity of a cemented arthroplasty. The present work was a study of the influence of three changes in the composition of an acrylic bone cement (when taken separately) on the cement polymerization rate at 37 degrees C (assumed to be the temperature in the bone bed during a cemented arthroplasty) [k']. The changes were the amount of copolymer as a proportion of the total powder weight (in cements in which there is a copolymer in the powder), the amount of DMPT as a proportion of the total volume of the liquid monomer, and the accelerator. k' was calculated using values of the activation energy and the frequency factor (assuming the polymerization reaction is Arrhenius in nature) that were computed from measurements made using the nonisothermal mode of differential scanning calorimetry. Statistical analysis (one-way ANOVA, with Bonferroni correction, and factorial ANOVA) of the k' values showed that the change in accelerator had a significant influence on k'. The importance of this finding, together with results from two relevant literature reports, is discussed within the context of the use of modified bone cements in cemented arthroplasties.

Arthroplasty, Replacement↗
PubMed1986

Fibronectin potentiates actin polymerization in thrombin-activated platelets.

The effect of fibronectin on the polymerization state of actin was studied. Triton X-100-insoluble cytoskeleton was prepared from thrombin-activated platelets, and the conversion of G-actin into F-actin was monitored by an assay involving DNase I inhibition by G-actin. It was found that fibronectin bound to membrane receptors decreased the level of platelet G-actin. This observation suggests that in the presence of fibronectin a larger amount of F-actin becomes incorporated into the Triton X-100-insoluble cytoskeleton. At the same molar concentration, fibrinogen only slightly increased actin polymerization, whereas bovine serum albumin at a much higher concentration caused a small inhibition of actin immobilization. Our data show that fibronectin, through interaction with the platelet actomyosin fibrillar system, facilitates actin polymerization into the cytoskeleton.

Actins↗
PubMed1988

Relationship of pseudopod extension to chemotactic hormone-induced actin polymerization in amoeboid cells.

Aggregation-competent amoeboid cells of Dictyostelium discoideum are chemotactic toward cAMP. Video microscopy and scanning electron microscopy were used to quantitate changes in cell morphology and locomotion during uniform upshifts in the concentration of cAMP. These studies demonstrate that morphological and motile responses to cAMP are sufficiently synchronous within a cell population to allow relevant biochemical analyses to be performed on large numbers of cells. Changes in cell behavior were correlated with F-actin content by using an NBD-phallacidin binding assay. These studies demonstrate that actin polymerization occurs in two stages in response to stimulation of cells with extracellular cAMP and involves the addition of monomers to the cytochalasin D-sensitive (barbed) ends of actin filaments. The second stage of actin assembly, which peaks at 60 sec following an upshift in cAMP concentration, is temporally correlated with the growth of new pseudopods. The F-actin assembled by 60 sec is localized in these new pseudopods. These results indicate that actin polymerization may constitute one of the driving forces for pseudopod extension in amoeboid cells and that nucleation sites regulating polymerization are under the control of chemotaxis receptors.

Actins↗
PubMed2002

Integral algorithm and density matrix integration scheme for ab initio band structure calculations on polymeric systems.

A new program for band structure calculations of periodic one-dimensional systems has been constructed. It is distinguishable from other codes by the efficient two-electron integral evaluation and the integration schemes of the density matrix in the first Brillouin zone. The computation of polymeric two-electron integrals is based on the McMurchie Davidson algorithm and builds batches of the different cell indices included in the polymeric system. Consequently it presents efficient scaling with respect to the number of unit cells taken into account. Our algorithm takes into account fully the polymeric symmetry rather than the molecular symmetry. A semidirect procedure where only exchange integrals are computed at each SCF cycle is proposed in order to maintain balance between computation time and disk space. In addition, the integration of the density matrix over a large number of cell indices can be performed by different methods, such as Gauss-Legendre, Clenshaw-Curtis, Filon, and Alaylioglu-Evans-Hyslop. This last scheme is able to obtain an accuracy of 10(-13) a.u. on each individual density matrix element for all cell indices with only 48 k-points.

Journal Article↗
PubMedDate not supplied

Crowding and the polymerization of sickle hemoglobin.

Under physiological conditions, sickle hemoglobin, a natural mutant of human hemoglobin A with a surface hydrophobic valine in place of a negatively charged glutamic acid, polymerizes at high volume occupancy. Equilibrium solubility of sickle hemoglobin entails activity coefficients that can approach 10(3) at high concentrations. Polymerization occurs by homogeneous and heterogeneous nucleation mechanisms, which are both profoundly sensitive to crowding; homogeneous nucleation rates for example are enhanced by 10(10) when the initial concentration is augmented by 50% non-polymerizing hemoglobin. A molecular description of the reaction therefore entails substantial corrections for molecular crowding which are all very accurately described by excluded volume corrections, treating hemoglobin as a hard sphere with volume consistent with the molecular structure of the molecule, and involving no further adjustable parameters. These effects and the descriptions that rationalize this behavior are described.

Anemia, Sickle Cell↗