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PubMed2003

Adhesive surface proteins of Erysipelothrix rhusiopathiae bind to polystyrene, fibronectin, and type I and IV collagens.

Erysipelothrix rhusiopathiae is a gram-positive bacterium that causes erysipelas in animals and erysipeloid in humans. We found two adhesive surface proteins of E. rhusiopathiae and determined the nucleotide sequences of the genes, which were colocalized and designated rspA and rspB. The two genes were present in all of the serovars of E. rhusiopathiae strains examined. The deduced RspA and RspB proteins contain the C-terminal anchoring motif, LPXTG, which is preceded by repeats of consensus amino acid sequences. The consensus sequences are composed of 78 to 92 amino acids and repeat 16 and 3 times in RspA and RspB, respectively. Adhesive surface proteins of other gram-positive bacteria, including Listeria monocytogenes adhesin-like protein, Streptococcus pyogenes protein F2 and F2-like protein, Streptococcus dysgalactiae FnBB, and Staphylococcus aureus Cna, share the same consensus repeats. Furthermore, the N-terminal regions of RspA and RspB showed characteristics of the collagen-binding domain that was described for Cna. RspA and RspB were expressed in Escherichia coli as histidine-tagged fusion proteins and purified. The recombinant proteins showed a high degree of capacity to bind to polystyrene and inhibited the binding of E. rhusiopathiae onto the abiotic surface in a dose dependent manner. In a solid-phase binding assay, both of the recombinant proteins bound to fibronectin, type I and IV collagens, indicating broad spectrum of their binding ability. It was suggested that both RspA and RspB were exposed on the cell surface of E. rhusiopathiae, as were the bacterial cells agglutinated by the anti-RspA immunoglobulin G (IgG) and anti-RspB IgG. RspA and RspB were present both in surface-antigen extracts and the culture supernatants of E. rhusiopathiae Fujisawa-SmR (serovar 1a) and SE-9 (serovar 2). The recombinant RspA, but not RspB, elicited protection in mice against experimental challenge. These results suggest that RspA and RspB participate in initiation of biofilm formation through their binding abilities to abiotic and biotic surfaces.

Amino Acid Sequence↗

PubMed2002

Isolation of an Escherichia coil strain mutant unable to form biofilm on polystyrene and to adhere to human pneumocyte cells: involvement of tryptophanase.

Escherichia coli adherence to biotic and abiotic surfaces constitutes the first step of infection by promoting colonization and biofilm formation. The aim of this study was to gain a better understanding of the relationship between E. coli adherence to different biotic surfaces and biofilm formation on abiotic surfaces. We isolated mutants defective in A549 pneumocyte cells adherence, fibronectin adherence, and biofilm formation by random transposition mutagenesis and sequential passages over A549 cell monolayers. Among the 97 mutants tested, 80 were decreased in biofilm formation, 8 were decreased in A549 cells adherence, 7 were decreased in their adherence to fibronectin, and 17 had no perturbations in either of the three phenotypes. We observed a correlation between adherence to fibronectin or A549 cells and biofilm formation, indicating that biotic adhesive factors are involved in biofilm formation by E. coli. Molecular analysis of the mutants revealed that a transposon insertion in the tnaA gene encoding for tryptophanase was associated with a decrease in both A549 cells adherence and biofilm formation by E. coli. The complementation of the tnaA mutant with plasmid-located wild-type tnaA restored the tryptophanase activity, epithelial cells adherence, and biofilm formation on polystyrene. The possible mechanism of tryptophanase involvement in E. coli adherence and biofilm formation is discussed.

Bacterial Adhesion↗

PubMed2006

Spontaneous condensation in DNA-polystyrene- b-poly(l-lysine) polyelectrolyte block copolymer mixtures.

We investigated the condensation of calf thymus DNA by amphiphilic polystyrene(m)-b-poly(l-lysine)(n) block copolymers ( PS(m)-b- PLys(n), m, n = degree of polymerization), using small-angle X-ray scattering, polarized optical microscopy and laser scanning confocal microscopy. Microscopy studies showed that the DNA condenses in the form of fibrillar precipitates, with an irregular structure, due to electrostatic interactions between PLys and DNA. This is not modified by the presence of hydrophobic PS block. Scattering experiments show that the structure of the polyplexes corresponds to a local order of DNA rods which becomes more compact upon increasing n. It can be concluded that for DNA/ PS(m)-b- PLys(n) polyplexes, the balance between the PLys block length and the excess charge in the system plays an essential role in the formation of a liquid crystalline phase.

Binding Sites↗

PubMed2003

Inhibition of prostaglandin synthesis during polystyrene microsphere-induced pulmonary embolism in the rat.

Our objective was to test the effect of inhibition of thromboxane synthase versus inhibition of cyclooxygenase (COX)-1/2 on pulmonary gas exchange and heart function during simulated pulmonary embolism (PE) in the rat. PE was induced in rats via intrajugular injection of polystyrene microspheres (25 micro m). Rats were randomized to one of three posttreatments: 1) placebo (saline), 2) thromboxane synthase inhibition (furegrelate sodium), or 3) COX-1/2 inhibition (ketorolac tromethamine). Control rats received no PE. Compared with controls, placebo rats had increased thromboxane B(2) (TxB(2)) in bronchoalveolar lavage fluid and increased urinary dinor TxB(2). Furegrelate and ketorolac treatments reduced TxB(2) and dinor TxB(2) to control levels or lower. Both treatments significantly decreased the alveolar dead space fraction, but neither treatment altered arterial oxygenation compared with placebo. Ketorolac increased in vivo mean arterial pressure and ex vivo left ventricular pressure (LVP) and right ventricular pressure (RVP). Furegrelate improved RVP but not LVP. Experimental PE increased lung and systemic production of TxB(2). Inhibition at the COX-1/2 enzyme was equally as effective as inhibition of thromboxane synthase at reducing alveolar dead space and improving heart function after PE.

Angiography↗

PubMed1997

Kayexalate (sodium polystyrene sulphonate) in sorbitol associated with intestinal necrosis in uremic patients.

BACKGROUND: Kayexalate (sodium polystyrene sulphonate) in sorbitol is commonly used to treat hyperkalemia in patients with renal insufficiency. Isolated case reports and one recent large series have documented intestinal necrosis following administration of kayexalate in sorbitol. METHODS: Two patients with luminal kayexalate crystals associated with intestinal pathology were first identified in the pathology department, and clinicopathological correlation was carried out. RESULTS: Both patients were seriously ill, had prior cardiac surgery and were in renal failure (uremic). Examination of autopsy and colonic resection showed luminal kayexalate crystals associated with underlying mucosal necrosis, submucosal edema and transmural inflammation. CONCLUSION: Although occurring in complex clinical settings, the pathological findings provide additional evidence that kayexalate in sorbitol may be associated with intestinal necrosis and inflammation in uremic patients and that this may be a clinically and pathologically under-recognized iatrogenic bowel injury.

Aged↗

PubMed1985

Antibody determination against Aspergillus fumigatus by means of the enzyme-linked immunosorbent assay. I. Liberation of polystyrene binding components during cultivation.

The liberation of polystyrene binding antigenic components (PBC) of Aspergillus fumigatus involved in ELISA IgG measurements was studied during cultivation in the culture filtrates. PBC were rapidly liberated into the culture medium during the logarithmic phase of growth (phase I), were in part degraded or reconsumed during the stationary phase (phase II) and again secreted (other PBC) during the lytic phase (phase III). ELISA IgG titers and numbers of precipitating components are not always related with one another, may show an inverse relationship; maximal ELISA titers are reached during phase I when precipitating components are limited. The striking similarity between the liberation of PBC and IgE-binding components is discussed.

Antibodies, Fungal↗

PubMed2004

Anti-prothrombin antibodies as a potential risk factor of recurrent venous thromboembolism.

The role of antiprothrombin (aPT) antibodies in the development of venous thromboembolism (VTE) is still uncertain. The aim of this study was to evaluate the potential role of aPT antibodies in the development of recurrent thromboembolism. Out of 236 consecutive symptomatic patients with an episode of acute VTE, antiphospholipid antibodies were found in 85 (36.0%), of whom 24 were carriers of aPT antibodies (10.2% of the entire cohort). A history of previous thromboembolism was identified in 56 patients (23.7%). The prevalence of previous thromboembolism was significantly higher in carriers than in non-carriers of antiphospholipid antibodies (OR=2.4; 95% CI, 1.3 to 4.4). Of the 24 patients with aPT antibodies, 12 had a history of previous thromboembolism. In a multivariate logistic regression analysis, in which the other categories of antiphospholipid antibodies were taken into account, as well as the patient's age, sex, and the modality of clinical presentation, it was found that the presence of aPT antibodies was significantly associated with the prevalence of prior thromboembolism (OR=3.3; 95% CI, 1.3 to 8.6). Since aPT antibodies are more commonly identifiable in patients with multiple thrombotic episodes, they are a likely risk factor for recurrent thromboembolism.

Adult↗

PubMed2000

Covalent grafting of polythymidylic acid onto amine-containing polystyrene latex particles.

The covalent grafting of a 35-base poly(thymidylic) acid (dT35) onto aminated polystyrene latex has been studied. The oligonucleotide was activated by 1,4-phenylene diisothiocyanate (PDC) prior to grafting reaction. The influence of various experimental conditions related either to the medium (pH, ionic strength) or to the latex (nature and surface charge density, presence of adsorbed nonionic surfactant) was studied. Discrimination between covalent grafting and adsorption was achieved by desorbing the non-covalently grafted dT35 induced by consecutive washings with a suitable buffer solution. The amount of covalently grafted oligonucleotide directly correlated to the concentration and accessible functional groups at the particle surface.

Adsorption↗

PubMed2000

Surface modification of polystyrene nanoparticles using dextrans and dextran-POE copolymers: polymer adsorption and colloidal characterization.

Hydrophobically-modified dextran (dextran-phenoxy, DexP) and dextran-phenoxy-poly(oxyethylene) (DexP-POE) copolymers have been used to modify the surface properties and the stability of polystyrene nanoparticles. We examined the effect of phenoxy group and POE chain concentrations on their adsorption behaviour. The adsorbed amount was determined by the standard depletion method and the layer thickness of the adsorbed layer by photon correlation spectroscopy and electrokinetic measurements. The results show that the hydrophobic interaction is the driving force during the adsorption while the layer thickness correlates with the interfacial concentration of grafted POE chains. The effects of adsorbed layers on the properties of latex dispersions have been characterized in terms of the stability of the dispersions toward added electrolyte and temperature. The conformation of the adsorbed copolymers is discussed in relation to layer thickness and colloidal stability of suspensions.

Adsorption↗

PubMed2000

The use of immunoliposomes for specific delivery of antimicrobial agents to oral bacteria immobilized on polystyrene.

Antibacterial immunoliposomes have been prepared using covalently bound antibody, raised to the cell surface of the bacterium Streptococcus oralis (S. oralis), and incorporating the bactericides chlorhexidine and Triclosan. A regrowth assay, in which the ability of a bacterial biofilm immobilised on polystyrene to grow after exposure to a test solution, was undertaken to study the action of the antibacterial immunoliposomes. The antibacterial anti-oralis immunoliposomes show enhanced growth inhibition of S. oralis, compared to free bactericide, using low bactericide concentrations. For short exposure times to the biofilms, antibacterial anti-oralis immunoliposomes can show several times enhanced growth inhibition of S. oralis compared to free bactericide. Antibacterial anti-oralis immunoliposomes inhibit the growth of S. oralis more than that of other oral bacteria. The extent of growth inhibition by antibacterial anti-oralis immunoliposomes is linearly related to the number of immunoliposomes targeted to the biofilm surface.

Anti-Infective Agents, Local↗

PubMed2001

Interaction of poly(2-acrylamido 2-methylpropane sulfonate)-grafted polystyrene beads with cationic complement proteins.

Influence of various biomaterials on the complement system in serum has been intensively studied by many research groups, since activation of the complement pathway in vivo has been known to give rise to some pathological conditions, such as inflammation and anaphylaxis. Much effort has been devoted to develop new materials that do not activate or deteriorate the complement system. The present work is aimed at revealing the mode of reactions of anionic poly(2-acrylamido 2-methylpropane sulfonate) grafted on polystyrene bead (PAMPS-g-bead) with serum complement. Complement activity assay, determination of complement proteins levels, and immunoblot analysis were carried out for sera pretreated with PAMPS-g-beads. The results clearly showed that, when PAMPS-g-beads were incubated with serum, those beads adsorbed several complement proteins, i.e. C1q, factor D, factor P, C6, and C8, but the generation of activation fragments of complement components was not observed. Especially, factor D was most effectively removed from serum, resulting in potential inhibition of the alternative pathway. A larger amount of PAMPS-g-beads was needed to decrease the serum CH50 level. That may be caused by removal of C6. Although some polyanions, such as dextran sulfate, were reported to activate the complement system, the obtained results indicate that the PAMPS-g-bead is not an activator of the complement pathway, but acts as an adsorbent of complement components. One possible clinical application of the PAMPS-g-beads is adsorption of serum factor D by extracorporeal treatment of patients with renal failure with a high level of factor D, because the increased quantity of factor D in serum may cause consistent activation of the alternative pathway.

Biocompatible Materials↗

PubMed2005

Passive adsorption of human antirrabic immunoglobulin onto a polystyrene surface.

The latex agglutination immunoassay technique uses polymer colloids as carriers of adsorbed proteins to enhance the antigen-antibody reaction. The aim of the present work is to study the adsorption of Human Antirrabic Immunoglobulin (HA-IgG) on polystyrene latex (PS). The physical adsorption of HA-IgG on PS latex was investigated as a function of pH at 2 mM ionic strength. The amount of HA-IgG adsorbed onto PS latex greatly depends on pH; its value showed a maximum in the neighborhood of the IEP of HA-IgG. The electrophoretic method was applied to characterize latex particles. The influence of the amount of HA-IgG adsorbed (J(ads)) on the electrophoretic mobility and zeta-potential values was also studied.

Adsorption↗

PubMed1991

Interactions of functionalized polystyrene derivatives with the complement system in human serum.

The interactions between blood and insoluble polysaccharidic surfaces result in activation of the immune system of complement. When substituted with carboxymethyl groups, Sephadex loses its capacity to activate complement, whereas Sephadex sulphate has been described as an activator. In order to elucidate the molecular mechanisms of complement activation and inhibition, a simpler polymer model has been chosen: it consists of an insoluble polystyrene backbone on which either isolated hydroxymethyl or sulphonate groups or both are present. The surfaces bearing the isolated groups consume complement but the mechanisms involved are quite different. In contrast, a surface bearing equal proportions of both types of groups is a non-activator. Such model surfaces can be very useful for designing artificial surfaces able to control in situ complement activation.

Biocompatible Materials↗

PubMed1991

The direct determination of protein concentration for proteins immobilized on polystyrene microspheres.

In this paper we show how the Lowry method, designed for the determination of proteins in solution, can be used for the determination of proteins immobilized on the surface of microspheres (for protein concentrations higher than 10 micrograms/ml). Measurements were made for human serum albumin (HSA) and for immunoglobulins [rabbit immunoglobulins--antibodies against human fibrinogen (IgGF) and against fragment D of human fibrinogen (IgGFgD)] immobilized on the surface of polystyrene microspheres.

Animals↗

PubMed1993

Mechanism of cytoplasmic calcium changes in platelets in contact with polystyrene and poly(acrylamide-co-methacrylic acid) surfaces.

Changes in cytoplasmic free calcium levels ([Ca2+]i) in platelets in contact with polystyrene (PSt) and poly(acrylamide-co-methacrylic acid) (PAAmMAc) particles were evaluated and results were compared with those from two representative biological calcium agonists; thrombin and calcium ionophore A23187. PSt particles stimulated a steep increase in cytoplasmic calcium levels in platelets as much as thrombin and A23187. Serratia protease-treated platelets showed a steep increase in [Ca2+]i by PSt particles, suggesting that PSt surfaces can initiate platelet activation independent of a glycoprotein Ib (GPIb)-mediated pathway. By contrast, dibucaine-treated platelets showed little increase in [Ca2+]i by PSt particles, indicating that microfilament assembly, including binding of GPIb with actin binding protein, should be required for platelet activation in contact with PSt surfaces. PAAmMAc particles induced little increase in cytoplasmic calcium levels in platelets. However, PAAmMAc particle-treated platelets demonstrated little response to thrombin in terms of an increase in [Ca2+]i and ATP release, suggesting the possibility that PAAmMAc surfaces may regulate [Ca2+]i by influencing platelet metabolism. Furthermore, sodium azide-treated platelets showed an increase in [Ca2+]i in platelets when contacting PAAmMAc particles, supporting the suggestion that PAAmMAc surfaces could regulate platelet functions. Fluorescence polarization measurements using 1,6-diphenyl-1,3,5-hexatriene-loaded platelets revealed that PAAmMAc particles increased membrane fluidity in platelets, which may be due to physicochemical interaction with PAAmMAc surfaces.

Acrylic Resins↗

PubMed1994

Monosize microbeads based on polystyrene and their modified forms for some selected medical and biological applications.

Polymeric particles are produced by different polymerization techniques. Phase inversion (dispersion) polymerization is one of the recent techniques to obtain monosize polymeric microbeads in the size range of 1-50 microns. The size and monodispersity of these microbeads can be adjusted by using several solvent systems (e.g., alcohol-water mixtures) with different polarities and by changing the type and amount of monomer, initiator and stabilizer. Surfaces of these microbeads can be further modified by different techniques including coating with different copolymers. Monosize polymeric microbeads are widely used in medical and biological applications as carriers, such as in immunoassays and cell separation, in site-specific drug delivery systems, in nuclear medicine for diagnostic imaging, in studying the phagocytic process, in affinity separation of biological entities, etc. Here, some important aspects of the production of monosize microbeads based on polystyrene and their modified forms are briefly discussed, and some selected medical and biological applications are summarized.

Biopolymers↗

PubMed1994

Antithrombogenicity of hydrophilic polyurethane-hydrophobic polystyrene IPNs. I. Synthesis and characterization.

A series of interpenetrating polymer networks (IPNs) composed of hydrophilic polyurethane (PU) and hydrophobic polystyrene (PS) were prepared by the simultaneous polymerization method. The PU network was synthesized via the isocyanate-terminated PU prepolymer based on polyethylene glycol (PEG), a highly hydrophilic oligomer, and hexamethylene diisocyanate (HDI). The bulk and surface characteristics of these materials were analyzed by differential scanning calorimetry (DSC), tensile testing, scanning electron microscopy (SEM), attenuated total reflectance-Fourier transform infrared (ATR-FTIR), electron spectroscopy for chemical analysis (ESCA), and contact angle measurement. The PU/PS IPNs prepared in this study exhibited phase separated structures, which had dispersed PS domains in the continuous PU matrix, in both the bulk and surface showing two transition temperatures. The IPN containing 50 wt% of PS showed good mechanical properties. The enrichment of PU phase in the surface was revealed by SEM, ATR-FTIR, ESCA, and contact angle measurement.

Biocompatible Materials↗

PubMed1994

Antithrombogenicity of hydrophilic polyurethane-hydrophobic polystyrene IPNs. II. In vitro and ex vivo studies.

To investigate the effect of hydrophilic and hydrophobic surfaces with phase separated structure on their blood responses, interpenetrating polymer networks (IPNs) composed of hydrophilic polyurethane (PU) and hydrophobic polystyrene (PS) were prepared by simultaneous polymerization. In vitro protein adsorption, in vitro platelet adhesion, and ex vivo A-A shunt test were carried out to evaluate the blood compatibility of the PU/PS IPNs. The results of protein adsorption on the PU/PS IPN surfaces indicated that albumin preferentially adsorbed on the hydrophilic surface (PU), while fibrinogen preferentially adsorbed on the hydrophobic surface (PS). The PU/PS IPNs exhibited suppressive properties for both platelet adhesion and activation. The occlusion time of U50S50 IPN containing 50 wt% of PS was twice as long as that of the PU control (50 min), indicating enhanced blood compatibility, presumably due to the selective adsorption of plasma proteins and the suppression of the adhesion and activation of platelets.

Adsorption↗