In this study, in order to analyze how cells recognize biomaterials, mRNA was evaluated on various substrates as shown in the attached HeLa S3 cells. The expressed Heat-Shock Protein (HSP) 70A, 70B, 90 and 47 mRNA were isolated and detected using the RT-PCR method. As a result, 70B, 90 and 47 mRNA expressions varied with differences in the hydrophilicity hydrophobicity of the substrates. HSP mRNAs outcome was induced significantly in the HeLa S3 cells that had adhered onto the hydrophilic surfaces. On the other hand, in the cells that had adhered to the hydrophobic surfaces, HSP mRNAs expressions were low. In the non-adhered HeLa S3 cells the tendency found in HSP 70B and HSP 47 mRNA's expressions was the same as that found in adhered cells, while there were no significant differences in the HSP 90 mRNA expression among either of the samples. We have concluded that HSP mRNAs expression is an important marker in the study of cell-polymer interactions.

Biocompatible Materials↗

PubMedDate not supplied

Immobilization of human thrombomodulin on glass beads and its anticoagulant activity.

Human thrombomodulin (TM) was for the first time immobilized on glass beads by the reaction between the carboxyl group of TM and the amino group of glass beads using water-soluble carbodiimide. Immobilized human TM exhibited both anticoagulant activity and inhibition of platelet aggregation of human blood.

Anticoagulants↗

PubMedDate not supplied

Synthesis and anticoagulant activity of sulfated glucoside-bearing polymer.

Poly(glucosyloxyethyl methacrylate) sulfate [poly(GEMA) sulfate], which contains sulfated glucoside residues, was prepared by the reaction with N,N-dimethylformamide (DMF)/sulfur trioxide (SO3) complex. The degree of sulfation was easily controlled by changing the amount of DMF/SO3 complex added and reaction time. The total human blood clotting time in the presence of poly(GEMA) sulfate was prolonged by increasing the dose or the degree of sulfation of the polymer. The anticoagulant activity of poly(GEMA) sulfate was also compared with that of dextran sulfate, poly(styrenesulfonic acid), poly(vinylsulfuric acid), and heparin. The result suggests that sulfated saccharide residues are essential for endowing anticoagulant activity to synthetic polymer.

Anticoagulants↗

PubMedDate not supplied

Capture of HIV-1 gp120 and virions by lectin-immobilized polystyrene nanospheres.

A lectin such as concanavalin A (Con A) was immobilized on the surfaces of poly(methacrylic acid) branches covered with polystyrene nanospheres with a diameter of 360 nm, which were obtained by the free radical copolymerization of styrene with the poly(tert-butylmethacrylate) macromonomer, followed by hydrolysis. Using Con A-immobilized (0.3 microgram/cm2) nanospheres, the interaction of the nanospheres with HIV-1 was determined by the reduction of the gp120 level and the viral infectivity of the HIV-1 suspensions after a 60 min incubation at room temperatures. Con A-immobilized nanospheres achieved a 95 and a 77% reduction of the gp120 level and the infectivity at a concentration of 0.5 mg/mL, respectively, indicating the effective capture of the gp120 and the virions.

Concanavalin A↗

PubMedDate not supplied

Direct observation of fibrinogen-heparinoid complexes formation using surface plasmon resonance.

We analyzed the binding of heparinoid or heparin with fibrinogen by real-time measurement using surface plasmon resonance technology. Poly(glucosyloxyethyl methacrylate) sulfate [poly(GEMA) sulfate] and dextran sulfate were used as heparinoids. The binding ability of each sulfated polymer was estimated by having each polymer-containing buffer interact with the sensor chip surfaces that had immobilized fibrinogen. Dextran sulfate and poly(GEMA) sulfate showed high affinity to the fibrinogen in this experiment, while the heparin did not. All of the dextran sulfates were desorbed from its surface, while about 30% of the poly(GEMA) sulfate remained on the immobilized fibrinogen upon the addition of NaCl to the buffer which was done in order to analyze the desorption of poly(GEMA) sulfate or dextran sulfate from the surface of the fibrinogen. These data show that the type of binding between fibrinogen-poly(GEMA) sulfate was different from that of dextran sulfate, indicating that the interaction between fibrinogen and poly(GEMA) sulfate was caused not only by an electrostatic but also by a hydrophobic force. These results suggest that the interaction mechanism of heparinoids with fibrinogen was different from that of heparin.

Biopolymers↗

PubMedDate not supplied

Experimental study of bioactive polyurethane sponge as an artificial trachea.

An artificial trachea that has tissue reconstructive activity and elasticity that matches a biologic trachea was fabricated using a polyurethane sponge (PS). The inner surfaces of the PS were modified by immobilizing collagen, fibronectin, RGD peptide, a coating of collagen, and apatite. Two PS pore sizes were prepared, 150 to 350 microns (small pore) and 350 to 710 microns (large pore). When they were implanted in the subcutaneous tissue of rats, study of the implantation showed severe inflammatory cell infiltration with the treated PS, especially with collagen and collagen coating. Infiltration and maturity of fibroblasts inside the PS were noted with fibronectin. Infiltration of inflammatory cells and fibroblasts was noted with small pore PS compared with large pore PS. Partial tracheal defects were patched with PS. Better epithelization of PS patches was observed with fibronectin and apatite compared with the others. The artificial trachea made of PS was transplanted into the tracheas of dogs. No separation of the anastomosis and tissue reconstruction of the artificial trachea wall were observed; however, sputum absorption and tracheal granulation were seen.

Amino Acid Sequence↗

PubMedDate not supplied

Immobilization of human thrombomodulin onto biomaterials. Comparison of immobilization methods and evaluation of antithrombogenicity.

Human thrombomodulin (hTM), which is a newly described endothelial cell associated protein that functions as a potent natural anticoagulant by converting thrombin from a procoagulant protease to an anticoagulant, was immobilized on to various substrates by two immobilization methods. As the substrates of immobilization, poly(acrylic acid) surface grafted poly(ethylene) (PAAc-g-PE) film, poly(vinylamine) surface grafted poly(ethylene) film, and PAAc surface grafted nylon were used. For immobilization, simultaneous preactivation methods were used. The effect of the immobilization reaction on hTM activities, the comparison of the activities of immobilized hTM with those of free hTM, and the effect of the thrombin incorporation on antithrombogenic activity were studied. The hTM immobilized onto PAAc-g-PE by preactivation showed the highest antithrombogenic activity. The thrombin incorporation affected protein C activation activity but not the fibrinogen clotting time. hTM immobilized nylon showed greater antithrombogenicity in vitro.

Anticoagulants↗

PubMedDate not supplied

In vivo and ex vivo evaluation of the antithrombogenicity of human thrombomodulin immobilized biomaterials.

Human thrombomodulin (hTM) is a newly described endothelial cell associated protein that functions as a potent natural anticoagulant by converting thrombin from a procoagulant protease to an anticoagulant. Focusing on the establishment of the practical evaluation of hTM immobilized materials, the activity of immobilized hTM was evaluated by in vivo and ex vivo blood contacting tests. As the basis for immobilization, regenerated cellulose films and hollow fibers were used. For the in vivo test, hTM immobilized cellular hollow fibers were implanted into dog blood vessels. Using hTM immobilized cellulose hollow fibers, a small scale dialyzer was assembled and its antithrombogenic activity was studied using human blood. As a result, it was revealed that the immobilized hTM still has co-enzymatic activity for activation of Protein C and anticoagulant activity. The coagulation time of the human blood passed through the hTM immobilized small dialyzer was effectively prolonged. It is expected that hTM immobilized cellulose should be a useful antithrombogenic biomaterial.

Animals↗

PubMedDate not supplied

In vitro production of myeloperoxidase anti-neutrophil cytoplasmic antibody and establishment of Th1-type T cell lines from peripheral blood lymphocytes of patients.

OBJECTIVE: To investigate the pathogenic role of T cells in the development of anti-neutrophil cytoplasmic antibody (ANCA) associated vasculitis. METHODS: Peripheral blood lymphocytes (PBL) were isolated from myeloperoxidase anti-neutrophil cytoplasmic antibody (MPO-ANCA) associated vasculitis patients and cultured in medium. The production of MPO-ANCA in the medium of PBL stimulated with Concanavalin-A (Con-A), with or without cyclosporin (CyA), was measured by enzyme-linked immunosorbent assay (ELISA) on MPO coated plates. RNA isolated from PBMC of one patient was used for polymerase chain reaction (PCR) and single stranded conformational polymorphism (SSCP) studies, and MPO-specific T cell lines (TCL) were established by antigen stimulation techniques. RESULTS: PBL of patients with MPO-ANCA-associated vasculitis produced MPO-ANCA following Con-A stimulation, and this effect was inhibited by treatment with cyclosporin A (CyA) or elimination of CD4 cells. PCR-SSCP showed autoantigen-reactive oligoclonal T-cell accumulation in PBMC of one of these patients. We established MPO-specific TCL which secreted interferon-gamma (IFN-gamma), but not interleukin-4 (IL-4); all TCL were CD4 positive, CD8 negative, and HLA-DR restricted. CONCLUSIONS: Our results suggest that Th1-type T cells may mediate MPO-ANCA production, and may play a role in the onset of MPO-ANCA vasculitis.

Aged↗

PubMedDate not supplied

[Mechanism of the anaphylactic desensitization in guinea pigs].

Explore the source record for details and available documents.

Anaphylaxis↗

PubMedDate not supplied

Mutant p53 proteins behave in a dominant, negative fashion in vivo.

The p53 encodes a cellular phosphoprotein that has been association with both neoplastic transformation and the control of cellular growth. Recent studies have reported that p53 also acts as a transcriptional regulator. We have studied transactivational properties of human wild-type and mutant p53 proteins representing 4 major mutational hotspots (codons 141, 175, 248, 273) as well as a double mutant Tyr141/His273 and a p53 with the transcriptional activating region removed (pcDC2). Transactivation by p53 was shown with a p53 concensus binding sequence controlled CAT reporter gene, and activity was assayed after co-transfection of the reporter with either wild-type or mutant p53 expression constructs. Wild-type p53 as well as one mutant p53 [(mutation of arginine to histidine at codon 273 (His 273)], had strong transactivating activity, but all other mutant p53s were inactive in transcriptional activation, including the double mutant Tyr141/His273 suggesting that the Tyr141 mutation was dominant over the His273 mutation in the same protein. Moreover, when mutant p53 (Tyr141, His175, Trp248, or Tyr141/His273) was cotransfected with either wild-type p53 or mutant His273 p53, these mutants inhibited the transactivation of coexpressed wild-type p53. The p53 vector (pcDC2), which contains p53 oligomerization sequences, but not the transactivational domain, markedly inhibited wild-type p53 transactivational activity. Each of the mutant p53s similarly inhibited the transactivation of His273 p53. Therefore, with the exception of His273, each of the other mutant p53 were unable to transactivate and each behaved in a dominant negative fashion.

Base Sequence↗