Heat treatment effects on electrochemical corrosion parameters of high-Pd alloys.

This research determined the effect oxidation, as that occurs during porcelain firing, has upon the corrosion parameters of Pd-based ceramic alloys and how it may relate to Pd allergy. The 20 h open circuit potential (OCP), 20 h corrosion rate (Icorr), and anodic polarization (E-i) curves of 11 commercial Pd alloys were measured in a phosphate buffered saline solution. The alloys were divided into the following four groups based upon composition: PdGa(Ag), PdCu, PdAg, and AuPd and tested in both as-cast and oxidized conditions. In both the as-cast and oxidized conditions, the OCP of Ag-containing Pd alloys is significantly lower than non Ag-containing high-Pd alloys. The OCP of all alloys increased after oxidation. With regard to corrosion rate, the Ag-containing alloys showed a decrease in Icorr with oxidation. In contrast, three of the four non Ag-containing high-Pd (>or=74 wt%) alloys exhibited a higher Icorr. A comparison of the anodic polarization curves showed only the alloys containing larger amounts (>or=16 wt%) of Ag displayed a notable difference between as-cast and oxidized states. Oxidation as required during porcelain-fused-to-metal device preparation alters the electrochemical characteristics of the alloys studied. This alteration may be of importance with regard to their potential for Pd allergy.

Physicochemical basis of the biologic properties of mineral trioxide aggregate.

This study characterized the interactions of mineral trioxide aggregate with a synthetic tissue fluid composed of a neutral phosphate buffer saline solution and root canal dentin in extracted human teeth using inductively coupled plasma-atomic emission spectroscopy, scanning electron microscopy, energy dispersive X-ray analysis, and X-ray diffraction. Mineral trioxide aggregate exposed to synthetic tissue fluid at 37 degrees C released its metallic constituents and produced precipitates with a composition and structure similar to that of hydroxyapatite [Ca10(PO4)6(OH)2-HA]. Endodontically prepared teeth filled with mineral trioxide aggregate and stored in synthetic tissue fluid at 37 degrees C for 2 months produced at the dentin wall an adherent interfacial layer that resembled hydroxyapatite in composition. The authors conclude that Ca, the dominant ion released from mineral trioxide aggregate, reacts with phosphates in synthetic tissue fluid, yielding hydroxyapatite. The dentin-mineral trioxide aggregate interfacial layer results from a similar reaction. The sealing ability, biocompatibility, and dentinogenic activity of mineral trioxide aggregate is attributed to these physicochemical reactions.

Micro-X-ray diffraction observation of nickel-titanium orthodontic wires in simulated oral environment.

A micro-X-ray diffraction (micro-XRD) technique has been employed to determine the phases in two superelastic nickel-titanium orthodontic wires that exhibit shape memory in the oral environment and one superelastic nickel-titanium wire that does not exhibit shape memory in vivo. The micro-XRD analyses were performed over the clinically relevant temperature range of 0-55 degrees C, which corresponds to the ingestion of cold and hot liquids, and both straight and bent (135 degrees ) test samples were analyzed. The results showed that for straight (as-received) test samples, the rhombohedral phase (R-phase) was definitely present in one shape memory wire product and perhaps in the other shape memory wire product, but was apparently absent in the superelastic wire product that did not display shape memory. Martensite was observed in all three wire products after bending. Phase transformations occurred with temperature changes simulating the oral environment for straight test samples of the two shape memory wires, but the micro-XRD pattern changed minimally with temperature for straight test samples of the superelastic wire and for bent test samples of all three wire products. The phase transformations revealed by micro-XRD were consistent with results recently found by temperature-modulated differential scanning calorimetry.

Micro X-ray diffraction study of superelastic nickel-titanium orthodontic wires at different temperatures and stresses.

The phase transformation behavior in three commercial nickel-titanium orthodontic wires having different transformation temperatures was studied by micro X-ray diffraction (micro-XRD). Micro-XRD spectra were obtained at three different included bending angles (135 degrees, 146 degrees and 157 degrees) and three different temperatures (25 degrees C, 37 degrees C and 60 degrees C). The regions analyzed by micro-XRD were within the separate areas of a given wire specimen that experienced only tensile or compressive strain. The intensity ratio (M002/A110) between the 002 peak for martensitic NiTi and the 110 peak for austenitic NiTi was employed as the index to the proportions of the martensite and austenite phases. The ratio of martensite to austenite increased in all three nickel-titanium wires with decreasing included bending angle (greater permanent bending deformation), and was lower within the compression area for all wires at all bending angles than within the tension area. Micro-XRD provides an effective method for quantitative evaluation of the proportions of these two phases in nickel-titanium orthodontic wires, even though considerable preferred crystallographic orientation exists because of the wire drawing process.

Mechanical behavior at different temperatures and stresses for superelastic nickel-titanium orthodontic wires having different transformation temperatures.

OBJECTIVE: The purpose of this study was to investigate the mechanical properties of superelastic nickel-titanium orthodontic wires under controlled stress and temperature. METHODS: Three different superelastic nickel-titanium wires were examined using differential scanning calorimetry (DSC), three-point bending test and micro X-ray diffraction (micro-XRD). The three-point bending test was carried out at constant temperature (23, 37 and 60 degrees C) and stepwise temperature changes (37-60 degrees C and to 37 degrees C) (37-2 degrees C and to 37 degrees C). Five specimens of each wire were tested. Micro-XRD spectra were measured at the tension side of the wire when the temperature changed from 37 to 60 degrees C or 2 degrees C. RESULTS: The load during the stepwise temperature changes (37-2 degrees C and to 37 degrees C) was consistent with that measured at a corresponding constant temperature. The micro XRD spectrum clearly showed that the austenite phase was transformed to martensite phase when the temperature is decreased from 37 to 2 degrees C. In a stepwise temperature change (37-60 degrees C and to 37 degrees C), the load became higher than the original load at each corresponding constant temperature. However, there was no detectable change in the micro-XRD spectrum when the temperature was increased from 37 to 60 degrees C. SIGNIFICANCE: The superelastic nickel-titanium wires exhibited complicated and unexpected mechanical properties under stepwise temperature change. This study shows the possibility of qualitative analysis using micro-XRD to understand mechanical properties of these nickel-titanium wires.

XPS study on the weakest zone in the adhesion structure between resin containing 4-META and precious metal alloys treated with different surface modification methods.

Three precious metal alloys, Type IV gold alloy, 14 K gold alloy, and silver-based alloy, were treated with different surface modifications including a metal primer (VBATDT) application, a SiOx coating method, high-temperature oxidation, modification method with a liquid Ga-Sn alloy, and tin electroplating. Then thin PMMA films were bonded with a resin containing 4-META. Water durability at the adhesion interface was evaluated after water immersion, followed by thermal cycling used liquid nitrogen. The weakest zone at the interface was investigated using XPS only for the Ag-Pd alloy specimens that had been surface-treated with as-polishing, adhesive primer, and the SiOx coating method, since peeling of the PMMA film on the surface of specimens surface-treated by other methods was not observed. Metal elements were detected from the resin side at the adhesion interface. The chemical states of Cu in the resin before argon ion etching were characterized as metal oxides and/or states of chemical interaction with 4-META, VBATDT, or SiOx.

Carbohydrate-carbohydrate binding of ganglioside to integrin alpha(5) modulates alpha(5)beta(1) function.

Gangliosides GT1b and GD3, components of keratinocyte membranes, inhibit keratinocyte adhesion to fibronectin. Although ganglioside sialylation is known to be important, the mechanism of inhibition is unknown. Using purified insect recombinant alpha(5) and beta(1) proteins and alpha(5)beta(1) integrin from lysed keratinocyte-derived SCC12 cells, we have shown that GT1b and GD3 inhibit the binding of alpha(5)beta(1) to fibronectin. Co-immunoprecipitation of GT1b and alpha(5)beta(1) from SCC12 cells and direct binding of GT1b and GD3 to affinity-purified alpha(5)beta(1) from SCC12 cells and insect recombinant alpha(5)beta(1), particularly the alpha(5) subunit, further suggest interaction between ganglioside and alpha(5)beta(1). The carbohydrate moieties of integrin appear to be critical since gangliosides are unable to bind deglycosylated forms of alpha(5)beta(1) from SCC12 and insect cells or poorly glycosylated recombinant alpha(5)beta(1) from Escherichia coli cells. The GT1b-alpha(5)beta(1) interaction is inhibited by concanavalin A, suggesting that GT1b binds to mannose structures in alpha(5)beta(1). The preferential binding of GT1b to high mannose rather than reduced mannose ovalbumin further implicates the binding of GT1b to mannose structures. These data provide evidence that highly sialylated gangliosides regulate alpha(5)beta(1)-mediated adhesion of epithelial cells to fibronectin through carbohydrate-carbohydrate interactions between GT1b and the alpha(5) subunit of alpha(5)beta(1) integrin.

Electrochemical characteristics of high-Pd alloys in relation to Pd-allergy.

OBJECTIVE: The aim of this study was to determine whether the Pd-Cu-based dental ceramic alloys possess any electrochemical characteristics distinguishable from that of other Pd-containing alloys. Of all Pd-containing alloys, this particular alloy group has been linked to frequent incidence of allergy and hypersensitivity reactions. Electrochemical corrosion may instigate these reactions. METHODS: Four groups of alloys, Pd-Cu, Pd-Ga-(with and without Ag), Pd-Ag, and Au-Pd, were evaluated by traditional corrosion measurement techniques in a phosphated buffer saline solution at 20 degrees C. The electrochemical characteristics measured were: (1) 20 h open circuit potential (OCP); (2) 20 h corrosion rate (Icorr); and (3) anodic polarization (E-i) curves. RESULTS: The OCP values (232 +/- 25 mV) of the Ag-free Pd-Ga and Pd-Cu-based alloys were higher than and distinctly different from that (144 +/- 52 mV) of the Ag-containing alloys. The Icorr values of different alloys, despite varied compositions, were indistinguishable from one another. The E-i curves of all alloys were essentially similar, with the Ag-containing (> 5 wt%) alloys showing a subtle difference in their anodic slope within 100 mV above their corrosion potentials. SIGNIFICANCE: The OCP values of Pd-Cu alloys and the Ag-free Pd-Ga alloy are comparable to that reported for pure Pd (239 +/- 21 mV), which indicates that during corrosion these alloys undergo dealloying and consequent Pd-enrichment on their surface. Such a condition is conducive to the release of allergenic Pd++ ions and offers a plausible explanation for the frequent incidence of hypersensitivity reactions associated with the Pd-Cu alloys. The OCP values in other alloys are attributed to dealloying followed by surface enrichment with Ag and/or Au and the possible formation of an insoluble AgCl surface film on the respective alloy surfaces. These events have the potential to suppress or prevent Pd++ ion release. Alloys showing these characteristics have seldom been linked to allergic reactions.

[A case of AIDS complicated with liver tuberculosis].

We experienced a double infection of tuberculosis and amebiasis of the liver. A 28 year old male with AIDS was admitted to our hospital because of severe diarrhea and liver abscess by Entamoeba histolytica. In spite of improvement of the diarrhea and liver abscess by the therapy against E. historicica, serum levels of gamma-GTP and ALP remained high and hepatosplenomegaly gradually increased. A liver biopsy was performed. Pathology showed a granulomatous lesion with Langhans' giant cells. From this specimen, IS6110 gene, a specific DNA for Mycobacterium tuberculosis was detected by PCR method. After anti-tuberculosis treatment was given for 6 months the increased serum gamma-GTP, ALP decreased and hepatosplenomegaly diminished.

Effect of Pd or Au addition on age-hardening in AgMn-based alloys.

OBJECTIVE: The objective of this research was to characterize the age-hardening behavior of AgMn alloys modified with Au or Pd. These alloys are being studied as possible alternatives to Type III dental alloys. METHODS: The age-hardening reactions in Ag-37 at % Mn alloys with 5 at% Au or Pd were investigated by optical microscopy, electrical resistivity, X-ray diffraction and hardness tests. RESULTS: Optical microscopy showed no grain boundary precipitates formed after overaging. The maximum hardness reached by aging in all alloys is about 260 (Hv) and identical to that of traditional AgPdCu dental alloys. The precipitation reactions were retarded with the addition of 5 at% Au or Pd. X-ray diffraction studies suggest a fine precipitate, alpha-Mn, is dispersed within the grain interior. The activation energy for the precipitation reaction is 114 kJ/mol for the AgMn and AgMnAu alloys. This value is about one-half of the activation energy for volume diffusion of manganese in AgMn alloys. SIGNIFICANCE: The age-hardenable AgMn-based alloys modified with Au or Pd can achieve hardness values comparable to those of traditional AgPdCu alloys. Provided the in vivo corrosion resistance of these alloys is adequate, they show promise as a Cu-free alternative material for cast restorations.

Identification of HLA-A3-restricted cytotoxic T lymphocyte epitopes from carcinoembryonic antigen and HER-2/neu by primary in vitro immunization with peptide-pulsed dendritic cells.

The human carcinoembryonic antigen (CEA) and HER-2/neu are potential target antigens for CTL specific immunotherapy for common malignancies such as breast, lung, colon, and gastric carcinomas. Several CTL epitopes restricted by HLA-A2, the most common human histocompatibility molecule, have been previously reported. However, to develop CTL-based immunotherapies for the general population, it is necessary to identify epitopes restricted by other common histocompatibility alleles. Here, we describe two HLA-A3-restricted CTL epitopes from the CEA and HER-2/neu antigens. HLA-A3 binding synthetic peptides from CEA and HER-2/neu were tested for immunogenicity by in vitro primary CTL induction protocol using peripheral blood mononuclear cells from normal healthy volunteers. One peptide from CEA (CEA[9(61)]: HLFGYSWYK) and one peptide from HER-2/neu (HER2[9(754)]: VLRENTSPK) were shown to induce CTL that was capable of killing a tumor cell line expressing HLA-A3 and the corresponding tumor-associated antigen. Additional MHC binding studies with the most common HLA molecules belonging to the HLA-A3 superfamily (HLA-A*1101, -A*3101, -A*3301, and -A*6801), demonstrated that CEA[9(61)] binds five of five A3 supertype molecules with high affinity, and the HER2[9(754)] epitope was able to bind to four of the same five alleles. These results indicate that these two new CTL epitopes should be immunogenic in individuals expressing either HLA-A3, or other members of the HLA-A3 superfamily.

Relationship between Af temperature and load changes in Ni-Ti orthodontic wire under different thermomechanical conditions.

Simple three point bending tests were performed on Ni-Ti wires with three different Af points (1 degree C, 13 degrees C and 34 degrees C) to clarify the relationship between Af temperature and load changes under constant deformation. Each wire was deformed at 37 degrees C and then thermal changes were imposed by temperatures of 2 degrees C or 60 degrees C. The load changes with thermal changes from 37 degrees C to 2 degrees C or 60 degrees C showed the same tendency on the wires with different Af points: In the loading stage, the load became lower than the initial level at 37 degrees C and in the unloading stage, the load became higher than the initial load. The largest load change in the unloading stage was measured with the 13 degrees C Af point wire. Care must be taken when handling Ni-Ti wire with an Af point of less than 1 degree C in order to prevent it from reaching the limit of critical stress of slip deformation when the temperature in the mouth rises to above 40 degrees C.

[A case of bacillary dysentery caused by new quinolone-resistant Shigella flexneri 2a].

A 73-year-old male was admitted to our hospital because of detection of Shigella flexneri 2a from his stool. Antimicrobial treatment with levofloxacin (LVFX) was started, but could not eliminate the organism in the stool. In the examination of drug susceptibility, this strain was highly resistant to all new quinolones. The minimal inhibitory concentration of norfloxacin, ofloxacin and ciprofloxacin to this strain was 12.5 micrograms/ml, 6.25 micrograms/ml and 6.25 micrograms/ml, respectively. The dual mutations were detected in the codon 83 and 87 of the gyrA gene by sequencing the quinolone-resistance determining region (QRDR). There was, however, no significant difference between the intracellular uptake of ciprofloxacin in this strain and in the ciprofloxacin-sensitive strain. The amount of ciprofloxacin in this strain unchanged when carbonyl cyanide m-chlorophenyl hydrazone (CCCP) was added. These results suggest that the advanced resistance in Shigella flexneri against new quinolones could be acquired by only this dual mutations without the change of the active efflux mechanism.

Identification of gp100-derived, melanoma-specific cytotoxic T-lymphocyte epitopes restricted by HLA-A3 supertype molecules by primary in vitro immunization with peptide-pulsed dendritic cells.

The human melanocyte lineage-specific antigen gp100 contains several epitopes recognized by cytotoxic T lymphocytes (CTL). However, most of the epitopes reported to date are HLA-A2.1-restricted. Despite the high frequency of HLA-A2.1 in melanoma patients, effective population coverage requires the identification of epitopes restricted by other frequent HLA alleles. Herein, HLA-A3 binding, gp100-derived synthetic peptides were tested for their capacity to elicit anti-melanoma CTL in vitro using CD8+ T cells from healthy donors as responders and peptide-pulsed autologous dendritic cells as antigen-presenting cells. Of 7 peptides tested, 2 (gp100[9(87)] and gp100[10(86)]) induced CTLs that killed melanoma cell lines expressing HLA-A3 and gp100. Additional MHC-binding studies to various HLA molecules belonging to the HLA-A3 superfamily (HLA-A*1101, -A*3101, -A*3301 and -A*6801) were performed to determine whether these CTL epitopes could further increase potential population coverage. Further experiments indicated that the peptide gp100[9(87)], which bound to HLA-A11 with high affinity, was capable of inducing specific CTLs that killed melanoma cells expressing gp100 and HLA-A11 molecules. Our results indicate that the gp100[9(87)] peptide corresponds to a CTL epitope which may be restricted by either the HLA-A3 or HLA-A11 allele, emphasizing its utility for the design and development of epitope-based therapies for melanoma.

An immunocytochemical technique with monoclonal antibodies to glycosphingolipids in rat primary cerebellar cultures: influence of detergent permeabilization.

Glycosphingolipids (GSLs) are highly expressed in the vertebrate central nervous system. GSLs have been implicated in a variety of phenomena involving cell-cell recognition, neurite outgrowth, synaptogenesis, transmembrane signalling and cell growth and differentiation. We recently determined the distribution of GSLs in rat brain tissues and in primary rat cerebellar cultures as well as using a number of MAbs to GSLs, which were generated and characterized in our laboratory. These results suggested that (i) the expression of GSLs was highly localized to a specific cell type and layer in the rat brain tissues and (ii) some GSLs may be useful markers for identifying cells in the primary cultures. In the present paper, we describe in detail an immunofluorescence technique for the detection of GSL expression in the primary cultures. We demonstrate that the localization of GSLs can be greatly influenced by detergent treatments.

Delineation of the epitope recognized by an antibody specific for N-glycolylneuraminic acid-containing gangliosides.

P3 is a mouse monoclonal antibody (mAb) that binds to several NeuGc-containing gangliosides. It also reacts with antigens expressed in human breast tumors (Vázquez et al. (1995) Hybridoma , 14, 551-556). In this work, the binding specificity of P3 has been characterized in more detail using a panel of glycolipids that included several disialylated gangliosides and several chemical derivatives of NeuGc-GM3. The carboxyl group and the nitrogen function of sialic acid were found to play important roles in the antibody binding, whereas the glycerol tail appears to be nonrelevant. Molecular modeling was used to analyze the binding data, including the finding that P3 selectively recognizes the internal NeuGc in GD3. For this purpose, conformational studies of GD3 were performed using molecular dynamics. It was concluded that sialic acid binds the P3 antibody through its upper face (the one on which the carboxyl group is exposed) and the C4-C5 side of the sugar ring, whereas none or very little contact between the galactose residue and the protein is evident. Conformational analysis of GD3 revealed that, despite the large flexibility of the NeuGcalpha8NeuGc linkage, the P3 binding epitope on the external sialic acid is not well exposed for any of the possible conformations this linkage can adopt, whereas the internal sialic acid presents the epitope in a proper way for several of these conformations. As a final result, a coherent picture of the epitope that fits the wide binding data was obtained.

The multi-epitope approach for immunotherapy for cancer: identification of several CTL epitopes from various tumor-associated antigens expressed on solid epithelial tumors.

One approach to development of specific cancer immunotherapy relies on the induction of cytotoxic T lymphocytes (CTL) specific for tumor-associated antigens (TAA). Induction of TAA-specific CTL could be used towards the eradication of established tumors, or to prevent their dissemination or recurrence after primary treatment. The present study identifies a set of CTL epitopes from TAA frequently found on solid epithelial tumors such as breast, lung and gastro-intestinal tumors. Specifically, HLA-A2.1 binding peptides from the MAGE2, MAGE3, HER-2/neu and CEA antigens were tested for their capacity to elicit in vitro anti-tumor CTL using lymphocytes from normal volunteers and autologous dendritic cells as antigen-presenting cells. A total of 6 new epitopes (MAGE2[10(157)], MAGE3[9(112)], CEA[9(691)], CEA[9(24)], HER2[9(435)] and HER2[9(5)]) were identified which were capable of specifically recognizing tumor cell lines lines expressing HLA-A2.1 and the corresponding TAA. In one case (CEA[9(24)]), induction of vigorous anti-tumor CTL responses required epitope engineering to increase HLA-A2.1 binding affinity. Finally, most of the newly identified epitopes (5 out of 6) were found to be highly crossreactive with other common HLA alleles of the A2 supertype (A2.2, A2.3, A2.6 and A6802), thus demonstrating their potential in providing broad and non-ethnically biased population coverage. The results are discussed in the context of the development of multi-epitope-based therapies with broad applicability for patients suffering from commonly found tumors.

In vitro immunization and expansion of antigen-specific cytotoxic T lymphocytes for adoptive immunotherapy using peptide-pulsed dendritic cells.

The design of an effective procedure to sensitize and expand antigen-specific cytotoxic T lymphocytes (CTL) in vitro is essential for the development of effective adoptive cellular immunotherapy protocols for cancer. We have analyzed the capacity of tissue culture-derived dendritic cells (DC) to present specific peptide epitopes to CTL precursors. Our results demonstrate that peptide-pulsed DC were efficient in generating CTL responses specific for various viral and tumor epitopes. Furthermore, IL-7 and IL-10 potentiated the ability of the peptide-pulsed DC to trigger antigen-specific CTL responses. The CTL generated using this procedure efficiently recognized the naturally processed antigens and could be expanded approximately 100- to 1000-fold in tissue culture in 10 to 15 days without a loss of activity and specificity. The results and procedures described herein may facilitate the development of effective CTL-based adoptive immunotherapy for chronic viral diseases and cancer.