Enhancement of macrosphelide-induced apoptosis by mild hyperthermia.

Hyperthermia is a useful adjunct in cancer therapy as it can increase the effectiveness and decrease the toxicity of currently available cancer treatments such as chemotherapy and radiation. In the present study, we investigated whether 41 degrees C hyperthermia (mild HT) for 20 min can enhance macrosphelide (MS5)-induced apoptosis in human lymphoma U937 cells. Our results revealed that, compared with MS5 (5 microM) and mild HT alone, the combined treatment exhibited significant enhancement in apoptosis at 6 h, which was evaluated by observing morphological changes and DNA fragmentation. Marked increase in the reactive oxygen species (ROS) generation was observed immediately after the combined treatment. Significant increase in Fas externalization, caspase-8 and caspase-3 activation, and loss of mitochondrial membrane potential (MMP) was found after the combined treatment compared with MS5 and mild HT alone. Moreover, this combination can also alter the expression of apoptosis-related proteins as evident by the cleavage of Bid and down-regulation of Bcl-2 while no change in the expression of Bax was observed. Furthermore, an immediate rise in the intracellular calcium ion ([Ca(2+)]i) concentration was observed after the combined treatment, which continuously increased in a time-dependent manner. In addition, mild HT treatment alone also increases [Ca(2+)]i concentration without inducing apoptosis. Our data indicate that early increase in ROS generation is mainly responsible for the enhancement of apoptosis after the combined treatment.

MAS-NMR spectroscopy studies in the setting reaction of glass ionomer cements.

OBJECTIVES: The main objective is the characterisation of the setting reaction in glass ionomer cements based on experimental ionomer glasses with different fluorine content and a commercial glass ionomer cement liquid by using 13C CP/MAS-NMR, 29Si, 27Al and 31P MAS-NMR spectroscopy in order to receive information specifically about the cross-linking process. METHODS: Different fluorine containing glass compositions based on 4.5SiO2-3Al2O3-1.5P2O5-(5-z)CaO-zCaF(2) where z=0-3, were mixed with a commercially available polymer liquid to form glass ionomer cements. The cements were subjected to 27Al, 13C CP/MAS, 29Si, and 31P MAS-NMR analysis. RESULTS: The 27Al spectra showed clearly the formation of six-fold coordinate Al(VI), that may crosslink the carboxyl groups in the poly-acid molecules. A shift towards to more positive values of the carboxyl peak in the 13C CP/MAS-NMR spectra showed clearly the proton dissociation of the carboxyl groups. A shift towards more negative values was observed in the 29Si MAS-NMR spectra, suggesting formation of hydrated silica gel and consequently formation of additional Si-O-Si bonds. 31P MAS-NMR spectra also reflected changes in the coordination state around a PO4(3-) tetrahedron. Increasing the fluorine content of the glasses resulted generally in increased reactivity during setting, due to promoting cross-linking and repolymerisation of the silicate phase, followed by clear changes in the MAS-NMR spectra. CONCLUSIONS: The cross-linking process during the setting reaction of glass ionomer cements can be followed by MAS-NMR spectroscopy observing the conversion of Al(IV) to Al(VI). The acid base setting reaction is completed in 1 day and no further significant changes in the MAS-NMR spectra can be observed. Further study is required in order to understand the role of phosphorus.

Release profile of antimicrobial agents from alpha-tricalcium phosphate cement.

A new method for treating carious dentine with alpha-tricalcium phosphate (alpha-TCP) dental cement containing antimicrobial agents has been recently introduced. However, the release behavior of antimicrobial agents from this cement has not yet been clarified. The aim of this study is therefore to examine the release profile of the antimicrobial agents from the alpha-TCP cement. Three kinds of antimicrobial agents (metronidazole, cefaclor and ciprofloxacin) were added to two commercially available alpha-TCP cements (new apatite liner type I and type II). The set cements were then immersed in water at 37 degrees C and the released antimicrobial agents and Ca ion were determined at regular intervals for three months. In addition, scanning electron microscopic observations were conducted before and after immersion for three months. The release profile of the cements containing antimicrobial agents varied depending on the types of antimicrobial agents. The incorporation of antimicrobial agents affected the setting reaction of the cements. The release behavior of the drugs also varied depending on the types of the cements. The differences in the release profile between type I and type II cements reflected the structures and compositions of their matrices.

Composite resin restoration and postoperative sensitivity: clinical follow-up in an undergraduate program.

OBJECTIVES: To analyze the relationship between the cavity depth and liners with postoperative sensitivity of resin composite restorations. METHODS: A clinical follow-up was conducted on 319 resin composite restorations made in the final year of an undergraduate program over a 3-year period. Along with the analyses of cavity type, cavity depth, type of pulpal protection and the materials used, the postoperative sensitivity was also examined on each restoration. RESULTS: Thirty-nine percent of the restorations had no protective layer (Group 1). As the depth of the prepared cavities increased, the restorations received one of the three pulpal protection methods; a calcium hydroxide base (Group 2), glass ionomer cement (Group 3), or protection with a calcium hydroxide base in combination with glass ionomer cement (Group 4). The incidence of postoperative sensitivity showed no significant difference among Groups 1, 2 and 3, but was significantly lower in Group 1 than in Group 4. The restorations made in shallow and medium depth cavities demonstrated significantly less-postoperative sensitivity than those made in deep cavities. The newer generation dentine-bonding agents showed a significantly lower incidence of postoperative sensitivity than the early generation group. CONCLUSIONS: Postoperative sensitivity in resin composite restorations was not related to the absence of protective layers but increased with the depth of cavities restored with the resin composite. The type of dentine-bonding agents could also be responsible for postoperative sensitivity.

Radical scavenging by N-aminoazaaromatics.

N-Aminoazaaromatics were found to react with nitric oxide in the presence of oxygen to afford deaminated products in high yields. The reaction proceeded almost instantaneously in various solvents including water, and one to two equivalent of NO was consumed depending upon the amount of oxygen coexisted, and 1 equivalent of N2O was released in the reaction. In addition, N-aminoazoles were deaminated by potassium superoxide to give parent azoles in good yields. Two equivalents of superoxide was consumed, and about half equivalents of both nitrite and nitrate ion were released. The results demonstrated that N-aminoazoles have ability to protect the biological system against the oxidation promoted by radicals such as nitrogen oxides and superoxide.

Total syntheses of novel cytocidal beta-carboline alkaloids, oxopropalines D and G.

A new type of beta-carboline nucleus, N-methoxymethyl-4-methyl-beta-carboline (4) was synthesized by thermal electrocyclic reaction of a 1-azahexatriene system, involving the indole 2,3-bond. The key compound N-methoxymethyl-1-methoxycarbonyl-4-methyl-beta-carboline (2) was then prepared in a four-step sequence. The total synthesis of oxopropaline G (1e) was achieved from this key compound in four steps. Furthermore, the enantioselective total syntheses of (+)-oxopropaline D (1c) and its enantiomer were also achieved by application of the Sharpless oxidation-procedure in nine steps from 2.

The relationship between water absorption characteristics and the mechanical strength of resin-modified glass-ionomer cements in long-term water storage.

The purpose of this study is to elucidate the water absorption characteristics of resin-modified glass-ionomer cements and to also investigate the relationship between the characteristics and mechanical strength after long-term water storage. The mechanism of water diffusion in these cements is also discussed. Water absorption was measured using a gravimetric analysis for 12 m, while the diffusion coefficient was calculated using Fick's law of diffusion. Water solubility was determined based on the weight of the residue in the immersed water. The compressive and diametral tensile strength were measured at 1, 2, 6, and 12 m. A correlation was observed between the diffusion coefficient and equilibrium water uptake, which thus suggests the water in the cements to diffuse through micro-voids in accordance with the 'Free volumetric theory'. A correlation was seen between the solubility and diffusion coefficient of the cements. The deterioration ratio, defined as the ratio of the strength at 12 m versus that at 1 m, was also calculated. Finally, a negative correlation was observed between the deterioration ratio of the compressive strength and the diffusion coefficients of the cements.

Effect of powder grinding on hydroxyapatite formation in a polymeric calcium phosphate cement prepared from tetracalcium phosphate and poly(methyl vinyl ether-maleic acid).

The primary aim of this study was to determine if cements based on poly(methyl vinyl ether-maleic acid) (PMVE-Ma) and tetracalcium phosphate resulted in hydroxyapatite formation. In addition, the mechanical strength of this type of polymeric calcium phosphate cement was evaluated. Cements were prepared by mixing, in a powder/liquid mass ratio of 3.0, an aqueous solution of PMVE-Ma (mass fraction = 25%) and tetracalcium phosphate powders ground for various periods of time. The tetracalcium phosphate powders and set cements were characterized by means of X-ray powder diffraction and scanning electron microscopy. Mechanical strengths of the cements were tested 24 h after mixing. Prolonged grinding of tetracalcium phosphate powder decreased particle size and/or crystallite size and increased lattice distortion. This enhanced the reactivity of the tetracalcium phosphate powder and elevated the extent of crosslinking between PMVE-Ma molecules, resulting in improved mechanical strength. Hydroxyapatite formation was detected in the cement prepared with the most finely ground tetracalcium phosphate powder. The conversion of residual tetracalcium phosphate particles to more thermodynamically stable hydroxyapatite crystals will reduce the solubility of the polymeric cement and increase its biocompatibility.

Structure of bioactive glass and its application to glass ionomer cement.

We prepared a new glass ionomer cement using bioactive CaO-P2O5-SiO2(-MgO) glass and investigated its setting process using FT-IR and MAS NMR analyses. The compressive strengths of the cements depended on the glass composition and a maximum strength of 33.3 +/- 4.7 MPa was obtained using cement with the glass composition of MgO:4.6, CaO:44.9, SiO2:34.2 and P2O5:16.3% in weight. FT-IR analysis showed that the COOH group in the polyacrylic acid decreased and carboxylate ion (COO-Ca2+) increased after the setting reaction. A broad signal appeared around -82 ppm in 29Si MAS-NMR spectra of the glass and a new signal corresponding to hydrated silica gel formation appeared around -102 and -111 ppm after setting. This suggests that Ca2+ was released from the glass powder to form carboxylate salt and that a degree of polymerization in the silicate network increased. The setting mechanism of the cement was found to be essentially the same as in conventional glass ionomer cement.

Formulation of glutaraldehyde disinfectant for alginate impressions.

The effect of buffer agents incorporated in glutaraldehyde disinfectants on the surface quality of dental models was examined by the measurement of surface roughness, X-ray diffraction analysis and SEM observation. Seven experimental glutaraldehyde disinfectants were prepared using two buffer agents, potassium acetate alone or potassium acetate and sodium hydrogen carbonate in combination. Four kinds of sulfate--zinc, calcium, potassium and magnesium sulfate--were added to these disinfectants in order to accelerate the hydration of calcium sulfate hemihydrate. The impressions treated with the experimental disinfectants for 1 h produced stone surfaces which had significantly lower surface roughness values than those treated with the commercial disinfectants (p < 0.05). The X-ray diffraction analysis and SEM observation showed that these superior surfaces were produced as a result of significant reductions in the amount of residual calcium sulfate hemihydrate. Replacement of buffer agents in commercial glutaraldehyde disinfectants with chemicals such as those studied in the present study will improve the surface quality of dental stone.

Effect of fluoride on apatite formation from Ca4(PO4)2O in 0.1 mol L(-1) KH2PO4.

The effect of fluoride on the hydrolysis of tetracalcium phosphate (TTCP; Ca4(PO4)2O) was investigated in 0.1 mol l(-1)KH2PO4 containing 0-83 mol l(-1) KF. Characterization of the final apatite phase formed by the hydrolysis was made with X-ray diffraction and SEM. The initial pH was between 4.5 and 5.4, depending on the solutions, and the pH rapidly increased and was kept constant between 7.3 and 6.5. An increase in KF concentration tended to lower the pH in the final stage of hydrolysis. The calcium concentration was considerably lower than the phosphorus concentration throughout the reaction. The fluoride concentration decreased shortly after the start of hydrolysis. The hydrolysis of TTCP in 0.1 mol l(-1) KH2PO4 proceeded to form hydroxyapatite via DCPD when the KF concentration was low. The hydrolysis product was a calcium-deficient non-stoichiometric hydroxyapatite with a Ca/P ratio of about 1.5. With an increase in the KF concentration in the 0.1 mol l(-1) KH2PO4 solution, TTCP directly transformed into hydroxyapatite containing F- ions or fluorapatite and with improved crystallinity. The addition of fluoride in the solution initially accelerated the formation of apatite. However, the layer of newly formed apatite adhering to the TTCP particles retarded TTCP dissolution; as a result, hydrolysis was delayed. IR analysis showed that the apatite phase contained HPO2-4 ions in the structure. The formula for the hydrolysis product of TTCP in the presence of fluoride can be expressed as follows: Ca10-x(HPO4)x(PO4)10-x(OH)2-x-yFy.

Polymeric calcium phosphate cements derived from poly(methyl vinyl ether-maleic acid).

OBJECTIVES: The purpose of this study was to assess the feasibility of forming polymeric calcium phosphate cements from a mixed powder of dicalcium phosphate/tetracalcium phosphate or only tetracalcium phosphate and poly(methyl vinyl ether-maleic acid) (PMVE-Ma), and to study their setting reaction. METHODS: The setting reaction process of the polymeric cements was evaluated by mechanical strength tests, infrared spectroscopy and x-ray diffraction analysis and compared with that of a water-setting calcium phosphate cement. The mechanical strength data were analyzed using ANOVA and Scheffé's multiple comparisons test. RESULTS: Cements prepared from the mixed powder and 25-30 wt% aqueous solutions of PMVE-Ma had high mechanical strength after 24 h storage in distilled water at 37 degrees C. The hardening mechanism depended on an acid-base reaction between the carboxyl groups of PMVE-Ma and the mixed powder, especially its tetracalcium phosphate component. The formation of hydroxyapatite in the polymeric calcium phosphate cement was not detected and is apparently inhibited as a result of the competing reaction of PMVE-Ma with the mixed powder. SIGNIFICANCE: The cement-forming reaction was significantly faster than that of a water-setting calcium phosphate cement and slower than that observed with the mixed powder and polyacids such as poly(acrylic acid). The characteristics of the polymeric cements suggest that the materials may be useful in cavity lining or endodontic sealing.

Effect of abietic acid and poly(methyl methacrylate) on the dissolution process of zinc oxide-eugenol cement.

The effects of abietic acid and poly(methyl methacrylate) (PMMA) on the dissolution process of zinc oxide-eugenol cement were studied in 0.01 M acetic acid buffer solution (pH 4). Abietic acid markedly reduced the elution of zinc from the cement, because of the matrix stabilization by the reaction between the acid and zinc oxide. The dissolution process of the cement without additives was controlled by surface reaction. With the cement containing abietic acid or PMMA, the diffusion of eluted species through the cement matrix participated in the dissolution process.

[Studies on as-triazine derivatives. XIX. Synthesis of 2,3-diarylpyrazine 2,3-diarylpyridine derivatives as blood platelet aggregation inhibitors].

4,5-Diphenyl-2-ethoxypyrimidine (1), 3,4-diphenyl-6-ethoxypyridazine (2) and 2,3-diphenyl-5-ethoxypyrazine (3) were evaluated for inhibitory activity towards arachidonic acid-induced aggregation of rabbit blood platelet in vitro. 2,3-Diphenyl-5-ethoxypyrazine (3) exhibited significant inhibitory activity. Thus, various 5-substituted 2,3-bis(4-methoxyphenyl)pyrazines were synthesized by the nucleophilic substitution reaction of 5-chloro-2,3-bis(4-methoxyphenyl)pyrazine (9). In a similar manner, substituted 2,3-bis(4-methoxyphenyl)pyridines were prepared from 2,3-bis(4-methoxyphenyl)-6-methylsulfonylpyridine (17), which was synthesized by the cycloaddition retro Diels-Alder reaction of 5,6-bis(4-methoxyphenyl)-3-methylsulfonyl-1,2,4-triazine (16) with norbornadiene. Among the compounds prepared, 6-isopropoxy-2,3-bis(4-methoxyphenyl)-pyrazine (10f) showed the most potent inhibitory activity, which was more than the activity of anitrazafen[5,6-bis(4-methoxyphenyl)-3-methyl-1,2,4-triazine.

Stability constants of Ca-alkyl salicylate complexes and their effect on the dissolution of calcium hydroxide dental cements.

The proton-ligand stability constants (pKa) and Ca-ligand stability constants (log beta) of the five types of alkyl salicylate were determined and the dissolution of the calcium hydroxide cements prepared from those salicylates was examined in water. The dissolution of the cements containing monoesters decreased with the increase in their pKa and log beta values. The cement containing diester showed lower dissolution than that estimated from those values. This seems to be related to the polymeric structure of the cement matrix consisting of the complex of the diester with calcium.

Erosion process of calcium hydroxide cements in water.

Erosion process of calcium hydroxide cements was examined for 7 days by chemical analysis of eluates and observation of structural change of the eroded cements when the cements were immersed in water at 37 degrees C. The elution of salicylate from the set cements as well as that of Ca continued during the immersion time. In the early stage of the erosion, unreacted Ca(OH)2 was preferentially extracted from the cements in comparison with Ca-alkyl salicylate chelate of the cement matrix. The elution rate of N-ethyl o- and p-toluene sulphonamides, which were contained as plasticizer, was higher than those of Ca and salicylate. Insoluble inorganic filler remained at the surface of the cements after extraction of unreacted Ca(OH)2 and disintegration of the cement matrix. Types of salicylate ester seemed to affect the cement durability.

Continued blood cell formation in spherical bodies in a long-term mouse spleen culture.

During the primary culture of spleen fragments of newborn mice, a spherical body (d = circa 200 to 300 microns) as a three-dimensional cellular organization was formed. Continued production of blood cells from the spherical body was observed without changing its size for about 2 months of culture. Without growth factor, the spherical bodies produced mainly lymphocytes and macrophages. Addition of interleukin-3 enhanced their granulocyte formation, and this enhancement was observed even after a prolonged maintenance without growth factors. The spherical bodies were composed of a uniform mixture of endothelial cells and fibroblasts within the body, and cell-cell contacts between lymphocytes and fibroblasts were notable in the periphery. With prolonged culture, the spherical bodies showed a definite change in their structure by sorting two cell types and the blood cell production gradually decreased. These results suggested that a three-dimensional structure was required for the maintenance, growth, and differentiation of blood cell progenitors in the long-term spleen culture.

Tumor-promoting phorbol ester induces alterations of sialidase and sialyltransferase activities of JB6 cells.

Sialidase and sialyltransferase activities were studied in JB6 mouse epidermal cells before and after exposure to phorbol ester, 12-O-tetradecanoyl phorbol-13-acetate (TPA), which irreversibly induces anchorage-independent growth and tumorigenicity. JB6 cells exhibited sialidase activities toward 4-methylumbelliferyl-alpha-D-N-acetylneuraminic acid (4MU-NeuAc) and gangliosides at pH 4.5 in the particulate fraction but apparently not in the cytosol at pH 4.5 or 6.0. In JB6 cells exposed to TPA and in the anchorage-independent transformants, the sialidase activity toward 4MU-NeuAc was decreased and the activity toward gangliosides was increased compared with those in untreated JB6 cells. Immunological analysis with antisera against membrane-associated sialidases I and II revealed that plasma membrane-associated sialidase I was increased and lysosomal membrane-associated sialidase II was decreased under these conditions. TPA treatment also affected the sialyltransferase activities of JB6 cells: and elevation of the transfer activities toward asialo-orosomucoid and asialo-porcine submaxillary mucin but a reduction of GM3 and GD3 synthase activities were observed on exposure to TPA and in cells transformed by TPA to retain anchorage-independency. These results suggest that an increase in sialic acid bound to glycoproteins and a decrease in that bound to glycolipids may occur in JB6 cells exposed to TPA and in the anchorage-independent transformants.

Dissolution mechanism of zinc phosphate dental cement in acetic and lactic acid buffers.

When zinc phosphate cement was immersed in 0.1 M acetic and lactic acid buffer solutions adjusted to pH 4.1, its dissolution rate was determined by chemical analysis. The eroded surface of the cement was examined by X-ray diffraction and SEM observation. The dissolution process of the cement immersed in the acetic acid buffer solution was mainly controlled by the diffusion of releasing species through the cement matrix and that of the cement immersed in the lactic acid buffer solution by the decomposition of the cement matrix at the surface.

Spleen stromal cell lines selectively support erythroid colony formation.

Mouse stromal cell lines (MSS lines) have been established from the spleens of newborn mice in culture at a low serum concentration. These MSS lines support the proliferation and differentiation of the erythroid progenitor cells from mouse fetal livers and bone marrow in a semisolid medium in the presence of erythropoietin. Larger colonies of over 1,000 benzidine-positive erythroid cells were developed from the fetal liver cells on the MSS cell layers after 6 days of incubation. These layers also support the maturation of the erythroid cells since the enucleation process of the latter was observed in large erythroid colonies. Metabolically active MSS cells are apparently required to support the proliferation and differentiation of the erythroid progenitor cells, because neither the MSS cells inactivated with fixation nor the conditioned media of MSS cells promoted the erythroid colony formation. These studies demonstrate that MSS lines specifically support the proliferation and differentiation of the erythroid progenitor cells in vitro and that stroma cells may have a critical function in blood formation in the mouse spleen.