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The calcium-phosphate (CA-P) crystals formed in the extracellular organic matrix synthesized by chicken osteoblasts in cell culture were examined after 30, 40, and 60 days of culture by a number of physical and chemical techniques including chemical analyses, X-ray diffraction, transmission electron microscopy of isolated crystals, and resolution-enhanced Fourier transform infrared spectroscopy. The data reveal that the solid inorganic calcium-phosphate phase consists of a very poorly crystalline apatite, having a low carbonate content and containing acid phosphate groups. The chemical and structural characteristics are generally similar to the apatite crystals found in young newly synthesized bone but there were small but significant differences found. The major significant differences noted were the rate at which maturational changes occurred in the crystals formed in cell culture compared with those noted in vivo and in synthetic carbonate apatite crystals equilibrated with the same cell culture medium, and the persistence of labile groups, especially HPO4(-2) ions during a relatively long period of incubation. Despite extensive chemical efforts to degrade the organic constituents and to disperse the individual crystals isolated from the organic matrix constituents, a large proportion of the crystals were found to be organized in both loosely and densely packed relatively large roughly spherical aggregates. A few of the aggregates were organized in the form of fibrils with the crystals oriented with their c-axes roughly parallel to the long axes of the crystal aggregate. With briefer periods of chemical treatment, larger aggregates of crystals were occasionally observed in which there was a distinct axial periodicity of approximately 70 nm. In such collagen-crystal fragments, the crystals were well-oriented with their c-axis roughly parallel to the long axes of the aggregate similar to the organization and relationships between crystals and collagen fibrils in native bone. Isolated crystals were in the shape of thin plates. At the end of 30 days of culture, many of the crystals were clearly larger than those observed in native chick bone, except for those in the very youngest (7- to 8-day-old) embryos. At the end of 40 and 60 days of culture, the crystal habit remained as thin plates but the crystals were predominantly smaller, similar to those found in older embryo and postnatal chicken bone. The marked tendency of the crystals to form relatively large aggregates that resist dispersion by techniques that readily disperse the crystals of bone, and the presence of a significant number of larger crystals has also been observed in studies of calcified cartilage. Resolution enhanced FTIR spectroscopy revealed the presence of a high concentration of labile phosphate groups, especially after 30 days of culture and just after the plateau of mineralization is reached.

BACKGROUND Alginic acid is comprised of complex polymerized polysaccharides, and can be chemically extracted from seaweed. Alginic acid has an inhibitory effect on histamine release, but its molecular mechanisms are not well understood. OBJECTIVE To investigate the effect of alginic acid on the mast cell-mediated anaphylactic and inflammatory reaction using in vivo and in vitro models and elucidate its molecular mechanisms. MATERIALS AND METHOD The effect of alginic acid on an allergy model was analysed by anaphylaxis, a histidine decarboxylase (HDC) assay, and a histamine assay. Cytokine production was analysed by means of ELISA. Cytokine expression was analysed by means of RT-PCR, and Western blotting. Transcription factor activity was analysed by a luciferase assay and a transcription factor-enzyme linked immunoassay. RESULTS Alginic acid dose dependently inhibited compound 48/80-induced systemic anaphylaxis with doses of 0.25-1 g/kg 1 h (P

The bacterium, Acidovorax avenae ssp. avenae causes several important plant diseases including bacterial stripe of rice, bacterial stalk rot of corn, bacterial leaf blight of oats, and red stripe of sugarcane and millet. Although the organism is seed-borne in rice, no reliable seed assay is available. A semiselective liquid medium based on d-sorbitol and l-pyroglutamic acid (SP medium), and two sets of polymerase chain reaction (PCR) primers were designed for use in a BIOPCR assay for detection of A. avenae ssp. avenae in rice seeds. External primer set, Aaaf3 and Aaar2, and internal primer set, Aaaf5 and Aaar2, designed from a 619 bp fragment of the internal transcribed spacer region of the 16S–23S rDNA of A. avenae ssp. avenae strain CAa4 were specific at the subspecies level. A nested-PCR assay produced the expected DNA product from 58 rice strains tested but not with DNA from 27 strains of A. avenae ssp. avenae from corn and other hosts. Furthermore, the primers failed to amplify a PCR product from two strains of A. avenae ssp. cattleyae, 10 strains of A. avenae ssp. citrulli, or 64 other bacteria. When A. avenae ssp. avenae was enriched prior to PCR by incubating washings of naturally contaminated rice seeds for 12 h in SP liquid medium, populations of the pathogen increased over 1000-fold, whereas populations of saprophytic bacteria remained stable. Populations of the target bacterium routinely reached PCR-detectable levels after seed samples containing as few as 1–2 target cells and up to 10 4 cfu of other bacteria were soaked overnight in phosphate buffer and then enriched for 12 h in SP liquid medium. Assays of 64 naturally infected rice seed samples resulted in 53 being positive by BIO-PCR and 34 by classical PCR. The BIO-PCR assay provides a sensitive, reliable tool for the specific detection of A. avenae ssp. avenae in rice seeds. The use of trade, firm or corporation names in this publication (or page) is for the information and convenience of the reader. Such use does not constitute an official endorsement or approval by the United States Department of Agriculture or the Agricultural Research Service of any product or service to the exclusion of others that may be suitable.

This paper presents a spectral stochastic element free Galerkin method (SSEFGM) for the problems involving a random material property. The random material property and resulting system response quantity are represented by a probabilistic spectral expansion techniques (Karhunen–Loeve expansion and Polynomical Chaos series, respectively) and implemented into the element free Galerkin (EFG) analysis. Numerical solutions in 1D linear elastic problem with random elastic modulus are introduced, and compared with those of Monte Carlo simulation (MCS) so as to provide the validation of the proposed approach. The present SSEFGM approach can produce a probabilistic density distribution as well as a first- and second-order statistical moments (mean and variance) of response quantity by a single calculation, which is distinguished from an iterative MCS. Moreover, the method is based on an element free analysis so that there is no need of nodal connectivities, which usually require more time and labourious task than main calculations. Thus the proposed SSEFGM approach can provide an alternative analysis tool for the problems contains a stochastic material property, and demands complex mesh structures. Copyright © 2004 John Wiley & Sons, Ltd.

In this study we have investigated the behavior of fetal rat osteoblasts, cultured up to 23 days, on a bioactive apatite-wollastonite (AW) glass-ceramic and on the same material on which a carbonated apatite layer had been formed by a biomimetic process (AWa). At the last day of culture, the specific activity of alkaline phosphatase activity, as determined biochemically, was about 30% greater on AWa compared with AW disks. After the cell layers had been scraped off, scanning electron microscopic (SEM) observations of the materials' surfaces revealed that mineralized bone nodules remained attached to both surfaces but in larger amounts on AWa. X-ray microanalysis indicated the presence of calcium (Ca) and phosphorus (P) in the bone tissue throughout the AWa surface and Ca, P, and silicon (Si) on the AW surface. The AW/ and AWa/bone interfaces also were analyzed after fracturing of the disks. The interfacial analysis showed firm bone bonding to the AW and AWa surfaces, confirmed by the X-ray microanalytic mappings. These results indicate the importance of surface composition in supporting differentiation of osteogenic cells and the subsequent apposition of bone matrix, which allows a strong bond of the bioactive materials to the bone. Furthermore, prefabrication of a biologic apatite layer by a method that mimics biomineralization could find application to bone-repairing materials.

An apatite layer was formed on polyethyleneterephthalate (PET) substrates by the following biomimetic process. PET substrates were placed on granular particles of a CaO-SiO2-based glass in simulated body fluid (SBF) with ion concentrations nearly equal to those of human blood plasma to form apatite nuclei on their surfaces (first treatment). They then were soaked in modified SBFs, the ion concentrations of which were changed to give a variation in ionic activity product of apatite (IP), in order to make the apatite nuclei grow (second treatment). The Ca/P atomic ratio and the lattice constant c of the formed apatite decreased from 1.54 to 1.40 and from 6.880 to 6.838 A, respectively, with increasing ion concentrations from 0.75 to 2.00 times those of SBF, that is, with increasing IP from 10−96.6 to 10−91.9. This was attributed to an increase in the concentration of HPO42− ion substituting for the PO43− ion sites, which gave an increase in the Ca2+ in the apatite. Even the apatite formed in 1.00 SBF showed a Ca/P ratio of 1.51 and lattice constants a of 9.432 A and c of 6.870 A. The Ca/P ratio and lattice constant c were smaller and the lattice constant a was larger than those of the bone apatite; its Ca/P ratio and its lattice constants a and c, were 1.65, 9.419 A, and 6.88 A, respectively. This was attributed to the lower content (2.64 wt%) of the CO32− ion substituting for the PO43− ion sites of the apatite compared to that of the bone apatite (5.80 wt%). The lower content of the CO32− ion in the apatite might be caused by the lower concentration of HCO3− ion in 1.00 SBF compared to that in human blood plasma. © 1999 John Wiley & Sons, Inc. J Biomed Mater Res, 46, 228–235, 1999.

An NaOH treatment of pure titanium (Ti) forms a sodium titanate hydrogel surface layer with a smooth graded interface structure to the Ti metal substrate. Subsequent heat treatment at 600 degrees C of the NaOH-treated Ti forms an amorphous sodium titanate surface layer with a smooth graded interface structure similar to the Ti metal substrate. These treated Ti metals both form an apatite surface layer with a smooth graded interface structure to the Ti metal substrates in simulated body fluid (SBF). The smooth graded interface structures give a tight bond of the apatite layer to the substrates. Heat treatment at 800 degrees C of the NaOH-treated Ti forms crystalline sodium titanate and a rutile surface layer with a graded interface structure to the Ti metal substrate, which is intervened by a thick titanium oxide. This substrate forms an apatite layer with a graded interface structure to the Ti metal substrate, which is intervened by a thick titanium oxide in SBF. This irregular graded structure gives a less tight bond of the apatite layer to the substrate.

Our previous study showed that titanium metal forms a bonelike apatite layer on its surface in simulated body fluid when it was subjected to NaOH and heat treatments to form a sodium titanate hydrogel or amorphous sodium titanate surface layer. In the present study, bonding strength of the apatite layer formed on the titanium metals to the substrates were examined under tensile stress, in comparison with those of the apatite layers formed on Bioglass 45S5-type glass, dense sintered hydroxyapatite, and glass-ceramic A-W, which are already clinically used. The NaOH-treated titanium metals showed higher bonding strength of the apatite layer to the substrates, which was maximized by heat treatments at 500 and 600 degrees C, than all the examined bioactive ceramics. It is believed that bioactive metals thus obtained are useful as bone substitutes, even under load-bearing conditions.

There is evidence that infection by Coxsackie viruses can serve as an environmental "trigger" for insulin-dependent diabetes mellitus (IDDM). This background information is summarized briefly, and the data in the IDDM component of Genetic Analysis Workshop 5 (GAW5) concerning serum antibodies to Coxsackie B viruses are described.