An investigation of the pH-activity relationships of Cex, a family 10 xylanase from Cellulomonas fimi: xylan inhibition and the influence of nitro-substituted aryl-beta-D-xylobiosides on xylanase activity

The kinetic parameters of Cex, a family 10 xylanase from Cellulomonas fimi, were determined at various pH levels using soluble birchwood xylan (BWX) as a natural polymeric substrate along with three other synthetic aryl-beta-D-xylobioside substrates. Using BWX, a high level of substrate inhibition was observed which increased with decreasing pH. In contrast, typical Michaelis-Menten-type profiles were obtained using the three aryl-beta-D-xylobiosides as substrates. The k(cat) values determined using o-nitrophenyl-beta-D-xylobioside did not change as the pH increased, whereas the k(cat) values obtained with BWX, phenyl-beta-D-xylobioside and p-nitrophenyl-beta-D-xylobioside decreased, suggesting that the presence of an ortho nitro group affects the activity displayed by Cex. These differences were not observed with XynB from Clostridium stercorarium F9, a member of the same family of xylanases as Cex. These results indicate that a careful evaluation is required when employing substituted aryl-beta-D-xylobiosides in the characterization of xylanases.     

Preparation and properties of carbon dental implant with fine rahmen surface layer

Blade- and tooth-root-form carbon dental implants were prepared by fixing fine Rahmen surface (FRS) layers of carbon fabric on a carbon fiber-reinforced carbon (CFRC) core. The fixing was successfully performed through the low-temperature deposition of pyrolytic carbon using cis-1,2-dichloroethylene as the raw material. The resultant implants showed flexural and compressive strength more than 150 and 100 MPa, respectively. The outer part of this FRS layer contained many pores ranging from 30 to 300 μm in diameter and 40–75% in volume fraction.New bone was generated in the open pores after being embedded in the femur of a monkey for 6 months. The shear strength of the embedded implant vs bone interface was 15 MPa. On the other hand, the blade-form FRS implant in the mandible of a monkey gave a locking system consisting of “bone-collagen fiber-calcified layer” after two years under a masticating pressure. The latter is very similar to the natural tooth system.     

Use of liposome encapsulated hemoglobin as an oxygen carrier for fetal and adult rat liver cell culture

Engineering liver tissue constructs with sufficient cell mass for transplantation implies culturing large numbers of hepatocytes in a reduced volume; however, providing sufficient oxygen to dense cell cultures is still not feasible using only conventional culture medium. Liposome-encapsulated hemoglobin (LEH), an oxygen-carrying blood substitute originally designed for short-term perfusion, may be a good candidate as an oxygen carrier to cultured liver cells. In this study, we investigated the feasibility of maintaining long term hepatocyte cultures using LEH. Primary fetal and adult rat liver cells were directly exposed to LEH for 6 to 14 days in static culture or in a perfused flat plate bioreactor. The functions and viability of adult rat hepatocytes exposed to LEH were not adversely affected in static monolayer culture and were even improved in the bioreactor. However, some cytotoxicity of LEH was observed with fetal rat liver cells after 4 days of culture. LEH, though a suitable oxygen carrier for long-term culture of mature hepatocytes, is not suitable in its present form for perfusing fetal hepatocyte cultures in direct contact with the liposomes; either the LEH will have to be made less toxic or a more sophisticated bioreactor that prevents the direct contact between hepatocytes and perfusates will have to be designed if fetal cells are to be used for liver tissue engineering.     

The Complete Loss of p53 Expression Uniquely Predicts Worse Prognosis in Colorectal Cancer

p53 immunohistochemistry is considered an accurate surrogate marker reflecting the underlying TP53 mutation status and has utility in tumor diagnostics. In the present study, 269 primary CRCs were immunohistochemically evaluated for p53 expression to assess its utility in diagnostic pathology and prognostication. p53 expression was wild-type in 59 cases (23%), overexpressed in 143 cases (55%), completely lost in 50 cases (19%), and cytoplasmic in 10 cases (4%). p53 immunoreactivity was associated with tumor size (p = 0.0056), mucus production (p = 0.0015), and mismatch repair (MMR) system status (p < 0.0001). Furthermore, among CRCs with wild-type p53 expression, a significantly higher number of cases had decreased CDX2 than those with p53 overexpression (p = 0.012) or complete p53 loss (p = 0.043). In contrast, among CRCs with p53 overexpression, there were significantly fewer ALCAM-positive cases than p53 wild-type cases (p = 0.0045). However, no significant association was detected between p53 immunoreactivity and the “stem-like” immunophenotype defined by CDX2 downregulation and ALCAM-positivity. Multivariate Cox hazards regression analysis identified tubular-forming histology (hazard ratio [HR] = 0.17, p < 0.0001), younger age (HR = 0.52, p = 0.021), and female sex (HR = 0.55, p = 0.046) as potential favorable factors. The analysis also revealed complete p53 loss (HR = 2.16, p = 0.0087), incomplete resection (HR = 2.65, p = 0.0068), and peritoneal metastasis (HR = 5.32, p < 0.0001) as potential independent risk factors for patients with CRC. The sub-cohort survival analyses classified according to chemotherapy after surgery revealed that CRC patients with wild-type p53 expression tended to have better survival than those with overexpression or complete loss after chemotherapy. Thus, immunohistochemistry for p53 could be used for the prognostication and chemotherapy target selection of patients with CRC.     

Fabrication of octacalcium phosphate block through a dissolution-precipitation reaction using a calcium sulphate hemihydrate block as a precursor

Although octacalcium phosphate (OCP) powder and a collagen/gelatin composite demonstrate good potential as bone substitutes, an OCP block has not been fabricated to date. In this study, the feasibility of fabricating an OCP block was evaluated through a dissolution-precipitation reaction using a calcium sulfate hemihydrate (CSH) block as a precursor. When the block was immersed in a phosphate salt solution, its composition changed to that of OCP, while its structure was maintained. The diametral tensile strength (DTS) of the OCP block was 1.0?±?0.2?MPa. The macroporosity and microporosity of the OCP block were 33.4?±?4.5% and, 69.0?±?1.6%, respectively. New bone attached well to the OCP block, and this block was partially replaced by bone 2 weeks after implantation. Four weeks after implantation, the surface of the OCP block was nearly covered with new bone and ~30% of the block was replaced by new bone, while no replacement by bone was observed in the case of a hydroxyapatite (HAp) block used as a control. It is concluded that OCP blocks are potentially suitable for their use as artificial bone substitutes.     

Crystallization process and some properties of novel transparent machinable calcium-mica glass-ceramics

Bulk glass having a calcium-mica composition (Ca_0.5Mg₃AlSi₃O₁₀F₂) is homogeneous glass. The crystallization mechanism of the mica is surface crystallization and transparency is lost completely when crystallization occurs on the surface. In this study, by decreasing SiO₂ and increasing CaO and Al₂O₃ from the chemical composition of Ca_0.5Mg₃AlSi₃O₁₀F₂, and moreover by replacing a small amount of K₂O instead of CaO, the phase separation appears in the glasses. Because of this phase separation, the mica begins to be crystallized not only on the surface but also in the bulk at lower temperatures. Consequently, the novel transparent machinable mica glass-ceramic can be obtained by heating the glasses having the chemical composition of Ca_0.6Mg₃Al_1.2Si_2.8O₁₀F₂ and K_0.01Ca_0.595Mg₃Al_1.2Si_2.8O₁₀F₂. As a larger amount of calcium-mica is separated, the bending strength decreases and the fracture toughness increases. Furthermore, by replacing K⁺ ion instead of Ca²⁺ ion in the interlayer of calcium-mica, the interlayer bonding strength becomes high, resulting in the increase of the bending strength.     

Rapid and Continuous Hydrothermal Synthesis of Boehmite Particles in Subcritical and Supercritical Water

Hydrothermal synthesis of AlOOH particles from an Al(NO₃)₃ aqueous solution was examined in subcritical and supercritical water. Continuous and rapid production of AlOOH fine particles was achieved by mixing a metal salt aqueous solution with preheated water fed from another line. The effects of temperature, pressure, and initial concentration of solution on the particle size, morphology, and crystal structure of the metal (hydrous) oxides were examined. These properties are strongly affected by slight changes in pressure and temperature. This strong effect is attributed to the large changes in the properties of water near its critical point resulting from the slight changes in pressure and temperature. The results suggest the simultaneous control of size, crystal structure, and morphology of the particles by hydrothermal synthesis in supercritical water.     

Aqueous solution properties of a silicone surfactant and its mixed surfactant systems

The aqueous solution properties of a nonionic silicone surfactant of dimethylpolysiloxane and its mixed surfactant systems were studied. It was found that the silicone surfactant has a high surface activity and forms micelles in two steps: premicelles in dilute concentrations and polymolecular micelles above 3.7 × 10⁻⁷ mol dm⁻³. In mixed systems of the silicone surfactant with anionic hydrocarbon or fluorocarbon surfactant, weak intermicellar interactions were found. They are due to electrostatic interaction between hydrophilic groups of the respective micelles. Dye solubilization measurements showed that the solubilized amount of Yellow-OB is greater than predicted by ideal systems. Hydrazo-azo tautomerism is observed in fluorocarbon-silicone surfactant systems, while Yellow-OB is solubilized only in the azo-form in the hydrocarbon-silicone surfactant system.     

In situ time-resolved X-ray diffraction of tobermorite formation in autoclaved aerated concrete: Influence of silica source reactivity and Al addition

The hydrothermal formation of tobermorite during the processing of autoclaved aerated concrete was investigated by in situ X-ray diffraction (XRD) analysis. High-energy X-rays from a synchrotron radiation source in combination with a newly developed autoclave cell and a photon-counting pixel array detector were used.To investigate the effects of the silica source, reactive quartz from chert and less-reactive quartz from quartz sand were used as starting materials. The effect of Al addition on tobermorite formation was also studied. In all cases, C-S-H, hydroxylellestadite and katoite were clearly observed as intermediates.Acceleration of tobermorite formation by Al addition was clearly observed. However, Al addition did not affect the dissolution rate of quartz. Two pathways, via C-S-H and katoite, were also observed in the Al-containing system. These results suggest that the structure of initially formed C-S-H is important for the subsequent tobermorite formation reactions.