Human osteoclast-like cells are formed from peripheral blood mononuclear cells in a coculture with SaOS-2 cells transfected with the parathyroid hormone (PTH)/PTH-related protein receptor gene

Subclones of the human osteosarcoma cell line SaOS-2 were established by transfecting with an expression vector containing the human PTH/PTH-related protein (PTHrP) receptor, and their abilities to support osteoclast-like multinucleated cell (OCL) formation were examined in coculture with mouse or human hemopoietic cells. Of four subclones examined, SaOS-2/4 and SaOS-4/3 bound high levels of [125I]-PTH and produced a significant amount of cAMP in response to PTH. OCLs were formed in response to PTH in the cocultures of mouse bone marrow cells with either SaOS-2/4 cells or SaOS-4/3 cells. Human OCLs were also formed in response to PTH in the coculture of SaOS-4/3 cells and human peripheral blood mononuclear cells. Adding dexamethasone together with PTH greatly enhanced PTH-induced human OCL formation. Like mouse OCLs, human OCLs formed in response to PTH were tartrate-resistant acid phosphatase positive, expressed abundant calcitonin receptors and vitronectin receptors, and formed resorption pits on dentine slices. Other osteotropic factors such as 1alpha,25-dihydroxyvitamin D3, prostaglandin E2, and interleukin 6 plus soluble interleukin 6 receptors failed to induce mouse and human OCLs in cocultures with SaOS-4/3 cells. Both mouse and human OCL formation supported by SaOS-4/3 cells were inhibited by either adding an antibody against macrophage-colony stimulating factor or adding granulocyte/macrophage-colony stimulating factor. Thus, it is likely that human and mouse OCL formation supported by SaOS-4/3 cells are similarly regulated. These results indicate that the target cells of PTH for inducing osteoclast formation are osteoblast/stromal cells but not osteoclast progenitor cells in the coculture. This coculture model will be useful for investigating the abnormalities ofosteoclast differentiation and function in human metabolic bone diseases.     

Structural Changes of Sol–Gel-Derived TiO2–SiO2 Coatings in an Environment of High Temperature and High Humidity

Structural changes in sol–gel-derived TiO₂–SiO₂ coatings were found to proceed in an environment of high temperature and high humidity as follows: (1) dissociation of Si–O–Ti bonds in the coating by the attack of water vapor, (2) formation of Ti–O–Ti bonds, and (3) nucleation and growth of anatase TiO₂. The coating obtained with the addition of poly(ethylene glycol), PEG, reacts with water vapor more easily than the coating obtained without PEG, since the former is more porous than the latter due to the decomposition of PEG during heat treatment.     

Photoreactivation of Legionella pneumophila after inactivation by low- or medium-pressure ultraviolet lamp

Photoreactivation of Legionella pneumophila after the inactivation by low-pressure (LP) or medium-pressure (MP) UV lamp was investigated in comparison with that of Escherichia coli. An endonuclease sensitive site (ESS) assay was used to determine the number of UV-induced pyrimidine dimers in the genome DNA of L. pneumophila or E. coli, while the survival ratio of each bacterium was also investigated by cultivation methods. L. pneumophila performed photoreactivation with almost complete repair of pyrimidine dimers associated with the quick recovery of survival ratio. A 3 log inactivation of L. pneumophila by LP or MP UV lamp was, respectively, resulted in 0.5 log or 0.4 log inactivation when photoreactivation was completed. Interestingly, L. pneumophila performed equivalent photoreactivation after LP and MP UV lamp exposures while photoreactivation of E. coli was significantly repressed after the inactivation by MP UV lamp. This study indicated that an attention would be required to design and operate a UV disinfection system targeting L. pneumophila. It was further implied that E. coli would not correctly indicate the fate of L. pneumophila in UV disinfection systems when photoreactivation takes place.     

A SKILL-FORMATION MODEL FOR GRINDING OPERATIONS

A good part of grinding operations is still dependent on skilled operators. Against such a background, this study takes up cylindrical plunge grinding, and presents a skill-formation model based on the learning process similar to that in which a grinding operator acquires skills through trial and error. The model repeats learning by giving only the total grinding stock and initial infeed rate, changing infeed rate, and evaluating the size and the removal rate in the process end. The simulation result has proved that the model shows a skill-formation process similar to that of an operator and generates desirable infeed processes.     

Determination of hymexazol in agricultural products by GC-NPD

A method for the determination of hymexazol in agricultural produdcts by gas chromatography with a highly sensitive nitrogen-phosphorus detector (GC-NPD) was investigated. Hymexazol was extracted with acetonitrile, and the acetonitrile layer was separated by salting-out. The water layer was loaded onto a Chem-Elut column. Hymexazol in the water layer was adsorbed on the column, and eluted with ethyl acetate. The acetonitrile layer and the eluate were mixed and evaporated. The residue was dissolved in ethyl acetate, and the sample solution was cleaned up on a C18 column. Hymexazol in the eluate was analyzed by GC-NPD with a high-polarity capillary column (DB-FFAP) and highly deactivated inlet liner.Recoveries of hymexazol spiked in agricultural products (tomato, lemon, soybean and other samples) at the level of 0.1 mug/g ranged from 65.0 to 84.7%. The limit of detection was 0.02microg/g.     

Screening assay of residual antibiotics in livestock samples by LC-MS/MS

A LC-MS/MS screening assay of multi-class antibiotics was developed for 19 residual antibiotics in livestock samples. Sample preparation employed the QuEChERS (Quick, Easy, Cheap, Effective, Rugged and Safe) approach using 0.5% formic acid in acetonitrile-methanol (8 : 2), with salting-out using magnesium sulfate, trisodium citrate and sodium chloride. Recovery values from 5 different livestock samples ranged from 45.5 to 121.6%, and the RSDs were under 18% at two concentration levels. The limit of quantification values of 19 analytes were under 10 μg/kg in all livestock samples, and the procedure can detect almost all analytes under the MRL. Screening capability was confirmed by employing spiked samples. This new screening assay for residual antibiotics in livestock samples is expected to be useful for routine laboratory tests.     

The so-called alkali-carbonate reaction (ACR) — Its mineralogical and geochemical details, with special reference to ASR

Typical examples of so-called alkali-carbonate reaction (ACR) in the Canadian field concretes in Ontario, CSA concrete prism, RILEM concrete microbars and RILEM mortar bar containing Pittsburg aggregate, were examined petrographically based on polarizing microscopy, SEM observation and quantitative SEM-EDS analysis of the reaction products. It was revealed that ASR gel was the main product responsible for the crack formation in concretes, and that this gel had a common nature to that in the typical ASR. That is, ASR gel presented distinctive compositional trend lines, passing from low-Ca ASR gel at [Ca/Si] = 1/2-1/6, [Ca]/[Na + K] = 1.0 to the “convergent point” with [Ca/Si] = 1.3-1.8, [Ca]/[Na + K] = 100 at which chemical equilibrium is attained with CSH gel. The so-called ACR is a combination of deleteriously expansive alkali-silica reaction (ASR) of cryptocrystalline quartz, and harmless dedolomitization which produces brucite and carbonate halo. In laboratory specimens, fine dolomitic aggregate undergoes dedolomitization, and brucite and ASR gel react to form non-expansive Mg-silicate gel on the dolomite crystals. This explains why the mortar bar produces smaller expansion than the concrete microbar, and why the reaction products are so minute that they escape attention by optical microscopy. As a crystalline counterpart, mountainite is a candidate for low-Ca ASR gel, while sepiolite is one for Mg-silicate gel. Concealed ASR was detected in ACR-affected field concretes undergoing ingress of deicing salt which formed Friedel's salt and Cl-doped CSH gel. Compositions of ASR products, methods of sample preparation and analysis for correct identification of ACR, and artifacts were critically reviewed.     

Endo‐α‐Mannosidase‐Catalyzed Transglycosylation

In order for facilitating the synthesis of oligosaccharides, transglycosylation reactions mediated by glycoside hydrolases have been studied in various contexts. In this study, we examined the transglycosylating activity of a Golgi endo ‐ α‐mannosidase. We prepared various glycosyl donors and acceptors, and recombinant human Golgi endo‐α‐mannosidase and its various mutants were expressed. The enzyme was able to mediate transglycosylation from α‐glycosyl‐fluorides. Systematic screening of various point mutants revealed that the E407D mutant had excellent transglycosylation activity and extremely low hydrolytic activity. Substrate specificity analysis revealed that minimum motif required for glycosyl acceptor is Manα1– 2Man. The synthetic utility of the enzyme was demonstrated by generation of a high‐mannose‐type undecasaccharide (Glc₁Man₉GlcNAc₂).     

Alteration of Oral and Perioral Soft Tissue in Mice following Incisor Tooth Extraction

Oral and perioral soft tissues cooperate with other oral and pharyngeal organs to facilitate mastication and swallowing. It is essential for these tissues to maintain their morphology for efficient function. Recently, it was reported that the morphology of oral and perioral soft tissue can be altered by aging or orthodontic treatment. However, it remains unclear whether tooth loss can alter these tissues’ morphology. This study examined whether tooth loss could alter lip morphology. First, an analysis of human anatomy suggested that tooth loss altered lip morphology. Next, a murine model of tooth loss was established by extracting an incisor; micro-computed tomography revealed that a new bone replaced the extraction socket. Body weight was significantly lower in the tooth loss (UH) group than in the non-extraction control (NH) group. The upper lip showed a greater degree of morphological variation in the UH group. Proteomic analysis and immunohistochemical staining of the upper lip illustrated that S100A8/9 expression was higher in the UH group, suggesting that tooth loss induced lip inflammation. Finally, soft-diet feeding improved lip deformity associated with tooth loss, but not inflammation. Therefore, soft-diet feeding is essential for preventing lip morphological changes after tooth loss.     

Effect of Progranulin on Proliferation and Differentiation of Neural Stem/Progenitor Cells after Oxygen/Glucose Deprivation

We previously demonstrated that sivelestat, a selective neutrophil elastase inhibitor, attenuates the cleavage of progranulin (PGRN) and ischemia-induced cell injury in the brain. To obtain further insight into the role of PGRN, in the present study we evaluated the direct effects of sivelestat and recombinant PGRN (rPGRN) on the proliferation and differentiation of neural stem cells in cultures of neural stem/progenitor cells (NS/PC) under the ischemic condition in vitro. We demonstrated that oxygen/glucose deprivation (OGD)-induced cell proliferation of NS/PC was increased by rPGRN treatment. In addition, this increase was accompanied by increased phosphorylation of Akt and GSK-3β (Ser9) after OGD. But none of these responses occurred by treatment with sivelestat. Therefore, activation of the Akt/GSK-3β pathway could well be involved in this proliferative effect of rPGRN. Although OGD and reoxygenation-induced changes in the differentiation of NS/PC into neurons or astrocytes was not affected by treatment with rPGRN or sivelestat, it is noteworthy that rPGRN enhanced neurite outgrowth of β3-tubulin-positive neurons that had differentiated from the NS/PC. These findings suggest that enhancement of proliferation of endogenous NS/PC and neurite outgrowth of differentiated neurons from NS/PC by PGRN could be useful for a new therapeutic approach for cerebral ischemia.