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Aims: To determine the effect of low-intensity pulsed ultrasound (LIPUS) on cyclooxygenase-2 (COX-2) expression and related mechanisms by using cultured articular chondrocytes derived from porcine mandibular condyles after treatment with interleukin-1 beta (IL-1?). Methods: Chondrocytes were derived from porcine mandibular condylar cartilage and cultured. The cells were treated with or without 10 ng/mL IL-1?. At the same time, the cells were exposed to LIPUS for 20 minutes. After LIPUS exposure, the conditioned medium was changed to a fresh one without IL-1?, and the cells were incubated for 0 to 24 hours. The effects of LIPUS on IL-1?-treated chondrocytes were examined in terms of the expression of p-integrin ?1, COX-2, and phosphorylated extracellular signal-related kinase (p-ERK) 1/2 by real-time polymerase chain reaction (PCR) and Western blot analyses. Differences in the means among multiple groups were examined by oneway analysis of variance (ANOVA) for all groups at each time point, followed by a Scheffe multiple comparison test as a post-hoc test; Student t test was also used. Results: COX-2 mRNA level was upregulated by the treatment with IL-1? and was suppressed significantly (P < .01) by LIPUS exposure. Furthermore, LIPUS enhanced gene expression and phosphorylation of integrin ?, and it inhibited the expression of p-ERK 1/2. Conclusion: LIPUS exposure inhibited IL-1?-induced COX-2 expression through the integrin ?1 receptor followed by the phosphorylation of ERK 1/2. Despite the restricted duration of its effect, LIPUS is suggested to be a potential candidate as a preventive and auxiliary treatment to suppress the degradation of articular chondrocytes in temporomandibular joint osteoarthritis. Keywords: cyclooxygenase-2 (COX-2), low-intensity pulsed ultrasound (LIPUS), mandibular condylar cartilage, mechanical stress, osteoarthritis (OA)

While large-scale eartuquakes are recently reported all over the world, a huge loss of lives and property damages occur in buildings for which seismic design was not applied. In Japan, many seismic retrofits are developed as a construction method, which raises the safety of the building in case of an earthquake. In recent years, seismic retrofits in Korea have been attracting attention as a construction method, which raises the safety of the building in case of an earthquake. In Korean school facilities, a lot of external seismic retrofit methods of construction are adopted. However, enough consideration may not have been given to the transmission of the force between the existing frame and reinforcement frame. In addition, in order to spread the seismic retrofitting of buildings, it is important that the retrofits are easily constructed. So, in this paper, we confirmed seismic capacity using the disk type metal connector that simplified a general connecting form for the purpose of the seismic design of Korean buildings.

Soft tissue or ligament balancing in total knee arthroplasty is important for ensuring knee joint stability. Correct balancing and appropriate alignment of ligaments extend prosthesis life by preventing unnecessary force fromacting on the prosthesis during routine activities. The current implementation of total knee arthroplasty relies heavily on the subjective “feel” of the surgeon for correct prosthesis implantation onto tibiofemoral components. We developed a force distribution sensing systemto provide quantitative information to surgeons during ligament balancing. The measurement system consists of two main components: two force sensors embedded in trial insert for each condyle and signal acquisition for data processing and force visualization. Sensors were designed and developed using pressure-sensitive conductive rubber that measures changes in resistance in the event of deformation caused by external force. Corresponding voltage measured by circuits is transmitted via an RF transceiver to a computer and visualized as color gradient. Current sensors could measure maximum force of 196.13 N (20 kgf). Results from calibration and experiments on a plastic trial prosthesis indicated that the device has good potential for providing appropriate force distribution information on the knee during total knee arthroplasty procedure.

We evaluated the effects of soil washing with ferric chloride (FeCl3) on cadmium (Cd) concentrations in soil solutions and Cd absorption by two spinach cultivars in pot experiments. Soil samples were collected from washed and unwashed plots of rice paddy fields in Fukuoka and Toyama, Japan. Spinach pot cultivation was performed with the above mentioned soil under oxidative conditions. Before and after spinach cultivation, the Cd content in washed soils was always lower than that in unwashed soils. Soil washing with FeCl3 affected the exchangeable cations (i.e. calcium increased and magnesium decreased). The Cd concentration in the soil solution from washed plot was lower than that in the solution from the unwashed plot throughout the spinach growth period, which was attributed to the exchangeable Cd content in both soils, because the fraction equilibrated with the Cd concentration in the soil solution. The exchangeable cation composition was affected by soil washing, but no significant difference in spinach yield was observed between the washed and unwashed plots. The leaf Cd concentration in the two spinach cultivars was up to 70% lower in the washed soils. This study suggested that soil washing in rice paddy fields with FeCl3 was effective for controlling the Cd absorption risk of upland crops such as spinach. However, some risks remain to clear the CODEX standard (0.2 mg kg) in the Cd content of leaf such as spinach, which has a high Cd absorptive capacity. Discipline: Agricultural environment Additional key words: chemical washing, heavy metal, leaf vegetable, pot experiment, remediation technology

In this study, the high performance anchor bar with the sectional area expanded by the steel tube was proposed for the improvement of shear carrying capacity of the placing joint by outside seismic reinforcement, and the abilities of high performance anchor bar were investigated. In this experiment, three diameters of steel tube with the wall thickness of 2mm（the diameters are 15, 20 and 30mm）and four lengths of steel tube（the lengths are 20, 40, 60 and 60mm）were selected as shown in Table 1 and Figure 2. The specimen, the loading method and measurement method as shown in Figures 1 and 4 were used in this experiment. The results obtained by the experiments can be summarized as follows： (1）The uplift of the placing joint increases linearly with increasing the diameter of steel tube in the range under 20mm, but is little affected by the diameter of steel tube in the range over 20mm, and increases slightly with increasing the embedded depth of steel tube. (2）The shear strength of the placing joint reinforced by the high performance anchor bar increases linearly with the increase of the diameter and embedded depth of the steel tube. (3）In the case that the shear strength of the placing joint by outside seismic reinforcement is determined by the bearing strength of the existing mortar with lower strength as compared to the reinforcing mortar, the effects of the diameter and embedded depth of the steel tube on the shear strength of the placing joint can be explained reasonably by using the analytical model proposed in this study. (4）In order to improve the shear carrying capacity of the high performance anchor bar, it is necessary to use the steel tube more than a certain embedded depth.本研究では、新旧接合面近傍のアンカー筋に鋼管シアキーを固定して断面積を増大させた高性能アンカーの提案とそのせん断抵抗性能を実験的に検討した。その結果、補強部の浮き上がり量は、鋼管シアキーの外径と埋込み深さの増大に従って増大すること、新旧接合部のせん断耐力は、鋼管シアキーの外径と埋込み深さの増大とともに増大すること、既存部モルタルの支圧強度によって決まる新旧接合部のせん断耐力は、本解析モデルによって合理的に説明できること、高性能アンカーのせん断抵抗性能を確保するためには、式（1）に必要せん断耐力を代入して求まる埋込み深さ（l/2）以上の鋼管シアキーを用いる必要があること、などが明らかとなった。

It is well known that enamel matrix proteins play a crucial role in tooth root formation and amelogenesis. Because amelogenin is a major enamel matrix protein, it is assumed that amelogenin also affects the metabolism of cementum. However, the biologic functions of amelogenin in cementoblasts remain unclear. The purpose of this study is to examine the effect of recombinant human full-length amelogenin (rh174) on the proliferation of cultured human cementoblast-like (HCEM) and human periodontal ligament (HPDL) cells.HCEM and HPDL cells were cultured and treated with 100 ng/mL rh174 in the presence or absence of an anti-cluster of differentiation (CD) 63 blocking antibody. Cell proliferation was evaluated using a cell proliferation enzyme-linked immunosorbent assay 5-bromo-2-deoxyuridine kit and quantification of the cell number by 3-(4, 5-dimethylthiazol-2-yl)-5-(3-carboxymethoxyphenyl)-2-(4-sulfophenyl)-2 H-tetrazolium-inner salt assay. The phosphorylation of extracellular signal-regulated kinases (ERK) 1/2 was measured by enzyme-linked immunosorbent assay and Western blot analysis.The proliferation of HCEM and HPDL cells was enhanced significantly (P0.05) by treatment with rh174, and inhibited significantly (P0.05) by the addition of anti-CD63 blocking antibody. In addition, the ratio of phosphorylated ERK1/2 to total ERK1/2 became significantly larger (P0.05) by treatment with rh174, and was reduced significantly by the addition of anti-CD63 blocking antibody in both HCEM and HPDL cells.The results show that rh174 interacts with CD63, and rh174/CD63 interaction activates the ERK1/2 signaling pathway, enhancing the proliferation activities of HCEM and HPDL cells.

We examined the localization and boundary lubricating function of superficial zone protein (SZP) on the surface of mandibular condylar cartilage. Chondrocytes were separated from the surface layer of mandibular condylar cartilage of 6- to 9-month-old female pigs. A cyclic tensile strain of 7% or 21% cell elongation was applied to the cultured chondrocytes. Gene expression levels of cartilage matrix proteins and secretory phospholipase A(2) (sPLA(2)) were quantified by real-time polymerase chain reaction analysis. The friction coefficient of the mandibular condylar surface was measured by a friction tester before and after treatment with 0.1 U/ml sPLA(2). Significantly higher mRNA levels of SZP and type I collagen were found in chondrocytes from the superficial layer than in those in the other layers. The SZP mRNA level was up-regulated by cyclic tensile strain of 7% and 21% cell elongation. Cyclic tensile strain of 21% cell elongation up-regulated the sPLA(2) mRNA level. The friction coefficient of the condylar surface was increased significantly by treatment with sPLA(2). The removal of SZP from the surface layer of mandibular condylar cartilage by sPLA(2) resulted in a significant increase in the friction coefficient on the surface of articular cartilage.

Hyaluronan (HA) plays a crucial role in the lubricating and buffering properties of synovial fluid. The purpose of this study was to examine the effects of interleukin (IL)-1beta on HA degradation in cultured synovial membrane cells. The rabbit synovial membrane cell line HIG-82 was cultured with and without IL-1beta. The amounts of HA of varying molecular weights in the medium were analyzed using high-performance liquid chromatography, the mRNA levels of HA synthase (HAS) and hyaluronidase (HYAL) were analyzed by means of real-time PCR, and HYAL activity was analyzed by HA zymography. The amounts of HA with a molecular weight lower than 300 kDa, and between 300 and 1900 kDa, in the culture medium of HIG-82 cells were significantly higher in the presence of IL-1beta. However, the amount of HA with a molecular weight greater than 1900 kDa was significantly lower in the presence of IL-1beta. Both HAS2 and HAS3 mRNA levels were upregulated by treatment with IL-1beta. So, too, were the levels of HYAL1 and HYAL2 mRNA, which resulted in enhanced HYAL activity. However, HYAL activity was inhibited by transfection of HYAL2-siRNA. Our results suggest that IL-1beta is a crucial factor in the fragmentation of HA in inflammatory joints.

Automatic contouring system using electrostatic capacitance sensor can sense electrostatic capacitance between a detector and patients' side space. So the system can be set it at an arbitrary distance. On the other hand, this system is so complicated that it is difficult to forecast the detector motion. We studied the difference of the distance according to the rotation direction and verified the system accuracy. In addition, we studied it again after software updating. We compared the set distance with the distance between phantom and detector surface at the closest point. Next, we scanned the phantom with relative 90 degree detector position in a clockwise direction (CW) and a counterclockwise direction (CCW). We took a video of the detector trajectories and compared them. Finally, we compared the results before and after updating the software. At the set distance of 10 mm, the distance between phantom and detector surface distance was 12.00 mm. And at the set distance of 50 mm, it was 52.00 mm. Thus we confirmed the accuracy and repeatability of this system. For the rotation orbit, the CCW rotation kept the distance near the set distance. But the CW rotation rotated over a bigger distance than the set distance at the center position over the phantom. After software updating, this difference became small. Using this system, the rotation orbits differed by their rotation direction. However, after updating software, results indicated that selecting an adequate set distance for every patient and examination makes the image optimum.

It is well known that low-molecular weight hyaluronan (HA) is detected in human periodontal tissue under inflammatory conditions. HA oligosaccharide (HAoligo) has been demonstrated to induce matrix metalloproteinase (MMP)-1 expression in dendritic cells and chondrocytes, however, the bioactivities of HAoligo in periodontal ligament (PDL) cells remain unclear. In this study, we investigated the effect of HAoligo on MMP-1 expression in human PDL (HPDL) cells and the mechanisms in terms of the signal transmission. HAoligo was generated and purified from commercial human umbilical cord HA. HPDL cells were isolated from healthy ligaments, and cultured with HAoligo for 0-24h. The expression of MMP-1, tissue inhibitor of MMPs (TIMP)-1 and TIMP-2 was analyzed by real-time PCR and Western blot analyses. Effects of specific inhibitors of p38 mitogen-activated protein kinase (MAPK) activity, MAPK kinase activity and NFkB activity on HAoligo-induced MMP-1 expression in HPDL cells were also investigated by real-time PCR and Western blot analyses. HAoligo remarkably enhanced MMP-1 expression in both mRNA and protein levels, but no effect was shown on the expression of TIMP-1 and TIMP-2 mRNAs. Inhibition of p38MAPK activity decreased the MMP-1 expression. Neither inhibition of NFkB nor MAPKK activity affected the MMP-1 expression. It was suggested that HAoligo induces MMP-1 expression in HPDL cells, and p38MAPK plays a crucial role in signal transduction for MMP-1 inducted by HAoligo.

During endochondral ossification, the production of hyaluronan (HA) is strictly and selectively regulated by chondrocytes, with a temporal peak at the hypertrophic stage. This study was conducted to clarify the effects of HA on expression and activity of runt-related gene 2 (Runx2), a potent transcription factor for chondrocyte differentiation in hypertrophic chondrocytes. Immature chondrocytes from an ATDC5 cell line were cultured and differentiated in DMEM/Ham's F12 with pre-defined supplements. Using real-time PCR, the gene expressions of type II collagen, MMP-13, HAS2, and Runx2 in cultured chondrocytes were analysed from days 0 to 18 of cell differentiation. The activity and expression of Runx2 in hypertrophic chondrocytes were analysed after the treatment with HA oligosaccharide (HAoligo) using AML-3/Runx2 binding, real-time PCR and Western blot analysis. The effects of pre-incubation of anti-CD44 antibody on Runx2 expression were also examined. Expression of type X collagen and Runx2 mRNAs reached a maximum at the terminal differentiation of chondrocytes. The activity and expression of Runx2 was significantly inhibited in hypertrophic chondrocytes treated with HAoligo compared to the untreated controls. High molecular weight-HA did not affect the expression or activity of Runx2. The expression of Runx2 mRNA was significantly decreased in hypertrophic chondrocytes treated with anti-CD44 antibody. These results suggest that HAoligo may affect the terminal differentiation of chondrocytes during the endochondral ossification by inhibiting the expression and activity of Runx2.

The 129Xe NMR spectra in red blood cell (RBC) were investigated in vitro focusing upon temperature dependence and the magnetization transfer (MT) experiment. 129Xe signals were observed at 192 and 201 ppm in the RBC solution, which were assigned to 129Xe dissolved in saline and RBC phases, respectively. A presence of the third peak was evidenced by the MT experiment, in which a pre-saturation around 210–220 ppm caused a decrease in the intensity of the 192-ppm peak. To examine the applicability of the 129Xe NMR spectra index for the detection of physiological conditions, the temperature dependence of the chemical shift was studied precisely and accurately in the RBC solution. The multinuclear internal reference method was adopted to express the chemical shift on an absolute frequency scale. The temperature dependence was proved to be non-linear and approximated by a convex curve for the 192 ppm peak, whereas it was linear for the 201-ppm peak. The temperature coefficient amounted to as much as 0.2 ppm/°C (5 °C/ppm) for the 129Xe signal in RBC. A quite similar coefficient was observed for the separation of 192 and 201 ppm peaks, which was measured simply without the adoption of an absolute frequency scale and will be suited for practical application.

Accepted: 2004-04-06, 資料番号: SA1003288000

Trehalose has been shown to inhibit bone loss in ovariectomized (OVX) female mice and excessive osteoclastogenesis in lipopolysaccharide (LPS)-injected mice. These facts suggest that there may be select physiological functions for trehalose. Since orally administrated trehalose is digested to glucose by a specific digestive enzyme, trehalase, trehalase-free experimental animals are necessary to study the physiological roles of trehalose in detail. Therefore, we generated trehalase-deficient (treh −/−) mice using the gene-targeting procedure, and an oral trehalose tolerance test revealed that while the blood glucose level increased rapidly in wild-type mice, it was unchanged in treh −/− mice. Thus, these results demonstrate that there is no alternative enzyme for trehalase and suggest that the treh −/− mice are a useful tool to analyze mechanisms of trehalose action on osteoclastogenesis, and the physiological functions of trehalose and trehalase.

A Pulsed current pressure sintering (PCPS) method was employed to sinter compacts of gas atomized Al-8%Ni-2%Mn-1%Cu-0.8%Zr-0.7%Ti-0.25%Mg alloy powder which consisted of Al-Al 3 Ni eutectic and α-Al supersaturated solid solution due to high quenching rates. Results are as follows: (1) The sintered compacts had Al 3 Ni precipitates on the boundaries of the initial Al particles. As those precipitates increased, the hardness of the compact decreased. Maximum value of hardness of the compact was HV200 and about 3 at% of Ni atom was excessively dissolved in this compact. (2) The hydrogen content was decreased from 60 to 20ppm by employing the sintering schedule to remove the adsorbed gas on the particle effectively. The tensile strength of the sintered compact was increased to 500 MPa which was comparable with that of an extruded compact.

Size dependency of the relaxation time T 1 was measured for laser-polarized 129 Xe gas encapsulated in different sized cavities made by glass bulbs or gelatin capsules. The use of laser-polarized gas enhances the sensitivity a great deal, making it possible to measure the longer 129 Xe relaxation time in quite a short time. The size dependency is analyzed on the basis of the kinetic theory of gases and a relationship is derived in which the relaxation rate is connected with the square inverse of the diameter of the cavity. Such an analysis provides a novel parameter which denotes the wall effect on the relaxation rate when a gas molecule collides with the surface once in a second. The relaxation time of 129 Xe gas is also dependent on the material which forms the cavity. This dependency is large and the relaxation study using polarized 129 Xe gas is expected to offer important information about the state of the matter of the cavity wall.

Crohn's disease is treated by medication, operation, and alimentary therapy, and among of them the usefulness of enteral nutrition has been pointed out. Not only elemental diet (ED) but also low residue diets have been reported to be useful, however, there are few cases of long-term use of such digested diets. We have successfully managed a postoperative patient with Crohn's disease involving the small intestine and colon by enteral nutrition mainly with digested diets which are rich in lipids, especially medium-chain triglyceride (MCT) at home. The enteral nutrition was started in a remission period after operation for peritonitis due to perforation of the ileum. About three years have elapsed up to now, and normalization and maintenance of inflammatory reaction, disappearance of the subjective symptoms, maintenance of good nutritional status and weight gain were observed. The patient has recovered normal social activities without any recurrence. Low molecular peptides and MCT in the digested diet are excellent in the digestion and absorption in the body. It is thought that this digested diet regimen involving such peptides and MCT might contribute to the good nutritional management in this case.

A case of preoperatively diagnosed early carcinoma of the appendix is reported. A 78-year-old woman was admitted to the hospital complaining of lower abdominal pain. Barium enema revealed an elevated lesion of the cecum, and adenocarcinoma of the appendix was diagnosed by colon fiberscopy. Right hemicolectomy with D3 lymph node disection was subsequently carried out. Pathological examination demonstrated intramucosal adenocarcinoma limited to the mucosa. This case of early cancer of the appendix is the second case diagnosed preoperatively. We must be careful in treating appendicitis, taking the possibility of carcinoma into consideration.

Callus and adventitious root cultures of Cornus capitata were established and the production of tannins and related compounds was determined in various culture conditions (media, plant growth regulators, light, etc.). Calli produced mainly procyanidins such as (+)-catechin and procyanidin B-3 on Murashige and Skoog (MS) and Woody Plant (WP) media. The maximum content of (+)-catechin (0.1%, as dry weight) was observed in the calli cultured on WP medium supplemented with 0.1mg/l 2, 4-dichlorophenoxyacetic acid (2, 4-D) and 0.1mg/l benzyladenine (BA), or 2.0mg/l naphthaleneacetic acid (NAA) and 0.1mg/l BA in the light. In contrast, adventitious roots cultured in MS liquid medium supplemented with 3.0mg/l indoleacetic acid (IAA) produced a high level of gallotannins such as 1, 2, 3, 6-tetra-O-galloyl-β-D-glucose and 1, 2, 3, 4, 6-penta-O-galloyl-β-D-glucose.

Currently, the number of Total Knee Arthroplasty (TKA) in Japan is 70,000. Because it is easy to expect increasing the number due to super-aging society, the clinical outcome of TKA surgery has been important The surgery requires proper soft tissue balance to avoid shortening fatigue life of prosthesis. Thus, a measurement system of force concentration about contacting region between femur component and polyethylene insert contributes to prolong the prosthesis. This study examined the force concentration by using our developed measurement system. This study performed two experiment consists of physical and simulated experiment As a result, our developed system could localize the force concentration point A future work is to apply this measurement system to the cadaveric knee.

A rare case of cavernous hemangioma rupture in the liver following a blunt abdominal trauma is reported. A 42-year-old male was brought to the hospital in shock following a blunt trauma to his right hypochondral region during a softball game. Ultrasonography and computed tomography of the abdomen demonstrated intraabdominal massive bleeding and injury to the right lobe of the liver. The lesion was highly enhanced by contrast medium. Upon laparotomy which was done under a diagnosis of hemorrhagic shock due to the hepatic injury, bleeding about 6000cc in volume was noted. Right lobectomy was carried out and the patient recovered well. Microscopic examination revealed the ruptured cavernous hemangioma, 2×3cm in size. In a review of clinical cases of traumatic or spontaneous rupture of hemangioma, most lesion were more than 5cm in diameter and such small lesion as the patient had was rare. In emergency cases of hepatic rupture, we must consider a possible basal disease of the liver.

The effect of structural factors on the bending strength of nultilayer ceramiccapacitor (MLC) monuted on a printed circuit board has been studied, with specialreference to the termination electrodes of MLC. Capacitance of MLC suddenly decreases duringthe bending test. This decrease is caused by the breakage of the ceramic body whichinitiated at the edge of the termination electrodes. It is also shown that theamount of the solder, the shape of termination electrodes, and the bending strength of ceramicbody are the important factors. FEW analysis supported the results. Furthermore, the fluctuation of the bending strength was affected by the position of MLC on the board.

In order to confirm engraftment of sibling-donor marrow cells or lekuemia relapse from the patient leukemic cells, DNA from nucleated cells in peripheral blood or bone-marrow fluid from patients who had received bone marrow transplantation (BMT) was examined using the MCT118 gene (D1S80) (a variable number of tandem repeats: VNTR) and 6 other genes and 9 restriction enzymes before and after BMT. In 5 of 10 patients (Case 1, 3, 4, 7, and 9) a difference between the DNA from donor cells and recipient cells was confirmed using the MCT118 gene, and engraftment of the donor marrow cells could be demonstrated. In Case 7, not only the engraftment of donor cells but also the leukemia relapse from recipient cells were confirmed using MCT118. Among the remaining 5 cases, Case 5, 8, and 10, the engraftment of donor cells was confirmed using the COLA2A1 gne (12q14.3). In Case 5, and 10, the engraftment could be detected using a gene in the CA2 locus (8g22). In Case 2, the presence of cells from the sibling was identified with the THRB gene (3p24) alone. In Case 6, the presence of cells from the donor could be confirmed using the DMD gene (Xp21.3-21.1) alone. In Case 8, the AK1 gene (9q34.1-34.2) could identified the difference between DNA from donor cells and recipient cells only, but the difference could also be detected using the COLA2A1 gene. In none of the cases, the INS gene (11p15.5) could identify the difference between the DNA from donor and recipient cells. Although only 10 cases were investigated, the findings suggest the use of first the MCT118 gene, then the COLA2A1 gene and the other genes when trying to determine engraftment of donor marrow cells or leukemia relapse after BMT.

On-site soil washing with iron(III) chloride reduces Cd levels in soil, and thus the human health risks caused by Cd in food. However, it may threaten aquatic organisms when soil washing effluent is discharged to open aquatic systems. Therefore, we conducted trial-scale on-site soil washing and ecological risk assessment in Nagano and Niigata prefectures, Japan. The ecological effect of effluent water was investigated by two methods. The first was bioassay using standard aquatic test organisms. Twice-diluted effluent water from the Nagano site and the original effluent water from the Niigata site had no significant effects on green algae, water flea, caddisfly, and fish. The safe dilution rates were estimated as 20 times and 10 times for the Nagano and Niigata sites, respectively, considering an assessment factor of 10. The second method was probabilistic effect analysis using chemical analysis and the species sensitivity distribution concept. The mixture effects of CaCl 2 , Al, Zn, and Mn were considered by applying a response additive model. The safe dilution rates, assessed for a potentially affected fraction of species of 5%, were 7.1 times and 23.6 times for the Nagano and Niigata sites, respectively. The actual dilution rates of effluent water by river water at the Nagano and Niigata sites were 2200–67,000 times and 1300–110,000 times, respectively. These are much larger than the safe dilution rates derived from the two approaches. Consequently, the ecological risk to aquatic organisms of soil washing is evaluated as being below the concern level.

(Z)-2-(5-Amino-1,2,4-thiadiazol-3-yl)-2-[(fluoromethoxy)imino]acetic acid (2), a useful chemical modifier of cephalosporin antibiotics, was efficiently synthesized from 2-[(fluoromethoxy)imino]-1,3-propanedinitrile. The stereochemical structure of 2 was unambiguously determined by X-ray crystallography.

Extracellular matrix degradation in mandibular condylar cartilage is mediated by various cytokines in the temporomandibular joint (TMJ). Interleukin-1 beta (IL-1β) is detected in joint structures with pathologic status, and participates in catabolic action in the extracellular matrix. The purpose of this study was to investigate the effects of IL-1β on cyclooxygenase-2 (COX-2) expression and cartilage metabolism using cultured chondrocytes from mandibular condyle. Articular chondrocytes from the porcine mandibular condylar cartilage around the surface were cultured and treated with 0–10 ng/ml IL-1β or 0–1000 ng/ml prostaglandin (PGE 2 ) for 0–24 h. The mRNA levels of COX-2, MMP-1, -3, and -13 were evaluated by real-time PCR analysis. The protein levels of PGE 2 and MMPs were examined by ELISA and Western blot analysis, respectively. The expression levels of COX-2 and PGE 2 were enhanced by exogenous IL-1β in chondrocytes. The mRNA levels of MMP-1, -3, and -13 were up-regulated by PGE 2 treatment dose-dependently. It is shown that the expression of COX-2/PGE 2 was enhanced by IL-1β in articular chondrocytes from mandibular condyle, and that MMP-1, -3, and -13 were induced by PGE 2 , suggesting that IL-1β-induced COX-2/PGE 2 play a crucial role in catabolic processes of mandibular condylar cartilage under inflammatory conditions.

(Z)-2-(5-Amino-1,2,4-thiadiazol-3-yl)-2-[(fluoromethoxy)imino]acetic acid (2), a useful chemical modifier of cephalosporin antibiotics, was efficiently synthesized from 2-[(fluoromethoxy)imino]-1,3-propanedinitrile. The stereochemical structure of 2 was unambiguously determined by X-ray crystallography.

Amelogenins are enamel matrix proteins that play a crucial role in enamel formation. Recent studies have revealed that amelogenins also have cell signaling properties. Although amelogenins had been described as specific products of ameloblasts, recent research has demonstrated their expression in bone marrow stromal cells. In this study, we examined the effect of recombinant human full-length amelogenin (rh174) on the proliferation of human mesenchymal stem cells (MSCs) derived from bone marrow and characterized the associated changes in intracellular signaling pathways. MSCs were treated with rh174 ranging in dose from 0 to 1,000 ng/ml. Cell proliferative activity was analyzed by bromodeoxyuridine (BrdU) immunoassay. The expression of lysosomal-associated membrane protein 1 (LAMP1), a possible amelogenin receptor, in MSCs was analyzed. Anti-LAMP1 antibody was used to block the binding of rh174 to LAMP1. The MAPK-ERK pathway was examined by Cellular Activation of Signaling ELISA (CASE) kit and western blot analysis. A specific MAPK inhibitor, U0126, was used to block ERK activity. It was shown that rh174 increased the proliferation of MSCs and MAPK-ERK activity. The MSC proliferation and MAPK-ERK activity enhanced by rh174 were reduced by the addition of anti-LAMP1 antibody. Additionally, the increased proliferation of MSCs induced by rh174 was inhibited in the presence of U0126. In conclusion, it is demonstrated that rh174 increases the proliferation of MSCs by interaction with LAMP1 through the MAPK-ERK signaling pathway, indicating the possibility of MSC application to tissue regeneration in the orofacial region.

Osteoarthritis (OA) sometimes occurs as a consequence of repeated microtrauma involved in parafunction, which may lead to microfracture in the subchondral bone. The aim of this in vitro study was to evaluate the effects of subchondral osteoblasts in loading with repeated excessive mechanical stress on the metabolism of overlying chondrocytes. A high-magnitude cyclic tensile stress of 15 kPa (30 cycles min(-1)) was applied to the cultured osteoblasts obtained from porcine mandibular condyles. The chondrocytes in alginate beads were then co-cultured with mechanically stressed or unstressed osteoblasts. Chondrocytes co-cultured with unstressed osteoblasts showed a phenotypic shift to hypertrophic chondrocytes, characterized by decreased expression of type II collagen, aggrecan, Sry-related HMG box (SOX-9), and cartilage oligomeric matrix protein (COMP) genes and increased expression of type X collagen and bone sialoprotein (BSP) genes, suggesting that the co-culture may change the chondrocyte differentiation to some extent. These changes were more distinct in chondrocytes co-cultured with excessively mechanically stressed osteoblasts. After co-culture with stressed osteoblasts, the expressions of matrix metalloproteinase (MMP)1, MMP3 and MMP13 genes were also enhanced and the synthesis of DNA, proteoglycan and collagen were significantly decreased in chondrocytes. These results demonstrate that alterations in cartilage metabolism can be induced by stressed osteoblasts, indicating a possible explanation for the onset and progression of OA.

Effect of additives on the degradation of adhesion strength between thick film conductor and multilayer ceramic substrate is studied with special reference to the microstructure. Degradation of adhesion strength is caused by the solder diffusion which brings about the expansion of Ag conductor and destruction of the interface between Ag conductor and multilayer ceramic substrate during ageing at 150°C.Addition of precious metal such as Pd or Pt is effctive to prevent the degradation of adhesion strength. In the case of Ag conductor, γ-Ag3 Sn is easily formed during ageing. Addition of Pd or Pt suppress the formation of γ-Ag3 Sn. Accicular precipitates such as celsian is most effective in toughening the interfacial strength.

Even the potential method, which seems to be most effective for automatic planning of the collision-free manipulator path among obstacles, uses transformation of real space coordinates of the obstacle to joint angle space, so the analyses become difficult or impossible when the manipulator degree of freedom becomes large. This paper proposes the application of fuzzy reasoning to enable handling of the coordinate of the obstacle in the real space coordinate without transformation to joint angle space in order to solve the above-mentioned difficulties, while basically adopting the potential method. Moreover, this proposal adds a great advantage in that much useful different dimensional information can be used for planning, offering a valuable method to further develop this problem. For example, in addition to the information about the deviation from the target and the proximity to the obstacles, the rate of change of these quantities can be included in the planning. This usefulness was confirmed by simulation.

Hyaluronan (HA) has lubricating and buffering functions in joints. Mechanical load is a regulatory factor of HA metabolism in joints, and HA synthesis by synovial membrane cells is modulated by mechanical load. However, the effects of mechanical load on HA catabolism by hyaluronidase (HYAL) in synovial membrane cells remain unclear. The purpose of this study was to evaluate the effects of cyclic tensile load on the expression and activity of HYAL in synovial membrane cells. A cyclic tensile load of 22.8% cell elongation was applied to cultured rabbit synovial membrane cells for 3 to 48 h with or without cycloheximide. HYAL1 and HYAL2 mRNA levels were evaluated by means of real-time polymerase chain reaction (PCR) analysis. HYAL activity in the cell culture was analyzed by means of HA zymography with or without HYAL2-small interfering (si) RNA. Levels of both HYAL1 and HYAL2 mRNA were up-regulated significantly (p < 0.01) by the cyclic tensile load with or without cycloheximide. HYAL activity was detected in the loaded cell cultures and was suppressed substantially by HYAL2-siRNA. HYAL activity was undetectable in unloaded cell cultures. These results show that a cyclic tensile load induces the expression and activity of HYAL in synovial membrane cells, suggesting that increased HYAL by mechanical load affects HA catabolism in synovial fluid.

Hyaluronan (HA) is a component of the extracellular matrices of cartilage contributing to the structural and functional integrity. HA metabolism is regulated by both anabolic and catabolic processes; however, a great deal more of the detail has been unknown yet. The purpose of this study was to clarify the effect of excessive mechanical load on the expression and activity of hyaluronidase (HYAL) in chondrocytes with a special reference to the expressions of IL-1beta and tumor necrosis factor (TNF)-alpha. A cyclic tensile load of 22.8% cell elongation, regarded as an excessive mechanical stimulus, was applied to cultured rabbit knee articular chondrocytes. HYAL1, HYAL2, IL-1beta, and TNF-alpha mRNA levels were examined by quantitative real-time PCR analysis. The HYAL activity in culture medium was examined by HA zymography. Both HYAL1 and HYAL2 mRNA levels were upregulated significantly by the loading in cultured chondrocytes. HYAL activity was also enhanced as compared with unloaded controls. The IL-1beta mRNA level was upregulated significantly by the loading, and TNF-alpha mRNA level was slightly upregulated. HYAL1 and HYAL2 mRNA levels were upregulated significantly by IL-1beta treatment, resulting in a slight increase in HYAL activity. These results show that the expression of HYAL1 and HYAL2 in articular chondrocytes is enhanced by excessive mechanical stimuli and affected in part by induction of IL-1beta, leading to HA catabolism in articular cartilage.

Osteoarthritis (OA) in the temporomandibular joint (TMJ) is a degenerative disease caused by excessive external loading. Recently, it was reported that the damage in the mineralized subchondral bone caused by traumatic impact-loading is responsible for the initiation and progression of cartilage degeneration. Thus far, we have hypothesized that cytokines released from damaged subchondral bone from impact-loading affect the cartilage catabolism under pathological conditions. An impactor of 200 gw was dropped onto the top of a porcine mandibular condyle. After organ culture for 2 days, we investigated the association between the subchondral bone and cartilage using histological and biochemical experiments. The impact-loading induced the expression of IL-1beta immunohistochemically and prominently up-regulated IL-1alpha and IL-1beta mRNA levels in subchondral bone. We confirmed a significant decrease in type II collagen and aggrecan mRNA expressions in chondrocytes by co-culture with osteoblasts after impact-loading, and significant increase in mRNA and protein expressions of IL-1beta in subchondral osteoblasts from impact-loaded subchondral bone. The mRNA expressions of type II collagen, aggrecan, and type X collagen in chondrocytes were decreased significantly by the co-culture with osteoblasts pre-treated by IL-1beta, -6, and TNF-alpha. Among them, osteoblasts pre-treated by IL-1beta affected chondrocytes most strongly. It was also shown that IL-1beta-treated osteoblasts enhanced the MMP-1 mRNA level most markedly in chondrocytes among the four cytokines. These results suggest that the TMJ subjected to impact-loading can increase directly IL-1beta synthesis in the subchondral region, subsequently altering the metabolism of adjacent cartilage and may eventually resulting in the onset and progression of TMJ-OA.