Soft synthesis of single-crystal silicon monolayer sheets.
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Journal Article↗
Biomedical subjects
Hiroshi Nakamura
Publications and source records attributed to Hiroshi Nakamura.
Optically induced melting of colloidal crystals and their recrystallization.
Colloidal crystals melt by applying focused light of optical tweezers and recrystallize after removing it. The disturbed zone by the light grows radially from the focus point and the ordering starts from the interface with the crystal. Although the larger disturbed zone is observed for the higher power optical tweezers, a master curve is extracted by normalization of the disturbed zone. The temporal changes of the normalized disturbed zone are well described with exponential functions, indicating that the melting and recrystallization process is governed by a simple relaxation mechanism.
Biophysical Phenomena↗
Effects of glucosamine administration on patients with rheumatoid arthritis.
The purpose of this study was to examine whether glucosamine has an antirheumatic effect in a randomized placebo-controlled study. The subjects were 51 rheumatoid arthritis (RA) patients: 25 patients in the glucosamine group and 26 patients in the placebo group. Glucosamine hydrochloride at a daily dose of 1,500 mg and placebo, respectively, were administered for 12 weeks along with conventional medication. While significant improvement was not found in joint counts and in the rate of ACR20 responders, the face scale and a visual analogue scale pain were significantly in favor of the glucosamine group. ESR and CRP levels did not change, but serum MMP-3 levels decreased in the glucosamine group. Results of the patients' self-evaluations and the physicians' global evaluations indicated that the glucosamine treatment produced noticeable improvements in symptoms. Although glucosamine administration had no antirheumatic effect evaluated by conventional measures, it seemed to have some symptomatic effects on RA.
Adult↗
Fibulin-4 is a target of autoimmunity predominantly in patients with osteoarthritis.
Autoimmunity to chondrocyte-producing proteins has been reported in patients with osteoarthritis (OA) as well as in those with rheumatoid arthritis (RA). To answer whether or not OA-specific autoimmunity exist, we performed screening of chondrocyte-producing autoantigens by two-dimensional electrophoresis and Western blotting with each of 20 OA and 20 RA serum samples. We identified an apparently OA-specific autoantigen spot with a molecular mass of 52 kDa and a Isoelectric point of 4.1 as fibulin-4 by mass fingerprinting. By preparing recombinant proteins of fibulin-4, we determined prevalence of the autoantibodies to fibulin-4 in 92 patients with OA, 67 patients with RA, 40 patients with systemic lupus erythematosus, and 43 patients with systemic scleroderma. As a result, the IgG type anti-fibulin-4 autoantibodies were detected in 23.9% of sera from patients with OA, in 8.9% of sera from patients with RA, in 2.5% of sera from patients with systemic lupus erythematosus, and in 9.3% of sera from patients with systemic scleroderma. Furthermore, we immunized DBA/1J, ICR, BALB/c, and C57BL/6 mice with the recombinant fibulin-4 proteins to investigate arthritogenecity of fibulin-4. As a result, mild synovitis was detected in all of the four strains. In addition, we demonstrated expression of fibulin-4 in chondrocytes at both mRNA and protein levels in vivo and in vitro by RT-PCR, Western blotting, and immunohistochemistry. Taken together, fibulin-4, expressed in chondrocytes and recognized as an autoantigen mainly in OA rather than in RA, may play pathogenic roles in OA.
Adult↗
Enhanced production of MMP-1, MMP-3, MMP-13, and RANTES by interaction of chondrocytes with autologous T cells.
It has been reported that T cells and chondrocytes interact through cell surface molecules such as MHC, CD4 or CD8 in osteoarthritis (OA) and T cells are activated. The objective of this study is to investigate the responses of chondrocyte-T cell interaction in terms of metalloprotease (MMP) and chemokine production. Articular cartilage and autologous blood were obtained from patients with OA and fracture who under went prosthetic surgery. Synovial fluid (SF) was collected from OA patients. Isolated chondrocytes were co-cultured with autologous T cells. SF cells were analyzed by immunostaining or Alcian blue staining. The production of MMP-1, MMP-3, MMP-13, and regulated on activation, normal T expressed and secreted (RANTES) was enhanced by direct co-culture compared to indirect co-culture using Transwell. Production ratio of RANTES in OA was significantly higher than non-arthritic samples. CD3 positive mononuclear cells and chondrocyte-like cells were found in SF. Chondrocyte-T cell contact was more adhesive in OA samples. These results showed the production of MMPs and RANTES was enhanced by the interaction and that chondrocyte-T cell contact was possible in vivo.
Aged↗
Close-packed colloidal crystalline arrays composed of silica spheres coated with titania.
Titania coated monodisperse silica spheres have been synthesized and fabricated as a close-packed colloidal crystalline array. We have demonstrated that the coated colloidal sphere can be used to control the peak position of the optical stop band through variation of the coating thickness. The titania coated silica spheres were prepared by the layer-by-layer assembly coating process, which reciprocally laminates the cationic polyelectrolyte and the anionic titania nanosheets on a monodisperse silica spheres, and were sintered to change the titania nanosheets to anatase. The Bragg diffraction peak of the colloidal crystalline array shifted to the long wavelength region with an increase of thickness of the titania layer. Angle-resolved reflection spectra measurements clarified that the red shift was caused by increasing of the refractive index with increase of the thickness of the layer. The current work suggests new possibilities for the creation of advanced colloidal crystalline arrays with tunable optical properties from tailored colloidal spheres.
Journal Article↗
Comparative analysis of gene expression profiles in intact and damaged regions of human osteoarthritic cartilage.
OBJECTIVE: To analyze the differences in gene expression profiles of chondrocytes in intact and damaged regions of cartilage from the same knee joint of patients with osteoarthritis (OA) of the knee. METHODS: We compared messenger RNA expression profiles in regions of intact and damaged cartilage (classified according to the Mankin scale) obtained from patients with knee OA. Five pairs of intact and damaged regions of OA cartilage were evaluated by oligonucleotide array analysis using a double in vitro transcription amplification technique. The microarray data were confirmed by real-time quantitative polymerase chain reaction (PCR) amplification and were compared with previously published data. RESULTS: About 1,500 transcripts, which corresponded to 8% of the expressed transcripts, showed > or = 2-fold differences in expression between the cartilage tissue pairs. Approximately 10% of these transcripts (n = 151) were commonly expressed in the 5 patient samples. Accordingly, 114 genes (35 genes expressed in intact > damaged; 79 genes expressed in intact < damaged) were selected. The expression of some genes related to the wound-healing process, including cell proliferation and interstitial collagen synthesis, was higher in damaged regions than in intact regions, similar to the findings for genes that inhibit matrix degradation. Comparisons of the real-time quantitative PCR data with the previously reported data support the validity of our microarray data. CONCLUSION: Differences between intact and damaged regions of OA cartilage exhibited a similar pattern among the 5 patients examined, indicating the presence of common mechanisms that contribute to cartilage destruction. Elucidation of this mechanism is important for the development of effective treatments for OA.
Cartilage, Articular↗
Catabolic stress induces features of chondrocyte senescence through overexpression of caveolin 1: possible involvement of caveolin 1-induced down-regulation of articular chondrocytes in the pathogenesis of osteoarthritis.
OBJECTIVE: Articular chondrocyte senescence is responsible, at least in part, for the increased incidence of osteoarthritis (OA) with increased age. Recently, it was suggested that caveolin 1, a 21-24-kd membrane protein, participates in premature cellular senescence. Caveolin 1 is the principal structural component of caveolae, vesicular invaginations of the plasma membrane. This study was undertaken to investigate whether the catabolic factors oxidative stress and interleukin-1beta (IL-1beta) induce features of premature senescence of articular chondrocytes through up-regulation of caveolin 1 expression. METHODS: Caveolin 1 expression was investigated in human OA cartilage by real-time polymerase chain reaction and in rat OA cartilage by immunohistologic analysis. We studied whether IL-1beta and H2O2 induce caveolin 1 expression in OA chondrocytes and analyzed the relationship between cellular senescent phenotypes and caveolin 1 expression in human chondrocytes. RESULTS: In human and rat OA articular cartilage, caveolin 1 positivity was associated with cartilage degeneration. Both IL-1beta and H2O2 up-regulated caveolin 1 messenger RNA and protein levels, and both treatments induced marked expression of senescent phenotypes: altered cellular morphology, cell growth arrest, telomere erosion, and specific senescence-associated beta-galactosidase activity. Caveolin 1 overexpression induced p38 MAPK activation and impaired the ability of chondrocytes to produce type II collagen and aggrecan. In contrast, down-regulation of caveolin 1 with antisense oligonucleotide significantly inhibited the features of chondrocyte senescence induced by catabolic factors. Caveolin 1 induction and stresses with both IL-1beta and H2O2 up-regulated p53 and p21 and down-regulated phosphorylated retinoblastoma (Rb), suggesting that the p53/p21/Rb phosphorylation pathway, as well as prolonged p38 MAPK activation, may mediate the features of chondrocyte senescence induced by stress. CONCLUSION: Our findings suggest that IL-1beta and oxidative stress induce features of premature senescence in OA chondrocytes, mediated, at least in part, by stress-induced caveolin 1 expression. This indicates that caveolin 1 plays a role in the pathogenesis of OA via promotion of chondrocyte down-regulation.
Aggrecans↗
Distinct signaling pathways are involved in hypoxia- and IL-1-induced VEGF expression in human articular chondrocytes.
Recent data suggest that vascular endothelial growth factor (VEGF) is involved in the pathogenesis of osteoarthritis (OA). Cartilage is an avascular tissue, leading to a low cartilage O2 level. Thus, in a variety of pathologic or physiologic conditions, VEGF is partly regulated by hypoxic stress. The implications of hypoxia for VEGF expression by OA chondrocytes, however, are not known. We investigated the regulatory system of VEGF in OA chondrocytes under hypoxic conditions. Chondrocytes were obtained from articular cartilage of patients with OA. Cells were cultured and then incubated under hypoxic (95% N2, 5% CO2) or normoxic conditions, with or without interleukin (IL)-1 (10 ng/mL) stimulation. The mitogen activated protein kinase (MAPK) inhibitors were also used. VEGF levels in the culture supernatants were measured using an enzyme-linked immunosorbent assay. Western blot analysis was used to examine the expression of hypoxia inducible factor (HIF)-1alpha. Hypoxia significantly increased VEGF levels (p<0.05). Hypoxia-induced VEGF secretion was abolished by p38MAPK inhibitor, but not by JNK inhibitor. In contrast, IL-1-induced VEGF secretion was blocked by JNK inhibitor, and not by p38MAPK inhibitor. Both hypoxia and IL-1-induced HIF-1alpha were attenuated by p38 MAPK and JNK inhibitors. We demonstrate that hypoxia and IL-1 induce VEGF production in chondrocytes through distinct MAPK signaling pathways, indicating that VEGF is induced in a HIF-1-dependent or -independent manner in chondrocytes.
Aged↗
Effectiveness of dental computed tomography in diagnostic imaging of periradicular lesion of each root of a multirooted tooth: a case report.
Radiography by use of the three-dimensional (3D) Accuitomo XYZ Slice View Tomograph (3DX), a device for compact computed tomography in dentistry, was performed as a new diagnostic imaging technique for a patient who needed endodontic therapy. The 3DX was used for examining and diagnosing the presence and expansion of periradicular lesions in each root of a multirooted tooth. High-resolution 3D images were observed and compared with those obtained by routine conventional radiography. It was difficult to diagnose the cause of clinical symptoms by general examinations including the usual intraoral radiography and panoramic radiography. The images obtained by the 3DX, however, clearly showed the apparent presence and expansion of a periradicular lesion in only one root of the multirooted tooth. The 3DX, a newly developed dental computed tomography, provides abundant new image information not attainable by either intraoral radiography or panoramic radiography, and its application for clinical use is very effective for examining and diagnosing regions of interest for endodontic therapy.
Diagnosis, Computer-Assisted↗
Role of IL-2 and helper T-lymphocytes in limiting periapical pathosis.
The purpose of this study was to examine the role of IL-2 and helper T-lymphocytes in the development of periradicular lesions in rats. In control animals, periradicular lesions developed within 28 days following pulpal infection. Immunologically, some anti IL-2 and anti CD4-reactive helper T-lymphocytes infiltrated the periapical tissue at 14 days, and their numbers increased at 28 days. In experimental animals, tacrolimus (FK506) was injected every day to inhibit the IL-2 production by helper T-cells. Histologically, the pulpal necrosis and periradicular inflammation in tacrolimus-treated rats were more severe than those in the control rats. Furthermore, the areas of pulpal necrosis and periradicular lesion in the immunosuppressed rat were significantly greater than those in the normal ones. The numbers of IL-2- and CD4-positive cells in the lesion of the experimental rats were statistically lower than those of the control ones. These results show that the decrease in IL-2 might have promoted the development of periradicular lesions.
Alveolar Bone Loss↗
Comparison of surgical outcomes between macro discectomy and micro discectomy for lumbar disc herniation: a prospective randomized study with surgery performed by the same spine surgeon.
STUDY DESIGN: A prospective study was conducted on the surgical procedures for lumbar disc herniation. OBJECTIVE: The objective of this study is to investigate the surgical outcomes of different methods when performed by the same surgeon, using a prospective study. BACKGROUND: Macro discectomy is widely known as a common surgical procedure for lumbar disc herniation, while microdiscectomy in place of Caspar technique (the Caspar method) and microendoscopic discectomy by a posterior approach are reported as less invasive surgical methods for this condition. However, there have not been a significant number of prospective studies conducted to compare different surgical procedures for lumbar disc herniation. MATERIALS AND METHODS: The target of our study was a group of 62 patients (male: 43, female: 19) who underwent surgery by macro discectomy (A group) and 57 patients (male: 33, female: 24) who underwent surgery by microdiscectomy in place of Caspar technique (B group). The mean ages at surgery were 34 (14 to 62) years and 41 (18 to 65) years respectively, and the mean duration of follow-up was 2 years and 8 months (12 months to 4 years). For all patients, the surgery was performed by 1 of the authors. The items investigated were the operation time, amount of bleeding, duration of hospitalization, amount of analgesic agent used after surgery, pre- and postoperative scores based on judgment criteria for treatment of lumbar spine disorders established by the Japanese Orthopaedic Association score, visual analog scales (VAS, 0 to 10) for lumbago before surgery and at discharge, VAS for sciatica before surgery and at discharge, perioperative complications, and cases requiring further surgery. RESULTS: There were no significant differences between the 2 surgical procedures in the frequency of use of an analgesic agent after surgery, the pre- and postoperative Japanese Orthopaedic Association scores or postoperative VAS for sciatica. Statistically significant differences were observed in the operation time, amount of bleeding, duration of hospitalization, and postoperative VAS for lumbar pain, but the differences were not large, and may not have been clinically significant. CONCLUSIONS: For herniotomy for lumbar disc herniation, both macro discectomy and microdiscectomy are appropriate, as long as surgeons have mastery of the procedures.
Adolescent↗
Proteomic surveillance of autoantigens in relapsing polychondritis.
Relapsing polychondritis (RP) is a systemic inflammatory disease, in which autoimmunity to cartilage-related components is thought to be involved in its pathogenesis. However, the autoimmune profile in RP has not been studied fully. We therefore investigated autoantibodies/autoantigens in RP comprehensively, by 2-dimensional electrophoresis (2DE), subsequent western blotting (WB) and mass spectrometry, using cell-extracted proteins as the antigen source. As a result, we detected 15 autoantigens on 2DE-WB, and further identified five of them. On average, one RP serum recognized approximately 8 out of the 15 autoantigens. Frequencies of the autoantibodies to the 5 identified antigens of tubulin alpha ubiquitous/6, vimentin, alpha enolase, calreticulin, and colligin-1/-2 were 91%, 46%, 36%, 82%, and 36%, respectively. ELISA using recombinant proteins for them revealed that frequencies of the autoantibodies to tubulin alpha ubiquitous, vimentin, alpha enolase, calreticulin, and colligin-1 were 36%, 64%, 46%, 27%, and 18%, respectively. Our data demonstrated that the autoimmune reaction was not restricted to cartilagerelated components, rather a variety of autoimmune responses occurred in patients with RP, which may be involved in the pathophysiology of RP. In addition, the proteomic approach using cell-extracted proteins would be a powerful way to investigate autoantigens.
Autoantibodies↗
Kruppel-like factor 6 (KLF6) affects the promoter activity of the alpha1-proteinase inhibitor gene.
PURPOSE: Keratoconus is a progressive disease that thins and scars the cornea. In keratoconus corneas, levels of degradative enzymes, including lysosomal acid phosphatase (LAP) and cathepsin B, are elevated, and those of inhibitors alpha1-proteinase inhibitor (alpha1-PI) and alpha2-macroglobulin (alpha2-M) are reduced. The present study explored the possible involvement in keratoconus of Krüppel-like factor 6 (KLF6), a transcription factor previously described to be essential for the integrity of the corneal epithelium. The transcript and proteins level of KLF6 and its action in regulating the genes affected in keratoconus were examined in this study. METHODS: Semiquantitative RT-PCR, Western blot analysis, immunofluorescence and in situ hybridization were used to investigate the expression of KLF6 mRNA and protein in normal and keratoconus corneas. Modulation by KLF6 of the promoter activity of alpha1-PI, LAP, cathepsin B, and alpha2-M genes was studied after transient transfection of KLF6 expression plasmid into corneal epithelial cells using promoter-reporter gene assays. Chromatin immunoprecipitation (ChIP) assays were performed to confirm the interactions between KLF6 and promoters of the genes affected in keratoconus. RESULTS: A global increased expression of the transcription factor KLF6 in terms of mRNAs and proteins was observed in total cornea and/or the epithelium in a substantial number of the keratoconus specimens. The promoter activity of the human alpha1-PI gene was suppressed by expression of KLF6 in corneal epithelial cells. The ChIP assay confirmed a physical interaction between KLF6 and the alpha1-PI promoter. CONCLUSIONS: Transcription factor KLF6 downregulates the alpha1-PI gene in corneal epithelial cells and may thereby be involved in keratoconus.
Adolescent↗
The "off-on" fluorescence detection of alkaline earth metal ions by using 1-naphthylacetanilide derivatives: new cation sensors based on oligo-ethylene oxides connecting 1-naphthylacetanilide at its terminals.
New fluorescent ionophores containing the 1-naphthylacetanilide moiety based on oligo-ethylene oxide (1(4) and 1(5)) have been synthesized, and their photochemical behaviors have been studied. In the absence of a metal ion, the 1-naphthylacetanilide moiety showed a weak fluorescence emission (fluorescence "off state"). However, complexation with Ca(2+) induced a large enhancement effect on the fluorescence intensity of 1(4) and 1(5) from the naphthalene ring (fluorescence "on" state). This behavior can be explained in terms of a similar twisted intramolecular charge-transfer relaxation mechanism. Such "off-on" fluorescence responses of 1(4) and 1(5) provide the digital detection of alkaline earth metal ions.
Journal Article↗
[Bone remodeling and mineral homeostasis].
The discovery of receptor activator of nuclear factor-kappaB ligand (RANKL) elucidates the mechanism of osteoclast differentiation and function regulated by osteoblasts. Osteoprotegerin (OPG), a soluble decoy receptor of RANKL, inhibits both differentiation and function of osteoclasts. OPG-deficient (OPG-/-) mice exhibited severe osteoporosis caused by enhanced osteoclastic bone resorption. Deficiency of OPG in human has been shown to result in juvenile Paget's disease. Blood alkaline phosphatase activity of OPG-/- mice was about four times as high as that of wild-type mice. These results suggest that osteoclastic bone resorption coincidentally induces osteoblastic bone formation by an unknown factor (called coupling factor). Collagen sponge disks containing bone morphogenetic protein-2 (rhBMP-2) were implanted into the dorsal muscle pouches in OPG-/- mice and wild-type mice, and bone mineral density (BMD) of the collagen sponge disks was determined every week for 3 weeks. No significant difference in BMD of the disc was observed between OPG-/- mice and wild-type mice. These results suggest that bone formation is accurately coupled with bone resorption at local sites in OPG-/- mice.
Animals↗
Simultaneous registration with ct-fluoro matching for spinal navigation surgery. A case report.
Computer-assisted surgery, which provides simultaneous, multiplanar images of bone structures, has become widely used. However, registration maneuvering remains time consuming. The objective of this paper is to document the usefulness of CT-fluoro matching for spinal navigation. A spinal navigation system (VECTORVISION compact; Brain LAB, Germany) and a digital imaging system (OEC9800; CATHEX, Tokyo, Japan) were used for CT-fluoro matching in cases of L4/5 and L5/S1 posterior lumbar interbody fusion. A reference array was attached to the L4 spinous process. Preoperative CT images and intraoperative fluoro-shots including L4, L5, and S1 were superimposed on the navigation monitor. Following insertion of L4 screws, a reference array remained to be attached to the L4 spinous process, after which a level definition and pre-registration of L5 and S1 vertebrae were performed and the screwing procedure of L5 and S1 was completed without additional fluro-shots. Registration of three vertebrae was completed without paired-point or surface-matching procedures. The calculation time for the registration in a single vertebra was 30 sec. All pedicle screws were seen to be successfully inserted on postoperative CT images. We performed the navigation surgery by matching the preoperative CT images to the intraoperative fluoro-shots without manual registration. This technique may prove useful in the future for anterior spinal surgery and percutaneous screwing without the need for total exposure of the bone surface.
Bone Screws↗
[BMP and osteoclastogenesis].
Bone morphogenetic protein (BMP)-2 markedly enhanced osteoclast differentiation and survival supported by receptor activator of nuclear factor-kappaB ligand (RANKL). RANKL expressed by osteoblasts is a requirement for osteoclastogenesis, osteoblasts also play important roles in osteoclastogenesis through offering the critical microenvironment for the action of RANKL.
Animals↗