Biomedical subjects

Takashi Nakamura

Publications and source records attributed to Takashi Nakamura.

At least 19 recordsLinked to original sources

PubMed2026

Hearing loss associated with CDC42 in mice and humans (Takenouchi-Kosaki syndrome): CDC42 and RHOQ synergistically function in cochlear hair cells.

CDC42 is involved in multiple signaling pathways, including actin organization and polarity. We previously reported progressive sensorineural hearing loss (SNHL) in inner ear hair cell (HC)-specific Cdc42-knockout (Atoh1-Cre+/-;Cdc42flox/flox) mice. However, the phenotype was milder than expected, suggesting possible redundancy with other Rho-family GTPases. Thereafter, Takenouchi-Kosaki syndrome (TKS), caused by de novo CDC42 mutations and manifesting as SNHL, was reported, in which the p.Y64C mutation was speculated to be constitutively active. However, the relationship between CDC42 status and hearing phenotypes in TKS remains unclear. Using cell models, mouse models, and patient data, we propose that impaired and/or dysregulated cycling between GDP/inactive and GTP/active forms, through either loss-of-function or constitutive activation, can lead to SNHL. Furthermore, to test redundancy, we generated HC-specific Cdc42;RhoQ double-knockout (Atoh1-Cre+/-;Cdc42flox/flox;RhoQflox/flox) mice, which revealed synergistic roles of CDC42 and RHOQ in cochlear HCs. Supporting this synergy, MDCK cells with CDC42 and RHOQ double knockdown showed greater phospho-cofilin, a key regulator of actin turnover, elevation than single knockdowns.

CDC42↗

PubMed2007

The effect of surface incongruity of grafted plugs in osteochondral grafting: a report of five cases.

Although grafted osteochondral plugs should ideally have a smooth surface for mosaicplasty, surface incongruity is sometimes evident at the time of surgery. There may be no problem if there is depression of the grafted plugs, but graft protuberance may have an adverse effect. We studied five knees in five patients who had incongruity (protuberance or depression) of grafted osteochondral plugs at the time of mosaicplasty. The mean age at surgery was 36.6 years (range, 15-65 years), and the mean follow-up period was 32.9 months (range, 24-49 months). All patients underwent second-look arthroscopy after a mean post-surgical period of 14.8 months (range, 3-18 months). We divided the cases so that there were two in the protuberant group (P) and three in the depressed group (D). In P, all patients had a catching sensation about 4 months after surgery, and sometimes pain in the knee joint. Second-look arthroscopy revealed fissuring of the plugs and fibrillation around the recipient site. In D, there were no symptoms due to the depressed plugs. Second-look arthroscopy showed that the depressed areas were covered with fibrocartilage-like tissue, and that the joint surface was smooth. In conclusion, our clinical results and second-look arthroscopic evaluation suggest that isolated osteochondral plug depressions of not greater than 1 mm could still promote acceptable cartilage healing leading to good clinical outcomes. However, plug protuberance at mosaicplasty should always be avoided.

Adolescent↗

PubMed2007

Local application of recombinant human fibroblast growth factor-2 on bone repair: a dose-escalation prospective trial on patients with osteotomy.

Based on preclinical evidence in animal models, the present study examined the clinical efficacy and safety of recombinant human fibroblast growth factor-2 (rhFGF-2) to accelerate bone repair in a dose-escalation prospective trial. One of three dosages (200, 400 or 800 microg) of rhFGF-2 in a biodegradable gelatin hydrogel was injected during surgery into the osteotomy site of 59 knee osteoarthritis patients undergoing high tibial osteotomy, and 57 of them were monitored for 16 weeks. The rhFGF-2 dose dependently increased the percentage of patients with radiographic bone union, and decreased the average time needed for such union. The percentages of patients with an absence of pain and full-weight bearing were also greater in the higher dosage groups than in the low dosage group, especially in the clinically critical periods 6, 8, and 10 weeks. Neither blood chemistries nor clinical adverse events were associated with the rhFGF-2 dosages. We therefore conclude that the rhFGF-2 in gelatin hydrogel dose dependently accelerated radiographic bone union of a surgical osteotomy with a safety profile at least at the dosages used, suggesting the clinical efficacy of this agent for bone repair.

Adult↗

PubMed2007

Balanced turbo field-echo sequence for MRI of parotid gland diseases.

OBJECTIVE: A balanced turbo field-echo (FE) sequence is a balanced steady-state free precession sequence used for achieving rapid and high imaging. We tested whether this imaging technique is applicable to the diagnosis of parotid gland diseases. CONCLUSION: The balanced turbo FE sequence is a novel alternative MRI technique for the diagnosis of various parotid gland diseases.

Adult↗

PubMed2007

Flexion myelopathy of the thoracic spine. Case report.

The authors report a rare case of surgically treated symptomatic thoracic kyphosis caused by dynamic compression in an elderly man. Myelopathy due to thoracic kyphosis has been reported in patients with congenital kyphosis, Scheuermann dorsal kyphosis, and Cushing disease, but to the authors' knowledge this is the first report of dynamic kyphosis in an elderly person. This otherwise healthy 84-year-old man presented with a 2-year history of progressive difficulty in walking and bilateral leg dysesthesia. Despite several cervical and lumbar surgeries, his symptoms gradually worsened. A radiological examination revealed severe thoracic kyphosis, with a lateral Cobb angle of 59 degrees from T-2 to T-12. On a dynamic computed tomography (CT) myelogram, severe thoracic spinal cord draping and stretching on flexion was demonstrated. On extension, however, imaging studies failed to show draping or stretching. Posterior corrective fusion was performed with instrumentation from T-2 to T-9. Postoperative CT myelography demonstrated no significant spinal cord compression with restoration of the cerebrospinal fluid space anterior to the spinal cord, and the successful correction of the kyphosis to 44 degrees. The patient's neurological sequelae gradually resolved throughout 6 months of follow up.

Aged, 80 and over↗

PubMed2006

Bioactive and bioresorbable cellular cubic-composite scaffolds for use in bone reconstruction.

We used a novel composite fibre-precipitation method to create bioactive and bioresorbable cellular cubic composites containing calcium phosphate (CaP) particles (unsintered and uncalcined hydroxyapatite (u-HA), alpha-tricalcium phosphate, beta-tricalcium phosphate, tetracalcium phosphate, dicalcium phosphate dihydrate, dicalcium phosphate anhydrate or octacalcium phosphate) in a poly-D/L-lactide matrix. The CaP particles occupied greater than or equal to 70 wt% (greater than or equal to 50 vol%) fractions within the composites. The porosities of the cellular cubic composites were greater than or equal to 70% and interconnective pores accounted for greater than or equal to 70% of these values. In vitro changes in the cellular geometries and physical properties of the composites were evaluated over time. The Alamar Blue assay was used to measure osteoblast proliferation, while the alkaline phosphatase assay was used to measure osteoblast differentiation. Cellular cubic C-u-HA70, which contained 70 wt% u-HA particles in a 30 wt% poly-D/L-lactide matrix, showed the greatest three-dimensional cell affinity among the materials tested. This composite had similar compressive strength and cellular geometry to cancellous bone, could be modified intraoperatively (by trimming or heating) and was able to form cortico-cancellous bone-like hybrids. The osteoinductivity of C-u-HA70, independent of biological growth factors, was confirmed by implantation into the back muscles of beagles. Our results demonstrated that C-u-HA70 has the potential as a cell scaffold or temporary hard-tissue substitute for clinical use in bone reconstruction.

Absorbable Implants↗

PubMed2006

Misexpression of Sox9 in mouse limb bud mesenchyme induces polydactyly and rescues hypodactyly mice.

Our previous studies have demonstrated the essential roles of the transcription factor Sox9 in the commitment of mesenchymal cells to a chondrogenic cell lineage and in overt chondrogenesis during limb bud development. However, it remains unknown if Sox9 induces chondrogenesis in mesenchyme ectopically in vivo as a master regulator of chondrogenesis. In this study, we first generated mutant mice in which Sox9 was misexpressed in the limb bud mesenchyme. The mutant mouse embryos exhibited polydactyly in limb buds in association with ectopic expression of Sox5 and Sox6 although markers for the different axes of limb bud development showed a normal pattern of expression. Misexpression of Sox9 stimulated cell proliferation in limb bud mesenchyme, suggesting that Sox9 has a role in recruiting mesenchymal cells to mesenchymal condensation. Second, despite the facts that misexpression of Sonic hedgehog (Shh) induces polydactyly in a number of mutant mice and Shh-null mutants have severely defective cartilage elements in limb buds, misexpression of Sox9 did not restore limb bud phenotypes in Shh-null mutants. Rather, there was no expression of Sox9 in digit I of Hoxa13Hd mutant embryos, and Sox9 partially rescued hypodactyly in Hoxa13Hd mutant embryos. These results provide evidence that Sox9 induces ectopic chondrogenesis in mesenchymal cells and strongly suggest that its expression may be regulated by Hox genes during limb bud development.

Animals↗

PubMed2006

In vitro transformation of mesenchymal stem cells by oncogenic H-rasVal12.

Tissue stem cells may serve as progenitors for malignant tumors derived from the same tissue. Here, we report the establishment of immortalized human mesenchymal stem cells (ihMSC) and tested the feasibility of using ihMSC as presarcomatous cells. Immortalization was achieved by introducing the genes for human telomerase reverse transcriptase and Bmi1. ihMSC retained the potential for multi-directional differentiation of the original MSC. To transform ihMSC, we introduced an oncogenic H-ras(Val12) gene, and established the cell line ihMSC-ras. ihMSC-ras had the phenotype of fully transformed cells and retained adipogenic and chondrogenic, but not osteogenic, potential. Interestingly, ihMSC-ras demonstrated morphological features of autophagy, and inhibition of the ERK pathway suppressed the production of autophagosomes, indicating that ras/ERK signaling is responsible for the induction of autophagy. Thus ihMSC will serve as a material with which to analyze the tumorigenic and differentiation-modifying effects of candidate oncogenes involved in the development of sarcomas.

Cell Culture Techniques↗

PubMed2006

SKL-2841, a dual antagonist of MCP-1 and MIP-1 beta, prevents bleomycin-induced skin sclerosis in mice.

Systemic sclerosis (SSc) is a connective tissue disease characterized by fibrosis and excessive collagen deposition in the skin and various internal organs. In early stages of SSc, the dermis reveals infiltration of inflammatory cells associated with increased collagen synthesis. SKL-2841 was initially synthesized as a novel small molecule antagonist of MCP-1. In this study, we indicated that SKL-2841 also exerts anti-chemotactic activity for MIP-1 beta in mouse spleen cells. In the early stages of bleomycin-induced skin lesions, immunohistochemical analysis showed the expression of both MCP-1 and MIP-1 beta in dermal inflammatory cells. Moreover, intraperitoneal administration of SKL-2841 suppressed the infiltration of inflammatory mononuclear cells and polymorphonuclear cells in the acute phase and also significantly suppressed fibrillization in the chronic phase in bleomycin-induced scleroderma, compared with PBS treatment. These findings suggest that SKL-2841 has potential as a compound for the treatment of conditions associated with skin fibrosis such as SSc.

Animals↗

PubMed2006

Structure and excitation relaxation dynamics of dimethylanthracene dimer in a gamma-cyclodextrin nanocavity in aqueous solution.

Dimethylanthracene (DMA), which exhibits almost no self-association in bulk organic solvents, forms a dimer and emits excimer-like fluorescence in a gamma-cyclodextrin nanocavity in a dilute aqueous solution. The 1Bb and 1La electronic transitions of the DMA dimer split by 2230 and 344 cm(-1), respectively, in a fluorescence excitation spectrum obtained with the excimer-like emission. From these energy splits, the structure of dimer in relation to a dielectric constant inside gamma-CD was discussed on the basis of atom-atom Lennard-Jones potential calculations including Coulombic interactions. Excitation relaxations of DMA in the presence of alpha-, beta-, and gamma-CDs in aqueous solutions were investigated by time-resolved fluorescence. The results suggest that both the hydrated and anhydrated species exist in the alpha- and gamma-CD complexes, while only the anhydrated species exists in the beta-CD complex.

Journal Article↗

PubMed2006

Expression of claudin7 is tightly associated with epithelial structures in synovial sarcomas and regulated by an Ets family transcription factor, ELF3.

Synovial sarcoma, a soft tissue sarcoma that develops in adults, is pathologically subclassified into monophasic spindle synovial sarcoma and biphasic synovial sarcoma with epithelial components. The molecular mechanism building the epithelial components in biphasic synovial sarcoma is totally unknown. Here we investigated claudins, critical molecules in the tight junction, in biphasic synovial sarcoma. Expression profiles of 21 claudins in 17 synovial sarcoma tumor samples, including 9 biphasic tumors, identified claudin4, claudin7, and claudin10 as biphasic tumor-related claudins, and immunohistochemical analyses demonstrated the localization of these claudins in the epithelial component in biphasic tumors, with claudin7 the most closely associated with the epithelial component. The mRNA expression and protein localization of claudin7 coincided with those of the ELF3, an epithelia-specific member of the Ets family of transcription factors. Luciferase reporter assays demonstrated that the presence of the Ets-binding site at -150 in the promoter region of the claudin7 gene was critical for the transcriptional activity, and gel shift and chromatin immunoprecipitation assays confirmed the binding of ELF3 to the Ets site at -150. Inhibition of ELF3 expression by small interfering RNA simultaneously down-regulated the mRNA expression of the claudin7 gene, and the introduction of ELF3 expression in claudin7-negative cell lines induced mRNA expression of the claudin7 gene. Therefore, the induction of claudin7 expression by ELF3 appears critical to the formation of the epithelial structures in biphasic synovial sarcoma.

Base Sequence↗

PubMed2006

Expression profile of amylolytic genes in Aspergillus nidulans.

Aspergillus nidulans possessed 16 putative amylolytic genes consisting of 7 alpha-glucosidase (agdA-F), 7 alpha-amylase (amyA-F), and 2 glucoamylase (glaA and B) genes on the genome. Among them, the agdA, agdB, agdE, agdF, amyA, amyB, amyF, and glaB genes were induced by isomaltose. AmyR, a Zn(II)(2)Cys(6) transcription factor, was required for the induction. The isomaltose-inducible genes possessed at least one consensus sequence for AmyR binding, 5'-CGGN(8)CGG, on each promoter region. None of the amylolytic genes was induced by maltose. The mRNA levels of the amylolytic genes except for agdC, amyD, and amyG increased under carbon-starved conditions. Release from CreA-dependent carbon catabolite repression was the main cause of the increase, but, the mRNA levels of agdB, agdF, amyB, amyF, and glaB increased to some extent even in a creA mutant. Therefore, both CreA-dependent and -independent mechanisms are involved in the up-regulation of the amylolytic genes under carbon-starved conditions.

Amylases↗

PubMed2006

Pore throat size and connectivity determine bone and tissue ingrowth into porous implants: three-dimensional micro-CT based structural analyses of porous bioactive titanium implants.

A porous structure comprises pores and pore throats with a complex three-dimensional (3D) network structure, and many investigators have described the relationship between average pore size and the amount of bone ingrowth. However, the influence of network structure or pore throats for tissue ingrowth has rarely been discussed. Four types of bioactive porous titanium implants with different pore sizes and porosities (6mm in diameter and 15 mm long) were analyzed using specific algorithms for 3D analysis of interconnectivity based on a micro focus X-ray computed tomography system. In vivo histomorphometric analysis was performed using the very same implants implanted into the femoral condyles of male rabbits for 6 and 12 weeks. This matching study revealed that more poorly differentiated pores tended to have narrow pore throats, especially in their shorter routes to the outside. In addition, for assessment of the entire implant, we proposed new two indices that represent the degree of bone and tissue ingrowth into an implant by considering the effect of narrow pore throats. Data obtained suggest that this sort of novel analysis is useful for evaluating bone and tissue ingrowth into porous biomaterials.

Algorithms↗

PubMed2006

Successful storage of peripheral nerves using University of Wisconsin solution with polyphenol.

We have previously reported that green tea polyphenol can preserve peripheral nerve segments for up to 1 month. In this study, we investigated the effect on peripheral nerve preservation of adding polyphenol to the conventional University of Wisconsin solution (UW solution), which has been widely used for organ storage. Twenty millimeter-long sciatic nerve segments, harvested from Lewis rats, were immersed in UW solution containing polyphenol (1 mg/mL) for 1 week and then in UW solution alone at 4 degrees C for 3 additional weeks before transplantation into recipient Lewis rats. Neural cell viability of the preserved nerve segments was confirmed by vital staining (calcein-AM/ethidium homodimer), electron microscopy, and genomic studies. Morphologically, nerve regeneration was similar to that of fresh isografts and superior to that of grafts stored with UW solution alone. Moreover, the electrophysiological results were equal to those of fresh isografts. Polyphenol has the potential to be used for peripheral nerve storage and could be useful for routine peripheral nerve banking.

Actins↗

PubMed2006

Development of ulcerative colitis during the course of rheumatoid arthritis: Association with selective IgA deficiency.

A 56-year-old woman with a 29-year history of rheumatoid arthritis (RA) was admitted to the hospital, complaining of high fever, abdominal pain and severe bloody diarrhea. Colonoscopy revealed friable and edematous mucosa with spontaneous bleeding, diffuse erosions and ulcers extending from the rectum to the distal transverse colon. Histopathological findings of rectal biopsies were compatible with ulcerative colitis (UC). Being diagnosed as having severe active left-side UC, she was successfully treated with intravenous methylprednisolone followed by prednisolone and leukocytapheresis. Laboratory tests revealed low serum and saliva IgA levels, which might play a role in the development of UC. To our knowledge, this is the first case of UC occurring during the course of RA, accompanied by selective IgA deficiency.

Anti-Inflammatory Agents↗

PubMed2006

Preparation and evaluation of a chiral stationary phase covalently bound with a chiral pseudo-18-crown-6 ether having a phenolic hydroxy group for enantiomer separation of amino compounds.

In order to develop a chiral stationary phase (CSP), which has even higher separation ability than the corresponding commercially available crown ether based CSP (OA-8000 having a pseudo-18-crown-6 ether with an OMe group as a selector), chemically bonded type CSP having a phenolic OH group on a crown ring was developed. Normal mobile phases with or without acid additive can be used with this OH type CSP in contrast to the conventional OMe type CSP which has a neutral chiral selector. Enantiomers of 25 out of 27 amino compounds, including 20 amino acids, 5 amino alcohols, and 2 lipophilic amines, were efficiently separated on a column with this CSP. Nine amino compounds out of 27 were separated with better separation factors than the corresponding OMe type CSP. It is noteworthy that the chromatography on this CSP exhibited excellent enantiomer-separations for amines and amino alcohols when triethyl amine was used as an additive in the mobile phase. Comparison of enantiomer separation ability on this OH type of CSP and on the OMe type of CSP and correlation between the enantioselectivity in chiral chromatography and that of the corresponding model compounds in solution imply that the chiral separation arose from chiral recognition in host guest interactions.

Amines↗

PubMed2006

Isoprene emission from tropical trees in Okinawa Island, Japan.

This study surveyed isoprene emission from 42 indigenous and exotic tropical trees in subtropic Okinawa, Japan. Of the 42 trees studied, 4 emitted isoprene at a rate in excess of 20 microg g(-1)h(-1), and 28 showed the rates of 1-10 microg g(-1)h(-1). The remainder emitted less than 1 microg g(-1)h(-1). The majority of trees in this study may therefore fall within the lower emitting species. However, species in Moraceae that is indigenous in Okinawa emitted isoprene at relatively higher rates with an average of 14.2 microg g(-1)h(-1). The highest emission rate of 107.1 microg g(-1)h(-1) for Ficus virgata yielded the area basis rate of 47.4 nmol m(-2)s(-1), which is almost equivalent to the rate of high emitting species. Furthermore, a linear relationship between light intensity and isoprene emission was noted with Ficus virgata up to 1700 micromol m(-2)s(-1). These findings may show the potential importance of subtropical areas as sources of isoprene to the atmosphere.

Butadienes↗

PubMed2006

Comparison of the anatomical risk for vertebral artery injury associated with the C2-pedicle screw and atlantoaxial transarticular screw.

STUDY DESIGN: We evaluated the trajectories of atlantoaxial transarticular and C2-pedicle screws in 3 dimensions using computerized tomography. OBJECTIVE: To compare the anatomic risk for vertebral artery injury associated with C2-pedicle and atlantoaxial transarticular screws. SUMMARY OF BACKGROUND DATA: The atlantoaxial fixation technique using C1-lateral mass screws combined with C2-pedicle screws is considered a safer technique for preventing vertebral artery injury than atlantoaxial transarticular fixation. However, few reports have compared the anatomic risk of vertebral artery injury associated with C2-pedicle screws with that of transarticular screws. METHODS: A total of 62 consecutive patients with cervical lesions were evaluated using 3-dimensional images reconstructed by a computer-assisted navigation system. We compared the maximum possible diameters of the atlantoaxial transarticular screw and C2-pedicle screw trajectories, and examined whether the maximum possible diameters were limited by the height or width of the bony structure in screw trajectories < or = 4 mm in diameter. RESULTS: Mean maximum possible diameters did not differ significantly between the trajectories of 124 atlantoaxial transarticular and 124 C2-pedicle screws. In screw trajectories < or = 4 mm in diameter, 57.1% of transarticular screw trajectories were limited by the height of the bony structure, and all pedicle screw trajectories were limited by the width. CONCLUSIONS: C2-pedicle screw placement has nearly the same anatomic risk of vertebral artery injury as transarticular screw placement. Preoperative 3-dimensional evaluation may be useful for choosing the best surgical technique.

Adult↗