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Biomedical subjects

A Uchida

Publications and source records attributed to A Uchida.

At least 91 records · Page 5Linked to original sources

TRAIL causes cleavage of bid by caspase-8 and loss of mitochondrial membrane potential resulting in apoptosis in BJAB cells.

A new member of the TNF family, tumor necrosis factor-related apoptosis-inducing ligand (TRAIL), has been shown to induce apoptosis. However, the mechanism for TRAIL-induced apoptosis remains to be clarified. SDS-PAGE and Western blot analysis showed that cleavage of Bid was induced by a 1-h incubation of BJAB cells with TRAIL and was blocked by a caspase-8 inhibitor. Flow cytometry demonstrated that loss of mitochondrial membrane potential in BJAB cells began about 1.5 h after the treatment with TRAIL and was apparent at 2 h in comparison with the control. DNA ladder formation, which is characteristic for apoptosis, in the cells treated with TRAIL was detected at 2 h and observed most effectively at 3 h. The time course study suggests that TRAIL causes cleavage of Bid via activation of caspase-8, subsequently the loss of mitochondrial membrane potential, resulting in apoptosis in BJAB cells.

Apoptosis↗

Expression of gp145trkB in the early stage of Schwann cell tube formation.

We investigated the expression of tyrosine kinase receptors (trkBs) and their ligand, brain-derived neurotrophic factor (BDNF), in isolated sciatic nerve segments of rat in a silicone chamber model devoid of axonal contact, using an immunohistochemical technique. The receptor gp 95trkB was not detected during the first 3 weeks but was strongly expressed in the nerve stumps on day 21. In contrast, gp 145trkB immunoreactivity was readily detected in the Schwann cells in the nerve stumps between days 1 and 7 after isolation. Immunohistochemical analysis using anti-S-100 antibody demonstrated that Schwann cell tube formation within the silicone chamber had been completed by the third week, which suggested that gp 95trkB began to be produced by Schwann cells after completion of Schwann cell tube formation. Considering the low level of BDNF production during the first week, molecules other than BDNF may be ligands for gp 145trkB in the silicone chamber model devoid of axonal contact.

Animals↗

Purification and characterization of an intracellular heat-stable proteinase (pernilase) from the marine hyperthermophilic archaeon Aeropyrum pernix K1.

A novel intracellular serine proteinase from the marine aerobic hyperthermophilic archaeon Aeropyrum pernix K1 (JCM 9820) that we designated pernilase was purified by ammonium sulfate precipitation, anionic-exchange chromatography, affinity chromatography, and gel filtration chromatography. The purified enzyme was composed of a single polypeptide chain with a molecular mass of 50 kDa as determined by SDS-PAGE. The proteinase had a broad pH profile (pH 5-10) with an optimum pH of 9.0 for peptide hydrolysis. The optimum temperature for enzyme activity was 90 degrees C. The enzyme was strongly inhibited by diisopropyl fluorophosphate (DFP) and phenylmethyl sulfonylfluoride (PMSF), suggesting that it corresponds to a serine proteinase. The enzyme was highly resistant to the reducing agents dithiothreitol and 2-mercaptoethanol but sensitive to the denaturing reagents guanidine-HCl and urea and also to the detergent sodium dodecyl sulfate (SDS). Pernilase showed high substrate specificity for Boc-Leu-Gly-Arg-MCA peptide. Thermostability of this enzyme showed half-lives of 85min at 100 degrees C and 12 min at 110 degrees C.

Amino Acid Sequence↗

Biological performance of a three-dimensional fabric as artificial cartilage in the repair of large osteochondral defects in rabbit.

A new artificial cartilage [A three-dimensional fabric (3-DF) comprising ultra-high molecular weight polyethylene fiber with a triaxial three-dimensional structure and coated with hydroxyapatite] was used to repair large osteochondral defects in rabbit knees. The knees with the 3-DF implanted in the osteochondral defects were evaluated 2-24 weeks after the operation macroscopically and microscopically. The 3-DF coated with hydroxyapatite was fixed firmly to the subchondral bone with the newly formed bone into and around the 3-DF. Hyaline-like cartilage was formed to a certain extent on the surface of the 3-DF, and the surface damage of the patella opposed to the 3-DF was minimal. In addition, the 3-DF caused no adverse effects such as particle disease, infection or severe synovitis during the experimental period. Consequently, the 3-DF seems to have successful biocompatibility in this rabbit experiment.

Animals↗

Gene expression during osteoclast-like cell formation induced by antifusion regulatory protein-1/CD98/4F2 monoclonal antibodies (MAbs): c-src is selectively induced by anti-FRP-1 MAb.

Human blood monocytes can differentiate into osteoclast-like cells when they are cultured in the presence of anti-FRP-1. Messenger (mRNA) expression of markers related to osteoclasts was analyzed during differentiation of osteoclasts from monocytes. As markers related to osteoclasts, we selected cathepsin-K, carbonic anhydrase (CA) II, vacuolar H(+)-ATPase (v-ATPase), vitronectin receptor (VNR), tartrate-resistant acid phosphatase (TRAP), osteopontin (OPN), galectin-3, c-src, c-fos, and c-fms. The mRNAs other than c-src mRNA were expressed in freshly isolated monocytes or monocytes incubated with control antibody or anti-FRP-1 monoclonal antibody (MAb) for 14 days. Of these mRNAs, cathepsin-K, CA II, v-ATPase, VNR, TRAP, OPN, and c-fms mRNAs were expressed at higher levels in the osteoclast-like cells than those in monocytes cultured with control antibody. On the other hand, galectin-3 mRNA was expressed at lower levels in the osteoclast-like cells, and there was no significant difference in c-fos mRNA expression between the monocytes cultured with control antibody and anti-FRP-1 MAb. c-src mRNA could not be detected in monocytes freshly isolated or incubated with control antibody. Surprisingly, expression of c-src mRNA was induced in monocytes by anti-FRP-1 MAb and was detectable as early as 3 h after anti-FRP-1 MAb treatment, indicating that c-src is selectively induced by anti-FRP-1 MAb treatment. Furthermore, the osteoclast-like cells expressed calcitonin receptor. Receptor activator of NF-kappaB (RANK) mRNA was detectable in freshly isolated monocytes or monocytes cultured with control antibody or anti-FRP-1 MAbs. Maximal expression of RANK was observed in osteoclast-like cells. On the other hand, no receptor activator of NF-KB ligand (RANKL) mRNA was detectable in any of the samples, suggesting that anti-FRP-1 mAb can induce osteoclast-like cells from blood monocytes without RANKL.

Antibodies, Monoclonal↗

Schwann cells can induce collateral sprouting from intact axons: experimental study of end-to-side neurorrhaphy using a Y-chamber model.

Axonal regeneration across end-to-side neurorrhaphy has recently been reported; however, neither the mechanism by which collateral sprouting from intact axons is elicited, nor the origin of the regenerating axons are known. There has even been controversy over the presence of collateral axonal sprouting from intact axons altogether. This reported experimental study was designed to clarify these questions. A rat sciatic nerve model was used. To avoid any mechanical damage to the donor nerve during the procedure, a Y-shaped silicone chamber was employed instead of direct suture. Axonal regeneration from the intact tibial nerve across the gap into the peroneal nerve was assessed using a retrograde neurotracer and immunohistochemical staining. Axonal regeneration across the gap was observed in 66 percent of the animals. The neurotracer evaluation clearly showed that all regenerating axons were sensory axons from the dorsal root ganglia. The authors concluded that Schwann cells from the distal wallerian degeneration of nerve segments did elicit collateral axonal sprouting from intact sensory axons, but not from motor axons in end-to-side neurorrhaphy. Invasion of the Schwann cells into the epineurial layer was the crucial step for the initiation of collateral axonal sprouting from the intact axons.

Animals↗

Mandibular-position sensation during sedation by administration of nitrous-oxide (N2O) gas.

Our aim was to confirm the influence of N2O gas on mandibular-position sensation. The subjects in this study were eight healthy adults. Each subject was asked to hold the reference stick for five seconds between the central tooth in his or her upper and lower jaws. Then, the reference stick was replaced by the test sticks with different thickness, each of which the subjects were again asked to hold at the same position for five seconds. The subjects were instructed to determine, based on judgement of the interincisal distance, whether the thickness of each test stick was larger or smaller than the reference stick. A series of trials was administered to each subject using all eight sizes of test sticks. We compared the ability of the subjects to discriminate mandibular position both before and after the application of a vibrating stimulus to the masticatory muscles, and before and during the administration of N2O gas. Discrimination ability was significantly decreased after the application of the vibrating stimulus. However, during the administration of N2O gas, no significant difference in discrimination ability was observed between before and after the vibrating stimulus. The results of this study indicated that N2O gas had an inhibitory effect on gamma-motor neuron activity, which is presumed to be mediated to some extent through the higher central nervous system. This is the case because the gamma-motor neurons are generally activated by vibrating stimuli applied to the muscle causing decreased discrimination ability of mandibular position. Thus, we conclude that mandibular-position sensation was influenced by N2O gas during the administration of N2O gas.

Adult↗

Gene therapy of chondrosarcoma using retrovirus vectors encoding the herpes simplex virus thymidine kinase gene.

Human chondrosarcoma cells (HCS-TG) were transduced with the gene for a herpes simplex virus thymidine kinase (HSV-tk) or Escherichia coli beta-galactosidase (lacZ). We investigated the cytotoxicity of human chondrosarcoma bearing an HSV-tk gene after treatment with ganciclovir. Chondrosarcoma cells bearing an HSV-tk gene were more sensitive than non-transduced cells. Coculturing with chondrosarcoma cells bearing both an HSV-tk gene (HCS-TG-tk) and lacZ gene (HCS-TG-Z) in various ratios showed a bystander effect. Chondrosarcoma implanted in nude mice were injected with HCS-TG-tk cells. After 4 weeks, the growth of tumors was significantly prevented.

Animals↗

The zymogen prothrombin stimulates cell locomotion and calcium influx in murine osteosarcoma cells by different mechanism from thrombin.

The coagulation system plays a role in tumor invasion and metastasis. Prothrombin was previously found to stimulate the motility of melanoma cells. We compared the effect of prothrombin and thrombin on cell motility, calcium influx and actin stress filament contraction in highly metastatic osteosarcoma LM8 cells. Prothrombin and thrombin stimulated chemotaxis and chemokinesis of LM8 cells in a dose-dependent manner. Calcium influx was significantly increased after stimulation with thrombin and prothrombin. Thrombin and prothrombin markedly stimulated actin contraction in LM8 cells. Hirudin inhibited the effects of thrombin but not those of prothrombin. Prothrombin appears to stimulate cell locomotion, actin contraction, calcium influx in LM8 cells by different mechanism from thrombin.

Actins↗

Relation between hemoglobin discordance and adverse outcome in monochorionic twins.

BACKGROUND: The effects of discrepancies in the birth weight and hemoglobin concentration ([Hb]) at the birth on infant outcome have not been fully investigated in monochorionic (MC) and dichorionic (DC) twins. METHODS: A retrospective study of 95 twin pregnancies. All 190 neonates (68 MC twins and 122 DC twins) were admitted to the neonatal intensive care unit, and the [Hb] was determined soon after birth. The [Hb] discordance represents the [Hb] of the smaller twin expressed as a percentage of the [Hb] of the larger twin. Infants were followed-up until 1 year of corrected age and their psychomotor development was evaluated between 12 and 18 months. RESULTS: One DC infant died by 1 year of age. Twelve MC infants and eight DC infants had disabilities by 1.5 year of age. In the MC group, 11 infants (28%) of 20 pairs with an intertwin [Hb] discordance >100 had disabilities compared with one (3.6%) of 14 pairs with an [Hb] discordance < or = 100 (p<0.05). A similar trend was observed in the DC group, but the difference was not significant. The incidence of disabled infants increased as the intertwin birth weight discordance increased in MC twins. CONCLUSIONS: An adverse outcome at 1 year of age in MC twins was associated with a greater birth weight discordance and an [Hb] disordance that indicated polycythemia in the smaller twin.

Adult↗

Restoring normal gait after limb salvage procedures in malignant bone tumours of the knee.

Eleven patients exhibiting decreased strength of knee extension following wide resection and prosthetic reconstruction for malignant bone tumors of the knee performed gait exercises with compensatory muscle training. Two patients whose knee extension strength was assessed as manual muscle test (MMT) grade 4 were able to develop a gait with double knee action and to maneuver stairs, step-by-step, due to compensation by the gluteus maximus, biceps femoris, and gastrocnemius muscles. Four patients whose knee extension strength was less than MMT grade 4, and whose ankle dorsal and plantar flexion was MMT grade 4 or higher, acquired the ability to go up and down stairs step-by-step, although their gait pattern was a knee-extended gait. Electromyographic studies demonstrated continuous discharges of the gluteus maximus, biceps femoris, and gastrocnemius muscles during the stance phase as compensation for decreased strength in knee extension.

Adolescent↗

[Recent advances in management of musculoskeletal tumors].

Neoadjuvant chemotherapy plays a significant role in improving the prognosis of malignant bone tumors such as osteosarcoma and Ewing's sarcoma. Doxorubicin, cisplatin and high-dose methotrexate are key drugs in the chemotherapy for osteosarcoma. Intraarterial administration of the drugs is very effective in local control, so that wide resection of the tumor may be performed safely. The efficacy of preoperative chemotherapy is evaluated by the histological necrosis rate of the resected tumor after which postoperative drugs will be chosen. Limb salvage is becoming more and more common, and an excellent limb can be reconstructed using an artificial joint. In soft tissue sarcomas, brachytherapy can make the surgical resection margin narrower, resulting in good function of the extremities. Fragility of bone caused by metastatic bone diseases is reversed by surgical intervention.

Antineoplastic Combined Chemotherapy Protocols↗

Effects of placental chorionicity on outcome in twin pregnancies. A cohort study.

OBJECTIVE: To examine the effects of the chorionicity of the placenta on infant outcome at 1 year of age in twin pregnancies. STUDY DESIGN: Cohort study and retrospective review of the medical records of 44 monochorionic (MC) and 164 dichorionic (DC) twin gestations that had been followed at our institution since < 20 weeks' gestation. Physical and neurologic status was assessed at 1 year of corrected age in infants born to these 208 women. RESULTS: Adverse infant outcomes, such as death, cerebral palsy and mental retardation, occurred in 9 (10%) of 88 MC infants (4 deaths and 5 disabled infants) as compared with 12 (3.7%) of 328 DC infants (6 deaths and 6 disabled infants) (P < .05). Although delivery occurred one week earlier in MC than in DC twins (34.7 +/- 2.8 vs. 35.7 +/- 2.3 weeks, P < .01), there was no significant difference in gestational age at birth or birth weight between the 9 MC and 12 DC infants with adverse outcomes. A presumptive antenatal diagnosis of twin-twin transfusion syndrome (TTTS) was made in 14 (32%) of the 44 MC twin gestations. TTTS was considered to be responsible for adverse outcome in 7 MC infants. All 9 MC infants with adverse outcomes and 4 (33%) of 12 DC infants with adverse outcomes belonged to pairs that had weight discordance > or = 25% (P < .01). CONCLUSION: MC twins had an increased risk of adverse outcomes as compared with DC twins, mainly because of TTTS. In both MC and DC twins, a birth weight discordance > or = 25% was associated with adverse infant outcomes. The number of infants with disabilities at 1 year of age was equal to the number of deaths.

Adult↗

Intraoperative extracorporeal autogenous irradiated bone grafts in tumor surgery.

There are several procedures for reconstruction of bony defects after resection of malignant musculoskeletal tumors. The clinical results of intraoperative extracorporeal autogenous irradiated bone grafts in 20 patients with musculoskeletal tumors are discussed. The authors' method of treatment consists of: (1) wide en bloc resection of the tumor with involved bone; (2) curettage of the tumor from the resected bone; (3) extracorporeal irradiation with 50 Gy as a bolus single dose to the isolated bone; and (4) reimplantation of the irradiated bone into the host with fixation devices. Twelve bone sarcomas and eight soft tissue sarcomas with bone involvement were treated surgically with this reconstruction method after wide resection of the tumors. The irradiated bone was used as an intercalary graft in seven cases, as an osteoarticular graft in 11 cases, and as a hemicortical graft in two cases. The theoretical advantages of this method are certain sterilization of tumor cells with radiation, easy availability and good adaptation of size and shape, no risk of disease transmission, preservation of bone stock and ligamentous tissue, and no immunologic reaction. Radiologically, bony union occurred in 23 of 29 (79%) osteotomy sites. The overall radiographic evaluation rating was 74% and the functional rating was 73% according to the International Society of Limb Salvage rating system. Nonunion (20%) and infection (15%) were the two major complications. Preservation of the tendon insertions and ligamentous structures of the irradiated bone seemed to restore excellent joint function. No local recurrence was detected from the irradiated bones during the mean followup of 45 months. These results indicate intraoperative extracorporeal irradiated bone graft can be a widely applicable method for reconstruction in tumor surgery.

Adolescent↗

Splicing patterns of type XI collagen transcripts act as molecular markers for osteochondrogenic tumors.

Primary transcripts for three distinct alpha chains of the type XI collagen molecule (alpha1(XI), alpha2(XI) and alpha3[XI]) undergo tissue-specific alternative splicing during the process of osteochondrogenesis. In the present study, we analyzed the splicing patterns of type XI collagen genes in osteochondrogenic tumors as well as in various normal tissues using the reverse transcription-polymerase chain reaction method. Analysis of normal subjects revealed the coordinated expression of short alpha1(XI), alpha2(XI) and alpha3(XI) transcripts in the normal differentiated cartilage. Osteochondroma followed this pattern, reflecting the highly chondrogenic phenotype of this benign tumor. Another benign tumor, chondroblastoma, exclusively expressed the long alpha1(XI) transcript, probably reflecting the lack of a chondrogenic nature. Among malignant chondrogenic tumors, the splicing patterns of type XI collagen transcripts were more complex, showing dissociated expression of long alpha1(XI) and short alpha2(XI) mRNAs. This expression pattern may reflect heterogeneous cell populations and may also reflect various levels of cell differentiation in malignant tumors. In addition, short alpha3(XI) expression switched to the long transcript as chondrosarcomas became more aggressive. Thus, the alternative splicing of type XI collagen genes seems to be oncodevelopmentally regulated and splicing analysis may therefore be a useful marker for chondrogenic tumors.

Alternative Splicing↗

New ferromagnetic bone cement for local hyperthermia.

We have developed a ferromagnetic bone cement as a thermoseed to generate heat by hysteresis loss under an alternate magnetic field. This material resembles bioactive bone cement in composition, with a portion of the bioactive glass ceramic component replaced by magnetite (Fe3O4) powder. The temperature of this thermoseed rises in proportion to the weight ratio of magnetite powder, the volume of the thermoseed, and the intensity of the magnetic field. The heat-generating ability of this thermoseed implanted into rabbit and human cadaver tibiae was investigated by applying a magnetic field with a maximum of 300 Oe and 100 kHz. In this system, it is very easy to increase the temperature of the thermoseed in bone beyond 50 degrees C by adjusting the above-mentioned control factors. When the temperature of the thermoseed in rabbit tibiae was maintained at 50 to 60 degrees C, the temperature at the interface between the bone and muscle (cortical surface) surrounding the material rose to 43 to 45 degrees C; but at a 10-mm distance from the thermoseed in the medullary canal, the temperature did not exceed 40 degrees C. These results demonstrate that ferromagnetic bone cement may be applicable for the hyperthermic treatment of bone tumors.

Animals↗

Variation in tooth morphology of Gorilla gorilla.

Gorilla gorilla exemplifies a species that shows considerable variation in habitat, behaviour, genetic structure and morphology. This study examines variation of dental morphology in gorillas. Despite the marked size dimorphism, there are no significant shape differences between the sexes within subspecies. Differences in dental morphology, including tooth cusp proportions between the western G. g. gorilla and the eastern G. g. beringei are considerable. Although more similar to G. g. beringei than to the western G. g. gorilla, G. g. graueri also shows distinct morphological features. This indicates that the morphology of G. g. graueri is not merely intermediate, and genetic isolation between the two eastern subspecies could have had a substantial influence. Such extensive variation in dental morphology in Gorilla gorilla can be considered to be the result of an interesting combination of factors, including local dietary adaptations.

Animals↗

Variation in tooth morphology of Pongo pygmaeus.

This study examines the postcanine morphology of Pongo pygmaeus, identifying sexual, populational, and subspecific differences, with particular focus on the cusp area proportions. Little differences between the sexes were found in molar shapes, and cusp area proportions can be considered population specific. Inter-population differences are significant between the two Bornean populations, with the Kapuas River as a biogeographic barrier, and are as great as inter-subspecies (Borneo-Sumatra) differences.

Animals↗