Biomedical subjects

M Oka

Publications and source records attributed to M Oka.

At least 361 records · Page 20Linked to original sources

PubMed1995

Autogenous callo-osseous grafts for the repair of osteochondral defects.

We describe a new method of biological repair of osteochondral defects. In rabbit knees an osteochondral defect was reconstructed with a callo-osseous graft made of a superficial sheet of medullary fracture callus attached to a base of cancellous bone. This was taken from the iliac bone of the same animal which had been osteotomised ten days earlier. The reparative tissues were evaluated for 24 weeks by quantitative histology, biochemical analysis of the uronic acid content, and immunohistochemical staining of collagen constituents. The callo-osseous graft provided significantly faster and better repair of the articular surface than an untreated defect or a callo-osseous graft in which the cells had been devitalised by irradiation before transplantation. Our findings indicate that the callo-osseous graft contributes to the repair process by providing both favourable extracellular matrices and pluripotential mesenchymal cells. Our study tested the hypothesis that early medullary callus generates hyaline cartilage instead of bone after transfer to an articular surface.

Adaptation, Physiological↗

PubMed1995

[Measurement of anti-acetylcholine receptor antibody using human rhabdomyosarcoma cell line].

We developed a radioimmunoassay for AChR Ab using AChR solubilized from membrane extracts of human rhabdomyosarcoma cell line (RD cell). The binding affinity of this AChR to the alpha-Bungarotoxin (alpha-BTX) was similar to that of AChR from cell surface. Lyophilized AChR was stable within two months at -20 degrees C. In our assay system, sera from patients with Myasthenia Gravis (MG) showed markedly high titers and frequency (94%) of anti-AChR Ab. On the other hand, almost all sera from normal and non-myasthenic patients such as rheumatoid arthritis and thyroid disease were negative (< 0.2nmol/l). Our assay using AChR from RD cells shows sufficient results in the intra- and inter-CV values, and recovery and dilution tests. AChR Ab titers of our assay showed good correlations with those measured by AChRs from both human ischemic muscles and TE671 cells (commercial kit). The radioimmunoassay for AChR Ab using the membrane extracts of RD cells has high sensitivity, specificity and stability. Therefore, the assay is valuable as the routine assay for the diagnosis of MG.

Autoantibodies↗

PubMed1995

Small depressed cancer of the large bowel: report of three cases.

This report describes three patients in whom colonoscopy detected small depressed cancers (without an elevated component) that had invaded the submucosa. They represent 0.4% (3/884) of all patients with invasive cancers and 3% (3/101) of patients with submucosal cancers in the National Cancer Center Hospital between January 1990 and February 1994. This type of cancer may have been overlooked in the past because of its small, flat nature. A slight deformity of the lumen, a faint color change (slightly reddish), and a loss of a vascular network pattern were important colonoscopic findings. Small depressed cancers may follow a different pathway to advanced cancer than polypoid cancers, although both pathways are included in the adenoma-carcinoma sequence. We should be aware of these lesions in our efforts to detect colorectal cancers in the early stage.

Adenocarcinoma↗

PubMed1995

Posttranscriptional regulation of the c-myb proto-oncogene in estrogen receptor-positive breast cancer cells.

We have determined that expression of the c-myb proto-oncogene is associated with estrogen receptor (ER) status and not with tumor progression in human breast epithelial cells. Analysis of normal, immortalized, nontumorigenic, and tumorigenic mammary epithelial cells showed that only ER+ tumor cell lines expressed readily detectable levels of c-myb mRNA and a Mr 75,000 protein that was the same size as the c-myb transcripts and protein products present in hematopoietic cells. In this report we show that c-myb mRNA and protein levels are down-regulated during estrogen withdrawal. A 20-fold increase in c-myb mRNA and protein expression was observed upon addition of beta-estradiol to the culture medium. Nuclear run-on transcription analyses showed that c-myb was transcribed at the same rate in the presence and absence of estrogen, suggesting that c-myb mRNA accumulation was regulated at a posttranscriptional level. To provide additional evidence that c-myb mRNA was dependent on ER expression, we examined c-myb mRNA levels in MCF-7 cells selected for resistance to antineoplastic drugs. c-myb expression was decreased only in cell lines that showed concomitant loss of ER expression. Moreover, c-myb mRNA was expressed and modulated by estrogen in ER-, MDA-MB-231 cells stably transfected with a human ER gene. When considered together, these data indicate that c-myb mRNA levels are regulated by estrogens and further suggest that this proto-oncogene plays a role in the biology of ER+ breast tumor cells.

Breast↗

PubMed1994

Muscarinic receptor-mediated calcium efflux from cultured bovine adrenal chromaffin cells.

The effect of stimulation of the muscarinic receptor on Ca2+ mobilization in cultured bovine adrenal chromaffin cells was examined. Acetylcholine (ACh) increased the uptake of 45Ca2+ and [Ca2+]i whose levels decreased with time after reaching peaks. It also enhanced the efflux of 45Ca2+ from the cells. Its effect was inhibited by the specific muscarinic receptor antagonist atropine (Atr), but not by the nicotinic receptor antagonist hexamethonium (C6). The increase in muscarine (Mus)-stimulated 45Ca2+ efflux was reduced concentration-dependently by deprivation of extracellular Na+. These results suggest that muscarinic stimulation of the ACh receptor stimulates Na+/Ca2+ exchange in cultured bovine adrenal chromaffin cells.

Acetylcholine↗

PubMed1994

Mechanism of histamine-induced calcium efflux from cultured bovine adrenal chromaffin cells: possible involvement of an Na+/Ca2+ exchange mechanism.

The effect of stimulation of the histamine receptor on Ca2+ mobilization in cultured bovine adrenal chromaffin cells was examined. Histamine (10(-5) M) increased the intracellular free Ca2+ ([Ca2+]i) to a peak in the presence or absence of extracellular Ca2+, followed by decrease with time. Histamine (10(-8)-10(-5) M) also stimulated 45Ca2+ efflux from cultured bovine adrenal chromaffin cells in a concentration dependent manner. Its stimulatory effect on 45Ca2+ efflux was inhibited by the specific histamine H1 receptor antagonist mepyramine. The increase in histamine-stimulated 45Ca2+ efflux was inhibited by deprivation of extracellular Na+ and by the Na+/Ca2+ exchange inhibitor amiloride. In addition, histamine stimulated 22Na+ influx into the cells, and this action was inhibited by amiloride. These results suggest that stimulation of the histamine H1 receptor regulates Na+/Ca2+ exchange in cultured bovine adrenal chromaffin cells.

Adrenal Medulla↗

PubMed1994

Use of intra-operative ultrasonography to detect a small foreign body in the soft tissue of the upper lip.

Real-time ultrasonography was used to detect a small foreign body (a broken tip of a barbed broach) which had migrated into the upper lip of a 26-year-old female from the bone defect of the upper canine apex during root canal treatment 10 months previously. Intra-operative use of ultrasound was of great-help in clarifying the positional relationship between the foreign body and the surgical instruments used, and lead to a satisfactory result.

Adult↗

PubMed1994

Inhibition by ouabain of palytoxin-induced catecholamine secretion and calcium influx into cultured bovine adrenal chromaffin cells.

The effect of ouabain on palytoxin (PTX)-induced catecholamine secretion from cultured bovine adrenal chromaffin cells was examined in relation to its effect on calcium (Ca2+) influx into the cells. Ouabain showed concentration-dependent inhibition of catecholamine secretion induced by PTX. Ouabain also inhibited [45Ca]2+ influx induced by PTX, this inhibition being parallel with that of catecholamine secretion. The inhibitory effects of ouabain on PTX-induced catecholamine secretion and [45Ca]2+ influx were both overcome by increasing the concentrations of PTX, indicating that ouabain inhibited the actions of PTX in a competitive manner. These results suggest that the ouabain-sensitive (or-binding) site on the cell membrane might be the target site of action of PTX, which causes an increase in Ca2+ permeability and initiation of catecholamine secretion.

Acrylamides↗

PubMed1994

Tertiary structure of erabutoxin b in aqueous solution as elucidated by two-dimensional nuclear magnetic resonance.

The three-dimensional structure of erabutoxin b, a short-chain neurotoxic peptide purified from the venom of the sea snake Laticauda semifasciata, was determined in aqueous solution by two-dimensional proton nuclear magnetic resonance and simulated annealing-based calculations. On the basis of 883 assigned nuclear Overhauser effect (NOE) connectivities, 676 final distance constraints were derived and used together with 38 torsion angle (phi, chi 1) constraints, four distance constraints derived from disulfide bridges and 30 distance constraints derived from hydrogen bonds. A total of 14 converged structures were obtained from 50 runs of calculations. The atomic root-mean-square difference about the mean coordinate positions (excluding the residues 18 to 22) is 0.60 A for backbone atoms (N, C alpha and C'). The protein consists of a core region from which three finger-like loops emerge outwards. It includes a short, two-stranded antiparallel beta-sheet of residues 2 to 5 and 13 to 16, a three-stranded antiparallel beta-sheet involving residues 23 to 30, 35 to 41 and 50 to 56, and four disulfide bridges in the core region. Comparison with two crystal structures of erabutoxin b at 1.4 A and 1.7 A resolution indicated that the solution and the crystal structures were very similar, but less defined regions were observed at the localized region of the tip of the central loop and the outside of the third loop in solution. Other short-chain alpha-neurotoxins showed structural characteristics similar to those of erabutoxin b.

Amino Acid Sequence↗

PubMed1994

Effects of the potassium channel openers cromakalim and pinacidil on catecholamine secretion and calcium mobilization in cultured bovine adrenal chromaffin cells.

The effects of two K+ channel openers, cromakalim and pinacidil, on voltage-dependent and receptor-mediated catecholamine secretion and Ca2+ mobilization in bovine adrenal chromaffin cells were studied to determine the role of membrane K+ channels in the regulation of a Ca(2+)-dependent secretory process. Both cromakalim and pinacidil stimulated the efflux of 86Rb (used to monitor K+ permeability) from preloaded cells. Cromakalim and pinacidil did not affect the catecholamine secretion induced by excessive depolarization with 56 mM K+, but inhibited that induced by moderate depolarization with 31 mM K+ in a concentration-dependent manner (1 microM-100 microM). The 31 mM K(+)-induced 45Ca2+ influx and increase in intracellular free Ca2+ concentration [Ca2+]i were also inhibited by these agents at similar concentrations to those for inhibition of catecholamine secretion. Cromakalim and pinacidil inhibited catecholamine secretion, 45Ca2+ influx and increase in [Ca2+]i induced by stimulation of nicotinic acetylcholine (ACh) receptors with carbamylcholine. Furthermore, both cromakalim and pinacidil inhibited the increase in [Ca2+]i induced by carbamylcholine in the absence of extracellular Ca2+, which is thought to be mediated by muscarinic ACh receptors. On the other hand, they did not affect catecholamine secretion induced by Bay-K 8644, Ba2+, A23187, histamine or bradykinin. These results indicate that the K+ channel openers, cromakalim and pinacidil, selectively inhibit catecholamine secretion induced by moderate depolarization or by nicotinic ACh receptor stimulation by inhibiting Ca2+ influx and increase in [Ca2+]i. Furthermore, the results suggest that these K+ channel openers-sensitive membrane K+ channels are involved in the regulation of catecholamine secretion mainly indirectly through effects on the voltage-dependent membrane Ca2+ channels.

3-Pyridinecarboxylic acid, 1,4-dihydro-2,6-dimethy↗

PubMed1994

Calcium efflux from cultured bovine adrenal chromaffin cells induced by bradykinin.

The effect of bradykinin on Ca2+ efflux from cultured bovine adrenal chromaffin cells was examined. Bradykinin enhanced the efflux of 45Ca2+ from the cells in a concentration dependent manner (10(-9)-10(-6) M). This effect was inhibited by a specific bradykinin B2-receptor antagonist, but not by a B1-receptor antagonist. Nifedipine, Co2+ and Cd2+ did not inhibit the bradykinin-stimulated 45Ca2+ efflux from the cells. 12-O-Tetradecanoyl phorbol 13-acetate, an activator of protein kinase C, also had no effect on the efflux of 45Ca2+ from the cells. The increase in bradykinin-stimulated 45Ca2+ efflux was reduced by removal of extracellular Na+. These results suggest that bradykinin stimulates Na+/Ca2+ exchange in cultured bovine adrenal chromaffin cells.

Acetylcholine↗

PubMed1994

Chromaffin cells express Alzheimer amyloid precursor protein in the same manner as brain cells.

Amyloid precursor protein (APP) 695 is remarkably expressed in the brain as compared with APP751 and APP770 which are dominant in other tissues. This study showed that human and bovine adrenal medullae dominantly expressed mRNA of APP695 as do brain nerve cells, while the adrenal cortexes expressed mRNAs of APP751 and APP770 as in other non-neural tissues. In immunohistochemistry, chromaffin cells of young rat adrenal medullae and primary cultured bovine chromaffin cells were significantly stained with a monoclonal antibody (mAb) against the common domain of the amino-terminal side of human APPs. At higher magnification, the immunostained cells revealed that APP was granularly distributed not only in the perikaryon but also in the cell processes. These results suggest that primary cultured chromaffin cells representing the state of adrenal medulla in vivo are a useful model for studying the pathophysiological functions of APPs and the mechanism of processing of APPs as a model of neuronal systems.

Adrenal Cortex↗

PubMed1994

Intravenous extended infusion of recombinant human soluble thrombomodulin prevented tissue factor-induced disseminated intravascular coagulation in rats.

This study demonstrated that intravenous infusion of recombinant human soluble thrombomodulin (rhs-TM) could inhibit disseminated intravascular coagulation (DIC) caused by 4 hr infusion of tissue factor (TF) in rats. Extended infusion of TF reduced fibrinogen and platelet counts and elevated serum FDP level. Pretreatment and coinfusion of rhs-TM could block changes of these DIC-parameters without prolongation of APTT. Heparin, which is a potent anti-DIC drug, could also inhibit these changes with extra prolongation of APTT and PT. Thus, these results suggest thrombomodulin prevent DIC less bleeding tendency than heparin.

Animals↗

PubMed1994

Osteoclastic resorption of apatite formed on apatite- and wollastonite-containing glass-ceramic by a simulated body fluid.

We immersed mirror-polished apatite- and wollastonite-containing glass-ceramic (A-W GC) disks in a simulated body fluid (SBF) for 5 days to form bonelike apatite on their surface. Neonatal rabbit bone cells were cultured on these or on plain A-W GC disks for 10, 24, and 48 h. We observed the substrates by scanning electron microscopy after treating them with pronase E plus EDTA to remove all cells except osteoclasts. Osteoclasts with a non-motile appearance formed no lacunae on the plain A-W GC, whereas on the bonelike apatite formed on A-W GC by the SBF, actively moving osteoclasts made many tracklike resorption lacunae. These were evident even after 10 h of culture and became more extensive after longer culture periods. The bonelike apatite was therefore a more suitable medium than plain A-W GC for maintaining osteoclast activity. This study demonstrated in vitro osteoclastic resorption of bonelike apatite formed on A-W GC by an SBF. It suggests that the apatite layer, through which a surface-active ceramic bonds to bone in vivo, can be resorbed by osteoclasts and subjected to bone remodeling.

Animals↗

PubMed1994

An immunohistochemical study of the extracellular matrix in oral squamous cell carcinoma and its association with invasive and metastatic potential.

The expression of extracellular matrices (ECMs) laminin (LN), type IV collagen (IV C), heparan-sulphate proteoglycan (HS-PG), fibronectin (FN), tenascin (TN), decorin and vitronectin (VN) was examined immunohistochemically in 112 primary tumours and 29 metastatic cervical lymph nodes in oral squamous cell carcinoma (OSCC). In highly invasive primary tumours, the expression of LN, IV C and HS-PG in the basement membrane along the tumour-stroma borderline and the expression of decorin and VN in the tumour stroma at the invasive site were all significantly decreased. The expression of FN and TN in the tumour stroma at the same site was markedly increased. In peritumour stroma in metastatic lymph nodes, LN, IV C, HS-PG, decorin and VN were weakly expressed, while FN and TN were strongly expressed. Thus, the staining pattern of the ECMs in the metastatic lymph nodes was similar to that in highly invasive primary tumours. Furthermore, in primary tumours of metastatic cases, the expression of LN, IV C, HS-PG, decorin and VN obviously decreased, while the expression of FN and TN increased when compared with those of the non-metastatic cases. The investigation of ECMs in OSCC was valuable in predicting tumour behaviour.

Carcinoma, Squamous Cell↗

PubMed1994

Intraarterial combined immunochemotherapy for unresectable hepatocellular carcinoma: preliminary results.

An important objective for patients with unresectable hepatocellular carcinoma (HCC) is the development of effective chemotherapy. We administered a combination of biological response modifiers and anticancer agents to 24 patients with unresectable HCC. Each case had an implanted infuser port which was connected to a catheter placed in the hepatic artery for the intraarterial (i.a.) administration of chemotherapy. The following agents were administered to each patient: recombinant interleukin-2 (800,000 JRU/day infused i.a. continuously for 6 days/week); OK-432 (5 KE injected i.a. twice in 4 weeks and i.m. three times per week); Adriamycin (10 mg injected i.a. twice in 4 weeks); cyclophosphamide (300 mg injected i.a. twice in 4 weeks), and famotidine (40 mg/day administered orally). Objective response was assessed according to tumor size measured by computed tomography and angiography before and after treatment. We observed a complete response (CR) in 4, partial response (PR) in 3, minor response (MR) in 7, no change (NC) in 7, and progressive disease (PD) in 3. The response rate (CR+PR+MR) was 58.3%. The overall 2-year survival rate was 52%. The 2-year survival rate of the responders (CR+PR+MR) was 80%, while that of the non-responders (NC+PD) was 0%. There was a significant difference between the responders and non-responders in respect to survival rate (P < 0.05). The percentages of CD25+ cells, CD56+ cells, and Leu7-CD16+ cells and NK activity in the peripheral blood showed a significant increase following the regimen. Serum levels of tumor necrosis factor alpha TNF alpha rose after the initiation of OK-432. TNF alpha levels were higher in the responders than in the non-responders. Adverse effects included high fever (all patients) and severe transient hypotension (15 patients) that was controlled by conservative therapy. Combined immunochemotherapy administered intraarterially may be a new strategy for treating unresectable HCC.

Aged↗

PubMed1994

Morphological alterations of gap junctions in phalloidin-treated rat livers.

Morphological alterations in the pattern of liver cell gap junctions were examined in phalloidin-treated rats to assess the role of gap junctions in experimental intrahepatic cholestasis. Double-labelled fluorescent staining of gap junctions and F-actin were performed using a monoclonal antibody against rat hepatocyte connexin 32 and rhodamine-phalloidin. Immunoelectron microscopy, using the anti-connexin 32 antibody, freeze-fracture replica electron microscopy, and conventional electron microscopy were also performed. In phalloidin-treated rat livers, the specific immunofluorescent staining of connexin 32 was markedly decreased in the pericentral area after 1 day of phalloidin treatment and, after 5 days of phalloidin treatment, there was a decrease in connexin 32 staining in the entire hepatic lobule. On the other hand, F-actin staining at the cell periphery and at the bile canaliculi was markedly increased in the pericentral area of the hepatic lobule after 1 day of phalloidin treatment and in the entire lobule after 5 days of treatment. Immunoelectron microscopy showed that both sides of the cytoplasmic domains of gap junctions were stained with anti-connexin 32 antibody in controls, whereas, in cholestatic rats, only one side of the cytoplasmic domain of some gap junctions was stained with anti-connexin 32 antibody after 1 or 3 days of phalloidin treatment. No gap junctions were observed after 5 days of phalloidin treatment either by freeze-fracture replica electron microscopy or by conventional electron microscopy. These results indicate that with phalloidin treatment, hepatocyte gap junctions decrease, first in the pericentral area, and finally throughout the entire lobule.

Actins↗

PubMed1994

Pathophysiologic significance of free and conjugated dopamines in congestive heart failure.

sulfoconjugated dopamine constitutes the major portion of circulating or excreted dopamine, but its physiologic significance is still unknown. To test whether conjugated dopamine serves as a source of free dopamine in response to acute stress, plasma concentrations of free and conjugated dopamine were measured during an acute exacerbation of heart failure. The plasma concentration of conjugated dopamine decreased significantly during the acute phase of heart failure, whereas that of free dopamine increased. The plasma concentration of free dopamine decreased, whereas the concentration of conjugated dopamine increased as heart failure improved. An infusion of dopamine increased the plasma concentration of conjugated dopamine, suggesting that at least part of the excess active dopamine was detoxified through conjugation. The results of these tests with both conjugated and free dopamine are interconvertible and indicate that conjugated dopamine can serve as a reservoir of active dopamine.

Adult↗