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Biomedical subjects

R Takaki

Publications and source records attributed to R Takaki.

At least 55 records · Page 3Linked to original sources
PubMed1989

Serum-free culture of insulin-secreting clonal cells from a hamster insulinoma.

We experimented with a wide range of serum-free media to find the best one for culturing insulinoma cells from the Syrian golden hamster, cell line In-R1-I10. Optimum cell growth came with a mixture of equal proportions of Dulbecco's modified Eagle's medium and Ham's F-12, supplemented with 10(-6) M insulin, 10 micrograms/ml transferrin, and 10(-9) M triiodothyronine (what we labeled DF-ITT medium). In addition to testing different varieties of basal media, we also experimented with different concentrations of known stimulants of cell proliferation, including transferrin, ferrous sulfate, insulin, epidermal growth factor, triiodothyronine, hydrocortisone, monoethanolamine, prolactin, proteose peptone, and selenium. Cells cultured in DF-ITT medium grew as well as those in serum-containing medium for 94 consecutive generations. Their insulin secreting capacity was maintained. The substitution of epidermal growth factor (10 ng/ml) for the insulin did not reduce either the growth rate or the insulin secreting capacity of the culture cells.

Adenoma, Islet Cell↗
PubMed1989

Protective role of potentially lethal damage repair in the neoplastic transformation of Balb/c 3T3 cells treated with N-methyl-N'-nitro-N-nitrosoguanidine.

To determine the role of repair of potentially lethal damage (PLD) in the initiation process of neoplastic transformation, Balb/c 3T3 cells treated with N-methyl-N'-nitro-N-nitrosoguanidine (MNNG) were temporarily exposed to conditioned medium obtained from density-inhibited Chinese hamster cell cultures, as a post-treatment for the induction of PLD repair. With or without this exposure, cell survival and transformation frequencies were simultaneously determined by colony-formation and focus-formation assays, respectively. Temporary exposure to conditioned medium resulted in a 20-30% increase in cell survival compared with no exposure. Post-treatment with conditioned medium resulted in a 60-65% reduction in transformation frequencies. At the molecular level, the repair of MNNG-induced single-strand breaks of DNA occurred much more rapidly in conditioned medium. These data suggest that PLD repair reduces the in vitro neoplastic transformation through excision repair operative during the cessation of DNA replication. Thus, PLD repair appears to be preventive against neoplastic fixation in initiation of neoplastic development.

Animals↗
PubMed1989

[Three cases of Torsades de pointes].

Torsades de Pointes is an atypical ventricular tachycardia, a characteristic sinusoidal twisting of QRS complex around the base line, first described by Dessertenne et al in 1966. Since their original description, many literatures including both reviews and case reports have been issued. However, the exact causes, mechanism and its true relation to usual ventricular tachycardia and fibrillation remain to be elucidated. In the present report, three cases of Torsades de Pointes (TDP) were described. All these patients revealed a prolonged QT and QTc interval as has been recognized at the occasion of TDP attacks together with low serum potassium concentration. In addition, one had hypertrophic cardiomyopathy (case 1), cholelithiasis (case 2) and uterus cancer (case 3), respectively. Regarding to the treatment, direct current counter shock was applied to terminate the TDP in all 3 cases, the correction of low serum potassium concentration was also done in each case. Lidocaine was ineffective in case 3 to terminate the TDP, while verapamil was effective in case 3 to convert TDP to normal sinus rhythm. In case 2, no antiarrhythmic drugs were applied, however, cardiac massage and normalization of serum potassium concentration succeeded to recover normal sinus rhythm. The diversity of the causes and treatments of TDP in these three cases was discussed in relation to the genesis of TDP.

Adult↗
PubMed1988

[Studies on serum ketone bodies in patients with hyperthyroidism].

In order to investigate effects of thyroid hormone on ketone bodies metabolism, fasting levels of serum ketone bodies, serum free fatty acids (FFA), serum insulin (IRI), plasma glucagon (IRG) and plasma glucose were examined in 29 untreated patients with hyperthyroidism and 20 healthy subjects. In 21 patients the levels of serum ketone bodies were re-examined when euthyroidism was achieved after treatment. In all of healthy subjects and 17 patients changes in the levels of serum ketone bodies after oral glucose load were examined. The results were as follows: 1). Fasting levels of serum FFA and total ketone bodies (TK), acetoacetate (AcAc), 3-hydroxy-beta-butylate (3OHBA), ratio of 3OHBA to AcAc in the patients were significantly higher than those in healthy subjects. The levels of IRI, IRG or ratio of IRG to IRI in the patients were not different from those in healthy subjects. In the patients, the fasting level of TK was significantly correlated with the level of FFA. 2). After oral glucose load the levels of TK and FFA in the patients decreased gradually. 3). The fasting levels of TK and FFA in the patients decreased when euthyroidism was achieved after treatment. It was suggested that the fasting levels of serum ketone bodies in patients with hyperthyroidism elevated probably due to activated lipolysis.

Blood Glucose↗
PubMed1988

Ultrastructural and functional studies of pancreatic B cells in Wistar rats treated with immunotherapeutic doses of cyclosporin.

Cyclosporin (CYA) was administered to Wistar rats at immunotherapeutic doses (20 mg/kg) and the functional and morphological effects on pancreatic B cells were observed. Serum CYA levels were kept in the range that is often encountered in clinical immunotherapy in humans. The treated rats had slightly elevated plasma glucose levels; on treatment days 6 and 13, their serum immunoreactive insulin and pancreatic insulin contents were low. Plasma immunoreactive glucagon levels were slightly elevated during treatment. Glucose tolerance was impaired on day 13. These functional abnormalities disappeared 2 weeks after drug withdrawal. B cells showed ultrastructural evidence of CYA toxicity, including cytoplasmic degranulation, nuclear inclusions, and cisternal dilatation of both the rough endoplasmic reticulum and the Golgi apparatus. These changes were observed on the third day of treatment and became more pronounced on the 6th and 13th days of treatment. By the second week after drug withdrawal, most of the B cells presented a normal ultrastructure. We confirmed that a dose of CYA similar to that used in clinical immunotherapy in humans produces ultrastructural disturbances in the pancreatic B cells of Wistar rats after relatively short-term administration.

Animals↗
PubMed1988

Characterization of secretory responses of a glucagon-producing In-R1-G9 cell line.

The In-R1-G9 cell line is one of the clones derived from the In-111-R1 hamster insulinoma cell line and produces glucagon. The secretory responses of In-R1-G9 cells were further examined to characterize the nature of the cells. Vincristine had no effect on glucagon secretion and colchicine enhanced glucagon secretion slightly after a short incubation. Two calmodulin inhibitors, trifluoperazine and chlorpromazine, did not affect glucagon secretion. Monensin at 10(-8) M suppressed glucagon secretion by 50%. Secretion of glucagon was calcium-dependent. The addition of A23187 to the incubation medium resulted in a 180% increase over control for 1 h and calcium deprivation from the medium suppressed glucagon secretion markedly. Theophylline, a phosphodiesterase inhibitor, caused a 230% increase in glucagon secretion. An experiment using cycloheximide suggested that newly synthesized glucagon appears in the medium at 30 min. This cell line should be useful for various experiments in many fields of research.

Adenoma, Islet Cell↗
PubMed1988

A case of arrhythmogenic right ventricular dysplasia with prominent late potential.

A case of arrhythmogenic right ventricular dysplasia with frequent episodes of ventricular tachycardia (VT) in a 57-year-old man is reported. His electrocardiogram showed a normal axis and inverted T waves in V1-V3 during sinus rhythm and a left bundle branch block pattern during VT. The VT could be terminated by either intravenous administration of lidocaine or procainamide or by cardioversion. Echocardiogram and right ventriculography showed reduced wall motion of the apex and infundibulum of the dilated right ventricle. No evidence of ischemic heart disease was found by left ventriculography, coronary arteriography and RI scintigraphy. A prominent late potential following the QRS complex was recorded by averaging and magnifying the surface electrocardiogram. The combination of oral mexiletine and disopyramide partially suppressed the VT attacks and relieved the subjective symptoms, however the late potential remained almost unchanged.

Cardiac Pacing, Artificial↗
PubMed1988

Aggregate size and viscoelastic property of a highly concentrated suspension.

Flow of Latex suspension with high concentration between plate glasses was observed through an optical microscope. Size of aggregates of Latex particles was estimated by measuring area of a region without particles behind an obstacle in the flow region (this technique had been developed by one of the authors). At the same time, viscosity and elasticity of the suspension were measured by a cone-plate viscometer, and relations between the aggregate size and these properties were obtained. A simple model based on the Stokes law of resistance and the Maxwell model for viscoelastic materials is proposed to explain these relations. The theoretical results agree with the present experiment.

Elasticity↗
PubMed1987

The complete cancellation of abnormal Q waves due to an old anteroseptal infarction following subsequent acute posterior myocardial infarction.

A rare case of multiple infarction is described. An abnormal Q wave due to an old anteroseptal infarction was completely masked by a new contralateral posterior myocardial infarction. Coronary angiogram revealed multiple stenoses of segments 1 (100%), 7 (99%), 12 (90%) and 13 (99%). Left ventriculography also showed akinesis of segments 2, 3 4 and 6, and reduced wall motion in segments 1, 5 and 7. These findings, together with electrocardiographic changes from before the present attack reinforce the above interpretation.

Aged↗
PubMed1986

Isolation of glucagon-secreting cell lines by cloning insulinoma cells.

Six glucagon-secreting cell lines designated as In-R1-G1, -G3, -G7, -G9, -G10, and -G11 were isolated from insulinoma cells (In-111-R1) by single cell cloning. A small amount of insulin was also detectable in the incubation medium when hormone secretion was stimulated by the addition of arginine or theophylline. These cell lines grew as monolayers and the population doubling times varied from 16.8 to 28.8 h. Karyologically these clones were aneuploid and the modes of chromosome numbers were 61 to 70. Electron microscopic examination of one of these clones showed that these cells contained moderately developed Golgi apparatus and a few secretory granules, which more or less resembled alpha-cell granules. By gel filtration study of the incubation medium, glucagon and glucagonlike material were eluted. The molecular weight of the latter was approximately 9000, which suggested the concomitant secretion of proglucagon into the medium. The levels of secreted glucagon in basal state were 0.3 to 3.0 ng/10(6) cells/2 h. Glucagon secretion was markedly enhanced in the presence of amino acids. Glucagon secretion increased slightly in the presence of high concentration of glucose in Hanks' balanced salt solution; however it was not affected by the varying concentrations of glucose when the cells were incubated in complete media with amino acids. Glucagon secretion was also stimulated by the addition of theophylline. These clonal cell lines seem to provide a useful tool for investigating the mechanism of glucagon secretion.

Adenoma, Islet Cell↗
PubMed1986

Effect of phorbol esters on glucagon secretion from a glucagon-secreting clonal cell line. Synergistic effects of A23187 and theophylline.

The cell line In-R1-G9 is one of the clones from the hamster insulinoma cell line, In-111-R1, and it produces glucagon. Phorbol esters markedly enhanced glucagon secretion and the stimulatory effect was found to be correlated to their biological activity as tumor promoters. At a concentration of 200 nM, 12-O-tetradecanoylphorbol 13-acetate (TPA) stimulated glucagon secretion 13-fold more than the control in 10 min. The effect of TPA was not influenced by actinomycin D, cycloheximide, colchicine or vincristine. Depletion of calcium from the incubation medium inhibited TPA-induced glucagon secretion by approximately 50% and dibucaine also suppressed glucagon secretion to 67.4%. An addition of A23187 to TPA induced 150% enhancement over the TPA-stimulated glucagon level, and the maximum secretory response was observed when the cells were stimulated with the simultaneous addition of TPA, A23187 and theophylline.

Adenoma, Islet Cell↗
PubMed1986

Combined effects of glucose and oxygen concentrations upon the ultrastructure of cultured islet cells and insulin accumulation in culture media.

Rat pancreatic islets cultured for 6 days at 100 or 500 mg/dl glucose and 20 or 7% O2 were examined electron-microscopically, and insulin accumulation in the culture media was assayed immunologically. In the islets cultured at 500 mg/dl glucose and 20% O2, B cells exhibited hypertrophy of granular endoplasmic reticulum and Golgi apparatus, an abundance of free ribosomes, degranulation and the margination of secretory granules. In islets cultured at 500 mg/dl glucose and 7% O2, B cells exhibited dilatation of endoplasmic reticulum cisternae and dominance of Golgi vesicles in addition to the above-mentioned changes. These changes, together with the correlated data on insulin accumulation, are discussed with special reference to the effects of glucose and oxygen upon the synthesis and release of insulin in B cells.

Animals↗
PubMed1985

Enhancement of erythroid progenitor cell growth in medium conditioned by a human cancer cell line, KONT.

Erythroid-potentiating activity (EPA) was detected in culture medium conditioned by a human cancer cell line (KONT) that produces colony-stimulating activity (CSA), using erythroid colony formation in vitro. EPA in the medium conditioned by the KONT cells (KONT-CM) was markedly heat stable. After treating KONT-CM at 80 degrees C for 30 min, 30% EPA remained, while CSA was completely inactivated. Both EPA and CSA appeared in approximately the same fractions of the gel filtration, indicating a molecular weight of approximately 30,000 daltons. EPA bound partially to Concanavalin-A Sepharose, whereas CSA almost did not bind. Our results indicate that EPA can be separated from CSA based on heat stability and binding to Concanavalin-A Sepharose.

Cell Division↗