[Studies on human B cell activating system. I. Amplifier effects by use of two different mitogens].
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Biomedical subjects
Publications and source records attributed to T Funato.
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Thirty one healthy parturients received spinal or general anesthesia for elective cesarean section. Maternal blood pressures, maternal and fetal acid-base values, induction to delivery intervals (I-DI) and Apgar scores were determined. On spinal anesthesia, acute hydration by 6% hydroxyethyl starch solution and intramuscular injection of ephedrine were enforced prior to the block and in the left tilt position (17 degrees) oxygen inhalation immediately after the block was begun (group OII: 16 cases). As to general anesthesia, the same left tilt position was applied before the anesthesia by thiopental-0.5% halothane with-50% nitrous oxide in oxygen (group G: 15 cases). No significant decrease in maternal blood pressure was noticed, and maternal and the fetal acid-base status and Apgar scores were excellent in both groups. A positive correlation between umbilical venous pH values and 1 or 5 minutes Apgar scores was confirmed in group OII, and a negative correlation between 1 minute Apgar scores and I-DI was noticed in group G. Fetal acid-base values were not correlated with I-DI in either group, and it was postulated that placental or fetal circulation was sufficiently maintained. Biochemical status and clinical conditions in mothers and their infants are very favorable in well-conducted general or spinal anesthesia for elective cesarean section.
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Maternal blood pressures, maternal and fetal blood-gas values, induction delivery intervals (I-D intervals) and Apgar scores were determined in 60 cases of elective cesarean section under spinal anesthesia. The procedures were as follows: In the right lateral position on an operating table, the intravenous drip infusion of 6 per cent hydroxyethyl starch solution (12 ml/kg) for 30 minutes prior to spinal anesthesia and the intramuscular injection of ephedrine (0.5 mg/kg) 15 minutes before spinal anesthesia were performed. Immediately after the anesthesia, on the left semilateral position the operation was started (group S). The same studies as group S were made in cases of no reduced blood pressure without ephedrine (group E), and cases of combined use of oxygen inhalation (group O and OII). No significant reduction of blood pressure was noted in group S and O. Fetal acid-base status and Apgar scores were favorable and had no correlation with I-D intervals in any groups. It is suggested that the uteroplacental circulation in group O and E was maintained more or less smoothly than that in group S. Oxygen inhalation led to a significant increase in fetal PO2 and the improvement of 5 minutes Apgar score. On performing elective cesarean section under spinal anesthesia, it may be advisable to take the procedures of group OII (Oxygen flow: 31/min., through a nasal cannula) and to make mother respire closing their mouths.
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A sensitive and specific double antibody radioimmunoassay has been developed capable of measuring LH-RH in extracted human plasma. Thyrotropin releasing hormone, lysine vasopressin and most of LH-RH analogues did not appear to affect the assay. Hypothalamic extract and some of the LH-RH analogues produced displacement curves which were parallel to that obtained with the synthetic LH-RH. Sensitivity of the radioimmunoassay was about 3 pg per assay tube. The coefficient of variation of intraassay was 6.4%, while that of interassay was 9.6%. Exogenous LH-RH could be quantitatively extracted by acidic ethanol when varying amounts of synthetic LH-RH were added to plasma. Immunoreactivity of LH-RH was preserved in plasma until 2 hr in the cold and gradually reduced thereafter. The plasma levels in LH-RH were 20 pg/ml or less in normal adults and not detectable in children. The aged males over 60 yr and postmenopausal women showed a tendency to have higher levels of plasma LH-RH. Plasma LH-RH level was significantly higher in midcycle than in follicular and luteal stages. The disappearance rate of LH-RH from the circulation after intravenous injection could be represented as half times of 4-6 min. Between 0.2-0.4% of the injected dose was excreted into urine within 1 hr. These results indicate that the determination of LH-RH might be a useful tool for elucidating hypothalamic-pituitary-gonad interactions.
A sensitive and specific radioimmunoassay has been developed capable of measuring thyrotropin releasing hormone (TRH) in extracted human plasma and urine. All of three TRH analogues tested had little cross-reactivity to antibody. Luteinizing hormone releasing hormone, lysine vasopressin, rat growth hormone and bovine albumin were without effect, but rat hypothalamic extract produced a displacement curve which was parallel to that obtained with the synthetic TRH. Sensitivity of the radioimmunoassay was 4 pg per tube with intraassay coefficient of variation of 6.2-9.7%. Synthetic TRH could be quantitatively extracted by methanol when added to human plasma in concentration of 25, 50 and 100 pg/ml. TRH immunoreactivity was rapidly reduced in plasma at 20 degrees C than at 0 degrees C, but addition of peptidase inhibitors, FOY-007 and BAL, prevented the inactivation of TRH for 3 hr at 0 degrees C. The TRH in urine was more stable at 0 degrees C than 20 degrees C, and recovered 75 +/- 4.6% hr after being added. The plasma levels of TRH were 19 pg/ml or less in normal adults and no sex difference was observed. The rate of disappearance of TRH administered i.v. from the blood could be represented as half-times of 4-12 min. Between 5.3-12.3% of the injected dose was excreted into urine within 1 hr as an immunoreactive TRH. These results indicate the usefulness of TRH radioimmunoassay for clinical investigation.
BACKGROUND: The c-erbB-2 oncogene encodes a transmembrane tyrosine kinase receptor and its abnormal expression may be related to the prognosis of gastric cancer. Gastric cancer is relatively resistant to various drugs, including cisplatin. Cisplatin is widely used in cancer chemotherapy, but the mechanisms of drug resistance are not yet known. METHODS: We used the human gastric cancer cell lines MKN-7 and KATO-III, which express the c-erbB-2 oncogene, as a model for relative resistance to cisplatin. We investigated whether inhibition with antisense oligonucleotides against c-erbB-2 increased the sensitivity of MKN-7 and KATO-III cells to cisplatin. RESULTS: Antisense oligonucleotides for c-erbB-2 inhibited the expression of c-erbB-2 mRNA and protein and increased sensitivity to cisplatin, but not to other drugs, in MKN-7 and KATO-III cells. Cell growth was also inhibited by c-erbB-2 antisense oligonucleotides but not sense oligonucleotides. CONCLUSION: These findings indicate that c-erbB-2 expression in gastric cancer is one of the factors related to cisplatin sensitivity, and that anti-c-erbB-2 antisense oligonucleotides induced increased sensitivity to cisplatin.
Human colon cancer SW480DDP and SW620DDP cells resistant to cisplatin exhibited stronger c-myb gene expression than the parent SW480 and SW620 cells. However, cell growth rates in the cisplatin-resistant cell lines remained similar to those of the parent cells. Antisense oligonucleotides to c-myb inhibited c-myb expression and induced increased sensitivity to cisplatin in SW480DDP and SW620DDP cells, but this did not occur with the control sense oligonucleotides. In contrast, the parent cell lines were not affected by antisense oligonucleotides to c-myb. These results indicate that the c-myb gene in human colon cancer is one of the factors related to cisplatin resistance, and support the need to develop anti-cancer therapeutics based on oncogene-targeted antisense oligonucleotide technology.
Ribozymes, site-specific ribonucleases, are a new vehicle for the manipulation of gene expression. A hammerhead ribozyme designed to cleave the GUC sequence in codon 12 of activated H-ras RNA was cloned into a plasmid (pH beta Apr-1) and transfected into EJ human bladder carcinoma cells. Expression of the ribozyme dramatically reduced H-ras gene expression and inhibited growth of EJ transformants in vitro. In vivo, the H-ras ribozyme suppressed EJ cell tumorigenicity in nude mice. In contrast, the insertion of a mutant ribozyme with no demonstrable cleavage capacity into EJ cells resulted in smaller reductions in H-ras gene expression and growth inhibition while also suppressing tumorigenicity in nude mice. The ribozyme's effect on tumorigenicity was stable, as the cells were cultured for up to five months in vitro prior to injection into nude mice. These studies define a role for anti-oncogenic ribozymes as a unique class of tumor suppressing agents.
The effects of shock waves in combination with various anti-cancer agents i.e. Bleomycin (BLM), Cisplatin (CDDP) and 5-fluorouracil (5-FU) on tumor cells suspended in media containing these agents were examined. GCIY cells derived from human gastric cancer and LS 174T and SW480 cells derived from human colon cancers were used for in vitro experiments; GCIY and SW480 cells were also transplanted into nude mice for in vivo study. It was only with BLM that enhancement was evident in all three cell lines, with a degree of chemotherapeutic enhancement proportional to the amount of shock wave energy applied. Ladder formation of DNA in GCIY cells was observed only when treated with both BLM and shock waves in combination. When SW480 and GCIY cells transplanted into the backs of nude mice were treated with a combination of intravenously (i.v.) injected BLM and regional exposure to shock waves, a significant enhancement of chemotherapeutic effects was observed in terms of the tumor growth curve.
Human colon cancer cells, SW480DDP and SW620DDP, which express fos gene highly, are resistant to cisplatin treatment. We prepared a hammerhead ribozyme to selectively cleave fos mRNA and revealed that the for ribozyme significantly suppressed the expression of fos gene in resistant cells in vitro. The fos ribozyme which was transfected into the implanted tumor cells resistant to cisplatin, had also the ability to reduce the expression of fos gene in vivo, and reversed cell sensitivity to cisplatin. These results reinforce the potential role of anti-oncogene ribozymes in dealing with the problem of drug resistance, with new possible implications for gene targeting therapy.