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Biomedical subjects

S Ohkubo

Publications and source records attributed to S Ohkubo.

At least 73 records · Page 4Linked to original sources

An extract of seeds from Aeginetia indica L., a parasitic plant, induces potent antigen-specific antitumor immunity in Meth A-bearing BALB/c mice.

The antitumor activity of an extract of seeds from Aeginetia indica L., a parasitic plant, was investigated. BALB/c mice, inoculated i.p. 1 x 10(5) syngeneic Meth A tumor cells, were administered 2.5 mg/kg A. indica extract i.p. every 2 days from day 0. The untreated mice died of an ascitic form of tumor growth within 21 days, whereas all the treated mice completely recovered from tumor challenge without any side-effects. The extract did not exert direct cytotoxic activity against Meth A in vitro. Mice that survived after the first challenge as a result of A. indica treatment overcame the rechallenge with homologous Meth A without additional administration of the extract. On the other hand, those mice could not survive after rechallenge with Meth 1 tumor cells, which were also established in BALB/c mice but were different in antigenicity from Meth A, suggesting the development of antigen-specific concomitant immunity in the A. indica-cured mice. In the induction phase of antitumor resistance in this system, CD4+ T cells appeared to be the main contributors, since in vivo administration of anti-CD4 mAb completely abolished such resistance. In contrast, anti-CD8 mAb administration did not influence the effect of A. indica. The importance of CD4+ T cells in antitumor immunity was again clarified by Winn assay; that is, spleen and lymph node cells depleted of CD4+ T cells in vitro prior to assay abolished antitumor activity on co-grafted Meth A tumor cells in vivo.

Animals↗

[Left ventricular diastolic function in apical hypertrophic cardiomyopathy].

To investigate left ventricular (LV) diastolic function in patients with apical hypertrophic cardiomyopathy (AHCM), we analyzed the LV cineangiograms (RAO 30 degrees) and pressures (tip manometer) in 11 patients with AHCM who had giant negative T waves on their electrocardiograms and "ace of spades" configurations on the LV angiograms. Ten patients with non-obstructive HCM (HNCM) and 10 normal subjects served as controls. LV volumes and instantaneous rates of LV volume changes were derived from frame-by-frame analyses of their LV angiograms. LV isovolumic relaxation was assessed according to the time constant (T) of LV pressure decay. LV diastolic distensibility was evaluated by plotting diastolic pressure-volume curves. There was no significant change in the LV systolic functions among these 3 groups. Compared with normals, LV end-diastolic pressure was equally elevated in AHCM and HNCM. The T of isovolumic pressure decay was significantly prolonged in AHCM and HNCM. LV early diastolic filling was maintained at the normal level in AHCM as assessed by the peak filling rate (PFR) during the rapid filling period and the time from end-systole to PFR. The LV diastolic pressure-volume relation shifted upwards in both AHCM and HNCM. In conclusion, impaired LV isovolumic relaxation and decreased diastolic distensibility, which are associated with HNCM, may also be present in AHCM.

Adult↗

[The effect of heart rate on the time constant of isovolumic relaxation of the left ventricle].

A time constant (T) of the left ventricular isovolumic relaxation, which expresses the relaxation rate of the left ventricle, may be affected by change of the heart rate. There are few clinical reports concerning the relationship between T and heart rate. We studied the relationship between T and heart rate in 10 patients with normal coronary arteries and left ventricular function. Left ventricular pressure was measured using a catheter-tip manometer during right atrial pacing, which was performed at the rate of 10, 20, 40 beats/min in addition to the baseline rate, and finished at 140 or 150 beats/min. Two measurements were made for T.1) Tw from the slope of In (pressure) against time, and 2) Tb by exponential analysis which also estimated the asymptote. As the heart rate increased, both Tw and Tb gradually shortened (Tw: from 38.5 +/- 4.9 msec at the control heart rate to 28.9 +/- 5.6 msec at 140 or 150 beats/min; Tb: from 54.7 +/- 11.4 msec at the control heart rate to 34.9 +/- 5.7 msec at 140 or 150 beats/min). These findings suggest that left ventricular isovolumic relaxation is strongly affected by change of the heart rate in man.

Adult↗

Analysis of blood spot 17 alpha-hydroxyprogesterone concentration in premature infants--proposal for cut-off limits in screening for congenital adrenal hyperplasia.

Blood Spot 17 alpha-hydroxyprogesterone (17-OHP) concentrations in neonates, especially in premature babies, were determined in relation to 1) the gestational age at birth, 2) the equivalent age of gestation at blood sampling and 3) the birth weight. The 17-OHP concentrations were found to be higher with prematurity. Accordingly, the cut-off limit in screening for congenital adrenal hyperplasia (CAH) in premature infants is proposed as 20 ng/ml. Ideal cut-off limits were set by the equivalent age of gestation at blood sampling. Cut-off limits on the basis of gestational age at birth and birth weight are also suggested, where the sampling age is not so advanced. The rate of false positivity in premature infants can be reduced by this method.

Adrenal Hyperplasia, Congenital↗

Epidermal growth factor (EGF) decreased endothelin-2 (ET-2) production in human renal adenocarcinoma cells.

Production of immunoreactive (ir-) endothelin-2 (ET-2) in renal adenocarcinoma cells, ACHN, was reduced by transforming growth factor-beta, basic fibroblast growth factor, transforming growth factor-alpha and, most strikingly, by epidermal growth factor (EGF). These growth factors did not show such inhibitory effects on the secretion of ir-ET-1 in ET-1-producing cells, indicating that the production of ET-2 and ET-1 is regulated differently by the growth factors. EGF specifically reduced the secretion of not only ir-ET-2 but also ir-big ET-2 with only a small decrease in total protein synthesis. Northern blot analysis indicated that EGF controls the ET-2-production at the transcription levels of ET-2 gene.

Adenocarcinoma↗

Effects of some radical scavengers on reperfusion-induced arrhythmias in the canine heart.

In this study, using electron spin resonance (ESR), we investigated the relation at the time of reperfusion between free radicals originating from the mitochondria of the canine myocardium and arrhythmias induced by reperfusion as well as the effect of radical scavengers on both. The left anterior descending artery was ligated just below the first diagonal branch and then reperfused for 10 minutes in 48 adult mongrel dogs. The dogs were divided into six groups consisting of: 1) control group administered no radical scavengers (n = 8), 2) SOD group (n = 6) receiving superoxide dismutase (15,000 U/kg), 3) SOD + CAT group (n = 6) receiving SOD (15,000 U/kg) and catalase (45,000 U/kg), 4) L-SOD group (n = 6) receiving liposomal-encapsulated SOD (30,000 U/kg), 5) CV-3611 (2-O-octadecylascorbic acid) group (n = 8) receiving CV-3611 (10 mg/kg), and 6) CoQ10 group (n = 6) receiving coenzyme Q10 (10 mg/kg). SOD, SOD + CAT, L-SOD, CV-3611, and CoQ10 were administered into the left atrium prior to reperfusion. The second lead of the electrocardiogram was continuously monitored during the experiment. The following results were obtained. 1) The relative intensity (RI) of the electron spin resonance signal of the mitochondria of the reperfused portion of the myocardium was smaller (p less than 0.025) in the SOD, SOD + CAT, L-SOD, CoQ10 groups (1.08 +/- 0.36, 0.92 +/- 0.19, 0.91 +/- 0.11, and 0.81 +/- 0.09, respectively) than in the control group (1.70 +/- 0.20).(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗

Expression of endothelin-2 (ET-2) gene in a human renal adenocarcinoma cell line: purification and cDNA cloning of ET-2.

It has been found that human renal adenocarcinoma ACHN cells synthesize and secrete immunoreactive endothelin (ir-ET) in the culture medium. Partial characterization of this material with reverse-phase high-performance liquid chromatography (RP-HPLC) suggested that ACHN cells synthesized only human endothelin-2 (ET-2). Isolation and characterization of this ir-ET-2 has revealed that this peptide has almost the same amino acid sequence and molecular weight as that of human ET-2 deduced from the nucleotide sequence of cloned human ET-2 gene. To delineate the precise structure of human ET-2 precursor, ET-2 cDNAs were cloned from a cDNA library constructed with mRNA derived from the ACHN cells, and the nucleotide and deduced amino acid sequences were determined. The ET-2 cDNA that has the longest open reading frame encodes prepro-ET-2 protein, consisting of 178 amino acid residues. The ET-like sequence found in the prepro-ET-1 and -ET-3 was also conserved in this prepro-ET-2. The Northern blot analysis of mRNA revealed that the transcript of the human ET-2 gene was 1.4 kb.

Adenocarcinoma↗

[A study of the determination of serum fucose concentrations].

We examined the biochemical properties of reaction products during the procedure for serum fucose determination (Dische & Shettles). The optical density at 396 nm (OD396) of the reaction products increased linearly with the increment of fucose concentrations, and was stable for at least 210-240 min at room temperature. The reaction products were destroyed with the addition of distilled water. Sephadex G-200 gel chromatography of the mixed serum samples revealed a single peak of fucose in the void volume fractions. Of interest was the presence of trace amounts of fucose detected in the low-molecular weight fractions less than 66,000, suggesting the presence of free fucose in the serum. Based on these observations, the method for serum fucose determination was modified so it was possible to use 100 microliters of the serum sample. The mean serum fucose concentration, measured by this modified method, was 561.0 +/- 191.1 mumol/l (9.1 +/- 3.1 mg/dl) in normal healthy children. In patients with idiopathic nephrotic syndrome, the values were significantly higher both at relapse and in remission than those in normal healthy children.

Chemical Fractionation↗

Specific expression of human endothelin-2 (ET-2) gene in a renal adenocarcinoma cell line. Molecular cloning of cDNA encoding the precursor of ET-2 and its characterization.

Analysis of culture medium of human renal adenocarcinoma cells ACHN by RP-HPLC suggested that the cells specifically secreted human endothelin-2 (ET-2). cDNAs encoding human ET-2 precursor were cloned from a cDNA library constructed with mRNA derived from the ACHN cells, and the nucleotide and deduced amino acid sequences were determined. The ET-2 cDNA was revealed to contain 1.3 kb and encode prepro-ET-2 protein consisting of 178 amino acid residues. The ET-like sequences found in the prepro-ET-1 and -ET-3 was conserved in this prepro-ET-2. The Northern blot analysis of mRNA suggested that the transcript of ET-2 gene was 1.4 kb. This is the first direct evidence that human ET-2 gene was expressed specifically in tumor cells.

Adenocarcinoma↗

One of the endothelin gene family, endothelin 3 gene, is expressed in the placenta.

A cDNA encoding human endothelin 3 (ET-3) precursor was cloned from a cDNA library from the placenta, and its nucleotide and deduced amino acid sequences were determined. This ET-3 cDNA was found to contain 2.3 kb pairs and encode prepro-ET-3 protein consisting of 224 amino acid residues. The putative big-ET-3 seems to consist of 42 amino acid residues. Two of the intron insertion sites were determined with information from nucleotide sequences of the cloned genomic ET-3 gene. This is the first direct evidence that the ET-3 gene is transcribed and expressed in the placenta.

Amino Acid Sequence↗

A novel peptide which stimulates adenylate cyclase: molecular cloning and characterization of the ovine and human cDNAs.

A novel neuropeptide which remarkably stimulates adenylate cyclase in rat anterior pituitary cell cultures has been recently isolated from ovine hypothalami by A. Arimura and his collaborators (Biochem.Biophys.Res.Commun.164, 567-574(1989)). This peptide was designated as PACAP38(Pituitary Adenylate Cyclase Activating Polypeptide with 38 residues). In an attempt to investigate physiological implications of PACAP38, we have succeeded in cloning the cDNAs encoding the precursor of PACAP38 from ovine hypothalamus and human testis. An ovine cDNA encodes a protein of 176 amino acids in which PACAP38 is proceeded by a putative signal peptide and a "pro"-region (107 amino acids), and followed by a Gly-Arg-Arg sequence for proteolytic processing and amidation. Deduced amino-acid sequence of human PACAP38 was completely identical to that of the ovine isolated peptide. Cloning of PACAP38 cDNAs confirms the expression of the corresponding mRNAs and the presence of this neuropeptide in ovine hypothalamus and also in human testis.

Adenylyl Cyclases↗

Complete primary structure of the human estrogen-responsive gene (pS2) product.

pS2 is a human gene whose transcription is directly triggered by estrogen in human breast cancer cells (MCF-7). We described here the complete primary structure of the pS2 gene product. The pS2 protein purified from conditioned medium of MCF-7 cells was S-pyridylethylated and digested with TPCK-trypsin. Five major fragments were obtained by reverse-phase HPLC. Amino acid sequence analysis of these tryptic peptides established that the pS2 protein comprises a 60-amino acid polypeptide. The sequence of the pS2 protein was completely identical to that deduced from the nucleotide sequence of the pS2 gene, if the signal polypeptide is excluded. Furthermore, two cDNA clones encoding an 84-amino acid precursor pS2 protein were isolated from a cDNA library which was constructed with RNA from MCF-7 cells cultured in the presence of estrogen. The nucleotide sequence of one clone (pS2B1) was identical to that of pS2 cDNA previously reported except for one nucleotide in the 3' untranslated region. The other clone (pS2B2) was longer by 73 nucleotides, at the 5' end, than pS2B1. The additional 73 nucleotides are located just upstream of the sequence of pS2B1 in the structure of the pS2 gene, indicating that the pS2 gene has two start sites for transcription. However, a mRNA molecule corresponding to pS2B1 but not to pS2B2 was detected in the cells on RNA blot hybridization analysis, indicating that one transcriptional start site is mainly used.

Amino Acid Sequence↗

[Effects of contrast media on left ventricular function. Comparison with iohexol and diatrizoate].

To investigate the effects of a low-osmolar nonionic (Iohexol: I) and a high osmolar ionic contrast media (Diatrizoate: D) on left ventricular (LV) function after left ventriculography (LVG), we studied 19 patients with coronary artery disease (I: 9 cases, D: 10 cases) during diagnostic cardiac catheterization. The first LVG was performed by injecting 40 ml of I or D at a rate of 12 ml/sec, measuring simultaneous LV pressure with a catheter tip manometer (Millar instruments). At 4 minutes after the first LVG, we repeated hemodynamic measurements and LVG. Single-plane volumes (RAO 30 degrees) were calculated every 20 msec (50 frame/sec) using the area-length method. LV systolic function was estimated by ejection fraction (EF), ratio of end-systolic pressure to end-systolic volume (ESP/ESV) and maximal (+)dP/dt. LV relaxation was assessed by the time constant (T) of LV pressure decay. LV diastolic compliance was evaluated by the diastolic pressure-volume (PV) relationship. Results. 1. LV end-diastolic pressure and volume were augmented more significantly in D than in I group, probably due to the difference of their osmolarity. 2. EF increased in D group with increase of LV preload after the first LVG, but didn't change in I group. ESP/ESV and maximal(+)dP/dt were not changed after the first LVG in both groups. 3. LV isovolumic relaxation remained unchanged with I and D. 4. LV diastolic PV relation curve shifted upward more in D than I group. We conclude that Iohexol has less influences on LV function than Diatrizoate and may be used in high risk patients.

Contrast Media↗

Selective low level of protein kinase C isozyme in a tumor promoter-dependent mouse leukemia cell line.

It was found that the activity of protein kinase C in the tumor promoter-dependent cell line (A65T) was significantly lower than that in the independent cell line (A65IND) which was mutated from the dependent cell line. On the other hand, there was no significant difference between these cell lines with regard to cAMP-dependent protein kinase activity. It was found that the maximal binding capacity of [20-3H]phorbol-12,13-dibutyrate of the tumor promoter-dependent cells is lower than that of the independent cells with similar affinities of the two cell lines. Moreover, we found that the level of immunoreactive antigen with monoclonal antibody for type III of protein kinase C in A65T cells was significantly lower than that in the A65IND cells. Thus, this selective lower level of type III of protein kinase C in A65T cells, as compared with A65IND cells means that this difference may be linked to its tumor promoter-dependent cell proliferation.

Animals↗