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Biomedical subjects

N Ochi

Publications and source records attributed to N Ochi.

At least 37 records · Page 2Linked to original sources
PubMed1998

[Clinical differences between variant and non-variant angina pectoris].

The differences in clinical characteristics were studied between variant angina pectoris with ST segment elevation during ischemic attacks and non-variant angina pectoris without ST segment elevation. Spasm provocation test was performed with either acetylcholine or ergonovine in 192 consecutive patients with vasospastic angina from January 1991 to June 1997. Thirteen patients were excluded because of insufficient data. Fifty-five patients had variant angina and 124 patients had non-variant angina. Coronary risk factors, serum cholesterol level, triglyceride level, high-density lipoprotein cholesterol level, history of syncope, the rates of second or third atrioventricular block and ventricular tachycardia or fibrillation, the incidence of organic stenosis (> or = 50%), the number of vessels with provoked spasm, the dose of acetylcholine and ergonovine used, and duration from the first appearance of chest pain were compared between the 2 groups. Patients with variant angina had more fixed stenosis (p < 0.01), required more percutaneous transluminal coronary angioplasty procedures, lower doses of intracoronary administration of acetylcholine for the induction of coronary arterial spasm and shorter duration from the first appearance of chest pains (p < 0.01) than patients with non-variant angina. However, there were no differences in other factors between the 2 groups. Variant angina pectoris has the same clinical characteristics as non-variant angina pectoris, although variant angina tends to cause higher spasmophilic activity and more fixed stenosis.

Acetylcholine↗
PubMed1998

[Absence of induced spasm by intracoronary injection of 50 micrograms acetylcholine in the right coronary artery: usefulness of 80 micrograms of acetylcholine as a spasm provocation test].

This study investigated whether the maximal dose of 50 micrograms acetylcholine for the induction of coronary spasm in the right coronary artery is adequate. The acetylcholine test was performed in 388 consecutive patients to evaluate spasm from January 1994 to December 1997. Coronary spasm in the right coronary artery was induced in 43 patients, 37 men and 6 women with a mean age of 63 +/- 8 years by intracoronary injection of 80 micrograms of acetylcholine rather than 50 micrograms. These included 15 patients (35%) with rest angina, 23 patients with ischemic heart disease other than rest angina and 5 patients (12%) with non-ischemic heart disease. Acetylcholine was injected in incremental doses of 20, 50 and 80 micrograms into the right coronary artery. Positive spasm was defined as induction of more than 90% reversible narrowing associated with either usual chest pain or ischemic electrocardiographic changes. Clinical and angiographical characteristics was studied in these patients. Fifteen (35%) patients had rest angina and 4 patients had variant angina with ST elevation in the inferior leads. Two thirds of the patients had coronary spasm in the distal portion of the right coronary artery and one third of those disclosed spasm focally. Coronary spasm was induced in 38 (15%) of 246 patients with ischemic heart disease and in 5 (4%) of 142 patients with non-ischemic heart disease. The prevalence of positive spasm in patients with ischemic heart disease was significantly higher (p < 0.01) than in patients with non-ischemic heart disease. A dose of 80 micrograms of acetylcholine, more than the maximal standard dose, might be clinically useful for the induction of spasm in the right coronary artery if coronary spasm of this artery is strongly suspected.

Acetylcholine↗
PubMed1997

Determination of lead and cadmium in food and blood by inductively coupled plasma mass spectrometry: a comparison with graphite furnace atomic absorption spectrometry.

To compare inductively coupled plasma mass spectrometry (ICP-MS) and graphite furnace atomic absorption spectometry (GF-AAS) as the method for determining lead and cadmium in the human diet and blood, 418 diet homogenate samples and the same number of blood samples were collected from Chinese and Japanese women and were analyzed by the two methods. The results showed that our ICP-MS method is precise and accurate, being comparable to the GF-AAS method established previously. The ICP-MS method is simple and fast spending only one-tenth of the time necessary for GF-AAS and allows simultaneous analyses of lead and cadmium with low detection limits. When applied to actual sample analysis, however, ICP-MS results tend to be 10-20% lower than GF-AAS results in the analysis of lead in the diet and blood and cadmium in blood. This is possibly due to some interference in ICP-MS and matrix of samples. As the ICP-MS results could be mathematically corrected to be equivalent to the GF-AAS results, we conclude that this ICP-MS method can be used as a routine analytical method for the determination of lead and cadmium in human diet and blood samples.

Blood Chemical Analysis↗
PubMed1997

The continuous and simultaneous blood flow velocity measurement of four cerebral vessels and a peripheral vessel during cigarette smoking.

There has been no consensus about the acute effect of cigarette smoking on cerebral blood flow, and the continuous change of flow in four cerebral vessel flow with peripheral flow during different kinds of cigarette smoking has not been reported until now. Our results indicate smoking increases the flow of four cerebral vessels almost at the same time and with the same pattern. Many cerebral vessels began to show increases about 10 s after commencement. In most cases, cerebral blood velocity began to decrease between 10 and 20 s after cessation. Blood flow in peripheral vessels decreases after commencement, which is thought to be the effect of nicotine. The effect of high nicotine cigarettes is greater than that of low nicotine cigarettes. Continuous and simultaneous measurement of cerebral vessels by ultrasonic Doppler is though to be the only way to establish the detailed blood flow changes during smoking.

Adult↗
PubMed1997

[Is coronary spasmodicity unchangeable?: a study with acetylcholine or ergonovine in patients with ischemic heart disease].

Fluctuations of spasmodicity have been reported in affected vessels in patients with vasospastic angina, but the incidence of spasmodicity induced by pharmacologic agents on both vessels with spasm and vessels without induced spasm has not been investigated. Repeated spasm provocation tests by acetylcholine or ergonovine were performed at 13.1 +/- 9.9 month intervals (3-50 months) in 111 vessels of 50 patients with ischemic heart disease, consisting of 19 old myocardial infarction and 31 angina pectoris, who did not undergo angioplasty or have signs of advancing atherosclerosis. Spasm was defined as present when more than 90 percent stenosis was accompanied by the appearance of usual chest pain or significant electrocardiographic changes. Ninety-six vessels (86.5%), 65 without spasm and 31 with spasm, revealed coincident responses and the remaining 15 vessels contrary reacted. The coincidence rate of spasmodicity in patients at intervals of within 24 months (90.2%) was significantly higher (p < 0.05) than that at intervals of over 24 months apart (70.4%). The spasm coincidence rate was 67.3% in the vessels with provoked spasm by either the first or second tests. Only one (0.9%) out of the 111 vessels showed obvious progressive atherosclerosis during this study. The majority of vessels showed identical spasmodicity within 2 years. In conclusion, coronary spasmodicity might remain unchanged for at least 2 years despite medication with calcium channel blockers and isosorbide dinitrate.

Acetylcholine↗
PubMed1996

Characterization, molecular cloning and expression of megakaryocyte potentiating factor.

We examined whether the conditioned media of 64 kinds of cell lines, which have been maintained by a protein-free culture system, could produce megakaryocyte potentiating (Meg-POT) activity. In these cell lines, HPC-Y5, established from human pancreatic cancer, was shown to have the highest level of activity. The megakaryocyte potentiating factor (MPF) was purified from its conditioned medium by a combination of ion-exchange chromatography, gel filtration and reversed-phase HPLC. The purified MPF showed Meg-POT activity almost equal to human (Hu) interleukin 6 (IL-6) in the presence of murine IL-3 in a colony-forming assay with mouse bone marrow cells. The molecular weight of MPF was estimated to be 33 kDa by SDS-PAGE. Glycopeptidase F digestion and amino sugar analysis of the factor demonstrated that MPF is a glycoprotein carrying at least one N-linked sugar chain. The N-terminal amino acid sequence of MPF was determined to be Leu-Ala-Gly-Glu-Thr-Gly-Gln-Glu-Ala-Ala-Pro-Leu-Asp-Gly-Val-Leu-Ala-Asn. The same or homologous amino acid sequence has not been found in known proteins, demonstrating that MPF may be a novel cytokine which has Meg-POT activity. Then, we isolated HuMPF cDNA from an HPC-Y5 cDNA library using polymerase chain reaction and plaque hybridization methods. The HuMPF cDNA encodes a polypeptide consisting of 622 amino acids, including a signal peptide of 33 amino acids, and with a deduced molecular weight of 68 kDa, although HPC-Y5 cells secrete a 33 kDa form of HuMPF. HuMPF cDNA does not show any significant homology with other known sequences. The cDNA was expressed in COS-7 and Chinese hamster ovary (CHO) cells, and Meg-POT activity was detected in their culture supernatant. The COS-7 cells secreted only a 33 kDa recombinant (r)HuMPF, however, an additional 30 kDa form was detected in the culture medium of CHO cells. The 33 kDa rHuMPF from CHO cells showed Meg-POT activity, but not the purified 30 kDa rHuMPF. The difference in structure and activity between the 33 and 30 kDa forms of HuMPF was ascribed to the existence in the 33 kDa form of the C-terminal 25 amino acid residues. The expression of MPF mRNA was examined by Northern blot analysis using labeled MPF cDNA as a probe. MPF mRNA was detected in HPC-Y5 cells, with an approximate molecular size of 2.4 kb. We also examined the expression of the MPF gene in various human tissues, and the 2.4 kb band was detected only in lung. Then, the immunohistocytochemical analysis and in situ hybridization revealed that MPF-producing cells were identified as lung macrophages. MPF may exhibit other biological activities such as regeneration of the lung tissues.

Amidohydrolases↗
PubMed1995

Molecular cloning and expression of megakaryocyte potentiating factor cDNA.

The human megakaryocyte potentiating factor (hMPF) has been previously purified from a culture supernatant of human pancreatic cancer cells HPC-Y5 (Yamaguchi, N., Hattori, K., Oh-eda, M., Kojima, T., Imai, N., and Ochi, N. (1994) J. Biol. Chem. 269, 805-808). We have now isolated hMPF cDNA from a HPC-Y5 cDNA library using polymerase chain reaction and plaque hybridization methods. The hMPF cDNA encodes a polypeptide consisting of 622 amino acids, including a signal peptide of 33 amino acids, and with a deduced molecular mass of 68 kDa, although HPC-Y5 cells secrete a 33-kDa form of hMPF. Human MPF does not show any significant homology with other previously described sequences. The cDNA was expressed in COS-7 and Chinese hamster ovary (CHO) cells, and megakaryocyte potentiating activity was detected in their culture supernatant. The COS-7 cells secreted only a 33-kDa recombinant hMPF, whereas an additional 30-kDa form was detected in the culture medium of CHO cells. The 33-kDa rhMPF purified from CHO cells showed megakaryocyte potentiating activity, but not the purified 30-kDa rhMPF. The difference in structure and activity between the 33- and 30-kDa forms of hMPF was ascribed to the existence in the 33-kDa form of the C-terminal 25 amino acid residues.

Amino Acid Sequence↗
PubMed1995

Hyperproduction of a recombinant fusion protein of Staphylococcus aureus V8 protease in Escherichia coli and its processing by OmpT protease to release an active V8 protease derivative.

The expression of a recombinant fusion protein including Staphylococcus aureus V8 protease was studied by using Escherichia coli as the host strain. When the mature V8 protease was expressed as a fusion protein with a truncated E. coli beta-galactosidase (beta-gal97S4D), we could not obtain a sufficient amount of the enzyme because of the toxicity resulting from the expressed protease activity. Synthesis of V8 protease was increased by constructing a sandwich-type fusion protein consisting of beta-gal97S4D, a V8 protease derivative with the 56 C-terminal amino acids deleted (V8 delta 56) and a truncated aminoglycoside-3'-phosphotransferase. This fusion protein was successfully produced as inactive inclusion bodies. To release the V8 delta 56 protease from the fusion protein, we developed a novel processing method using an endogeneous E. coli OmpT protease, which can recognize the dibasic amino acid residues located in the linker peptides of the fusion protein. After solubilizing the inclusion bodies with urea, the V8 delta 56 protein was automatically released from the fusion protein by the OmpT protease, which was coprecipitated with the inclusion bodies. The V8 delta 56 protease thus obtained showed the same enzymatic activity as that of the native V8 protease. We demonstrate in this study that the N-terminal prepro sequence and the C-terminal repeated sequence of this enzyme are not necessary for its enzymatic activity and protein folding.

Amino Acid Sequence↗
PubMed1995

[An 80-year-old female with pneumopericardium due to gastric perforation].

An 80-year-old woman was admitted to our hospital because of chest oppression and general malaise. On admission, auscultation of the heart revealed friction rub, so-called water-wheel like murmur. Chest X-ray film and computed tomography showed cardiomegaly and free air in the pericardium. Electrocardiogram showed low voltage with ST elevation in II, III, aVF and V1-V4. These findings suggested that the patient had pneumopericardium with pericarditis. Wenchebach type A-V block and atrial bradycardia appeared. Thus, the ventricular pacing was performed. However, patient died of multiple organ failure. The postmortempericardiography showed the gastro-pericardial fistula with hiatus hernia.

Aged↗
PubMed1994

A putative new proteinous factor negative for stromal growth. Purification and identification from endometrial carcinoma extract.

BACKGROUND: Uterine endometrial carcinoma has been reported to synthesize and secrete some putative mitogens that elicit either a positive or negative proliferation response in endometrial fibroblasts. The purposes of this study were to isolate and to identify the negative growth factor(s) from endometrial carcinoma extract. METHODS: The factor was isolated by a sequence of molecular size exclusion filtration, anion exchange chromatography, gel filtration and Affi-Gel Blue chromatography, followed by NH2-terminal amino acid sequencing. Mitogenicity was determined by [3H]thymidine incorporation into the endometrial fibroblasts. RESULTS: The purification procedure yielded a single active protein band (68 kDa). The protein, purified approximately 20,000-fold, evoked 90% inhibition of [3H]-thymidine incorporation into endometrial fibroblasts in the nanomolar range. This potent growth inhibitor is a previously unidentified protein molecule as revealed by amino acid sequences. CONCLUSIONS: Endometrial carcinoma could produce a new protein that may act as a paracrine factor to suppress the growth of its stroma endometrial fibroblasts.

Amino Acid Sequence↗
PubMed1994

A novel cytokine exhibiting megakaryocyte potentiating activity from a human pancreatic tumor cell line HPC-Y5.

Sixty-four kinds of cell lines were examined for their ability to produce megakaryocyte potentiating activity by means of conditioned media obtained from a protein-free culture system. Six human tumor cell lines were shown to produce this activity, and the cell line HPC-Y5, established from human pancreatic cancer, was shown to have the highest level of activity. The megakaryocyte potentiating factor (MPF) was purified from an HPC-Y5 conditioned medium by a combination of ion-exchange chromatography, gel filtration and reverse-phase HPLC. The purified MPF showed a megakaryocyte potentiating activity almost equal to human interleukin-6 in the presence of murine interleukin-3 in a colony formation assay with mouse bone marrow cells. The apparent molecular weight of MPF is 32,000 when determined by SDS-polyacrylamide gel electrophoresis. Glycopeptidase F digestion, and amino sugar analysis of the factor demonstrated that MPF is a glycoprotein carrying at least one N-linked sugar chain. The N-terminal amino acid sequence of MPF was determined to be Leu-Ala-Gly-Glu-Thr-Gly-Gln-Glu-Ala-Ala-Pro-Leu- Asp-Gly-Val-Leu-Ala-Asn. The same or homologous amino acid sequence has not been found in known proteins, demonstrating that MPF is a novel cytokine that has megakaryocyte potentiating activity in the murine assay system.

Amino Acid Sequence↗
PubMed1994

Spontaneous remission of diabetes arrests progression of nephropathy in streptozotocin-treated spontaneously hypertensive rats.

Although antihypertensive therapy retards the progression of diabetic nephropathy associated with hypertension, it is not known whether glycemic control reverses or arrests diabetic nephropathy under untreated hypertension. We previously reported that spontaneous remission of diabetes occurred in the neonatal streptozotocin (STZ) model of spontaneously hypertensive rats (SHR) after 28 weeks of age, whereas hypertension persisted. Thus, we studied diabetic nephropathy before and after the recovery from hyperglycemia in this model. Two-day-old male SHR were injected intraperitoneally with STZ or vehicle for control. Hypertension was developed and maintained in both STZ and control groups in a similar degree. Before the amelioration of hyperglycemia, urinary albumin excretion increased progressively in STZ-treated SHR as compared with control (24 weeks; 1.6 +/- 0.5 mg/day, 17.5 +/- 2.3 mg/day, P < 0.001), and renal and glomerular hypertrophies were seen with mesangial expansion in STZ-treated SHR. However, along the recovery from hyperglycemia, urinary albumin excretion did not increase in the STZ-treated group, while it consistently increased in the control group (52 weeks; 25.4 +/- 10.0 mg/day, 29.7 +/- 11.4 mg/day, not significant). Furthermore, there were no significant differences in renal weight, glomerular tuft area, and the incidence of glomerular sclerosis between the two groups at 52 weeks of age. This study suggests that glycemic control may be effective for diabetic nephropathy even in the coexistence of untreated hypertension.

Aging↗
PubMed1994

Polypeptide and carbohydrate structure of recombinant human interleukin-6 produced in Chinese hamster ovary cells.

A glycosylated form of human interleukin 6 (hIL-6) has been produced in Chinese hamster ovary (CHO) cells transfected with a cDNA clone for human IL-6. Recombinant hIL-6 was purified from a culture supernatant of the transfected CHO cells, and used for structural characterization. The complete amino acid sequence, composed of 185 amino acid residues, was determined and is identical to that predicted from the cDNA sequence. However, a recombinant hIL-6 species lacking two amino acid residues (Ala-Pro) from the N-terminus was also found. Two disulfide bonds are formed, between Cys45 and Cys51 and between Cys74 and Cys84. Recombinant hIL-6 carries one O-linked carbohydrate chain, and Thr139 is fully O-glycosylated. A portion of recombinant hIL-6 protein carries one N-linked sialooligosaccharide chain, and the N-glycosylation occurs at Asn46. The structure of the N-linked sugar chains was estimated by a combination of sugar mapping and glycosidase digestion. The major structure of the N-linked sugar chain predicted was of a fucosylated biantennary or triantennary complex type. Fucosylated triantennary sugar chains with one or two N-acetyllactosaminyl repeats were also found. The structure of the O-linked sugar chain was determined by 500 mHz 1H-NMR to be NeuAc alpha 2-3Gal beta 1-3(NeuAc alpha 2-6) Gal-NAcol.

Amino Acid Sequence↗
PubMed1993

[A case of multiple broncholithiasis caused by mucus retention].

The patient was a 57-year-old male with long-standing bronchiectasis who developed severe respiratory failure and died in 1991. Autopsy revealed multiple broncholithiasis in both lungs, but no calcified lymph nodes in the hilar region. Since histological examination of the broncholiths showed only stratified structures but no tissue structure, most likely cause was considered to be calcification of mucus in the bronchi. Analysis of the stone components revealed 78% calcium and 22% protein. This patient represents a case of multiple broncholithiasis caused by mucus retention, which is thought to be very rare.

Adult↗
PubMed1992

Improvement in the heterogeneous N-termini and the defective N-glycosylation of human interleukin-6 by genetic engineering.

Recombinant human interleukin-6 (IL-6), expressed in Chinese hamster ovary cells, has heterogeneous N-termini of Ala1 and Val3, as does naturally occurring IL-6. This heterogeneity is thought to be caused by difficulty in cleavage of the signal sequence. To obtain homogeneous IL-6, Pro at -1 was exchanged for Ala by site-directed mutagenesis. Alternatively, the signal sequence was replaced with that of human granulocyte-colony-stimulating factor. In both cases, the IL-6 designed to start with Ala1 was still heterogeneous, while the IL-6 designed to start with Val3 showed a homogeneous N-terminus. It is suggested that the heterogeneity of the N-terminus is caused not only by the signal sequence, but also by the succeeding sequences of the mature protein. Only a portion of recombinant human IL-6 is N-glycosylated. Asn46, being exchanged for Gln by site-directed mutagenesis, was confirmed to be partially N-glycosylated. The defective N-glycosylation was assumed to be caused by interference or tension from a disulfide bond near the N-glycosylation site. To verify this hypothesis, the Cys45 and Cys51 forming the disulfide bond were exchanged for Ser. The N-glycosylated species became predominant upon this substitution, suggesting that formation of the disulfide bond is a cause of the defective N-glycosylation.

Amino Acid Sequence↗
PubMed1992

Role of sugar chains in the expression of the biological activity of human erythropoietin.

Various deglycosylated derivatives of recombinant human erythropoietin (hEPO) were prepared and used to determine the role of the sugar chains in the expression of its biological activity in vivo and in vitro. Three N-linked oligosaccharides of hEPO have been partially or fully removed to obtain N-glycan (NG) (2)-, NG(1)-, and NG(0)-hEPO carrying two, one, and no N-linked sugar chains, respectively. The preparation lacking only O-linked sugar chain O O-glycan (OG) (0)-hEPO was also used. As de-N-glycosylation proceeded, the in vivo activity of the hormone decreased drastically, and the activity of these derivatives was correlated with the number of sialic acids bound to them. On the contrary, the in vitro activity was increased by the de-N-glycosylation; NG(0)-hEPO showed a 3-fold higher specific activity than the intact hormone. This was confirmed by binding experiments of the derivatives to target cells. The in vitro activity and the affinity also correlated with the number of sialic acids bound to the deglycosylated hEPO preparations. On the other hand, OG(0)-hEPO was as active as the intact hormone in vivo and in vitro. In conclusion, the N-linked sugar chains are not required for in vitro activity but required for in vivo activity, acting as anchors for the essential terminal sialic acids. The O-linked sugar chain has no essential role in the biological activity of the hormone in vivo or in vitro.

Amino Acids↗
PubMed1992

Continuous and simultaneous ultrasound Doppler velocimetry of the internal carotid and vertebral arteries: preliminary observations of cerebral blood flow changes with common carotid compression.

We have developed a new technique for simultaneous assessment of the four cervical cerebral arteries. Using a cervical brace with four freely adjustable Doppler probes attached, we studied four healthy young males and observed their cerebral blood flow changes during common carotid compression. The results indicated various reactivity in these subjects. We observed that carotid compression sometimes affects or stops the ipsilateral vertebral flow according to the intensity and location of compression. Also, the flow of the contralateral internal carotid artery (ICA) and vertebral artery (VA) showed remarkable increases greater than those due to ICA obliteration alone. This indicates the possibility that using only one probe during compression may often result in unknown obliteration of the ipsilateral VAs flow and could also produce other incomplete data. Moreover, with our technique, we would be able to assess the cerebral reserve more precisely and clearly in patients with carotid or vertebral occlusive diseases.

Adult↗