Biomedical subjects

S Inoue

Publications and source records attributed to S Inoue.

At least 109 records · Page 6Linked to original sources

PubMed1991

Molecular cloning and sequencing of the epidermal cell differentiation inhibitor gene from Staphylococcus aureus.

We recently purified to homogeneity a protein inhibiting differentiation of cultured keratinocytes from extracellular products of Staphylococcus aureus, and named it epidermal cell differentiation inhibitor (EDIN). In the present study, we isolated and sequenced the structural gene coding for EDIN from Staphylococcus aureus E-1 using oligonucleotide probes on the basis of the partial amino acid sequence of the purified EDIN. DNA sequencing of the cloned DNA revealed an open reading frame encoding 247 amino acids as a precursor of EDIN, which included an NH2-terminal signal sequence of 35 amino acid residues. Processing of this precursor produces a mature EDIN protein composed of 212 amino acids with a calculated Mr of 23,782. The EDIN shared 35% amino acid homology with the ADP-ribosyltransferase C3 of Clostridium botulinum. These results with biological properties of EDIN described previously indicate that EDIN is a novel protein.

Amino Acid Sequence↗

PubMed1991

Amino acid sequences of the two subunits of a phospholipase A2 inhibitor from the blood plasma of Trimeresurus flavoviridis. Sequence homologies with pulmonary surfactant apoprotein and animal lectins.

Phospholipase A2 inhibitor (PLI), purified from the blood plasma of the Habu snake (Trimeresurus flavoviridis), was separated into two distinct subunits, PLI-A and PLI-B. These subunits were shown to be glycoproteins with molecular weights of around 21,000-22,000. When they were deglycosylated chemically with trifluoromethanesulfonic acid, the molecular weights were found to be 17,000. Their amino acid sequences were determined by alignment of peptides obtained by lysyl endopeptidase digestion and Staphylococcus aureus V8 protease digestion. PLI-A and PLI-B were each composed of 147 amino acid residues with one residue, Asn103, being for N-linked glycosylation, and the molecular weights of their protein portions were calculated to be 16,368 and 16,408, respectively. Each subunit contained four cysteine residues, all of which exist in disulfide linkages (Cys64-Cys141 and Cys119-Cys133). The sequences of PLI-A and PLI-B showed 89.9% homology to each other. When the sequences were compared with those of lipocortins, no significant homologies were detected. But the sequences were significantly homologous to those of COOH-terminal carbohydrate recognition portions of pulmonary surfactant apoprotein and animal lectins.

Amino Acid Sequence↗

PubMed1991

Time-resolved fluorometric sandwich immunoassay for human growth hormone in serum and urine.

A specific and sensitive time-resolved fluorometric sandwich immunoassay for human growth hormone (hGH) in serum and urine is described. A monoclonal anti-hGH IgG1 (5802)-coated polystyrene ball was incubated with serum or dialyzed urine and subsequently with europium ion-labeled monoclonal anti-hGH IgG1 (5801). Europium ion bound to the polystyrene ball was measured by time-resolved fluorometry. The detection limit of hGH was 0.3 pg/tube, which was 15-fold higher than that by sandwich enzyme immunoassay using horseradish peroxidase as label. The assay range of serum hGH was 15-15,000 ng/liter using 20 microliters of serum. The assay range of urinary hGH was 2-2,000 ng/liter using 150 microliters of dialyzed urine. The detection limits of hGH in serum and urine by this immunoassay were satisfactory for measuring hGH levels in serum and urine of healthy children and in serum of healthy adults and higher levels but not in urine of healthy adults and in serum and urine of patients with hGH deficiency.

Adolescent↗

PubMed1991

Effect of ecdysterone on histamine release from rat peritoneal mast cells.

Ecdysterone dose-dependently inhibited anti-IgE-induced histamine release from mast cells. Moreover, the rate and extent of histamine release from mast cells induced by Concanavalin A (Con A) are significantly diminished in samples incubated with ecdysterone. Ecdysterone inhibited both the initial and gradual rise in fluorescent response by anti-IgE and Con A. The effects of ecdysterone on the fluorescence response was correlated with the inhibition of histamine release. These results suggest the possibility that the inhibition of histamine release from rat mast cells by ecdysterone might be due to inhibition of Ca2+ mobilization from intracellular Ca2+ storage.

Animals↗

PubMed1991

Pentosome--a new connective tissue component--is a subunit of amyloid P.

A new minute connective tissue structure, referred to as "pentosome", has been investigated by electron microscopy and its nature has been examined by immunoperoxidase tests. Pentosomes are 3.5-nm wide, particulate structures that have been observed in the posterior chamber of the eye, the connective tissue spaces of the mouse foot-pad and the matrix of the mouse EHS tumor. They are usually found in the vicinity of microfibrils whether they are free or associated with elastic fibers. They tend to be organized into groups forming a three-dimensional semi-crystalline lattice at 10-nm intervals, but are connected by fine filaments. At high magnification, pentosomes appear as hollow structures composed of two parallel pentagons, which respectively measure 2.7 and 3.5 nm, and are held together by a cross-bar. A series of immunoperoxidase tests has only shown antigenicity against a serum protein, the amyloid P component. However, pentosomes are only about one-third the size of the 8.5-nm wide, disk-like segments of the amyloid P molecule. Since they could be subunits of these molecules, such subunits were prepared and compared with pentosomes; they appeared to be identical. It is concluded that the pentosomes found in connective tissue are AP subunits.

Animals↗

PubMed1991

Repair of an aortoiliac aneurysm associated with chronic disseminated intravascular coagulopathy--a case report.

We report herein an unusual case of a 77 year old man who developed chronic disseminated intravascular coagulopathy (DIC) caused by an aortoiliac aneurysm. After preoperative heparin therapy, the aneurysm was successfully replaced with a prosthetic graft despite heavy blood loss. Moreover, the infusion of platelet concentrate and autotransfusion proved helpful during the operation and his hypofibrinogenemia and thrombocytopenia became normalized immediately after surgery. Thus, although preoperative heparin therapy can help control DIC, surgical repair of the aneurysm is the definitive treatment in this kind of case.

Aged↗

PubMed1991

Surgical treatment of a huge hepatic artery aneurysm--a case report.

A debilitated 68 year old woman with an epigastric mass previously identified as a hepatic artery aneurysm (HAA) by selective angiography, developed acute epigastric pain radiating to her back. Impending rupture was diagnosed, and the patient successfully underwent surgical repair using a prosthetic patch to close the orifice of the aneurysm. HAA is uncommon, but the risk of rupture is great, being 44 per cent and the mortality rate high, being 35 per cent. Abnormal ultrasonography and CT may suggest the diagnosis, but selective arteriography is definitive. Surgical treatment is indicated in most patients, although embolization is appropriate for intrahepatic aneurysm and extremely poor risk patients.

Aged↗

PubMed1991

Surgical experience with nonparasitic cysts of the liver--the characteristics and constituents of cyst fluid.

Fifteen patients with nonparasitic hepatic cysts underwent surgery to relieve abdominal complaints, at the First Surgical Department, Tokyo University Hospital between 1966 and 1989. Total removal of the cyst was carried out in 14 of the patients, while puncture and aspiration only were performed in the other patient in whom a cyst had ruptured spontaneously, after which hemoperitoneum was observed at emergency laparotomy. The size of the cysts varied from 7-19 cm with a mean of 12.4 cm. Histological study revealed a congenital cyst in 14 patients and a cystadenoma in one. The epithelium lining of the internal surface of the congenital cysts was highly atrophic and desolated and often disappeared. Examination of the cyst fluid revealed the same concentration of electrolytes as in the serum. Hepatic enzyme levels were usually lower than in the serum, but lactic dehydrogenase and aminotransferase levels were elevated in a few cases. Tumor marker levels of the cyst fluid were higher than those in the serum in four of five patients examined.

Adult↗

PubMed1991

Age-related decrease in plasma growth hormone: response to growth hormone-releasing hormone, arginine, and L-dopa in obesity.

Aging is associated with a reduction in plasma growth hormone (GH) secretion in non-obese subjects. To determine whether or not age-related changes in plasma GH secretion exist in obese subjects, we measured (a) plasma GH response to growth hormone-releasing hormone (GRH; 1 microgram/kg body wt), arginine (0.5 g/kg body wt), L-dopa (500 mg), and (b) plasma glucose, insulin, and free fatty acids (FFAs) in 26 fasted obese subjects of various ages ranging from 16 to 71 years. Only subjects with a body mass index (BMI; kg/m2) between 30.0 and 39.0 were studied. Six subjects were adolescents, 9 were in their 20s, and 11 were 30 years or older. The mean peak levels of plasma GH in response to GRH, arginine, and L-dopa in obese subjects were 11.3 +/- 2.1, 21.9 +/- 4.4, and 5.2 +/- 0.3 ng/mL in adolescents, 8.2 +/- 1.6, 9.1 +/- 1.5, and 3.1 +/- 0.6 ng/mL in those in their 20s, and 4.5 +/- 0.4, 7.3 +/- 1.4, and 2.8 +/- 0.3 ng/mL in those 30 years or older, respectively, showing a significant decrease in peak GH level with advancing age (P less than .05 to P less than .01).(ABSTRACT TRUNCATED AT 250 WORDS)

Adolescent↗

PubMed1991

Impaired biphasic insulin release in mildly diabetic rats bearing a chronic portal vein catheter.

Using a new technique of chronic portal vein catheterization in freely moving rats, we investigated the insulin release to intravenous (IV) glucose (0.5 g/kg) injection in mildly diabetic (35 mg/kg streptozotocin, IV) rats. In nondiabetic rats, plasma insulin of the portal vein showed a clear biphasic release pattern, which peaked within two min after glucose injection, reached a nadir between three and six min, then began to rise to a second peak between 7 and 10 min, and reached a second nadir at 14 min. In mildly diabetic rats, a biphasic insulin release was evident, but both the first and the second phase insulin releases were impaired. In spite of this impaired insulin release, glucose intolerance was mild. Simultaneous blood sampling from the portal and peripheral veins after IV glucose injection revealed that diabetic rats showed diminished hepatic insulin extraction. These results suggest that the biphasic insulin release to glucose is impaired in mildly diabetic rats, but diminished hepatic insulin extraction contributes at least to keeping glucose tolerance mild.

Animals↗

PubMed1991

DHT formation and degradation in cultured human skin fibroblasts: DHT accumulation in the genital skin.

The conversion of testosterone to dihydrotestosterone (DHT) by 5 alpha-reductase and the interconversion between DHT and 5 alpha-androstane-3 alpha, 17 beta-diol (3 alpha-diol) by 3 alpha-hydroxy-steroid oxidoreductase (3 alpha-HSOR) were studied in fibroblasts derived from the genital skin of 22 males and 6 females, and from the nongenital skin of 19 males and 9 females with normal gonadal function. The formation of DHT from testosterone (5 alpha-reduction) was significantly greater in fibroblasts from genital skin than in those from nongenital skin in both males (2.15 +/- 1.43 vs 0.81 +/- 0.46 pmol/mg protein/h, mean +/- SD, P less than 0.001) and females (2.52 +/- 1.99 vs 0.69 +/- 0.18, P less than 0.01). Furthermore, DHT formation from 3 alpha-diol (3 alpha-HSOR oxidation) was also significantly greater in genital skin fibroblasts than in nongenital skin fibroblasts of males (5.47 +/- 3.37 vs 2.52 +/- 1.74 pmol/mg protein/h, P less than 0.01). However, the degradation of DHT to 3 alpha- and/or 3 beta-diol (3 alpha- and/or 3 beta-HSOR reductions) was not different between genital and nongenital skin fibroblasts of either males or females. Respective ratios of DHT formation to DHT degradation (5 alpha-reduction/3 alpha-HSOR reduction, 3 alpha-HSOR oxidation/3 alpha-HSOR reduction) were also significantly greater (P less than 0.002) in genital skin fibroblasts than in nongenital skin fibroblasts of males. On the other hand, both DHT formation and degradation were not different between male and female genital skin fibroblasts. These results suggest that the increased production of DHT in genital compared to nongenital skin results from increased 5 alpha-reduction and 3 alpha-HSOR oxidation.

3-Oxo-5-alpha-Steroid 4-Dehydrogenase↗

PubMed1991

A 24-week study of the microleakage of four retrofilling materials using a fluid filtration method.

The microleakage after retrofillings of amalgam, amalgam with cavity varnish, silver-containing glass ionomer cement, and intermediate restorative material was compared in vitro. Thirty-six extracted human incisors and canines were instrumented, obturated with gutta-percha without sealer, subjected to apicoectomy, and retrofilled with the materials described above. After removal of the gutta-percha filling, the microleakage from both directions (apically and coronally) was measured quantitatively and longitudinally for 24 wk after filling by a fluid filtration technique. All four retrofilling materials revealed some apical and coronal leakage at all time periods. The amalgam group showed statistically significant (p less than 0.001) apical leakage at 1.5 h. The use of cavity varnish significantly reduced the apical leakage of the amalgam group at 1.5 h. The silver-containing glass ionomer cement and intermediate restorative material groups showed significantly (p less than 0.05) less coronal leakage compared with the amalgam group at 1.5 h. Scanning electron microscopy of selected samples showed marginal defects of approximately 5 microns between the root dentin and the retrofilling material.

Cermet Cements↗

PubMed1991

Kinetics of the hydrolysis of monodispersed and micellar phosphatidylcholines catalyzed by a phospholipase C from Bacillus cereus.

The phosphatidylcholine-hydrolyzing phospholipase C, so-called "phospholipase C" (PLC), was isolated from the culture of Bacillus cereus strain IAM 1208. The amino-acid composition and partial N-terminal sequence of the purified enzyme were in good agreement with those expected from the nucleotide sequence for a PLC of strain ATCC 10987 [Johansen et al. (1988) Gene 65, 293-304]. The chain-length dependence of kinetic parameters for the PLC-catalyzed hydrolysis of monodispersed short-chain phosphatidylcholines (diCNPC, N = 3-6) was studied by a pH-stat assay method at 25 degrees C, pH 8.0, and ionic strength 0.2 in the presence of saturating amounts of Zn2+ (0.1 mM). The result was compared with those for snake venom phospholipases A2 [Teshima et al. (1989) J. Biochem. 106, 518-527]. It was found that the interaction of the PLC with the head group of the substrate molecule is very important for the binding. The pH dependences of kinetic parameters for the hydrolysis of monodispersed diC5PC and mixed micelles of diC16PC with Triton X-100 were also studied under the same conditions. An ionizable group, whose pK value is perturbed from 7.77 to 8.30 by substrate binding, was found to be essential to the catalysis. This group was tentatively assigned to His 14 on the basis of the results on X-ray crystallographic and chemical modification studies [Hough et al. (1989) Nature 338, 357-360 and Little (1977) Biochem. J. 167, 399-404].(ABSTRACT TRUNCATED AT 250 WORDS)

Amino Acids↗

PubMed1991

Kinetics of the hydrolysis of monodispersed dihexanoylphosphatidylcholine catalyzed by bovine pancreatic phospholipase A2: roles of Ca2+ binding and ionizations of amino acid residues in the active site.

Phospholipases A2 (PLA2s) are classified into two groups, I and II, according to differences in the polypeptide-chain length and intramolecular disulfide bondings. The hydrolysis of monodispersed 1,2-dihexanoyl-sn-glycero-3-phosphorylcholine (diC6PC), catalyzed by bovine pancreas PLA2 (Group I) was studied at 25 degrees C and ionic strength 0.2, and the initial velocity data were analyzed by means of the Michaelis-Menten equation. The Michaelis constant, Km, was found to be practically independent of Ca2+ concentration and also of pH value. The latter result indicates no participation of the ionizable groups in the active site in the substrate binding. The pH-dependence curve of the logarithm of the catalytic center activity, kcat, obtained in the presence of a practically saturating amount of Ca2+, showed three transitions ascribable to the participation of three ionizable groups with pK values of 5.00, 8.40, and 9.50. The respective groups were tentatively assigned to the catalytic group His 48, the N-terminal alpha-amino group, and invariant Tyr 52, which is located in close proximity to the imidazole ring of His 48. Deprotonation of His 48 and protonation of Tyr 52 were shown to be essential to the catalysis. The importance of the ionization state of the alpha-amino group was also indicated.(ABSTRACT TRUNCATED AT 250 WORDS)

Amino Acids↗

PubMed1991

Subepicardial microischemia formation induced by epicardial application of endothelin-1.

The effects of endothelin-1 (ET-1), applied topically on the epicardium, on the coronary microcirculation of the beating canine heart were investigated in situ. ET-1 (1, 10, 100, and 1,000 pmol) induced a dose-dependent elevation of the ST segment in the epicardial ECG (n = 5). After application of ET-1 (100 pmol), the beating hearts were rapidly cross-sectioned and freeze-clamped in 120 ms using a specially developed device (n = 6). By NADH fluorescence photography of the cross-sectioned frozen sample, an increased fluorescent area in the subepicardium was clearly observable. The fluorescent dye injected from the left atrium was negative in the NADH fluorescent area. It can be concluded that ET-1, when administered extravascularly, induces severe vasoconstriction and myocardial ischemia in the microcirculation of the beating canine heart.

Administration, Topical↗

PubMed1991

Successful treatment of a hyperprolactinemic infertile woman with a pituitary microadenoma using hachimijiogan.

A hyperprolactinemic infertile woman with a pituitary microadenoma, who was resistant to bromocriptine as a method of bringing on pregnancy, succeeded in having a normal pregnancy and delivery with Hachimi-jio-gan treatment. This paper reports that the patient did not succeed in pregnancy in spite of 16 months of bromocriptine treatment, though her prolactin level gradually decreased and continued to be between 40-60 ng/ml with Hachimi-jio-gan treatment. Although the level of prolactin of the patient did not normalize, she, nevertheless, succeeded in having a normal pregnancy and delivery.

Adenoma↗

PubMed1991

Hepatocellular carcinoma: treatment with a combination therapy of transcatheter arterial embolization and percutaneous ethanol injection.

In an attempt to obtain complete tumor necrosis in large hepatocellular carcinoma (HCC) lesions, the authors studied the clinical and histologic findings of a new combination therapy, percutaneous ethanol injection (PEI) with transcatheter arterial embolization (TAE) (pretreatment with TAE and subsequent PEI) in 15 patients with a single, large (3.0-9.0 cm in diameter), encapsulated lesion of HCC. Two weeks after TAE, PEI was performed under ultrasound guidance. A total of four to 11 injections were administered at a rate of one injection twice a week. During the follow-up period (range, 7-23 months), all lesions were reduced in size and no evidence of HCC was present at contrast material-enhanced computed tomography or angiography in nine of 11 patients who did not subsequently undergo surgery. Six patients had a follow-up of 1 year or more, for a 1-year survival rate of 100%. Four patients subsequently underwent surgical resection; complete necrosis of the tumor was observed in all four. The authors conclude that a combination of PEI and TAE is an appropriate treatment for patients with large, encapsulated HCC lesions who are poor surgical risks.

Aged↗

PubMed1991

Restoration of circadian corticosterone rhythm in ventromedial hypothalamic lesioned rats.

The effects of ventromedial hypothalamic (VMH) lesions on the circadian periodicity of blood corticosterone were studied in female rats. The rats were kept on a 12-hour light/12-hour dark illumination regimen and fed ad libitum. Three, 10 and 12 weeks after the VMH lesions, the concentrations of blood corticosterone were measured every 4 h for 48 h in the same unanesthetized rats. Three weeks after the operation, the circadian rhythm in VMH-lesioned rats was disturbed and disappeared. The corticosterone levels at 03:00, 07:00, and 11:00 were significantly higher than those in sham-operated rats. 10 and 12 weeks after the operation, the circadian rhythm, however, was notable (p less than 0.05, p less than 0.05). The elevated mean corticosterone levels over 48 h at 3 weeks after the operation decreased at 10 and 12 weeks. The sham-operated rats showed a significant circadian rhythm at 3, 10 and 12 weeks after the operation (p less than 0.001 in each period) with a peak concentration at 19:00 and through at 07:00. These findings show that the corticosterone circadian rhythms which were disturbed in the dynamic phase after VMH lesions recovered in the static phase of obesity, suggesting that the ventromedial hypothalamus is not an essential biological clock of circadian corticosterone rhythm.

Animals↗