Biomedical subjects

S Fujimoto

Publications and source records attributed to S Fujimoto.

At least 541 records · Page 30Linked to original sources

PubMed1993

Vasocontractions of the in-vitro toad aortas induced by endothelin-1 and sarafotoxin-S6b.

Dose-dependent tension curves were recorded from the in vitro toad aortas by administration of endothelin-1 and sarafotoxin-S6b. The maximal contractile tensions by both drugs were evoked at a 10(-8) M concentration. By a single dose application (10(-8) M) of endothelin-1 and sarafotoxin-S6b to both endothelium-preserved and denuded vessels, the induction of the endothelium-dependent vasocontraction occurs after 2 min of administration. Ultrastructural changes of Weibel-Palade bodies such as decrease in electron density, swelling with a wide peripheral halo, and expulsion of their contents in a manner of exocytosis become evident within 2 min after administration of these drugs. These findings indicate that some vasocontractile substances in Weibel-Palade bodies are extracellularly discharged by endothelin-1 and sarafotoxin-S6b.

Animals↗

PubMed1993

Immunoelectron microscopy on the localization of endothelin in the umbilical vein of perinatal rabbits.

Immunoelectron microscopic localization of endothelin (ET) using the mouse monoclonal anti-ET sera was performed on the pre- and postnatal rabbit umbilical vein during stages when Weibel-Palade (WP) bodies increase in number in the endothelial cells. The immunoreactions are localized in the rough endoplasmic reticulum, Golgi cisterns, and WP bodies that are actively segregated from the Golgi apparatus. After degranulation and extracellular release of WP bodies was induced by compound 48/80, heavy immunoreactions were seen in both vascular lumen and cytoplasm near the degranulated WP bodies. These results indicate that ET is stored in WP bodies after segregation from the Golgi apparatus and released from the endothelial cells concomitantly with other components of these granules by exocytosis. Because the immunoreactions are also seen in pinocytotic vesicles that are occasionally in contact with WP bodies, it seems likely that WP bodies are also involved in the uptake and storage of ET from blood plasma.

Animals↗

PubMed1993

Effects of coenzyme Q10 administration on pulmonary function and exercise performance in patients with chronic lung diseases.

Serum coenzyme Q10 (CoQ10) levels were measured at rest and during incremental exercise in 21 patients with chronic obstructive pulmonary disease (COPD) and 9 patients with idiopathic pulmonary fibrosis (IPF). The mean serum CoQ10 levels at rest in patients with COPD and IPF were 0.56 +/- 0.20 and 0.45 +/- 0.16 microgram/ml, respectively. In both groups these levels were decreased compared with those of healthy subjects. In the patients with COPD, CoQ10 levels were significantly correlated with body weight, however, there was no correlation between CoQ10 levels and ventilatory function, PaO2, VO2/kg at rest, or maximal VO2. In eight of nine patients whose PaO2 at rest was lower than 75 torr, serum CoQ10 levels were lower than 0.5 microgram/ml. We studied the effects of the oral administration of CoQ10 at 90 mg/day for 8 weeks on pulmonary function and exercise performance in eight patients with COPD. Serum CoQ10 levels were significantly elevated in association with an improvement in hypoxemia at rest, whereas pulmonary function was unaltered. Oxygen consumption during exercise was not changed, whereas PaO2 was significantly improved, and heart rate was significantly decreased compared with the results obtained at an identical workload at baseline. Furthermore, lactate production was suppressed during the anaerobic exercise stage after CoQ10 administration, and exercise performance tended to increase. These data suggested that CoQ10 has favorable effects on muscular energy metabolism in patients with chronic lung diseases who have hypoxemia at rest and/or during exercise.

Aged↗

PubMed1993

Combined treatment of pelvic exenterative surgery and intra-operative pelvic hyperthermochemotherapy for locally advanced rectosigmoid cancer: report of a case.

A huge rectosigmoidal cancer which extended into the urinary bladder in a 64-year-old man is herein described. The tumor occupied the pelvic and lower abdominal cavities, while the rectosigmoid was totally obstructed. No hepatic or pulmonary metastasis was evident. The ventral and flank sides of the peritoneum in the right lower abdomen, right common iliac vessels, bilateral ureters, terminal ileum, cecum, ascending colon, and urinary bladder were all directly invaded by the tumor, but the aorta, sacrum, and lower rectum were free of cancer. Consequently, an anterior pelvic exenteration was carried out along with an ileal conduit and a right hemicolectomy. Immediately after the exenteration, intra-pelvic hyperthermochemotherapy was performed using a 46-47 degrees C perfusate containing 40 micrograms/ml of mitomycin C (MMC) and 200 micrograms/ml of cisplatin (CDDP), for 90 min, in an attempt to prevent any further local recurrence. A right hemicolectomy and a permanent colostomy were done simultaneously with the hyperthermia treatment. After an uneventful postoperative course, the patient was prescribed adjuvant chemotherapy, i.e., two administrations of 17 mg/m2 and 21 mg/m2 of MMC, and ten doses of 710 mg/m2 of 5-fluorouracil (5-FU) followed by five doses of 535 mg/m2 of 5-FU. At the time of this writing, the patient is still alive without recurrence at 21 months after surgery.

Adenocarcinoma↗

PubMed1993

Metabolic changes in cimetidine treatment for scald injury on the peritoneo-serosal surface in far-advanced gastric cancer patients treated by intraperitoneal hyperthermic perfusion.

Since pretreatment with cimetidine results in the prevention of scald injury on the peritoneo-serosal surface caused by intraperitoneal hyperthermic perfusion (IPHP) for advanced gastric cancer, the diverse influence of IPHP on patients who were either given or not given cimetidine was studied both during and after IPHP treatment. Cimetidine 50 mg/kg was injected intravenously into 12 patients immediately prior to IPHP. There were no statistical background differences between the cimetidine and control groups (those not given cimetidine). The inflow and outflow temperatures of the hyperthermic perfusate in the control and cimetidine groups were 46.1 +/- 0.1 degree C and 44.1 +/- 0.1 degree C and 46.3 +/- 0.1 degree C and 44.2 +/- 0.04 degree C, respectively. Either the pre-IPHP hypothermia or IPHP in the control group resulted in a considerable increase in serum noradrenaline and adrenaline. The intravenous administration of cimetidine led to a stransient but moderate drop in the mean blood pressure as well as a delayed appearance of high concentrations of noradrenaline and adrenaline, induced by high concentrations of circulating histamine released with cimetidine. These results suggest that the sympathetic nervous responses were activated either by hypothermia or hyperthermia. The transient hypotension and delayed increases of both serum catecholamines were attributed to a marked increase in circulating histamine, released with the intravenous cimetidine.

Blood Pressure↗

PubMed1993

Steroid sulfatase deficiency in Japanese patients: characterization of X-linked ichthyosis by using polymerase chain reaction.

X-Linked ichthyosis (XLI) due to deficiency of steroid sulfatase (STS) of which gene consists of 10 exons is an inherited skin disorder. The gene, mRNA and protein of STS were examined in six Japanese patients with XLI. Neither the mRNA nor the enzyme protein was detected in a patient. The results of Southern analysis using STS cDNA as a probe indicated that all the patients examined exhibited large deletions of the STS gene. When exon 1 and the exon 10 of the STS gene were amplified by polymerase chain reaction using patients' genomic DNA as templates, no product was detected in all the patients examined. These observations suggest that most XLI in Japanese patients is caused by an extensive deletion of the STS gene as was demonstrated in Caucasian patients. The PCR method in the present study is useful for the diagnosis of XLI in prenatal and postnatal subjects.

Arylsulfatases↗

PubMed1993

Morphological characterization of a new human epithelioid sarcoma cell line, ES020488, in vitro and in vivo.

A new human epithelioid sarcoma cell line (ES020488) was established from a cutaneous metastasis in 26-year-old man, and was morphologically characterized in vitro and in vivo by comparison with the original tumor. The ES020488 cells showed a male karyotype ranging from 39 to 83 chromosomes, with various abnormalities but no specific pattern. The cells were round, polygonal or spindle-shaped with abundant cytoplasm and round nuclei containing prominent nucleoli; they proliferated in a sheet-like pattern. Tumors transplanted into nude mice revealed essentially the same features as the original tumor. Both in vitro and in vivo, the cells immunohistochemically expressed vimentin, cytokeratin, and EMA, but not desmin and S-100 protein. Ultrastructural study revealed irregular or round nuclei containing abundant euchromatin and prominent nucleoli, many intermediate filaments running irregularly or around the nucleus, and a number of filopodia-like processes. ES020488 cells were thus proven to retain and exhibit the unique morphological characteristics of an epithelioid sarcoma both in vitro and in vivo. These cells are possibly derived from synovioblastic mesenchyme.

Adult↗

PubMed1993

Quantitative analysis of smooth endoplasmic reticulum proliferation in periportal, midzonal and perivenular hepatocytes of mice after administration of phenobarbital.

Smooth endoplasmic reticulum (SER) proliferation after phenobarbital (PB) administration has been described to occur predominantly in perivenular hepatocytes. We analyzed by quantitative electron microscopy changes in ER amounts in periportal, midzonal and perivenular hepatocytes from mice injected with 35, 50, 100 or 150 mg/kg of PB once a day for 3 days. The SER proliferated in midzonal hepatocytes in addition to perivenular hepatocytes even when low doses (35 or 50 mg/kg) were administered, and in hepatocytes of all three zones in case of high doses (100 or 150 mg/kg). Moreover, net proliferation of SER after administration of an amount of PB was not different in hepatocytes of any zones where the proliferation occurred, except animals injected with 100 mg/kg. The results suggest that there is no difference in SER producing capacity in hepatocytes of three zones when the cells have recognized the stimulation of PB.

Animals↗

PubMed1993

Membrane currents of porcine granulosa cells in primary culture: characterization and effects of luteinizing hormone.

In ovarian granulosa cells, LH depolarizes the membrane potential and induces steroid production. The membrane currents of porcine granulosa cells in primary culture were studied by means of the whole cell configuration of the patch-clamp technique to investigate whether the ionic channels are involved in the effects of LH. We identified and characterized two types of K+ currents--a transient outward current (Ito) and a delayed rectifier K+ current (IK)--and one Ca2+ current (ICa). Ito and IK were voltage- and calcium-dependent. Both currents were blocked by 4-aminopyridine, a K+ channel blocker, but only IK was sensitive to tetraethylammonium, another K+ channel blocker. ICa was inactivated within 50 ms of the test pulse. Nifedipine and verapamil, L-type CA2+ channel blockers, did not suppress ICa even at a concentration of 10 microM. Tetramethrin (1 microM), a T-type Ca2+ channel blocker, decreased ICa by 38.4 +/- 5.6% (n = 3). These findings suggest that the current may be a T-type Ca2+ current. LH and dibutyryl cAMP, potent stimulants of steroid production, attenuated Ito only by 13.9 +/- 1.8% (n = 7) and 21.0 +/- 1.5% (n = 4), respectively. However, they did not affect IK and ICa. These results indicated that LH does not modulate CA2+ current directly, but it slightly decreased Ito through cAMP elevation. The LH-induced inhibition of Ito may be involved in the depolarizing effect of LH and may play some role in steroid secretion or other functions in granulosa cells.

4-Aminopyridine↗

PubMed1993

Heterogeneity of HLA-G genes identified by polymerase chain reaction/single strand conformational polymorphism (PCR/SSCP).

A genomic HLA-G clone named 7.0E was isolated from a Japanese placenta. The deduced amino acid sequence of the 7.0E was identical to two HLA-G genomic clones and two cDNA clones previously described. The DNA sequences of alpha 1 and alpha 2 domains of the HLA-G gene from 5 cell lines also encoded the same amino acids. However, a 14 bp insertion, ATTTGTTCATGCCT, was present in the 3' untranslated region of 7.0E compared with the originally described HLA-G clone (HLA 6.0). Polymerase chain reaction (PCR)/single strand conformational polymorphism (SSCP) analysis of exon 8 allowed the HLA-G gene to be classified into two alternative types, G6.0 and 7.0 E, those correlated to the absence or the presence of the 14 bp stretch. Each group had minor sequence variant(s), and the alleles of the 7.0E-type were more heterogeneous than those of the G6.0-type. The 14 bp deletion is present only in the G6.0-type of HLA-G alleles among HLA class I genes. Thus it was suggested that G6.0 alleles were generated after diversification of the HLA-G.

Alleles↗

PubMed1993

Establishment of a feline T-lymphoblastoid cell line susceptible to feline immunodeficiency viruses.

A feline T-lymphoblastoid cell line susceptible to feline immunodeficiency viruses (FIV) was established and designated as Yu-1 cells. Yu-1 cells continued to grow over one year with more than 100 successive passages in the presence of human recombinant interleukin-2. Surface antigens of Yu-1 cells were feline Pan-T+, CD4+, and CD8-. Susceptibility of Yu-1 cells to FIV strains were higher than that of the primary culture of the feline peripheral blood mononuclear cells, indicating that this cell line should be useful for isolation, propagation, and neutralization test of FIV.

Animals↗

PubMed1993

T cell response to embryonal carcinoma F9 cells: induction and characterization of T cell receptor alpha beta+ double-negative cytotoxic T lymphocytes.

We investigated the mechanism of T cell response to murine embryonal carcinoma F9 cells. Thy-1+, CD4-, CD8- (double-negative) cytotoxic effector cells were induced in spleen cells obtained from immune A.BY mice to F9 cells, and the cytotoxic activity was major histocompatibility complex (MHC)-unrestricted. Furthermore, CD4+ T cells were essential for the induction of double-negative cytotoxic T lymphocytes directed to F9 cells. Most of the double-negative cytotoxic T lymphocyte lines obtained by long-term culture of the effector cells had CD3 molecule and T-cell receptor beta chain on their cell surface, and the CD3 molecule was found to be involved in target cell recognition. The T cell receptor alpha beta+ double-negative cytotoxic T lymphocyte line (2A5) also lysed various tumor cells in a non-MHC-restricted manner, but did not lyse concanavalin A-stimulated blasts of 129 strain, from which F9 cells had originated. These results indicate that T cell receptor alpha beta+ double-negative cytotoxic T lymphocytes induced by F9 cells recognize a common antigen(s) expressed on F9 cells and other tumor cells but not minor histocompatibility antigens.

Animals↗

PubMed1993

Difference between renal failure associated with methylprednisolone pulse therapy and deterioration of renal function unrelated to methylprednisolone therapy.

In this study, we attempted to analyze the differences between renal failure associated with methylprednisolone (MP) pulse therapy and natural deterioration of renal function that was unrelated to MP administration. Of 80 patients with renal or collagen disease who received MP pulse therapy at our hospitals, 13 were selected for the study whose serum creatinine levels increased more than 0.5 mg/dl from baseline values following therapy. Somewhat arbitrarily, 7 patients were placed in an MP-associated renal deterioration group (group 1) in which serum creatinine levels returned naturally, or following induced diuresis, to baseline levels, and 6 patients in an MP-independent natural deterioration group (group 2) in which renal function progressively deteriorated. Renal function similarly deteriorated in the two groups following pulse therapy, irrespective of the degree of crescent formation. Our data suggested that hypoproteinemia is the most important index for differentiating MP-associated renal failure from natural deterioration of renal function unrelated to MP pulse therapy. In patients that are nephrotic and have impaired renal function, worsening of renal function following pulse therapy may partly be due to transient MP-associated renal failure. On the other hand, in patients without hypoproteinemia, worsening of renal function is most likely due to active primary disease and is probably not associated with MP pulse therapy.

Aged↗

PubMed1993

Doppler echocardiographic assessment of left ventricular diastolic function in patients with progressive systemic sclerosis.

To investigate the diastolic function of the left ventricle in patients with progressive systemic sclerosis (PSS), 13 patients with PSS (3 males and 10 females; group PSS) were examined by Doppler echocardiography. As a control 8 age-matched healthy volunteers (3 males and 5 females; group H) were also examined. Pulse-wave transmitral Doppler flow velocity curves were digitized and curves of the first derivatives were obtained. In group PSS, total left ventricular wall thickness and left atrial dimension were increased, the ratio between the left ventricular end-diastolic dimension and the left ventricular wall thickness was decreased, late peak diastolic filling was increased, isovolumic relaxation time was prolonged, compared with group H. The patients with left ventricular wall thickening out of proportion to left ventricular end-diastolic dimension had decreased early peak diastolic filling. We conclude that patients with PSS have deterioration of left ventricular diastolic function, and the diastolic function is severely disturbed in patients with left ventricular wall thickening out of proportion to left ventricular end-diastolic dimensions.

Diastole↗

PubMed1993

Densities of NADPH-ferrihemoprotein reductase and cytochrome P-450 molecules in the endoplasmic reticulum membrane of rat hepatocytes.

In hepatocytes, NADPH-ferrihemoprotein reductase (reductase) has been hypothesized to exist as aggregates or micelles in endoplasmic reticulum (ER) membrane. However, if the number of reductase molecules per unit area of ER is low, this hypothesis cannot explain how a few reductase molecules efficiently reduce many P-450 molecules. To test this hypothesis, we estimated the numbers of reductase and P-450 molecules per unit ER area (reductase and P-450 densities) by microphotometry of the two enzymes in conjunction with morphometry of ER in periportal, midzonal, and perivenular rat hepatocytes. The reductase density in periportal, midzonal, and perivenular hepatocytes (107-179 molecules/microns 2 of ER) was high enough to efficiently reduce all P-450 molecules in the ER, although the value in perivenular hepatocytes was lowest owing to the relatively greater amount of ER in this region. The pattern of sublobular gradient in the reductase density was similar to that in the P-450 density. Consequently, the molar ratio of P-450 to reductase in ER was similar (about 40:1) in hepatocytes regardless of their positions within the liver lobule.

Animals↗

PubMed1993

Site-specific inactivation of papain by ascorbic acid in the presence of cupric ions.

The mechanism of inactivation of papain (EC 3.4.22.2) by ascorbic acid (AsA) in the presence of cupric ions (Cu2+) was investigated. The aerobic combination of Cu2+ and AsA resulted in an irreversible loss of enzyme activity. The inactivation was found to be an apparent first order reaction. The prior mixing of Cu2+ and AsA caused the complete disappearance of the inactivation. The addition of iron ions led to significant suppression against the inactivation. Cu2+ was bound to the enzyme in a molar ratio of 1:1. At lower concentrations of Cu2+ (molar ratio of enzyme to Cu2+ of 1: < 1), the extent of inactivation showed the same dependence against the extent of oxidation of AsA. The rate of inactivation increased as the concentration of AsA was increased. Saturation kinetics were observed with respect to the concentration of AsA. Changes in the concentration of Cu2+ had no effect on the dissociation constant of the enzyme-AsA complex (KI), though the rate constant of inactivation (k2) showed a linear relationship with the concentration of Cu2+. At various pH values tested, no change of k2 was found, whereas the value of KI increased when the pH bacame lower. At higher concentrations of Cu2+, the rate of inactivation fell beyond a certain concentration of AsA. The present results suggest that both Cu2+ and AsA bind to the enzyme to form a ternary complex and that free radicals are site-specifically formed and react preferentially with the enzyme, at the site of their formation, impairing its activity.

Ascorbic Acid↗

PubMed1993

Formation of a hydroxyl radical by the myeloperoxidase-NADH-oxygen system.

When phenylalanine was incubated with myeloperoxidase (MPO) and NADH in citrate buffer (pH 4.5), o-, m-, and p-tyrosines were identified as hydroxylated products. Tyrosine formation was dependent on the reaction time and MPO concentration. No significant quantities of tyrosines were formed on if MPO was omitted and inactivated MPO was added instead of active MPO. The tyrosine formation by the MPO-NADH system was greatly reduced under anaerobic conditions, and significantly inhibited by hydroxyl radical scavengers. Superoxide dismutase was a potent inhibitor, but catalase was less effective. Even though the superoxide radical (O2-)-producing ability of the MPO-NADH system was about 29% of that of the hypoxanthine-xanthine oxidase system, under the experimental conditions employed, the rate of tyrosine formation from phenylalanine by two systems was found to be a similar. The above results suggest that the formation of a hydroxyl radical (OH.) may occur in the MPO-NADH system under aerobic conditions and a superoxide radical may be involved in the OH. formation, with MPO promoting the OH. formation from O2-.

Aerobiosis↗

PubMed1993

Purification and characterization of zinc-dependent acid phosphatase from bovine brain.

Zn(2+)-Dependent acid phosphatase (Zn(2+)-APase) was purified to homogeneity from bovine brain. The apparent molecular weight of the enzyme was estimated to be about 62000 by gel filtration and 31000 by polyacrylamide gel electrophoresis in the presence of sodium dodecyl sulfate. The enzyme exhibited an isoelectric point of approximately 4.8. The enzyme required Zn2+ ions for catalytic activity, but other cations had little or no effect. The maximum enzyme activity was obtained in the presence of about 5 mM of Zn2+ at pH 5.5 in 50 mM 2-(N-morpholino)ethanesulfonic acid-NaOH buffer. The enzyme significantly catalyzed the hydrolysis of p-nitrophenyl phosphate, phenyl phosphate, and phosphotyrosine. The enzyme was also active for myo-inositol-2-monophosphate and adenosine 2'-monophosphate of the other common phosphate esters tested, though significantly less active than for p-nitrophenyl phosphate. The optimum activity pH of the enzyme was around 5.5 with p-nitrophenyl phosphate and myo-inositol-2-monophosphate. The enzyme was resistant to fluoride ions. Two types of Zn(2+)-APases, a high molecular weight (molecular weight, M(r)., about 110,000) and a low molecular weight (M(r), about 62,000) type, were found to exist in various tissues of rat. Brain, lung, spleen, stomach, heart, skeletal muscle, and erythrocytes contained only the lower molecular weight type. On the other hand, liver and kidney contained mainly the higher molecular weight type, and the small intestine contained significant quantities of the both types.

Acid Phosphatase↗