Biomedical subjects
Publications and source records attributed to K Satoh.
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The lamprey (Entosphenus japonicus Martens) has been recommended in Japan as an efficacious diet for curing some of chronic inflammatory disorders. Fresh lamprey is rich in n-3 unsaturated fatty acids (e.g., eicosapentaenoic and docosahexaenoic acids) with minimal seasonal variation. The dietary effect of lamprey oil on acute myocardial ischemia was therefore investigated. Rats were fed a control diet or a diet supplemented with 5% safflower oil (SO) or 5% lamprey oil (LO) for 4 weeks. After coronary artery ligation, rats fed the LO diet demonstrated a higher 24-h survival rate and a reduced loss of creatine kinase activity from ischemic myocardium when compared with rats fed the control or SO diet. Moreover, the intracellular redistribution of lysosomal enzyme activity in the ischemic myocardium was significantly suppressed in the LO diet group. Analysis of the fatty acid composition of myocardial phospholipids in rats fed LO showed marked elevation of n-3 fatty acids, whereas arachidonic acid (n-6) content was significantly reduced. Hence, the n-3/n-6 ratio of myocardial phospholipids was markedly increased in the LO diet group. In addition, there was more than a 77% reduction in TXB2 synthetic capacity in whole blood in rats fed the LO diet when compared with rats fed either the control or SO diet. In conclusion, dietary supplementation with LO prevents sudden cardiac death and limits the extension of cellular damage from acute myocardial ischemia in rats. Both changes in thromboxane generation and altered membrane fatty acid composition may be involved in the observed reduction of ischemic damage in the heart.
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Recently percutaneous transluminal coronary recanalization therapy (PTCR) with urokinase infusion has became one of popular technique for coronary arterial occlusion. This paper reported clinical experience of intraarterial urokinase infusion therapy for acute or superacute stroke patients. The procedure was followed by angiographical study which revealed the major intracerebral arterial occlusion in three cases. Case 1: A 74-year-old female had sudden onset of clouding of consciousness with complete left hemiplegia. The patient was in our urological ward because of treatment for her right ureter tumor, as the patient was immediately subjected to angiographical study and complete occlusion of the trunk of the right middle cerebral artery was revealed four hours after onset. Successively 240,000 IU of urokinase solution was injected through the arterial catheter after angiographical study. This procedure repeated two times with 10 minute intervals. So total amount of 720,000 IU of urokinase was given by intraarterial injection. Immediately after the last urokinase injection the patient started to recover her consciousness and weakness. Simultaneous angiogram demonstrated partial recanalization of the proximal branches of the middle cerebral artery. The following day, she had complete recovery from her neurological deficits although she had transient hemorrhagic tendency. The final angiogram showed no existence of obstructed cerebral arteries as well as no low density areas in computed tomographic images. Case 2: A 73-year-old female, with the left internal carotid occlusion at the site of C1-2 portion, was instituted infusion therapy of similar procedure with total amount of 960,000 IU oi urokinase ten to twenty hours after onset. However, no rewarding was obtained.(ABSTRACT TRUNCATED AT 250 WORDS)
Concentrations of cefuroxime (CXM) in the skin and the serum of 24 patients were examined. CXM (0.75 g, 1.5 g) was administered to these patients intravenously in one shot. The skin and the serum were collected at 30 and 60 minutes after the injection. Concentrations of CXM in the skin were following: 0.75 g, 30 min.; 11.4 +/- 5.3 micrograms/g 60 min.; 9.5 +/- 4.7 micrograms/g 1.5 g, 30 min.; 17.1 +/- 6.8 micrograms/g 60 min.; 15.7 +/- 13.2 micrograms/g Concentration ratios (skin/serum) at 60 minutes after the injection were from 72.5% to 79.2%. The binding ratio of CXM to serum protein was 41%, which was lower than most other antibiotics. High concentration levels of CXM were maintained in the skin for a long time. Based on this study, we consider that CXM is effective in the skin surgery.
The relative incidence of endometrial carcinoma is obviously increasing as a disease of post-menopausal women with the ageing of the population in Japan. Growth of malignant endometrium tends to develop there both into the muscle and into the cavity. However, diagnosis in most cases is made by stage 1a (39.7%) and 1 b (19.0%) in FIGO classification. In the treatment of endometrial carcinoma, even after a simple hysterectomy in 89.2% of total cases 5-year cure rate may be reached to 80%, which has compared well with the 60% cure rate for cervical cancer. Some fractions of endometrial adenocarcinoma are estrogen-dependent and could therefore respond to antagonistic influences of progestogens. In a recent understanding, above theoretical grounds for hormone dependency and responsiveness of endometrial malignant lesions has been substantiated by data provided by steroid receptor determination. The high estrogen and progesterone receptor concentrations found in 70% of endometrial adenocarcinoma support the principle that endometrial adenocarcinoma is related to abnormal estrogenic stimulation. Those cancer are used as a model for in vitro and clinical evaluation of progestogen therapy.
We studied the effects of recombinant erythropoietin (r-Epo) and thrombocytopenic serum (TS) on the cytoplasmic maturation of megakaryocytes derived from colony-forming units megakaryocyte (CFU-M). Serotonin content, ATP content, acetylcholinesterase (Ach-E) activity per megakaryocyte, and electron microscopic analysis were selected as markers of cytoplasmic maturation. Megakaryocytes induced by pokeweed mitogen-stimulated spleen cell-conditioned medium (PWM-SCM) alone showed low levels of ATP content and Ach-E activity, which did not increase during culture, as well as a low level of serotonin content, which gradually accumulated during day 7 of culture. When r-EPo was added to the culture system on day 3 after megakaryocytic colony formation with PWM-SCM, the serotonin content in megakaryocytes increased markedly but ATP content and Ach-E activity did not increase significantly. In contrast, TS caused an increase in ATP content and Ach-E activity, but did not cause an increase in serotonin content. Electron microscopic analysis showed that the demarcation membrane system (DMS) developed defectively only in local areas with the addition of r-Epo and PWM-SCM, whereas the DMS developed normally and dense granules were generated to near normal with the addition of TS. Recombinant Epo may act on the early stage of cytoplasmic maturation, whereas TS may act on the late stage of cytoplasmic maturation. The results shown herein suggest that at least two different factors may be necessary for full in vitro cytoplasmic maturation of megakaryocytes derived from CFU-M.
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A new acidic form of glutathione S-transferase (GST, pI 6.2) was purified from rat brain by S-hexylglutathione affinity chromatography followed by chromatofocusing. This form occupied 20-25% of the total activity bound to the affinity column. It had a molecular mass (subunit 26 kDa) similar to that of a major GST form of rat testis (MT or 6-6) on sodium dodecyl sulfate/polyacrylamide gel electrophoresis. However, it differed from the MT in isoelectric point, activity towards 1,2-dichloro-4-nitrobenzene and immunological properties. On two-dimensional gel electrophoresis the brain form gave a spot which was identical in molecular mass, isoelectric point and immunological properties to a less acidic one (Yn1) of two spots (Yn1 and Yn2) of the testis GST-MT. Therefore, the brain acidic form is a homodimer, and named GST-Yn1Yn1. The activity was inhibited by sulfasalazine, an inhibitor of leukotriene-C4 synthase. This form (GST-Yn1Yn1) showed the highest leukotriene-C4 synthase activity, 496 nmol/mg protein in 5 min, among nine cytosolic GST isoenzymes from the rat. The Km values for leukotriene A4 and glutathione were 26 microM and 3.5 mM respectively. A major GST form of rat brain, occupying about 40% of the total activity, was identical with GST-P (7-7) purified from rat liver bearing preneoplastic hyperplastic nodules and localized at astroglias. GST-P also showed the significant leukotriene-C4 synthase activity, 67.2 nmol/mg protein in 5 min, but the Km for leukotriene A4 was 100 microM, fourfold higher than that of GST-Yn1 Yn1. These results suggest that mainly GST-Yn1 Yn1 may be involved in leukotriene-C4 synthesis in rat brain.
Cultures of mesothelial cells from bovine pericardium were established and their arachidonate metabolism was characterized. The identification of the cultured cells was based on morphological observations, and by electrophoretic analysis of cytoskeletal proteins, which demonstrated a pattern previously reported for mesothelial cells. Factor VIII-related antigen was present by indirect immunofluorescence, but the cells had no thrombomodulin activity. The cultured pericardial cells metabolized arachidonic acid to 6-ketoprostaglandin F1 alpha and a small amount of prostaglandin E2. The same metabolites were produced by pieces of intact parietal pericardium but not by pieces from which mesothelium had been removed. The cultured mesothelial cells produced 94.6 +/- 60.4 (mean +/- S.D.) ng/mg (n = 3) cell protein of 6-ketoprostaglandin F1 alpha in response to the calcium ionophore A23187, 117.3 +/- 13.6 ng/mg (n = 3) with exogenous arachidonic acid, 18.3 +/- 11.3 ng/mg (n = 5) with bradykinin, 8.4 +/- 4.3 ng/kg (n = 4) with histamine and 11.2 +/- 9.7 ng/mg (n = 5) with thrombin. All of these values were significantly higher (P less than 0.05) than the control (2.1 +/- 1 ng/mg; n = 5). From these results, we conclude that the mesothelial cells account for the arachidonate metabolism in the pericardium. The production of prostaglandin I2 occurs in response to physiological or pathological, agonists, and is substantial. That is, it is approximately the same as endothelial cells. The release of eicosanoids by mesothelial cells into the pericardial space may have a significant role in cardiac physiology and pathology.
An understanding of the organization of cholinergic neurons in the central nervous system has been an important objective for many years. By developing and applying a new electronic method for mapping tissue sections, we have generated original graphic and quantitative findings on forebrain cholinergic neurons that provide new insight into their distribution and organization. Satoh, Armstrong, and Fibiger (Brain Res. Bull. 11:693-720, 1983) have proposed that in the basal forebrain cholinergic neurons with long axons form a continuum rather than being arranged as a series of discrete nuclear groups. It has been difficult, however, by conventional methods of data analysis and display, to test this hypothesis. By using a digital microscopy system, the position of every cholinergic neuron was marked with 1-micron resolution in tissue sections taken at 90-microns or 180-microns intervals through the entire distribution of these neurons in the forebrain. The three-dimensional reconstruction of these neurons in context shows them to be distributed as a continuous cell column. The column twists and changes position as it is deformed by adjacent neuronal structures, such that its shape and continuity would not be apparent without reconstruction into a computer graphics model. Complementary analyses of the distribution of cholinergic interneurons in dopamine-rich regions of the forebrain indicated that there are regional differences between striatal and olfactory tubercle neurons. Cellular morphometry analyses show the population of cholinergic neurons in the rat to be surprisingly homogenous in size, but not in shape. Graphic and quantitative analyses indicated that there is a striking relationship between the distributions of projection and interneuronal cell groups. We conclude that the basal forebrain cholinergic neurons form a continuum. The chemoarchitecture of this cell group does not conform to the usual cytoarchitectural divisions. The present results, however, taken together with the findings based on Nissl-stained sections and connectional and biochemical data, suggest that the region of this neurochemically defined continuum should be reexamined for consideration as a single functional entity or nucleus: a cholinergic basal nuclear complex.
alpha 1-Antitrypsin (alpha 1AT) is a highly pleomorphic 52-kDa serum glycoprotein that functions as the major inhibitor of neutrophil elastase. Of these, the most common normal alpha 1AT haplotypes identified by isoelectric focusing (IEF) of serum are those of the M family, including M1, M2, and M3. In the course of studying the alpha 1AT type Z gene, we identified a restriction endonuclease BstEII polymorphism in the M1 gene that predicted the existence of a previously unidentified, but relatively common, haplotype of M, referred to as M1(Ala213) [Nukiwa, T., Satoh, K., Brantly, M. L., Ogushi, F., Fells, G. A., Courtney, M., & Crystal, R. G. (1986) J. Biol. Chem. 261, 15989-15994]. In this study we have cloned both alpha 1AT genes from an individual heterozygous for the M1(Ala213) and M1(Val213) haplotypes. Sequencing of the coding exons of both demonstrated that they are identical except for the Ala-Val difference at residue 213. The codominant transmission of the M1(Ala213) gene was demonstrated in a family study. Evaluation of 39 genomic samples of Caucasians with the IEF haplotype M1 demonstrated haplotype frequencies of 68% for M1(Val213) and 32% for M1(Ala213). alpha 1AT serum levels of individuals inheriting the M1(Ala213) gene in a homozygous fashion were in the same range as those for homozygous M1(Val213) as was the rate of association of the M1(Ala213) protein with neutrophil elastase.(ABSTRACT TRUNCATED AT 250 WORDS)
We evaluated the effects of dopamine (DA) and synthetic atrial natriuretic polypeptide (ANP) on the release of catecholamines (CA) from the adrenal medulla. Adrenal glands of male Wistar rats were superfused with Ringer's solution saturated with 95%, O2, 5% CO2 by the use of a continuous flow incubation system, and norepinephrine (NE) and epinephrine (E) concentrations in the perfusate were continuously measured by high pressure liquid chromatography with fluorescent reaction. And the effects of DA and ANP on the CA release were evaluated. Next the effects of metoclopramide (MC), dopamine (D2) antagonist, and glucagon were added in the Ringer's solution, and the changes of NE and E in the perfusate were determined. Basal secretion of NE and E were 0.02-0.04 ng/mg.wet weight/min and 0.05-0.1 ng/mg.wet weight/min, respectively. DA remarkably decreased both NE and E release, and the suppressive effect was dependent on DA concentration in the perfusate. MC clearly raised NE and E release as well as glucagon. The increasing effect of MC was perfectly suppressed by 10(-4) M of DA. But the effect of glucagon was not blocked by the same dose of DA. Alpha rANP (10(-5)M) slightly decreased the releases of NE and E from adrenal medulla, and the magnitude of the effect of rANP was smaller than that of DA. MC significantly increased NE and E release even when the adrenal gland was superfused with Ringer's solution containing 10(-5)M of rANP. These data suggest that the release of CA from adrenal medulla may be regulated by DA, and that the receptors specifically binding to DA may exist in adrenal medulla as well as sympathetic presynaps. We concluded that DA (but not ANP) may play an important role in controlling (suppressing) the activity of sympathoadrenomedullary system.
A simple and rapid method for studying optical anisotropic properties of liposomes was proposed. Intensities of transmitted light through one spherical liposome of dipalmitoylphosphatidylcholine placed between two polarizers were measured at various wavelengths by a microscopic spectrophotometer. Large increases in the intensities were observed at the prephase-transition temperature, which were caused by an increase in the birefringence of the multilayer of the liposome. The birefringence values obtained from the intensity data were about 0.020 below the pretransition temperature and 0.028 above that temperature. These values are in good agreement with the results reported for the plane samples in which lipid bilayers are stacked. The obtained values of the birefringence were far lower than the values estimated from polarizabilities. This lower birefringence is attributed to disordering of the tilt direction in the multilayer. The degree of order of the liposome multilayers calculated from the birefringence increased by 38% at the pretransition. This simple method is applicable to the study of the multilayer structure of liposomes in water.
17 alpha, 20 alpha-dihydroxy-4-pregnen-3-one (17 alpha, 20 alpha-P) is reportedly a hyper-tensinogenic substance in a model of ACTH induced hypertension in sheep (Scoggins et al). This steroid has not previously been measured in Japan. We have developed a new radioimmunoassay (RIA) system for 17 alpha, 20 alpha-P, and measured this steroid in peripheral blood samples from non-pregnant and pregnant women. In this clinical investigation concentrations of 17 alpha-hydroxyprogesterone (17 alpha-OHP) and progesterone (P) were also measured in the same clinical samples. Specific anti-serum against 17 alpha, 20 alpha-P-3-carboxymethyloxime-BSA was generated in rabbits. Cross-reactions with 5-pregnen-3 beta, 17 alpha 20 alpha-triol (17 alpha, 20 alpha-P5) and 17 alpha-OHP were 42 and 2.4%, respectively. Other steroids showed cross-reactivity of less than 1%. As an internal standard dexamethasone (500 ng/100 microliter) was added to the samples which were extracted twice with 6 volumes of dichloromethane. Separation of the plasma extract was performed by high performance liquid chromatography (HPLC) with the ODS column in the solvent system, acetonitrile:water = 55:45. Eluates of the 17 alpha, 20 alpha-P and 17 alpha-OHP fractions were collected separately and the solvent was evaporated in vacuo. RIA was carried out on each extract with specific anti-sera. RIA of P was performed independently using the anti-P-3-CMO-BSA antiserum; 0.1 ml of serum was extracted with n-hexane and subjected to RIA without chromatography. The coefficient of variation for intra- and interassay (n = 10) were 8.5 and 11.7% for 17 alpha, 20 alpha-P; 7.2 and 12.1% for 17-OHP and 10.5 and 14.0% for P, respectively. The recovery rates of 17 alpha, 20 alpha-P, 17 alpha-OHP and P were 95 +/- 1.5, 93.2 +/- 3.5 and 97.5 +/- 4.2%, respectively. With the use of this method, 17 alpha, 20 alpha-P was measured in the blood samples from the menstrual cycle and normal pregnancy. The mean serum 17 alpha, 20 alpha-P concentrations +/- SD in the follicular, ovulatory and luteal phases, and in the first, second and third trimesters were; 76 +/- 6.0, 100 +/- 31, 101 +/- 6.4, 226 +/- 80, 151 +/- 48 and 367 +/- 99 pg/ml, respectively.(ABSTRACT TRUNCATED AT 400 WORDS)
Evidence is presented that a distinctive type of glutathione-S-transferase (GSHTase-P), and a cytosolic polypeptide of Mr 52,000 (P-52), each appearing in greatly increased amounts in hepatocyte nodules during liver carcinogenesis in the rat, are so far indistinguishable. The probable identity of the two polypeptides was established with the use of SDS polyacrylamide gel electrophoresis and Western blot techniques with purified GSHTase-P and P-52 and their respective antibodies and by comparison of the sequence of the first 26 N-terminal amino acids. Since the enzyme and the polypeptide are each considered to be the best available early markers for hepatocyte nodules, as putative precancerous lesions, their probable identity makes them attractive cellular components for in depth studies on their transcriptional and translational regulation and their use in new approaches to the sequential analysis of liver carcinogenesis.