Purification of growth factors from cartilage.
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Biomedical subjects
Publications and source records attributed to M Iwamoto.
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This study shows that rat thyroid follicular (FRTL5) cells can produce interleukin 6 (IL-6) spontaneously and that IL-1 beta enhanced the production of IL-6 dose-dependently. In addition, 10 ng/mL of IL-1 beta in combination with 10 to 1,000 microU/mL of thyroid-stimulating hormone (TSH) synergistically enhanced IL-6 production. It is suggested that IL-6 released from thyroid follicular cells may be a costimulator of autoreactive B and T cells in autoimmune thyroid disease and may constitute another link between the immune and endocrine systems.
We have previously suggested the existence of two distinctive states of cholesterol in erythrocyte and lymphoma cell membranes as revealed by high- and low-affinity binding sites for theta-toxin of Clostridium perfringens [Ohno-Iwashita, Y., Iwamoto, M., Mitsui, K., Ando, S., & Nagai, Y. (1988) Eur. J. Biochem. 176, 95-101; Ohno-Iwashita, Y., Iwamoto, M., Ando, S., Mitsui, K., & Iwashita, S. (1990) Biochim. Biophys. Acta 1023, 441-448]. To understand factor(s) which determine membrane cholesterol heterogeneity, we analyzed toxin binding to large unilamellar liposomes composed of cholesterol and phospholipids (phosphatidylcholine/phosphatidylglycerol = 82:18, mol/mol). Liposomes containing phospholipids with 18-carbon hydrocarbon chains at both positions 1 and 2 of the glycerol have both high- and low-affinity toxin-binding sites with Kd values similar to those of intact erythrocytes, whereas liposomes with hydrocarbon chains containing 16 or fewer carbons at either position 1 or 2 have only low-affinity toxin-binding sites. The cholesterol/phospholipid ratio, in addition to the length of phospholipid hydrocarbon chain, also determines the number of toxin-binding sites, indicating that at least these two factors determine the topology of membrane cholesterol by creating distinctively different affinity sites for the toxin. Since theta-toxin binding detects specific populations of membrane cholesterol that are not detectable by the measurements of susceptibility to cholesterol oxidase and cholesterol desorption from membranes, the toxin could provide a unique probe for studying the organization of cholesterol in membranes.
A highly specific competitive enzyme-linked immunosorbent assay for the epimastigote of Tulahuen strain was developed by using the usual 3 immunological reagents, a rabbit antiserum specific for T. cruzi, epimastigote of Tulahuen strain, beta-D-galactosidase-labeled goat anti-rabbit immunoglobulin G and the solid-phase cell fragments of the epimastigote of Tulahuen strain. A new method, the selected antibody enzyme immunoassay (SAEIA) which generally detected all strains of the epimastigote tested with the same working range, was developed by changing only the solid-phase antigen to the epimastigote of Y strain among the 3 immunological reagents. Both assays permitted us to measure accurately as little as 1,000 parasites per assay tube. Scope of the SAEIA was limited to the epimastigote. Both life-cycle forms of T. cruzi which appear in mammals, amastigote and trypomastigote, and other kinetoplastids showed low cross-reaction values by the assay. The assay principle of the new method and a preliminary study to apply the SAEIA for finding the field T. cruzi-infected insect vectors were also reported.
New 5'-nucleotidase inhibitors named NF-86I, NF-86II were recently isolated from the seeds of Areca catechu L. NF-86I and NF86II showed inhibitory effects on the growth of Streptococcus mutans MT8148(c) and Streptococcus mutans MT6715(g), respectively. In addition, these inhibitors could inhibit insoluble glucan formation from sucrose. NF-86I and NF-86II were found to be polyphenolic substances. Some polyphenols such as tannic acid bind non-specifically to proteins (tannic activity). The 5'-nucleotidase inhibitors that we isolated did not show any such activity. However, the growth inhibitory activity and the inhibitory effect on water-insoluble glucan production were equal to tannic acid. It is therefore considered that these inhibitors bind specifically to the bacterial cell surface. Our findings suggest that the 5'-nucleotidase inhibitors NF-86I and NF-86II may be useful anti-plaque preventing agents.
New 5'-nucleotidase inhibitors designated as NPF-88BU-IA, NPF-88BU-IB, NPF-88BU-IIA and NPF-88BU-IIB, respectively, were isolated from the seeds and skin of the wine grape "Koshu". They were purified by solvent extraction, dialysis, and reversed-phase high performance liquid chromatography (HPLC). Their physico-chemical properties revealed these compounds to be polyphenolic substances. The average relative molecular masses of the four were estimated by gel permeation chromatography (GPC) analysis to be 7850, 5950, 11900, and 11300, respectively. They strongly inhibited 5'-nucleotidase activities from snake venom and rat liver membrane, and displayed significant therapeutic activity against Ehrlich ascites carcinoma. They also showed inhibitory effects on the growth of Streptococcus mutans MT8148(c), a primary cariogenic bacterium. Furthermore, these 5'-nucleotidase inhibitors inhibited the glucan formation from sucrose. These results suggest that the 5'-nucleotidase inhibitors can prevent the cause of caries of tooth.
It has been reported that an acetone extract of ginger and its fractions have anti-5-HT (5-hydroxytryptamine; serotonin) effects. In the present study, guinea pig ileum, rat stomach fundus and rabbit aortic strips are used in order to determine the constituents of fraction 2 which are responsible for anti-5-HT effect and to examine their pharmacological properties. The analysis of fraction 2-3 indicated that galanolactone, a diterpenoid, is one of the active constituents. In guinea pig ileum, galanolactone inhibited contractile responses to 5-HT with a pIC50 value 4.93. pIC50 value of galanolactone against the response to 2-methyl-5-HT, a selective 5-HT3 agonist, in the presence of methysergide at 1 x 10(-5) M was 5.10. pIC50 values of ICS 205-930, a selective 5-HT3 antagonist, were 5.30 and 7.49, respectively. The concentration-response curve of 5-HT was shown as a biphasic curve and galanolactone caused a selective shift to the right of the second phase. In the same preparations, the pIC50 value of galanolactone and ICS 205-930 against the response to carbamylcholine (CCh) was 4.45 and 4.46. The inhibitory effect of galanolactone on the 5-HT response in the stomach fundus and aortic strips was less than that in the ileum. In addition, in the thoracic aorta precontracted with 50 mM K+, the relaxing effect of galanolactone was about 1/10 of that of papaverine. These results suggest that the anti-5-HT effect of galanolactone, a diterpenoid isolated from ginger, is related to antagonism of 5-HT3 receptors.
N-Terminal tripeptide analogs of fibrin alpha-chain were synthesized and their inhibitory effect on fibrinogen/thrombin clotting was examined. A new water-soluble active ester, 3-pyridinium ester, was used for the synthesis. Among the synthetic peptides, H-Gly-Pro-Arg-hexamethyleneimine exhibited the highest inhibitory effect on fibrinogen-thrombin clotting.
The preganglionic trunk of the superior cervical ganglion was implanted chronically into the carotid artery. A helical strip of the carotid artery with nerve was prepared, and electrical stimulation of the nerve caused a definite contraction. Part of the contraction was attenuated by phentolamine, and the remaining part of the contraction was nearly abolished by tubocurarine but not by hexamethonium and atropine. The present results clearly demonstrate that the carotid artery is reinnervated by preganglionic cholinergic fibers.
The effect of acupuncture points stimulation on the induction of plaque-forming cells (PFC) in spleen cells of BALB/c mice was investigated in vivo and in vitro tests. In in vivo experiment, mice were immunized with 2 x 10(8) sheep red blood cells (SRBC) and the PFC was markedly increased by daily (once a day for 4 days) acupuncture stimulation. The enhancement of PFC by acupuncture was completely blocked by preadministration of procain, hexamethonium, naloxone, propranolol, but not by phentolamine. The enhancement of PFC by acupuncture was also observed in spleen cells of non-immunoized mice when spleen cells of the acupunctured mice were cultured with SRB in vitro. The enhancement of PFC in spleen was observed after stimulation with acupuncture, and a similar effect was also found in bone marrow cells of normal mice, but not in thymic cells. The spleen cells of mice given acupuncture showed no enhancement of PFC after treatment with anti-Thy 1.2 antibody and complement. Furthermore, these helper T cells were found to be not restricted by the H-2 gene complex. These data demonstrate that the helper T cells induced by acupuncture lack the H-2 restriction, and thus suggest that they may be drived from the bone marrow, but not from the thymus. It is therefore concluded that the helper T cells derived from the bone marrow were activated via the sympathetic nervous system stimulated by acupuncture.
The aim of this study was to determine the effects of olfactory stimulation on immunological and behavioral states in mice. Anti-SRBC (IgM) plaque forming cell (PFC) count and spontaneous running activity (SRA) were measured to demonstrate the effects of exposure to a given fragrance. The decreased PFC count and thymic involution induced by high pressure stress in mice were recovered after exposing the stressed mice to the fragrance continuously for 4 days after the stress was given. The PFC and SRA also appeared to be maintained at normal levels by oflactory stimulation with the fragrance for 24 hrs after the given stress. The immunological suppression induced by high pressure stress was considered to be caused by the induction and activation of suppressor cells. However, exposure to the fragrance after the stress did not enhance suppressor activity. The restoration of the stress-induced immune suppression by olfactory stimulation was blocked by procain administration onto the olfactory cells.
Strategic recanalization of the occluded cerebral vessels has shown promise as a therapy for embolism and thrombosis in the acute stage. A single-dose of urokinase (UK) administered by intravenous and intraarterial routes was usually designed to restore patency of the infarct-related arteries and reperfuse the area of infarction. However, thrombolytic agents which have been available to date may lack resoluvability, limiting the amount of doses, because overdosage may induce hemorrhagic complication. This newly-designed therapy, intraarterial injection of UK-low molecular dextran (LMWD) complex was introduced in order to overcome the danger shown in the previous study. A high-resolvent allows cut-down of urkinase doses. Patients with acute cerebral infarction were selected for treatment with the resolvent if they satisfied the following conditions: 1) up to 79 years old without serious systemic diseases, 2) less than 12 hours from the onset, 3) better than the score of 8 in GCS, 4) no abnormality in CT scan, 5) apparent neurological deficit and 6) responsible pathology in angiography. LMWD of 15ml, UK of 240000IU and 15ml saline-complex was injected as one course at 2.0-2.5ml/min for 11 cases. Recanalization was observed in seven cases of embolism, and lack of reperfusion in five cases of thrombosis. The minimum effective dose was determined as 480000IU of UK in two courses. In terms of time lag from the onset, six hours may be the inferior limit, within which five of six cases (83%) succeeded in the recanalization.(ABSTRACT TRUNCATED AT 250 WORDS)
We examined 24 pediatric patients to evaluate the usefulness of sudden near maximal exercise test (dash method), where the subjects began to run at Bruce protocol of the last stage. 1) No considerable differences between two protocols were found in maximal oxygen consumption (V O2max), maximal heart rate (HRmax), maximal systolic blood pressure, and findings of electrocardiography. 2) The sudden maximal exercise test could be completed during a shorter period compared to Bruce protocol. 3) The sudden maximal exercise protocol reached 84% of HRmax and 47% of V O2max at one minute after the onset of the protocol, and produced 96% of HRmax and 89% of V O2max at two minutes after the onset. We considered that sudden maximal exercise protocol was useful to obtain a response similar to Bruce protocol at maximal exercise within a short period. We have to pay attention to the safety of the patients because their cardiopulmonary response to sudden maximal exercise protocol is dramatic.
A 30-year-old male died suddenly after two years of clinically progressive episodes of recurrent ventricular tachycardia. In spite of recurrent episodes of palpitation, the patient had never shown signs or symptoms of heart failure before death. Post mortem examination revealed massive left ventricular involvement in addition to complete absence of myocardial tissue in the right ventricle, replaced by adipose tissue. This case illustrates that patients with arrhythmogenic right ventricular dysplasia may die of arrhythmia before failure of the left ventricle.
Biophysical profile (BPP) score was assessed immediately before fetal blood sampling by cordocentesis in 150 fetuses referred to our hospital, 95 after and 55 before 30 weeks of gestation. In 95 fetuses after 30 weeks of gestation, 39 fetuses were evaluated with BPP scores of 12, 35 were from 8 to 11 and 21 were less than 7. In 55 fetuses before 30 weeks of gestation, 8 fetuses were evaluated with BPP scores of 12, 27 were from 8 to 11 and 20 were less than 7. pO2, pH and pCO2 in fetuses with a score less than 7, either before or after 30 weeks of gestation, (with a score less than 7) did not significantly differ, in comparison to the other two groups. No variables in the biophysical profile precisely reflect fetal hypoxemia, acidemia or hypercarbia. Since even the fetus with a BPP score of greater than 8 may not always be assured of well-being and not all fetuses with a score of less than 7 are necessarily in a deteriorated condition, it is necessary to evaluate fetal condition on the basis of fetal blood gas data obtained by cordocentesis, especially when the fetus is additionally handicapped by prematurity or morbidities such as growth-retardation.
We developed a method for a determination of the amount of retrogradely transported HRP in the rat masseteric motoneuron using a modification of Mesulam's HRP histochemical protocol and an image processing system combined with a light microscope and a television camera. To test the validity and reproducibility of the new method, a quantitative comparison of the amount of dark blue granules of HRP-product in the cell body of masseteric motor neurons was performed between the right and left trigeminal motor nuclei of 70 rats, which resulted in no significant difference. An additional study used the method was made of the effects of administration of five dopamine receptor antagonists with different biochemical and pharmacologic properties on retrograde transport of HRP in the rat masseteric motoneuron. As a result, chlorpromazine, haloperidol, SCH 23390, and sulpiride significantly enhanced retrograde transport of HRP as against domperidone which showed no significant change in the transport. A possible regulatory system for retrograde transport of HRP in the masseteric motoneuron was discussed in relation to the action of the dopamine receptor.
One thousand forty-seven fetal samples were obtained from women who received the prenatal cytogenetic examination for different reasons. In this study, the incidence of fetal chromosomal aberration related to each reason was analyzed. The incidence of de novo chromosomal aberration in the fetuses of the higher maternal age group was 1.5% (8/525: 4 cases of 21-trisomy, 2 cases of 18-trisomy and 2 cases of 47,XXY). The incidence in the group of women who had borne (a) chromosomally abnormal child(ren) was 0.5% (1/202). There was no repeat of 21-trisomy, 18-trisomy or 13-trisomy in this study. The incidence in the group of women who had ultrasonographic abnormalities was 16.9% (26/154). In this study, abnormal amniotic fluid volume, intrauterine growth retardation, malformation, hydrops fetalis, intestinal obstruction, omphalocele, single cord artery and cerebellar hypoplasia were related to chromosomal aberrations. The incidence in the group of women whose husband or herself was cytogenetically abnormal was 40.0% (26/65). This high value indicated that parental translocation and inversion are easily transmitted to their offspring.
Severe preeclampsia in the second trimester presents a major challenge in management because it is associated with high perinatal mortality. One possible way to improve the perinatal outcome in these pregnancies is to predict the later development of preeclampsia and to start management of it early. To investigate the relationship between the timing of the rise in maternal mean arterial pressure (MAP) and the course of biparietal diameter (BPD) growth in the fetus, we reviewed seven women with preeclampsia diagnosed between 18 and 27 weeks' gestation. MAP increased slowly within the normal range three or four weeks before the onset of the disease. Slowing of the fetal BPD growth curve preceded the onset of preeclampsia by two to four weeks. Serial measurement of BPD from early pregnancy may prevent the onset of severe preeclampsia before 28 weeks' gestation.