H mode observed in the JFT-2M tokamak with edge heating by electron cyclotron waves.
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Biomedical subjects
Publications and source records attributed to H Ohtsuka.
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The NADPH-dependent cytosolic 3,5,3'-triiodo-L-thyronine(T3)-binding protein (CTBP) has been purified over 30,000-fold from rat kidney by using charcoal extraction, Mono Q-Sepharose, Blue Sepharose CL-6B, and Sephacryl S-200 column chromatography. Purified CTBP had a sedimentation coefficient of 4.7 S, Stokes radius of 32.5A, and calculated molecular weight of 58,000. The apparently homogeneous protein consisted of a single polypeptide chain with Mr of 58,000 as estimated by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. Scatchard analysis of T3 binding showed that NADPH increases maximal binding capacity without changes in the affinity constant (Ka = 2.43 X 10(9) M-1). Double reciprocal analysis of NADPH and binding capacity gave maximal binding capacity of 16,400 pmol/mg of CTBP, Mr = 58,000. The order of affinity of iodothyronine analogues to purified CTBP was as follows: L-T3 = D-T3 greater than triiodothyroacetic acid greater than L-thyroxine. [125I]T3 bound to purified CTBP spontaneously dissociated from CTBP at 20 degrees C (t 1/2 = 22 min) in the absence of NADPH, whereas the dissociation was not observed in the presence of NADPH. The optimal pH for T3 binding was 7.2-7.5 Na+, K+, Ca2+, and Mg2+ (0-200 mM) did not influence T3 binding to CTBP. The purified CTBP did not bind to DNA and was not adsorbed to concanavalin A-Sepharose.
Cytosolic NADPH-dependent 3,5,3'-triiodo-L-thyronine (T3)-binding protein (CTBP) purified from rat kidney was further characterized in its T3 binding and its interaction with nuclei. Pretreatment of the CTBP with NADP induced dithiothreitol (DTT)-dependent T3 binding. The DTT-dependent T3 binding was increased by NADP in a concentration-dependent manner, and the maximal binding was obtained by 0.1 microM NADP. Higher concentrations of NADP (more than 0.1 microM), however, reduced T3 binding. NAD also induced DTT-dependent T3 binding, but was very low compared to that induced by NADP. NADPH and NADH did not produce DTT-dependent T3 binding. This NADP-activated, DTT-dependent T3 binding was characterized as follows: Ka for T3 binding was 1.8 x 10(9) M-1, and the maximal binding capacity was 15,000 pmol/mg of protein in the CTBP activated by 0.1 microM NADP. The molecular weight of the CTBP was 58,000 (4.7 S). A complex of [125I]T3 and CTBP (NADP.DTT.CTBP.[125I]T3), which was made from the CTBP pretreated with NADP and DTT, did not bind to DNA. However, the complex bound to the nuclei prepared from rat kidney. Treatment of the nuclei with 0.38 M KCl and with DNase I did not lead to loss of the binding activity for the complex. Treatment of nuclei with 0.5 M NaCl led to the loss of the activity for binding the complex. A complex of [125I]T3 and NADPH-activated CTBP did not bind these nuclear preparations. These results suggested that the active form of CTBP is present in two different forms: one is NADPH-activated, which plays a role as a reservoir for cytoplasmic T3, and the other is NADP-activated, which plays a role as a T3 carrier protein that transfers T3 from cytoplasm to nucleus.
Hybridoma cell lines producing human monoclonal antibodies (MAbs) MH-4H7 and KN-2B11 [immunoglobulin M (lambda)] which bound to the outer core region of Pseudomonas aeruginosa lipopolysaccharide (LPS) were established by cell fusion of human peripheral lymphocytes with human-mouse heteromyeloma SHM D-33. Both binding specificity experiments with a series of LPS-defective mutants derived from P. aeruginosa PAC1R (P. S. N. Rowe and P. M. Meadow, Eur. J. Biochem.132:329-337, 1983) and competitive enzyme immunoassay experiments with monosaccharides demonstrated that alpha-L-rhamnose residues in the outer core of LPS might be in part an epitope. The MAbs specifically bound to clinical isolates belonging to Homma serotypes A, F, G, and K at a frequency of 70 to 86% and to serotypes H and M isolates at about 50%. They did not bind to any isolates of serotype B, E, and I tested. This evidence indicates that L-rhamnose and probably its neighboring residues in the other core of P. aeruginosa are heterogeneous in some association with the O serotype.
The role of cytosolic T3-binding protein (CTBP) in the regulation of nuclear T3 binding was studied in vitro. Nuclear [125I]T3 binding was observed in the presence of 1.0 mM dithiothreitol (DTT). When the nuclei prepared from rat kidney were incubated with inactive form of CTBP which was also prepared from rat kidney, [125I]T3 binding to nuclei was not affected. When the nuclei were incubated with inactive form of CTBP in the presence of NADP, [125I]T3 binding to nuclei was increased, whereas binding was diminished when nuclei were incubated with CTBP in the presence of NADPH. The inactive form of CTBP was activated by NADPH. NADP also activated CTBP in the presence of DTT. Both active forms of CTBP were again inactivated by extraction with charcoal, and these inactive forms were reactivated by NADPH or by NADP and DTT, but not by NADP alone. Although the nuclei treated with 0.3 M NaCl lost the binding activity for [125I]T3 in the absence of NADP, the nuclei retained the binding activity for [125I]T3 in the presence of NADP and the inactive form of CTBP. Treatment of the nuclei with 0.5 M NaCl lost the binding activity for [125I]T3 not only in the absence but also in the presence of NADP and CTBP. These results suggested that NADP and NADPH play roles as counterregulatory factors for nuclear T3 binding in the presence of CTBP. Further, it was speculated that binding sites for the T3-CTBP complex, which is generated in the presence of NADP and DTT, are present in nuclei, and that binding sites for the complex are different from nuclear T3 receptors.
Effect of cytosolic 3,5,3'-triiodo-L-thyronine (T3)-binding protein (CTBP) on [125I]T3 binding to nuclei was investigated in vitro. CTBP and nuclei were prepared from rat kidney. CTBP was inactivated by incubating with charcoal and activated by incubating with NADPH or with NADP and dithiothreitol. Two complexes of CTBP and T3 [CTBP-T3(NADPH) and CTBP-T3(NADP)] were separately prepared, and the functions of these complexes were estimated. [125I]T3 binding to nuclei was not influenced by the inactive form of CTBP. NADP or NADPH alone did not modify [125I]T3 binding to the nuclei. However, the binding was markedly inhibited by NADPH in the presence of the inactive form of CTBP, but it was not inhibited by NADP in the presence of the inactive form of CTBP. The ability of nuclei to bind [125I]T3 was markedly diminished by pretreatment of the nuclei with 10(-8) M unlabeled T3. The diminished activity was not modified by adding NADPH or NADP. However, [125I]T3 bound to these nuclei in the presence of NADP and the inactive form of CTBP. Binding of [125I]T3 to these nuclei was not observed in the presence of NADPH and the inactive form of CTBP. When the nuclei that had previously been saturated with 10(-6) M unlabeled T3 were incubated with [125I]T3-CTBP(NADP) complex, radioactivity bound to the nuclei. The binding of radioactivity, however, was not observed when these nuclei were incubated with [125I]T3. The [125I]T3-CTBP(NADPH) complex did not bind to these nuclei. When the nuclei that had previously been treated with T3-CTBP(NADP) complex were incubated with [125I]T3, radioactivity bound to the nuclei. The binding of radioactivity, however, was not observed when these nuclei were incubated with [125I] T3-CTBP(NADP) complex. The [125I]T3-CTBP(NADPH) complex did not bind to these nuclei. These results suggested that 1) CTBP activated by NADP plays a role as a carrier protein for T3 from cytoplasm to nucleus; 2) there are binding sites for T3-CTBP(NADP) complex in rat kidney nuclei; and 3) these binding sites are different from nuclear T3 receptors.
The effects of histone subfractions on rat liver thyroid hormone receptor-DNA interaction were examined using an in-vitro DNA-cellulose binding assay. H1 histones bound to DNA showed reversible and potent inhibition of receptor-DNA binding without affecting receptor hormone binding. Poly-lysine, bovine serum albumin, ovalbumin and cytochrome c did not alter receptor-DNA binding. H1 histone subfractions (calf thymus lysine-rich histone (CTL)-1, CTL-2 and CTL-3) showed potent inhibition of receptor-DNA binding indistinguishable from each other. The quantity of H1 histone subfractions bound to DNA was the same. Although each subfraction has different functional properties, inhibition of receptor-DNA binding was a common feature of all the H1 histone subfractions, which is important for the non-random distribution of the receptor in chromatin. Binding of the receptor to core histones was investigated; it was found to bind to core histones more potently than to other proteins (H1 histone, ovalbumin and cytochrome c). Among core histone subfractions, H4 histone bound to the receptor most potently and is the candidate to be one of the acceptor sites of the receptor in chromatin.
A clinical trial was performed to examine the effect of ACDEMIN, a combination of lysozyme chloride and vitamins (manufactured by Grelan Pharmaceutical Co., Ltd.,; supplied by Takeda Chemical Industries, Ltd.). The subjects were 65 patients with slight to moderate symptoms associated with locally developed diseases including gingivitis, periodontitis, pericoronitis of the wisdom tooth and gingival abscess. Improvement of the condition was evaluated according to symptom on the basis of local findings examined prior to and 7 days after administration. Adverse effects were also evaluated in terms of discomfort. General improvement was determined on the basis of improvement in symptoms and general safety on the basis of a comprehensive assessment of the adverse effects. The usefulness of the drug was determined on the basis of general improvement and general safety as assessed above. The results were as follows: 1) Of the 65 patients who entered the trial, 62 completed the course of administration. 2) The rates of improvement ("slightly improved" or better) according to symptom were 65.6% for gingival inflammation, 40.0% for bleeding, 50.0% for pus discharge, 41.8% for swelling, 61.9% for local pain, 26.7% for mouth odor, 21.7% for color tone and 62.3% for discomfort. 3) The rates of usefulness ("slightly useful" or better) according to disease were 66.7% for gingivitis, 92.0% for periodontitis, 81.8% for pericoronitis of the wisdom tooth and 100.0% for gingival abscess. 4) The usefulness of the drug was graded "very useful" in 4 patients, "fairly useful" in 18, "slightly useful" in 31 and "not useful" in none, with an overall rate of usefulness of 85.5% ("faily useful" or better). 5) No patients presented symptoms indicating an adverse effect.
Response of the cervical sympathetic activity (CSA) to acute hypoxemia was studied in dogs in the absence and in the presence of either halothane (H), enflurane (E), isoflurane (I) or sevoflurane (S). Multifiber potentials of CSA, EEG, heart rate (HR), arterial blood pressure (ABP) and arterial oxygen saturation (SaO2) were continuously monitored. CSA increased responding to acutely induced hypoxemia at a speed of -6 - 9% (SaO2).min-1. The response of CSA was estimated in two ways as a function of SaO2: The threshold was determined at the SaO2 level where CSA increased by 3% over the basal level and the gain was determined by the slope of CSA increase divided by the SaO2 span. They were 92% and -1.88 in the absence of volatile anesthetics, respectively. The threshold and the gain decreased markedly in the presence of various concentrations of volatile anesthetics. They were 89.2% and -1.16; 81% and -0.74; 84.2% and -1.16 and 86.2% and -0.94 with 1 MAC of H, E, I and S, respectively. The depressant effects of volatile anesthetics on the CSA responses to acutely induced hypoxemia can cause depressed control of the autonomic nervous system against hypoxemia and conceal the manifestation of clinical signs of hypoxemia.
Inhibitors of erythropoiesis found in the blood of uremic patients have been implicated as some of the causes of anemia in chronic renal failure. As a treatment to remove the inhibitors, continuous ambulatory peritoneal dialysis (CAPD) is thought to be more effective than hemodialysis. Some researchers have reported that CAPD has improved anemic state in hemodialysis patients. However, there have been some cases of severe anemia seen in CAPD patients. To access the effect of the CAPD, we have performed serial examination on hemoglobin (Hb) and hematocrit (Ht) in 5 uremic patients under CAPD treatment. Hb levels in 4 and Ht in 3 of 5 patients have been known to be increased. One patient was shown to have no increment on the levels of Hb and Ht. On the other hand, further investigation has been down to access the effect of the recombinant human erythropoietin (r-HuEPO: Chugai pharmaceutical) on anemic patients, in whom no effect was noticed by treatment of CAPD. One severe anemic patient, who had been undergoing the treatment of CAPD for two years, was given r-HuEPO intravenously. The patient showed the increment on the levels of Hb and Ht, and the decreases on the levels of serum iron and ferritin. However, the patient has developed hypertension. In conclusion, CAPD was known to be not always effective for the increment of Hb and Ht, while r-HuEPO was very effective for the improvement of severe anemia in CAPD patients which enabls them to lead better social life for them.
One-stage eye socket and eyelid reconstruction was performed, with relatively satisfactory results, on a patient with a broad forehead. This was done after the complete exenteration of the orbit, using an island frontal flap and a retroauricular island flap with the common superficial temporal vessels. A Foley urethral catheter and a continuous suction drainage tube were effective for good wound healing between the bony orbit and a skin bag, made with a frontal flap, which was hidden in the orbital cavity. On the eighth postoperative day, the skin incision producing the false palpebral fissure was made, and it was enlarged slightly thereafter. A nylon implant was inserted into the orbital apex and cartilage grafts from the auricle to the false upper and lower eyelids were performed secondarily. The fundamental consideration of the eye socket and facial coverage corresponding to the eyelids and the possibility of modification of the author's method are briefly discussed.
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The effects of thyroid hormone on the levels of protein inhibitors for Ca2+-dependent proteinase were studied in rat brain. Four different inhibitory proteins (I, II, III, and IV) were isolated from immature (7-day-old) rat brain. The molecular weights of these inhibitory proteins, which were estimated by gel-exclusion chromatography on Sephacryl S-200 column, were approximately 280,000 (I), 70,000 (II), 50,000 (III), and 35,000 (IV). All of these inhibitory proteins were decreased by thyroidectomy. Four-day T4 administration (100 micrograms/kg daily) to thyroidectomized-immature animals restored proteins I, II, and IV. However, protein III was not recovered with the same treatment of the rats. In mature rats (40-day-old), four different inhibitory proteins were identified in the brain. The molecular weights were identical to those obtained in immature rat brain. As observed in immature rats, all of these inhibitory proteins were decreased by thyroidectomy. In contrast to the results obtained in immature animals, however, inhibitory protein III, as well as I, II, and IV, was restored by T4 administration (100 micrograms/kg daily) to thyroidectomized mature rats. The results suggested that thyroid hormone increases protein inhibitors for Ca2+-dependent proteinase in brain, and that irreversible decrease in one of the inhibitors is induced by thyroid hormone deficiency in immature animals. The phenomenon may be related to the neonatal hypothyroidism-induced irreversible damage to the central nervous system in patients with cretinism.
An aqueous two-phase partitioning study of partially purified nuclear thyroid hormone receptor from rat liver was performed. Stability of 3,5,3'-tri-iodo-L-thyronine (T3)-receptor complex and T3-binding activity in the presence of dextran or polyethylene glycol were assessed in order to determine the amount of occupied or unoccupied receptors in each phase. Partition coefficients were calculated as the ratio of receptor concentration in the upper polyethylene glycol-rich phase H2O and that in the lower dextran-rich phase H2O. The partition coefficient was a sensitive function of the salt at pH above 6.1 and below 5.1. The salt had no effect on the partition coefficient at pH around 5.6. These results suggest that the isoelectric point of the thyroid hormone receptor is about 5.6, confirming previous determinations using isoelectric focusing. The partition coefficient of the receptor decreased upon T3 binding, regardless of the salt composition. In contrast, the partition coefficient of thyroxine-binding globulin increased upon T3 binding. Free T3 preferentially partitioned into the upper polyethylene glycol-rich phase and gave a partition coefficient higher than 1.0. These results strongly suggest that the decrease in the partition coefficient of the receptor upon hormone binding reflects conformational changes or changes in electrostatic properties of the receptor upon hormone binding. Such an alteration may be involved in biological activation of the receptor upon hormone binding.
Estrogen receptor (ER) content in myoma and myometrium was determined by radioreceptor assay (RRA), and ER staining was performed by immunocytochemical staining, i.e., the peroxidase-antiperoxidase (PAP) method, in uterine samples from eight patients with uterine leiomyoma. ER content in myoma tended to be higher than that in myometrium, but the difference was not significant. The distribution pattern of ER staining was thought to be an important criterion. When the data were compared on the basis of the number of cells in the site where the largest number of positive cells aggregated, the number for myoma was significantly higher than that for myometrium (p less than 0.05). These results suggest that myoma contains more ER positive cells and more efficiently absorbs estrogen than myometrium.
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A 65-year-old woman visited our hospital, complaining dizziness and abdominal pain. Positive fecal occur blood and an iron-deficiency anemia were found. An investigation of the GI tract was performed, and enteroclysis revealed three stenotic lesions in the ileum, the most distal one having an irregular edge. A superior mesenteric angiogram showed tumor stain and encasement in the ileal region. On laparotomy, a large tumor and multiple stenosis were found in the ileum. Histological examination of the resected specimen confirmed the diagnosis of a carcinoid tumor and healed tuberculosis of the ileum.