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Biomedical subjects

T Horio

Publications and source records attributed to T Horio.

At least 217 records · Page 12Linked to original sources

Alteration of lymphocyte functions by 8-methoxypsoralen and long-wave ultraviolet radiation. II. The effect of in vivo PUVA on IL-2 production.

Our previous studies demonstrated that psoralen plus long-wave ultraviolet radiation (PUVA) treatment inhibited certain T-lymphocyte functions, such as locomotive ability. To further analyze the effects of PUVA on T-lymphocyte function, we investigated the ability of mouse spleen cells to produce interleukin 2 (IL-2) after treatment of the mice in vivo with PUVA. Interleukin 2 production was impaired in cells from PUVA-treated mice compared with those from UVA-irradiated, 8-methoxypsoralen-treated, or normal mice. This impairment was not dose dependent, over the dose range of UVA (2-20 J/cm2) examined. Interleukin 2 production was markedly suppressed on day 3 after PUVA and returned to normal by day 7 after the treatment. Topical treatment of the mice with croton oil did not affect IL-2 production of their spleen cells. This result indicates that cutaneous inflammation per se may not be responsible for the suppressive effect of PUVA on IL-2 production. Addition of exogenous IL-1 did not reconstitute the decreased ability of spleen cells to produce IL-2 in vitro, indicating that PUVA affected primarily IL-2 producing cells. These suggest that impaired IL-2 production may account for some of the immune dysfunction observed in PUVA-treated animals.

Animals↗

Purification of acetylcholine receptor-like protein from fetal calf thymus.

A cobrotoxin binding protein from the fetal calf thymus was isolated by affinity chromatography after solubilization with sodium cholate. The specific activity as a nicotinic acetylcholine receptor (AChR) was determined by assessing the binding to [3H]-alpha-bungarotoxin (BuTx), using the high-pressure liquid chromatography. An AChR-like protein was detected in the amount of 1.39-2.14 nmol per g protein. The first peak of 420k-protein from gel filtration of the eluate of affinity chromatography on a Sephacryl column showed one major polypeptide band with an Mr of 40k, by polyacrylamide gel electrophoresis in sodium dodecylsulfate, two major protein bands with pI 5.4-5.6 and 9.2 by isoelectric focusing, and reacted with sera from patients with myasthenia gravis.

Animals↗

Occupational dermatitis with simultaneous immediate and delayed allergy to chrysanthemum.

A 42-year-old woman had recurrent dermatitis on the hands, arms, face and trunk for 5 years. She was allergic to chrysanthemum, with positive patch tests. She also had contact urticaria to chrysanthemum, with a wheal-and-flare reaction 15 min after application of chrysanthemum juice to a scratch test site. This was not seen in control subjects. The co-existence of allergic contact dermatitis and contact urticaria from chrysanthemum might play a rôle in the development of dermatitis, especially severe chronic dermatitis of her hands.

Adult↗

Histogenesis of UV light-induced blister formation.

Histological examination of ultraviolet light (UVL)-induced blister formation in guinea pig skin is reported. Light and electron-microscopic examination was performed on skin exposed to single and multiple exposures of a large dose of UVB. In light microscopy, intracellular edema was prominent, especially in the lower part of the epidermis, where it developed into a blister. It seemed to be a subepidermal bulla. The PAS-positive basement membrane was observed at the floor of the blister. The positive Nikolsky's sign seemed to be due to a separation between the degenerated and regenerated epidermis. Electron microscopy revealed a marked intercellular as well as intracellular edema and well-preserved desmosomes. The basal lamina was always located at the floor of the blister. The difference in UVL-induced blister formation between normal and pathological conditions is discussed.

Animals↗

Preliminary crystallographic study of a ribulose-1,5-bisphosphate carboxylase-oxygenase from Chromatium vinosum.

Crystals of a ribulose-1,5-bisphosphate carboxylase-oxygenase from Chromatium vinosum were obtained with the hanging-drop vapor diffusion technique, using polyethylene glycol 4000 as precipitant. The crystal belongs to the cubic system, space group I432, with unit cell dimension a = 245.9 A. An asymmetric unit includes one-quarter (L2S2, L: large subunit, S: small subunit) of a hexadecameric molecule (L8S8, 544,000 Mr), which is located on the crystallographic point symmetry 222 or 4. The crystal diffracts to at least 3.0 A resolution.

Chromatium↗

A functional study of a case of glucagonoma exhibiting typical glucagonoma syndrome.

A 46-year-old man had a 7-year history of severe rash, which was then diagnosed as necrolytic migratory erythema. He had a weight loss of 6 kg, abnormal glucose tolerance test findings, anemia, glossitis, hair loss, and hypoproteinemia. Plasma amino acids levels were significantly decreased, and the fasting plasma glucagon (IRG) level was high at 5000 to 8000 pg/ml. Circulating IRG significantly increased after oral glucose loading, meal ingestion, and arginine infusion, and decreased with somatostatin infusion and insulin-induced hypoglycemia. No other gut or pancreatic hormone levels in plasma were elevated. Plasma IRG was eluted by gel-filtration, mainly in the position of true glucagon (MW 3500) by antiserum 30K. The rash was markedly improved after infusion of amino acids. Computerized tomography (CT) scan and celiac angiography revealed a large pancreatic tumor with multiple liver and lymph node metastases. The pancreatic tumor was totally resected, and was identified as glucagonoma by immunohistochemical technique. Since the plasma IRG levels remained high after surgery, the patient received dimethyltriazenoimidazole carboxamide therapy. After several courses of this treatment, plasma IRG levels decreased to 1000 to 2000 pg/ml, and the hepatic metastases were remarkably diminished in size.

Adenoma, Islet Cell↗

The inhibitory effect of PUVA on the immunity of experimental dermatophytosis in guinea pigs.

The effect of topical PUVA on the disease course and immunity of T. mentagrophytes dermatophytosis was investigated in guinea pigs. Animals which had been inoculated on nontreated skin showed mild erythematous lesions with scaling in a few days and then developed the most intense reaction between days 10 and 14. The lesions resolved completely by the third week. On the other hand, animals which had been inoculated on the PUVA-treated sites showed only mild squamous, erythematous lesions until the fourth postinfective week, when the intense reaction began to appear. Complete regression was observed by the fifth week in these animals. Trichophytin tests performed on the 14th day were positive in the guinea pigs of non-treated group, while negative in the PUVA-treated animals. The latter group revealed a positive reaction on the fifth week. PUVA did not show inhibitory effect on the sensitization by intracutaneous injection of trichophytin antigen. The PUVA treatment depleted the ATPase-positive Langerhans' cells. These results indicate that PUVA treatment suppresses the immunity of dermatophytosis and delays the spontaneous resolution of the lesions, and suggest that the Langerhans' cell is involved in the development of cell-mediated immunity in experimental dermatophytosis.

Animals↗

Ear swelling in response to UVB irradiation.

The skin response to UVB irradiation in mice was evaluated by means of ear swelling. ICR albino mice were irradiated with 500 mJ/cm2 UVB and the effect of various drugs on ear swelling was examined 24 h after irradiation. Intravenous injections of betamethasone (0.8 microgram/g body wt.) or indomethacin (24 micrograms/g) remarkably inhibited ear swelling, whereas intraperitoneal injections of diphenhydramine (20 micrograms/g), cimetidine (10 micrograms/g), or a combination of both these antihistamines did not. In contrast, the number of sunburn cells counted 24 h after UVB irradiation (200 mJ/cm2) in mouse ears was not affected by these drugs. The amount of prostaglandin D2 (PGD2) in mice ears at various intervals after irradiation with 500 mJ/cm2 UVB was determined by radioimmunoassay. Compared with the values before irradiation, the PGD2 levels were significantly higher 3 and 6 h after irradiation and gradually decreased and returned to the basal level by 12 h, although ear swelling continued after 12 h. These results suggest that prostaglandins are responsible at least in part for the development of ear swelling, but not for sunburn cell formation induced by UVB, and that ear swelling represents a simple and useful response model for the rapid in vivo screening of nonsteroidal or steroidal anti-inflammatory agents.

Animals↗

Multiple unit activities recorded from the medial preoptic area during copulatory behavior in freely moving male rats.

Multiple unit activities (MUAs) were recorded from the medial preoptic area (MPO) during copulatory behavior in freely moving male rats. The baseline firing rate in the male MPO was 19.6 spikes/s +/- 7.1 S.E. (n = 14), and it was increased by 36.7% when a female rat was introduced (26.8 spikes/s +/- 8.9 S.E., n = 14). The firing rate remained elevated until ejaculation. The maximum firing rate (53.5 spikes/s +/- 16.7 S.E., n = 14) was obtained specifically during pursuit-mounting behavior. This high frequency firing with pursuit-mounting behavior was, however, immediately inhibited for 8.2 s +/- 2.1 S.E. (n = 14) when mounting was followed by intromission. The firing-inhibition was further prolonged (120.9 s +/- 44.6 S.E., n = 14) when ejaculation was performed after mounting and intromission. In cases where mounting was elicited alone, a firing-inhibition was not observed and the firing rate after mounting returned to the average firing rate during copulatory behavior. These results suggest that the MPO neurons may play a major role in pursuit-mounting, intromission and ejaculation.

Action Potentials↗

Purification and properties of epithelial growth inhibitor (EGI) from human platelets: its separation from type beta transforming growth factor (TGF-beta).

We previously reported that sera from various kinds of animals contain a protein(s) capable of inhibiting the growth of the non-malignant epithelial cell line derived from Buffalo rat liver (BRL). In the present study, a similar epithelial cell-specific growth inhibitor (EGI) was purified to homogeneity from an acid-ethanol extract of human platelets. During purification, EGI was separated from the major component of type beta transforming growth factor (TGF-beta), which can stimulate the colony formation of the non-malignant fibroblastic cell line derived from rat kidney (NRK) in soft agar in the presence of epidermal growth factor (EGF). The purified EGI had an Mr of 27,000, and was composed of two subunits identical in Mr. It significantly inhibited the growth in monolayer cultures of three non-malignant epithelial cell lines, BRL, MDCK (from Madin-Darby canine kidney) and BSC-1 (from African green monkey kidney), at doses lower than 40 pg/ml in medium containing 10% fetal calf serum. Its inhibitory activity was stable against heating at 90 degrees C for 3 min, but not against treatment with 50 mM dithiothreitol. In addition, TGF-beta was also partially purified from the same extract. The purified TGF-beta did not show any inhibitory activity toward the growth of BRL, MDCK, BSC-1, or NRK.

Blood Platelets↗

Intermolecular relations of the photosystem II complex in spinach chloroplasts as detected by immunochemical assay.

Polyclonal antibodies were prepared to the subunits of the spinach photosystem II fraction (PS II): p47, p43, p27, p33, p24, and p17. (The protein nomenclature refers to Mr). p47 and p43 are the subunits of reaction center complex, and p27 is light-harvesting chlorophyll protein. p33, p24, and p17 are extractable from PS II with 1 M Tris, and p24 and p17 with 1 M NaCl. With untreated PS II fractions, the antibody to p24 inhibited the photosynthetic oxygen-evolving activity, but not the DCPI-photoreduction activity in the presence of DPC, indicating that p24 played an important role in the former activity. Bindings of the respective antibodies to the PS II treated with sodium dodecyl sulfate were regarded as 100%. To untreated PS II, the bindings were 20-30% for p47, p43, and p27, about 50% for p33, and 70-80% for p24 and p17. To NaCl-washed PS II, the binding to p33 increased by 9%, indicating that p33 was adjacent or bound with p24 or/and p17. To Tris-washed PS II, the binding to p43 increased by 7%, indicating that p43 was adjacent or bound with p33. To PS II treated with 3% of Brij 58, only the binding to p27 increased appreciably. To PS II treated with 1% of octyl glucoside, the binding to p47 was still lower than 50%, whereas those to the other subunits were 74-91%. These values could be a measure of the extents to which the subunits were exposed to the aqueous phase, because of the nature of polyclonal antibodies. These results suggest that in intact PS II, p47, p43, and p27 were in most part buried in the inside, p47 being located at the most central and p27 at the outermost part, whereas p33, p24, and p17 were exposed to the outside by 50-75%.

Antibodies↗

Effects of NaCl and glycerol on photosynthetic oxygen-evolving activity with thylakoid membranes from halophilic green alga Dunaliella tertiolecta.

It is known that the halophilic green alga Dunaliella tertiolecta grows under hypertonic conditions (with NaCl), which induce the intracellular accumulation of high concentrations of glycerol in order to counterbalance the osmotic change. The effects of NaCl and glycerol on the photosynthetic oxygen-evolving activity of thylakoid membranes prepared from D. tertiolecta were investigated in relation to the dissociation of the membranes. It was found that proteins with Mr of 24,000, 17,000, and 13,000 were dissociated from thylakoid membranes of D. tertiolecta by washing with 1 M NaCl, whereas the photosynthetic oxygen-evolving activity was stimulated 2-fold by 0.1-1.5 M NaCl. The antibodies against spinach 24K and 17K proteins did not cross-react with Dunaliella 24K and 17K proteins, respectively. The salt-tolerant feature of the oxygen-evolving activity with Dunaliella thylakoid membranes may be related to the difference of the properties of these two proteins between D. tertiolecta and spinach. When the membranes were washed with 1 M Tris, proteins with Mr of 50,000 and 31,000 were also dissociated in addition to the 24K and 17K proteins described above. The antibody against spinach 33K protein cross-reacted with 31K protein of D. tertiolecta, showing that Dunaliella 31K protein corresponds to spinach 33K protein. When the membranes were treated with a mixture of 1% cholate and 2% deoxycholate, the oxygen-evolving activity was completely depressed, but the depressed activity was significantly restored by organic solvents. Glycerol and dimethylsulfoxide were the most effective for the restoration.(ABSTRACT TRUNCATED AT 250 WORDS)

Chlorophyta↗

A novel FAD-protein that allows effective reduction of methyl viologen by NADH (NADH-methyl viologen reductase) from photosynthetic bacterium, Rhodospirillum rubrum: purification and characterization.

It was found that the cytoplasm of light-grown cells of Rhodospirillum rubrum could catalyze the reduction of methyl viologen (MV) (Em, 7 = -0.44 V) by NADH and NADPH. In the present study, the enzyme capable of catalyzing MV reduction by NADH (NADH-MV reductase) was purified 1,500-fold from an extract of cells with a yield of 4.4%. The purification procedure comprised (NH4)2SO4 fractionation, and chromatographies on Sepharose CL-6B, DEAE-Sepharose CL-6B, phenyl-Sepharose CL-4B, Blue-Cellulofine, and TSK-Gel G3000SW. Two NADPH-MV reductases were separated during the purification. The NADH-MV reductase obtained was nearly homogeneous, as judged on polyacrylamide gel electrophoresis both in the presence and absence of sodium dodecyl sulfate. The enzyme has a molecular weight of 220,000 and an isoelectric point of 4.8; it is composed of four subunits with a molecular weight of 57,000, and is bound with about 1 mol FAD/mol subunit. The activity is optimum at pH 8. The Km values for NADH and MV are 115 microM and 1.3 mM, respectively, with a molecular activity of 13,000 min-1. The activity was stimulated 2.4-fold in the presence of 20-100 mM ammonium ions. The enzyme also catalyzed the reduction of benzyl viologen, methylene blue and 2,6-dichlorophenol-indophenol (Em, 7 = -0.36, +0.011, and +0.217 V, respectively) at comparable rates. The ratios of the activity with NADH to that with NADPH were 80, 133, 41, and 5.5 with MV, benzyl viologen, methylene blue and 2,6-dichlorophenolindophenol, respectively. The enzyme was significantly stable in the presence of both 5mM 2-mercaptoethanol and 20% (w/v) glycerol. The activity was not appreciably influenced by the presence of 2 M urea, although the reagent caused dissociation to the subunits.

Chromatography↗

Barley leaf peroxidase: purification and characterization.

Peroxidase was prepared from extracts of barley leaves and separated into seven components, different in pI. The purification procedure comprised two parts. The first part was based on the fact that all the components had practically the same molecular weights. It consisted of fractionations with acetone and ammonium sulfate, ion-exchange chromatographies on CM-cellulose and DEAE-Sepharose CL-6B, and molecular-sieve chromatography on Ultrogel AcA44; the components were all purified together to near homogeneity on sodium dodecyl sulfate (SDS)-polyacrylamide gel electrophoresis, and the procedure resulted in 1,200-fold purification with a yield of 39%. The ion-exchange chromatographies were carried out under conditions such that the components would not be adsorbed. In the second part, the enzyme preparation was separated into the seven components by repeating isoelectric electrophoresis. Their isoelectric points (pI) were 6.3, 6.8, 7.4, 8.3, 8.5, 8.7, and 9.3. The components other than the pI 6.3 and 6.8 components were each purified to homogeneity in the electrophoresis. The seven components thus prepared were the same in molecular weight on SDS-gel electrophoresis (44,000) and showed absorption maxima at the same wave-lengths (403, 496, and 534 nm), RZ (A403/A275) ranging from 2.09 to 2.81. Their protoheme IX contents were 0.81-1.07 mol/mol, and their true sugar contents 15-26% (g/g). The amino acid compositions suggest that the five components described above are not real isoenzymes, but exhibit different pI values due to differences in glycosyl residue. The pI 9.3 component was crystallized in spite of its high sugar content.

Crystallization↗