Biomedical subjects
Publications and source records attributed to T Kambara.
An alkaline hemoglobinolytic protease was extracted from the delayed hypersensitivity skin lesions induced by bovine gamma-globulin as an antigen in the guinea-pig. The enzyme was heat-labile and inhibited by thiol-blocking reagents. The mol.wt. was more than 100,000 and optimal pH around 9.
The effects of tetravalent conconavalin A and its succinylated derivative on the intracellular production of superoxide anion (O-2) and its release into cell exterior of peritoneal macrophages were observed. Both tetravalent concanavalin A and its succinylated derivative induced marked enhancement of intracellular reduction of nitroblue tetrazolium, which could be inhibited by alpha-methyl-D-glucoside. The extent of activation of nitroblue tetrazolium reduction induced by both types of the lectin paralleled the activation ratio of oxygen consumption. There was little difference in the extent of intracellular O-2 production induced by two types of the lectin. Nitroblue tetrazolium reduction was not affected significantly by pretreatment with colchicine, rotenone or malonate, inhibitors of the cytoskeletal system and of the electron transport system. In contrast, tetravalent concanavalin A induced a higher rate of superoxide release than did succinylated divalent concanavalin A, which lacks the cross-linking activity of surface glycoproteins. These results indicate that superoxide production following oxygen consumption and superoxide release into cell exterior are controlled independently by a separate membrane mechanism and that superoxide production system is not essentially dependent on the involvement of the cytoskeletal system.
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Conditions for the in vitro activation of the latent form of a protease-like permeability factor in the pseudoglobulin fraction from guinea pig skin were examined. (1) The factor was activated by dialysis against 67 mM phosphate buffer at pH 5.8--6.4, not at pH 7.0--8.0. (2) High salt concentration (200 mM or greater phosphate buffer or 67 mM phosphate buffer containing 200 mM or greater KCl or NaCl) prevented the activation at pH 6.2. (3) High osmotic pressure (sucrose at 1 M) did not affect activation at pH 6.2. (4) Reconversion of the activated permeability factor into an inactive form was not observed under high salt conditions, under which the latent permeability factor was stable in its own form. (5) The molecular size of the latent permeability factor was estimated as approx. 80 000 by Sephadex G-100 gel filtration at high salt concentration.
A permeability factor was extracted in a latent form from guinea pig skin and separated by ammonium sulfate fractionation into the pseudoglobulin fraction (30--50% saturation). The activation of the latent form of the permeability factor seemed to be caused in the desalting step by gel filtration with Sephadex G-50. The factor was partially purified by streptomycin treatment and column chromatography using hydroxyapatite, diethylaminoethyl cellulose and Sephadex G-75, in this order. Gel filtration showed that its molecular weight was approx. 35000. Its permeability activity was heat stable at 61 degrees C for 60 min at neutral pH, resistant at pH 5--10 and at ionic strengths from deionized water to 1 M NaCl at 4 degrees C. Its activity was transient and suppressed by guinea pig serum, but insensitive to an anti-histamic agent (triprolidine). Furthermore, its permeability activity was inhibited by diisopropylfluorophosphate, soybean trypsin inhibitor and leupeptin, and completely adsorbed by soybean trypsin inhibitor affinity column. These findings suggested that the permeability factor was a serine-type protease.
Partial purification of neutral proteolytic enzymes in guinea-pig lymphocytes yielded 2 enzymes. Both enzymes were heat-labile and inhibited by thiol reagents. The molecular weights were more than 200,000 and 150,000-200,000, and optimal pH around 9 and 8, respectively.
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Macrophage-chemotactic factors were extracted from delayed hypersensitivity skin lesions induced by bovine gamma-globulin in guinea pigs. The most active factor, MCFS--1, was highly purified and found to be a heat-labile protein with a molecular weight of 150,000 and to possess in vivo as well as in vitro activity. This factor was homogeneous during polyacrylamide gel electrophoresis, and the chemotactic activity was associated exclusively with this band. Further characterization revealed that its isoelectric point was 6.7 to 6.9 and made a single arc in the beta-globulin region with rabbit antiserums against guinea pig serum on immunoelectrophoresis. This factor seemed to be antigenically different from immunoglobulin G (IgG) by immunodiffusion and immunoadsorption. On the other hand, the chemotactic activity of MCFS-2 was adsorbed by neither anti-IgG nor anti MCFS-1 and that of euglobulin fraction was partially adsorbed by anti-IgG. These indicate the presence of at least three types of antigenically different chemotactic factors for macrophages in the extracts of delayed hypersensitivity skin lesions.
Eleven Macaca irus monkeys with a mixed or a permanent dentition were used in a study of the possible effects of extraoral forward force on the growth of the dentofacial skeleton. An intermittent forward force of 300 Gm. per side was applied, and the effects were analyzed by means of model casts, cephalometric radiographs, tetracycline bone marking, microradiographs, and histologic preparations. The results obtained were as follows: 1. An extraoral forward force caused significant changes in the circum-maxillary sutures and the maxillary tuberosity. 2. It was found that intermittent force of 300 Gm. per side resulted in desirable reactions in the sutures in young monkeys. 3. It was suggested that reactions in the suture might occur as a result of (1) an opening of the suture, (2) stretching of sutural connective tissue fibers, (3) new bone deposition along the stretched fibers, and (4) homeostasis which maintained the sutural width. 4. The maxillary complex showed a positional change with a little counterclockwise rotation. 5. A local reaction was observed histologically in the adult, but the degree was less than that seen in the young monkey. 6. It was considered that a forward movement of the maxillary complex would readily be accomplished by increasing sutural activity as early as possible. 7. Clinically, it was assumed that the hooks attached to the intraoral appliance should be placed as far frontally as possible. The present study seemed to indicate that forward movement of the maxillary complex would be performed with an extraoral anterior-pull appliance. Further investigation should be designed to evaluate a possible skeletal relapse after an orthopedic approach.
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A macrophage-chemotactic factor (MCFS) was extracted in the pseudoglobulin fraction from delayed hypersensitivity skin lesions induced by bovine gamma-globulin in guinea pigs. Its chemotactic activity was estimated by a modification of Boyden's method using Nuclepore filter. After chromatography of the protein fraction using Sephadex G-50 and DEAE-cellulose, in that order, two chemotactic fractions were obtained. The chemotactic factor with stronger activity (MCFS-1) was further highly purified (488-fold) by chromatography on CM-Sephadex. This factor migrated in a single band on acrylamide disc gel electrophoresis and was found to be a protein that was free of nucleic acid. Gel filtration showed that its molecular weight was similar to that of IgG. Its chemotactic activity was heat labile. Intradermal injection of this factor into normal guinea pigs induced a pronounced mononuclear cell emigration from venules. These findings are pertinent to understanding macrophage reaction in the delayed hypersensitivity reactions. Am J Pathol 87:359-374, 1977).