Recently various component phenomena of the inflammatory reaction such as hyperemia, increasing vascular permeability, neutrophil infiltration, hemorrhage, and fibrin formation have been become measurable quantitatively using radio-labeled tracers, and have become induciable individually using purified or synthetic chemical mediators specific for each reaction. Using the techniques it has become increasingly apparent that the component of inflammation, which were considered as distinct processes, have stronger influences among them than previously appreciated. (1) It was previously noticed that when prostaglandins (E, I) were simultaneously injected with permeability inducing agents such as histamine and bradykinin, the plasma leakage caused by the agents was strongly augmented. In the recent works there have been shown many evidences that this effect of prostaglandins is attributable to hyperemia caused by them, and that this phenomenon actually occurs in several acute inflammation models. The mechanisms of the augmentation are regared as sum of effects such as increase of velocity of plasma efflux at each leaking site, increase of number of post capillary venules leaked, and increase of duration of the leakage at each site. The latter two were revealed by vital microscopic and electron microscopic observations of the microvasculature in hamster cheek pouch. (2) It has been indicated that neutrophil may be involved directly in the vascular permeability changes. For example, long lasting permeability reaction induced by chemotactic factors such as C5a, C5a des Arg, N-formyl-methionyl-leucyl-phenylalanine, and leukotreine B₄ is significantly reduced in neutrophenic animals, and the time course of the permeability change appeared to parallel that of neutrophil efflux. Furthermore, in acute inflammation models, participation of this type permeability reaction was observed. It is claimed that this type permeability change is hyperemiadependent or at least strongly augmented by it.
That these recent observations must make strong influences on the research for chemical mediation of inflammatory reaction, is finally discussed.

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About 100 years ago, Eli Metchnikoff had already expressed the importance of phagocytic cells in host defense. Today everyone admits that neutrophils are playing the most important role. Neutrophils migrate near the invading microorganisms, ingest them, kill them and digest them. Number and quality of neutrophils must be adequate in order to fulfill its function. Abnormal neutrophil function can be thought of as intrinsic cell defects, defects due to mediators acting on the cell and abnormalities related to shifts in populations of normal cells. The recent reviews of abnormal processes of neutrophil maturation, margination, chemotaxis, phagocytosis and killing will be explained in this capter.

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Cytotoxic activity of peripheral polymorphonuclear-leukocytes (PMN) of normal subjects and patients with various diseases was tested against K 562 cells for natural cytotoxicity and P 815 cells for ADCC by means of ⁵¹Cr-releasing assay incubated for 14 hours. Natural cytotoxicity of PMN of 20 normal subjects was 5.0% at a mean value. Mean values of PMN natural cytotoxiciy of patients with asthma, rheumatoid arthritis (RA), chronic hepatitis (CH), malignancy, Behçet's disease and SLE weel 10.8%, 4.7%, 1.2%, 1.8% and 1.5% respectively. ADCC activity of normarl PMN was 16.0% at a mean value. Mean values of PMN ADCC activity of patients with asthma, RA, CH, malignancy, Behçet's disease and SLE were 2.6%, 4.9%, 35.7%, 15.0%, 24.5% and 34.4% respectively. Natural cytotoxicity of PMN was around 1/6 of that of lymphocytes and ADCC activity of PMN was around 1/2 of that of lymphocytes. However, in all 4 patients with SLE and 2 of patients with Behçet's disease, ADCC activities of their PMN were higher than those of their lymphocytes.

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We evaluated halopredone acetate in basic and clinical aspects. Halopredone acetate is a highly topical corticosteroid. When it is used for intraarticular injections, the effects last longer than any other steroids which have been used, and it has less general effects. We applied halopredone acetate for RA and OA patients. For RA patients, mean dose for a wrist is 12.5 mg and that for a knee is 25 mg. About 90% cases showed effectiveness and in about 45% cases the duration of effect is longer than 4 weeks. More than half cases of OA showed also improvements.

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Leukocytes participate in inflammatory reaction by stimulus-linked release of granules and by synthesis of prostaglandins. Blood platelets are also circulating secretory cells and we have recently demonstrated that phenothiazines known as inhibitors of calmodulin inhibit stimulus-linked platelet reaction by blocking release of arachidonic acid from membrane phospholipids. Considering similarity of these cells, the effect of phenothiazines on the stimulus-linked release reaction of neutrophils was investigated. Rabbit neutrophils were obtained according to the method of Becker and Showell. Release reaction was triggered by addition of synthetic peptide FMLP to washed neutrophils pretreated with cytochalasin B. β-glucuronidase and lysozyme were measured as markers of lysosomal enzyme. Also lactate dehydrogenase (LDH) was assayed as an indicator of cellular destruction. 10 μM of FMLP was able to induce rapid release of two lysosomal enzymes assayed without significant release of LDH. About 60% of total enzymes were released within 1 minute after the addition of stimulus. Chlorpromazine hydrochloride (CPZ) or promethazine hydrochloride (PMZ) was used as an inhibitor of calmodulin, each of which dose-dependently inhibited the release of lysosomal enzymes. The complete inhibition of the release was observed by either 100 μM CPZ or 200 μM PMZ. In conclusion, we have demonstrated that phenothiazines inhibit FMLP stimulated release reaction of neutrophils, indicating the involvement of calmodulin in the reaction. The exact mechanism of the inhibition, however, has yet to be elucidated.

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A 35 year-old woman developed trigeminal neuropathy in the early course of mixed connective tissue disease (MCTD) . She had Raynaud's phenomenon, swollen finger, myalgia, muscle weakness, arthritis, skin eruption, oral sicca symptom, and unilateral trigeminal neuropathy. Serological study revealed positive antinuclear antibody of speckled pattern and solitary anti RNP antibody. These clinical manifestations and serological studies were compatible with MCTD. Trigeminal neuropathy in association with various connective tissue diseases were discussed. Trigeminal neuropathy was thought to be a rare condition in conective tissue diseases such as sclero derma, polymyositis, systemic lupus erythematosus, and so on. But, this condition had been seen more frequently in the overlapping type of some connective tissue diseases, especially in MCTD.

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Possible presence of slow reacting substance (SRS) in the synovial fluid of rheumatoid arthritis was studied. 15 samples out of 30 (50%) of synovial fluids showed SRS-like smooth muscle contractile activity ranging from 1.4 to 143 units/ml. This SRS-like activity on the guinea pig ileum was reversed by the specific SRS-A antagonist FPL 55712, and was hydrolysed by both purified human placental arylsulfatase B and type IV limpet arylsulfatase. Elution profiles of SRS-like substance on silicic acid chromatography, Sephadex LH-20 and DEAE-Sephadex A (-) 25 column chromatography were compatible with those of SRS-A. These results suggested that SRS was released from some kind of cells, possibly polymorphonuclear leukocytes, which migrated and stimulated immunologically in the synovial fluid of rheumatoid arthritis patient, and it might play any pathophysiological roles in the disease.

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A method to assay the level of neuraminidase in human plasma was developed. The synthetic fluorescent substrate N-acetyl-neuraminic-α-methylumbelliferil ketoside developed by Dr. M. Flashner of Syracuse, New York was used. Twenty μl of plasma and 200 μl of 60 μ mol of substrate in sodium hosphate buffer (pH 5.8) were mixed and incubated at 37°C for two hours. The rcaction was stopped by using 2, 500 μl of 10% sodium carbonate. The level of neuraminidase activiy was determined by measuring the fluorescence of methylumbelliferone which had been liberated by the activity of neuraminidase on the substrate. A fluorescent spectrophotometer (JASCO FP 550) using a wave length of 327 nm for excitation and an emission of 448 nm was used. Using this assay, Km was found to be 20μ mol 1 and the minimum measurable level of neuraminidase activity was 10^-5U/ml. The assay was not influenced by protein in the plasma. This assay makes possible the measuring of a small amount of neuraminidase activity in the plasma.

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Rat pleurisy was induced by intrapleural injection of 2 % λ-carrageenin. Complement titers were measured by Lachmann's method (50% hemolysis, CH 50), using EAC142, which was made by preincubation of erythrocytes with zymosan-treated serum. In vitro incubation of 0.2 and 0.4% carrageenin with serum decreased the residual CH50 values doseand time-dependently. Complement titers in pleural exudate, expressed in terms of mg protein, was lower than those in serum in the whole course and it was concluded that complement was activated in the pleural cavity. Polymorphonuclear leukocytes (PMN) increased their numbers rapidly up to 5 hr and stayed at the same level thereafter, whereas mononuclear cell number increased gradually up to 24 hr. Using dexamethasone [sharing arachidonic acid (AA) release inhibition], benoxaprofen (a lipoxygenase inhibitor with a weak cyclooxygenase inhibitor) and K-76COONa (an inhibitor of complement activation), it could be concluded that PMN migration during the first 3-7 hr may be attributable to complement and AA metabolites, but no involvement of AA metabolites was conceivable in the 14-19 hr pereiod. Mononuclear cell migration in the beginning (3-5 hr) might not be induced by neither complement nor AA metabolites, except 7 hr, in which lipoxygenase products might be involved. Benoxaprofen inhibited mononuclear cell migration in 14-19 hr period.

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Anti-inflammatory, analgesic and ulcerogenic activities of CS-600 and its active metabolite were investigated in rats. Anti-carrageenin edema activity of CS-600 (ID₅₀: 1.2 mg/kg) was as potent as that of the active metabolite (ID₅₀: 1.3 mg/kg) . Analgesic effect of CS-600 (ID₅₀: 0.76 mg/kg) on scald pain was also equipotent to that of the metabolite (ID₅₀: 1.1 mg/kg) . However, gastrointestinal irritancy of CS-600 (UD₅₀: 16.1 mg/kg for the stomach and 11.5 mg/kg for the intestine) was lower than that of the active metabolite (UD₅₀: 3.2 mg/kg for the stomach and 3.1 mg/kg for the intestine) . When [¹⁴C] CS-600 was injected into a ligated jejunum in rats, most radioactivity was found as the unchanged CS-600 in the portal blood. These findings indicate that CS-600 is absorbed from the intestine in the less irritating form and that it is converted rapidly to the active metabolite which shows the potent pharmacological efficacy.

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Effects of CCA (disodium 4-chloro-2, 2'-iminobenzoate) on the production of plaque forming cells (PFC) in murine spleens were investigated and the following findings and results were obtained in this paper. (1) CCA had almost no effect on anti SRBC PFC responses at optimal experimental conditions in normal animals but had at some suboptimal condi-tions. (2) In T cell-deficient nude mice, CCA increased the percent of Thy-1⁺ cells in the spleen and enhanced the product ion of PFC to T-dependent antigen such as SRBC. CCA also showed an enhancing effect on the production of PFC to T-independent antigen, LPS. (3) Induction of suppressor T cells by Con A in the spleen of NZB/W F₁ mice in which the activity of these cells are known to be decreased was enhanced by the treatment of CCA. (4) From the above findings. CCA seems to affect both helper and suppressor T cells and B cells. 5 Such a multiple action of CCA on lymphocytes populations may explain the mechanism by which CCA does not affect the normal immune responses but does the hyper-or hypo-immune responses resulting from the deficiency and the impainment of a specific lymphocyte population as observed in the case of nude mice or NZB/W F₁ mice.

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Anti-inflam-matory mechanism of butyl flufenamate (HF-264) was studied by various analysis. HF-264 did not inhibited the heat denaturation of bovine serum albumin, whereas flufenamic acid showed a marked inhibition on it. HF-264 and flufenamic acid prevented the dog erythrocytes from heat induced lysis, and inhibited the production of superoxide anion in macrophages. HF-264 inhibited the synthesis of prostaglandin E₁, but its inhibition of HF-264 on it was more weaker than that of flufenamic acid. From the above results, it is suggested that some part of potent anti-inflammatory action of HF-264 is the results of the interaction of HF-264 with biomembrane, as membrane stabilization and inhibition of superoxide anion production.

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