After intravenous injection of colloidal carbon, subcutaneous injection of bradykinin (BK, 5-5000 ng/50μl) into the scrotum of rats induced carbon deposit along the venular trees of the cremaster muscle, since BK induced the gap formation between endothelial cells of venules. Endothelial cells of the collecting venules were most sensitive to BK and kept the gaps open longer than 20 min, whereas venules upstream with a diameter of about 10μm were less sensitive and closed the gaps within 5 min. Thus, individual endothelial cells show different sensitivity along the venular trees within a distance of 400 to 500 μm. Vital microscopy of microcirculation of hamster cheek pouch disclosed that polymorphonuclear leukocytes (PMNs) cross the endothelial layer to the interstitial space by four steps; adhesion to endothelial cells, penetration between adjacent endothelial cells, stay beneath the pericyte basement membrane (BM) and cross the pericyte BM. The experiments revealed that the adhesion of PMNs by LTB₄ was attributable to change in the PMN membrane and not to that in the vascular endothelial cells. Thus, the endothelial cells seem to take a passive role to the PMN penetration. Rapid and magnificent development of the techniques in culturing endothelial cells must be supplemented with the knowledge, which was aquired by in vivo observation and precise analyses of the nature of the endothelial cells in living animals under the microscope.

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When the cytokines, such as interleukin-1 (IL-1) and/or tumor necrotizing factor (TNF), are inocculated into the culture of human endothelium, the tissue plasminogen activator is lower and the tissue plasminogen activator inhibitor (PAT-1) is higher in the supernatant of the culture system than without inocculation. In addition, tissue type of coagulation factor is increased, suspecting the acceleration of fibrin formation in the course of inflammation. On the other hand, the plasminogen activator, which is an UK type activator, is derived from the monocytes/macrophages by the stimulation of substances such as thioglycolate, endotoxin and others which are related to the inflammation. Fibrinolysis and coagulation in inflammation are not necessarily unipolar. We need further information to be able to adequately explain the changes of tPA in Kawasaki disease which is one of the most severe vasculitis disorders in infants.

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Effect of AA861, a 5-lipoxygenase inhibitor of arachidonic acid, on natural killer activity in rat whole spleen cells (WSC-NK activity) or rat non-adherent spleen cells (NASC-NK activity) was examined. AA861 significantly depressed WSC-NK activity. On the contrary AA861 showed no inhibitory effect on NASC-NK activity. The depressed WSC-NK activity by AA861 was partly recovered by exogenously added leukotriene B₄ (LTB₄) in a dose dependent manner, with a significant recovering effect at 10^-10 M of LTB₄. Then we compared the capacity of LTB₄ production between WSC and NASC when stimulated with calcium ionophore A23187. WSC produced a significant amount of LTB₄, howevr, NASC produced only a negligible amount of LTB₄. We detected LTB₄ in the assay medium of WSC-NK activity and the amount of LTB₄ in the medium was decreasd by AA861 in a dose dependent manner. The current results may indicate that the inhibitory effect of AA861 on WSC-NK activity is partly mediated by the inhibitory effect of AA861 on LTB₄ production in rat spleen adherent cells.

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Renal scarring is thought to be end-stage of chronic pyelonephritis and one of important causes of renal insufficiency and renal hypertension. Significance of bacterial pili was examined in scar formation by the use of artificial variants which possessed either MR or MS pili of S. marcescens, and the result described below was obtained. MS piliated bacteria stimulated renal scarring more severely than MR piliated bacteria. This scarring mediated by MS piliated bacteria was prevented by superoxide dismutase.
These findings suggested that the MS pili stimulated polymorphonuclear leukocytes to release large amount of superoxide and this superoxide damaged the renal tissue, resulted in the renal scar.

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Contributions of prostaglandin (PG) s, thromboxane (TX) s, and leukotriene (LT) s in the pathophysiology of bronchial asthma have not been clarified yet. In the experimental model of bronchial asthma, SRS-A and histamine are known to play an important role. But clinical analysis is rare. In this investigation we performed broncho-alveolar lavage (BAL) in bronchial asthma patients and measured chemical mediators in three fractions of BAL fluid (BALF) and peripheral venous blood.
TXB₂ and 11-dehydro-TXB₂ were significantly increased in the BALF of patients with bronchial asthma. But their concentrations in the peripheral venous blood are not so high compared with those of control groups. PGD₂ and PGF_2α were not detected in the peripheral venous blood, but their levels in BALF were elevated in BALF of patient with bronchial asthma. Levels of leukotriene C₄ were not elevated in BALF and peripheral venous blood compared with those of control group, but further study would be needed to evaluate the role of leukotrienes including LTC₄ and LTD₄.

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To clarify a role of lysosomal cysteine proteinases, cathepsins B, H and L in periodontal tissue destruction, levels of their activities in gingival crevicular fluid (GCF) from patients with chronic adult periodontitis were determined using 4-methyl-7-coumarylamide substrates. All of the enzyme activities were positively correlated with the GCF volume, but not with the probing depth. Higher levels of the respective activities were found at sites with more serious signs of periodontal disease. The specific activity of each enzyme in GCF, however, was progressively decreased as the GCF volume increased, indicating the negative correlation between GCF volume and each enzyme activity. The results also suggest that cathepsins B, H and L are selectively released into gingival crevices at a mild stage of the periodontitis and that the levels of these enzymes in the GCF may be clinical parameters that reflect the degree of the periodontal tissue destruction.

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The concentration of medullasin, a granulocyte elastase-like serine proteinase, in gingival crevicular fluid (GCF) was determined by sensitive immunoassay method. In patients with chronic adult periodontitis the medullasin content was positively associated with the quantity of GCF, but not with the probing depth. The medullasin content was greatly decreased under the periodontal treatment, as compared with prior to the treatment. In individuals with experimental gingivitis the medullasin content in gingival crevices showed three distinct exudative patterns. The medullasin content in GCF attained a peak value at around 4 days after refraining from oral hygiene measures. It showed a rapid decrease up to 7 days followed by a progressive increase during a 21-day period. The increased medullasin was rapidly decreased by cleaning the teeth. The results suggest that medullasin play roles in development of both gingivitis and chronic periodontitis.

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The quantitative measurement of rheumatoid factor (RF) was made by laser nepherometry in order to examine the changes of RF in rheumatoid patients before and during antirheumatoid medication with auranofin, gold sodium thiomalate (GST) and CCA.
The patients were divided into 4 groups: 20 cases of auranofin group, 6 of GST group, 9 of CCA group, and 20 of control group. RF was measured with latex photometric immunoassay (Eiken Kagaku, LA-200) . Duration of observation were 41±25 weeks (mean±SD) on auranofin group, and 63±72 on control group. The RF rates of auranofin group and GST group were significantly improved with the results of 24.1±36. 1 (mean±SD) in auranofin group, 52.1±40.3 in GST group, in comparison with control group with the results of -2.4±36.1 (P<0.05, P<0.01) .
With these results, we concluded that quantitative measurement of RF is very useful for evaluating the efficiency of antirheumatic drugs, as it indicates the condition of rheumatoid arthritis.

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The mitogenic activity of TJ-48 in murine lymphoid cells was examined to get a clue for the augmentation of antibody production by TJ-48.
Mitogenic activity of TJ-48 was detected in spleen cells, lymphonodus cells but not in thymocytes. The effect was abolished by pretreatment with anti-Ig antibody but not affected by pretreatment with anti-Thy 1.2 antibody. Coculture of spleen cells with TJ-48 resulted in increased number of sIgM-, sIgG- or sIgD-positive cells according to FAGS analysis. The increase in mitosis by TJ-48 was abolished by the elimination of adherent cells and the readdition of adherent cells recovered the effect of TJ-48. These data indicate that TJ-48 is a B cell mitogen and the activity is T cell-independent and adherent cell-dependent. The mitogenic activity of TJ-48 may account for the augmentation of the immune response.

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Lobenzarit-2Na (CCA) is a new antirheumatoid arthritis agent and is reported to show an immuno-modulatory effects clinically and pre-clinically. Immunological modification by CCA in vivo and in vitro is so complicated, and it is difficult to expect CCA to exert its effect by activating or inhibiting a specific enzyme (s) . Actually it was reported that CCA had no inhibitory effect on cyclooxygenase.
Previously it was shown that RNA polymerase activity in nuclear suspension isolated from lymphonode was activated in a double-peaked fashion by CCA. A detailed study on RNA polymerase-I activity in nuclear suspension isolated from rat liver indicated an indirect effect of CCA on RNA polymerase-I, as described in this report. We deduce here that CCA affects the reactivity of cytoskeleton, which attaches periphery of nucleus, and that the organization state of cytoskeleton in nuclear suspension is responsible for the regulation of nucleolar enzyme, RNA polymerase-I activity.
This and following reports give a strong suggestion that CCA can modify the reactivity of cytoskeleton, resulting in a non-unidirectional modification of cellular function, or immuno-modulation.

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A multicenter study was carried out in patients with RA as a part of the prospective evaluation for safety and efficacy of CCA. A total of 122 patients received CCA for a mean duration of 5.5 months. 69 patients (61.6%) showed better than moderate improvement in the overall improvement evaluation (in evaluated 112 patients) . A statistically significant improvements were noted in the number of painful and swollen joints, joint count, morning stiffness, Lansbury's index and serum IgG. Major adverse reactions were gastrointestinal upset, renal impairment and skin rash. The clinical improvement followed the normalization of peripheral blood lymphocyte count, CD3 and CD4 cells.
Our data confirms that immunomodulating drug CCA is a useful agent in the treatment of patients with RA.

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