Several mediators released from the synovial cells, synovial lymphocytes, and polymorphonuclear leukocytes (PMN) or lymphocytes in synovial fluids of rheumatoid patients were overviewed and the function of the major mediators as proteinase, IL-6 were demonstrated.
In addition, a couple of experiments done in our laboratory were demonstrated. Synovial fluid PMN adhered to the cultured synovial cells and damaged them in both morphologically and making a release of ⁵¹Cr from the labelled-synovial cells. A part of the damaging mediators was sensitive to catalase and trypsin inhibitor.
On the other hand, synovial fluid lymphocytes stimulated the synovial cells in showing a colony formation of the synovial cells and also in increasing the ³H-thymidine up take. A part of the stimulating mediators seemed to be interleukin 2. Synovial fluid lymphocytes secreted also factors which could enhance the damaging effects of synovial fluid PMN and a part of the factors was identified as interferon γ.

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Intravenous (i.v) injection of human recombinant interleukin-1α (IL-1α) or interleukin-1β (IL-1β) produced significant increases in the deep body temperature and in the plasma concentration of adrenocorticotropic hormone (ACTH) in rats.
In the present study, it was investigated whether intrahypothalamic prostaglandins (PGs) are involved in the development of the ACTH response which is induced by IL-1's. The pre-treatment with a cyclo-oxygenase inhibitor, indomethacin, inhibited completely the febrile responses and attenuated the increases in the plasma concentrations of ACTH induced by the i.v injection of IL-1α or IL-1β, indicating that the enhancement of plasma levels of ACTH induced by the i.v injection of IL-1's is processed through the action of prostaglandins. Intrapreoptic injection of PGE₂ produced a monophasic fever with a rapid onset. Moreover, intrapreoptic injection of PGE₂ made significant increases in the plasma concentration of ACTH 30 min after injection.
These results suggest that intrapreoptic PGE plays an important role in the ACTH response by inducing the release of corticotropin releasing factor.

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We have developed a mouse marrow culture system in which multinucleated osteoclast (OC) -like cells are formed within 8 days. Using this culture system, we examined the effect of prostaglandins (PGs), potent bone-resorting agents, on OC-like cell formation. Four PGs (PGE₁ and PGE₂ at 10^-8-10^-5 M, 6-keto-PGF_1αat 10^-5M, and PGF_2αat 10^-6-10_-5M) significantly stimulated the formation of OC-like cells. The potency of the PGs in inducing OC-like cell forma-tion was well correlated with the potency in increasing the production of cyclic adenosine 3', 5'-monophosphate (cAMP) in bone marrow cells. Addition of dibutyryl-CAMP also induced OC-like cell formation. Moreover, isobutylmethylxanthine (IBMX), a potent inhibitor of phosphodiesterase, potentiated the OC-like cell formation induced by PGE₂. Calcitonin induced cAMP production in cultures treated with PGE₂, but not in cultures with vehicle. When bone marrow mononuclear cells were cultured on dentine slices in the presence of PGE₂, multinucleated OC-like cells were similarly formed and they resorbed calcified tissues.
These results suggest that PGs stimulate resorption of calcified tissues by promoting osteoclast formation. The activity of PGs in inducing OC-like cell formation is considered mediated by a mechanism involving cAMP.

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On inflammatory initiation, macrophages are enhanced in their capacity to produce reactive oxygen intermediates such as O^-₂ and H₂O₂, and kill microbes for host survival from infection. Contrary to the initiation, the mechanism to cease inflammation is hardly understood. Since permanent producing of reactive oxygen intermediates is toxic, the host itself probably requires the ability to deactivate macrophages. Macrophages are essential to the healing of wounds and repair of tissue damaged by inflammation. These concepts led us to search for macrophage deactivating effects among polypeptide growth factors that regulate angiogenesis, fibrogenesis and other aspects of tissue repair. Among 12 such factors, two polypeptides that possess 71% similar homology proved to be potent macrophage deactivators: transforming growth factor-β1 (TGF-β1) and TGF-β2.
The fact that their concentrations for 50% inhibition were extremely low of picomolar level, suggests TGF-β play a powerful role on ceasing the inflammation and tissue repair after an inflammation site has been sterilized. Although the deactivating site by TGF-β existed in anabolism but nct catabolism of reactive oxygen intermediates, exact site is not yet determined.

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Human alveolar macrophages (PAM) play an important role in host defence in the lung by secreting several biologically active molecules including eicosanoids to elicit inflammatory reactions. We investigated the effect of smoking and pretreatment with lipopolysaccharide (LPS) on the secretion of leukotrienes (LT) and prostaglandins (PG) from PAM. PAM were recovered by bronchoalveolar lavage, and cultured in vitro with or without LPS (500 ng/ml) . After various periods of incubation, PAM were washed and then stimulated with calcium ionophore A23187 (2μM), or phorbol myristate acetate (PMA, 500 ng/ml) . LT and PG in supernatants were quantitated by high performance liquid chromatography. Minimal detectable amount of LT and PG were 1ng. LTB₄ was detected in the supernatant of PAM stimulated with A23187 as a sole LT, and PGE₂ in the supernatant of nonsmokers' PAM pretreated with LPS and then stimulated with PMA as a sole PG in this assay system.
Smokers' PAM secreted decreased amount of LTB₄ compared with nonsmokers' PAM. No PG was detected in the supernatant of smokers' PAM even if they were pretreated with LPS. PAM pretreated with LPS secreted increased amount of LTB₄ compared with control. Thirty minutes preincubation was optimal for PAM to secrete LTB₄. In contrast, twelve-hour pretreatment was required for nonsmokers' PAM to secrete detectable amount of PGE₂, and the secretion of PGE₂ increased up to 48-hour pretreatment.
In conclusion, smoking decreased the capacity of PAM to secrete LTB₄ and PGE₂, and LPS primed PAM for enhanced secretion of LTB₄ and PGE₂.

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Medullasin is a neutral proteinase first purified by Aoki from the mast cells of bone marrow (Aoki (1978) J. Biol. Chem. 253, 2026-2032) . It causes an artificial inflammation when injected into the skin of experimental animals, thus considered to be an agent responsible for the occurrence of pathogenic inflammation. The proteolytic activity of medullasin was effectively inhibited by stoichiometric amounts of a serum proteinase inhibitor α₂-macroglobulin and its egg white homolog, ovomacroglobulin.
Competitive inhibition of medullasin by α-1-proteinase inhibitor and α₂-macroglobulin was studied under a simulated in vivo condition and it was shown that even in the presence of 30 fold molar excess of α-1-proteinase inhibitor 30-40% of medullasin was bound to α₂-macroglobulin, indicating that the latter inhibitor can be an effective modulator of inflammation in vivo.
Ovomacroglobulin was also proved to be an effective inhibitor under similar conditions.

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Electron spin resonance (ESR) is regarded as the least ambiguous method for the detection of free radicals. Using spin trapping technique with 5, 5-dimethyl-1-pyrroline-N-oxide (DMPO), we measured superoxide anion radicals generated by human polymorphonuclear leukocytes (PMN) stimulated with various agents. PMN were isolated from heparinized venous blood of healthy volunteers by dextran sedimentation followed by Ficoll-Paque separation and hypotonic lysis of contaminating erythrocytes, and suspended in Hanks' balanced salt solution at pH 7.4. Phorbol myristate acetate (PMA), opsonized zymosan (OZ), digitonin, and Ca ionophore (A23187) were used as stimulants of PMN. ESR spectra were obtained by JEOL-JES-FR80 ESR spectrometer, and the intensity of the signal of DMPO-OOH and DMPO-OH adduct was measured as the ratio to the intensity of Mn²⁺ signal.
The ESR spectra produced by stimulated PMN in the presence of DMPO was composed of DMPO-OOH adduct and DMPO-OH adduct, however, the composite ratio of signal intensity of DMPO-OH adduct to that of DMPO-OOH addutt differed with stimulants of PMN. The relative intensity of DMPO-OOH signal increased in proportion to the cell concentration, indicating that semi-quantitative determination of superoxide generated by stimulated PMN is possible.

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We performed bronchoalveolar lavage (BAL) in 30 HTLV-1 carriers. They include 17 HTLV-1 associated myelopathy (HAM) patients with chest X-ray abnormality (8 cases) and without chest X-ray abnormality (9 cases) and 13 non-HAM HTLV-1 carriers with pulmonary involvement. We then investigated the immunologic aspects of the cells obtained from BAL. Increased % lymphocyte was observed in 60% and increased neutrophil was observed in 50% of the patients respectively. Lymphocyte surface analysis used monoclonal antibodies revealed that increased lymphocyte were T-lymphocytes.
It seems that CD4⁺T lymphocyte appears to be dominant in the case with manifest chest X-ray abnormalities. HTLV-1 antigen positive lymphocytes were detected by indirect immunofluorescent method used monoclonal antibody, GIN-14 (anti-p19) and F-10 (anti-gp21) in the BAL cell culture. Moreover significant high immune responsiveness against HTLV-1 was observed in BAL cells obtained from HAM patients with chest X-ray abnormalities. These observations suggest that the pulmonary involvement in HTLV-1 carrier may relate to HTLV-1 infection and immune responsiveness against HTLV-1 in the lung.

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The present clinical trial was attempted to compare and evaluate the degree of usefulness of flurbiprofen adhesive topical transdermal delivery system (FP-A) as a non-steroidal anti-inflammatory analgesic that reduces systemic side effects, enhanced drug concentration within the tissues of the local inflammatory site beside eliciting a persistent relieving effect on the affected site. The trial was performed under double blind comparative study with subjects suffering from periarthritis scapulohumeralis and where the FP-A base was used as the control. The following results were obtained:
(1) Of the total of 191 cases with periarthritis scapulohumeralis, 141 achieved general consequent improvement, overall safety index for 182 cases, and 147 cases had degree of usefulness on analysis of the subjects studied.
(2) In cases where general consequent improvement was seen, a excellent improvement was noted in 35.6% (26/73 cases) when compared to FP-A base group of 16.2 % (11/68 cases) with the former group showing a significant (P<0.05) improvement. For cases with above “good improvement”, improvement was seen in 64.4% (47/73 cases) in the FP-A goup whereas 42.6% (29/68 cases) in the FP-A base group, indicating the former to register a significant (P<0.05) improvement over the latter.
(3) With regards to the overall safety index, cases that were judged to “have problems in safety margin” included 2.2% (2/89 cases) and 4.3% (4/49 cases) in the FP-A and FP-A base groups, respectively. Moreover, for cases that wore categorized as “not safe” in the FP-A group, a only 1.1% (1/93 cases) was observed.
Side effects were seen confined to the sites where the adhesive was patched. Appearance of these side effects and safety margin was not at all significant between the 2 groups.
(4) On the degree of usefulness, cases that categorized as above “extremely useful” were 34.2% (26/76 cases) in the FP-A group against 15.5% (11/71 cases) in the FP-A base group. For those that were above “good useful”, 63.2% for the former and 42.3% for the latter were noted. For the 2 categories above, treatment given to the FP-A group was significantly (P<0.05) beneficial when compared to the FP-A base group.
The above findings reveal FP-A to elicit a clinically beneficial effect on, and therefore is a beneficial medicament for, periarthritis scapulohumeralis.

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