There is a considerable body of evidence that inflammatory response is mediated by various cytokines. IL-5 is one of inflammatory cytokines. The study of IL-5 has been originated in the search for one of the B cell growth and differentiation factors, named T cell-replacing factor (TRF), that induces antigen-stimulated B cells to differentiate into plasma cells. Eosinophil differentiation factor (EDF) has been known as a diffusible factor from thoracic duct lymphocytes of parasite-infected rats. The cDNA cloning and mAb against IL-5 enable us to identify this molecule as a cytokine (TRF/EDF) that has pleiotropic activity in vitro on various target cells besides B cells and eosinophils. The IL-5 transgenic mice show not only polyclonal B cell activation represented by enhanced serum levels of IgM, IgA, and IgE and autoantibody production, but also massive eosinophilia in peripheral blood and eosinophil infiltration into various tissues.
The pleiotropic activity of IL-5 on target cells is directly dependent on initial binding to specific cell-surface receptor (IL-5R) . IL-5 interacts with target cells with biphasic equilibrium binding kinetics, reflecting two classes of binding sites with high and low affinity. We carried out the cloning of the cDNA for the IL-5Rα chain which found IL-5 with low affinity by itself. The IL-5Rα chain associates with the β chain which can convert low affinity IL-5R to the high affinity. The β chain of IL-5R, that does not bind either IL-5, IL-3, or GM-CSF by itself is shared among IL-5R, IL-3R, and GM-CSFR.
These results can imply why IL-5, IL-3, and GM-CSF are eosinophil-poietin, but can not explain why IL-5 preferentially induces eosinophil production. IL-5 specific signalling may be transduced through IL-5Rα chain. We need further experimentation to give definite answer on these issues.

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Platelet-derived growth factor (PDGF) -A and -B gene expression was studied using resting monocytes, in vitromatured monocytes and alveolar macrophages. Resting monocytes constitutively transcribed both PDGF-A and -B genes. When monocytes were matured to macrophagesin vitro, the transcription rates of both genes increased markedly. Consistent with the transcription rates, resting monocytes constitutively expressed both PDGF-A and -B mRNAs. Interestingly, PDGF-B mRNA levels increased markedly after the maturation, whereas PDGF-A mRNA levels did not change. Alveolar macrophages constitutively transcribed both PDGF-A and -B genes at almost the same rates. However, these cells expressed 5-fold more PDGF-B mRNA than PDGF-A mRNA. Immunohistochemical study using anti-PDGF-AA and anti-PDGF-BB antibodies suggested that PDGF-BB homodimers were more abundant than -AA homodimers or -AB heterodimers in alveolar macrophages and in vitromatured monocytes.
These observations indicate that PDGF-B mRNA is preferentially expressed and PDGF-BB homodimers are abundantly produced after the maturation of monocytes to macrophages, despite the equal transcription rates for both PDGF-A and -B genes.

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Important roles of reactive oxygen species (ROS) to microbicidal functions of neutrophils are well recognized. Recently, chemiluminescent techniques, that have been used in research fields of oxygen radicals, are becoming to be much easier by technological advancements in this decade. Many investigators have reported using chemiluminescent techniques the ROS generation from phagocytes. But unfortunately, methods to evaluate the ROS generating ability of neutrophils from chemiluminescent measurements have not been well established yet.
In this study, to establish a new estimation method for the neutrophil activity, a kinetic approach is proposed and parameters of reported estimation methods are reconsidered by comparison with an obtained ideal equation by the kinetic approach.
Applications of the ideal equation to actually measured values using Zymosan, E. coli, andS. aureus as particles showed good concurrences. By the comparison between reported esitmation methods and the obtained ideal equation, following results are observed. 1) Initial slopes (tanΘ) of a slope method may have a possibility to estimate a neutrophil activating constant, ka, of the ideal equation. 2) Peak heights of a peak method and under curve areas of a area method are linearly correlated with a statistical significance. But limitations of the area method to estimate the neutrophil activity are revealed by the ideal equation. 3) For the peak method, it is possible to estimate the neutrophil activity, if both parameters of the peask method, peak heights and peak time, are well considered.
The results of this study contribute much to the extension of knowledge of the neutrophil chemiluminescence.

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Effect of acidic non-steroidal anti-inflammatory drugs (NSAIDs), indomethacin, floctafenine and tiaprofenic acid, on active oxygen generation from guinea pig peritoneal (GPtPMN) and human peripheral neutrophils (HPPMN) were studied. NSAIDs suppressed formylmethionyl-leucyl-phenylalanine (FM LP) -induced active oxygen generation from GPtPMN.
Granulocyte colony-stimulating factor (G-CSF) enhanced receptor-mediated O^-₂ generation. Such a priming effect was closely correlated with proteintyrosyl phosphorylation of specific HPPMN proteins. NSAIDs also suppressed both the priming effect and the protein-tyrosyl phosphorylation by G-CSF in concentration dependent manners, and the effective concentrations were lower than those for the active oxigen generation from GPtPMN.
These results suggest that NSAIDs effect not only an activation step (s) of NADPH-oxidase but also an early step (s) of neutrophils activation.

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Formation of subcorneal pustules characterizes the skin lesions infected byStaphylococcus aureus (S. aureus) . To elucidate the mechanism underlying the pustule formation as well as that of anti-bacterial host defence, we studied the effect of protein A on the interaction between corneocytes and neutrophils.
We found that protein A significantly promoted opsonized corneocytes-induced chemiluminescence (CL) in neutrophils. It took place even with agammaglobulinemia serum, ruling out the possibility of the involvement of Fc-receptors of neutrophils. Microscopic observation of such corneocytes revealed an increase in the number of neutrophils adhering to their surface. Ultrastructural observation showed a distinct deformation of the neutrophils adhering the surface of the corneocytes. Such features indicating enhanced frustrated phagocytosis of neutrophils seem to suggest that they play an important role in the host defence mechanisms against the invadingS. aureusby producing the characteristic pustules due to the neutrophil-mediated damage of the surrounding tissue.

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The auther studied a role of leukocyte-elastase in vascular endothelial cell injury induced by activated leukocytes. Using an interactive laser cytometer with ACAS 570, the intracellular peroxide level was determined in dichlorofluoresein-diacetate (DCFDA) prelabeled vascular endothelial cells. Addition of elastase (5 μg/ml) to the monolayer of the prelabeled endothelial cells induced an increase in fluorescence intensity in cells. The increase in fluorescence was inhibited by such protease inhibitors as α-1 antitry-psin and α-2 macroglobulin. Moreover, pretreatment of the endothelial cells with allopurinol, xanthine oxidase inhibitor, abolished the increase in the fluorescence of the DCFDA prelabeled endothelial cells exposed to elastase.
These data suggest that the activity of leukocyte elastase is related to the production of peroxide in endothelial cells. Elastase may activate xanthine oxidase or increase the conversion of xanthine dehydrogenase to xanthine oxidase, and it may be deeply involved in vascular endothelial cell injury elicited by activated leukocytes.

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Enlargement of the seminal vesicles (SV) and subjective pain are very important factors for the diagnosis of seminal vesiculitis. The relationship between enlargement of SV and pain was studied experimentally.
A total of 25 male Wistar rats weighing between 300 and 400 g were divided into 5 groups: control; saline injection; and 3 saline injection after pretreatment groups, consisting of intraluminal HCl washing of SV (chemical stimulation), compression with forceps of SV (physical stimulation) and intraperitoneal Aspirin DL-lysine injection, as the pretreatments. For the saline injection groups physiological saline was injected into SV at the speed of 1 and 2 ml/min through a tube placed in SV through a subcutaneous tunnel. Expression of pain was judged by an original criteria including twisting and turning of trunk and so on. No pain was observed in control group and group with analgesia. Pain was expressed in other 3 groups and was specially exaggerated in chemical and physical stimulation groups.
Thus the relationship between enlargement of SV and pain was proved.

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Monocrotaline (MCT) causes lung inflammatory responses and chronic pulmonary hypertension in rats. Since platelet-activating factor (PAF), a potent mediator of inflammation, increases in the lung tissue of MCT-treated rats, we examined the effect of WEB 2170, a specific PAF-antagonist, on lung in-flammatory responses and subsequent pulmonary hypertension induced by MCT. Treatment with WEB 2170 significantly reduced pulmonary hypertension and right ventricular hypertrophy at 3 weeks after MCT-injection. The eicosanoid levels in the lung tissue and the number of abnormal alveolar macrophages increased in MCT-treated rats at 3 weeks. In WEB 2170-treated MCT rats, the lung 6-keto prostaglandin F_1αlevels and the number of abnormal alveolar macrophages were significantly lower than those of MCT-treated rats.
Theae results indicate that the PAF-antagonist inhibits the development of pulmonary hypertension induced by MCT, and suggest a role for PAF in the lung inflammatory process that contributes to the development of pulmonary hypertension of MCT-treated rats.

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The antifungal and antibacterial activity of both anti-ulcer terpens and sofalconein vitrowere studied.
Among them, plaunotol has proved to show the strongest antibacterial and antifungal activity. It exhibits a wide spectrum activity against dermatophytes and Gram positive bacterias. The weakin vitroactivity of sofalcone against dermatophytes and Gram positive bacterias was recognized and also gefarnate revealed the clinically useful MIC values, whereas teprenone didn't indicate antifungal and antibacterial activityin vitro.
Mechanism of fungicidal action is not clear, but present study suggests that surplus C₂₀unit inhibits the transport of several glycoprotein of cytoplasm and/or metabolites of plaunotol having aldehyde may inhibit the growth of various dermaphytes and bacterias. These results indicate that anti-ulcer terpens and sofalcone may be useful for the treatment of superficial fungal infections.

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Mechanisms how anti-rheumatic agents work in patients with rheumatoid arthritis have not been well clarified. We determined urinary excretion of bucillamine (SA 96) and its methylated metabolite (SA 679) in patients with rheumatoid arthritis, and attempted to find the relationship between the metabolism of bucillamine and its clinical effectiveness.
After taking 100 mg bucillamine orally, SA 96 and SA 679 were excreted into urine 0.3 to 26.0 mg, and 2.7 to 32.3 mg, respectively in 4 hours. Thus, the mode of excretion of bucillamine and its metabolite showed great heterogeneity in patients with rheumatoid arthritis. This was the case with SA 679/SA 96 ratio which represented methylation rate of bucillamine. There was no association between the excretion of SA 96 and SA 679 with age, disease duration, renal function, and total dosage of administered bucillamine.
No significant association was found between the mode of urinary excretion of SA 96 and SA 679 and clinical responsiveness, but some patients responded effectively to bucillamine even though relatively low urinary excretion of SA 96 and SA 679.
In summary, individual differences in the extent of methylation of bucillamine were present in patients with rheumatoid arthritis, however, are unlikely to be directly related to the clinical responsiveness of this anti-rheumatic drug.

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In the treatment of RA, if only rubbing a drug into the inflammation area makes it possible to suppress the symptoms instead of systemic dosing of steroids, such external steroid will be probably useful. Because it doesn't produce side effects due to systemic dosing of steroids.
PNF-21 is an external steroid percutaneously absorbed that has been developed from the viewpoint above mentioned.
The therapeutic effects of PNF-21 were compared with those of Mobilat in 189 patients with rheumatoid arthritis. Both drugs formulations are rubbed into affected area in 3-4 times per day, and applied for 4 weeks.
The results of this study are summarized as follows :
1) PNF-21 showed significant superiority to Mobilat in“final improvement rating”. Improvement rating was 50.0% (40/80) for PNF-21 and 27.4% (23/84) for Mobilat in terms of“moderate”plus“marked”improvement category.
2) The incidence of an adverse reaction was noted as 1.1% (1/92) of PNF-21 and 3.1% (3/96) of Mobilat. But, the abnormal laboratory finding was not observed.
3) The item evaluated, “global utility rating”showed significant superiority of PNF-21 to Mobilat. The utility rating was calculated as 48.8% (39/80) of PNF-21 and 27.1% (23/85) of Mobilat in terms of“moderately”plus“markedly”useful category.
4) Side effects were observed in patient (1.1%) of PNF-21 and 3 patients (3.1%) of Mobilat. All of these symptoms disappeared by symptomatic treatment or discontinued test drugs.
All the above results indicate that PNF-21 is more useful than Mobilat. Therefore, PNF-21 is judged as a highly useful anti-inflammatory agent for the treatment of rheu-matoid arthritis.

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Clinical efficacy of low-dose content tablet Buccilamine (Rimatil (R) 50, Bc 50) was evaluated in 58 patients with active rheumatoid arthritis (RA), 24 of whom had ever experi-enced adverse reactions against regular-dose tablet Bucillamine (Rimatil (R) 100, Bc 100) . Most of the remaining 34 patients had either demonstrated untoward side-effects against or shown no beneficial response to other kinds of anti-rheumatic drugs (DMARDs) .
Initial mean dose of Bucillamine was 59.1mg per day; the mean time span of the treatment with this drug was 23.7 weeks.
At the 16th week of this clinical study, 48% of the patients showed moderate or marked improvement and about 88% of them demonstrated no side-effects, while at the end of this investigation, moderate or marked improvement was observed in 60% and 21% experienced adverse reactions. Clinical parameters, such as duration of morning stiffness, ESR values, grip strength, number of swollen or tender joints and Lansbury's activity index tended to show very favorable move after four to eight weeks of the treatment. Of those 24 patients who had a history of adverse reactions to higher dose of Bucillamine (Bc 100), 17 remained free from side-effects in this study, while more than half (53%) of those who ever demonstrated untoward reactions to other kinds of DMARDs developed side-effects with this low-dose of Bucillamine.
Although the final global utility rate of this drug was somewhat low (46.6%) among this group of RA patients, the figure is fairly reasonable one, considering the lower content of active ingredient cotained in the tablet. Bc 50 tablet is considered quite useful for instituting more flexible dosage set-up in RA patients, including those who ever developed undesirable side-effects against higher dose of this drug, Bc 100.

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A new non-steroidal anti-inflammatory drug, CS-670 was given orally in varying doses to healthy male volunteers for the study of safety and pharmacokinetics.
The following results were obtained:
1) After single (5, 15, 30, 60 and 120mg), divided (180mg/dose, 3 times a day), and multiple (180mg/dose, 3 times a day for 7 days) administrations, no abnormal clinical signs and laboratory examination values attributable to the drug were observed.
2) CS-670 and its four major metabolites, trans-OH, cis-OH, unsaturated-OH, and saturated keton, in plasma or urine specimens were separated using Bone Elut^®C8 column and quantified with high-performance liquid chromatography.
3) The pharmacokinetic parameters didn't change between administration before meal and after meal.
4) After multiple administration of CS-670 three times a day for seven days, plasma concentration and urinary excretion of CS-670 and its metabolites were not significantly different from single administration. Plasma levels of CS-670 and its metabolites reached a steady state at first day.

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