Chemicals emitting light in visible region, when they are oxidized at physiological pH, have been used for testing the generation of active oxygen species and related oxidants. Of these chemicals, luminol has been widely used for testing the function of phagocytizing monocytes and granulocytes. Luminol is known to be oxidized to aminophthalic dianion which in turn emits a light with maximum at 425 nm in aqueous media. The various oxidants generated during phagoctizing, such as O^-₂, ^-O Cl and myeloperoxidase compound III, are involved in luminol dependent luminescences.
Lucigenin, also a chemiluminescence compound, emits a light in visible region during phagocytizing, but produce much move weaker light compared with luminol in phagocytizing granulocytes. Lucigenin-dependent luminescence, however, participates in the generation of O^-₂ (probably also H₂O₂) but not in the myeloperoxidase-mediated reactions.
Very sensitive and specific method for testing an ability of O^- generation in phagocitizing granulocytes and macrophages is considered to be a chemilunescence probe with a crypridina luciferin analog. The compound is, in the main, oxidized to an excited carbonyl in singlet state by O^-, but not by H₂O₂. Under specified conditions, O generating ability of macrophages and granulocytes can be calculated and expressed as xanthine oxidase units.

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Phospholipids and polyphosphoinositides were separated by one-dimensional thin-layer chromatography (TLC) on silica gel plates and lipid containing areas were identified by staining with iodine. Difficulties in separating phosphatidylserine and phosphatidylinositol were encountered in the TLC system on silica gel G-25 plates. Among several plates tested good separation of each phospholipid was obtained in the TLC system of a 5% (w/v) (NH₄) ₂ SO₄ impregnated silica gel H plate developed with chloroform/methanol/acetic acid/water (50: 25: 8: 1, v/v/v/v) at room temperature. Triphosphoinositide, which is one of polyphosphoinositides, did not migrate on silica gel G plates with several solvent mixture. Polyphosphoinositides were well resolved from the other phospholipids in the following TLC system: a silica gel G-25 plate developed with 1% (w/v) aqueous solution of oxalic acid were dried at room temperature, actiated at 110°C for 15 min and developed with chloroform/methanol/4N NH₄OH (9: 7: 2, v/v/v) after loading the lipids.

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SRS-A immunologically formed in rabbit conjunctiva by egg albumin was estimated by HPLC. After the incubation of inflamed conjunctiva with C¹⁴-Arachidonic acid, radioactivity was seen to be incorporated into the SRS-A fraction (Leukotriene C₄, D₄ and E₄) .
Leukotriene C₄ release into medium from rabbit conjunctiva incubated in Tyrode's solution with indomethacin was estimated by radioimmunoassay technique.

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Plasma exchange was performed in a total 18 patients of 11 rheumatoid arthritis (RA), 3 progressive systemic sclerosis (PSS), 1 polymyositis (PM), 1 dermatomyositis (DM) and 2 mixed connective tissue disease (MCTD) . Serum levels of immunoglobulins and complement and also numbers of lymphocyte subsets and its function were measure before and after treatment. Plasma separator AP-05-H (ASAHI Medical Co.) with menbrane filters was used as substitute for the exchanged plasma of 2, 000 to 3, 000ml in each trial.
The treatment seemed to be effective in 9 of 11 RA patients and two of which showed a disappearance of their subcutaneous nodules. The Skin ulcers in the legs found in 2 PSS patients decreased in their size and sclerosis of the skin found in one PSS patient relieved. Respiratory disorder found in one PM patient improved. Muscle weakness and histologic finding of myositis found in one DM patient improved. Severe myalgia and muscle weakness in one MCTD patient improved.
In laboratory data, serum levels of total protein, γ-globulin, immunoglobulin class of IgG, IgA and IgM, C3 and C4 decreased significantly. Decrease of numbers of OKT-8 positive cells and increase of the ratio of OKT-4/OKT-8 positive cell were significant.

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Disodium 4-chloro-2, 2'-iminidibenzoate (CCA) is a drug developed in Japan, and its significant efficacy in rheumatoid arthritis has been demonstrated in multicenter double-blind controlled trials. Since CCA is devoid of direct anti-inflammatory activity, it is suggested that the therapeutic effect of CCA in patients with rheumatoid arthritis may be attributable to improvement of immunological abnormalities that underlie this disease. We investigated changes of lymphocyte subsets in patients undergoing CCA therapy, by assays using OKT3, OKT4, OKT8 and OKIa1 monoclonal antibodies. The T 4 cell subset had no change in blood but showed a tendency to decline in synovial fluid. A significant increase of T 8 cells occurred in blood following CCA therapy while this T cell isubset remained unchanged in synovial fluid. Ia cells decreased significantly in blood whereas they tended to increase in synovial fluid. The drug exerts pharmacological actions that suit the purpose of treatment by way of modulating immunological abnormalities in rheumatoid arthritis.

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In order to elucidate the action mechanism of natural products including Chinese herbs and natural foods which are now widely used for preserving health and/or medical treatments, the effect of sixteen Chinese herbs and two natural foods (AOA, GM) on active oxygen (AO) generated by human neutrophils and in xanthine-xanthine oxidase system was examined. The effects of non-heated or heated agents without dialysis and those after dialysis (which is able to separate molecular weight above 10, 000 from that below 10, 000) on AO levels were compared.
These heated agents markedly and non-heated ones moderately or slightly decreased AO levels generated in both neutrophils and xanthine-xanthine oxidase system. Of interest is that paradoxical findings were obtained from the present experiments that the capacity of these agents for reducing AO levels was not attenuated both by heat and after dialysis, seemingly suggesting that neither superoxide dismutase (SOD) nor low weight chemicals play a role in quenching AO in these agents. However, the capacity of heated ones to decrease AO was significantly attenuated after dialysis. From these results obtained, it seems likely that in non-heated products, AO quenching effect is due to high molecular weight chemicals, enzymes and that in heated ones, this effect is ascribable to the low molecular weight chemicals which are isolated to be activated for quenching AO levels by heat which may have cleaved the bonds of these chemicals linking to other chemicals in natural products. It may be possible that before heated, polymerization of these effective, low molecular weight chemicats to other chemicals inactivates their capacity for removing AO. It also seems to be due to the bondscutting effect of heat with resultant passage of the free low molecular weight, effective chemicals through the diaphromembrane that decreased AO scavenging effect of the products by dialysis after heat. It can be further suggested that these natural products tested have potent AO removing effects and that not only high molecular weight chemicals, enzymes such as SOD but also low molecular weight ones such as flavonoid which have various pharmaceutical action mechnaisms other than scavenging AO may be effectively contained in these natural products.

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