In an attempt to study the relationship between response-patterns of the humoral immunity system and types of glomerular pathology in primary renal diseases, B lymyhocyte subsets, serum immunoglobulin levels and glomerular immunopathologic findings were examined. The B lymphocyte subset, which was designated as Br, Ba, Bi, Be or Bo, was identified by the intrinsic surface immunoglobulin on each lymphocyte; IgG, IgA, IgM, IgE or IgD, respectively. In this study we clarified that each primary renal disease had a peculiar B lymphocyte subset pattern as follows: Bε-(Bγ) pattern (pattern with a significant increase in Be and a tendency to increase in Br) in minimal change nephrotic syndrome (MCNS), Br-(Be) pattern in membranous nephropathy (MN), (Br)-(Ba) pattern in mesangial proliferative glomerulonephritis (PGN) and Bγ- (Bα)-(Bμ) pattern in membranoprolif erative glomerulonephritis (MPGN). In MN, PGN and MPGN, the B lymphocyte subset pattern corresponded to the results of immunofluorescent findings of glomeruli observed in each disease. In MCNS, where no specific finding was observed in either light or immunofiuorescent microscopic examination, Be was increased in corresponding to the elevation of serum IgE. These results suggest that the B lymphocyte subset pattern may have an important role in determination of histological type in primary renal diseases.

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Since the introduction of disporsable columns containing polymercoated charcoal, trials of charcoal haemoperfusion have been undertaken in various drug intoxications. We studied the efficiency of direct haemoperfusion (DHP) for the removal of paraquat (1, 1'-dimethyl 4, 4'-bipyridilium dichloride: Gramoxon^R) from circulating solution. In vitro, the concentration of paraquat 5, 10, and 100 ppm in 4 L dialysate were prepared. The flow rate through the column of 100 and 200 ml/min was obtained by the use of pump. In 100 ml/min flow rate, 1-7% of paraquat remained after three hours perfusion. In 200 ml/min flow rate, however, paraquat was not detected in one and a half hours after the start of perfusion (Limit of detection is less than 0.04 ppm). In three years, we experienced 18 cases of herbicidal paraquat intoxications. In 10 of these 18 patients, DHP with hydrocoating charcoal were performed after gastric lavage, forced diuresis, and administration of Fuller's earth. Six of these 10 patients died within 28 days after injestion of Gramoxon quantity varing between 5 to 80 mis. Four of these 10 survive without pulmonary complication and with mild to moderate kidney and/or liver dysfunction during their courses. These complications, however, disappeared complitely within one month. We conclude that charcoal haemoperfusion itself is effective instrument for removing paraquat from solution in vitro, and also for clinical use in some extend. Our experiences, however, indicate that patients who ingested large dose, who were not treated in initial phase, showed poor prognosis inspite of DHP therapy.

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Lysozyme (Ly.) has been ordinary known as anticoagulant agent, antiinflammatory agent and a agent possessing recovering function from the tissue injury. On such pharmacological basis of this drug, we studied the effect of hen-egg Ly. (kindly supplied by Nippn-Shinyaku Co. L.T.D.) on Masugi nephritis and Aminonucleoside nephropathy in rats. Regarding with the effects of Ly. on acute phase of Masugi nephritis, the degree of infiltration of polymorphonuclear leucocytes (PMN) became severe with the increase of the dose. Although fibrin deposition in glomeruli was slight in the groups rate received 60 mg Ly. per rat, there were no distinctive difference between the experimental other groups and control group. Regarding with the effect of Ly. on chronic phase of Masugi nephritis, massive proteinuria was found in all groups without any correlation due to the dosis. On the microscopical findings, all groups of rats with Masugi nephritis showed proliferative and partially hyalinized glomeruli, interstitial mononuclear cell infiltration and degenerative tubular changes. The degree of fibrin deposits seemed to be increased in the groups of rats received Ly., comparing with the control group. Regarding with the effects of Ly. on Aminonucleoside nephropathy, experimental rats developed fairly large amount of proteinuria on 12 th day after Aminouucleoside injection. But no significant difference on the proteinuria was found in all groups of rats. In addition of this, light microscopical findings in all groups similarly showed widening of mesangial area, mild proliferative changes in glomeruli, degenerative tubular changes and interstitial mononuclear cell infiltration, but the degree of fibrin deposition seemed to be severe in the groups of rat received Ly. From these results, the effect of Ly, on the glomerular injury induced by immunological or nonimmunological procedure seemed to have enhancing action against the infiltration of PMN and also coagulation process. Thus, it was suggested that Ly, was of the inductive action for scarring and healing effects of injury site.

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Modification of the original double-isotope radioenzymatic assay of Engelman, K. et al. permits the direct and simultaneous analysis of noradrenalin, adrenalin and dopamine in plasma samples of 100 fcl. This catechol-o-methyltransferase-catalyzed assay is sensitive to 10 pg for noradrenalin, adrenalin and dopamin. Employing this method, plasma noradrenalin and adrenalin were measured in 12 normal subjects and 12 essential hypertention at resting state and during acute stress. In normal subjects lying supine for 30 minutes, plasma noradrenalin and adrenalin levels averaged 176 pg/ml and 71 pg/ml respectively. In essential hypertention, plasma noradrenalin and adrenalin levels were significantly higher, with an average of 240 pg/ml and 86 pg/ml. Insuline-induced hypoglycemia and cold pressor test resulted in a greater increase in plasma catecholamines in essential hypertention than agematched normal subjects. These results suggest that the higher basal values of noradrenalin, adrenalin and the excessive discharge of noradrenalin and adrenalin into plasma during stress may play an important role the development and maintenance of high blood pressure in essential hypertention.

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A 23-year-old woman with idiopathic Fanconi syndrome without cystinosis was described. Her parents were healthy but consanguineous. When she was 9-year-old, the diagnosis of acute glome-rulonephritis was made because of her edema and proteinuria. Addition to proteinuria, renal glycosuria, hypokalemia, and metabolic acidosis were detected next year. Bone pain and arthralgia abrupted when she was 21-year old. She visited to our hospital complaining of arthralgia of hip, knee and ankle joints and bone pain of ribs, then was admitted on July 1978. She was a small lady. Pigeon chest, mild anemia and enlarged spleen were noticed. Laboratory examination revealed mild anemia, hypokalemia, hyperchloremia, hypophosphatemia, hypouricemia, renal glycosuria, proteinuria, increased serum alkaline phosphatase level and urine beta₂-microglobulin excretion and metabolic acidosis. In renal function GFR, %TRP, Tm-PAH, Tm-G were decreased and phosphate clearance, uric acid clearance and filtration fraction were increased. Fishberg concentration test and acidification test of Elkinton modification were impaired. Roentogenological examination of bones represented demineralization and pseudofractures which formulated osteomalacia. Amino acid analysis showed her severe pan-aminoaciduria and mild aminoaciduria of her parents and brother. Cystinosis and another diorders which caused secondary Fanconi syndrome were not disclosed.

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This study was attempted to clarify the effects of ethacrynic acid and chlorothiazide on mitochondrial oxidative phosphorylation. The results show that ethacrynic acid above the concentration 10^-5 M inhibits state 3 respiration and accelerates state 4 respiration of isolated mitochondria from the rat liver, renal cortex and renal madulla. Ethacrynic acid inhibits the respiration uncoupled by pentachlorophenol in isolated mitochondria from the rat liver. Furthermore, ethacrynic acid accelerates the respiration controlled by oligomycin in isolated mitochondria from the rat liver. The activities of NADH-DCIP reductase, NADH-cytochrome c reductase, succinate dehydrogenase and succinate-cytochrome c reductase from the frozen-thawed beef mitochondria are inhibited by the addition of 10^-3 M ethacrynic acid. The inhibition of NADH-cytochrome c reductase is considerably greater than that of NADH-DCIP reductase. These results suggested that ethacrynic acid simultaneously acted as an electron-transport inhibitor and as an uncoupler. This inhibition of mitochondrial electron-transport system could occur at multiple sites. Therefore, mitochondrial oxidative phosphorylation might be inhibited by these complicated biological action of ethacrynic acid. On the other hand, neither the influence upon state 3 respiration nor state 4 respiration is observed by chlorothiazide below the concentration 3.8 × 10^-3 M. Neither the inhibitory action on electron-transport system nor uncoupler action is assumed by this result. The biological properties of chlorothiazide other than mitochondrial fraction such as plasma membrane might be important.

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Fifty nine renal biopsies from 49 children with idiopathic nephrotic syndrome (INS) were studied to see the clinicopathologic and immunologic correlations. There were 27 patients with minimal change lesion (MCL), 8 with mesangial proliferation (MSP) and 14 with focal segmental sclerosis (FSGS) with the average follow-up period of 5, 3 1/2 and 3 years respectively. The presence of focal segmental sclerotic changes, tubular atrophy, interstitial fibrosis and infiltration of mononuclear cells and mesangial hypercellularity has significant correlation to resistance to therapy. However, the presence of mesangial cellularity in FSGS did not correlate with poor clinical response. Most of the patients with MSP showed the steroid resistance, but all had favorable outcome. By immunofluorescent studies segmental patterns were found in FSGS, but fragmentary linear or granular mesangial deposits of immunoglobulins and complements were observed in almost same frequency in MCL and FSGS and less in MSP. The presence of IgM in glomeruli did not correlate with the response to therapy, the serum levels of IgM and the duration of the disease. Measurements of serum immu-noglobulins and complements could not show any difference among three groups of INS. The levels of serum IgM were variable but 50% of the specimens had the serum levels greater than one standard deviation of normal mean. Serum levels of Clq, factor B and C3b inactivator are often decreased in INS. But the meaning of the presence of IgM in glomeruli, elevated serum levels of IgM and decreased levels of Clq, factor B and C3b inactivator is unknown. From these observations immunological mechanisms seems to be involved in INS, although it is still possiblethat these findings could be due to secondary changes, rather than primary. Four patients whose initial biopsy showed MCL but had focal segmental lesions on subsequent biopsy showed distinct steroid resistance from the beginning. From the clinical, pathological and immunofluorescent studies MCL and FSGS seem to be distinct groups. However, the progression of lesion from MCL to FSGS remains unclear.

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The in vitro guinea pig complement fixation test (gpCFT) using immunofluorescent method (IF) on frozen sections of 60 consecutive biopsy cases of various idiopathic glomerulonephritis was carried out of purpose of studying the reserved complement activating ability of regionally immune complexes (IC) in glomeruli The results of this test were compared with the ingredients and intensities of own immunoglobulins and complements depositions in glomeruli and were further compared with the serum complement levels from laboratory data and seventies of light microscopical glome-rular changes. The positive gpCFT was usually found in some of cases which showed positive depositions of own complement components in glomeruli, furthermore positive results of gpCFT were seemed to be well correlated with the high degree of depletion of serum complement levels. On the other hand, although complement components were noted together with certain classes of immunoglobulin forming IC in glomeruli, the positive results of gpCFT were rather scarce in the cases which had IgA in their glomerular deposition compared to the IgA negative counterparts. By morphological study, it was noted that the cases of typical membranoproliferative glomerulo-nephritis and advanced proliferative glomerulonephritis (PGN) showed high rate of positive gpCFT, on the other hand the cases of milder PGN and IgA nephropathy showed rather low rate, despite the fact that the latter cases demonstrated considerable amount of deposition of own complement components to their glomeruli. Hitherto described evidences reveals that positive result of gpCFT could properly suggest the one aspect of reserved biological activity of regional IC, therf ore it might also represent the tempo-rary injurious capacity of IC better than the quantity or intensity of IC deposition per se in glome-ruli of various types of glomerulonephritis.

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12 nephritic patients with proteinuria above 1 g/day were subjected to anticoagulant therapy in combination with pulse therapy. The patients consisted of 6 mesangial proliferative glomerulonephritis, 2 membranoproliferative glomerulonephritis, 2 anaphylactoid purpura nephritis and 2 focal sclerosing glomerulonephritis. In 4 of 6 mesangial proliferative glomerulonephritics proteinuria subsided and renal clearance was improved by treatment with dipyridamole and/or warfarine plus pulse therapy, These 4 patients have been asymptomatic for more than 1 year and a half despite minor hematuria. In one of the other two cases with mesangial proliferation given cyclophosphamide concurrently a complete remission was achieved, but the rest did not respond to the therapy. This form of therapy was without effect besides transient diminution of proteinuria in one of the two cases with membranopro-liferative glomerulonephritis, however, successful outcome including disappearance of proteinuriaa and elevation of serum complement level was obtained in the other. The following 6 patients were treated with combined heparin and pulse therapy. In one of the two anaphylactoid purpura nephritics, who showed massive proteinuria of 15 g/day, this combination therapy was ineffective; only diminution of proteinuria and relief of edema followed but proteinuria of 2-4 g/day is still continuing. The therapy resulted in complete subsidence of proteinuria leading to remission in the other anaphylactoid purpura nephritic with proteinuria of 2-3 g/day. In 2 patients with focal sclerosis a complete remission or a diminution of proteinuria was induced with concurrent use of cyclophosphamide. Thus we consider that the results with this form of treatment are encouraging and warrant further trial in cases refractory to other medications. Heparin is known to be contraindicated to lipoid nephrosis of minimal change group, accordingly, warfarin or dipyridamole is preferable in the management of mesangial proliferative glomerulonephritis. In the patients with membranoproliferative and anaphylactoid purpura nephritis heparin can be used favorable with pulse therapy.

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Unilateral progressive Masugi nephritis was produced in albino rabbits weighing 2.0 to 2.5 kg, according to the method of Sarre and Wirtz modified by Nagasawa. The following four experimental groups were designed. 1) Heparin group (14 rabbits); heparin, 1000 units/kg, twice a day, S. C. was started simultaneously with nephrotoxic serum (NTS) injection, and was given for 3 weeks. 2) Urokinase group (14); this, 1800 units/kg, twice a day, I. V. was started 5 days after NTS and was continued until the sacrifice. 3) Ancrod group (13); this was started 5 days after NTS, and was given, initially, 0.5 units/kg, I.V., and 1 unit/kg, one hour apart, then from the next day, 1 unit/kg, twice a day. 4) Control group (12); no treatment except for NTS. All animals were sacrificed 22 days after NTS. Evaluation was performed as follows. 1) Light microscopic examination is to count the number of glomeruli with circumferential crescent formation per 100 glomeruli. 2) by immunofluorescence using FITC-guinea pig anti-rabbit fibrinogen serum, the degree and number of fluorescent glomeruli were examined per 50 glomeruli. 3) by microangiography according to the method of L junggvist, the number of glomeruli visualized over a certain area of the cortex was calculated. In the control group, the incidence of crescent formation was 59.5±9.0 (SE), while it was 31.9±9.5 (p<0.05) in heparin group, 30.3±8.1 (p<0.02) in urokinase group and 14.8±5.7 (p<0.01) in ancrod group. But, 2 of 13 rabbits given ancrod showed glomerular mesangiolytic injury, known to occur in poisoning by the Habu snake venom. The immunofluorescent studies disclosed a significant increase in the number of negatively stained glomeruli from 3 treated groups. Moreover, the numbers of glomeruli seen per unit of cortex by microangiography in the control, heparin, urokinase and ancrod groups were 70.2±6.5, 104.3±6.0 (p<0.001), 99.0±7.4 (p<0.01), 102.4±7.2 (p<0.01), respectively. The present result demonstrates that three kinds of agents including ancrod are equally effective in spite of different mechanisms of action for attenuation of intraglomerular coagulation induced by the experimental nephritis. But, the mesangiolytic injuly observed in a few rabbits given ancrod suggests its need to be more purified for future clinical use.

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A study was performed in order to elucidate the metabolic change of ³⁵S-furosemide (³⁵S-FM) in control (C), renal failure (RF) and bilaterally nephrectomized rats (Nx). The suppressing effects of probenecid on ³⁵S recovery and FM metabolites in urine and bile were also studied. Serum ³⁵S-FM concentration in RF was higher than that in C, and T1/2 was 28 min in C and 34 min in RF. ³⁵S recovery in urine within 2 hr following ³⁵S-FM administration was 57% in C and 32% in RF, whereas ³⁵S recovery in bile was 20% in C and 25% in RF. Fifty three percent of ³⁵S was excreted into bile in Nx. Thus, ³⁵S excretion into bile increased in RF and Nx. The FM metabolites, 4-Chloro-5-sulfamoyl anthranilic acid (CSA), Rf 0.76, and another metabolite (P-3), RI 0.50 were separated by thin layer chromatography and infrared spectrophotometry. FM and its metabolites were discovered in urine, bile and serum in C and RF. With respect to FM metabolism in C, the unmetabolized fraction of FM in urine was 68%, whereas that of FM in bile was only 10%. The fraction of unmetabolized FM increased in RF. FM metabolites disappeared from urine and decreased in bile following administration of probenecid. It is concluded that there is compensatory mechanism in biliary FM excretion in RF. Two substances are identified as FM metabolites which have no significant diuretic activity. Probenecid inhibits the excretion of FM and its metabolites.

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