Chronic serum sickness nephritis could be induced with high frequency by multiple preimmunization and subsequent daily intraperitoneal injection of 50 mg/kg of BSA in C57BL/B10-BR mice. Compared with those mice with 2 and 3 times' preimmunization, the mice with 4 times' preimmunization had high affinity of anti-BSA antibody and a larger amount of intermediate size of immune complexes compatible with 19S or smaller, although antibody titers just before starting of daily injection were equally high among those mice with 2, 3 and 4 times' preimmunization. After daily injection of BSA, markedly proliferative glomerulonephritis developed in 95% of 4 times preimmunized mice, while in 50% of 2 times and in 55% of 3 times preimmunized mice. 2 and 4 weeks after starting of daily injection, anti-BSA antibody titers, amounts of circulating immune complexes and levels of free BSA in sera were significantly lower in nephritic group of all mice with 2, 3 and 4 times' preimmunization than in non-nephritic group. In the sera obtained from nephritic mice at 4 weeks after starting of daily injection, amounts of intermediate size of immune complexes apparently decreased and amounts of smaller size of immune complexes compatible with 7S fraction remained unchanged, compared with a chromatographic pattern of immune complexes formed in vitro before daily injection. These findings indicate that according to the increase of times of preimmunization, affinity of anti-BSA antibody becomes higher and a large amount of intermediate size of immune complexes may be formed after daily injection of BSA, and that these immune complexes may be facilitated to deposit in glomeruli, resulting in development of proliferative glomerulonephritis with high frequency.

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In the renal tissues from 16 patients with primary IgA nephropaty, localization of immunoglobulins (IgA & IgG) and complement components (C3, C4, C5 & C9) were studied by immunoelectron microscopy, and the association with Electron Dense Deposit (E.D.D.) was also investigated. IgA was predominantly detected beneath the basement membrane of mesangial area. IgG was present in about half of the patients. Its distribution pattern was similar to that of IgA. Among complement components, the rate of positivity was lower in C4 than in the other late components, C3, C5 and C9, which were frequently detected in the area beneath the mesangial basement membrane. Deposits of C3, C5 and C9 were found in the subendotherial and intramembranous areas. Only C9 was stained in the subepithelial area. These findings suggest that in IgA nephropathy, the complement system is activated mainly by the alternative pathway and also partly by the classical pathway. Although immunoglobulins and complement components were commonly present in E. D. D., these were also detected independently. Some E. D. D. were not stained with immunoglobulins or complement components. This observation raise a question as to whether E. D. D. is always related to the immune complexes.

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Three inbred strains of mice, BALB/c, C3H/He, and C57BL/6 were immunized with allogenic tubular basement membrane (TBM) antigen purified from outbred ddY mice. Renal histological changes of these mice were examined by photomicrograph and direct immunofluorescence. Immune response to IBM antigen was studied by quantification of anti-TBM antibody and proliferative response of nylon wool r cells to TBM antigen. Remarkable differences were obtained in the occurrence of interstitial nephritis (IN) among three inbred strains. BALB/c mice were highly susceptible to IN, whereas C57BL/6 were not. Immune response to TBM antigen, antibody and/or proliferative response of splenic nylon wool T cells to TBM antigen, was greater in BALB/c and lower in C57BL/6. C3H/He mice showed a significantly high antibody response, but no difference in proliferative response of T cells to TBM antigen, compared with C57BL/6. The histological changes that developed in C3H/He were limited to minimal cell infiltration. Furthermore, nylon wool adherent cells had strong suppressive activity on proliferative response of nylon wool T cells to TBM antigen in low responder C57BL/6. Thus, development of IN was closely related to the cell mediated immunity.

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Western blot technique was applied for qoantifying FDP subfragments. This technique is highly sensitive and can detect 100 pg of D-D dimer and gives high resolution of each subfragment as a stained band on a transblotted menbrane according to the mol, size, without using radioisotpe or affinity purification. We examined plasma or urine from patients with Henoch-Shoenlein Purpura Nephritis (HSPN), Focal Glomeruler Screlosis (FGS) and Orthostatic proteinuria (OP). Results were as follows: Fibrinolytic products including D-D dimer were ditected in HSPN plasma and were increased by Urokinase therapy, however fibrinogenolytic products Y, D and E were not detected or very faint through the therapy. The major component in urinary FDP of both FGS and OP were hsgh moleculer weight (>250KD) fragments, D-D dimer was detected only in the FGS urine suggesting intraglomeruler fibrinolysis.

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Immunoglobulin-E (IgE) production of lymphocytes was examined in 69 children with idiopathic nephrotic syndrome. In cases of an onset or a relapse, 75% of children showed increased IgE synthesis of B lymphocytes without any stimulations. B lymphocytes, treated with cyclohexirnide, produced more IgE in 62% of children with frequent relapses than in healthy controls. The ratio of spontaneous IgE synthesis of B lymphocytes to that of peripheral lymphocytes was elevated in 56% of children with a relapse. B lymphocyte-lines, transformed EB virus, produced high levels of IgE in children with a relapse. These results suggest that B lymphocytes may induce IgE synthesis by depleting T lymphocytes in cases of a relapse, and that they may have many resting IgE precusor cells. In addition, children with frequent relapses were considered to have many activated B lymphocytes for IgE synthesis.

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To clarify the effect of cationization of antigen on platelet activation and glomerular deposition of IC, the relative low dose (50 mg/rabbit) of unmodified (native), moderately cationized and highly cationized ferritin were injected into rabbits as antigens. During acute serum sickness nephritis, the behaviour of platelets was investigated the serotonin release from platelets, platelet counts and platelet-bound immunoglobulin were estimated, and the shape-change were examined by electron microscopy. Platelet counts and intra-platelet serotonin concentration were decreased in all three groups, soon after injection of the antigens. And then second serotonin release from platelets was observed during immune elimination of antigens. In the group given native ferritin, the first serotonin release and decrease of platelet counts were significantly greater than those in highly cationized ferritin. In second serotonin release from platelets (immune release from platelets), plateletbound IgG and significant changes of platelet shape were observed during immune elimination; pseudopod formation and widening of open canalicular system, whereas the serotonin release was not significantly different in any groups. Only in the animals given highly cationized ferritin, deposition' of rabbit IgG and C3 were detected in the glomerulas, Considering that the dose of antigens injected was relative low, IC formed in the highly cationized group might be smaller than those in the native group. It is considered that not only increased vascular permeability, induced by immune release from platelets, but also the characteristics of CIC; size of IC, may also play important roles on the glomerular deposition in acute serum sickness nephritis of rabbit.

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To clarify the effects of membrane materials and anticoagulations on complement activation, the activated components of complements (C3a, C4a, C5a), were examined during hemodialysis (HD) and plasmapheresis (PP). In HD with heparin, cuprophane (C) and saponified cellulose (SC) membranes activated the complement system remarkably via the alternative pathway but only a little activation was noted in HD with polymethylmethacrylate (PMMA) and polyacrylonitrile (PAN). Use of ethylenvinylalcohol (EVA) and cellulose acetate produced moderate complement activation. The classical pathway was suspected to be activated by EVA membrane. Nafamostat mesilate (NM), which inhibited coagulation and complement cascades simultaneously in vitro, prevented HD leucopenia at high doses in animal experiments but failed to suppress complement activation at the clinical dosage with the C and SC membranes. HD with citrate and calcium (Ca) free dialysate partially suppressed complement activation, which indicated a minor contribution of the classical pathway to the complement activation. In PP with heparin, alternative complement pathway was activated by the blood membrane surface interaction in cellulosic membranes, but the activation was lesser than those observed in HD. Classical pathway was suspected to be activated by the membrane surface of polyvinylalcohol (PVA). Other synthetic polymer surfaces brought about little effects on complement system. The high concentration of activated complements in separated plasma (intramembranous complement activation, ICA) was noted in all membrane materials, particularly in PMMA and PVA. Classical pathway was supposed to take the major role in ICA. NM showed no effects on activated complement levels in PP. These results indicate that complement activation during HD and PP is affected by membrane materials and anticoagulations, and in HD the activation can be avoided by proper selection of membrane materials and anticoagulations, In PP, high concentration of activated complement may become a major obstacle especially for donor PP.

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It has been suggested that progression of renal failure can be altered by the manipulation of dietary protein. This study examined the effects of dietary protein on renal function and renal histopathology with or without oral adsorbent AST-120 in chronic renal failure rats induced by partial renal infarction. The uremic rats with average serum creatinine of 1.8 mg/dl were fed with high (36%), normal (24%), or low (14%) protein diet with or without AST-120, in which the content of inorganic phosphate was adjusted to the same level. The survival rates were deteriorated by the increase in dietary protein. Blood urea nitrogen and uremic peak 2a detected by LC were elevated with the increase in dietary protein loading, however, serum creatinine and creatinine clearance were not changed among these three dietary groups. The renal histopathology one month after induction of renal failure showed that high protein dietary groups was significantly lighter in glomerular and interstitial lesions than normal protein groups either with or without AST-120. Judging from the blood biochemistries just before death, high dietary protein group died of uremia with lower serum creatinine and higher creatinine clearance than normal dietary group. Administration of AST-120 for 2 weeks improved the blood level of creatinine, peak 2a and survival rate, but did not improve renal histology. The effect of AST was more prominent in high protein group, but seems to be additive even in protein restricted group, removing some uremic substances through gastrointestinal tract. More detail is to be clarified by further long term study.

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Twenty-two patients (11 males and 11 females) with chronic renal failure (CRF) were treated with rhubarb and traditional Chinese prescriptions. The underlying disease for CRF was 19 cases of chronic glomerulonephritis and 3 cases of polycystic kidney. The patients were placed on a standard diet for 2 weeks, then they were mainly administered decoctions of rhubarb for 4 weeks. After that traditional Chinese prescriptions containing rhubarb were given for another 4 weeks. The initial dosage of rhubarb was 0.5 g/day, and then the amounts were controlled in order to induce one or two soft bowel movements a day. The maximum dosage was 3.0 g/day in cases of rhubarb alone, and 5.0 g/day in cases of traditional Chinese prescrip-tions containing rhubarb. Adverse effects observed with rhubarb administration were abdominal distention, and increase in gurgling sound, abdominal pain, nausea and vomitting. However, these signs became less in accordance with wen-pi-tang administration, i, e., one of the traditional Chinese prescriptions containing rhubarb, ginseng, aconit, licolice and ginger. Following the administration of rhubarb and/or the Chinese prescriptions, the levels of serum methylguanidine, BUN and serum inorganic phosphorus (P) improved significantly. However, the values of serum creatinine (Cr), uric acid (UA), guanidinosuccinic acid, total protein and albumin were not changed remarkably. The amounts of urinary urea nitrogen, Cr and P were significantly reduced following the rhubarb therapy. There was no marked change in the urine volume or in the amount of urinary protein. It also became clear that in the cases whose level of each serum constituent was decreased, the urinary excretion of urea nitrogen, Cr, UA and P was more markedly decreased. The results suggest that the rhubarb therapy possesses the potential to improve azotemia not through an increase of urinary excretion of urea nitrogen, but rather through its anabolic, anti-catabolic effects on nitrogen metabolism. Rhubarb or rhubarb containing Chinese prescriptions will prove to be useful for the treatment of CRF.

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To assess the pathophysiological role of atrial natriuretic polypeptide (ANP) in the regulation of blood pressure, we studied the effect of chronic infusion of synthetic rat-ANP on blood pressure and sodium-water excretion in angiotensin II (A II) -and vasopressin (VP) -infused rats on regular intakes of sodium or on sodium loading with 1% NaCl as a drinking water. When subdepressor dose (150 μg/kg/day) of ANP was infused into the jugular vein by an osmotic minipump (Aizet·) for up to 3 days simultaneously with A II (900 μg/kg/day) infused intraperitoneally by another osmotic minipump, the expected elevation of systolic blood pressure was inhibited little. The antihypertensive effect of this peptide was sustained throughout the experimental period lasting for 3 days in rats on both sodium conditions. It was not accompanied by any changes in body weight, water intake, urine volume and urinary sodium excretion. Additional administration of ANP to rats made hypertensive for 3 days by infusion of A II reduced the blood pressure to a little extent but not to control levels, and the attenuating effect was sustained throughout the experimental period lasting for further 3 days on both sodium conditions. When the same dose of ANP was administered in combination with VP (7.2U/kg/day) for up to 3 days, the expected elevation of systolic blood pressure was completely inhibited. The antihypertensive effect of this peptide was sustained throughout the experimental period in rats on both sodium conditions. It was not accompanied by any changes in body weight, water intake, urine volume and urinary sodium excretions. These results indicate that subdepressor dose of ANP can modulate the hypertension induced by chronic infusion of A II and VP independent of sodium-water metabolism. Therefore it is suggested that ANP may be involved in the regulation of blood pressure via its attenuating effect to A II and VP.

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The effects of α-human atrial natriuretic peptide (α-hANP) on cortisol (F) and aldosterone (Aid) productions were investigated in 6 patients with essential hypertension (4 males, 2 females) during an administration of low dose adrenocorticotropic hormone (ACTH). They received an intravenous infusion of synthetic ACTH (0.7 ng/kg/min) for 60 min on one day and an intravenous infusion of ACTH and synthetic α-hANP (0.1 fig/ kg/min) for 30 min on another day. Synthetic α-hANP infusion significantly reduced blood pressure (p<0.01) and increased heart rate (p<0.01) compared to their basal levels. These changes disappeared within 30 min after the cessation of α-hANP infusion, Although urinary sodium and potassium excretions, and glomeruiar filtration rate were not changed significantly by the infusion of ACTH with α-hANP, urine volume was signi-ficantly (p<0.05) increased after α-hANP infusion. Increment of free water reabsorption significantly (p<0.05) decreased after administration of ACTH with a-hANP compared with that after ACTH administration alone. There was no significant effect on F and Aid productions during the administration of low dose ACTH. Although synthetic α-hANP infusion reduced plasma levels of ADH, there was no significant difference as compared with plasma ADH levels after ACTH administration alone, Although plasma norepinephrine levels were elevated after α-hANP infusion, plasma epinephrine and dopamine levels were not significant changed. Thus it seems likely that the dose of α-hANP used in our study has no effect on F and Aid production during an administration of low dose ACTH in vivo and that the inhibition of free water reabsorption contributes to the diuretic action of α-hANP.

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We studied 375 chronic Hemodialysis patients for evidence of hepatitis B virus (HBV) infection using a first and newly developed second generation monoclonal radioimmunoassays (MRIA). The assay employs high affinity monoclonal Ab produced against antigenic determinants which reside on HBsAg. The assays have a lower limit of detection for HBsAg in serum of approximately 55 and 15 pg/ml respectively. We found that 14 of 375 chronic hemodialysis patients were positive for HBsAg by both polyclonal (PRIA) and MRIA. However, additional 17 were identified as harboring HBV infection only by the MRIA. Thus the MRIA improved the HBV detection rate (PRIA: 3.7 vs MRIA: 8.3%). 10 of the 17 MRIA reactive patients had HBcAb as evidence of HBV exposure. 24 out of 145 HBcAb positive patients were found to have HBsAg by MRIA. We conclude that there are hemodialysis patients with HBV infection not detected by conventional polyclonal radioimmunoassays.

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The effect of mercury administration and simultaneous administration of mercury and selenium on intrarenal concentration of these metals and activity of glutathione peroxidase (GPx) of rat kidney were studied. Control rat was fed by laboratory chow freely. Mercury was administered by mercury vapor at a concentration of 8.0 mg/m³ for a hundred minutes/day during 6 weeks. Selenium was given by 10 ppm sodium selenite tap water, Rat was killed by decapitation and kidney was removed to homogenize by Potter-Elvehjem grinder at 1, 000 rpm for 1 minute. The homogenate was centrifuged to separate mitochondrial and lysosomal fraction at 25, 000×g for 20 minutes, then microsomal fraction was separated from supernatant at 100, 000×g for 60 minutes. GPx was measured by the method of Nakamura et al, Mercury content was assayed by the reducing vapourization method, selenium by the fluorometric method with 2, 3-diaminonaphthalene and protein content by the method of Lowry et al. In mercury vapor inhalted rat, intrarenal concentrations of mercury and selenium were remarkably increased, Further increase was observed in the rat simultaneously administered with mercury and selenium. It was found that mercury mainly concentrated in mitochondrial and lysosomal fraction in all experiments, On the other hand, selenium was mostly found in the supernatant of mercury vapor inhalated rat. But, in the rat simultaneously administered with mercury and selenium, selenium was remakably increased mitochondrial and lysosomal fraction. GPx activity of mercury vapor inhalated rat was significantly depressed in microsomal and supernatant fractions comparing to those of control rat. On the other hand, no significant difference of GPx activity was observed between control rat and the rat simultaneously administered with mercury and selenium. By these results, it was suggested that selenium was effective in prevention of decrease in GPx activity of supernatant and microsomal fractions of mercury vapor inhalated rat.

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Lipid metabolism of liver and kidney were examined in daunomycin induced nephrotic rats.1) Serum cholesterol were increased from 2 to 5 weeks in daunomycin induced nephrotic rats.2) The daunomycin induced nephrotic rats showed the increase of Acyl-CoA: cholesterol acyltransferase (ACAT) activity and cholesterol esters content in liver icrosomes.3) Cytosol cholesterol esterase in liver and kidney were decreased in daunomycin induced nephrotic rats.4) Lysosome cholesterol esterase activity in liver and kidney were higher in daunmycin induced nephrotic rats than control rats. These results show that the increased cholesterol esters in liver and kidney are caused by decreased cytosol cholesterol esterase activity in daunomycin induced nephrotic rats, and the accumulation of cholesterol esters in liver seems to result from the increased ACAT activity. Increased lysosome cholesterol esterase activity in liver and kidney of daunomycin induced nephrotic rats suggest that incorporation into tissues from lipoprotein-cholesterol esters may be increased.

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The relations between autonomic function and hemodialysis-induced hypotension were investigated in 85 patients undergoing maintrnance hemodialysis treatment using the changes in coefficient of variation of R-R intervals (CVRR) and using the changes in plasma colloid osmotic pressure (COP) during hemodialysis. To evaluate the effect of Medical Antishock Trouser (MAST) on the hemodialysis-induced hypotension, we observed 22 patients undergoing maintenance hemodialysis treatment. As a result, CVRR decreased with age in hemodialysis patients as well as in healthy subjects. A highly significant positive corelation was observed between the reduction of CVRR and the increase of incidence of hypotension during hemodialysis. Intermittent MAST inflation inhibited early hypotensive episodes (within the first 2.5 hr) but not inhibited late hypotensive episodes. It is suggested that autonomic insufficiency may contribute to the pathogenesis of hemodialysis-induced hypotension, and MAST are effective in managing the early hypotensive episodes.

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