As a model of human membranous nephropathy, passive Heymann nephritis (PHN) was established in female Sprague-Dawley rats by intraperitoneal injection of rabbit antirat FxlA serum. Morphological studies with lightmicroscope (LM), immunofluorescent microscope (IF) and electron. microscope (EM) were done 1 hour through 6 months after anti-Fx1A serum injection. Also the presence of a FxlA like antigen in rat glomeruli was studied using indirect immunofluorescence and solid phase radioimmunobinding test. Two phases were noticed. First (heterologous) phase was characterized by mild proteinuria on day 5 through 10 (54±9mg/day, mean+S.D.) and diffuse granular staining of rabbit gamma-globulins and rat C3 along glomerular capillary walls. Second (autologous) phase was accompanied with more prominent proteinuria (152±28mg/day), the deposition of rat IgG in addition to rabbit gamma-globulins and the disappeappearance of rat C3. Observation on EM disclosed the sequential changes of the subepithelial deposits whichh were similar to the staging of human membranous nephropathy. Initially amorphous deposits, then well demarcated ones associated with thickening of GBM and finally lucentt ones were observed. By indirect IF on the frozen sections, normal rat glomeruli did not show any specificc binding with anti-FxIA antibody even after removal of surface glycoproteins by neuroaminidase. However, isolated rat glomeruli and their sonicated fractions binded the antibody significantly by IF and solid phase radioimmunobinding test. Thus, it is concluded that the initial mechanism of PHN is in situ immune complex formation between anti-FxIA antibody and a glomerular FxIA like antigen, while superimposed deposition of autologous antibodies against rabbit gamma-globulins seems to play an additional important role in persistance of PHN.

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The components of the immune complexes deposited in the lesion of lupus like glomerulonephritis in NZB/W F₁ mice been recognized to be mainly composed of DNA as an antigen and anti-DNA antibodies. However, there are many problems to be elucidated the components of immune complexes. Recently we developed new procedure to dissociate immune complexes in glomeruli by applying various reagents on the frozen kidney sections. Present paper described is to study a composition of immune complexes in glomerulii of NZB/W F₁ mouse and also an activity of antinuclear antibody in the serum and eluate from the mouse kidney cross-reactive with mouse or rat IgG, mainly Fc fragments. Granular staining for mouse IgG in the mesangium and glomerular capillaries were observed almost of all 6 month- to 12 month-old NZB/W F₁ mice. These frozen sections of the kidney were performed the dissociation test of mouse IgG, IgM and C₃ from the glomeruli. The frozen sections were incubated with 5% of mouse IgG, mouse Fc fragment (frg), rat IgG, rat Fc frg, human Cohn fraction 11 (Frll), 0.5% of DNA and nucleohistone and PBS for 6 hours at room temperature chang in PBS, and then these specimens were stained with FITC-conjugated antimouse IgG, IgA, IgM and C₃ antibodies. Among these, the IgG deposits in the glomeruli disappeared or decreased following incubation with mouse IgG and rat IgG, especially with mouse Fc frg and rat Fc frg. On the contrary, the IgG deposits did not disappeared following incubation with human FrII, rabbit FrII, DNA, nucleohlitone and PBS. The activity of antinuclear antibody (ANA) was studied whether ANA was crossreactive with other reagents, mainly mouse IgG. Activity of ANA in the serum disappeared after incubation with mouse IgG, mouse Fc frg, rat IgG and rat Fc frg, but not with human IgG and rabbit IgG. Activity of ANA in the eluate disappeared after incubation with mouse IgG, mouse Fc frg, rat Fc frg, DNA and nucleohistone, but not with human and rabbit IgG. These results suggested that the immune complex deposits in the glomeruli consisted of IgG (presumably denatured)-IgG ANA/IgG rheumatoid factor.

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Investigation of antigens in immune-complexes deposited in glomeruli has been performed on elution study of the antibody or on application of FITC-labelled antibody corresponding to the antigen. In the present study described, detection of the antigen deposited in the glomeruli, using three kinds of experimental glomerulonphritis (GN) (PHN, CSS and MN) in rats were established by following two procedures. The first is an investigation of antibody activity in serum from the animal immunized with diseased glomeruli. The sera obtained from the rabbits immunized with the extract from glomeruli with PHN were performed indirect immunofluorescence on the kidney section of rats. The rabbit lgG was found to react with brushborder of the proximal tubules. The serum obtained from rabbit immunized with glomeruli of CSS was tested by Ouchterlony test against BSA. Single precipitation line was found to form between the immune serum and BSA.. The serum obtained from rat immunized with GBM of Masugi nephritis was also tested against rabbit IgG. Single precipitation line was found to form between the serum and rabbit IgG. The second is dissociation of IgG deposits or the associated antigen from glomeruli by applying excess of antigen on the frozen sections of these diseases. Although the BSA staining in glomeruli of CSS disappeared following incubation of excess amount of BSA, the IgG remained in the glomeruli, suggesting that binding site of IgG was not only on the BSA, but also on a different site in the glomeruli . The staining of rat IgG in the glomeruli from second phase of Masugi nephritis disappeared following incubation with rabbit IgG. In passive Heymann nephritis, rabbit IgG was not even abolished following inc ubation with Fx1A, suggesting that rabbit IgG may bind tightly with the antigen on the epithelial cells. These results indicate that the immunization procedure of the diseased glomeruli or the incubation of excess of antigen will become very usefull to detect an antigen in the glomeruli.

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Metabolic state and pituitary-thyroid function were investigated in patients with endstage renal failure in order to clarify the pathomechanism of the thyroid disturbances. Patients, who were all free from thyroid diseases, were divided into three groups; the non-dialyzed, azotemic patients with high serum creatinine level (1.5-8mg/dl) (group I), the patients prior to dialysis therapy with uremic symptoms (group II), and the patients receiving hemodaalysis for more than six months (group III). 211 patients in group III showed weakness (75.4%), cold intolerance (63.6%), memory impairment (60.6%), constipation (55%), dry skin (34%), and others. These symptoms are very similar to hypothyroidism. Patients in group I demonstrated lower normal value of total thyroxine (TT₄), low value of total triiodothyronine (TT₃) (P<0.001), normal values of free T₄(FT₄) and reverse T₃ (rT₃), and slightly high basal value of thyrotropin (TSH) (P<0.025). Patients in group II revealed much lower value of TT₄, TT₃, FT₄(P<0.001) and high basal level of TSH (P<0.005). Patients in group III disclosed low values of TT₄, TT₃, FT₃ (P<0.001), rT₃ (P<0.005) and normal level of TSH. Moreover, there were significant reductions in TT₃/TT₄ ratio levels in all groups compaired with normal controls. The pituitary secretion of TSH after thyrotropin-releasing hormone (TRH) injection was very poor in groups I and II. Although group III patients showed the moderate recovery of TSH secretion reserve, the maximum increment of serum TSH was insufficient in comparison with normal controls. Hemodialysis patients showed significantly low level of the thyroidal ¹³¹I-uptake after oral and intravenous radioiodide administration. Basal metabolic rate was -19.4±8.4% and the contraction phase of the Achilles tendon reflex was significantly elongated in these patients. These data indicate that patients with chronic renal failure have the abnormalities of the pituitary-thyroid axis and appear to be in hypometabolic state.

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Humoral immunity in chronic renal failure (CRF) was studied using a T cell-independent B cell mitogen Staphylococcus aureus Cowan I (STA) or a T cell-dependent B cell mitogen pokeweed mitogen (PWM). Results are as follows. 1) The proliferative responsivess or in vitro immunoglobulin (Ig) productive capacity of peripheral blood mononuclear cells (PBMC) stimulated with PWM or STA was markedly decreased in patients with CRF as compared with that in healthy controls. 2) The decreased proliferative responsiveness of PBMC to STA from the patients with CRF was markedly restored in some patients by depletion of phagocytic cells, while in others no restoration was observed indicating the possibility of functional B cell defects. 3) Furthermore, the decreased response to various mitogens including STA was not restored with some exceptions by the addition of indomethacin which inhibits prostaglandin E sythetase. The results described above indicate the impaired humoral immunity in patients with CRF, and may lead to frequent episodes of infections or other immunological complications.

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Experimental ischemic acute renal failure model of rabbit was prepared by the excision of right kidney, and the clamping of left renal artery and vein for one hour. Serum concentrations of UN, creatinine, electrolytes and organic acids in the uremic rabbits under intravenous infusion of glucose alone, or glucose with essential amino acids were determined 2 days after clamping renal vessels. In uremic rabbits with infusion of essential amino acids, BUN to creatinine ratio and serum potassium concentration were decreased as compared with the uremic rabbits with glucose infusione. In uremic serum of rabbits with infusion of essential amino acids, many benzoic compounds such as phenylacetylglycine, 3, 5-dihydroxybenzoic acid, 3, 4-dihydroxyphenylacetic acid, 4-hydroxy-3-methoxymandelic acid were detected using gas chromatography-mass spectrometry in higher concentrations as compared with uremic rabbit serum with glucose infusion alone. These benzoic compounds seem to be derived from the metabolites of phenylalanine in the essential amino acid solution.

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A 32 year old female patient on maintenance hemodialysis revealed hypocalcemia (s-Ca; 5.6mg/dl, ionized Ca; 3.4mg/dl) during 5 years in spite of the normal levels of alkaline phosphatase. No increase in serum iPTH values was found (C-terminal assay; 0.82ng/ml, N-terminal assay; 0.1ng/ml) and serum levels of 25 OHD₃ and 1.25(OH)₂D₃ were 15.0ng/ml and 11.0pg/ml, respectively. Radiological bone survey revealed no pathological changes including secondary hyperparathyroidism and/or osteomalacia. Bone biopsy showed osteoclastic bone resorption within some parts of the trabeculae and no marrow fibrosis. Hypocalcemia was seemed to be caused by persistent hyperphosphatemia and decrease in serum 1, 25 (OH)₂D₃ levels. However, it could not support so-called Bricker's trade-off hypothesis that overt secondary hyperparathyroidism was not observed in spite of the nersistent hvperphosphatemia before the start of hemodialysis treatment.Coexistence of hypocalcemia, inappropriately low levels of serum iPTH, and lack in radiologically overt secondary hyperparathyroidism made us speculate the presence of some unknown humoral factors, which suppress PTH secretion, or disturbed responsiveness of PTG to hypocalcemic stimuli. Further research on PTH synthesis and secretion mechanism will make it easy to understand the pathogenesis of our case more precisely.

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After 25mg (or 50mg) of captopril was administerd orally, blood pressure, plasma renin activity (PRA), plasma aldosterone concentration (PAC) and plasma captopril concentration were measured in nine patients with renovascular hypertension. Samples for the measurement of PRA were obtained from each of the two renal veins and from the femoral vein before, 30 minustes and 60 minutes arter captopril. Captopril induced a significant decrease in blood pressure. PRA was significantly increased and PAC was slightly decreased. The renal vein PRA ratio (diseased/contralateral, RVRR) increased 1.77±0.25 to 4.79±1.49 at 30 minutes and to 3.59±0.75 at 60 minutes after captoril. PRA after captopril showed a significant correlation with PRA before captoril. In patients with bilateral renal artery stenosis, the increase of RVRR was small as compared with those with unilateral lesion. There were no relationships between the change in RVRR and the changes in blood pressure, PRA or Plasma captopril concentration. The contralateral renal vein PRA/peripheral PRA (Rc/p) was decreased from 1.12±0.05 to 1.00+0.15 at 30 minutes and to 0.91±0.10 at 60 minutes after captopril. Thus, the contralateral kidney in this disease appeared not only to suppress the release of renin but also to excrete renin, especially after captopril administration. Surgery was performed in five patients. Hypertension was cured or improved in four patients. However, surgery failed to lower blood pressure in a patient with unilateral renal artery lesion, although RVRR was increased after captopril. In this case Rc/p was markedly increased. It is concluded that the change in RVRR after captopril would give a support to the diagnosis of renovascular hypertension and the change in Rc/p after captopril would predict the result of surgery in these patients. This is a safe and accurate procedure for the detection of renin mediated renovascular hypertension.

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An 11 year-old girl suffered from diffuse uveitis with tubulointerstitial nephritis (TIN) was reported. 67Ga scintiscan revealed that the uptake increased in lacrimal glands, parotid glands and kidneys. The sialography of the left parotid gland revealed pseudosialoectasia. Minor salivary gland biopsy was performed and infiltration of plasma cells in the interstitium with degeneration of its ductal epithelium was noticed. Renal biopsy showed that the renal interstitium was markedly infiltrated with plasma cells and immunoglobulins (mainly Ig G) were were found in the plasma cell cytoplasma by IF method. The tubular basement membrane was slightly thickened, Ig G and electron dense deposits were found to be exist in the tubular basement membrane by IF method and electron microscope. Although the pathogenesis of this case is not well-known, it may be an immune-complex disease resulted from B cell hyperreactivity, just like Sjögren's syndrome, since the pathological findings of this case was similar to that of Sjogren's syndrome. In the past, 9 cases of TIN associated with uveitis were repoted and some immunological abnormalities were demonstrated in each case. However, to determine a diserse entity for these cases seems to be difficult, because their pathological findings were not all the same.

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Blood levels of α₂-plasmin inhibitor (α₂-PI) in children with various renal diseases were measured, and changes of those during combined use of heparin and urokinase were investigated in 9 children with several types of nephritis. The following results were obtained. (1) The blood levels of α₂-PI in nephrotic stage of pure nephrosis were lower than those in other nephritis, which were followed by a sharp rise with simultaneous disappearance of proteinuria and then returned to normal levels or more. The blood levels of α₂-PI in nephrotic stage of CGN remained generally normal as well as those in nonnephrotic stage of it. On the one hand, the blood levels of α₂-PI in the early stage of 2 cases with post-streptococcal AGN were apparently lower with gradual increase of levels during the course of the disease. (2) Nine cases with nephritis resistant to the conventional treatment were treated with combineduse of heparin and urokinase (UK). By the administration of UK, the blood levels of α₂-PI fell down to less than 60% of the normal in 4 cases, 2 cases of whom improved clinically. Although, other inhibitors of plasmin such as α₂-macroglobulin and α₁-antitrypsin were not fluctuated significantly during the treatment. Accordingly, it might be assumed that the blood levels of α₂-PI during this treatment reflected accurately fibrinolytic state in blood. Moreover it was apparent that the dosage and speed of administration of UK influenced the activation of fibrinolytic state.

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In an attempt to clarify the alteration in nitrogen metabolism of the patients with chronic renal failure. plasma amino acid and keto acid concentrations were measured. Fasting plasma free amino acid concentrations were measured in 13 patients withh chronic renal failure (CRF), 19 with nephrotic syndrome, 5 with renal transplantation and 21 controls. In CRF, plasma amino acid concentrations, especially 1-methylhistidine, arginine, taurine, hydroxyproline, glutamine, citrulline, and alanine were elevated, whereas essential amino acids, such as valine, threonine and tryptophan were markedly decreased. In primary nephrotic syndrome, no significant difference in plasma amino acid concentrations were found, whereas lower plasma amino acid concentrations were observed in nephrotic syndrome secondary to diabetes with more advanced renal dysfunction. Plasma amino acids determination under the two different energy intake with low protein diet in 10-14 days (Diet I: 25kcal/kg energy and 0.6g/kg protein, Diet II: 40kcal/kg energy and 0.6g/kg protein), were also carried out in patients with CRF. Under the Diet II, serum urea nitrogen declined and plasma amino acids increased in comparison with. Diet I. In addition, plasma concentrations of 3-branched chain ..-keto acids (BCKA) were determined by means of high liquid chromatography in 8 patients with CRF, 7 nephrotic syndrome and 5 control subjects. Significant lowered BCKA concentrations observed in patients with CRF, as compared to controls under the same diets. Conversion of α-keto valine to valine following the oral administration of the analogue (0.1g/kg) were also measured in CRF and controls under the Diet I and II. As a result, increase in plasma keto valine concentration was observed after the oral administration, followed by increase in plasma valine. The conversion rate appeared to be accelerated under the Diet II. Therefore, it is indicated that reutilization of urea nitrogen with keto analogue may be promoted under the adequate calorie and low protein diets.

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Protein binding inhibitor (abbreviated PB-Ix), which may be presumed to accumulate because of poor excretion from kidney, competes with certain kinds of chemical substances, binds to binding sites on the surface of albumin, and inhibits the detoxication, transport and reservoir function of plasma protein. Thus we supposed it has some effect on cell function and may be one of the immunosuppresive factors. Toxicity of the PB-Ix to living cells was examined, while blastformation of normal lymphocytes mixed with the PB-Ix was checked by ³H-Thymidine uptake. Infuence of PB-Ix on the phytohemagglutinin (PHA)-receptor of lymphocyte was studied by using fluorescenced PHA. As the concentration of the PB-Ix increased from 5mg/ml to 20mg/ml, inhibitory effect on 3H-Thymidine uptake of cultivated lymphocytes became apparent. The lymphocytes pretreated with PB-Ix apparently showed the suppressive effect on blastformation, but the lymphocytes stimulated preliminary by PHA did not revealed suppressive effect although PB-Ix was added afterwards. And also lymphocytes pretreated with PB-Ix had markedly lower activity of fluorescence of PHA than control. These results suggested to us that PB-Ix binds to the PHA-receptors of lymphocytes on their surface and therby inhibits their mitogenic ability.

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Fifty-one diabetic patients who underwent 66 radiographic procedures using contrast media were studied with regard to the risk factors of contrast media induced acute renal failure retrospectively. Thirteen episodes of acute renal failure in 12 patients were compared to 53 uneventful radiographic studies. Prior renal insufficiency, hypertension and nephrotic syndrome were thought to be important risk factors clinically. Likewise, raised serum creatinine (over 2mg/100ml), hypoalbuminemia (under 3g/100ml) and nephrotic range proteinuria (over 3.5g/day) were important risk factors. We did not find any significance for risk factor as to age, duration of diabetes, insulin requirement, fasting blood sugar, BUN, serum uric acid and dosage of contrast media. Also, concomitant use of diuretics or antibiotics was not more frequent among affected patients, We could not clear the importance of hydration because of difficulty of getting information retrospectively. Of the 13 episodes of contrast media induced acute renal failure, 6 episodes (46%) ended in irreversible renal failure and 3 of them required maintenance hemodialysis within 6 months. However, there is no significant difference in above mentioned parameters between reversible and irreversible group.

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