The purpose of this study was to resolve how the alteration of polyanion layer in GBM influences upon the onset and progression in immune complex glomerulonephritis. The effect of protamine sulfate (P) and heparin (H) on apoferritin (AF) induced glomerulonephritis was studied histologically. In this experiment, 37 male Swiss albino mice were classified into five groups, that is, I : P only, 5 cases, II : AF only, 9 cases, III : P+AF, 9 cases, IV: H+AF, 9 cases, V : no administration, 5 cases. They were administrated 4 mg of AF and 50U of H, one time and 1 mg of P, two times daily by intraperitoneally for 3 weeks. As a result, focal endocapillary proliferative glomerulonephritis was recognized in group of II, III and IV. Each group showed prominently demonstration of the vulnera-bility of glomerular lesion in the juxtamedullary cortex. Group II was a considered as a highest degree of glomerular alteration and showed significant differences of histological findings as compared with another groups. Proteinuria and IF findings also had a similar tendency in group II. As regards alteration of polyanion layer in GBM, loss of polyanion was recognized at the glomeruli with severe cellular proliferation in each group. And group III was relatively severe on another groups, but as the result of statistical analysis, there was not significant differences among each group. By a comparison of group II and group IV, it was shown that group IV produced proteinuria and glomerular lesion but it was a lesser degree rather than group II. Moreover, accumulation of fibrin in the glomerular capillaries was severely present but there were little differences between group II and group IV. These findings are suggested that heparin kept anionic charge may be effective in part for diminishing in quantity of protein excretion and also heparin which has the other effect of anticoagulative reaction, such as antiproliferative effect, influences upon the cellular proliferation in the glomeruli.

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To examine the alterations of glomerular basement membrane (GBM) and the character of electron dense deposits (EDD) in various glomerular diseases, electron microscopic findings using periodic acid methenamine silver stain (PAM stain) and immunohistochemical technique were analyzed. Membranoproliferative glomerulonephritis (MPGN) type I(4 cases), type II (2 cases), type III (1 case), membranous nephropathy (3 cases), lupus nephritis (1 cases), IgA nephropathy (2 cases) and Henoch-Schoenlein's purpura nephritis (2 cases) were studied. PAM stain was an useful probe for electron microscopic analysis, by which were obtained the more precise findings of GBM changes at the severely proliferative segments and the portions with numerous EDD. Especially, this stain was essential for histological diagnosis of MPGN type III. It was suggested that ribbon-shaped EDD of MPGN type II might be neither immunologic deposits nor degeneration products of GBM itself, but glycoprotein-rich subjects from systemic diseases such as metabolic one.

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We attempted to clarify main factors of edema formation in nephrotic patients. Simultaneous measurements of plasma volume (PV), plasma renin activity (PRA), plasma aldosterone (PA), α-human natriuresic peptide (α-hANP) and urinary sodium excretion (U_NaV) were made at edematous and edema-free states in seven patients with minimal change (MC), seven with membranous glomerulonephritis (M) and eight healthy subjects (C). In addition, an oral water load of 20 ml/Kg body weight was performed in 6 of MC, 6 of M and 3 C. Urine volume/water intake (UV), free water clearance (C_water), sodium clearance (C_Na) and plasma antidiuretic hormone (ADH) were determined before and 2, 4 and 6 hours after water load. PV on edematous state increased significantly in nephrotic patients as compared with C, and decreased to normal level at edema-free state. U_NaV in MC on edematous state was significantly lower than M and C, and showed negative correlation to PV. And, U_NaV increased in MC during edema removal. Althogh PRA on edematou state was markedly higher and decreased on edema-free state in MC, PA remained within upper range of normal during edema removal. In MC α-hANP also remained within normal range, but in M it was significantly higher on edematous state and decreased durinc edema removal. UV_s during first 2h in C, MC and M was 60.5%, 27.8% and 39.5%, respectively. There was not a significant difference in C_water before water load among MC, M and C. In C, C_water increased in the first 2h after water load, however, in MC and M, lower level of C_water persisted. C_Na in C increased in response to diuresis, but C_Na of nephrotic patients slightly decreased and did not change during water load. The proximal tubular sodium reabsorption in the 4h after water load in MC and M was higher than that of C, and did show negative correlation to UV and C_Na. ADH did not change after water load in all of the subjects. It was concluded that nephrotic patients have the decreased excretion of water and sodium. The impairments of diuresis and natriuresis may be attributed to the increase of proximal tubular sodium reabsorption. There was a difference in impairments of diuresis and natriuresis between MC and M, which would well explain the mechanism of that edema was more severe in MC than M.

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We investigate the long-term outcome of 15 nephrotic patients with primary mem-branous nephropathy. The effect of treatment with corticosteroid initially with immunosup-pressants (100 mg/day of either cyclophosphamide or azathioprine) were observed for up to 101 months (mean 56 months). Their mean age at start was 41 years old. At the 6 month' period, there was 2 patients in complete remission and 4 in incomplete remission type I. However, there were additional 4 in complete remission at 2 years' period. The number of patient in complete remission increased to 10 with 4 in incomplete remission type I most recently. And there was only one patient still in persistent nephrotic state and none had worsening renal function at final observation. No major side effect was encountered in any patients. The effect of treatment correlated neither with morphological stages nor with the clinical findings. We conclude that the long-term use of corticosteroid initially with immunosuppressavt is effective in most nephrotic patients with primary membranous nephropathy. Additionally we noted focal segmental glomerulosclerosis (FGS lesions) in 4 patients with membranous nephropathy. They were aged and showed clinical characteristics of hypertension. Patients with FGS lesions on membranous nephropathy should be further analyzed.

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Previous studies show that prostaglandins (PGs) are present in the glomerulus and have the effects on glomerular hemodynamics. I studied on PG biosynthesis in both glomeruli obtained from rats with streptozotocin-induced diabetes mellitus (DM) and cultured rat mesangial cells. Glomeruli were isolated 2 weeks and 12 weeks after strep-tozotocin administration. Production of PGs was determined by gas chromatography masspectrometry after incubation of glomeruli under basal condition or in the presence of arachidonic acid (C20: 4). In the early stages of DM, the production of PGE₂, PGF₂ α, 6-oxo-PGF₁ α and PGD₂ in the presence of C20: 4 was significantly greater than in the control. In basal condition, PGE₂, PGF₂ α, 6-oxo-PGF₁ α and PGD₂ were also increased in the diabetic rats. The rate of PG production under C20: 4 condition was related directly with the degree of hyperglycemia. GFR was increased in these diabetic rats. In the chronic stages of DM, the production of PGE₂, PGF₂ α, 6-oxo-PGF₁ α, PGD₂ and TXB₂ in the presence of C20: 4 was significantly greater than in the control. However, in basal condition, the production of PGE₂ and PGD₂ was decreased comparing with the control. GFR was not significantly defferent between the control and DM rats. High osmolality condition by glucose or mannitol stimulated all PG synthesis of cultured mesangial cells. These findings suggest that PG metabolism would play an important role in the progression of diabetic glomerular disturbances.

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It has been well known that urinary excretions of total N-acetyl- β-D-glucosaminidase (NAG) as one of tubular proteinuria are usually increased in the patients with nephrotic syndrome. But nephrotic patients are merely associated with tubular dysfunctions, so it is unclear the course of increased urinary NAG excretion at the active stage of this disease. The purpose of this paper is to define the origin of the urinary NAG in nephrotic syndrome by using the rats models due to puromycin aminonucleoside (PAN) administration. Nephrotic state were induced on the fifth day after only PAN 15 mg/100 g B. W subcutaneously injected to the rats in isolated metabolic cages. Serious analysis of NAG isozyme was performed by operating the fast protein liquid chromatography (FPLC, Pharmacia) connected with DEAE cellulose column. Then NAG activity was determined by m-cresolsulfophthalein-NAG (MCP-NAG) method in urine, glomerular and tubular tissue isolated by pore sieving. Urine was dialyzed to exclude low molecular weight substances and renal tissues were homogenized and centrifuged to get the supernatants for estimating NAG isozyme. As the result three components of NAG which were B, I+II and A isozyme were detected in urine and elutes from renal tissue. NAG B isozyme with nontreated rats were 9.2±1.5%, 46.3+7.4% and 17.5+2.6% in urine, glomeruli and tubular tissue respectively. The B/I+II+A isozyme ratio in urine were changed from 0.10±0.01 to 1.06±0.21 on the active stage of PAN nephrotic rats. According to the high level of B/I+II+A in glomeruli on the early stage of nephrotic rats, it might be supposed that NAG were discharged from the glomeruli. In conclusion, it is clearly defined that the source of NAG is partly derived from glomeruli in patients with nephrotic syndrome.

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Numerical and functional assay of suppressor cells was studied in 20 chronic hemo-dialysis (HD) patients. Most of the patients were clinically stable, though treated with HD for less than one year and had no history of blood transfusion. Peripheral blood mononuclear cells were classified into total, inducer/helper and cyto-toxic/suppressor T cells by the monoclonal antibodies, OKT3, OKT4 and OKT8, respectively. Suopressor cell activity was measured by means of Concanavalin A induction (ConA-SCA). The percentage of OKT3 and OKT4 positive cells were 54.5±11.1% and 33.4±8.8% respectively in the patients, significantly lower than that of controls (60.2±6.9% and 37.9±8.8%). However, no significant difference in the percentage of OKT8 and OKT4/ OKT8 ratio was noticed between the patients and controls. ConA-SCA was significantly reduced in the patients (16.0±19.4%), compared to the controls (32.3±23.5%). The discrepancy between the surface markers and function of T cell subset suggests that OKT4 cells may be heterogenous and possibly have the suppressor function. Considering decreased suppressor cell activity in non-transfused patients, blood transfusion may enhance suppressor cell activity.

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Natural killer (NK) activity was studied in 24 chronic hemodialysis (HD) patients by the method of ⁵¹Cr releasing assay using K-562 cell. There was no significant difference in NK activity between the patients and controls. No significant difference was also noticed in the augment effect of exogenous interleukin 2 to NK activity between the patients and controls. HD duration for more than 60 months had some effect on NK activity, although it was not related with age, transfusion, serum urea nitrogen, creatinine or β₂ microglobulin, vitamin D₃ and the cause of renal failure. This might be related with higher incidence of malignancy in those patients.

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The production of and reaction to interleukin 2 (IL-2) were studied in 55 chronic hemodialysis (HD) patients. Production of interleukin 2 stimulated with PHA in peripheral blood mononuclear cells in HD patients (0.67±0.75 unit/ml, Mean± SD) was almost the same as that of healthy controls (0.43±0.37 unit/ml). Response to exogenous human IL-2 was significantly reduced in HD patients (32, 859±6, 807 Δcpm) compared to that of healthy controls (38, 857±10, 341 Δcpm, p<0.05). The production of as well as response to IL-2 did not show any relationship with age, HD duration, serum urea nitrogen, creatinine, β₂ microglobulin, vitamin D₃, transfusion and the causes of renal failure. These results suggest that the abnormality of IL-2 receptor may be one of the causes of low immune respnse in HD patients.

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To clarify the influence of sodium intake on antihypertensive effect of calcium antagonist, changes of blood pressure and humoral factors after a single oral administration of 40 mg nicardipine were evaluated in 15 subjects with essential hypertension under high-, normal-, and low-sodium regimens (mean 24-hour urinary sodium excretion: 320±24, 147±7, 27±6 mEq, respectively). Nicardipine induced a significant reduction of mean blood pressure and increase in heart rate. The change of mean blood pressure after nicardipine was negatively related to pretreatment mean blood pressure under three sodium intakes (p<0.01). The slopes of the correlation lines for high-, normal-, and low-sodium regimens were -0.61, -0.69, and -0.52, respectively, without a statistical significance. Nicardipine brought about significant increases in plasma renin activity and plasma nore-pinephrine, but no changes in plasma epinephrine or serum aldosterone concentration. These results suggest that the magnitude of the untreated blood pressure and thereby the peripheral resistance seems to be major determinants of the blood pressure fall by calcium antagonist, and failure to increases of aldosterone and epinephrine in the face of peripheral vasodilatation may have shared in part in the the antihypertensive effect of this drug.

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We measured serum laminin in the patients with renal diseases and compared them with normal levels (n=71, 1.35±0.19). Serum level of laminin in the patients with minimal change nephrosis at the remission phase (n=6, 1.41±0.15) and IgA nephropathy (n=25, 1.27±0.18) showed no significant differences compared with normals. However, minimal change nephrosis at the nephrosic phase (n=4, 1.82±0.14), membranous nephro-pathy (n=11, 1.70±0.37), diabetic nephropathy (n=16, 1.76±0.49), lupus nephritis (n=9, 1.88±0.64) and renal carcinoma (n=30, 1.67±0.51) had significantly high (p<0.01) levels of serum laminin concentrations. There are no relationship between serum laminin level and creatinine clearance. This result suggests that serum laminin level may indicate the alteration of the glomerular basement membrane metabolism in these renal diseases.

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Electrothermal atomic absorption spectrophotometry has been used to analyze aluminum in various kinds of materials. However, several technical problems has to be solved to apply this method on determining aluminum in biological materials, including instrument setting, standardization procedure and eliminating contamination. This method consists of drying, ashing and atomizing material, all of which are performed by graphite furnace. Reproducibility of measurement largely depends on the steps of drying and atomizing. Interferences could be elininated to a great degree in ashing step when the temperature was above 1100°C and ashing time was 20 seconds. In order to eliminate or to reduce background of measurement and chemical interferences, matrix modifiers have been used in atomic absorption spectrophotometry. We employed serum based standard method for serum sample to avoid using multiple kinds of modifirs which causes high reagent blank. In this method, reagent blank has not been detected. Precision of this method whithin-and between-assay were CV=2.7% and 4.2%, respectively. Mean recovery of externally added aluminum was 101.8%. We also investigated the possible sources of aluminum comtamination. In this study, we succeeded to establish highly accurate and reliable method for measuring serum aluminum concentration. This technology enables us to make an accurate diagnosis on aluminum toxicity in the patients with chronic renal failure.

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Serious complications such as pulmonary edema may occur due to a marked decrease in colloid osmotic pressure (COP) secondary to hypoabuminemia in plasmapheresis. It is important to adequate albumin concentration of the substitution solution to avoid hypoal-buminemia in double filtration plasmapheresis (DFPP). In this study, DFPP was per-formed by using substitution solution with either the high concentration (12.5% albumin; group H) and relatively low concentration (5% albumin; group L) for 44 times in 12 patients. COP, concentrtion of total serum protein (protein) and serum albumin (albumin) before, after and at an interval of 15 minutes during DFPP were measured, and compared with the values before DFPP as control. In group L, COP, protein and albumin were significantly lower compared with those before DFPP. In contrast, in group H, COP was significantly lower than control for 45-90 minutes during DFPP, protein was significantly lower 75 minutes and after, whereas albumin significantly increased during and after DFPP. Protein, albumin and COP in group H were also significantly higher than those in group L during and after DFPP, in spite of no significant difference in control values between group H and L. It is concluded from our results that DFPP can be performed more safely by monitor-ing serum COP values and with less change in serum albumin level using substituion solution with relatively high albumin concentration.

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We investigated the transport systems of phosphate(Pi), D-glucose, L-alanine, and sodium ion in renal brush border membrane vesicles (BBMV) prepared from 5- or 12-wk-old SHR and Wistar-Kyoto rats (WKY). Rat renal cortex BBMV were isorated by a Mg²⁺ precipitation and purified by controlled pore glass column chromatography. In the initial uphill phase at 20 sec., the uptake rate of Pi, D-glucose, and L-alanine by BBMV prepared from SHR were lower than that prepared from WKY in the presence of sodium ion gradient. The Pi concentration of incubation medium was varied from 12.5 to 300 μM, and apparent Km and V value were calculated from regression lines obtained from Eadie-Hofstee plots. It was shown that the V value of SHR was lower than that of WKY significantly. On the other hand, apparent Km was not different between SHR and WKY. In order to investigate the effect of parathyroid hormone (PTH) on Pi and D-glucose transport in SHR, parathyroidectomy was performed. Pi and D-glucose uptakes by BBMV prepared from SHR with PTX were also lower than that prepared from WKY with PTX. Phosphate, D-glucose, and L-alanine are all reabsorbed by Na-gradient-dependent transport system in renal proximal tubulus. Sodium ion permeability of BBMV prepared from SHR was also increased significantly. In this study, Pi, D-glucose, and L-alanine transport, which performed by Na-gradient-dependent transport system, were all decreased in SHR compared with WKY. In the absence of PTH, these phenomenon were not changed. CAMP dependent phosphorylation of BBMV was not changed between SHR and WKY. Kinetic study of Pi transport showed the change of capacity of transport system. From these results, we hypothesized that the stimulation of sodium ion permeability recognized in SHR caused the decrement of sodium gradient, and this decrement effected Na-gradient-dependent transport system.

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