In order to clarify the mechanisms of the initiation and progression of focal glomerular sclerosis (FGS), we investigated changes in the mesangial function or qualitative changes in the extracellular matrix of mesangium in puromycin aminonucleoside (PAN) -induced FGS in rats. At first, we investigated that the relationship between the progression of FGS and mesangial function. In order to evaluate the mesangial function, rats received the i. v. injection of colloidal carbon (C. C.) (20 or 30 mg/100 g). Results obtained from this experiment suggest that the progression of glomerular sclerosis may be related to changes in mesangial function. Furthermore, results suggest that the abnormality of the extracellular matrix may lead to changes in mesangial function and the progression of glomerular sclerosis. Therfore, in the next experiment, the proteoglycans (PGs), one of the components of extracellular matrix, were analyzed by the column chromatography to clarify qualitative changes in the PGs such as the molecular size and charge density. Results obtained from this experiment indicate that the sclerotic glomeruli synthesize the PGs with different molecular size and charge density from normal glomeruli. It is concluded from these experiments that the abnormality of the mesangial function and the synthesis of PGs, the components of the extracellular matrix, may lead to the progression of FGS. Namely, the qualitatively altered PGs may cause abnormal interactions with other components of matrix, which lead to changes in mesangial function, death of mesangial cell and the progression of FGS.

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In this paper, we studied the effect of elastase on aminonucleoside (AN) nephrosis which is considered a model of focal glomerular sclerosis (FGS). Elastase is an enzyme which disintegrates elastin, discovered by Balo, and used in the treatment of arterio-sclerosis and hyperlipidemia. It has also been known to improve metabolism of acid mucopolysaccharides, so, this study focused on metabolic improvement. Three groups of male Sprague-Dawley rats were studied and obsereved at regular intervals; 30, 60, 90 days. The ANE group (AN+elastase) was administered one shot of AN (10 mg/100 g B. W.) during the test interval, while elastase (5 mg/kg B. W.) was injected 5 days/week for the entire test interval. The AN group was administered one shot of AN only. The third group was a control (C). The following results were observed : (1) Focal segmental hyalinosis and sclerosis (FSHS), ANE group was weaker than AN group. (2) Other significant qualifying glomerular changes (vacuolar change and hyaline droplets of the epithelial cells, adhesion, and foam cells) ; ANE group was weaker than AN group. (3) Anion loss in GBM was shown by a lack of colloidal iron staining under light microscopy, and by a lack of PEI particles under electron microscopy; there was significantly less anion loss with ANE group, than with AN group. The findings suggest that elastase has an affect on the metabolism of acid mucopoly-saccharide and collagen in sclerotic lesion, and may restrain the progress of amino-nucleoside nephrosis.

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In an attempt to clarify the participations of cellular immunity in the development of tubulo-interstitial lesions, aberrant expressions of major histocompatibility complex (MHC) class II antigens and Ki-67 nuclear antigen on the renal tubular epithelial cells were studied. Ki-67 antigen was known to appear in all phases of cell cycle except for G_O. Nine normal kidney specimens (4 males and 5 females) and 117 kidney specimens obtained from patients with kidney diseases (54 males and 63 females) were examined with the indirect immunofluorescence technique using murine monoclonal antibodies against HLA-DR (la1), HLA-DQ (Leu10) and Ki-67 nuclear antigen. Patients included 100 with glomerular diseases, and 16 with tubulo-interstitial diseases consisting of 4 acute tubular necrosis (ATN), 7 acute tubulo-interstitial nephritis (AIN), one renal allograft rejection and 4 sarcoidosis. In normal kidney, HLA-DR was solely noted in only two specimens (22.2%) at the basal portion of proximal tubular epithelial cells. In tubulo-interstitial diseases 11(68.8%) out of 16 patients showed diffuse and intense expressions of HLA-DR concomitant with HLA-DQ in 6 of 13(42.9%), and 11 of 13(84.6%) were positive for Ki-67 nuclear antigen. Especially, in AIN and allograft rejection, intense expression of HLA-DR, DQ and Ki-67 nuclear antigen were observed in 100%, 86%, 100%, respectively. In ATN 3(75%) were positive for HLA-DR and Ki-67, but not for HLA-DQ. In contrast, only 12(15.6%), 2(2.6%) and 2(4.8%) of primary glomerular disease were weakly positive for HLA-DR, DQ and Ki-67 nuclear antigen, respectively. These results suggest that the aberrant expressions of HLA-DR and DQ in renal tubular epithelial cells in tubulo-interstitial diseases, especialliy in AIN and renal allograft rejection may play an important role in cellular immunity-mediated tissue injury, and expression of Ki-67 antigen is useful for in situ recognition of cellular activity and proliferation.

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We examined the clinical usefulness determined by polyacrylamide gel electrophoresis, followed by reaction with peroxidase-coupled lectins using urinary glycoproteins for diabetic nephropathy in 20 patients with diabetes mellitus. Lectins used were Triticum vulgaris (WGA), Phaseolus vulgaris (PHA-E₄), Dolichos biflorus (DBA), and Lens culinaris (LCA), which have high affinity for β1→4N-acetyl-D-gulcosamine (GlcNAcβ1→4GlcNAc), N-acetyl-D-galactosamine (GalNAc), a-galactosamine (α-GalNAc), and α-mannose (α-Man) residues, respectively. Electrophoretic patterns of urinary glycorproteins clearly showed the presence of lectin-reactive glycoproteins with molecular weights lower than that of albumin. The molecular weight of the main bands reacted with WGA, PHA-E₄ or LCA were 50, 000 and 38, 000, and increased with the progress of diabetic nephropathy. WGA reacted strongly with many glycoproteins having a wide range of molecular weights. LCA and PHA-E₄ reacted preferentially with glycoproteins of molecular weights glycoproteins of molecular weights lower than 50, 000, but no reaction was observed by DBA. These results suggest that low molecular urinary glycoproteins have abundant carbohydrate residues such as G1cNAc β1→4GlcNAc, GalNAc, and α-Man. The excretion of low molecular weight glycoproteins with high affinities for some lectins suggests functional impairment in diabetic nephropathy.

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The ultrastructural localization of immunoglobulins (IgG, IgA, IgM), complement component (C_3c), or fibrinogen-related antigen (FRA) was investigated on 5 biopsy samples from 5 children with idiopathic MN or lupus nephritis using proteinA-gold (PAG) complex method. The immunoreactivity of IgG was essentially confined to the mesangial and subepi-thelial electron-dense deposit (EDD) in all of 5 children, and more intense staining of IgG was observed in 3 children with stage II of MN than those with stage I and stage III of MN. Double immunocytochemical staining showed the same distribution of both IgG and IgM or both IgG and FRA in subepithelial EDD in 2 of 5 children. These findings suggest that IgG deposits are associated with the formation of subepi-thelial EDD in MN, and raise the possibility that IgM and FRA deposits may result from entrapment and/or immunological reaction.

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Certain uremic metabolites are recognized to have high affinity to serum protein and some of them have been identified as hippuric acid (HA), quinolinic acid (QA) and indoxyl sulfate (IS). Cell toxicity and effect on renal function of these substances were examined by means of erythroid colony formation, lymphocyte blast formation and isolated perfused rat kidney. These substances inhibited the binding of diphenylhydantoin to albumin, depending on the concentration of the substances. At the same concentration of 10 mg/dl, IS was most potent and QA was the second. QA and IS suppressed the erythroid colony formation, depending on the concentration of QA and IS. On the other hand, HA had no suppressive effect even at the higher concentration. The suppressive effect of QA and IS were attenuated by increasing erythropoietin concentration. QA and IS had strong suppressive effect of lymphocyte blast formation and interleukine 2 production at the concentration of uremic serum. However, they did not suppress the increase of intracellular calcium concentration of lymphocytes after stimulation by mitogen. This might indicate the possibility that these substances act not only on cell surface but on intracellular protein. HA and IS inhibited para-aminohippurate secretion and QA suppressed tubular react bsorption of sodium in isolated perfused rat kidney. These results show that the uremic protein binding inhibitors may influence renal regulation of fluid and electrolyte homeostasis. It is concluded that some of the protein binding inhibitors have toxic effects on cell function of various tissues and play a role in pathophysiology of uremia.

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In order to investigate the pathogenesis of hemolysis in chronic renal failure, red cell life span and hemolysis starting point (HSP) by coil planet centrifuge method were studied in 32 hemodialysis (HD) patients and 16 healthy subjects. Mean red cell life span was 22.0 days before recombinant human erythropoietin (r-HuEPO) therapy, and prolonged up to 28.1 days after r-HuEPO therapy. Mean HSP in HD patients was significantly elevated than healthy subjects. Mean HSP was 106.7 mOsm before r-HuEPO therapy, and improved to 101.0 mOsm after r-HuEPO therapy. HSP improved from 106.8 mOsm to 100.8 following the correction of bicarbonate level with HD. HSP was negatively correlated to bicarbonate level pre- and post-r-HuEPO therapy, but HSP was not correlated to BUN, serum creatinine, hydrogen ion, anion gap and amount of body water removal during HD. When blood PH in 18 HD patients was adjusted by 7% NaHCO₃ from 7.21 to 7.40, HSP improved to normal range in all. These data suggested that osmotic fragility was normal in younger erythrocyte in HD patients, and a decrease of plasma bicarbonate level resulted in an increase of hemolysis.

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The plasma level of Antithrombin-III (AT-III) antigen, antithrombin activity and the ratio of AT-III-heparin complex formation have been studied in 20 patients with chronic glomerulonephritis on regular hemodialysis at two occations with 2 year interval (in 1986 and 1988), and studied as to each variation in each plasma levels, and compared with the period of length of hemodialysis-history of individual cases. A significant decrease was found not only in plasma level of AT-III antigen (25.7+4.1 mg/dl to 23.4+3.9 mg/dl), but also in antithrombin activity (90.8±11.8% to 85.2±16.8%). Moreover and ratio of AT-III-heparin complex formation was significantly reduced during two years, too (50.7+3.0 to 53.0±1.5%). Similarly, a significant negative correlation could be found on the comparison between the duration of hemodialysis and each plasma levels of AT-III antigen (r=-0.27), antithrombin activity (r=-0.33), and as well as ratio of AT-III-heparin complex formation (r=-0.34). Therefore, it was interestingly suggested the possibility that prolonged hemodialysis treatment induced a functional loss of AT-III, especially in a capacity to form a AT-III-heparin complex due to partial loss of the affinity for heparin.

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Twenty cases of pre-eclamptic toxemia were examined clinicopathologically. Twelve primipara and 6 multipara underwent renal biopsy at 1-24(3.5±5.4, mean±SD) weeks after delivery. Specimens were examined by light and electron microscopy and immuno-fluorescence. Their mean blood pressure at delivery was 177.0±19.4/116.3±10.2 mmHg, proteinuria was 9.5±8.4 g/day. Follow-up period was 23.0±19.3 months after delivery. The severity of double contour of the glomerular basement membrane and mesangial interposition were correlated with the amount of proteinuria at biopsy (r=0.40, p<0.05, r=0.51, p<0.05, respectively). Proteinuria disappeared in all cases after delivery. The length of the hypertensive period after delivery correlated with the severity of glomerular lesions (r=0.63, p<0.05), but did not correlated with the severity of vascular lesions. All patients became normotensive within three months after delivery. No patients showed microhematuria during pregnancy or after delivery.

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78 patients with essential hypertension (17 with borderline hypertension and 61 with hypertension) and 13 normal controls were examined to evaluate the relation between the urinary excretion rate of guanidinoacetic acid/creatinine (U-GAA/Cr), β2-microglobulin/creatinine (U-BMG/Cr), radio-sensitive microalbumin excretion rate/creatinine (U-AER/Cr), N-acetyl-D-glucosaminidase/creatinine (U-NAG/Cr) and renal function. There was no significant difference among these groups in creatinine clearance (Ccr), serum creatinine (Cr) or in U-BMG/Cr, U-NAG/Cr and U-AER/Cr. In hypertensive patients U-GAA/Cr was 49.2±16.7 mg/gCr, which was much lower than in controls (78.1±13.4) (p<0.001). The Ccr has a significant relation with U-GAA/Cr (r=0.29, p<0.01) but not with U-AER/Cr, U-BMG/Cr nor U-NAG/Cr. In 44 patients, all of the above factors were investigated for 24 weeks during 4 kinds of anti-hypertensive treatment (10 with an angiotensin-converting enzyme inhibitor: A group, 11 with a β-adrenergic blocker : B group, 12 with a Ca entry blocker : C group and 12 with diuretics: D group). In A and C group, U-GAA/Cr was elevated during therapeutic course. However, in B and D group it declined during treatment. These findings suggested that urinary excretion of GAA may be a more sensitive marker than AER, BMG or NAG in hypertension and angiotensin-converting enzyme inhibitor and Ca entry blocker can be useful in the treatment of patients with essential hypertension with renal damage.

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The patient, a 28 year-old-man, was admitted to a hospital because of general fatigue and fever. He was pointed out renal dysfunction and was transfered to Nagasaki Uni-versity Hospital. The laboratory data on admission showed moderate azotemia (BUN 43 mg/dl, Cr 5.4 mg/dl). A percutaneous renal biopsy on admission revealed a diffuse crescentic glomerulonephritis. A direct immunofluorescence of renal biopsy showed a linear pattern for IgG along the glomerular basement membrane. Radioimmunoassay of his serum for circulationg anti-GBM antibody was strongly positive. Aggressive treatment with pulse therapy (methylprednisolone), plasmapheresis, and continuous heparin infusion was performed. He had markedly recovered from renal failure and escaped hemodialysis. The patient is making satisfactory process after 3 years.

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A case of atheroembolic renal failure diagnosed by renal biopsy was presented. A 69-year-old man was referred because of progressive renal failure two months after major arteiography for occlusive arterial disease of lower limbs. The physical examination on admission revealed an uncontrollable hypertension. The laboratory findings showed elevated serum creatinine (7.5 mg/dl) and eosinophilia (1022/mm³) with normal urinalysis findings. Renal biopsy disclosed a occlusive lesion of the arcuate artery which contained cholesterol clefts and foam cells, and showed ischemic renal parenchymal changes. These findings were compatible with cholesterol atheroembolic renal disease. In spite of the aggressive medical treatment, renal function had deteriorated progressively and the patient has been on regular hemodialysis. Atheroembolic renal failure after arteriography have been reported recently, but the case diagnosed by renal biopsy has been rare. Since there is no therapeutic way to reverse this type of renal failure, strict selection of patients for the angiographic examination and use of flexible catheter might be mandatory. Subacute course of renal failure after angiography and eosinophilia seem to be the important diagnostic clues for this disorder.

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A 28 year-old female patient who has been diagnosed as having systemic lupus ery-thematosus (SLE) developed an acute dissecting aneurysm of the aorta (DeBakey type I). The long-term, large dose corticosterid therapy (i. e., accumulative dose of about 60 g) administered for the treatment of lupus nephritis (WHO class III→IV) was considered to be responsible for a hypercholesterolemia (300-560 mg/dl) and a steroid-dependent hyper-tension (WHO class III) in this patient. The autopsy findings for the aorta were compatible with atherosclerotic changes but not with lupus arteritis. While atherosclerotic cardiovascular complications have been considered to be rare in patients with SLE, a growing body of evidence suggests that the incidence of such a complication may be increasing along with a dramatic improvement in the longevity of patients with SLE after an introduction of a large dose, long-term corticosteroid therapy.

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This paper reports two brothers with reflux nephropathy, Patients who were 10-year-old and 12-year-old were referred to our hospital due to proteinuria and deterioration of renal function. Diagnosis of reflux nephropathy was made on drip infusion pyelo-graply (DIP), voiding cystogram (VCG), and renal biopsy findings. The following findings were observed in renal tissues; focal and segmental sclerosis by light microscopy, IgM deposition by immunofluorescent microscopy, and glomerular basement membrane alterations and detachment of podocytes by electron microscopy, The HLA typing and analysis showed that both brothers and their mother possessed HLA-A9, which is reported to be closely associated with progession of primary reflux nephropathy to end-stage renal disease.

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The case was a 33-year-old woman with hypertension and hypokalemia, who presented depression of renin activity and the abnormal elevation of plasma deoxycorticosterone (DOC) and 11-deoxycortisol on laboratory tests. After admission, abdominal CT scan, 131I-adosterol scintigram and adrenal venogram revealed a tumor in the left adrenal, which histologically seemed to be benign. When the tumor was resected, blood pressure and all the biochemical data returned to normal range. DOC and ll-deoxycortisol levels in the tumor were abnormally elevated as compared with those in the normal adrenal tissue. These findings suggested that the abnormal elevation of hormone levels resulted from depression of 11β-hydroxylase. Though numerous adrenal tumors have been documented, we rarely encounter an apparently benign adrenal tumor that produces 2 kinds of hor-mones. This seemed to be the first case of benign adrenal tumor in which both DOC and ll-deoxycortisol were elevated.

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