An investigation of 2,093 renal biopsies performed at Tokai University Hospital between 1976 and 2000.

We retrospectively investigated 2,093 renal biopsy procedures performed between 1976 and 2000 at Tokai University Hospital. The study period was divided into 5-year intervals, and the frequencies of each renal disease, age and sex of patients were compared across the study period. Clinical diagnosis was based on WHO criteria. A total of 2,093 patients aged 8 months to 84 years underwent renal biopsy during the study period. The percentage of elderly patients who underwent renal biopsy increased from 1.2% in 1976 - 1980 to 9.9% in 1996 - 2000. IgAN was the most common disease in every 5-year period. CresGN showed an increase from 1 patient (0.3%) in 1976 - 1980 to 15 patients (4.0%) in 1996 - 2000. In contrast, the number of patients with PGN or BRH, MPGN significantly decreased during the study period. Although the criteria for renal biopsy and renal diseases detected are expected to change in the future, renal biopsy will remain an essential procedure for making a definite diagnosis, selection of optimum treatment, and prediction of prognosis.

In situ hybridization of interleukin 6 in diabetic nephropathy.

Increased mesangial expansion is one of the most characteristic histological changes in diabetic nephropathy (DN). Although the pathogenesis of DN remains unclear, recent studies associate interleukin (IL) 6 with mesangial proliferative glomerulonephritis. To elucidate the expression and localization of IL-6 mRNA in renal tissues of patients with DN, a high-resolution in situ hybridization using digoxigenin-labeled oligonucleotide was performed. Patients were divided into three groups based on light microscopy findings: mild (group 1), moderate (group 2), and severe (group 3) mesangial expansion. The relationship between the expression of IL-6 mRNA and the degree of glomerular mesangial expansion in DN was examined. Individual cells positive for IL-6 mRNA were observed in glomeruli. These cells were mesangial cells, glomerular epithelial cells, and Bowman's capsule. The signal intensity was strongest in tissues from group 2 but was weak in those from groups 1 and 3. Most cells in the area of mesangial proliferation were strongly stained for IL-6 mRNA, and few positive cells were found in the Kimmelstiel-Wilson nodular lesion. In the interstitium, some tubules, particularly atrophic tubules, and some infiltrating cells were positively stained for IL-6 mRNA. The interstitial expression of IL-6 mRNA correlated significantly with the degree of interstitial injury and was remarkable in tissues from groups 2 and 3. We conclude that IL-6 mRNA is expressed by glomerular resident cells and interstitial cells in the renal tissue of patients with DN and that its expression may be associated with mesangial proliferation and may be involved in the tissue injury of DN.

Expression of 5-lipoxygenase and 5-lipoxygenase activation protein in glomerulonephritis.

The gene transcription of 5-lipoxygenase (5-LO) and 5-lipoxygenase-activating protein (FLAP) in renal tissue of patients with IgA nephropathy and mesangial proliferative glomerulonephritis were analyzed by amplification of reverse transcribed mRNA with polymerase chain reaction (PCR). The level of expression was determined by analysis of the PCR products of 5-LO, FLAP, and beta-actin. Forty percent of the patients expressed both 5-LO and FLAP. Relative to beta-actin, FLAP expression (0.42 +/- 0.21) was higher than 5-LO (0.14 +/- 0.10). Comparison of clinical data showed that patients who expressed 5-LO and FLAP had a lower glomerular filtration rate and an increased level of blood urea nitrogen, serum creatinine, and proteinuria. The results suggest arachidonic acid metabolism by inflammatory cells in renal tissue may play an important role in human glomerulonephritis.

Immunofluorescence staining of renal biopsy samples in patients with diabetic nephropathy in non-insulin-dependent diabetes mellitus using monoclonal antibody to reduced glycated lysine.

This is the first report on immunofluorescence staining of renal biopsy samples in human diabetic nephropathy (DN) using monoclonal antibodies to reduced glycated lysine. In order to detect the localization of glycated lysine in the mesangial matrix and/or the glomerular basement membrane (GBM), we examined immunofluorescence staining using antibodies against reduced glycated lysine in the glomeruli of 16 patients with DN and ten age-matched patients with diffuse mesangial proliferative glomerulonephritis without IgA deposition (DPGN) as controls. In the early stage of DN, immunofluorescence microscopy revealed the presence of intense staining for reduced glycated lysine in the GBM as well as in part of the tubular basement membrane, but not in the mesangial area. In contrast, immunofluorescence microscopy revealed less staining for glycated lysine in the GBM in the advanced stage of DN, and no reaction with any part of the renal tissue in patients with DPGN. It was concluded that detection of reduced glycated lysine in GBM in the early stage of DN might be associated with the initial pathogenesis of this disease.

Significance of high levels of serum IgA and IgA-class circulating immune complexes (IgA-CIC) in patients with non-insulin-dependent diabetes mellitus.

Significance of serum IgA and IgA-class circulating immune complexes (IgA-CIC) elevation in patients with non-insulin-dependent diabetes mellitus (NIDDM) was described. Seventeen patients with NIDDM and 17 patients with diffuse mesangial proliferative glomerulonephritis without deposition of IgA (DPGN) as controls were examined. The levels of serum IgA in patients with NIDDM were significantly higher than those in patients with DPGN (p < or = 0.01). The levels of IgA-CIC in patients with NIDDM were also significantly higher than those in patients with DPGN (p < or = 0.01). Production of IgA derived from B cells and the proportion of IgA bearing B cells in patients with NIDDM were not significantly higher than those in patients with DPGN. Furthermore, the levels of IgA in pharyngeal washings from diabetic patients were not significantly higher than those for DPGN patients. Duration of diabetes, the level of HbA1c, and the presence of hypertension, microalbuminuria, or retinopathy showed no significant correlations with the levels of serum IgA or IgA-CIC in patients with NIDDM. It was postulated that the elevations of serum IgA and IgA-CIC were based on subclinical infection of the mucosa and/or deterioration of IgA clearance in patients with NIDDM.

Preliminary study on specificity of IgA released from lymphocytes by EB virus transformation in patients with IgA nephropathy.

A study on the specificity of IgA in the supernatant of Epstein-Barr Virus (EBV) transformed lymphocytes obtained from patients with IgA nephropathy is described. Renal biopsy specimens were obtained from nine patients with IgA nephropathy and nine patients with other glomerular diseases. Mononuclear cells obtained from two patients with IgA nephropathy and two healthy adults were transformed with EBV and the supernatant of such transformed cell culture was applied to acid (pH 3.2)-treated renal sections obtained from the same and other patients with IgA nephropathy, as well as to those with other glomerular diseases. The sections were stained with FITC-labeled heavy chain-specific anti-human IgA antiserum and then examined with a fluorescent microscope. It was demonstrated that IgA in the supernatant specifically bound with the glomerular mesangial areas in patients with IgA nephropathy, while it did not bind with such areas in patients with other glomerular diseases. Although IgA in such samples bound with autologous renal tissues, about 50% bound with allogeneic renal specimens of IgA nephropathy. IgA in the supernatant of transformed lymphocytes from healthy adults was not bound with the glomerular mesangial areas of patients with IgA nephropathy and other glomerular diseases. It is concluded that IgA in the supernatant of lymphocytes by EB virus transformation is specific to the mesangial areas of IgA nephropathy and that some heterogeneity is seen among patients with this disease.

Evaluation of renal biopsy samples of patients with diabetic nephropathy.

OBJECTIVE: To evaluate the usefulness of renal biopsy in the overall management of patients with diabetes mellitus (DM), we examined the relationship between the clinical parameters and histopathological findings of renal biopsy samples. METHODS: Renal biopsy specimens were obtained from 109 type 2 diabetic patients with proteinuria. Samples were divided into the following two groups: Diabetic Nephropathy (DN) group (n=80) had typical diabetic lesions without other renal diseases, complication group (n=29) had diabetic lesions with other renal diseases. Furthermore, DN group was subdivided into two subgroups: slow progressive group (SP group, n=32), the level of serum creatinine (s-Cr) was normal at the time of renal biopsy and three years after renal biopsy, and fast progressive group (FP group, n=14), the level of s-Cr was normal at the time of renal biopsy but more than doubled three years after renal biopsy. RESULTS: The level of total protein was significantly lower and HbA1c significantly higher in the DN group than in the Complication group. However, other clinical parameters were not significantly different between the two groups. Urinary protein, systolic and diastolic blood pressure in FP group were significantly higher than in SP group. The percentage of sclerotic glomeruli, the severity of mesangial expansion, tubular injury and cell infiltration were significantly greater in FP than in SP group. CONCLUSIONS: Our results indicated that a complete evaluation of renal pathology in DM could not be made by clinical parameters only, and that the progression of DN could be accurately predicted by histopathological evaluation. Therefore, this study emphasizes the importance of renal biopsy in the overall management of patients with DM and/or DN.

A diabetic case with hemoglobin J-Meerut and low HbA1C levels.

A diabetic patient with hemoglobin (Hb) J-Meerut and low HbA1C levels is reported. An automatic glycohemoglobin analyzer used for the determination of HbA1C revealed an abnormal peak of the peripheral blood obtained from a Japanese female with diabetes. She showed a lower HbA1C level (3.7%) than expected from her fasting plasma glucose (172 mg/dl). High performance liquid chromatography and isoelectric focusing indicated that her abnormal hemoglobin was Hb J-Meerut [alpha 120(H3)Ala-->Glu] and it accounted for 28.3% of the total hemoglobin. Abnormal hemoglobinemia should be considered when a major discrepancy between the levels of HbA1C and fasting plasma glucose is observed.

Urinary albumin fragments as a new clinical parameter for the early detection of diabetic nephropathy.

Urinary albumin fragments (uAF) from patients with NIDDM were analyzed as a possible factor in the early discovery of diabetic nephropathy before emergence of microalbuminuria. SDS-PAGE and immunoblot assay were employed for detection of uAF. Samples from 252 patients with NIDDM, 158 patients with non-diabetic diseases, and 48 healthy volunteers were examined; uAF were detected in 139 (55.2%), 94 (59.5%), only one (2.1%), respectively. In diabetic patients with normoalbuminuria, uAF were detected in 48 out of 159 cases (30.2%). Two years after the initial study, 3 of the 17 diabetic patients (17.6%) with normoalbuminuria and uAF showed micro- or macroalbuminuria. It was concluded that detection of uAF might be useful for the early detection of diabetic nephropathy in NIDDM.

Significant correlation between the immunofluorescence of alpha 2-plasmin inhibitor in glomeruli and the effects of urokinase therapy in patients with IgA nephropathy.

Correlation between the deposition of alpha 2-plasmin inhibitor (alpha 2-PI), which is one of the inhibitory factors of fibrinolytic activities, in the glomeruli and the effects of urokinase therapy in patients with IgA nephropathy is described. Urokinase (UK) is a plasminogen activator derived from fresh human urine. Urinalysis and measurements of renal function tests, i.e., serum creatinine, blood urea nitrogen, glomerular filtration rate and phenolsulfonphtalein, were performed before and at 8 and 48 weeks after the administration of urokinase. There was marked improvement of proteinuria after UK therapy in patients without deposition of alpha 2-PI in the glomeruli. In contrast, the improvement of proteinuria after UK therapy was not observed in patients with positive deposition of alpha 2-PI in the glomeruli. It was concluded that the administration of UK may be useful for treatment of proteinuria in patients with IgA nephropathy.

Clinical effect of newly developed oral carbonaceous adsorbent (AST-120) in patients with advanced stages of chronic glomerulonephritis.

Clinical effects of a newly developed oral carbonaceous adsorbent (AST-120) in patients with advanced stages of chronic glomerulonephritis is described. Measurements of serum creatinine were performed before, during and after the administration of AST-120. The reciprocal of serum creatinine concentration was also plotted serially in each patient. The mean levels of the slope of the reciprocal serum creatinine vs time plot after administration of AST-120 was decreased compared with those before the administration of AST-120 but there was no statistical significance (p greater than 0.10). Further studies with more patients are needed to determine whether the administration of AST-120 may be useful for treatment of patients with chronic glomerulonephritis.

[Two cases of metastatic renal tumor].

In this report, two cases of metastatic renal tumor are described. The first case is a 58-year-old female who had branchiogenic carcinoma. She had found a left neck tumor, before asymptomatic macrohematuria developed. The second case is a 25-year-old female who had choriocarcinoma. She had no urological symptoms. In both cases, unilateral nephrectomy was performed. Seventy nine cases of metastatic renal tumor in survivals have already been reported.

Molecular weights of IgA-circulating immune complexes (CIC) in patients with IgA nephropathy: application of HPLC and solid-phase anti-C3 Facb EIA.

The detection of molecular weights of IgA-circulating immune complexes (CIC) in sera is described. Measurement of IgA-, IgG- and IgM-CIC in sera was also performed. Serum samples were obtained from nine patients with IgA nephropathy, nine patients with chronic proliferative glomerulonephritis without IgA deposits (PGN) and 19 healthy adults. Serum samples of these patients were subjected to high performance liquid chromatography (HPLC) and then divided into 50 fractions. Measurement of IgA-, IgG- and IgM-CIC in such fractions was performed using solid-phase anti-C3 Facb EIA. The levels of IgA-CIC in sera from patients with IgA nephropathy were significantly higher than those from patients with PGN or healthy adults. The peak values of molecular weights of IgA-CIC in sera from patients with IgA nephropathy ranged from 2.6 X 10(5) to 3.0 X 10(5) daltons (mean +/- SD; 2.9 +/- 2.0). It was suggested that the increase of IgA-CIC in sera from patients with IgA nephropathy may be mainly due to dimers and/or polymers.

Clinical effect of dipyridamole in patients with IgA nephropathy.

Clinical effects of dipyridamole and carbazochrome sodium sulfonate in patients with IgA nephropathy are described. Oral administration of 300 mg of dipyridamole and 180 mg of carbazochrome sodium sulfonate per day was employed in the present study. Urinalysis and renal function tests, i.e. serum creatinine (s-Cr), blood urea nitrogen (BUN), glomerular filtration rate (GFR) and phenolsulfonphtalein (PSP) tests, were performed before and after the administration of dipyridamole. It was demonstrated that the administration of dipyridamole was effective in reducing the level of proteinuria in the patients. The administration of carbazochrome sodium sulfonate was not effective in reducing the proteinuria level. It was concluded that the administration of dipyridamole may be useful for treatment of patients with IgA nephropathy.

Levels of fibrinopeptide A (FPA) and fibrinopeptide B beta 15-42 (FPB beta) in urine and plasma after urokinase administration in patients with chronic glomerulonephritis.

A study on the significance of the measurement of fibrinopeptide A (FPA) and fibrinopeptide B beta 15-42 (FPB beta) in urine and plasma before and after urokinase (UK) administration in patients with chronic glomerulonephritis is described. FPA and FPB beta in urine and plasma were measured by radioimmunoassay. The levels of fibrinogen degradation product (FDP) in urine and sera were also determined before and after the UK administration. UK was administered at a dose of 48,000 IU/20 ml in saline intravenously, followed by an intravenous infusion of 250 ml of 5% glucose for one hour. It was demonstrated that the levels of FPB beta in urine and plasma showed a significant increase after UK administration in patients with chronic glomerulonephritis. However, there was no significant difference in the levels of FDP in urine and sera before and after UK administration. It was concluded that the measurement of FPB beta in urine and plasma may be useful for evaluating the efficacy of UK therapy in patients with chronic glomerulonephritis.

An adult case of Fanconi's syndrome associated with membranous nephropathy.

An adult case of Fanconi's syndrome associated with membranous nephropathy is described. Renal biopsy specimens revealed typical features of membranous nephropathy associated with severe tubulo-interstitial changes in light microscopy, electron microscopy and immunofluorescent microscopy. The nephrotic syndrome due to membranous nephropathy was observed at the time of definite diagnosis of Fanconi's syndrome in this patient. It is suggested that renal tubular injuries in some patients with membranous nephropathy might be an important factor in the pathogenesis and/or development of Fanconi's syndrome.

Detection of alpha 2-pregnancy associated-glycoprotein (alpha 2-PAG) in cultured human peripheral blood mononuclear cells.

A study of the detection of alpha 2-pregnancy associated-glycoprotein (alpha 2-PAG) by double immunofluorescence in cultured human peripheral blood mononuclear cells (MNC) was performed. MNC was obtained from four healthy adults and isolated on a Ficoll-Hypaque density gradient. MNC was incubated with RPMI-1640 with or without pokeweed mitogen in 5% CO2/air at 37 degrees C for six days. MNC was initially stained with rhodamine-conjugated antihuman IgA, IgM, and IgG antisera, and then stained with FITC-conjugated antihuman alpha 2-PAG antisera. It was concluded that alpha 2-PAG was not produced by IgA, or by IgM and IgG producing cells in healthy adults.

Siblings with IgA nephropathy and diffuse proliferative glomerulonephritis (PGN) associated with identical HL-A antigens.

Siblings with IgA nephropathy and diffuse proliferative glomerulonephritis without mesangial IgA deposits (PGN) who had identical HL-A antigens are described. A 21-year-old woman suffered from IgA nephropathy and her 27-year-old sister showed diffuse proliferative glomerulonephritis (PGN) distinct from IgA nephropathy. These siblings had identical HL-A alloantigens which were A2, Aw24; Bw16, Bw35; Cw4, Cw7; DR4. Microhematuria and increased serum IgA levels were shown in their father and other sister but renal biopsies were not performed. The serotypes for the HL-A antigens of their father and other sister were A2, Aw24; Bw27, Bw35; Cw4; DR4 and A2; Bw16; Cw7; Dr2, respectively. It is demonstrated that IgA nephropathy and PGN share a common HL-A phenotype including DR4. It is postulated that the presence of DR4 antigens is not directly related to the occurrence of IgA nephropathy.

Detection of immunoglobulins and/or complement in kidney tissues from non-obese diabetic (NOD) mice.

Immunofluorescent studies were performed on kidney tissues from non-obese diabetic (NOD) mice. Tissues from twenty-two NOD mice ranging in age from 8 to 33 weeks were examined by immunofluorescence and light microscopy. It was shown that the immunofluorescent pattern of immunoglobulins and complement deposition in glomeruli from NOD mice were changed from diffuse linear, focal and/or local fine granular to diffuse coarse granular depending on the age. These findings in NOD mice were consistent with those in human diabetic nephropathy. It was suggested that the immunofluorescent studies in NOD mice are useful for evaluation of the pathogenesis of renal involvement in patients with diabetes mellitus.

A case of mesangial IgM nephropathy with decreased renal function.

A case of mesangial IgM nephropathy associated with chronic renal failure is described. The patient did not reveal nephrotic syndrome during the clinical course. Renal biopsy specimens revealed typical features of mesangial IgM nephropathy when studied by light microscopy, electron microscopy and immunofluorescence staining. Mesangial IgM nephropathy is considered to be a heterogenous disorder, because this disorder is clinically characterized by nephrotic syndrome, mild proteinuria and/or hematuria or renal failure.