Toshio Okada

Immunohistochemical localization of glomerular basement membrane antigens in various renal diseases

The immunofluorescent localization of glomerular basement membrane (GBM) antigens was examined in 52 specimens from normal kidneys and in various renal diseases using antisera to human GBM HGBM), IV type collagen (IV Col) and P3 antigen, a rat nephritogen. Anti-HGBM serum normally stained the GBM and the mesangium in a restrictive pattern, anti-IV Col serum stained the GBM and the mesangium in a wider pattern and anti-P3 serum stained only the GBM. In mesangial proliferative glomerulonephritis, including IgA nephropathy pathy and Henoch-Schönlein nephritis, the widened mesangial areas were stained with anti-HGBM and anti-IV Col sera. In membranous nephropathy, the punched-out lesions of thickened GBM were demonstrated with the three antisera in moderate cases and a double linear distribution with fine granulation with anti-HGBM and anti-IV Col sera were revealed in one severe case. In membranoproliferative glomerulonephritis, the expanded mesangium and thickened capillary walls were stained with anti-HGBM and anti-IV Col sera, while the outer line of glomerular capillary walls was only positive with anti-P3 serum. In crescentic glomerulonephritis, the collapsed glomerular tufts were stained normally with anti-HGBM and anti-P3 sera and weakly with anti-IV Col serum. In diabetic nephropathy, anti-HGBM serum stained the GBM in a double linear distribution without reacting with the expanded mesangium; anti-IV Col serum stained the mesangium and the GBM in a less clear double linear fashion while anti-P3 serum stained the GBM as single line. Thin membrane disease and Alports syndrome had normal reactivity with all antisera. However, in one case of Alports syndrome anti-HGBM and anti-P3 sera stained the GBM in a focal and segmental pattern, while normal staining with anti-IV Col serum was found. In lesions with adhesions and crescents the staining was positive for HGBM and IV Col and negative for P3; obsolescent glomeruli were stained with anti-HGBM and anti-P3 sera, and had diminished staining with anti-IV Col serum. The identification of the various structural glomerular antigens is useful in the classification of certain types of glomerular diseases. Further insight into the mechanisms underlying these conditions may be obtained in this way.

Bronchial hyper-responsiveness to inhaled histamine in children with congenital heart disease

In order to assess bronchial responsiveness in patients with congestive heart failure secondary to congenital heart disease, we performed a histamine inhalation test while monitoring transcutaneous oxygen tension and compared the respiratory threshold to histamine with that obtained in patients with bronchial asthma. The inhalation test was performed by doubling concentrations of histamine solution for 2 min at 1 min intervals. The respiratory threshold of histamine was defined as the minimal concentration causing a drop in transcutaneous oxygen tension greater than 10% from baseline. Six of 10 patients with congenital heart disease and all of 12 patients with bronchial asthma had bronchial hyper‐responsiveness to histamine. The mean of histamine concentration was 2750μg/mL and 937μg/mL, respectively. During the histamine inhalation test, respiratory resistance gradually increased in congenital heart disease patients. This was measured by the linear slope of transcutaneous oxygen pressure (‐1.08 ± 0.75 mmHg/min), whereas in the bronchial asthma patients it rapidly decreased at the inflection point (‐4.19 ± 1.86 mmHg/min). We conclude that children with congestive heart failure had bronchial hyper‐responsiveness. We suggest bronchial hyper‐responsiveness to inhaled histamine in congestive heart failure was caused by the gradual increased respiratory resistance, which was different from that of bronchial asthma.

Organic Acid and Acylcarnitine Profiles of Glutaric Aciduria Type I

Urinary organic acid and acylcamitine profiles from a 2‐month‐old boy were studied by gas chromatography‐mass spectrometry and fast atom bombardment mass spectrometry. The patient excreted large amounts of glutaric acid and significant amounts of 3‐hydroxyglutaric acid, glutaconic acid and glutarylcarnitine, and his serum glutaric acid level was markedly elevated. Thus he was chemically diagnosed as having glutaric aciduria type I (GAI). In addition to the above metabolities previously described in GAI, significantly increased excretion of 2‐ketoglutaric acid, succinic acid, adipic acid, adipylcarnitine, suberic acid and azelaic acid was found. 2‐Ketoadipic acid methylsuccinic acid and ethylmalonic acid were also detectable, suberylcarnitine was not increased, and dehydroadipylcarnitine was decreased in his urine. These results suggest that excess glutaryl‐CoA causes the competitive inhibition of the dehydrogenation of adipylCoA to dehydroadipyl‐CoA and results in an increase of adipic acid and adipylcarnitine and a decrease of dehydroadipylcamitine. It is also suggested that oxidative decarboxylation of 2‐ketoglutaric acid to succinyl‐CoA is inhibited by high levels of glutaryl‐CoA, and that the dehydrogenation of succinic acid to fumaric acid is inhibited owing to the increased glutaric acid derived from excess glutaryl‐CoA. These results indicate that gas chromatography‐mass spectrometry is the most appropriate and accurate method for the differential chemical diagnosis of GAI and glutaric aciduria type II.

Chlorambucil central nervous toxicity: a significant side effect of chlorambucil therapy in childhood nephrotic syndrome

Chlorambucil (CHL) was used in combination with prednisolone in the treatment of nine children with frequently relapsing nephrotic syndrome. Serial electroencephalograms were obtained to evaluated CHL central nervous toxicity, before, during and after treatment with this agent. EEG abnormalities were observed in two of the nine children during chlorambucil therapy. EEG changes were diffuse spike and wave complexes and disappeared after discontinuation of therapy. There were no other neurological abnormalities and more particularly, no seizures or myocloni were observed. According to the literature, chlorambucil central nervous toxicity is found almost exclusively in childhood nephrotic syndrome. Strict neurological supervision of patients treated with chlorambucil is recomended.

Chromatographic profiles of urinary isoenzymes in healthy children

Urinary excretion of alkaline phosphatase, gamma-glutamyltransferase and lactate dehydrogenase was studied in a carefully selected group of 155 healthy children, 83 females and 72 males. Enzyme activity was assayed in randomly collected urine samples after gel filtration of the urine specimens. On chromatograms, urinary enzymes of alkaline phosphatase, gamma-glutamyltransferase and lactate dehydrogenase were separated into 4, 2 and 5 isoenzymes, respectively. Mean values of alkaline phosphatase, gamma-glutamyltransferase and lactate dehydrogenase activity were 4.59, 21.6 and 10.0 U/g creatinine. There were no sex-related differences besides lactate dehydrogenase which showed a higher excretion in females than in males. The excretion of urinary enzymes clearly decreased with increasing age.

The Low Molecular Weight of Protein Components in Children Urine

This study regards the urinary protein of patients with renal diseases. Analysis was done by micro‐vertical‐electrophoresis using polyacrylamide gel with sodium dodecyl sulfate (SDS‐PAGE). This method was an easy, reliable and rapid method for the estimation of low molecular weight proteinuria, which are present in renal tubular dysfunction. We found 6 patients with renal tubular dysfunction in 214 patients with renal diseases by the use of this method. Thus, the present method is considered to be useful for mass‐screening of children with proteinuria.

Serum Tumor Necrosis Factor in Mesangial IgA Glomerulonephntis with Macroscopic Hematuria in Children

Tumor necrosis factor (TNF)-alpha and interferon (INF)-gamma levels were measured in the sera obtained from 29 patients with IgA glomerulonephritis (IgA GN), 8 patients with minimal change nephrotic syndrome (MCNS) and 12 patients with upper respiratory tract infection (URI) without renal diseases in children. The serum TNF-alpha level of IgA GN was 123.0 +/- 175.4 pg/ml, MCNS was 4.9 +/- 4.0 pg/ml and URI was 10.5 +/- 4.5 pg/ml respectively. The serum TNF-alpha level of IgA GN was significantly higher than those of MCNS and URI. The serum TNF-alpha level of URI was on the high trend compared with that of MCNS, but was not statistically significant. Although the TNF-alpha level was related to mesangial cell proliferation in patients with IgA GN, it was unrelated to the grade of mesangial matrix expansion and magnitude of proteinuria. In 17 patients with IgA GN having macroscopic hematuria, the serum TNF-alpha level was 190.5 +/- 201.6 pg/ml, and in other IgA GN patients with microscopic hematuria it was 37.4 +/- 75.7 pg/ml. The serum TNF-alpha level of IgA GN with macroscopic hematuria was significantly higher than that with microscopic hematuria. In 6 patients with IgA GN with macroscopic hematuria, the serum TNF-alpha level was significantly decreased after macroscopic hematuria disappeared. The mean serum IFN-gamma level of IgA GN was 0.3 +/- 0.6 IU/ml, and MCNS was not detectable. Although the serum IFN-gamma level was related to mesangial cell proliferation in patients with IgA GN, it was unrelated to magnitude of proteinuria, the grade of mesangial matrix expansion and also the presence or absence of macroscopic hematuria. We suggest that macroscopic hematuria of IgA GN was closely related to the serum TNF-alpha level.

Automated separation and measurement of urinary isoenzymes and protein by ion-exchange liquid chromatography

This paper deals with a totally automated detection system for the assay of urinary isoenzymes and protein using high-performance liquid chromatography with a continuous post-column detection system. We attempted to determine the distribution of three enzymes in urine samples from a healthy child and in tissue extracts of rabbits. Alkaline phosphatase, gamma-glutamyltransferase and lactate dehydrogenase isoenzymes were each separated into six peaks. In comparison with the previous methods, this procedure provides better precision and accuracy, and it is sufficiently sensitive to allow the analysis without preconcentration of urine samples.

Endocardial fibroelastosis in two siblings with the infantile type and the childhood type

was 100 ml. No changes were observed in the levels of haemoglobin, blood sugar, liver function tests or electrolytes. Changes in pH and bicarboate levels are shown in Fig. 1. The pH fell rapidly after induction. It remained in the lower range and returned to preoperative levels within 72 h. One patient required intravenous administration of sodium bicarbonate to correct the rapid fall of pH. We have not been able to find a specific cause for the metabolic acidosis during anaesthesia of these patients. A carefut search of the literature did not reveal similar observations. Non-malignant hyperthermia that has occasionally been noted in patients with OI [2] was not present in any of our patients. None had hypoxia, hypercarbia or hypotension. Though many OI patients have reduced pulmonary function the ventilation-perfusion ratio is normal [1]. A controlled study is presently being performed in our institution.

A Case of Pelvic Osteomyelitis

A patient presenting with osteomyelitis of the pelvis is described. In this case it was difficult to establish a correct diagnosis by use of scintigraphic scanning, in spite of clear roentgenographic evidence of osteomyelitis.