Further studies on an eleventh case of heavy (Hgamma1) chain disease--biosynthetic studies

In vitro quantitative biosynthetic studies were carried out on bone marrow cells obtained from an eleventh case with gamma heavy chain disease. The findings indicate that neither cytoplasmic nor extracellular degradation was responsible for the presence of the gamma heavy chain fragment in serum. The absence of a covalent-bound light chain was also confirmed.</p

Further studies on an eleventh case of heavy (Hgamma1) chain disease--physico-chemical studies

An abnormal protein with similar antigenic properties to Fc fragments of IgG, was found in the serum and urine of an eleventh case of heavy (Hgamma1) chain disease (Yok). This protein was purified with ammonium sulfate precipitation and by column chromatography of DEAE cellulose, CM cellulose and Sephadex G-200. The purity of the protein obtained was 98.5%. It was crystallized easily, forming thin hexagonal plates of various sizes. The chemical compositions and physical properties of the protein including viscosity, partial specific volume, diffusion constant, sedimentation constant, frictional ratio, extinction coefficient and iso-ionic point are reported.</p

Studies on bile pigments Part 1. 'Clinical significance of biliary bilirubin-phosphate fraction in various liver diseases

Bile was collected with a duodenal tube from 14 controls and 45 cases with various liver diseases which were consisted of 8 cases with acute hepatitis, 10 cases with chronic hepatitis, 8 cases with cirrhosis of the liver, 4 cases with Gilbert's syndrome, 3 cases with Dubin-Johnson's syndrome and 12 cases with cholecystopathy. Bilirubin was extracted from the bile and separated with Billing's siliconized Kieselguhr column chromatography. Bilirubin-phosphate fraction showing positive phosphate ester reaction reported by Kondo, was fractionated with cellulose column chromatography as reported by Monobe, and finally its percentage to total direct bilirubin was calculated. Correlations between bilirubin-phosphate fraction and various liver function tests were studied and the following results were obtained. 1) Bilirubin-phosphate fraction was found to be 7.3% in cirrhosis of the liver, 5.5% in constitutional jaundice, 5.0% in acute hepatitis, 3.5% in chronic hepatitis, 2.2% in cholecystopathy and 2.1% in controls. Percentage of bilirubin-phosphate fraction was significantly higher in cirrhosis of the liver than in control, cholecystopathy and chronic hepatitis, and in acute hepatitis than in control and cholecystopathy. 2) There were good correlations among percentage of bilirubin-phosphate fraction, colloidal reaction (ZnTT, TTT) and serum γ- globulin fraction. 3) No significant correlations could be found among percentage of bilirubin-phosphate fraction, serum transaminase (GOT, GPT), cholesterol and alkaline phosphatase. 4) There was a good correlation between percentage of bilirubin-phosphate fraction and BSP retention at 45 minutes. 5) A significant negative correlation was obtained between percentage of bilirubin-phosphate fraction and K(ICG)

Studies on bile pigments Part 2. Spectrophotometric studies and clinical significance of non bilirubin fraction with positive diazo reaction in bile

Non bilirubin fraction which was extracted from bile of various liver and biliary tract diseases and prepared from crystalline bilirubin after irradiation by fluorescence was studied. Following results about the nature of non bilirubin fraction positive for diazo reaction and its clinical significance was obtained. 1) The material obtained in the fraction was yellowish and soluble in water and showed positive diazo reaction. Its absorption maximum showed 545-555nm in water solution (pH 2.4) and unchanged by acidulation. 2) In thin layer chromatogram of this azo pigment of non bilirubin fraction, two to four spots appeared usually. This results suggested that it was not composed from one substance. The Rf values were different from those of already reported azo pigments in thin layer chromatogram and it was confirmed that the Rf value of one of these spots was changed by methylation. 3) Percentage of non bilirubin fraction in total bilirubin in bile of various liver and biliary tract diseases was significantly increased in comparison with that of control subjects. This ratio increased in the order of the group of constitutional jaundice, cirrhosis of the liver and chronic hepatitis respectively. 4) Significant correlations between percentage of non bilirubin fraction in total bilirubin and liver function tests (ZnTT, TTT, γ-globulin) were obtained. These findings suggested that non bilirubin fraction was increased in the case of hepatic parenchymal damage or the disturbance of the bilirubin conjugation enzyme

Absorptions spectra of indirect and direct bilirubin

Direct bilirubins in vivo was fractionated into bilirubin diglucuronide, bilirubin phosphate and bilirubin sulfate. Influence of pH concentration on the absorptions maximum of each azopigment was studied. The schift of absorptions maximum was the same in the former two direct bilirubins and the latter moved as nearly as the indirect bilirubin-azo pigment yielded after an addition of methanol. These results was supposed to the difference was based on the structual difference of bilirubin due to the conjugation type of bilirubin

Absorptions spectra of indirect and direct bilirubin

Direct bilirubins in vivo was fractionated into bilirubin diglucuronide, bilirubin phosphate and bilirubin sulfate. Influence of pH concentration on the absorptions maximum of each azopigment was studied. The schift of absorptions maximum was the same in the former two direct bilirubins and the latter moved as nearly as the indirect bilirubin-azo pigment yielded after an addition of methanol. These results was supposed to the difference was based on the structual difference of bilirubin due to the conjugation type of bilirubin

Studies on non-bilirubin fraction showing positive diazo-reaction in heterozygote Gunn rat bile

Non-bilirubin fraction was extracted from the bile obtained from Wister strain rats and heterozygote Gunn rats, and separated by Ostrow(1)s method with minor modification. Azo-pigments was prepared with the non-bilirubin fraction after addition of Ehrlich(1)s diazo reagent. Significance of non-bilirubin fraction showing positive diazo reaction (NBAPF) was studied on the basis of the relation between the percentage of NBAPF to total bile azo pigments (proportion of NBAPF) and molar ratio of glucuronic acid to the ester-form bilirubin. The following results were obtained; 1) Averaged proportions of NBAPF were 0.94% in Wister strain rats and 2.50% in heterozygote Gunn rats, respectively. The latter proportion was higher than the former but not significant due to the wide variation of the measured values of the latter group. 2) Good correlation between the proportion of NBAPF and the molar ratio of glucuronic acid to the ester-form bilirubin was observed. 3) No significant correlation was observed between the bilirubin concentration in the bile and the molar ratio of glucuronic acid to the ester-form bilirubin or the proportion of NBAPF. 4) These results suggest when the conjugation of bilirubin and glucuronic acid in the liver was impaired, both the excretion of dipyrryl substance yielded from bilirubin and other conjugation mechanism increase compensatory

Acta Medica Okayama Further studies on an eleventh case of heavy (Hgamma1) chain disease-biosynthetic studies Chizuko Miyamoto-Sudo † Further studies on an eleventh case of heavy (Hgamma1) chain disease-biosynthetic studies * FURTJlER STUDIES ON AN ELEVEN

Abstract In vitro quantitative biosynthetic studies were carried out on bone marrow cells obtained from an eleventh case with gamma heavy chain disease. The findings indicate that neither cytoplasmic nor extracellular degradation was responsible for the presence of the gamma heavy chain fragment in serum. The absence of a covalent-bound light chain was also confirmed. Abstract: In vitro quantitative biosynthetic studies were carried out on bone marrow cells obtained from an eleventh case with gamma heavy chain disease. The findings indicate that neither cytoplasmic nor extracellular degradation was responsible for the presence of the gamma heavy chain fragment in serum. The absence of a covalent-bound light chain was also confirmed. The first case report of gamma heavy chain disease showed that extracellular catabolism appears not to be responsible for the existence of anomalous protein in patient serum (1). Several biosynthetic studies of the protein were carried out with peripheral blood lymphocytes and with bone marrow cells which showed the probable source of the labelled protein (2-4)