Urinary Recovery of Salicylamide as Its Glucuronide in Liver Disease

An attempt was made to establish the method for the estimation of glucuronide formation in vivo using salicylamide and further to study the alteration in the glucuronide formation in liver disease. The results were as follows: 1. The method for the determination of free salicylamide separately from other conjugates of salicylamide in urine, without involving any hydrolysis of the other conjugates, was presented. When 1 g. of salicylamide was administered to the subjects with or without liver injuries, no free salicylamide was detected by the present method in the urine following the salicylamide administration. 2. The analytical method for the determination of salicylamide glucuronide was also devised by employing a hydrolysis with &#946;-glucuronidase. The ratio of the salicylamide liberated by the enzymatic hydrolysis of the 10-hour urine following the administration of 1 g. of salicylamide to the total salicylamide excreted in the same urine was neither affected by the total recovery of the salicylamide nor by the urine volume. This ratio was thus used as a means of estimating the capacity of the glucuronide formation in vivo, although it was considered that the ratio might be affected to some extent by the competition between the glucuronide and other conjugate formations in vivo. 3. As a result of this salicylamide glucuronide excretion test, it was indicated that the in vivo formation of salicylamide glucuronide in the patients with postnecrotic cirrhosis was slightly decreased compared with that in normal controls.</p

Enzymatic Studies of Glucuronide Formation in Impaired Liver III. Effects of Carbon Tetrachloride and Ectromelia Virus Infection on Liver Glucuronide Formation in Mouse

Glucuronide formations by mouse liver homogenate in several liver impairments were studied by using 4-methyl umbelliferone as a glucuronide receptor. The results were as follows : 1. Subcutaneous or intraperitoneal administration of carbon tetrachloride to the mouse produced a significant increase in the liver glucuronyl transferase activity 12 or 24 hours after the treatment regardless of histological and enzymatic evidences of liver-cell necrosis. This increase was not attributed to the increase in the 'activator' of glucuronide formation but to the increase in the enzyme activity itself. 2. In Ectromelia virus mouse hepatitis, the glucuronyl transferase activity of the liver tissue was markedly reduced in severe cases. In moderate or milder cases, a slight increase in the activity was observed in a few of them in the early stage of the disease, and the activity was significantly decreased on the recovery in all of the cases which survived. 3. In the early stage of carbon tetrachloride injury when the glucuronyl transferase activity of whole mouse liver was increased and the decomposition of uridine diphosphate glucuronic acid by the liver tissue was also enhanced, the glucuronide formation in vivo was rather increased. It was thus considered that the whole liver glucuronyl transferase activity rather than the uridine diphosphate glucuronic acid content was responsible for the glucuronide formation in vivo as a rate-limiting factor.</p

Enzymatic studies of glucuronide formation in impaired liver. II. Activating effect of boiled liver extract on liver glucuronide formation

The effect of the boiled liver extract on the velocity of 4-methyl umbelliferone lucuronide formation by mouse liver homogenate was studied. The results were as follows: 1. Addition of mouse or rat liver boiled extract to the complete system for the glucuronide formation produced an increase in the velocity of the glucuronide formation. 2. The boiled liver extract produced the increase in the velocity of the glucuronide formation not as a substrate but as an activator. 3, The activator in the boiled liver extract was relatively heat stable, acid labile, and precipitated as a fairly ethanol-soluble barium salt. The solution of the activator partially purified by ethanol fractionation of the barium salt indicated its absorption maximum at 262 m&#956;. These results suggested that the most possible substance in the boiled liver extract responsible for the activation of 4methyl umbelliferone glucuronide formation might be a sugar nucleotide.</p

Isolation of Endogenous Activator of Glucuronide For­mation in Liver

A purified compound of the activator of glucuronide formation was isolated from a boiled extract of rat liver by charcoal adsorption, ethanol fractionation of barium salts, and finally paper and Dowex-l column chromatographies. The analytical data and the chemical properties of the compound sugggested that the endogenous activator of glucuronide formation in rat liver might be uridine diphosphate N-acetylglucosamine.</p

Enzymatic Studies of Glucuronide Formation in Impaired Liver I. Assay Methods For the Determination of Glucuronyl Transferase Activity and Uridine Diphosphate Glucuronic Acid Content of Liver Tissue Using 4-Methyl Umbelliferone as a Glucuronide Receptor; Its Application to Needle Liver Biopsy Tissues

An attempt was made to apply the method devised by ARIAS for the determination of liver glucuronyl transferase activity using 4-methyl umbelliferone as a glucuronide receptor to the small amounts of liver tissue obtained by needle biopsy. This was accomplished by studying the kinetics of enzymatic 4-methyl umbelliferone glucuronide formation by mean(of mouse liver homogenates. The improved method was proved to be applicable to human liver and gave a satisfactory result. In addition, an assay method for the estimation of liver uridine diphosphate glucuronic acid content from the amount of 4-methyl umbelliferone glucuronide formed from the uridine diphosphate glucuronic acid contained in the liver homogenate used as a source of glucuronyl transferase was studied, and as a result it was proved to be also applicable to the small amounts of human liver tissue.</p

Effect of vitamin B12 derivatives on urinary excretion of methylmalonic acid in liver diseases

1. Twenty.one patients with liver diseases were studied for their urinary mehylmalonic acid excretion after a valine load by means of an improved thin layer chromatography. 2. Methylmalonic acid positive cases were found in four out of the ten patients with cirrhosis of the liver, all four with cirrhosis and diabetes mellitus, and none with acute hepatitis of icteric phase. No apparent correlation was found between the methylmalonic acid excretion and the extent of hepatic damage. 3. A large amount of methylmalonic acid found in the case (S. I.) with cirrhosis of the liver and diabetes mellitus after the valine load was not corrected by cyanocobalamin but by DBCC, suggesting an impaired transformation from cyanocobalamin to DBCC. However, the nature of the impairment remains unknown.</p

Effect of gluconeogenic diet on primary hepatoma patients

Three inoperable patients with primary hepatoma could be placed on gluconeogenic diets (minimum carbohydrate-high fat diets) for one to three months. A transient inhibition or a marked retardation of the tumor growth was observed with these patients and their entire clinical courses were fairly good. These results confirmed our previous observation with a metastatic liver tumor patient.</p

Treatment of hepatic cancer patients with a gluconeogenic diet: report of a case with hepatic metastases of malignant melanoma

A case of malignant melanoma with metastases mainly to the liver and the right ilium was treated with a gluconeogenic diet. The carbohydrate content of the diet was finally reduced to 5&#8764;10 g per day and the remaining calories were derived from protein and fat. Increased blood citrate and NEFA concentrations, increased ketone body formation and the maintenance of a reasonable level of blood sugar confirmed the attainment of a gluconeogenic metabolic state. Definite improvements in size of a hepatic tumor, serum alkaline phosphatase activity and the general condition were observed transient1y during the dietary therapy. Growth of the tumor resumed despite the continued gluconeogenic therapy, and the patient died of cardiac failure. Concentrations of gluconeogenic enzymes, fructose-1, 6-diphosphatase, glutamic oxaloacetic transaminase and glutamic pyruvic transaminase, were all found to be very low in the tumor tissue as expected.</p

Wider Distribution of Salivary-Type Isoamylase Activity as Compared with Pancreatic-Type Isoamylase Activity in Serum: A Study on Young Female Adults

The clinical implications of a wider distribution of salivary type (S-type) isoamylase activity, as compared with that of pancreatic type (P-type) isoamylase activity in the serum of young female adults of 18-23 years old was studied. A high correlation existed between the S-type isoamylase levels in the initial determination and those in the second determination one year after on the same subjects, indicating that the wider distribution of S-type isoamylase level reflects an individual variation. The serum level of S-type isoamylase was highly correlated with the S-type isoamylase activity in saliva. Among the additional factors studied, a weak positive correlation was present between energy intake and the total and S-type isoamylase activities in serum. However, there was no negative correlation between the S-type isoamylase level and body mass index (BMI), which was reported for young male adults