The following tests using131I-labeled fat were performed on the patients with chronic pancreatitis who were admitted to our department from August in 1963 to December 1964. They all had a positive Kudo's test on P-point, clinical symptoms and abnormal endocrine and exocrine pancreatic functions. The results were compared with the control group having no disease of the digestive tract. 131I-oleic acid absorption test: The time required to reach the maximum level of radioactivity in the blood of 6 controls was 4 hours on the average. The average values were 11.4±1.8% and the values were between 9.5% and 14.6%. However, the time required to reach the maximum level of radioactivity in 12 pat ients with chronic pancreatitis was 5 hours on the average. The average values were 12.3±2.9%, and the values were between 9.9% and 19.1%. Since there was no significant differentiation between those with chronic pancreatitis and the controls, it is presumed that the function of absorption is normal. 131I-triolein absorption test: The time repuired to reach the maximum level of radioactivity in the blood of 12 controls is 4 hours on the average. The average values were14.9±1.6% and ranged from 14.1% to 17.4%. On 65 patients with chronic pancreatitis, the average time was 4 hours. The average values were 11.2±4.370, ranging from 12.0% to 21.9%. In some patients with chronic pancreatitis the absorption of 131I-triolein were moderately decreased. According to Ruffin et al, if the average of samples of the 4th, 5th and 6th hours are 8% or more of the administered dose of radioactivity, absorption is considered to have been normal. If the average of these 3 samples is less than 5%, it is assumed that marked disturbance of absorption has taken place. And if the averages werebetween 5% and 8%, the disturbance of absorption was moderate. In the present investigation, the mean of the 4th, 5th and 6th hours in controls was 14.6%, while the patients with chronic pancreatitis showed 11.1%. In th patients with chronic pancreatitis, 92% of them showed a marked !disturbance of absorption 23.1% of them showed a moderately disturbed absorption. The results of glucose tolerance tests did not appear in general parallel to the degree of disturbance of creatitis who reacted positively to Kudo's test on P-point, 32.3% also had a disturbance of the absorption of triolein as well as a greater than average sensitivity at the tender point. Howewer, the degree of tenderness and that of the disturbance of absorption of131I-triolein were not in general parallel.There has been much discussion in medical literature conserning digestion disturbance for chronic pancreatitis. In the present observation, digestive disturbance occured in 3 2.3% of chronic pancreatitis, but the 131I-oleic acid absorption disturbance was not observed in these patients.I-triolein absorption. Among the patients withchronic pan-creatitis who eacted positively to Kudo's test on P-point, 32.3% also had a disturbance of the absorption of triolein as well as a greater than average sensitivity at the tender point. Howewer, the degree of tenderness and that of the disturbance of absorption of 131I-triolein were not in general parallel. There has been much discussion in medical literature conserning digestion disturbance for chronic pancreatitis. In the present observation, digestive disturbance occured in 3 2.3% of chronic pancreatitis, but the 131I-oleic acid absorption disturbance was not observed in these patients.
The activities of urea cycle enzymes such as arginine synthetase (AS), ornithine transcarbamylase (OTC) and arginase (Arg.) were determined in the specimens obtained by liver needle biopsy carried out in 67 cases with various liver diseases, and at the same time, in 46 cases, the plasma levels of citrulline, arginine and blood ammonia were also determined for the purpose of clarifying the action of urea cycle in the liver in its pathological conditions and for studying the relationship between the activities of urea cycle enzymes and blood ammonia level. The following results were obtained. 1. The activities of liver urea cycle enzymes in normal controls (control liver specimens were obtained by resection on operating table at surgery for peptic ulcer from patients with normal liver function) were as follows: AS; 0.79±0.27, OTC; 59.8±15.1 and Arg.; 58.6±12.7μM/mg of soluble protein/60 min. The low level of the activity per mg of soluble protein in AS was fairly remarkable suggesting that the enzymological process synthesizing arginine from citrulline existed at the rate limiting step in urea cycle. The levels of plasma citrulline and arginine were 6.61±1.31 and 8.42±1.09μg/ml, respectively, and blood ammonia 0.66±0.24μg NH3-N/ml, 2. The activities of AS, OTC and Arg. in acute hepatitis were 0.39±0.32, 40.8±17.2 7. The activities of AS, OTC and Arg. in obstructive jaundice were 0.62±0.36, 49.1±13.7 and 42.8±18.9μM/mg of soluble protein/60 min., respectively.Decreased activities were seen in the cases with obstruction due to cancer, but in the cases with obstruction due to cholelithiasis no clear decrease in the activities was observed. The levels of plasma citrulline and arginine were 7.61±2.13 and 8.45±1.59μg/ml and blood ammonia 0.88±0.30μg NH3-N/ml.8. The activities of AS, OTC and Arg. in Banti's syndrome were 0.62±0.30, 54.3±19.4 and 49.6±11.6μM/mg. of soluble protein/60 min., respectively. No relationship was seen between the decrease of the activities of these enzymes and the onset time of this syndrome.and 41.6±21.3μM/mg of soluble protein /60 min., respectively. At the progressive or excerbating stage of acute hepatitis the activities were as follows: AS; 0.31±0.25, OTC; 39.1±20.1 and Arg; 30.6±13.3μM/mg of soluble protein /60 min. On the other hand, at the convalescent stage the activities of AS, OTC and Arg. were 0.53±0.41, 43.6±12.5 and 56.3±21.9μM/mg of soluble protein/60 min., respectively. Generally speaking, the decrease of the activities of these enzymes was nearly in direct proportion to the extent of necrosis of liver cells. The plasma levels of citrulline and arginine were 8.04±2.34 and 8.42±1.09μg/ml, respectively, and that of ammonia 0.75±0.31 μ g NH3-N/ml. The level of citrulline showed a tendency, though slight, to increase. 3. The activities of AS, OTC and Arg. in chronic hepatitis were 0.57±0.39, 51.4±18.9 and 45.8±22.4μM/mg of soluble protein/60 min., respectively, and the plasma levels of citrulline and arginine were 7.09±1.28 and 8.53±2.62μg/ml, respectively, and that of ammonia 0.83±0.23μg NH3-N/ml. 4. The activities of AS, OTC and Arg. in liver cirrhosis were 0.41+0.26, 42.9+12.6 and 43.9±11.9μM/mg of soluble protein/60 min., respectively. The decrease of the activities of these enzymes came next to that of acute hepatitis and a special decrease was observed in the cases at the non-compensatory stage of liver cirrhosis.
Leucine aminopeptidase, which is a proteolytic enzyme widely distributed in human tissues, serum and body fluids, has been studied utilizing several substrates. This report deals with a study of leucine aminopeptidase acting on L-leucylglycine (LG-LAP). Determination of its activity in serum and tissues was carried out with references to its isozyme patterns and activating or inhibitory effect of D-leucine upon the enzyme. Discussions were made on clinical usefulness of the serum LG-LAP activity in the hepatobiliary disorders, mechanism of increase in total LG-LAP and certain isozyme patterns and an activating effect of D-leucine in malignant diseases. Activity was measured by means of Fleisher's method with modified cololimetric method discribed by Yemm et al. Serum LG-I AP activity was determined in a total of 234 sera, obtained from 40 healthy persons and 146 patients, the majority of whom with various hepatobiliary disorders. Alteration of hydrolysis rate of LG-LAP by addition of D-leucine was investigated in 60 sera. Mobility of LG-I AP isozymes was measured by starch block electrophoresis in 7 sera. Tissue LG-LAP activity was assayed in 23 samples of the liver or stomach. The results are summarized as follows. 1) The highest level of serum LG-LAP activity was found in acute phase of viral hepatitis with subsequent slow decrease toward the normal during convalescence. Therefore, serial measurements of serum LG-LAP activity might be usefull in diagnosis and evaluation of prognosis of hepatitis. 2) Most of patients with primary or metastatic hepatic cancer and with obstructive jaundice due to malignant tumor showed modelately high serum LG-LAP levels. A subsequent high level was found in patients with cholangiolitic hepatitis, cholelithiasis and cholecystitis, but most of patients with chronic hepatitis cirrhosis of the liver and pancreatitis showed mild elevation. 3) Electrophoretic isozyme pattern of serum LG-LAP demonstrated five different.peaks in all instansis. In patients with acute hepatitis, the peak coincided withγ-globulin was enlarged. 4) Ten out of 16 patients with malignant tumor showed activation of LG-I AP by addition of D-leucine, while 29 out of 34 patients with benign disorders or normal subjects showed inhibition of LG-LAP by D-leucine. 5) LG-LAP activity in hepatic tissue was found to be decreased in patients with hepatitis and hepatic cirrhosis, while it was increased in patients with cancer, of the stomach. However, the activity in the cancerous tissue per se showed an increase in cancer of the stomach and a decrease in cancer of the liver. 6) It is postulated that increased serum LG-LAP in hepatobiliary disorders is mainly due to a release of enzyme from hepatic cells which are injured by parenchymal diseases but partially attributed to secondary hepato-cellular injury as a consequence of intra or extrahepatic biliary obstructions. 7) Observation on electrophoretic mobility and activation or inhibition by D-leucine suggest us that LG-LAP is composed of more than one enzyme proteins, but its details. are remained to be further studied.
Recent attentions in clinical enzymology have been paid toward on evaluation ofdiagnostic significance for alteration of blood enzymes activities as well as apathophysiologic analysis of their isozyme patterns. This report deals with a study of leucine aminopeptidase which acting on L-leucyl-β-naphthylamide (LN-LAP). Determination of its activity in serum, bile and tissue samples was carried out with referencesto its isozyme patterns and activating or inhibitory effect of D-leucine upon the enzyme. Discussions were made on clinical usefullness of the serum LN-LAP activity in thehepatobiliary diseases, mechanism of increase in total LN-LAP levels and certain isozyme patterns of serum and tissue samples, relationship between the serum and tissue LNLAP levels. A total of 347 sera from 40 normal subjects and 237 patients, majority of whom with various hepatobiliary disorders, were determinated for serum LN-LAP by the method of Goldbarg et al. Alteration of hydrolysis rate of LN-LAP by addition of D-leucine was investigated in 136 sera. Mobility of LN-LAP isozmes was measured by paper or starch block electrophoresis in 20 samples of serum, bile or hepatic tissue. LN-LAP activity was also determinated in 21 tissue specimens of the liver or stomach and 2 bile samples. The results are summarized as follows. 1) Patients with cancer of the liver demonstrated the highest level of serum LNLAP activity. Patients with malignant obstructive jaundice, cholangioltic hepatitis, cholelithiasis, viral hepatitis and cholecystitis showed the second place of higher values. Patients with malignant tumors other than liver, bile duct and head of pancreas usualy gave normal LN-LAP results. 2) Ninety eight percents of patients with cirrhosis of the liver showed below 300 units, while 91 percents of patients with primary cancer of the liver showed more than 300 units and 82 percents of patients with metastatic cancer of the liver showed over 300 units. Measurement of serum LN-LAP activity is usefulll in detection of occult cancer in the liver. 3) It was supposed that increased serum LN-LAP activity in patients with cancer of the liver is attributed to release of LN-LAP from injured hepatic cells, transference of LN-LAP from congested bile to circulating blood, increase of production of LN-LAP in the vicinity of tumor and increase of LN-LAP in residual normal hepatic tissue in the tumor bearing conditions. 4) It was difficult to differenciate the causes of biliary obstruction by the level of serum LN-LAP activity. 5) Serum LN-LAP activity was, unlike alkalin phosphatase, scarcely elevated in patients with bone or parathyroid diseases. 6) Five out of 60 patients with malignant tumor showed inhibition of serum LNLAP by addition of D-leucine, while 5 out of 43 normal subjects or patients with benign diseases other than hepatitis showed activation by D-leucine. 7) Electrophoretic mobility of serum LN-IAP isozymes showed one peak coincided with al-globulin in normal subjects and most of patients with hepatitis. The second peak coincided with α1-globulin or the third peak coincided withβ-globulin was demonstrated in patients with obstructive jaundice or liver cancer. LN-LAP in hepatic tissue demonstrated 3 peaks with a major one being coincided with al-globulin and LN-LAP in bile showed a major peak coincided with, β-globulin. 8) LN-LAP activity in hepatic tissue was diminished in patients with hepatitis and cirrhosis of the liver, while it was elevated in patients with cancer. LN-LAP activity in cancerous tissue per se showed an increase in cancer of the stomach and a decrease in cancer of the liver.
Amines produced from amino acid by enterobacteria are not sufficiently detoxicated in affected liver to manifest various infavourable effect. Amines are produced by amino acid decarboxylase of bacteria in digestive canal. In this investigation, tyrosine decarboxylase activity and tyramine production of various enterobacteria were especially studied. Tyrosine decarboxylase activity was determined by manometric method and tyramine was proved by paperchromatography or as the crystal. 1) Tyramine production from tyrosine was examined for 1004 strains of enterobacteria isolated from feces of 84 individuals including healthy persons and patients with some digestive diseases. Gram negative bacilli producing tyramine were proved only in a low percentage, among them, kiebsiella was highest in percentage, followed by proteus and pseudomonas, then by escherichia. However majority of gram positive cocci, including all of St. faecalis tested, produced tyramine. 2) There was no distinction between healthy persons and patients as to tyramine producing enterobacteria. It was thought that production of tyramineby enterobacteria would be enhanced in the case of coexistence of particular strain and tyrosine in high content in digestive canal. 3) Tyrosol, p-hydroxyphenyllactic acid, p-hydroxyphenylpropionic acid and p-hydroxyphenylacetic acid were proved as the decomposition products of tyrosine with both tyramine producing and non-producing strains. 4) Intensity of tyramine production by enterobacteria was estimated by the Warburg manometric method in adition to paper chromatographical identification and chemical isolation as crystal. Data of the three comparative investigation kept a considerble parallelism.