The gastric sediment induced with the tetracycline fluorescence was studied in 25 out of 34 patients with cancer of the stomach. The result was positive in 74%(25/34) and negative in 26%(9/34). The negative cases include 6 cases which contained blood in gastric juice. Only 2 out of 34 patients with benign lesion of the stomach showed false positive. Those were suspected to be malignant lesion by the gross findings of the specimen at the surgical operation. but later histologically proved to be ulcus callosum. 7 strains of experimental ascitic tumors (Takeda, Yoshida, gDT-2) and tissue culture cells (Hela, G1I, L, FL, ) were also studied. In every case, fluorescence proved in the cytoplasm of these cells in the same manner, and no difference was observed among them. The slices of the subcutanouses implanted solid farm of Takeda sarcoma, which had been previously treated with tetracycline, were incubated with E D T A-2Na and Sod. Citrat, then the deminished intensity of fluorescence was observed. Therefore “Ca ions” might be one of the important factors in the induction of fluorescence. These fluorescence material was considered to be the same thing seen in the early stage in the other organs after the tetracycline administration. So it might be conculuded that in some tissues like cancer or bone those materials were kept longer than in the other tissue, in some unknown mechanisms. This method is simple and useful for the detection of cancer of the stomach.
A rapid preparation for gaschromatographic analysis of non-esterified fatty acids (NEFA) was investigated. Plasma lipid was extracted according to Shafrir's method in order to minimize the contamination of phospholipids and plasma NEFA was separated from the lipid extract by slightly modified Dole's method, in which 0.02% KOH in 90% ethanol was replaced by 0.02% KOH in 80% ethanol. The plasma NEFA was methylated with use of methanol-BF3and analysed by gaschromatography. Optimal condition for methylation of NEFA by ethanol-BF3was examined. The highest yield was attained by refluxing NEFA with 5 ml of 0.1% methanolic BF3for 3 min in a boiling water bath. Recovery of methylation of palmitic acid by methanol-BF3was 93% and was comparable with that obtained by the diazomethane method. The procedure for plasma NEFA extraction mentioned above demonstrated satisfactory recovery of palmitic acid, palmitoleic acid, stearic acid and margaric acid (90-92%) but lower recovery of oleic acid (89.2%) and linoleic acid (86.4%). The results obtained by the preparation showed no significant difference from those by the silicic column method of McCarthy. From these evidences, it has been concluded that the rapid preparation for gaschromatographic analysis of plasma NEFA is a reliable method for routine works in clinical chemistry.
The total and individual plasma non-esterified fatty acids (NEFA) of patients with liver diseases were analysed to elucidate the feature of abnormality of lipid metabolism in liver diseases, classified as acute hepatitis, liver cirrhosis and obstructive jaundice groups. The plasma level of total NEFA of fasting normal subjects averaged 436.5±31.0 uEp/l. In patients with liver diseases it was significantly higher: 576.6±60.0uEq/l for acute hepatitis group, 647.5±39.7uEq/l for liver cirrhosis group, 751.2±108.0uEq/l for obstructive jaundice group. Gaschromatographic analysis plasma NEFA revealed a significant increase of concentration of palmitic acid and oleic acid, especially in the groups of liver cirrbosis' and obstructive jaundice, but no significant change in concentration of linoleic acid was detected. The level of plasma NEFA correlated with icteric indexes, but not with the Zn T T values and alkaline phosphatase activities. The oral administration of glucose (50 g) in normal subjects lowered plasma NEFA to the minimum value at 60 min after which was restored to original level at 180 min after administration. But there was no significant difference in the pattern of fluctuation following the glucose intake between the control and liver diseases groups. NEFA mobilization from adipose tissue resumed from the glucose administration test is nearly within normal limits in the patients with liver diseases and the increase in plasma NEFA in the patients with liver diseases may be ascribed to decreased hepatic uptake of plasma NEFA. The current conclusion was supported by the report of Maezawa et. al.: the oral administration of ethanol lowered plasma NEFA level in normal subjects but not significantly in the patients with liver cirrhosis and the challenge with intravenous administration of ethanol in dogs lowered plasma NEFA level in artery and hepatic vein, but not significantly in CCl4-poisoning dog.
Some etiological studies of gall stones and of cholecystitis have ever been made and discussed from various angles and now it is apparent that the infection of the bile ducts plays important roles etiologically for them, aside from the problem whether it is concerned with them primarily or secondarily. But the infection theory contains some contradiction which is pointed out by various researchers in the fact that even many ba-cteria instillated into the gall bladder do not cause the formation of gall stones or cholelithiasis so easily. On the other, Prof. Matsukura has ever noticed the fact that in cholelithiasis and cholecystitis the concentration of bile acids, particularly of dihydroxycholanic acid, is conspicuously lowered in comparison with that of normal cases and advocated the significance of the low concentration of dihydroxycholanic acid in the infection of the bile ducts, based upon various clinical and experimental studies made in his laboratory. The author made some serial clinical and experimental studies on bile acids in bile, particularly the significance of the substance “Dihydroxycholanic Acid” in the infection of bile ducts, as one part of the etiological studies of cholecystitis and gall stones undertaken in the laboratory and obtained some results as follows. 1) The concentration of dihydroxycholanic acid in bile of healthy person or of patients with gastroduodenal ulcers is high, ranging from 30 to 60 mg/cc, while in cholelithiasis and cholecystitis, the levels are mostly in a range less than 20 mg/cc. 2) No bacteria were detected in the bile of healthy person or of the patients with gastroduodenal ulcer. But in bile of cholelithiasis or of cholecystitis, showing the level of dihydroxycholanic acid in a range less than 20 mg/cc, bacteria were found and isolated. 3) Coli-bacilli occupied the majority among the microorganisms isolated from the bile of cholelithiasis and of cholecystitis. According to these clinical and experimental studies, it can be presumed that in cholecystitis and in cholelithiasis, the colibacilli played the cardinal role and the infection of the bile ducts with coli-bacilli is closely related with the lowering of the concentration of dihydroxycholanic acid in bile. The author made a experimental study regarding the relation between the lowering of the concentration of the acid and the infection of the bile ducts in rabbit and following results were obtained. 4) The level of the dihydroxycholanic acid in the normal rabbit ranged from 70 to 110 mg/cc but if the occlusion of the bile ducts was artificially made at the Vater's ampullae where the choledochus joins the duodenun, the level increased slightly on the 2nd postoperative day but then decreased gradually up to the levels of 12 to 32 mg/cc on the 8th day, showing apparently the lowering tendency. 5) In normal rabbits which accepted colibacilli through the portal vein and in the group of rabbits of which liver function was damaged with carbon tetrachloride and 2, 4-diaminotoluene, coli-bacilli were detected in both of the groups but the numbers of the bacilli were markedly small and the slight increase of the bacilli was observed only in one case out of those with damage of the liver function. But in all of ten cases with the occlusion of the bile ducts, a great number of bacilli were observed. Moreover, in 8 cases, gall stones were constituted and in 2 cases bile sands were seen. 6) 1. In liver of normal rabbits which were instillated coli-bacilli through the portal vein, very slight inflammatory changes were seen, but at the gall bladder no inflammatory changes were seen grossly as well as histologically. 2. In livers of rabbits as well as in the gall bladder, with the damaged liver function and instillation of coli-bacilli through the portal vein, almost no inflammatory change was observed in the gall bladder.