Masaya Nishimura

Studies on the Metabolism of d-Limonene (p-Mentha-1,8-diene): I. The Absorption, Distribution and Excretion of d-Limonene in Rats

Abstract1. The absorption, distribution and excretion of d-limonene were investigated in rats using the 14C-labelled compound.2. The highest concentration of radioactivity in blood was obtained 2 h after oral administration of [14C]d-limonene and most occurred in the serum fraction. Radioactivity in the tissues reached maximum 1 or 2 h after administration. Radioactivity in liver, kidney and blood was higher than in other tissues, but was negligible 48 h after administration. An autoradiographic study confirmed these findings of tissue distribution.3. About 60% of administered radioactivity was recovered from urine, 5% from faeces and 2% from expired CO2 within 48 h. In bile duct cannulated rats, about 25% of the dose was excreted in bile within 24 h.

The use ofd-limonene preparation as a dissolving agent of gallstones

The dissolving mixture is administered through a choledochal drain to treat postoperatively retained cholesterol gallstones. It is preparted by mixing 97.0 parts of d-limonene with 2.1 parts of polysorbate 80 and 0.9 part of sorbitan monooleate, a mixture of which may easily reach the surface of the gallstones which are wetted by bile. The d-limonene preparation was found to be safe both in laboratory experiments and clinical trials. Before applying the preparation, the usual choledochal drain must be replaced with a recently developed catheter made from epichlorohydrine rubber, which is chemically resistant to the preparation. Three cases of retained gallstones are described where the preparation was successfully used. In the fourth case treatment with the preparation was tried in lieu of surgery but was not successful due to other complications. However, some dissolution of retained stones was observed. There were no postoperative complaints in the long-term follow-up of some cases for more than 2 yr after treatment with the preparation. This procedure promises to be of value because retained cholesterol stones may be dissolved without the necessity of further surgery.

Simultaneous occurrence of multiple gastric carcinomas and familial polyposis of the colon

Upper gastrointestinal survey in ten patients with familial polyposis of the colon revealed the presence of polyps and/or carcinomas of the stomach or duodenum in eight, including two cases of associated multiple gastric carcinomas with concomitant presence in one case of multiple gastroduodenal adenomatous polyps and in the other without presence of polyp. The rather frequent association of gastroduodenal polyps and cancerous lesions in familial polyposis suggests such association to be a part of the manifestation of this inherited disease and possible malignant change occurring in gastroduodenal mucosa as is generally supposed to be the case in large intestine.

Dissolution of human gallstone

A new water-soluble preparation of terpene derivative, natural limonene with polyoxyethylene hydrogenated castor oil was introduced. This preparation made it possible to dissolve completely 1.412 to 3.250 gm human gallstones, such as pure cholesterol stone, cholesterol pigment-calcium stone in extraordinary short time, about 2 hoursin vitro. And remarkable dissolving effects were shown on 1.925 to 2.801 gm human cholesterol stone placed into the pig gallbladder after only two times of instillation of this preparationin vivo. Histological examination after this instillation was within normal limits on various organ of pig, such as the gallbladder, common bile duct, duodenum, small and large intestines, liver, kidneys, suprarenal glands and sternal-marrow. LD50 value of this new solvent was higher than 5g/kg, therefore it was revealed as non-toxic preparation. These results indicate that this new preparation can be applied clinically with safety especially to cholesterol stone. Moreover dissolving effect was shown even on bile pigment-calcium stone and fatty acid-calcium stone, however the dissolving velocity was not so high as that of cholesterol stone. When this new preparation will be used clinically, the conventional surgical procedures on residual gallstone can be improved.

IMMUNOTHERAPY OF CANCER PATIENTS WITH BACILLUS CALMETTE‐GUÉRIN: SUMMARY OF FOUR YEARS OF EXPERIENCE IN JAPAN

Active immunotherapy with living BCG was conducted on 98 patients with various types of cancer. The candidates for this therapy were patients with residual or inoperable cancer of the colorectum, liver, breast, biliary tract, lung, and other organs with a follow-up of 4-58 months. Eleven of the 98 (11%) were able to survive for as long as 37-58 months (mean survival time 42.5 months) because of this treatment and are still living. Another 11 patients are also alive more than 24 months after starting treatment. Thirty-seven patients, however, succumbed within 12 months despite BCG immunotherapy. On the other hand, 37 patients in the control group, who shared the same clinical status and did not receive BCG therapy during this period, underwent unhappy courses for 2-12 months (mean survival time 8.7 months). The pretreatment immunoresponsiveness of these 98 patients was suppressed, as measured by the following immunologic parameters: T-cell subpopulation in the peripheral blood, stimulation index of PHA, and skin tests to DNCB, KLH, PPD, and PHA. All of these parameters improved shortly after initiation of BCG injections in 22 patients who survived more than 24 months. In contrast, in patients who died within 12 months, immunoresponsiveness remained suppressed throughout the course. This result has suggested that there was an apparent correlation between the effectiveness of BCG and immunoresponsiveness. In addition, a good correlation was observed between the duration of inflammatory reactions at BCG injection sites and clinical prognoses. Moreover, it was shown that a relatively high amount of BCG (20-80 mg as an initial dosage) and repeated injections of living BCG were necessary to obtain a sufficient enhancing effect on the immunocompetency of these late-stage cancer patients. The most conventional criterion used to determine an optimal time for booster injections of BCG was measurement of the PPD-evoked skin reaction at the BCG injection site, that is, Kochs phenomenon. When a marked flare-up reaction of more than 2.5 X 2.5 cm in size was observed, the effect of BCG was considered to be continuing, and no additional booster injection was needed. The mean interval between the first and second BCG injections was 6.2+/-1.1 months in patients who survived more than 2 years. In contrast, the duration of this reaction was only transient in ineffective cases. The most frequent side effects of this therapy were fever and malaise; these complications occurred in 62% of the cases. No severe side effects, such as dissemination, anaphylactic shock, or granulomatous hepatitis, have been experienced throughout this study, even in patients to whom a total dosage of more than 200 mg of living BCG were injected.

Diagnosis and treatment of solitary nonparasitic hepatic cyst

Total of 10 cases of nonparasitic cyst of the liver encountered during the past 20 years have been presented together with the 287 cases collected from the Japanese literature up to 1973 inclusive. The advent of the newer effective diagnostic procedures in recent years made it possible to establish preoperative diagnosis resulting in rapid increase of reported cases. Among them celiac and superior mesenteric angiography coupled with echotomography seem to be the most effective. Differentiation from hydatid cyst presents problems for which angiographic signs offer valuable clues. Total excision of the cyst has been advocated and accomplished successfully because of the relative ease of enucleation from the surrounding structures.

Early diagnosis of gallbladder malignancy

The treatment of gallbladder malignancy has been rather unrewarding because of the difficulty in establishing diagnosis in early stage. A review of 34 cases of malignant gallbladder neoplasm encountered during the past 17 years has been made in order to aide diagnosis by careful evaluation of the case history, presenting signs, laboratory findings and radiological studies. Tenderness and a mass in right upper quadrant and nonvisualization of the gallbladder by cholecystography seem to be the most frequent presenting signs for suspecting gallbladder malignancy. Recent advance of celiac angiography seems to hold promise in establishing preoperative diagnosis thus allowing initiation of early effective treatment.

Gallbladder bile peptides and their structure

Human gallbladder bile was separated by salting-in column chromatography using silica gel ammonium sulfate system. The second fraction containing peptides was found to vary in cholelithiasis in relation to Fraction 1.To determine the structure of the peptides, bovine gallbladder bile was fractionated by salting out method with ammonium sulfate. Four peptides were isolated and their amino acid sequences were determined to be:1)β-Ala-(Trp1, His1, Ileu1, Phe1)2)3)Tyr-Gly4)

Vagotomy and Cholelithiasis

There has been a considerable divergence of opinion as to whether or not the incidence of gallstones increases after total vagotomy. As this surgical procedure is becoming more and more popular, a number of clinical studies have accumulated and these show there is a definite increase in the incidence of cholelithiasis after total vagotomy [5]. On the other hand, Fletcher and Clark failed to substantiate the claim. This controversy in clinical studies on the possible increase in incidence of cholelithiasis after total vagotomy led some investigators to turn to experimental studies. Here again, they seemed to encounter divergence. Thus, it is important to determine whether or not total vagotomy influences the composition of bile in such a way as to make it potentially more lithogenic.

Regurgitation cholelithiasis-with special reference to formation of fatty acid calcium stone

Three cases with fatty acid calcium stone were studied and discussed; two cases with the opening of pancreatic duct into a dilated cyst at the distal end of common bile duct, and one case with the triple-barreled gun shaped common channel of an anomalous remnant cystic duct, common bile duct and pancreatic duct into duodenum. The regurgitation of pancreatic juice was easily presumed from these anatomical findings. And these stones in the cyst were shown fatty acid calcium stone by gas liquid chromatographical analyses. This fact seemly supports our regurgitation cholelithiasis theory on formation of fatty acid calcium stone; this regurgitation played an important role in this stone formation.SummaryThree cases with fatty acid calcium stone were studied and discussed; two cases with the opening of pancreatic duct into a dilated cyst at the distal end of common bile duct, and one case with the triple-barreled gun shaped common channel of an anomalous remnant cystic duct, common bile duct and pancreatic duct into duodenum. The regurgitation of pancreatic juice was easily presumed from these anatomical findings. And these stones in the cyst were shown fatty acid calcium stone by gas liquid chromatographical analyses.This fact seemly supports our regurgitation cholelithiasis theory on formation of fatty acid calcium stone; this regurgitation played an important role in this stone formation.