Mechanism of stimulation of DNA synthesis induced by epinephrine in primary culture of adult rat hepatocytes.

Addition of epinephrine to primary cultured adult rat hepatocytes stimulated their DNA synthesis dose-dependently, especially in presence of insulin and epidermal growth factor. This effect of epinephrine was strongly inhibited by an alpha 1-antagonist, prazosin, but not by a beta-antagonist, propranolol, and was also slightly inhibited by an alpha 2-antagonist, yohinbin. These results indicate that the stimulation of DNA synthesis of hepatocytes by epinephrine is mediated predominantly by an alpha 1-action. 12-o-Tetradecanoylphorbol-13-acetate (TPA) or Ca2+-ionophore A-23187 stimulated DNA synthesis of Swiss 3T3 cells, but did not induce DNA synthesis of hepatocytes either singly or in combination. The fact that pretreatment of hepatocytes with TPA caused down-regulation of the stimulatory effect of epinephrine on DNA synthesis of hepatocytes within 15 min suggested that the effect of epinephrine on hepatocytes is mediated by its alpha 1 receptor and that TPA activated protein kinase c in the hepatocytes. Addition of dibutyryl cGMP did not induce DNA synthesis of hepatocytes. Therefore, the alpha 1-action of epinephrine that induce stimulation of DNA synthesis of primary cultured adult rat hepatocytes was apparently not mediated by either activation of phospholipid-dependent protein kinase or Ca2+ mobilization. Possible alternative mechanism was discussed.

Purification and characterization of the 26S proteasome complex catalyzing ATP-dependent breakdown of ubiquitin-ligated proteins from rat liver.

An ATP/ubiquitin-dependent proteasome complex with an apparent sedimentation coefficient of 26S was purified from rat liver to near homogeneity by an improved method based on procedures reported previously. Two electrophoretically distinct forms of the 26S complex, named 26S alpha and 26S beta, with very similar subunit compositions were found not only in purified preparations but also in crude extracts, indicating that the 26S proteasome is present as two isoforms. The 26S proteasome was shown to degrade multi-ubiquitinated, but not unmodified, lysozymes in an ATP-dependent fashion, to have ATPase activity supplying energy for proteolysis, and to contain isopeptidase activity to generate free ubiquitin Mg2+/ATP-dependently. The 26S proteasome also catalyzed the ATP-independent hydrolyses of three types of fluorogenic peptides with basic, neutral, and acidic amino acids at their cleavage sites, respectively. These peptides are also good substrates for the 20S proteasome, but their degradation by the free 20S proteasome and by its assembled form in the 26S complex differ markedly, suggesting a functional difference between the two forms of proteasomes. Electrophoretic and immunochemical analyses showed that the large 26S complex was composed grossly of two different structures: a core 20S proteasome with multicatalytic proteinase functions and an associated part possibly with a regulatory role. These two structures both consisted of multiple polypeptides with molecular masses of 21-31 and 35-110 kDa, respectively. The subunit multiplicity of the rat 26S proteasome closely resembled that of the human counterpart, showing only minor species-specific differences in certain components. The assembly of this multi-component complex was found not to involve a sulfhydryl bond. Electrophoretic peptide mapping with lysyl-endopeptidase indicated the non-identity of the multiple subunits of the 26S proteasome. From these structural and functional characteristics, the 26S proteasome, which is widely distributed in mammals, is suggested to be a new type of multi-molecular complex catalyzing the soluble energy- and ubiquitin-dependent proteolytic pathway.

Bile-induced DNA strand breaks and biochemical analysis of bile acids in an experimental model of anomalous arrangement of the pancreaticobiliary ducts.

A canine experimental model for the anomalous arrangement of the pancreaticobiliary ducts (APBD) was made to investigate the effects of bile acids on carcinogenesis. Seven adult mongrel dogs underwent dorsal pancreatico-cholecystostomy to serve as a functional model for APBD, and six dogs underwent the same procedure with the pancreatic duct ligated as a control group. Bile from the gallbladder was taken 14 months after surgery for bile acid analysis by HPLC. DNA strand breaks in HeLa cells induced by the bile were also investigated in situ by nick translation method. As a result, the fraction of cholic acid tended to be lower, and that of deoxycholic acid slightly higher in APBD-dogs (N.S.). The ursodeoxycholic acid percentage in APBD-dogs significantly decreased compared with that in the control and normal dogs (p < 0.05). Extremely high frequency of DNA strand breaks was shown in only two out of seven APBD-dogs. In those two dogs, the cholic acid percentage decreased and that of deoxycholic acid increased extremely. These findings suggest that the alteration of the bile composition in APBD caused frequent DNA strand breaks and repair which might lead to gene mutation and biliary tract carcinoma.

Congenital arteriovenous fistula of the superior mesenteric vessels.

Arteriovenous (AV) fistula of the mesenteric vessel is a rare lesion and is most often iatrogenic or traumatic in origin. An infant boy with congenital superior mesenteric AV fistula is described. The present case is the first reported in a newborn.

An experimental study on gastroschisis using fetal surgery.

Gastroschisis models were successfully created in three groups of fetal rabbits by means of fetal surgery. Anatomical and pathologic assessments were carried out on the neonates that had experimental gastroschisis. In group I surgery was performed on the fetuses of 21 days gestational age; in group II on the fetuses of 23 days; and in group III on the fetuses of 26 days, respectively. Five cases of experimental gastroschisis were made in each group. The body weights of those with gastroschisis were low as compared with those of normal neonates. Intestinal length decreased and wall thickness increased as the result of 5--10 days evisceration. Abdominal cavity decreased in volume in all cases and could not accommodate the eviscerated organs. The number and density of distal mesenteric vessels (vasa recta) was high in the shortened intestine in gastroschisis models. Therefore it can be reasonably assumed that the intestine is only contracted and the possibility of return to normal length remains.

Does the type of anomalous arrangement of pancreaticobiliary ducts influence the surgery and prognosis of choledochal cyst?

A new classification of the anomalous arrangement of pancreaticobiliary ducts (APBD) has been proposed following amendments and modificiations of the previous one. Fifty-one cases of choledochal cyst complicated with APBD were extensively examined and analyzed for clear visualization of the APBD system to make a standard classification. APBD were classified broadly into three types: type I, type II, and type III with their subtypes. Type I of APBD was seen in 18 (35.3%), type II in 11 (21.6%) and type III in 22 (43.1%) cases, respectively. A radical operation, including complete excision of the dilated biliary duct followed by reconstruction of the biliary tract was the surgical treatment of choice, because the reflux of the pancreatic juice into the biliary tract is prevented. However, a dilated common channel or accessory pancreatic duct, according to the new Komi type Ib, IIb, and IIIc3 of APBD, could be the cause of relapsing pancreatitis leading to chronic pancreatitis, due to the formation of a protein plug or pancreatic calculus in the dilated duct, even after this radical operation. In two complicated cases of type IIIc3 of APBD in choledochal cyst, we performed a pylorus-preserving pancreatoduodenectomy as one of the operative methods of choice. Long-term follow-up, more than decades, is essential to evaluate the results of surgical procedure for choledochal cyst, especially in those complicated cases with type Ib, IIb, and IIIc3 of APBD according to the new Komi's classification.

A case of malignant melanoma of the transverse mesocolon.

A pediatric patient with malignant melanoma of the transverse mesocolon is reported. No cases of malignant melanoma occurring from the mesenterium have been previously reported in the literature.

Relation of patient age to premalignant alterations in choledochal cyst epithelium: histochemical and immunohistochemical studies.

Forty patients with choledochal cysts, 17 children and 23 adults, were treated by cyst excision in this institution (1972 to 1982). Histochemical and immunohistochemical studies were performed on tissue from the walls of these 40 cysts. The type and degree of histologic changes observed were found to correlate with patient age at time of cyst excision. Specimens from four patients less than two years of age at time of excision revealed epithelial desquamation, fibrosis, and minimal evidence of inflammation. Among specimens from 13 patients, 4 to 15 years of age, only two had an intact epithelial lining. Marked inflammation and intramural glandular structures were observed. Specimens from 23 patients, more than 15 years of age at the time of cyst excision, showed marked acute or chronic inflammation and destruction of the epithelial lining. In this age group, intramural glandular structures with goblet and argyrophil cells were regularly observed within the cyst walls, and gastrin and somatostatin-like immunoreactivity was detected in cells of these metaplastic glands. In association with these changes, in five of the 23 patients in the oldest age group, adenocarcinomas were observed in the cyst walls. These studies establish the increasing rate of epithelial metaplasia in the walls of choledochal cysts with advancing age, and suggest that these changes are a continuing process with malignancy one of the end results.

[Case of metastatic malignant melanoma responded to combination chemotherapy with DTIC].

A 60-year old woman with metastatic malignant melanoma who was well responded to a combination chemotherapy including DTIC was reported. She was noted a lentigo in the left first toe and histological examination revealed malignant melanoma in October 1978. Amputation of the left lower leg and dissection of the left inguinal lymph nodes had been done. OK-432 was injected as postoperative immunotherapy. She was readmitted to our hospital with the symptoms of pain and numbness of the left arm. Physical examination revealed a palpable mass in the left supraclavicular region. Incisional biopsy of the supraclavicular mass revealed metastatic malignant melanoma. She received a combination of 100mg DTIC i.v. for 5 days, 100 mg ACNU i.v. for one day and 1 mg VCR i.v. for one day (DAV chemotherapy) postoperatively. Subcutaneous injection of OK-432 with the dose of 5 KE per week was continued. Major side effects of DAV chemotherapy were nausea and transient leukocytopenia. No serious side effects were observed. On completion of the first course of DAV chemotherapy, abnormal shadow of the left apex was completely disappeared and on completion of the third course of DAV chemotherapy, high density area was markedly decreased in the cervical CT. She gained symptomatic reliefs and was discharged in August 1983. The combination chemotherapy including DTIC appeared to be effective in the treatment of metastatic malignant melanoma.

[Clinical trial of UFT in recurrent or advanced cancer].

UFT, antitumor agent containing uracil and futraful (FT-207), was given orally to 14 patients with recurrent or advanced carcinomas (6 gastric, 5 colorectal and 3 other carcinomas) and the clinical effects were evaluated. One PR (partial response) 1 NC (no change) and 9 PD(progressive disease) were obtained among 11 evaluable patients. Response rate was 9.1% (1/11), and overall response rate was 14.3% (2/14). According to Karnofsky's criteria, 2 responders (better than I-A) were obtained and response rate was 15.4% (2/13).

[Cooperative study of surgical adjuvant chemotherapy for colorectal cancer (third report): five-year results. Cooperative Study Group of Surgical Adjuvant Chemotherapy for Colorectal Cancer in Japan].

A randomized controlled study was carried out by the envelope method with 491 institutions in participation across the country in order to find an optimal surgical adjuvant chemotherapy for curatively resected colorectal cancer. The schedules for drug administration were different in four districts: ACNU + Futraful (FT) group and FT alone group in the Hokkaido-Shikoku district; the same schedule groups plus untreated group in the Chubu-Kinki district; MMC+FT group, FT alone group in the Tohoku-Kanto district; and ADM+FT group and FT alone group in the Chugoku-Kyushu district. The numbers of patients admitted to this study were 2,450 cases with colon cancer and 2,456 cases met the evaluation criteria of this study. The 5-year survival rate on the whole did not differ from combination therapy to single drug therapy in either colon cancer or rectal cancer, but in Dukes C rectal cancer the five-year survival rate tended to be higher with the combination therapies. In n2 (+) or a2(s) rectal cancer in particular, combination therapies with MMC and FT and with ADM and FT achieved significantly higher five-year survival rate, and the rate of local recurrence was significantly lower with ADM+FT.

[Cooperative study of surgical adjuvant chemotherapy of colorectal carcinoma (second study): a preliminary report. Cooperative Study Group of Surgical Adjuvant Chemotherapy of Colorectal Cancer in Japan].

In a prospectively randomized study, the effect of adjuvant chemotherapy with mitomycin C (MMC) and tegafur (FT) on survival and recurrence was analyzed in 2,477 evaluable patients with colon or rectal cancer who underwent macroscopically curative resection. Patients (1,256 with colon cancer, 1,221 with rectal cancer) were divided into the treatment group (group A) and the control group (surgical resection only, group B). In group A, chemotherapy consisted of intravenous administration of MMC (12 mg/m2 on operative day, followed by 6 mg/m2/2 months, 6 times) and oral administration of FT (800 mg/day for one year). No serious adverse effects were observed in the treatment group. There was no significant difference between group A and group B in three-year survival rate. The disease-free interval curve in group A of rectal cancer revealed significantly better results than group B (p = 0.044). There was no difference in the incidence of local recurrence at three years after operation between group A and group B. The incidence of metachronous liver metastases in group A of rectal cancer was significantly lower than in group B (p = 0.036).

[Cooperative study of surgical adjuvant chemotherapy of colorectal carcinoma (second study): 3-year survival after surgery of non-curatively resected patients. Cooperative Study Group of Surgical Adjuvant Chemotherapy of Colorectal Cancer in Japan].

A prospective randomized study was carried out to evaluate the efficacy of surgical adjuvant chemotherapy for non-curatively resected colorectal carcinoma patients as a collaborative study at 428 institutes in Japan from Jan., 1984 to Dec., 1985. A total of 1,138 patients were entered in this study. All patients were divided according to disease location (colon or rectum) and received one of two chemotherapeutic regimens after surgery (Regimen C: MMC 12 mg/m2 i.v. at operation day and 6 mg/m2 in every 2 months +UFT 600 mg/day p.o., D: the same dose of MMC + UFT 400 mg/day p.o.) randomly. 556 cases (colon carcinoma regimen C: 148, D: 185 cases, rectal carcinoma regimen C: 94, D: 129 cases) were evaluable at this presentation. Three-year-survival rate of histologically confirmed non-curatively resected colon carcinoma patients was higher in the group receiving regimen C than in regimen D. But no significant difference was found among the other groups.

[Cooperative study of surgical adjuvant chemotherapy of colorectal cancer (first report): Investigation of background factors and adverse effects. Cooperative Study Group of Surgical Adjuvant Chemotherapy of Colorectal Cancer in Japan].

In order to evaluate the efficacy of surgical adjuvant chemotherapy in patients undergoing gross curative resection for colorectal cancer (excluding m and sm cancer), a randomized controlled study was conducted from January, 1982 to October, 1983. Four hundred and ninety-one institutions participated in this study. The schedules for drug administration differed according to each district. In the Hokkaido and Shikoku districts, the patients were divided into the following two groups, one was a combination of ACNU and Futraful (FT) and the other was FT only. In the Chubu and Kinki districts, three groups were studied, namely those receiving a combination of ACNU and FT, those receiving FT only and those given no adjuvant chemotherapy. In the Tohoku and Kanto districts, a combination of MMC and FT and administration of FT only were studied. In the Chugoku and Kyushu districts, the patients were divided into a combination of ADM and FT, and FT only group. Among the 3,926 registered cases, 3,421 cases were valid for the study. As to the background factors, there were no significant differences among the groups in each district. There were no significant differences in one-year survival rates and one-year disease-free rates. No serious adverse effects were observed in any of the groups.

[Cooperative Study of Surgical Adjuvant Chemotherapy for Colorectal Cancer (third report): five-year results after surgery].

Beginning in January 1984, in collaboration with 357 institutions nationwide, comparative studies on curative resection cases of colorectal cancer and a control group were conducted for 2 years. The following method of administration was employed; Group A received 12 mg/m2 of MMC on the day of operation, followed by 6 mg/m2 every 2 months 6 times. From the 2nd week after the operation, 800 mg/body/day of Futraful was administered for one year. Group B received no treatment. Altogether 2786 cases were collected and, excluding ineligible cases, 2477 evaluable cases were analyzed. There was no difference in either survival rate or disease free rate, but after the bias of background factors was corrected by the hazard model, the disease free rate was better with Group A, and there was a significant difference with the rectum. According to the Dukes classification, in the Dukes C cases with the rectum Group A had higher rate of survival, disease free rate and lower rate of metastasis to the liver and local recurrence of cancer.

[Cooperative Study of Surgical Adjuvant Chemotherapy for Colorectal Cancer (fourth report): five-year results after surgery].

In collaboration with 428 institutions nation-wide, comparative studies on the optimal daily dosage of UFT were carried out for non-curative resection cases of colorectal cancer for two years from January 1984 to December 1985. The administration method employed was as follows. Group C received 12 mg/m2 of MMC on the day of operation, followed by 6 mg/m2 every 2 months 6 times, and from the 2nd week after the operation 600 mg/body of UFT was continuously administered every day for 1 year. Group D received 400 mg/body of UFT every day and the same amount of MMC as Group C. Analysis was made of 556 evaluable cases. There was no difference in the survival rate between the two groups. With respect to the dosage per body weight (kg), 8-12 mg/kg of UFT produced good prognoses and less side effects than the group which received more than 12 mg/kg. It seems necessary to administer UFT at its optimal dosage for colorectal cancers.

[Adjuvant chemotherapy for gastric cancer--the third study. First report: central randomization by telephone method and analysis of patients' background factors].

JFMTC conducted the third adjuvant chemotherapy study for gastric cancer patients after curative surgery from 1982 to 1983. Patients were randomly allocated to one of the three arms by telephone method controlled at the headquarters of the foundation. This method resulted in the marked reduction of ineligible cases (253 cases, 6.0% of 4,236 cases), compared with those in the first study (18.3%). Main reasons of ineligibility were the violation of entry criteria due to the misjudgment of noncancer, duplicate cancers and operative curability. Telephone method for random allocation seems to contribute to the improvement in the data quality of a clinical trial.

[A cooperative study of surgical adjuvant chemotherapy of colorectal cancer (second report): 3-year survival rate. Cooperative Study Group of Surgical Adjuvant Chemotherapy of Colorectal Cancer in Japan].

To evaluate the efficacy of surgical adjuvant chemotherapy for patients with curatively resected colorectal cancer (excluding m and sm cancer), a randomized controlled study was performed from January 1982 to October 1983. The schedules for drug administration were different in four districts, and four randomly assigned protocols were studied using tegafur (FT), ACNU, MMC and ADM. A total of 4,906 cases from 491 institutions were entered and 4,206 cases were varied for the study. There were no significant differences in 3-year survival rate in each district protocol. According to Dukes C for rectal cancer patients, ADM + FT group was higher than the FT only group in 3-year survival rate (p = 0.092) and had a significantly longer survival than FT only group in 3-year disease-free rate (p = 0.011). The rate of local recidivation in colon cancer resected curatively was higher in ACNU + FT group than in FT only group (p less than 0.05). A tendency for decreased liver metastasis was observed in FT group compared with the control group, and liver metastasis of ADM + FT group was lower than that of FT only group. No serious adverse effects were observed in any protocol.

[Attempted marrow rescue with cryopreserved circulating stem cells following high-dose chemotherapy].

Circulating hematopoietic stem cells were collected by three consecutive leukaphereses during post-chemotherapy expansion of the stem cell pool in a 3-year-old boy with advanced and therapy-resistant neuroblastoma. The cells were fractionated by discontinuous Percoll gradient centrifugation, frozen in a programmed freezer and then stored in liquid nitrogen. Following high-dose chemotherapy, the cells were thawed rapidly and re-infused into the patient. Early evidence of marrow recovery was first noted at day 13 and the times required to achieve a granulocyte count of greater than 0.5 x 10(9)/L and a platelet count of greater than 50 x 10(9)/L were 21 days and 30 days, respectively. This new marrow-rescue operation may have potential in cancer therapy as an alternative to bone marrow transplantation and further clinical investigation is warranted.

[Effect of synthetic protease inhibitor on the sphincter of Oddi function in dogs].

Fundamental study in dogs have shown that the synthetic protease inhibitor has pharmaceutical properties characterized by effect on motility of the sphincter of Oddi. Synthetic protease inhibitors, gabexate mesilate and nafamostat mesilate have effects on motility of the sphincter of Oddi in dogs. The motor effect of synthetic protease inhibitor on the sphincter of Oddi has been investigated by manometric evaluation. Immediately after intravenous administration of gabexate mesilate (5, 10, 50, and 100 mg/kg/h) the motility of the sphincter of Oddi was inhibited dose-dependently, on the other hand after intravenous administration of nafamostat mesilate (0.5, 1, and 5 mg/kg/h) motility of the sphincter of Oddi was accelerated. Therefore it was suggested that these results shown some considerable problems in clinical use.