Close association between high serum ALT and more rapid recurrence of hepatocellular carcinoma in hepatectomized patients with HCV-associated liver cirrhosis and hepatocellular carcinoma.

We investigated whether or not a high serum alanine aminotransferase (ALT) level is associated with a more rapid recurrence of hepatocellular carcinoma (HCC) in hepatectomized patients with hepatitis C virus (HCV)-associated liver cirrhosis (LC) (HCV-LC) and HCC. Thirty-three hepatectomized patients with HCV-LC and HCC of a single nodule who had no histologic evidence of portal or hepatic vein invasion and who had been followed up for more than 3 years were included in the study. They were subdivided into two groups according to their serum ALT levels, ALT being a well-known marker of inflammatory necrosis in the liver. Seventeen patients whose serum ALT levels showed several peaks or plateaus above 80 international units (IU) were designated as the high ALT group, and 16 patients whose serum ALT levels showed a sustained low level below 80 IU until the first recurrence were designated as the low ALT group, and the interval between hepatectomy and the first recurrence was observed. In the high ALT group, HCC recurred within 3 years in 70.6% of the patients. In contrast, it recurred in only 18.8% of the low ALT group within the same period (p < 0.05). There was a significant difference (p = 0.0201) between the two groups in the cumulative nonrecurrence rate. The mean interval in recurrent patients between hepatectomy and the first recurrence in the high ALT group (23.6 +/- 2.8 months; mean +/- SE) was significantly (p < 0.02) shorter than that in the low ALT group (49.3 +/- 9.7 months). The expected interval between hepatectomy and recurrence was as short as 2.8 +/- 0.5 years (mean +/- SE) in the high ALT group, compared with 5.8 +/- 0.7 years in the low ALT group (p < 0.05). These results showed that the recurrence of HCC was accelerated in the high ALT group, suggesting that suppression of the rise in ALT level after hepatectomy by treatment with anti-inflammatory drugs may prolong the interval until recurrence by about 2 years in hepatectomized patients with HCC and HCV-LC.

Mutagenic activity of surface soil and quantification of 1,3-, 1,6-, and 1,8-dinitropyrene isomers in soil in Japan.

To clarify the mutagenic potential of nonagricultural surface soil in Japan, 110 soil samples were collected from five geographically different areas between November 1996 and March 1997, and organic extracts of the soil samples were examined by the Ames/Salmonella assay. Most of the soil extracts showed mutagenicity toward both strains TA98 and TA100 in the presence and/or absence of a mammalian metabolic activation system (S9 mix), suggesting that surface soil is largely contaminated with environmental mutagens. Soil samples collected at Hekinan, Kobe, and Osaka were highly mutagenic toward both strains, and their potencies toward TA98 without S9 mix were extremely high, inducing more than 12 000 revertants per gram of soil. On the other hand, soil samples from Muroran showed strong mutagenicity toward TA100 with S9 mix. Furthermore, 1, 3-dinitropyrene (DNP), 1,6-DNP, and 1,8-DNP in soil samples collected at 10 sampling sites in three metropolitan areas were quantified by fluorometric detection of the corresponding diaminopyrene isomers using high-performance liquid chromatography (HPLC). Three DNP isomers were detected in all soil samples, and the amounts of 1,3-, 1,6-, and 1,8-DNP isomers in the soil samples were 12-3270, 14-5587, and 13-6809 pg/g, respectively. The gross amount of three DNP isomers in surface soil collected at Hekinan was more than 10 ng per gram of soil. The highest contribution ratios of DNP isomers to the mutagenicity of soil extracts were observed for the samples collected at Osaka, and the total of the contribution ratios of three DNP isomers was about 50%. These results suggest that surface soil is largely contaminated with mutagenic compounds and that DNP isomers are one class of major mutagenic and carcinogenic compounds contaminating surface soil.

Mutagenicity of dog urine determined by blue rayon extraction and the ultramicro forward-mutation method.

The mutagenicity of 15 household dog urine specimens were measured by the combination of blue rayon extraction and ultramicro forward-mutation method with Salmonella Typhimurium TM677 strain. A good dose-response relation was observed between the urine volume and mutation frequency. The minimum amount of urine required was 20 ml or less. The specific mutation frequency of urine greatly varied from one dog to another. The average specific mutation frequencies in the presence and absence of S9 mix were 28.7 +/- 51.5 (x 10(-4)) and 12.0 +/- 13.3 (x 10(-4)), respectively, and there was no significant difference between them. The mutation frequency markedly increased after the ingestion of broiled fish. Ten human urines specimens showed a similar level of specific mutation frequency to that of the dog urine specimens in both the presence and absence of S9 mix.

[The significance of surgical treatment for T4 lung cancer].

A total of 114 pT4 patients underwent pulmonary resection from January 1980 to December 1993 at our hospital. The overall five-year survival rate of the 114 cases was 17%. There was no significance between the five-year survival rate of squamous cell carcinoma and that of adenocarcinoma. Five-year survival rates of 17 patients with N0 disease, 29 patients with N1 disease, 51 patients with N2 disease, 1 patients with N3 disease, and 16 patients with NX disease were 40%, 21%, 21%, 0%, and 0%, respectively. Five-year survival rate of 89 cases with single factor of T4 was 24% and that of 25 cases with more than one factor was 4% (0.05 < P < 0.1). In the patients with only single factor of T4, five-year survival rates of 6 patients with left atrium invasion, 13 patients with major vessel invasion, 9 patients with trachea/carina invasion, and 18 patients with vertebral body, were 50%, 38%, 22%, and 0%, respectively. Furthermore, in 38 pT4 patients with involvement only to single adjacent structure, five-year survival rates were 47% in curative operation (n = 17) and 10% in non-curative operation (n = 21) (P < 0.05). We conclude that surgical treatment for T4 lung cancer should be performed to patients with single structure invasion which is expected to have a curative operation. N0 disease is more favorable than N1 or N2 disease.

Mutagenicity of alkylhydrazine oxalates in Salmonella typhimurium TA100 and TA102 demonstrated by modifying the growth conditions of the bacteria.

Alkylhydrazines are important carcinogens. However, they show generally only weak mutagenicity and the activities reported from different laboratories are contradictory. We have developed a sensitive method to detect the mutagenicity of alkylhydrazines. The method is based on a modified preculturing procedure in the Ames test, the emphasis in the modification being a change in the growth period of tester strains. The optimal growth periods were found to be 11 h in Salmonella typhimurium TA100 and 5 h in Salmonella typhimurium TA102. We tested the mutagenic activity of 12 alkylhydrazines; 1,2-dimethylhydrazine, 1,2-diethylhydrazine, 1,2-dipropylhydrazine, 1,2-dibutylhydrazine, 1,1-dimethylhydrazine, 1,1-diethylhydrazine, 1,1-dipropylhydrazine, 1,1-dibutylhydrazine, methylhydrazine, ethylhydrazine, propylhydrazine and butylhydrazine. All 12 alkylhydrazines were clearly mutagenic in Salmonella typhimurium TA102, and 10 hydrazines were mutagenic in Salmonella typhimurium TA100, both in the absence of S9 mix. The mutagenicity was inhibited by the addition of S9 mix or bovine serum albumin. This suggests deactivation of the mutagens by proteins.

Ventromedial hypothalamic lesions increase gastrointestinal DNA synthesis through vagus nerve in rats.

BACKGROUND: It was recently reported that ventromedial hypothalamic (VMH) lesions produced an increase in gastrointestinal DNA content in rats. In the present study, the mechanism of this alteration was examined. METHODS: The DNA content and synthesis after VMH lesioning in rat gastrointestinal tracts were determined. RESULTS: Total content of DNA in stomach and small intestine began to increase at 3 days and continued to increase for 7 days, whereas DNA content in the large intestine began to increase at 3 days and maintained the same level until 7 days after VMH lesioning. DNA synthesis of these organs increased and reached maximum at 3 days and then decreased to the initial level 7 days following the lesions. This increase in DNA content and synthesis in these organs was largely inhibited by bilateral subdiaphragmatic vagotomy or the administration of atropine, a cholinergic blocker, but not by the administration of anti-insulin antibody. CONCLUSIONS: VMH lesions induce cell proliferation in the rat gastrointestinal tract by the firing of vagus nerve activity mainly through the cholinergic receptor mechanism.

Hepatocellular carcinoma: treatment with percutaneous ethanol injection and transcatheter arterial embolization.

The therapeutic effectiveness of a new combination therapy--pretreatment with transcatheter arterial embolization (TAE) and subsequent percutaneous ethanol injection (PEI)--for solitary large (> 3.0 cm in diameter) primary hepatocellular carcinoma lesions was compared with that of TAE alone. With TAE alone, a partial response of the tumor was seen in only 10% of the patients, and the 1-, 2-, and 3-year survival rates were calculated to be 68%, 37%, and 0%, respectively. Histologic examination of specimens obtained at hepatectomy showed that TAE alone caused complete necrosis in only 20% of the tumors. In contrast, PEI combined with TAE significantly (P < .05) increased the partial response rate (45%) and significantly (P < .01) prolonged the 1-, 2-, and 3-year survival rates (100%, 85%, and 85%, respectively). Combination therapy caused complete histologic necrosis in 83% of the tumors. It also was significantly (P < .05) better than TAE alone in terms of rate of primary tumor recurrence during follow-up.

Role of vagus nerve in increased DNA synthesis after hypothalamic ventromedial lesions in rat liver.

We recently reported that ventromedial hypothalamic (VMH) lesions produced an increase in DNA content in rat liver. In the present study, we examined the mechanism of increased hepatic DNA content produced by VMH lesions. Hepatic DNA content began to increase 3 days after VMH lesioning and continued to increase until 7 days after the lesioning. Hepatic DNA synthesis increased and reached maximum 3 days after the lesioning and then decreased to the initial level 7 days after lesioning. The increased DNA content and synthesis after VMH lesioning were completely inhibited by vagotomy (hepatic vagotomy or bilateral subdiaphragmatic vagotomy) and largely inhibited by the administration of atropine but not by administration of anti-insulin antibody. These results suggest that vagus firing produced by VMH lesions stimulates DNA synthesis in rat liver mainly through cholinergic receptor mechanisms.

Collaborative study using the preincubation Salmonella typhimurium mutation assay for airborne particulate matter in Japan. A trial to minimize interlaboratory variation.

A collaborative study has been performed over a period of 3 years to develop a suitable method for monitoring the mutagenicity of airborne particulate matter. The study was organized with 8 laboratories and performed in the following steps: (1) selection of a suitable technique for each process involved in the mutagenicity monitoring, (2) developing a tentative protocol by combining systematically the selected techniques, (3) evaluation of the protocol by intra- and inter-laboratory studies, (4) modification of the protocol according to the evaluation, and (5) evaluation of the modified protocol by conducting an interlaboratory study. We found a suitable method for mutagenicity monitoring of particles in the atmosphere. Airborne particles were sampled with a high-volume sampler, the samples were stored at -80 degrees C, extracted by sonication using dichloromethane, solvent-exchanged, and assayed by the preincubation method using Salmonella typhimurium TA98 and TA100. The observed mutagenic activity was normalized with that of an internal standard. Round robin tests revealed that the method resulted in excellent reproducibility. The coefficient of variation for mutagenic activities of airborne particulate samples collected in various districts of Japan were in the range of 14.7 +/- 6.6% to 19.6 +/- 4.0% for strains TA98 and TA100 with and without metabolic activation. We also found that the plate incorporation method was equivalent to the preincubation method for airborne particulate extracts.

Results of a comparative study on the Salmonella pre-incubation and plate incorporation assays using test samples from the IPCS collaborative study.

Three complex mixtures (air particles, diesel particles and a coal tar fraction) and two pure compounds (benzo[a]pyrene and 1-nitropyrene) were tested in both the pre-incubation and the plate incorporation assay employing Salmonella typhimurium TA98 and TA100. Each experiment was conducted independently 2 or 4 times in duplicate in the presence and absence of metabolic activation. The mutagenic activities were calculated by least squares linear regression from the slope of the linear portion of each dose-response curve. Although slightly higher mutagenic activity was observed in the pre-incubation assay for the two pure compounds and with the plate incorporation assay for the diesel particulate sample, the overall data from both assays gave similar values and good correlations in TA100 and TA98. The results indicate that the pre-incubation assay could be used for these samples instead of the plate incorporation assay.

Detection of direct-acting mutagens in ambient air: a comparison of two highly sensitive mutagenicity assays.

Ambient air has been shown to contain numerous hazardous pollutants, many of which are known or suspected carcinogens and mutagens. Bioassays play a prominent role in the characterization of these genotoxic pollutants, and as new test methods are developed, it is incumbent upon researchers to evaluate assay performance and report relative merits. In this study, two Salmonella test methods (the spiral and preincubation assays) were assessed to determine their usefulness as screening methods for monitoring direct-acting mutagens in ambient air. The spiral assay automates the conventional plate-incorporation assay and has been shown to reduce the labor, materials, and sample mass required to perform mutagenicity testing. The preincubation assay has been shown to enhance test sensitivity for certain classes of compound, thereby reducing the amount of sample required for dose-response analysis. Both assays were used to test organic extracts of airborne particulate matter collected in Tokyo during the winters of 1988 and 1990. In addition to the conventional tester strains TA98 and TA100, two newly developed YG strains were evaluated. Strains YG1024 and YG1029-derived from TA98 and TA100, respectively-contain an acetyltransferase plasmid that confers upon the strains greater sensitivity towards nitroarenes. Results from this study indicated that both assays were able to detect direct-acting mutagens in the Tokyo air samples. The mutagenic activity associated with the samples was directly related to the particle mass present in a given volume of air. Mutagenic response was greater in the spiral assay relative to the preincubation assay, especially when YG tester strains were used. The YG strains were significantly more sensitive to mutation than the TA strains in both assays, which suggests that nitroaromatics are an important class of genotoxic contaminant present in Tokyo air.

Hepatocellular carcinoma: treatment with a combination therapy of transcatheter arterial embolization and percutaneous ethanol injection.

In an attempt to obtain complete tumor necrosis in large hepatocellular carcinoma (HCC) lesions, the authors studied the clinical and histologic findings of a new combination therapy, percutaneous ethanol injection (PEI) with transcatheter arterial embolization (TAE) (pretreatment with TAE and subsequent PEI) in 15 patients with a single, large (3.0-9.0 cm in diameter), encapsulated lesion of HCC. Two weeks after TAE, PEI was performed under ultrasound guidance. A total of four to 11 injections were administered at a rate of one injection twice a week. During the follow-up period (range, 7-23 months), all lesions were reduced in size and no evidence of HCC was present at contrast material-enhanced computed tomography or angiography in nine of 11 patients who did not subsequently undergo surgery. Six patients had a follow-up of 1 year or more, for a 1-year survival rate of 100%. Four patients subsequently underwent surgical resection; complete necrosis of the tumor was observed in all four. The authors conclude that a combination of PEI and TAE is an appropriate treatment for patients with large, encapsulated HCC lesions who are poor surgical risks.

[A case of paroxysmal cold hemoglobinuria (PCH) in the elderly found by severe anemia].

CASE: 69 year-old female. Two weeks before the admission, the patient had suffered from upper respiratory tract infection. Severe anemia was detected (Hb was 6.0 mg/dl) and she received a transfusion. Four days after the transfusion, she became icteric and was transferred to our hospital on Feb. 3, 1990 for evaluation of anemia. Laboratory data on admission are as follows; Hb 6.3 g/dl, reticulocytes 128%, total bilirubin 4.6 mg/dl, indirect bilirubin 3.4 mg/dl, urobilinogen in urine positive, WR negative, direct anti-globulin test positive, indirect of the test negative, and Donath-Landsteiner test positive (both direct and indirect). Her red blood cells were classified as P2-positive. From these findings, non-syphilitic PCH was diagnosed. She required bed rest and therapy to protect her from cold exposure. The hemoglobin recovered to 8.3 g/dl. The D-L test was positive for 9 months. This is a rare case of PCH because the patient fell ill in old age and PCH was found because of her severe anemia in spite of her unawareness of hemoglobinuria.

Comparison of DNA contents of visceral organs in rats with ventromedial hypothalamic lesions and fed a high fat diet.

Bilateral lesions of ventromedial hypothalamus are followed by a number of changes including vagal hyperactivity and hyperinsulinemia. To investigate if cell proliferation occurs in visceral organs in rats with ventromedial hypothalamic (VMH) lesions and fed a high fat diet, we determined DNA contents of visceral organs (liver, pancreas, small and large intestines, spleen, kidney and heart) 1 and 4 week after VMH lesions or start of high fat diet. In rats with VMH lesions, DNA contents increased significantly in liver, pancreas, and small and large intestines at 1 week, and maintained the same levels until the 4th week. DNA contents increased most in the pancreas, followed by small and large intestines, and liver. DNA content did not change in spleen, kidney, or heart. In rats fed a high fat diet, there was no increase in the DNA content of these organs, except in the small intestine at 4 weeks. The results suggest that VMH lesions produce excessive DNA synthesis in visceral organs, whereas a high fat diet does not. VMH lesions may induce cell proliferation in visceral organs through vagal hyperactivity and/or changes of humoral growth factors.

Effect of paromomycin sulfate on endotoxemia in patients with cirrhosis.

We evaluated, in a randomized double-blind trial, the efficacy of oral paromomycin sulfate administration in the prevention of endotoxemia in 24 cirrhotic patients with endotoxemia. Renal function was evaluated by glomerular-filtration rate and renal plasma flow at the beginning and at the end of the study period. After the administration of paromomycin sulfate, 2 g/day for 4 weeks, endotoxemia disappeared in 10 out of 13 (76.9%) cirrhotic patients with endotoxemia, whereas it became negative in only 3 of the 11 (27.3%) treated with placebo, the difference being significant (P less than 0.05). With regard to correlation of endotoxemia with renal impairment, endogenous creatinine clearance and p-aminohippurate clearance were significantly improved (P less than 0.02) in those patients whose endotoxemia disappeared on paromomycin sulfate administration. We did not find significant improvement, however, neither in liver function or in blood coagulation tests in the same patients. Paromomycin sulfate seems to be effective in the prevention of endotoxemia and the associated renal impairment in cirrhosis in man.

Relationship between endotoxaemia and protein concentration of ascites in cirrhotic patients.

Endotoxaemia was investigated by the Limulus assay in 42 cirrhotic patients with ascites and in 33 without ascites. The incidence of endotoxaemia in the former group (59.5%) was significantly (P less than 0.05) higher than in the latter (36.4%). Correlation between endotoxaemia and specific gravity and concentrations of total protein, albumin, and globulin in ascitic fluid was studied in the group with ascites. The specific gravity of ascites in 25 patients with endotoxaemia was significantly greater than that in 17 patients without endotoxaemia (P less than 0.01). The concentration of total protein in patients with endotoxaemia (13.95 +/- 7.18 milligram, mean +/- SD) was nearly twice as high (P less than 0.01) as in patients without endotoxaemia (7.49 +/- 3.60 milligram). The protein content of those who showed reactions greater or equal to 2(+) in the Limulus assay (16.78 +/- 7.14 milligram) was a significantly (P less than 0.05) higher than in those with 1(+) reaction (11.26 +/- 6.33 milligram). Moreover, the concentration of albumin in patients with endotoxaemia (7.68 +/- 4.60 milligram) was more than twice that of the patients without endotoxaemia (3.39 +/- 1.58 milligram, P less than 0.01). On the other hand, globulin concentration in patients with endotoxaemia was 1.6 times that of patients without endotoxaemia (P less than 0.01). Similar differences were noted between endotoxaemic and non-endotoxaemic patients in the ascites-to-serum ratio in protein, albumin, and globulin. These results suggest that in liver cirrhosis endotoxaemia may cause an increase in protein concentrations in ascitic fluid, and that it may be a precipitating factor in the formation of ascites.