Characterization of raft microdomains in bovine mammary tissue during lactation: How they are modulated by fatty acid treatments.

The objective of the current study was first to characterize lipid raft microdomains isolated as detergent-resistant membranes (DRM) from mammary gland tissue, and second to determine how dietary fatty acids (FA) such as conjugated linoleic acid (CLA), 19:1 cyclo, and long-chain n-3 polyunsaturated FA affect lipid raft markers of mammary cells, and to finally establish relationships between these markers and lactation performance in dairy cows. Eight Holstein cows were used in a replicated 4 × 4 Latin square design with periods of 28 d. For the first 14 d, cows received daily an abomasal infusion of (1) 406 g of a saturated FA supplement (112 g of 16:0 + 230 g of 18:0) used as a control; (2) 36 g of a CLA supplement (13.9 g of trans-10,cis-12 18:2) + 370 g of saturated FA; (3) 7 g of Sterculia fetida oil (3.1 g of 19:1 cyclo, STO) + 399 g of saturated FA; or (4) 406 g of fish oil (55.2 g of cis-5,cis-8,cis-11,cis-14,cis-17 20:5 + 59.3 g of cis-4,cis-7,cis-10,cis-13,cis-16,cis-19 22:6, FO). Mammary biopsies were harvested on d 14 of each infusion period and were followed by a 14-d washout interval. Cholera toxin subunit B, which specifically binds to ganglioside M-1 (GM-1), a lipid raft marker, was used to assess its distribution in DRM. Infusions of CLA, STO, and FO were individually compared with the control, and significance was declared at P ≤ 0.05. Milk fat yield was decreased with CLA and FO, but was not affected by STO. Milk lactose yield was decreased with CLA and STO, but was not affected by FO. Mammary tissue shows a strong GM-1-signal enrichment in isolated DRM from mammary gland tissue. Caveolin (CAV) and flotillin (FLOT) are 2 proteins considered as lipid raft markers and they are present in DRM from mammary gland tissue. Distributions of GM-1, CAV-1, and FLOT-1 showed an effect of treatments determined by their subcellular distributions in sucrose gradient fractions. Regardless of treatments, data showed positive relationships between the yield of milk fat, protein, and lactose, and the abundance GM-1 in DRM fraction. Milk protein yield was positively correlated with relative proportion of FLOT-1 in the soluble fraction, whereas lactose yield was positively correlated with relative proportion of CAV-1 in the DRM fractions. Infusion of CLA decreased mRNA abundance of CAV-1, FLOT-1, and FLOT-2. Regardless of treatments, a positive relationship was observed between fat yield and mRNA abundance of FLOT-2. In conclusion, although limited to a few markers, results of the current experiment raised potential links between variation in specific biologically active component of raft microdomains in bovine mammary gland and lactation performances in dairy cows.

Effects of abomasal infusion of conjugated linoleic acids, Sterculia foetida oil, and fish oil on production performance and the extent of fatty acid Δ9-desaturation in dairy cows.

The purpose of this study was to determine the effects of conjugated linoleic acid (CLA), Sterculia foetida oil (STO), and fish oil (FO) on milk yield and composition, milk FA profile, Δ(9)-desaturation activity, and mammary expression of 2 isoforms of stearoyl-coenzyme A desaturase (SCD-1 and SCD-5) in lactating dairy cows. Eight multiparous Holstein cows (69 ± 13 d postpartum) were used in a double 4 × 4 Latin square design with 28-d periods. For the first 14 d of each period, cows received an abomasal infusion of (1) 406 g of a saturated fatty acid (SFA) supplement (112 g of 16:0 + 230 g of 18:0) used as a control (CTL), (2) 36 g of a CLA supplement (13.9 g of trans-10,cis-12 18:2) + 370 g of SFA, (3) 7 g of STO (3.1g of 19:1 cyclo) + 399 g of SFA, or (4) 406 g of FO (55.2 g of cis-5,-8,-11,-14,-17 20:5 + 59.3 g of cis-4,-7,-10,-13,-16,-19 22:6). Infusions were followed by a 14-d washout interval. Compared with CTL, STO decreased milk yield from 38.0 to 33.0 kg/d, and increased milk fat concentration from 3.79 to 4.45%. Milk fat concentration was also decreased by CLA (2.23%) and FO (3.34%). Milk fat yield was not affected by STO (1,475 g/d) compared with CTL (1,431 g/d), but was decreased by CLA (774 g/d) and FO (1,186 g/d). Desaturase indices for 10:0, 12:0, and 20:0 were decreased, whereas the extent of desaturation of 14:0, 16:0, 17:0, and 18:0 was not affected by CLA treatment compared with CTL. Infusion of STO significantly decreased all calculated desaturase indices compared with CTL; the 14:0 index was reduced by 80.7%. Infusion of FO decreased the desaturase indices for 10:0, 14:0, 20:0, trans-11 18:1, and 18:0. The effect of FO on the 14:0 index indicates a decrease in apparent Δ(9)-desaturase activity of 30.2%. Compared with CTL, mammary mRNA abundance of SCD-1 was increased by STO (+30%) and decreased by CLA (-24%), whereas FO had no effect. No effect was observed on mRNA abundance of SCD-5. In conclusion, abomasal infusion of CLA, STO, and FO were shown to exhibit varying and distinct effects on desaturase indices, an indicator of apparent SCD activity, and mammary mRNA abundance of SCD-1.

The effect of oral appliances that advanced the mandible forward and limited mouth opening in patients with obstructive sleep apnea: a systematic review and meta-analysis of randomised controlled trials.

Oral appliances (OAs) have demonstrated efficacy in treating obstructive sleep apnea (OSA), but many different OA devices are available. The Japanese Academy of Dental Sleep Medicine supported the use of OAs that advanced the mandible forward and limited mouth opening and suggested an evaluation of their effects in comparison with untreated or CPAP. A systematic search was undertaken in 16 April 2012. The outcome measures of interest were as follows: Apnea Hypopnea Index (AHI), lowest SpO2 , arousal index, Epworth Sleepiness Scale (ESS), the SF-36 Health Survey. We performed this meta-analysis using the Grading of Recommendations, Assessment, Development and Evaluation (GRADE) system. Five studies remained eligible after applying the exclusion criteria. Comparing OA and control appliance, OA significantly reduced the weighted mean difference (WMD) in both AHI and the arousal index (favouring OA, AHI: -7.05 events h(-1) ; 95% CI, -12.07 to -2.03; P = 0.006, arousal index: -6.95 events h(-1) ; 95% CI, -11.75 to -2.15; P = 0.005). OAs were significantly less effective at reducing the WMD in AHI and improving lowest SpO2 and SF-36 than CPAP, (favouring OA, AHI: 6.11 events h(-1) ; 95% CI, 3.24 to 8.98; P = 0.0001, lowest SpO2 : -2.52%; 95% CI, -4.81 to -0.23; P = 0.03, SF-36: -1.80; 95% CI, -3.17 to -042; P = 0.01). Apnea Hypopnea Index and arousal index were significantly improved by OA relative to the untreated disease. Apnea Hypopnea Index, lowest SpO2 and SF-36 were significantly better with CPAP than with OA. The results of this study suggested that OAs improve OSA compared with untreated. CPAP appears to be more effective in improving OSA than OAs.

A collaborative study for the evaluation of lectin-reactive alpha-fetoproteins in early detection of hepatocellular carcinoma.

Lectin-affinity electrophoretic separation of serum alpha-fetoprotein (AFP) was carried out using AFP Differentiation Kits, which used Lens culinaris agglutinin-A (Kit L) and erythroagglutinating phytohemagglutinin (Kit P). Separated AFP bands were detected with a sensitive antibody-affinity blotting technique and determined quantitatively by densitometry, and the results were expressed as percentages of the intensity of total AFP bands. Sera from 424 patients with acute hepatitis, chronic hepatitis, liver cirrhosis, hepatocellular carcinoma, and extrahepatic tumors were assayed for proportion of AFP present as Lens culinaris agglutinin-A-reactive AFP (AFP-L3) and erythroagglutinating phytohemagglutinin-reactive AFPs (AFP-P4+P5). From the maximum Youden indices determined, cutoff levels were set at 15% for both AFP-L3 and AFP-P4+P5 to discriminate between patients with chronic hepatitis and liver cirrhosis and patients with hepatocellular carcinoma. AFP-L3 and AFP-P4+P5 showed sensitivities of 55.3 and 61.0% at specificities of 93.9% and 82.3%, respectively. Thirty-eight % of tumors that measured less than 20 mm in diameter were positive for AFP-L3 and AFP-P4+P5. AFP-L3 exceeded the cutoff level of 15% 4.0 +/- 4.9 months before detection of hepatocellular carcinomas by imaging techniques with a sensitivity of 48% and a specificity of 81%. Thus, these tests are useful for the early detection of hepatocellular carcinomas in patients with hepatitis or liver cirrhosis.

Immunohistochemical study of alpha-fetoprotein in rat embryos during ontogenesis.

In order to clarify the dynamics of alpha-fetoprotein (AFP) during ontogenesis, the appearance and localization of AFP and albumin in rat embryos were immunohistochemically investigated. AFP began to appear in the embryos on day 9 in some of the cells constituting the embryonic ectoderm. From day 10, both AFP and albumin became strongly stained in the yolk sac. The formation of liver anlage started on day 11; here AFP alone was clearly apparent in the primitive hepatocytes and also in some other cells, such as somites, or those of the central nervous system. After further development of the embryos, AFP, albumin, and transferrin were detected by means of serial section examination in the same hepatocytes. Besides being found in the liver, AFP began to be stained in the dorsal pancreas and mesonephric vesicles on day 12, in the stomach on day 13, and in the lung anlage and metanephros on day 14. Albumin was also localized in the same sites in these tissues, but the intensity of albumin staining was not as strong as that of AFP. Therefore, we assume that AFP plays an important role in the rapid growth of rats during the early gestational stages.

Antifibrotic effect of the herbal medicine Saiko-keishi-to (TJ-10) on chronic pancreatitis in the WBN/Kob rat.

Chronic pancreatitis is characterized by fibrosis. We reported an anti-inflammatory effect of the herbal medicine Saiko-keishi-to (TJ-10) on chronic pancreatitis. This study aimed to elucidate the antifibrotic effect of TJ-10. Four-week-old male WBN/Kob rats were fed a special pellet diet (MB-3) with or without TJ-10 (80 mg/100 g body weight) for 20 weeks. Pancreata were histopathologically examined at every 4 weeks, and the expression of fibrosis-related factors such as transforming growth factor beta1 (TGF-beta1), fibronectin (FN), alpha-smooth muscle actin (alpha-SMA), and type III collagen was analyzed. In untreated WBN/Kob rats, chronic pancreatitis developed at 12 weeks and progressed with marked fibrosis at 16 weeks, and the expression of TGF-beta1 and FN peaked at 12 weeks. However, in the TJ-10-treated rats, the rate of pancreatic fibrosis and the expression of TGF-beta1, FN, alpha-SMA, and type III collagen at 12 and 16 weeks decreased significantly compared to those in the untreated rats. These results suggest that TJ-10 inhibits the pancreatic fibrosis by the suppression of TGF-beta1 expression.

Expression of pancreatitis-associated protein (PAP) in rat spontaneous chronic pancreatitis: effect of herbal medicine Saiko-keishi-to (TJ-10).

Pancreatitis-associated protein (PAP) is almost absent in the normal pancreas but is overexpressed in acute pancreatitis. However, its expression in chronic pancreatitis (CP) is unknown. An herbal medicine Saiko-keishi-to (TJ-10) has long been used clinically for CP, but there is no experimental evidence of the effect of TJ-10 on CP. The aim of this study was to analyze the expression of PAP and the effect of TJ-10 in a spontaneous chronic pancreatitis model. Four-week-old male WBN/Kob rats were fed with a special pellet diet (MB-3), and TJ-10 (80 mg/100 g body weight/day) was orally administered for 16 weeks. The rats were killed at every 4 weeks, and pancreata were histopathologically examined. PAP messenger RNA (mRNA) in the pancreas was detected with a reverse transcription/polymerase chain reaction (RT-PCR) method. The cellular localization of PAP mRNA and protein was analyzed with in situ hybridization (ISH) and immunohistochemistry (IHC). PAP mRNA was expressed from 8 weeks, when the pancreas was still pathologically normal, and reached its peak at 12 weeks, when the pancreatitis first appeared. Then the expression of PAP mRNA was decreased gradually. TJ-10 suppressed the expression of PAP mRNA completely at 8 and 12 weeks. PAP mRNA was slightly expressed at 16 and 20 weeks. ISH and IHC confirmed the PAP mRNA and protein expression in the cytoplasm of acinar cells. These results suggest that PAP mRNA appears before CP, and its peak coincides with the onset of CP. TJ-10 suppressed the PAP expression and delayed the development of CP in the WBN/Kob rat.

Establishment of assay kits for the determination of microheterogeneities of alpha-fetoprotein using lectin-affinity electrophoresis.

Diagnostic kits for determination of alpha-fetoprotein (AFP) carbohydrate chain microheterogeneity were developed using lectin affinity electrophoresis with Lens culinaris agglutinin-A (LCA-A) and erythro-agglutinating phytohemagglutinin-E4 (PHA-E4). Separated AFP bands by electrophoresis were detected with high sensitivity by antibody-affinity blotting and immunoenzymatic amplification. Densitometry was used to apportion lectin reactive AFPs. The within-run S.D. for proportions of AFP bands was below 3%. Band intensity was linearly related to AFP concentration between 2 and 200 ng/ml. Profiles of lectin reactive AFPs were compared in serum samples from 55 patients having liver diseases. The average values of lectin reactive AFPs for chronic hepatitis and liver cirrhosis patients were both below 13%, but those of hepatocellular carcinoma patients were above 25%. Correlation of data with disease states suggests that the methods can greatly facilitate the discrimination between benign and malignant liver diseases.

Synergistic effect of oral UFT in combination with cisplatin on DNA synthesis in rat hepatocarcinomas.

We investigated the synergistic efficacy of CDDP and UFT, an oral anticancer derivative of 5-fluorouracil, on DNA synthesis of 3'-MeDAB-induced hepatocarcinomas in rats. Chronic treatment with CDDP slightly reduced tissue activities of thymidylate synthetase and thymidine kinase in the hepatocarcinomas, and, moreover, simultaneous administration of a low dose of UFT with CDDP markedly suppressed both enzyme activities, i.e., UFT and CDDP showed a potent synergism on DNA synthesis in hepatocarcinomas.

Inhibition by 1-(2-tetrahydrofuryl)-5-fluorouracil in combination with uracil of hepatocarcinogenesis induced by 3'-methyl-4-dimethylaminoazobenzene in rats.

It is known that a high incidence of hepatocellular carcinoma in rat liver can be induced by 3'-methyl-4-dimethylaminoazobenzene (3'-MeDAB). The administration of 3'-MeDAB in combination with 1-(2-tetrahydrofuryl)-5-fluorouracil and uracil (UFT) delayed the appearance of oval cells and the formation of hyperplastic nodules, which were observed in the liver from 3 and 5 weeks, respectively, after the onset of 3'-MeDAB feeding, and also delayed the transient increase of serum alpha-fetoprotein level, which transiently peaked at 5 weeks, and completely suppressed the transient increase of tissue thymidylate synthetase activity, but not thymidine kinase, which were induced by 3'-MeDAB at 5 weeks, and finally reduced markedly the incidence of hepatocarcinomas. These results indicate that the suppression of de novo synthesis in pyrimidine metabolism prevents hepatocarcinogenesis.

Detection of Altered Glycosylation of α-Fetoprotein Using Lectin-Affinity Electrophoresis.

α-Fetoprotein (AFP) is an oncofetal glycoprotein with a molecular mass of 70,000 Dalton and a carbohydrate content of approx 4%, i.e., one carbohydrate chain per molecule. High concentrations of AFP are found in the fetal serum as well as in amniotic fluid. Serum concentration of AFP in cord blood at birth range from 10,000-100,000 ng/mL, and it becomes suppressed to levels below 10 ng/mL by 300 d after birth (1). AFP is re-expressed frequently in hepatocellular carcinomas and yolk sac tumors, and much less frequently in gastrointestinal carcinomas.

Lectin affinity electrophoresis of alpha-fetoprotein detected by immunoenzymatic and chemiluminescent amplification followed by direct scanning.

Microdetermination of alpha-fetoprotein (AFP) glycoforms by lectin affinity electrophoresis followed by chemiluminescence reaction using horseradish peroxidase (POD) or alkaline phosphatase (ALP) in antibody-affinity blotting was developed. The intensity of chemiluminescence obtained by ALP was greater than that by POD; however, the coefficient of variation with POD was less than that with ALP. The optimized sensitivity of the chemiluminescence method with POD was two times that of the most sensitive colorimetric method currently available in terms of the chemiluminescence intensity per unit AFP concentration. The lower detection limit by the chemiluminescence method with POD (0.5 ng/ml) was much lower than that by the colorimetric method (3 ng/ml). Both methods gave identical percentages of lentil lectin- and erythroagglutinating phytohemagglutinin-reactive minor bands using a serum with 52 ng/ml AFP. This result indicates that microdetermination of AFP glycoforms by chemiluminescence after lectin-affinity electrophoresis was more sensitive than currently available methods and that it is potentially useful for clinical application.

Electrophoresis and electro-affinity transfer with specific antibodies to alpha-fetoprotein for detection of circulating immune complexes of alpha-fetoprotein.

A combination of agarose gel electrophoresis and a newly developed technique of electro-affinity transfer was applied to the detection of circulating immune complexes of human alpha-fetoprotein (AFP) and anti-AFP. After electrophoretic transfer to nitrocellulose membrane, to which affinity-purified polyclonal horse antibodies to human AFP were bound, the membranes were treated with or without rabbit immunoglobulins to human AFP, followed by overlaying with horseradish peroxidase-labeled goat anti-rabbit IgG for color development. Artificial complexes formed in vitro from human AFP and rabbit anti-AFP were clearly separated from free AFP by the agarose electrophoresis. The complexes were stained 20-40% as dark as the equivalent amount of free AFP by treatment with rabbit anti-AFP, and 10-20% as dark without the antibody treatment over a wide range of antigen-antibody ratios.

Developmental alterations in the alpha-fetoprotein sugar chain in maternal serum analyzed by lectin affinity electrophoresis.

Our purpose was to investigate developmental alterations of human alpha-fetoprotein (AFP) oligosaccharides in maternal serum by lectin affinity electrophoresis and to compare the AFP glycoforms in maternal serum with those in umbilical cord serum and amniotic fluid. AFP glycoforms were separated by affinity electrophoresis with concanavalin A (Con A), lentil lectin (LCA), erythroagglutinating phytohemagglutinin (E-PHA) and Allomyrina dichotoma lectin (allo A) and detected by sensitive antibody affinity blotting. In maternal serum, increased proportions of Con A-nonreactive AFP (AFP-C1), LCA strongly-reactive AFP (AFP-L3) and E-PHA-reactive AFP (AFP-P4 and AFP-P5) decreased gradually during the early gestational weeks. Allo A-nonreactive AFP (AFP-A1 and asialo-AFP) were found only in amniotic fluids during early gestational weeks. The percentages of these glycoforms at full term were almost the same among those body fluids. Since the glycoforms of maternal serum AFP were close to those of umbilical cord serum AFP, lectin-affinity electrophoretic analysis of maternal serum AFP may be useful for evaluating the developmental state of fetus by examining the nature of AFP sugar chain.

Developmental alterations of alpha-fetoprotein sugar chain in amniotic fluids analyzed by lectin affinity electrophoresis.

Affinity electrophoresis of human alpha-fetoprotein (AFP) in amniotic fluid from pregnant women between 6 to 42 weeks of gestation and in the serum of a yolk sac tumor was performed with concanavalin A (Con A), lentil lectin (LCA), erythroagglutinating phytohemagglutinin (E-PHA) and Allomyrina dichotoma lectin (allo A). Separated AFP bands were detected by sensitive antibody-affinity blotting. In the first trimester, amniotic fluid AFP showed elevated percentages of Con A-nonreacting AFP (AFP-C1) and LCA weakly-reacting AFP (AFP-L2) as previously reported. Additionally, high percentages of E-PHA strongly-reacting AFP (AFP-P5) and E-PHA-reacting AFP (AFP-P4) were observed. E-PHA-nonreacting AFP (AFP-P1), E-PHA weakly-reacting AFP (AFP-P3f), allo A-nonreacting AFP (AFP-A1) and asialo-AFP, AFP-A1 s, were present only in amniotic fluids from 6 to 17 weeks of gestation. With advancing gestation, percentages of AFP-C1, AFP-P4 and AFP-P5 decreased and AFP-L2, AFP-P3f, AFP-A1, and AFP-A1 s disappeared by the end of 18 weeks. The glycoforms of serum AFP of the yolk sac tumor resembled those of amniotic fluid AFP in the early gestational stages.

Effect of gegen-tang on painful gynecomastia in patients with liver cirrhosis: a brief report.

Four patients with liver cirrhosis and complaining of painful gynecomastia were treated with oral administration of Gegen-Tang (TJ-1). Pain disappeared in 3 patients in one week, and in one patient in 4 weeks. The size of gynecomastia did not change significantly on mammography, but palpable induration diminished or disappeared. The patients had been treated with Chaihu (saiko) group drugs for liver diseases, and TJ-1 was used in combination with these drugs. Serum levels of estrogen, progesterone, testosterone, and other sex hormones did not change significantly after TJ-1 treatment. These results suggest that TJ-1 could be used for the painful gynecomastia that is occasionally seen in cirrhotic patients.

Effect of Koso-san on globus pharyngeus.

Twenty-three patients with globus pharyngeus were treated with Koso-san (TJ-70) at a dose of 7.5 g/day for at least 14 days. Symptoms disappeared in 18 cases and improved in 3 cases, therefore, the effective rate was 91.3% (21/23). The symptoms disappeared on average within 13.5 days. Terasawa qi-stasis scores were significantly decreased after the treatment. TJ-70 might thus be a remedy for globus pharyngeus with qi-stasis.

Effect of niuche-shen-qi-wan on painful muscle cramps in patients with liver cirrhosis: a preliminary report.

Twelve patients with liver cirrhosis complaining of painful muscle cramps were treated with Niuche-Shen-Qi-Wan (TJ-107). Three patients were at the decompensated state. Muscle cramps disappeared in 4 weeks on the average after oral administration of TJ-107 in all 12 patients. During the period of TJ-107 administration, there was no significant improvement of hepatic function. One patient complained of mild epigastric discomfort after taking TJ-107, but there were no other adverse effects. Our results indicate that TJ-107 is useful for treatment of painful muscle cramps in cirrhotic patients.

[The effect of a mixture of tegafur with uracil (UFT) on hepatocarcinogenesis in rats induced by 3'-methyl-4-dimethylaminoazobenzene. I. The influence on production of alpha-fetoprotein].

We studied the effect of UFT (a mixture of Tegafur and uracil at a ratio of 1:4) on hepatocarcinogenesis and alpha-fetoprotein (AFP) production induced by 3'-methyl-4-dimethylaminoazobenzene (3'-MeDAB) in rats. By feeding rats with 3'-MeDAB diet, serum AFP started to appear from week 2-3 and increased until week 5-7, then declined until week 10-11. This phenomenon of a transient appearance of serum AFP is called "primary reaction". After week 11 hepatoma started to develop accompanied with an enormous elevation of serum AFP level until animals die, generally before week 25. Male Donryu strain rats fed a diet containing 0.06% 3'-MeDAB for 10 weeks, and then fed the normal diet. UFT was given by a stomach tube once a day for five days a week. Blood was collected from the tail vein every other week and serum AFP was measured by Rocket electrophoresis technique. When the administration of UFT (10 mg or 20 mg/kg/day) was started simultaneously with feeding 3'-MeDAB diet, the primary reaction was somewhat suppressed, and after week 11 development of hepatoma and elevation of AFP were markedly inhibited. The administration of UFT of a high dose (40 mg/kg/day) caused the death of animals in 7 weeks probably because of its toxicity. When the administration of UFT (10 mg or 20 mg/kg/day) was started after finish of feeding 3'-MeDAB diet for 10 weeks, the drug was not effective. These results show that if the administration of UFT was commenced at the same time with feeding 3'-MeDAB diet, the hepatocarcinogenesis and AFP production are strongly inhibited.

[Inhibitory effect of UFT on rat hepatocarcinogenesis induced by 3'-methyl-4-dimethylaminoazobenzene and phenobarbital promotion].

Inhibitory effect of UFT on hepatocarcinogenesis in rats induced by 3'-methyl-4-dimethylaminoazobenzene (3'-MeDAB) and phenobarbital (PB) promotion was studied. Donryu male rats were divided into four groups. Group A was fed a diet containing 0.06% 3'-MeDAB for 3, 5, or 7 weeks, and then fed normal diet for 2 weeks, subsequently received a diet containing 0.05% PB. Group B was given UFT (20 mg/kg/5 days a week) simultaneously with feeding 3'-MeDAB. Group C was given UFT simultaneously with feeding PB. Group D was given 3'-MeDAB alone. In all groups, the development of hepatocellular carcinoma was investigated 37 weeks later and the number and area per mm2 of induced glutathione S-transferase placental form (GST-P) positive foci were measured using an image processor. The number and area of GST-P positive foci in group B and group C were markedly decreased as compared with those in group A. These results seem to show that the administration of UFT inhibited the production of GST-P positive foci and that stronger inhibitory effect of UFT was observed by simultaneous administration of an initiator than by that of a promoter.