Biomedical subjects

S Hendrich

Publications and source records attributed to S Hendrich.

At least 37 records · Page 2Linked to original sources

PubMed1995

Fusarium proliferatum-fermented corn stimulates development of placental glutathione S-transferase-positive altered hepatic foci in female rats.

Groups of 8 6-w-old female Sprague-Dawley rats were initiated with 30 mg diethylnitrosamine (DEN)/kg. Control and initiated groups were fed a semipurified diet or diets supplemented with Fusarium proliferatum-contaminated corn to contain 20 or 50 mg fumonisin B1 (FB1)/kg. Histochemical staining for gamma-glutamyltransferase (GGT) and immunochemical staining for placental glutathione S-transferase (PGST), markers of altered hepatic foci (AHF), were performed on serial frozen hepatic sections. Gamma-glutamyltransferase -(+) AHF were not found in any group. Dosing with DEN significantly increased the number of PGST-(+) hepatocytes compared to the uninitiated groups. Groups fed F proliferatum-containing diets also had a significantly increased number of PGST-(+) AHF compared with those fed no F proliferatum. The volume percentage of liver occupied by PGST-(+) foci was significantly greater in the groups treated with DEN or F proliferatum. The number of PGST-(+) AHF/liver in the groups given DEN was also significantly greater than in the uninitiated groups. Fusarium proliferatum exposure also significantly increased the number of PGST-(+) AHF/liver. Feeding F proliferatum containing 20 mg FB1/kg promoted the development of DEN-initiated AHF in rats. Placental glutathione S-transferase was a more useful marker than GGT in detecting AHF produced by small amounts of F proliferatum mycotoxins fed after initiating dosing with DEN.

Animals↗

PubMed1994

S-thiolation and irreversible oxidation of sulfhydryls on carbonic anhydrase III during oxidative stress: a method for studying protein modification in intact cells and tissues.

S-thiolation of carbonic anhydrase III (CA III) in cultured rat hepatocytes under oxidative stress was studied by immunodetection on nitrocellulose blots of isoelectrofocusing gels. In cells treated with menadione, three S-thiolated forms of CA III were detected, whereas only two forms were observed in hepatocytes treated with t-butyl hydroperoxide. Two "nonreducible" oxidized forms of CA III were also detected on nitrocellulose blots. These forms increased with the amount of stress and were the only modified forms of CA III in buthionine sulfoxide-treated hepatocytes containing 10-fold less glutathione than control hepatocytes. These experiments support the concept that S-thiolation protects CA III from irreversible oxidation during oxidative stress. Partly and fully S-thiolated forms of CA III were easily detected in both male and female hepatocytes by the immunoblotting method, although female cells contained 15-fold less CA III than did male liver. S-thiolated forms of CA III were also detected in rat skeletal muscle and heart showing the utility of this method for determining the effect of oxidative stress on specific S-thiolatable protein in several tissues in vivo.

Animals↗

PubMed1994

Protein S-thiolation in hepatocytes stimulated by t-butyl hydroperoxide, menadione, and neutrophils.

In order to examine potentially important S-thiolated proteins, 35S-labeled hepatocytes were exposed to oxidative stress. A similar group of S-thiolated proteins including carbonic anhydrase III was observed in cells treated with t-butyl hydroperoxide, menadione, or stimulated neutrophils. The radioactive thiols bound to hepatocyte proteins were identified by HPLC and more than 85% was glutathione. In menadione-treated hepatocytes, proteins were gradually S-thiolated over 30 min and 25% of the cellular glutathione pool became protein-bound. In t-butyl hydroperoxide-treated cells, S-thiolation was more transient and 11% of the glutathione was protein-bound. Neutrophil-treated hepatocytes had nearly the same amount of protein S-thiolation (8% after 25 min). Two major proteins that were S-thiolated in untreated hepatocytes did not increase during any form of oxidative stress. In neutrophil-treated hepatocytes protein S-thiolation was not accompanied by either formation of glutathione disulfide or a measurable change in the total amount of glutathione. In both t-butyl hydroperoxide-and menadione-treated cells there was extensive formation of glutathione disulfide and in menadione-treated cells a significant increase in the total hepatocyte glutathione pool was observed. This result suggests that protein S-thiolation may occur by mechanisms that do not result from thiol/disulfide exchange between glutathione disulfide and protein sulfhydryls. It is suggested that a thiyl radical intermediate is important in neutrophil-mediated protein S-thiolation.

Animals↗

PubMed1994

Vitamin E deficiency increases serum thromboxane A2, platelet arachidonate and lipid peroxidation in male Sprague-Dawley rats.

This study was designed to determine whether dietary linoleate and all-rac-alpha-tocopheryl acetate (vitamin E) interact to affect serum thromboxane A2 (TXA2) and prostacyclin (PGI2) status and therefore, thrombogenic potential. 6 groups of 12 weanling male Sprague-Dawley rats were fed semipurified diets containing 11 or 18% of energy from linoleate and 0, 100 or 5000 mg vitamin E/kg diet for 10 weeks. Platelet and serum alpha-tocopherol concentrations increased logarithmically with increasing dietary vitamin E. Serum TXA2, measured as TXB2, platelet arachidonate and thiobarbituric acid reactive substances were significantly greater in the vitamin E deficient groups than in groups receiving vitamin E (p < 0.05). Serum PGI2 levels, determined as 6-keto-PGF1 alpha, were not affected by diets. No interaction was found between dietary linoleate and vitamin E. However, vitamin E supplementation produced significantly less serum TXB2 than did vitamin E deficient diets (p < 0.05). Vitamin E deficiency may be prothrombogenic by increasing platelet arachidonate, lipid peroxidation and serum TXA2 levels while vitamin E supplementation at levels used in this study may decrease such effects.

6-Ketoprostaglandin F1 alpha↗

PubMed1994

Daidzein is a more bioavailable soymilk isoflavone than is genistein in adult women.

Soybean isoflavones are proposed to be anticarcinogenic, but their effective doses have not been established. To study the bioavailability of soybean isoflavones for humans, 12 young adult women received single doses of 0.7, 1.3 and 2.0 mg isoflavones/kg body wt in soybean milk as part of a liquid diet. Plasma, urine and fecal isoflavones were measured by reverse-phase HPLC. Average 24-h urinary recoveries of daidzein and genistein were approximately 21% and 9%, respectively, at all three doses. Urinary recovery of daidzein was significantly greater than that of genistein (P < 0.001). Total fecal excretion of isoflavones was only 1-2% of the ingested amount. Plasma total isoflavone concentration was significantly increased to 4.4 +/- 2.5 mumol/L at 6.5 h after a dose of 2.0 mg/kg. The plasma concentrations of daidzein and genisten were approximately equal. Twenty-four hours after dosing, both plasma and urine isoflavone concentrations were nearly nil. Although soybean milk isoflavones seem to be 85% degraded in the intestine, the bioavailability, especially of daidzein, may be sufficient to exert some health-protective effects.

Administration, Oral↗

PubMed1994

Defining food components as new nutrients.

When obtained form a usual diet, a food component that sustains or enhances physiological functions and/or prevents diseases is a nutrient. Isoflavones, tocotrienols, and carotenoids are candidate nutrients which may be of health benefit to humans by inhibiting cancer development and reducing risk of atherosclerosis. The amounts of some of these candidate nutrients in food are known. A carotenoid data base has been developed. Isoflavone content of soy foods ranges from 0.1 mg/g (soymilk) to 2.5 mg/g (soy protein isolate). Human bioavailability studies have also been performed with these candidate nutrients. For example, in young adult females fed a single meal containing soy milk, isoflavones were cleared from urine within 24 h after feeding, with about 15-20% of the total dose accounted for in urine and feces. The two major soy isoflavones, genistein and daidzein, differ in bioavailability, with daidzein being more readily excreted in urine. Isoflavones, tocotrienols, and carotenoids meet several criteria for classification as nutrients. But after appropriate animal testing, food analyses, and availability studies have been performed, human health-protective efficacy must be proven in long-term feeding trials, in order for potential health-enhancing food components to be classified as nutrients.

Biological Availability↗

PubMed1993

Selenium deficiency suppresses the S-glutathiolation of carbonic anhydrase III in rat hepatocytes under oxidative stress.

To examine the modification of reactive sulfhydryls of carbonic anhydrase III (CA III), hepatocytes were prepared by collagenase perfusion from Se deficient and Se-adequate male Sprague-Dawley rats. After 24 h in culture, hepatocytes were treated for 15-30 min with one of two oxidative stressors, t-butyl hydroperoxide (t-BuOOH) or menadione. Modification of CA III was measured by isoelectric focusing/immunoblotting. Formation of glutathione disulfide (GSSG) during oxidative stress was markedly less in hepatocytes of Se-deficient rats than in those of Se-adequate rats. During treatment with t-BuOOH, GSSG formation in hepatocytes from Se-adequate rats reached a maximum at 3 min, and then GSSG was gradually reduced to glutathione. After menadione treatment, intracellular GSSG irreversibly increased in hepatocytes of Se-adequate rats but not in those of Se-deficient rats. A modification of CA III that was reversible by dithiothreitol treatment concurred with the formation of GSSG during treatment with either t-BuOOH or menadione. Although modification of CA III occurred in hepatocytes from Se-deficient rats, the extent of modification was significantly less than in Se adequacy, and the modification was less reversible by dithiothreitol than in hepatocytes from Se-adequate rats. Selenium deficiency may be useful in examining the importance of modification of specific proteins subjected to oxidative stress.

Analysis of Variance↗

PubMed1992

Gender and dietary fat affect alpha-tocopherol status in F344/N rats.

For four weeks, groups of eight male and eight female F344/N rats were fed diets containing 15.5, 20, 30 or 40% of energy (en%) as fat. The fat was composed of corn oil and beef tallow with 9 en% from linoleate in all diets. Females had greater mean hepatic alpha-tocopherol levels, whereas males had greater plasma alpha-tocopherol and cholesterol concentrations. In males, the plasma ratio of alpha-tocopherol/cholesterol was significantly greater than in females (P < 0.05). Plasma alpha-tocopherol increased with increasing en% fat (r = 0.51, P < 0.001) in both sexes, but dietary fat did not alter hepatic alpha-tocopherol levels. These results suggest that plasma alpha-tocopherol may serve as a biomarker of total dietary fat intake and that in F344/N rats gender differences affect alpha-tocopherol and cholesterol status.

Animals↗

PubMed1992

Sex and dietary fat modulate hepatic prostaglandin F2 alpha in F344/N rats.

The study was designed to determine whether sex and fat calories altered hepatic prostaglandin (PG) F2 alpha status; a factor which may reflect susceptibility to cancer development. For 4 weeks, groups of 8 male and 8 female F344/N rats were fed diets with 9% of energy (en%) from linoleate and 15.5, 20, 30 or 40 en% fat. Females had greater hepatic stearate, arachidonate and PGF2 alpha whereas males had greater hepatic myristate, palmitate and oleate. Females also had greater plasma stearate levels. Greater hepatic arachidonate may have stimulated PG production in females. Hepatic oleate increased and hepatic palmitate decreased with increasing en% fat (p < 0.05). Hepatic stearate was greater and hepatic linoleate less when 40 en% fat was fed compared with other levels of dietary fat (p < 0.05). Plasma oleate was greater at 30 or 40 en% fat than at lower levels of fat, whereas plasma linoleate was less at 40 en% than at 15.5% en% fat. The ability of a 30 en% fat diet, containing equal proportions of linoleate and oleate, to suppress hepatic PG production may be related to the effects of dietary fat content and composition on plasma fatty acid profiles. Because suppressed PG production has been linked with suppression of cancer development, dietary recommendations to consume 30 en% fat with a P:M ratio of 1:1 may be cancer-protective.

Animals↗

PubMed1992

Suppression of hepatic prostaglandin F2 alpha in rats by dietary alpha-tocopherol acetate is independent of total hepatic alpha-tocopherol.

Groups of eight weanling female F344/N rats were fed semipurified diets that supplied 0, 50, 500, 5000, or 15,000 mg alpha-tocopherol acetate/kg diet, with and without 0.05% phenobarbital (PB) for 9 weeks. Both plasma and hepatic alpha-tocopherol levels, measured by HPLC, strongly correlated with alpha-tocopherol intake (r greater than 0.73, p less than 0.0001). Phenobarbital both depleted hepatic alpha-tocopherol and increased plasma alpha-tocopherol significantly. Although treatment with PB for 9 weeks significantly increased GST activity, PB did not affect hepatic prostaglandin (PG)F2 alpha status, as determined by radioimmunoassay. PGF2 alpha was significantly greater (by 52%) in rats fed no alpha-tocopherol than in rats fed 15,000 mg alpha-tocopherol acetate/kg diet. Hepatic PGF2 alpha status was correlated inversely but weakly with dietary alpha-tocopherol (r = -0.24, p less than 0.05). Hepatic PGF2 alpha status was not correlated with hepatic or plasma alpha-tocopherol status. This finding suggests either that there is a small depletion-resistant subcellular alpha-tocopherol pool which regulates PGF2 alpha production or that alpha-tocopherol alters PGF2 alpha production in vivo by an indirect mechanism.

Animals↗

PubMed1992

Ovariectomy promotes the growth of altered hepatic foci after withdrawal and reintroduction of phenobarbital during hepatocarcinogenesis in rats.

Female F344/N rats were given 70% partial hepatectomies and intubated with diethyl-nitrosamine (DEN, 10 mg/kg) 24 hours later. They were fed a cereal-based diet, NIH-07 (NIH) + 0.05% phenobarbital (PB) for 6 months, at which time NIH + PB was withdrawn and the rats were ovariectomized (OV) or sham-operated (SH). Groups of 7-10 rats were fed a semipurified diet (AIN-76) for 1 or 2 months after withdrawal of NIH + PB, or NIH + PB for 2 months, or AIN-76 diet for 1 month and subsequently NIH + PB for 1 month. Placental glutathione S-transferase (PGST)- and gamma-glutamyltransferase (GGT)-positive (+) altered hepatic foci (AHF) were analysed by quantitative stereology. Ovariectomy stimulated growth of AHF after withdrawal and reintroduction of NIH + PB. AHF, especially PGST+ AHF, continued to regress throughout the PB withdrawal period in rats fed AIN-76 diet. In most studies of chemical hepatocarcinogenesis, females have been shown to develop a greater volume of AHF than males. In our study, however, ovariectomy stimulated the growth of AHF after withdrawal and reintroduction of PB. Because AHF occurring spontaneously in male rats develop more rapidly than in female rats, the greater rate of growth of AHF in OV female rats may reflect a similar mechanism.

Animals↗

PubMed1991

Identification of an abundant S-thiolated rat liver protein as carbonic anhydrase III; characterization of S-thiolation and dethiolation reactions.

An S-thiolated 30-kDa protein has been purified from rat liver by two steps of ion-exchange chromatography. This monomeric protein has two "reactive" sulfhydryls that can be S-thiolated by glutathione (form a mixed disulfide with glutathione) in intact liver. The protein has been identified as carbonic anhydrase III by sequence analysis of tryptic peptides from the pure protein. The two "reactive" sulfhydryls on this protein can produce three different S-thiolated forms of the protein that can be separated by isoelectric focusing. Using this technique it was possible to study the S-thiolation and dethiolation reactions of the pure protein. The reduced form of this protein was S-thiolated both by thiol-disulfide exchange with glutathione disulfide and by oxyradical-initiated S-thiolation with reduced glutathione. The S-thiolation rate of this 30-kDa protein was somewhat slower than that of glycogen phosphorylase b by both S-thiolation mechanisms. The S-thiolated form of this protein was poorly dethiolated (i.e., reduced) by glutathione, cysteine, cysteamine, or coenzyme A alone. Enzymatic catalysis by two different enzymes (glutaredoxin and thioredoxin-like) greatly enhanced the dethiolation rate. These experiments suggest that carbonic anhydrase III is a major participant in the liver response to oxidative stress, and that the protein may be S-thiolated by two different non-enzymatic mechanisms and dethiolated by enzymatic reactions in intact cells. Thus, the S-thiolation/dethiolation of carbonic anhydrase III resembles glycogen phosphorylase and not creatine kinase.

Amino Acid Sequence↗

PubMed1991

Phenobarbital increases rat hepatic prostaglandin F2 alpha, glutathione S-transferase activity and oxidative stress.

Eight-week-old female F344/N rats were fed 3.0 or 6.0% of calories (kcal%) as linoleate with or without 0.05% phenobarbital (PB) for 35 days. PB treatment increased glutathione S-transferase (GST) activity by 80% and prostaglandin (PG) F2 alpha levels 4-fold (p less than 0.05). PB decreased hepatic alpha-tocopherol significantly. Hepatic linoleate was decreased by PB in rats fed 6 kcal% but not 3 kcal% linoleate. Increased dietary linoleate had no significant effect on hepatic PGF2 alpha or alpha-tocopherol levels or GST activity. This study suggests that PB hepatotoxicity and tumor-promoting ability may be mediated, at least in part, by PGF2 alpha. PB's effect on PGF2 alpha could be a result of both GST-mediated prostaglandin synthesis and oxidative stress. The removal of significant amounts of hepatic alpha-tocopherol during oxidative stress induced by PB might diminish endogenous inhibition of hepatic PG synthesis by a-tocopherol.

Animals↗

PubMed1991

Effects of alpha-tocopherol, phenobarbital, and butylated hydroxyanisole during promotion of diethylnitrosamine-initiated rat hepatocarcinogenesis.

The promotion-suppressing ability of two antioxidants was measured to determine the role of oxidative stress in hepatocarcinogenesis. Four-day-old female F344/N rats were dosed with diethylnitrosamine (10 mg/kg). After weaning, they were fed semipurified diets with and without 500 ppm alpha-tocopherol, or the same two diets containing 500 ppm phenobarbital, or 5,000 ppm butylated hydroxyanisole (BHA) for 3 or 11 months. By 11 months, phenobarbital-fed groups had eaten 30% more than other groups did (p less than 0.05), suggesting a role for increased caloric intake in phenobarbital promotion. Phenobarbital and BHA significantly reduced body weights and increased liver weights compared with control rats. After three months, alpha-tocopherol significantly suppressed mean volume of placental glutathione S-transferase (PGST)-positive altered hepatic foci (AHF), regardless of xenobiotic treatment. Phenobarbital increased and BHA decreased the numbers of AHF compared with those of the control group. After 11 months, mean focal volume was significantly suppressed by BHA compared with that of the control group, and phenobarbital increased the total volume of AHF [PGST-positive plus gamma-glutamyltransferase (GGT)-positive AHF] compared with rats fed either control or BHA diets. BHA treatment also increased hepatic glutathione levels by 40% compared with control and rats fed phenobarbital. In conclusion, alpha-tocopherol had only a slight, early effect to suppress promotion of hepatocarcinogenesis. BHA suppressed some indices of promotion at both times and increased hepatic glutathione; however, BHA's toxicity (which suppressed body weight) may also be a factor in its supposable promotion-inhibitory effects.

Analysis of Variance↗

PubMed1990

Quantitative stereological analysis of the effects of age and sex on multistage hepatocarcinogenesis in the rat by use of four cytochemical markers.

Altered hepatic foci (AHF) were analyzed by quantitative stereology on frozen serial sections stained sequentially for gamma-glutamyltranspeptidase (GGT), canalicular adenosine triphosphate (ATPase), glucose-6-phosphatase (G6Pase), and the placental isoenzyme of glutathione S-transferase (GST). Livers for these analyses were obtained from both male and female rats of different ages which had been subjected to initiation with a nonnecrogenic dose of diethylnitrosamine following a 70% partial hepatectomy with subsequent phenobarbital (PB) feeding. Different combinations of these four marker alterations (from single marker to four-marker combinations) were used to analyze the data, and the results were compared for their ability to detect AHF. In rats on the above protocol, GST was the single most effective marker, exhibiting a high sensitivity for scoring both number and volume of foci. There was a high degree of overlap with GGT. The combination of the four different markers, GST/GGT/ATPase/G6Pase, scored 80% more foci in number and 60% more in volume than the routinely used GGT/ATPase/G6Pase method. When all four markers were used to score AHF, PB promotion was equally effective in both sexes at weaning and at 6 months of age, but at 1 year of age males showed a dramatic reduction in the effectiveness of PB as a promoting agent, both for number and volume percentage of liver occupied by AHF. On the other hand, initiation was more effective in the male at weaning and at 6 months of age, although by the 12-month point no distinction between the sexes could be made. When only GGT was used as a marker, promotion by PB appeared to be markedly less effective in males than in females at all ages. In the absence of PB administration, both the number and volume fraction of AHF in the livers of both males and female increased with age. Likewise, both the number of AHF per liver and their volume fractions increased with age in both sexes when uninitiated animals were fed PB, although only after a 6-month lag in females. These experiments demonstrate that the stages of initiation and promotion in hepatocarcinogenesis in the rat as monitored by the number and volume percentage occupied of AHF are altered by both the age and the sex of the animal. The combination of GGT and GST identified all AHF scored by the GST/GGT/ATPase/G6Pase set of markers and thus may be the most efficient combination of markers of AHF resulting from promotion by PB.

Adenosine Triphosphatases↗

PubMed1989

Regulation of the expression of some genes for enzymes of glutathione metabolism in hepatotoxicity and hepatocarcinogenesis.

The reversible stage of tumor promotion, which follows the stage of initiation and precedes that of progression in multistage carcinogenesis, is a unique example of reversible toxicity in biological systems. In order to study the molecular mechanisms involved in the action of promoting agents during this stage, the regulation of the expression of genes for two enzymes of glutathione metabolism, gamma-glutamyl transpeptidase (GGT) and the placental isozyme of glutathione S-transferase (GST-P), was studied under several different conditions of promotion during multistage hepatocarcinogenesis in the rat. Promotion by phenobarbital caused an increased expression of both of these genes in altered hepatic focal lesions, although this was somewhat more variable in the case of the GGT gene. C.I. Solvent Yellow 14, an industrial dye, served as an effective promoting agent. Feeding this dye resulted in a dramatic increase in the expression of GST-P, but not that of GGT in altered hepatic foci. Factors in crude, cereal-based diets inhibited the stage of promotion by diethylnitrosamine, but enhanced promotion by phenobarbital in a synergistic manner. In contrast, at least one purified diet had the converse effect during this stage. The mRNA levels of GST-P were uniformly elevated dramatically in reversible nodules and neoplasms of rat liver that had been induced by diethylnitrosamine and phenobarbital promotion. In contrast, the level of GGT mRNA was somewhat variable, with an occasional neoplasm exhibiting almost a background level of expression of this gene. Therefore, the altered regulation of multiple genes in hepatocytes during the stage of promotion can vary with the promoting agent itself; this process may be related to the heterogeneous gene expression seen in hepatic neoplasms. A possible role for specific DNA sequences in the 5' flanking regions of such genes is considered. In addition, a cDNA clone to the mRNA of human liver GGT was isolated and sequenced. The homology of the coding sequence of the human liver GGT mRNA to that of rat kidney GGT mRNA was striking.

Animals↗

PubMed1989

A semipurified diet that suppresses phenobarbital promotion of hepatocarcinogenesis in the rat.

Six groups of F344/N female rats were fed either a modified AIN-76 diet (20% casein, 5% corn oil, 65% cornstarch, 5% cellulose) (AIN) or a diet formulated by Dr. M. Pariza (PD) (30% casein, 10% partially hydrogenated corn oil, 40% sucrose, 15% cornstarch) beginning four days before 70% partial hepatectomy. One day after the surgery, one group fed each diet was intubated with 10 mg/kg diethylnitrosamine (DEN). One week later, these groups plus one control group fed each diet were given 0.05% phenobarbital in the diet for 6 or 14 months. After the rats were killed, blocks of liver tissue were frozen on dry ice and stored at -70 degrees C. Three frozen serial sections were stained for gamma-glutamyltransferase, ATPase, and glucose-6-phosphatase. Numbers and volume of altered hepatic foci (AHF) were analyzed by stereological techniques. After 14 months of feeding these regimens, rats initiated with DEN and fed the AIN + PB had significantly greater numbers and a higher percent volume of the liver of most phenotypes of AHF than all other groups, including those fed PD + PB following initiation with DEN. The numbers of AHF exhibiting more complex phenotypes (i.e., scored by more than one marker) remained unaltered between 6 and 14 months. These findings indicate that the effectiveness of PB as a promoting agent in multistage hepatocarcinogenesis is significantly altered when fed with two different diets of known composition. Therefore, dietary composition can be a significant factor in studies of the stage of promotion in hepatocarcinogenesis.

Animals↗

PubMed1988

Dietary effects on initiation and promotion of hepatocarcinogenesis in rat.

Female F344/N rats were initiated with DEN (10 mg/kg) 24 h after a 70% partial hepatectomy. Groups of 10 rats were fed (a) AIN, group-1; (b) PD, group-2; or (c) NIH, group-3, for 1 week after initiation and were then fed NIH plus the promoting agent PB at a level of 0.05% in the diets for 6 months. Other groups were fed NIH for 1 week after initiation and then NIH without PB (group-4), AIN + PB (group-5), AIN without PB (group-6), PD + PB (group-7), or PD without PB (group-8) for 6 months. The numbers and volume percentages of AHF were quantified by stereologic methods from frozen serial sections, stained consecutively for GGT, ATPase, and G6Pase. For the groups fed different diets during the 1st week after initiation, the numbers and volume of AHF were significantly greater in group-2 than in groups 1 or 3. The numbers of AHF were significantly less in group-3 than in group-1. The numbers and volume of AHF were significantly greater in groups fed PB during the promotion phase, except in the case of group-7, whose focal volume did not differ from groups 6 or 8. Group-3 had significantly greater numbers of AHF than groups 5 and 7. These findings can be explained by the hypothesis that the NIH diet contained factors that acted synergistically with PB to enhance tumor promotion. The mean focal volume of both GGT positive and ATPase negative foci was significantly greater in group-5 than in all other groups; this indicates that the AIN + PB regimen selectively promoted the growth of a subpopulation of AHF. These findings show that alterations in the composition of diets fed during hepatocarcinogenesis significantly alter the effects of specific chemical agents acting during the stages of initiation and promotion in hepatocarcinogenesis.

Animals↗