White and wholemeal flours from wheats of low and higher apparent metabolizable energy differ in their nutritional effects in rats.

Wheats used for feeding poultry differ considerably in the ratio of soluble to insoluble non-starch polysaccharides (NSP) and apparent metabolizable energy (AME). We have examined effects of whole and white flour from a wheat of low (12.02 MJ/kg of dry matter) and high (14.52 MJ/kg of dry matter) AME in rats fed a cholesterol-free diet. NSP concentrations were higher in whole flour from the low AME wheat but similar in both white flours. In contrast to chickens, food intake and body weight gain of rats were unaffected by diet. Plasma cholesterol concentrations were lower in rats fed whole wheat and low AME wheat. Cecal bile acids and neutral sterol pools were larger in rats fed whole wheat but were unaffected by wheat type. Negative correlations were found between digesta steroid pools and plasma cholesterol, consistent with greater fecal steroid excretion. Cecal digesta was greater in rats fed whole wheat and low AME wheat. Digesta pH was lower in rats fed whole wheat, but there was a significant interaction between wheat and flour type with lower pH in rats fed low AME white flour. Total volatile fatty acids (VFA) and acetate and butyrate pools were larger in rats fed whole wheat than in those fed white flour. Total and individual VFA pools were larger in rats fed low AME flours than in those fed high AME flours, apparently due to greater cecal starch fermentation in the former. Factors affecting wheat AME in chickens affect important metabolic variables in rats and may have similar actions in other species including humans.

Fecal numbers of bifidobacteria are higher in pigs fed Bifidobacterium longum with a high amylose cornstarch than with a low amylose cornstarch.

Twelve young male pigs consumed a purified diet containing wheat bran as fiber source. Starch provided 50% of total daily energy either as a low amylose cornstarch or as a high amylose (amylomaize) starch. The pigs were given a supplement of a freeze-dried probiotic organism (Bifidobacterium longum CSCC 1941). A block crossover design was used so that at any one time two groups of three pigs consumed either the high or low amylose cornstarch without probiotic and a further two groups of three pigs consumed either high or low amylose cornstarch with probiotic. Neither food intake nor body weight gain was affected by diet. Fecal output was higher when pigs were fed the high amylose cornstarch, but moisture content was unaffected. Fecal concentrations and excretion of total volatile fatty acids were higher when pigs were fed the high amylose cornstarch. Concentrations of acetate were unaffected by dietary starch, but those of propionate and butyrate were higher when the high amylose cornstarch was consumed. Fecal excretion of all three acids was higher during high amylose cornstarch feeding. Bifidobacteria were detected in the feces only when pigs were fed Bifidobacterium longum. Fecal bifidobacteria counts (expressed per gram of wet feces) and their daily fecal excretion were higher when pigs were fed high amylose cornstarch. Feeding the probiotic did not alter fecal starch or volatile fatty acids. None of the variables studied was affected by the order of feeding of starch or probiotic. The data show that a high amylose starch acts as a prebiotic in promoting the fecal excretion of probiotic organisms.

A high amylose (amylomaize) starch raises proximal large bowel starch and increases colon length in pigs.

Young male pigs consumed a diet of fatty minced beef, safflower oil, skim milk powder, sucrose, cornstarch and wheat bran. Starch provided 50% of total daily energy either as low amylose cornstarch, high amylose (amylomaize) cornstarch or as a 50/50 mixture of corn and high amylose starch. Neither feed intake nor body weight gain as affected by dietary starch. Final plasma cholesterol concentrations were significantly higher than initial values in pigs fed the 50/50 mixture of corn and high amylose starch. Biliary concentrations of lithocholate and deoxycholate were lower in pigs fed high amylose starch. Large bowel length correlated positively with the dietary content of high amylose starch. Concentrations of butyrate in portal venous plasma were significantly lower in pigs fed high amylose starch than in those fed cornstarch. Neither large bowel digesta mass nor the concentrations of total or individual volatile fatty acids were affected by diet. However, the pool of propionate in the proximal colon and the concentration of propionate in feces were higher in pigs fed amylose starch. Concentrations of starch were uniformly low along the large bowel and were unaffected by starch type. In pigs with cecal cannula, digesta starch concentrations were higher with high amylose starch than with cornstarch. Electron micrographic examination of high amylose starch granules from these animals showed etching patterns similar to those of granules obtained from human ileostomy effluent. It appears that high amylose starch contributes to large bowel bacterial fermentation in the pig but that its utilization may be relatively rapid.

Plasma lipids and large bowel volatile fatty acids in pigs fed on white rice, brown rice and rice bran.

Adult male pigs were fed on a diet containing (% of energy) fat 25 starch 55 from white rice and providing 20 g fibre/pig d (diet WR). In two other groups rice bran was added to the diet to provide 43 g fibre/d. One group received the diet unmodified (diet RB), but in another (diet RO) heat-stabilized unrefined rice oil replaced the palm oil. In a further group brown rice replaced white rice and provided 37 g fibre/pig per d (diet BR). Plasma cholesterol concentrations were similar with diets WR, RB and BR. With diet RO the concentration was significantly lower than with diets WR and BR but was not different from diet RB. Plasma high-density-lipoprotein-cholesterol and plasma triacylglycerols were unaffected by diet. In all groups, digesta mass rose from the caecum to the proximal colon but fell in the distal colon. Diet WR gave the lowest digesta mass while diet BR gave a significantly higher mass along the large bowel length. RB- and RO-fed pigs had equal masses of digesta which were intermediate between BR- and WR-fed pigs at all sampling sites. Pools of individual and total volatile fatty acids (VFA) in the proximal large bowel were unaffected by diet. Pools of total and individual VFA in the median and distal colon were lowest with diets WR and RB and significantly higher with diet BR. In these regions of the colon pools of acetate in RO-fed pigs did not differ from those in the BR-fed group but were higher than in other groups. However, pools of propionate and butyrate with the RO diet were significantly lower than with diet BR and the same as with diets WR and RB. Portal venous VFA concentrations were unaffected by diet. The higher large bowel digesta masses and VFA with diet BR may reflect the escape of starch from the small intestine.

White wheat flour lowers plasma cholesterol and increases cecal steroids relative to whole wheat flour, wheat bran and wheat pollard in rats.

Plasma cholesterol concentrations were significantly lower in rats fed a cholesterol-free diet containing white wheat flour than those fed the diet with whole wheat or wheat bran. Concentrations of total bile acids and neutral sterols in cecal digesta were significantly higher in rats fed wheat flour than in those fed whole wheat, wheat pollard or wheat bran. Digesta bile acids and neutral sterol pools correlated negatively with plasma cholesterol, indicating that excretion was regulating plasma concentration. Total cecal volatile fatty acid (VFA) concentrations were unaffected by diet but cecal propionate was higher and butyrate lower in rats fed wheat flour than in those fed whole wheat. Cecal digesta butyrate concentrations correlated negatively with the cholesterol metabolite, coprostanol, and with secondary bile acids. Cecal propionate correlated negatively with plasma cholesterol concentration, but butyrate correlated equally positively, suggesting these VFA were indicators rather than regulators of altered cecal steroid metabolism. Effects of white wheat flour on steroid metabolism and cecal VFA resemble those of oat bran and support the observation that wheat flour might be hypocholesterolemic in humans.

Dietary fat and fiber alter large bowel and portal venous volatile fatty acids and plasma cholesterol but not biliary steroids in pigs.

Male pigs were fed a low fiber beef diet (control) or that diet with additional fiber either as wheat bran, oat bran or baked beans. Total large bowel digesta and volatile fatty acid (VFA) pools were highest in pigs fed the diet with baked beans, intermediate in those fed the diets with oat bran and wheat bran and lowest in those fed the control diet. In all groups digesta mass and total VFA pools rose from the cecum and then fell to the distal colon, and incremental effects of diet were the same at all sampling sites. For acetate and propionate pools there was a significant interaction between diet and anatomical site, but data conversion to logarithms abolished this interaction, indicating that all dietary effects were proportionately the same across sections. Consumption of the diets with wheat bran, oat bran and baked beans increased the total large bowel butyrate pool compared with consumption of the control diet. Digesta H+ concentrations fell along the large bowel and correlated positively with VFA concentrations in the median colon. Portal venous VFA concentrations correlated with VFA in the proximal colon only. Plasma cholesterol and biliary steroids were unrelated to portal venous propionate concentrations.

Fish oil and oat bran in combination effectively lower plasma cholesterol in the rat.

Male rats were fed a semi-purified diet containing oat bran or wheat bran with or without a marine fish oil to investigate the effects of such combinations on lipid metabolism. Oat bran alone and wheat bran plus fish oil gave lower plasma cholesterol concentrations than wheat bran alone while oat bran plus fish oil gave the lowest. Oat bran increased plasma triacylglycerols compared with wheat bran but oat bran plus fish oil gave concentrations similar to those seen with wheat bran plus fish oil. Oat bran gave higher hepatic cholesterol synthesis rates and a higher activity of 3-hydroxy-3-methylglutaryl coenzyme A (HMG-CoA) reductase compared to wheat bran. The addition of fish oil to either bran diet decreased cholesterol synthesis but HMG CoA reductase activity was not reduced. Oat bran increased hepatic acyl coenzyme A:cholesterol acyl transferase (ACAT) activity and increased the ratio of esterified to unesterified cholesterol in hepatic microsomal membranes compared with wheat bran. Fish oil decreased hepatic LDL receptor activity and increased HDL binding activity when added to the wheat bran diet but these effects were not seen with oat bran. Oat bran also had no effect on hepatic lipoprotein receptor activity compared with wheat bran. These results show that fish oil and oat bran have complementary cholesterol lowering effects in the rat.

Effects of solvent extraction on the hypocholesterolaemic action of oat bran in the rat.

In adult male rats fed on a cholesterol-free synthetic diet, plasma cholesterol concentrations were lowest with oat bran, intermediate with cellulose and highest with wheat bran. Plasma triacylglycerols (TAG) were similar with wheat bran and cellulose but higher with oat bran. The concentrations and pools of caecal volatile fatty acids (VFA) were lowest with cellulose and equally higher with oat bran and wheat bran. Plasma VFA concentrations in the hepatic portal vein reflected those in caecal digesta and were unrelated to plasma cholesterol. Feeding oat bran after extraction with n-pentane gave plasma cholesterol concentrations similar to that found with wheat bran. Reconstitution of oat bran with extracted lipids did not restore the cholesterol-lowering effect. Addition of the extracted material to a wheat-bran diet had no effect on plasma cholesterol. Plasma TAG were higher with the oat bran and reconstituted-oat-bran diets than with wheat-bran or cellulose diets. However, extracted oat bran + safflower oil gave similar TAG concentrations to that with wheat bran. These extractions and additions did not change caecal bile acid or neutral sterol concentrations. Effects of these diets on plasma cholesterol were unrelated to their tocotrienol or tocopherol content. Addition of n-pentane to oat bran followed by evaporation of solvent gave plasma cholesterol concentrations that were significantly higher than untreated oat bran but lower than similarly treated wheat bran. It is concluded that oat bran affects cholesterol metabolism through a pentane-soluble component as well as non-starch polysaccharides. It appears that the activity of this lipid is not transferable by simple addition of the solvent extract to the whole diet.

Modulation of the hypolipidemic effect of fish oils by dietary fiber in rats: studies with rice and wheat bran.

Adult male rats were fed a purified diet containing rice bran or wheat bran with or without a marine fish oil to investigate the possible effects of such dietary combinations on lipid metabolism. Plasma and hepatic triacylglycerols and hepatic lipogenesis were lowered significantly by feeding fish oil with rice bran but not with wheat bran. Plasma cholesterol and hepatic cholesterol synthesis were significantly lower in animals fed fish oil with either bran. Liver microsomal free cholesterol was significantly lower in rats fed rice bran alone than in all other groups. Hepatic low density lipoprotein (LDL) receptor activity was significantly higher in the two groups fed rice bran than in the two groups fed wheat bran. Fish oil significantly decreased hepatic LDL receptor activity and increased hepatic high density lipoprotein (HDL) binding activity with wheat bran but had no significant effects on these parameters when added to the rice bran diet. However, when the data for all groups were pooled, there was a significant negative correlation between hepatic HDL binding activity and LDL receptor activity. Cecal volatile fatty acids were significantly higher in rats fed rice bran, were unaffected by adding fish oil to either bran diet and did not appear to mediate any of the effects of the brans and fish oil on plasma lipids and hepatic lipid metabolism. The combination of rice bran plus fish oil therefore appears to have more beneficial effects on lipid metabolism than wheat bran plus fish oil.

Hypocholesterolaemic effects of dietary propionate: studies in whole animals and perfused rat liver.

In adult male rats fed a non-purified diet supplemented with 5% sodium propionate, plasma cholesterol concentrations were significantly depressed. Although liver cholesterol was increased by feeding propionate, rates of hepatic cholesterol and fatty acid synthesis were unchanged. Tissue concentrations and rates of synthesis of cholesterol were also unaffected by dietary propionate in stomach, small intestine and caecum. Concentrations of propionate in hepatic portal venous plasma were raised by feeding the supplemented diet but the increase was low in comparison to the dietary intake. Examination of the gut contents revealed concentrations of total volatile fatty acids (VFA) of 19 mumol/ml in the stomach contents of control rats and 148 mumol/ml (of which propionate contributed 116 mumol/ml) in those fed the supplemented diet. Duodenal and ileal concentrations of VFA were very low and were only slightly raised in the propionate-fed rats while caecal VFA were the same in both groups with a combined mean of 159 mumol/ml. These data indicate that in the rat, the absorption of dietary propionate appears to occur in the stomach. In pigs fed a standard ration hepatic portal venous VFA remained low for the first 4 h after feeding but then rose with the onset of large bowel fermentation. Feeding the diet supplemented with propionate caused hepatic portal venous plasma concentrations to rise by approximately 0.4 mumol/ml. This increase was apparent 30 min after feeding and was sustained for 3 h but subsequently there was no difference to controls. As in the rat, the absorption of dietary propionate appeared to occur in the upper gastrointestinal tract. The transport of propionate via the porcine hepatic portal vein also appeared insufficient to account for the dietary intake and suggests metabolism of the acid by the upper gastrointestinal tract. Further studies with perfused livers from fed rats indicated that propionate at a concentration of 1 mumol/ml did not alter cholesterol synthesis but that inhibition occurred at 18 mumol of propionate/ml. It appears that a redistribution of cholesterol from the plasma to the liver, rather than inhibition of hepatic and intestinal cholesterol synthesis, is responsible for the hypocholesterolaemic effects of dietary propionate. Because the absorption and transport of dietary propionate appears to follow a time course which differs considerably to that of the acid produced by the large bowel microflora, we conclude also that VFA produced by such fermentation would not seem to be responsible for the hypocholesterolaemic effects of certain water-soluble plant fibres.

A viscous fibre (methylcellulose) lowers blood glucose and plasma triacylglycerols and increases liver glycogen independently of volatile fatty acid production in the rat.

1. Adult male rats were maintained on diets containing 80 g methylcellulose/kg of low (25 cP), medium (400 cP) and high (1500 cP) viscosity. 2. After 10 d, the viscosity of stomach and caecal contents was found to have increased in proportion to that of the dietary fibre. Concentrations of volatile fatty acids in caecal digesta were lowest with the high-viscosity fibre but acetate was the major acid present with all three diets. Acetate was the only acid found in significant quantities in hepatic portal venous plasma and concentrations of this acid were unaffected by diet. 3. Concentrations of glucose in arterial blood were low with the medium- and high-viscosity diets while the content of liver glycogen was high. These effects of fibre were not directly on glucose absorption as the intestines were net removers of the hexose at the time of sampling. 4. Hepatic lipogenesis and plasma triacylglycerol concentrations were both higher in rats fed on the low-viscosity fibre. Plasma cholesterol concentrations, hepatic cholesterol synthesis and faecal bile acid excretion were not altered by dietary fibre viscosity. 5. We conclude that the effects of dietary fibre on carbohydrate absorption and storage and fatty acid synthesis are a function of the viscosity of the fibre in solution, high viscosity slowing the digestion and absorption of nutrients in the small intestine. Large-bowel microbial fermentation is not of direct significance to these events. In contrast, effects of fibre polysaccharides on sterol metabolism seem not to be related to their rheological properties.

Blood carbonmonoxyhaemoglobin levels are chronically elevated in alcoholics treated for detoxification.

Self-reported daily consumption of cigarettes and ethanol was recorded from a group of 59 ethanol-dependent persons admitted to a detoxification clinic at 2000-2400 h and averaged 40 cigarettes per day and 320 g ethanol per day, respectively. In consequence, concentrations of blood COHb and ethanol were high. Mean values for COHb were 8.5 and 9.9% in men and women, respectively, while the corresponding levels for ethanol were 220 and 280 mg/100 ml. Blood acetate concentrations were also elevated and correlated negatively with mixed venous oxygenhaemoglobin concentrations. The mean carbonmonoxyhaemoglobin concentration at 0700 h was found to be approximately 65% of that on admission. Biochemical and haematological analysis, past medical history and current physical examination revealed a high degree of ethanol and/or cigarette-related pathology. Studies on lipid and lipoprotein metabolism in ethanol-dependent persons should take into account their simultaneous high cigarette abuse.

Comparative effects of dietary wheat bran and its morphological components (aleurone and pericarp-seed coat) on volatile fatty acid concentrations in the rat.

Adult male rats were fed on diets containing 100 g dietary fibre/kg either as alpha-cellulose or wheat bran or the pericarp-seed coat or aleurone layers prepared from that bran by sequential milling and air elutriation and electrostatic separation. After 10 d, concentrations of total volatile fatty acids (VFA) in caecal fluid were significantly different between groups and fell in the order: aleurone greater than wheat bran greater than pericarp-seed coat greater than cellulose. This ranking probably reflected the ease of fermentation of fibre polysaccharides by colonic bacteria which also resulted in a considerably higher faecal bacterial mass in the aleurone group. Because of the differences in the volume of caecal digesta, the mass of caecal VFA was considerably the highest in the aleurone group, intermediate with wheat bran and equally low in the pericarp-seed coat and cellulose groups. The diet based on aleurone gave a relatively higher proportion of propionate but with both pericarp-seed coat and wheat bran the contribution of butyrate was raised. VFA concentrations in hepatic portal venous plasma were proportional to caecal concentrations with very high (greater than 3 mM) values being recorded in the aleurone group. The findings are discussed in relation to the apparent susceptibility of the morphological components of wheat bran to fermentation by large bowel bacteria.

Age-related augmentation of phosphorylase b kinase in hepatic tissue from the glycogen-storage-disease (gsd/gsd) rat.

The effects of food deprivation on body weight, liver weight, hepatic glycogen content, glycogenolytic enzymes and blood metabolites were compared in young and old phosphorylase b kinase-deficient (gsd/gsd) rats. Although the concentration of glycogen in liver from 9-week-old female gsd/gsd rats (730 mumol of glucose equivalents/g wet wt.) was increased by 7-8% during starvation, total hepatic glycogen was decreased by 12% after 24 h without food. In 12-month-old male gsd/gsd rats the concentration of liver glycogen (585 mumol of glucose equiv./g wet wt.) was decreased by 16% and total hepatic glycogen by nearly 40% after food deprivation for 24 h. Phosphorylase b kinase and phosphorylase a were present at approx. 10% of the control activities in 9-week-old gsd/gsd rats, but both enzyme activities were increased more than 3-fold in 12-month-old affected rodents. It is concluded that the age-related ability to mobilize hepatic glycogen appears to result from the augmentation of phosphorylase b kinase during maturation of the gsd/gsd rat.

Effects of food restriction and starvation-refeeding on volatile fatty acid concentrations in the rat.

Adult male rats were fed either ad libitum or at levels of 19 or 15 g of nonpurified diet per rat daily or subjected to 48 h of starvation followed by 24 h of refeeding. Concentrations of total and individual volatile fatty acids (VFA) in cecal contents were higher in rats fed ad libitum than in those restricted to 19 or 15 g/d. Only butyrate concentration was lower in rats given 15 g/d than in those given 19 g/d. In starved animals all cecal VFA declined within 24 h of food removal, but the greatest change was in butyrate, which fell to less than 12% of the initial value. Acetate and propionate fell further after 48 h, but their concentrations were restored to control values within 24 h of refeeding while butyrate remained depressed by 50%. Cecal pH was closely related to total VFA concentration, although the highest degree of correlation was with butyrate. Hepatic portal venous plasma VFA concentrations generally reflected those in cecal digesta except that the proportion of butyrate was relatively lower in this blood vessel than in cecal contents. Under all conditions acetate was the only VFA found in arterial plasma and in the fully fed state was lower than in hepatic portal venous plasma. Food restriction and starvation did not alter arterial concentrations, indicating abolition of net uptake. We conclude that all VFA are affected by availability of fermentable material to the large bowel microflora but that the disproportionate changes in butyrate may reflect preferential use of this acid by cells of the large bowel wall.

Time-course of changes in plasma lipids in diabetic rats fed diets high in fish or safflower oils.

Adult male rats were maintained for 10 days on a standard chow diet or that diet supplemented with either safflower or marine fish oils, and then rendered diabetic with streptozotocin (40 mg/kg of body weight) and circulating metabolites determined over the next 3 days. Pre-diabetic concentrations of glucose and insulin did not differ between groups, and the severity of hyperglycaemia and lowering of insulin in streptozotocin-treated animals were also similar. Pre-diabetic concentrations of plasma free fatty acids and triacylglycerols were lower, and blood ketone bodies were higher in non-diabetic rats fed fish oil than in both other groups. However, following streptozotocin treatment, plasma free fatty acids rose significantly more in both groups of oil-fed animals than in chow-fed ones. Plasma triacylglycerols were unaltered from pre-treatment levels in rats fed chow, but rose considerably in both groups fed oil-supplemented diets. In a subsequent experiment it was shown that the increase in triacylglycerols persisted for up to 11 days after streptozotocin and the hypertriglyceridaemia was greatest in the fish oil group. The rise would seem to result from defective clearance of lipoproteins of dietary origin. It appears that fish oil-supplemented diets should be avoided in diabetics until the possibility of increased hypertriglyceridaemia has been excluded by controlled studies.

Effects of wheat bran and porridge oats on hepatic portal venous volatile fatty acids in the pig.

Adult male pigs (40-60 kg of body weight) of the Kangaroo Island strain were surgically implanted with chronic indwelling hepatic portal venous cannulae. After a 24-hour fast the animals were given meals containing 500 g of either wheat bran or porridge oats and 200 g of sucrose and 2 litres of milk. With both cereal preparations plasma volatile fatty acids rose in the hepatic portal vein but the increase was significantly greater with wheat bran. Omission of sucrose and milk did not alter the response to porridge oats but diminished the response to wheat bran. These changes in plasma volatile fatty acids were unaffected by prior cooking of the cereals with hot water. With all test meals acetate and propionate were the major acids found, with butyrate contributing less than 8% of the total. This compositional profile was also found when the pigs were fed a commercial ration. The absence of butyrate differed from observations in the rat and reflected low concentrations of this acid in large bowel digesta. The difference in the response of the concentration of volatile fatty acids to feeding porridge oats and wheat bran in the pig was also the reverse of that found in the rat. These species differences may be of significance in relation to the choice of animal models for human fibre metabolism.

The adaptive effects of dietary fish and safflower oil on lipid and lipoprotein metabolism in perfused rat liver.

To elucidate the mechanisms underlying the plasma triacylglycerol-lowering effects of certain fish oils, livers from male rats fed either a standard commercial diet (controls) or diets supplemented with 15% (w/w) fish or safflower oils were perfused with undiluted rat blood. Rates of hepatic lipogenesis, measured by the incorporation of 3H2O into fatty acids, followed the order: control greater than safflower oil greater than fish oil. Secretion of newly synthesized fatty acids in very-low-density lipoproteins was also inhibited by the feeding of both oil-supplemented diets with the greater suppression being seen in livers from animals fed fish oil. The hepatic release of very-low-density lipoprotein triacylglycerol mass was also significantly depressed in animals fed the fish oil-supplemented diet but not in those fed safflower oil. Ketogenesis did not differ between livers from rats fed the control and safflower oil diets but was significantly raised in the fish oil group. Increased ketogenesis with fish oil was paralleled by a decrease in the sensitivity of carnitine palmitoyl transferase of isolated mitochondria to inhibition by malonyl-CoA. The inhibitory effect of malonyl-CoA in the safflower oil group was intermediate between that in the fish oil and control groups. Activities of glycerophosphate acyltransferase with either palmitoyl-CoA or oleyl-CoA were increased by feeding oil-supplemented diets. Activity with palmitoyl-CoA that was suppressible by N-ethylmaleimide was also considerably diminished in both groups. The results indicate that the lowering of plasma triacylglycerols by fish oil reflects: (a) diminished lipogenesis; (b) increased fatty acid oxidation possibly in peroxisomes; and (c) diminished secretion of triacylglycerols by the liver.

Isolation buffer calcium concentration and subsequent acute effects of insulin on metabolism of perfused rat liver.

During the surgical isolation for subsequent perfusion in situ with blood containing 1.3 mM Ca, livers from fed male rats were perfused with buffer containing 1.3 or 3.6 mM Ca. Total liver Ca was unaffected by buffer concentration and did not differ from non-perfused controls. In all groups a negative relationship was found between serum Ca and K+ concentrations and serum K+ was depressed by insulin and high buffer Ca while serum Ca was raised by the latter. Blood glucose was raised by isolation at high buffer Ca but was lowered by insulin with an increase in liver glycogen in both groups. Ketogenesis was unaffected by buffer Ca and was inhibited by insulin. The hormone raised serum triacylglycerol secretion and liver triacylglycerols in both groups, but the latter increase was abolished at the higher buffer Ca concentration. This effect on liver triacylglycerols may be related to the rise in microsomal Ca-ATPase activity found in these livers. We conclude that use of high concentrations of Ca in the isolation buffer, which may maintain hormone sensitivity in isolated hepatocytes, is not advantageous to the subsequent perfusion of the isolated liver with undiluted blood.