Physiological levels and action of dehydroepiandrosterone in Yucatan miniature swine.

The biological role of dehydroepiandrosterone (DHEA) and its less active sulphated conjugate DHEAS was investigated in two experiments using Yucatan miniature swine. In experiment 1, plasma levels of both DHEA(S) among males were greater than female pigs that ranged in age from 0.3 to 84 mo old (P < 0.0001). In males, DHEA(S) were related inversely to serum triglycerides; DHEA was positively related to triglycerides in females (P < 0.01). In experiment 2, four 2-yr old male pigs, used as their own control, showed a 5% decrease in body weight, 11% increase in energy expenditure, 88% increase in lipid, and 100% decrease in glucose utilization (P < 0.0001) in response to DHEA vs. placebo treatments when adjusted for body weight. Plasma DHEA(S) were not different between treatment conditions. Glucose tolerance and plasma insulin levels were not different from controls. In vivo response to norepinephrine indicated beta-adrenergic sensitivity was altered by DHEA. Present findings suggest DHEA and/or its hormone products are important in modulating energy expenditure and lipid utilization for energy in male animals. The role of DHEA in energy metabolism and the difference between sexes warrant further investigation.

Cyclic food restriction, insulin and mammary cell proliferation in the rat.

We reported recently that weight cycling significantly increased the incidence of mammary cancer in virgin female rats that were pretreated with N-methyl-N-nitrosourea. The present study investigated the effect of weight cycling on mammary epithelial cell proliferation and its relationship to changes in plasma insulin, estrogen, progesterone and urinary corticosterone in 30 female virgin Sprague-Dawley rats. Animals were fed a modified AIN-76A diet containing 24.6% corn oil by weight. Weight-cycled (WC) rats were food restricted daily by either 33% or 50% of non-restricted controls for 1 week followed by 3 weeks compensatory refeeding and weight recovery over 18 weeks or 4.5 weight cycles. WC rats consumed 6-10% less food than controls (P = 0.01) but showed a 71-89% greater efficiency of food utilization for growth (P < 0.0001) than controls. There were no differences in total weight gain during treatment. Mammary lobuloalveolar and ductal cell proliferation of WC rats, measured by 5-bromo-2'deoxyuridine labelling, increased in a dose-response fashion, P = 0.03, P = 0.06 respectively in comparison to controls. Energy and substrate utilization measured by indirect calorimetry indicated WC animals expended less energy (P = 0.005) and utilized less glucose (P = 0.0001) and protein (P = 0.006) during restriction, and less lipid during recovery (P = 0.05) than controls. There were no significant differences in hormone levels between groups. Multiple regression analysis with plasma insulin, estrogen, progesterone and urinary corticosterone as independent variables (r = 0.947, r2 = 0.897, P = 0.003) showed that plasma insulin was the only significant predictor (P < 0.01) of mammary cell proliferation. In accord with this observation, tyrosine-phosphorylated activation of insulin receptor substrate-1, detected by immunoprecipitation and Western immunoblot analysis in mammary tumors of WC rats from our previous study, was 3-5 times greater than in non-restricted controls (P < 0.01). Present findings suggest that weight cycling in rats increases risk of breast cancer development via insulin stimulated mammary cell proliferation.

Cyclic food restriction alters substrate utilization and abolishes protection from mammary carcinogenesis female rats.

We tested the hypothesis that cyclic food restriction abolishes protection against mammary carcinogenesis. Virgin female Sprague Dawley rats (n = 159) were injected intraperitoneally with 25 mg/kg n-methyl-n-nitrosourea at 50 d of age. Eleven days later, rats were given free access to a 24.6 g fat/100 g AIN-76A diet (ad lib-c), fed in two meals (me-c), or fed in two meals restricted in weight by 33% for 1 wk followed by 3 wk of compensatory refeeding (me-r) for 18 wk or 4.5 restriction cycles. Energy and substrate utilization of 15 rats from each group was measured by indirect calorimetry. The me-r rats ate and weighed less (P < 0.0001), had a greater efficiency of food utilization (P < 0.01), and had a 12% higher incidence of mammary cancer (P < 0.0001) than ad lib-c rats after adjusting for the effect of final body weight. Resting metabolic rate was not different among groups, but me-r rats used less glucose during restriction and more glucose and less lipid for energy during body weight recovery than me-c rats (P < 0.0001). Increased energy efficiency and the shift in utilization of glucose and fatty acids followed closely the effects of cyclic food restriction and meal feeding on mammary carcinogenesis.

Increased lipolysis to beta-adrenergic stimulation after dehydroepiandrosterone treatment in rats.

Young adult male rats were treated with 4 mg dehydroepiandrosterone (DHEA)/100-g diet for 4 wk or were fed the same purified diet unadulterated (51 carbohydrate:20 fat: 23.5 protein; wt/wt). After 1 wk body weight and fat mass of the DHEA-fed rats were significantly less than the controls. By the end of week 3, fat-free mass of the DHEA rats was less than the controls. Neither food intake nor resting metabolism, measured by indirect calorimetry, was different between groups. Isolated epididymal adipocytes of DHEA rats were significantly smaller and isoproterenol (x 10(7) M) stimulation of glycerol release was 53% greater (P < 0.01) than the controls. Basal rate of glycerol release increased significantly for both groups in response to the adenosine inhibitor adenosine deaminase; there were no significant interaction effects. Inhibition of lipolysis by the adenosine analogue phenylisopropyladenosine was similar between groups. Findings support the hypothesis that DHEA reduces adiposity directly by increased lipolysis, but the mechanism of action does not involve a change in the antilipolytic function of adenosine.

Dietary modulation of pancreatic carcinogenesis: calories and energy expenditure.

Physical activity (exercise) is a lifestyle factor that has received little attention with regard to its role in the etiology and/or prevention of cancer. These studies examined the effects of treadmill exercise on the early stages of pancreatic carcinogenesis initiated by azaserine in rats. Male Lewis rats were treated with azaserine at 2 weeks of age and weaned to experimental protocols at 3 weeks of age. Two experiments were undertaken; treadmill exercise began at 6 weeks of age (Experiment 1) or at 13 weeks of age (Experiment 2). Rats were exercised for 15-20 min/day and for 3-5 days/week. Treadmill speed and angle of incline were adjusted to afford a range of exercise intensities. The development of putative preneoplastic lesions of the pancreatic acinar cells (henceforth termed foci) was evaluated by quantitative stereological analysis using light microscopy. In Experiment 1, exercise resulted in a known paradoxical reduction in food intake by about 15% of the intake of the sedentary group fed ad libitum. The burden of azaserine-induced foci was decreased by approximately 37%, and this was attributed to the well known effects of reduced caloric intake in these young, rapidly growing rats. In Experiment 2, the higher intensity treadmill exercise group had an increased focal burden, compared to their sedentary pair-fed controls. Importantly, this enhancement occurred despite a reduction in food intake and body fat stores in this treadmill exercise group. These experiments demonstrate that exercise may suppress or promote carcinogenesis, depending upon the stage in the life cycle of the animal.

Effects of diet and exercise on the in vivo rates of the triglyceride-fatty acid cycle in adipose tissue and muscle of the rat.

The effects of feeding a low-protein diet (5 percent w/w) and daily exercise on the rates of substrate (futile) cycling between triglyceride and fatty acids (TG-FA cycle) were studied in rats in vivo using a radiochemical assay that involves following the incorporation of tritium from 3H-H2O into the fatty acid and glycerol moieties of triglyceride. Sixty-four rats were fed either a purified control diet (COND) consisting of 70 percent carbohydrate, 20 percent protein, 5 percent fat or an experimental low protein (LPD) diet consisting of 80 percent carbohydrate, 5 percent protein and 10 percent fat (w/w) and were either exercised six days weekly or remained sedentary for six weeks. Both LPD and exercise training (EXT) were found to increase significantly the rate of TG-FA substrate cycling above the rates observed in dietary and sedentary control groups. The LPD increased significantly the rate of cycling in interscapular brown adipose (IBAT), while adipose (WAT) and diaphragm muscle. EXT increased the rates of substrate cycling in soleus, heart, and diaphragm muscle and WAT. Rate of cycling in cardiac or skeletal muscle was one-twentieth that found in adipose tissue. There were also sex differences in the rate of substrate cycling. Substrate cycling in soleus and heart muscle of male animals were consistently higher than respective female treatment groups. Sedentary and EXT LPD animals weighed significantly less than but consumed a similar amount of food to the respective COND animals. These data provide the first in vivo evidence that the rate of substrate cycling can be increased by diet or by exercise training. The possible importance of TG-FA and other substrate cycles on efficiency of energy metabolism and etiology of obesity are discussed.

Fasting energy metabolism of the Yucatan miniature pig.

The fasting metabolic rates (FMR) of Yucatan miniature swine were determined using an open-circuit indirect respiration calorimeter. Mature nulliparous females had a mean FMR of 93 kcal/kg BW.75 and did not change significantly during the estrous cycle. This value is comparable to that observed in mature domestic swine. The calculated metabolizable energy requirement for maintenance for the Yucatan sow is 116 kcal/kg BW.75. Growing Yucatan boars had FMR of 127, 119 and 101 kcal/kg BW.75 at 15, 21 and 38 weeks of age, respectively, and were similar to values for comparably aged domestic swine. The calculated estimate for the metabolizable energy requirement for maintenance for Yucatan boars ranged from 158 kcal at 15 weeks of age to 126 kcal/kg BW.75 for 38 week old animals. Based on the similarity between the FMR of the Yucatan miniature swine in the present study and values published for standard size commercial hogs, it is concluded that the metabolic rates of these breeds of pig are similar. It is suggested that the daily energy needs of the Yucatan miniature pig may be met using values published for production livestock having similar physiological condition when adjusted for the smaller body size of the Yucatan breed.

Effect of type and amount of dietary fat on the enhancement of rat mammary tumorigenesis by exercise.

The effect(s) of treadmill exercise and type and amount of dietary fat on the process of mammary tumorigenesis was investigated. Female Sprague-Dawley rats were fed a purified 5% fat diet (AIN-76A) from 21 to 64 days of age. At 50 days of age each rat was intubated p.o. with 5 mg 7,12-dimethylbenz(a)anthracene (DMBA). Fourteen days after DMBA, the rats were randomized into one of three diet groups: 5% fat as corn oil, 24.6% fat as corn oil, or 24.6% fat as a mixture of palm (21.8%) and corn oil (2.8%). The combination of palm and corn oil provided the same amount of linoleic acid per g as the 5% corn oil diet. Half the animals receiving each diet were exercised on a treadmill at a speed of 20 m/min, 1 degree incline, 15 min/day, 5 days/week, and were designated as the moderate intensity treadmill exercise group (MITE). The remaining animals were exercised at a speed of 2 m/min, 1 degree incline, 15 min/day, 5 days/week, and were designated as the low intensity treadmill exercise group (LITE). The experiment was terminated 154 days after DMBA was administered. The median tumor-free time was significantly shortened in MITE rats receiving the 24.6% fat, corn oil-formulated diet in comparison to LITE rats receiving the same diet (43 day vs. 62 day, P = 0.028). Similarly, tumor appearance was more rapid in MITE rats consuming the low fat corn oil diet in comparison to the low fat diet-fed LITE group (57 day vs. 67 day, P = 0.046). Exercise exerted no effect on the rate of tumor appearance in rats that received the 24.6% palm and corn oil mixture, (58 day, MITE, vs. 62 day, LITE, P = 0.502). Mean body weight gains were similar among groups, although MITE rats consistently weighed more than LITE rats consuming the same diet. Gross carcass composition was unaffected by either the level of exercise or the amount of dietary fat consumed. The data indicate that moderate intensity treadmill exercise for a short duration, that is without effect on carcass fat content, can stimulate mammary tumorigenesis in rats fed low or high fat diets. This effect can be influenced by the type of dietary fat ingested.

Effect of exercise on the induction of mammary carcinogenesis.

Although data reported in several epidemiological investigations indicate that reduced consumption of dietary fat and increased levels of physical activity are associated with reduced risk for breast cancer, the results of some studies do not support these observations. Underlying this situation is the unanswered question about whether degree of body fatness, which is affected by dietary composition, total caloric intake, and energy expenditure, is the critical determinant affecting breast cancer risk. The objective of this work was to establish whether increasing energy expenditure by exercise would reduce the occurrence of mammary carcinomas induced by 7,12-dimethylbenz[a]anthracene (DMBA) in animals consuming a high fat diet to the level of occurrence observed in sedentary animals consuming a low fat diet. Female Sprague-Dawley rats were obtained at 21 days of age and maintained on a 5% (w/w) corn oil diet (AIN-76A) until they were 64 days of age. At 50 days of age, rats received either 5 mg DMBA or the solvent in which the carcinogen was dissolved. Fourteen days after DMBA intubation they were randomized into one of three groups: 5% fat (w/w), sedentary; 24.6% fat (w/w), sedentary; or 24.6% fat (w/w), exercised. Animals were exercised on a motor-driven treadmill at a belt speed of 20 m/min and a 1-degree incline for 15 min/day, 5 days/week for 18 weeks. Feeding a high fat versus a low fat diet increased the number of breast cancers induced and the rate at which they appeared in agreement with previous investigations. However, rather than retarding the development of tumors as was hypothesized, moderate treadmill exercise increased the incidence and number of cancers induced and shortened cancer latency in comparison to animals that received either the high fat or low fat diet and were sedentary. Body composition was not altered by the exercise regime imposed, although these animals weighed more than either sedentary group. These data document a heretofore unreported effect of a moderate level of aerobic work on breast cancer induction.

The effect of dehydroepiandrosterone acetate on liver peroxisomal enzyme activities of male and female rats.

The present study is a follow-up to the report that dehydroepiandrosterone (DHEA) acetate treatment in rats stimulated metabolic heat production and suppressed serum triglycerides, adiposity and weight gain without affecting food intake. Activities of peroxisomal fatty acyl-coenzyme (CoA) oxidase and catalase as well as mitochondrial citrate synthase were assayed in liver tissue of 24 young adult male and female Wistar rats fed a nonpurified diet containing 0.6% DHEA (6 g/kg) for 6 wk. DHEA-treated animals gained less weight but had heavier liver weights than did the controls. Hepatic activity of fatty acyl-CoA oxidase of the experimental male and female animal was 1058 and 946% higher, respectively, than that of the controls. For catalase activity, only the female groups were different (30%). Activity of citrate synthase was not affected by DHEA. These data support the hypothesis that the inhibitory effect of DHEA on energy storage as fat is mediated, at least in part, by increased beta-oxidation of fatty acids in peroxisomes. The peroxisomal beta-oxidation pathway is uncoupled from oxidative phosphorylation; electrons are transferred directly to molecular O2 because of cycling of NAD/NADH resulting in the expenditure of chemical energy as heat.

Effects of dehydroepiandrosterone acetate on metabolism, body weight and composition of male and female rats.

Twenty adult Sprague-Dawley outbred rats (10 male and 10 female) were fed a nonpurified diet without or containing dehydroepiandrosterone acetate (DHEA 6 g/kg diet) for 11 w. DHEA-treated animals weighed less than the controls after 6 wk and until the end of treatment. However, only the differences between male groups were statistically significant. Food intake of the DHEA-fed animals was not affected, but resting heat production was elevated for both sexes. Serum triglyceride levels and activity of hepatic glucose-6-phosphate dehydrogenase of the experimental groups were lower than controls. Analyses of body composition indicated DHEA-treated animals had proportionately less body fat and therefore more body water, protein and ash than controls. In most cases, differences in body composition were due primarily to effects of DHEA on the female animals. In a second experiment, DHEA treatment did not alter urinary ketone levels nor did it enhance citrate synthase activity in interscapular brown fat, skeletal muscle, heart or liver. Findings suggest that DHEA acetate treatment affected body weight, body composition and utilization of dietary energy by both impairing fat synthesis and promoting fat-free tissue deposition and resting heat production. Possible mechanisms by which DHEA may affect metabolism are discussed.

Effects of exercise-training on the thermic effect of food and body fatness of adult women.

Resting metabolic rate (RMR), thermic effect of food (TEF), aerobic capacity (VO2max), body fat, and food intake were measured in 10 healthy women before and after a 10 week graded exercise program of jogging. Pretraining TEF was a linear function of VO2max. Following exercise training, the women showed a significant increase (20%) in VO2max and loss (10.4%) of body fat; body weight did not change. Fat loss was directly related to changes in VO2max and RMR. The women showed a wide variation of changes in RMR (-21 to +2%) and TEF (-32 to +66%) from their pretraining levels. The changes in RMR and TEF were significantly positively correlated with improvements in VO2max. Analyses using multiple regression techniques, indicated that the changes in RMR and TEF accounted for 96.2% of the total variation in the changes of VO2max. Analyses of food intake indicated that diet composition (but not caloric intake) was highly related to the changes in VO2max, RMR, TEF and body fatness. Present results support further our hypothesis, that VO2max is an important physiological index of dietary thermogenesis and fat loss of individuals of normal body weight and fatness. Possible nutritional and physiological factors that may explain the wide variation in RMR and TEF of the women are discussed.

Effect of energy intake on the promotion of mammary carcinogenesis by dietary fat.

The effect of low-fat and high-fat diets on the induction of mammary carcinomas by 1-methyl-1-nitrosourea (MNU) was studied in female Sprague-Dawley rats. All rats were given MNU (25 mg/kg body wt) at 50 days of age. For the first 17 weeks after carcinogen administration, they were fed a purified diet containing either 5 or 20% fat incorporated into agar gel. Food intake was restricted, so that the amounts fed provided the same amount of net utilizable energy each day for both groups, regardless of the fat content of the diets. From 17 to 32 weeks, the diets were fed ad libitum. During the restricted feeding period, there was no significant difference in tumor incidence or in the number of tumors detected between the groups. During the weeks in which animals were fed ad libitum, significantly more tumors appeared in the high-fat group than in the low-fat group. The data provide support for the hypothesis that consumption of a high-fat diet can lead to an enhancement of mammary carcinogenesis. It appears, however, that diets must be consumed ad libitum for the stimulatory effect on tumor occurrence to be exhibited.

Dietary obesity and exercise in young rats.

Food intake, body weight, body composition, resting metabolic rate (RMR) and the thermic effect of food (TEF) were measured in young rats, some of which were fed a high energy (HE) diet and some of which were forced to swim daily. In general, high energy feeding as compared to chow feeding, resulted in higher food intake, higher body weight, higher body fat, and a slightly lower TEF. In many cases, however, the specific effects varied with the age and sex of the animals. Animals forced to swim weighed less; were leaner; and had higher RMR and TEF than sedentary animals. The effects of exercise on energy balance were greatest in males, while the effects of the high energy diet on energy balance were greatest in females. All HE-fed rats were switched to lab chow at 104 days of age. Body weights of sedentary HE-fed rats returned to control levels but those of exercised HE-fed rats did not. Both HE-fed groups remained fatter than chow-fed controls, even two months after the diet switch.

Effects of diet and exercise training on thermogenesis in adult female rats.

The effects of a cafeteria diet on body weight gain, food intake, resting metabolic rate (RMR) and the thermic effect of food (TEF) were compared in female Charles River albino rats that were either sedentary or exercise-trained. The food intakes of the exercise-trained rats on the cafeteria diet were increased to the same degree as those of the sedentary rats, however, they gained less body weight and body fat than sedentary controls. The exercise training increased RMR independent of diet, but differentially increased TEF in rats given the cafeteria diet. Conversely, sedentary rats on the cafeteria diet had significantly lower RMR, but their TEF were not different from control animals on lab chow. Thus, in addition to the direct cost of the exercise, training increased thermogenesis (RMR and TEF) which also helped prevent the dietary obesity which normally occurs with cafeteria diets.

Variations of dietary-induced thermogenesis and body fatness with aerobic capacity.

Resting metabolic rate (RMR), dietary thermogenesis (DIT), aerobic capacity (VO2 max), and percent body fat were measured in a group of sixteen women and ten men. DIT was significantly positively correlated with aerobic capacity (r = 0.658). Percent body fat was significantly negatively correlated with VO2 max (r = -0.727). RMR was independent of VO2 max, or body fatness, but women had a significantly higher average RMR than men. Three men and three women participated in an endurance training program which significantly increased VO2 max. DIT was increased in proportion to the increase in aerobic capacity (rs - 0.924), RMR was unchanged, and body fat decreased in males but not females. The controversy surrounding "luxuskonsumption" may stem in part from not taking into account individual differences in aerobic capacity when measuring DIT.

Overcompensation of food intake following brief periods of food restriction.

Four adult female Sprague-Dawley rats, maintained on an ad lib feeding schedule were deprived for either 0, 0.5, 1, 2, 4, 6, or 8 hours during the dark phase of the day-night cycle. It was found that the latency to initiate the first meal following the deprivation was independent of the previous deprivation interval. The animals were found to overcompensate for the periods without food by eating a large initial meal that increased proportionately in size with the duration of food restriction. Furthermore, the animals continued to overeat throughout the day. This deprivation-induced overeating by the animals resulted in an overcompensation in total food consumption that was 21-56% greater than on control days. The role of food intake as a regulator of body weight is discussed.

Effects of early cold adaptation on food efficiency and dietary-induced thermogenesis in the adult rat.

The food intakes of male and female rats (Harvard strain) adapted to cold (5 degrees C) starting at weaning were studied in the cold and during reacclimation to 25 degrees C. Following 24 hr food deprivation, the male rats regained the weight they had lost without making up for the food they had missed (increased food efficiency), while the female rats ate 75% more food than they had missed in regaining their body weights (decreased food efficiency), independent of the temperature at which the animals were housed. On a high fat diet, both cold groups gained less weight than did the two control groups. However, each female group gained significantly less weight than its same temperature male group. Dietary-induced thermogenesis was significantly enhanced in male cold adapted rats 3 months after reacclimation to the zone of thermal neutrality but not in cold adapted female rats. Sex and strain differences are discussed. These data indicate that early chronic cold exposure results in both immediate and long-term changes in food efficiency and metabolism in adult rats, but the effects vary with the sex of the animal.

Spillage behavior and thiamin deficiency in the rat.

Food spillage, food intake and body weight of four groups of adult rats were monitored daily as they were fed varying responses of deficient, marginal, or high thiamin (B) diet. Food spillage was a more sensitive index of the dietary B1 then either food intake or body weight change. Animals fed the deficient B1 diet exhibited the greatest levels of food spillage followed by the animals fed the marginal B1 diet. Hypophagia and weight loss occurred only among animals fed the marginal B1 diet. Feeding the high B1 diet to animals previously given the deficient and marginal B1 diets completely eliminated the excessive food spillage, and food intake and body weight returned to control levels. These data support the use of spillage behavior as a useful screening tool for detecting behavioral effects of inadequate nutrient intake or chemical intoxication.