Syed Q. Alam

Chemopreventive effects of beta-carotene and 13-cis-retinoic acid on salivary gland tumors.

The chemopreventive effects of beta-carotene and 13-cis-retinoic acid (RA) on chemically induced salivary gland tumors were studied in rats. Young male Sprague-Dawley rats were injected in one of the submandibular salivary glands with 1 mg of dimethylbenzanthracene (DMBA) dissolved in olive oil. The contralateral gland was injected with the vehicle alone. Rats were divided into four groups and were fed ad libitum a semisynthetic diet supplemented with 0 or 100 mg beta-carotene/kg diet, or 20 or 100 mg RA/kg diet. Rats were killed at 22 weeks after the DMBA treatment, and tumors were examined histologically. Tumors were generally found to be squamous cell carcinomas or poorly differentiated neoplasms resembling squamous cell carcinomas. The tumor incidence was slightly lower in rats fed the diet supplemented with beta-carotene. RA had no appreciable effect on tumor incidence. A high activity of gamma-glutamyl transpeptidase was histochemically demonstrated in the tumors. There were some mortalities in the beta-carotene and RA supplemented groups, especially in the group fed high levels of RA. This mortality appeared to be related to vitamin K becoming somewhat limited.

Effect of dietarytrans fatty acids on some membrane-associated enzymes and receptors in rat heart

Three groups of male weanling Sprague-Dawley rats were fed diets containing 20% corn oil, 20% partially hydrogenated soybean oil (PHSBO) or 18% PHSBO +2% corn oil. PHSBO contained about 48% of its total fatty acids astrans-octadecenoate. Rats were killed after 16–18 weeks of feeding the various diets, hearts were dissected and crude sarcolemma was prepared by differential centrifugation. The activities of ouabain-sensitive (Na++K+)ATPase were significantly lower in membranes of rats fed 20% PHSBO than the control rats fed 20% corn oil. The feeding of 2% corn oil with 18% PHSBO resulted in partial restoration of the enzyme activity. The maximum number of [3H]ouabain-binding sites (Bmax) was also lower in cardiac membranes of rats fed 20% PHSBO than those fed 20% corn oil. Similar to (Na++K+)ATPase activity, some restoration of the number of [3H]ouabain-binding sites was observed when 2% corn oil was fed with 18% PHSBO-containing diet. There was no difference in the binding affinity of the radioligand for the receptor among the 3 dietary groups. Adenylate cyclase activities (fluoride-, isoproterenol- and forskolinstimulated) were lower in membranes of rats fed 20% PHSBO or 18% PHSBO +2% corn oil than in the control group fed 20% corn oil. Density of the β-adrenergic receptor was the lowest in cardiac membranes of rats fed 20% PHSBO. The feeding of 2% corn oil with 18% PHSBO resulted in partial restoration of the maximum number of [3H]dihydroalprenolol (DHA)-binding sites. The affinity of the binding sites was, however, not affected by the type of the dietary fat. The results of this study suggest that dietarytrans fatty acids can affect the activities of (Na++K+)ATPase and adenylate cyclase and the density of digitalis and β-adrenergic receptors in rat heart.

(3H)Forskolin- and (3H)dihydroalprenolol-binding sites and adenylate cyclase activity in heart of rats fed diets containing different oils

The characteristics of the cardiac adenylate cyclase system were studied in rats fed diets containing fish oil (menhaden oil) and other oils. Adenylate cyclase activity generally was higher in cardiac homogenates and membranes of rats fed diet containing 10% menhaden oil than in the other oils. The increase in enzyme activity, especially in forskolin-stimulated activity, was associated with an increase in the concentration of the [3H]forskolin-binding sites in cardiac membranes of rats fed menhaden oil. The β-adrenergic receptor concentration was not significantly altered although the affinity for [3H]dihydroalprenolol-binding was lower in membranes of rats fed menhaden oil than those fed the other oils. ω-3 fatty acids from menhaden oil were incorporated into the cardiac membrane phospholipids. The results suggest that the observed increase in myocardial adenylate cyclase activity of rats fed menhaden oil may be due to an increase in the number of the catalytic subunits of the enzyme or due to a greater availability of the forskolin-binding sites.

Lung surfactant and fatty acid composition of lung tissue and lavage of rats fed diets containing different lipids

Two nutritional models, essential fatty acid (EFA) deficiency and the feeding of saturated vs unsaturated fats, were used to determine the effects of dietary lipids on the fatty acid composition of rat lung and lavage. Semipurified diets containing 7% corn oil, 7% hydrogenated coconut oil (EFA-deficient), 10% butter or 10% safflower oil were fed to dams during lactation and thereafter to their offspring for a total of 24 weeks. Lipids were extracted from the lung lavage and lung tissue and their fatty acid composition was determined. The content of dipalmitoylphosphatidylcholine (DPPC), the main surfactant in the lungs, was also determined. The results show that the levels of DPPC in the lungs of rats fed 10% butter decreased although the decrease in the EFA-deficient rats was greater. Comparing rats fed butter with those fed corn oil, there were also modifications in the fatty acid composition of the total lipids and phospholipids of lung tissue and lavage as well as in phosphatidylcholine, phosphatidylethanolamine and phosphatidylinositol +phosphatidylserine fractions isolated from the lung tissue. The changes in fatty acid composition were somewhat fewer in rats fed butter then in those fed an EFA-deficient diet. The results suggest that a marginal EFA deficiency produced in rats by long-term feeding of 10% butter may account for the reduction in DPPC levels and in the changes in fatty acid composition in the lung tissue and lavage.

Incorporation oftrans fatty acids into submandibular salivary gland lipids

Three groups of rats were fed diets containing 20% corn oil, 20% margarine stock (MS) or 19% MS +1% corn oil. Diets were fed for 12 weeks, 1 week of pregnancy, 3 weeks of lactation and 8 weeks post-weaning. The incorporation oftrans-octadecenoate into various lipids of the submandibular salivary gland (SMSG) homogenates and plasma membranes was studied.Trans octadecenoate was incorporated into all the lipid fractions studied. Its levels were the highest in phosphatidylethanolamine. The double bond index of phospholipid fatty acids in the plasma membranes of the SMSG was substantially lower in the group fed 20% MS. The fluorescence polarization of 1,6-diphenyl-1,3,5-hexatriene (DPH) was generally higher in the membranes of SMSG from rats fed MS than that of the other two groups, thus indicating lower fluidity. Also, the breakpoints in fluorescence polarization were at a higher temperature in the membranes from rats fed MS as compared with those fed corn oil. Lower fluidity of plasma membranes of SMSG observed in rats fed 20% MS may result in modification of the activities of membrane-bound enzymes.

Phospholipid composition of liver in rats fed high levels of 13-cis retinoic acid

The composition of liver phospholipids was studied in rats fed for 4 weeks diets containing 0, 100 or 300 mg 13-cis retinoic acid per kg diet. There was a significant decrease in phosphatidylcholine content, whereas the levels of phosphatidylethanolamine were slightly increased in liver phospholipids of rats fed 13-cis retinoic acid. The fatty acid composition of total phospholipids, PC, PE, and PI+PS fractions revealed a general increase in the levels of 18∶2 and 20∶3ω6, whereas the levels of 20∶4ω6 and C22 fatty acids were reduced in most of the hepatic phospholipids isolated from rats fed 13-cis retinoic acid containing diets. A decrease in the double-bond index of fatty acids was also observed in phospholipids of rats fed 13-cis retinoic acid. The data suggest that high levels of 13-cis retinoic acid may possibly be influencing the activities of microsomal desaturating and chain-elongating enzymes in the liver.

Reversal of diet-induced changes in adenylate cyclase activity and fatty acid composition of rat submandibular salivary gland lipids

This study sought to determine if the diet-induced changes in submandibular salivary glands can be reversed. Two groups of male weanling Sprague-Dawley rats were fed purified diets containing 9% butter + 1% corn oil (group I, control) or 9% ethyl ester concentrate of n-3 fatty acids + 1% corn oil (group II, experimental). After 5 weeks of feeding the respective diets, rats in group I were divided into two subgroups: Ia, which was maintained on the control diet, and Ib, which was shifted to the experimental diet for the reversal study. The rats in the experimental group were kept on their original diet. After five further weeks of feeding, the rats were killed, and membranes from submandibular glands were prepared and assayed for adenylate cyclase activity and for the fatty acid composition of total phospholipids. Changes characteristic of feeding n-3 fatty acids, including a significant increase in membrane fluidity as measured by the degree of unsaturation of fatty acids, were observed in the total phospholipids of membranes from the experimental group. The adenylate cyclase activity was two- to three-fold higher in membranes of rats fed the experimental diet (group II) than the control diet (group Ia). Whereas the diet-induced changes in fatty acid composition and membrane fluidity were largely reversed (group Ib, reversal study), changes in adenylate cyclase activity were only partially reversed. The results suggest that, in addition to the fatty acid composition and membrane fluidity, other factors may also be important in modifying adenylate cyclase activity.

Adenylate cyclase and β-receptors in salivary glands of rats fed diets containingtrans fatty acids

The effects oftrans fatty acids on adenylate cyclase were determined in the submandibular salivary glands (SMSG) of rats fed diets containing 20% corn oil, 20% partially hydrogenated soybean oil (PHSBO) or 18% PHSBO + 2% corn oil. The fluoride-and isoproterenol-stimulated adenylate cyclase activities were higher in the SMSG from rats fed 20% PHSBO than in the control group fed 20% corn oil. The feeding of 2% corn oil with the diet containing 18% PHSBO resulted in a complete restoration of isoproterenol-stimulated and a partial restoration of fluoride-stimulated adenylate cyclase activity. There was no significant difference in the concentration of the β-adrenergic receptor or in the receptor-binding affinity constants among the three dietary groups as measured by [3H]dihydroalprenolol (DHA)-binding. Higher fluorescence polarization of diphenyl hexatriene (DPH) was observed in SMSG membranes of rats fed diet containing 20% PHSBO than in the other two oils, suggesting that membrane fluidity may play a role in adenylate cyclase activity.

Studies on lacrimal gland lipids in essential fatty acid deficiency.

Summary Two groups of young, male, weanling rats were fed purified diets, one containing 7% corn oil (control) and the other 0% fat (EFA-deficient). After sixteen weeks of feeding, rats were killed, their extraorbital lacrimal glands were extracted for lipids, and the lipid composition was studied. Total phospholipid concentration was higher whereas total cholesterol, free cholesterol and cholesterol to phospholipid ratios were the same in lacrimal glands of EFA-deficient rats as compared to the controls. Total fatty acid concentrations were the same in the two groups. However, free fatty acid levels were reduced in the EFA-deficient gland. The fatty acid composition of total lipids, free fatty acids, triglycerides, phosphatidylcholine, phosphatidylethanolamine, and phosphatidylserine + phosphatidylinositol fractions obtained from the lacrimal glands of EFA-deficient rats showed changes in their fatty acid patterns which are characteristic of EFA deficiency, i.e., an increase in 16:1 and 18:1, a decrease in 18:2 and 20:4 and an accumulation of 20:3 ω9 fatty acids.

Pulmonary lipid peroxides and fatty acids of rats fed different lipids and exposed to oxygen at hyperbaric pressure

Semipurified diets containing different lipids were fed to rat dams during lactation and subsequently to their pups for 33 weeks post-weaning. Some rats within each group were exposed to oxygen at hyperbaric pressure (OHP). Lipid peroxide levels were lower in lungs of rats fed 7% hydrogenated coconut oil or 10% butter as compared with their controls, fed 7% corn oil or 10% safflower oil, respectively. Exposure to OHP increased lung peroxide levels. This increase varied with the type of fat in the diet. Studies of the fatty acid composition indicate that lipid peroxide levels generally increased with an increase in the levels of 18∶2 in lung total lipids. The results suggest that the type of dietary lipid may alter the susceptibility of the animal to pulmonary oxygen toxicity.