Tissue culture of cocoa beans (Theobroma cacao L.): Incorporation of acetate and laurate into lipids of cultured cells

Suspension cultures of cocoa bean tissue readily incorporated exogenous acetate into lipids. The distribution of radioactivity from acetate in individual lipid classes after 48 hr was 20, 5, 1, 15, 25, and 35% in triglycerides, diglycerides, free fatty acids, sterol esters, sterols and polar lipids, respectively. The labeled acetate was rapidly incorporated into various fatty acids within 2 hr. The [1-¹⁴C] saturated fatty acids declined slightly after 4 hr, whereas [1-¹⁴C] oleate declined significantly after 2 hr. There was a concomitant increase in [1-¹⁴C] linoleate. The radioactivity associated with linolenate was relatively high up to 4 hr, declined by 24 hr, and then increased again. The kinetics of fatty acid labeling suggested that biosynthesis of linolenic acid in cocoa bean suspension culture may occur via the desaturation of linoleic acid and the chain elongation of dodecatrienoic acid. The patterns of fatty acid radiolabeling following incubation of cells with [1-¹⁴C] laurate was consistent with this mechanism.     

Effect of nutritional status on the fatty acid composition of rat liver and cultured hepatocytes

The lipid concentration and fatty acid composition of the whole liver and of cultured hepatocytes isolated from the livers of rats fed ad libitum (fed), fasted for 24 hr (fasted), or fasted for 48 hr and then refed a fat-free, high carbohydrate diet for 48 hr (refed) was studied. Hepatocytes were maintained as monolayer cultures in serum-free, lipid-free media and their fatty acid composition was analyzed at 3, 24, 48, 72 and 96 hr. The livers of fed animals, as well as their hepatocytes, contained less total lipid than those from animals on either of the other dietary regimes. Livers of fasted animals had three times the amount of lipid found in the livers of fed animals, and the livers of refed animals contained five times the amount of lipid as the livers of fed animals (all based on mg lipid/g wet weight of liver). The fatty acid composition of hepatocytes after 3 hr of culturing was very similar to that of fresh liver when compared in each of the dietary regimes. However, while the fatty acid compositions of livers and hepatocytes from fed and fasted animals were similar, the pattern in liver of refed animals was quite distinct from that of the fed animals. In the fed and fasted animals palmitic acid (16∶0), stearic acid (18∶0), oleic acid (18∶1[n-9]), linoleic acid (18∶2[n-6]) and arachidonic acid (20∶4[n-6]) were the major fatty acids of the liver; in refed animals 16∶0, palmitoleic acid (16∶1[n-7]), 18∶0, 18∶1(n-9) andcis-vaccenic acid (the n-7 isomer of oleic acid) were the major fatty acids. During maintenance in culture the 18∶1(n-9) content of the hepatocytes increased in cells from livers of animals on all three dietary regimes. The polyunsaturated fatty acid content was similar in fresh livers and isolated hepatocytes in all samples when compared on the basis of μg fatty acid/mg of hepatocyte or liver protein. It was also found that the polyunsaturated fatty acid content of hepatocytes was remarkedly stable with time of culture when the cells were incubated in serum-free, lipid-free medium. Thus, isolated hepatocytes maintained in serum-free medium appear to be a possible system for the evaluation of the effects of prior nutritional status on fatty acid metabolism in the whole animal, not subject to hormonal and other somatic influences which often complicate the interpretation of such nutritional studies.     

Lipids of cultured hepatoma cells: III. Triglyceride and phosphoglyceride biosynthesis in minimal deviation hepatoma 7288C

Minimal deviation hepatoma 7288 C cells were cultured on media containing 25% serum to the confluent stage. The growth media was replaced with serumfree media containing 1-¹⁴C-palmitate, and incubations were continued for 0.75, 1.5, 3, 6, 12, and 24 hr. The distribution of radioactivity among the major neutral lipids and phosphoglycerides was determined for cells and culture media. Radioactivity in individual fatty acids of cellular triglyceride, phosphatidylcholine, and phosphatidylethanolamine also was determined. After 24 hr, more than 95% of the administered radioactivity was recovered in neutral and phosphoglycerides, indicating that only a small amount of the fatty acid was oxidized. At any time period examined, over 80% of the incorporated radioactivity was found in triglyceride, phosphatidylcholine, and phosphatidylethanolamine. Incorporation of the label into cellular triglyceride and phosphatidylcholine plateaued at 12 hr, whereas incorporation of radioactivity into phosphatidylethanolamine still was increasing at 24 hr. In contrast, during the entire incubation period the relative distribution of¹⁴C among esterified lipid classes in the culture media remained constant. Elongation of palmitic acid to stearic acid and its subsequent desaturation to oleic acid suggests that these cells possess an active elongation and monoenoic desaturation system. Labeled glycerol ether diesters were not detected in the cells or culture media. Positional distribution of the¹⁴C label in the triglyceride and phosphatidylcholine suggests that minimal deviation hepatoma cells do not exhibit diglyceride selectivity in the biosynthesis of these two lipid classes.     

The absorption of fatty acids by functional bovine mammary cells

Freshly dispersed bovine mammary cells rapidly absorbed long chain fatty acids from the culture medium. Differences in the rates of absorption were observed, i.e., palmitic > stearic > oleic > myristic > linoleic acid. The preponderance of the fatty acids absorbed were esterified into triglycerides (>75%) and the remainder were mostly incorporated into phospholipids. The cells secreted triglycerides into the culture medium. Of the phospholipid classes, phosphatidylcholine always contained most of the radioactivity in all experiments with labeled fatty acids. These observations are related to the metabolism of mammary cells in vivo.     

The influence of dietary fat on the lipogenic activity and fatty acid composition of rat white adipose tissue

The in vivo fatty acid synthesis rate, selected enzyme activities and fatty acid composition of rat white adipose tissue from animals fed semisynthetic diets of differing fat type and content were studied. All animals were starved for 48 hr and then refed a fat-free (FF) diet for 48 hr. They were then divided into three groups. One group was continued on the FF diet for 48 hr. Another group was fed a diet containing 44% of calories from corn oil (CO). The final group was fed a diet containing 44% of calories from completely hydrogenated soybean oil (HSO). The animals on the FF diet had a marked increase in adipose tissue fatty acid synthesis during the 96-hr feeding peroid (as measured by³H incorporation into adipose fatty acids). Addition of either CO or HSO to the diets did not significantly inhibit fatty acid synthesis in dorsal or epididymal adipose tissue. The activities of the enzymes' fatty acid synthetase, ATP-citrate lyase and glucose-6-phosphate dehydrogenase increased on the FF diet and generally were not inhibited significantly by the addition of either fat to the diets. Linoleic acid was the major polyunsaturated fatty acid (ca. 22%) in adipose tissue. Monounsaturated fatty acids (palmitoleic, oleic,cis-vaccenic) made up ca 38% of the total adipose fatty acids, while saturated fatty acids accounted for about 32% (myristic, palmitic and stearic). White adipose tissue in mature male rats was a major depot for n−3 fatty acids. There were differences in the fatty acid composition of epididymal and dorsal adipose tissue, particularly in their content of long chain, polyunsaturated fatty acids with epididymal tissue containing more of these compounds than dorsal fat. The fatty acid composition of the white adipose tissue did not change significantly during fasting or 96 hr of refeeding the FF diets. The addition of HSO to the diet for 48 hr had little influence on the adipose tissue fatty acid composition, but the addition of CO to the diet caused a 7% increase in the dorsal adipose tissue linoleate content (as percentage of total dorsal adipose tissue fatty acids) within 48 hr compared to animals fed the stock diet and those starved for 48 hr. The fatty acid synthesis data indicated that adipose tissue in the rat can continue to be a source of de novo fatty acid synthesis in animals consuming high-fat diets.     

Formation of diglycerides of long turnover time from labeled acetate and glucose in rat tissues

Rat adipose tissue pieces were incubated with acetate-2-¹⁴C and glucose-¹⁴C(U), respectively, and liver slices with acetate-2-¹⁴C. The labeled tissues were then reincubated in inactive medium, and the changes of radioactivity in the different lipid classes were determined. In all three experiments a significant amount of radioactivity was incorporated in the diglycerides. During 1 hr of reincubation in inactive medium the radioactivity of diglycerides decreased from 35 to 26% of the total lipid activity in the adipose tissue labeled with acetate. In the adipose tissue labeled with glucose radioactivity fell from 25 to 19%. In liver slices 11% of the labeled acetate was incorporated in the diglycerides, and during the 2 hr of reincubation this value fell to its half. The radioactivity of the uniformly labeled glucose was distributed equally in the fatty acids and the glycerol. The distribution of radioactive glycerol between diglycerides and triglycerides was similar to that of the labeled fatty acids. Triglyceride synthesis seems to always be accompanied by the formation of diglycerides with a lastint turnover time.     

Uptake and utilization of 1-14C palmitic acid by heart cells treated with fresh or thermally oxidized fats

The effects of fractions isolated from thermally oxidized corn oil or olive oil on the metabolic activity of heart endothelial and muscle cells were studied. Rat heart cells in culture, exposed to thermally oxidized fat components, took up more exogenous 1-¹⁴C-palmitic acid and incorporated more of it into the cell triacylglycerol fraction than when the cells were treated with fresh fats. Particularly with the heated corn oil compared to fresh corn oil, much less of the radioactivity from the labeled palmitic acid was deposited in the phospholipid fraction. Also, with heated corn oil when the incubation period was extended beyond 12 hr, there was a decline in the radioactivity retained in the triacylglycerol fraction of the heart muscle cells. When the fresh fats were compared for¹⁴C-radioactivity incorporation into the heart cells, the olive oil gave much higher values, indicating a distinct difference in response to the proportion of fatty acids supplied. Presented in part at the AOCS Meeting, Chicago, September 1976.     

The Influence of High Contents of Animal and Vegetable Fat in Concentrates on Rumen Fermentation Pattern and Milk Fatty Acid Composition in Dairy Cows

In two feedings trials with respectively fresh cows and cows in mid-lactation the animals received concentrates which provided ca. 400 g and 370 g fat from coconut and palmkernel expeller and palmkernels per day. The secretion of medium chain fatty acids with the milk was decreased as compared to cows on a control treatment. In the trial with cows in mid-lactation this decrease was compensated by a higher supply of the mammary tissue with lauric and myristic acid via the blood. The primary cause of the decreased secretion of medium chain fatty acids seems to be a disturbance of the rumen fermentation. A daily quantity of 345 or 620 g soyabean oil in ca. 13 kg concentrate also diminished the secretion of medium chain fatty acids in fresh cows. The negative effect of soyabean oil was probably partially caused by inhibition of the de novo synthesis of fatty acids in mammary tissue by long chain fatty acids, especially trans-isomers of C18-fatty acids. However, there were also indications of disturbance of rumen fermentation the more as the soyabean oil was mixed up the the free form as compared to soyabean oil in toasted soyabeans. Renderers fat and calcium salts of palmitic and oleic acid had a less negative effect on rumen fermentation. Still the secretion of medium chain fatty acids with the milk was decreased. This could be caused by inhibition of the de novo synthesis. The lowered production of medium chain fatty acids was compensated for or even exceeded by the extra supply of long chain fatty acids by the blood.     

Metabolism of 1-14C linolenic acid in developing brain: II. Incorporation of radioactivity from 1-14C linolenate into brain lipids

Metabolism of 1-¹⁴C linolenic acid was studied in growing animals by injecting the tracer intraperitoneally into 12–13 day old suckling rats and following up the results by sacrificing groups of animals at 8 hr, 48 hr, 15 day, and 45 day intervals. In the first 15 days, there was a greater decrease in radioactivity of brain total lipids compared to the later period, although the earlier age period is characterized by lipid deposition rather than breakdown. Since the 18∶3 ω3 family of fatty acids occurs largely in the brain total phosphatidyl ethanolamine fraction, we expected that, in the initial period, total phosphatidyl ethanolamine would be the most highly radioactive component. However, results showed that 8 hr after the tracer phosphatidyl choline had the highest specific radioactivity. When the total phosphatidyl ethanolamine fraction was resolved into diacyl and alk-1-enyl species, it was found that radioactivity was not distributed evenly between the two species. There was a progressive increase in radioactivity of the alkenyl and a decrease in the diacyl species. Forty-eight hr after the tracer, however, the radioactivity of phosphatidyl ethanolamine increased and at 45 days remained slightly higher than phosphatidyl choline. Radioactivity of cholesterol, a result of synthesis from acetate undoubtedly derived from the breakdown of tracer linolenate, was also high 48 hr after tracer and remained high until 45 days.     

Fatty acid specificity of glyceride synthesis by homogenates of bovine mammary tissue

Fatty acid esterification by cell free preparations of bovine mammary tissue was investigated to determine if the type of long chain fatty acid supplied might influence the rate of triglyceride synthesis by that tissue. Homogenates of lactating bovine mammary tissue esterified¹⁴C-fatty acids into glycerides at rates dependent upon chain length and degree of unsaturation. Palmitic, stearic, oleic and linoleic acids were esterified at rates consistent with their concentration in milk fat. A comparison of free fatty acid concentrations of mammary tissue with levels saturating esterification suggested that supply of fatty acids does not limit glyceride synthesis. Certain combinations of fatty acids were facilitory, competitive or inhibitory to esterification. Stearic acid complimented esterification of palmitic and oleic acids. Unlabeledtrans-11-octadecenoic acid did not compete with¹⁴C-palmitate as efficiently in the esterification process as did unlabeledcis-9-octadecenoic acid, indicating that the mammary gland may preferentially esterify thecis-isomer of C-18∶1. Linoleic acid inhibited esterification of palmitic, stearic and oleic acids. Michigan Agricultural Experiment Station Journal Article No. 5100.     

Bioactivity and emerging role of short and medium chain fatty acids

Cardiovascular disease (CVD) and insulin resistance are directly linked to overweight and obesity. Thus, any dietary strategy capable of causing weight reduction will lower CVD and diabetes risk. Oils rich in medium‐chain saturated fatty acids (MCFA) are among several dietary components that may have potential in the treatment of obesity. MCFA are less energy dense and highly ketogenic compared to long‐chain saturated and unsaturated fatty acids (LCFA). MCFA also differ from LCFA in their digestive and metabolic pathways, since they are easily oxidized and utilized as energy, with little tendency to deposit as body fat. The dietary intake of short (SCFA) and medium‐chain saturated fatty acids from natural food sources is approximately 2.4 g/day and accounts for about 9% of the total saturated fatty acid (SFA) intake. Although early clinical studies with high levels of MCFA resulted in increased levels of plasma triacylglycerols (TAG) and low‐density lipoprotein cholesterol (LDL‐C), and reduced levels of high‐density lipoprotein cholesterol (HDL‐C) compared to diets enriched in unsaturated LCFA, these adverse effects have not been observed in more recent studies with smaller more realistic amounts of MCFA. The lower caloric value of SCFA and MCFA and their unique metabolic features form the basis for their clinical use in enteral and parenteral nutrition and for novel reduced calorie lipids for use in conventional food products.     

Uptake and release of fatty acids by rat adipose tissue: Last in—First out?

Fatty acid labeled chyle was administered iv to ad lib fed rats. At intervals from 1 hr to 50 days later rats were killed and pieces of their parametrial adipose tissue were incubated in vitro with norepinephrine. The specific radioactivity of the mobilized free fatty acids was compared to that of tissue glycerides and that of tissue free fatty acids. The results indicate that fatty acids taken up by the adipose tissue do not mix immediately with the bulk of tissue lipids and that the mobilized free fatty acids do not pass through the tissue free fatty acid pool.     

Quality of subcutaneous,intermuscular and intramuscular fat tissue using elevated quantities of medium chain fatty acids in pig fattening

The present study comprised fat tissue samples of 46 (partly 23) pig carcasses randomly obtained from each one of four production systems: common fattening, pigs fattened on a low-fat diet and pigs grown on diets enriched with medium chain fatty acids (MCFA) in low or high amounts (0.3% and 3.6% C₈ with C₁₄, respectively). As models for subcutaneous, intermuscular and intramuscular fat tissues, back fat, dissected belly fat tissue and belly meat were used. In all fat tissues, the contents of MCFA were significantly elevated only with the high dietary content of MCFA, with a preferential retention of the MCFA in the storage tissues. In the MCFA supplemented groups, linoleic acid contents were slightly lower in subcutaneous and intermuscular fat as compared to the control group, in the group with the low-fat diet linoleic acid was considerably lower in all tissues. In spite of the only marginal differences in fatty acid pattern, the penetrometrically assessed firmness of backfat as well as the oxidative resistance of back and belly fat were almost twice as high in the high-MCFA group as in the other groups. In the low-fat group, water content of the back fat (16.9%) was higher than the average of the other groups (14.5%). The implications for routing assessment of fat tissue properties at slaughter plants are discussed.     

Hormonal response to lipid and carbohydrate meals during the acute postprandial period

Background

Optimizing the hormonal environment during the postprandial period in favor of increased anabolism is of interest to many active individuals. Data are conflicting regarding the acute hormonal response to high fat and high carbohydrate feedings. Moreover, to our knowledge, no studies have compared the acute hormonal response to ingestion of lipid and carbohydrate meals of different size.

Methods

We compared the hormonal response to lipid and carbohydrate meals of different caloric content during the acute postprandial period. Nine healthy men (22 ± 2 years) consumed in a random order, cross-over design one of four meals/beverages during the morning hours in a rested and fasted state: dextrose at 75 g (300 kcals), dextrose at 150 g (600 kcals), lipid at 33 g (300 kcals), lipid at 66 g (600 kcals). Blood samples were collected Pre meal, and at 0.5 hr, 1 hr, 2 hr, and 3 hr post meal. Samples were assayed for testosterone, cortisol, and insulin using ELISA techniques. Area under the curve (AUC) was calculated for each variable, and a 4 × 5 ANOVA was used to further analyze data.

Results

A meal × time effect (p = 0.0003) was noted for insulin, with values highest for the dextrose meals at the 0.5 hr and 1 hr times, and relatively unaffected by the lipid meals. No interaction (p = 0.98) or meal (p = 0.39) effect was noted for testosterone, nor was an interaction (p = 0.99) or meal (p = 0.65) effect noted for cortisol. However, a time effect was noted for both testosterone (p = 0.04) and cortisol (p < 0.0001), with values decreasing during the postprandial period. An AUC effect was noted for insulin (p = 0.001), with values higher for the dextrose meals compared to the lipid meals (p < 0.05). No AUC effect was noted for testosterone (p = 0.85) or cortisol (p = 0.84).

Conclusions

These data indicate that 1) little difference is noted in serum testosterone or cortisol during the acute postprandial period when healthy men consume lipid and dextrose meals of different size; 2) Both testosterone and cortisol experience a drop during the acute postprandial period, which is similar to what is expected based on the normal diurnal variation--feeding with lipid or dextrose meals does not appear to alter this pattern; 3) dextrose meals of either 75 g or 150 g result in a significant increase in serum insulin, in particular at 0.5 hr and 1 hr post-ingestion; 4) lipid meals have little impact on serum insulin.     

Lipid composition and erucic acid in rat liver cells in culture

Erucic acid (Δ¹³-docosenoic acid) was added to fetal calf serum, then fed to rat liver epithelial cells in culture, and uptake measured at intervals over 24 hr. During the first 6 hr. of incubation, uptake of the docosenoic acid was 21 nmoles/hr/mg protein in 7-day cells, and 15 mmoles/hr/mg protein in 14-day cells. Of¹⁴C-labeled erucic acid taken up by the cells in 24 hr, radioactivity measurements showed 60% of the total lipid¹⁴C activity derived from [1-¹⁴C] 22∶1 in neutral lipid (NL) and 40% in phospholipid (PL); whereas 55% of lipid¹⁴C activity was in NL and 45% in PL when the substrate was [14-¹⁴C] 22∶1. Within the NL fraction, 75% of¹⁴C activity derived from [1-¹⁴C] 22∶1 was in triglyceride (TG) and 11% in cholesterol (CHL), while 79% was in TG and 6.5% in CHL when the substrate was [14-¹⁴C] 22∶1. Triglycerides and cholesteryl esters accumulated in the cells during incubation with erucic acid. Among phospholipids separated by thin layer chromatography, 75% of¹⁴C activity was in lecithin (PC), 10% in phosphatidylethanolamine (PE), 5% in sphingomyelin (SPH), and 1% or less in cardiolipin (DPG). The highest specific activity (SA) was in PC, followed by SPH and PE. Incubation with erucic acid altered fatty acid composition of PC, PE, and SPH, although amounts of phospholipids were unaffected. Gas liquid chromatography analyses detected 18% erucic acid in PC, 2% in PE, and 4–5% in SPH.     

Effect of specific dietary fatty acids on lipogenesis in the livers and mammary glands of lactating mice

The effects of linoleic, linolenic and columbinic acids fed as 4% of a high carbohydrate (50% glucose) diet on the activities and the amounts of several enzymes associated with fatty acid synthesis in livers and mammary glands of lactating mice were compared with those for stearic and oleic acids. Fatty acid synthesis, measured in vivo, was significantly lower in livers of mice ingesting all 3 polyunsaturated fatty acids (PUFA), whereas in mammary glands synthesis was lower only in mice receiving columbinic acid. The activities of fatty acid synthetase (FAS) and acetyl CoA carboxylase were significantly reduced in liver by all 3 PUFA, as were activities of glucose-6-phosphate dehydrogenase, malic enzyme (ME) and citrate cleavage enzyme (CCE), also associated with lipogenesis. In mammary gland, on the other hand, the activities of these enzymes were unaffected by dietary PUFA. The tissue contents of FAS, ME and CCE, measured by rocket immunoelectrophoresis, were found to be significantly reduced in liver by linoleate, linolenate and columbinate but were not significantly altered in mammary gland. The decrease in hepatic lipogenesis observed was principally due to a decrease in the amounts of these enzymes induced by the dietary PUFA but the inhibition in mammary gland caused by columbinate could not be accounted for by a reduction in enzyme contents and therefore may be due to allosteric effects which occur when fatty acid synthesis is measured with³H₂O. The fatty acid composition in liver and mammary gland of dams and in liver and kidney of pups completely reflected dietary fatty acids. Columbinate made up ca. 20% of the total fatty acids in both tissues of the columbinic acid-fed mice and ca. 15% in the pup tissues. This suggests that columbinate is incorporated into milk lipids of dams and is easily absorbed by pups. The elevated ratios of 16/16∶1 and 18/18∶1 in liver and mammary gland of dams and liver and kidney of the pups from dams fed linoleate, linolenate and columbinate suggest that each of these polyunsaturated fatty acids in the diet can inhibit the activity of Δ9 desaturase.     

Biosynthesis of fatty acids from acetate in soybean suspension cultures

Suspension cultures of finely divided soybean cells established from callus were incubated with sodium [1(14)C] acetate for periods up to 86 hr. Lipids and fatty acids were analyzed for radioactivity in samples harvested at logarithmic time periods. Incorporation of acetate into cell lipid was directly proportional to the logarithm of time up to 32 hr, after an initial lag of 4-6 hr. Most of the lipid radioactivity was found in the phospholipid fraction, and all common soybean fatty acids became labeled within 6 hr. The order of labeling and distribution of radioactivity with time were essentially the same as in tissues from intact growing plants. These results support the concept of sequential desaturation of oleic acid in the cells. It was concluded that valid studies of the biosynthesis of common lipids in the soybean can be carried out for extended periods of time by use of undifferentiated cells in suspension cultures.     

Pulmonary surfactant lipid synthesis from ketone bodies,lactate and glucose in newborn rats

The contribution of acetoacetate (AcAc), β-hydroxybutyrate (βOHB), lactate and glucose to pulmonary surfactant lipid synthesis in three-to five-day-old rats was measured. Minced lung tissue was incubated with³H₂O and [3-¹⁴C]AcAc, [3-¹⁴C]βOHB, [U-¹⁴C]lactate or [U-¹⁴C]glucose, and the radioactivity incorporated into surfactant lipids was measured. When expressed as nmol of substrate incorporated/g lung tissue per four hr, lactate was incorporated more rapidly than other substrates into total surfactant lipids and phosphatidylcholine (PC). There was no difference in the rates of incorporation of lactate, AcAc or glucose into disaturated PC (DSPC). Substrates other than glucose were incorporated almost exclusively into fatty acids, whereas 60–80% of glucose incorporated into surfactant phospholipids was found in fatty acids, with the remaining in glyceride-glycerol. When expressed as nmol acetyl units incorporated/g lung tissue per four hr, the rates of AcAc, lactate and glucose incorporation into total surfactant fatty acids were comparable. Glucose incorporation into DSPC and PC was greater than that of AcAc and lactate. When glucose was the only exogenous substrate added to the incubation medium, it contributed 37% of total surfactant fatty acids synthesized de novo. In the presence of other substrates, the contribution of glucose to de novo fatty acid synthesis dropped to 14–20%. In the presence of unlabeled glucose,¹⁴C-labeled AcAc, lactate and βOHB contributed 52%, 40% and 19%, respectively, of the total fatty acids synthesized de novo. The rate of βOHB incorporation into surfactant lipids was only about 50% that of other substrates and was accompanied by low activity of β-hydroxybutyrate dehydrogenase measured for newborn lung. These results demonstrate that AcAc and lactate are important precursors for surfactant lipids in neonatal rat lung.     

Effect of reproductive states on lipid mobilization and linoleic acid metabolism in mammary glands

Effects of pregnancy and lactation on lipid metabolism in mouse mammary fat pads and nonmammary adipose tissues have been studied. In order to address the question whether the influence of hormonal milieu on lipid metabolism in mammary epithelial cells during pregnancy and lactation is the same as in fat cells, we have studied the mobilization of lipids and metabolism of fatty acids in the intact mammary glands, parenchyma-free mammary fat pads and in the perimetrial fat tissues of virgin, pregnant and lactating mice. Compared to parenchyma-free mammary fat pads, the perimetrial adipose tissues accumulated 5-fold higher levels of triglycerides during pregnancy. Mammary fat cells maintained overall lipid levels during pregnancy and lactation (16–20 μg/fat pad). In contrast, lactation depleted total lipid stores from 108 ± 5 to 24 ± 4.5 μg/fat pad in perimetrial fat pads. Results of comparative analysis of fatty acid composition of mammary fat pads, with and without epithelial tissue, from virgin and lactating mice showed stimulation of 18∶2ω6 metabolism leading to 130% increase in the ratio 20∶4ω6 to 18∶2ω6 in the epithelial compartment. Pregnancy and lactation resulted in the elevation of 20∶4ω6 levels probably due to a 4-fold increase in Δ5 desaturase activity and a decrease in oxidative degradation of 18∶2ω6. These results suggest that, unlike other adipose tissues, the metabolic pathways in mammary fat cells are not dedicated to sequestration and accumulation of dietary lipids during pregnancy. Lactation favors mammary epithelial cell-stimulated production of precursors of eicosanoids which are known to have agonist-like effect on mammary epithelial cells.