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Biomedical subjects

C S Fink

Publications and source records attributed to C S Fink.

At least 37 records · Page 2Linked to original sources

In vitro [U-14C]glucose utilization by tissues of weanling rats with lateral hypothalamic area lesions one month after lesion production.

The role of the lateral hypothalamic area (LHA) in intermediary metabolism was investigated by quantitation of [U-14C]glucose oxidation to 14CO2 and 14C incorporation into the glycogen and lipid fraction of the liver, epididymal fat pad, and diaphragm. Weanling male Sprague-Dawley rats received bilateral electrolytic lesions in the LHA (LHAL rats). Sham operated rats were either fed ad libitum (CON-ADLIB) or pair-gained to the LHAL rats (CON-PG). The experiment was terminated 1 month after lesion production. LHAL rats were significantly (SIG) lighter and shorter and ate less than CON-ADLIB; LHAL rats were also SIG shorter than CON-PG, pointing to a food intake-independent lesion effect. Both LHAL and CON-PG rats had SIG less percent carcass fat than CON-ADLIB, but there was no SIG difference between LHAL and CON-PG rats. Also, LHAL rats had a SIG higher percentage of carcass protein than both CON-ADLIB and CON-PG. Furthermore, LHAL rats incorporated SIG less glucose into liver glycogen than CON-ADLIB but SIG more into CON-PG, whereas CON-PG rats incorporated SIG less into liver glycogen than CON-ADLIB, again suggesting a food intake-independent effect. There was no difference among the groups in glucose oxidation and incorporation into lipids and glycogen in both diaphragm and epididymal fat pads and liver total lipid. However, livers of CON-PG metabolized SIG more [U-14C]glucose to CO2 than did livers of CON-ADLIB, suggesting a food intake-dependent effect. There was no difference between LHAL and CON-PG rats in this parameter.(ABSTRACT TRUNCATED AT 250 WORDS)

Adipose Tissue↗

In vitro activation and inhibition of rat colonic phospholipase D by fatty acids.

The present study was designed to examine the effect of long-chain fatty acids on phospholipase D (PLD) of the proximal colon mucosa of the rat. These fatty acids are present in the colon, in close contact with the membranes which contain the enzyme. The results indicate that unsaturated fatty acids activate the enzyme. At 4 mM, trienes are more potent activators as compared to 18:2 and 18:1. The number of double bonds and the configuration around the double bond were found to be important factors in activation of the enzyme. The metabolism of polyunsaturated fatty acids may be involved in the activation since the presence of antioxidants and cyclooxygenase inhibitors were found to influence the activation of PLD by polyunsaturated fatty acids.

Animals↗

Influence of dietary fat and feeding period on phosphoinositide metabolism in rat colonocytes.

The objective of the present study was to examine the effect of dietary fat content on phosphoinositide (PI) metabolism, fatty acid composition in colonocytes, and colonic luminal content of bile acids (BA) and free fatty acids (FFA) in rats. Male Sprague-Dawley rats weighing approximately 166 g were fed to semipurified diet containing 3% or 21.5% beef fat and 2% corn oil. The nonfat ingredients were adjusted to correct for differences in food consumption of these diets. Animals were fed these diets ad libitum for one or four weeks. The isolated colonocytes had a viability of 88.9% in all groups. PI metabolism was examined in the absence (basal) or presence of agonists, 2 mM deoxycholic acid or 10 microM A23187. Dietary fat concentration had no effect on PI metabolism, but the length of feeding had a significant effect on basal and stimulated PI metabolism. Colonocytes of animals fed the diets for four weeks were less sensitive to stimulation of PI cycle by agonists than those of animals fed for one week. Colonocyte fatty acid composition was influenced by dietary fat and feeding period. Only the relative percentage of 20:3(n-6) was significantly lower in rats fed the high-fat diet for one week; 18:0 was lower and 18:3(n-6) was higher in colonocytes of animals fed the diets for one week than in those fed for four weeks. Several colonic fatty acids, namely, 16:0, 20:3(n-6), and 22:5(n-6), also exhibited diet-by-feeding period interaction. Intracolonic luminal contents from rats fed the high-fat diet contained elevated concentrations of BA and FFA (44% and 62%, respectively). It was concluded that despite the effects of dietary fat concentration on increased colonic BA and FFA and on altered membrane fatty acid composition, dietary fat had no effect on PI metabolism in colonocytes under the conditions in the present experiment. A difference in components between the purified diet and the commercial rat chow and/or an aging effect of the rats may alter the PI cycle of colonocytes.

Animals↗

Differential response of glucose utilization by three rat muscle tissues to dietary fatty acid composition.

Rats were fed a semisynthetic diet containing 14% of either beef fat, safflower oil, or menhaden oil plus 2% corn oil for 7 weeks, and three tissues, diaphragm, heart, and skeletal muscle, were examined for fatty acid composition in their phospholipids and triglycerides. In addition, the lipid concentrations in these tissues were examined. The in vitro oxidation and incorporation of glucose into lipids of these tissues were also examined. Skeletal muscle showed the greatest change in phospholipid composition with diet. All tissues were responsive to changes in diet in regards to the triglyceride fraction. Dietary alteration of tissue phospholipid composition did not alter lipid concentration in these tissues. However, in diaphragm tissue, rats fed the beef fat diet had lower phospholipid and higher triglyceride synthesis compared with those fed either menhaden oil or safflower oil. In addition, triglyceride synthesis was higher in the diaphragm of animals fed the menhaden oil diet as compared with the safflower oil diet. Therefore, dietary fatty acid composition may play a role in the triglyceride and phospholipid metabolism of rat diaphragm.

Animals↗

Alteration of membrane fatty acid composition and inositol phosphate metabolism in HT-29 human colon cancer cells.

The present study was designed to investigate the role of membrane fatty acid (FA) composition on inositol phosphate (InsP) release by a human colon tumor cell line. Cells were supplemented for five days in culture with 0, 10, 30, or 100 microM sodium stearate (18:0), linoleate [18:2(omega-6)], or linolineate [18:3(omega-3)]. These FAs were supplied as a complex with FA-free bovine serum albumin. InsP release was examined in these cells with or without stimulation with deoxycholic acid (DCA) after they were labeled with [3H]myoinositol. FA enrichment was found to influence inositol incorporation into membrane lipids. Although 18:0 had no effect, 18:2(omega-6) decreased the incorporation. On the other hand, 18:3(omega-3) increased the incorporation of inositol compared with the cells supplemented with the other FAs, but they were not different from control. Basal release of total InsP was elevated only with supplementation of 10 and 30 microM 18:3(omega-3). FA supplementation with 18:0 at 30 microM and 18:2 at 30 and 100 microM resulted in downregulation of bsal release of InsP. Enrichment of HT-29 cell membranes with polyunsaturated FAs resulted in a significant increase in stimulated release of InsP, but this was not seen with saturated FA supplementation. At 10 microM supplementation, 18:2 had the greatest effect on stimulated InsP release. This effect of 18:2 disappeared at 30 microM. However, the increase in the stimulated InsP release caused by 18:3 occurred at 10 and 30 microM. DCA-stimulated release of InsP was not downregulated by any FA supplementation. This study showed that enrichment of the membranes with polyunsaturated FAs increases the response of the phosphatidylinositol cycle to DCA stimulation. In addition, enrichment with 18:3(omega-3) increases the basal turnover of InsP. It is concluded that alteration of membrane FAs has a profound effect on the phosphatidylinositol cycle.

Colonic Neoplasms↗

Effect of dietary fat on glomerular lipid composition and angiotensin II receptors.

Nutritional factors are major etiologic determinants which may affect the incidence or severity of hypertension. Since angiotensin II (Ang II) has a role in some forms of hypertension, the influence of dietary lipid composition on glomerular lipid content and on Ang II receptor parameters was determined. Three groups of rats were fed diets containing 14% by weight of 1) beef fat (saturated fatty acid rich), 2) safflower oil (n-6 fatty acid rich), or 3) fish oil (n-3 fatty acid rich), each supplemented with 2% corn oil. A fourth group of rats was fed rat chow which contained 6.5% fat. At the end of 7 weeks, the rats were sacrificed, the kidneys removed, and renal glomeruli isolated using a sieving technique. Feeding diets which varied in the quantity and composition of fatty acids altered glomerular fatty acid composition of n-6 and n-3 fatty acids and the unsaturation indices of glomeruli. The cholesterol content, but not the phospholipid content, was decreased in glomeruli of animals fed the high fat diets. This resulted in a lower cholesterol/phospholipid ratio, indicative of an increase in membrane fluidity. Glomerular binding was determined using 125I-Ang II in a radioreceptor assay. Binding was proportional to glomerular protein and was time dependent. Receptor affinity (Ka) and number were determined by Scatchard analysis of binding inhibition data. Glomerular Ka was significantly lower for animals fed the rat chow as compared to the semisynthetic diets. There was no significant difference in the concentration of Ang II receptors in glomeruli of animals fed the different diets. The data indicate that the induced alteration in glomerular lipid composition is associated with alteration in the binding affinity of glomerular Ang II receptors.

Animals↗

Dietary fat and phospholipase A2 activity of Sprague-Dawley rat large intestine.

The present studies were conducted to examine the effect of dietary lipid content and composition [(n-6) vs. (n-3) fatty acids] on the activity of mucosal phospholipase (PL)A2 of the large intestinal tract of rats. Three segments of the large intestinal tract were examined: cecum, proximal colon and distal colon. Weanling male Sprague-Dawley rats were fed diets containing either 5% (LS) or 16% safflower (HS) or 14% menhaden oil plus 2% safflower oil (HM) for 3 wk with the oil replacing starch in the HS and HM diets on a weight basis. The lipid extracts of microsomal fractions from mucosal scrapings were examined for phospholipid and cholesterol content and fatty acid composition. Phospholipase A2 was assayed using a fluorescent substrate. Rats fed the high fat diets had lower PLA2 specific activities. The (n-3) or (n-6) fatty acid enrichment of the membranes had no effect of the activity of the enzyme. The activity of the enzyme decreased aborally from the cecum to the distal colon; the proximal colon had an intermediate specific activity.

Animal Nutritional Physiological Phenomena↗

Fat absorption in premature infants: medium-chain triglycerides and long-chain triglycerides are absorbed from formula at similar rates.

Fat absorption from two different premature infant formulas and one full-term formula containing three different fat blends was investigated in two groups of premature infants. The first group of nine infants (gestational age, 29.1 +/- 0.88 weeks; postnatal age, 3.13 +/- 0.71 weeks) was fed alternately for 1 week each SMA preterm formula containing either high levels (50%) of medium-chain triglycerides (MCT) (6:0, 8:0, and 10:0) or high levels (86%) of long-chain triglycerides (LCT) (greater than or equal to C12). Except for fat blends, the formulas were otherwise identical. The second group of 11 infants (gestational age, 30.5 +/- 0.77 weeks, studied at a postnatal age of 4.33 +/- 0.91 weeks) was fed for 1 week a full-term infant formula, S-26, containing 98% LCT. Fat absorption (studied during a 3-day fat balance period) was similar irrespective of fat blend: 89.08 +/- 2.37% during feeding of preterm SMA, 50% MCT; 87.0 +/- 3.81% during feeding of preterm SMA, 86% LCT; and 83.00 +/- 2.89% during feeding of S-26, 98% LCT. Weight gain (grams per day) and increase in length (centimeters per day) were 23.2 +/- 1.7, 21.20 +/- 1.7, and 14.28 +/- 2.9, and 0.17 +/- 0.06, 0.16 +/- 0.04, and 0.22 +/- 0.07 during feeding of the three fat blends, respectively. Lipase activity levels in fasting gastric aspirates were higher during feeding of the LCT than the MCT formula. The possible stimulation of gastric lipase secretion secondary to long-chain fatty acid stimulation of cholecystokinin secretion might be related to the efficient digestion of formula fat, irrespective of triglyceride-fatty acid chain length.(ABSTRACT TRUNCATED AT 250 WORDS)

Dietary Fats↗

Effect of excess dietary calcium on colon mucosal membranes and fecal lipids.

The objective of the present studies was to examine the biochemical alterations in colon apical membranes upon feeding excess calcium to animals on a saturated fatty acid-rich diet. It has been suggested recently that excess dietary calcium may offer a protection to colon membranes as judged by histologic examination. Sprague-Dawley weanling male rats were fed a semisynthetic diet containing 14% beef fat plus 2% corn oil and either the calcium requirement or excess calcium in the form of calcium carbonate. Animals were fed the diets for 4 weeks. Feces were collected in the last 3 days. The results indicate that excess dietary calcium resulted in alteration in the density of 4 protein bands of colon apical membranes upon examination on SDS-gel electrophoresis. These bands contain 20% of membrane proteins. The diet had no effect on either the lipid content or fatty acid composition of the membranes. Excess dietary calcium resulted in a 54% reduction in fecal water bile acids and a 44% reduction in fecal water free fatty acids. The reduction in fecal water lipids was due to alterations in the solubility of these lipids. This was not mediated through alterations in the pH of fecal water. The observed alterations in protein patterns of these membranes may be due to either the reduction of fecal water bile acids and free fatty acids or may be a direct effect of dietary calcium on membrane proteins.

Animals↗

Effect of dietary fat on the lipid composition and utilization of short-chain fatty acids by rat colonocytes.

The objective of the present studies was to examine the effect of dietary fat on the lipid composition of rat colonocytes and their utilization of short-chain fatty acids (SCFA). Rats were fed 14% beef fat, fish oil or safflower oil plus 2% corn oil in a semi-synthetic base diet for 4 wk. Colonocytes were isolated and their lipid composition was examined. Feeding beef fat and fish oil resulted in an increase in monounsaturated fatty acids and a reduction in omega-6 fatty acids. Feeding fish oil resulted in an enrichment with omega-3 fatty acids. There was no dietary influence on the amount of either cholesterol or phospholipids of colonocytes. Fish oil feeding resulted in significant increase in colonocyte free fatty acids (FFA) as compared to other diets. Dietary fat was found to have no effect on SCFA utilization by colonocytes. Colonocytes were found to utilize SCFA in the order of butyrate greater than or equal to acetate greater than or equal to propionate. The presence of acetate and propionate in the medium had no effect on the rate of butyrate utilization.

Animals↗

Failure to demonstrate an effect of dietary fatty acid composition on body weight, body composition and parameters of lipid metabolism in mature rats.

The objectives of the present study were to examine the effects of dietary fatty acid composition on body composition and on several parameters of lipid metabolism in adipose tissue and to assess the ease of weight loss with restricted dietary intake. Rats were fed diets containing 14% fish oil, safflower oil or beef fat plus 2% corn oil. These diets were fed ad libitum for 4 wk (first phase). A number of the animals from each group were killed, and the others were fed 50% of their first-phase food intake for an additional 4 wk (second phase). The diets used in the second phase contained 3% of the above fats plus 2% corn oil. Food consumption, food efficiency and body weights were monitored. After the rats were killed, fatty acid composition of epididymal fat pads was examined. In addition, in vivo lipolysis and in vitro lipogenesis in epididymal fat pads were examined. The results indicate that dietary fatty acid composition had no effect on body weight, food consumption, in vivo lipolysis and in vitro lipogenesis in epididymal fat pads. In addition, although dietary fatty acid manipulation resulted in alteration in adipose tissue fatty acid composition, it had no effect on the rate of weight loss, body composition, in vivo lipolysis and in vitro lipogenesis in epididymal fat pads. It is concluded that dietary fatty acid composition does not play a role in body composition and in lipid metabolism in adipose tissue of mature rats.

Animals↗

Secretion of human gastric lipase from dispersed gastric glands.

The presence of a triacylglycerol lipase in human gastric juice was described in previous studies. Its source and role in intragastric lipolysis was, however, uncertain. Our study presents definitive evidence for gastric origin of a lipase and its release by secretagogues. Both carbachol and cholecystokinin-8 stimulate release of this enzyme for dispersed human gastric glands. While the two secretagogues had similar efficacies, with nearly a 3-fold stimulation over basal rates, cholecystokinin-8 was about four orders of magnitude more potent in releasing lipolytic activity than carbachol (maximum stimulation at concentrations of 1 X 10(-9) and 1 X 10(-5) M, respectively). Lipolytic activity measured against triolein (18:1), tricaprylin (8:0) and tributyrin (4:0) emulsions was 1.18 +/- 0.12, 4.48 +/- 0.64, and 12.17 +/- 0.88 units (1 unit = 1 mumol free fatty acid released/min per mg protein), respectively. Characterization of the pH optimum for each substrate showed maximum lipolysis at 4.5 for tributyrin, and at 5.5 for tricaprylin and triolein. These results indicate that a lipase which hydrolyzes long-, medium- and short-chain triacylglycerols is secreted by human gastric mucosa. At pH 6.0, the pH of the duodenum, there is appreciable lipolytic activity in the presence of bile salts. This suggests that gastric lipase, in addition to initiating lipolysis in the stomach, might also aid in the digestion of lipids in the duodenum. It remains to be determined whether gastric lipase is distinct from lingual lipase, or is the same enzyme secreted by the lingual serous glands and the gastric mucosa.

Bile Acids and Salts↗

Gastric lipolysis of milk lipids in suckling rats.

Fatty acid composition of the major lipid classes in stomach contents of suckling rats at 1, 5, 10, 17 and 20 days of lactation was compared to that of milk lipids. In milk, 98% of fatty acids were in triacylglycerols at all lactation times. Medium-chain fatty acid concentrations increased from 8% in colostrum to 26% at day 5. Fatty acid composition of stomach acylglycerols at all lactation times was different from that of milk triacylglycerols, containing less medium-chain fatty acids, 8:0 and 10:0. This preferential hydrolysis was also shown by higher concentrations of medium-chain fatty acids in the free fatty acid fraction. The lipolysis of medium-chain fatty acids from triacylglycerols resulted in the appearance of di- and monoacylglycerols with 50-100% higher amounts of 14:0 and 16:0. The similar fatty acid composition of products suggests that considerable lipolysis occurred in stomachs of suckling rats even at 1 day of age. Although there was a 10-fold increase in milk consumption, the extent of lipolysis was similar throughout the suckling period because of a parallel rise in lingual lipase levels.

Animal Population Groups↗

Lipase secretion from dispersed rabbit gastric glands.

We have measured gastric lipase activity in dispersed glands of rabbit stomach by quantitating the hydrolysis of tri[3H]olein. Lipase activity in isolated gastric glands was 200-400 nmol [3H]oleic acid released per milligram dry weight per minute. The percentage of lipase activity released during incubation for 30 min at 37 degrees C under basal conditions was 1.5-4.5%. Lipase release was stimulated by secretagogues: 10 nM cholecystokinin octapeptide (CCK-8) and 100 microM carbachol led to four- to sixfold and two- to threefold higher enzyme secretion, respectively, while histamine had no effect. Carbonyl cyanide m-chlorophenylhydrazone (30 microM) completely inhibited the CCK-8-induced lipase release, indicating that lipase secretion is dependent on mitochondrial oxidative energy; dibutyryl cGMP (1 mM) inhibited 1 nM CCK-8-stimulated but not 100 microM carbachol-stimulated secretion; atropine (1 microM) had the opposite effect. These studies suggest that secretion of lipase from isolated gastric glands is stimulated by at least two receptor mechanisms. These studies show that a lipase that hydrolyzes long-chain triglyceride is secreted by rabbit stomach mucosa and that the secretion of gastric lipase is stimulated by two different receptor mechanisms.

Animals↗

Fat digestion in the stomach: stability of lingual lipase in the gastric environment.

Digestion of dietary fat starts in the stomach, where lingual lipase hydrolyzes triglycerides to free fatty acids and partial glycerides at pH 3.0-6.0. Lingual lipase is secreted continuously from lingual serous glands and accumulates in the stomach between meals, when gastric pH is less than 3.0. We have, therefore, examined the resistance of lingual lipase to low pH and its possible protection by dietary components present in the stomach contents. Partially purified rat lingual lipase (7-15 micrograms enzyme protein) was preincubated at 37 degrees C for 10-60 min at pH 1.0-6.0 before incubation for assay of lipolytic activity, hydrolysis of tri-[3H]olein at pH 5.4. The data show that partially purified rat lingual lipase preparations are stable at 37 degrees C in the pH range of 2.5-6.0. Enzyme activity, however, is rapidly and irreversibly lost during preincubation at pH 1.0-2.4 for 10-30 min. Protein (gelatin 1% or albumin 1% or 2.5%) cannot prevent the inactivation of lingual lipase at low pH. The large molecular species (molecular weight greater than 500,000) of lingual lipase (thought to be an aggregate of enzyme with lipids) is slightly more resistant to inactivation than the 46,000 dalton preparation, suggesting that lipids might protect the enzyme from inactivation. Indeed, about 60% of the initial lipase activity is preserved during incubation at pH 2.0 in the presence of 50 mM lecithin or 10 mM triolein. The data indicate that triglycerides which are hydrolyzed by this enzyme as well as phospholipids that are not hydrolyzed can prevent the inactivation of the enzyme.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗

Peripheral serum corticosteroid concentrations in relation to the rat adrenal cortical circadian rhythm in androgen-induced hypertension.

Adrenal function was assessed in control rats and in rats treated for 2 and 4 weeks with 17 alpha-methylandrostenediol (MAD; 17 alpha-methyl-5-androstene-3 beta-diol), a synthetic androgen known to produce hypertensive cardiovascular disease. In both groups and at both time periods, a circadian rhythm of blood corticosteroid concentrations was observed. The high point for serum corticosterone (B), 18-hydroxy-11-deoxycorticosterone (18-hydroxy-DOC), and 11-deoxycorticosterone (DOC) concentrations occurred at the beginning of the dark period (1800 hours), and the low point occurred at the onset of the light period (0600 hours). Serum concentrations of DOC were always found to be higher in MAD-treated rats as compared with controls. The serum concentrations of B and 18-hydroxy-DOC were lower than control values at 1800 hours but higher than control concentrations at 0600 hours. The in vitro 11 beta- and 18-hydroxylation of DOC was markedly reduced with MAD treatment. In contrast, cholesterol side-chain cleavage activity was higher in animals treated with MAD. These in vitro findings correlated with spectral studies that showed a decreased binding of DOC to cytochrome P450(11) beta and increased binding of cholesterol to cytochrome P450scc. These studies suggest that MAD treatment selectively decreases 11 beta- and 18-hydroxylation in adrenal mitochondria, and this results in an increased serum concentration of DOC, a hypertensinogenic steroid. This effect of MAD on peripheral serum DOC concentration is most readily observed in quiescent animals at the high point of the circadian rhythm.

18-Hydroxycorticosterone↗

Dietary phytosterol inhibits the growth and metastasis of MDA-MB-231 human breast cancer cells grown in SCID mice.

The objective of the present study was to investigate the effect of dietary phytosterols on the growth and metastasis of the human breast cancer MDA-MB-231 cell line xenografted in SCID mice. Two groups of animals were fed AIN-93G diet supplemented with 0.2% cholic acid and 2% sterol (cholesterol or phytosterol mixture) for 15 days before inoculation of the tumor into the right inguinal mammary fat pad. Tumor growth and food consumption were recorded weekly throughout the 8 weeks of the experiment. At the end of the experiment, the animals fed phytosterol had a 40% lower serum cholesterol and 20 and 30 fold higher serum beta-sitosterol and campesterol, respectively as compared to those fed cholesterol. There was no difference between the two groups in body weight and food consumption. However, the tumor size in animals fed phytosterols was 33% smaller (P < 0.03) and had 20% fewer metastases to lymph nodes and lungs than the cholesterol group. At termination, the tumor weight of the animals fed the phytosterol diet was also less (P < 0.07) than that of the cholesterol group. It is concluded that dietary phytosterols retard the growth and spread of breast cancer cells.

Animals↗